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pubmed23n0125_13677

A thin-gel isoelectric focusing method for quantitation of protein S-thiolation.

A thin-gel isoelectric focusing method has been developed for analysis of protein S-thiolation (formation of mixed disulfides with low molecular weight thiols). The method is rapid and it can be used with 3 to 5 micrograms of a pure protein, or 15 to 20 micrograms of tissue extract protein. It is possible to detect a modification of the protein sulfhydryl by either charged or uncharged thiols, and to determine the quantity of different S-thiolated protein species in a modified sample. The method was used to quantitate the amount of S-thiolation of phosphorylase b in a reaction with oxidized glutathione that produced four S-thiolated forms of the enzyme. The method was also used to detect S-thiolation of two proteins in a cardiac tissue extract treated with diamide. One of the protein bands was shown to be S-thiolated with both cysteine and glutathione, while the other band was S-thiolated only with glutathione.

A thin-gel isoelectric focusing method for quantitation of protein S-thiolation. A thin-gel isoelectric focusing method has been developed for analysis of protein S-thiolation (formation of mixed disulfides with low molecular weight thiols). The method is rapid and it can be used with 3 to 5 micrograms of a pure protein, or 15 to 20 micrograms of tissue extract protein. It is possible to detect a modification of the protein sulfhydryl by either charged or uncharged thiols, and to determine the quantity of different S-thiolated protein species in a modified sample. The method was used to quantitate the amount of S-thiolation of phosphorylase b in a reaction with oxidized glutathione that produced four S-thiolated forms of the enzyme. The method was also used to detect S-thiolation of two proteins in a cardiac tissue extract treated with diamide. One of the protein bands was shown to be S-thiolated with both cysteine and glutathione, while the other band was S-thiolated only with glutathione.

pubmed23n0125_13678

Comparison of electron and chemical ionization mass spectrometry of sialic acids.

Sialic acids were analyzed as per-O-trimethylsilylated compounds by gas-liquid chromatography/mass spectrometry either on electron or chemical ionization by isobutane. Electron ionization mass spectra of these derivatives are very similar to those of the corresponding methyl esters described earlier whereas chemical ionization mass spectra are characterized in the high mass range by loss of the C-2 and the C-4 substituents from the M + 1 ion. Together with other fragment ions the seven different sialic acids analyzed could be clearly identified.

Comparison of electron and chemical ionization mass spectrometry of sialic acids. Sialic acids were analyzed as per-O-trimethylsilylated compounds by gas-liquid chromatography/mass spectrometry either on electron or chemical ionization by isobutane. Electron ionization mass spectra of these derivatives are very similar to those of the corresponding methyl esters described earlier whereas chemical ionization mass spectra are characterized in the high mass range by loss of the C-2 and the C-4 substituents from the M + 1 ion. Together with other fragment ions the seven different sialic acids analyzed could be clearly identified.

pubmed23n0125_13679

Preparation of rat liver plasma membrane with respect to glucagon-sensitive adenylate cyclase.

Further modification of Neville's method of preparation of rat liver plasma membrane has been made in order to study glucagon-sensitive adenylate cyclase. This modified method introduces two additional steps to the Neville procedures. One involves saving an intermediate layer above the pellet following 1500g centrifugation. The other adds a centrifugation step at 20,000g. The improved method increases the yield of membrane protein by 5-fold and increases forskolin, glucagon, and 5'-guanyl imidodiphosphate stimulated activity of adenylate cyclase by 2.8-fold. A 13-fold increase in the yield of total adenylate cyclase activity above the current method was obtained. The membrane adenylate cyclase and its hormone sensitivity was stable in liquid nitrogen for at least 8 months. This modified method appears to be useful in preparing a better yield and quality of rat liver plasma membrane from given starting hepatic tissue for studies of glucagon-sensitive adenylate cyclase.

Preparation of rat liver plasma membrane with respect to glucagon-sensitive adenylate cyclase. Further modification of Neville's method of preparation of rat liver plasma membrane has been made in order to study glucagon-sensitive adenylate cyclase. This modified method introduces two additional steps to the Neville procedures. One involves saving an intermediate layer above the pellet following 1500g centrifugation. The other adds a centrifugation step at 20,000g. The improved method increases the yield of membrane protein by 5-fold and increases forskolin, glucagon, and 5'-guanyl imidodiphosphate stimulated activity of adenylate cyclase by 2.8-fold. A 13-fold increase in the yield of total adenylate cyclase activity above the current method was obtained. The membrane adenylate cyclase and its hormone sensitivity was stable in liquid nitrogen for at least 8 months. This modified method appears to be useful in preparing a better yield and quality of rat liver plasma membrane from given starting hepatic tissue for studies of glucagon-sensitive adenylate cyclase.

pubmed23n0125_13680

Isolation of the zona pellucida and purification of its glycoprotein families from pig oocytes.

The isolation of the porcine zona pellucida, the glycoprotein envelope surrounding the mammalian oocyte, and the purification of its glycoprotein families is described. The zona pellucida was prepared from oocytes isolated from pig ovaries using a razor blade device and sieving procedures with Teflon or nylon screens. In 6-7 man h, the zona pellucida from 5 X 10(5) oocytes was obtained yielding 12 mg protein and 2.2 mg carbohydrate. The absorptivity of heat solubilized and filtered zona pellucida was A1%280 nm, 1 cm = 10.8. The four glycoprotein families of the zona pellucida were purified by two-dimensional polyacrylamide gel electrophoresis and electrophoretic elution. The electrophoretic purity of these families was greater than 90%. The protein and carbohydrate content and the amino acid and monosaccharide compositions of each of the glycoprotein families were determined.

Isolation of the zona pellucida and purification of its glycoprotein families from pig oocytes. The isolation of the porcine zona pellucida, the glycoprotein envelope surrounding the mammalian oocyte, and the purification of its glycoprotein families is described. The zona pellucida was prepared from oocytes isolated from pig ovaries using a razor blade device and sieving procedures with Teflon or nylon screens. In 6-7 man h, the zona pellucida from 5 X 10(5) oocytes was obtained yielding 12 mg protein and 2.2 mg carbohydrate. The absorptivity of heat solubilized and filtered zona pellucida was A1%280 nm, 1 cm = 10.8. The four glycoprotein families of the zona pellucida were purified by two-dimensional polyacrylamide gel electrophoresis and electrophoretic elution. The electrophoretic purity of these families was greater than 90%. The protein and carbohydrate content and the amino acid and monosaccharide compositions of each of the glycoprotein families were determined.

pubmed23n0125_13681

A procedure for the immunoblotting of proteins separated on isoelectric focusing gels.

A method has been devised for performing Western blot assays on proteins resolved by isoelectric focusing. Electrophoretic transfer of proteins directly from isoelectric focusing (IEF) tube gels to nitrocellulose sheets allowed their immunoassay without conventional second dimension SDS gel electrophoresis. The same method can also be used for IEF slab gels. For the immunostaining of nonmuscle actin isoforms in extracts of cultured cells, the resolution of this technique was much improved over that of Western blots of two-dimensional gels.

A procedure for the immunoblotting of proteins separated on isoelectric focusing gels. A method has been devised for performing Western blot assays on proteins resolved by isoelectric focusing. Electrophoretic transfer of proteins directly from isoelectric focusing (IEF) tube gels to nitrocellulose sheets allowed their immunoassay without conventional second dimension SDS gel electrophoresis. The same method can also be used for IEF slab gels. For the immunostaining of nonmuscle actin isoforms in extracts of cultured cells, the resolution of this technique was much improved over that of Western blots of two-dimensional gels.

pubmed23n0125_13682

Rapid sulfopropyl-disk chromatographic purification of bovine and human thrombin.

Thrombin, from either a crude commercial preparation (bovine) or a prothrombin activation mixture (human), was purified by sulfopropyl-disk chromatography to homogeneity in a rapid and convenient single-step procedure. The yield of both proteinases was greater than 85%. Purified bovine and human thrombin had sp act of 2500 and 3000 "NIH" units/mg, respectively. Human thrombin was more reactive than bovine thrombin with these active site-directed probes: phenylmethylsulfonyl fluoride, dansyl-Glu-Gly-Arg-chloromethylketone, and human heparin cofactor II. The sulfopropyl-disk chromatographic method is a useful and rapid technique to prepare milligram quantities of highly purified bovine and human thrombin.

Rapid sulfopropyl-disk chromatographic purification of bovine and human thrombin. Thrombin, from either a crude commercial preparation (bovine) or a prothrombin activation mixture (human), was purified by sulfopropyl-disk chromatography to homogeneity in a rapid and convenient single-step procedure. The yield of both proteinases was greater than 85%. Purified bovine and human thrombin had sp act of 2500 and 3000 "NIH" units/mg, respectively. Human thrombin was more reactive than bovine thrombin with these active site-directed probes: phenylmethylsulfonyl fluoride, dansyl-Glu-Gly-Arg-chloromethylketone, and human heparin cofactor II. The sulfopropyl-disk chromatographic method is a useful and rapid technique to prepare milligram quantities of highly purified bovine and human thrombin.

pubmed23n0125_13683

Hydrophobicity modulation as a tool for the purification of histidyl peptides.

A methodology is described for purification of histidyl peptides based on the changes in hydrophobicity induced by the specific and reversible modification of histidine residues by ethoxyformic anhydride. The mixture of modified peptides is subjected to HPLC; peptides containing modified histidines are detected at 242 nm, recovered, deethoxyformylated, and rechromatographed under the same conditions, then being detected at 220 nm. This procedure allows their isolation free from contaminants.

Hydrophobicity modulation as a tool for the purification of histidyl peptides. A methodology is described for purification of histidyl peptides based on the changes in hydrophobicity induced by the specific and reversible modification of histidine residues by ethoxyformic anhydride. The mixture of modified peptides is subjected to HPLC; peptides containing modified histidines are detected at 242 nm, recovered, deethoxyformylated, and rechromatographed under the same conditions, then being detected at 220 nm. This procedure allows their isolation free from contaminants.

pubmed23n0125_13684

Synthesis and NMR spectra of 13C-labeled coenzyme A esters.

The synthesis of coenzyme A thioesters of 13C-labeled acetate, propionate, succinate, and methyl malonate is described. The average yields were 94%. The 13C-NMR spectra were determined to provide a reference for the resonance positions of these metabolites. The synthesis of coenzyme thioesters of small-molecular-weight acids labeled with 13C has not been described previously, nor have the resonance positions been previously reported.

Synthesis and NMR spectra of 13C-labeled coenzyme A esters. The synthesis of coenzyme A thioesters of 13C-labeled acetate, propionate, succinate, and methyl malonate is described. The average yields were 94%. The 13C-NMR spectra were determined to provide a reference for the resonance positions of these metabolites. The synthesis of coenzyme thioesters of small-molecular-weight acids labeled with 13C has not been described previously, nor have the resonance positions been previously reported.

pubmed23n0125_13685

Binding pattern of ferritin-labeled lectins (RCAI and WGA) during neural tube closure in the bantam embryo.

Cell surface sugar residues in neurulating ectoderms of bantam chick embryos of stage 4-11 were examined using ferritin-labeled Ricinus communis (RCAI) and Wheat germ agglutinin (WGA). RCAI binding sites densely covered the apical surfaces of the basal plate cells during the neural plate stage (1,322.8 +/- 28.8 ferritin particles/micron 2). As neural tube formation advanced, the number of receptors decreased as a result of an increase in the extent of the sparsely covered regions. The decrease in receptors for WGA occurred in a similar manner but more rapidly. By the stage of development at which the opposite sides of the neural ridges meet at the dorsal midline, the receptors for WGA were reduced to about half. After this period, the two lectin receptors did not show significant changes. This result suggests that sugar residues or the sugar-chain skeleton on basal plate cells are altered during neurulation.

Binding pattern of ferritin-labeled lectins (RCAI and WGA) during neural tube closure in the bantam embryo. Cell surface sugar residues in neurulating ectoderms of bantam chick embryos of stage 4-11 were examined using ferritin-labeled Ricinus communis (RCAI) and Wheat germ agglutinin (WGA). RCAI binding sites densely covered the apical surfaces of the basal plate cells during the neural plate stage (1,322.8 +/- 28.8 ferritin particles/micron 2). As neural tube formation advanced, the number of receptors decreased as a result of an increase in the extent of the sparsely covered regions. The decrease in receptors for WGA occurred in a similar manner but more rapidly. By the stage of development at which the opposite sides of the neural ridges meet at the dorsal midline, the receptors for WGA were reduced to about half. After this period, the two lectin receptors did not show significant changes. This result suggests that sugar residues or the sugar-chain skeleton on basal plate cells are altered during neurulation.

pubmed23n0125_13686

Ultrastructural observations on nucleoli and related structures during human spermatogenesis.

The ultrastructural study of nucleoli and ribonucleoprotein-containing structures in human seminiferous tubules revealed that the nucleoli of spermatogonia, spermatocytes and Sertoli cells exhibited a tripartite structure consisting of: a fibrillar center, a compact granular portion, and a reticular portion containing both pars fibrosa and pars granulosa. The nucleoli of primary spermatocytes showed a developed reticular portion. At pachytene, the compact granular portion enlarged and lost its connection with the fibrillar center and the reticular portion which decreased in size. This suggests a nucleolar segregation similar to that of ovocytes in many species. Two similar developmental stages of nucleoli were observed in spermatogonia. In addition to nucleoli, there were other ribonucleoprotein-containing structures such as intranuclear closely-packed granules in Ap spermatogonia, coarse granules in the chromatin rarefaction zone of Ad spermatogonia, the nuage and Lubarsch crystals of spermatogonia, the chromatoid body of spermatids, the annulate lamellae of both spermatids and Sertoli cells, and many structures of the spermatid neck region.

Ultrastructural observations on nucleoli and related structures during human spermatogenesis. The ultrastructural study of nucleoli and ribonucleoprotein-containing structures in human seminiferous tubules revealed that the nucleoli of spermatogonia, spermatocytes and Sertoli cells exhibited a tripartite structure consisting of: a fibrillar center, a compact granular portion, and a reticular portion containing both pars fibrosa and pars granulosa. The nucleoli of primary spermatocytes showed a developed reticular portion. At pachytene, the compact granular portion enlarged and lost its connection with the fibrillar center and the reticular portion which decreased in size. This suggests a nucleolar segregation similar to that of ovocytes in many species. Two similar developmental stages of nucleoli were observed in spermatogonia. In addition to nucleoli, there were other ribonucleoprotein-containing structures such as intranuclear closely-packed granules in Ap spermatogonia, coarse granules in the chromatin rarefaction zone of Ad spermatogonia, the nuage and Lubarsch crystals of spermatogonia, the chromatoid body of spermatids, the annulate lamellae of both spermatids and Sertoli cells, and many structures of the spermatid neck region.

pubmed23n0125_13687

A quantitative electron microscopic study of the effect of glucocorticoids in vivo on the early postnatal differentiation of paraneuronal cells in the carotid body and the adrenal medulla of the rat.

The postnatal differentiation of carotid body chief cells and endocrine adrenal medullary cells was comparatively examined during ontogenesis and in rats which were treated with dexamethasone for 7 days after birth. Ultrastructure and innervation of carotid body chief cells are mature in neonates according to the functional requirements of chemoreception. By the end of the first postnatal week, only an increase in number of dense core vesicles can be noticed, the concentration of which then will reach the adult level. Under the effect of dexamethasone most of the heterochromatin is transformed into finely dispersed euchromatin within the nuclei of carotid body chief cells. In the cytoplasm, the Golgi apparatus becomes larger and the granular endoplasmic reticulum hypertrophic. The number of catecholamines storing dense core vesicles increases considerably. The innervation density remains constant. In contrast to the carotid body chief cells, the adrenal medullary cells have not reached their definitive maturity at the time of birth. Besides phenotypes of adrenaline-cells, noradrenaline-cells and small granules containing cells, pheochromoblasts and intermediary cells can be seen as well. Their cytoplasm is sparse, the concentration of dense core vesicles and the innervation density very low. After 8 days of postnatal ontogenesis, pheochromoblasts and intermediary cells are no longer present in the adrenal medulla. In adrenaline-cells and noradrenaline-cells, important processes of growth can be noticed, the cytoplasm has grown in extent, the number of dense core vesicles doubled and the innervation density of single cells triplicated. Only the few small granules containing cells remain small. Under the effect of dexamethasone also in the nuclei of chromaffin cells a transformation of heterochromatin into euchromatin occurs. The increase in number of dense core vesicles is relatively lower than in carotid body chief cells. The significant growth of innervation density during the first postnatal week was inhibited. Our observations suggest that dexamethasone stimulates the synthesis of catecholamines in adrenal medullary cells of newborn rats less pronouncedly than in carotid body chief cells. This could be attributed to the inhibited formation of synapses of growing chromaffin cells and to the in vivo active endocrine counter-regulation.

A quantitative electron microscopic study of the effect of glucocorticoids in vivo on the early postnatal differentiation of paraneuronal cells in the carotid body and the adrenal medulla of the rat. The postnatal differentiation of carotid body chief cells and endocrine adrenal medullary cells was comparatively examined during ontogenesis and in rats which were treated with dexamethasone for 7 days after birth. Ultrastructure and innervation of carotid body chief cells are mature in neonates according to the functional requirements of chemoreception. By the end of the first postnatal week, only an increase in number of dense core vesicles can be noticed, the concentration of which then will reach the adult level. Under the effect of dexamethasone most of the heterochromatin is transformed into finely dispersed euchromatin within the nuclei of carotid body chief cells. In the cytoplasm, the Golgi apparatus becomes larger and the granular endoplasmic reticulum hypertrophic. The number of catecholamines storing dense core vesicles increases considerably. The innervation density remains constant. In contrast to the carotid body chief cells, the adrenal medullary cells have not reached their definitive maturity at the time of birth. Besides phenotypes of adrenaline-cells, noradrenaline-cells and small granules containing cells, pheochromoblasts and intermediary cells can be seen as well. Their cytoplasm is sparse, the concentration of dense core vesicles and the innervation density very low. After 8 days of postnatal ontogenesis, pheochromoblasts and intermediary cells are no longer present in the adrenal medulla. In adrenaline-cells and noradrenaline-cells, important processes of growth can be noticed, the cytoplasm has grown in extent, the number of dense core vesicles doubled and the innervation density of single cells triplicated. Only the few small granules containing cells remain small. Under the effect of dexamethasone also in the nuclei of chromaffin cells a transformation of heterochromatin into euchromatin occurs. The increase in number of dense core vesicles is relatively lower than in carotid body chief cells. The significant growth of innervation density during the first postnatal week was inhibited. Our observations suggest that dexamethasone stimulates the synthesis of catecholamines in adrenal medullary cells of newborn rats less pronouncedly than in carotid body chief cells. This could be attributed to the inhibited formation of synapses of growing chromaffin cells and to the in vivo active endocrine counter-regulation.

pubmed23n0125_13688

Quantitative cellular changes during postnatal development of the rat dorsal lateral geniculate nucleus.

Quantitative changes in cell number during development of the dorsal lateral geniculate nucleus were determined using semithin serial sections of tissue obtained from 28 rats on postnatal day 0, 5, 8, 10, 20, 30, 90 or 165. Our results show three phases of postnatal development in the rat dorsal lateral geniculate nucleus: phase 1 from birth until eye opening, which occurs around the 12th day in these litters; phase 2 from eye opening through stabilization of neuron number on the 30th postnatal day, and phase 3 from that event until adulthood. During the first period increases in neuron number and in glial cell number are found accompanying a nearly seven-fold increase in dorsal lateral geniculate nucleus volume. Phase 2 includes a high incidence of neuronal cell death and a continuous increase in the number of glial cells. The third phase is characterized by a stabilization in the number of neurons, although the glial cell number continues to increase. Neuronal density decreases exponentially throughout the postnatal life of the rat, while the density of glial cells remains relatively stable over the period of study. The postnatal phenomenon of an initial increase in neuron number followed by a period of neuron death may be related to modulating and plastic functions which occur in the rat dorsal lateral geniculate nucleus before a stable neuronal population is achieved on the 30th postnatal day.

Quantitative cellular changes during postnatal development of the rat dorsal lateral geniculate nucleus. Quantitative changes in cell number during development of the dorsal lateral geniculate nucleus were determined using semithin serial sections of tissue obtained from 28 rats on postnatal day 0, 5, 8, 10, 20, 30, 90 or 165. Our results show three phases of postnatal development in the rat dorsal lateral geniculate nucleus: phase 1 from birth until eye opening, which occurs around the 12th day in these litters; phase 2 from eye opening through stabilization of neuron number on the 30th postnatal day, and phase 3 from that event until adulthood. During the first period increases in neuron number and in glial cell number are found accompanying a nearly seven-fold increase in dorsal lateral geniculate nucleus volume. Phase 2 includes a high incidence of neuronal cell death and a continuous increase in the number of glial cells. The third phase is characterized by a stabilization in the number of neurons, although the glial cell number continues to increase. Neuronal density decreases exponentially throughout the postnatal life of the rat, while the density of glial cells remains relatively stable over the period of study. The postnatal phenomenon of an initial increase in neuron number followed by a period of neuron death may be related to modulating and plastic functions which occur in the rat dorsal lateral geniculate nucleus before a stable neuronal population is achieved on the 30th postnatal day.

pubmed23n0125_13689

A study of lamellar organisation in juvenile and adult human bone.

The scanning electron microscope (SEM) has been used to study the three-dimensional organisation of collagen in slices of human rib and femur which were "etched" by chick osteoclasts, mechanically isolated and grown on their surfaces in vitro. Collagen organisation in the two bones showed a spectrum of appearances, ranging from lamellae of approximately equal thickness, but alternating fibre orientations, to an almost exclusive orientation of collagen apparently in a longitudinal direction. The rib contained a smaller component of transversely oriented collagen which may be related to a different functional loading. The thickness of circumferential lamellae was less than that of osteonal lamellae in the two adult ribs examined. Also, in the rib there was a trend towards increased average lamellar thickness with age in the range studied. This may be related to the fact that more of the lamellae in the rib cortex in children have been formed circumferentially. Correlation of results obtained with the SEM and the polarised light microscope (PLM) from the same substratum demonstrated that the latter grossly exaggerated the apparent component of collagen with a transverse orientation. This will always be true unless sections comparable with the lamellar thickness are used with the PLM.

A study of lamellar organisation in juvenile and adult human bone. The scanning electron microscope (SEM) has been used to study the three-dimensional organisation of collagen in slices of human rib and femur which were "etched" by chick osteoclasts, mechanically isolated and grown on their surfaces in vitro. Collagen organisation in the two bones showed a spectrum of appearances, ranging from lamellae of approximately equal thickness, but alternating fibre orientations, to an almost exclusive orientation of collagen apparently in a longitudinal direction. The rib contained a smaller component of transversely oriented collagen which may be related to a different functional loading. The thickness of circumferential lamellae was less than that of osteonal lamellae in the two adult ribs examined. Also, in the rib there was a trend towards increased average lamellar thickness with age in the range studied. This may be related to the fact that more of the lamellae in the rib cortex in children have been formed circumferentially. Correlation of results obtained with the SEM and the polarised light microscope (PLM) from the same substratum demonstrated that the latter grossly exaggerated the apparent component of collagen with a transverse orientation. This will always be true unless sections comparable with the lamellar thickness are used with the PLM.

pubmed23n0125_13690

Ontogenesis of the laminar structure in areas 17 and 18 of the human visual cortex. A quantitative study.

A quantitative morphological study of the pre- and postnatal development in the primary (area 17) and secondary (area 18) visual cortical regions was performed on 108 human brains. The neuropil proportion and thickness were measured with an image analyzer for the different cortical layers and the resulting data were approximated with logistic growth functions. The different layers show a marked heterochrony both within and between the areas. The neuropil proportion of layer I is the compartment to develop first in both areas. It has the lowest growth velocity, followed by layer VI and layers V, IV, III and II. This maturational sequence reflects the sequence of appearance of immature neurons during the migration period of neocortical ontogenesis. The development of the neuropil proportion is highly synchronized between areas 17 and 18 during the prenatal period, but in the first postnatal weeks, area 17 grows more quickly than area 18. Later on, this relation is reversed and area 18 reaches adult values of neuropil proportions about three months earlier than area 17. The growth in thickness of all layers is completed later than the growth in neuropil proportion. The growth in layer thickness is completed in Area 18 about two months earlier than in area 17, although area 18 has a greater cortical thickness. The results are compared with data on growth in volume, dendritic arborization and the development of visual function.

Ontogenesis of the laminar structure in areas 17 and 18 of the human visual cortex. A quantitative study. A quantitative morphological study of the pre- and postnatal development in the primary (area 17) and secondary (area 18) visual cortical regions was performed on 108 human brains. The neuropil proportion and thickness were measured with an image analyzer for the different cortical layers and the resulting data were approximated with logistic growth functions. The different layers show a marked heterochrony both within and between the areas. The neuropil proportion of layer I is the compartment to develop first in both areas. It has the lowest growth velocity, followed by layer VI and layers V, IV, III and II. This maturational sequence reflects the sequence of appearance of immature neurons during the migration period of neocortical ontogenesis. The development of the neuropil proportion is highly synchronized between areas 17 and 18 during the prenatal period, but in the first postnatal weeks, area 17 grows more quickly than area 18. Later on, this relation is reversed and area 18 reaches adult values of neuropil proportions about three months earlier than area 17. The growth in thickness of all layers is completed later than the growth in neuropil proportion. The growth in layer thickness is completed in Area 18 about two months earlier than in area 17, although area 18 has a greater cortical thickness. The results are compared with data on growth in volume, dendritic arborization and the development of visual function.

pubmed23n0125_13691

Development of the lateral amygdaloid nucleus in the human fetus: transient presence of discrete cytoarchitectonic units.

The cytoarchitectonic development of the lateral amygdaloid nucleus has been studied on Nissl-stained sections through brains of human fetuses ranging between 11 to 24 weeks of gestation. The first sign of cytoarchitectonic inhomogeneity of the lateral amygdaloid nucleus is the appearance of 2-3 ovoid cell clusters around 12 weeks of gestation. Between 12.5-16 weeks of gestation, the ventral part of the lateral amygdaloid nucleus contains 7-11 columnar cell clusters separated by "septa" of lower cell-packing density. These columnar clusters, stretching in the rostrocaudal direction, appear on cross-section as ovoid structures elongated in the ventrodorsal direction. In subsequent development (16-24 weeks of gestation) this distinct columnar appearance becomes less obvious, owing to the disappearance of "septa" along the dorsal edges of cellular clusters. This process begins first in the medial part of the columnar field. As a result, the cytoarchitectonic units gradually fuse into a homogeneous grey mass. However, the ventral part of the columnar field retains an undulated appearance throughout late gestation, showing multiple indentations as a sign of former cytoarchitectonic inhomogeneities. In conclusion, the fetal lateral amygdaloid nucleus contains a number of cytoarchitectonic "moduli" which could serve as a new parameter for an estimation of histogenetic maturity of the human amygdala. This transient cytoarchitectonic inhomogeneity could be a sign of the temporary predominance of one characteristic afferent-efferent system during a given developmental stage. Alternatively, it could reflect a clustered type of neurogenesis.

Development of the lateral amygdaloid nucleus in the human fetus: transient presence of discrete cytoarchitectonic units. The cytoarchitectonic development of the lateral amygdaloid nucleus has been studied on Nissl-stained sections through brains of human fetuses ranging between 11 to 24 weeks of gestation. The first sign of cytoarchitectonic inhomogeneity of the lateral amygdaloid nucleus is the appearance of 2-3 ovoid cell clusters around 12 weeks of gestation. Between 12.5-16 weeks of gestation, the ventral part of the lateral amygdaloid nucleus contains 7-11 columnar cell clusters separated by "septa" of lower cell-packing density. These columnar clusters, stretching in the rostrocaudal direction, appear on cross-section as ovoid structures elongated in the ventrodorsal direction. In subsequent development (16-24 weeks of gestation) this distinct columnar appearance becomes less obvious, owing to the disappearance of "septa" along the dorsal edges of cellular clusters. This process begins first in the medial part of the columnar field. As a result, the cytoarchitectonic units gradually fuse into a homogeneous grey mass. However, the ventral part of the columnar field retains an undulated appearance throughout late gestation, showing multiple indentations as a sign of former cytoarchitectonic inhomogeneities. In conclusion, the fetal lateral amygdaloid nucleus contains a number of cytoarchitectonic "moduli" which could serve as a new parameter for an estimation of histogenetic maturity of the human amygdala. This transient cytoarchitectonic inhomogeneity could be a sign of the temporary predominance of one characteristic afferent-efferent system during a given developmental stage. Alternatively, it could reflect a clustered type of neurogenesis.

pubmed23n0125_13692

The morphology of neurons in rat tectal transplants as revealed by Golgi-Cox impregnation.

The Golgi technique has been used to examine the morphology of neurons within tectal transplants. Embryonic tectal tissue was transplanted to the midbrain of newborn rats. Four to eight months later, host animals were decapitated under anaesthesia, the unfixed brains removed and processed by Golgi-Cox impregnation. In tectal grafts, different types of neuron were recognized on the basis of the size and shape of their somata and the morphology of their dendritic trees. Neuronal types found in transplants resembled cell classes found in normal rat superior colliculus (SC). Neurons characteristic of the superficial collicular layers such as marginal, ganglion type I, stellate and horizontal cells and multipolar cells typical of the deeper collicular layers were identified in the transplants. Compared with normal cells, grafted neurons often had smaller dendritic fields and fewer dendritic spines. No laminar organization was discernable in the grafts and there was commonly no preferential orientation of perikarya or dendrites. Small cells with similar dendritic morphology were sometimes found grouped together in patches within the graft neuropil. These patches resembled cytologically and histochemically distinct areas described in previous studies and may represent areas homologous to the superficial layers of normal SC.

The morphology of neurons in rat tectal transplants as revealed by Golgi-Cox impregnation. The Golgi technique has been used to examine the morphology of neurons within tectal transplants. Embryonic tectal tissue was transplanted to the midbrain of newborn rats. Four to eight months later, host animals were decapitated under anaesthesia, the unfixed brains removed and processed by Golgi-Cox impregnation. In tectal grafts, different types of neuron were recognized on the basis of the size and shape of their somata and the morphology of their dendritic trees. Neuronal types found in transplants resembled cell classes found in normal rat superior colliculus (SC). Neurons characteristic of the superficial collicular layers such as marginal, ganglion type I, stellate and horizontal cells and multipolar cells typical of the deeper collicular layers were identified in the transplants. Compared with normal cells, grafted neurons often had smaller dendritic fields and fewer dendritic spines. No laminar organization was discernable in the grafts and there was commonly no preferential orientation of perikarya or dendrites. Small cells with similar dendritic morphology were sometimes found grouped together in patches within the graft neuropil. These patches resembled cytologically and histochemically distinct areas described in previous studies and may represent areas homologous to the superficial layers of normal SC.

pubmed23n0125_13693

Development of the auditory hair cell surface in human fetuses. A scanning electron microscopy study.

Thirteen human fetal cochleas were investigated by scanning electron microscopy. Our observations concentrated on the hair cell surface. Ciliogenesis appeared to start during the 11th week of gestation on the inner hair cells (IHCs) and one week later on the outer hair cells (OHCs). The earliest stages of stereociliary development were similar on both types of cell and were characterized by the presence of round bundles of cilia arising from the surrounding microvilli. A three-dimensional V-shaped arrangement suddenly appeared, accompanied by the disappearance of short cilia on the internal side. Between the 20th and the 22nd weeks of gestation, both types of hair cell had an adult stereociliary pattern, i.e. a rectilinear arrangement on IHCs and W-shaped on OHCs. However, there were signs of immaturity, such as a disarray of OHCs and the presence of the kinocilium, suggesting that the surface of the auditory hair cells achieves its maturation during the last trimester of pregnancy.

Development of the auditory hair cell surface in human fetuses. A scanning electron microscopy study. Thirteen human fetal cochleas were investigated by scanning electron microscopy. Our observations concentrated on the hair cell surface. Ciliogenesis appeared to start during the 11th week of gestation on the inner hair cells (IHCs) and one week later on the outer hair cells (OHCs). The earliest stages of stereociliary development were similar on both types of cell and were characterized by the presence of round bundles of cilia arising from the surrounding microvilli. A three-dimensional V-shaped arrangement suddenly appeared, accompanied by the disappearance of short cilia on the internal side. Between the 20th and the 22nd weeks of gestation, both types of hair cell had an adult stereociliary pattern, i.e. a rectilinear arrangement on IHCs and W-shaped on OHCs. However, there were signs of immaturity, such as a disarray of OHCs and the presence of the kinocilium, suggesting that the surface of the auditory hair cells achieves its maturation during the last trimester of pregnancy.

pubmed23n0125_13694

Lung of the tree frog, Hyla arborea L. A scanning and transmission electron microscopic study.

Lungs of Hyla arborea L. were examined by scanning and transmission electron microscopy and morphometric methods. The lungs contain several interconnected folds in a netlike reticular arrangement of first, second and third order, mainly covered with pneumocytes. On the septa of first and second order, irregularly distributed small patches of ciliated epithelium devoid of goblet cells are located. Dome-shaped neuroepithelial bodies can be seen in the vicinity of ciliated epithelium. The pulmonary epithelium consists of one type of pneumocyte, which contain in their cytoplasm three kinds of bodies: lamellar, dense and multivesicular. The dense bodies are precursors of lamellar bodies (LBs), while the multivesicular bodies are incorporated into the LBs, being later secreted to the air space. The lining layer covering the internal lung surface contains numerous transformed LBs but tubular myelin figures are scarce. The surface of the lining layer is coated by a thin film. The air-blood barrier, consisting of three layers: epithelium, interstitial space and endothelium, is 0.6 micron thick.

Lung of the tree frog, Hyla arborea L. A scanning and transmission electron microscopic study. Lungs of Hyla arborea L. were examined by scanning and transmission electron microscopy and morphometric methods. The lungs contain several interconnected folds in a netlike reticular arrangement of first, second and third order, mainly covered with pneumocytes. On the septa of first and second order, irregularly distributed small patches of ciliated epithelium devoid of goblet cells are located. Dome-shaped neuroepithelial bodies can be seen in the vicinity of ciliated epithelium. The pulmonary epithelium consists of one type of pneumocyte, which contain in their cytoplasm three kinds of bodies: lamellar, dense and multivesicular. The dense bodies are precursors of lamellar bodies (LBs), while the multivesicular bodies are incorporated into the LBs, being later secreted to the air space. The lining layer covering the internal lung surface contains numerous transformed LBs but tubular myelin figures are scarce. The surface of the lining layer is coated by a thin film. The air-blood barrier, consisting of three layers: epithelium, interstitial space and endothelium, is 0.6 micron thick.

pubmed23n0125_13695

Undulating course of nerve fibres and bands of Fontana in peripheral nerves of the rat.

The undulating course of nerve fibres and the optical effect of that course, i.e. the bands of Fontana, were studied in the peripheral nerves of the adult rat using light microscopy. The arrangement of collagen fibres in the endoneurium of these nerves was evaluated using transmission and scanning electron microscopy. No nerve fibres undulation was noted on the intracranial sections of the cranial nerves or on the spinal roots. In their endoneurium a few, irregularly arranged collagen fibrils were found. In contrast, the nerve fibres undulation and Fontana's bands were a constant feature in the peripheral course of the nerve trunks. They were discernible in vivo and on excised unfixed as well as fixed nerves. The nerve fibres follow a sine-curve course of variable frequency and amplitude. Exposed in vivo, the nerve fibres retained their wave-like course even after removal of the epineurium and perineurium. The endoneurium of these nerves contained numerous undulating longitudinally oriented bundles of collagen fibrils. These findings suggest that the undulating course of the nerve fibres in peripheral nerves is conditional upon the quantity and arrangement of their endoneurial collagen fibrils. When the nerve was stretched in the course of movement, the undulation became straightened out until it disappeared. Conversely, nerve shortening enhanced the undulation. Thus the wave-like alignment of the nerve fibres represents a physiological reserve length for nerve stretching.

Undulating course of nerve fibres and bands of Fontana in peripheral nerves of the rat. The undulating course of nerve fibres and the optical effect of that course, i.e. the bands of Fontana, were studied in the peripheral nerves of the adult rat using light microscopy. The arrangement of collagen fibres in the endoneurium of these nerves was evaluated using transmission and scanning electron microscopy. No nerve fibres undulation was noted on the intracranial sections of the cranial nerves or on the spinal roots. In their endoneurium a few, irregularly arranged collagen fibrils were found. In contrast, the nerve fibres undulation and Fontana's bands were a constant feature in the peripheral course of the nerve trunks. They were discernible in vivo and on excised unfixed as well as fixed nerves. The nerve fibres follow a sine-curve course of variable frequency and amplitude. Exposed in vivo, the nerve fibres retained their wave-like course even after removal of the epineurium and perineurium. The endoneurium of these nerves contained numerous undulating longitudinally oriented bundles of collagen fibrils. These findings suggest that the undulating course of the nerve fibres in peripheral nerves is conditional upon the quantity and arrangement of their endoneurial collagen fibrils. When the nerve was stretched in the course of movement, the undulation became straightened out until it disappeared. Conversely, nerve shortening enhanced the undulation. Thus the wave-like alignment of the nerve fibres represents a physiological reserve length for nerve stretching.

pubmed23n0125_13696

Postnatal development of bovine Sertoli cells.

The fine structure of bovine Sertoli cells was studied from the 4th to the 40th week post natum in order to correlate the progressive acquisition of normal adult morphology with functional development. The considerable increase in tubular size during the first 20 weeks is due to the proliferation of both presumptive Sertoli and germ cells. Aside from this, the presumptive Sertoli cells are seen to expand radially and lengthen considerably. From then on however, the observed increase in tubular diameter during the later period of postnatal development is solely due to the great increase in the number of germ cells. Presumptive Sertoli cells undergo morphological differentiation to mature Sertoli cells during the first 28 weeks of proliferative development. The maturation process includes distinct changes in cell shape, nucleus and cellular organelles, as well as an increase in and differentiation of Sertoli cell surface specializations. At 24 weeks the development of inter-Sertoli cell junctions has reached a point of differentiation where, in our opinion, a functional blood-testis barrier can be expected. During the first 8 weeks an extensive development of rough endoplasmic reticulum and a well-developed Golgi apparatus can be observed, which suggests a high secretory activity in the presumptive Sertoli cells at this time. We speculate that these secretory activities may play a role in the formation of the basal lamina which is extremely well developed during early postnatal life. The subsequent reduction of the basal lamina correlates well with diminished secretory activity in the Sertoli cells.

Postnatal development of bovine Sertoli cells. The fine structure of bovine Sertoli cells was studied from the 4th to the 40th week post natum in order to correlate the progressive acquisition of normal adult morphology with functional development. The considerable increase in tubular size during the first 20 weeks is due to the proliferation of both presumptive Sertoli and germ cells. Aside from this, the presumptive Sertoli cells are seen to expand radially and lengthen considerably. From then on however, the observed increase in tubular diameter during the later period of postnatal development is solely due to the great increase in the number of germ cells. Presumptive Sertoli cells undergo morphological differentiation to mature Sertoli cells during the first 28 weeks of proliferative development. The maturation process includes distinct changes in cell shape, nucleus and cellular organelles, as well as an increase in and differentiation of Sertoli cell surface specializations. At 24 weeks the development of inter-Sertoli cell junctions has reached a point of differentiation where, in our opinion, a functional blood-testis barrier can be expected. During the first 8 weeks an extensive development of rough endoplasmic reticulum and a well-developed Golgi apparatus can be observed, which suggests a high secretory activity in the presumptive Sertoli cells at this time. We speculate that these secretory activities may play a role in the formation of the basal lamina which is extremely well developed during early postnatal life. The subsequent reduction of the basal lamina correlates well with diminished secretory activity in the Sertoli cells.

pubmed23n0125_13697

Influence of colchicine on the distribution of horseradish peroxidase in the secretory ameloblast layer in vitro.

The distribution of exogenous horseradish peroxidase (HRP) in the secretory ameloblast layer of developing rat molar tooth germs was examined in a culture system with and without colchicine. The secretory ameloblast of cultured tooth germs engulfed HRP added to the medium regardless of the presence of colchicine. The reaction product was localized in various vesicles and granules. Without colchicine in the medium, many small vesicles containing HRP were located in the Tomes' processes, whereas only a few were present with colchicine at concentrations above 5 microM. An intense reaction of HRP also appeared in the distal extracellular spaces beyond the distal junctional complexes of ameloblasts cultured without colchicine, whereas it became almost indiscernible in the tooth germs cultured with colchicine. The lack of HRP in the Tomes' processes and distal extracellular spaces of ameloblasts treated with colchicine might be attributed to the disruption of microtubules. The present study suggests that the secretory ameloblast is able to transport tracer molecules through the intracellular pathway from the proximal and lateral extracellular spaces to the distal extracellular space.

Influence of colchicine on the distribution of horseradish peroxidase in the secretory ameloblast layer in vitro. The distribution of exogenous horseradish peroxidase (HRP) in the secretory ameloblast layer of developing rat molar tooth germs was examined in a culture system with and without colchicine. The secretory ameloblast of cultured tooth germs engulfed HRP added to the medium regardless of the presence of colchicine. The reaction product was localized in various vesicles and granules. Without colchicine in the medium, many small vesicles containing HRP were located in the Tomes' processes, whereas only a few were present with colchicine at concentrations above 5 microM. An intense reaction of HRP also appeared in the distal extracellular spaces beyond the distal junctional complexes of ameloblasts cultured without colchicine, whereas it became almost indiscernible in the tooth germs cultured with colchicine. The lack of HRP in the Tomes' processes and distal extracellular spaces of ameloblasts treated with colchicine might be attributed to the disruption of microtubules. The present study suggests that the secretory ameloblast is able to transport tracer molecules through the intracellular pathway from the proximal and lateral extracellular spaces to the distal extracellular space.

pubmed23n0125_13698

Determination of the lengths of nonisotropic linear features in micrographs.

The method described was devised to facilitate rapid and reasonably accurate estimations of the length of a nonisotropic linear feature in a micrograph. It arose from studies in which we were determining the length of the Purkinje cell layer in each of a set of serial sections through the rat cerebellum. The Purkinje cell layer appears as a linear feature in such sections and the simplest and most rapid method of estimating the length of this type of feature is to count the number of intersections that it makes with a series of equally spaced parallel test lines (see, e.g., Weibel, E.R., 1979: Stereological Methods Vol. 1, Practical Methods for Biological Morphometry). In many sections, the Purkinje cell layer was markedly nonisotropic, and the length obtained by this method varied very considerably depending on the orientation of the section relative to the test lines. The present method employs two orthogonal sets of parallel test lines, and the length of the feature is estimated from the square root of the sum of the squares of the counts of the number of intersections with each of the two sets of lines. The result obtained varies very little with the relative orientations of the feature and the test grid and a good estimate of the length can be obtained from the counts from a single random placement of the grid on the section. It has been found that the technique can be carried out efficiently and reliably by relatively inexperienced personnel, and the results are obtained more rapidly than when alternative methods for estimating dimensions of nonisotropic features are used.

Determination of the lengths of nonisotropic linear features in micrographs. The method described was devised to facilitate rapid and reasonably accurate estimations of the length of a nonisotropic linear feature in a micrograph. It arose from studies in which we were determining the length of the Purkinje cell layer in each of a set of serial sections through the rat cerebellum. The Purkinje cell layer appears as a linear feature in such sections and the simplest and most rapid method of estimating the length of this type of feature is to count the number of intersections that it makes with a series of equally spaced parallel test lines (see, e.g., Weibel, E.R., 1979: Stereological Methods Vol. 1, Practical Methods for Biological Morphometry). In many sections, the Purkinje cell layer was markedly nonisotropic, and the length obtained by this method varied very considerably depending on the orientation of the section relative to the test lines. The present method employs two orthogonal sets of parallel test lines, and the length of the feature is estimated from the square root of the sum of the squares of the counts of the number of intersections with each of the two sets of lines. The result obtained varies very little with the relative orientations of the feature and the test grid and a good estimate of the length can be obtained from the counts from a single random placement of the grid on the section. It has been found that the technique can be carried out efficiently and reliably by relatively inexperienced personnel, and the results are obtained more rapidly than when alternative methods for estimating dimensions of nonisotropic features are used.

pubmed23n0125_13699

Osteodentin formation in rat incisor as visualized by radioautography after 3H-proline administration.

Osteodentin formation was studied in rat incisor pulp after adriamycin administration. Male Sprague Dawley rats (100 +/- 5 gm) were injected intravenously with adriamycin (5 mg/kg body weight), and after 7 days they were again injected intravenously with 3H-proline (3 microCi/gm). These animals were killed in groups of three from 5 minutes to 4 hours after proline injection by perfusion with 3% phosphate-buffered formaldehyde followed by 2.5% phosphate-buffered glutaraldehyde. Control animals injected with only physiological saline, and 7 days later with 3H-proline (3 microCi/gm), and were killed at the same time intervals. Radioautography on sections showing osteodentin formation revealed that at 5 minutes after 3H-proline injection the labeling was located over the cells associated with the osteodentin matrix. At 1 hour after injection the labeling was located over the cells and the matrix, while at 4 hours the labeling was seen only over the matrix. It therefore appears that at least a proline-containing component of the osteodentin matrix is synthesized and secreted by the cells associated with it.

Osteodentin formation in rat incisor as visualized by radioautography after 3H-proline administration. Osteodentin formation was studied in rat incisor pulp after adriamycin administration. Male Sprague Dawley rats (100 +/- 5 gm) were injected intravenously with adriamycin (5 mg/kg body weight), and after 7 days they were again injected intravenously with 3H-proline (3 microCi/gm). These animals were killed in groups of three from 5 minutes to 4 hours after proline injection by perfusion with 3% phosphate-buffered formaldehyde followed by 2.5% phosphate-buffered glutaraldehyde. Control animals injected with only physiological saline, and 7 days later with 3H-proline (3 microCi/gm), and were killed at the same time intervals. Radioautography on sections showing osteodentin formation revealed that at 5 minutes after 3H-proline injection the labeling was located over the cells associated with the osteodentin matrix. At 1 hour after injection the labeling was located over the cells and the matrix, while at 4 hours the labeling was seen only over the matrix. It therefore appears that at least a proline-containing component of the osteodentin matrix is synthesized and secreted by the cells associated with it.

pubmed23n0125_13700

Sorting and transepithelial transport of adsorbed protein tracers: effects of temperature.

The epithelium of the guinea pig yolk sac is involved in the selective transport of macromolecules to the fetus. We studied the compartments involved in sorting and transepithelial transport of protein tracers and the effect of lowered temperature (18 degrees C) on these events. Explants of yolk sac were incubated with a mixture of cationized ferritin (CF) and horseradish peroxidase (HRP, Sigma type VI). At 4 degrees C, both tracers were bound to the cell surface and binding of an HRP-gold complex was shown to be inhibited by mannan. At 37 degrees C and 18 degrees C, both tracers were taken up into tubules and vesicles in the apical cytoplasm. Usually the tubules contained a mixture of tracers, but they often showed a polarized distribution with CF and HRP at opposite ends. The vesicles also contained mixtures of the tracers, but some contained only one. In addition, there were some irregularly shaped vacuoles composed of saccules that contained either a mixture, HRP alone, or CF alone. These results suggest that these adsorbed ligands are binding to unique microdomains of the endocytic complex. After 20 min at 37 degrees C coated vesicles 100 nm in diameter were located in the apical cytoplasm and coated vesicles of the same size were located at the lateral cell membrane. Usually they contained only HRP or CF, although occasional mixtures were seen. At 18 degrees C, HRP was transported across the cells in 100 nm vesicles. However, transport of CF was completely inhibited at the lower temperature.(ABSTRACT TRUNCATED AT 250 WORDS)

Sorting and transepithelial transport of adsorbed protein tracers: effects of temperature. The epithelium of the guinea pig yolk sac is involved in the selective transport of macromolecules to the fetus. We studied the compartments involved in sorting and transepithelial transport of protein tracers and the effect of lowered temperature (18 degrees C) on these events. Explants of yolk sac were incubated with a mixture of cationized ferritin (CF) and horseradish peroxidase (HRP, Sigma type VI). At 4 degrees C, both tracers were bound to the cell surface and binding of an HRP-gold complex was shown to be inhibited by mannan. At 37 degrees C and 18 degrees C, both tracers were taken up into tubules and vesicles in the apical cytoplasm. Usually the tubules contained a mixture of tracers, but they often showed a polarized distribution with CF and HRP at opposite ends. The vesicles also contained mixtures of the tracers, but some contained only one. In addition, there were some irregularly shaped vacuoles composed of saccules that contained either a mixture, HRP alone, or CF alone. These results suggest that these adsorbed ligands are binding to unique microdomains of the endocytic complex. After 20 min at 37 degrees C coated vesicles 100 nm in diameter were located in the apical cytoplasm and coated vesicles of the same size were located at the lateral cell membrane. Usually they contained only HRP or CF, although occasional mixtures were seen. At 18 degrees C, HRP was transported across the cells in 100 nm vesicles. However, transport of CF was completely inhibited at the lower temperature.(ABSTRACT TRUNCATED AT 250 WORDS)

pubmed23n0125_13701

The rete ovarii of cattle and deer communicates with the uterine tube.

Textbooks in embryology, histology, and reproductive biology erroneously state that the mammalian rete ovarii consists of a tubular structure in the medulla of the ovary which is closed at both ends. In white-tailed deer and cattle, the lumen of the rete ovarii opens directly into the infundibulum near its attachment to the ovary. This serendipitous discovery followed the serial sectioning of over 100 ovaries. Secretions through this tuboretial opening may affect reproductive processes. Obstructions could explain the development of cysts of clinical importance in both human and veterinary medicine. This rather obscure structure may have more significance than previously recognized.

The rete ovarii of cattle and deer communicates with the uterine tube. Textbooks in embryology, histology, and reproductive biology erroneously state that the mammalian rete ovarii consists of a tubular structure in the medulla of the ovary which is closed at both ends. In white-tailed deer and cattle, the lumen of the rete ovarii opens directly into the infundibulum near its attachment to the ovary. This serendipitous discovery followed the serial sectioning of over 100 ovaries. Secretions through this tuboretial opening may affect reproductive processes. Obstructions could explain the development of cysts of clinical importance in both human and veterinary medicine. This rather obscure structure may have more significance than previously recognized.

pubmed23n0125_13702

Differential distribution of brush cells in the rat lung.

The distribution of brush cells in the rat lung was studied using electron microscopic morphometry. Samples were taken from six distinctive anatomical regions. Tissue from the trachea, lobar bronchi, terminal bronchioles, first alveolar duct bifurcations, proximal alveolar regions, and the distal alveolar region were isolated and embedded in Epox 812. Aside from the trachea and the lobar bronchi, the other four regions were isolated from embedded tissue using microdissection techniques. Electron micrographs taken from thin sections of these samples were analyzed. It was found that brush cells made up 10% of the volume of epithelium covering the first alveolar duct bifurcation. Approximately 2% of the proximal alveolar epithelium, 1.4% of the terminal bronchiolar epithelium, and 3% of the tracheal epithelium were made up of brush cells. No brush-bordered epithelium was found in the lobar bronchi or in the distal alveolar walls. We conclude that brush cells have a distinct spatial location in the lung, being in high concentration in the trachea and in areas where first generation alveolar ducts bifurcate. The highest density was on the bifurcation of the first alveolar ducts, and their density decreased radially from this region.

Differential distribution of brush cells in the rat lung. The distribution of brush cells in the rat lung was studied using electron microscopic morphometry. Samples were taken from six distinctive anatomical regions. Tissue from the trachea, lobar bronchi, terminal bronchioles, first alveolar duct bifurcations, proximal alveolar regions, and the distal alveolar region were isolated and embedded in Epox 812. Aside from the trachea and the lobar bronchi, the other four regions were isolated from embedded tissue using microdissection techniques. Electron micrographs taken from thin sections of these samples were analyzed. It was found that brush cells made up 10% of the volume of epithelium covering the first alveolar duct bifurcation. Approximately 2% of the proximal alveolar epithelium, 1.4% of the terminal bronchiolar epithelium, and 3% of the tracheal epithelium were made up of brush cells. No brush-bordered epithelium was found in the lobar bronchi or in the distal alveolar walls. We conclude that brush cells have a distinct spatial location in the lung, being in high concentration in the trachea and in areas where first generation alveolar ducts bifurcate. The highest density was on the bifurcation of the first alveolar ducts, and their density decreased radially from this region.

pubmed23n0125_13703

Filamentary tubular inclusions in type II pneumocytes of the guinea pig.

The type II pneumocytes of guinea pigs were studied with a transmission electron microscope. It was shown that 5-20% of type II cells contained filamentary tubular inclusions. No reports of similar structures in type II cells of any species have been found in the literature to date. These inclusions measured 90 nm in diameter, varying up to 3 micron in length, and were generally present in the basal and lateral portions of the cells with a close association to the lamellar bodies. They were usually straight but could be curved and tended to aggregate into parallel stacks. In cross-sections, these filamentary tubular inclusions were surrounded by a thickened unit membrane and were composed of microfilaments arranged in circular fashion around a central core. In longitudinal sections, they consisted of rod-like bodies with three lines of filaments. The possible function of the filamentary tubular inclusions is also discussed.

Filamentary tubular inclusions in type II pneumocytes of the guinea pig. The type II pneumocytes of guinea pigs were studied with a transmission electron microscope. It was shown that 5-20% of type II cells contained filamentary tubular inclusions. No reports of similar structures in type II cells of any species have been found in the literature to date. These inclusions measured 90 nm in diameter, varying up to 3 micron in length, and were generally present in the basal and lateral portions of the cells with a close association to the lamellar bodies. They were usually straight but could be curved and tended to aggregate into parallel stacks. In cross-sections, these filamentary tubular inclusions were surrounded by a thickened unit membrane and were composed of microfilaments arranged in circular fashion around a central core. In longitudinal sections, they consisted of rod-like bodies with three lines of filaments. The possible function of the filamentary tubular inclusions is also discussed.

pubmed23n0125_13704

Immunoelectron microscope observations on secretion of human placental lactogen (hPL) in the human chorionic villi.

Immunoelectron microscope localization of human placental lactogen (hPL) was investigated in the chorionic villi from week 7 to full-term gestation with the protein A-gold technique. With specific antiserum against hPL, immuno-reactive gold particles were found to be preferentially located in Golgi-derived, electron-dense small granules of 80-180 nm in the syncytiotrophoblast. Our electron micrographs indicate that these small granules increase in number in the course of gestation and are released by exocytosis from the apical cell surface. The present study reveals that hPL is segregated from the Golgi apparatus, stored in the syncytiotrophoblast as secretory granules, and released into the maternal blood.

Immunoelectron microscope observations on secretion of human placental lactogen (hPL) in the human chorionic villi. Immunoelectron microscope localization of human placental lactogen (hPL) was investigated in the chorionic villi from week 7 to full-term gestation with the protein A-gold technique. With specific antiserum against hPL, immuno-reactive gold particles were found to be preferentially located in Golgi-derived, electron-dense small granules of 80-180 nm in the syncytiotrophoblast. Our electron micrographs indicate that these small granules increase in number in the course of gestation and are released by exocytosis from the apical cell surface. The present study reveals that hPL is segregated from the Golgi apparatus, stored in the syncytiotrophoblast as secretory granules, and released into the maternal blood.

pubmed23n0125_13705

Venous distribution of superficial cervical region in rhesus (Macaca mulatta) monkey.

The superficial veins of the cervical region in over 50 Macaca mulatta monkeys were studied. We found, in addition to the external jugular vein, another major vein, which we have termed jugular accessory. It is comparable in size and runs ventral to the external jugular vein. It commenced at the angle of the mouth, ran in a groove on the dorsal aspect of the submandibular gland, and descended on the surface of the sternocleidomastoid muscle where it was connected to the external jugular vein by a short transverse twig. It then descended toward the clavicle, crossed it ventrally, and immediately joined the cephalic vein. The resultant common vein pierced the thoracic wall between the clavicle and first rib and joined the external jugular and axillary veins, producing the subclavian vein. It was the jugular accessory and the external jugular, being connected as described, that formed an "H"-shaped system.

Venous distribution of superficial cervical region in rhesus (Macaca mulatta) monkey. The superficial veins of the cervical region in over 50 Macaca mulatta monkeys were studied. We found, in addition to the external jugular vein, another major vein, which we have termed jugular accessory. It is comparable in size and runs ventral to the external jugular vein. It commenced at the angle of the mouth, ran in a groove on the dorsal aspect of the submandibular gland, and descended on the surface of the sternocleidomastoid muscle where it was connected to the external jugular vein by a short transverse twig. It then descended toward the clavicle, crossed it ventrally, and immediately joined the cephalic vein. The resultant common vein pierced the thoracic wall between the clavicle and first rib and joined the external jugular and axillary veins, producing the subclavian vein. It was the jugular accessory and the external jugular, being connected as described, that formed an "H"-shaped system.

pubmed23n0125_13706

Morphological evaluation of vascular smooth muscle cell: length and width from a single scanning electron micrograph of microvessels.

Accurate three-dimensional data on the structure of vascular smooth muscle cells is essential for understanding the microvascular system in both normal or disease conditions. The laborious serial reconstruction methods have limited the amount of data collected on the structure of individual vascular smooth muscle (VSM) cells. The circumferential viewing of whole vessel segments via scanning electron microscopy provides an alternative approach, but even this technique is highly specialized and tedious. This study presents a simplified method to determine the average cell length and width of individual VSM cells by using only one view of a microvessel (single view). The vessels do not have to be isolated for circumferential viewing and can be left in their host tissue if desired. Values for the average VSM cell length and width were obtained by both circumferential- and single-view approach on the same vessels. The average cell length and width obtained from the single-view method (using one-third circumference) duplicated the mean length and width measurements obtained by circumferential viewing.

Morphological evaluation of vascular smooth muscle cell: length and width from a single scanning electron micrograph of microvessels. Accurate three-dimensional data on the structure of vascular smooth muscle cells is essential for understanding the microvascular system in both normal or disease conditions. The laborious serial reconstruction methods have limited the amount of data collected on the structure of individual vascular smooth muscle (VSM) cells. The circumferential viewing of whole vessel segments via scanning electron microscopy provides an alternative approach, but even this technique is highly specialized and tedious. This study presents a simplified method to determine the average cell length and width of individual VSM cells by using only one view of a microvessel (single view). The vessels do not have to be isolated for circumferential viewing and can be left in their host tissue if desired. Values for the average VSM cell length and width were obtained by both circumferential- and single-view approach on the same vessels. The average cell length and width obtained from the single-view method (using one-third circumference) duplicated the mean length and width measurements obtained by circumferential viewing.

pubmed23n0125_13707

Comparative toxicity of atracurium and metocurine in isolated rat hepatocytes.

Primary cultures of liver cells isolated from seven rats were used to study the possible toxicity of atracurium and metocurine. The muscle relaxants were separately added to the culture medium and the cells then incubated for 4 hr. The amount of lactic dehydrogenase (LDH) that leaked into the culture medium was determined at the end of incubation. The customary assumption was made that the exudation of LDH reflects the toxic effects of the relaxants. In untreated dishes, approximately 11% of the total intracellular LDH leaked out during the incubation. The net leakage of LDH produced by the relaxants was obtained by subtracting this amount from the LDH activity determined in the media of dishes with the relaxants added. On this basis, metocurine, in concentrations of 12-850 X 10(-6)M, did not cause a net leak of LDH. On the other hand, atracurium, in similar molar concentrations, caused a statistically significant and concentration-dependent leak of LDH that, at its maximum, amounted to more than one half of the intracellular LDH. The results are interpreted in terms of damage to cellular membranes produced by atracurium or its metabolites. Although the exact biochemical process was not identified, we hypothesize that acrylates--produced by Hofmann elimination from atracurium--might be the likely toxic species.

Comparative toxicity of atracurium and metocurine in isolated rat hepatocytes. Primary cultures of liver cells isolated from seven rats were used to study the possible toxicity of atracurium and metocurine. The muscle relaxants were separately added to the culture medium and the cells then incubated for 4 hr. The amount of lactic dehydrogenase (LDH) that leaked into the culture medium was determined at the end of incubation. The customary assumption was made that the exudation of LDH reflects the toxic effects of the relaxants. In untreated dishes, approximately 11% of the total intracellular LDH leaked out during the incubation. The net leakage of LDH produced by the relaxants was obtained by subtracting this amount from the LDH activity determined in the media of dishes with the relaxants added. On this basis, metocurine, in concentrations of 12-850 X 10(-6)M, did not cause a net leak of LDH. On the other hand, atracurium, in similar molar concentrations, caused a statistically significant and concentration-dependent leak of LDH that, at its maximum, amounted to more than one half of the intracellular LDH. The results are interpreted in terms of damage to cellular membranes produced by atracurium or its metabolites. Although the exact biochemical process was not identified, we hypothesize that acrylates--produced by Hofmann elimination from atracurium--might be the likely toxic species.

pubmed23n0125_13708

Preoperative oral fluids: is a five-hour fast justified prior to elective surgery?

The effects of preoperative oral administration of 150 ml fluid were studied prospectively in 140 unpremedicated, ambulatory outpatients presenting for first trimester therapeutic abortion. Intraoperative gastric fluid volume, pH, and rate of gastric emptying were measured in the four groups to which patients were randomly assigned. At an average time of 2 1/2 hr preoperatively all patients received either oral ranitidine, 150 mg, or a placebo tablet, with the nonabsorbable marker dye bromosulphthalein (BSP, 50 mg in 10 ml water, followed by either 150 ml water or no further fluid. The effect of volume ingested was assessed by comparing the volume of gastric contents obtained by gastric tube suctioning at the completion of surgery in the two groups given placebos. The gastric volume was significantly less in patients given 150 ml water (17.6 +/- 14.5) than in those given only BSP (26.7 +/- 18.9) (P less than 0.02), and was further significantly decreased in the two groups given ranitidine (8.3 +/- 7.3, 9.5 +/- 7.7 ml) (P less than 0.001). Mean pH values were significantly higher in the two ranitidine groups (5.52 +/- 1.79, 5.03 +/- 1.79) than in the two placebo groups (1.75 +/- 0.94, 1.92 +/- 1.27). The combination of a residual volume of 25 ml and pH less than 2.5 was found in 46% of patients given only BSP with placebo, in 23% of those given 150 ml water with placebo, and in no patient given ranitidine. There was no correlation between the gastric volume or pH values with the ingestion-surgery interval in patients given 150 ml water.

Preoperative oral fluids: is a five-hour fast justified prior to elective surgery? The effects of preoperative oral administration of 150 ml fluid were studied prospectively in 140 unpremedicated, ambulatory outpatients presenting for first trimester therapeutic abortion. Intraoperative gastric fluid volume, pH, and rate of gastric emptying were measured in the four groups to which patients were randomly assigned. At an average time of 2 1/2 hr preoperatively all patients received either oral ranitidine, 150 mg, or a placebo tablet, with the nonabsorbable marker dye bromosulphthalein (BSP, 50 mg in 10 ml water, followed by either 150 ml water or no further fluid. The effect of volume ingested was assessed by comparing the volume of gastric contents obtained by gastric tube suctioning at the completion of surgery in the two groups given placebos. The gastric volume was significantly less in patients given 150 ml water (17.6 +/- 14.5) than in those given only BSP (26.7 +/- 18.9) (P less than 0.02), and was further significantly decreased in the two groups given ranitidine (8.3 +/- 7.3, 9.5 +/- 7.7 ml) (P less than 0.001). Mean pH values were significantly higher in the two ranitidine groups (5.52 +/- 1.79, 5.03 +/- 1.79) than in the two placebo groups (1.75 +/- 0.94, 1.92 +/- 1.27). The combination of a residual volume of 25 ml and pH less than 2.5 was found in 46% of patients given only BSP with placebo, in 23% of those given 150 ml water with placebo, and in no patient given ranitidine. There was no correlation between the gastric volume or pH values with the ingestion-surgery interval in patients given 150 ml water.

pubmed23n0125_13709

Isoflurane decreases the cortisol response to cardiopulmonary bypass.

Eighteen patients with normal left ventricular function scheduled for elective myocardial revascularization were anesthetized with fentanyl (52-58 micrograms/kg). At the beginning of hypothermic cardiopulmonary bypass (CPB) they were assigned to a control (C) group (n = 6) that did not receive further anesthesia, or to a group given either 1% isoflurane (n = 6) or 2% isoflurane (n = 6). Blood samples for measurement of total plasma cortisol concentrations were obtained before, during, and after CPB. Hemodynamic measurements before and after CPB were not different among groups. Patients in group C required higher infusion rates of sodium nitroprusside (P less than or equal to 0.05) and patients given 2% isoflurane received more phenylephrine (P less than or equal to 0.05) to keep mean arterial pressure at 50 +/- 10 mm Hg during CPB. Isoflurane caused a dose-related decrease in total plasma cortisol concentrations during and after CPB. We conclude that increased depth of anesthesia attenuates the cortisol (stress) response to cardiopulmonary bypass.

Isoflurane decreases the cortisol response to cardiopulmonary bypass. Eighteen patients with normal left ventricular function scheduled for elective myocardial revascularization were anesthetized with fentanyl (52-58 micrograms/kg). At the beginning of hypothermic cardiopulmonary bypass (CPB) they were assigned to a control (C) group (n = 6) that did not receive further anesthesia, or to a group given either 1% isoflurane (n = 6) or 2% isoflurane (n = 6). Blood samples for measurement of total plasma cortisol concentrations were obtained before, during, and after CPB. Hemodynamic measurements before and after CPB were not different among groups. Patients in group C required higher infusion rates of sodium nitroprusside (P less than or equal to 0.05) and patients given 2% isoflurane received more phenylephrine (P less than or equal to 0.05) to keep mean arterial pressure at 50 +/- 10 mm Hg during CPB. Isoflurane caused a dose-related decrease in total plasma cortisol concentrations during and after CPB. We conclude that increased depth of anesthesia attenuates the cortisol (stress) response to cardiopulmonary bypass.

pubmed23n0125_13710

Effects of ketamine on low intensity tactile sensory input are not dependent upon a spinal site of action.

The development of a technique for studying spinal dorsal horn electrophysiology in intact, awake, drug-free cats enables the study of spinal sites and mechanisms of action of anesthetic and analgesic agents in a system that more closely reflects normal physiology. Using this technique, we reevaluated the effect of ketamine on spinal dorsal horn sensory transmission. The results of our study confirm previous work done in acute preparations. Ketamine (maximum dose 20 mg/kg) did not significantly reduce the response of low threshold (n = 11) dorsal horn neurons to low intensity sensory stimulation. However, that same dose of ketamine did suppress noxiously evoked activity of the two wide dynamic range neurons encountered in the study, while having a varied effect on neurons responsive to proprioceptive input (n = 7). These findings confirm that, in the intact animal with all modulatory systems intact, ketamine "dissociation" of low intensity tactile stimuli does not appear to involve a spinal mechanism of action. The results also support the importance of spinal sites of action for the analgesia produced by ketamine, as well as the importance of distinguishing between the anesthetic and analgesic effects of that drug.

Effects of ketamine on low intensity tactile sensory input are not dependent upon a spinal site of action. The development of a technique for studying spinal dorsal horn electrophysiology in intact, awake, drug-free cats enables the study of spinal sites and mechanisms of action of anesthetic and analgesic agents in a system that more closely reflects normal physiology. Using this technique, we reevaluated the effect of ketamine on spinal dorsal horn sensory transmission. The results of our study confirm previous work done in acute preparations. Ketamine (maximum dose 20 mg/kg) did not significantly reduce the response of low threshold (n = 11) dorsal horn neurons to low intensity sensory stimulation. However, that same dose of ketamine did suppress noxiously evoked activity of the two wide dynamic range neurons encountered in the study, while having a varied effect on neurons responsive to proprioceptive input (n = 7). These findings confirm that, in the intact animal with all modulatory systems intact, ketamine "dissociation" of low intensity tactile stimuli does not appear to involve a spinal mechanism of action. The results also support the importance of spinal sites of action for the analgesia produced by ketamine, as well as the importance of distinguishing between the anesthetic and analgesic effects of that drug.

pubmed23n0125_13711

Effects of halothane and decreased PO2 on high energy phosphate levels maintained by isolated rat liver mitochondria.

Steady states of oxidative phosphorylation were achieved in mitochondrial suspensions continuously equilibrated with constant gas mixtures, simulating the conditions under which mitochondria contribute to the cellular energy status in vivo. The dependence of the mitochondria-maintained adenosine triphosphate/adenosine diphosphate (ATP/ADP) ratio on oxygen and halothane levels was examined at predetermined, clinically relevant concentrations of both gases. Inclusion of 1% halothane in the gas mixture decreased ATP/ADP by about half when mitochondrial respiration was supported by NAD-linked substrate (glutamate); succinate-supported ATP/ADP was not inhibited. With either substrate, and whether or not 1% halothane was present. ATP/ADP was unaffected by decreases in PO2 to values as low as 1.6 mm Hg. Under a range of typical in vivo conditions, therefore, 1% halothane significantly inhibited the mitochondrial contribution to steady state energy balance, whereas decreases in PO2 did not. Combined effects of 1% halothane and reduced PO2 on ATP/ADP were not seen, i.e., halothane did not increase the critical PO2 level (hypoxic threshold) for inhibition of mitochondrial ATP production.

Effects of halothane and decreased PO2 on high energy phosphate levels maintained by isolated rat liver mitochondria. Steady states of oxidative phosphorylation were achieved in mitochondrial suspensions continuously equilibrated with constant gas mixtures, simulating the conditions under which mitochondria contribute to the cellular energy status in vivo. The dependence of the mitochondria-maintained adenosine triphosphate/adenosine diphosphate (ATP/ADP) ratio on oxygen and halothane levels was examined at predetermined, clinically relevant concentrations of both gases. Inclusion of 1% halothane in the gas mixture decreased ATP/ADP by about half when mitochondrial respiration was supported by NAD-linked substrate (glutamate); succinate-supported ATP/ADP was not inhibited. With either substrate, and whether or not 1% halothane was present. ATP/ADP was unaffected by decreases in PO2 to values as low as 1.6 mm Hg. Under a range of typical in vivo conditions, therefore, 1% halothane significantly inhibited the mitochondrial contribution to steady state energy balance, whereas decreases in PO2 did not. Combined effects of 1% halothane and reduced PO2 on ATP/ADP were not seen, i.e., halothane did not increase the critical PO2 level (hypoxic threshold) for inhibition of mitochondrial ATP production.

pubmed23n0125_13712

Genetic predisposition to liver damage after halothane anesthesia in guinea pigs.

Three 4-hr normoxic (21% oxygen) exposures to 1% halothane administered 3 days apart were associated with elevations in serum alanine aminotransferase (ALT) activity in four of 20 guinea pigs after the initial and third exposures. Serum alanine aminotransferase values were not measured after the second anesthetic. Susceptibility was defined as an ALT level greater than 300 IU/L after halothane. Nonsusceptible animals, that is, animals without significant increases in ALT values after halothane, remained nonsusceptible after reexposure. Serum alanine aminotransferase values after the first and third anesthesias were significantly correlated (rs = 0.86, P less than 0.001). Two exposures of another 30 guinea pigs at a 5-week interval resulted in high elevations of ALT in the same eight animals after both anesthetics. In contrast, after an initial exposure nonsusceptible animals remained nonsusceptible upon reexposure. Serum alanine aminotransferase levels after the first and second anesthetics were significantly correlated (rs = 0.85, P less than 0.001). The proportion of first generation (F1) males with elevated ALTs whose parents were susceptible to halothane hepatotoxicity (HH) was significantly higher than the proportion of males with elevated ALTs in a random group of 90 males (P less than 0.005). First generation males and females of nonsusceptible parents had ALTs within the normal range after halothane exposure. These studies suggest that in the guinea pig genetic predisposition is an important determinant of susceptibility to HH, although other contributing factors are not excluded.

Genetic predisposition to liver damage after halothane anesthesia in guinea pigs. Three 4-hr normoxic (21% oxygen) exposures to 1% halothane administered 3 days apart were associated with elevations in serum alanine aminotransferase (ALT) activity in four of 20 guinea pigs after the initial and third exposures. Serum alanine aminotransferase values were not measured after the second anesthetic. Susceptibility was defined as an ALT level greater than 300 IU/L after halothane. Nonsusceptible animals, that is, animals without significant increases in ALT values after halothane, remained nonsusceptible after reexposure. Serum alanine aminotransferase values after the first and third anesthesias were significantly correlated (rs = 0.86, P less than 0.001). Two exposures of another 30 guinea pigs at a 5-week interval resulted in high elevations of ALT in the same eight animals after both anesthetics. In contrast, after an initial exposure nonsusceptible animals remained nonsusceptible upon reexposure. Serum alanine aminotransferase levels after the first and second anesthetics were significantly correlated (rs = 0.85, P less than 0.001). The proportion of first generation (F1) males with elevated ALTs whose parents were susceptible to halothane hepatotoxicity (HH) was significantly higher than the proportion of males with elevated ALTs in a random group of 90 males (P less than 0.005). First generation males and females of nonsusceptible parents had ALTs within the normal range after halothane exposure. These studies suggest that in the guinea pig genetic predisposition is an important determinant of susceptibility to HH, although other contributing factors are not excluded.

pubmed23n0125_13713

Morphine and fentanyl interactions with thiopental in relation to movement response to noxious stimulation.

The effects of morphine-thiopental and fentanyl-thiopental combinations on the movement response caused by tail clamping were studied in rats. Doses that prevented movement response when the agents were given singly and when the agents were given in combination were determined by a probit procedure and compared with isobolographic analysis. With doses of the above agents sufficient to block the movement response to tail clamping (ED50 values: 4.7 (3.3-6.6) mg/kg intravenously for morphine; 8.3 (5.8-11.3) micrograms/kg intravenously for fentanyl; and 18.8 (17.9-19.7) mg/kg intravenously for thiopental) both fentanyl and, to a lesser extent, morphine have a less than additive or an antagonistic interaction with thiopental. This antagonism is a relative one that does not increase the requirement for one agent upon the addition of another agent.

Morphine and fentanyl interactions with thiopental in relation to movement response to noxious stimulation. The effects of morphine-thiopental and fentanyl-thiopental combinations on the movement response caused by tail clamping were studied in rats. Doses that prevented movement response when the agents were given singly and when the agents were given in combination were determined by a probit procedure and compared with isobolographic analysis. With doses of the above agents sufficient to block the movement response to tail clamping (ED50 values: 4.7 (3.3-6.6) mg/kg intravenously for morphine; 8.3 (5.8-11.3) micrograms/kg intravenously for fentanyl; and 18.8 (17.9-19.7) mg/kg intravenously for thiopental) both fentanyl and, to a lesser extent, morphine have a less than additive or an antagonistic interaction with thiopental. This antagonism is a relative one that does not increase the requirement for one agent upon the addition of another agent.

pubmed23n0125_13714

The in vitro and in vivo effects of isoflurane and nitrous oxide on platelet aggregation.

In view of the possible antiaggregation effects of newer general anesthetics we investigated the in vitro and in vivo effects of isoflurane and nitrous oxide on platelet aggregation. Platelets obtained from 18 healthy volunteers, were exposed in vitro for 30 min in a closed chamber to oxygen-carbon dioxide (90%, 5%) (control), oxygen-carbon dioxide-nitrous oxide (80%), or oxygen-carbon dioxide-isoflurane (1.5%) with or without nitrous oxide (80%). Samples were tested in ADP- and collagen-induced aggregation tests. Both nitrous oxide and isoflurane produced statistically significant inhibition of ADP-induced aggregation. Inhibition of collagen-induced aggregation was not statistically significant. In 18 patients who received nitrous oxide (3 L/min) and isoflurane (1-2%) during anesthesia, platelet aggregation was reduced significantly. We conclude that both nitrous oxide and isoflurane cause moderate but statistically significant inhibition of platelet aggregation that may be clinically important in some patients.

The in vitro and in vivo effects of isoflurane and nitrous oxide on platelet aggregation. In view of the possible antiaggregation effects of newer general anesthetics we investigated the in vitro and in vivo effects of isoflurane and nitrous oxide on platelet aggregation. Platelets obtained from 18 healthy volunteers, were exposed in vitro for 30 min in a closed chamber to oxygen-carbon dioxide (90%, 5%) (control), oxygen-carbon dioxide-nitrous oxide (80%), or oxygen-carbon dioxide-isoflurane (1.5%) with or without nitrous oxide (80%). Samples were tested in ADP- and collagen-induced aggregation tests. Both nitrous oxide and isoflurane produced statistically significant inhibition of ADP-induced aggregation. Inhibition of collagen-induced aggregation was not statistically significant. In 18 patients who received nitrous oxide (3 L/min) and isoflurane (1-2%) during anesthesia, platelet aggregation was reduced significantly. We conclude that both nitrous oxide and isoflurane cause moderate but statistically significant inhibition of platelet aggregation that may be clinically important in some patients.

pubmed23n0125_13715

Tourniquet pain: a volunteer study.

The effect of inflation pressure (300 and 400 mm Hg) and method of exsanguination (gravity and Esmarch bandage) on the time of onset and the severity of tourniquet-induced pain in the lower extremity was investigated in 11 unmedicated adult volunteers. Each volunteer underwent eight experiments in a random order. A visual analog scale was used to assess pain and discomfort. Blood pressure and pulse rate were measured continuously. Experiments were concluded when the pain rose to a prefixed level. All experiments were performed using a standard orthopedic tourniquet (7 cm wide). Ten additional experiments were carried out using a Bier blockade tourniquet (5 cm wide). There were no differences in duration of tourniquet inflation between inflation pressures nor between methods of exsanguination. There was a small and transient but nevertheless statistically significant increase in blood pressure caused by inflation and a significantly larger increase just before deflation. The 5-cm tourniquet experiments, otherwise identical to the 7-cm tourniquet experiments, were tolerated significantly longer due to a longer time of onset and less severe pain. The 5-cm tourniquet also needed significantly higher inflation pressures to fully occlude the arterial supply (240-450 mm Hg). In all instances, 260 mm Hg was adequate to fully occlude the arterial supply when a 7-cm tourniquet was used. Only half of the experiments were concluded due to intolerable pain at the site of the tourniquet. Most of the others were concluded due to pain mainly in the calf or pain throughout the leg.(ABSTRACT TRUNCATED AT 250 WORDS)

Tourniquet pain: a volunteer study. The effect of inflation pressure (300 and 400 mm Hg) and method of exsanguination (gravity and Esmarch bandage) on the time of onset and the severity of tourniquet-induced pain in the lower extremity was investigated in 11 unmedicated adult volunteers. Each volunteer underwent eight experiments in a random order. A visual analog scale was used to assess pain and discomfort. Blood pressure and pulse rate were measured continuously. Experiments were concluded when the pain rose to a prefixed level. All experiments were performed using a standard orthopedic tourniquet (7 cm wide). Ten additional experiments were carried out using a Bier blockade tourniquet (5 cm wide). There were no differences in duration of tourniquet inflation between inflation pressures nor between methods of exsanguination. There was a small and transient but nevertheless statistically significant increase in blood pressure caused by inflation and a significantly larger increase just before deflation. The 5-cm tourniquet experiments, otherwise identical to the 7-cm tourniquet experiments, were tolerated significantly longer due to a longer time of onset and less severe pain. The 5-cm tourniquet also needed significantly higher inflation pressures to fully occlude the arterial supply (240-450 mm Hg). In all instances, 260 mm Hg was adequate to fully occlude the arterial supply when a 7-cm tourniquet was used. Only half of the experiments were concluded due to intolerable pain at the site of the tourniquet. Most of the others were concluded due to pain mainly in the calf or pain throughout the leg.(ABSTRACT TRUNCATED AT 250 WORDS)

pubmed23n0125_13716

Addition of glucose to bupivacaine in spinal anesthesia increases incidence of tourniquet pain.

The effect of baricity of 0.5% bupivacaine on the incidence of tourniquet pain when used for spinal anesthesia was evaluated in 60 patients undergoing orthopedic surgery. Three ml of either hyperbaric (8% glucose) or isobaric (glucose-free) solution was used. A standard 7-cm orthopedic tourniquet was applied at the thigh and was inflated to 300 mm Hg for 2 hr or until the patient experienced pain from the tourniquet. During application time, the levels of sensory block to pin prick were similar in the groups. The incidence of tourniquet pain in the glucose-free group (4/30) was significantly lower than in the hyperbaric group (11/30).

Addition of glucose to bupivacaine in spinal anesthesia increases incidence of tourniquet pain. The effect of baricity of 0.5% bupivacaine on the incidence of tourniquet pain when used for spinal anesthesia was evaluated in 60 patients undergoing orthopedic surgery. Three ml of either hyperbaric (8% glucose) or isobaric (glucose-free) solution was used. A standard 7-cm orthopedic tourniquet was applied at the thigh and was inflated to 300 mm Hg for 2 hr or until the patient experienced pain from the tourniquet. During application time, the levels of sensory block to pin prick were similar in the groups. The incidence of tourniquet pain in the glucose-free group (4/30) was significantly lower than in the hyperbaric group (11/30).

pubmed23n0125_13717

Effect of short-term smoking halt on carboxyhemoglobin levels and P50 values.

Fifteen informed volunteers who smoked one to two packs of cigarettes per day were studied to measure changes in carboxyhemoglobin levels and P50 after smoking was stopped for 12 hr. Before smoking was stopped, carboxyhemoglobin levels (6.55 +/- 0.40%) were above normal, and the P50 (22.92 +/- 0.25 mm Hg) was significantly shifted to the left. After smoking was stopped for 12 hr there was a significant decrease in carboxyhemoglobin levels to 1.06 +/- 0.16% (P less than 0.001), and P50 shifted towards normal to 26.41 +/- 0.14 mm Hg (P less than 0.001). The authors conclude that within even as little as 12 hr after cessation of smoking, carboxyhemoglobin and P50 levels change towards normal values.

Effect of short-term smoking halt on carboxyhemoglobin levels and P50 values. Fifteen informed volunteers who smoked one to two packs of cigarettes per day were studied to measure changes in carboxyhemoglobin levels and P50 after smoking was stopped for 12 hr. Before smoking was stopped, carboxyhemoglobin levels (6.55 +/- 0.40%) were above normal, and the P50 (22.92 +/- 0.25 mm Hg) was significantly shifted to the left. After smoking was stopped for 12 hr there was a significant decrease in carboxyhemoglobin levels to 1.06 +/- 0.16% (P less than 0.001), and P50 shifted towards normal to 26.41 +/- 0.14 mm Hg (P less than 0.001). The authors conclude that within even as little as 12 hr after cessation of smoking, carboxyhemoglobin and P50 levels change towards normal values.

pubmed23n0125_13718

Cerebral arteriovenous oxygen content difference during barbiturate therapy in patients with acute brain damage.

This study evaluated the reliability of cerebral blood flow equivalent (CBFE), which was calculated as the reciprocal of cerebral arteriovenous oxygen content difference (C(av)DO2) as a monitor during barbiturate therapy in patients with cerebral ischemic insults. A barbiturate (thiamylal) was administered at a rate of 3 mg . kg-1 . hr-1 for 2-5 days to four patients who had suffered cardiac arrest, four with acute focal ischemia, two with postoperative brain edema after neurosurgery, and one with brain damage due to asphyxia. Four of the 11 patients completely recovered neurologically (recovery group), and others had neurological sequelae or died (nonrecovery group). The mean value of CBFE in the recovery group decreased significantly with barbiturate therapy to 13 +/- 1 ml blood/ml O2 from 39 +/- 3 ml blood/ml O2 but did not decrease in the nonrecovery group. We conclude that CBFE can be useful for monitoring the effect of barbiturate therapy in ischemic brain insults.

Cerebral arteriovenous oxygen content difference during barbiturate therapy in patients with acute brain damage. This study evaluated the reliability of cerebral blood flow equivalent (CBFE), which was calculated as the reciprocal of cerebral arteriovenous oxygen content difference (C(av)DO2) as a monitor during barbiturate therapy in patients with cerebral ischemic insults. A barbiturate (thiamylal) was administered at a rate of 3 mg . kg-1 . hr-1 for 2-5 days to four patients who had suffered cardiac arrest, four with acute focal ischemia, two with postoperative brain edema after neurosurgery, and one with brain damage due to asphyxia. Four of the 11 patients completely recovered neurologically (recovery group), and others had neurological sequelae or died (nonrecovery group). The mean value of CBFE in the recovery group decreased significantly with barbiturate therapy to 13 +/- 1 ml blood/ml O2 from 39 +/- 3 ml blood/ml O2 but did not decrease in the nonrecovery group. We conclude that CBFE can be useful for monitoring the effect of barbiturate therapy in ischemic brain insults.

pubmed23n0125_13719

Pulmonary and systemic hemodynamic effects of nitrous oxide in infants with normal and elevated pulmonary vascular resistance.

The hemodynamic response to 50% nitrous oxide was studied in 12 sedated but responsive infants in the intensive care unit following repair of their congenital heart disease. One-half of the infants studied had an elevated pulmonary vascular resistance index (PVRI greater than 3.5 Wood units). During mechanical ventilation with a fractional inspired O2 concentration (FIO2) of 0.5, hemodynamic parameters were measured after equilibration with 50% nitrogen and then after 50% nitrous oxide. The sequence was repeated once to assure reproducibility of the responses. Average heart rate decreased by 9%, mean arterial blood pressure decreased by 12%, and cardiac index decreased by 13% in both the elevated and normal PVRI groups each time nitrous oxide was given. Although statistically significant, these changes would not generally be clinically important except in infants with severely compromised cardiovascular reserve. In contrast, pulmonary artery pressure and PVRI were not significantly changed by administration of 50% nitrous oxide in either the group with normal PVRI or the group with preexisting elevated PVRI. We conclude that while these mild depressant effects of nitrous oxide on systemic hemodynamics in infants are similar to those previously reported in adults, in infants nitrous oxide does not produce the elevations in pulmonary artery pressure and pulmonary vascular resistance seen in adults.

Pulmonary and systemic hemodynamic effects of nitrous oxide in infants with normal and elevated pulmonary vascular resistance. The hemodynamic response to 50% nitrous oxide was studied in 12 sedated but responsive infants in the intensive care unit following repair of their congenital heart disease. One-half of the infants studied had an elevated pulmonary vascular resistance index (PVRI greater than 3.5 Wood units). During mechanical ventilation with a fractional inspired O2 concentration (FIO2) of 0.5, hemodynamic parameters were measured after equilibration with 50% nitrogen and then after 50% nitrous oxide. The sequence was repeated once to assure reproducibility of the responses. Average heart rate decreased by 9%, mean arterial blood pressure decreased by 12%, and cardiac index decreased by 13% in both the elevated and normal PVRI groups each time nitrous oxide was given. Although statistically significant, these changes would not generally be clinically important except in infants with severely compromised cardiovascular reserve. In contrast, pulmonary artery pressure and PVRI were not significantly changed by administration of 50% nitrous oxide in either the group with normal PVRI or the group with preexisting elevated PVRI. We conclude that while these mild depressant effects of nitrous oxide on systemic hemodynamics in infants are similar to those previously reported in adults, in infants nitrous oxide does not produce the elevations in pulmonary artery pressure and pulmonary vascular resistance seen in adults.

pubmed23n0125_13720

Energy deficits in hepatocytes isolated from phenobarbital-treated or fasted rats and briefly exposed to halothane and hypoxia in vitro.

Experimental factors implicated in the pathogenesis of halothane hepatotoxicity in the phenobarbital-hypoxia rat model were examined for direct effects on the energy status of isolated rat liver cells in vitro. Intact hepatocytes were isolated after collagenase perfusion of livers of adult male Fischer 344 rats previously treated with phenobarbital (0.1% in drinking water for 5-7 days) and/or deprived of food for 48 h. Cells were incubated in Krebs-Henseleit buffer + substrates for 10 min at steady states of energy metabolism, with extracellular PO2 constant at 32, 16, or 4 mmHg +/- 1% halothane. Fasting produced the largest energy deficits in incubated hepatocytes, regardless of phenobarbital treatment status, PO2 value, or presence/absence of halothane. The combination of hypoxic PO2 (4 mmHg) and 1% halothane shifted lactate metabolism toward lactate production, whereas hypoxia or halothane alone did not. Prior phenobarbital treatment plus hypoxia decreased adenosine triphosphate/adenosine diphosphate (ATP/ADP) and increased lactate production compared with drug treatment or hypoxia alone. We conclude that pathogenic factors that interact to produce halothane hepatotoxicity act directly and jointly on isolated liver cells to produce energy deficits within 10 min. Differences in the relative importance of pathogenic factors in vitro and in vivo suggest that short-term, direct effects on hepatocellular energy status are not solely responsible for halothane hepatotoxicity.

Energy deficits in hepatocytes isolated from phenobarbital-treated or fasted rats and briefly exposed to halothane and hypoxia in vitro. Experimental factors implicated in the pathogenesis of halothane hepatotoxicity in the phenobarbital-hypoxia rat model were examined for direct effects on the energy status of isolated rat liver cells in vitro. Intact hepatocytes were isolated after collagenase perfusion of livers of adult male Fischer 344 rats previously treated with phenobarbital (0.1% in drinking water for 5-7 days) and/or deprived of food for 48 h. Cells were incubated in Krebs-Henseleit buffer + substrates for 10 min at steady states of energy metabolism, with extracellular PO2 constant at 32, 16, or 4 mmHg +/- 1% halothane. Fasting produced the largest energy deficits in incubated hepatocytes, regardless of phenobarbital treatment status, PO2 value, or presence/absence of halothane. The combination of hypoxic PO2 (4 mmHg) and 1% halothane shifted lactate metabolism toward lactate production, whereas hypoxia or halothane alone did not. Prior phenobarbital treatment plus hypoxia decreased adenosine triphosphate/adenosine diphosphate (ATP/ADP) and increased lactate production compared with drug treatment or hypoxia alone. We conclude that pathogenic factors that interact to produce halothane hepatotoxicity act directly and jointly on isolated liver cells to produce energy deficits within 10 min. Differences in the relative importance of pathogenic factors in vitro and in vivo suggest that short-term, direct effects on hepatocellular energy status are not solely responsible for halothane hepatotoxicity.

pubmed23n0125_13721

Cardiovascular effects of and interaction between calcium blocking drugs and anesthetics in chronically instrumented dogs. III. Nicardipine and isoflurane.

To assess the interaction between isoflurane and the new calcium channel blocker, nicardipine, mongrel dogs were chronically instrumented to allow the following measurements: aortic, left ventricular and left atrial pressures; heart rate; cardiac output; and carotid, coronary, and renal blood flows. The hemodynamic effects of intravenous nicardipine 5, 10, 30, and 50 micrograms/kg were measured in awake dogs and during 1.6 and 3.0 per cent (end-tidal) isoflurane anesthesia. Nicardipine induced a dose-dependent fall in mean arterial pressure in both awake dogs and during 1.6 and 3.0 per cent isoflurane anesthesia. Heart rate and cardiac output were increased in proportion to the nicardipine dose in the awake dogs and, to a lesser degree, in the dogs anesthetized with 1.6 per cent isoflurane, but did not change during 3.0 per cent isoflurane anesthesia. Left atrial pressure was unchanged by nicardipine in awake dogs and during anesthesia. Left ventricular maximum rate of tension development (dP/dt) increased in awake dogs and decreased during anesthesia. Coronary blood flow increased dose dependently without anesthesia, and, to a smaller degree, during anesthesia. Nicardipine increased carotid blood flow without anesthesia, whereas it was unchanged during anesthesia. Renal blood flow was unchanged in awake dogs and decreased during anesthesia. The authors conclude that nicardipine is a potent vasodilator that minimally affects cardiac function and regional blood flow in the presence of isoflurane. The interactions between nicardipine and isoflurane are mainly the result of the isoflurane-induced inhibition of the reflex tachycardia elicited by nicardipine.

Cardiovascular effects of and interaction between calcium blocking drugs and anesthetics in chronically instrumented dogs. III. Nicardipine and isoflurane. To assess the interaction between isoflurane and the new calcium channel blocker, nicardipine, mongrel dogs were chronically instrumented to allow the following measurements: aortic, left ventricular and left atrial pressures; heart rate; cardiac output; and carotid, coronary, and renal blood flows. The hemodynamic effects of intravenous nicardipine 5, 10, 30, and 50 micrograms/kg were measured in awake dogs and during 1.6 and 3.0 per cent (end-tidal) isoflurane anesthesia. Nicardipine induced a dose-dependent fall in mean arterial pressure in both awake dogs and during 1.6 and 3.0 per cent isoflurane anesthesia. Heart rate and cardiac output were increased in proportion to the nicardipine dose in the awake dogs and, to a lesser degree, in the dogs anesthetized with 1.6 per cent isoflurane, but did not change during 3.0 per cent isoflurane anesthesia. Left atrial pressure was unchanged by nicardipine in awake dogs and during anesthesia. Left ventricular maximum rate of tension development (dP/dt) increased in awake dogs and decreased during anesthesia. Coronary blood flow increased dose dependently without anesthesia, and, to a smaller degree, during anesthesia. Nicardipine increased carotid blood flow without anesthesia, whereas it was unchanged during anesthesia. Renal blood flow was unchanged in awake dogs and decreased during anesthesia. The authors conclude that nicardipine is a potent vasodilator that minimally affects cardiac function and regional blood flow in the presence of isoflurane. The interactions between nicardipine and isoflurane are mainly the result of the isoflurane-induced inhibition of the reflex tachycardia elicited by nicardipine.

pubmed23n0125_13722

Noninvasive evaluation of breathing pattern and thoraco-abdominal motion following the infusion of ketamine or droperidol in humans.

The authors compared the respiratory effects of an intravenous infusion of ketamine (1 mg X kg-1) with droperidol (0.1 mg X kg-1), or placebo on three different occasions in a double-blind, randomized fashion in eight healthy volunteers. Breathing pattern, thoraco-abdominal motion, end-expiratory positions of the rib cage and abdomen, arterial hemoglobin oxygen saturation (SaO2), and end-tidal carbon dioxide concentration (FECO2) were continuously measured with noninvasive techniques. During the 1-h monitoring period following drug injection, droperidol produced occasionally significant but clinically unimportant differences in respiratory variables when compared with placebo. In contrast, ketamine induced a significant (P less than 0.001) and persistent increase in minute ventilation (+75%) from 5 to 20 min after start of infusion by increasing both the driving (i.e., tidal volume/inspiratory time [VT/Ti]) and the timing (i.e., inspiratory time/total respiratory cycle time [Ti/Ttot]) components of ventilation (Milic-Emili J, Grunstein MM: Chest 70 (Suppl): 131-133, 1976). This was obtained without any significant change in end-expiratory positions or change in relative rib cage contribution to tidal volume. Despite multiple apneic episodes observed with ketamine, the subjects maintained a stable SaO2 and FECO2, indicating no resting respiratory depression. This study, performed with a noninvasive respiratory monitoring technique, confirms that droperidol infused over 5 min at a clinically used dosage does not cause respiratory depression in healthy subjects, whereas ketamine produces an important ventilatory stimulation.

Noninvasive evaluation of breathing pattern and thoraco-abdominal motion following the infusion of ketamine or droperidol in humans. The authors compared the respiratory effects of an intravenous infusion of ketamine (1 mg X kg-1) with droperidol (0.1 mg X kg-1), or placebo on three different occasions in a double-blind, randomized fashion in eight healthy volunteers. Breathing pattern, thoraco-abdominal motion, end-expiratory positions of the rib cage and abdomen, arterial hemoglobin oxygen saturation (SaO2), and end-tidal carbon dioxide concentration (FECO2) were continuously measured with noninvasive techniques. During the 1-h monitoring period following drug injection, droperidol produced occasionally significant but clinically unimportant differences in respiratory variables when compared with placebo. In contrast, ketamine induced a significant (P less than 0.001) and persistent increase in minute ventilation (+75%) from 5 to 20 min after start of infusion by increasing both the driving (i.e., tidal volume/inspiratory time [VT/Ti]) and the timing (i.e., inspiratory time/total respiratory cycle time [Ti/Ttot]) components of ventilation (Milic-Emili J, Grunstein MM: Chest 70 (Suppl): 131-133, 1976). This was obtained without any significant change in end-expiratory positions or change in relative rib cage contribution to tidal volume. Despite multiple apneic episodes observed with ketamine, the subjects maintained a stable SaO2 and FECO2, indicating no resting respiratory depression. This study, performed with a noninvasive respiratory monitoring technique, confirms that droperidol infused over 5 min at a clinically used dosage does not cause respiratory depression in healthy subjects, whereas ketamine produces an important ventilatory stimulation.

pubmed23n0125_13723

Continuous-flow apneic ventilation during thoracotomy.

Continuous-flow apneic ventilation (CFAV) by endobronchial insufflation of conditioned gas was evaluated in dogs during thoracotomy. In Group 1 (n = 6), dogs were anesthetized with pentobarbital (25 mg/kg). An endobronchial catheter (2.5 mm ID) was introduced into each mainstem bronchus using a fiberoptic bronchoscope and held in place by an endotracheal tube. Before the onset of CFAV (total flow 1.01 X kg-1 X min-1, the animals were paralyzed with pancuronium bromide and muscle relaxation was monitored with a peripheral nerve stimulator. The CFAV delivery system consisted of a flow meter, air/oxygen blender, oxygen analyzer, heated humidifier, and ultrasonic spirometer. Blood gas values were measured after 30 min of spontaneous ventilation, and CFAV with: 1) closed chest, fractional inspired O2 concentration (FIO2) 0.21; 2) open chest, FIO2 0.21; 3) open chest, FIO2 0.21, continuous positive airway pressure (CPAP) 5 mmHg; and 4) open chest FIO2 0.4, CPAP 5 mmHg. This last combination resulted in a mean PaO2 of 113.1 +/- 5.5 (SEM) mmHg and a PaCO2 of 35.0 +/- 2.1 (SEM) mmHg. In Group 2 (n = 6), animals with open chests were ventilated with CFAV (FIO2 0.4 and CPAP 5 mmHg) for 5 h. Adequate oxygenation and ventilation were achieved. PaCO2 after 5 h of CFAV was 41.8 +/- 1.9 (SEM) mmHg compared with 40.8 +/- 1.9 (SEM) mmHg during spontaneous breathing. PaO2 after 5 h of CFAV was 138.1 +/- 11.7 (SEM) mmHg. There were no significant changes observed in vascular pressures. Significant differences in other hemodynamic parameters were probably due to pentobarbital anesthesia. Adequate gas exchange can be achieved during CFAV in dogs with open chests for 5 h.

Continuous-flow apneic ventilation during thoracotomy. Continuous-flow apneic ventilation (CFAV) by endobronchial insufflation of conditioned gas was evaluated in dogs during thoracotomy. In Group 1 (n = 6), dogs were anesthetized with pentobarbital (25 mg/kg). An endobronchial catheter (2.5 mm ID) was introduced into each mainstem bronchus using a fiberoptic bronchoscope and held in place by an endotracheal tube. Before the onset of CFAV (total flow 1.01 X kg-1 X min-1, the animals were paralyzed with pancuronium bromide and muscle relaxation was monitored with a peripheral nerve stimulator. The CFAV delivery system consisted of a flow meter, air/oxygen blender, oxygen analyzer, heated humidifier, and ultrasonic spirometer. Blood gas values were measured after 30 min of spontaneous ventilation, and CFAV with: 1) closed chest, fractional inspired O2 concentration (FIO2) 0.21; 2) open chest, FIO2 0.21; 3) open chest, FIO2 0.21, continuous positive airway pressure (CPAP) 5 mmHg; and 4) open chest FIO2 0.4, CPAP 5 mmHg. This last combination resulted in a mean PaO2 of 113.1 +/- 5.5 (SEM) mmHg and a PaCO2 of 35.0 +/- 2.1 (SEM) mmHg. In Group 2 (n = 6), animals with open chests were ventilated with CFAV (FIO2 0.4 and CPAP 5 mmHg) for 5 h. Adequate oxygenation and ventilation were achieved. PaCO2 after 5 h of CFAV was 41.8 +/- 1.9 (SEM) mmHg compared with 40.8 +/- 1.9 (SEM) mmHg during spontaneous breathing. PaO2 after 5 h of CFAV was 138.1 +/- 11.7 (SEM) mmHg. There were no significant changes observed in vascular pressures. Significant differences in other hemodynamic parameters were probably due to pentobarbital anesthesia. Adequate gas exchange can be achieved during CFAV in dogs with open chests for 5 h.

pubmed23n0125_13724

Succinylcholine: mechanism of fasciculations and their prevention by d-tubocurarine or diphenylhydantoin.

Administration of d-tubocurarine (dTC) or diphenylhydantoin (DPH) was evaluated as a pretreatment to prevent succinylcholine (Sch) evoked fasciculations. Experiments were designed to determine the nature of the drug-drug interactions, sites of interaction, and site of fasciculation suppression. Sch is known to evoke repetitive discharge generation by motor nerve terminals (MNTs). Transmission of these prejunctional discharges causes fasciculations. A cat soleus neuromuscular preparation in situ, which enables recording of nerve action potentials initiated by MNTs, their transmitted muscle action potentials, and the resultant contractile responses, was used to explore Sch effects before and after iv pretreatment with dTC or DPH. dTC is known to act prejunctionally to suppress repetitive discharges initiated by facilitatory drugs and tetanic conditioning of MNTs. Accordingly, pretreatment with dTC 50 micrograms X kg-1 suppressed the Sch-induced MNT repetitive discharging and correspondingly suppressed generalized fasciculations without affecting twitch. This dTC dose, however, also reduced Sch blocking potency by 33%, slowed its rate, and shortened block duration. These latter effects represent competitive postjunctional antagonism. DPH is also known to suppress MNT repetitive discharging. Correspondingly, Sch-induced repetitive firing and ensuing fasciculations were suppressed by DPH (30 mg X kg-1) without affecting twitch. Unlike dTC, this DPH dose increased Sch blocking potency by 50%, increased the initial rate of block, and did not alter block duration. These DPH effects were dose-dependent and within the anticonvulsant range for cats. Therefore, patients with anticonvulsant levels of DPH may not require pretreatment before Sch.(ABSTRACT TRUNCATED AT 250 WORDS)

Succinylcholine: mechanism of fasciculations and their prevention by d-tubocurarine or diphenylhydantoin. Administration of d-tubocurarine (dTC) or diphenylhydantoin (DPH) was evaluated as a pretreatment to prevent succinylcholine (Sch) evoked fasciculations. Experiments were designed to determine the nature of the drug-drug interactions, sites of interaction, and site of fasciculation suppression. Sch is known to evoke repetitive discharge generation by motor nerve terminals (MNTs). Transmission of these prejunctional discharges causes fasciculations. A cat soleus neuromuscular preparation in situ, which enables recording of nerve action potentials initiated by MNTs, their transmitted muscle action potentials, and the resultant contractile responses, was used to explore Sch effects before and after iv pretreatment with dTC or DPH. dTC is known to act prejunctionally to suppress repetitive discharges initiated by facilitatory drugs and tetanic conditioning of MNTs. Accordingly, pretreatment with dTC 50 micrograms X kg-1 suppressed the Sch-induced MNT repetitive discharging and correspondingly suppressed generalized fasciculations without affecting twitch. This dTC dose, however, also reduced Sch blocking potency by 33%, slowed its rate, and shortened block duration. These latter effects represent competitive postjunctional antagonism. DPH is also known to suppress MNT repetitive discharging. Correspondingly, Sch-induced repetitive firing and ensuing fasciculations were suppressed by DPH (30 mg X kg-1) without affecting twitch. Unlike dTC, this DPH dose increased Sch blocking potency by 50%, increased the initial rate of block, and did not alter block duration. These DPH effects were dose-dependent and within the anticonvulsant range for cats. Therefore, patients with anticonvulsant levels of DPH may not require pretreatment before Sch.(ABSTRACT TRUNCATED AT 250 WORDS)

pubmed23n0125_13725

A departmental policy addressing chemical substance abuse.

Substance abuse is a major socioeconomic problem. However, the ready availability of potent narcotic and sedative drugs probably constitutes a unique risk for anesthesiologists. Until recently, few anesthesia departments were prepared to recognize or safely manage afflicted colleagues. Because we felt it important to educate our staff and residents and to have a response mechanism established prior to the advent of a substance abuse problem, a departmental committee was formed to develop a Substance Abuse Policy. The policy has served to increase our general awareness and to direct our actions effectively when dealing with physician impairment. It is presented here in the belief that other departments might find it useful in tailoring their approach to this problem.

A departmental policy addressing chemical substance abuse. Substance abuse is a major socioeconomic problem. However, the ready availability of potent narcotic and sedative drugs probably constitutes a unique risk for anesthesiologists. Until recently, few anesthesia departments were prepared to recognize or safely manage afflicted colleagues. Because we felt it important to educate our staff and residents and to have a response mechanism established prior to the advent of a substance abuse problem, a departmental committee was formed to develop a Substance Abuse Policy. The policy has served to increase our general awareness and to direct our actions effectively when dealing with physician impairment. It is presented here in the belief that other departments might find it useful in tailoring their approach to this problem.

pubmed23n0125_13726

Validation of quantitative intraoperative transesophageal echocardiography.

Transesophageal echocardiography (TEE) is a new monitoring technique that images the heart and provides information on regional wall motion and left ventricular filling. However, despite its potential for inaccuracy due to its retrocardiac position and angulation, TEE has not been validated by another imaging technique. Using direct on-heart echocardiography (OHE) as a standard, the authors evaluated the ability of TEE to measure accurately left ventricular end-diastolic area (EDa), end-systolic area (ESa), and ejection fraction area (EFa). Ten patients with coronary artery disease without evidence of valvular dysfunction undergoing myocardial revascularization were studied. A Diasonics 3.5 MHz two-dimensional TEE probe was introduced into each patient's esophagus and positioned to obtain a view equivalent to the parasternal short-axis projection. A similar view was obtained by OHE using a sterilely prepared 3 MHz ATL probe placed on either the pericardium or epicardium. In each patient, immediately prior to and after pericardiotomy, both transesophageal and on-heart short-axis views at the level of the papillary muscles were obtained. Using a dedicated Diasonics computer echoanalyzer, EDa and ESa from four consecutive cardiac cycles were outlined with a light pen and averaged. EFa was calculated by the formula EFa = (EDa - ESa)/EDa. Seventeen comparable transesophageal and on-heart echocardiograms were obtained. ESa by TEE correlated well with ESa by OHE (15.13 +/- 9.62 cm2 vs. 14.92 +/- 10.53 cm2; r = 0.94). Similar results were obtained for EDa (27.75 +/- 9.88 cm2 vs. 30.40 +/- 13.99 cm2; r = 0.88) and EFa (0.49 +/- 0.17 vs. 0.54 +/- 0.13; r = 0.92). filling and ejection.

Validation of quantitative intraoperative transesophageal echocardiography. Transesophageal echocardiography (TEE) is a new monitoring technique that images the heart and provides information on regional wall motion and left ventricular filling. However, despite its potential for inaccuracy due to its retrocardiac position and angulation, TEE has not been validated by another imaging technique. Using direct on-heart echocardiography (OHE) as a standard, the authors evaluated the ability of TEE to measure accurately left ventricular end-diastolic area (EDa), end-systolic area (ESa), and ejection fraction area (EFa). Ten patients with coronary artery disease without evidence of valvular dysfunction undergoing myocardial revascularization were studied. A Diasonics 3.5 MHz two-dimensional TEE probe was introduced into each patient's esophagus and positioned to obtain a view equivalent to the parasternal short-axis projection. A similar view was obtained by OHE using a sterilely prepared 3 MHz ATL probe placed on either the pericardium or epicardium. In each patient, immediately prior to and after pericardiotomy, both transesophageal and on-heart short-axis views at the level of the papillary muscles were obtained. Using a dedicated Diasonics computer echoanalyzer, EDa and ESa from four consecutive cardiac cycles were outlined with a light pen and averaged. EFa was calculated by the formula EFa = (EDa - ESa)/EDa. Seventeen comparable transesophageal and on-heart echocardiograms were obtained. ESa by TEE correlated well with ESa by OHE (15.13 +/- 9.62 cm2 vs. 14.92 +/- 10.53 cm2; r = 0.94). Similar results were obtained for EDa (27.75 +/- 9.88 cm2 vs. 30.40 +/- 13.99 cm2; r = 0.88) and EFa (0.49 +/- 0.17 vs. 0.54 +/- 0.13; r = 0.92). filling and ejection.

pubmed23n0125_13727

Effect of normal and preeclamptic pregnancies on the oxyhemoglobin dissociation curve.

Hemoglobin affinity for oxygen in whole blood of ten normal nonpregnant women, ten normal pregnant women at first trimester, ten normal pregnant women at second trimester, 24 normal pregnant women at or near term, and 14 pregnant women with preeclampsia at or near term was studied. The mean P-50 values for normal nonpregnant women, normal pregnant women in first trimester, second trimester, and at or near term were 26.7 +/- 0.11 mmHg, 27.8 +/- 0.08 mmHg, 28.8 +/- 0.17 mmHg, and 30.4 +/- 0.20 mmHg, respectively. The mean P-50 of pregnant women with preeclampsia at or near term was 25.1 +/- 0.38 mmHg. It is concluded that in normal pregnant women there was a significant shift of P-50 to the right compared with the normal nonpregnant women (P less than 0.01), and the extent of this shift to the right is directly related to the duration of the pregnancy. In addition, preeclamptic parturients showed a significant shift of P-50 to the left when compared with normal pregnant women at or near term (P less than 0.001).

Effect of normal and preeclamptic pregnancies on the oxyhemoglobin dissociation curve. Hemoglobin affinity for oxygen in whole blood of ten normal nonpregnant women, ten normal pregnant women at first trimester, ten normal pregnant women at second trimester, 24 normal pregnant women at or near term, and 14 pregnant women with preeclampsia at or near term was studied. The mean P-50 values for normal nonpregnant women, normal pregnant women in first trimester, second trimester, and at or near term were 26.7 +/- 0.11 mmHg, 27.8 +/- 0.08 mmHg, 28.8 +/- 0.17 mmHg, and 30.4 +/- 0.20 mmHg, respectively. The mean P-50 of pregnant women with preeclampsia at or near term was 25.1 +/- 0.38 mmHg. It is concluded that in normal pregnant women there was a significant shift of P-50 to the right compared with the normal nonpregnant women (P less than 0.01), and the extent of this shift to the right is directly related to the duration of the pregnancy. In addition, preeclamptic parturients showed a significant shift of P-50 to the left when compared with normal pregnant women at or near term (P less than 0.001).

pubmed23n0125_13728

Bedside hemodynamic monitoring in the management of acute cardiac patients.

Hemodynamic monitoring was performed in 100 acutely ill patients admitted to a Coronary Care Unit, 72 of whom had sustained an acute myocardial infarction (AMI). In patients with AMI, the initial pulmonary capillary wedge (PCW) pressure was related to prognosis, with a mortality of 68% in those with elevated pressures. The mean PCW pressure was 25 mmHg in those who died as compared with 17 mmHg (p less than 0.01) in the survivors. Cardiac catheterization confirmed the clinical impression of high risk in the majority of cases but also identified those in whom the clinical signs were misleading; 9 of those with AMI (12.5%) had high PCW pressures in the absence of clinical or radiological heart failure. The diagnosis of serious hemodynamic complications of AMI (right ventricular infarction, ventricular septal defect, and mitral regurgitation) was established in 14 cases (19%). There were no serious complications related to catheterization, and we conclude that pulmonary artery catheterization is safe and of clinical value in acutely ill cardiac patients.

Bedside hemodynamic monitoring in the management of acute cardiac patients. Hemodynamic monitoring was performed in 100 acutely ill patients admitted to a Coronary Care Unit, 72 of whom had sustained an acute myocardial infarction (AMI). In patients with AMI, the initial pulmonary capillary wedge (PCW) pressure was related to prognosis, with a mortality of 68% in those with elevated pressures. The mean PCW pressure was 25 mmHg in those who died as compared with 17 mmHg (p less than 0.01) in the survivors. Cardiac catheterization confirmed the clinical impression of high risk in the majority of cases but also identified those in whom the clinical signs were misleading; 9 of those with AMI (12.5%) had high PCW pressures in the absence of clinical or radiological heart failure. The diagnosis of serious hemodynamic complications of AMI (right ventricular infarction, ventricular septal defect, and mitral regurgitation) was established in 14 cases (19%). There were no serious complications related to catheterization, and we conclude that pulmonary artery catheterization is safe and of clinical value in acutely ill cardiac patients.

pubmed23n0125_13729

Cardiovascular effects of polycyclic antidepressants.

Every day, thousands of people in the United States receive polycyclic antidepressants (PCAs) prescribed by their primary care physicians. We have examined the cardiovascular manifestations of polycyclic antidepressants in 14 patients with primary depression receiving various antidepressant drugs for a period of six months or more. None had suffered from any systemic illness. All 14 received a commonly prescribed PCA, such as doxepin, amitriptyline, imipramine, desipramine or amoxapine, in therapeutic doses. None developed any significant adverse reaction. Left ventricular function, as determined by M-mode echocardiogram, was within normal limit in all patients. A twenty-four-hour ambulatory electrocardiogram revealed rare supraventricular ectopic beats in 8 patients and ventricular ectopic activity (Lown grade 1) in 4 patients. In 1 patient, who received additional lithium carbonate for treatment of depression, both supraventricular and ventricular ectopic activity developed on rare occasions. In 4 of the 8 patients the plasma concentration of PCA was optimal or near optimal, and in 4 of the 8 patients it was subtherapeutic (even though these 4 patients had shown clinical improvement with continued polycyclic therapy). It appears from the present data that antidepressant therapy with PCAs prescribed in therapeutic doses was well tolerated, without any significant adverse cardiovascular reaction, in otherwise healthy young patients with primary depression.

Cardiovascular effects of polycyclic antidepressants. Every day, thousands of people in the United States receive polycyclic antidepressants (PCAs) prescribed by their primary care physicians. We have examined the cardiovascular manifestations of polycyclic antidepressants in 14 patients with primary depression receiving various antidepressant drugs for a period of six months or more. None had suffered from any systemic illness. All 14 received a commonly prescribed PCA, such as doxepin, amitriptyline, imipramine, desipramine or amoxapine, in therapeutic doses. None developed any significant adverse reaction. Left ventricular function, as determined by M-mode echocardiogram, was within normal limit in all patients. A twenty-four-hour ambulatory electrocardiogram revealed rare supraventricular ectopic beats in 8 patients and ventricular ectopic activity (Lown grade 1) in 4 patients. In 1 patient, who received additional lithium carbonate for treatment of depression, both supraventricular and ventricular ectopic activity developed on rare occasions. In 4 of the 8 patients the plasma concentration of PCA was optimal or near optimal, and in 4 of the 8 patients it was subtherapeutic (even though these 4 patients had shown clinical improvement with continued polycyclic therapy). It appears from the present data that antidepressant therapy with PCAs prescribed in therapeutic doses was well tolerated, without any significant adverse cardiovascular reaction, in otherwise healthy young patients with primary depression.

pubmed23n0125_13730

Tissue fibrinolytic activity in different types of varicose veins.

The fibrinolytic activity of the venous wall was investigated by using Todd's technique in 92 patients with different types of varicosis. A control group consisted of 19 patients with apparently normal superficial veins who had had a saphenectomy prior to an aortocoronary bypass operation. Fibrinolytic activity was mainly localized in the adventitia of varicose and normal veins. It significantly decreased in the distal regions of all types of varicosis. The highest fibrinolytic activity was detected in the proximal part of the varicose vena saphena magna and the lowest values were observed in the perforating and side branch veins of the calf. Fibrinolytic activity is higher (p less than 0.077) in the normal vena saphena magna than in the varicose vena saphena of the calf. Older patients show a loss of fibrinolytic activity in their vena saphena magna. Obese patients have less fibrinolytic activity in varicose calf veins than patients with normal weight do.

Tissue fibrinolytic activity in different types of varicose veins. The fibrinolytic activity of the venous wall was investigated by using Todd's technique in 92 patients with different types of varicosis. A control group consisted of 19 patients with apparently normal superficial veins who had had a saphenectomy prior to an aortocoronary bypass operation. Fibrinolytic activity was mainly localized in the adventitia of varicose and normal veins. It significantly decreased in the distal regions of all types of varicosis. The highest fibrinolytic activity was detected in the proximal part of the varicose vena saphena magna and the lowest values were observed in the perforating and side branch veins of the calf. Fibrinolytic activity is higher (p less than 0.077) in the normal vena saphena magna than in the varicose vena saphena of the calf. Older patients show a loss of fibrinolytic activity in their vena saphena magna. Obese patients have less fibrinolytic activity in varicose calf veins than patients with normal weight do.

pubmed23n0125_13731

Cerebral hemorrhagic infarction associated with anabolic steroid therapy for hypoplastic anemia.

A twenty-two-year-old student with hypoplastic anemia was treated with large doses of anabolic steroid hormone preparations for two months. Cerebral hemorrhagic infarction shown by CAT scan developed when his hematologic conditions were improving. In treating hypoplastic anemia with anabolic steroid hormone preparations, one should be aware of the possible development of cerebral hemorrhagic infarction at the time of hematologic improvement.

Cerebral hemorrhagic infarction associated with anabolic steroid therapy for hypoplastic anemia. A twenty-two-year-old student with hypoplastic anemia was treated with large doses of anabolic steroid hormone preparations for two months. Cerebral hemorrhagic infarction shown by CAT scan developed when his hematologic conditions were improving. In treating hypoplastic anemia with anabolic steroid hormone preparations, one should be aware of the possible development of cerebral hemorrhagic infarction at the time of hematologic improvement.

pubmed23n0125_13732

Vertebral arteriovenous fistula following central venous cannulation: a case report.

The authors report a case of vertebral arteriovenous fistula that has been disclosed three years after central venous cannulation (CVC). The real incidence of this complication is discussed and various clinical presentations are enumerated. From a review of the literature, some recommendations are made to prevent the diagnosis from being missed and chiefly to reduce the risk of arterial puncture that results in fistula formation.

Vertebral arteriovenous fistula following central venous cannulation: a case report. The authors report a case of vertebral arteriovenous fistula that has been disclosed three years after central venous cannulation (CVC). The real incidence of this complication is discussed and various clinical presentations are enumerated. From a review of the literature, some recommendations are made to prevent the diagnosis from being missed and chiefly to reduce the risk of arterial puncture that results in fistula formation.

pubmed23n0125_13733

Aortic connective tissue in atherosclerotic aorta--a biochemical study.

Biochemical analysis of the extracellular matrix of human aortas was performed on samples of ascending and descending aortas affected by atherosclerosis in comparison with a control group of nonatherosclerotic aortas. Ulcerated or heavily calcified atheromas were excised and excluded from the analysis in order to differentiate biochemical alterations leading to the formation of atheromas from those due to complications of already formed atheromas. Our results show that the development of atheromas brings about an extensive destruction of elastic fibers and muscular cells, and their place is occupied by other components of the extracellular matrix, most notably, collagen, non-uronic sugars, water, and lipids, which were found significantly increased.

Aortic connective tissue in atherosclerotic aorta--a biochemical study. Biochemical analysis of the extracellular matrix of human aortas was performed on samples of ascending and descending aortas affected by atherosclerosis in comparison with a control group of nonatherosclerotic aortas. Ulcerated or heavily calcified atheromas were excised and excluded from the analysis in order to differentiate biochemical alterations leading to the formation of atheromas from those due to complications of already formed atheromas. Our results show that the development of atheromas brings about an extensive destruction of elastic fibers and muscular cells, and their place is occupied by other components of the extracellular matrix, most notably, collagen, non-uronic sugars, water, and lipids, which were found significantly increased.

pubmed23n0125_13734

Autopsy incidence of pulmonary vascular episodes. A study of 218 cases.

Pulmonary thromboembolism is a rarity in India. This common clinical impression has so far not been tested. Among 7000 autopsies between 1964 and 1980, a total of 218 cases (126 males and 92 females) were recorded to have thrombosis and/or embolism and/or infarction in the lungs. This incidence of 3.1% is far lower than that reported in the West and similar to the low incidence in Africa. Of the 218 cases, 42.6% had a cardiac disease, 18.3% had systemic septicemia, 13% had a malignancy, 12.8% had pulmonary disease, and the remaining suffered from diseases of liver, kidney, CNS, etc. Of the 218 cases, 141 (64.6%) showed only infarcts, 40 (18.3%) had only thromboemboli, and 37 (16.9%) showed both events. In view of the overlap among these three conditions and their essential pathophysiologic identity (thrombus/embolism/infarction), it is suggested that these be grouped under the name "pulmonary vascular episode."

Autopsy incidence of pulmonary vascular episodes. A study of 218 cases. Pulmonary thromboembolism is a rarity in India. This common clinical impression has so far not been tested. Among 7000 autopsies between 1964 and 1980, a total of 218 cases (126 males and 92 females) were recorded to have thrombosis and/or embolism and/or infarction in the lungs. This incidence of 3.1% is far lower than that reported in the West and similar to the low incidence in Africa. Of the 218 cases, 42.6% had a cardiac disease, 18.3% had systemic septicemia, 13% had a malignancy, 12.8% had pulmonary disease, and the remaining suffered from diseases of liver, kidney, CNS, etc. Of the 218 cases, 141 (64.6%) showed only infarcts, 40 (18.3%) had only thromboemboli, and 37 (16.9%) showed both events. In view of the overlap among these three conditions and their essential pathophysiologic identity (thrombus/embolism/infarction), it is suggested that these be grouped under the name "pulmonary vascular episode."

pubmed23n0125_13735

Angina pectoris and acute myocardial infarction due to "slow-flow phenomenon" in nonatherosclerotic coronary arteries: a case report.

A 51 year-old White woman with angina pectoris and nonatheromatous coronary artery disease is presented. Cardiac catheterization demonstrated a "slow-flow phenomenon" in the left coronary artery accompanied by severe angina pectoris and anterolateral ST-segment elevation and culminating in an acute nontransmural myocardial infarction. At repeat coronary arteriography, ergonovine maleate provocation proved negative. This patient is unique, since the previously documented 6 cases with this coronary cineangiographic response did not exhibit angina pectoris or ECG evidence of myocardial ischemia during the "slow-flow phenomenon," and none was complicated by an acute myocardial infarction. Various aspects of the pathophysiology of angina pectoris in this patient, including the recently described "reduced vasodilator reserve" concept, are briefly outlined.

Angina pectoris and acute myocardial infarction due to "slow-flow phenomenon" in nonatherosclerotic coronary arteries: a case report. A 51 year-old White woman with angina pectoris and nonatheromatous coronary artery disease is presented. Cardiac catheterization demonstrated a "slow-flow phenomenon" in the left coronary artery accompanied by severe angina pectoris and anterolateral ST-segment elevation and culminating in an acute nontransmural myocardial infarction. At repeat coronary arteriography, ergonovine maleate provocation proved negative. This patient is unique, since the previously documented 6 cases with this coronary cineangiographic response did not exhibit angina pectoris or ECG evidence of myocardial ischemia during the "slow-flow phenomenon," and none was complicated by an acute myocardial infarction. Various aspects of the pathophysiology of angina pectoris in this patient, including the recently described "reduced vasodilator reserve" concept, are briefly outlined.

pubmed23n0125_13736

Catheter separation during cardiac catheterization and coronary angiography. A report of four incidents and review of the literature.

Four incidents of catheter separation in three patients during left heart catheterization and coronary angiography are presented. In two cases the catheters had been used before. After the second incident, we decided to use angiography catheters once only. All catheter fragments were removed under local anesthesia, and no further complications arose. In all incidents the point of catheter fracture was near the bond between the body of the catheter and its tip. We believe that catheter separation is related to polymer aging. The catheters in the third and fourth incidents were unused, but there was no expiration date on their packages. We recommend that expiration dates be clearly stated on every catheter package, catheter manufactures issue warnings not to use catheters after expiration date, and unused angiography catheters with no expiration date not be used. We also suggest that consideration be given to using catheters once only.

Catheter separation during cardiac catheterization and coronary angiography. A report of four incidents and review of the literature. Four incidents of catheter separation in three patients during left heart catheterization and coronary angiography are presented. In two cases the catheters had been used before. After the second incident, we decided to use angiography catheters once only. All catheter fragments were removed under local anesthesia, and no further complications arose. In all incidents the point of catheter fracture was near the bond between the body of the catheter and its tip. We believe that catheter separation is related to polymer aging. The catheters in the third and fourth incidents were unused, but there was no expiration date on their packages. We recommend that expiration dates be clearly stated on every catheter package, catheter manufactures issue warnings not to use catheters after expiration date, and unused angiography catheters with no expiration date not be used. We also suggest that consideration be given to using catheters once only.

pubmed23n0125_13737

Graphic representation of carotid artery bruits.

Auscultation of the neck for detection of bruits over the carotid arteries has become a necessary part of any complete physical examination. Early detection of atherosclerotic occlusive disease in these vessels can play a major role in prevention of stroke. Such findings have previously been recorded as word descriptions in varying degrees of detail. A method is presented for quickly and easily recording the presence and character of cervical bruits, plus additional important information, including blood pressure, pulse, heart rhythm, prior vascular surgery, and known occlusion in these vessels. The figure used allows others to rapidly perceive this information and enables ready comparison on successive examinations.

Graphic representation of carotid artery bruits. Auscultation of the neck for detection of bruits over the carotid arteries has become a necessary part of any complete physical examination. Early detection of atherosclerotic occlusive disease in these vessels can play a major role in prevention of stroke. Such findings have previously been recorded as word descriptions in varying degrees of detail. A method is presented for quickly and easily recording the presence and character of cervical bruits, plus additional important information, including blood pressure, pulse, heart rhythm, prior vascular surgery, and known occlusion in these vessels. The figure used allows others to rapidly perceive this information and enables ready comparison on successive examinations.

pubmed23n0125_13738

Doppler color flow mapping of the fetal heart.

The recently developed innovative technique of Doppler color flow mapping permits real time visualization of blood flow patterns superimposed on two-dimensional echocardiographic images of the heart. This study demonstrates, for the first time, the usefulness of this technique in characterizing fetal cardiac dynamics.

Doppler color flow mapping of the fetal heart. The recently developed innovative technique of Doppler color flow mapping permits real time visualization of blood flow patterns superimposed on two-dimensional echocardiographic images of the heart. This study demonstrates, for the first time, the usefulness of this technique in characterizing fetal cardiac dynamics.

pubmed23n0125_13739

Central inhibition of sympathetic overdrive by clonidine in acute myocardial infarction with systolic hypertension. Haemodynamic study.

Intravenous clonidine was used to treat systolic hypertension (systolic blood pressure greater than 160 mm Hg) in 15 patients with acute myocardial infarction and documented sympathetic overactivity (high plasma norepinephrine). Its effects on haemodynamics and blood gases were studied. After one hour, clonidine significantly reduced the systolic (195 +/- 7 to 137 +/- 7 mm Hg, p less than 0.01) and diastolic (81 +/- 4 to 60 +/- 3 mm Hg, p less than 0.01) blood pressures as well as the systemic vascular resistance (26 +/- 2 to 20 +/- 1 IU, p less than 0.01). The cardiac index was reduced from 2.8 +/- 0.2 to 2.4 +/- 0.2 l/min X m2, p less than 0.01. This change was related to a reduction of the heart rate (92 +/- 4 to 81 +/- 4 beats/min, p less than 0.01) as the stroke index was unchanged. Pulmonary wedge pressure (15 +/- 3 to 10 +/- 2 mm Hg, p less than 0.01) and rate pressure product (18.034 +/- 1.159 to 11.274 +/- 917 mm Hg, beats/min, p less than 0.01) were also significantly decreased. The arterial oxygen tension did not change significantly but there was a significant drop in the mixed venous oxygen saturation (63 +/- 2 to 61 +/- 2%, p less than 0.02) and oxygen transport (433 +/- 41 to 409 +/- 36, p less than 0.01). Clonidine is thus able to normalize blood pressure in acute myocardial infarction; this is accompanied by a reduction in myocardial oxygen requirements and pulmonary wedge pressure. Oxygen transport to the tissues, however, may be decreased.

Central inhibition of sympathetic overdrive by clonidine in acute myocardial infarction with systolic hypertension. Haemodynamic study. Intravenous clonidine was used to treat systolic hypertension (systolic blood pressure greater than 160 mm Hg) in 15 patients with acute myocardial infarction and documented sympathetic overactivity (high plasma norepinephrine). Its effects on haemodynamics and blood gases were studied. After one hour, clonidine significantly reduced the systolic (195 +/- 7 to 137 +/- 7 mm Hg, p less than 0.01) and diastolic (81 +/- 4 to 60 +/- 3 mm Hg, p less than 0.01) blood pressures as well as the systemic vascular resistance (26 +/- 2 to 20 +/- 1 IU, p less than 0.01). The cardiac index was reduced from 2.8 +/- 0.2 to 2.4 +/- 0.2 l/min X m2, p less than 0.01. This change was related to a reduction of the heart rate (92 +/- 4 to 81 +/- 4 beats/min, p less than 0.01) as the stroke index was unchanged. Pulmonary wedge pressure (15 +/- 3 to 10 +/- 2 mm Hg, p less than 0.01) and rate pressure product (18.034 +/- 1.159 to 11.274 +/- 917 mm Hg, beats/min, p less than 0.01) were also significantly decreased. The arterial oxygen tension did not change significantly but there was a significant drop in the mixed venous oxygen saturation (63 +/- 2 to 61 +/- 2%, p less than 0.02) and oxygen transport (433 +/- 41 to 409 +/- 36, p less than 0.01). Clonidine is thus able to normalize blood pressure in acute myocardial infarction; this is accompanied by a reduction in myocardial oxygen requirements and pulmonary wedge pressure. Oxygen transport to the tissues, however, may be decreased.

pubmed23n0125_13740

Mossy leg--an unusual therapeutic success.

Elephantiasis verrucosa nostrum occurs when lymphatic obstruction of sufficient magnitude causes back pressure in the lymphatic channels forceful enough to produce superficial vessel dilation which has a "pebbly" or cobble-stone appearance. Unchecked this can progress to a marked verrucous or mossy appearance, frequently complicated by infection, with swelling and gross deformity. The patient presented here was both post-phlebitic and post-lymphatic with significant deformity. He showed dramatic improvement on a regimen of antibiotics and elevation followed by decompression and routine use of pressure-gradient support garments. This has not recurred and he has remained under control since 1979.

Mossy leg--an unusual therapeutic success. Elephantiasis verrucosa nostrum occurs when lymphatic obstruction of sufficient magnitude causes back pressure in the lymphatic channels forceful enough to produce superficial vessel dilation which has a "pebbly" or cobble-stone appearance. Unchecked this can progress to a marked verrucous or mossy appearance, frequently complicated by infection, with swelling and gross deformity. The patient presented here was both post-phlebitic and post-lymphatic with significant deformity. He showed dramatic improvement on a regimen of antibiotics and elevation followed by decompression and routine use of pressure-gradient support garments. This has not recurred and he has remained under control since 1979.

pubmed23n0125_13741

tcpO2, arterial blood flow and lactate/pyruvate modifications induced by a single dose of naftidrofuryl IV during stage III lower-limb arteriopathies.

A single dose of 600 mg of naftidrofuryl perfused IV during three hours in 12 stage III arteriopathic patients did not modify the average blood flow at the calf but did significantly increase transcutaneous oxygen pressure (33 vs 52 mmHg) and also improved lactates (1.44 vs 0.88 mmol/liter), pyruvate (59 vs 79 mumol/liter), and lactates-pyruvates ratio (33.48 vs 20.92).

tcpO2, arterial blood flow and lactate/pyruvate modifications induced by a single dose of naftidrofuryl IV during stage III lower-limb arteriopathies. A single dose of 600 mg of naftidrofuryl perfused IV during three hours in 12 stage III arteriopathic patients did not modify the average blood flow at the calf but did significantly increase transcutaneous oxygen pressure (33 vs 52 mmHg) and also improved lactates (1.44 vs 0.88 mmol/liter), pyruvate (59 vs 79 mumol/liter), and lactates-pyruvates ratio (33.48 vs 20.92).

pubmed23n0125_13742

Muscular blood flow in patients with heart failure.

We studied reactive hyperemia in a group of patients with heart failure before and after therapy, since changes in the characteristics of muscular blood flow may influence the functional class of these patients. At the same time we evaluated some echocardiographic parameters too. When the patients improved clinically, they showed an increase in muscular blood flow at rest and in percent of fractional shortening and a decrease in peripheral vascular resistance. The reactive hyperemia did not change significantly. This fact probably depends on a maximal response to the postischemic hyperemia and represents the integrity of autoregulatory mechanisms.

Muscular blood flow in patients with heart failure. We studied reactive hyperemia in a group of patients with heart failure before and after therapy, since changes in the characteristics of muscular blood flow may influence the functional class of these patients. At the same time we evaluated some echocardiographic parameters too. When the patients improved clinically, they showed an increase in muscular blood flow at rest and in percent of fractional shortening and a decrease in peripheral vascular resistance. The reactive hyperemia did not change significantly. This fact probably depends on a maximal response to the postischemic hyperemia and represents the integrity of autoregulatory mechanisms.

pubmed23n0125_13743

Oral and dental complications of sickle cell disease in Nigerians.

A clinical evaluation of the oral and dental complications of sickle cell disease in Nigerians was carried out in 37 consecutive patients with homozygous sickle cell disease Hb-SS (Sicklers) compared to a control group of 24 persons with normal haemoglobin Hb-AA (control group) matched for age and sex. The significant abnormalities found in sicklers included intrinsic opacity of the teeth in 67.5% of sicklers compared to 28.83% in the control group; malocclusion of the teeth with over-jet and over-bite in 35% of sicklers compared to 16.66% in the control group; dental caries is present in 35.13% of sicklers which was less than its occurrence in 54% of the control group due to widespread avoidance of sweets by most local sicklers. Diastemata (gaps between the teeth) was present in approximately equal frequency in sicklers (27%) and control group (25%). In view of the aesthetic and medical implications of these abnormalities, it is recommended that sicklers should receive regular dental check-up with a view to ameliorating or preventing these complications by prophylactic measures including the use of orthodontic appliances such as braces, etc. The above findings are discussed in relation to the other complications of sickler cell disease in other organs of the body.

Oral and dental complications of sickle cell disease in Nigerians. A clinical evaluation of the oral and dental complications of sickle cell disease in Nigerians was carried out in 37 consecutive patients with homozygous sickle cell disease Hb-SS (Sicklers) compared to a control group of 24 persons with normal haemoglobin Hb-AA (control group) matched for age and sex. The significant abnormalities found in sicklers included intrinsic opacity of the teeth in 67.5% of sicklers compared to 28.83% in the control group; malocclusion of the teeth with over-jet and over-bite in 35% of sicklers compared to 16.66% in the control group; dental caries is present in 35.13% of sicklers which was less than its occurrence in 54% of the control group due to widespread avoidance of sweets by most local sicklers. Diastemata (gaps between the teeth) was present in approximately equal frequency in sicklers (27%) and control group (25%). In view of the aesthetic and medical implications of these abnormalities, it is recommended that sicklers should receive regular dental check-up with a view to ameliorating or preventing these complications by prophylactic measures including the use of orthodontic appliances such as braces, etc. The above findings are discussed in relation to the other complications of sickler cell disease in other organs of the body.

pubmed23n0125_13744

Circulating platelet aggregate size in ischemic heart disease.

Platelet aggregate size was measured in 178 patients with ischemic heart disease, among whom 56 had stable angina, 42 suffered from unstable angina, and 80 had had uncomplicated acute myocardial infarction. A group of 50 healthy volunteers and 20 hospitalized noncardiac patients served as controls. Venous blood (0.5 cc) was introduced into a solution containing 11.7 mM EDTA and 1.0 g formaldehyde. Platelet aggregate size was determined by microscopic reading as the number of platelets forming aggregates (per 1000 counted platelets) divided by the number of aggregates. Mean aggregate size was found not significantly different in both control groups, as well as in patients with stable angina and acute myocardial infarction (2.21 +/- 0.36 platelets, 2.20 +/- 0.58 platelets, 2.28 +/- 0.19 platelets, 2.76 +/- 1.07 platelets, respectively, p = NS). The highest value was found in the unstable angina group: 4.00 +/- 1.40 platelets (p less than 0.001 vs other studied groups). Platelet aggregate size was found not to be related to sex, age, medication, or coronary risk factors. Unstable angina may thus be a unique entity in ischemic heart disease concerning its platelet behavior, demonstrated in this study by the increased size of peripheral platelet aggregates, which may have pathogenetic, diagnostic, and eventual therapeutic implications.

Circulating platelet aggregate size in ischemic heart disease. Platelet aggregate size was measured in 178 patients with ischemic heart disease, among whom 56 had stable angina, 42 suffered from unstable angina, and 80 had had uncomplicated acute myocardial infarction. A group of 50 healthy volunteers and 20 hospitalized noncardiac patients served as controls. Venous blood (0.5 cc) was introduced into a solution containing 11.7 mM EDTA and 1.0 g formaldehyde. Platelet aggregate size was determined by microscopic reading as the number of platelets forming aggregates (per 1000 counted platelets) divided by the number of aggregates. Mean aggregate size was found not significantly different in both control groups, as well as in patients with stable angina and acute myocardial infarction (2.21 +/- 0.36 platelets, 2.20 +/- 0.58 platelets, 2.28 +/- 0.19 platelets, 2.76 +/- 1.07 platelets, respectively, p = NS). The highest value was found in the unstable angina group: 4.00 +/- 1.40 platelets (p less than 0.001 vs other studied groups). Platelet aggregate size was found not to be related to sex, age, medication, or coronary risk factors. Unstable angina may thus be a unique entity in ischemic heart disease concerning its platelet behavior, demonstrated in this study by the increased size of peripheral platelet aggregates, which may have pathogenetic, diagnostic, and eventual therapeutic implications.

pubmed23n0125_13745

Simultaneous occurrence of type A and type B Wolff-Parkinson-White conduction abnormalities: case report.

A 50 year-old woman with recurrent palpitations presented with electrocardiographic changes of simultaneous type A and type B Wolff-Parkinson-White conduction abnormalities suggesting the presence of multiple anomalous pathways.

Simultaneous occurrence of type A and type B Wolff-Parkinson-White conduction abnormalities: case report. A 50 year-old woman with recurrent palpitations presented with electrocardiographic changes of simultaneous type A and type B Wolff-Parkinson-White conduction abnormalities suggesting the presence of multiple anomalous pathways.

pubmed23n0125_13746

Replacement of ruptured aneurysm of arteriovenous fistula by segmental frozen venous allograft: a case report.

A ruptured aneurysm of an arteriovenous fistula created for chronic hemodialysis was replaced by a segment of frozen saphenous vein allograft. Fifteen months following the procedure the graft is patent. Frozen vein allografts should be considered as an appropriate option for the reconstruction of disrupted A-V fistulae.

Replacement of ruptured aneurysm of arteriovenous fistula by segmental frozen venous allograft: a case report. A ruptured aneurysm of an arteriovenous fistula created for chronic hemodialysis was replaced by a segment of frozen saphenous vein allograft. Fifteen months following the procedure the graft is patent. Frozen vein allografts should be considered as an appropriate option for the reconstruction of disrupted A-V fistulae.

pubmed23n0125_13747

Electrophoretic polymorphism and molecular structure of equine C3.

Plasma or serum samples from 12 Arabian and 181 standardbred horses have been typed using an immunofixation technique to determine electrophoretic polymorphism of equine third complement component (C3). Six distinctly different electrophoretic patterns of equine C3 have been recognized thus far. SDS PAGE analysis of equine C3/anti C3 complexes revealed that the submolecular structure comprised an alpha chain and beta chain of molecular weights approximately 118,000 and 63,000 daltons respectively. The molecular weights of the alpha and beta chains were similar in all electrophoretic variants tested. Family data derived from 73 mares, 21 stallions and 99 offspring suggested that the six electrophoretic phenotypes were inherited by means of three codominant alleles named C3-1, C3-2 and C3-3 at a single autosomal locus.

Electrophoretic polymorphism and molecular structure of equine C3. Plasma or serum samples from 12 Arabian and 181 standardbred horses have been typed using an immunofixation technique to determine electrophoretic polymorphism of equine third complement component (C3). Six distinctly different electrophoretic patterns of equine C3 have been recognized thus far. SDS PAGE analysis of equine C3/anti C3 complexes revealed that the submolecular structure comprised an alpha chain and beta chain of molecular weights approximately 118,000 and 63,000 daltons respectively. The molecular weights of the alpha and beta chains were similar in all electrophoretic variants tested. Family data derived from 73 mares, 21 stallions and 99 offspring suggested that the six electrophoretic phenotypes were inherited by means of three codominant alleles named C3-1, C3-2 and C3-3 at a single autosomal locus.

pubmed23n0125_13748

Genetic linkage between loci for a red cell alloantigen (U) and serum protease inhibitor (Pi) in the horse.

Preliminary evidence for the fifth autosomal linkage group in the horse, comprised of the loci for a red cell alloantigen (U) and serum protease inhibitor (Pi), was demonstrated by means of paternal half-sib groups in thoroughbred, standardbred and Arabian breeds. Recombination frequency in males was estimated to be 0.125 +/- 0.019.

Genetic linkage between loci for a red cell alloantigen (U) and serum protease inhibitor (Pi) in the horse. Preliminary evidence for the fifth autosomal linkage group in the horse, comprised of the loci for a red cell alloantigen (U) and serum protease inhibitor (Pi), was demonstrated by means of paternal half-sib groups in thoroughbred, standardbred and Arabian breeds. Recombination frequency in males was estimated to be 0.125 +/- 0.019.

pubmed23n0125_13749

Biochemical genetic variants in mice selectively bred for sensitivity or resistance to ethanol-induced sedation.

The distribution of biochemical genetic variants was examined among eight inbred strains of mice, which served as contributors to a heterogeneous stock of mice (HS), and in short-sleep (SS) and long-sleep (LS) mice, selectively bred from the HS stock for differential ethanol sensitivity. Fifteen loci for enzymes of alcohol and aldehyde metabolism, as well as 12 other biochemical loci, were investigated. Thirteen of these loci exhibited allelic variation between strains, of which six were separately fixed in the SS and LS mice. Comparisons of genetic similarity coefficients, based upon the distributions of allelic variants for the loci examined, with behavioural sensitivities (sleep-time) to an acute dose of ethanol for the inbred and selected strains of mice, indicated no correlations between these data. This suggests that this collective group of loci are not useful indicators of the genes selectively bred in the SS and LS strains, which are responsible for the differential sensitivities to acute doses of ethanol.

Biochemical genetic variants in mice selectively bred for sensitivity or resistance to ethanol-induced sedation. The distribution of biochemical genetic variants was examined among eight inbred strains of mice, which served as contributors to a heterogeneous stock of mice (HS), and in short-sleep (SS) and long-sleep (LS) mice, selectively bred from the HS stock for differential ethanol sensitivity. Fifteen loci for enzymes of alcohol and aldehyde metabolism, as well as 12 other biochemical loci, were investigated. Thirteen of these loci exhibited allelic variation between strains, of which six were separately fixed in the SS and LS mice. Comparisons of genetic similarity coefficients, based upon the distributions of allelic variants for the loci examined, with behavioural sensitivities (sleep-time) to an acute dose of ethanol for the inbred and selected strains of mice, indicated no correlations between these data. This suggests that this collective group of loci are not useful indicators of the genes selectively bred in the SS and LS strains, which are responsible for the differential sensitivities to acute doses of ethanol.

pubmed23n0125_13750

Genetic variations in blood proteins within and between Spanish dairy sheep breeds.

Eleven flocks of three Spanish sheep breeds have been studied: four Churra, four Lacha and three Manchega. Genetic variations between flocks of the same and different breeds have been analysed, using eight genetic blood systems and a wide range of statistical methods. The results show that the Churra, Lacha and Manchega breeds present significant differences between each other. Significant intraracial variations have been observed in Churras and Lachas, although they were almost non-existent in Manchegas. In some cases the genetic distances between flocks of the same breed were greater than the distances between flocks of different breeds. The data indicate that the closest breeds genetically are Lacha and Manchega.

Genetic variations in blood proteins within and between Spanish dairy sheep breeds. Eleven flocks of three Spanish sheep breeds have been studied: four Churra, four Lacha and three Manchega. Genetic variations between flocks of the same and different breeds have been analysed, using eight genetic blood systems and a wide range of statistical methods. The results show that the Churra, Lacha and Manchega breeds present significant differences between each other. Significant intraracial variations have been observed in Churras and Lachas, although they were almost non-existent in Manchegas. In some cases the genetic distances between flocks of the same breed were greater than the distances between flocks of different breeds. The data indicate that the closest breeds genetically are Lacha and Manchega.

pubmed23n0125_13751

Effect of swine lymphocyte antigen haplotypes on birth and weaning weights in pigs.

Birth weights of 708 live piglets and weaning weights of 566 piglets were used to investigate the effect of the swine lymphocyte antigen (SLA) complex on these traits in Large White pigs. Piglets were from litters of a long-term selection experiment to measure response for selection to increase litter size. SLA haplotypes were determined using conventional class I antisera. A total of 14 haplotypes were detected. The effect of SLA haplotype on birth and weaning weights was investigated using a statistical model that included the effects of experimental group, sire, dam, sex and SLA haplotype. Results indicated that SLA class I haplotype 13.1.3 increased birth weights (P less than 0.10) and significantly increased weaning weights (P less than 0.01). This effect of haplotype 13.1.3 on weaning weight was 605 +/- 215 g (0.3 standard deviations). SLA class I homozygosity did not appear to affect birth and weaning weights. These results suggest that the SLA complex plays an important role in early growth in the pig and that further study of SLA effects on growth and reproduction are warranted.

Effect of swine lymphocyte antigen haplotypes on birth and weaning weights in pigs. Birth weights of 708 live piglets and weaning weights of 566 piglets were used to investigate the effect of the swine lymphocyte antigen (SLA) complex on these traits in Large White pigs. Piglets were from litters of a long-term selection experiment to measure response for selection to increase litter size. SLA haplotypes were determined using conventional class I antisera. A total of 14 haplotypes were detected. The effect of SLA haplotype on birth and weaning weights was investigated using a statistical model that included the effects of experimental group, sire, dam, sex and SLA haplotype. Results indicated that SLA class I haplotype 13.1.3 increased birth weights (P less than 0.10) and significantly increased weaning weights (P less than 0.01). This effect of haplotype 13.1.3 on weaning weight was 605 +/- 215 g (0.3 standard deviations). SLA class I homozygosity did not appear to affect birth and weaning weights. These results suggest that the SLA complex plays an important role in early growth in the pig and that further study of SLA effects on growth and reproduction are warranted.

pubmed23n0125_13752

Rabbit plasma pretransferrin system: evidence for three new alleles.

Three additional pretransferrin types have been identified by one-dimensional PAGE technique in New Zealand White and Californian rabbits. The six alleles are designated PrtA, PrtB, PrtC, PrtD, PrtE and PrtF. It is likely that three of these six alleles are identical to those reported previously.

Rabbit plasma pretransferrin system: evidence for three new alleles. Three additional pretransferrin types have been identified by one-dimensional PAGE technique in New Zealand White and Californian rabbits. The six alleles are designated PrtA, PrtB, PrtC, PrtD, PrtE and PrtF. It is likely that three of these six alleles are identical to those reported previously.

pubmed23n0125_13753

Linkage analysis of loci controlling blood groups and the rectovaginal constriction syndrome in Jersey cattle.

The possibility of linkage between the recessive gene controlling the rectovaginal constriction (RVC) syndrome in Jersey cattle and 13 loci controlling blood groups and polymorphic proteins was studied. No evidence of close to moderate linkage was found between the RVC locus and any of the systems A, B, C, F, L, S, Z, R', Hb, Tf, Am-1 and Ca. No definite conclusion was possible with the M system.

Linkage analysis of loci controlling blood groups and the rectovaginal constriction syndrome in Jersey cattle. The possibility of linkage between the recessive gene controlling the rectovaginal constriction (RVC) syndrome in Jersey cattle and 13 loci controlling blood groups and polymorphic proteins was studied. No evidence of close to moderate linkage was found between the RVC locus and any of the systems A, B, C, F, L, S, Z, R', Hb, Tf, Am-1 and Ca. No definite conclusion was possible with the M system.

pubmed23n0125_13754

A standard method of intermittent inhaled therapy via a jet nebulizer.

Current methods of inhaled therapy using gas-propelled nebulizers lack standardization. The frequent use of nebulized therapy in a continuous flow may be the major reason for the inconsistency of the actual fraction of nebulized drug delivered to the patient. In an effort to achieve consistency in the amount of drug delivered to the patient, we evaluated an intermittent system of nebulized therapy, using a fingertip controlled nebulizer, in 18 adolescent asthmatics. This mode of delivery was found to be highly efficient, providing a nebulized fraction of more than 70% of the initial volumes used (0.75 to 2.6 mL). Initial volumes of 0.9 mL and above were equally efficacious, indicating no further benefit of using higher volumes. Nebulized fractions were found to be highly consistent from one patient to another (+/- 1 SD of 14.1%). We conclude that control of the actual dose delivered to the patient can be best achieved through an intermittent mode of delivery.

A standard method of intermittent inhaled therapy via a jet nebulizer. Current methods of inhaled therapy using gas-propelled nebulizers lack standardization. The frequent use of nebulized therapy in a continuous flow may be the major reason for the inconsistency of the actual fraction of nebulized drug delivered to the patient. In an effort to achieve consistency in the amount of drug delivered to the patient, we evaluated an intermittent system of nebulized therapy, using a fingertip controlled nebulizer, in 18 adolescent asthmatics. This mode of delivery was found to be highly efficient, providing a nebulized fraction of more than 70% of the initial volumes used (0.75 to 2.6 mL). Initial volumes of 0.9 mL and above were equally efficacious, indicating no further benefit of using higher volumes. Nebulized fractions were found to be highly consistent from one patient to another (+/- 1 SD of 14.1%). We conclude that control of the actual dose delivered to the patient can be best achieved through an intermittent mode of delivery.

pubmed23n0125_13755

Increased serum IgE antibodies in institutionalized asthmatic children after a transient return home. The role of house dust mite allergens in the home as a trigger of asthmatic attacks in mite-sensitive patients.

Increased serum IgE antibodies to Dermatophagoides pteronyssinus, a house dust mite, were observed in 12 of 13 institutionalized mite-sensitive asthmatic children after returning for four days to their own homes. These results implicate the house dust mite as a trigger of asthmatic attacks in these patients.

Increased serum IgE antibodies in institutionalized asthmatic children after a transient return home. The role of house dust mite allergens in the home as a trigger of asthmatic attacks in mite-sensitive patients. Increased serum IgE antibodies to Dermatophagoides pteronyssinus, a house dust mite, were observed in 12 of 13 institutionalized mite-sensitive asthmatic children after returning for four days to their own homes. These results implicate the house dust mite as a trigger of asthmatic attacks in these patients.

pubmed23n0125_13756

Serum IgE in newborns undergoing exchange transfusion.

Serum IgE was studied in a group of 20 newborns, ten males and ten females, with neonatal non-immunologic jaundice before and after exchange transfusion and in a control group without jaundice or other immunologic problems. Serum IgE was significantly higher in the study group than in the control group (P less than .005). After exchange transfusion there was a further increase (P less than .005). These changes do not seem to be due to the IgE content of the transfused blood. It is suggested that neonatal jaundice and/or exchange transfusion may stimulate the IgE synthesis in the newborn.

Serum IgE in newborns undergoing exchange transfusion. Serum IgE was studied in a group of 20 newborns, ten males and ten females, with neonatal non-immunologic jaundice before and after exchange transfusion and in a control group without jaundice or other immunologic problems. Serum IgE was significantly higher in the study group than in the control group (P less than .005). After exchange transfusion there was a further increase (P less than .005). These changes do not seem to be due to the IgE content of the transfused blood. It is suggested that neonatal jaundice and/or exchange transfusion may stimulate the IgE synthesis in the newborn.

pubmed23n0125_13757

A comparison of the immune response to immunotherapy with polymerized grass allergen and monomeric grass allergen.

This study compares the immune response of ten patients treated with polymerized grass (PG) immunotherapy with that of 12 patients treated with conventional monomer grass (MG) immunotherapy. The patients treated with PG immunotherapy received a mean cumulative dose of 44,840 protein nitrogen units (PNU), and those treated with MG immunotherapy received a mean cumulative dose of 46,083 PNU. Total antibody binding of perennial rye grass groups I, II, and III (RGGI, RGGII, and RGGIII) was measured in the serum of each individual. In addition, IgG titers to partially purified extracts of Bermuda, timothy, and orchard grass were also determined. A significant increase in the total antibody binding of RGGI, as well as an increase in the mean IgG titer to Bermuda, timothy, and orchard grass was demonstrated after treatment in the patients who had received PG immunotherapy. The mean total antibody binding of RGGII and RGGIII also increased in these patients, although not significantly. No significant difference in the mean total antibody binding of RGGI, RGGII, or RGGIII nor in the mean IgG titer to the three grass extracts was found in the sera of the two groups of patients. This study demonstrates a comparable immune response between immunotherapy with PG and MG, and retention of antigenic determinants during the polymerization process.

A comparison of the immune response to immunotherapy with polymerized grass allergen and monomeric grass allergen. This study compares the immune response of ten patients treated with polymerized grass (PG) immunotherapy with that of 12 patients treated with conventional monomer grass (MG) immunotherapy. The patients treated with PG immunotherapy received a mean cumulative dose of 44,840 protein nitrogen units (PNU), and those treated with MG immunotherapy received a mean cumulative dose of 46,083 PNU. Total antibody binding of perennial rye grass groups I, II, and III (RGGI, RGGII, and RGGIII) was measured in the serum of each individual. In addition, IgG titers to partially purified extracts of Bermuda, timothy, and orchard grass were also determined. A significant increase in the total antibody binding of RGGI, as well as an increase in the mean IgG titer to Bermuda, timothy, and orchard grass was demonstrated after treatment in the patients who had received PG immunotherapy. The mean total antibody binding of RGGII and RGGIII also increased in these patients, although not significantly. No significant difference in the mean total antibody binding of RGGI, RGGII, or RGGIII nor in the mean IgG titer to the three grass extracts was found in the sera of the two groups of patients. This study demonstrates a comparable immune response between immunotherapy with PG and MG, and retention of antigenic determinants during the polymerization process.

pubmed23n0125_13758

Mechanics of the otolith organ--dynamic response.

The otolith organs are the linear motion sensors of the mammalian system. As part of the vestibular system, these small organs are located in the inner ear. Mathematically modeled, they consist of an overdamped second-order system with elastic, viscous damping and mass elements. The governing equations of motion which describe the relative velocity of the mass with respect to the skull consist of a set of three coupled partial integral-differential equations. When these equations are nondimensionalized, they yield two nondimensional parameters which characterize the dynamic response of the system. These nondimensional equations are solved numerically for the relative displacement of the otolith mass for various values of the two nondimensional parameters. The solutions generated are for a step change in skull velocity. These solutions indicate that the end organ upper breakpoint frequency is at least one order of magnitude higher than previously measured experimental values determined by first-order neuron recordings.

Mechanics of the otolith organ--dynamic response. The otolith organs are the linear motion sensors of the mammalian system. As part of the vestibular system, these small organs are located in the inner ear. Mathematically modeled, they consist of an overdamped second-order system with elastic, viscous damping and mass elements. The governing equations of motion which describe the relative velocity of the mass with respect to the skull consist of a set of three coupled partial integral-differential equations. When these equations are nondimensionalized, they yield two nondimensional parameters which characterize the dynamic response of the system. These nondimensional equations are solved numerically for the relative displacement of the otolith mass for various values of the two nondimensional parameters. The solutions generated are for a step change in skull velocity. These solutions indicate that the end organ upper breakpoint frequency is at least one order of magnitude higher than previously measured experimental values determined by first-order neuron recordings.

pubmed23n0125_13759

A model of insulin delivery by a controlled release micropump.

A model has been developed to describe the delivery of insulin from a controlled release micropump (CRM). Basal delivery was provided by diffusion due to a concentration difference driving force across the CRM. This was modelled by considering the CRM to be a series of one-dimensional steady-state diffusion resistances. This delivery model was used to size prototypes and identify the piston, foam and the pump outlet as the controlling resistances to basal insulin transport. Augmented delivery by the CRM was achieved by repeated compression of a foam disk by a mild steel piston which was driven by a solenoid (tested voltage range 0-173 V DC; 5 msec "on" time; frequency 20-40 min-1). The increased delivery was attributed to the combination of mixing inside the pump barrel and displacement of barrel contents into the downstream reservoir. This action was approximated by a three-compartment model, which considered the CRM to consist of a well-mixed upstream reservoir and pump barrel (with a downstream reservoir) separated by two resistances: a constant upstream membrane resistance, (KmAm)-1, and a variable downstream mixing rate resistance, (Qd)-1. A least squares fit of the model to experimental data showed Qd to increase with the cube of the force on the piston and linearly with the compression frequency. In agreement with experimental results, the model predicted the upstream membrane to be rate controlling only at augmented pump resistances close to the value (KmAm)-1. These models were used to design an improved prototype (VIII) which is now being evaluated in vivo in pancreatectomized dogs for its efficacy in restoring and sustaining normoglycemia.

A model of insulin delivery by a controlled release micropump. A model has been developed to describe the delivery of insulin from a controlled release micropump (CRM). Basal delivery was provided by diffusion due to a concentration difference driving force across the CRM. This was modelled by considering the CRM to be a series of one-dimensional steady-state diffusion resistances. This delivery model was used to size prototypes and identify the piston, foam and the pump outlet as the controlling resistances to basal insulin transport. Augmented delivery by the CRM was achieved by repeated compression of a foam disk by a mild steel piston which was driven by a solenoid (tested voltage range 0-173 V DC; 5 msec "on" time; frequency 20-40 min-1). The increased delivery was attributed to the combination of mixing inside the pump barrel and displacement of barrel contents into the downstream reservoir. This action was approximated by a three-compartment model, which considered the CRM to consist of a well-mixed upstream reservoir and pump barrel (with a downstream reservoir) separated by two resistances: a constant upstream membrane resistance, (KmAm)-1, and a variable downstream mixing rate resistance, (Qd)-1. A least squares fit of the model to experimental data showed Qd to increase with the cube of the force on the piston and linearly with the compression frequency. In agreement with experimental results, the model predicted the upstream membrane to be rate controlling only at augmented pump resistances close to the value (KmAm)-1. These models were used to design an improved prototype (VIII) which is now being evaluated in vivo in pancreatectomized dogs for its efficacy in restoring and sustaining normoglycemia.

pubmed23n0125_13760

Fracture toughness of Kevlar 29/poly(methyl methacrylate) composite materials for surgical implantations.

A study of the fracture behaviour of Kevlar 29 reinforced dental cement is undertaken using both linear elastic and nonlinear elastic fracture mechanics techniques. Results from both approaches--of which the nonlinear elastic is believed to be more appropriate--indicate that a reinforcing effect is obtained for the fracture toughness even at very low fibre content. The flexural strength and modulus are apparently not improved, however, by the incorporation of Kevlar 29 fibres in the PMMA cement, probably because of the presence of voids, the poor fibre/matrix interfacial bonding and unsatisfying cement mixing practice. When compared to other PMMA composite cements, the present system appears to be probably more effective than carbon/PMMA, for example, in terms of fracture toughness. More experimental and analytical work is needed so as to optimize the mechanical properties with respect to structural parameters and cement preparation technique.

Fracture toughness of Kevlar 29/poly(methyl methacrylate) composite materials for surgical implantations. A study of the fracture behaviour of Kevlar 29 reinforced dental cement is undertaken using both linear elastic and nonlinear elastic fracture mechanics techniques. Results from both approaches--of which the nonlinear elastic is believed to be more appropriate--indicate that a reinforcing effect is obtained for the fracture toughness even at very low fibre content. The flexural strength and modulus are apparently not improved, however, by the incorporation of Kevlar 29 fibres in the PMMA cement, probably because of the presence of voids, the poor fibre/matrix interfacial bonding and unsatisfying cement mixing practice. When compared to other PMMA composite cements, the present system appears to be probably more effective than carbon/PMMA, for example, in terms of fracture toughness. More experimental and analytical work is needed so as to optimize the mechanical properties with respect to structural parameters and cement preparation technique.

pubmed23n0125_13761

Protein A in Staphylococcus intermedius isolates from dogs and cats.

The presence and quantity of extracellular and cell-bound protein A of Staphylococcus intermedius isolates from dogs and cats were determined, using an enzyme-linked immunoglobulin-binding assay. Horseradish peroxidase-conjugated rabbit anti-bovine immunoglobulin G purified by affinity chromatography was reacted with whole cell and supernatant fractions of S intermedius (n = 139), a protein A-producing strain of S aureus, and a protein A-deficient strain of S epidermidis. Extracellular protein A was found in 118 (84.9%) of 139 isolates of S intermedius. Most (69/118; 58.5%) of these isolates produced greater than 0.2 micrograms of extracellular protein A/ml. Cell-bound protein A was found in 6 (4.3%) of 139 isolates. Only 1 of these isolates contained cell-bound protein A exclusively. The other 5 isolates produced significantly greater amounts of extracellular protein A than cell-bound protein A. Additionally, greater than 96% of extracellular protein A could be removed from supernatants by adsorption with agarose gel containing immunoglobulin G.

Protein A in Staphylococcus intermedius isolates from dogs and cats. The presence and quantity of extracellular and cell-bound protein A of Staphylococcus intermedius isolates from dogs and cats were determined, using an enzyme-linked immunoglobulin-binding assay. Horseradish peroxidase-conjugated rabbit anti-bovine immunoglobulin G purified by affinity chromatography was reacted with whole cell and supernatant fractions of S intermedius (n = 139), a protein A-producing strain of S aureus, and a protein A-deficient strain of S epidermidis. Extracellular protein A was found in 118 (84.9%) of 139 isolates of S intermedius. Most (69/118; 58.5%) of these isolates produced greater than 0.2 micrograms of extracellular protein A/ml. Cell-bound protein A was found in 6 (4.3%) of 139 isolates. Only 1 of these isolates contained cell-bound protein A exclusively. The other 5 isolates produced significantly greater amounts of extracellular protein A than cell-bound protein A. Additionally, greater than 96% of extracellular protein A could be removed from supernatants by adsorption with agarose gel containing immunoglobulin G.

pubmed23n0125_13762

Immunoreactive protein antigens of Mycoplasma pulmonis in experimentally infected rats.

Mycoplasma pulmonis was cultured in modified Hayflick's medium, washed in 0.25 M NaCl, and solubilized by 2.5% sodium dodecyl sulfate. Protein antigens of M pulmonis separated by polyacrylamide-gel electrophoresis were blotted onto nitrocellulose strips. Specific-pathogen-free rats were inoculated intranasally with M pulmonis. The serum samples of these rats were obtained periodically and used to react with fractionated M pulmonis antigens which were fixed on the nitrocellulose strips. The antigen-antibody reactions were further recognized by 125I-labeled antiglobulin. Detection of immunoreactive antigens was obtained by autoradiography. Antibody response was not detected in serum obtained 7 days after rats were inoculated, and by 14 days, a slight response to several proteins was found. At 28 days after rats were inoculated, many immunoreactive antigens were detected. Generally, antibodies against antigens of moderate to low molecular weight appeared early in the infection, and antibodies against antigens of high molecular weight appeared late. Important immunoreactive antigens thus identified can readily be distinguished from more than 58 different M pulmonis antigens detectable by sodium dodecyl sulfate polyacrylamide-gel electrophoresis. The humoral antibody response was measured by an enzyme-linked immunosorbent assay. The immunoglobulin G antibodies were initially detected at low level at 7 days after rats were inoculated. These humoral antibody responses reached maximum by 28 days. The increase in serum antibody titer corresponded with numbers of immunoreactive antigens detected by immunoradio-binding assay. The information gained by this investigation may improve our understanding of the antigenicity of M pulmonis and the immune response of rats exposed to M pulmonis.

Immunoreactive protein antigens of Mycoplasma pulmonis in experimentally infected rats. Mycoplasma pulmonis was cultured in modified Hayflick's medium, washed in 0.25 M NaCl, and solubilized by 2.5% sodium dodecyl sulfate. Protein antigens of M pulmonis separated by polyacrylamide-gel electrophoresis were blotted onto nitrocellulose strips. Specific-pathogen-free rats were inoculated intranasally with M pulmonis. The serum samples of these rats were obtained periodically and used to react with fractionated M pulmonis antigens which were fixed on the nitrocellulose strips. The antigen-antibody reactions were further recognized by 125I-labeled antiglobulin. Detection of immunoreactive antigens was obtained by autoradiography. Antibody response was not detected in serum obtained 7 days after rats were inoculated, and by 14 days, a slight response to several proteins was found. At 28 days after rats were inoculated, many immunoreactive antigens were detected. Generally, antibodies against antigens of moderate to low molecular weight appeared early in the infection, and antibodies against antigens of high molecular weight appeared late. Important immunoreactive antigens thus identified can readily be distinguished from more than 58 different M pulmonis antigens detectable by sodium dodecyl sulfate polyacrylamide-gel electrophoresis. The humoral antibody response was measured by an enzyme-linked immunosorbent assay. The immunoglobulin G antibodies were initially detected at low level at 7 days after rats were inoculated. These humoral antibody responses reached maximum by 28 days. The increase in serum antibody titer corresponded with numbers of immunoreactive antigens detected by immunoradio-binding assay. The information gained by this investigation may improve our understanding of the antigenicity of M pulmonis and the immune response of rats exposed to M pulmonis.

pubmed23n0125_13763

Genotypic and phenotypic variation of biotypes coexisting in the Hickman strain of Newcastle disease virus.

Many Newcastle disease virus strains are composed of several biotypes which coexist in the wild and in laboratory cultures. We have studied some of the phenotypic and genotypic properties of 6 virus clones from the coexisting biotypes of the Hickman strain of Newcastle disease virus. These clones were readily distinguishable from the parent virus strain and from each other by their RNA fingerprints. Fingerprints of the most virulent clones (Hi/LC, Hi/MC, and Hi/LR) contained 61% to 77% of the oligoribonucleotides present in the fingerprint of the Hickman parent strain. None of the clones killed chickens as rapidly as did the parent strain, although some clones killed embryonating eggs as rapidly as did the parent strain.

Genotypic and phenotypic variation of biotypes coexisting in the Hickman strain of Newcastle disease virus. Many Newcastle disease virus strains are composed of several biotypes which coexist in the wild and in laboratory cultures. We have studied some of the phenotypic and genotypic properties of 6 virus clones from the coexisting biotypes of the Hickman strain of Newcastle disease virus. These clones were readily distinguishable from the parent virus strain and from each other by their RNA fingerprints. Fingerprints of the most virulent clones (Hi/LC, Hi/MC, and Hi/LR) contained 61% to 77% of the oligoribonucleotides present in the fingerprint of the Hickman parent strain. None of the clones killed chickens as rapidly as did the parent strain, although some clones killed embryonating eggs as rapidly as did the parent strain.

pubmed23n0125_13764

Cytotoxin (leukotoxin) production by Pasteurella haemolytica: requirement for an iron-containing compound.

In studies of Pasteurella haemolytica type 1 cytotoxin, filter-sterilized culture supernatants from organisms grown in RPMI-1640 tissue culture medium generally have been used. Supplementation of the medium with 7% bovine fetal serum was shown to be necessary for maximal cytotoxin production, as measured by percentage of bovine peripheral blood leukocytes that were killed. The serum-induced increase in cytotoxicity could not be explained simply by a greater percentage of increase in the number of viable organisms produced in the enriched medium. There also was no correlation between encapsulation of the organisms and cytotoxin production. Several natural iron-containing proteins including transferrin, lactoferrin, conalbumin, and hemoglobin stimulated cytotoxin production in lieu of bovine fetal serum, leading to the conclusion that one function of serum supplementation may be to increase the medium's iron concentration. A number of additional iron-containing and iron-chelating compounds were tested, with the conclusion that the iron concentration of the growth medium, as well as the presence of a suitable carrier molecule, may be critical for efficient cytotoxin production by P haemolytica.

Cytotoxin (leukotoxin) production by Pasteurella haemolytica: requirement for an iron-containing compound. In studies of Pasteurella haemolytica type 1 cytotoxin, filter-sterilized culture supernatants from organisms grown in RPMI-1640 tissue culture medium generally have been used. Supplementation of the medium with 7% bovine fetal serum was shown to be necessary for maximal cytotoxin production, as measured by percentage of bovine peripheral blood leukocytes that were killed. The serum-induced increase in cytotoxicity could not be explained simply by a greater percentage of increase in the number of viable organisms produced in the enriched medium. There also was no correlation between encapsulation of the organisms and cytotoxin production. Several natural iron-containing proteins including transferrin, lactoferrin, conalbumin, and hemoglobin stimulated cytotoxin production in lieu of bovine fetal serum, leading to the conclusion that one function of serum supplementation may be to increase the medium's iron concentration. A number of additional iron-containing and iron-chelating compounds were tested, with the conclusion that the iron concentration of the growth medium, as well as the presence of a suitable carrier molecule, may be critical for efficient cytotoxin production by P haemolytica.

pubmed23n0125_13765

Resistance of preimplantation bovine embryos to infection with Brucella abortus.

Preimplantation bovine embryos were exposed in vitro to Brucella abortus to determine if the bacteria would adhere to zona pellucida (ZP)-intact embryos or adhere to or infect ZP-free embryos. Brucella abortus was not isolated from ZP-intact or ZP-free groups of embryos after 10 sequential antibiotic-free washings. Brucella abortus was isolated from all groups containing ZP-defective embryos after the exposure period and washing. Detrimental effects on healthy in vitro development of embryos were not observed.

Resistance of preimplantation bovine embryos to infection with Brucella abortus. Preimplantation bovine embryos were exposed in vitro to Brucella abortus to determine if the bacteria would adhere to zona pellucida (ZP)-intact embryos or adhere to or infect ZP-free embryos. Brucella abortus was not isolated from ZP-intact or ZP-free groups of embryos after 10 sequential antibiotic-free washings. Brucella abortus was isolated from all groups containing ZP-defective embryos after the exposure period and washing. Detrimental effects on healthy in vitro development of embryos were not observed.

pubmed23n0125_13766

Prevalence of Histophilus ovis and Actinobacillus seminis in the genital tract of sheep.

Histophilus ovis and Actinobacillus seminis were isolated from the preputial cavity of 6-month-old rams and the vagina of 6-month-old ewes at a substantially higher rate than that in mature (greater than 2 years old) rams and ewes. These organisms appeared to be a transitory component of the ovine genital flora, the prevalence of which was associated with age regardless of gender. Additional evaluation of the recoverability of H ovis and A seminis from the preputial cavity of rams from birth to 1 year of age indicated that the isolation rate from rams and predominance of the organisms in the preputial cavity differed greatly over this age period. These organisms were not recoverable until ram lambs were 12 weeks of age and were most prevalent at 20 weeks of age, after which recoverability of H ovis and A seminis from the preputial cavity steadily decreased, continuing through the time of the last evaluation at 1 year of age. The time period with which these organisms can be isolated from the preputial cavity is closely correlated with the time period when epididymitis associated with these organisms develops, and may be an important factor in the pathogenesis of epididymitis.

Prevalence of Histophilus ovis and Actinobacillus seminis in the genital tract of sheep. Histophilus ovis and Actinobacillus seminis were isolated from the preputial cavity of 6-month-old rams and the vagina of 6-month-old ewes at a substantially higher rate than that in mature (greater than 2 years old) rams and ewes. These organisms appeared to be a transitory component of the ovine genital flora, the prevalence of which was associated with age regardless of gender. Additional evaluation of the recoverability of H ovis and A seminis from the preputial cavity of rams from birth to 1 year of age indicated that the isolation rate from rams and predominance of the organisms in the preputial cavity differed greatly over this age period. These organisms were not recoverable until ram lambs were 12 weeks of age and were most prevalent at 20 weeks of age, after which recoverability of H ovis and A seminis from the preputial cavity steadily decreased, continuing through the time of the last evaluation at 1 year of age. The time period with which these organisms can be isolated from the preputial cavity is closely correlated with the time period when epididymitis associated with these organisms develops, and may be an important factor in the pathogenesis of epididymitis.

pubmed23n0125_13767

Comparison of the antigens associated with saline solution, potassium thiocyanate, and sodium salicylate extracts of Pasteurella haemolytica serotype 1.

Pasteurella haemolytica antigenic extracts were made, using saline solution, potassium thiocyanate (KSCN), and sodium salicylate (SS) extraction procedures. Of the 3 techniques, saline solution extraction resulted in the lowest protein concentration and lowest ribonucleic acid-to-protein ratio. The extracts varied in protein:carbohydrate ratios, with the KSCN extract being highest and the saline solution extract the lowest. Each extract contained lipopolysaccharide, as determined by detectable quantities of 2-keto, 3-deoxyoctonate. The saline solution extract contained the fewest protein bands by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis, but contained the highest molecular weight proteins. All 3 extracts were reasonably similar antigenically, as detected by immunoblotting. Many of the protein bands present in the KSCN or SS extracts did not seem to be antigenic. Each extract was subjected to chromatofocusing, and the greatest antigenic peak, for each extract, failed to bind to the exchanger. These highly antigenic peaks, designated as saline solution, KSCN, or SS antigens, were similarly high in carbohydrate content, had similar antigenic-profiles, and contained high molecular weight (greater than 200,000) antigenic material, most likely carbohydrate in nature, as detected by immunoblotting. Inoculation of mice with 1 of the 3 extracts or the saline solution antigen resulted in marked antibody responses; however, protection against intraperitoneal challenge exposure to P haemolytica was minimal.

Comparison of the antigens associated with saline solution, potassium thiocyanate, and sodium salicylate extracts of Pasteurella haemolytica serotype 1. Pasteurella haemolytica antigenic extracts were made, using saline solution, potassium thiocyanate (KSCN), and sodium salicylate (SS) extraction procedures. Of the 3 techniques, saline solution extraction resulted in the lowest protein concentration and lowest ribonucleic acid-to-protein ratio. The extracts varied in protein:carbohydrate ratios, with the KSCN extract being highest and the saline solution extract the lowest. Each extract contained lipopolysaccharide, as determined by detectable quantities of 2-keto, 3-deoxyoctonate. The saline solution extract contained the fewest protein bands by sodium dodecyl sulfate-polyacrylamide-gel electrophoresis, but contained the highest molecular weight proteins. All 3 extracts were reasonably similar antigenically, as detected by immunoblotting. Many of the protein bands present in the KSCN or SS extracts did not seem to be antigenic. Each extract was subjected to chromatofocusing, and the greatest antigenic peak, for each extract, failed to bind to the exchanger. These highly antigenic peaks, designated as saline solution, KSCN, or SS antigens, were similarly high in carbohydrate content, had similar antigenic-profiles, and contained high molecular weight (greater than 200,000) antigenic material, most likely carbohydrate in nature, as detected by immunoblotting. Inoculation of mice with 1 of the 3 extracts or the saline solution antigen resulted in marked antibody responses; however, protection against intraperitoneal challenge exposure to P haemolytica was minimal.

pubmed23n0125_13768

Immunohistochemical staining patterns of canine eyes affected with chronic superficial keratitis.

Fourteen limbal biopsy specimens from 11 dogs with chronic superficial keratitis (CSK) were examined histologically and immunohistochemically. Ten of the 14 specimens had corneal epithelial hyperplasia and/or atrophy. Eleven of the 14 specimens had thickened epithelial basement membranes. Each specimen had cellular infiltration and lamellar disruption of the stroma. An avidin-biotin immunoperoxidase complex stain was used to detect immunoglobulin (Ig) deposition. Twelve of the 14 specimens stained positive for Ig. The staining pattern was consistent and characterized by diffuse deposition of stain in the superficial conjunctival stroma near the limbus. Four of the 12 Ig-positive specimens also stained positive in the superficial corneal stroma with 1 of these 4 also staining positive along the epithelial cell basement membrane. The diffuse pattern of stain deposition and the absence of staining of specific epithelial structures indicated that CSK is not a classical autoimmune disease similar to any disease in the pemphigus group or similar to systemic lupus erythematosus. Although the results may implicate CSK as an immune-mediated disease, nonspecific factors could not be ruled out.

Immunohistochemical staining patterns of canine eyes affected with chronic superficial keratitis. Fourteen limbal biopsy specimens from 11 dogs with chronic superficial keratitis (CSK) were examined histologically and immunohistochemically. Ten of the 14 specimens had corneal epithelial hyperplasia and/or atrophy. Eleven of the 14 specimens had thickened epithelial basement membranes. Each specimen had cellular infiltration and lamellar disruption of the stroma. An avidin-biotin immunoperoxidase complex stain was used to detect immunoglobulin (Ig) deposition. Twelve of the 14 specimens stained positive for Ig. The staining pattern was consistent and characterized by diffuse deposition of stain in the superficial conjunctival stroma near the limbus. Four of the 12 Ig-positive specimens also stained positive in the superficial corneal stroma with 1 of these 4 also staining positive along the epithelial cell basement membrane. The diffuse pattern of stain deposition and the absence of staining of specific epithelial structures indicated that CSK is not a classical autoimmune disease similar to any disease in the pemphigus group or similar to systemic lupus erythematosus. Although the results may implicate CSK as an immune-mediated disease, nonspecific factors could not be ruled out.

pubmed23n0125_13769

Immunotoxic effects of T-2 toxin on cell-mediated immunity to listeriosis in mice: comparison with cyclophosphamide.

Immunotoxic effects of T-2 toxin and cyclophosphamide on cell-mediated resistance were evaluated in mice exposed to Listeria monocytogenes infection. Mice were inoculated intraperitoneally with 4.0 X 10(5) (LD50) or 4.0 X 10(4) (nonlethal) L monocytogenes and were treated with 4.0 mg of T-2 toxin/kg of body weight or 180 mg of cyclophosphamide/kg. The immunosuppressive effect of the toxin and cyclophosphamide was indicated by the rapid growth of Listeria and significant (P less than 0.005) increases in mortality because of listeriosis. Necrosis and depletion of lymphoid tissue, lymphopenia, and significant (P less than 0.005) decreases in the influx and number of lymphocytes and macrophages occurred in Listeria-elicited peritoneal exudates and at sites of infection in the liver and spleen of the toxin- and cyclophosphamide-treated mice. Immunotoxic effects of T-2 toxin and cyclophosphamide were comparable and attributed primarily to the depletion of T lymphocytes and the subsequent failure of surviving immunologically committed T cells and T-cell dependent immune-activated macrophages to clear the host of bacteria.

Immunotoxic effects of T-2 toxin on cell-mediated immunity to listeriosis in mice: comparison with cyclophosphamide. Immunotoxic effects of T-2 toxin and cyclophosphamide on cell-mediated resistance were evaluated in mice exposed to Listeria monocytogenes infection. Mice were inoculated intraperitoneally with 4.0 X 10(5) (LD50) or 4.0 X 10(4) (nonlethal) L monocytogenes and were treated with 4.0 mg of T-2 toxin/kg of body weight or 180 mg of cyclophosphamide/kg. The immunosuppressive effect of the toxin and cyclophosphamide was indicated by the rapid growth of Listeria and significant (P less than 0.005) increases in mortality because of listeriosis. Necrosis and depletion of lymphoid tissue, lymphopenia, and significant (P less than 0.005) decreases in the influx and number of lymphocytes and macrophages occurred in Listeria-elicited peritoneal exudates and at sites of infection in the liver and spleen of the toxin- and cyclophosphamide-treated mice. Immunotoxic effects of T-2 toxin and cyclophosphamide were comparable and attributed primarily to the depletion of T lymphocytes and the subsequent failure of surviving immunologically committed T cells and T-cell dependent immune-activated macrophages to clear the host of bacteria.

pubmed23n0125_13770

Echocardiographic characterization of dilatation cardiomyopathy in the English cocker spaniel.

The echocardiographic characterization of a dilatation cardiomyopathy in small-breed dogs is reported. Twelve clinically healthy adult English Cocker Spaniel dogs (between 2 and 9 years old and weighing 11.5 to 15.4 kg [mean 12.9 +/- 1.00 kg]) from a kennel population with a history of cardiomyopathy were assessed, using M-mode echocardiography. The dogs were selected on ECG and/or radiographic evidence of ventricular enlargement. Nine dogs had R-wave amplitude in lead 11 of greater than 3.0 mV. Two dogs had an unusual right-axis deviation, the result of deep Q waves in the limb leads and deep S waves in chest leads CV6LL and CV6LU, indicating that there was right ventricular enlargement. All dogs had increased end-systolic dimensions (mean 3.0 +/- 0.6 cm). End-diastolic dimensions were increased in 9 dogs (mean 4.0 +/- 0.5 cm), and there was a decrease of left ventricular (LV) function as measured by fractional shortening in 8 dogs. Mean fractional shortening for the 12 dogs was 25.4 +/- 5.7%. There was significant correlation between LV dimensions and age at echocardiographic assessment, indicating that LV dilatation was progressive. Three of the oldest dogs had severe dilatation of the LV, and in 2 of these, LV function was severely decreased. Left ventricular function in the 3rd dog, however, was within the acceptable range. Fractional shortening and thickness of the LV caudal wall and interventricular septum were significantly correlated (P less than 0.01 for interventricular system and P less than 0.05 for LV caudal wall).(ABSTRACT TRUNCATED AT 250 WORDS)

Echocardiographic characterization of dilatation cardiomyopathy in the English cocker spaniel. The echocardiographic characterization of a dilatation cardiomyopathy in small-breed dogs is reported. Twelve clinically healthy adult English Cocker Spaniel dogs (between 2 and 9 years old and weighing 11.5 to 15.4 kg [mean 12.9 +/- 1.00 kg]) from a kennel population with a history of cardiomyopathy were assessed, using M-mode echocardiography. The dogs were selected on ECG and/or radiographic evidence of ventricular enlargement. Nine dogs had R-wave amplitude in lead 11 of greater than 3.0 mV. Two dogs had an unusual right-axis deviation, the result of deep Q waves in the limb leads and deep S waves in chest leads CV6LL and CV6LU, indicating that there was right ventricular enlargement. All dogs had increased end-systolic dimensions (mean 3.0 +/- 0.6 cm). End-diastolic dimensions were increased in 9 dogs (mean 4.0 +/- 0.5 cm), and there was a decrease of left ventricular (LV) function as measured by fractional shortening in 8 dogs. Mean fractional shortening for the 12 dogs was 25.4 +/- 5.7%. There was significant correlation between LV dimensions and age at echocardiographic assessment, indicating that LV dilatation was progressive. Three of the oldest dogs had severe dilatation of the LV, and in 2 of these, LV function was severely decreased. Left ventricular function in the 3rd dog, however, was within the acceptable range. Fractional shortening and thickness of the LV caudal wall and interventricular septum were significantly correlated (P less than 0.01 for interventricular system and P less than 0.05 for LV caudal wall).(ABSTRACT TRUNCATED AT 250 WORDS)

pubmed23n0125_13771

Evaluation of plasma and buffy coat ascorbic acid concentrations in dogs before and after a 24-hour fast.

Samples of blood were taken from 15 female and 15 male research laboratory Beagles before and after they were fasted for 24 hours. The mean buffy coat ascorbic acid concentration was significantly higher in dogs after they were fasted than that before they were fasted. In contrast, the mean plasma ascorbic acid concentration was significantly lower in dogs after they were fasted than that before they were fasted. The mean buffy coat ascorbic acid concentrations in blood samples of both fasted and nonfasted female Beagles was significantly greater than those of male Beagles, whereas the mean plasma concentrations of both fasted and nonfasted female Beagles was significantly lower than those of male Beagles. It was observed that whenever there was a decrease in plasma ascorbic acid concentration, there was an increase in buffy coat ascorbic acid concentration, regardless of fasting stress or sex difference.

Evaluation of plasma and buffy coat ascorbic acid concentrations in dogs before and after a 24-hour fast. Samples of blood were taken from 15 female and 15 male research laboratory Beagles before and after they were fasted for 24 hours. The mean buffy coat ascorbic acid concentration was significantly higher in dogs after they were fasted than that before they were fasted. In contrast, the mean plasma ascorbic acid concentration was significantly lower in dogs after they were fasted than that before they were fasted. The mean buffy coat ascorbic acid concentrations in blood samples of both fasted and nonfasted female Beagles was significantly greater than those of male Beagles, whereas the mean plasma concentrations of both fasted and nonfasted female Beagles was significantly lower than those of male Beagles. It was observed that whenever there was a decrease in plasma ascorbic acid concentration, there was an increase in buffy coat ascorbic acid concentration, regardless of fasting stress or sex difference.

pubmed23n0125_13772

Flow cytofluorometric characterization of bovine blood and milk leukocytes.

Flow cytometry and sorting proved to be a rapid method that facilitated the identification of different leukocyte populations in bovine blood and milk. After briefly incubating whole blood and milk samples in a hypotonic phosphate buffer, containing supravital acridine orange, 5 classes of leukocytes were found in the blood (lymphocytes, neutrophils, eosinophils, basophils, and monocytes) and 4 in the milk (lymphocytes, neutrophils, monocytes, and macrophages) by flow cytometry. Cells were morphologically identified by fluorescent microscopy after flow cytometric sorting and by light microscopy after Papanicolaous staining. Udder parenchymal and ductal tissue cells (secretory and epithelial cells) were not found in the milk samples evaluated. Large differences in the total and differential cell counts were found in the different milk secretions.

Flow cytofluorometric characterization of bovine blood and milk leukocytes. Flow cytometry and sorting proved to be a rapid method that facilitated the identification of different leukocyte populations in bovine blood and milk. After briefly incubating whole blood and milk samples in a hypotonic phosphate buffer, containing supravital acridine orange, 5 classes of leukocytes were found in the blood (lymphocytes, neutrophils, eosinophils, basophils, and monocytes) and 4 in the milk (lymphocytes, neutrophils, monocytes, and macrophages) by flow cytometry. Cells were morphologically identified by fluorescent microscopy after flow cytometric sorting and by light microscopy after Papanicolaous staining. Udder parenchymal and ductal tissue cells (secretory and epithelial cells) were not found in the milk samples evaluated. Large differences in the total and differential cell counts were found in the different milk secretions.

pubmed23n0125_13773

Breath hydrogen concentration and small intestinal malabsorption in calves.

Breath hydrogen concentrations were measured to assess intestinal carbohydrate malabsorption in preruminating calves. Oral administration of 1.25 g of lactulose (a nonabsorbable carbohydrate)/kg to calves produced breath hydrogen concentrations significantly (P less than 0.001) higher than values determined after calves were fed milk and before the treatment was given. This indicates that, in the calf, fermentation of nonabsorbed carbohydrates results in increased breath hydrogen values. To induce small intestinal malabsorption, chloramphenicol was administered orally at 50 mg/kg, 2 times a day, to 5 calves for 3 days. Before therapy was started, each calf was fitted with a duodenal cannula to facilitate collection of intestinal mucosal biopsy samples during treatment. Chloramphenicol therapy significantly (P less than 0.001) increased breath hydrogen concentrations from those values measured after calves were fed milk alone. Concurrently, chloramphenicol administration significantly decreased intestinal villous length (P less than 0.001) and D-xylose absorption (P less than 0.05), compared with those values before treatment was given. These results demonstrate that decreased intestinal absorptive capacity is associated with an increase in breath hydrogen concentrations and that breath hydrogen may be useful in evaluating malabsorption in calves with naturally occurring enteric disease.

Breath hydrogen concentration and small intestinal malabsorption in calves. Breath hydrogen concentrations were measured to assess intestinal carbohydrate malabsorption in preruminating calves. Oral administration of 1.25 g of lactulose (a nonabsorbable carbohydrate)/kg to calves produced breath hydrogen concentrations significantly (P less than 0.001) higher than values determined after calves were fed milk and before the treatment was given. This indicates that, in the calf, fermentation of nonabsorbed carbohydrates results in increased breath hydrogen values. To induce small intestinal malabsorption, chloramphenicol was administered orally at 50 mg/kg, 2 times a day, to 5 calves for 3 days. Before therapy was started, each calf was fitted with a duodenal cannula to facilitate collection of intestinal mucosal biopsy samples during treatment. Chloramphenicol therapy significantly (P less than 0.001) increased breath hydrogen concentrations from those values measured after calves were fed milk alone. Concurrently, chloramphenicol administration significantly decreased intestinal villous length (P less than 0.001) and D-xylose absorption (P less than 0.05), compared with those values before treatment was given. These results demonstrate that decreased intestinal absorptive capacity is associated with an increase in breath hydrogen concentrations and that breath hydrogen may be useful in evaluating malabsorption in calves with naturally occurring enteric disease.

pubmed23n0125_13774

Evaluation of lasalocid as a coccidiostat in calves: titration, efficacy, and comparison with monensin and decoquinate.

Two experiments were conducted to evaluate lasalocid as a coccidiostat in Holstein calves and to compare lasalocid with monensin and decoquinate. In experiment 1, calves in 3 groups (6 calves/group) were each inoculated with 500,000 sporulated oocysts, 88% of which were Eimeria bovis and 12% were E zuernii. Calves in each group were given lasalocid-medicated feed at 0.50 (group 3), 0.75 (group 4), or 1 mg/kg (group 5) of body weight/day for 45 days. Two control groups (6 calves/group) were also evaluated; calves in control group 2 were inoculated and nontreated, and calves in control group 1 were noninoculated and nontreated. At 0.50, 0.75, or 1 mg/kg/day, lasalocid was equally effective in preventing induced coccidiosis (E bovis and E zuernii) in calves. Compared with inoculated nontreated controls, treated calves had significantly (P less than 0.05) fewer oocysts in feces and had fewer clinical signs of coccidiosis from days 16 to 30 after inoculation. Experiment 2 was conducted to compare the effectiveness of monensin, lasalocid, and decoquinate for the prevention of experimentally induced coccidiosis. Calves (n = 48) were allotted into 4 groups (12 calves/group); each was inoculated orally with 275,000 sporulated oocysts, predominantly E bovis and E zuernii, and each was given nonmedicated feed (group 6) or feed medicated with 33 mg of lasalocid (group 7), decoquinate (group 8), or monensin (group 9)/kg of feed for 46 days. Calves given medicated rations had significantly (P less than 0.05) fewer oocysts in their feces and fewer clinical signs of coccidiosis than did calves given nonmedicated rations.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of lasalocid as a coccidiostat in calves: titration, efficacy, and comparison with monensin and decoquinate. Two experiments were conducted to evaluate lasalocid as a coccidiostat in Holstein calves and to compare lasalocid with monensin and decoquinate. In experiment 1, calves in 3 groups (6 calves/group) were each inoculated with 500,000 sporulated oocysts, 88% of which were Eimeria bovis and 12% were E zuernii. Calves in each group were given lasalocid-medicated feed at 0.50 (group 3), 0.75 (group 4), or 1 mg/kg (group 5) of body weight/day for 45 days. Two control groups (6 calves/group) were also evaluated; calves in control group 2 were inoculated and nontreated, and calves in control group 1 were noninoculated and nontreated. At 0.50, 0.75, or 1 mg/kg/day, lasalocid was equally effective in preventing induced coccidiosis (E bovis and E zuernii) in calves. Compared with inoculated nontreated controls, treated calves had significantly (P less than 0.05) fewer oocysts in feces and had fewer clinical signs of coccidiosis from days 16 to 30 after inoculation. Experiment 2 was conducted to compare the effectiveness of monensin, lasalocid, and decoquinate for the prevention of experimentally induced coccidiosis. Calves (n = 48) were allotted into 4 groups (12 calves/group); each was inoculated orally with 275,000 sporulated oocysts, predominantly E bovis and E zuernii, and each was given nonmedicated feed (group 6) or feed medicated with 33 mg of lasalocid (group 7), decoquinate (group 8), or monensin (group 9)/kg of feed for 46 days. Calves given medicated rations had significantly (P less than 0.05) fewer oocysts in their feces and fewer clinical signs of coccidiosis than did calves given nonmedicated rations.(ABSTRACT TRUNCATED AT 250 WORDS)

pubmed23n0125_13775

Anthelmintic treatment of pastured dairy cattle in California.

The relationship of anthelmintic treatment for subclinical gastrointestinal nematode parasitism with resultant milk production was studied on 3 California dairies where animals were maintained on a pasture-based nutritional program. All of the animals that started lactating (freshened) during a 12-month period participated in trials in which coumaphos was given as a feed top dressing to animals averaging 30 days into lactation. All of the animals that freshened during the subsequent 12-month period from 2 of the same 3 dairies participated in trials in which thiabendazole was administered at or within 2 weeks before freshening. Anthelmintic administration during 2 consecutive lactation cycles under the conditions of the present study resulted in no consistent reproducible significant changes in milk production.

Anthelmintic treatment of pastured dairy cattle in California. The relationship of anthelmintic treatment for subclinical gastrointestinal nematode parasitism with resultant milk production was studied on 3 California dairies where animals were maintained on a pasture-based nutritional program. All of the animals that started lactating (freshened) during a 12-month period participated in trials in which coumaphos was given as a feed top dressing to animals averaging 30 days into lactation. All of the animals that freshened during the subsequent 12-month period from 2 of the same 3 dairies participated in trials in which thiabendazole was administered at or within 2 weeks before freshening. Anthelmintic administration during 2 consecutive lactation cycles under the conditions of the present study resulted in no consistent reproducible significant changes in milk production.

pubmed23n0125_13776

Anthelmintic efficacy of febantel combined with praziquantel against Ancylostoma tubaeforme, Toxocara cati, and Taenia taeniaeformis in cats.

Forty cats, each harboring 2 or 3 parasitic infections (Ancylostoma tubaeforme, Toxocara cati, and/or Taenia taeniaeformis), were used to titrate the anthelmintic efficacy of a paste containing 3.4% febantel and 0.34% praziquantel. The cats were allotted into 4 groups (10 cats/group). For 3 consecutive days, the cats were given febantel/praziquantel at 5/0.5 mg/kg/day, 10/1 mg/kg/day, 15/1.5 mg/kg/day, or a blank paste vehicle (control) at 0.29 g/kg of body weight. The recommended dosage of 10 mg of febantel and 1 mg of praziquantel/kg cleared greater than or equal to 98% of the 3 helminth species.

Anthelmintic efficacy of febantel combined with praziquantel against Ancylostoma tubaeforme, Toxocara cati, and Taenia taeniaeformis in cats. Forty cats, each harboring 2 or 3 parasitic infections (Ancylostoma tubaeforme, Toxocara cati, and/or Taenia taeniaeformis), were used to titrate the anthelmintic efficacy of a paste containing 3.4% febantel and 0.34% praziquantel. The cats were allotted into 4 groups (10 cats/group). For 3 consecutive days, the cats were given febantel/praziquantel at 5/0.5 mg/kg/day, 10/1 mg/kg/day, 15/1.5 mg/kg/day, or a blank paste vehicle (control) at 0.29 g/kg of body weight. The recommended dosage of 10 mg of febantel and 1 mg of praziquantel/kg cleared greater than or equal to 98% of the 3 helminth species.