Datasets:

andersonbcdefg

/

pubmed_pairs_part2

like 0

Detection of herpes simplex virus infection of female genitalia by the peroxidase-antiperoxidase method alone or in conjunction with the Papanicolaou stain.

The sensitive peroxidase-antiperoxidase (PXAPX) method individually and in conjunction with the Papanicolaou (PAP) stain was used to detect herpes simplex virus (HSV) in specimens from human female genitalia. Initial studies using a model system of HSV-1 or HSV-2-infected Vero cells established (i) acetone as the most effective fixative, (ii) optimal dilutions of preimmunization and anti-HSV serum for differentiation of infected from noninfected cells, (iii) optimal concentration of 3,3'-diaminobenzidine tetrahydrochloride and H2O2 for maximal staining of infected cells with minimal background reaction, and (iv) removal of endogenous peroxidase with absolute MeOH. These various parameters, once established, were utilized in the PXAPX or PXAPX-PAP on human specimens from the vulva or cervix. In these specimens, examined by standard light microscopy, PXAPX-positive cells were dark brown with a single nucleus or multiple nuclei. By coupling the PAP to the PXAPX, detailed nuclear observations of PXAPX-positive cells were possible and revealed nuclear changes consistent with HSV infection, including syncytium formation, chromatin condensation, and an occasional Cowdry type A inclusion. The PXAPX and PXAPX-PAP correlated (r = 0.742) over a period of 72 h with HSV isolation from these samples.

The errors in assembly of MuLV in interferon treated cells.

Interferon treatment of JLSV-6 cells chronically infected with Rauscher MuLV leads to the formation of noninfectious particles (interferon virions) containing the structural proteins of env and gag genes as well as additional viral polypeptides. In the control virions the major glycoprotein detected is gp71, interferon virions contain in addition to gp71 and 85k dalton (gp85) glucosamine-containing, fucose-deficient glycoprotein which is recognized by antiserum to MuLV but not by the gp71 antiserum. The surface iodination of the intact virions indicates that both gp71 and gp85 are the major components of the external virions envelope. However, unlike the control virions in which gp71 associates with p15E (gp90), the gp71-p15E complex was not detected in interferon virions. The analysis of the iodinated proteins of the disrupted interferon virions revealed the presence of 85k and 65k dalton polypeptides preciptable with antiserum against MuLV, which are not present in the control virions. The difference in the polypeptide pattern of virions produced in the presence of interferon does not seem to be a consequence of the slowdown in the synthesis of viral proteins or their processing in the interferon-treated cells. Both the structural proteins of env and gag genes seem to be synthesized and processed at a comparable rate in the interferon-treated and -untreated cells. These results indicate an alteration of virus assembly in the presence of interferon.

Effects of propranolol and timolol on calcium uptake by sarcoplasmic reticulum vesicles.

The effects of the beta-adrenergic receptor blocking agents propranolol and timolol on the initial calcium uptake velocity of sarcoplasmic reticulum vesicles of rabbit skeletal muscle were studied. Racemic d- and l-propranolol had similar inhibitory effects on initial calcium uptake velocity, which was inhibited 50% by 5--7 X 10(-4) M racemic propranolol. Timolol was a much less potent inhibitor of initial calcim uptake velocity; 50% inhibition occurred at approximately 10(-2) M timolol. Both drugs inhibited maximal calcium uptake velocity; however, KCa (the Ca2+ concentration at which calcium uptake was half-maximal) was modified differently. Propranolol increased KCa, whereas timolol caused the KCa to decrease. Addition of either drug to an ongoing calcium uptake reaction at the time that calcium content became maximal caused renewed calcium uptake. The relative potencies of propranolol and timolol as negative inotropic agents are similar to their potencies as inhibitors of sarcoplasmic reticulum calcium uptake, but dissimilar to their beta-adrenergic receptor blocking potencies. Timolol, which has been reported to have less negative inotropic effect than propranolol, is approximately 5 time more potent than propranolol as a beta-adrenergic receptor blocking agent but 15 times less potent as an inhibitor of sarcoplasmic reticulum calcium uptake. Inhibition of sarcoplasmic reticulum calcium uptake may thus characterize negative inotropic potencies of new beta-adrenergic receptor blocking agents.

Chronotropic effects of atropine sulfate and methscopolamine bromide in normal subjects and patients undergoing cardiac catheterization.

The increase in heart rate caused by intravenous injections of atropine sulfate (AS) and methscopolamine bromide (MSB) were compared in 12 healthy volunteers. AS, 200 micrograms, or MSB, 100 micrograms, were injected every 3 min for five doses on successive days. Mean heart rate from the second and fifth injections of MSB was significantly greater than after corresponding injections of atropine. The ratio of equivalent doses of MSB and AS was found to be 1:3. Three subjects with AS and four subjects with MSB exhibited reduced heart rates after the first injection. Excellent correlation of plasma atropine concentrations and heart rate increase were obtained in two volunteers (correlation coefficient, r = 0.84). In the second study, AS, 180 micrograms, and MSB, 60 micrograms, were compared in 20 patients undergoing cardiac catheterization and coronary arteriography. Both drugs produced equivalent increments in heart rate. MSB produced a more predictable dose response than AS. Side effects were similar with both drugs when equivalent doses were used. This study confirms previous investigations that MSB can be titrated by intravenous injections to increase heart rate to desired levels. Absence of central nervous system stimulation by MSB suggests that it may be more appropriate for use in patients with myocardial infarction.

Isolation and characterization of free and membrane-bound polyribosomes from rabbit spinal cord.

A procedure is described for the preparation of free and membrane-bound polyribosomes from rabbit spinal cord. First, myelin and filaments are floated away from other subcellular components. The latter are then fractionated by differential centrifugation followed by sedimentation through a discontinuous sucrose gradient. The ribosomal fractions are characterized by their electron microscopic appearance, RNA/protein ratios and sedimentation profile in a linear sucrose gradient. Both membrane-bound and free polyribosomes are active in incorporating amino acids in a cell-free system, whether heterologous or homologous pH 5 enzyme fractions are employed. Autoradiographic analysis of translation products separated by electrophoresis on SDS-polyacrylamide gels shows that both ribosome fractions are active in the synthesis of a variety of proteins including polypeptides which comigrate with alpha- and beta-tubulins and actin. The utility of these polyribosome preparations for the cell-free study of protein biosynthesis is indicated by the high molecular weight of many of the translation products, and by the similarity of the electrophoretic pattern of translation products from the cell-free systems to the pattern of radioactive protein synthesized by rabbit spinal cord during the hour prior to sacrifice.

Neutral protease activity in lymphocytes of Lewis rats with acute experimental allergic encephalomyelitis.

Lymphocytes from popliteal and inguinal lymph nodes of Lewis rats with acute EAE as a result of injection of lyophilized guinea pig myelin in Freund's complete adjuvant exerted strong proteolytic activity at neutral pH toward myelin basic protein. After injection of myelin the level of proteolytic activity remained about the same as that in lymphocytes from Freund's adjuvant-injected controls until about day 10 after injection, just before the onset of paralytic symptoms; then the proteolytic activity increased to approximately double its former level. Myelin basic protein was hydrolyzed by whole lymphocytes, but more activity was unmasked by homogenization. Similar results were also obtained using lymphocytes from thymus of EAE and control animals. Lymphocytes with high levels of proteolytic activity were not absorbed by glass wool, did not stain with neutral red, nor did they phagocytose antibody-coated sheep red blood cells. Thymus and lymph node lymphocytes cleaved myelin basic protein to three major peptides and a fourth minor peptide, while spleen lymphocytes hydrolyzed basic protein at only one point resulting in two peptides whose molecular weights added up to that of myelin basic protein. The protease activity was inhibited by 5 X 10(-3) M p-chloromercuribenzoate and by phenylmethyl sulfonyl fluoride, TPCK, and soybean trypsin inhibitor, therefore the enzymatic activity probably depends on a serine residue and a sulfhydryl group. The bulk of the enzymatic activity is mostly membrane bound with the highest specific activity and total activity contained in a lysosomal-mitochondrial fraction. In view of the infiltration of lymphocytes into the brain substance in acute EAE, it is suggested that these cells may contribute to the destruction of myelin which is usually attributed to the monocyte or macrophage.

The development of an experimental model of subretinal neovascularization in disciform macular degeneration.

A reproducible model of subretinal neovascularization has been developed, an important first step in the study of disciform macular degeneration. The methodology and clinical description of subretinal neovascularization in the primate have been provided and histopathologic and electron microscopic correlations included. This model provides a most promising and useful mechanism for the evaluation of the many different factors and hypotheses that have been proposed for the genesis of the disciform response. In humans, it probably has more relevance to the traumatically induced disciform process, particularly that after laser therapy, than to the senile degenerative process. The purpose of this investigation is to develop the animal model by which many different theories regarding the pathogenesis of the disciform response can be tested. These initial studies relate to ischemia, hemorrhage, the inflammatory response, and position in relation to the center of the vascular-free zone as factors important in the development and evolution of subretinal neovascularization.

Hepatic microsomal induction and hepatic transport.

Hepatic microsomal induction (hexobarbital sleeping time, cytochrome P-450 and microsomal protein concentration, liver weight) and hepatic transport (hepatic uptake, biotransformation, biliary excretion) have been studied in rats. Phenobarbital pretreatment (75 mg/kg i.p. daily for 5 days) produced microsomal induction in the liver and enhanced biliary excretion of bromcresol green, eosine, bromsulphthaleine glutathione conjugate (BSP-GSH), amaranth and iodoxamic acid. However, biliary excretion of indocyanine green was unchanged after phenobarbital pretreatment. Biotransformation of bromsulphthalein (BSP) with glutathione was also increased by phenobarbital. The hepatic concentration of these organic anions was not influenced uniformly after phenobarbital pretreatment: the concentration of indocyanine green, bromcresol green, eosin and BSP-GSH in the liver was unchanged, that of amaranth and iodoxamic acid was enhanced following phenobarbital pretreatment. Investigation of the effect of pretreatment with other barbiturates showed that barbital, butobarbital, pentobarbital and amobarbital produced microsomal induction. Only baribtal and butobarbital stimulated biliary excretion of organic anions, whereas pentobarbital and amobarbital proved to be ineffective in this parameter. The results seem to indicate that the enhanced biliary excretion of exogenous organic anions produced by barbiturates is independent of microsomal enzyme induction.

The plasma kinetics of hydroxyethyl starch 350/0.60: a potential new adjunct for centrifugal leucapheresis.

A modified species of hydroxyethyl starch (HES 350/0.60) possessing a Mw- of 350,000 daltons combined with a molar hydroxyethyl group substitution (MS) of 0.60 (60 hydroxyethyl groups.100 glucose residues) was clinically assessed in seven normal subjects to determine the influence of these chemical modifications on intravascular clearance kinetics concomitantly with effects on the suspension stability (ESR) of blood. Following a standardised intravenous dose (30 gm.m2 BSA), the concentration of HES 350/0.60 in serum fell to half its peak value in 11.8 +/- 1.3 (SD) hours, while the ESR remained elevated for up to 12 hours post-injection. By adopting a Mw- of 350,000 daltons, the critical molecular weight (Cmw) of this colloid was surpassed, while the critical concentration (Cc), below which the suspension stability of blood is not affected, was shown to range between 0.3 and 0.5 gm.dl-1. In comparison to the present species of HES (Mw- 450,000 daltons, MS: 0.70) utilised as a sedimenting agent duirng centrifugal leucapheresis, HES 350,000/0.60 appears to affect the ESR in a similar manner, but is removed from the intravascular space approximately twice as rapidly. This more rapid clearance should be useful in avoiding cumulative build-up of HES in blood concomitant with reducing the total amount of intravascular H2O bound to this colloid, in normal and CML donors undergoing multiple cell collection procedures.

Thiamine intestinal transport and phosphorylation : a study in vitro of potential inhibitors of small intestinal thiamine-pyrophosphokinase using a crude enzymatic preparation.

Using as enzymatic source the cytoplasmatic fraction of enterocytes isolated from the rat small intestine, thiamine-pyrophosphokinase activity was studied with a radiometric method using [thiazole-2-(14)C] thiamine. The Km value for thiamine was 2.14 X 10(-6) M and V 0.87 nmol of thiamine pyrophosphate mg-1 protein h-1. Eleven thiamine structural analogs and derivatives were assayed for their inhibitory action on the small intestine thiamine-pyrophosphokinase activity. Their Ki values were : pyrithiamine, 2.25 X 10(-6) M; thiamine monophosphate, 4 X 10(-6) M; 2'-ethylthiamine, 8 X 10(-6) M; 2'-butylthiamine, 6 X 10(-6) M; chloroethylthiamine and dimethalium, 1.5 X 10(-5) M; amprolium, 1.8 X 10(-4) M; L-582571, 1.65 X 10(-4) M; oxythiamine, 4.2 X 10(-3) M. Of the miscellaneous compounds tested (toxopyrimidine, Na-pyrophosphate, choline, L-phenylalanine, ethyl-urethane and 5-fluorouracil), none had any inhibitory action on intestinal thiamine-pyrophosphokinase activity, even if used at concentrations hundred times higher than that of labelled thiamine.

Mechanism of binding of mouse interferon to controlled pore glass.

Many proteins bind to controlled pore glass; they are either acid elutable or alkali elutable. Mouse interferon is an acid-elutable protein. Since poly(L-lysine) and, to some extent, poly(L-arginine) are also eluted from controlled pore glass under acidic conditions, one may postulate that mouse interferon binds to controlled pore glass via some of the protein's epsilon-amino groups (of lysine) and/or guanidinium groups (of arginine) and the beads' silanol (hydroxyl groups). The necessity of lysine in the binding of interferon to controlled pore glass is further substantiated by the fact that citraconylated interferon does not bind to controlled pore glass. A requirement for Lewis acid-base interaction between the beads' B2O3 groups and the amide groups of arginine is unlikely in view of the results obtained with the alternative system, ZrOH, which, being devoid of B2O3, did bind interferon. Since a substantial amount of interferon could be eluted from controlled pore glass with ethylene glycol and high salt, one may assume that some hydrophobicity is involved in the binding of interferon to controlled pore glass.

Effect of glucocorticoid hormone on the content and synthesis of nucleic acids in cartilage of growing mice.

Glucocorticoid hormones are known to exert distinct inhibitory effects upon skeletal growth. This study examined the influence of triamcinolone hexacetonide, a long-acting synthetic analogue of cortisol, on nucleic acid synthesis in condylar cartilage of neonatal mice. It became evident that following a single injection of the hormone the DNA, RNA and protein contents were significantly reduced. The hormone inhibited both the uptake and incorporation of 3H-thymidine and 3H-uridine into DNA and RNA. Significant changes became apparent by 24 hours and persisted for 72 hours after administering the hormone. Thereafter full recovery was noted. Correlative relationship was noted between the inhibitory effects on the uptake and the subsequent incorporation of the above precursors into their respective nucleic acids. This study clearly indicates that corticosteroid hormones possess a significant inhibitory effect on the proliferative activity of neonatal chondrocytes and upon the latter's protein synthetic pathways, thereby affecting the normal process of endochondral bone growth.

[Successes, failures and complications in thrombolytic therapy in peripheral, arterial and venous occlusions].

Thrombocytic therapy in peripheral arterial and venous vessel occlusion represents a clearly described alternative towards the surgery of vessels. A success rate of 36.5% can be found in subacute peripheral arterial thrombosis and 46.3% in subacute thrombotic occlusion of a bypass-graft. Contrary to that, a rate of 29.8% can be found in complications or side-effects respectively. In cases of peripheral deep venous thrombosis, a partial or full success can be found in 72%. However, the rate of complication amounting to 44.2% is comparatively high. The longer thrombolytic therapy with streptokinase or urokinase will last, the more frequently and more serious will be the complications, such as bleedings of different kind as well as increase of temperature to mention the most frequent ones. The application of urokinase is absolutely possible today, however, the use of urokinase seems to be only justified, if a thrombolytic therapy with streptokinase was carried out successfully and a subsequent surgical therapy was not possible. The present costs of this preparation are far too high for urokinase to be applied routinely. A thrombolytic therapy with SK as well as with UK has to be followed by an anticoagulant treatment.

Electron microscope histochemistry of acetylcholinesterase distribution in the optic tectum of teleosts.

An ultrastructural analysis was made on acetylcholinesterase (AChE) localization in the optic tectum of two teleosts, the goldfish and the catfish. Electron microscope histochemistry reveals several details on synthesis, distribution and possible sites of utilization of the enzyme in the different tectal layers. The results show that AChE is synthesized by all the neuronal types present in the optic tectum. The final localization of the enzyme is the result of its synthesis in cell bodies, its storage and transport along dendrites and its release in extracellular spaces. The differences in AChE localization between the two teleosts examined mainly derive from differential enzyme release in the corresponding layers of the optic tectum. Cholinergic synapses cannot be precisely identified by means of AChE histochemistry, but the layers in which maximum release of enzyme in the extracellular spaces occurs most likely correspond to areas where cholinergic mechanisms are operating. In this connection some interesting differences of AChE localization in the superficial tectal layers (stratum marginale, stratum opticum and stratum fibrosum et griseum superficiale) are discussed. Electron microscopic histochemistry of AChE confirms its usefulness in better understanding some links between the anatomical and functional organization of complex neural structures.

Binding of exogenous DNA by human lymphocytes and by their isolated plasma membranes.

The binding of labeled bacterial DNA to human tonsil lymphocytes and to plasma membranes isolated from these cells involved macromolecules mainly located at the cell surface, since plasma membrane preparations showed properties in common with those of viable cells. DNA binding to lymphocytes was a dissociable, saturable, time, temperature, pH and concentration dependent process. A double reciprocal plot of the data obtained from DNA saturation curves gave an apparent dissociation constant of about 4 X 10(-10) M. The process of interaction of 3H-DNA with lymphocyte membranes gave a complex Scatchard plot, whose first slope yielded an apparent dissociation constant similar to that obtained for DNA-cell interaction. Binding was shown to be specific for DNA because RNA did not displace the bound DNA. Double stranded DNA was preferentially bound relative to single stranded DNA. Temperatures greater than 55 degrees C abolished the binding capacity. Results of experiments in which pH and salt concentration were varied point to the importance of conformational changes of the nucleic acid in the binding process.

Genetic polymorphism and close linkage of two alpha 1-protease inhibitors in horse serum.

Two-dimensional electrophoretic analysis of horse serum proteins was done by a first-dimension separation in agarose gel (pH 5.4) followed by a second-dimension separation in horizontal polyacrylamide gel (pH 9.0). This method resulted in improved and reproducible separation of many alpha-globulins. Two groups of alpha 1-globulins, designated Pi1 and Pi2, were found to be protease inhibitors. Preliminary studies indicated that Pi1 and Pi2 proteins differed from each other in molecular weight and in protease inhibiting spectra. Extensive polymorphism was observed for both these proteins. Family data supported the hypothesis that Pi1 and Pi2 types were controlled by autosomal codominant alleles. For both Pi1 and Pi2 systems, most of the homozygous types showed two fractions each while the heterozygous types had 4 fractions. Six Pi1 and five Pi2 alleles were observed in two breeds of Swedish horses. Complete genetic linkage was observed for Pi1 and Pi2 loci as no recombinant type was observed in 40 informative matings studied.

The assembly of lipid-linked oligosaccharides in plant and animal membranes.

Membrane preparations from growing regions of pea stems and actively-dividing mouse L-cells form lipid-linked saccharides from GDP-mannose and UDP-N-acetylglucosamine. These lipids have properties which are consistent with those of mono- and di-phosphoryl polyisoprenyl derivatives. In experiments using plant membranes, the monophosphoryl derivative labeled with GDP-(14C) mannose contains mannose only, while the diphosphoryl derivative labeled with the same nucleotide sugar is heterogeneous, containing oligosaccharides corresponding to mannosaccharides of 5, 7, and 9--12 residues. Only the diphosphoryl polyisoprenyl derivatives are labeled with UDP-(14C)glucosamine and these contain predominantly chitobiose and N-acetylglucosamine itself. Unlabeled GDP-mannose added after UDP-N-acetyl-(14C)glucosamine results in the formation of higher lipid-linked oligosaccharides which are apparently the same as those which are labeled with GDP-(14C)mannose alone. Incubation of the membranes with GDP-(14C)mannose in the presence of Mn2+, unlabeled UDP-glucose or unlabeled UDP-N-acetylglucosamine results in marked changes in the accumulation of both the polyisoprenyl monophosphoryl mannose and polyisoprenyl diphosphoryl oligosaccharides. Animal cell membranes synthesise lipid-linked oligosaccharides when incubated with UDP-N-acetylglucosamine and GDP-mannose. These oligosaccharides are similar in size to those synthesised by the plant membranes but their formation is more efficient. The potential roles of these compounds in glycoprotein biosynthesis in both plant and animal tissues is discussed.

Effects of progesterone on the oestrogen-stimulated uterus: a comparative study of the mouse, guinea pig, rabbit and sheep.

To examine more closely the anti-oestrogenic action of progesterone (P), its effect on various parameters in the 17 beta-oestradiol (E2)-primed uterus of the mouse, guinea pig, rabbit and ewe was studied. Changes in uterine wet weight, rate of in vitro protein synthesis, protein : DNA and RNA : DNA ratios, peroxidase activity and the level of cytosol receptors for E2 and P were measured. Considerable between-species differences in the effect of P on these parameters were observed. The anti-uterotrophic action was greater in the mouse than in the guinea pig and was not seen in the rabbit or ewe. P inhibited protein synthesis in the mouse, was without significant effect in the guinea pig and was mildly stimulatory in the rabbit and ewe. Inhibitory effects on protein : DNA and RNA : DNA ratios were substantial in the mouse, minor in the guinea pig and absent in the rabbit and ewe. Peroxidase activity was decrease in the mouse and guinea pig, essentially lacking in the rabbit and not detectable in the ewe. In all species the level of both oestrogen and progesterone cytosol receptors was decreased, although the effect on the E2 receptor was less marked in the ewe. The results indicate that in the species studied an effect of P on the replenishment of the E2 receptor is not necessarily associated with other anti-oestrogenic actions of P and argue against the concept that initial anti-oestrogenic actions of P are mediated via a specific effect on such replenishment.

Radioimmunoassay determinations of murine beta 2-microglobulin and an H-2 antigen-like serum component.

Normal mouse serum contains H-2 antigen-like components. One such protein consists of a 40 000 dalton chain and beta 2-microglobulin (beta 2m) and the other component contains the two H-2 antigen-like subunits and, in addition, two larger polypeptide chains with apparent molecular weights (mol. wt.) of 65 000 and 80 000. Radioimmunoassay procedures specific for the H-2 antigen-like chain and beta 2m, respectively, have been developed. Whereas free beta 2m and beta 2m associated with other polypeptide chains reacted identically in the beta 2m radioimmunoassay, the two serum forms of the H-2 antigen-like chain displayed different reactivities in the heavy-chain assay. The low mol. wt. protein complex, comprising the 40 000 dalton chain and beta 2m, inhibited in the heavy-chain assay identically with the standard. However, the high mol. wt. complex containing the H-2 antigen-like chain gave rise to a biphasic inhibition profile. This was apparently due to some antigenic determinants of the 40 000 dalton chain being buried by the 65 000 and 80 000 dalton chains. Inhibition curves parallel with the standard were, however, obtained on dissociation of the high mol. wt. protein complex with sodium dodecyl sulfate. Thus, by subjecting serum to the ionic detergent-sensitive radioimmunoassay, procedures for the two H-2 antigen-like chains could be developed. The serum levels of the H-2 antigen-like chain and of beta 2m vary with the age of the animal, and adult levels are recorded only at 6 to 7 weeks of age. The serum concentrations of the two proteins are strongly correlated. The H-2 haplotype appears to partly regulate the serum levels of the two proteins. Mice of the H-2f haplotype contain about 40-fold lower levels of the H-2 antigen-like chain than do mice of other strains.

The Lewis antigens and secretor status.

The coincidence of the presence of ABH active substances in the saliva of persons with those individuals who possess either LEWIS-d or LEWIS-b antigens on epithelial cells of their stomach tissues is examined in terms of the three-dimensional conformations of the oligosaccharide structures which form the ABH and LEWIS antigenic determinants. The H activity in saliva is normally established by examining the inhibition of the agglutination of human H red cells, which possess H (Type 2) determints at their surface, by the lectin Ulex europaeus. As was to be expected, the LEWIS-d (H-Type 1) (alpha LFuc (1 leads to 2) beta DGal (1 leads to 3) beta DGlcNAcOR, R = (CH2)8COOCH3) and LEWIS-b (alpha LFuc (1 leads to 2) beta DGal (1 leads to 3) [alpha LFuc (1 leads to 4)] beta DGlcNAcOR) determinants, obtained by chemical synthesis, do not bind Ulex. However, these structures in the de-N-acetylated form proved to do so as evidenced by their ability to inhibit the agglutination. These results together with results obtained in inhibition studies involving chemically modified H (Type 1) and H (Type 2) structures are rationalized in terms of conformational analysis and raise the possibility that the LEWIS-b and d antigens in saliva are present with the determinants in the amine form. The circumstantial evidence obtained in support of this possibility is, as yet, inconclusive. The main thrust of the paper is to indicate how modern conformational analysis may play an important role in the improvement of knowledge in carbohydrate-receptor site interaction.

Electrokinetic study of the reactions of peritoneal macrophages and eosinophils with IgG immune complexes.

Electrophoretic light scattering (laser Doppler electrophoresis) has been employed to study the effects of guinea pig IgG immune complexes on the electrophoretic mobility distributions of guinea pig resident peritoneal cells. The resident population of cells is composed of macrophages (approximately 75%) and eosinophils (approximately 25%). These cells were separated according to the well-established method of Boyum. Populations of resident macrophages, eosinophils, and the unfractionated samples were incubated with soluble immune complexes, antigen alone, or antibody alone. The mean mobility of the resident macrophages decreased approximately 60% when incubated in the presence of immune complexes, although no effect could be discerned in the presence of antigen or antibody alone. The width of the resulting macrophage mobility distribution was larger than that of the control distributions, with a broad shoulder on the high-mobility side, indicating a heterogeneous response of the macrophages to the immune complexes. Eosinophils react in two distinct fashions. One population of eosinophils is present near the control experiments. The second population reacts in a manner very similar to that of macrophages. This suggests that at least two populations of eosinophils are present in the unstimulated guinea pig peritoneal cavity. Results that are intermediate between these two cases are found when unfractionated samples are studied.

Membrane surface properties of sheep erythrocytes, an immunological reagent, after different treatments as reflected by partition in two-polymer aqueous phases.

Sheep erythrocytes (E) which, with or without certain treatments, are currently used as "immunological reagents" to detect cells with specific receptors (by rosette-formation) have been partitioned in two-polymer aqueous-phase systems selected so as to reflect charge-associated or lipid-related membrane surface properties. We have found that the partitioning behavior of E is not affected in these phases by reacting the cells with anti-E antibody (either IgG or IgM), forming EA. The additional binding of complement to the cell-antibody complex, forming EAC, results, however, in a marked decrease in the partition coefficient, K. Apparently both the charge-associated and hydrophobic properties reflected by partitioning remain accessible to the phase polymers when the cells are coated with antibody, but are not with the addition of complement. It is interesting that EA can still rosette with T-lymphocytes (14), a property of E, while the additional coating with complement results in EAC which does not appreciably do so (26). Neuraminidase or trypsin treatments of E, which yield Es having quite different rosetting properties with T-lymphocytes (14), cause increased Ks and unchanged Ks, respectively, in phases reflecting lipid-related surface properties. Either treatment causes reduced Ks of E in charged-phase systems. Neuraminidase treatment also results in a reduced electrophoretic mobility of E, while trypsin treatment is not detectable by cell electrophoresis (25). We are currently studying the possible usefulness of employing cell electrophoresis and cell partitioning in charged-phase systems jointly to obtain information on events occurring at the shear plane versus those occurring deeper in the membrane.

A millipore--blue dextran affinity binding system.

Blue dextran has been adsorbed to millipore filter discs in a linkage stable under a variety of conditions. These discs have been to bind lactic dehydrogenase, which may then be specifically eluted with NADH, one of its substrates. In contrast, another enzyme, alkaline phosphatase, not expected to bind to blue dextran, was indeed completely recovered in the filtrate.

Ca2+ is essential cofactor for trypanocidal activity of normal human serum.

Normal human serum has been known to exert a cytotoxic effect on Trypanosoma brucei subspecies for nearly 80 yr. But in spite of many attempts, no trypanocidal factor was found in human or baboon serum, until Rifkin demostrated a high density lipoprotein (HDL) in normal human serum with trypanocidal activity. The conclusion that this was the trypanocidal factor was supported by the report that serum from patients with Tangier disease, characterised by a severe deficiency of HDL, lacked trypanocidal activity. We report here that Ca2+ is an essential cofactor for the trypanocidal activity of normal human serum, in which alpha2 macroglobulin (alpha 2) might function as a Ca2+-carrier. We further show that D-glucose, D-fructose and D-mannose can suppress the trypanocidal action of normal human serum, whereas glycerol has the opposite effect.

The response of regular re-entrant supraventricular tachycardia to right heart stimulation.

The study was designed to assess the effect of various forms of right atrial or ventricular stimulation on the termination of re-entrant "supraventricular" tachycardias. Standard electrophysiological techniques were used in 81 patients to study 86 stable tachycardias. All tachycardias were initiated by single or double atrial or ventricular premature stimuli or incremental atrial pacing. Eight groups of tachycardia circuit were defined in terms of the anterograde and retrograde pathways. Termination of each tachycardia was studied by atrial underdrive, ventricular underdrive, rapid atrial stimulation and single or double atrial and ventricular premature extrastimuli. Intranodal re-entrant tachycardias formed 33% of the total and WPW tachycardias as a whole formed 55% of the total number of arrhythmias. The remainder were comprised of atrial tachycardia (5%), tachycardias in association with a partial AV nodal bypass (3%) and pre-excited tachycardias (5%). A single atrial extrastimulus was most effective where the circuit involved the right atrium. Atrial underdrive was consistently less successful than a single atrial extrastimulus in all groups. Rapid atrial pacing was effective in all groups, but caused transient atrial flutter or fibrillation in a proportion of each group except one. Ventricular underdrive stimulation was most effective in those groups where the right ventricle was involved in the circuit, but tended to be less effective than programmed single or double ventricular extrastimuli. Pacemakers designed to deliver appropriately timed single or double extrastimuli may offer an important alternative to other pacing modalities.

Joseph E. Smadel Memorial Lecture: neuroimmunologic diseases of animals and humans.

New precepts gained from the crescendo of neuroimmunobiologic research of recent decades have increased our understanding of experimental allergic encephalomyelitis (EAE), virus-associated acute disseminated encephalomyelitis (ADE), and multiple sclerosis (MS). EAE of animals and humans provides evidence of the existence in mammalian lymphoid tissues of potential clones of cells with autoreactivity for myelin basic protein (MBP) and other antigenic constituents of the central nervous system (CNS). In a new hamster model, EAE has been strikingly potentiated by persistent infection of the CNS with defective measles virus, a finding that also has implications for virus-associated ADE. Endogenous MBP or MBP degradation fragments, reactive with MBP antibodies of various affinities, have been detected by a recently devised radioimmunoassay in serum, plasma, and other body fluids of normal rats, rats with EAE, and patients with virus-associated ADE or MS. Circulating MBP or MBP fragments may be of great importance in inhibiting neuroautoimmune reactivity and play a role in repair of immunologic CNS injury should it inadvertently occur. Finally, the impressive degree of concordance of immunologic events in EAE, virus-associated ADE, and MS provides additional support for the central importance of host neuroimmunologic responses in the pathogenesis of these neutologic diseases.

The autodiagnostic pacemaker.

Loss of normal pacemaker stimulation and/or sensing functions requires prompt detection, automatic correction, and automatic and continuous "marking" of the intermittent failure. The autodiagnostic pacemaker (ADP) detects "failure to capture" (FC) by distinguishing, at its single stimulating and sensing electrode, between the normal biphasic cardiac response evoked by an adequate stimulus (corresponding to the QRS and T waves on the surface cardiogram) and the monophasic pseudo-response generated by electrotonic spread of a subthreshold stimulating current. Detection of "failure to sense" (FS) spontaneous cardiac activity requires two amplifiers: a "timing control" amplifier of standard fidelity and standard (approximately 250 ms) refractory period, and a second amplifier which has negligible refractoriness and provides high fidelity amplification of all evoked and spontaneous activity. Failure to sense (FS) is defined as a specified number of consecutive failures to recycle correctly the pacemaker's timing circuits. Similarly, a specified number of consecutive failures of the stimulus to evoke an active cardiac response is defined as a failure to capture (FC). When FC is detected, the ADP doubles the applied stimulus voltage and generates marker pulses which follow every subsequent stimulus by 40 ms. The marker pulses appear on the surface electrocardiogram, serving as an externally detectable "memory" of the earlier, possible corrected, failure. When FS is detected, non-stimulating marker pulses, of a different time relation (80 ms delay) to each stimulus, are generated continually and can also be detected externally. The ADP has been tested in 14 anesthetized, open-chest dogs. Unipolar rather than bipolar electrodes were used as they rpovided more reliable stimulation and more satisfactory electrograms for detection.

Long-term results of permanent atrioventricular sequential demand pacing.

Pervenous atrioventricular sequential demand pacemakers (AVSDPs) were implanted in 18 patients using an atrial electrode positioned in the right atrial appendage and a ventricular electrode positioned at the apex of the right ventricle. The indications included 13 patients with the sick sinus syndrome (72%), five of whom had the tachycardia-bradycardia syndrome, three with paroxysmal supraventricular tachycardia, one with cardiomyopathy and one with carotid sinus syncope. The follow-up ranged from 6 to 38 months, with a mean of 19.4 months (a total of 350 pacing months). Seventeen patients (94%) are asymptomatic. One patient had persistent episodes of paroxysmal supraventricular tachycardia. In the remaining patients with tachyarrhythmia, pacing alone (three patients) or in combination with antiarrhythmic drugs (four patients) controlled the tachyarrhythmia. There was one displacement of the atrial electrode (5.5%). Extrusion of the pacer occurred in three patients. It is concluded from this experience that AV sequential pacing is an effective technique and may be useful in patients with sick sinus syndrome, in patients with tachyarrhythmia and/or patients with poor myocardial function. However, continued research is needed to prolong battery life and to reduce the size of the pacemaker.

Automatic "scanning" by radiofrequency in the long-term electrical treatment of arrhythmias.

The use of programmed electrical stimulation in the long term treatment of re-entry tachycardia offers encouraging perspectives. Among the others proposed, the "scanning" system seems to be the most effective. However, an implantable stimulator with these features is not yet available and, thus, a temporary external lead is required. These difficulties have been overcome by utilizing radiofrequency to synchronize and stimulate. An implantable device was therefore designed which is triggered by the patient and automatically searches the interruption zone of the tachycardia by exploring the R-R cycle. The external transmitter, which can produce one or two synchronized impulses, is programmed to scan the R-R cycle with progressive steps of 5 or 10 ms; when tachycardia is interrupted, further stimulation is inhibited. The implanted module connected to an endocavitary lead does not have any power supply and, therefore, is very small. The efficacy of this method has been demonstrated in 4 patients with supraventricular tachycardia (3 with WPW syndrome) resistant to conventional pharmacologic therapy.

His bundle electrogram in patients with acute myocardial infarction.

Thirty-seven patients with acute myocardial infarction complicated by atrioventricular or bundle branch block, or a combination of both, has His bundle electrogram studies performed during their stay in the coronary care unit. The acute mortality of the 14 patients with a complicating bundle branch was 50%. Pump failure was the main cause of death. Three patients in this group had a prolonged H-V interval and one patient had a split blundle of His. The presence or absence of a prolonged H-V interval did not affect mortality in this group of patients. The acute mortality of the 16 patients with an inferior wall myocardial infarction was 6%. The H-V interval was normal in all but one of these patients. The atrioventricular block was caused by a proximal block in all cases. The acute mortality of the seven patients with an anterior wall myocardial infarction was 29%. The H-V interval was prolonged in two of seven patients. Pump failure was the acute cause of the deaths. The presence or absence of a prolonged H-V interval did not affect mortality in this group of patients.

[Family planning and demographic regression].

The problem of demographic explosion must be reexamined in the light of what has happened during the last 15-20 years in all industrialized countries, which have all experienced a sharp decrease in fertility rates due to the introduction of modern contraceptive technics, decreased mortality, and the possibility of easily obtaining induced legal abortion. Countries such as West Germany, England, Scotland, and Luxembourg have already reached zero population growth; countries such as Austria, Norway, Finland, Sweden have a growth rate of merely 0.2%. In most western countries average parity is 1.8 children per woman, when 2.18 children are needed to guarantee replacement. The sharpest decrease in fertility rates took place in West Germany and in France; should the tendency in those countries continue West Germany population would go from 60 million inhabitants to 39 million in 2030, and France would go from 53 million to 16 million. Demographic aging is another phenomenon related to fertility decrease; in 1977 in the United States 11% of the population was already over 65. Results of fertility decrease have been so sharp and so rapid that many Eastern European countries have already introduced incentives to reverse the trend, and/or to make abortion more difficult to obtain. This policy of incentives to encourage population growth has reached West Germany where since 1978 very young couples are given monetary help. Beside contraceptive availability population decrease has been fostered by the enormous increase in the rate of divorces and by the very liberal abortion laws. A large number of today's births are illegitimate, due mainly to the practice of trial marriages, without intention of real marriage, among adolescents.

Radioimmunoassay for the detection of HBeAg and anti-HBe.

We describe a solid-phase radioimmunoassay, based on the "sandwich" principle, using anti-HBe1/anti-HBe2 containing IgG precoated to polystirene beads as the solid-phase, and 125I-anti-HBE as labelled antibody for the detection of HBeAg and a two steps competitive procedure for the detection of anti-HBe. Comparison of titre obtained by ID and RIA or reference sera containing HBeAg and anti-HBe revealed that RIA is about 200 fold more sensitive than ID in detecting both HBeAg and anti-HBe. Moreover an increase in the detection rate of HBeAg from 5.9% (ID) to 15% (RIA) and of anti-HBe from 47.8% (ID) to 74.2% (RIA) and consequently a reduction of the frequency of negative samples for both HBeAg and anti-HBe from 46.3% (ID) to 10.8% (RIA) was determined by testing sera from HBsAg asymptomatic carriers. With regard to specificity, the frequency of presumptive HBeAg positive samples not confirmed was of 1 out of 29 (3.4%). Technical improvements of sensitivity and specificity of the described RIA are under development.

Clinical evaluation of the EMIT procainamide and N-acetylprocainamide assay.

Procainamide and its major metabolite, N-acetylprocainamide, were measured by the homogeneous enzyme immunoassay technique (EMIT). The reagents for the EMIT assays were supplied as a separate matched set for each assay. There is no cross-reactivity by procainamide in the assay for N-acetylprocainamide or by N-acetylprocainamide in the assay for procainamide. Within-day precision determined by replicate analysis of samples in the therapeutic range gave a coefficient of variation of less than 5% for each assay. The day-to-day coefficient of variation was less than 6% for each assay. Quantitative results obtained by the enzyme immunoassay on serum samples from patients receiving procainamide were compared with the results obtained by a high pressure liquid chromatography procedure. For the procainamide assay the correlation coefficient (r) was 0.983; for the N-acetylprocainamide assay the correlation coefficient was 0.981. There was no false positives or false negatives. The immunoassay requires 50 microliters of serum and the enzyme activity is measured in a spectrophotometer. An individual determination requires only 1 min to perform; therefore, the procedure can be used for either emergency or routine analysis.

Flow cytometry: general principles and applications to selected studies in tumor biology.

The cell populations derived from normal tissues and solid tumors comprised many different cell types. Within each cell type there is a distribution of cells in different phases of the cell cycle and/or metabolic states (ie, differing rates of protein, RNA, and other macromolecular syntheses). Flow cytometry and companion instrumentation now promise to aid in rapid quantitative analyses of heterogeneous cell populations, thus finding broad applicability in many areas of cancer research and treatment. Since it is projected that this analytical technique will greatly expend our knowledge in tumor biology, it seems appropriate to review the basis principles of the methodology and to demonstrate recent applications in several areas of current research. After reviewing basis principles, a detailed description of one specific flow cytometer, the PHYWE-ICP-22, with its computer interface as developed in this laboratory is described. Subsequently, applications of this methodology to analyses of tumor cell kinetics, assays of blastogenesis, and studies of human colon cancer are presented as specific, current applications of flow cytometry. It is anticipated that this overview of flow cytometry along with some current applications will provide a background understanding for the inevitable rapid future developments in this area of research.

Guinea-pig keratinocytes and melanocytes in tissue culture: scanning, transmission and high voltage electron microscope observations.

Keratinocytes and melanocytes cultured from guinea-pig epidermis were studied with scanning, transmission and high voltage electron microscopy to characterize the surface and internal morphology. Keratinocytes exhibited contact-inhibition and a range of surface structures consistent with cell-cycle dependent changes. Stereoscopic analysis of high voltage electron micrographs indicated regular oval nuclei with nucleoli at different depths, while thin sections revealed local channels in the nuclei. Secondary cultures differed from primary cultures in the disorder of the microfilaments, in the failure to form desmosomes, and in the failure of melanocytes to persist in culture. The beaded surface of melanocytes was indicative of underlying melanosomes that were seen in high voltage micrographs. Melanocytes were rounded with moderate ruffles or were dendritic with ruffles on the termini. These findings are discussed in relation to the observational techniques and in relation to modes of locomotion of and pigment transfer to epidermal cells.

In vivo and in vitro reactions to antigens of Haemophilus influenzae in bronchial obstruction.

The frequency of precipitating antibody to heat-labile (H(1--2) and heat-stable (HCW and HCF) antigens of Haemophilus influenzae was determined in patients with asthma, chronic bronchitis, cystic fibrosis and bronchiectasis and compared with that in a control group. This showed that the immune response of asthmatic patients to heat-stable antigens was different from that to the heat-labile antigens. Exposure to antigens of H. influenzae is common in all the disease groups. Skin test reactions having the time course and macroscopic appearance of Type I (immediate) and Type III (late) were obtained after prick and intracutaneous skin testing with HCW antigen in varying concentrations in a group of patients with asthma, chronic bronchitis or cystic fibrosis and in a control group. It is suggested that IgE and short-term sensitizing IgG antibodies may be responsible for the immediate reactions while activation of the alternative pathway of complement by endotoxin contained in HCW could be responsible for the late reactions. HCW antigens were shown to release histamine from non-sensitized human leucocytes; HCW and HCF antigens were shown to release histamine from non-sensitized human lung. None of the antigens tested had an effect on beta-receptors in tracheal preparations. It is proposed that these reactions may contribute to the pathogenicity of H. influenzae in the lower respiratory tract.

The messianic idea and messianic delusion.

The messianic delusional syndrome repeats an historical prototype that manifests itself in each patient with individual changes. The syndrome expresses a serious impairment of identity and reflects a social, cultural and religious reality through generations. The regularities of its clinical features comprise a delusional system, centered on the patient's conviction that he has been chosen by God for a special and intransferable mission. The patient has special powers for carrying out this mission. He is a savior and announces resurrection. His delusions have a clear symbolic character. For the patient's social group, the messianic idea is an attempt at annulling the effect of oppression or persecution that have become unbearable for the individual. They represent a flight from the human sphere and an attempt to be God. The patient's behavior is in consonance with this purpose; it expresses itself, on the one hand, through preaching repentance and compassion and, on the other hand, the patient gives up his earthly links and replaces them by parental relations with God. In the above-mentioned context, the author analyzes the different elements of the religious conception in the Christian, Moslem and Jewish religions, and the way each of them expresses itself in the general symptomatology.

An effect of ultraviolet light on RNA and protein synthesis in nondividing human diploid fibroblasts.

Nondividing human diploid fibroblasts maintained in medium containing 0.5% calf serum do not survive when exposed to low doses of UV (254 nm). The extent of killing is dose and strain dependent. DNA excision repair-proficient cells are more resistant than excision repair-deficient cells. Results of measurements of the effect of UV on RNA and protein synthesis in repair-proficient and -deficient (XP12BE) cells are reported. UV causes an immediate and equal depression of the RNA synthesis rate in both kinds of cells. A recovery to control rates was observed only at low (5 J/m2) doses in repair-deficient cells and at higher doses (20 J/m2) in repair-proficient cells. No recovery was observed at doses that cause substantial reductions in survival (greater than 5 J/m2 for XP12BE; greater than 40 J/m2 for repair-proficient populations). No initial effect on rate of protein synthesis was detected at doses less than 20 J/m2. However, in XP12BE populations, a decreased rate first evident at 15-30 h post-UV and before any cell degeneration and loss was observed for doses as low at 7 J/m2. This delayed effect was not observed in repair-proficient populations. The results are consistent with the hypothesis that the lethal action of UV in nondividing cells is one on DNA that leads to an inhibition of required protein synthesis by preventing RNA transcription.

Alpha-fetoprotein-associated alterations of New Zealand mouse immunopathology.

The influence(s) of chronic injections of alpha-fetoprotein (AFP), derived from murine amniotic fluid, on the natural history of immunopathology and autoantibody production in New Zealand mice was studied and compared to analagous treatment with albumin, transferrin, or phosphate-buffered saline. Treatment of young New Zealand Black (NZB) mice with AFP, at sera levels of 60-210 micrograms/ml, significantly reduces the titer of IgG1, IgG2, and IgA antierythrocyte antibodies. Similarly, such treated mice have relatively normal levels of splenic Thy-1.2-bearing cells and sera immunoglobulins, at older ages, compared to control groups. In contrast, AFP has no apparent effect on the appearance of either naturally occurring thymocytotoxic antibodies (NTa) or lymphoma. Moreover, the positive features of AFP on disease were only noted for mice treated early in life; AFP had no effect when injected into older animals. Similar, but not as dramatic, changes were observed in NZB x NZW F1 hybrids. It is concluded that in pharmacologic doses, mouse aminotic fluid enriched with AFP may alter the appearance of thymic-dependent autoantibodies.

Infants in a public school system: the indicators of early health and educational need.

The Brookline Early Education Project (BEEP) is a demonstration model in which a public school system in collaboration with a pediatric center has extended teaching and diagnostic services to the earliest days of life. In most cases, enrollment and data collection began three months before the children were born. A comprehensive diagnostic component including health, neurologic, sensory, developmental, and psychological assessments was performed periodically. The present study, the first of several reports, is an analysis of positive indicators of health and developmental need during the first six months of life. At the time of initial surveillance, age 2 weeks, a high yield of physical and demographic findings, along with perinatal stresses, was observed. It was noted that demographic, neurologic, physical, and perinatal stress factors appeared as independent variables. At the three- and six-month checkpoints, there were more overlapping findings between the categories of physical assessment, developmental examination, and neurologic evaluation. Over the six-month period, there was a tendency toward instability of findings: The group of youngsters thought to have special needs at the age of 2 weeks, 3 months, and 6 months differed in composition from each other. Likewise, there was significant flux in membership between the three- and six-month groups with needs. The study may have public policy implications, insofar as early education services should not be constructed to admit only "high-risk newborns" since this would exclude many children whose needs would become manifest later and would take in a number of children falsely identified as in need but with the resiliency to overcome this. The BEEP program has relevance for pediatric practice in demonstrating a component of health care with greater diagnostic and therapeutic responsibility for educational competence in young children.

Human tumour-associated and tumour-specific antigens: some concepts in relation to clinical oncology.

The concept of tumour-specific antigens is constantly undergoing reappraisal with the development of more sensitive methods for their detection. This has resulted in the finding that the many 'new' antigens produced by human tumours or materials immunologically closely related to them are also present in non-neoplastic tissues, albeit in small amounts. However, other antigens still appear to exist almost entirely in or on tumour cells so that the antigens of human tumours may be subdivided into either tumour-associated macromolecules or tumour-specific antigens. The elucidation of the chemical nature of the tumour-specific antigens may result in important advances in cancer diagnosis and therapy. As many are organ specific, it should be possible to evolve test systems which will enable tumours to be diagnosed and located before they become apparent clinically. On the other hand the tumour-associated macromolecules, of which the oncofetal antigens are the principal examples, are found in elevated amounts in some non-neoplastic disorders. It is now clear that serial estimation of the levels of these macromolecules is of considerably more diagnostic value than single random measurements. Current work is establishing their value in the detection of recurrent and metastatic tumours before they become apparent by other methods, which is probably their most important role, and also their value as aids to monitor therapeutic efficacy. The future use of both types of antigen may unfold a new era in cancer detection and therapy but many basic chemical and immunological studies are needed before their clinical use can be fully defined.

[Heredopathia atactica polyneuritiformis. Hexadecanoic acid storage disease (Refsum's disease). Definition, treatment and pathogenesis. A short review].

Heredopathia atactica polyneuritiformis is a clinical-genetic-biochemical entity. It is therefore not a syndrome, but a nosological entity, a morbus sui generis and it is caused by a single autosomal gene. The importance of phytanic acid accumulation in this disease was first demonstrated by Klenk and Kahlke. To the author's knowledge no case with all the typical symptoms and signs of heredopathia atactica polyneuritiformis and without any atypical clinical features, has yet been reported without a concomitant disturbance of phytanic acid metabolism. The dietary treatment--(a low phytol, low phytanic acid diet)--was first proposed by Eldjarn, when the preliminary experimental results indicated that the phytanic acid was of exogenous origin. Whatever the biochemical mechanism is between accumulation of phytanic acid and clinical manifestations in heredopathia atactica polyneuritiformis, it appears that the observations made on patients following successful dietary treatment are convincing and leave little doubt that most, if not all, the manifestations in this disease are caused in some way by the accumulation of phytanic acid.

Study of ansamycin inhibition of a ribonucleic acid-directed deoxyribonucleic acid polymerase by an immobilized template assay.

A series of structurally related ansamycins have been analyzed, in a new immobilized template assay, to determine the mechanism by which they inhibit a ribonucleic acid-directed deoxyribonucleic acid (DNA) polymerase from Moloney murine leukemia virus. By this assay, we can better correlate specific structures of these drugs with inhibitory mechanisms. Using an immobilized template, we were also able to observe drug effects on the stability of complexes formed between the polymerase, a template (polyadenylic acid-agarose), and a primer, as well as to monitor the synthesis of DNA in the presence of drug. For each drug, we determined the complex (intermediate in DNA synthesis) which was primarily affected and whether the effect was due to a destabilization process. Although the activity and specificity of the unsubstituted ansamycins (streptovaricins and rifamycin SV) were modulated by conformation of the molecule and electron density of the aromatic ring, the principal mode of inhibition is, apparently, drug binding to a polymerase-template complex; the drug binds in a manner which prevents subsequent formation of a polymerase-template-primer complex. However, some derivatives of rifamycin SV, when substituted at carbon-3 with bulky or hydrophobic side chains, displayed markedly different modes of action. For example, demethyl dimethyl rifampin prevented the formation of polymerase-template complexes, whereas rifazacyclo 16 acted by promoting the dissociation of polymerase-template-primer complexes.

Asymmetric decondensation of the L cell heterochromatin by Hoechst 33258.

A benzimidazole derivative, Hoechst 33258 can induce decondensation of constitutive heterochromatin in the mouse derived L cell chromosomes when the compound is given in sufficiently high concentration (40 micrograms/ml) to the L cell culture. Hoechst 33258 at low concentration (1 micrograms/ml, 16 h) cannot produce this effect on L cell chromosomes. Bromodeoxyuridine (BUdR) incorporation for one cell cycle simultaneous with the Hoechst 33258 treatment at low concentration could decondense heterochromatin segments in metaphase chromosomes. The heterochromatin decondensation, however, was asymmetric; it was observed only on one chromatid and the other of a chromosome remained in condensed state. The observation of asymmetric decondensation of heterochromatin by Hoechst 33258 after BUdR incorporation for one cell cycle, the association of A-T rich satellite DNA to mouse heterochromatin, and available data on the specific binding of Hoechst 33258 to A-T base pairs of DNA and on the higher affinity of the compound to BUdR substituted DNA than to ordinary DNA implied that the binding of Hoechst 33258 molecules to A-T rich satellite DNA is the cause of heterochromatin decondensation.

Further evidence for inhibition of episodic luteinizing hormone release in ovariectomized rats by stimulation of dopamine receptors.

Stimulation of dopamine receptors by apomorphine inhibits episodic LH release in ovariectomized rats. The present study was designed to examine further the role of dopamine in this process. Unrestrained, unanesthetized rats with indwelling right atrial cannulae were bled continuously (30 or 50 microliters of whole blood/5 min for 3-6 h) and whole blood samples analyzed for LH by radioimmunoassay. Animals were treated with various compounds reported to stimulate or block dopamine receptors. ET 495, a long acting dopamine receptor stimulating agent, caused a marked inhibition of episodic LH release (2 1/2-4 h). Control injections of distilled water had no effect. d-Butaclamol, a blocker of dopamine receptors, did not itself alter episodic LH release but prevented the inhibitory effects seen following apomorphine or ET 495. I-butaclamol, a biologically inactive form of butaclamol, had no effect. Measurement of plasma corticosterone levels in these same animals indicated increased values following apomorphine or ET 495 alone (when LH release was inhibited), as well as after apomorphine or ET 495 administration to d-butaclamol-pretreated rats (when LH levels did not change). These data support our previous hypothesis that in ovariectomized adult rats, activation of dopamine receptors is capable of inhibiting episodic LH release, but that dopamine may not play an inhibitory role under normal physiological conditions in the modulation of LH secretion. In addition, the inhibitory action of apomorphine and ET 495 does not appear to be exerted via a stress-induced release of adrenal corticosterone.

Television epilepsy--the role of pattern.

Patients with photosensitive epilepsy were asked to view normally functioning 625-line televisions while the EEG was monitored. In the first of two studies paroxysmal EEG activity was reliably induced by television at a viewing distance related to a patient's sensitivity to intermittent photic stimulation (IPS); patients who were sensitive to diffuse IPS at 50 Hz were sensitive to the television at greater viewing distances than those who were not. No such relationship was obtained with patterned IPS. On the other hand, patterned IPS was generally more epileptogenic than diffuse IPS with the same luminance. In the second study, where the angular subtense of the television screen and the subtense of its lines were manipulated independently, the convulsive response was found to be a function of both factors, the relative contribution of each depending on the viewing distance at which the patient was sensitive. For patients sensitive at normal viewing distances, where 50 Hz diffuse flicker appeared to be responsible for the induction of paroxysmal activity, the probability with which paroxysmal activity was induced was closely related to the subtense of the screen. For patients sensitive only at closer viewing distances the probability was influenced not by the subtense of the screen but by the subtense of its lines, suggesting that the paroxysmal activity was induced by the 25 Hz pattern alternation produced by the scan. A television with a small screen was considerably less epileptogenic than one with a large screen for all patients, presumably due to the reduced contribution of both diffuse flicker and pattern alternation.

The measurement of receptive field properties of neurones in the visual cortex of the conscious, mobile cat.

We have assessed the accuracy, reliability and ease of use of a technique for determining the receptive field properties of cells in the visual cortex of the conscious, unrestrained cat. Our tests have provided the following results: (1) Minimal behavioural training is necessary; it is often possible to train an animal in only two sessions. (2) A surprising degree of optical stability can be achieved. Measurements of receptive field properties are consequently reliable and repeatable. (3) We are not certain of the exact limits of accuracy which can be achieved, but we usually determine orientation specificity +/- 5 degrees, and measure receptive field positions +/- 1.5 degrees.

Impaired sphincter control in the dyspnoeic.

6 case-histories illustrate how altered control of micturition is an occasional accompaniment of dyspnoea in patients aged over 50. Recognition of this problem can be a step towards understanding the great disability of some dyspnoeic patients.

Microspectrophotometric detection of heparin in young and adult rat mast cells, human mast cells and human basophilic granulocytes stained metachromatically with toluidine blue O.

A qualitative microspectrophotometric detection method for heparin in situ has been developed, using data obtained previously with a model system of polyacrylamide films containing pure glycosaminoglycans (Tas & Roozemond 1973, and Tas 1975). This technique, based on the unique metachromatic properties of heparin with Toluidine Blue O in glycerol, has been worked out with rat peritoneal and mesenteric mast cells. After equilibration of the stained, air-dried cells in glycerol (for some days), the specific peak of the heparin-Toluidine blue O complex (as found in the model experiments at about 515 nm) could be recorded. It was found that the method can be used to detect unequivocally the presence of heparin in cells, even if they also contain up to 75 mole percent of other, lower sulphated-glycosaminoglycan. Otherwise the method might yield some information about the degree of sulphation of the heparin concerned. With this technique the presence of heparin has been proved in young and adult rat mast cells and for the first time now directly in normal human mast cells and normal human basophilic granulocytes.

Determination of the human lymphokine leukocyte migration inhibitory factor (LIF) by a sensitive radioenzymatic assay. Inhibitory effect of cGMP on the esterolytic activity of highly purified LIF.

Indirect experiments using irreversible enzyme inhibitors have shown that the human lymphokine leukocyte migration inhibitory factor (LIF) is a serine esterase and protease exhibiting specific affinity towards arginine esters and amides. A sensitive assay for direct measurement of esterase activity using p-tosyl-L-arginine (3H) methyl ester (3H-TAME) as substrate is described. Esterolytic activities are demonstrated in crude supernatants of human lymphocytes stimulated with concanavalin A (LIF-rich) and, more pronounced, in supernatants of unstimulated cells (control). To follow the effects of purification procedures, the serine esterases of LIF-rich and control preparations were specifically labeled with the irreversible, active site directed agent (1,3-3H)di-isopropylphosphorofluoridate. Most of these enzymes, visualized by Sephadex chromatography, were removed by a gentle three-step procedure, allowing at least 50% of the initial LIF activity to be recovered. The resulting LIF-rich preparation, purified to contain serine esterases at a concentration corresponding to less than 1 ng per ml original supernatant, still showed estrolytic activity towards 3H-TAME. The optimal conditions for the radioenzymatic assay of purified LIF and the inhibitory effect of 10(-4) M cGMP, which on the basis of indirect experiments has been implicated as a specific regulator of LIF activity, are described.

[The fatty acid composition of the gangliosides in different parts of the vertebrate brain].

Comparative studies on gangliosides from the brain of higher (mammals, birds) and lower (amphibians, cartilaginous and teleost fishes) vertebrates indicate that for each of the parts of the brain the same regularities in changes of fatty acid composition are typical as for the total brain in a series of vertebrates. All brain structures of the mammalian brain exhibit higher saturation of ganglioside fatty acids, higher content of stearic acid (C18:0) and lower relative content of long chain (C22-C24) fatty acids, especially that of C24:1, as compared to the corresponding brain structures of lower vertebrates. Fatty acid composition in birds is more close to mammalian one with respect to some of the features, while with respect to the other ones it occupies an intermediate position between lower vertebrates and mammals. In the series more phylogenetically ancient--more recent brain parts (medulla oblongata--midbrain--forebrain), the content of content of stearic acid and the degree of saturation ganglioside fatty acids increase, whereas the relative content of acids with a long chain (C22-C24) in them decreases (with the exception of amphibians). In all the brain structures of higher elasmobranch fishes, fatty acid composition of gangliosides exhibits more progressive features of chemical organization as compared to that in homologous structures of lower elasmobranch fishes.

Aortic pulse wave velocity, elasticity, and composition in a nonhuman primate model of atherosclerosis.

Aortic pulse wave velocity was determined in Macaca fascicularis monkeys fed either atherogenic or control diets for 36 months. The foot-to-foot velocity and apparent phase velocities of the second through seventh Fourier harmonics at a given diastolic pressure in the atherosclerotic monkeys were 1.5 to 2.0 times the values for the control animals. More than 80% of the aortic intimal surface of the atherosclerotic monkeys was covered with fibrous or fatty plaque, which approximately doubled wall thickness and wall thickness to radius ratio. Angiochemical evaluations showed no difference in collagen or elastin concentration (as a fraction of lipid and mineral-free dried aorta), but the atherosclerotic aortas were 1.5 to 2.0 times that of control in collagen and elastin content (defined as the absolute quantity beneath a square centimeter of intimal surface). Total cholesterol and calcium concentrations in the atherosclerotic aortas were more than 10 times the values for the control aortas. The static circumferential distensibility of the excised atherosclerotic aortas was significantly less than control, but there was no difference in incremental (Young's) modulus of elasticity. The in vitro pressure-strain elastic modulus of the atherosclerotic aortas was more than twice that of control, which was predicted from the enhanced wave velocity. The significantly increased stiffness of the atherosclerotic arteries appeared to be due mainly to the increased wall thickness caused by the atherosclerotic plaques rather than to material changes described by Young's modulus. Extensive medial damage, however, also was present and could have had a major influence on stiffness. Atherosclerosis therefore can result in increased aortic stiffening, detectable by pulse wave velocity, even if there is no change in the overall Young's modulus of elasticity.

Functional morphology of forelimb joints in the woolly monkey Lagothrix lagothricha.

This gross anatomical study of embalmed forelimb joints of the South American woolly monkey Lagothrix lagothricha focuses on the problem of determining in osteoligamentous preparations how the disposition of the capsular apparatus and the geometry of the articular surfaces govern the amount and types of movement permitted at a joint, and then correlates these findings with the use of the forelimb in the positional capabilities of captive wooly monkeys observed by this author. Data collection was by dissection, quantitative range of motion studies on osteoligamentous preparations, and by qualitative manipulations of these preparations and of disarticulated bones. Supplemental evidence was obtained from radiographs and from an estimate of angular values of articular surfaces. Presented for the shoulder, elbow, radioulnar, and hand joint complexes are the functional anatomy of the capsular apparatus and articular surfaces, the quantitative range of motion data, and proposed mechanisms of movement that combine the functional morphology with the observed use of the forelimb in activities by the animal. The structurofunctional framework of MacConaill is the basis for the joint analysis. The evidence suggests that the functional anatomy of these joints correlates well with the possible positional capabilities of Lagothrix. The morphology of the capsular apparatus and joint surfaces reflect both the arboreal quadrupedalism and forelimb suspension of the woolly monkey. The positions of maximum congruency and close-pack approximate the forelimb's weight-bearing stance during palmigrade arboreal quadrupedalism. During forelimb suspension, the taut and twisted capsular apparatus of close-packed joints maintains the integrity of forelimb links in tension. This is the first known report of the internal band of the collateral ligament of the proximal interphalangeal joints of the second through fifth digits being incorporated into the terminal tendon of the extensor assembly. This author believes this is a mechanism not only for maintaining the integrity of these joints during forelimb suspensory activities when the body is supported by the middle phalanges, but also for helping to coordinate the flexion-extension actions of the phalanges for smooth grasping and release maneuvers in an arboreal environment. When sufficient comparative data have been collected, such correlations of the functional morphology of joints with positional behavior may be used to postulate positional capabilities of fossil primates.

Murine heavy chain disease.

Using cultured mouse myeloma cells, it has been possible to derive cells which are now synthesizing products similar to human heavy chain disease proteins. An initial mutant was isolated which snythesized a heavy chain with an internal deletion and a normal light chain. In a subsequent step, a variant was identified in which the synthesis of the heavy chain with the deletion persisted in the absence of light chain synthesis. These experiments also demonstrate that in the MPC-11-derived cultured cell line, the continued synthesis of heavy chain does not require the synthesis of light chain.

Mechanisms of neural integration in the parietal lobe for visual attention.

The impulse discharges of neurons in the inferior parietal association cortex (area 7) were studied in the alert, behaving rhesus monkey, trained to fixate and follow visual targets. Four classes of cells related to visual or visuomotor function were found. Cells of one of these are sensitive to visual stimuli and have large, contralateral receptive fields with maximal sensitivity in the far temporal quadrants. Cells of the other three classes are related to visuomotor functions: visual fixation, tracking, and saccades. They are neither sensory nor motor in the usual sense for they are activated only by interested fixation of gaze or tracking, or before visually evoked saccadic eye movements. They are not activated during the spontaneous saccades and fixations that the monkey makes while casually exploring his environment. It is hypothesized that the light-sensitive neurons provide the visual input to the visuomotor cells that, in turn, produce a command signal for the direction of visual attention and for shifting the focus of attention from one target to another.

[Transplantation of isolated islands of Langerhans for the treatment of diabetes mellitus].

The historical development of research on islets transplantation is briefly reviewed followed by a description of a technique using Ficoll gradient separation to obtain islets of Langerhans from the rat pancreas. It was possible to increase the number of islets obtained with this procedure by modifying it in several ways. The islets were then transplanted in an isologue manner into rats with streptocotocin-induced diabetes. The effect of transplantation on glucose metabolism in these rats was evaluated both by determination of the general symptoms typical for diabetes as well as by the performance of such functional tests as IVGTT, GAC and tolbutamide test. The islets could be transplanted into various locations, but it was found that positive results were obtained only if the liver was the site of application and if than 2100 islets were used. Three hours after transplantation normalization of blood sugar levels and serum insulin could be observed; these levels remained stable over 18 months. It was possible to transplant in the same animal several times. This had an effect on the metabolism which was equivalent to the sum of the separate transplants. By means of both light and electron microscope examination the morphological changes which the transplanted islets underwent at the site of transplantation were observed.

Viability and endogenous substrates used during starvation survival of Rhodospirillum rubrum.

Cells of Rhodospirillum rubrum were grown photoorganotrophically and chemoorganotrophically and then starved for organic carbon and combined nitrogen under four conditions: anaerobically in the light and dark and aerobically in the light and dark. Illumination prolonged viability and suppressed the net degradation of cell material of phototrophically grown cells, but had no effect on chemotrophically grown cells that did not contain bacteriochlorophyll. The half-life survival times of carbohydrate-rich phototrophically grown cells during starvation anaerobically or aerobically in the light were 17 and 14.5 days, respectively. The values for starvation aerobically and anaerobically in the dark were 3 and 0.5 days, respectively. Chemotrophically grown cells had half-life survival times of 3 and 4 days during starvation aerobically in the light and dark, respectively, and 0.8 day during starvation anaerobically in the light or dark. Of all cell constituents examined, carbohydrate was most extensively degraded during starvation, although the rate of degradation was slowest for phototrophically grown cells starved anaerobically in the light. Phototrophically grown cells containing poly-beta-hydroxybutyrate as carbon reserve were less able to survive starvation anaerobically in the light than were carbohydrate-rich cells starved under comparable conditions. Light intensity had a significant effect on viability of phototrophically grown cells starving anaerobically. At light intensities of 320 to 650 lx, the half-life survival times were 17 to 24 days. At 2,950 to 10,500 lx, the survival times decreased to 1.5 to 5.5 days. The kinetics of cell death correlated well with the rate of loss of cell mass of starving cells. However, the cause of death could not be attributed to degradation of any specific cell component.

Molybdenum independence of nitrogenase component synthesis in the non-heterocystous cyanobacterium Plectonema.

The cyanobacterium Plectonema boryanum (IU 594-UTEX 594) fixes N2 only in the absence of combined N and of O2. We induced nitrogenase by transfer to anaerobic N-free medium and studied the effect of Mo starvation on nitrogenase activity and synthesis. Activity was first detected within 3 h after transfer by the acetylene reduction assay in controls, increasing for at least 25 h. Cells grown on nitrate and Mo and then transferred to N-free, Mo-free medium produced 8% of the control nitrogenase activity. Addition of W to the Mo-free medium reduced the activity to 0.5%. Under both Mo starvation conditions, nitrogenase protein components were synthesized. Component II of the cyanobacterial enzyme was detected by in vitro complementation with Mo-containing component I from Klebsiella pneumoniae or Azotobacter vinelandii but not Clostridium pasteurianum. Component I activity was restored by addition of Mo to cultures in which new enzyme synthesis was blocked by chloramphenicol. Acidified extracts of Plectonema induced in Mo-containing medium contained the Fe-Mo cofactor required to activate extracts of the Azotobacter mutant UW45 in vitro, but they did not activate extracts of Mo-starved Plectonema. Analysis of 35SO4(2-)-labeled proteins by polyacrylamide gel electrophoresis suggested that Mo is required for the conversion of a high-molecular-weight precursor to component I in Plectonema.

Polyamines of Anacystis nidulans and metabolism of exogenous spermidine and spermine.

Several biochemical parameters, including that of polyamine content, accompanying the growth of the cyanobacterium Anacystis nidulans were studied. At all stages of growth under autotrophic conditions, the organisms were found to be rich in spermidine and lacking in spermine, as is typical of procaryotic organisms. The cells were quite low in putrescine, and no unusual polyamine was observed to be present as a major component. Conjugated polyamines were not detected in the cultures. At maximal culture density, the levels of spermidine, DNA, RNA, protein, and chlorophyll were also maximal. Shortly after the inception of the stationary phase, the spermidine content of the cells was the first parameter observed to decrease in cultures which were shortly to become yellow. Spermidine lost from the cells was not recovered in the medium in a free or conjugated form. This indication of degradation of spermidine was studied by the addition of polyamines to growing cultures. Exogenous spermidine and spermine were found to be metabolized rapidly by the organisms, of which diaminopropane was one product. Putrescine was found to be markedly toxic, whereas spermidine, some other triamines, and spermine were much less toxic.

Biochemical studies of rat liver Golgi apparatus. I. Isolation and preliminary characterization.

A method is described for the isolation of morphologically well-preserved Golgi apparatus from rat liver. The method is essentially the same as that of Morré et al. (Morré, D.J., Hamilton, R.L., Mollenhauser, H.H., Mahley, R.W., Cunningham, W.P., Cheetham, R.D., & Lequire, V.S. (1970) J. Cell Biol. 44, 484-491) except that mild cell disruption is achieved by means of a stainless-steel sieve. The average recoveries of protein and galactosyltransferase in the isolated fraction are about 6 mg from 10 g of perfused liver and about 35% from the homogenate, respectively. The preparation is virtually free from succinate-cytochrome c reductase, glucose-6-phosphatase, acid phosphatase, and 5'-nucleotidase. The Golgi fraction as well as its vesicular fragments is homogeneous upon isopycnic centrifugation in both sucrose and dextran density gradients. Their buoyant densities in sucrose are significantly higher than those in dextran, indicating that both forms of the organelle are closed systems which are impermeable to macromolecules. The galactosyltransferase activity of a freshly prepared Golgi fraction, measured with ovalbumin as galactosyl acceptor, is activated 26-fold by the addition of Triton X-100, whereas those of homogenized, sonicated, and aged preparations are only activated 2- to 4-fold.

Biochemical and immunological characterization of human opsonic alpha2SB glycoprotein: its identity with cold-insoluble globulin.

The relationship between human cold-insoluble globulin (CIg, plasma fibronectin) and the human serum opsonic alpha2SB glycoprotein was investigated using immunochemical and biochemical techniques. The two proteins appeared to have identical molecular weights by sodium dodecyl sulfate-polyacrylamide gel electrophoresis on 3.3% gels; have identical migration in the native state on 2.7 to 27% gradient polyacrylamide gels; and have a similar amino acid composition within the accuracy of analysis. Human serum demonstrates antigenic identity when diffused against monospecific antisera to both proteins confirming the presence of common antigenic sites on both molecules. Purified human serum opsonic alpha2SB glycoprotein and purified CIg also demonstrate antigenic identity when diffused against monospecific antiserum to either of the isolated proteins. Antiserum to both proteins also inhibits in vitro hepatic Kupffer cell phagocytic uptake of test particles. These results suggest the idenity of these two proteins and reveal a major physiological function for human plasma CIg. Thus, CIg may be important in the regulation of hepatic reticuloendothelial phagocytic activity and nonspecific systemic host defense. This process of systemic host defense has been shown to be depressed in patients following trauma, major surgery, burn injury, and during neoplastic disease, and, in part, mediated by a deficiency or depletion of the alpha2SB glycoprotein.

Value of acid metabolic products in identification of certain corynebacteria.

Acid metabolic products of 23 strains of human and animal pathogenic corynebacteria, representing eight different species, were determined by gas chromatography. The results showed that the species examined were metabolically heterogeneous and could be presumptively identified based on the acid products produced. Corynebacterium equi did not produce any acids; C. renale produced lactate; and C. pyogenes produced major amounts of lactate, variable amounts of acetate, and minor amounts of succinate and pyruvate. C. kutscheri produced propionate and lactate as major products and pyruvate and oxalacetate as minor products. C. diphtheriae and C. pseudotuberculosis produced major amounts of propionate, acetate, and formate. In addition, C. pseudotuberculosis produced major amounts of pyruvate and minor amounts of succinate, lactate, and oxalacetate, whereas C. diphtheriae strains produced minor but variable amounts of lactate, succinate, fumarate, pyruvate, and oxalacetate. C. bovis produced aicd products similar to those of C. pyogenes but was readily distinguishable from the latter by the lack of hemolysis on blood agar, colony morphology, catalase reaction, and biochemicals. C. suis characteristically produced major amounts of ethanol, acetate, and formate and minor amounts of lactate and succinate but no propionate.

Parathyroid hormone responses to catecholamines and to changes of extracellular calcium in cows.

Modifications of the plasma level of immunoreactive parathyroid hormone (PTH) in cattle were induced by changes of the plasma concentrations of epinephrine, isoproterenol, or calcium. During abrupt hypocalcemia, PTH, obtained by infusions with ethylene glycol-bis (beta-aminoethylether) N, N'-tetraacetate (EGTA), increased during the first 4-8 min. After a transient decline, the hormone levels rose again and remained elevated. Infusions of calcium suppressed the hypocalcemia-induced augmentation of PTH levels within a few minutes. Prolonged epinephrine (and isoproterenol) infusions also rapidly increased PTH levels, however, in this case, they returned to basal concentrations after 50-60 min. Additional epinephrine infusions could not further raise PTH values. Moreover, three short-lasting infusions of epinephrine (7 min each), given at 30-min intervals, increased PTH levels to the same extent, whereas additional infusions were much less effective. The PTH response to epinephrine was completely restored, when the interval after a prolonged epinephrine infusion had been prolonged to > 100 min. During moderate hypocalcemia, occurring at the end of EGTA infusions and lasting for 90 min, the PTH response to a short-lasting epinephrine infusion (7 min) was more pronounced than in normocalcemic animals. During severe hypocalcemia, in which superimposed short-lasting infusions of EGTA (7 min) led to an additional abrupt fall of plasma calcium concentrations but not to a corresponding additional rise of the PTH levels, epinephrine rapidly and further increased PTH concentrations. On the other hand, at the end of prolonged infusions of epinephrine, when additional infusions of epinephrine were ineffective in raising PTH levels, EGTA-induced hypocalcemia consistently increased PTH concentrations. The EGTA-induced augmentation of PTH levels was enhanced by epinephrine and isoproterenol but not by propranolol. The present findings indicate, that variations of the extracellular calcium concentrations and beta-adrenergic agonists modify PTH levels by two different and independent mechanisms. On the other hand, it appears that the magnitude of change of the PTH levels to either stimulus is partially modulated by exposure to the other.

The kallikrein-kinin system in Bartter's syndrome and its response to prostaglandin synthetase inhibition.

The kallikrein-kinin system was characterized in seven patients with Bartter's syndrome on constant metabolic regimens before, during, and after treatment with prostaglandin synthetase inhibitors. Patients with Bartter's syndrome had high values for plasma bradykinin, plasma renin activity (PRA), urinary kallikrein, urinary immunoreactive prostaglandin E excretion, and urinary aldosterone; urinary kinins were subnormal and plasma prekallikrein was normal. Treatment with indomethacin or ibuprofen which decreased urinary immunoreactive prostaglandin E excretion by 67%, decreased mean PRA (patients recumbent) from 17.3+/-5.3 (S.E.M.) ng/ml per h to 3.3+/-1.1 ng/ml per h, mean plasma bradykinin (patients recumbent) from 15.4+/-4.4 ng/ml to 3.9+/-0.9 ng/ml, mean urinary kallikrein excretion from 24.8+/-3.2 tosyl-arginine-methyl ester units (TU)/day to 12.4+/-2.0 TU/day, but increased mean urinary kinin excretion from 3.8+/-1.3 mug/day to 8.5+/-2.5 mug/day. Plasma prekallikrein remained unchanged at 1.4 TU/ml. Thus, with prostaglandin synthetase inhibition, values for urinary kallikrein and kinin and plasma bradykinin returned to normal pari passu with changes in PRA, in aldosterone, and in prostaglandin E. The results suggest that, in Bartter's syndrome, prostaglandins mediate the low urinary kinins and the high plasma bradykinin, and that urinary kallikrein, which is aldosterone dependent, does not control kinin excretion. The high plasma bradykinin may be a cause of the pressor hyporesponsiveness to angiotensin II which characterizes the syndrome.

Immunoglobulin secretion by human splenic lymphocytes in vitro: the effects of antibodies to IgM and IgD.

The effects of F(ab')2 fragments of affinity-purified rabbit anti-human mu chain antibody (RaHmu) and rabbit anti-human delta chain antibody (RaHdelta) on spontaneous and mitogen-stimulated immunoglobulin (Ig) secretion by normal human spleen cells were studied. IgM and IgG secretion by human spleen cells cultured in vitro was measured by incubating the cells with 3H-amino acids precipitating the secreted labeled Ig with anti-Ig, and analyzing the precipitates by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Both RaHmu and RaHdelta suppressed spontaneous and LPS-induced IgM and IgG secretion as well as PWM-stimulated IgG secretion. In different experiments, RaHmu and RaHdelta either suppressed or augmented PWM-induced IgM secretion. The anti-Ig induced augmentation of PWM-triggered IgM secretion was most apparent when spleen cells were cultured at lower cell densities or when lower concentrations of anti-Ig were employed. These date indicate that perturbation of B cell surface immunoglobulin receptors with specific anti-Ig antibody can alter markedly the ability of these cells to differentiate into antibody-secreting cells.

Isolated T wave alternans elicited by hypocalcemia in dogs.

Isolated T wave alternans (TWA) could be consistently induced in 13 dogs by infusion of an acid dextrose solution (ACD) and in nine with a 2% EDTA disodium solution. To study the factors concerned in evoking TWA, serial determinations of total and ionized serum calcium (ICa), Mg, K, pH, pCO2 and pO2 were performed. Arterial or LV pressures and LV dp/dt as well as a standard electrocardiographic lead were monitored. TWA appeared after the QT interval was significantly prolonged and was associated with a fall in LV pressure and with mechanical alternans. QT alternans could also be detected in 50% of the cases. ICa levels were significantly reduced at the moment of appearance of the TWA. Data obtained support the conclusion that TWA was the result of a reduction in the level of the ICa, probably interfering with the calcium transport mechanisms of the myocardial cell, although the possibility that the alternans may be due to other effects of the two agents employed unrelated to calcium lowering cannot be excluded.

Effect of load disturbances during centrally initiated movements.

1. We have investigated the relative contributions of mechanical and reflex mechanisms in generating the forces produced by the neck muscles when loads were unexpectedly applied during centrally programmed head movements in monkeys. These movements, subserved by muscles well endowed with muscle spindles, are part of the coordinated eye-head response to the appearance of a stimulus in the animal's visual field. Our preparation was a chronically vestibulectomized monkey trained to make a visual discrimination. 2. Two procedures were used to evaluate the torque generated by the neck musculature when an unexpected load disturbance was applied: first, by surgically interrupting the afferent loop subserving the reflex action (section of cervical dorsal roots) and second, by building a mathematical model of the head-neck system and carrying out a process of simulation. 3. Our results indicated that the compensatory torque of reflex origin stimulated by the application of an opposing force was less than 10--30% of that required for perfect compensation, and the larger fraction of the observed compensation was due to the mechanical properties (inertial, viscous, and elastic) of the neck musculature. The combined action of reflex and mechanical processes never completely compensated for the disturbance.

Role of primate flocculus during rapid behavioral modification of vestibuloocular reflex. II. Mossy fiber firing patterns during horizontal head rotation and eye movement.

1. Extracellular recordings were obtained from 113 mossu fibers (MFs) in the flocculus of alert monkeys trained to perform a visual tracking task during sinusoidal, horizontal head rotation. The analysis of MF discharge patterns was designed to allow quantitative comparison of the discharge properties of flocculus MFs with brain stem cell populations from which the MFs might originate and with flocculus Purkinje cells (P-cells). Based on their firing patterns, MFs were divided into two classes. Vestibular MFs discharged in relation to head velocity and, in some cases, also in relation to eye movement. Eye movement MFs discharged only in relation to one or more components of eye movement. 2. Vestibular MFs were subdivided into three classes. Vestibular-only MFs (n = 15) displayed a modulation in firing rate during head rotation but exhibited no relationship to spontaneous eye movements. Vestibular-plus-saccade MFs (n = 14) displayed a modulation in firing rate during head rotation that quantitatively resembled the modulation in vestibular-only MFs. In addition, a pause in firing rate interrupted the vestibular modulation during saccades in one or more directions. Vestibular-plus-position MFs (n = 4) exhibited steady firing rates that were linearly related to horizontal eye position in the absence of vestibular stimulation. Sinusoidal head rotation evoked a modulation ofiring rate above and below the firing rate set by the eye position. 3. during sinusoidal head rotation, vestibular MF firing rate led head velocity by an average of 24 degrees. The amplitude of MF firing-rate modulation increased as a function of the frequency of head rotation and, hence, maximum head velocity. Since these characteristics are similar to those displayed by P-cells during suppression of the VOR, vestibular MFs probably transmit the head velocity component of P-cell firing rate to the flocculus. Based on evidence from other mammals and a quantitative comparison of population discharge characteristics, it is likely that vestibular MFs originate from the vestibular nerve and from cells in the medial vestibular nucleus. 4. Based on their discharge patterns, eye movement MFs were also subdivided into three classes. Burst MFs (n = 14) emitted a high-frequency burst of spikes prior to and during saccades in one or more direction, but were silent during steady fixation. Burst-tonic MFs (n = 53) emitted a burst of spikes prior to saccades in a preferred ("on") direction, ceased firing during saccades in the opposite ("off") direction, and exhibited steady firing rates that increased as steady gaze shifted in the on direction. Tonic MFs (n = 13) displayed steady firing rates that increased as the position of steady gaze shifted in the on direction, and either paused or exhibited step changes in firing rate during saccades. 5. During steady fixation, 64% of tonic and burst-tonic MFs were recruited into maintained firing within +/- 10 degrees of the primary direction of gaze...

L-alanyl-L-tyrosine as a tyrosine source during intravenous nutrition of the rat.

Tyrosine is considered an essential amino acid for some premature infants, but its limited solubility prevents addition to intravenous solutions in adequate amounts. Tyrosine peptides with solubilities greater than tyrosine were synthesized to evaluate potential for intravenous use. Intraperitoneal injection of L-alanyl-L-[U-14C]tyrosine (0.5 mmoles/kg) into adult rats resulted in a rapid labeling of tissue pools and production of 14C-CO2. When the peptide was infused as part of an intravenous nutrition solution (0.5 mmoles/kg/24 hours) into adult rats, plasma and tissue tyrosine pools were rapidly labeled. The radioactivity distribution after 24 hours of infusion was: 41% CO2, 13.4% muscle, 7.7% urine, 7.1% liver, 5.5% intestine, 1.4% kidney, with the remainder in other carcass organs. No unhydrolyzed peptide was found in tissue homogenates. Between 9% and 17% of the radioactivity present in tissue was free tyrosine, with the remainder in tissue protein. Tyrosine accounted for more than 96% of this protein bound radioactivity. The data indicate good utilization of alanyl-tyrosine as a tyrosine source when administered intravenously.

Hemodynamic effects of systemic and central administration of alpha-methyldopa.

Systemic and regional hemodynamic changes were measured in five restained conscious rhesus monkeys before and after a 2-hour intravenous infusion of 50 mg/kg of alpha-methyldopa and, in another group of five monkeys, 5 to 10 mg injected into a lateral cerebral ventricle. Both routes of administration evoked similar degrees of hypotension, bradycardia and decreased cardiac output, although the cerebral intraventricular (i.c.v.) injections had more immediate and long-lasting effects. Both groups had a similar pattern of changes in the redistribution of cardiac output and blood flow that lasted at least 4 hours. Blood flow was maintained in the hepatic and renal arteries and decreased in skeletal muscle, heart, brain and skin. In contrast, i.c.v. injections of alpha-methyldopamine and alpha-methylnorepinephrine given at the same site evoked dose-related pressor responses that lasted up to 4 hours. The data suggest that alpha-methyldopa has important central action that inhibits sympathetic outflow, but that its hypotensive effect is either mediated only by endogenously formed metabolites or that its mechanism of action is not directly related to these metabolites at sites around the lateral and third cerebral ventricles in the monkey.

Cellular changes in the toad urinary bladder in response to metabolic acidosis.

The urinary bladder of Bufo marinus excretes H+ and NH+4, and the H+ excretion is increased when the animal is placed in metabolic acidosis. The mitochondria-rich (MR) cells mediate the H+ excretion by the bladder. The purpose of this study was to determine if there is a change in MR cells of the bladder during metabolic acidosis. Bladders from normal toads and from toads that had been placed in metabolic acidosis were used. The bladders were mounted between plastic chambers and H+ excretion measured. The bladder was then fixed and prepared for scanning (SEM) and transmission (TEM) electron micrograph studies. SEM's at low magnification were used to count the various cell types and the TEM's were used to confirm the different cell types. Fields were randomly selected and a total of 2500 cells counted in each group. The bladders from toads in metabolic acidosis had a consistently higher ratio of MR cells to granular cell than did the normal bladders. These results indicate that during metabolic acidosis there is an increased number of MR cells in the bladder, and this increased the bladder's capacity to excrete H+.

[The immediate reconstruction of a large lip-chin defect with two muscular pedicled and innervated island flaps (author's transl)].

Different methods of defect closure in the lipchin region through lateral advancement-flaps have been described by Bernard, Fries, Hertig, Meyer and others. Is there not enough material available, can neck-, breast-or forehead flaps cover the defect, although they do not fulfill the demands for a satisfactory restoration of specific function. In the following work is the possibility of covering of a three layered facial defect with a combined skin muscle flap taken into consideration in view of the disadvantages of a simple or doubled skin flap. By this operation method the procedure consists of preparing various sized cervical island flaps which are connected with on the mastoid pedicled M. sternocleidomastoideus. We are describing a case of extensive tumorresection in the lip-chin area with neurofibromatosis Recklinghausen, whereby we have closed a large defect with two separately laid on muscle-pedicled, innervated island flaps from the fossa supraclavicularis. These bilateral compound flaps fulfill the demands of an immediate restoration of contour and prominence in this area. Apart from this they allow satisfactory voluntary activity, unpleasant dribbling of saliva is prevented.

[EEG-results in early-treated children with phenylketonuria 4 through 10 years under treatment (author's transl)].

EEG-results in 21 children with Phenylketonuria (PKU), put on a diet at least since their 3rd month of life, now aged 4 through 10 years, with normal psychomotor development, lacking abnormal neurological signs, are compared with the results in 796 healthy children of the same age. Visual evaluation and frequency analysis of the background activity of the EEGs reveal no differences between children with PKU and controls. Generally, there is no close correlation between mean plasma levels of phenylalanine (phe-means) during treatment, and the composition of the EEG when combined age groups are compared. But a trend can be demonstrated: In the small group of 8 years old children phe-means up to 6 mg-% can be associated with a faster (alpha), and phe-means above 6 mg-% with a slower (theta) background activity. The frequency of metabolic derailment (single phe-values above 10 mg-%) does not correlate with the EEG. Focal and generalized hypersynchronous activity (HSA) is observed significantly more often even in early treated, normally developing children with PKU. In children aged 4--8 years, HSA is associated with a high proportion of slow waves compared with those who do not display HSA. In children 9 and 10 years old, however, no such differences can be seen. It is hypothesised that even early treated children with PKU, in their first years show a retardation in the development of their background activity but catch up by 9--10 years. It still remains uncertain whether further age-appropiate development of the EEG can be observed after liberating or discontinuing the dietary regimen at this age.

Prolonged apnea in infant monkeys resulting from stimulation of superior laryngeal nerve.

We measured characteristics of apnea resulting from electrical stimulation of the superior laryngeal nerve (SLN) in 28 anesthetized infant monkeys ranging from a gestational age of 141 days to 49 days postterm and in four adult monkeys. Progressive reduction in ventilation accompanied weak suprathreshold SLN stimulation. Poststimulus apnea followed stronger stimulation. Apnea duration was directly proportional to stimulus duration, provided that stimulus intensity was at least 1.5 X threshold. Poststimulus apnea periods were briefer in older infants and did not occur in adults. Pao2 measurements in four animals showed that lower values were associated with the longer poststimulus apnea episodes. Upper airway resistance measurements obtained from a subglottal cannula revealed that glottal closure was transiently associated with the onset of SLN stimulation, but the degree of closure diminished during the course of stimulation and was not present during the subsequent period of apnea. The parameters of SLN stimulation determine specific changes in cardiovascular functions independently of their respiratory effects. The results suggest that a brief afferent input from the SLN may have prolonged effects on respiratory regulation in infants. These persisting effects may be an important consideration in sudden infant death syndrome mechanisms.

Synthesis of cold-insoluble globulin by cultured calf endothelial cells.

Radiolabeled amino acids were incorporated into nondialyzable protein by cultures of endothelial cells derived from calf aorta. Antibody prepared against purified bovine plasma cold-insoluble globulin (CIG) formed a strong precipitin line upon immunodiffusion against 3H-labeled proteins from endothelial cell culture media. This precipitin line formed a line of identity with the precipitin line formed by anti-CIG and purified CIG. Double-antibody immunoprecipitation of 3H-labeled protein showed that 36.3% of the radioactivity was specifically precipitated by the anti-CIG antibody. Gel filtration and polyacrylamide gel electrophoresis of the immunoprecipitate indicated that it contained a single protein species whose molecular weight was consistent with that of CIG isolated from plasma. When cultured cells were examined by indirect immunofluorescence microscopy, the endothelial cells stained specifically with anti-CIG antibody. These data indicate that cultured endothelial cells derived from calf aorta synthesize and secrete a protein antigenically similar to CIG.

Stimulation of fetal hemoglobin synthesis in baboons by hemolysis and hypoxia.

Fetal hemoglobin (Hb F) levels in the peripheral blood of baboons (Papio cynocephalus) increased from an average value of 0.78% to 18.1% during the recovery phase from phenylhydrazine-induced hemolytic anemia. A similar increase was observed in animals exposed to hypobaric hypoxia. Large individual variations in the maximal Hb F levels were observed which could not be correlated with the ages of the animals. Reinduction of hemolysis in two fully recovered animals resulted in Hb F levels that were of similar magnitude as in the preceding episode, suggesting the possibility of genetically determined individual variations in the rate of Hb F synthesis under the same conditions of erythropoietic stimulation. Reticulocytes from the animals subjected to hemolysis of hypobaric hypoxia synthesized similar absolute quantities of Hb F in vitro. The results of the present studies indicate that the physiological switch from the synthesis of Hb F to that of Hb A during ontogeny can be reversed in adult nonhuman primates by conditions of erythropoietic stress known to be associated with high erythropoietin levels. These findings open the possibility that Hb F synthesis in adult humans may be therapeutically modulated in individuals who might benefit from increased levels of Hb F, such as patients with sickle cell anemia.

Comparative effects of cocaine and pseudococaine on EEG activities, cardiorespiratory functions, and self-administration behavior in the rhesus monkey.

The effects of cocaine and pseudococaine on the EEGs, heart and respiratory rates, and self-administration behavior were studied in rhesus monkeys. An intravenous injection of cocaine (2.5 and 4.0 mg/kg) in the monkey produced low-voltage fast waves (LVFWs) in the EEGs and behavioral hyperexcitation accompanied by marked increases in the heart and respiratory rates with mydriasis and excessive salivation. In contrast, pseudococaine produced high-voltage slow waves (HVSWs) in the EEGs and behavioral depression accompanied by the same symptoms of the autonomic functions as those produced by cocaine. Both isomers were self-administered by the monkeys. During cocaine self-administration sessions, the animals showed hyperexcitation in their overall behavior, while with pseudococaine they showed almost normal behavioral responses. These results suggest that cocaine produced excitatory effects and pseudococaine inhibitory effects on the EEGs and behavior. Both isomers stimulate the heart and respiratory rates, and were self-administered by the monkeys.

[Lafora disease (author's transl)].

On the basis of 21 personal observations as well as those (82) from the litterature, it is concluded that the progressive myoclonic epilepsy with Lafora bodies (P.M.E.) constitutes a disease on its own. The clinical features are those described in the litterature observations and completed by some characteristics; the high frequency of visual symptoms (47 p. 100 personal cases); the relatively less bad evolution of epilepsy, perhaps in relation with use of modern drugs; the relatively moderate intensity of myoclonus which becomes complete only at the end of the evolution. From E.E.G. point of view, we can distinguish three periods: an initial one at the very onset of disease, who will show the same features as observated in primary generalized epilepsy, i.e. a well preserved background activity with superimposed generalized fast spikes and waves facilitated by the I.L.S. Then follows a period of evolutive E.E.G. (1-2 years after the onset of the disease) characterized by progressive slowing of the posterior background, enlargement of posterior slow activity and appearance of diffuse theta and delta activity. Simultaneously spikes and waves are taking less typical and bisynchronous aspect. Finally after 3 to 5 years from the onset there is a diffusely slow E.E.G. with superimposed fast multiple spikes. The E.E.G. findings in litterature usually refer only to this last period (stationary or terminal period). Occipital independent multiple spikes are frequently observed and could correlate with the visual symptoms observated in the Lafora disease. Some elements of differential diagnosis are given with respect to primary generalized epilepsy at the onset of the disease and later on with respect to dyssynergia cerebellaris myoclonica and to the progressive myoclonic epilepsy without Lafora bodies.

Immunochemical assays for an early diagnosis of endemic nephropathy.

Comparative studies aiming at the detection of certain tubular protein elements by means of Ouchterlony's immunodiffusion, in parallel with lysozyme and guanase assays, were carried out in the unconcentrated urines of 746 subjects, of whom 655 apparently healthy inhabitants (mostly children) from a region with endemic nephropathy (EN) and from Bucharest, as well as 91 adults with EN or various other diseases with renal involvement. The presence of light chains, of lysozymuria exceeding 2 microgram/ml, of beta2 microglobulin and of guanase in the urines of children and adults from the endemic area was significantly more frequent than in the control groups. These immunochemical changes are hence considered as valuable criteria for the detection of EN prior to the uremic stage. They should be looked for, first and foremost, in the young relatives of patients with this disease. In the stage of nitrogen retention the diagnostic value of these tests is reduced, since the same changes can also be found in the urines of patients suffering from other diseases with renal involvement, which show nitrogen retention as well.

[Dose distribution of photons and neutrons outside of the irradiation field of a 8-mev linear accelerator (author's transl)].

During the irradiation with a continuous radiation of 8 MeV by the linear accelerator LINAC SL 75/10, the dose distribution of photons and neutrons was measured outside of the irradiation field. The dose distribution of photons was mostly determined by thermoluminescence dose meters. There was a significant maximum of leak radiation on the back of the emitter head which showed a dose rate amounting to 7,5% of the dose rate of the central ray. Whereas the leak radiation on the patients' table was less than 0,1% of the dose of the central ray, the dose of scattered radiation emitted by the patient still amounts to 2% of the central ray dose in a distance of 15 cm from the field edge. The isodoses measured in the room demonstrate above all the importance of the lock for a sufficient radioprotection.--The dose rate of the neutrons formed in the shielding material by photonuclear reactions were measured mostly with uranium fission-track dosimeters. The highest equivalent dose rate was found on the back of the emitter head; it is about 17 rem/h. The maximum value of the equivalent dose rate of neutrons on the patient's table amounts to 1,4 rem/h and is almost independent of the size of the diaphragm. The isodoses in the room depend largely on the shape of the irradiation room; they confirm the existence of a sufficient radioprotection.

The organization, composition and matrix of hepatocyte nuclei exposed to alpha-amanitin.

Alterations in the structure and molecular composition of avian hepatocyte nuclei were compared following administration in vivo of lethal and sub-lethal doses of alpha-amanitin. This toxin interferes with extranucleolar transcription by direct inhibition of RNA polymerase II activity. the resultant effects include: extensive condensation of chromatin, displacement of nucleoplasmic contents and fragmentation of nucleoli. Changes in nuclear morphology were quantitated by stereometry and related to variations in RNA and residual, non-histone proteins (NHP). Gross alterations in nuclear structure and depletion of RNA and NHP levels were of similar magnitude with both doses of amanitin. The effects were fully reversible, however, with a minimal dose but terminal with a lethal dose. DNA and histone protein levels remained unchanged at all stages. These results imply that the process of transciption may itself keep and/or maintain chromatin in a dispersed state, and that in the absence of transcription chromatin naturally condenses. Modification of nuclear proteins may be necessary only to maintain chromatin compacted permanently or for extended periods of time. A model of nuclear organization is proposed to incorporate these considerations and to identify the probable location of the nuclear matrix in situ.

Studies on blood from the original Rhnull proposita and relatives.

A relative of the original Rh proposita, of group R1R2, shows weak expression of his Rh antigens, and is thought to be an Rhnull heterozygote. His wife and 3 of their 4 children show normal Rh antigen expression, but one daughter showed weak Rh antigen expression, as determined by quantitative haemagglutination. The observations support the proposition that the father is heterozygous for an unlinked modifier of Rh antigen expression. Stomatocytosis, observed in the Rhnull proposita and other Rhnull individuals, was also observed, but to a lesser degree, in the blood of an other individual thought to be an Rhnull heterozygote. This observation also supports the earlier conclusion that the Rhnull phenotype of the proposita is due to homozygosity for inactive alleles at a locus which controls the biosynthesis of precursor for Rh and LW antigens. Osmotic fragility tests showed that the Rhnull cells were more fragile than cells with normal Rh antigen expression, and cells from Rhnull heterozygotes had intermediate fragility. This is consistent with the proposition that Rh antigens are normal structural components of the red cell membrane, and the Rhnull heterozygotes show a deficiency of the Rh antigenic structures.

[Absorption and metabolism of glycerin in the neonatal period. I. Speed of turnover of glycerin during continuous intravenous infusions in newborn infants of various gestational ages and in older infants].

Glycerol was infused intravenously over 2 hours in preterm and term appropriate-for-date and in term small-for-date infants at the age of 12 to 72 hours and 10 to 14 days and in infants at the age of 3 to 8 months. The dosage was 0.25.kg-1.h-1 and 0.5.kg-1.h-1, respectively. Less than 6 per cent of the glycerol injused were recovered in the urine irrespective of the dosage. The total clearance was 9.1 to 14.6 ml.kg-1. min-1 during the first weeks of life with 0.25 g.kg-1.h-1 glycerol irrespective of gestational age and intra-uterine growth retardation; and it rose to 31.8 ml.kg-1.min-1 in older infants. With 0.5.kg-1.h-1 glycerol the total clearance values were lower in all groups. The glucose blood level and the blood lactate concentration as well as the parameters of the acid-base-balance were not significantly influenced by glycerol.

Structural alterations of the porcine heterograft after various durations of implantation.

Morphologic changes in six glutaraldehyde-fixed porcine heterografts recovered from five patients were studied with light and scanning electron microscopy. Light microscopy showed minimal changes in the valves recovered after 2 months of implantation. Fibrin deposits, red blood cell trapping, disruption of the valve matrix and interstitial edema were noted in heterografts 2 months and 2 years after implantation. In valves examined with scanning electron microscopy, 6 hours after implantation the surface changes varied from normal to patchy disruptions. Two months after inplantation, subendothelial fibers were conspicuous and the geometry of endothelial cells was changed. In valves examined after 6 months in place, many nuclei were burst and fibrin strands were seen over the surfaces. Bacterial infection was observed in two cases. The endothelial cells in the valve implanted for 2 years were completely obscured and some areas were densely covered with fibrillar material where blood cells were trapped in the mesh.

Urinary chromium excretion, diurnal changes, and relationship to creatinine excretion in healthy and sick individuals of different ages.

Since urine is the main excretory pathway for chromium, this study was conducted to compare in normal individuals the daily urinary chromium excretion with a 4 hr sample, to investigate diurnal fluctuations of urinary chromium and age-dependent relationship between urinary chromium and creatinine excretion. The results can be summarized as 1) there was no significant difference between the observed 24 hr chromium excretion and 24 hr excretion calculated from the one 4 hr samples, 2) a diurnal variation was observed when urinary excretion was expressed as chromium per minute, but no time-related variation could be established when chromium/creatinine (Cr/Cre) ratios in samples from three different periods of the same day were compared, although a significant positive correlation existed between urinary chromium and creatinine concentration, 3) the Cr/Cre ratio was found to be age-dependent, 4) in malnourished children the Cr/Cre ratio was very high and significantly different from that of normal infants, 5) This ratio for the eight diabetics was found to be significantly higher when compared with normal adults. On the basis of these results, it is suggested that morning 4 hr urinary chromium reflects the daily chromium excretion and that the Cr/Cre ratio of single urine samples obtained during this period is a reliable criterion in the evaluation of chromium nutrition of individuals in different conditions, provided that the influence of age is taken into consideration.

Tonic retinal influences in primates.

A type of unit discharge, termed "luxotonic," has been found in the striate cortex of unanesthetized squirrel monkeys and macaques.6, 21 The firing frequency of these units shows relatively little adaptation, continues indefinitely (hours), and reflects the level of diffuse illumination of the eye. The more numerous "photergic" units discharge more rapidly in the light, whereas "scotergic" units fire fastest in the dark (or at luminance levels below threshold for cones). Luxotonic activity is abolished by anesthesia and has not been described for striate cortex of other species. Primates also display a profound alteration in the EEG of striate cortex following elimination of all retinal input.32 Since this change is far more drastic than that produced by blindness in other species, it is natural to inquire whether it is related to the loss of the normally prominent luxotonic activity. When the blind monkey sleeps, the bizarre EEG is replaced by patterns wholly normal in appearance,32 indicating that some nonvisual system has extensive access to striate cortex in this state.

Relationship between gentamicin susceptiblity criteria and therapeutic serum levels for Pseudomonas aeruginosa in mouse infection model.

In this study estimations of in vivo and in vitro gentamicin susceptibility for a series of strains of Pseudomonas aeruginosa were compared. The series included an extremely susceptible strain, typically susceptible strains by current susceptibility criteria, and strains with enzymatic and permeability-mediated resistance. In vivo testing was done by using a mice protection test involving six 1-h doses of gentamicin and an inoculum of 50 50% lethal doses of P. aeruginosa. Both normal mice and cyclophosphamide-treated mice were used. It was found that peak serum levels and serum levels of gentamicin obtained just prior to the sixth dose (fifth dose trough levels) required for protection were much higher than minimal inhibitory concentrations (MICs) or minimal bactericidal concentrations (MBCs) obtained in high-cation medium. However, first dose trough levels were similar to MICs or MBCs. Only an extremely susceptible strain, 280, could be treated at antibiotic dosages and serum levels which are considered likely to be safe in humans. A distinct inoculum effect was found in the mice tests, with a 10-fold increase in inoculum producing a 4-fold increase in the amount of gentamicin required, but no inoculum effect was found for MICs. These results suggest that current susceptibility criteria in use for gentamicin and P. aeruginosa overestimate gentamicin susceptibility, particularly when low-cation growth medium is used for susceptibility testing and when treating disseminated infection.

[Nitrogen balance in adults and experimental study in rats fed with rice, beans and manioc flour supplemented with proteins].

Rice, beans and manioc flour are foods eaten daily in Northeast Brazil. Manioc flour is a source of energy, having a low protein content. There has been a great interest to supplement manioc flour with proteins. The present study was planned to show the nutritive value of a basic diet including rice, beans, manioc flour and other local foods, supplemented with different sources of proteins. The experiment was carried out in humans and rats. The supplements used were casein (Ca), isolated soy protein (PS) and a dry residue of a soymilk preparation (RSLS). Through nitrogen balance studies in adults it was shown that the diets with casein or isolated soy protein had a good absorption and retention of nitrogen. The addition of the soymilk residue resulted in a poor acceptance of the experimental diet; negative nitrogen retention was observed in the three persons who accepted the food and completed the balances studies. The same diets offered to humans, with the same protein supplements, were fed to rats. The nutritive value measured in the animals were similar, in all diets with all supplements, including the one with the dry residue of soymilk. The present results call the attention to the problems of transfering animal results to humans. It was also discussed the possibility of adding casein or isolated soy protein as an eventual protein supplement to manioc flour for human feeding.

[The myocardiopathies of hereditary neuro-muscular diseases].

This joint work has studied the cardiomyopathies occurring in hereditary neuro-muscular disorders (270 cases). The Duchenne type of disorder (74 cases) was responsible for asystole (4 cases), for cardiomegaly, and especially for abnormalities of the ECG (59 cases)--Q waves and large R waves in V1 and V6. The cardiomyopathy was of the hypokinetic type, with histological evidence of degeneration of the myocardial fibres. Dystrophia myotonica of Steinart (23 cases) caused conductive disorders (17 cases) which were either atrioventricular or intra-ventricular or both. Studies of the His pathway confirmed that these abnormalities were more diffuse in 5 cases. The main histological feature was interstitial fibrosis. There was a high risk of sudden death; ECG follow-up should be close. Friedreich's disease (20 cases) in its complete form led to later development of obstructive cardiomyopathy, with a systolic ejection murmur, cardiomegaly, and abnormalities of the ECG--left ventricular hypertrophy in the vertical axis, right ventricular and septal hypertrophy, repolarisation disorders similar to those found in coronary artery disease. Histology showed hypertrophy with degeneration of the myocardial fibres and interstitial fibrosis. This complete form was rare (7 cases out of 20); on the other hand, ECG abnormalites were very common (16 cases out of 20). The authors have tried to study the relationships between primary cardiomyopathies (50 cases) and peripheral neuromuscular disorders. 17 of the 39 peripheral muscle biopsies were abnormal, but a well-defined muscular dystrophy could not be found in them.

[The exercise test in congenital aortic stenosis].

Exercise tests can be carried out without risk in a child or young adult with asymptomatic congenital aortic stenosis. In such a case, the ECG at rest gives only a very poor indication of the severity of the stenosis, which often becomes worse with age. The only sinister findings is that of repolarisation defects, which are almost always absent at rest. Exercise, by causing an imbalance between myocardial oxygen consumption and oxygen transport, will cause "ischaemic" signs to appear on the ECG. At the same time, the absence of compensation of pressure is a very important sign of poor tolerance; the test can usefully be concluded by using a floating catheter to measure the pulmonary capillary pressure. The correlation between a positive test and the stenotic pressure gradient, measured on catheterisation at rest, is good; but "negative" tests may be found in certain cases with a tight stenosis, especially if maximal rate could not be reached. In cases of stenoses which were initially mild or moderate, this test is very useful as a follow-up procedure, as such stenoses may become worse with age and require surgical correction. It is also useful for assessing the results of surgery, and of rehabilitation.

Prostaglandin E2 biosynthesis and effect in pigeon aorta. Possible role in atherogenesis.

This study for the first time examines the biosynthesis and effect of prostaglandin E2 (PGE2) in aorta during genetic atherosclerosis. Biosynthesis of PGE2 from [1-14C]arachidonic acid was investigated in the aorta of spontaneously atherosclerosis-susceptible White Carneau pigeons and was compared with that of the atherosclerosis-resistant Show Racer breed. Most of the PGE2 synthetase activity was located in the microsomes. The synthesis was linear up to an hour and was optimal at pH 7.4. The formation of PGE2 in the aorta in the White Carneau pigeons was significantly higher (P less than 0.01) than that in age-matched Show Racer pigeons. In vitro PGE2 strongly inhibited the cholesteryl ester hydrolase activity (51.6% inhibition at 4 X 10(-7) M concentration) in the supernatant fraction of the aorta. On the basis of (1) the increased formation of PGE2 in the aorta of atherosclerosis-susceptible pigeons and (2) its effect on specific enzymes that control cholesteryl ester concentration in aorta, it is hypothesized that PGE2 synthesized at a higher rate in damaged aorta has a significant role in cholesteryl ester accumulation during atherogenesis.

Bacteriochlorophyll fluorescence of purple bacteria at low redox potentials. The relationship between reaction center triplet yield and the emission yield.

This work describes fluorescence yield measurements in suspensions of strains of Rhodospirillum rubrum and Rhodopseudomonas sphaeroides in which the iron . quinone complex (X) was chemically reduced (state [PIX-]; P is the reaction center bacteriochlorophyll dimer, I is the long wavelength bacteriopheophytin), and compares these with the fluorescence observed when all the traps are open (state [PIX]) and with the fluorescence observed when all the traps are closed (state [P+IX]). At 77 K the amplitude and the shape of the fluorescence emission spectrum in [PIX-] are identical to those observed in [PIX]. This is a strong indication that all the extra fluorescence observed at room temperature in [PIX-] is, in fact, caused by an efficient back reaction [P+I-X-] leads to [P*IX-]. Using an equation similar to the original Vredenberg-Duysens relationship (Vredenburg, W.J. and Duysens, L.N.M. (1963) Nature 197, 355-357) but now assuming that a single reaction center has a probability pt of trapping an excitation and (1--pt) of re-emitting it to the surroundings, we are able to calculate pt as a function of the temperature by measuring the fluorescence in [PIX], [PIX-] and [P+IX] as a function of the temperature. The calculated pt values agree reasonably well with triplet yields measured in isolated reaction centers. Finally, we have measured the reaction center triplet yield (PTR) in intact systems and we have shown that the sum of the triplet yield and the remaining loss processes (PL) in the antenna bacteriochlorophyll including the bacteriochlorophyll dimer (such as fluorescence, internal conversion or direct triplet formation) is approximately constant; if we assume that at 77 K the only process which occurs in the reaction center is the formation of a reaction center triplet, than PTR + PL=1. The energy barrier between [P*IX-] and [P+I-X-] was estimated to be 0.11--0.15 eV for a set of preparations.

Phycobiliprotein synthesis in protoplasts of the unicellular cyanophyte, Anacystis nidulans.

Stable and metabolically active protoplasts were prepared from the unicellular cyanophyte, Anacystis nidulans, by enzymatic digestion of the cell wall with 0.1% lysozyme. The yield of protoplasts from intact algal cells was approx. 50%. Incorporation of L-[U-14C]leucine into cold trichloroacetic acid-insoluble material from protoplasts preparations was linear for 1.5 h and continued for an additional 2.5 h. Incorporation of radiolabeled leucine into hot trichloroacetic acid-insoluble material from protoplast preparations demonstrated protein synthesis in protoplasts in vitro. Phycocyanin is the principal phycobiliprotein and allophycocyanin is a minor phycobiliprotein in A. nidulans cells. The light-absorbing chromophore of both of these phycobiliproteins is the linear tetrapyrrole (bile pigment), phycocyanobilin. Radiolabeled phycocyanin and allophycocyanin were isolated from protoplast preparations which had been incubated with L-[U-14]leucine or delta-amino[4-14C] levulinic acid (a precursor of phycocyanobilin). The radio-labeled phycobiliproteins were purified by ammonium sulfate fractionation and ion-exchange chromatography on brushite columns. The specific radioactivity of phycocyanin and allophycocyanin in brushite column eluates (protoplasts incubated with radiolabeled leucine) was 106 000 and 82 000 dpm/mg, respectively. The specific radioactivity of phycocyanin and allophycocyanin in brushite column eluates (protoplasts incubated with radiolabeled delta-aminolevulinic acid) was 33 000 and 38 000 dpm/mg, respectively. Phycobiliproteins from protoplasts incubated with radiolabeled leucine were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 25% of the incorporated radioactivity in protoplast lysates and approx. 60% of the incorporated radioactivity in protoplast lysates and approx. 60% of the incorporated ratioactivity in phycocyanin and allophycocyanin (in brushite column eluates) comigrated with the subunits of these phycobiliproteins on sodium dodecyl sulfate-polyacrylamide gels. Chromic acid degradation of phycobiliproteins from protoplast preparations incubated with delta-amino[4-14C] levulinic acid yielded radiolabeled imides which were derived from the phycocyanobilin chromophore. Imides from radiolabeled phycobiliproteins isolated from protoplast preparations incubated with L-[U-14C]leucine did not contain radioactivity. These results show that both the apoprotein and tetrapyrrolic moieties of phycocyanin and allophycocyanin were synthesized in A. nidulans protoplasts in vitro.

Vitamin A and thyroxine carrier proteins in chicken plasma: steady-state control of the plasma level of free retinol-binding protein and free thyroxine.

1. The binding parameters of prealbumin-2 with retinol-binding protein and thyroxine (T4) revealed the existence of distinct and multiple sites for both retinol-binding protein and T4. 2. From the analysis of binding parameters of retinol-binding protein with prealbumin-2 it is clear that under steady-state conditions about 99% of the holo-retinol-binding protein remains bound to prealbumin-2. 3. Equilibrium dialysis studies on binding properties of thyroid hormones with prealbumin-2 revealed that it has a single high affinity site and three low affinity sites. 4. The occurrence of three carrier proteins for thyroid hormones, thyroxine-binding globulin, prealbumin-2 and albumin has been demonstrated. However, the chicken thyroxine-binding globulin differs from human thyroxine-binding globulin by being relatively less acidic and occurring at a two-fold lower concentration. But the thyroid hormone binding parameters are comparable. 5. Highly sensitive methods were developed for determination of T4 binding capacities of the various proteins and plasma level of total T4 by fractionation of carrier proteins and further quantitatively employing in electrophoresis and equilibrium dialysis. 6. The thyroxine-binding proteins were found to be of two types, one (viz., thyroxine-binding globulin) of great affinity but of low binding capacity, which mainly acts as reservoir of T4, and another (viz., prealbumin-2) of low affinity but of high binding capacity, which can participate predominantly in the control of the free T4 pool.

Nitrogen fixation and hydrogen metabolism in photosynthetic bacteria.

The photosynthetic bacteria are found in a wide range of specialized aquatic environments. These bacteria represent important members of the microbial community since they are capable of carrying out two of the most important processes on earth, namely, photosynthesis and nitrogen fixation, at the expense of solar energy. Since the discovery that these bacteria could fix atmospheric nitrogen, there has been an intensification of studies relating to both the biochemistry and physiology of this process. The practical importance of this field is emphasized by a consideration of the tremendous energy input required for the production of artificial nitrogenous fertilizer. The present communication aims to briefly review the current state of knowledge relating to certain aspects of nitrogen fixation by the photosynthetic bacteria. The topics that will be discussed include a general survey of the nitrogenase system in the various photosynthetic bacteria, the regulation of both nitrogenase biosynthesis and activity, recent advances in the genetics of the nitrogen fixing system, and the hydrogen cycle in these bacteria. In addition, a brief discussion of some of some of the possible practical applications provided by the photosynthetic bacteria will be presented.

Stimulation of acetylcholine output from brain slices caused by the ionophores BrX-537A and A23187.

1 The effect of two ionophores, BrX-537A (Bromolasolacid) and A 23187, on acetylcholine (ACh) output from brain slices was studied. 2 The slices were prepared from rat cerebral cortex, incubated in Krebs solution containing physostigmine and ACh output determined by bioassay. 3 Both ionophores enhanced ACh output. BrX-537A exerted its maximal effect, a six fold increase, at a concentration of 1.8 micron, while A 23187 caused a three fold increase at a concentration of 58 micron. 4 When the slices were incubated in a Ca-free medium, the effect of A 23187 on ACh output was only reduced, BrX-537A was abolished while that of BrX-537A was also active when disodium edetate (EDTA) was added to the the Ca-free medium. 5 The activity of BrX-537A was not affected by the presence of tetrodotoxin in the incubation medium. 6 The stimulation of ACh output elicited by KCl (25 mM) was increased further by hyoscine, but not by BrX-537A. Hyoscine however had no effect when ACh output was stimulated by BrX-537A. 7 The effect of BrX-537A on ACh output was potentiated by the addition of Mg2+ (9.3 mM) to the incubation medium and was reduced in a Mg-free medium. 8 It is concluded that A 23187 stimulates ACh output by transporting extracellular Ca2+ into cholinergic nerve endings. The effect of BrX-537A does not depend only on Ca2+ but also on other mechanisms.

Biological nitrogen fixation in the terrestrial environment of a high Arctic ecosystem (Truelove Lowland, Devon Island, N.W.T.).

Arranged in descending order of nitrogen-fixing (acetylene-reducing) potential the sites examined were mesic meadow and peat polygon troughs (equal rank), transition zone between mesic meadow and gravel ridge, gravel ridge, polar dessert, and peat polygon tops. The dominant nitrogen-fixing microorganisms, as in other Arctic areas, were blue-green bacteria, especially those epiphytic on Arctic mosses. The epiphytic association exhibited an optimum temperature for fixation of 20 degrees C. Other bacteria potentially able to fix nitrogen were present in the soils examined but their activity was severely restricted by low soil temperatures and lack of readily utilizable energy sources. These bacteria included members of the genera Klebsiella (the most numerous), Bacillus, Clostridium, and Beijerinckia (scarce). Also present at many of the sites was an unidentified yellow-pigmented fixer which was not Mycobacterium flavum. All fixers were psychotrophic rather than psychrophilic, having an optimum temperature greater than 20 degrees C but capable of slow growth at 5 degrees C or lower. The rate of acetylene reduction by the epiphytic system increased with the number of successive exposures to acetylene, a phenomenon of some significance in any calculations designed to measure the amount of nitrogen fixed in certain ecosystems.

Single-radial-complement-fixation: a new immunodiffusion technique. Assay of the antibody response to the type-specific antigens of influenza virus in post-infection human sera.

A new immunodiffusion technique in agarose gel for the quantification of complement-fixing antibodies is described. The test involves the incorporation of antigen and complement in a primary agarose gel. Heat-inactivated serum samples are allowed to diffuse radially from wells overnight at 4 degrees C. A secondary gel, containing antibody-coated sheep erythrocytes, is layered on top of the first gel and the system is incubated for 45 min at 37 degrees C. Where complement is fixed, i.e., around wells with positive serum samples, zones of unlysed cells appear. There is a straight line relationship between zone areas so produced and log(2) serum titres obtained with the conventional complement fixation test. The method appears to be applicable to a variety of antigens. It has been found suitable for bacterial and viral antigens. The test can also be reversed, thus allowing the quantification of diffusible antigens in a gel containing immune serum and complement. This paper describes in detail the use of this method as a diagnostic tool for the assay of complement-fixing antibodies to the type-specific antigens of influenza virus in paired human sera.

The immunogloblin nature of nephritic factor (NeF).

NeF was shown to be antigenically and structurally similar to IgG by the following experiments: (1) NeF activity in serum was absorbed by and, under acid conditions, could be eluted from (a) anti-myeloma IgG antibody coupled to Sepharose and (b) protein A-Sepharose. (2) Purified NeF could bind to anit-myeloma IgG-Sepharose and could be eluted with acid, and this binding was blocked by myeloma IgG. (3) An antibody to beta2, microglobulin, showing strong cross-reactivity with normal IgG, bound NeF activity before, but not after, absorption of the antiserum with IgG. (4) Sepharose-coupled antibodies to NeF could bind activity which was recovered in the acid eluate. This binding capacity was lost after absorption of the antibody with normal and myeloma IgG. (5) Structural similarity was demonstrated by pepsin and papain digestion, which resulted in NeF activity eluting with F(ab')2 and Fab fragments from protein A-Sepharose and Sephadex G-150. (6) Autoradiography of PAGE-SDS of 125I-labelled NeF eluted from EA43bBb cells showed that NeF had a larger H chain than normal IgG, suggesting that NeF might be an abnormal IgG molecule.

[The significance of rhythmic mid-temporal discharges (author's transl)].

Out of 50,000 EEG's those of 38 subjects contained rhythmic mid-temporal discharges (RMTD), corresponding to an incidence of 0,1%. The morphological features of RMTD are: 1. frequency: 5.5-6.5/sec 2. shape: monophasic and regular with occasionally interposed 12/sec. activity. 3. localisation: mid temporal, often spread to anterior, seldom posterior region. 4. occurence: bilateral, simultaneous, or alternating sides. They are closely linked to the drowsy state, occuring at the transition from A2 to B2 stage (IA2) and arising from a fairly desynchronized EEG. RMTD are commonly seen within REM periods, which are markedly fragmented with interspersed periods of drowsy patterns lasting 20-90 sec, during which the RMTD are seen. Occasionally they are strictly related to slow eye movements and periodic respiration. The RMTD are an individual feature, appearing in different persons with variing penetrance. Their occurence is favoured or inhibited to a certain degree by external circumstances. Slow and fast wave sleep in subjects with RMTD are disturbed. Both of them, especially the fast wave sleep are reduced in favour of markedly increased stages of drowsiness with RMTD, which sometimes last several minutes. In spite of such abnormal organisation of sleep the subjects feel recovered in the morning and sleep disturbances are not reported. RMTD could therefore be considered as "bioelectrical sleep disorder". We did not find any correlation between RMTD and clinical findings, in particular not with psychomotor or any other form of epilepsy.