An introduction to a series of essays honoring Erich Meyerhoff (1919–2015), AHIP, FMLA, who was active in and contributed to the Medical Library Association for generations. Journal of the Medical Library Association (JMLA), to catch a glimpse of him and understand why he is honored with a festschrift sponsored by the Fellows of the Medical Library Association (MLA).A few times in each generation, we count ourselves fortunate if we have had the opportunity to know an extraordinary individual. Such an individual was Erich Meyerhoff, AHIP, FMLA (1919–2015), who marked his time not simply by living a very long life, but by doing, creating, enabling, and inspiring those around him. He had a passion for his profession and the individuals who made up that profession. Those of us who were not fortunate to know him personally now have the opportunity, through the series of essays published in this issue of the JMLA, provided an overview of his career and, most notably, comments from his peers [Erich was active in and contributed to MLA for generations. His legacy is one of unparalleled support for and encouragement of MLA and its members. His obituary, published in is peers . DescripThe series of essays in this festschrift provides a snapshot of an extraordinary individual who exemplified professionalism and an abiding faith in social justice and the possibilities of what an organization such as MLA could achieve in the twentieth and twenty-first centuries. This festschrift is meant not only to honor Erich, but to provide an example of a professional life well lived for current and future members of MLA. His activities and accomplishments were recognized by MLA through the Marcia C. Noyes Award, the Janet Doe Lectureship, and MLA Fellowship, but there was much more to Erich than these important awards or his longevity.In their essay, “Erich Meyerhoff: A Man for All Medical Librarians,” Judith Messerle, AHIP, FMLA, and Lucretia W. McClure, AHIP, FMLA, provide a sketch of his life, education, career, and accomplishments, which spanned many significant developments in health sciences librarianship . They noUnion Catalog of Medical Periodicals and lasted as a resource center and service bureau from 1960 to 2003.Patricia E. Gallagher, AHIP, FMLA, traces the development of the Medical Library Center of New York in her essay, “Library Resource Sharing and the Medical Library Center of New York” . The cenElaine Russo Martin, FMLA, defines and reviews democratic librarianship in her essay, “Democratic Librarianship: The Role of the Medical Library in Promoting Democracy and Social Justice” . ThroughWayne J. Peay, FMLA, and Helen-Ann Brown Epstein, AHIP, FMLA, update the now historic tenth Janet Doe Lecture, which Erich presented in 1977, in their essay, “The Tenth Doe Lecture: A Forty-Year Perspective: Still Relevant after All These Years” . The lecFinally, Stephen J. Greenberg, AHIP, provides a window into the world of historical writing and how it changed over the course of Erich’s life in his essay, “Medical History: As It Was; As It Will Be” . GreenbeThis series of essays honoring Erich Meyerhoff grew out of discussions of the MLA Fellows group during their annual meeting in conjunction with the MLA annual meeting. A small task force was established to define topics for the festschrift essays and engage authors. I thank the members of the Meyerhoff Task Force: Helen-Ann Brown Epstein, AHIP, FMLA, Patricia E. Gallagher, AHIP, FMLA, Stephen J. Greenberg, AHIP, J. Michael Homan, AHIP, FMLA (chair), and Lucretia W. McClure, AHIP, FMLA.J. Michael Homan, AHIP, FMLA,homan@mayo.edu, Emeritus Director of Libraries, Assistant Professor of Biomedical Informatics, and Emeritus Consultant, Health Sciences Research, Mayo Clinic, Rochester, MN


Pundamilia pundamilia and P. sp. ‘red head’). The SNP markers were distributed on 22 linkage groups and the total map size was 1,594 cM with an average marker distance of 1.01 cM. This high-resolution genetic linkage map was used to anchor the scaffolds of the Pundamilia genome and estimate recombination rates along the genome. Via QTL mapping we identified a major QTL for sex in a ∼1.9 Mb region on Pun-LG10, which is homologous to Oreochromis niloticus LG 23 (Ore-LG23) and includes a well-known vertebrate sex-determination gene (amh).Genetic linkage maps are essential for comparative genomics, high quality genome sequence assembly and fine scale quantitative trait locus (QTL) mapping. In the present study we identified and genotyped markers via restriction-site associated DNA (RAD) sequencing and constructed a genetic linkage map based on 1,597 SNP markers of an interspecific F2 cross of two closely related Lake Victoria cichlids ( Paralabidochromis chilotes and Paralabidochromis sauvagei and contained 184 microsatellites and two SNP markers with a mean marker spacing of 6.09 cM on 25 linkage groups , namely LG 1 (XY), LG 3 (ZW), and LG 23 (XY)  and the anti Müllerian hormone (amh) might have been re-used as sex determination loci . The map was built using 1,597 SNPs identified and genotyped via restriction-site associated DNA (RAD) sequencing with an average marker distance of 1.01 cM. We then used the linkage map to anchor the scaffolds of the P. nyererei reference genome to the 22 linkage groups of the map and to perform a QTL analysis for putative sex determination loci in Pundamilia. We identify the LG determining sex in a Lake Victoria cichlid cross, as well as potential candidate genes for sex determination and put these findings into the context of sex determination evolution within a rapidly radiating clade of fish.In the present study we construct a linkage map of an interspecific F2 cross between two very closely related Lake Victoria cichlid species (Pundamilia sp. ‘red head’ (th or 5th lab generation) and a wild P. pundamilia female caught by OS at Makobe Island in Lake Victoria in 2003. Eggs were removed from the female’s mouth five days after spawning and reared in isolation from the adults. After reaching maturity, four F1 individuals were crossed, resulting in two F2 families with together more than 300 individuals. When F2 individuals were adult and sexually mature sex was determined based on coloration, then sedated fish were killed with MS222 , and a fin clip was taken and stored in 98% ethanol for genetic analyses . Genomic DNA of 218 F2 progeny, the four F1 parents, and the two F0 grandparents was extracted using phenol-chloroform  each on a single lane of an Illumina HighSequation 2500 platform either at the Next Generation Sequencing Platform of the University of Bern or at the Genomic Technologies Facility of the University of Lausanne. Some individuals and all F0 grandparents were sequenced in up to three libraries to increase coverage. Together with each library, we sequenced about 10% reads of bacteriophage PhiX genomic DNA (Illumina Inc.) to increase complexity at the first 10 sequenced base pairs. During read processing, PhiX reads were further utilized to recalibrate libraries to equalize base quality scores across Illumina lanes utilizing GATK version 3.2  and filtered using FASTX Toolkit 0.0.13 (http://hannonlab.cshl.edu/fastx_toolkit/index.html) requiring a minimum quality of 10 at all bases and of 30 in at least 95% of the read. After PhiX removal, reads were demultiplexed, cleaned, and trimmed to 92 bp with process_radtags implemented in Stacks v1.26 (P. nyererei reference genome (https://github.com/jpuritz/dDocent/blob/master/scripts/dDocent_filters), genotypes were further excluded (thresholds given in brackets) on criteria related to allelic balance at heterozygote sites , quality vs. depth (ratio <0.5), strand presentation (overlapping forward and reverse reads), and site depth . Multi-allelic variants and indels were removed, resulting in 7,401 SNPs. Of the 7,401 filtered SNPs 2,052 were alternative homozygous in the grandparents, and were used to build the genetic linkage map.Before recalibration, read qualities were inspected using fastQC (e genome . Mapped P < 0.001) were excluded for linkage map reconstruction. Linkage groups were identified based on an independent logarithm of odds (LOD) threshold of 12. Unlinked markers were excluded. The strongest cross-link (SCL) in the final map is 5.4. The linkage map was built using the regression mapping algorithm, a recombination frequency smaller than 0.40, and an LOD larger than 3. Up to three rounds of marker positioning were conducted with a jump threshold of 5. A ripple was performed after the addition of each new marker. Map distances were calculated using the Kosambi mapping function. All markers resolved onto 22 linkage groups were matched to positions in the Oreochromis niloticus genome using a chain file  identified during mapping (see paragraph above). We ordered and oriented the scaffolds with ALLMAPS (Pundamilia nyererei reference (Pundamilia reference version 2.0). Chain files were produced with ALLMAPS and in the opposite direction using chainSwap from kentUtils (https://github.com/ENCODE-DCC/kentUtils). We could then use the chain file to liftover the position of all 7,401 genotyped loci, using Picard liftoverVcf (http://broadinstitute.github.io/picard/index.html). In addition, we generated a new version of the NCBI Pundamilia nyererei RefSeq annotation file with the positions for reference version 2.0 by lifting over the positions from the NCBI PunNye1.0 annotation release 101 (https://www.ncbi.nlm.nih.gov/genome/annotation_euk/Pundamilia_nyererei/101/#BuildInfo) using the UCSC liftOver tool  are deposited on short read archive SRA accession SRP136207. Supplemental material available at Figshare: https://doi.org/10.25387/g3.6221921.All genomic resources see , the genPundamilia linkage map, 1,182 markers could be positioned onto Oreochromis niloticus linkage groups (Ore-LG). The linkage map comprises 22 linkage groups containing 1,597 markers with an average marker distance of 1.01 cM adding up to a total map length of 1593.72 cM , Table 1Pundamilia linkage map provides a new chromosome framework for whole genome sequence assembly and map integration with more anchoring points then previous published maps. The anchored genome encompasses 78.7% of the total bases  of the original P. nyererei reference genome based on 383 anchored scaffolds, of which 233 are now oriented. This is a slightly higher fraction than in the Lake Malawi cichlid Metriaclima zebra, where 564,259,264 bp (66.5%) of the genome sequence could be anchored to linkage groups . We did not find any further associations on any of the other LGs. Ore-LG23 has been previously identified as one potential XY sex-determining LG in Oreochromis . This suggests that none of the markers used to build the linkage map are determining sex directly, but that the causal locus can be found close by and indicates that there are no further major genetic determiners of sex segregating in this cross. Investigating the segregation patterns in the larger of the F2 mapping-families (n = 122) more in detail revealed that the loci selected to build the map  and male (BB) and heterozygous in both F1 (AB)) segregate as expected in a 50:50 ratio of AA:AB in F2 females and AB:BB in F2 males . Among them is the anti-müllerian hormone (amh), a master gene for sex determination in other fish. Amh is part of the transforming growth factor beta pathway, responsible for the regression of Müllerian ducts in tetrapods , a mutation in the receptor of the amh (amhrII) determines sex (amhy (Y chromosome-specific anti-müllerian hormone) gene has been inserted upstream of amh in the cascade of male development in the neotropical silverside Odonthestes hatcheri  and in Astatotilapia calliptera on LG7 (gsdf) (Pundamilia Pun-LG10 (Ore-LG 23) acts as an (evolving) sex chromosome, even though it might not be the only region controlling sex in Pundamilia. The anti-müllerian hormone amh (or a derived copy) appears to be a strong candidate influencing sexual development in Pundamilia, but further work is warranted to characterize the genomic candidate region and the impact of this candidate gene on sex determination.Within our mapping interval of ∼1.9 Mb, 65 genes, based on the NCBI annotation for the new 7 (gsdf) . Our resamh, nor Pun-LG10 or a homologous region was invoked in sex determination in other Lake Victoria cichlids that have previously been used for mapping sex (A recent meta-analysis showed that transitions between sex determination systems are frequent across fish species, including transitions to and between heteromorphic sex chromosomes (ping sex , this ma


In this paper, an application for the management, supervision and failure forecast of a ship’s energy storage system is developed through a National Marine Electronics Association (NMEA) 2000 smart sensor network. Here, the NMEA 2000 network sensor devices for the measurement and supervision of the parameters inherent to energy storage and energy supply are reviewed. The importance of energy storage systems in ships, the causes and models of battery aging, types of failures, and predictive diagnosis techniques for valve-regulated lead-acid (VRLA) batteries used for assisted and safe navigation are discussed. In ships, battery banks are installed in chambers that normally do not have temperature regulation and therefore are significantly conditioned by the outside temperature. A specific method based on the analysis of the time-series data of random and seasonal factors is proposed for the comparative trend analyses of both the battery internal temperature and the battery installation chamber temperature. The objective is to apply predictive fault diagnosis to detect any undesirable increase in battery temperature using prior indicators of heat dissipation process failure—to avoid the development of the most frequent and dangerous failure modes of VRLA batteries such as dry out and thermal runaway. It is concluded that these failure modes can be conveniently diagnosed by easily recognized patterns, obtained by performing comparative trend analyses to the variables measured onboard by NMEA sensors. Vessels are essentially floating complex systems, such as freighter ships, carrier ships, cruise ships, ship factories, oceanographic research ships, and battle ships, etc. All of these vessels are equipped with navigation, propulsion, power generation, distribution, and other systems of life support. For the control and supervision of such systems, the existence of common standards of communication networks would be desirable. As in other distributed control schemes, the transfer of data in ships is done with networks of low-level control, of real-time features, and of critical natures, with temporary response restrictions. They include less restrictive hierarchical networks not directly related to the safety of the ship, up to supervision, planning, and business management systems.Decision-making that affects the operation of the ship, involving safety, crew, passengers, cargo and the environment, is generally performed on the bridge or other control centers. To make decisions efficiently and carry them out requires access to many of the information systems on board. Systems of special relevance can include navigation, weather forecasting, power generation and storage, engines and machinery, data processing, messages and alarms, etc. The special risks involved in an aggressive environment such as marine navigation require adequate levels of security, availability, redundancy, and latency of communications networks. Although local office-type networks can work at the same speed, from the point of view of reliability they are unadvisable.A large number of sensors, controllers, electronic devices and systems must be installed on shipboard, and they generate a huge amount of data and information. Such information must be exchanged reliably to aid system integration and safe navigation ,2. TheseSpecifically, issues concerning power generation, status supervision, alarms, prognosis, and the control of devices such as generators, alternators, hybrid inverters, and shore-power, are tasks to be considered. In addition, electrical distribution tasks such as the delivery of power on vessels, identification of loads, load sharing, and virtual breakers are important issues.Ship navigation involves different critical operations where no uncertainty in decision making is acceptable. Such uncertainty can subject crew, passengers, cargo, or the ship itself to conditions of potential danger. A diagnosis of predictive failures is essential for a safe navigation. This diagnosis is based on the permanent analysis of the health condition of the system, through the detection and analysis of different symptoms that precede system’s failures. This analysis can be done using time-series data of certain system parameters. These techniques for predictive diagnosis allow for generally adequate margins or reaction capacities in the application of predictive maintenance operations.In order to maintain a safe navigation, ships require that many systems are continuously available, i.e., navigation, communication, and weather forecast equipment. If a failure of the storage and power supply system affects the maneuvering system during a critical operation, the result can be a serious accident . TherefoIn ships, battery banks are installed in chambers that normally do not have temperature regulation and that are therefore significantly conditioned by the outside temperature. In this paper, a specific method based on the analysis of the time-series data of random and seasonal factors is proposed for the comparative trend analyses of the internal temperature of batteries and the battery installation chamber temperature. The objective is to apply a predictive fault diagnosis to detect any undesirable increase in battery temperature due to a prior heat dissipation process failure, to avoid the development of the most frequent and dangerous failure modes of VRLA batteries—such as dry out and thermal runaway. It is concluded that these failure modes can be conveniently diagnosed by easily recognized patterns obtained by performing comparative trend analyses to the variables measured onboard by National Marine Electronics Association (NMEA) sensors.The advantage of the diagnostic method based on the comparative trend analyses proposed in this work is that it focuses on the assignable cause that precedes the dry out and thermal runaway failures, which is the loss of the operating capacity of the heat dissipation process of the battery banks. The heat dissipation process of battery banks in real navigation conditions does not have a specific nature. It depends on the characteristics of the installation chambers (which do not remain invariable in time), the materials used, the size of the heat dissipation areas, and the ventilation conditions, etc. All these factors affect the batteries in a variable way, depending on the type of batteries and their intrinsic aging conditions.In this study the operation and tests of the batteries have been made in real navigation conditions using a NMEA 2000 sensor network. As stated before, the possibility of using refrigerated chambers for battery installation is not feasible in most cases, nor from the economic point of view or from the energy consumption point of view.In Before the year 2000, some attempts of standardizing communication networks in ships were made. Various manufacturers developed different systems, however, differences in requirements made it difficult to find one common standard for the networks used in the different parts of the ship. IEC 61162-1 defines about 50 talker identifiers for individual types of navigation and radio communication equipment alone. In  approximWith regard to maritime navigation, a new standard for linear network communication—NMEA 2000—was presented in 2001 by the National Marine Electronics Association (NMEA) ,6. It waSome features of the data connection layer of the NMEA 2000, like in the physical layer, are determined by the choice of CAN as the main network. The NMEA 2000 fully makes use of the international standard of the ISO 11783-3 data connection layer, which is virtually identical to the SAE J1939-21 standard . Additional requirements contained in the NMEA 2000 ensured better copying, with special types of data and formats transmitted through a navigational device and supported by the special construction of such devices [In 2007 CANopen as IEEE P1551.6 was proposed as belonging to the IEEE 1451 family of Smart Transducer Interface Standards for sensors and actuators ,11. Part 1: IEC 61162-1 single talker and multiple listeners .Part 2: IEC 61162-2 single talker and multiple listeners, high-speed transmission.Part 3: IEC 61162-3 serial data instrument network .Part 450: IEC 61162-450 multiple talkers and multiple listeners–Ethernet interconnection .Concurrently, taking in mind mainly big ships, the Working Group 6 (WG6) of Technical Committee 80 (TC80) of the IEC defined a set of IEC 61162 standards for “Digital interfaces for navigational equipment within a ship”, divided into four parts :Part 1: Subgroup TC80/WG6 specified the use of Ethernet for on-board navigation networks. The specification was limited to the transport of NMEA sentences subject to the definition made in 61162-1 on IPv4. Due to the low amount of complexity of the protocol, it was called Lightweight Ethernet (LWE), and was presented at the ISIS 2011 symposium .IEC 61162-460: 2015 (E) is a complement to the IEC 61162-450 standard, seeking the introduction of higher security standards, and improving network integrity. The first edition was published in 08/2015.Some of the first ship data network standards published were the US Navy’s SAFENET (Survivable Adaptable Fiber optic Embedded Network) standards I and II  in 1988,In the period 1991–1993 a Norwegian research project, MITS (Maritime Information Technology Standard) was developed and subsequently, between 1993–1996 the MITS protocol was implemented on several ships, based on a single non-redundant Ethernet on the physical layer and TCP/IP protocols up to the transport layer ; howeverStandard redundancy support and fully redundant network system based on dual Ethernets and the IP protocols were specified in the project PISCIS (1998–2000), taken up and developed by the IEC 61162-400 series of standards .The new work item 80/506/NP on an Ethernet based interface standard was proposed in 2007 by Sweden to the IEC and accepted in March 2008. The IEC TC80/WG6 went to work on its development and the final version was published in April 2011 as IEC 61162-450 . This TC80/WG6 subgroup has specified with IEC 61162-450 and IEC 61162-460 standards the use of Ethernet and safety security conditions for shipboard navigational networks, taking into account the new developments in legislation that make it necessary to look closer at improved system integration tools such as data networks ,19.Other contributions in the context of IT standards were made through major integration projects such as Flagship, especially in the development of IEC 61162-450 and the final stages of ISO 28005 [Currently, there are projects such as Signal K, which has been proposed as a solution of the next generation for the exchange of marine data. It not only allows communication between instruments and sensors aboard a single ship, but also aims to share data among several ships, navigation aids, bridges, marinas and other land resources. It is designed to be easily used by web and mobile applications and to connect modern ships to the concept of the Internet of Things. It uses a “Smart” Gateway that converts existing NMEA data into Signal K, and its installation consists simply of wiring the gateway to NMEA0183 and/or NMEA 2000 networks and plugging the gateway into a wireless router [If properly designed, built, and maintained, a battery can provide many years of reliable service. A new battery might not initially provide 100% capacity. The capacity typically improves over the first few years of service, reaches a peak, and declines until the battery reaches its life limit. A reduction to 80% of the rated capacity is usually defined as the end of life for a lead-acid battery. Below 80%, the rate of battery deterioration accelerates, and it is more prone to sudden failure resulting from a mechanical shock or a high discharge rate. Note that even under ideal conditions, a battery is expected to eventually wear out . With the objective of maximum availability of the energy storage system, the storage capacity and the useful life of the batteries depends to a large extent on a suitable management of their use . OverloaThe dynamic behavior of battery operation has been studied by a large number of researchers . The modIn addition to these electrical models, other mathematical models have been developed to estimate the parametric variations depending on the values associated with the time of use/disuse and variations of the temperature of the battery . These mTo monitor the useful life of batteries, the aging phenomenon must be analyzed. Batteries age both when they are stationary  and when they are subjected to a cyclic operation (cycling ageing) . In the In practical terms, these effects translate into two repercussions: An increase in the internal resistance of the battery and a loss of capacity . Althougini) according to Equation (1).The most visible consequence of the aging of a battery is the gradual loss of its capacity, which is established through the parameter of the state of health of the battery (SOH). The SOH is calculated as the quotient between the current capacity (Cap) of the battery and the initial capacity Plate sulfationSoft and hard shortsPost leakageThermal runawayPositive-grid corrosionIt is known that batteries have different failure modes depending on their technology . Nowadayin, which in turn depends under normal conditions largely, but not exclusively, on the external temperature Tex due to weather conditions. The outside temperature, Tex, is “filtered” by the battery installation chamber and becomes the ambient temperature Tamb.Some of the failure modes listed above, especially dry out, positive-grid corrosion and thermal runaway, are strongly dependent on the increase in the internal battery temperature Tin has a strong influence on aging, grid-corrosion rates, and rates of water loss (dry out) due to evaporation or hydrogen evolution at the negative plates (self-discharge), which all increase with increasing temperature. On the other hand, a (moderate) temperature increase may improve service life in applications involving severe cycling [T cycling . Dry out is a phenomenon that occurs and is accelerated due to excessive heat  and over-charging, which can cause elevated internal temperatures, high ambient  temperatures, and contributes decisively to grid corrosion. Up to 82–85% of the failures exhibit signs of dry out . It is oamb, the Vfloat current, and the ripple current effect. In turn, the causes of the increase in the internal temperature of the battery are the ambient temperature TThermal runaway is a condition in which the battery temperature increases rapidly resulting in extreme overheating of the battery, therefore the battery can melt, catch on fire, or even explode. Thermal runaway can only occur if the battery is at a high-ambient temperature and/or the charging voltage is set too high ,60. Althamb in different ways. Among diverse batteries of different make and model, there are significant differences in the float voltages that can cause different aging periods. The following factors contribute to the response variation in each battery behavior: Chemistry of the battery and construction, age of the battery, and especially chamber conditions where batteries are installed [Different batteries can be affected by the float voltage and the ambient temperature Tnstalled .If the process of heat generation produced internally in the battery reaches an advanced uncontrolled phase, violent boiling will occur together with a rapid generation of gas causing in turn over-pressurization, which if not detected in time can cause catastrophic damage due to emissions of hydrogen, oxygen, hydrogen sulfide gas (an irritant), and atomized electrolyte. This process can cause a fire or explosion in the installation chamber . In thisRipple current is another contributor for battery inner temperature increase. Battery manufacturers recommend that under normal float-charge conditions, battery ripple RMS (root mean square) voltage must be limited to <0.5% of the Direct Current (DC) voltage applied to the battery. This ensures that the instantaneous cell voltage will not fall below the open cell voltage or rise above the maximum float-charge voltage. It also eliminates the consequential battery heating that would occur from constantly cycling the battery through discharging and recharging states. Many early laboratory and real-world studies of lead acid (Pb) have shown that Alternating Current (AC) ripple may cause the cell to experience shallow discharge cycles, that in turn may lead to gassing, grid corrosion and internal heat generation ,64.It is well known that high temperature is the “killer” of all batteries, and that its effect varies depending on the manufacturer and model or the type of technology used in its manufacture. Lead acid at 95 °F (35 °C) will experience a 50% shortened life, while Ni-Cd will have a 16%–18% shortening of life ,62. For At the other end, the low-temperature range slows down the internal chemical reactions in any battery. The degree of reduced performances vary according to the technology also. For example, at temperatures around freezing, a VRLA may need capacity compensation of 20%. The lead-calcium cell using 1.215 specific gravity acid will require a doubling of capacity, while the Ni-Cd will need about an 18% increased capacity. Under ideal conditions, the trend analysis of certain battery parameters, especially temperature, impedance, capacity, and SOH, would be an excellent tool to observe how the batteries degrade over time, and when a decision can be made to replace them. However, as mentioned above, easy access to all of these parameters is not always available ,66,67.In the data connection layer of the NMEA 2000 network, the main functions of the CAN interface are: Generating the linear stream of bits, controlling the access to the network, as well as the controlling of errors and automatic transmission of error messages.The CAN network is a linear, bit-orientated main. To avoid collisions and errors of transmission, the CSMA/CA method of the access to the main is applied, which determines that the reaction time of all drivers must be less than the transfer time of a bit .NMEA 2000 consequently inherited these limitations that initially conditioned its use in large vessels. Some of these limitations were the length of the main speed versus transmission speed; since all the CSMA/CA devices have to work with the same speed and in long lines, differences of a signal due to delays, may appear. Currently, this restriction can be solved by using NMEA 2000 network segments with devices such as network bus extenders (NBE) .NBE100 devices were developed to be able to extend the maximum node count up to 250, as well as the limit network trunk length and the cumulative drop length of a NMEA 2000 network. Without extenders a single network has a maximum of only nodes allowed, a network trunk length of 200 m, and a maximum cumulative drop length of 78 m.In 2001, NMEA published its NMEA 2000 standard and was also adopted as an IEC 61162-3 standard. More than 140 renowned companies belong to the manufacturer’s list of NMEA 2000 developer members. NMEA 2000 has had a great penetration de facto in medium and small length vessels. Later in 2012, NMEA announced the project OneNet  to develThe structure of NMEA 2000 is shown in In the NMEA 2000 network, devices are available for the measurement, monitoring, supervision and data processing of parameters from the navigation system, as well as graphical displays for the analysis of short-term trends. There are also devices for the registration of such data, such as the Voyage Data Recorder (VDR). These devices are reviewed in the following sections. Maretron’s DCM100  is a senBattery VoltageBattery CurrentRipple VoltageBattery Case TemperatureState of ChargeTime RemainingBattery CapacityBattery TypesCharging InefficienciesCharge Efficiency Factor (CEF)Discharging InefficienciesPeukert ExponentCharge Efficiency Factor CalculationIf “Battery” type is selected, many battery-related options become available , where dThe PB200 Weather-Station instrument is designed to output time series of data of weather parameters instantaneous values. Additionally, it is equipped with temperature and barometric pressure sensors that help to make trend analyses and forecasts on changing weather patterns . This wein [Maretron’s TMP100 module measures the temperature for up to 6 temperature probes and reports the information over an NMEA 2000 network . The TMPin .The Vessel Data Recorder VDR100  is a kinAdditionally, this activity can be shown in the form of the time series of data of all the parameters that the DCM100 is able to monitor. In turn, the NMEA 2000 network has the ability to display on-line mobile windows of up to 1-week amplitude, and integrates the ability to perform treatment of alarms and emergencies. The N2KView software  or a MarMaretron’s SMS100 short message service (Text) module is a mobile or cellular modem dedicated to sending alerts or alarm messages to selected phone numbers . This deTrend analysis is a quantitative technique that can be used to identify potentially hazardous conditions based on past empirical data obtained from time series of data. A trend analysis can reveal a movement toward unacceptable, undesirable, or dangerous reliability, safety, or levels of assurance. Its application dramatically decreases uncertainty and emergency replacements allowing to make adequate decisions for maintenance planning . In addiIn this work, a scenario under real weather conditions of navigation has been considered. This scenario is one in which a set of batteries are affected in terms of the dynamics of the temperature variable, by a set of numerical values associated with a time series of a stochastic process of seasonal character. t is the trend, St is the seasonal component, and It is the noise or random part.One way of describing time series is based on the idea of decomposing the variation of a series into several basic components, which becomes especially interesting when in the series a certain tendency or certain periodicity is observed . This deIn these conditions, a battery will experience changes associated with a dynamic in the form of random seasonal time series, that is, the temperature of the banks batteries will oscillate in successive diurnal warm-ups and nocturnal cooling, with higher average values in summer and lower in winter.The damage caused to a battery due to exposure to high temperatures is not reversible, even if subjected to a subsequent cooling. For example, the corrosion effect of the positive grid cannot be eliminated and occurs at all temperatures, which is simply a matter of the speed at which the corrosion process occurs. The only solution is to try to control and avoid as far as possible the causes involved in the increase of the temperature in batteries, with the criteria of economic profitability against the risks of failure .amb is made. From a general point of view, the method developed in this work is based on the performance of a comparative analysis of the behavior of the parameters presented in A difference introduced in the currently presented work is that the operation and tests of the batteries have been made in real navigation conditions, where the possibility of using refrigerated chambers for batteries installation is not feasible in most cases, from an economic point of view or from an energy consumption point of view.ex has on the dynamics of the internal temperature Tin of the batteries, and also on the tendency of the battery installation chamber’s ambient temperature Tamb. Both show a behavior whose dynamics are associated with random seasonal time series, influenced by the external temperature Tex, as can be verified by a cross-correlation analysis between the lagged series.One of the first characteristics observed in the analysis of the series recorded in the tests is the important influence that the external temperature Tin increase is due to the superposition of a set of parameters that overlap their effects as heat sources that increase the internal temperature of the battery. Note that Tex whose source lies in the existing weather conditions. The second parameter is the ambient temperature Tamb, which results from the “filtering” effect caused by the installation chamber on Tex. The third parameter is due to the contribution to the temperature increase produced by the ripple current (AC), whose source comes from the power electronics of the battery charger specifically associated to the aging of its components (diodes). The first parameter to consider is the external temperature Tfloat) contributes to the increase in temperature and the reduction of the internal impedance of the battery, which in turn causes the battery to accept even more current, that is able to trigger a process of thermal runaway instability after a previous and accelerated dry out process [The float input current  and battery installation chamber temperature (Tamb). The objective is to apply predictive fault diagnosis using early detection of the failure or loss of capacity of the battery heat dissipation process to prevent any undesirable increase in temperature, in order to avoid the development of failure modes such as dry out and thermal runaway. Taking into account that dry out is often a prior effect to other failure modes such as grid corrosion [This work focuses on proposing a specific method for the comparative trend analysis of the internal temperature of the batteries , the materials used, the size of the heat dissipation areas, and the ventilation conditions, etc., and that affects the batteries in a variable way depending on the type of batteries and their intrinsic aging state condition.in and Tamb is observed, it becomes a prior indicator that the battery is not being able to dissipate normally the heat generated inside. In other words, this first failure diagnosis of the heat dissipation process becomes a predictive indicator to avoid possible dry out and eventual thermal runaway failures.If an increasing divergence through an additional comparative trend analysis of both Tex are shown. In the dynamic behavior of the time series, it is easy to recognize their seasonal and random nature, especially in the case of the external temperature series. In both Figures, the temperatures of the battery banks are compared to the outside temperature, observing a follow-up with a certain time delay.The results of the parametric measurements made and their interpretation are shown in this section. in and external temperature Tex have certain differences in their appearance, mainly due to the higher frequency components of Text, while, Tin shape is smoothed. This is due to the large thermal inertia of the battery. It takes time for the battery to absorb temperature and it takes time for the battery to relinquish temperature. In addition, the daily cyclic behavior of all data series is completely evident.It should be realized that battery temperature Tex and the internal temperature Tin of the battery are correlated with a certain delay value, as affirmation of the working hypothesis.in temperature series of the three battery banks are shown, where maximum coincidence in the dynamic evolution of the house and engine banks exist, because in this case, they share the same installation chamber, while the bow thruster battery bank are installed in a different chamber in the bow vessel area. In all cases, it is evident that the thermal inertia and the enclosures of the installations act as smoothing filters of the battery inner temperature series, notably suppressing the high-frequency random component observed in the outdoor temperature series (Tex) in In The house battery bank presents in general an increase of temperature with respect to the engine battery bank and the bow thruster battery bank, which is justified by the fact that it normally supplies a permanent power consumption to electrical and electronic equipment. Requests for energy supply from the other two banks can be described as sporadic, compared to those made to the house battery bank.In In The peaks of energy demand (in green, in and Tamb temperatures of the house battery. In conditions of normal operation it is observed that the curve of the installation chamber temperature Tamb is lifted upwards from the curve of the internal temperature Tin of the house battery. This is explained by the activation of the heat dissipation process from the inside of the battery to the outside in the installation chamber, which increases Tamb. Later when the current demand stops, Tamb tends to overlap progressively with the internal temperature Tin.in and Tamb, with a high correlation index and with a barely significant delay between both curves.In in, the temperature of the battery installation chamber Tamb, the floating charge voltage of the battery Vfloat and the float current Ifloat. Under normal operating conditions, as shown in the laboratory test of in and Tamb temperature curves is significant, whereby the heat dissipation process of the battery bank is working adequately, taking into account the specific characteristics and conditions of the type of battery and the installation chamber.In in is lower than Tamb, and in curve. This results in the crossing of the curves and an increasing divergence between Tin and Tamb.Initially In this case, the progressive divergence of the two curves shows that the heat dissipation process capacity is being exceeded and, if this condition persists, the development of the dry out phenomenon and eventually the thermal runaway occurrence is predictable.For safe maritime navigation, having a monitoring system based on observing the actual condition of use and specific technical characteristics of the ship’s energy storage system is especially important and critical. The NMEA 2000 network—together with intelligent sensor devices for battery monitoring, temperature measurements, ship data logging, and associated software developed for data processing—allows for obtaining time series of data and for the application of trend analysis for better management of alarms, in order to face the appearance of incipient and/or sudden failures.The predictive diagnostic method proposed in this study does not depend on a temperature threshold value, but on the previous failure of the heat dissipation process. This behavior demonstrates that the working hypothesis proposed in the comparative trend analysis can be used as a recognition pattern for the predictive diagnosis of eventual dry out and thermal runaway failures.In the opinion of the authors, the equipment that is usually installed for the supervision of battery banks in navigation applications offers sufficient information on the most important parameters of battery operation. However, from the point of view of the implementation of predictive diagnostic techniques, the information they offer is not always easy to analyze by potential users who are largely not experts in the field, and much of its information potential is lost.For truly useful decision making in critical situations, it would be desirable that the information is oriented toward the mode of failure and displayed with multi-parametric high level integration. In that way, comparative trend analyses and correlation of the involved parameters could be applied. Even more, this information should be given in the form of patterns of immediate or easy recognition, that reliably allow the detection of an assignable cause such as a failure of the heat dissipation process of the battery, useful for predictive dry out and thermal runaway fault diagnosis. It is concluded that these failure modes can be conveniently diagnosed by easy recognized patterns obtained performing comparative trend analysis to the variables measured onboard.


Vs) has advanced our understanding of their pharmacology. Herein, we report the purification and characterization of a novel non-selective mammalian and bacterial NaVs toxin, JZTx-14, from the venom of the spider Chilobrachys jingzhao. This toxin potently inhibited the peak currents of mammalian NaV1.2–1.8 channels and the bacterial NaChBac channel with low IC50 values (<1 µM), and it mainly inhibited the fast inactivation of the NaV1.9 channel. Analysis of NaV1.5/NaV1.9 chimeric channel showed that the NaV1.5 domain II S3–4 loop is involved in toxin association. Kinetics data obtained from studying toxin–NaV1.2 channel interaction showed that JZTx-14 was a gating modifier that possibly trapped the channel in resting state; however, it differed from site 4 toxin HNTx-III by irreversibly blocking NaV currents and showing state-independent binding with the channel. JZTx-14 might stably bind to a conserved toxin pocket deep within the NaV1.2–1.8 domain II voltage sensor regardless of channel conformation change, and its effect on NaVs requires the toxin to trap the S3–4 loop in its resting state. For the NaChBac channel, JZTx-14 positively shifted its conductance-voltage (G–V) and steady-state inactivation relationships. An alanine scan analysis of the NaChBac S3–4 loop revealed that the 108th phenylalanine (F108) was the key residue determining the JZTx-14–NaChBac interaction. In summary, this study provided JZTx-14 with potent but promiscuous inhibitory activity on both the ancestor bacterial NaVs and the highly evolved descendant mammalian NaVs, and it is a useful probe to understand the pharmacology of NaVs.Exploring the interaction of ligands with voltage-gated sodium channels (Na V) is composed of a pore-forming alpha subunit and one or two covalently or non-covalently associated beta subunits. The alpha subunit has a topological structure of 24 transmembrane segments (TMs), which could be further divided into four homologous domains, with each domain containing six TMs. A total of nine NaV subtypes have been identified in mammals with different tissue distributions and accordingly diverse functions: the NaV1.1–1.3 subtypes that are mainly located in the central nervous system (CNS); the NaV1.4–1.5 subtypes that are expressed in the skeletal muscle cells and the cardiac myocyte cells, respectively; the NaV1.7–1.9 subtypes that are mostly restricted to the peripheral nervous system (PNS); and the NaV1.6 channel, which is expressed in both the CNS and the PNS [Vs were expressed in non-excitable cells such as astrocytes, microglia, macrophages, and cancer cells, and functioned in a non-canonical way [+ selectivity mechanism of mammalian NaVs are yet to be explored, and the milestone progresses regarding the cryo-EM structures of eukaryotic NaVs such as cockroach NaVPas [V1.4 [V1.4 [Vs, and are rich mines for developing drugs for treating NaV-related diseases and discovering probes for NaV researches [Vs; among these, site 1, site 3, site 4, and site 6 are binding sites for small disulfide-rich peptides from animal venoms [V1.2–1.8 channels, it is difficult to purify NaV subtype-specific modulators from animal venoms. This is because that these toxins might bind to NaVs by recognizing a common structure in them, which is similar to that of tetrodotoxin (TTX) occluding the pore of TTX-sensitive NaVs  by recognizing the conserved structures in them [V subtypes with comparable affinity are rare, as there are substantial differences between them. Furthermore, NaV voltage sensor binding peptide toxins (site 3 and site 4 toxins) have always shown reversible and state-dependent binding property. The identification of toxins deviating from these rules would advance our understanding of the pharmacology of NaV channels.The mammalian voltage-gated sodium channel  template for designing drugs targeting human NaVs [Vs, relatively few peptide toxins acting on bacterial NaVs have been reported. Mammalian Naell line . Since tian NaVs ,16. Thisreported ,21,22,23ic level . The cryevel [Vs . Further in them ,27. In toflurane ,28. Addiman NaVs . HoweverChilobrachys jingzhao, is purified and characterized. It showed promiscuous inhibitory activity on mammalian NaV1.2–1.8 channels and the bacterial NaChBac channel, and also inhibited the fast inactivation of the NaV1.9 channel. Kinetics data and mutation analysis proved that JZTx-14 was a gating modifier of both mammalian and bacterial NaVs. However, its irreversible blocking effect on mammalian NaV and its state-independent binding with the channel suggested that the action mode of JZTx-14 might be different from those classical site 4 toxins: that is, JZTx-14 might stably bind to a conserved toxin pocket deep within the NaV1.2–1.8 voltage sensor regardless of channel conformation change, however, its inhibitory effect on NaVs requires the toxin to trap the S3–4 loop in its resting state. This study provided a novel toxin to understand the pharmacology of both the mammalian and bacterial NaVs.In the present study, a novel peptide toxin, JZTx-14, from the venom of the spider C. jingzhao, JZTx-14, which non-selectively and potently inhibited NaV1.2–1.8 currents. This toxin had a retention time of 40.8 min in the RP-HPLC purification of the venom  (https://blast.ncbi.nlm.nih.gov/Blast.cgi) matched the toxins JZTx-14 and Jingzhaotoxin F4-32.60 (Chilobrachys jingzhao matched JZTx-14 with the highest score (UniProtKB/Swiss-Prot accession number: B1P1C0.1), with an identification coverage ratio of 100%. 1-C2-C3C4-C5-C6), and the mode of the disulfide bonds might be C1-C4, C2-C5, and C3-C6 (the number indicates the relative position of cysteine in the sequence). Sequence alignment showed that JZTx-14 had low identity with the peptide toxins in the database  C. AdditiF4-32.60 D. JingzhF4-32.60 D. We anadatabase F.Vs NaV1.2–1.9 heterologously expressed in HEK293T or ND7/23 cells. The NaV1.8 and NaV1.9 channels were chimeras, as described in our previous studies [V1.2–1.7 currents , 426.3 ± 48.8 nM for NaV1.3 (n = 5), 290.1 ± 23.2 nM for NaV1.4 (n = 6), 478.0 ± 32.0 nM for NaV1.5 (n = 5), 158.6 ± 29.4 nM for NaV1.6 (n = 4), 188.9 ± 46.3 nM for NaV1.7 (n = 6), and 824.0 ± 68.7 nM for NaV1.8 (n = 5)  S3–4 loop with that of NaV1.9, and tested its response to JZTx-14. The data showed that 2 µM toxin only slightly inhibited its peak currents, but profoundly slowed its fast inactivation ; all of these mutants showed no significant change of their sensitivity to JZTx-14, and an alanine scan mutation analysis of the DII S3b–S4 paddle motif (from the 795th glutamine to the 808th arginine) did not identify any key site (data not shown). We propose that JZTx-14 associated with the DII S3–4 loops of NaV1.2–1.8 by binding multiple amino acids in this region. It well explained the targets proximity of JZTx-14 among NaV subtypes.We tested the activity of JZTx-14 on mammalian Na studies ,31. Thei current H. The do (n = 5) I,J. Thestivation K. This sV1.2 was used as a representative channel and the effect of JZTx-14 on its activation was tested. As shown by the traces in V1.2 currents at every depolarizing voltage, and the toxin did not change its peak current voltage or reversal voltage (n = 10). Additionally, we compared the I–V relationships by normalizing the currents in the toxin group to their maximum peak current . The data showed that the curves superimposed between the depolarizing voltages of −50 mV to +50 mV, while the proportion of opening channels at voltages of +60 mV to +100 mV in the toxin group was higher than that in the control group . We propose that JZTx-14 trapped the NaV1.2 DII voltage sensor in one of the deactivated states. We further conducted the toxin dissociation assay to explore the state-dependent binding of JZTx-14 with NaV1.2. As shown by the voltage protocol in V1.2 currents by binding to the DII voltage sensor. Saturating doses of JZTx-14, HNTx-III, and TTX fully blocked the NaV1.2 currents elicited by t1. However, in contrast to a large recovery of the NaV1.2 current in t2 in the HNTx-III group, no current recovery was observed in the JZTx-14 and TTX groups (n = 3–5). It is reasonable that strong depolarization could not cause TTX dissociation from NaV1.2 as its action mechanism. The action mode of JZTx-14, which is characterized by its state-independent binding with NaV1.2 and the irreversible blocking of NaV1.2 currents, is distinct from other toxins acting on the NaV DII voltage sensor, such as HNTx-III and HWTx-IV [Nacurrents D. These  HWTx-IV ,33.V, NsvBa, was relatively weaker, with 1 µM toxin inhibiting its currents by 46.8 ± 3.3% (n = 6). The dose–response curves showed that the IC50 values of JZTx-14 for the NaChBac and NsvBa channels were 320 ± 38 nM and 1400 ± 200 nM, respectively (n = 5–7). For the bacterial NaVs NaVPz and NaVSP, 1 µM JZTx-14 only caused a 34.5 ± 6.5% and 25.7 ± 5.7% inhibition of their currents, respectively (n = 3). JZTx-27 is another NaChBac and NsvBa channel antagonist characterized in the same venom [Vs. However, compared with JZTx-27, JZTx-14 showed more potent activity on mammalian NaVs. As for mammalian NaVs, we tested the effect of JZTx-14 on the steady-state activation and inactivation of NaChBac. As shown in a = −33.8 ± 0.41 mV and –23.7 ± 0.98 mV, Ka = 6.2 ± 0.35 mV and 9.9 ± 0.87 mV, for control and toxin-treated channels, respectively; n = 5). Furthermore, the toxin positively shifted the steady-state inactivation relationship of NaChBac . These data suggest that JZTx-14 acted on NaChBac as a gating modifier.As shown in 8 ± 3.3% B, n = 6.me venom . In the Vs determined this loop in NaChBac to be 103–FAGAQFV–109 , 832.6 ± 42.1 nM for F103A (n = 7), 653.3 ± 92.1 nM for G105A (n = 5), 809.8 ± 87.5 nM for Q107A (n = 6), 3472.3 ± 195.7 nM for F108A (n = 6), and 1367.3 ± 129.3 nM for V109A (n = 5) mutant channel, respectively. We further tested the effect of JZTx-27 on the F108A mutant channel, and the IC50 was determined as 116.8 ± 11.0 nM, with only a ~2.5-fold change being observed when compared with wild NaChBac (IC50 = 46.7 ± 1.9 nM) (n = 6–7). These data suggest that JZTx-14 and JZTx-27 bind to different key residues in NaChBac, although their interacting surfaces in the channel might partially overlap.Our previous study showed that JZTx-27 bound with the S3–4 extracellular loop of NaChBac, with the 103th phenylalanine (F103) being the key residue . We analAQFV–109 H,I. The C. jinzhao as a broad spectrum NaVs toxin that acted on mammalian NaV1.2–1.9 subtypes and the bacterial NaChBac channel. It showed multiple phenotypes in tested NaVs, with the toxin inhibiting the fast inactivation of the NaV1.9 channel and mainly inhibiting the peak currents of NaV1.2–1.8 and NaChBac. For NaV1.2–1.7 subtypes, the toxin also slightly inhibited their fast inactivation. The promiscuous mammalian NaVs inhibitory activity of JZTx-14 resembled that of GrTx1 and GsAF1 [V1.5 channel. JZTx-14 acted on mammalian NaVs and the NaChBac channel as a gating modifier and the DII S3–4 loops of mammalian NaVs and the NaChBac S3–4 loop were involved in toxin association; however, the action mode of JZTx-14 on mammalian NaVs was a departure from classical site 4 toxins. We suggest that JZTx-14 might be useful in exploring the pharmacology of NaVs as follows.This study purified and characterized JZTx-14 from the venom of nd GsAF1 . HoweverV channel to inhibit both the activation and fast inactivation of the NaV1.2–1.7, especially for the NaV1.2 and NaV1.3 channels. This is not surprising, as several toxins such as ProTx-I, ProTx-II, and TsVII were shown to interact with multiple regions in NaVs [V1.2–1.8 channels, its primary and high-affinity binding site in them might be site 1 or site 4 [V pore [V1.2 channel in V, and JZTx-14 resembled KIIIA in irreversible binding with NaV [V DII voltage sensor and trapping it in the deactivated state. Additionally, the NaV1.5/1.9DIIS3–4 chimeric channel clearly showed that the DII S3–4 extracellular loop is involved in JZTx-14 association. However, the observations that JZTx-14 irreversibly and state-independently blocked NaVs were departures from the action mode of classical NaV site 4 toxins, such as HNTx-III and HWTx-IV [V DII voltage sensor could be easily washed off by bath solution and easily dissociated by voltage sensor conformation change, both of these were not observed in our experiments. Moreover, the irreversible binding of JZTx-14 to NaV1.2–1.8 could not be simply explained by its potential membrane-binding capability, as the action of JZTx-14 on the NaChBac channel was reversible. It is possible that JZTx-14 embedded into the membrane and bound to a deep region in the voltage sensor of NaV1.2–1.8 by recognizing a common structure in these channels; however, this binding might be silent, and the toxin inhibitory effect on NaVs requires the participation of the S3–4 extracellular loop. We raised a model that JZTx-14 seated in a toxin pocket that was deep within the voltage sensor, and used another surface to interact with and trap the S3–4 loop in its deactivated state from the inside and accordingly impeded the channel activation. This model needs to be validated by more solid data in future studies, and differs from that of the proposed action mode of classical site 4 toxins such as β-scorpion toxin CssIV, in which the toxin binds to the S1–2 and S3–4 loops cleft from the extracellular side [In this study, JZTx-14 seemed to bind to two sites in mammalian Na in NaVs ,36. As Jr site 4 . Guanidi [V pore ,38. We cwith NaV , the fac HWTx-IV ,33. As ilar side .Vs were inactivated via a C-type inactivation mechanism, in which the collapse of the selectivity filter served as an inactivation gate [The mutation study showed that JZTx-14 acted on NaChBac by binding to the S3–4 extracellular loop, with the 108th phenylalanine (F108) being the key residue. This action mode resembled the previously reported NaChBac channel toxin JZTx-27, although their key residues in the channel were different . We specion gate . The staion gate . The datVs were deemed to be ancestors of mammalian 24TMs voltage-gated calcium and sodium channels [Vs and NaChBac [Vs, and it was assumed that the isoflurane binding sites in NaChBac and mammalian NaVs are conserved [Vs with similar affinity, but with a distinct mechanism of isoflurane. Whether or not the toxin used the same toxin surface for interacting with these two disparate types of channels remains to be explored.The bacterial Nachannels ,45. Sevechannels . Additio NaChBac . Furtheronserved . In thisC. jingzhao was collected by an electric stimulation method as previously described [2O to a final concentration of 5 mg/mL immediately before being subjected to RP-HPLC purification. The first round of semi-preparative RP-HPLC purification was performed in a Hanbon HPLC system  by using a C18 column  and a 45-min linear acetonitrile gradient from 10% to 55% at a flow rate of 3 mL/min. The fraction containing JZTx-14 was collected, lyophilized, and subjected to the second round of RP-HPLC purification in Waters 2795 HPLC system  by using an analytic C18 column  and a 24-min linear acetonitrile gradient from 20% to 44% at a flow rate of 1 mL/min. The JZTx-14 store solution was made by dissolving lyophilized toxin in sterile ddH2O, and the toxin concentration was determined using the Enhanced BCA Protein Assay Kit following the manufacturer’s instruction . The standard curve of the assay was created using synthetic HWTx-I toxin of known concentration. The N-terminal 11 amino acid sequence of JZTx-14 was determined by Edman degradation in an automatic protein sequencer .The venom of the spider escribed . The colTM 5800 system . Briefly, 1 µL JZTx-14 sample solution was mixed with 1 µL saturated CCA (α-cyano-4-hydroxycinnamic acid) solution, pointed onto a sample plate, and then subjected to mass spectrometric analysis in positive reflectron mode. The initial laser intensity was set to 3800, and was finely adjusted to obtain a good resolution and signal-to-noise ratio. Mass calibration was achieved using an external standard. We determined the C-terminal sequence of JZTx-14 by combining chymotrypsin digestion and LC-MS analysis in a Q ExactiveTM mass spectrometer . Briefly, 10 µg JZTx-14 was reduced, alkylated, and digested with chymotrypsin and subjected to LC-MS analysis. The data was searched against the venom gland cDNA library database of Chilobrachys jingzhao by using MaxQuant.The purity and molecular weight of the purified JZTx-14 was analyzed in an AB SCIEX TOF/TOFVs NaChBac, NsvBa, NaVPz and NaVSp were from professor David E Clapham lab , and were cloned in a pTracer-CMV2 vector. The mammalian NaV cDNA clones (NaV1.2–NaV1.8) were from professor Theodore Cummins lab , and were cloned in the pCDNA3.1 or pCMV-blank vectors. The NaV1.8/1.7L5 channel and the NaV1.9-EGFP channel were as described in our previous studies [E. coli DH5α chemical competent cells. All of the mutants were sequenced to ensure that the correct mutations were made. The CHO-K1 cells and HEK293T cells were used for bacterial NaVs and mammalian NaVs NaV1.2–NaV1.8 heterologous expression, respectively. Cells were cultured under standard conditions  in a humidified incubator and transfection was performed when cells reached 80–90% confluence. All of the transfections were performed using Lipofectamine 2000 following the manufacturer’s instructions . Four to six hours after transfection, cells were seeded onto PLL-coated coverslips. Twenty-four hours after transfection, cells were ready for patch-clamp analysis. ND7/23 cells were used for NaV1.9-EGFP chimeric channel expression and the conditions were as previously described [The cDNA clones of bacterial Na studies ,31. MutaV currents, the bath solution contained (in mM): 140 NaCl, 2 CaCl2, 1 MgCl2, 5 KCl, 10 glucose and 20 HEPES (pH = 7.3); the pipette solution contained (in mM): 140 CsF, 1 EGTA, 10 NaCl and 10 HEPES (pH = 7.3). TTX was added to bath solution to a final concentration of 1 µM when recording NaV1.9 currents. Unless otherwise indicated, all chemicals were products of Sigma-Aldrich . After breaking in, the serial resistance was controlled to be less than 10 MΩ, the voltage error was minimized by using 80% serial resistance compensation, and the speed value for compensation was 10 µs. To minimize the fast capacitance, only the tip of the pipet was filled with pipet solution, and the artificial capacitance effect was canceled by using the computer-controlled circuit of the amplifier. Data were acquired by the PatchMaster software  and analyzed by Sigmaplot 10.0 , Igor Pro 6.10A  and Graphpad Prism 5.01 .Whole-cell current recordings were performed in an EPC10 USB patch-clamp platform . The recording pipets were prepared from glass capillaries (thickness = 0.225 mm) with a PC-10 puller , and the pipet resistance was controlled to be 1.5–3 MΩ. For recording Na1/2, V and K represent the midpoint voltage of kinetics, the test voltage and the slope factor, respectively. The dose–response curves were fitted by a Hill logistic equation to estimate the potency (IC50) of the toxin.Data were presented as mean ± SEM. N was presented as the number of separate experimental cells. The G–V and SSI curves were fitted by a Boltzmann equation:


The optimal technique for airway management in patients with cervical pathology remains unclear. Intubating laryngeal mask airway devices such as LMA CTrach and LMA Fastrach have not been compared for cervical spine (C-spine) movements in the context of cervical pathology. The present study aimed to determine upper C-spine movements by radiography during intubation with different devices as well as comparing the duration and success of intubation in cervical surgery.2 were excluded. Participants were randomized to one of the 3 groups: LMA CTrach, LMA Fastrach, or the Macintosh laryngoscope. C-spine motion was evaluated by measuring angles created by bordering vertebrae at cervical 1/2 and 2/3  segments on 2 lateral cervical radiographs for each patient. Intubation time, ease of intubation, number of attempts, and success rate were also documented. Sixty patients scheduled for elective cervical surgery were registered in this prospective, randomized study. Patients with cervical trauma/injury, previous neck surgery, and body mass index (BMI) of >35 kg/mDemographic data were similar in all the groups. The cervical movement with LMA CTrach and LMA Fastrach compared to the Macintosh laryngoscope were similar at C1/2. However, LMA CTrach significantly reduced extension compared to LMA Fastrach and Macintosh laryngoscopes at C2/3. Duration of intubation was significantly shorter with the Macintosh laryngoscope. The rate of successful intubation was 80% with LMA Fastrach and 100% with both LMA CTrach and the Macintosh laryngoscopes.The LMA CTrach laryngoscopy involves less upper C-spine movement than the LMA Fastrach and does not increase the duration of the intubation period.  Intubation with various airway devices causes cervical spine (C-spine) extension to some degree. The process gains importance especially in emergency situations with cervical injury and in C-spine surgeries . The main concerns of anesthesiologists for airway management both in cervical injury and C-spine surgeries include avoiding prolonged intubation time and preventing neurologic damage due to excess cervical movements .Conventional laryngoscopy with a Macintosh blade remains the most familiar way to enable tracheal intubation. However, maneuvering for intubation and adjustment of the oropharyngeal and laryngeal axes produces C-spine movement [4]. In a recent systemic review, alternative intubation techniques performed in patients with cervical immobilization were compared with the Macintosh laryngoscopy [5]. The authors concluded that evidence of the efficacy of alternative devices was missing. Fiberoptic intubation is still the most ideal technique to secure an airway in patients with predicted difficult intubation [2]. Fiberoptic laryngoscopy is considered to facilitate the least cervical movement during laryngoscopy but has several limitations like requiring a cooperative patient and lasting a long time, making it unsuitable for emergencies . However, as intubating laryngeal mask airways (ILMAs), LMA CTrach and LMA Fastrach are alternative techniques that may be useful when fiberoptic bronchoscope is not available. ILMAs have been validated for ventilation and as a conduit to tracheal intubation in patients with difficult airways. LMA CTrach was developed from LMA Fastrach with additional advantageous features like visualizing the glottis and intubation process . However, both devices require experience to operate and the administration may prolong intubation time. Previous radiographic and fluoroscopic studies, purporting to evaluate C-spine movement during intubation with various intubating techniques, were carried out. . Intubating laryngeal mask airways have been compared for the success of tracheal intubation . However, there have been no studies investigating C-spine movements using both ILMAs. Moreover, the effects of different intubation techniques on cervical movements were followed among healthy subjects in nonemergency situations not including neck, throat, or cervical surgeries in patients with cervical immobility due to manual in-line stabilization or cervical collars and in cadaveric models in most studies . Comparative studies performed on patients with cervical pathologies who undergo cervical surgery are lacking in the literature. Therefore, this prospective, randomized radiographic study was conducted to compare the movements of the upper C-spine  during laryngoscopy via LMA Fastrach, LMA CTrach, and Macintosh laryngoscopes in patients with lower cervical pathology undergoing C-spine surgery. The secondary outcomes were the comparison of the intubation success and the duration. The ethics committee of the School of Medicine of Erciyes University approved this prospective, randomized, and controlled study (reference number: 2011/177). All the patients were informed about the study and written consent was obtained. Inclusion criteria were American Society of Anesthesiologists (ASA) physical status of I–III, ages between 18 and 70 years old, and patients undergoing elective C-spine surgery. Patients with documented cervical trauma or injury, previous neck surgery, body mass index (BMI) of >35 kg/m2, the possibility of pregnancy, or failed tracheal intubation (more than 2 intubation attempts with a device) were excluded. All patients’ preoperative height, weight, ASA physical status, BMI, and Mallampati scores were documented. Electrocardiography, noninvasive blood pressure measurement, and pulse-oximeter were monitored in all patients in a standard fashion. Before anesthesia induction, the patients were positioned in a neutral position. Following preoxygenation, anesthesia was induced with intravenous (IV) 2 mg/kg propofol, 1 mg/kg lidocaine, 0.5 µg/kg remifentanil, and 0.6 mg/kg rocuronium, and maintenance was achieved with sevoflurane in an air-oxygen mixture and IV infusion of remifentanil. In the operating room, 63 patients were randomly assigned by a computer random number generator to one of the 3 groups, corresponding to the 3 airway devices: Macintosh laryngoscope (Group M), LMA Fastrach (Group F), or LMA CTrach (Group C). We did not stabilize the head and neck or apply cricoid pressure. In Group M patients, conventional direct laryngoscopy was performed. The diameter of the endotracheal tube (ETT) was 7 or 7.5 mm in females and 8 mm in males. In Group F, size 3, 4, or 5 LMA Fastrach  depending on the weight of the patient was applied in accordance with the manufacturer’s instructions. A silicone reinforced ETT was lubricated and inserted. If the right position of the tube was achieved without resistance, the ETT was advanced into the trachea and the cuff was inflated. The position of the ETT was determined by auscultation and capnography. Subsequently, a special stabilizing rod was used to remove the LMA Fastrach. In Group C, size 3, 4, or 5 LMA CTrach  was preferred depending on the weight of the patient. Initially, LMA CTrach was inserted without the viewer on, just as LMA Fastrach was applied. The cuff was inflated and the patient was ventilated. The viewer was then attached to the connector while holding the handle. When a clear image of the glottis and vocal cords was achieved, an ETT was inserted and intubation was visualized, and the ETT cuff was inflated. The viewer was then detached, and LMA CTrach was removed following the same procedure as for LMA Fastrach. The same experienced anesthesiologist, who had previously performed at least 100 intubations with each of LMA CTrach and LMA Fastrach and 500 intubations with the Macintosh laryngoscope, performed all laryngoscopies in order to minimize interoperator variability. The intubation techniques were recorded with a portable X-ray machine. Two recordings were performed a steady distance from the patient and the tube in the lateral position. The first was taken in a neutral position before the intubation process and the second was taken when the best view of the glottis was achieved with the LMA CTrach and Macintosh laryngoscopes. For the LMA Fastrach, if no resistance was felt as the ETT was advanced through the mask aperture into the trachea, it was thought to be at correct tube positioning and indicated time for the fluoroscopy. The movements of the cervical 1-2 and 2-3 segments were evaluated in radiographs. First we drew a reference line, which follows the cervical 2 dorsal alignment. Then we drew two more lines, which transected the driven reference line, the first one between the anterior and posterior arches of the cervical 1 and the other one through the cervical 3 basal plate of C3. Thus, there were two angles observable: one between the reference line and the arches of cervical 1, named alpha (α), while the second angle, named beta (β), was located between the reference line and the line passing through the cervical 3 basal plate. The lines were drawn and an investigator, who was unaware of the study group assignments, randomly measured angles using a goniometer in degree (°) unit. The duration of intubation, the number of attempts, and the intubation success rate were recorded. The duration of the intubation was recorded between the passage of the intubation device through the lips and inflation of the tracheal cuff. Intubation was considered successful if the patient was intubated in less than two attempts and failed in case of more than two. Estimated from the data of Watts et al. [15] (12.9 ± 2.1° extension) and based on the assumption of α = 0.05, β = 0.8 to detect 15% reduction in the movement of the upper C-spine, each group would need to include at least 18 patients. Therefore 20 patients per group were planned to be enrolled due to the probability of a 10% drop-out rate. SPSS 15.0 for Windows  was used for the statistical analysis. Data are expressed as mean ± standard deviation (mean ± SD), median (min–max), or numbers (n) and percentages (%). Discrete variables  were compared using the chi-square test. For numeric parameters of between-group comparisons, one-way analysis of variance (in the case of parametric test conditions) or the Kruskal–Wallis tests (if parametric test conditions could not be obtained) were used. Multiple comparisons of the Kruskal–Wallis test were done by Mann–Whitney U test with Bonferroni correction. Analysis of variance was used for repeated measures of arterial pressure, heart rate, and oxygen saturation and the angle measurements for both between-group and intergroup comparisons. P < 0.05 was considered statistically significant.Sixty-three patients were allocated to the intervention groups. For one patient, intubation with LMA Fastrach was not possible despite adequate manipulation, even after the third attempt. Following 3 esophageal intubations, correct intubation was achieved by direct laryngoscopy and this patient was excluded from the study. In Group C, the vocal cords of 1 patient could not be visualized despite all maneuvers and the patient was also excluded. One of the patients’ radiographic images could not be printed out so the angle measurement could not be analyzed in Group M. Subsequently, 20 patients per group were analyzed.2, remained stable in all groups and anesthesia was uneventful in all patients. None of the patients had hypertension or tachycardia as a response to laryngoscopy. The demographic data, Mallampati scores, and ASA physical status classification (P > 0.05) were similar in all groups (Table 1). The hemodynamic and ventilation parameters, such as arterial blood pressure, heart rate, oxygen saturation, and end-tidal COCervical alpha (α) angles in degrees at the C1/2 segment of the study patients are shown in Table 2. Baseline measurements were significantly different in Group M compared with the other groups (P = 0.004). According to the neutral baseline position, angulation of the C1/2 segment (α angle) decreased during intubation in Group F (P = 0.042) and in Group M (P = 0.001), whereas there was no significant difference in Group C (P = 0.159). The mean degree of the change in angulation compared with the preinduction baseline values at C1/2 1.2°, 1.1°, and 2.9° for Groups F, C, and M was not statistically significant.Cervical beta (β) angles of the studied patients are displayed in Table 3 in degrees. At the C2/3 segment (β angle) Groups F and M similarly showed a significant increase  in cervical motion, while there was no difference in Group C during intubation compared with the neutral position. The mean change in β angle during intubation was prominent in Groups F and M, but in Group C the extension was statistically less .The mean intubation time was 32 s in Group M, ranging from 10 to 120 s. It took the longest time to intubate patients in Group F and the shortest in Group M (P < 0.001) (Table 4). The mean duration of intubation was 98.8 and 61.0 s for the LMA Fastrach and LMA CTrach, respectively (P < 0.001). The numbers of patients intubated in the first attempt were 18 with the Macintosh laryngoscope, 15 with LMA CTrach, and 11 with LMA Fastrach while two attempts were required for 2, 5, and 5 patients, respectively (Table 4). The number of attempts significantly differed in Group F compared to Group M (P = 0.016). It required more than two attempts and was successful after repositioning the LMA Fastrach in 4 patients. The rate of successful tracheal intubation was 80% with LMA Fastrach and 100% with LMA CTrach and Macintosh laryngoscopes. Group F had the statistically lowest success rate (P = 0.009).The main result of this study is that the LMA CTrach significantly reduced extension compared to LMA Fastrach and Macintosh laryngoscopes at the C2/3 segment (β angle) without prolonging intubation time in patients undergoing elective C-spine surgery. According to our hypothesis, both LMA Fastrach and LMA CTrach would be associated with less cervical movement than Macintosh laryngoscopes, which was at least partially confirmed with LMA CTrach for the β angle. A study by Sawin et al. investigated the behavior of the intact C-spine during direct laryngoscopy with a Macintosh blade and proved the general assumption that a majority of cervical motions associated with laryngoscopy occur in the upper cervical region [4]. Subaxial segments (under C2) were displaced minimally. Thus, the present study was undertaken to quantify the motion of cervical segments 1/2 and 2/3. The times for radiography were chosen as once before induction and once during laryngoscopy immediately prior to insertion of the endotracheal tube or when the best view of the glottis was achieved as Hindman et al. reported maximal intubation biomechanics occurring at that stage [16]. A cadaver model of cervical instability found that supraglottic airways  caused less or equal C-spine movement compared to the conventional laryngoscopes  [9]. The authors therefore suggested that, due to the ease of training, supraglottic airways could be preferred in cervical trauma patients. Komatsu et al. tested ILMAs for controlling the airway in patients undergoing C-spine surgery who were wearing rigid cervical collars to simulate C-spine injury and found ILMAs a reasonable alternative for facilitating intubation [17]. Panjabi et al. defined the upper limits of the physiological motion as a rotation of over 20° in the sagittal plane [18]. The maximum cervical motion at C2/3 in our study was 15° for the Macintosh laryngoscope, 12.4° for LMA Fastrach, and 7.6° for LMA CTrach. In a video-fluoroscopic study, Sahin et al. observed a maximum movement of 18.5°, 16.7°, and 8.1° during direct laryngoscopy, intubation with ILMA, and fiberoptic laryngoscopy at C1/2 [6]. The maximum cervical motion produced with LMA Fastrach in both studies was close to the angle of the Macintosh laryngoscope. However, it was still lower than the instability limits argued by Panjabi et al. [18]. Even though the extension of the motion produced by LMA CTrach in the present study seems to be less than that produced by fiber optic laryngoscopy in Sahin’s study, it is actually difficult to compare the data of these similar studies. In the present study there was a significant difference in baseline angle measurements at C1/2. We do not recognize this as a study limitation because the initial position of the patient’s head cannot be standardized. However, the degree of cervical extension during laryngoscopy is important. The previous literature reported extension at C1/2 produced with ILMAs ranging from 1° to 5° and even as high as 7.4° . Our result is in line with this range. Nevertheless, variability of the results depends on the heterogeneity introduced by methodological and population differences of the studies and experiences of the investigators. The ability to intubate a trachea under glottis visualization with LMA CTrach was reported with higher first attempt success rates compared with LMA Fastrach [14]. Liu et al. found a 98.9% first attempt success rate for LMA CTrach in 100 patients, while Baskett et al. showed a 79.8% success rate for LMA Fastrach with the experience of 500 cases . Bilgin et al. demonstrated a first attempt success rate of intubation of 54% for ILMAs and 90% for C-Trach [13]. Our success rate, which is 75% with LMA CTrach and 55% with LMA Fastrach, is lower than those in the published literature. The diversity of these results is probably due to the methodological differences, the skills of the investigators, and the sample sizes of the mentioned studies. Nevertheless, in the current study, tracheal intubation was defined as successful only if the patient was intubated in two attempts at most with a device and the rate of successful tracheal intubation was 80% with LMA Fastrach, which statistically had the lowest rate.Secondary spinal injury during airway management is not only a result of the mechanical disruption of the unstable segments, but hypoxia is also likely to cause harm . Although both LMA CTrach and LMA Fastrach administrations may prolong intubation, they have established roles in difficult airway management since they do not interrupt ventilation . Obviously, duration of intubation lasts longer with devices that require different maneuvers compared to the laryngoscopes. In the present study, as the most familiar device for anesthetists, the Macintosh laryngoscope intubation was fastest, while the duration of intubation was significantly longer with LMA Fastrach. These findings are in line with those of Bilgin et al., who reported significantly longer mean intubation time with an ILMA compared to C-Trach and McCoy [13]. Nevertheless, none of the patients in our study presented hypoxia throughout the intubation process. Randomizing the patients without considering their Mallampati scores and evaluating airway difficulties might be a limitation of our study. If the selection of the airway device depended on the possibility of a difficult airway, a selection bias would occur for LMA CTrach over blind intubation using LMA Fastrach. Nevertheless, Mallampati scores were identical between the groups. Another possible shortcoming of the study is that it was impossible to blind the investigator to the airway device and only an independent radiologist who measured the angles was unaware of the study group assignments. Several investigators may have had different skill levels and experiences, so only one investigator performed all laryngoscopies in order to minimize any confounding effects. The third limitation is that tracheal intubations were facilitated with muscle relaxants. Sawin et al. [4] suggested that muscle relaxation using neuromuscular blockade might reduce the need for cervical extension during laryngoscopy. However, even in injury settings muscle relaxants are used to ease the insertion of the endotracheal tube. Finally, the study conclusions may be limited since this project examined only two X-ray graphics instead of dynamic fluoroscopy.In conclusion, airway management with minimal neck movement improves the success of the anesthetic management in C-spine surgery. Moreover, there are limited data that may help to understand C-spine kinetics of the patients with cervical pathologies, especially degenerative disorders requiring surgery. It is thus important to be familiar with the different intubation techniques. However, one should also be aware of their effects on cervical extension. We conclude that the reduced C-spine extension during intubation with LMA CTrach makes it a reasonable alternative compared to LMA Fastrach and Macintosh laryngoscopes in cervical surgery where cervical stability is a concern.