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Burrowing organisms augment the availability of important resources for other species. The gopher tortoise (Gopherus polyphemus) is a keystone excavator in open canopy pine-forest ecosystems in the southeastern United States because its burrows are utilized by over 360 species. Across its range, the gopher tortoise is declining, which is thought to negatively affect burrow-associated species and ecosystem functionality. The nine-banded armadillo (Dasypus novemcinctus) is another burrower of similar size that has become syntopically distributed with the gopher tortoise as a result of range expansion. Recent studies have documented vertebrates utilizing armadillo burrows, linking armadillo burrowing to support of local biodiversity similar to the gopher tortoise. We sought to determine the potential for ecological redundancy between gopher tortoises and armadillos and test quantitatively for differences in associate events at their burrows in a mixed-pine–hardwood forest where they co-occur. Using motion activated game cameras to monitor burrows, we compared metrics of vertebrate occurrence between armadillo and tortoise burrows and examined the effects of environmental variables using a series of regression models. A total of 40 vertebrate taxa were observed visiting burrows between October 2019 and December 2020. Richness, diversity, and community composition were not significantly different between the two burrow types. However, associate event counts were significantly greater at tortoise burrows. Burrow and microhabitat variables had varying effects on associate event counts, with consistently positive effects for tortoise burrows, active burrows, and increased richness of tree species, while negative effects were detected for increased canopy cover as well as increased proportions of hardwood trees. Our study provides a framework for testing redundancy of function between syntopic ecosystem engineers, adds to the growing body of work on the ecological significance of armadillo range expansion, and identifies aspects of the habitat that cause fluctuations in the importance of burrows for associate species. [ABSTRACT FROM AUTHOR]

In southern Florida, Gopher Tortoises (Gopherus polyphemus) occupy mesic flatwoods and Florida scrub communities where habitat and climatic conditions differ from other portions of the species' range. Both of these habitats appear suboptimal for tortoises due to saturated soils in mesic flatwoods and low forage abundance in scrub. Nonetheless, these habitats support large numbers of tortoises in southern Florida, albeit at low intensities. We assessed influences of elevation and microtopography on the spatial distributions of tortoise burrows and examined burrow use patterns within six sites at Avon Park Air Force Range in south-central Florida. The six sites differed in dominant soil types and vegetation communities, allowing comparisons of burrow distributions among mesic flatwoods, Florida scrub, and mixed flatwoods-scrub habitats (two replicate sites each). Point-process modeling identified significant influences of microtopography on burrow intensities that superseded the effects of site-wide elevation trends in five of the six sites. The effects of microtopography were most pronounced in flatwoods, suggesting greater reliance on areas of slightly higher elevation in mesic habitat, presumably in response to saturated soils and frequent flooding. Burrow use patterns during an exceedingly wet year also suggested that tortoises respond behaviorally to unsuitable hydrology by moving frequently among burrows that were flooded with groundwater. Microtopographic variation may be an important predictor of small-scale habitat use for fossorial reptiles, especially in mesic soils, which could be readily explored using increasingly available LiDAR-derived elevation data combined with the modeling approach demonstrated here. [ABSTRACT FROM AUTHOR]

Mutations in coiled-coil-helix–coiled-coil-helix domain containing 10 (CHCHD10), a mitochondrial protein of unknown function, cause a disease spectrum with clinical features of motor neuron disease, dementia, myopathy and cardiomyopathy. To investigate the pathogenic mechanisms of CHCHD10, we generated mutant knock-in mice harboring the mouse-equivalent of a disease-associated human S59L mutation, S55L in the endogenous mouse gene. CHCHD10S55L mice develop progressive motor deficits, myopathy, cardiomyopathy and accelerated mortality. Critically, CHCHD10 accumulates in aggregates with its paralog CHCHD2 specifically in affected tissues of CHCHD10S55L mice, leading to aberrant organelle morphology and function. Aggregates induce a potent mitochondrial integrated stress response (mtISR) through mTORC1 activation, with elevation of stress-induced transcription factors, secretion of myokines, upregulated serine and one-carbon metabolism, and downregulation of respiratory chain enzymes. Conversely, CHCHD10 ablation does not induce disease pathology or activate the mtISR, indicating that CHCHD10S55L-dependent disease pathology is not caused by loss-of-function. Overall, CHCHD10S55L mice recapitulate crucial aspects of human disease and reveal a novel toxic gain-of-function mechanism through maladaptive mtISR and metabolic dysregulation. [ABSTRACT FROM AUTHOR]

Abstract: Prohibitin (PHB) is a ubiquitously expressed and evolutionarily conserved mitochondrial protein with multiple functions. We have recently shown that PHB up‐regulation offers robust protection against neuronal injury in models of cerebral ischemia in vitro and in vivo, but the mechanism by which PHB affords neuroprotection remains to be elucidated. Here, we manipulated PHB expression in PC12 neural cells to investigate its impact on mitochondrial function and the mechanisms whereby it protects cells exposed to oxidative stress. PHB over‐expression promoted cell survival, whereas PHB down‐regulation diminished cell viability. Functionally, manipulation of PHB levels did not affect basal mitochondrial respiration, but it increased spare respiratory capacity. Moreover, PHB over‐expression preserved mitochondrial respiratory function of cells exposed to oxidative stress. Preserved respiratory capacity in differentiated PHB over‐expressing cells exposed to oxidative stress was associated with an elongated mitochondrial morphology, whereas PHB down‐regulation enhanced fragmentation. Mitochondrial complex I oxidative degradation was attenuated by PHB over‐expression and increased in PHB knockdown cells. Changes in complex I degradation were associated with alterations of respiratory chain supercomplexes. Furthermore, we showed that PHB directly interacts with cardiolipin and that down‐regulation of PHB results in loss of cardiolipin in mitochondria, which may contribute to destabilizing respiratory chain supercomplexes. Taken together, these data demonstrate that PHB modulates mitochondrial integrity and bioenergetics under oxidative stress, and suggest that the protective effect of PHB is mediated by stabilization of the mitochondrial respiratory machinery and its functional capacity, by the regulation of cardiolipin content. Open Data: Materials are available on https://cos.io/our-services/open-science-badges/ https://osf.io/93n6m/ [ABSTRACT FROM AUTHOR]

The mitochondrial protein prohibitin (PHB) has emerged as an important modulator of neuronal survival in different injury modalities. We previously showed that viral gene transfer of PHB protects CA1 neurons from delayed neurodegeneration following transient forebrain ischemia through mitochondrial mechanisms. However, since PHB is present in all cell types, it is not known if its selective expression in neurons is protective, and if the protection occurs also in acute focal ischemic brain injury, the most common stroke type in humans. Therefore, we generated transgenic mice overexpressing human PHB1 specifically in neurons (PHB1 Tg). PHB1 Tg mice and littermate controls were subjected to transient middle cerebral artery occlusion (MCAo). Infarct volume and sensory-motor impairment were assessed three days later. Under the control of a neuronal promoter (CaMKIIα), PHB1 expression was increased by 50% in the forebrain and hippocampus in PHB1 Tg mice. The brain injury produced by MCAo was reduced by 63 ± 11% in PHB1 Tg mice compared to littermate controls. This reduction was associated with improved sensory-motor performance, suggesting that the salvaged brain remains functional. Approaches to enhance PHB expression may be useful to ameliorate the devastating impact of cerebral ischemia on the brain. [ABSTRACT FROM AUTHOR]

Ten microsatellite loci were characterized for 34 locations from roundtail chub (Gila robusta complex) to better resolve patterns of genetic variation among local populations in the lower Colorado River basin. This group has had a complex taxonomic history and previous molecular analyses failed to identify species diagnostic molecular markers. Our results supported previous molecular studies based on allozymes and DNA sequences, which found that most genetic variance was explained by differences among local populations. Samples from most localities were so divergent species-level diagnostic markers were not found. Some geographic samples were discordant with current taxonomy due to admixture or misidentification; therefore, additional morphological studies are necessary. Differences in spatial genetic structure were consistent with differences in connectivity of stream habitats, with the typically mainstem species, G. robusta, exhibiting greater genetic connectedness within the Gila River drainage. No species exhibited strong isolation by distance over the entire stream network, but the two species typically found in headwaters, G. nigra and G. intermedia, exhibited greater than expected genetic similarity between geographically proximate populations, and usually clustered with individuals from the same geographic location and/or sub-basin. These results highlight the significance of microevolutionary processes and importance of maintaining local populations to maximize evolutionary potential for this complex. Augmentation stocking as a conservation management strategy should only occur under extreme circumstances, and potential source populations should be geographically proximate stocks of the same species, especially for the headwater forms. [ABSTRACT FROM AUTHOR]

The addition of α-deprotonated nitriles to azaheterocycles followed by rearomatization is described. A simple two-step, one-pot procedure for the sequence is also presented. [ABSTRACT FROM AUTHOR]

Abstract: Calcitonin gene-related peptide (CGRP) has been implicated in acute migraine pathogenesis. In an effort to identify novel CGRP receptor antagonists for the treatment of migraine, we have discovered thiazolidinone 49, a potent (K i =30pM, IC50 =1nM), orally bioavailable, CNS-penetrant CGRP antagonist with good pharmacokinetic properties. [Copyright &y& Elsevier]

Landscape features exist at multiple spatial and temporal scales, and these naturally affect spatial genetic structure and our ability to make inferences about gene flow. This article discusses how decisions about sampling of genotypes (including choices about analytical methods and genetic markers) should be driven by the scale of spatial genetic structure, the time frame that landscape features have existed in their current state, and all aspects of a species’ life history. Researchers should use caution when making inferences about gene flow, especially when the spatial extent of the study area is limited. The scale of sampling of the landscape introduces different features that may affect gene flow. Sampling grain should be smaller than the average home-range size or dispersal distance of the study organism and, for raster data, existing research suggests that simplifying the thematic resolution into discrete classes may result in low power to detect effects on gene flow. Therefore, the methods used to characterize the landscape between sampling sites may be a primary determinant for the spatial scale at which analytical results are applicable, and the use of only one sampling scale for a particular statistical method may lead researchers to overlook important factors affecting gene flow. The particular analytical technique used to correlate landscape data and genetic data may also influence results; common landscape-genetic methods may not be suitable for all study systems, particularly when the rate of landscape change is faster than can be resolved by common molecular markers. [ABSTRACT FROM AUTHOR]

Informally gathered species lists are a potential source of data for conservation biology, but most remain unused because of questions of reliability and statistical issues. We applied two alternative analytical methods (contingency tests and occupancy modeling) to a 35-year data set (1973–2007) to test hypotheses about local bird extinction. We compiled data from bird lists collected by expert amateurs and professional scientists in a 2-km2 fragment of lowland tropical forest in coastal Ecuador. We tested the effects of the following on local extinction: trophic level, sociality, foraging specialization, light tolerance, geographical range area, and biogeographic source. First we assessed extinction on the basis of the number of years in which a species was not detected on the site and used contingency tests with each factor to compare the frequency of expected and observed extinction events among different species categories. Then we defined four multiyear periods that reflected different stages of deforestation and isolation of the study site and used occupancy modeling to test extinction hypotheses singly and in combination. Both types of analyses supported the biogeographic source hypothesis and the species-range hypothesis as causes of extinction; however, occupancy modeling indicated the model incorporating all factors except foraging specialization best fit the data. [ABSTRACT FROM AUTHOR]

Radiotelemetry and genetic markers have enabled herpetologists to test the hypothesis that gene flow is restricted among snake dens (a.k.a. "hibernacula"). In the present study, observations of movement and mating were compared to indirect inferences of gene flow based on genetic data (microsatellite DNA loci) for six neighboring hibernacula of the Timber Rattlesnake (Crotalus horridus) in western St. Louis County, Missouri. Behavioral results were consistent with previous findings for this species, with most individuals exhibiting high fidelity to hibernacula, and males exhibiting larger activity areas than females. Behavioral results also supported several long-standing assumptions: males tended to mate at longer distances from hibernacula than females and random pairing between males and females from different hibernacula could not be rejected. Some gravid females moved considerable distances between locations, but most returned to an area near their own hibernaculum to give birth, and birth distances tended to be shorter than the nearest-neighbor distance between hibernacula. Genetic results indicated that the observed number of heterozygotes did not deviate from Hardy-Weinberg expectations at the level of total sample population, but that a significant heterozygote excess was found within hibernacula. A small but significant proportion of the genetic variance in the total population was due to differences among hibernacula; however, Bayesian clustering algorithms could not sort individuals (or groups of hibernacula) into discrete demes. Results suggest that patterns of genetic differentiation observed among hibernacula of the Timber Rattlesnake are better explained by behavioral and demographic factors resulting in natal philopatry, sex-biased dispersal, and/or a limited number of breeding adults, than by the balance of gene flow and drift, per se. [ABSTRACT FROM AUTHOR]

Populations of the Desert Massasauga Rattlesnake (Sistrurus catenatus edwardsii) have declined rapidly as desert grassland communities have become reduced and fragmented. To provide information useful for management of remaining populations, the genetic characteristics (based on microsatellite DNA loci) of the last demonstrably extant population in the state of Arizona were compared to a population in the Rio Grande Valley of New Mexico. Results indicated that genetic diversity was relatively high in both populations, with statistically significant heterozygote deficiencies detected at only one of six loci in each population. Contingency tests, Wright's F-statistics, and Bayesian clustering algorithms all indicated substantial subdivision between populations in Arizona and New Mexico, but only contingency tests supported differentiation within the Arizona population. A preliminary hierarchical analysis of variance (incorporating both our data and published microsatellite data for the Eastern Massasauga) indicated that 73% of the total molecular variance was explained by variation within populations, with variation between the two subspecies accounting for 15% of the total variance. Results support the high conservation value of individual populations, as well as the need for further population genetic studies of the Desert Massasauga Rattlesnake. [ABSTRACT FROM AUTHOR]

The mechanism(s) underlying the beneficial effects of adult mesenchymal stem cells (MSCs) after myocardial infarction (MI) is poorly understood. One possible explanation is the ability of MSCs to secrete cytokines, which modulate cardiomyocyte survival and function. MSCs express at least two cytoprotective cytokines, hepatocyte growth factor (HGF) and stromal cell-derived factor-1 α (CXCL12). The aim of our study was to compare the effects of these two cytokines administered acutely post-MI. We subjected adult male Lewis rats to myocardial ischemia/reperfusion injury. Immediately upon reperfusion, polymers saturated with HGF or CXCL12 were placed onto the infarcted anterior wall and the rats were allowed to recover. Echocardiographic analysis at 4 wk post-MI to assess left ventricular (LV) function revealed that LV ejection fraction was increased in the HGF treated group compared with the phosphate-buffered saline (PBS) control group. Likewise, LV end diastolic dimension was reduced in the HGF treated group compared with the PBS control group. Similarly, invasive hemodynamics at 12 wk showed improved contractility and relaxation in the HGF treated group compared with the PBS control group. In contrast, no significant effect on LV function was seen in the CXCL12 treated group. To determine the potential mechanism for this effect, infarct size (IFS) at 72 h was determined. IFS was decreased 4.2-fold in the HGF treated group compared with the PBS control group. Thus, HGF acutely post-MI using polymer delivery reduces IFS, leading to beneficial effects on post-MI LV remodeling. [Copyright &y& Elsevier]

The article focuses on the study conducted regarding the effectiveness of the combining elastic and free weight resistance in providing power and strengths in the athletes. The study, administered to 44 young resistance-trained players, aims to determine the effectivity of this resistance compared to free weight resistance (FWR) alone. The result indicates that combined training is more effective in developing the lower and upper body strength compared to FWR.

I tested six microsatellite DNA primer pairs developed for the massasauga rattlesnake ( Sistrurus catenatus) on a sample population of the timber rattlesnake ( Crotalus horridus). It had been speculated in a previous publication that cross-species amplification would not be worthwhile across the two rattlesnake genera. However, for this primer set (the only one currently published for the genus Sistrurus), successful amplification at each locus was accomplished for all loci with an annealing temperature of 57 °C and locus-specific buffer conditions. Each locus was polymorphic, with the number of alleles per locus ranging from two to 12. Significant heterozygote deficits were detected for three loci (Scu01, Scu05 and Scu07 ). For Scu01, all individuals were homozygous for the same allele except one female who was homozygous for a different allele. This same female was also homozygous for a rare allele at Scu07. When this female was removed from the data set, the number of observed heterozygotes at Scu01 and Scu07 did not differ significantly from random expectations. However, a large heterozygote deficit persisted at Scu05 (despite subsampling), suggesting that this locus may not be useful for population genetic studies of timber rattlesnakes. Despite some limitations, this set of primers may be a useful complement to those already developed for the genus Crotalus. Moreover, the results of this study seem to provide new justification for further studies of cross-species amplification of microsatellite loci across the two rattlesnake genera. [ABSTRACT FROM AUTHOR]

KCNH2 (hERG1) encodes the α-subunit proteins for the rapidly activating delayed rectifier K+ current (IKr), a major K+ current for cardiac myocyte repolarization. In isolated myocytes IKr frequently is small in amplitude or absent, yet KCNH2 channels and IKr are targets for drug block or mutations to cause long QT syndrome. We hypothesized that KCNH2 channels and IKr are uniquely sensitive to enzymatic damage. To test this hypothesis, we studied heterologously expressed K+, Na+, and L-type Ca2+ channels, and in ventricular myoctyes IKr, slowly activating delayed rectifier K+ current (IKs), and inward rectifier K+ current (IKl), by using electrophysiological and biochemical methods. 1) Specific exogenous serine proteases (protease XIV, XXIV, or proteinase K) selectively degraded KCNH2 current (IKCNH2) and its mature channel protein without damaging cell integrity and with minimal effects on the other channel currents; 2) immature KCNH2 channel protein remained intact; 3) smaller molecular mass KCNH2 degradation products appeared; 4) protease XXIV selectively abolished IKr; and 5) reculturing HEK-293 cells after protease exposure resulted in the gradual recovery of IKCN2 and its mature channel protein over several hours. Thus the channel protein for IKCNH2 and IKr is uniquely sensitive to proteolysis. Analysis of the degradation products suggests selective proteolysis within the S5-pore extracellular linker, which is structurally unique among Kv channels. These data provide 1) a new mechanism to account for low IKr density in some isolated myocytes, 2) evidence that most complexly glycosylated KCNH2 channel protein is in the plasma membrane, and 3) new insight into the rate of biogenesis of KCNH2 channel protein within cells. [ABSTRACT FROM AUTHOR]

Mutations in the human ether-a-go-go-related gene (HERG) cause long QT syndrome. We previously showed that the HERG N470D mutation expressed as homotetrameric channels causes a protein trafficking defect, and this can be corrected by the HERG channel blocking drug E-4031. The N470D mutant also has been reported to cause dominant negative suppression of HERG current when coexpressed with wild-type channel subunits. The aims of this study were 1) to investigate the molecular mechanism responsible for the dominant negative effect of the N470D mutant co-expressed with wild-type subunits and 2) to test whether the trafficking defective heteromeric channels could be pharmacologically rescued by E-4031. Using a combination of immunoprecipitation and Western blot methods, we showed that N470D mutant and wild-type HERG subunits were physically associated in the endoplasmic reticulum as heteromeric channels. The coassembly resulted in the retention of both wild-type and N470D subunits in the endoplasmic reticulum. Culturing cells in E-4031 increased the cell surface expression of these channels, although with an altered electrophysiological phenotype. These results suggest that the dominant negative effect of the N470D wild-type coassembled channels is caused by retention of heteromeric channels in the endoplasmic reticulum and that the trafficking defect of these channels can be corrected by specific pharmacological strategies. [ABSTRACT FROM AUTHOR]

Several mutations in the human ether-a-go-go-related K[sup +] channel gene (HERG or KCNH2) cause long QT syndrome (LQT2) by reducing the intracellular transport (trafficking) of the channel protein to the cell surface. Drugs that bind to and block HERG channels (i.e. E4031) rescue the surface expression of some trafficking defective LQT2 mutations. Because these drugs potently block HERG current, their ability to correct congenital LQT is confounded by their risk of causing acquired LQT. We tested the hypothesis that pharmacological rescue can occur without HERG channel block. Thapsigargin (1 µM), a sarcoplasmic/endoplasmic reticulum Ca[sup 2+]ATPase inhibitor, rescued the surface expression of G601S, and it did so without blocking current. Thapsigargin-induced rescue and E4031-induced rescue caused complex glycosylation that was evident within 3 h of drug exposure. Disruption of the Golgi apparatus with brefeldin A prevented thapsigargin- and E4031-induced rescue of I[sub G01S]. Confocal imaging showed that G601S protein is predominantly "trapped" intracellularly and that both thapsigargin and E4031 promote its relocation to the surface membrane. We also studied two other trafficking defective LQT2 mutations. Thapsigargin rescued the C terminus mutation F805C but not N470D, whereas E4031 rescued N470D but not F805C. Other sarcoplasmic/endoplasmic reticulum Ca[sup 2+]ATPase inhibitors did not rescue G601S or F805C. This study 1) supports the hypothesis that the LQT2 trafficking defective phenotype can be reversed without blocklng the channel; 2) demonstrates pharmacological rescue of a C terminus LQT2 mutation; and 3) shows that thapsigargin can correct trafficking defective phenotypes in more than one channel type and disease (i.e. LQT2 and cystic fibrosis). [ABSTRACT FROM AUTHOR]

Presents a study that used the approaches of site-directed mutagenesis and biochemical modification to inhibit asparagine (N)-linked glycosylation and examined the role of glycosylation in the cell surface expression and turnover of human ether- à -go-go-related gene (HERG). Information on HERG; Details of the inhibition of N-linked glycosylation by tunicamycin; Cell surface expression of wild-type HERG and N-linked glycosylation.

A two-step reaction sequence yielding 2-aminooxazolines under mild conditions was developed. Both electron-rich and electron-deficient styrenes as well as functionalized primary and secondary amides could be used in the reaction to afford 2-aminooxazolines in reasonable yields. [ABSTRACT FROM AUTHOR]

(J Card Surg 2011;26:63-65) [ABSTRACT FROM AUTHOR]

Prohibitin (PHB) is a critical protein involved in many cellular activities. In brain, PHB resides in mitochondria, where it forms a large protein complex with PHB2 in the inner TFmembrane, which serves as a scaffolding platform for proteins involved in mitochondrial structural and functional integrity. PHB overexpression at moderate levels provides neuroprotection in experimental brain injury models. In addition, PHB expression is involved in ischemic preconditioning, as its expression is enhanced in preconditioning paradigms. However, the mechanisms of PHB functional regulation are still unknown. Observations that nitric oxide (NO) plays a key role in ischemia preconditioning compelled us to postulate that the neuroprotective effect of PHB could be regulated by NO. Here, we test this hypothesis in a neuronal model of ischemia-reperfusion injury and show that NO and PHB are mutually required for neuronal resilience against oxygen and glucose deprivation stress. Further, we demonstrate that NO post-translationally modifies PHB through protein S-nitrosylation and regulates PHB neuroprotective function, in a nitric oxide synthase-dependent manner. These results uncover the mechanisms of a previously unrecognized form of molecular regulation of PHB that underlies its neuroprotective function. [ABSTRACT FROM AUTHOR]

Gunnison's prairie dogs, Cynomys gunnisoni , frequently utter a repetitive series (a 'bout') of alarm calls (called 'barks') in response to a variety of terrestrial predators. Previous work has suggested that the structure of barks is finely tuned in ways that provide highly specific information about the identity of the predator. We re-evaluated these findings by presenting individually marked adult females with taxidermy mounts of three terrestrial predators of prairie dogs: a coyote, Canis latrans , a bobcat, Lynx rufus , and an American badger, Taxidea taxus , as well as one control stimulus (a cardboard box). Significant variation among individuals was evident from analyses of 10 measures associated with the general pattern of barking (e.g. latency to begin barking, bout duration, average number of barks/bout, overall bark rate, interbout interval) and also from eight structural features of individual barks (e.g. frequency and duration measures of the fundamental frequency band). When controlling for the identity of alarm callers, we found little evidence for differences in these same variables in response to different stimuli. We conclude that, like several other species of ground-dwelling squirrels, Gunnison's prairie dogs have an individually distinctive alarm call that does not differ for different predators. Our results conflict with earlier claims of extreme specificity in the alarm calls of Gunnison's prairie dogs. Highlights • We tested whether Gunnison's prairie dog alarm calls show referential specificity. • Controlling for caller ID, we found little evidence of call variation to four stimuli. • However, individuals' alarm calls were distinct from one other. • Previous studies not considering individual variation need to be re-evaluated. [ABSTRACT FROM AUTHOR]

A major technology break in snake biology was the publication of surgical protocols for implanting radiotransmitters in the body cavities of snakes. While many researchers have reported using some variant of these protocols successfully, protocol details often vary from study to study and best-practice procedures are not easily determinable given the variety of circumstances over which studies occur. Although professional society standards and federal regulations make explicit recommendations about this surgical procedure, some of the nonstandard techniques recommended for this protocol may raise the eyebrows of institutional animal care and use committees. In this commentary we discuss regulatory and logistical aspects of the intracoelmic radiotransmitter implant procedure for snakes, and we provide a pragmatic framework for choosing among surgical variables. [ABSTRACT FROM AUTHOR]

The article presents chemical equations related to the article "Addition of α-Lithiated Nitriles to Azaheterocycles" by C. Anderson, J. Moreno and S. Hadida, published in the periodical "Synlett" in 2014.

The electrocardiogram (ECG) empowered clinician scientists to measure the electrical activity of the heart noninvasively to identify arrhythmias and heart disease. Shortly after the standardization of the 12-lead ECG for the diagnosis of heart disease, several families with autosomal recessive (Jervell and Lange-Nielsen Syndrome) and dominant (Romano–Ward Syndrome) forms of long QT syndrome (LQTS) were identified. An abnormally long heart rate-corrected QT-interval was established as a biomarker for the risk of sudden cardiac death. Since then, the International LQTS Registry was established; a phenotypic scoring system to identify LQTS patients was developed; the major genes that associate with typical forms of LQTS were identified; and guidelines for the successful management of patients advanced. In this review, we discuss the molecular and cellular mechanisms for LQTS associated with missense variants in KCNQ1 (LQT1) and KCNH2 (LQT2). We move beyond the "benign" to a "pathogenic" binary classification scheme for different KCNQ1 and KCNH2 missense variants and discuss gene- and mutation-specific differences in K+ channel dysfunction, which can predispose people to distinct clinical phenotypes (e.g., concealed, pleiotropic, severe, etc.). We conclude by discussing the emerging computational structural modeling strategies that will distinguish between dysfunctional subtypes of KCNQ1 and KCNH2 variants, with the goal of realizing a layered precision medicine approach focused on individuals. [ABSTRACT FROM AUTHOR]

The nine-banded armadillo (Dasypus novemcinctus) is unique among mammals because females produce litters of genetically identical quadruplets via monozygotic polyembryony. This unusual form of reproduction could have profound impacts on the spatial genetic structure of populations of armadillos, but at present it is unclear whether littermates remain together as adults, or if sex-biased dispersal occurs. The goal of our study was to determine whether fine-scale spatial genetic structure (FSGS) within a population of armadillos diminishes with age (i.e., from juvenile to adult due to the dispersal of littermates away from one another), and if the degree of FSGS for a given age class differs between the sexes. We obtained genotype data at seven microsatellite DNA loci for 421 individuals in a wild population of armadillos inhabiting the Yazoo National Wildlife Refuge in western Mississippi. Correlogram analyses based on measures of spatial genetic autocorrelation showed weak but significant FSGS that was driven by positive spatial genetic autocorrelation among both male and female juveniles and adult males, but not adult females. Positive spatial genetic autocorrelation among adult males could be due to either female-biased dispersal or high variance in male reproductive success. Further work is required to discriminate between these two possibilities. [ABSTRACT FROM AUTHOR]

The objective of this study was to optimize the methodology for spectrophotometric determination of sperm concentration in Atlantic croaker Micropogonias undulatus L. milt and to estimate its potential for short-term cold-storage. The spectrophotometric determination of sperm concentration was evaluated using milt samples from six males serially diluted in Hank's balanced salt solution at 200 mOsm kg−1 (HBSS). The predictive power of regression models between sperm concentration and absorbance was determined from 200 to 500 nm and found to be highest within the visible spectrum despite a peak of milt absorbance at 288 nm. Absorbance reading at 400 nm was selected for further analysis to maximize the absorbance of the sample hence the sensitivity of the method while minimizing the impact of potential sample contamination with blood. The standard-curve of correlation between sperm absorbance at 400 nm and concentration was validated and held an accuracy ranging between −7.40% and +4.56% across males. Total sperm motility duration and the proportion of motile spermatozoa were significantly higher in milt samples diluted 1:3 in HBSS than in the undiluted control during up to 30 h of cold-storage.The methodologies investigated in this study can be applied to optimize sperm usage and achieve predictable artificial fertilization protocols in Atlantic croaker. [ABSTRACT FROM AUTHOR]

KCNH2 encodes Kv11.1 and underlies the rapidly activating delayed rectifier K+ current (IKr) in the heart. Loss-of-function KCNH2 mutations cause the type 2 long QT syndrome (LQT2), and most LQT2-linked missense mutations inhibit the trafficking of Kv11.1 channels. Drugs that bind to Kv11.1 and block IKr (e.g., E-4031) can act as pharmacological chaperones to increase the trafficking and functional expression for most LQT2 channels (pharmacological correction). We previously showed that LQT2 channels are selectively stored in a microtubuledependent compartment within the endoplasmic reticulum (ER). We tested the hypothesis that pharmacological correction promotes the trafficking of LQT2 channels stored in this compartment. Confocal analyses of cells expressing the trafficking-deficient LQT2 channel G601S showed that the microtubule-dependent ER compartment is the transitional ER. Experiments with E-4031 and the protein synthesis inhibitor cycloheximide suggested that pharmacological correction promotes the trafficking of G601S stored in this compartment. Treating cells in E-4031 or ranolazine (a drug that blocks IKr and has a short half-life) for 30 min was sufficient to cause pharmacological correction. Moreover, the increased functional expression of G601S persisted 4-5 h after drug washout. Coexpression studies with a dominant- negative form of Rab11B, a small GTPase that regulates Kv11.1 trafficking, prevented the pharmacological correction of G601S trafficking from the transitional ER. These data suggest that pharmacological correction quickly increases the trafficking of LQT2 channels stored in the transitional ER via a Rab11B-dependent pathway, and we conclude that the pharmacological chaperone activity of drugs like ranolazine might have therapeutic potential. [ABSTRACT FROM AUTHOR]

Long QT syndrome (LQTS) is a cardiac repolarization disorder that can lead to arrhythmias and sudden death. Chromosome 7-1inked inherited LQTS (LQT2) is caused by mutations in human ether-a-go-go-related gene (HERG; KCNH2), whereas drug-induced LQTS is caused primarily by HERG channel block. Many common polymorphisms are functionally silent and have been traditionally regarded as benign and without physiological consequence. However, the identification of common nonsynonymous single nucleotide polymorphisms (nSNPs; i.e., amino-acid coding variants) with functional phenotypes in the SCN5A Na+ channel and MiRP1 K+ channel β-subunit have challenged this viewpoint. In this report, we test the hypothesis that common missense HERG polymorphisms alter channel physiology. Comprehensive mutational analysis of HERG was performed on genomic DNA derived from a population-based cohort of sudden infant death syndrome and two reference allele cohorts derived from 100 African American and 100 Caucasian individuals. Amino acidencoding variants were considered common polymorphisms if they were present in at least two of the three study cohorts with an allelic frequency >0.5%. Four nSNPs were identified: K897T, P967L, R1047L, and Q1068R. Wild-type (WT) and polymorphic channels were heterologously expressed in human embryonic kidney cells, and biochemical and voltage-clamp techniques were used to characterize their functional properties. All channel types were processed similarly, but several electrophysiological differences were identified: 1) K897T current density was lower than the other polymorphic channels; 2) K897T channels activated at more negative potentials than WT and R1047L; 3) K897T and Q1068R channels inactivated and recovered from inactivation faster than WT, P967L, and R1047L channels; and 4) K897T channels showed subtle differences compared with WT channels when stimulated with an action potential waveform. In contrast to K897T and Q1068R channels, P967L and R1047L channels were electrophysiologically indistinguishable from WT channels. All HERG channels had similar sensitivity to block by cisapride. Therefore, some HERG polymorphic channels are electrophysiologically different from WT channels. [ABSTRACT FROM AUTHOR]

Significant advances in our understanding of the molecular mechanisms that cause congenital long QT syndrome (LQTS) have been made. A wide variety of experimental approaches, including heterologous expression of mutant ion channel proteins and the use of inducible pluripotent stem cell-derived cardiomyocytes (iPSC-CMs) from LQTS patients offer insights into etiology and new therapeutic strategies. This review briefly discusses the major molecular mechanisms underlying LQTS type 2 (LQT2), which is caused by loss-of-function (LOF) mutations in the KCNH2 gene (also known as the human ether-à-go-go-related gene or hERG). Almost half of suspected LQT2-causing mutations are missense mutations, and functional studies suggest that about 90% of these mutations disrupt the intracellular transport, or trafficking, of the KCNH2-encoded Kv11.1 channel protein to the cell surface membrane. In this review, we discuss emerging strategies that improve the trafficking and functional expression of trafficking-deficient LQT2 Kv11.1 channel proteins to the cell surface membrane and how new insights into the structure of the Kv11.1 channel protein will lead to computational approaches that identify which KCNH2 missense variants confer a high-risk for LQT2. [ABSTRACT FROM AUTHOR]

Introduction: Inherited long QT syndrome type 2 (LQT2) results from loss-of-function mutations in the KCNH2 gene encoding the hERG channel, which conducts I Kr, the rapid component of the delayed rectifier K+ current. The N-terminal Per-Arnt-Sim (PAS) domain present on the hERG1a subunit, but not the hERG1b subunit, is the site of multiple LQT2-linked missense variants. The mechanism of loss of function by many of these missense PAS variants is unclear given conflicting results from different heterologous expression systems expressing hERG1a. Hypothesis: Patient-specific LQT2 human iPSC-cardiomyocytes (hiPSC-CMs) which naturally express hERG1a/1b carrying the KCNH2 H70R variant in the PAS domain will exhibit loss of I Kr associated with APD prolongation due to impaired channel protein trafficking. Methods and Results: Human iPSCs were derived from a patient carrying the LQT2-associated PAS domain mutation KCNH2 H70R, which has been reported to cause impaired hERG channel trafficking without effects on channel gating when expressed in HEK 293 cells but accelerated deactivation kinetics of IhERG when expressed in Xenopus laevis oocytes. Two clones of KCNH2 H70R and unrelated control hiPSCs (DF19-9-11T) were differentiated using monolayer-base, small molecule protocol to CMs, evaluated with whole-cell patch clamp. Action potentials from single hiPSC-CMs paced at 1Hz were prolonged in the hERG-H70R group compared to control (APD90 439.9 ± 15.3 ms vs. 363.7 ± 29.0ms, p =0.003, H70R: n=11, control: n=9, Temp 36 ± 1°C). Voltage clamp studies showed hERG-H70R hiPSC-CMs had a significantly smaller peak tail IKr current density (1.1 ± 0.3 vs. 2.9 ± 0.5 pA/pF, p <0.001, H70R: n=11, control: n=7, Temp 36 ± 1°C). The voltage dependence of IKr activation (V½ and k) were not affected by the mutation; however, the fast (τf) and slow (τs) deactivation time constants were significantly decreased in hERG-H70R hiPSC-CMs. Further, Western blot characterization revealed impaired trafficking of hERG-H70R channels relative to control. Conclusions: The LQT2 PAS domain variant hERG-H70R results in loss of function of IKr by both reduced membrane trafficking and accelerated deactivation of hERG in a hiPSC-CMs model which informs therapeutic approaches. [ABSTRACT FROM AUTHOR]