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We explore the effect of including progenitor mass- and metallicity-dependent yields, supernova rates and energetics on variations in elemental abundance ratios (particularly [α/Fe]) in dwarf galaxies. To understand how the scatter and overall trends in [α/Fe] are affected by including variable metal yields from a discretely sampled initial mass function, we run FIRE simulations of a dwarf galaxy (M★(z = 0) ∼ 106 M☉) using nucleosynthetic yields from the NuGrid data base that depend on the stellar progenitor mass and metallicity. While NuGrid exhibits lower aggregate α-element production than default FIRE yields, we find that its explicit mass dependence, even when including turbulent metal diffusion, substantially widens the intrinsic scatter in the simulated [Fe/H]-[α/Fe] – a phenomenon visible in some observations of dwarf galaxies.

In recent years, venture capital approaches have delivered impressive results in identifying and funding promising health discoveries and bringing them to market. This success has inspired public sector experiments with "social venture capital" approaches to address the dearth of affordable treatment and prevention for diseases of the developing world. Employing the same focus on well-defined and measurable objectives, and the same type of connections to pool and deploy resources as their for-profit counterparts, social venture capitalists seek to use the tools and incentives of capitalism to solve one of its biggest failures: the lack of drugs and vaccines for diseases endemic to low-income populations. As part of a larger trend of partnerships emerging in health product donation and distribution, public-private partnerships for pharmaceutical development have led research and development (R & D) efforts to generate more accessible and efficacious products for diseases such as malaria, tuberculosis, and AIDS. In this article, three R & D-focused partnerships are explored: the International AIDS Vaccine Initiative; the Medicines for Malaria Venture; and the newly formed Global Alliance for TB Drug Development. The article highlights key elements essential to the success of these ventures.

Dystocia, or obstructed labor, is a well-documented phenomenon in various captive vertebrates, including fish. However, despite the documentation of dystocia in several viviparous (live-bearing) Chondrichthyan species (i.e., sharks, rays, skates, and chimaeras), there are no reports to date of dystocia in any oviparous (egg-laying) species. Here we present a case of a captive female epaulette shark (Hemiscyllium ocellatum) that demonstrated symptoms of dystocia in a research-related captive breeding programme. This communication serves as documentation that dystocia can occur in oviparous Chondrichthyans, and this information can help inform researchers and veterinary practitioners for improved care., (© 2024 The Author(s). Journal of Fish Biology published by John Wiley & Sons Ltd on behalf of Fisheries Society of the British Isles.)

(1) Background: To identify reasons for the persistence of surgical ligation of the patent ductus arteriosus (PDA) in premature infants after the 2019 Food and Drug Administration (FDA) approval of transcatheter device closure; (2) Methods: We performed a 10-year (2014-2023) single-institution retrospective study of premature infants (<37 weeks) and compared clinical characteristics and neonatal morbidities between neonates that underwent surgical ligation before (epoch 1) and after (epoch 2) FDA approval of transcatheter closure; (3) Results: We identified 120 premature infants that underwent surgical ligation ( n = 94 before, n = 26 after FDA approval). Unfavorable PDA morphology, active infection, and recent abdominal pathology were the most common reasons for surgical ligation over device occlusion in epoch 2. There were no differences in demographics, age at closure, or outcomes between infants who received surgical ligation in the two epochs; (4) Conclusions: Despite increasing trends for transcatheter PDA closure in premature infants, surgical ligation persists due to unfavorable ductal morphology, active infection, or abdominal pathology.

Rising ocean temperatures are threatening marine species and populations worldwide, and ectothermic taxa are particularly vulnerable. Echinoderms are an ecologically important phylum of marine ectotherms and shifts in their population dynamics can have profound impacts on the marine environment. The effects of warming on echinoderms are highly variable across controlled laboratory-based studies. Accordingly, synthesis of these studies will facilitate the better understanding of broad patterns in responses of echinoderms to ocean warming. Herein, a meta-analysis incorporating the results of 85 studies (710 individual responses) is presented, exploring the effects of warming on various performance predictors. The mean responses of echinoderms to all magnitudes of warming were compared across multiple biological responses, ontogenetic life stages, taxonomic classes, and regions, facilitated by multivariate linear mixed effects models. Further models were conducted, which only incorporated responses to warming greater than the projected end-of-century mean annual temperatures at the collection sites. This meta-analysis provides evidence that ocean warming will generally accelerate metabolic rate (+32%) and reduce survival (-35%) in echinoderms, and echinoderms from subtropical (-9%) and tropical (-8%) regions will be the most vulnerable. The relatively high vulnerability of echinoderm larvae to warming (-20%) indicates that this life stage may be a significant developmental bottleneck in the near-future, likely reducing successful recruitment into populations. Furthermore, asteroids appear to be the class of echinoderms that are most negatively affected by elevated temperature (-30%). When considering only responses to magnitudes of warming representative of end-of-century climate change projections, the negative impacts on asteroids, tropical species and juveniles were exacerbated (-51%, -34% and -40% respectively). The results of these analyses enable better predictions of how keystone and invasive echinoderm species may perform in a warmer ocean, and the possible consequences for populations, communities and ecosystems., Competing Interests: The authors declare no competing interests., (© 2023 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Body condition is an important proxy for the overall health and energetic status of fishes. The classically used Fulton's condition factor requires length and mass measurements, but mass can be difficult to obtain in large species. Girth measurements can replace mass for wild pelagic sharks. However, girth-calculated condition has not been validated against Fulton's condition factor intraspecifically, across ontogeny or reproduction, or in a controlled setting. We used the epaulette shark (Hemiscyllium ocellatum), because they are amenable to captive reproduction, to track fine-scale body condition changes across life stages, oviparous reproduction and between condition indices. We measured four girths, total length and mass of 16 captive epaulette sharks across 1 year and tracked female reproduction daily. We also collected length and mass data from an additional 72 wild-caught sharks and 155 sharks from five previous studies and two public aquaria to examine the relationship between length and mass for this species. Even though data were derived from a variety of sources, a predictable length-mass relationship (R 2  = 0.990) was achievable, indicating that combining data from a variety of sources could help overcome knowledge gaps regarding basic life history characteristics. We also found that condition factor decreased during early life stages, then increased again into adulthood, with predictable changes across the female reproductive cycle. Finally, we determined that both Fulton's and girth condition analyses were comparable. Outcomes from this study uniquely provide body condition changes across the complete life history, including fine-scale female reproductive stages, and validate the use of girths as a nonlethal whole-organism energetic assessment for fishes., (© 2023 The Authors. Journal of Fish Biology published by John Wiley & Sons Ltd on behalf of Fisheries Society of the British Isles.)

The utilization of extracorporeal membrane oxygenation (ECMO) for cardiopulmonary support continues to increase globally, with > 190,000 ECMO cases reported to the international Extracorporeal Life Support Organization Registry. The present review aims to synthesize important contributions to the literature surrounding the management of mechanical ventilation, prone positioning, anticoagulation, bleeding complications, and neurologic outcomes for infants, children, and adults undergoing ECMO in 2022. Additionally, issues related to cardiac ECMO, Harlequin syndrome, and anticoagulation during ECMO will be discussed., Competing Interests: Dr Wheeler discloses a relationship with Bunnell. Mr Bullock discloses no conflicts of interest., (Copyright © 2023 by Daedalus Enterprises.)

Owing to climate change, most notably the increasing frequency of marine heatwaves and long-term ocean warming, better elucidating the upper thermal limits of marine fishes is important for predicting the future of species and populations. The critical thermal maximum (CT max ), or the highest temperature a species can tolerate, is a physiological metric that is used to establish upper thermal limits. Among marine organisms, this metric is commonly assessed in bony fishes but less so in other taxonomic groups, such as elasmobranchs (subclass of sharks, rays and skates), where only thermal acclimation effects on CT max have been assessed. Herein, we tested whether three life history stages, sex and body size affected CT max in a tropical elasmobranch, the epaulette shark ( Hemiscyllium ocellatum ), collected from the reef flats surrounding Heron Island, Australia. Overall, we found no difference in CT max between life history stages, sexes or across a range of body sizes. Findings from this research suggest that the energetically costly processes (i.e. growth, maturation and reproduction) associated with the life history stages occupying these tropical reef flats do not change overall acute thermal tolerance. However, it is important to note that neither embryos developing in ovo , neonates, nor females actively encapsulating egg cases were observed in or collected from the reef flats. Overall, our findings provide the first evidence in an elasmobranch that upper thermal tolerance is not impacted by life history stage or size. This information will help to improve our understanding of how anthropogenic climate change may (or may not) disproportionally affect particular life stages and, as such, where additional conservation and management actions may be required., (© The Author(s) 2022. Published by Oxford University Press and the Society for Experimental Biology.)

The ductus arteriosus (DA) has a paradoxical biological role in neonates with congenital heart disease (CHD) and can present with significant management challenges. Critical congenital cardiac lesions rely on the patency of the DA to provide either systemic or pulmonary blood flow. A patent DA (PDA) that remains open can also have adverse consequences depending on the degree of systemic to-pulmonary shunting and volume of ductal steal. As such, the presence of a PDA may pose a challenge in the medical management and timing of surgical repair. In this perspective article, we provide an understanding of the role of the DA in the circulatory system in neonates with CHD and discuss traditional and emerging approaches to support the pulmonary and systemic circulations with manipulation of the DA., (© 2022. The Author(s), under exclusive licence to Springer Nature America, Inc.)

Biological rhythms that are mediated by exogenous factors, such as light and temperature, drive the physiology of organisms and affect processes ranging from cellular to population levels. For elasmobranchs (i.e. sharks, rays, and skates), studies documenting diel activity and movement patterns indicate that many species are crepuscular or nocturnal in nature. However, few studies have investigated the rhythmicity of elasmobranch physiology to understand the mechanisms underpinning these distinct patterns. Here, we assess diel patterns of metabolic rates in a small meso-predator, the epaulette shark ( Hemiscyllium ocellatum ), across ecologically relevant temperatures and upon acutely removing photoperiod cues. This species possibly demonstrates behavioral sleep during daytime hours, which is supported herein by low metabolic rates during the day and a 1.7-fold increase in metabolic rates at night. From spring to summer seasons, where average average water temperature temperatures for this species range 24.5 to 28.5 °C, time of day, and not temperature, had the strongest influence on metabolic rate. These results indicate that this species, and perhaps other similar species from tropical and coastal environments, may have physiological mechanisms in place to maintain metabolic rate on a seasonal time scale regardless of temperature fluctuations that are relevant to their native habitats.

Objective: To identify risk factors associated with high-frequency ventilation (HFV) following definitive closure of the patent ductus arteriosus (PDA)., Methods: We performed a retrospective study of premature infants (<37 weeks) who were mechanically ventilated before and after surgical or transcatheter PDA closure. Primary outcome was HFV requirement within 24 h of procedure. Logistic regression was used to estimate clinical associations with post procedure HFV requirement., Results: We identified 110 infants who were mechanically ventilated before PDA closure, of which 48 (44%) escalated to HFV within 24 h after closure. In the multivariable model, surgical ligation (OR 21.5, 95% CI 1.6-284), elevated Respiratory Severity Score (RSS) 1 h post-procedure (OR 1.78, 95% CI 1.07-2.99) and 12 h post-procedure (OR 2.12, 95% CI 1.37-3.26) were independent predictors of HFV., Conclusion: Surgical ligation and elevated RSS values over the first 12 h after PDA closure are risk factors for HFV., (© 2021. The Author(s), under exclusive licence to Springer Nature America, Inc.)

A persistent patent ductus arteriosus (PDA) can have significant clinical consequences in preterm infants, depending on the degree of left-to-right shunting, its impact on cardiac performance, and associated perinatal risk factors that can mitigate or exacerbate the shunt. Although the best management strategy remains contentious, PDAs that have contraindications to, or have failed medical management have historically undergone surgical ligation. Recently smaller occluder devices and delivery systems have allowed for minimally invasive closure in the catheterization laboratory even in extremely premature infants. The present review summarizes the pathophysiologic manifestations, treatment options and management of hemodynamically significant PDA in preterm infants. Additionally, we review the available literature surrounding the respiratory support and outcomes of preterm infants following definitive PDA closure., (Copyright © 2022 by Daedalus Enterprises.)

Climate change is affecting thermal regimes globally, and organisms relying on their environment to regulate biological processes face unknown consequences. In ectotherms, temperature affects development rates, body condition, and performance. Embryonic stages may be the most vulnerable life history stages, especially for oviparous species already living at the warm edge of their distribution, as embryos cannot relocate during this developmental window. We reared 27 epaulette shark (Hemiscyllium ocellatum) embryos under average summer conditions (27 °C) or temperatures predicted for the middle and end of the twenty-first century with climate change (i.e., 29 and 31 °C) and tracked growth, development, and metabolic costs both in ovo and upon hatch. Rearing sharks at 31 °C impacted embryonic growth, yolk consumption, and metabolic rates. Upon hatch, 31 °C-reared sharks weighed significantly less than their 27 °C-reared counterparts and exhibited reduced metabolic performance. Many important growth and development traits in this species may peak after 27 °C and start to become negatively impacted nearing 31 °C. We hypothesize that 31 °C approximates the pejus temperature (i.e., temperatures at which performance of a trait begin to decline) for this species, which is alarming, given that this temperature range is well within ocean warming scenarios predicted for this species' distribution over the next century.

Background: High-frequency jet ventilation (HFJV) has been used in conjunction with conventional ventilation for premature infants with respiratory failure. We sought to identify parameters that were associated with mortality in subjects who underwent HFJV., Methods: Subjects were enrolled if birthweight was ≤ 2,000 g and they were ≤ 34 weeks gestational age. Subjects were excluded if they received HFJV at the time of admission because we aimed to study subjects who failed conventional ventilation. Subject demographics, ventilator parameters, and laboratory data were extracted and analyzed. The Mann-Whitney U -test was used to assess differences in continuous variables, and the chi-square and Fisher exact tests were used for categorical variables between the groups. To assess variables that were predictive of mortality, we used both univariate and multivariate logistic regression analysis. Independent predictors of mortality were identified and used to create a multivariate risk score. Receiver operating characteristic curves were constructed to evaluate the predictive accuracy of the multivariate risk score., Results: A total of 53 premature subjects ( n = 37 male) were studied, of whom 39 (74%) survived to discharge or transfer back to referring hospital. In the univariate model, female sex, older gestational age, higher birthweight, HFJV peak inspiratory pressure at 1 h, and oxygen saturation index at 4 h were associated with mortality. In the final multivariate logistic regression model, female sex (odds ratio 4.1, 95% CI 1.2-19.8, P = .044), closed ductus arteriosus (odds ratio 7.7, 95% CI 1.3-39.5, P = .016), and oxygen saturation index > 5.5 (odds ratio 6.0, 95% CI 1.5-28.3, P = .02) were independent predictors of mortality., Conclusions: We identified that oxygen saturation index > 5.5 after 4 h of HFJV, female sex, and closed ductus arteriosus were independent predictors of mortality., Competing Interests: The authors have disclosed no conflicts of interest., (Copyright © 2020 by Daedalus Enterprises.)

Respiratory support of the critically ill neonate has steadily shifted from invasive to noninvasive forms of support. There have recently been a number of important advances in our understanding of the changes to neonatal resuscitation practices as they pertain to clinically important outcomes, mechanisms of gas exchange for high-flow nasal cannula, and best use of noninvasive ventilation and predicting response. Although the proportion of infants requiring intubation and mechanical ventilation has decreased, the most severely ill often still require intubation and ventilation. Recently, volume-targeted ventilation, high-frequency ventilation, and different methods of assessing weaning and extubation have been investigated. This review summarizes a number of important advances that have been made in the management of prematurity and neonatal respiratory distress syndrome., Competing Interests: The authors have disclosed no conflicts of interest., (Copyright © 2020 by Daedalus Enterprises.)

Background: A universal method for determining ideal body weight (IBW) for the application of appropriate tidal volumes in children on mechanical ventilation is elusive. We sought to compare 3 commonly used IBW methods for subjects between ages 2 and 20 y., Methods: Demographic data were recorded, and the IBW was calculated based on the McLaren-Read, Moore, and body mass index methods by using growth chart data from the Centers for Disease Control and Prevention. The percentage error between each IBW method and the actual body weight were calculated and reported as median (interquartile range). We decided a priori that a ≥10% difference between the actual body weight and IBW would be clinically important. The Wilcoxon signed-rank test was used to compare the actual body weight with the IBW. Bland-Altman analysis was used to assess the individual agreement of each IBW method with the actual body weight. The Kruskal-Wallis test was used to detect differences among the IBW methods., Results: A total of 58 subjects (36% female) were analyzed. The median (interquartile range) percent weight error between the actual body weight and calculated the IBW was 14.8% (1.9-22.1%, P = .038), 13.8% (4.6-23.4%, P = .008), and 12.0% (3.9-20.5%, P = .037); the mean biases were 2.7 (95% CI -13.4 to 18.9) kg, 3.9 (95% CI -15.1 to 22.9) kg, 3.2 (95% CI -16.7 to 23.1) kg; and the numbers of subjects who would have a clinically important error were 29 (55.7%), 29 (56.9%), and 30 (51.7%) for the McLaren-Read, Moore, and body mass index methods, respectively., Conclusions: The majority of the subjects demonstrated a clinically important error between the actual body weight and the IBW. The percent error increased in subjects > 25 kg actual body weight. These data underline the importance of obtaining height measurements and calculated IBW in pediatric patients who are mechanically ventilated., Competing Interests: The authors have disclosed no conflicts of interest., (Copyright © 2018 by Daedalus Enterprises.)

Background: High-frequency jet ventilation (HFJV) has been used in conjunction with conventional ventilation for infants with respiratory failure. We sought to identify parameters that were associated with successful application of HFJV in patients with hypercapnic respiratory failure., Methods: A single-center, retrospective review of infants who received HFJV was conducted. Subjects were enrolled if birthweight was ≤2,000 g and capillary P CO 2 was ≥55 mm Hg. Ventilator parameters and physiologic data were recorded at 1 h before HFJV initiation and at hours 1, 4, and 6 following conversion. Subjects were classified as responders if capillary P CO 2 was reduced by ≥10% after 1 h of HFJV. Data included peak inspiratory pressure, PEEP, capillary P CO 2 , and oxygen saturation index (equal to mean airway pressure × F IO 2 × 100/S pO 2 ). Because the data were not normally distributed, they are reported as median (interquartile range), and the Mann-Whitney test was used to assess differences in continuous data between groups. Categorical data were analyzed using a chi-square and Fisher exact test., Results: Thirty-four premature infants ( n = 24 male) were studied. Twenty-five subjects were classified as responders and demonstrated a significant reduction of capillary P CO 2 and F IO 2 and increased pH within the first hour. The non-responders demonstrated a higher conventional ventilation peak inspiratory pressure (25 cm H 2 O vs 19 cm H 2 O, P = .005) and had a greater postmenstrual age (30 weeks vs 26.5 weeks, P = .01). This group had a higher oxygen saturation index (7.25 vs 3.36, P = .03) and F IO 2 requirements (0.6 vs 0.35, P = .038) at 4 h., Conclusions: We identified that lower postmenstrual age, improvements in capillary P CO 2 and pH at 1 h, and a reduction of F IO 2 were associated with good response to HFJV. These data may help to identify patients who are likely to benefit from HFJV in the neonatal intensive care unit., Competing Interests: The authors have disclosed no conflicts of interest., (Copyright © 2017 by Daedalus Enterprises.)

The Atlantic sturgeon ( Acipenser oxyrinchus oxyrinchus ) is a long-lived, anadromous fish species ranging from Labrador, CA to Florida, USA. In the Saco River, located in the Gulf of Maine, this species was not present during a survey study ending in 1982, but was found inhabiting the estuary in 2007. Although the reason for the return of this sturgeon to this river system remains unknown, research on basic life-history information is necessary to facilitate the conservation of this federally protected species. Given the conservation status of the species, the present study used circulating sex steroid hormones to determine the sex of 288 Atlantic sturgeon captured between 2012 and 2014 in the Saco River estuary located in the Gulf of Maine. Overall, the sex was determined for 93% of Atlantic sturgeon sampled. Mean hormone values were similar to other Atlantic sturgeon reproductive studies. The findings indicate the validity of sex steroid hormones as a singular method for sex determination in wild Atlantic sturgeon. Results also indicated a likely 1:1 (male:female) sex ratio in the system, except in 2014 when a 1:3 ratio was observed. It is not believed that the Saco River estuary is used for spawning, as several impassable dams block access to spawning habitat. However, this area might provide crucial foraging for growth and development of juveniles and a habitat for adults forgoing spawning.

Elasmobranch fishes (sharks, skates, and rays) are particularly susceptible to anthropogenic threats, making a thorough understanding of their life history characteristics essential for proper management. Historically, elasmobranch reproductive data have been collected by lethal sampling, an approach that is problematic for threatened and endangered species. However, recent studies have demonstrated that non-lethal approaches can be as effective as lethal ones for assessment of the reproductive status of an animal. For example, plasma has been used to examine concentrations of steroid hormones. Additionally, skeletal muscle tissue, which can be obtained non-lethally and with minimal stress, can also be used to quantify concentrations of steroid hormones. Skeletal muscle progesterone, testosterone, and estradiol concentrations were determined to be statistically significant indicators of reproductive status in the oviparous Leucoraja erinacea, the yolk-dependent viviparous Squalus acanthias, and the yolk-sac placental viviparous Rhizoprionodon terraenovae. The results of the present study demonstrate that steroid hormones present in non-lethally harvested skeletal muscle tissue can be used as reliable indicators of reproductive status in elasmobranchs.

Background and Objectives: Lasers can be used to modify the chemical composition of dental enamel to increase the bond strength to restorative materials and to render the mineral phase more resistant to acid dissolution. Previous studies have suggested a synergistic relationship between CO(2) laser irradiation and fluoride treatment on increased resistance to acid dissolution. In this study a near-UV laser operating with lambda = 355-nm laser pulses of 3-5 nanoseconds duration was used to modify the surface morphology of dental enamel to increase the bond strength to restorative materials and increase the uptake of topical fluoride to render the surface more resistant to acid dissolution. We hypothesize that the short UV laser pulses are primarily absorbed by protein and lipid localized between the enamel prisms resulting in removal of intact mineral effectively etching the surface without thermal modification of the mineral phase. Such modification is likely to increase the permeability of the enamel surface and the subsequent absorption of fluoride. In addition, there is an increase in surface roughness without the formation of a layer of loosely adherent, thermally modified enamel that increases the bond strength to composite restorative materials., Study Design/materials and Methods: The surfaces of blocks of bovine enamel, 5 x 5 mm(2), were uniformly irradiated by 355-nm laser pulses and subsequently bonded to composite. The shear bond test was used to assess the bond strength of non-irradiated blocks (negative control), acid etched blocks (positive control), and laser irradiated blocks. The resistance to acid dissolution was evaluated using controlled surface dissolution experiments on irradiated samples, irradiated samples exposed to topical fluoride, and non-irradiated control samples with and without fluoride., Results: The laser surface treatments significantly increased the shear-bond strength of enamel to composite, to a level exceeding 20 MPa which was significantly more than the non-irradiated control samples and significantly less than the acid etch. Laser irradiation alone and topical fluoride application alone did not significantly increase the resistance to acid dissolution. The laser treatment followed by topical application of fluoride significantly increased the resistance to acid dissolution to a level of over 50% versus the control samples., Conclusions: We present a novel method for increasing bond strength to restorative materials and enhancing fluoride delivery to enamel surfaces and shed some light on the underlying mechanisms of caries inhibition via laser treatment and topical application of fluoride., (Copyright 2003 Wiley-Liss, Inc.)

Automated assays for catalase, glutathione peroxidase, glutathione reductase, and superoxide dismutase are presented. The assay for catalase is based on the peroxidatic activity of the enzyme. The glutathione peroxidase and reductase assays measure the consumption of NADPH following the reduction of t-butyl hydroperoxide and oxidized glutathione, respectively. The assay for superoxide dismutase is based on the reduction of cytochrome c. All assays utilize the Cobas FARA clinical automated analyzer and provide considerable time savings over the manual assays.

Physostigmine (PHY) is an anticholinergic drug used in the treatment of neuromuscular disorders and organophosphate poisoning. We described a sensitive, accurate, and reproducible method for PHY determination in biological materials. The method utilized a liquid/liquid, ion pair extraction, normal phase HPLC separation, and fluorometric quantitation at 240 nm excitation and 360 nm emission wavelength. We used neostigmine as a stabilizing agent to protect PHY from degradation and dimethylphysostigmine as an internal standard. The peak-height ratio vs concentration was linear over a working range from 0.50 to 25.0 ng/ml of PHY in plasma. Sensitivity of the method was 100 pg/ml of plasma which was the limit of quantitative detection under the experimental conditions used. Precision of the method was evaluated using plasma spiked with two concentrations of PHY: 1.0 and 10.0 ng/ml. Intra-day coefficient of variation (CV) ranged from 3.8 to 5.3%, and inter-day CV ranged from 1.8 to 3.6% for the two levels. The average recovery was 92%. We applied the method to examine the stability of PHY in plasma stored at -15 and -80 degrees C. The data indicated that PHY can be stored at either temperature for 9 weeks without undergoing significant alterations.

To determine if alterations in muscle morphology occur after subchronic oral administration of pyridostigmine bromide, rats were fed 90 mg/kg continuously in meal and examined at 1, 2, 4, 7, and 15 days. Within the first day, cholinesterase activity was reduced by 87% and remained inhibited by 74-91% for the entire course of the feeding. Light microscopy demonstrated that by the first day approximately 1 in 100 myofibers was shrunken and contained centralized nuclei. Electron microscopic examination showed that while presynaptic areas of neuromuscular junctions were relatively unaffected by this dose, postsynaptic areas invariably showed maximal changes. Ultrastructural alterations included disruption of myofilaments, mitochondrial changes consistent with accumulation of calcium, and nuclear alterations. These effects appeared not to be cumulative and were greatly diminished by 15 days even under constant drug administration and inhibition of cholinesterase activity. We conclude that subchronic feeding of pyridostigmine bromide induces primarily myopathic rather than neurogenic changes in the diaphragm and that some mechanism of accommodation may be activated that minimizes continued muscle injury.

Exposure to mustard-type vesicants results in alkylation of DNA and vesication. However, the biochemical mechanism for vesicant injury and whether it is localized or diffuse are not clear. We postulated that vesicant damage is mediated by free radicals, resulting in oxidative stress. These free radicals-mediated reactions may propagate systemically distal to the site of exposure. To test this hypothesis, we examined the effects of a single subcutaneous injection of the monofunctional sulfur mustard, butyl 2-chloroethyl sulfide (BCS), on the brain. We injected 3 groups (6 mice/group) of 5-month-old male, athymic, nude mice, weighing 30-35 g, subcutaneously with neat (undiluted) BCS (5 microliters/mouse). After 1, 24, and 48 h, we sacrificed the treated mice along with an untreated control group and analyzed the brains for biochemical markers of oxidative stress. Compared to untreated controls, the activity of glutathione peroxidase increased by 76%, P less than 0.005 at 24 h, and that of glutathione S-transferases by 25-37%, P less than 0.05 over the entire period. Total glutathione content in the brain was significantly lower, 17%, after 1 h and 23% after 24 h. We found also, concomitant with decreased glutathione, almost a 3-fold increase in susceptibility to lipid peroxidation. Because these changes are consistent with oxidative stress, we conclude that the effect of BCS administered subcutaneously may be translocated, reaching mouse brain, and causing oxidative stress.