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Abstract Climate-induced changes in hypoxia are among the most serious threats facing estuaries, which are among the most productive ecosystems on Earth. Future projections of estuarine hypoxia typically involve long-term multi-decadal continuous simulations or more computationally efficient time slice and delta methods that are restricted to short historical and future periods. We make a first comparison of these three methods by applying a linked terrestrial–estuarine model to the Chesapeake Bay, a large coastal-plain estuary in the eastern United States. Results show that the time slice approach accurately captures the behavior of the continuous approach, indicating a minimal impact of model memory. However, increases in mean annual hypoxic volume by the mid-twenty-first century simulated by the delta approach (+ 19%) are approximately twice as large as the time slice and continuous experiments (+ 9% and + 11%, respectively), indicating an important impact of changes in climate variability. Our findings suggest that system memory and projected changes in climate variability, as well as simulation length and natural variability of system hypoxia, should be considered when deciding to apply the more computationally efficient delta and time slice methods.

Pediatric traumatic brain injury (pTBI) is a major risk factor associated with adulthood incarceration. Most research into the link between pTBI and adulthood incarceration has focused on incarcerated males, who comprise the vast majority of incarcerated adults, particularly in industrialized nations. In this review, we sought to identify sex-related differences in the incidence and pathophysiology of pTBI and subsequent risk of adulthood incarceration. A scoping review was undertaken using PubMed, Scopus, Ovid, and the Cochrane Library. Articles analyzing sex-related differences in pTBI and adult incarceration rates, studies conducted on an incarcerated population, and cohort studies, cross-sectional studies, clinical trials, systematic reviews, or meta-analyses were included in this review. Of the 85 unique results, 25 articles met our inclusion criteria. Male children are 1.5 times more likely to suffer a TBI than females; however, the prevalence of incarcerated adults with a history of pTBI is ?35?45% for both sexes. Neurophysiologically, female sex hormones are implicated in neuroprotective roles, mitigating central nervous system (CNS) damage post-TBI, although this role may be more complex, given that injury severity and sequelae have been correlated with male sex whereas increased mortality has been correlated with female sex. Further investigation into the relationship between estrogen and subsequent clinical measurements of CNS function is needed to develop interventions that may alleviate the pathophysiological consequences of pTBI.

Though surface water quality has been monitored in southern Peru over the past and current century, it has been implemented by multiple organizations. The data lacks a centralized repository and access requires logistical and temporal hurdles associated with official requests. A substantial portion of the data has not been quality assured and is in difficult-to-access formats such as scanned PDF documents. These obstacles collectively make it challenging to maximize the impact of these monitoring efforts such as efficiently evaluating long-term water quality trends. To address this opportunity, we gathered available surface water quality information from five watersheds in the Arequipa Region of southern Peru: Camaná, Chili, Ocoña, Tambo, and Yauca. The effort required entry of more than 130,000 records of water quality properties across 274 monitoring stations with data including the concentration of select nutrients, metals, organic compounds, and biological taxa. The water quality records in the Chili watershed go back as far as 1905, while data for the other watersheds was largely confined to the years 2012–2021. This document describes how the surface water quality information was assimilated with quality control and provides a centralized Excel database so that the data can be efficiently used for research and decision making purposes.

Infectious hematopoietic necrosis (IHN) is a disease of salmonid fish that is caused by the IHN virus (IHNV), which can cause substantial mortality and economic losses in rainbow trout aquaculture and fisheries enhancement hatchery programs. In a previous study on a commercial rainbow trout breeding line that has undergone selection, we found that genetic resistance to IHNV is controlled by the oligogenic inheritance of several moderate and many small effect quantitative trait loci (QTL). Here we used genome wide association analyses in two different commercial aquaculture lines that were naïve to previous exposure to IHNV to determine whether QTL were shared across lines, and to investigate whether there were major effect loci that were still segregating in the naïve lines. A total of 1,859 and 1,768 offspring from two commercial aquaculture strains were phenotyped for resistance to IHNV and genotyped with the rainbow trout Axiom 57K SNP array. Moderate heritability values (0.15–0.25) were estimated. Two statistical methods were used for genome wide association analyses in the two populations. No major QTL were detected despite the naïve status of the two lines. Further, our analyses confirmed an oligogenic architecture for genetic resistance to IHNV in rainbow trout. Overall, 17 QTL with notable effect (≥1.9% of the additive genetic variance) were detected in at least one of the two rainbow trout lines with at least one of the two statistical methods. Five of those QTL were mapped to overlapping or adjacent chromosomal regions in both lines, suggesting that some loci may be shared across commercial lines. Although some of the loci detected in this GWAS merit further investigation to better understand the biological basis of IHNV disease resistance across populations, the overall genetic architecture of IHNV resistance in the two rainbow trout lines suggests that genomic selection may be a more effective strategy for genetic improvement in this trait.

Abstract As an abundant element in the Earth’s crust, sodium plays an unusual role in food webs. Its availability in terrestrial environments is highly variable, but it is nonessential for most plants, yet essential for animals and most decomposers. Accordingly, sodium requirements are important drivers of various animal behavioural patterns and performance levels. To specifically test whether sodium limitation increases cannibalism in a gregarious lepidopteran herbivore, we hydroponically manipulated Helianthus annuus host plants' tissue-sodium concentrations. Gregarious larvae of the bordered patch butterfly, Chlosyne lacinia, cannibalized siblings when plant-tissue sodium concentrations were low in two separate experiments. Although cannibalism was almost non-existent when sodium concentrations were high, individual mortality rates were also high. Sodium concentration in host plants can have pronounced effects on herbivore behaviour, individual-level performance, and population demographics, all of which are important for understanding the ecology and evolution of plant-animal interactions across a heterogeneous phytochemical landscape.

Polar ring galaxies (PRGs) are unusual relative to common galaxies in that they consist of a central host galaxy—usually a gas-poor, early-type S0 or elliptical galaxy—surrounded by a ring of gas, dust and stars that orbit perpendicular to the major axis of the host. Despite the general quiescence of early-type galaxies (ETGs) and the rings’ lack of spiral density waves, PRGs are the sites of significant star formation relative to typical ETGs. To study these structures and improve PRG statistics, we obtained and analyzed infrared (IR) images from the Infrared Array Camera (IRAC) aboard the Spitzer Space Telescope, and combined these IR data with archival optical data from both the Sloan Digital Sky Survey and the Hubble Space Telescope, and with optical imaging data we obtained with the Gemini South Observatory. We performed structural decomposition and photometry for five PRGs, and fit the spectral energy distributions (SEDs) of each PRG component to estimate the stellar masses, ages, and other physical properties of the PRG components. We show that PRC B-12 and PRC B-22, both lacking previous analysis, obey trends commonly observed among PRGs. We find that the stellar masses of polar rings can be a significant fraction of the host galaxy’s stellar masses (∼10–30%). We note, however, that our estimates of stellar mass and other physical properties are the results of SED fitting and not direct measurements. Our findings corroborate both previous theoretical expectations and measurements of existing samples of PRGs and indicate the utility of SED fitting in the context of these unusual galaxies, which historically have lacked multi-wavelength photometry of their stellar components. Finally, we outline future improvements needed for more definitive studies of PRGs and their formation scenarios.

IntroductionMany marine organisms have a biphasic life cycle that transitions between a swimming larva with a more sedentary adult form. At the end of the first phase, larvae must identify suitable sites to settle and undergo a dramatic morphological change. Environmental factors, including photic and chemical cues, appear to influence settlement, but the sensory receptors involved are largely unknown. We targeted the protein receptor, opsin, which belongs to large superfamily of transmembrane receptors that detects environmental stimuli, hormones, and neurotransmitters. While opsins are well-known for light-sensing, including vision, a growing number of studies have demonstrated light-independent functions. We therefore examined opsin expression in the Pteriomorphia, a large, diverse clade of marine bivalves, that includes commercially important species, such as oysters, mussels, and scallops.MethodsGenomic annotations combined with phylogenetic analysis show great variation of opsin abundance among pteriomorphian bivalves, including surprisingly high genomic abundance in many species that are eyeless as adults, such as mussels. Therefore, we investigated the diversity of opsin expression from the perspective of larval development. We collected opsin gene expression in four families of Pteriomorphia, across three distinct larval stages, i.e., trochophore, veliger, and pediveliger, and compared those to adult tissues.ResultsWe found larvae express all opsin types in these bivalves, but opsin expression patterns are largely species-specific across development. Few opsins are expressed in the adult mantle, but many are highly expressed in adult eyes. Intriguingly, opsin genes such as retinochrome, xenopsins, and Go-opsins have higher levels of expression in the later larval stages when substrates for settlement are being tested, such as the pediveliger.ConclusionInvestigating opsin gene expression during larval development provides crucial insights into their intricate interactions with the surroundings, which may shed light on how opsin receptors of these organisms respond to various environmental cues that play a pivotal role in their settlement process.

The purpose of this retrospective case series was to review the demographics of alpine skiers who sustain tibial plateau fractures, evaluate the inter-observer reliability of the Schatzker classification for fractures sustained while alpine skiing, and to evaluate patient-reported outcomes. We hypothesized that most tibial plateau fractures would be low-energy fracture patterns (Schatzker I–III) and occur in women and less-experienced skiers. Charts and radiographs of patients treated surgically for tibial plateau fractures caused by alpine skiing were evaluated. Patients treated less than two years prior to review were excluded. Patients who qualified were contacted to complete a questionnaire regarding their skiing experience, their pain levels experienced following their accident, the weather conditions during the accident, and their return to sport. Forty-seven patients met the inclusion criteria. The mean age was 49, and 60% of the participants were male. There were 28 low-energy fracture patterns (Schatzker I–III) and 19 high-energy patterns (Schatzker IV–VI) with a 95.7% inter-observer reliability. Contrary to the hypothesis, tibial plateau fractures sustained while alpine skiing occurred in older, experienced riders. Approximately 40% were high-energy fractures. Although over 75% of patients reported having no pain or occasional pain at their final follow-ups, less than half of the patients returned to alpine skiing.

Abstract Introduction Racial and language disparities in the United States healthcare system have long undermined the quality of care provided to minority patients. With the projected growth of the Hispanic population, there is an urgent need for medical schools to integrate high-quality medical Spanish and cultural competency content. We propose a comprehensive medical Spanish curriculum aligned with the preclinical curriculum as a solution to these issues. The primary goal of this study is to demonstrate the effectiveness of a clinically focused, culturally competent medical Spanish program and advocate for its widespread adoption in medical institutions nationwide. Methods The study utilized the Kirkpatrick Model to evaluate the success of the medical Spanish curriculum. A total of 111 medical students voluntarily enrolled in the medical Spanish course. Out of these students, 47 completed the final evaluation, which included a Spanish Objective Structured Clinical Examination and a 40-question Multiple-Choice Exam assessing the integration of Spanish language skills and cultural competency. Both assessment methods took place in clinical skills facilities. Descriptive statistics summarized exam results, and two-tailed t-tests compared mean exam scores between students of different proficiency levels. Results and discussion Students achieved a mean score of over 80% on all components of the Spanish Objective Structured Clinical Examination and the Multiple-Choice Exam. Survey data suggest that students felt able to communicate in Spanish with patients after completing the course series. The study also provides a model for a medical Spanish curriculum that applies expert-recommended best practices to meet the needs of Hispanic patient populations. Limitations and conclusions Students who sat for the OSCE and MCE were self-selected. Baseline data on student perceptions and Spanish competency are not sufficient for making comparisons.

Several models for estimating sporadic-E intensity from Global Navigation Satellite System (GNSS) radio occultation (RO) observation have previously been developed using a single perturbation or intensity parameter, such as phase-based total electron content (TEC) or the amplitude-based S4 index. Here, we outline two new models that use a combination of phase and amplitude parameters for the L1 and L2 signals. These models show a significant improvement over the baseline models used for comparison. Furthermore, the GNSS-RO parameters are compared with several different ionosonde intensity parameters including the direct foEs and fbEs measurements along with the metallic-ion based foμEs and fbμEs parameters which account for the background E-region density. Interestingly, the phase-based σϕ scintillation index shows the strongest correlation to foEs and fbEs while amplitude-based S4 shows the strongest correlation to foμEs and fbμEs. While the metallic-ion based foμEs and fbμEs parameters are physically ideal for GNSS-RO observations, we show difficulties in practical implementation due to the reliance on a background E-region density estimate using a model such as the International Reference Ionosphere (IRI). Ultimately, we provide two improved sporadic-E intensity models that can be used for future GNSS-RO based studies along with a recommendation to compare against the ionosonde-based foEs parameter.

Curative bone marrow transplantation (BMT) therapies for sickle cell disease (SCD) can cause infertility. The Fertility Preservation Program (FPP) in Pittsburgh cryopreserves testicular tissues for SCD patients prior to BMT in anticipation that those tissues can be thawed in the future and matured to produce sperm. Here, we generated and validated two isogenic patient-derived induced pluripotent stem cell (iPSC) lines from testicular biopsy fibroblasts of a 12-year-old SCD patient.

Abstract The effects of warming on ecological communities emerge from a range of potentially asymmetric impacts on individual physiology and development. Understanding these responses, however, is limited by our ability to connect mechanisms or emergent patterns across the many processes that drive variation in demography. Further complicating this understanding is the gain or loss of predators to many communities, which may interact with changes in temperature to drive community change. Here we conducted a factorial warming and predation experiment to test generalized predictions about responses to warming. We used microcosms with a range of protists, rotifers, and a gastrotrich, with and without the predator Actinosphaerium, to assess changes in diversity, body size, function, and composition in response to warming. We find that community respiration and predator:prey biovolume ratios peak at intermediate temperatures, while species richness declined with temperature. We also found that overall biomass increased with species richness, driven by the effect of temperature on richness. There was little evidence of an interaction between predation and temperature change, likely because the predator was mostly limited to the intermediate temperatures. Overall, our results suggest that general predictions about community change are still challenging to make but may benefit by considering multiple dimensions of community patterns in an integrated way.

Abstract Some mistletoe species (Loranthaceae) resemble their host plants to a striking degree. Various mechanisms have been proposed for the developmental origins of novel traits that cause mistletoes to appear similar to their hosts, as well as for the adaptive phenotypic evolution of such traits. Calder (1983) proposed a logically flawed group selectionist seed‐dispersal hypothesis for mistletoes to resemble their hosts. Calder's (1983) hypothesis does not provide a viable potential explanation for mistletoe resemblance to hosts.

Female cancer patients who have undergone chemotherapy have an elevated risk of developing ovarian dysfunction and failure. Experimental approaches to treat iatrogenic infertility are evolving rapidly; however, challenges and risks remain that hinder clinical translation. Biomaterials have improved in vitro follicle maturation and in vivo transplantation in mice, but there has only been marginal success for early-stage human follicles. Here, we developed methods to obtain an ovarian-specific extracellular matrix hydrogel to facilitate follicle delivery and establish an in situ ovary (ISO), which offers a permissive environment to enhance follicle survival. We demonstrate sustainable follicle engraftment, natural pregnancy, and the birth of healthy pups after intraovarian microinjection of isolated exogenous follicles into chemotherapy-treated (CTx) mice. Our results confirm that hydrogel-based follicle microinjection could offer a minimally invasive delivery platform to enhance follicle integration for patients post-chemotherapy.

Abstract Background Cellular entry of SARS-CoV-2 has been shown to rely on angiotensin-converting enzyme 2 (ACE2) receptors, whose expression in the testis is among the highest in the body. Additionally, the risk of mortality seems higher among male COVID-19 patients, and though much has been published since the first cases of COVID-19, there remain unanswered questions regarding SARS-CoV-2 impact on testes and potential consequences for reproductive health. We investigated testicular alterations in non-vaccinated deceased COVID-19-patients, the precise location of the virus, its replicative activity, and the immune, vascular, and molecular fluctuations involved in the pathogenesis. Results We found that SARS-CoV-2 testicular tropism is higher than previously thought and that reliable viral detection in the testis requires sensitive nanosensors or RT-qPCR using a specific methodology. Through an in vitro experiment exposing VERO cells to testicular macerates, we observed viral content in all samples, and the subgenomic RNA’s presence reinforced the replicative activity of SARS-CoV-2 in testes of the severe COVID-19 patients. The cellular structures and viral particles, observed by transmission electron microscopy, indicated that macrophages and spermatogonial cells are the main SARS-CoV-2 lodging sites, where new virions form inside the endoplasmic reticulum Golgi intermediate complex. Moreover, we showed infiltrative infected monocytes migrating into the testicular parenchyma. SARS-CoV-2 maintains its replicative and infective abilities long after the patient’s infection. Further, we demonstrated high levels of angiotensin II and activated immune cells in the testes of deceased patients. The infected testes show thickening of the tunica propria, germ cell apoptosis, Sertoli cell barrier loss, evident hemorrhage, angiogenesis, Leydig cell inhibition, inflammation, and fibrosis. Conclusions Our findings indicate that high angiotensin II levels and activation of mast cells and macrophages may be critical for testicular pathogenesis. Importantly, our findings suggest that patients who become critically ill may exhibit severe alterations and harbor the active virus in the testes.

Despite over two decades of progress against HIV/AIDS in adjacent sub-Saharan Africa, HIV rates and deaths due to AIDS are exponentially rising in Madagascar. Furthermore, a growing body of evidence suggests that, due to a scarcity of general-population screening data, even the startling increase demonstrated by official models vastly underestimates the true population prevalence of HIV. We aimed to implement a real-world HIV screening and treatment protocol to serve a general population stemming from across northern Madagascar. In collaboration with the Malagasy Ministry of Health, we provided point-of-care HIV screening and confirmatory testing for over 1000 participants from 73 towns, villages, and cities. We recorded an overall HIV prevalence of 2.94%. Notably, we observed a 13.1% HIV prevalence rate among urban populations and showed that proximity to a major route of travel was significantly associated with HIV risk. We also observed a link between HIV risk and various occupations, including those associated with increased mobility (such as mining). Importantly, all HIV-positive individuals were initiated on antiretroviral therapy in concordance with local health authorities. To our knowledge, this study marks the largest primary test data-based HIV study to date among Madagascar’s general population, showing a greatly higher HIV prevalence (2.9%) than previously reported modeling-based figures (0.4%). Our rates aligned with the pattern of higher prevalence demonstrated in smaller general-population screening studies occurring more commonly prior to political strife in the mid-2000s. These findings demonstrate evidence of a growing HIV epidemic in northern Madagascar and underscore the need for future investment into more comprehensive HIV screening and control initiatives in Madagascar.

BackgroundCryopreservation of immature testicular tissue (ITT) is currently the only option to preserve fertility of prepubertal patients. Autologous transplantation of ITT may not be safe or appropriate for all patients. Therefore, methods to mature ITT ex vivo are needed.ObjectivesAim to investigate the feasibility of inducing in vitro spermatogenesis from ITT cryopreserved for pediatric patients prior to initiation of gonadotoxic therapy.Materials and methodsCryopreserved-thawed ITT from prepubertal and peripubertal patients were cultured for 7, 16, and 32 days in medium with no hormones or supplemented with 5 IU/L FSH, 1 IU/L hCG, or 5IU/L FSH+1 IU/L hCG. Samples were evaluated histologically to assess tissue integrity, and immunofluorescence staining was performed to identify VASA (DDX4)+ germ cells, UCHL1+ spermatogonia, SYCP3+ spermatocytes, CREM+ spermatids, SOX9+ Sertoli cells. Proliferation (KI67) and apoptosis (CASPASE3) of germ cells and Sertoli cells were also analyzed. Sertoli and Leydig cell maturation was evaluated by AR and INSL3 expression as well as expression of the blood testis barrier protein, CLAUDIN11, and testosterone secretion in the culture medium.ResultsIntegrity of seminiferous tubules, VASA+ germ cells and SOX9+ Sertoli cells were maintained up to 32 days. The number of VASA+ germ cells was consistently higher in the peripubertal groups. UCHL1+ undifferentiated spermatogonia and SOX9+ Sertoli cell proliferation was confirmed in most samples. SYCP3+ primary spermatocytes began to appear by day 16 in both age groups. Sertoli cell maturation was demonstrated by AR expression but the expression of CLAUDIN11 was disorganized. Presence of mature and functional Leydig cells was verified by INSL3 expression and secretion of testosterone. Gonadotropin treatments did not consistently impact the number or proliferation of germ cells or somatic cells, but FSH was necessary to increase testosterone secretion over time in prepubertal samples.ConclusionITT were maintained in organotypic culture for up to 32 days and spermatogonia differentiated to produce primary spermatocytes in both pre- and peripubertal age groups. However, complete spermatogenesis was not observed in either group.

Summary: Nutrient starvation drives yeast meiosis, whereas retinoic acid (RA) is required for mammalian meiosis through its germline target Stra8. Here, by using single-cell transcriptomic analysis of wild-type and Stra8-deficient juvenile mouse germ cells, our data show that the expression of nutrient transporter genes, including Slc7a5, Slc38a2, and Slc2a1, is downregulated in germ cells during meiotic initiation, and this process requires Stra8, which binds to these genes and induces their H3K27 deacetylation. Consequently, Stra8-deficient germ cells sustain glutamine and glucose uptake in response to RA and exhibit hyperactive mTORC1/protein kinase A (PKA) activities. Importantly, expression of Slc38a2, a glutamine importer, is negatively correlated with meiotic genes in the GTEx dataset, and Slc38a2 knockdown downregulates mTORC1/PKA activities and induces meiotic gene expression. Thus, our study indicates that RA via Stra8, a chordate morphogen pathway, induces meiosis partially by generating a conserved nutrient restriction signal in mammalian germ cells by downregulating their nutrient transporter expression.

Summary: Spermatogonial stem cells (SSCs) in the testis support the lifelong production of sperm. SSCs reside within specialized microenvironments called “niches,” which are essential for SSC self-renewal and differentiation. However, our understanding of the molecular and cellular interactions between SSCs and niches remains incomplete. Here, we combine spatial transcriptomics, computational analyses, and functional assays to systematically dissect the molecular, cellular, and spatial composition of SSC niches. This allows us to spatially map the ligand-receptor (LR) interaction landscape in both mouse and human testes. Our data demonstrate that pleiotrophin regulates mouse SSC functions through syndecan receptors. We also identify ephrin-A1 as a potential niche factor that influences human SSC functions. Furthermore, we show that the spatial re-distribution of inflammation-related LR interactions underlies diabetes-induced testicular injury. Together, our study demonstrates a systems approach to dissect the complex organization of the stem cell microenvironment in health and disease.

Abstract Understanding the phytochemical landscapes of essential and nonessential chemical elements to plants provides an opportunity to better link biogeochemical cycles to trophic ecology. We investigated the formation and regulation of the cationic phytochemical landscapes of four key elements for biota: Ca, Mg, K, and Na. We collected aboveground tissues of plants in Atriplex, Helianthus, and Opuntia and adjacent soils from 51, 131, and 83 sites, respectively, across the southern United States. We determined the spatial variability of these cations in plants and soils. Also, we quantified the homeostasis coefficient for each cation and genus combination, by using mixed‐effect models, with spatially correlated random effects. Additionally, using random forest models, we modeled the influence of bioclimatic, soil, and spatial variables on plant cationic concentrations. Sodium variability and spatial autocorrelation were considerably greater than for Ca, Mg, or K. Calcium, Mg, and K exhibited strongly homeostatic patterns, in striking contrast to non‐homeostatic Na. Even so, climatic and soil variables explained a large proportion of plants' cationic concentrations. Essential elements (Ca, Mg, and K) appeared to be homeostatically regulated, which contrasted sharply with Na, a nonessential element for most plants. In addition, we provide evidence for the No‐Escape‐from‐Sodium hypothesis in real‐world ecosystems, indicating that plant Na concentrations tend to increase as substrate Na levels increase.

Atmospheric water harvesting by MOF-based devices is critically dependent on weather conditions. Here, the authors report a MOF-based adaptive water harvesting system capable of adapting its cycles to temperature and relative humidity fluctuations throughout the day, continuously optimizing water production.

The healthy skeletal muscle extracellular matrix (ECM) has several functions including providing structural integrity to myofibers, enabling lateral force transmission, and contributing to overall passive mechanical properties. In diseases such as Duchenne Muscular dystrophy, there is accumulation of ECM materials, primarily collagen, which results in fibrosis. Previous studies have shown that fibrotic muscle is often stiffer than healthy muscle, in part due to the increased number and altered architecture of collagen fibers within the ECM. This would imply that the fibrotic matrix is stiffer than the healthy matrix. However, while previous studies have attempted to quantify the extracellular contribution to passive stiffness in muscle, the outcomes are dependent on the type of method used. Thus, the goals of this study were to compare the stiffness of healthy and fibrotic muscle ECM and to demonstrate the efficacy of two methods for quantifying extracellular-based stiffness in muscle, namely decellularization and collagenase digestion. These methods have been demonstrated to remove the muscle fibers or ablate collagen fiber integrity, respectively, while maintaining the contents of the extracellular matrix. Using these methods in conjunction with mechanical testing on wildtype and D2.mdx mice, we found that a majority of passive stiffness in the diaphragm is dependent on the ECM, and the D2.mdx diaphragm ECM is resistant to digestion by bacterial collagenase. We propose that this resistance is due to the increased collagen cross-links and collagen packing density in the ECM of the D2.mdx diaphragm. Taken altogether, while we did not find increased stiffness of the fibrotic ECM, we did observe that the D2.mdx diaphragm conveyed resistance against collagenase digestion. These findings demonstrate how different methods for measuring ECM-based stiffness each have their own limitations and can produce different results.

Introduction: Human spermatogenesis is a highly intricate process that requires the input of thousands of testis-specific genes. Defects in any of them at any stage of the process can have detrimental effects on sperm production and/or viability. In particular, the function of many meiotic proteins encoded by germ cell specific genes is critical for maturation of haploid spermatids and viable spermatozoa, necessary for fertilization, and is also extremely sensitive to even the slightest change in coding DNA.Methods: Here, using whole exome and genome approaches, we identified and reported novel, clinically significant variants in testis-expressed gene 15 (TEX15), in unrelated men with spermatogenic failure (SPGF).Results: TEX15 mediates double strand break repair during meiosis. Recessive loss-of-function (LOF) TEX15 mutations are associated with SPGF in humans and knockout male mice are infertile. We expand earlier reports documenting heterogeneous allelic pathogenic TEX15 variants that cause a range of SPGF phenotypes from oligozoospermia (low sperm) to nonobstructive azoospermia (no sperm) with meiotic arrest and report the prevalence of 0.6% of TEX15 variants in our patient cohort. Among identified possible LOF variants, one homozygous missense substitution c.6835G>A (p.Ala2279Thr) co-segregated with cryptozoospermia in a family with SPGF. Additionally, we observed numerous cases of inferred in trans compound heterozygous variants in TEX15 among unrelated individuals with varying degrees of SPGF. Variants included splice site, insertions/deletions (indels), and missense substitutions, many of which resulted in LOF effects (i.e., frameshift, premature stop, alternative splicing, or potentially altered posttranslational modification sites).Conclusion: In conclusion, we performed an extensive genomic study of familial and sporadic SPGF and identified potentially damaging TEX15 variants in 7 of 1097 individuals of our combined cohorts. We hypothesize that SPGF phenotype severity is dictated by individual TEX15 variant’s impact on structure and function. Resultant LOFs likely have deleterious effects on crossover/recombination in meiosis. Our findings support the notion of increased gene variant frequency in SPGF and its genetic and allelic heterogeneity as it relates to complex disease such as male infertility.

The manchette is a transient and unique structure present in elongating spermatids and required for proper differentiation of the germ cells during spermatogenesis. Previous work indicated that the MEIG1/PACRG complex locates in the manchette and is involved in the transport of cargos, such as SPAG16L, to build the sperm flagellum. Here, using co-immunoprecipitation and pull-down approaches in various cell systems, we established that DNALI1, an axonemal component originally cloned from Chlamydomonas reinhardtii, recruits and stabilizes PACRG and we confirm in vivo, the co-localization of DNALI1 and PACRG in the manchette by immunofluorescence of elongating murine spermatids. We next generated mice with a specific deficiency of DNALI1 in male germ cells, and observed a dramatic reduction of the sperm cells, which results in male infertility. In addition, we observed that the majority of the sperm cells exhibited abnormal morphology including misshapen heads, bent tails, enlarged midpiece, discontinuous accessory structure, emphasizing the importance of DNALI1 in sperm differentiation. Examination of testis histology confirmed impaired spermiogenesis in the mutant mice. Importantly, while testicular levels of MEIG1, PACRG, and SPAG16L proteins were unchanged in the Dnali1 mutant mice, their localization within the manchette was greatly affected, indicating that DNALI1 is required for the formation of the MEIG1/PACRG complex within the manchette. Interestingly, in contrast to MEIG1 and PACRG-deficient mice, the DNALI1-deficient mice also showed impaired sperm spermiation/individualization, suggesting additional functions beyond its involvement in the manchette structure. Overall, our work identifies DNALI1 as a protein required for sperm development.

Rainbow trout lines have been selected to produce gametes at different times of the year to insure a year-round supply of seedstock. Protocols for cryopreservation of milt have also been established and cryopreservation services are currently commercially available, further extending the availability of gametes. The present study was conducted to determine the impact of crossing rainbow trout with different spawning seasons on the spawning date and gamete characteristics of the offspring. Egg lots from females from an August-spawning population were divided in half and fertilized with milt from males from the same population (‘purebred’) or cryopreserved milt from two males from a February-spawning population (‘hybrid’). The purebred and hybrid groups both exhibited spawning windows that lasted around three months with each family taking about nine weeks for the middle 10–90% of the individual females to ovulate. However, the hybrid progeny spawned on average about 10 weeks after the purebred families, peaking in November and thus exhibiting a classic phenotypic distribution pattern of heterozygous F1 hybrids. The hybrid progeny did not spawn the next spring which was the alternative intermediate to February and August. The F1 hybrids were crossed to produce an F2 generation for future genetic analysis to map quantitative trait loci (QTL) for spawning date. Consistent with the February line historically producing larger eggs than the August line, the hybrid offspring produced slightly larger eggs than the purebred August line families, as well as yielding a greater volume of eggs per unit body weight. These results are valuable for strategies to maximize hatchery production of year-round seedstock and for QTL mapping of spawning date in rainbow trout.

Abstract Botulinum neurotoxins (BoNTs) are highly potent, select agent toxins that inhibit neurotransmitter release at motor nerve terminals, causing muscle paralysis and death by asphyxiation. Other than post-exposure prophylaxis with antitoxin, the only treatment option for symptomatic botulism is intubation and supportive care until recovery, which can require weeks or longer. In previous studies, we reported the FDA-approved drug 3,4-diaminopyridine (3,4-DAP) reverses early botulism symptoms and prolongs survival in lethally intoxicated mice. However, the symptomatic benefits of 3,4-DAP are limited by its rapid clearance. Here we investigated whether 3,4-DAP could sustain symptomatic benefits throughout the full course of respiratory paralysis in lethally intoxicated rats. First, we confirmed serial injections of 3,4-DAP stabilized toxic signs and prolonged survival in rats challenged with 2.5 LD50 BoNT/A. Rebound of toxic signs and death occurred within hours after the final 3,4-DAP treatment, consistent with the short half-life of 3,4-DAP in rats. Based on these data, we next investigated whether the therapeutic benefits of 3,4-DAP could be sustained throughout the course of botulism by continuous infusion. To ensure administration of 3,4-DAP at clinically relevant doses, three infusion dose rates (0.5, 1.0 and 1.5 mg/kg∙h) were identified that produced steady-state serum levels of 3,4-DAP consistent with clinical dosing. We then compared dose-dependent effects of 3,4-DAP on toxic signs and survival in rats intoxicated with 2.5 LD50 BoNT/A. In contrast to saline vehicle, which resulted in 100% mortality, infusion of 3,4-DAP at ≥ 1.0 mg/kg∙h from 1 to 14 d after intoxication produced 94.4% survival and full resolution of toxic signs, without rebound of toxic signs after infusion was stopped. In contrast, withdrawal of 3,4-DAP infusion at 5 d resulted in re-emergence of toxic sign and death within 12 h, confirming antidotal outcomes require sustained 3,4-DAP treatment for longer than 5 d after intoxication. We exploited this novel survival model of lethal botulism to explore neurophysiological parameters of diaphragm paralysis and recovery. While neurotransmission was nearly eliminated at 5 d, neurotransmission was significantly improved at 21 d in 3,4-DAP-infused survivors, although still depressed compared to naïve rats. 3,4-DAP is the first small molecule to reverse systemic paralysis and promote survival in animal models of botulism, thereby meeting a critical treatment need that is not addressed by post-exposure prophylaxis with conventional antitoxin. These data contribute to a growing body of evidence supporting the use of 3,4-DAP to treat clinical botulism.

Abstract The role of ER Ca2+ release via ryanodine receptors (RyR) in pancreatic β-cell function is not well defined. Deletion of RyR2 from the rat insulinoma INS-1 (RyR2KO) enhanced IP3 receptor activity stimulated by 7.5 mM glucose, coincident with reduced levels of the protein IP3 Receptor Binding protein released with Inositol 1,4,5 Trisphosphate (IRBIT). Insulin content, basal (2.5 mM glucose) and 7.5 mM glucose-stimulated insulin secretion were reduced in RyR2KO and IRBITKO cells compared to controls. INS2 mRNA levels were reduced in both RyR2KO and IRBITKO cells, but INS1 mRNA levels were specifically decreased in RyR2KO cells. Nuclear localization of S-adenosylhomocysteinase (AHCY) was increased in RyR2KO and IRBITKO cells. DNA methylation of the INS1 and INS2 gene promotor regions was very low, and not different among RyR2KO, IRBITKO, and controls, but exon 2 of the INS1 and INS2 genes was more extensively methylated in RyR2KO and IRBITKO cells. Exploratory proteomic analysis revealed that deletion of RyR2 or IRBIT resulted in differential regulation of 314 and 137 proteins, respectively, with 41 in common. These results suggest that RyR2 regulates IRBIT levels and activity in INS-1 cells, and together maintain insulin content and secretion, and regulate the proteome, perhaps via DNA methylation.

Introduction Treatment of non‐acute subdural hematoma (NASDH) remains challenging due to recurrence ranging from 2–37%. Middle meningeal artery embolization (MMAe) has emerged as a minimally invasive procedure uniquely poised to improve outcomes in NASDH. The goal of this study is to evaluate the clinical and radiographic characteristics of patients undergoing MMAe for NASDH, and how outcomes for MMAe differ when utilized upfront, prophylactically, or as a salvage modality after surgical evacuation. Furthermore, we investigated how independent patient risk factors, such as oncologic history and anticoagulant/antiplatelet (ACAP) medication use, may impact MMAe and NASDH outcomes. Methods This prospective study followed patients undergoing MMAe for NASDH from 2016–2021. All patients were diagnosed with a NASDH on non‐contrast CT imaging and underwent MMAe by the lead neurosurgeon. The primary outcome was NASDH recurrence post‐MMAe requiring surgical evacuation, repeat MMAe, or both within the one‐year follow up period. Additional radiographic outcomes included reduction of hematoma width and midline shift at longest follow‐up. Subgroup analyses stratified outcomes by MMAe indication: upfront (previously untreated and nonoperative NASDH), prophylactic (MMAe 1–5 days after surgical evacuation), or salvage (individuals with radiographic and clinical NASDH recurrence). Outcomes among oncologic, coagulopathic, and ACAP patients were also assessed. The modified Rankin scale (mRS) quantified post‐MMAe clinical outcomes. Results A total of 236 NASDH patients underwent 294 MMAe. Of these, 115 (48.7%), 92 (39.0%), and 29 (12.3%) patients received upfront, prophylactic, and salvage MMAe, respectively. Upfront MMAe was performed more frequently than prophylactic and salvage for patients with co‐existing malignancy (29.6%, 12.0%, 20.7%; P < 0.01) and coagulopathy (17.4%, 4.3%, 10.3%; P = 0.01). Prophylactic MMAe was more commonly performed for subacute SDH compared with upfront or salvage (15.2%, 2.6%, 0%; P < 0.001); no differences in utilization were seen for chronic or acute on chronic SDH. Sixteen (5.4%) patients presented with hematoma recurrence after MMAe requiring repeat intervention, of which 5 (1.7%) received a craniotomy, 7 (2.4%) received repeat MMAe, and 4 (1.3%) underwent both MMAe and surgical evacuation. Risk of overall NASDH recurrence requiring re‐intervention did not differ among prophylactic, upfront, and salvage groups (6.5%, 7.0%, 6.9%; P > 0.99). Clinical outcomes did not differ between MMAe indications at longest follow‐up. Within the prophylactic group, radiographic and clinical outcomes were similar between MMAe paired with twist‐drill craniostomy or with craniotomy. Subgroup analysis revealed MMAe also effectively decreased NASDH recurrence in ACAP, oncologic, and coagulopathic patients. However, patients with both an oncologic history and concurrent ACAP use less frequently achieved improved clinical status at follow‐up compared with those harboring ACAP/coagulopathic history alone, oncologic history alone, or neither (31.4%, 45.9%, 50.0%, 57.3%; P = 0.049). Conclusions MMAe is emerging as a viable treatment for curtailing recurrence and improving NASDH outcomes, with similar clinical outcomes when performed prophylactically, upfront, or as salvage treatment. NASDH patients presenting with an oncologic history and concurrent ACAP use may be less likely to achieve improved clinical status after MMAe.

The ER Ca2+ channel ryanodine receptor 2 (RyR2) is required for maintenance of insulin content and glucose-stimulated insulin secretion, in part, via regulation of the protein IRBIT in the insulinoma cell line INS-1. Here, we examined store-operated and depolarization-dependent Ca2+entry using INS-1 cells in which either RyR2 or IRBIT were deleted. Store-operated Ca2+ entry (SOCE) stimulated with thapsigargin was reduced in RyR2KO cells compared to controls, but was unchanged in IRBITKO cells. STIM1 protein levels were not different between the three cell lines. Basal and stimulated (500 μM carbachol) phospholipase C (PLC) activity was also reduced specifically in RyR2KO cells. Insulin secretion stimulated by tolbutamide was reduced in RyR2KO and IRBITKO cells compared to controls, but was potentiated by an EPAC-selective cAMP analog in all three cell lines. Cellular PIP2 levels were increased and cortical f-actin levels were reduced in RyR2KO cells compared to controls. Whole-cell Cav channel current density was increased in RyR2KO cells compared to controls, and barium current was reduced by acute activation of the lipid phosphatase pseudojanin preferentially in RyR2KO cells over control INS-1 cells. Action potentials stimulated by 18 mM glucose were more frequent in RyR2KO cells compared to controls, and insensitive to the SK channel inhibitor apamin. Taken together, these results suggest that RyR2 plays a critical role in regulating PLC activity and PIP2 levels via regulation of SOCE. RyR2 also regulates β-cell electrical activity by controlling Cav current density and SK channel activation.

ObjectiveSome individuals with differences of sex development (DSD) conditions undergo medically indicated prophylactic gonadectomy. Gonads of individuals with DSD can contain germ cells and precursors and patients interested in future fertility preservation and hormonal restoration can participate in DSD-specific research protocols to cryopreserve this tissue. However, it is unclear how many providers or institutions offer gonadal tissue cryopreservation (GTC) and how widespread GTC for DSD is across the United States (US). The Pediatric Initiative Network (PIN) and Non-Oncologic Conditions committees of the Oncofertility Consortium sought to assess the current state of GTC for patients with DSD.MethodsAn electronic survey was sent to providers caring for patients with DSD via special interest groups of professional societies and research networks.ResultsThe survey was administered between November 15, 2021 and March 14, 2022. A total of 155 providers responded to the survey, of which 132 respondents care for patients with DSD, and 78 work at facilities that offer medically indicated gonadectomy to patients with DSD diagnoses. They represented 55 US institutions including 47 pediatric hospitals, and 5 international sites (Canada, Denmark, Germany, Qatar). Of individual providers, 41% offer cryopreservation after prophylactic gonadectomy for patients with DSD (32/78). At an institutional level, GTC after medically indicated gonadectomy is available at 54.4% (24/46) of institutions. GTC is offered for a variety of DSD diagnoses, most commonly 45,X/46,XY DSD (i.e., Turner Syndrome with Y-chromosome material and mixed gonadal dysgenesis), ovotesticular DSD, complete androgen insensitivity syndrome (CAIS), and complete gonadal dysgenesis. Responses demonstrate regional trends in GTC practices with 83.3% of institutions in the Midwest, 66.7% in the Northeast, 54.6% in the West, and 35.3% in the South providing GTC. All represented institutions (100%) send gonadal tissue for pathological evaluation, and 22.7% preserve tissue for research purposes.ConclusionsGTC after gonadectomy is offered at half of the US institutions represented in our survey, though a minority are currently preserving tissue for research purposes. GTC is offered for several DSD conditions. Future research will focus on examining presence and quality of germ cells to support clinical decision making related to fertility preservation for patients with DSD.

Spur cell anemia (SCA) is an acquired form of non-autoimmune hemolytic anemia that occurs in advanced liver disease. It is characterized by the presence of acanthocytes or spur cells, spiculated erythrocytes whose shortened life span causes anemia that is unresponsive to transfusion. SCA has been regarded as a rare condition with an ominous prognosis for which the only known cure is liver transplantation, but recent prospective studies have demonstrated the existence of a milder form of SCA in which there are smaller numbers of acanthocytes, but which is nevertheless associated with hemolysis and poor outcomes. This form of SCA appears to be considerably more common than the severe classical variant. The conventional understanding of the pathogenesis of SCA is that abnormalities of lipid metabolism are the primary event driving the formation of spur cells. However, the studies that underpin this theory are based on small numbers of patients with heterogeneous clinical features and inconsistent use of nomenclature for dysmorphic red blood cells. In this review, we discuss the evolution of the current understanding of SCA and therapeutic strategies that have been employed based on this understanding. Our goal is to raise awareness of this understudied condition that has significant implications for patient outcomes. Furthermore, we highlight the need for rigorous, contemporary research into the underlying cause or causes of SCA in order to develop an effective therapy for this disorder.

Abstract As an essential micronutrient for many organisms, sodium plays an important role in ecological and evolutionary dynamics. Although plants mediate trophic fluxes of sodium, from substrates to higher trophic levels, relatively little comparative research has been published about plant growth and sodium accumulation in response to variation in substrate sodium. Accordingly, we carried out a systematic review of plants' responses to variation in substrate sodium concentrations. We compared biomass and tissue‐sodium accumulation among 107 cultivars or populations (67 species in 20 plant families), broadly expanding beyond the agricultural and model taxa for which several generalizations previously had been made. We hypothesized a priori response models for each population's growth and sodium accumulation as a function of increasing substrate NaCl and used Bayesian Information Criterion to choose the best model. Additionally, using a phylogenetic signal analysis, we tested for phylogenetic patterning of responses across taxa. The influence of substrate sodium on growth differed across taxa, with most populations experiencing detrimental effects at high concentrations. Irrespective of growth responses, tissue sodium concentrations for most taxa increased as sodium concentration in the substrate increased. We found no strong associations between the type of growth response and the type of sodium accumulation response across taxa. Although experiments often fail to test plants across a sufficiently broad range of substrate salinities, non‐crop species tended toward higher sodium tolerance than domesticated species. Moreover, some phylogenetic conservatism was apparent, in that evolutionary history helped predict the distribution of total‐plant growth responses across the phylogeny, but not sodium accumulation responses. Our study reveals that saltier plants in saltier soils proves to be a broadly general pattern for sodium across plant taxa. Regardless of growth responses, sodium accumulation mostly followed an increasing trend as substrate sodium levels increased.

To improve the selective delivery of cisplatin (Cis) to cancer cells, we report and establish the significance of active, targeting drug delivery nanosystems for efficient treatment of lung cancer. Specifically, pH-responsive nano-sized zeolitic imidazolate framework (nZIF-90) was synthesized, post-synthetically modified with an Arg-Gly-Asp peptide motif (RGD@nZIF-90), a known cancer cell homing peptide, and loaded with a large amount of Cis (RGD@Cis⊂nZIF-90). RGD@Cis⊂nZIF-90 was shown to be highly stable under physiological conditions (pH = 7.4) with framework dissociation occurring under slightly acidic conditions (pH = 5.0)–conditions relevant to tumor cells–from which 90% of the encapsulated Cis was released in a sustained manner. In vitro assays demonstrated that RGD@Cis⊂nZIF-90 achieved significantly better cytotoxicity (65% at 6.25 μg ml−1) and selectivity (selectivity index = 4.18 after 48 h of treatment) against adenocarcinoma alveolar epithelial cancer cells (A549) when compared with the unmodified Cis⊂nZIF-90 (22%). Cellular uptake using A549 cells indicated that RGD@Cis⊂nZIF-90 was rapidly internalized leading to significant cell death. After successfully realizing this nanocarrier system, we demonstrated its efficacy in transporting and delivering Cis to cancer cells.

Aim: To determine overall survival (OS) and glycemic control in patients with cancer and diabetes. Materials & methods: Patients of our institution with breast, colon, lung, pancreas and prostate cancer were retrospectively reviewed. OS was compared between matched patients with and without diabetes, and changes in glucose value over time were assessed. Results: For 3934 patients each with and without diabetes, adjusted analysis showed no difference in OS according to diabetes status (hazard ratio: 1.07; 95% CI: 0.96–1.20). Mean glucose values decreased over time in patients with and without diabetes (p = 0.01). Conclusion: In this large study of patients with five common cancers, the co-occurrence of diabetes did not affect OS. Cancer did not adversely affect glucose levels.

GLMN is a gene that encodes a critical protein necessary for normal vascular development. Mutations of GLMN predispose individuals to development of glomangiomas, with nearly 100% penetrance by age 30. Glomangiomas are tumors of the glomus body, a thermoregulatory arterial-venous shunt composed of modified smooth muscle cells. Vulvar glomangioma is an exceedingly rare cause of chronic pelvic pain, that may be easily confused for other conditions such as Bartholin’s gland abscess or deep angiomxyomas, thereby delaying diagnosis and treatment. Glomangiomas have characteristic pathologic and imaging findings which may aid diagnosis. We herein describe the case of a 24-year-old female who developed chronic pelvic pain in the setting of a vulvar glomangioma. We further delineate the magnetic resonance imaging and biopsy findings critical to her diagnosis, and the appropriate steps taken for surgical management. She was found to harbor a heterozygous GLMN mutation. To the best of our knowledge, this is the first description of such a case in the medical literature.

Although immigration enforcement has been linked to poor health outcomes among Latinx populations in the United States, little is known about how interpersonal interactions between immigrants and immigration enforcement agents potentially influence Latinx immigrant health. The author estimated heterogeneity in health outcomes among deported Mexican immigrants by experiences of immigration enforcement abuse using data from respondents to the Survey of Migration in the Northern Border of Mexico ( n = 28,853). Experiences of abuse were significantly associated with worse self-rated health and various health symptoms, with particularly large disparities for mental health symptomology. Given the magnitude of the deportation regime enacted against Mexican immigrants in the United States, thousands of immigrants may return to Mexico at an elevated risk for relatively poorer health. These results emphasize the importance of considering micro-level interactions as a mechanism of health stratification within the larger system of immigration enforcement in the United States.

Why sexual reproduction is so common when asexual reproduction should be much more efficient and less costly remains an open question in evolutionary biology. Comparisons between otherwise similar sexual and asexual taxa allow us to characterize the genetic architecture underlying asexuality, which can, in turn, illuminate how this reproductive mode transition occurred and the mechanisms by which it is maintained or disrupted. Here, we used transcriptome sequencing to compare patterns of ovarian gene expression between actively reproducing obligately sexual and obligately asexual females from multiple lineages of Potamopyrgus antipodarum, a freshwater New Zealand snail characterized by frequent separate transitions to asexuality and coexistence of otherwise similar sexual and asexual lineages. We also used these sequence data to evaluate whether population history accounts for variation in patterns of gene expression. We found that source population was a major source of gene expression variation, and likely more influential than reproductive mode. This outcome for these common garden-raised snails is strikingly similar to earlier results from field-collected snails. While we did not identify a likely set of candidate genes from expression profiles that could plausibly explain how transitions to asexuality occurred, we identified around 1,000 genes with evidence of differential expression between sexual and asexual reproductive modes, and 21 genes that appear to exhibit consistent expression differences between sexuals and asexuals across genetic backgrounds. This second smaller set of genes provides a good starting point for further exploration regarding a potential role in the transition to asexual reproduction. These results mark the first effort to characterize the causes of asexuality in P. antipodarum, demonstrate the apparently high heritability of gene expression patterns in this species, and hint that for P. antipodarum, transitions to asexuality might not necessarily be strongly associated with broad changes in gene expression.

Anthropogenic changes to soil properties and development can dominate soil systems, particularly in coal mining-impacted landscapes of the Appalachian region of the United States. Historical mining operations deposited spoils which are developing into mine soils in chronosequences, allowing for a correlation between emplacement age and rates of change in soil properties. The study site was in the Huff Run Watershed (Mineral City, OH, USA) with a series of eleven spoil piles that were deposited over a 30-year time period. Surface soils were analyzed for bulk density, loss on ignition (LOI) as a proxy for organic matter, particle size, and bulk mineralogical (by X-ray diffraction) and elemental (by X-ray fluorescence) compositions. The following linear trends were observed across the transect from older to younger mine soils: bulk density increased from 1.0 cm−3 to 1.5 g cm−3; LOI decreased from ~20% to 5%; the content of sand-sized particles and quartz decreased from ~50% to 30% and 50% to 25%, respectively, with a corresponding increase in the contribution of clay mineral from ~25% to 60%; and Fe and other trace metals (Cu, Ni, Pb, Sb, Sn, and Te) decreased in concentration, while Al, Mg, and K increased in concentration. These trends are likely the result of: (1) organic matter accumulation as vegetation becomes more abundant over time; (2) transport of clays out of more recently emplaced waste; and (3) oxidative dissolution of primary sulfides releasing Fe and other trace metals followed by re-precipitation of secondary Fe-phases and trace metal sequestration. The findings presented here provide insight into the future behavior of these materials and can potentially be used to assess the inferred age of previously unexamined mine soils across a wider geographic area. These results can also inform decisions related to reclamation activities and ecosystem restoration.

A new species of blind subterranean ground beetle in the genus Anillinus Casey is described from Florida. Anillinus albrittonorum sp. nov. (type locality: 6 miles NW High Springs, Columbia County, Florida) has a unique structure of female genitalia and occupies an isolated position within the genus. This new species is illustrated with images of the habitus, body parts, and male and female genitalia. Relationships of A. albrittonorum to other members of the genus are discussed.

Whether the adult testis harbours a somatic progenitor population is unknown. Here, the authors provide evidence that the testis interstitial cells expressing the transcription factor Tcf21 maintain adult testis homeostasis during aging, and act as potential reserve somatic progenitors following injury.

Objective/Context: This article examines the place of the Chilean consulate in conflicts related to control over labor in western Argentina during the 1860s, a decade of national consolidation and economic expansion. It explores how changes in the laboring classes’ experiences and interests in the mid-nineteenth century expressed themselves through the Chilean consulate and the meaning of being Chilean. Methodology: Using consular records, foreign relations records, and criminal records, it analyzes how Chilean laborers—and Argentines claiming to be Chilean—petitioned the consulate to protect them from military conscription and labor abuses. Originality: While transnational labor migrations feature more prominently in histories of the late-nineteenth and early twentieth centuries than in those on the post-independence period, this article adds to understandings of the end of the post-independence period through such migrations and the institutions that supported them. Conclusions: By focusing on the interaction of laborers and the Chilean consulate, this article makes the case for a correlation between non-European transnational labor migrations and the development of a sense of political belonging distinct from republicanism, patriotism, or civic participation, and one that sought exclusion from the obligations of society and one decidedly more oriented towards notions of class and nationality.