Interspecies Variation Research Articles

Morphological ant mimics: constrained to imperfection?

Adaptive evolution relies on both heritable variation and selection. Variation is the raw material upon which selection acts, so any mechanism that limits or prevents the generation of heritable variation reduces the power of selection to lead to adaptation. Such limitations are termed evolutionary constraints. While it is widely accepted that constraints play an important role in shaping evolutionary outcomes, their relative importance, as opposed to adaptation, in determining evolutionary outcomes remains a subject of debate. Evolutionary constraints are often evoked as the reason behind the persistence of inaccurate mimicry. Here, we compared the variation and accuracy of body-shape mimicry in ant-mimicking spiders with that of ant-mimicking insects, predicting greater constraints, and hence inaccuracy, in spiders mimicking ants, due to their evolutionary distance from the ant model. We found high inter-species variation in mimetic accuracy, but dorsally, no overall difference in mimetic accuracy between spider and insect mimics, which is inconsistent with a constraint causing inaccurate mimicry. Our study provides empirical evidence suggesting that imperfect mimicry in spiders and insects is predominantly shaped by adaptive processes rather than constraints or chance. Our findings contribute to our understanding of the mechanisms underlying evolutionary diversity and the processes that shape phenotypic outcomes.

Mice, rats, and guinea pigs differ in FMOs expression and tissue concentration of TMAO, a gut bacteria-derived biomarker of cardiovascular and metabolic diseases.

Increased plasma trimethylamine oxide (TMAO) is observed in cardiovascular and metabolic diseases, originating from the gut microbiota product, trimethylamine (TMA), via flavin-containing monooxygenases (FMOs)-dependent oxidation. Numerous studies have investigated the association between plasma TMAO and various pathologies, yet limited knowledge exists regarding tissue concentrations of TMAO, TMAO precursors, and interspecies variability. Chromatography coupled with mass spectrometry was employed to evaluate tissue concentrations of TMAO and its precursors in adult male mice, rats, and guinea pigs. FMO mRNA and protein levels were assessed through PCR and Western blot, respectively. Plasma TMAO levels were similar among the studied species. However, significant differences in tissue concentrations of TMAO were observed between mice, rats, and guinea pigs. The rat renal medulla exhibited the highest TMAO concentration, while the lowest was found in the mouse liver. Mice demonstrated significantly higher plasma TMA concentrations compared to rats and guinea pigs, with the highest TMA concentration found in the mouse renal medulla and the lowest in the rat lungs. FMO5 exhibited the highest expression in mouse liver, while FMO3 was highly expressed in rats. Guinea pigs displayed low expression of FMOs in this tissue. Despite similar plasma TMAO levels, mice, rats, and guinea pigs exhibited significant differences in tissue concentrations of TMA, TMAO, and FMO expression. These interspecies variations should be considered in the design and interpretation of experimental studies. Furthermore, these findings may suggest a diverse importance of the TMAO pathway in the physiology of the evaluated species.

Shining a Light on Symbiosis: N-Fixing Bacteria Boost Legume Growth under Varied Light Conditions

Legumes are a diverse and important group of plants that play a vital role in agriculture, food security, and environmental sustainability. Rhizobia are symbiotic bacteria that form nitrogen-fixing nodules on legume roots, providing the plant with a valuable source of nitrogen. Phenolic acids are a group of secondary metabolites produced by plants that have a wide range of biological functions, including defense against pests and diseases, tolerance to abiotic stresses, and nutrient uptake. In the context of climate change and the imperative for sustainable agriculture, this study delves into the dynamic responses of legume species to varying light intensities and their intricate interactions with soil microorganisms. We investigated the impact of light intensity and rhizobial inoculation on the biomass, nitrate reductase, acid phosphatase, and production of phenolic acids in the roots of four legume species, Trifolium repens, Vicia sativa, Ornithopus compressus, and Coronilla juncea. Plants were grown under three light intensity regimes (low, medium, and high) and inoculated with either rhizobia or a non-inoculated control. The results highlight that shaded-light-adapted species, T. repens and V. sativa, increased root exudate production when exposed to high light intensity. This response aligns with their mining strategy, effectively allocating resources to optimize nutrient acquisition under varying conditions. In contrast, species hailing from well-illuminated environments, O. compressus and C. juncea, displayed distinct strategies by significantly increasing biomass under high irradiance, capitalizing on the available light and nutrients. The mining strategy of legumes emerged as a central theme, influencing biomass production, nitrogen dynamics, and enzymatic activities. The strong correlations between biomass and total nitrogen accumulation underscore the role of the mining strategy in efficient nutrient acquisition. Inoculated plants, which rely more on biological nitrogen fixation (BNF), exhibited lower δ15N values, indicative of a successful mining strategy to acquire and utilize atmospheric nitrogen. Enzymatic activities and phenolic acids exhibited significant interspecies variations, reflecting the adaptability of legumes to different light conditions. The findings of this study could be used to develop new strategies for improving legume stress tolerance, nutrient uptake capacity, and rhizosphere health.

Identification of Virulence Factors in Isolates of Candida haemulonii, Candida albicans and Clavispora lusitaniae with Low Susceptibility and Resistance to Fluconazole and Amphotericin B.

Emerging life-threatening multidrug-resistant (MDR) species such as the C. haemulonii species complex, Clavispora lusitaniae (sin. C. lusitaniae), and other Candida species are considered as an increasing risk for human health in the near future. (1) Background: Many studies have emphasized that the increase in drug resistance can be associated with several virulence factors in Candida and its knowledge is also essential in developing new antifungal strategies. (2) Methods: Hydrophobicity, adherence, biofilm formation, lipase activity, resistance to osmotic stress, and virulence 'in vivo' on G. mellonella larvae were studied in isolates of C. haemulonii, C. albicans, and C. lusitaniae with low susceptibility and resistance to fluconazole and amphotericin B. (3) Results: Intra- and interspecies variability were observed. C. haemulonii showed high hydrophobicity and the ability to adhere to and form biofilm. C. lusitaniae was less hydrophobic, was biofilm-formation-strain-dependent, and did not show lipase activity. Larvae inoculated with C. albicans isolates displayed significantly higher mortality rates than those infected with C. haemulonii and C. lusitaniae. (4) Conclusions: The ability to adhere to and form biofilms associated with their hydrophobic capacity, to adapt to stress, and to infect within an in vivo model, observed in these non-wild-type Candida and Clavispora isolates, shows their marked virulence features. Since factors that define virulence are related to the development of the resistance of these fungi to the few antifungals available for clinical use, differences in the physiology of these cells must be considered to develop new antifungal therapies.

Early detection of Kentucky bluegrass and perennial ryegrass responses to drought stress by measuring chlorophyll fluorescence parameters

AbstractKentucky bluegrass (Poa pratensis L.; drought resistant) and perennial ryegrass (Lolium perenne L.; drought sensitive) are economically important grass species contrasting in drought stress resistance. This study determined the optimal chlorophyll fluorescence parameters to indicate drought incidence and whether the parameters differentiate intra‐ and interspecies variations in drought stress. For each species, nine cultivars were exposed to well‐watered or drought (water withheld) conditions in growth chambers containing high‐throughput photosynthetic imagers to track real‐time responses of maximum quantum efficiency of photosystem II (Fv/Fm), quantum yield of photosystem II (ΦII), non‐photochemical quenching (NPQ), energy‐dependent quenching (qE), and photoinhibition‐associated quenching (qI). Soil moisture content and relative water content of leaf tissues were evaluated. Due to drought, Fv/Fm and ΦII decreased for cultivars of both species but earlier for perennial ryegrass compared to Kentucky bluegrass. The NPQ, qI, and qE values exhibited more dynamic and earlier changes due to drought compared to Fv/Fm and ΦII and allowed for early, mid, and late drought responses to be illustrated. Drought induced an increase in NPQ, qI, and qE values, which corresponded to activation of photoprotection mechanisms, and was exhibited earlier for perennial ryegrass compared to Kentucky bluegrass. As the drought treatment progressed, a decrease or stabilization of low values of NPQ, qI, and qE was observed, and the lowest values were associated with the most drought‐sensitive cultivars. These results indicate important stress tolerance protection mechanisms for grass species and will broadly impact basic and applied grass research as a nondestructive phenotyping tool.

Unraveling the plasticity of translation initiation in prokaryotes: Beyond the invariant Shine-Dalgarno sequence.

Translation initiation in prokaryotes is mainly defined, although not exclusively, by the interaction between the anti-Shine-Dalgarno sequence (antiSD), located at the 3'-terminus of the 16S ribosomal RNA, and a complementary sequence, the ribosome binding site, or Shine-Dalgarno (SD), located upstream of the start codon in prokaryotic mRNAs. The antiSD has a conserved 5'-CCUCC-3' core, but inter-species variations have been found regarding the participation of flanking bases in binding. These variations have been described for certain bacteria and, to a lesser extent, for some archaea. To further analyze these variations, we conducted binding-energy prediction analyses on over 6,400 genomic sequences from both domains. We identified 15 groups of antiSD variants that could be associated with the organisms' phylogenetic origin. Additionally, our findings revealed that certain organisms exhibit variations in the core itself. Importantly, an unaltered core is not necessarily required for the interaction between the 3'-terminus of the rRNA and the region preceding the AUG of the mRNA. In our study, we classified organisms into four distinct categories: i) those possessing a conserved core and demonstrating binding; ii) those with a conserved core but lacking evidence of binding; iii) those exhibiting binding in the absence of a conserved core; and iv) those lacking both a conserved core and evidence of binding. Our results demonstrate the flexibility of organisms in evolving different sequences involved in translation initiation beyond the traditional Shine-Dalgarno sequence. These findings are discussed in terms of the evolution of translation initiation in prokaryotic organisms.

Revisiting the relationship between nitrogen nutrition index and yield across major species

Crop nitrogen (N) fertilization diagnoses via the N nutrition index (NNI)-yield relationship have been tested for several crop species, but a cross-species comparison of that relationship has not been performed yet. This study aimed to perform a cross-species comparison of the relationship between NNI and yield with emphasis on the yield sensitivity to N deficiency, slope of the models. Additionally, we conducted an evaluation to determine the best NNI sampling moment to predict relative yield, with focus on major grain crops. Based on a recently published global dataset to parametrize critical dilution curves, we calculated integrated NNI, instantaneous NNI, relative yield, and relative shoot biomass for annual ryegrass, tall fescue, maize, potato, rice, and wheat. We obtained 238 observations to fit integrated NNI-relative yield linear mixed-effects models and 1606 observations to fit instantaneous NNI-relative yield models. Subsequently, we performed a sensitivity analysis to determine the best NNI sampling moment to predict relative yield, with focus on major grain crops (maize, rice, and wheat). Our results show that there was low inter-species variation of sensitivity to N deficiency, i.e., the slope of the relationship between relative yield and integrated NNI. For grain crops, instantaneous NNI around anthesis demonstrated a better predictive capability for relative yield, outperforming other vegetative stages. This finding contributed to improving the understanding of the association between relative yield and NNI with implications for breeding programs, nutrient management practices, and crop modelling. Most importantly, this study is a contribution to improving the N nutrition diagnosis for several crop species, by using an integral, comparative approach.

Trophic variability of inter- and intra-copepod species in the South Sea of Korea during summer

Spatial variation and interspecies differences in the trophic position (TP) of copepods were investigated using nitrogen isotope ratios of amino acids. In the summer of 2021, coastal waters and the Changjiang diluted water generated clear seawater temperature and salinity fronts in the South Sea of Korea. Paracalanus parvus s. l. was a dominant species in the copepod community, and the second dominant species differed among inshore, intermediate, and offshore sites. The TP of each copepod species was estimated in two ways, considering only metazoan diets (TPGlu, based on glutamic acid and phenylalanine nitrogen isotope ratios) and both metazoan and protistan diets (TPAla, based on alanine and phenylalanine nitrogen isotope ratios). Both TPGlu and TPAla indicated trophic variability among copepod species and the contribution of protistan diets as a food source in the study area. Calanus sinicus showed a similar herbivorous TP of 2.0 in both TPGlu and TPAla, suggesting little contribution from protistan diets. Two copepod species (P. parvus s. l. and Acartia omorii) exhibited TPGlu values of approximately 2.0 but their TPAla values increased from 0.1 to 0.5, indicating mixed diets of both primary producers and protists. The other three copepods (Pseudocalanus sp., Oithona similis, and O. atlantica) showed a wide range in TPGlu (2.4–3.1) and TPAla (2.7–3.4), suggesting that protistan trophic transfers enhance TPAla (by up to 0.5) in omnivorous copepods. We found a spatial variation in the TPs of copepods among water masses by various controlling factors including surface seawater temperature, salinity, and size-fractionated Chl-a. Our findings support that the TP values could be potential indicative of interspecies variability, providing useful information on the composition of the planktonic food web.

Real-time monitoring of a 3D blood-brain barrier model maturation and integrity with a sensorized microfluidic device.

A significant challenge in the treatment of central nervous system (CNS) disorders is represented by the presence of the blood-brain barrier (BBB), a highly selective membrane that regulates molecular transport and restricts the passage of pathogens and therapeutic compounds. Traditional in vivo models are constrained by high costs, lengthy experimental timelines, ethical concerns, and interspecies variations. In vitro models, particularly microfluidic BBB-on-a-chip devices, have been developed to address these limitations. These advanced models aim to more accurately replicate human BBB conditions by incorporating human cells and physiological flow dynamics. In this framework, here we developed an innovative microfluidic system that integrates thin-film electrodes for non-invasive, real-time monitoring of BBB integrity using electrochemical impedance spectroscopy (EIS). EIS measurements showed frequency-dependent impedance changes, indicating BBB integrity and distinguishing well-formed from non-mature barriers. The data from EIS monitoring was confirmed by permeability assays performed with a fluorescence tracer. The model incorporates human endothelial cells in a vessel-like arrangement to mimic the vascular component and three-dimensional cell distribution of human astrocytes and microglia to simulate the parenchymal compartment. By modeling the BBB-on-a-chip with an equivalent circuit, a more accurate trans-endothelial electrical resistance (TEER) value was extracted. The device demonstrated successful BBB formation and maturation, confirmed through live/dead assays, immunofluorescence and permeability assays. Computational fluid dynamics (CFD) simulations confirmed that the device mimics in vivo shear stress conditions. Drug crossing assessment was performed with two chemotherapy drugs: doxorubicin, with a known poor BBB penetration, and temozolomide, conversely a specific drug for CNS disorders and able to cross the BBB, to validate the model predictive capability for drug crossing behavior. The proposed sensorized microfluidic device represents a significant advancement in BBB modeling, offering a versatile platform for CNS drug development, disease modeling, and personalized medicine.

Comparative karyotypic study of fifteen cyprinids (Cyprinidae, Cyprininae) species. An insight into the chromosomal evolution of the tribe Systomini.

The family Cyprinidae is the largest freshwater fish group with 377 genera and over 3,000 described species. However, this group of fish has very limited cytogenetics and advanced molecular cytogenetics information. Therefore, in this study the karyotypes and other chromosomal characteristics of 15 species in the tribe Systomini (Cyprininae) were examined using Ag-NOR staining along with fluorescence in situ hybridization (5S and 18S rDNA). All species share a similar karyotype (2n = 50; NF = 88-100) in both sexes and no differentiated sex chromosome was observed. Chromosomes bearing NOR sites ranged from one to four pairs among the species, mostly mapped adjacent to telomeres in the short arms of distinct pairs in all analyzed species. This difference indicates an extensive rearrangement of chromosomes including genomic differences. The use of the 5S and 18S rDNA probe confirmed the Ag-NOR sites interstitially located in the telomeric regions of distinct chromosomes, characterizing an interspecies variation of these sites. In most of its analyzed species, the signals of 18S rDNA probe corresponded to the Ag-NOR regions, except in Barbonymus altus, B. gonionotus, B. schwanenfeldii and Puntius brevis having these signals on the same as Ag-NOR regions and other sites.

Quantitative pulmonary pharmacokinetics of tetrandrine for SARS-CoV-2 repurposing: a physiologically based pharmacokinetic modeling approach.

Tetrandrine (TET) has been traditionally used in China as a medication to treat silicosis and has recently demonstrated anti-SARS-CoV-2 potential in vitro. By recognizing the disparity between in vitro findings and in vivo performance, we aimed to estimate the free lung concentration of TET using a physiologically based pharmacokinetic (PBPK) model to link in vitro activity with in vivo efficacy. Comparative pharmacokinetic studies of TET were performed in rats and dogs to elucidate the pharmacokinetic mechanisms as well as discern interspecies variations. These insights facilitated the creation of an animal-specific PBPK model, which was subsequently translated to a human model following thorough validation. Following validation of the pharmacokinetic profile from a literature report on single oral dosing of TET in humans, the plasma and lung concentrations were predicted after TET administration at approved dosage levels. Finally, the antiviral efficacy of TET in humans was assessed from the free drug concentration in the lungs. Both in vivo and in vitro experiments thus confirmed that the systemic clearance of TET was primarily through hepatic metabolism. Additionally, the lysosomal capture of basic TET was identified as a pivotal factor in its vast distribution volume and heterogeneous tissue distribution, which could modulate the absorption dynamics of TET in the gastrointestinal tract. Notably, the PBPK-model-based unbound lung concentration of TET (1.67-1.74μg/mL) at the recommended clinical dosage surpassed the in vitro threshold for anti-SARS-CoV-2 activity (EC90 = 1.52μg/mL). Thus, a PBPK model was successfully developed to bridge the in vitro activity and in vivo target exposure of TET to facilitate its repurposing.

Associations with landscape and local‐scale wetland habitat conditions vary among migratory shorebird species during stopovers

Wetlands provide many ecosystem services and functions, including critical stopover habitat for numerous migratory bird species. Yet, loss and degradation of wetlands due to land use and land cover changes have greatly reduced wetland extent worldwide, leading to declines of many migratory shorebirds globally. In the Western Hemisphere, wetlands of the North American Great Plains provide important stopover habitat for shorebirds; however, much remains to be learned about shorebird habitat use during stopovers in this region, including species‐specific associations with landscape‐scale wetland availability and characteristics of individual wetlands. To improve understanding of shorebird habitat use during migration, we conducted shorebird surveys at > 14 000 wetlands throughout Oklahoma, USA, from 2007 to 2009. For 16 shorebird species, we analyzed associations between observed abundance and wetland variables, including within‐wetland characteristics and availability of wetlands at multiple spatial scales surrounding survey locations. We found that the scale at which observed abundance of shorebirds was associated with wetland extent varied among species; 10 species responded most strongly to wetland extent at the 500 m scale, but 5 species and 1 species responded most strongly at 1000 and 2000 m scales, respectively. Observed abundance for most species was generally associated with higher wetland density at broad scales, presence of extensive shallow water, more usable area, and limited vegetation within wetlands, yet there was considerable interspecific variation in the strength and in some cases direction of effects of these within‐wetland characteristics. This interspecies variability highlights the importance of conserving a variety of wetland types with differing characteristics that support the entire shorebird community during migration. By informing optimal habitat requirements for shorebirds during a critical stage of their annual cycle, knowledge gained about species‐specific habitat needs from studies like ours can assist with development of conservation and management strategies that contribute to halting and reversing global shorebird declines.

Early life exposure to perfluorooctanesulfonate (PFOS) impacts vital biological processes in Xenopus laevis: Integrated morphometric and transcriptomic analyses

Perfluorooctanesulfonate (PFOS) is a ubiquitous environmental pollutant associated with increasing health concerns and environmental hazards. Toxicological analyses of PFOS exposure are hampered by large interspecies variations and limited studies on the mechanistic details of PFOS-induced toxicity. We investigated the effects of PFOS exposure on Xenopus laevis embryos based on the reported developmental effects in zebrafish. X. laevis was selected to further our understanding of interspecies variation in response to PFOS, and we built upon previous studies by including transcriptomics and an assessment of ciliogenic effects. Midblastula-stage X. laevis embryos were exposed to PFOS using the frog embryo teratogenesis assay Xenopus (FETAX). Results showed teratogenic effects of PFOS in a time- and dose-dependent manner. The morphological abnormalities of skeleton deformities, a small head, and a miscoiled gut were associated with changes in gene expression evidenced by whole-mount in situ hybridization and transcriptomics. The transcriptomic profile of PFOS-exposed embryos indicated the perturbation in the expression of genes associated with cell death, and downregulation in adenosine triphosphate (ATP) biosynthesis. Moreover, we observed the effects of PFOS exposure on cilia development as a reduction in the number of multiciliated cells and changes in the directionality and velocity of the cilia-driven flow. Collectively, these data broaden the molecular understanding of PFOS-induced developmental effects, whereby ciliary dysfunction and disrupted ATP synthesis are implicated as the probable modes of action of embryotoxicity. Furthermore, our findings present a new challenge to understand the links between PFOS-induced developmental toxicity and vital biological processes.

Pyruvate modulation of redox potential controls mouse sperm motility

Sperm gain fertilization competence in the female reproductive tract through a series of biochemical changes and a requisite switch from linear progressive to hyperactive motility. Despite being essential for fertilization, regulation of sperm energy transduction is poorly understood. This knowledge gap confounds interpretation of interspecies variation and limits progress in optimizing sperm selection for assisted reproduction. Here, we developed a model of mouse sperm bioenergetics using metabolic phenotyping data, quantitative microscopy, and spectral flow cytometry. The results define a mechanism of motility regulation by microenvironmental pyruvate. Rather than being consumed as a mitochondrial fuel source, pyruvate stimulates hyperactivation by repressing lactate oxidation and activating glycolysis in the flagellum through provision of nicotinamide adenine dinucleotide (NAD)+. These findings provide evidence that the transitions in motility requisite for sperm competence are governed by changes in the metabolic microenvironment, highlighting the unexplored potential of using catabolite combination to optimize sperm selection for fertilization.

Gut microbiome play a crucial role in geographical and interspecies variations in mercury accumulation by fish

Mercury (Hg) contamination in fish has raised global concerns for decades. The Hg biotransformation can be manipulated by gut microbiome and it is found to have a substantial impact on the speciation and final fate of Hg in fish. However, the contribution of intestinal microbiota in geographical and interspecies variations in fish Hg levels has not been thoroughly understood. The present study compared the Hg levels in wild marine fish captured from two distinct regions in South China sea. We observed a quite “ironic” phenomenon that MeHg levels in carnivorous fish from a region with minimal human impacts (Xisha Islands, 92 ± 7.2 ng g−1 FW) were much higher than those from a region with severe human impacts (Daya Bay, 19 ± 0.41 ng g−1 FW). Furthermore, the results showed that gut microbiome determined Hg biotransformation and played a crucial role in the variances in fish Hg levels across different geographical locations and species. The intestinal methylators, rather than demethylators, were more significant in affecting Hg biotransformation in fish. The carnivorous species in Xisha Islands exhibited a higher abundance of intestinal methylators, leading to higher MeHg accumulation. Besides, the gut microbiome could be shaped in response to the elevated Hg levels in these fish, which may benefit their adaptation to Hg toxicity and overall health preservation. However, anthropogenic activities (particularly overfishing) in Daya Bay have severely affected the fish population, disrupting the reciprocal relationships between fish and intestinal microbiota and rendering them more susceptible to pathogenic microbes. Overall, this study provided a comprehensive understanding of the role of gut microbiome in Hg bioaccumulation in fish and offered valuable insights into the co-evolutionary dynamics between fish and gut microbiome in the presence of Hg exposure.

Assessing deep convolutional neural network models and their comparative performance for automated medicinal plant identification from leaf images

Medicinal plants have got notable attention in recent years in the field of pharmaceutical and drug research. The high demand of herbal medicine in the rural areas of developing countries and drug industries necessitates correct identification of the medicinal plant species which is challenging in absence of expert taxonomic knowledge. Against this backdrop, we attempted to assess the performance of seven advanced deep learning algorithms in the automated identification of the plants from their leaf images and to suggest the best model from a comparative study of the models. We meticulously trained VGG16, VGG19, DenseNet201, ResNet50V2, Xception, InceptionResNetV2, and InceptionV3 deep neural network models. This training utilized a dataset comprising 5878 images encompassing 30 medicinal species distributed among 20 families. Our approach involved two avenues: the utilization of public data (PI) and a blend of public and field data (PFI), the latter featuring intricate backgrounds. Our study elucidates the robustness of these models in accurately identifying and classifying both interfamily and interspecies variations. Despite variations in accuracy across diverse families and species, the models demonstrated adeptness in these classifications. Comparing the models, we unearthed a crucial insight: the Normalized leverage factor (γω) for DenseNet201 stands at 0.19, elevating it to the pinnacle position for PI with a remarkable 99.64 % accuracy and 98.31 % precision. In the PFI scenario, the same model achieves a γω of 0.15 with a commendable 97 % accuracy. These findings serve as a guiding beacon for shaping future application tools designed to automate medicinal plant identification at the user level.

Examination of Bovine Red Blood Cell Death in Vitro in Response to Pathophysiologic Proapoptotic Stimuli.

Interspecies variations in mammalian red blood cells (RBCs) are observed in circulating RBC lifespan, cell size, fluidity, aggregation, water permeability, metabolism, lipid composition, and the overall proteome. Bovine RBC cell membrane is deficient in phosphatidylcholine and exhibits anomalies in the arrangement of phosphatidylethanolamine within the lipid bilayer. However, like human RBCs, virtually all the aminophospholipid phosphatidylserine (PS) is found within the cytoplasmic side of the cell membrane of intact circulating bovine RBCs. During apoptotic cell death of human and murine RBCs, PS translocates to the outer leaflet of the cell membrane via Ca2+-dependent and -independent signaling mechanisms. However, little is known about this process in bovine RBCs. Using cytofluorometry analyses, we characterized and compared the cell death responses in bovine and human RBCs in vitro exposed to various pathophysiologic cell stressors. Ionic stress, by ionophore treatment, and oxidative stress enhanced cytoplasmic Ca2+ levels and cell membrane PS expression in both bovine and human RBCs. Fever-grade hyperthermia and energy starvation promoted Ca2+ influx and elevated reactive oxygen species levels in both human and bovine RBCs. However, bovine RBCs displayed minimal increases in PS expression elicited by hyperthermia, energy starvation, and extracellular hypertonicity as compared to human RBCs. In response to decreased extracellular osmolality, bovine RBCs exhibited significantly enhanced fragility as compared to human RBCs. Bovine RBCs display differential cell death patterns as compared to human RBCs, only partly explained by increased Ca2+ influx and oxidative stress. Premature removal of circulating RBCs could potentially contribute to the pathogenesis of anemia in cattle caused by a wide range of factors such as systemic diseases, parasitic infections, and nutritional deficiencies.

Multivariate analysis of FcR-mediated NK cell functions identifies unique clustering among humans and rhesus macaques.

Rhesus macaques (RMs) are a common pre-clinical model used to test HIV vaccine efficacy and passive immunization strategies. Yet, it remains unclear to what extent the Fc-Fc receptor (FcR) interactions impacting antiviral activities of antibodies in RMs recapitulate those in humans. Here, we evaluated the FcR-related functionality of natural killer cells (NKs) from peripheral blood of uninfected humans and RMs to identify intra- and inter-species variation. NKs were screened for FcγRIIIa (human) and FcγRIII (RM) genotypes (FcγRIII(a)), receptor signaling, and antibody-dependent cellular cytotoxicity (ADCC), the latter mediated by a cocktail of monoclonal IgG1 antibodies with human or RM Fc. FcγRIII(a) genetic polymorphisms alone did not explain differences in NK effector functionality in either species cohort. Using the same parameters, hierarchical clustering separated each species into two clusters. Importantly, in principal components analyses, ADCC magnitude, NK contribution to ADCC, FcγRIII(a) cell-surface expression, and frequency of phosphorylated CD3ζ NK cells all contributed similarly to the first principal component within each species, demonstrating the importance of measuring multiple facets of NK cell function. Although ADCC potency was similar between species, we detected significant differences in frequencies of NK cells and pCD3ζ+ cells, level of cell-surface FcγRIII(a) expression, and NK-mediated ADCC (P<0.001), indicating that a combination of Fc-FcR parameters contribute to overall inter-species functional differences. These data strongly support the importance of multi-parameter analyses of Fc-FcR NK-mediated functions when evaluating efficacy of passive and active immunizations in pre- and clinical trials and identifying correlates of protection. The results also suggest that pre-screening animals for multiple FcR-mediated NK function would ensure even distribution of animals among treatment groups in future preclinical trials.

Corticosterone and immune responses to dehydration in squamate reptiles.

Many environments present some degree of seasonal water limitations; organisms that live in such environments must be adapted to survive periods without permanent water access. Often this involves the ability to tolerate dehydration, which can have adverse physiological effects and is typically considered a physiological stressor. While having many functions, the hormone corticosterone (CORT) is often released in response to stressors, yet increasing plasma CORT while dehydrated could be considered maladaptive, especially for species that experience predictable bouts of dehydration and have related coping mechanisms. Elevating CORT could reduce immunocompetence and have other negative physiological effects. Thus, such species likely have CORT and immune responses adapted to experiencing seasonal droughts. We evaluated how dehydration affects CORT and immune function in eight squamate species that naturally experience varied water limitation. We tested whether hydric state affected plasma CORT concentrations and aspects of immunocompetence (lysis, agglutination, bacterial killing ability and white blood cell counts) differently among species based on how seasonally water limited they are and whether this is constrained by phylogeny. The species represented four familial pairs, with one species of each pair inhabiting environments with frequent access to water and one naturally experiencing extended periods (>30 days) with no access to standing water. The effects of dehydration on CORT and immunity varied among species. Increases in CORT were generally not associated with reduced immunocompetence, indicating CORT and immunity might be decoupled in some species. Interspecies variations in responses to dehydration were more clearly grouped by phylogeny than by habitat type.

The Effect of Deuteration and Homologation of the Lactam Ring of Nirmatrelvir on Its Biochemical Properties and Oxidative Metabolism.

This study explores the relationship between structural alterations of nirmatrelvir, such as homologation and deuteration, and metabolic stability of newly synthesized derivatives. We developed a reliable synthetic protocol toward dideutero-nirmatrelvir and its homologated analogues with high isotopic incorporation. Deuteration of the primary metabolic site of nirmatrelvir provides a 3-fold improvement of its human microsomal stability but is accompanied by an increased metabolism rate at secondary sites. Homologation of the lactam ring allows the capping group modification to decrease and delocalize the molecule's lipophilicity, reducing the metabolic rate at secondary sites. The effect of deuteration was less pronounced for the 6-membered lactam than for its 5-membered analogue in human microsomes, but the trend is reversed in the case of mouse microsomes. X-ray data revealed that the homologation of the lactam ring favors the orientation of the drug's nitrile warhead for interaction with the catalytic sulfur of the SARS-CoV-2 Mpro, improving its binding. Comparable potency against SARS-CoV-2 Mpro from several variants of concern and selectivity over human cysteine proteases cathepsin B, L, and S was observed for the novel deuterated/homologated derivative and nirmatrelvir. Synthesized compounds displayed a large interspecies variability in hamster, rat, and human hepatocyte stability assays. Overall, we aimed to apply a rational approach in changing the physicochemical properties of the drug to refine its biochemical and biological parameters.