RESUMO
Employee benefits constitute 38.1% of compensation costs, representing a sizeable investment in the workforce. Unlike other forms of support that depend on the actions of individuals throughout the organization, benefits can be changed through decisions at the highest level and influence employees throughout the company. Yet, the literature on benefits has been largely disjointed, resulting in theoretical ambiguity and practical questions about the role of employee benefit experiences in individual employee outcomes. To inform theory and practice, we organized the benefits literature using social exchange theory as a framework and conducted a meta-analysis on the relationships of employee benefit availability, use, and subjective evaluation with perceived organizational support, employee attitudes, and well-being. Our review (k = 134, N = 260,604) found unique relationships between the availability and subjective evaluation of employee benefits and affective organizational commitment, withdrawal intentions, job satisfaction, and well-being, with these relationships partially mediated by perceived organizational support. Benefit use contributed little to these outcomes beyond benefit availability and subjective evaluation. Benefit subjective evaluation was also more strongly related to most outcomes than were benefits availability and use. These relationships varied across types of benefits, with training benefits more strongly related to job satisfaction and health care and retirement benefits more strongly related to turnover intentions. Altogether, this meta-analysis integrates the empirical literature on employee benefits and highlights the implications of benefit experiences and types for the employee-organization relationship and employee well-being. (PsycInfo Database Record (c) 2024 APA, all rights reserved).
RESUMO
We present a novel small molecule antiviral chemotype that was identified by an unconventional cell-free protein synthesis and assembly-based phenotypic screen for modulation of viral capsid assembly. Activity of PAV-431, a representative compound from the series, has been validated against infectious viruses in multiple cell culture models for all six families of viruses causing most respiratory diseases in humans. In animals, this chemotype has been demonstrated efficacious for porcine epidemic diarrhoea virus (a coronavirus) and respiratory syncytial virus (a paramyxovirus). PAV-431 is shown to bind to the protein 14-3-3, a known allosteric modulator. However, it only appears to target the small subset of 14-3-3 which is present in a dynamic multi-protein complex whose components include proteins implicated in viral life cycles and in innate immunity. The composition of this target multi-protein complex appears to be modified upon viral infection and largely restored by PAV-431 treatment. An advanced analog, PAV-104, is shown to be selective for the virally modified target, thereby avoiding host toxicity. Our findings suggest a new paradigm for understanding, and drugging, the host-virus interface, which leads to a new clinical therapeutic strategy for treatment of respiratory viral disease.
Assuntos
Antivirais , Antivirais/farmacologia , Antivirais/química , Humanos , Animais , Proteínas 14-3-3/metabolismo , Complexos Multiproteicos/metabolismo , Interações Hospedeiro-Patógeno/efeitos dos fármacos , Linhagem CelularRESUMO
Cells must sense and respond to sudden maladaptive environmental changes-stresses-to survive and thrive. Across eukaryotes, stresses such as heat shock trigger conserved responses: growth arrest, a specific transcriptional response, and biomolecular condensation of protein and mRNA into structures known as stress granules under severe stress. The composition, formation mechanism, adaptive significance, and even evolutionary conservation of these condensed structures remain enigmatic. Here we provide a remarkable view into stress-triggered condensation, its evolutionary conservation and tuning, and its integration into other well-studied aspects of the stress response. Using three morphologically near-identical budding yeast species adapted to different thermal environments and diverged by up to 100 million years, we show that proteome-scale biomolecular condensation is tuned to species-specific thermal niches, closely tracking corresponding growth and transcriptional responses. In each species, poly(A)-binding protein-a core marker of stress granules-condenses in isolation at species-specific temperatures, with conserved molecular features and conformational changes modulating condensation. From the ecological to the molecular scale, our results reveal previously unappreciated levels of evolutionary selection in the eukaryotic stress response, while establishing a rich, tractable system for further inquiry.
Assuntos
Resposta ao Choque Térmico , Estresse Fisiológico , Resposta ao Choque Térmico/genética , Estresse Fisiológico/genética , Evolução Biológica , Proteínas de Ligação a Poli(A)/genéticaRESUMO
Accurate genotyping of Killer cell Immunoglobulin-like Receptor (KIR) genes plays a pivotal role in enhancing our understanding of innate immune responses, disease correlations, and the advancement of personalized medicine. However, due to the high variability of the KIR region and high level of sequence similarity among different KIR genes, the currently available genotyping methods are unable to accurately infer copy numbers, genotypes and haplotypes of individual KIR genes from next-generation sequencing data. Here we introduce Geny, a new computational tool for precise genotyping of KIR genes. Geny utilizes available KIR haplotype databases and proposes a novel combination of expectation-maximization filtering schemes and integer linear programming-based combinatorial optimization models to resolve ambiguous reads, provide accurate copy number estimation and estimate the haplotype of each copy for the genes within the KIR region. We evaluated Geny on a large set of simulated short-read datasets covering the known validated KIR region assemblies and a set of Illumina short-read samples sequenced from 25 validated samples from the Human Pangenome Reference Consortium collection and showed that it outperforms the existing genotyping tools in terms of accuracy, precision and recall. We envision Geny becoming a valuable resource for understanding immune system response and consequently advancing the field of patient-centric medicine.
RESUMO
The oxygen species on Ag catalysts and reaction mechanisms for ethylene epoxidation and ethylene combustion continue to be debated in the literature despite decades of investigation. Fundamental details of ethylene oxidation by supported Ag/α-Al2O3 catalysts were revealed with the application of high-angle annular dark-field-scanning transmission electron microscopy-energy-dispersive X-ray spectroscopy (HAADF-STEM-EDS), in situ techniques (Raman, UV-vis, X-ray diffraction (XRD), HS-LEIS), chemical probes (C2H4-TPSR and C2H4 + O2-TPSR), and steady-state ethylene oxidation and SSITKA (16O2 â 18O2 switch) studies. The Ag nanoparticles are found to carry a considerable amount of oxygen after the reaction. Density functional theory (DFT) calculations indicate the oxidative reconstructed p(4 × 4)-O-Ag(111) surface is stable relative to metallic Ag(111) under the relevant reaction environment. Multiple configurations of reactive oxygen species are present, and their relevant concentrations depend on treatment conditions. Selective ethylene oxidation to EO proceeds with surface Ag4-O2* species (dioxygen species occupying an oxygen site on a p(4 × 4)-O-Ag(111) surface) only present after strong oxidation of Ag. These experimental findings are strongly supported by the associated DFT calculations. Ethylene epoxidation proceeds via a Langmuir-Hinshelwood mechanism, and ethylene combustion proceeds via combined Langmuir-Hinshelwood (predominant) and Mars-van Krevelen (minor) mechanisms.
RESUMO
Patients with limb loss face the challenge of having an increased risk of skin disease at residual limb sites. Hyperhidrosis is a common concern for persons with amputation and excessive sweat can cause various skin pathology. Recently, microwave thermoablation (MT) was reported as an effective off-label treatment for hyperhidrosis in patients with limb loss. We present a case in which a patient following MT procedure for hyperhidrosis of a transtibial amputation developed multifocal full thickness cutaneous necrosis and deep venous thrombosis. The possible aetiologies of these complications are discussed including vascular congestion and external thermal injury.Such a case warrants the attention of individuals and providers seeking to use MT for off-label purposes, particularly for patients with limb loss, due to the large treatment surface area and potential for temporary or permanent functional loss of the amputated limb.
Assuntos
Hiperidrose , Lesões dos Tecidos Moles , Trombose Venosa , Humanos , Micro-Ondas/efeitos adversos , Extremidade Inferior , Hiperidrose/etiologia , Lesões dos Tecidos Moles/complicações , Trombose Venosa/complicações , Necrose/cirurgia , Necrose/complicações , Resultado do TratamentoRESUMO
We have shown previously that expression of R345W-Fibulin-3 induces epithelial-mesenchymal transition (EMT) in retinal pigment epithelial (RPE) cells. The purpose of the current study was to determine if extracellular vesicles (EVs) derived from RPE cells expressing R345W-Fibulin-3 mutation are sufficient to induce EMT in recipient cells. ARPE-19 cells were infected with luciferase-tagged wild-type (WT)- Fibulin-3 or luciferase-tagged R345W-Fibulin-3 (R345W) using lentiviruses. EVs were isolated from the media by ultracentrifugation or density gradient ultracentrifugation. Transmission electron microscopy and cryogenic electron microscopy were performed to study the morphology of the EVs. The size distribution of EVs were determined by nanoparticle tracking analysis (NTA). EV cargo was analysed using LC-MS/MS based proteomics. EV-associated transforming growth factor beta 1 (TGFß1) protein was measured by enzyme-linked immunosorbent assay. The capacity of EVs to stimulate RPE migration was evaluated by treating recipient cells with WT- or R345W-EVs. The role of EV-bound TGFß was determined by pre-incubation of EVs with a pan-TGFß blocking antibody or IgG control. EM imaging revealed spherical vesicles with two subpopulations of EVs: a group with diameters around 30 nm and a group with diameters over 100 nm, confirmed by NTA analysis. Pathway analysis revealed that members of the sonic hedgehog pathway were less abundant in R345W- EVs, while EMT drivers were enriched. Additionally, R345W-EVs had higher concentrations of TGFß1 compared to control. Critically, treatment with R345W-EVs was sufficient to increase EMT marker expression, as well as cell migration in recipient cells. This EV-increased cell migration was significantly inhibited by pre-incubation of EVs with pan-TGFß-neutralising antibody. In conclusion, the expression of R345W-Fibulin-3 alters the size and cargo of EVs, which are sufficient to enhance the rate of cell migration in a TGFß dependent manner. These results suggest that EV-bound TGFß plays a critical role in the induction of EMT in RPE cells.
Assuntos
Transição Epitelial-Mesenquimal , Vesículas Extracelulares , Cromatografia Líquida , Vesículas Extracelulares/metabolismo , Proteínas Hedgehog/metabolismo , Espectrometria de Massas em Tandem , Células Epiteliais/metabolismo , Fator de Crescimento Transformador beta/metabolismo , Luciferases/metabolismo , Pigmentos da Retina/metabolismoRESUMO
Plastic scintillators, a class of solid-state materials used for radiation detection, were additively manufactured with vat photopolymerization. The photopolymer resins consisted of a primary dopant and a secondary dopant dissolved in a bisphenol A ethoxylate diacrylate-based matrix. The absorptive dopants significantly influence important print parameters, for example, secondary dopants decrease the light penetration depth by a factor > 12 ×. The primary dopant 2,5-diphenyloxazole had minimal impact on the printing process even when loaded at 25 % by mass of the resin. Working curve measurements, which relate energy dose to cure depth, were performed as a function of feature size to further assess the influence of dopants. Photopatterns smaller than 150 µm width had apparent increases in critical energy dose compared to larger photopatterns, while all resins maintained printed features in line gratings with 50 µm of separation. Printed scintillator monoliths were compared to scintillators cast by traditional molding, demonstrating that the layer-by-layer printing process does not decrease scintillation response. A maximum light output of 31 % of a benchmark plastic scintillator (EJ-200) and successful pulse shape discrimination were achieved with 20 % by mass 2,5-diphenyloxazole as the primary dopant and 0.1 % by mass 9,9-dimethyl-2,7-distyrylfluorene as the secondary dopant in printed scintillator samples.
RESUMO
Cells must sense and respond to sudden maladaptive environmental changes-stresses-to survive and thrive. Across eukaryotes, stresses such as heat shock trigger conserved responses: growth arrest, a specific transcriptional response, and biomolecular condensation of protein and mRNA into structures known as stress granules under severe stress. The composition, formation mechanism, adaptive significance, and even evolutionary conservation of these condensed structures remain enigmatic. Here we provide an unprecedented view into stress-triggered condensation, its evolutionary conservation and tuning, and its integration into other well-studied aspects of the stress response. Using three morphologically near-identical budding yeast species adapted to different thermal environments and diverged by up to 100 million years, we show that proteome-scale biomolecular condensation is tuned to species-specific thermal niches, closely tracking corresponding growth and transcriptional responses. In each species, poly(A)-binding protein-a core marker of stress granules-condenses in isolation at species-specific temperatures, with conserved molecular features and conformational changes modulating condensation. From the ecological to the molecular scale, our results reveal previously unappreciated levels of evolutionary selection in the eukaryotic stress response, while establishing a rich, tractable system for further inquiry.
RESUMO
AIMS: To determine if changes in polyamines metabolism occur during non-alcoholic steatohepatitis (NASH) in human patients and mice, as well as to assess systemic and liver-specific effects of spermidine administration into mice suffering from advanced NASH. MATERIALS AND METHODS: Human fecal samples were collected from 50 healthy and 50 NASH patients. For the preclinical studies C57Bl6/N male mice fed GAN or NIH-31 diet for 6 months were ordered from Taconic and liver biopsy was performed. Based on severity of liver fibrosis, body composition and body weight, the mice from both dietary groups were randomized into another two groups: half receiving 3 mM spermidine in drinking water, half normal water for subsequent 12 weeks. Body weight was measured weekly and glucose tolerance and body composition were assessed at the end. Blood and organs were collected during necropsy, and intrahepatic immune cells were isolated for flow cytometry analysis. RESULTS: Metabolomic analysis of human and murine feces confirmed that levels of polyamines decreased along NASH progression. Administration of exogenous spermidine to the mice from both dietary groups did not affect body weight, body composition or adiposity. Moreover, incidence of macroscopic hepatic lesions was higher in NASH mice receiving spermidine. On the other hand, spermidine normalized numbers of Kupffer cells in the livers of mice suffering from NASH, although these beneficial effects did not translate into improved liver steatosis or fibrosis severity. CONCLUSION: Levels of polyamines decrease during NASH in mice and human patients but spermidine administration does not improve advanced NASH.
Assuntos
Hepatopatia Gordurosa não Alcoólica , Humanos , Masculino , Camundongos , Animais , Hepatopatia Gordurosa não Alcoólica/metabolismo , Espermidina/farmacologia , Modelos Animais de Doenças , Poliaminas , Dieta Hiperlipídica , Peso Corporal , Suplementos NutricionaisRESUMO
Understanding the factors that cause endangered populations to either grow or decline is crucial for preserving biodiversity. Conservation efforts often address extrinsic threats, such as environmental degradation and overexploitation, that can limit the recovery of endangered populations. Genetic factors such as inbreeding depression can also affect population dynamics but these effects are rarely measured in the wild and thus often neglected in conservation efforts. Here we show that inbreeding depression strongly influences the population dynamics of an endangered killer whale population, despite genomic signatures of purging of deleterious alleles via natural selection. We find that the 'Southern Residents', which are currently endangered despite nearly 50 years of conservation efforts, exhibit strong inbreeding depression for survival. Our population models suggest that this inbreeding depression limits population growth and predict further decline if the population remains genetically isolated and typical environmental conditions continue. The Southern Residents also had more inferred homozygous deleterious alleles than three other, growing, populations, further suggesting that inbreeding depression affects population fitness. These results demonstrate that inbreeding depression can substantially limit the recovery of endangered populations. Conservation actions focused only on extrinsic threats may therefore fail to account for key intrinsic genetic factors that also limit population growth.
Assuntos
Depressão por Endogamia , Orca , Animais , Endogamia , Orca/genética , Dinâmica Populacional , Seleção GenéticaRESUMO
Quantitative models that simulate the inheritance and evolution of fitness-linked traits offer a method for predicting how environmental or anthropogenic perturbations can affect the dynamics of wild populations. Random mating between individuals within populations is a key assumption of many such models used in conservation and management to predict the impacts of proposed management or conservation actions. However, recent evidence suggests that non-random mating may be underestimated in wild populations and play an important role in diversity-stability relationships. Here we introduce a novel individual-based quantitative genetic model that incorporates assortative mating for reproductive timing, a defining attribute of many aggregate breeding species. We demonstrate the utility of this framework by simulating a generalized salmonid lifecycle, varying input parameters, and comparing model outputs to theoretical expectations for several eco-evolutionary, population dynamic scenarios. Simulations with assortative mating systems resulted in more resilient and productive populations than those that were randomly mating. In accordance with established ecological and evolutionary theory, we also found that decreasing the magnitude of trait correlations, environmental variability, and strength of selection each had a positive effect on population growth. Our model is constructed in a modular framework so that future components can be easily added to address pressing issues such as the effects of supportive breeding, variable age structure, differential selection by sex or age, and fishery interactions on population growth and resilience. With code published in a public Github repository, model outputs may easily be tailored to specific study systems by parameterizing with empirically generated values from long-term ecological monitoring programs.
RESUMO
FDA-approved anti-TNFα biopharmaceuticals are successful in treating a range of autoimmune diseases. However, not all anti-TNFα products are identical in their patient outcomes, suggesting that there may be product-specific differences stemming from protein structural differences, doses and routes of administration. In this work, we focus only on structural and functional differences across three full-length anti-TNFα mAbs (Humira®, Remicade®, and Simponi Aria®) to better understand the implications of such differences on the products' efficacy. For structural characterization, we quantified N-glycans using mass spectrometry and fluorescence labeling. From these studies, we observed that Remicade® had the highest percent of afucosylated glycans (15.5 ± 1.3 %) and the largest number of unique glycans, 28. While Humira® had the fewest unique glycans, 15, and 11.4 ± 0.8 % of afucosylated, high-mannose glycans. For the functional studies we tested TNFα binding via ELISA, FcγRIIIa binding via AlphaLISA and effector function using an ADCC bioreporter assay. Humira® had a significantly lower EC50 (1.9 ± 0.1 pM) for ELISA and IC50 (10.5 ± 1.1 nM) for AlphaLISA, suggesting that Humira® has higher TNFα and FcγRIIIa binding affinity than Remicade® and Simponi Aria®. Humira® was also the most potent in the bioreporter assay with an EC50 value of 0.55 ± 0.03 nM compared to Remicade® (0.64 ± 0.04 nM) and Simponi Aria® (0.67 ± 0.03 nM). This comparison is significant as it highlights functional differences between mAbs with shared mechanisms of action when examined in a single laboratory and under one set of conditions.
Assuntos
Anticorpos Monoclonais , Polissacarídeos , Humanos , Infliximab , Adalimumab/uso terapêutico , Anticorpos Monoclonais/farmacologiaRESUMO
Soft materials that exhibit compliance, programmability, and reconfigurability can have a transformative impact as electronic skin for applications in wearable electronics/soft robotics. There has been significant progress in soft conductive materials; however, achieving electrically controlled and reversible changes in conductivity and circuit connectivity remains challenging. To overcome this limitation, a soft material architecture with reconfigurable conductive networks of silver flakes embedded within a hydrogel matrix is presented. The conductive networks can be reversibly created/disconnected through various stimuli, including current, humidity, or temperature. Such stimuli affect electrical connectivity of the hydrogel by controlling its water content, which can be modulated by evaporation under ambient conditions (passive dehydration), evaporation through electrical Joule heating (active dehydration), or absorption of additional water (rehydration). The resulting change in electrical conductivity is reversible and repeatable, endowing the composite with on-demand reconfigurable conductivity. To highlight this material's unique properties, it is shown that conductive traces can be reconfigured after severe damage and revert to lower conductivity after rehydration. Additionally, a quadruped robot is demonstrated that can respond to stimuli by changing direction following exposure to excess water, thereby achieving reprogrammable locomotion behaviors.
RESUMO
Workers who are exposed to severe situations such as death, harassment, and others' suffering at work are vulnerable to symptoms of posttraumatic stress disorder (PTSD) and severe distress. This distress may extend to their intimate partners, despite their lack of firsthand experience with the traumatic stressors. Although theory and empirical research suggest that employees' traumatic distress can transmit to their partners, the magnitude of these effects and when, how, and why intimate partners develop secondary traumatic symptoms and distress are not as clear. Drawing from crossover theory as an organizing framework (Westman, 2001), our meta-analysis of 276 articles indicates that the relationship between employee PTSD/distress and spouse PTSD/distress is as strong as the relationship between employee trauma exposure and employee PTSD/distress (ρ = .26), suggesting that workers' PTSD/distress is as distressing for partners as the traumatic stressors are for workers encountering them firsthand. Our moderation tests further revealed that the trauma-exposed workers' vulnerability to traumatic stress symptoms was stronger in military than in nonmilitary settings, whereas the extent to which their symptoms crossover to their intimate partners did not vary across occupations. Mediation tests suggest that traumatic stress crossover is partially explained by the worsened quality of the couple's relationship (e.g., increased social support burden and undermining), consistent with the crossover via couple interaction explanation in crossover theory. On the other hand, there was mixed support for the mediating role of the partner's empathy, indicating further research and clarification are needed. Implications for crossover theory and practice are discussed. (PsycInfo Database Record (c) 2023 APA, all rights reserved).
Assuntos
Estresse Ocupacional , Transtornos de Estresse Pós-Traumáticos , Humanos , Transtornos de Estresse Pós-Traumáticos/diagnóstico , Apoio Social , Ansiedade , Local de TrabalhoRESUMO
Applying dynamic equilibrium theory (DET), we examined the temporal dynamics between role overload and three health behaviors (sleep, diet, physical activity). Participants (N = 781) completed five surveys, with 1-month lag between assessments, and the data were analyzed using general cross-lagged panel modeling (GCLM). Results indicated that people had stable health behavior patterns (i.e., there were strong unit effects) that were related to stable role overload patterns (i.e., the chronic role overload and health behavior factors were significantly related). Furthermore, while monthly increases (impulses) in role overload had a negative effect on health behaviors concurrently, health behaviors quickly adapted or regressed back toward previous levels (i.e., there were weak autoregressive and cross-lagged effects after accounting for chronic factors). Impulse response functions were created to show the specific proportion of the initial impulse effect that persisted on each health behavior over time. The results of these response functions indicated that diet and physical activity regressed back to previous levels within 1 month, whereas sleep regressed back to previous levels within 2 months. Collectively, our results suggest that people engage in fairly stable patterns of health behaviors and that these patterns are partly determined by chronic role overload. Our results also suggest that people are generally resilient to temporary changes in role overload, such that the resulting immediate changes in behavior do not persist or become habitual. These results underscore the strength of habits and the resistance to health behavior change, as well as provide support for the use of GCLM for studying DET. (PsycInfo Database Record (c) 2023 APA, all rights reserved).
Assuntos
Exercício Físico , Comportamentos Relacionados com a Saúde , Humanos , Inquéritos e QuestionáriosRESUMO
Many energy technologies that can provide reliable, low-carbon electricity generation are confined to nations that have access to robust technical and economic capabilities, either on their own or through geopolitical alliances. Equally important, these nations maintain a degree of institutional capacity that could lower the risks associated with deploying emergent energy technologies such as advanced nuclear or carbon capture and storage. The complexity, expense, and scrutiny that come with building these facilities make them infeasible choices for most nations. This paradigm is slowly changing, as the pressing need for low-carbon electricity generation and ongoing efforts to develop modular nuclear and carbon capture technologies have opened the door for potentially wider markets, including in nations without substantial institutional capacity. Here, using advanced nuclear technologies as our testbed, we develop new methods to evaluate national readiness for deploying complex energy infrastructure. Specifically, we use Data Envelopment Analysis-a method that eliminates the need for expert judgment-to benchmark performance across nations. We find that approximately 80% of new nuclear deployment occurs in nations that are in the top two quartiles of institutional and economic performance. However, 85% of potential low-carbon electricity demand growth is in nations that are in the bottom two quartiles of performance. We offer iconic paradigms for deploying nuclear power in each of these clusters of nations if the goal is to mitigate risk. Our research helps redouble efforts by industry, regulators, and international development agencies to focus on areas where readiness is low and risk correspondingly higher.
RESUMO
Human immunoglobulin heavy chain (IGH) locus on chromosome 14 includes more than 40 functional copies of the variable gene (IGHV), which are critical for the structure of antibodies that identify and neutralize pathogenic invaders as a part of the adaptive immune system. Because of its highly repetitive sequence composition, the IGH locus has been particularly difficult to assemble or genotype when using standard short-read sequencing technologies. Here, we introduce ImmunoTyper-SR, an algorithmic tool for the genotyping and CNV analysis of the germline IGHV genes on Illumina whole-genome sequencing (WGS) data using a combinatorial optimization formulation that resolves ambiguous read mappings. We have validated ImmunoTyper-SR on 12 individuals, whose IGHV allele composition had been independently validated, as well as concordance between WGS replicates from nine individuals. We then applied ImmunoTyper-SR on 585 COVID patients to investigate the associations between IGHV alleles and anti-type I IFN autoantibodies, which were previously associated with COVID-19 severity.
Assuntos
COVID-19 , Região Variável de Imunoglobulina , Humanos , Região Variável de Imunoglobulina/genética , Genótipo , COVID-19/genética , Sequenciamento de Nucleotídeos em Larga Escala , Cadeias Pesadas de Imunoglobulinas/genética , Autoanticorpos/genéticaRESUMO
The increasing feasibility of assembling large genomic datasets for non-model species presents both opportunities and challenges for applied conservation and management. A popular theme in recent studies is the search for large-effect loci that explain substantial portions of phenotypic variance for a key trait(s). If such loci can be linked to adaptations, 2 important questions arise: 1) Should information from these loci be used to reconfigure conservation units (CUs), even if this conflicts with overall patterns of genetic differentiation? 2) How should this information be used in viability assessments of populations and larger CUs? In this review, we address these questions in the context of recent studies of Chinook salmon and steelhead (anadromous form of rainbow trout) that show strong associations between adult migration timing and specific alleles in one small genomic region. Based on the polygenic paradigm (most traits are controlled by many genes of small effect) and genetic data available at the time showing that early-migrating populations are most closely related to nearby late-migrating populations, adult migration differences in Pacific salmon and steelhead were considered to reflect diversity within CUs rather than separate CUs. Recent data, however, suggest that specific alleles are required for early migration, and that these alleles are lost in populations where conditions do not support early-migrating phenotypes. Contrasting determinations under the US Endangered Species Act and the State of California's equivalent legislation illustrate the complexities of incorporating genomics data into CU configuration decisions. Regardless how CUs are defined, viability assessments should consider that 1) early-migrating phenotypes experience disproportionate risks across large geographic areas, so it becomes important to identify early-migrating populations that can serve as reliable sources for these valuable genetic resources; and 2) genetic architecture, especially the existence of large-effect loci, can affect evolutionary potential and adaptability.
Assuntos
Oncorhynchus mykiss , Salmão , Alelos , Animais , Evolução Biológica , Espécies em Perigo de Extinção , Oncorhynchus mykiss/genética , Salmão/genéticaRESUMO
Post translational modifications (PTMs) have been shown to negatively impact protein efficacy and safety by altering its native conformation, stability, target binding and/or pharmacokinetics. One PTM in particular, shuffled disulfide bonds, has been linked to decreased potency and increased immunogenicity of protein therapeutics. In an effort to gain more insights into the effects of shuffled disulfide bonds on protein therapeutics' safety and efficacy, we designed and further optimized a semi-automated LC-MS/MS method for disulfide bond characterization on two IgG1 protein therapeutics-rituximab and bevacizumab. We also compared originator vs. biosimilar versions of the two therapeutics to determine if there were notable variations in the disulfide shuffling and overall degradation between originator and biosimilar drug products. From our resulting data, we noticed differences in how the two proteins degraded. Bevacizumab had a general upward trend in shuffled disulfide bond levels over the course of a 4-week incubation (0.58 ± 0.08% to 1.46 ± 1.10% for originator) whereas rituximab maintained similar levels throughout the incubation (0.24 ± 0.21% to 0.51 ± 0.11% for originator). When we measured degradation by SEC and SDS-PAGE, we observed trends that correlated with the LC-MS/MS data. Across all methods, we observed that the originator and biosimilar drugs performed similarly. The results from this study will help provide groundwork for comparative disulfide shuffling analysis by LC-MS/MS and standard analytical methodology implementation for the development and regulatory approval of biosimilars.