Application of Proteomics Technology Based on LC-MS Combined with Western Blotting and Co-IP in Antiviral Innate Immunity.

As an interferon-stimulating factor protein, STING plays a role in the response and downstream liaison in antiviral natural immunity. Upon viral invasion, the immediate response of STING protein leads to a series of changes in downstream proteins, which ultimately leads to an antiviral immune response in the form of proinflammatory cytokines and type I interferons, thus triggering an innate immune response, an adaptive immune response in vivo, and long-term protection of the host. In the field of antiviral natural immunity, it is particularly important to rigorously and sequentially probe the dynamic changes in the antiviral natural immunity connector protein STING caused by the entire anti-inflammatory and anti-pathway mechanism and the differences in upstream and downstream proteins. Traditionally, proteomics technology has been validated by detecting proteins in a 2D platform, for which it is difficult to sensitively identify changes in the nature and abundance of target proteins. With the development of mass spectrometry (MS) technology, MS-based proteomics has made important contributions to characterizing the dynamic changes in the natural immune proteome induced by viral infections. MS analytical techniques have several advantages, such as high throughput, rapidity, sensitivity, accuracy, and automation. The most common techniques for detecting complex proteomes are liquid chromatography (LC) and mass spectrometry (MS). LC-MS (Liquid Chromatography-Mass Spectrometry), which combines the physical separation capability of LC and the mass analysis capability of MS, is a powerful technique mainly used for analyzing the proteome of cells, tissues, and body fluids. To explore the combination of traditional proteomics techniques such as Western blotting, Co-IP (co-Immunoprecipitation), and the latest LC-MS methods to probe the anti-inflammatory pathway and the differential changes in upstream and downstream proteins induced by the antiviral natural immune junction protein STING.

Efficacy and safety of combined Chinese and western medicine in the treatment of metabolic syndrome: A network meta-analysis of randomized controlled trials.

Objectives: To comprehensively analyze the randomized controlled clinical trials of integrated traditional Chinese medicine (TCM) and western medicine in the treatment of metabolic syndrome (MetS), and to explore the clinical efficacy and safety of different TCM combined with western medicine for MetS. The purpose of this study is to provide specific suggestions for clinical guidance in the treatment of MetS. Methods: A comprehensive literature review was conducted across several databases, including China Knowledge Network, Wanfang Data, VIP Information, China Biomedical Literature Service System, Embase, PubMed, and Web of Science, up to October 2023. The scope of this review was confined to RCTs focusing on the treatment of metabolic syndrome through an integrated approach of TCM and Western medicine. The primary efficacy endpoints analyzed were clinical efficacy, fasting blood glucose (FBG), triglyceride (TG), and high-density lipoprotein (HDL). Data synthesis and analysis were performed using Stata 16 and RevMan 5.4 for both traditional and network meta-analyses. Results: The findings from both traditional and network meta-analyses reveal that the combination of JiangZhiHuoXue pills (JZHX) + Conventional Western Medicine (CWM) significantly reduces FBG levels. Similarly, the AnShenNingXin capsules (ASNX) + CWM combination markedly lowers TG levels, while the FuFangQiMa capsules (FFQM) + CWM combination shows enhanced efficacy in elevating HDL levels. Notably, the combination of KangNing capsules (KNJN) + CWM demonstrates a more pronounced clinical effect compared to CWM/placebo alone. Conclusions: The study concludes that the synergistic combination of TCM and Western medicine exhibits superior therapeutic benefits in treating MetS compared to CWM/Placebo treatments alone. The combinations of JZHX, AXNX, FFQM, and KNJN with CWM emerge as potentially effective treatments.

A guide to selecting high-performing antibodies for Protein-glutamine gamma-glutamyltransferase 2 (TGM2) for use in western blot, immunoprecipitation and immunofluorescence.

Protein-glutamine gamma-glutamyltransferase 2 (TGM2) is a Ca 2+ dependent enzyme that catalyzes transglutaminase cross-linking modifications. TGM2 is involved in various diseases, either in a protective or contributory manner, making it a crucial protein to study and determine its therapeutic potential. Identifying high-performing TGM2 antibodies would facilitate these investigations. Here we have characterized seventeen TGM2 commercial antibodies for western blot and sixteen for immunoprecipitation, and immunofluorescence. The implemented standardized experimental protocol is based on comparing read-outs in knockout cell lines against their isogenic parental controls. This study is part of a larger, collaborative initiative seeking to address antibody reproducibility issues by characterizing commercially available antibodies for human proteins and publishing the results openly as a resource for the scientific community. While the use of antibodies and protocols vary between laboratories, we encourage readers to use this report as a guide to select the most appropriate antibodies for their specific needs.

Lessons learned: drive-through COVID-19 clinic testing during an adaptive epidemic response and a point-of-care test assessment of a computer-read rapid lateral flow immunoassay with fluorescence-based detection.

Background. The COVID-19 pandemic demonstrated a need for robust SARS-CoV-2 test evaluation infrastructure to underpin biosecurity and protect the population during a pandemic health emergency.Gap statement. The first generation of rapid antigen tests was less accurate than molecular methods due to their inherent sensitivity and specificity shortfalls, compounded by the consequences of self-testing. This created a need for more accurate point-of-care SARS-CoV-2 detection methods.Aim. Here we present the lessons-learned during the COVID-19 emergency response in Western Australia including the detailed set-up, evaluation and operation of rapid antigen test in a state-run drive-through sample collection service during the COVID-19 pandemic after the strict border shutdown ended.Methods. We report a conformity assessment of a novel, second-generation rapid antigen test (Virulizer) comprising a technician-operated rapid lateral flow immunoassay with fluorescence-based detection.Results. The Virulizer rapid antigen test demonstrated up to 100% sensitivity (95% CI: 61.0-100%), 91.94% specificity (95% CI: 82.5-96.5%) and 92.65% accuracy when compared to a commercial PCR assay method. Wide confidence intervals in our series reflect the limits of small sample size. Nevertheless, the Virulizer assay performance was well-suited to point-of-care screening for SARS-CoV-2 in a drive-through clinic setting.Conclusion. The adaptive evaluation process necessary under changing pandemic conditions enabled assessment of a simple sample collection and point-of-care testing process, and showed how this system could be rapidly deployed for SARS-CoV-2 testing, including to regional and remote settings.

Preclinical modeling of metabolic syndrome to study the pleiotropic effects of novel antidiabetic therapy independent of obesity.

Cardiovascular-kidney-metabolic health reflects the interactions between metabolic risk factors, chronic kidney disease, and the cardiovascular system. A growing body of literature suggests that metabolic syndrome (MetS) in individuals of normal weight is associated with a high prevalence of cardiovascular diseases and an increased mortality. The aim of this study was to establish a non-invasive preclinical model of MetS in support of future research focusing on the effects of novel antidiabetic therapies beyond glucose reduction, independent of obesity. Eighteen healthy adult Beagle dogs were fed an isocaloric Western diet (WD) for ten weeks. Biospecimens were collected at baseline (BAS1) and after ten weeks of WD feeding (BAS2) for measurement of blood pressure (BP), serum chemistry, lipoprotein profiling, blood glucose, glucagon, insulin secretion, NT-proBNP, angiotensins, oxidative stress biomarkers, serum, urine, and fecal metabolomics. Differences between BAS1 and BAS2 were analyzed using non-parametric Wilcoxon signed-rank testing. The isocaloric WD model induced significant variations in several markers of MetS, including elevated BP, increased glucose concentrations, and reduced HDL-cholesterol. It also caused an increase in circulating NT-proBNP levels, a decrease in serum bicarbonate, and significant changes in general metabolism, lipids, and biogenic amines. Short-term, isocaloric feeding with a WD in dogs replicated key biological features of MetS while also causing low-grade metabolic acidosis and elevating natriuretic peptides. These findings support the use of the WD canine model for studying the metabolic effects of new antidiabetic therapies independent of obesity.

Molecular prevalence of Dirofilaria sp. hongkongensis, among the dog population in Thiruvananthapuram, India.

Human dirofilariasis is an emerging vector-borne zoonotic parasitic disease in India. Humans are accidental hosts. Symptomatic dirofilariasis, although uncommon is typically manifested in humans as pulmonary, ocular, or subcutaneous nodules. The present research reports the prevalence of the species, Dirofilaria sp. hongkongensis among the dog population in the Kani tribal settlements within the Agasthyamala Biosphere Reserve located at the southernmost end of the Western Ghats, Kerala. To study the prevalence, we conducted a random blood survey among the dog population in these tribal settlements in Thiruvananthapuram. Whole blood was collected from stray and domesticated animals. Genomic DNA was extracted and the microfilariae in the blood were characterized using the slide smear and COI-based marker specific to nematodes. After processing the data from the Sanger sequencer using BLAST, the sequences were submitted to GenBank. Over 25 % of dogs were found positive for Dirofilaria sp. hongkongensis, which is genetically identical to the strain causing human dirofilariasis as is shown by the phylogenetic analysis. The study reveals that the Kani tribes, who reside in deep forests, are particularly vulnerable to this strain of Dirofilaria sp. hongkongensis. Human case reports of this specific strain have been recorded in Kerala over the past decade. The effective elimination is ultimately impacted by a lack of knowledge regarding the mosquito vectors. Dirofilariasis in dogs and humans is on the rise, which demands both active disease surveillance and proper treatment.

Ontology of clinical practice guidelines for Integrated Traditional Chinese and Western Medicine.

OBJECTIVE: Clinical practice guidelines (CPGs) for Integrated Traditional Chinese and Western Medicine (TCM and WM) are important medical documents used to assist medical decision-making and are of great significance for standardizing clinical pathways. However, due to the constraints of text format, it is difficult for Integrated TCM and WM CPGs to play a real role in medical practice. In addition, how to standardize the structure and semantic relationships between Integrated TCM and WM CPG knowledge, and realize the construction of computable, sharable and reliable CPGs, remains an urgent issue to be addressed. Therefore, we are proposing an ontology of CPGs for Integrated TCM and WM. METHODS: We first initialized domain concepts and relationships to ensure the accuracy of the ontology knowledge structure. We then screened CPGs that meet the standards for Integrated TCM and WM, analyzed and classified the contents, and extracted the common structures. Based on the seven-step ontology construction method combined with inference-complement, referring to the representation methods and hierarchical relationships of terms and concepts in MeSH, ICD-10, SNOMED-CT, and other ontologies and terminology sets, we formed the concept structure and semantic relationship tables for the ontology. We also achieved the matching and mapping between the ontology and reference ontologies and term sets. Next, we defined the aspects and constraints of properties, selected multiple Integrated TCM and WM CPGs as instances to populate, and used ontology reasoning tools and formulated defined inference rules to reason and extend the ontology. Finally, we evaluated the performance of the ontology. RESULTS: The content of the Integrated TCM and WM CPGs is divided into nine parts: basic information, background, development method, clinical question, recommendation, evidence, conclusion, result, and reason for recommendations. The Integrated TCM and WM CPG ontology has 152 classes and defines 90 object properties and 114 data properties, with a maximum classification depth of 4 layers. The terms of disease, drug and examination item names in the ontology have been standardized. CONCLUSIONS: This study proposes an Integrated TCM and WM CPG ontology. The ontology adopts a modular design, which has both sharing and scaling ability, and can express rich guideline knowledge. It provides important support for the semantic processing and computational application of guideline documents.

Senescence detection using reflected light.

Senescence is an important cellular program occurring in development, tissue repair, cancer, and aging. Increased senescence is also associated with disease states, including obesity and Type 2 diabetes (T2D). Characterizing and quantifying senescent cells at a single cell level has been challenging and particularly difficult in large primary cells, such as human adipocytes. In this study, we present a novel approach that utilizes reflected light for accurate senescence-associated beta-galactosidase (SABG) staining measurements, which can be integrated with immunofluorescence and is compatible with primary mature adipocytes from both human and mouse, as well as with differentiated 3T3-L1 cells. This technique provides a more comprehensive classification of a cell's senescent state by incorporating multiple criteria, including robust sample-specific pH controls. By leveraging the precision of confocal microscopy to detect X-gal crystals using reflected light, we achieved superior sensitivity over traditional brightfield techniques. This strategy allows for the capture of all X-gal precipitates in SABG-stained samples, revealing diverse X-gal staining patterns and improved detection sensitivity. Additionally, we demonstrate that reflected light outperforms western blot analysis for the detection and quantification of senescence in mature human adipocytes, as it offers a more accurate representation of SABG activity. This detection strategy enables a more thorough investigation of senescent cell characteristics and specifically a deeper look at the relationship between adipocyte senescence and obesity associated disorders, such as T2D.

Methods for Monitoring ER-Phagy.

The endoplasmic reticulum (ER) serves as a central hub for protein synthesis, folding, and lipid biosynthesis in eukaryotic cells. Maintaining ER homeostasis is essential for optimal cellular function, and one mechanism that has garnered attention is endoplasmic reticulum-specific autophagy, or ER-phagy. ER-phagy selectively removes specific ER portions, playing a pivotal role in cellular health and adaptation to environmental stressors. ER-phagy can be induced by diverse cellular conditions such as amino acid starvation, disruption of ER quality control mechanisms, and accumulation of misfolded ER protein, highlighting cellular adaptability and the significance of ER-phagy in stress responses. Clinically relevant mutations in ER-phagy receptors are implicated in various diseases, underlining the fundamental importance of ER-phagy in ER homeostasis. Here, we provide comprehensive protocols and general considerations while investigating ER-phagy using three fundamental techniques-Western blotting, immunofluorescence, and flow cytometry-commonly used in ER-phagy detection and quantitation.

Complete solubilization of mammalian cells in lysates.

In conventional cell lysate protocols, cell debris is typically discarded to obtain a cleaner lysate. However, this approach has limitations, as it may overlook vital cellular components. By discarding cell debris, researchers may inadvertently exclude crucial elements. Retaining all cellular components offers several advantages for studying molecular biology within various cellular compartments. Firstly, it provides a more accurate representation of the cellular environment. Secondly, it enables the study of complex cellular interactions, including those involving cellular structures and signaling pathways associated with debris. This shift in perspective highlights the importance of a holistic approach to lysate preparation. By obtaining lysates that include all cellular components, researchers can gain deeper insights into cellular processes, leading to more accurate data and a better understanding of cellular function and dysfunction. This study aimed to develop a protocol for the preparation of total cell lysates that retain all cellular components, including debris. Our method involves:•A three-step solubilization process using a combination of detergents, saccharides, and chelators, coupled with sonication, in contrast to the classical one-step approach using an all-detergent cocktail.•A comprehensive strategy ensuring the solubilization of all cellular components, providing a more complete lysate for analysis.

The extent of interruptions to primary care medical officers' consultations in the Western Cape.

BACKGROUND:  Administrative tasks are an increasing burden for primary care doctors globally and linked to burnout. Many tasks occur during consultations. They cause interruptions with possible effects on patients' and doctors' experiences and care. The burden and typology of interruptions of doctors in primary care consultations have not been studied in South Africa. Given the link between administrative loads and burnout, describing the extent of these interruptions would help. This study's aim was to assess the extent of interruptions on primary care doctors in the Western Cape. METHODS:  This was a descriptive cross-sectional survey. Doctors from rural and urban primary care clinics in the Western Cape answered an online self-administered survey on the types of interruptions experienced during consultations. Interruptions were categorised and their prevalence calculated. Clinical and non-clinical interruption categories were compared. RESULTS:  There were 201 consultations from 30 doctors. Most interruptions were from retrieving and recording the current patient's information (93.0%), paperwork for other patients (50.7%), and telephone calls about the current patient (41.8%). Other prevalent interruptions were for emergencies (39.8%) and acquiring consumables (37.3%). The median (interquartile range [IQR]) of four (2-4) interruption types per consultation was higher than global settings. CONCLUSION:  Doctors experienced many interruptions during consultations. Their wide range included interruptions unrelated to the current patient.Contribution: This study adds insights from the global south on clinicians' administrative burden. It elaborates on the types of activities that interrupt consultations in an upper-middle income primary care setting. Exploration of interventions to decrease this burden is suggested.

Checklist of marine macroalgae in two contiguous Marine Protected Areas in the south-western Atlantic.

Background: The Costa das Algas Environmental Protection Area (EPA) and the Santa Cruz Wildlife Refuge (WR), located in the Espírito Santo Continental Shelf, Brazil, are outstanding marine protected areas due to their high biodiversity, particularly of macroalgae. Together, these two relatively small protected areas (1,150 and 177 km2, respectively) harbour about a quarter of all macroalgal species recorded in Brazil.The checklist presented herein updates the algal flora of these two protected areas with data obtained until 2019. Two hundred and sixty-five macroalgal taxa were recorded, most of which with vouchers. Checklists based on the collections of each protected area were published on: "Catálogo de Plantas das Unidades de Conservação do Brasil" (https://catalogo-ucs-brasil.jbrj.gov.br/). New information: Besides specimens collected between 2018 and 2019, the algal flora presented herein includes previous records from different Brazilian herbaria (e.g., SP, SPF, ALCB). Herbaria records may include species that do not occur in intertidal reefs (e.g., Laminaria). Overall, 249 macroalgal taxa and one marine angiosperm were recorded in the Costa das Algas EPA (87 new records) and 136 macroalgal taxa and one marine angiosperm in the Santa Cruz WR (46 new records). All taxa are native to Brazil and nine are endemic to Brazil. Our results provide a taxonomic foundation to support management, long-term monitoring and conservation in these protected areas.

The new region demarcation framework for implementing the joint prevention and control of groundwater pollution: A case study in western of Bohai Bay, China.

The joint groundwater pollution prevention and control (GPPC) strategy has been extensively implemented to address the coastal region groundwater pollution challenges in China. However, regional groundwater pollution control and treatment efficiency cannot achieve the expected results due to the lack of regional priority control orders and accurate restoration levels. Thus, this study developed a new region demarcation framework method for delineating GPPC zones, in tandem with a comprehensive pollution index method, the DRASTIC model, source apportionment. To validate the new methodological framework, a case study of groundwater pollution in Qinhuangdao, the western of Bohai Bay, China, was implemented to calculate pollution prevention and control zoning. In total, 340 groundwater samples from shallow aquifers with 9 target pollutants (NO3-, NO2-, NH4+, As, Cd, Cr, Cu, Pb, and Ni) were selected as the dataset for GPPC regionalization. The results showed that GPPC zoning further clarified the direction of groundwater pollution protection and management in Qinhuangdao. Compared to the traditional method, the new GPPC zoning better reflects groundwater mobility characteristics and pollution transport and enrichment patterns in terms of groundwater functional integrity and delineation. This new regional demarcation framework method contributes to providing support for the fine division of groundwater pollution zoning and precise pollution control for groundwater resource management in China.

Systematic review of bovine and zoonotic tuberculosis in the Western Pacific and the Southeast Asia regions of the World Health Organization.

Introduction: Tuberculosis (TB) remains a leading cause of mortality worldwide. We conducted this systematic review to understand the distribution of bovine and zoonotic tuberculosis in the World Health Organization (WHO)'s Southeast Asia Region (SEAR) and Western Pacific Region (WPR) to inform our understanding of the risk posed by this disease. Methods: A two-pronged strategy was used by evaluating data from peer-reviewed literature and official reports. A systematic search was conducted using a structured query in four databases (Web of Science, Scopus, Medline, and PubMed) to identify any reports of the occurrence of zoonotic TB. No language and time constraints were used during the search, but non-English language articles were later excluded. The official data were sourced from the World Organization for Animal Health's (WOAH) World Animal Health Information System (WAHIS) and WHO's global TB database. Results: The retrieved records from SEAR and WPR (n = 113) were screened for eligibility, and data about disease occurrence were extracted and tabulated. In SEAR, all of the five studies that conducted Mycobacterium speciation (5/6) in humans were from India, and the reported Mycobacterium species included M. tuberculosis, M. bovis, M. scrofulacium, M. kansasii, M. phlei, M. smegmatis and M. orygis. In WPR, Mycobacterium speciation investigations in humans were conducted in Australia (8), China (2), Japan (2), NewZealand (2) and Malaysia (1), and the reported Mycobacterium species included M. bovis, M. africanum and M. tuberculosis. Seven countries in WHO's SEAR have officially reported the occurrence of Mycobacterium bovis in their animals: Bangladesh, India, Indonesia, Myanmar, Nepal, Sri Lanka and Thailand. In WPR, the WAHIS information system includes reports of the identification of M. bovis from 11 countries - China, Fiji, Japan, Malaysia, Mongolia, New Zealand, the Philippines, the Republic of Korea, Singapore, Tonga and Viet Nam. In contrast, human zoonotic TB cases in the WHO database were only listed from Australia, Brunei Darussalam and Palau countries. Discussion: The available data suggests under-reporting of zoonotic TB in the regions. Efforts are required to strengthen zoonotic TB surveillance systems from both animal and human health sides to better understand the impact of zoonotic TB in order to take appropriate action to achieve the goal of ending the TB epidemic.

A Brief History of the Progress in Our Understanding of Genetics and Lifestyle, Especially Diet, in the Risk of Alzheimer's Disease.

The two major determining factors for Alzheimer's disease (AD) are genetics and lifestyle. Alleles of the apolipoprotein E (APOE) gene play important roles in the development of late-onset AD, with APOEɛ4 increasing risk, APOEɛ3 being neutral, and APOEɛ2 reducing risk. Several modifiable lifestyle factors have been studied in terms of how they can modify the risk of AD. Among these factors are dietary pattern, nutritional supplements such as omega-3 fatty acids, and B vitamins, physical exercise, and obesity, and vitamin D. The Western diet increases risk of AD, while dietary patterns such as the Mediterranean and vegetarian/vegan diets reduce risk. Foods associated with reduced risk include coffee, fruits and vegetables, whole grains and legumes, and fish, while meat and ultraprocessed foods are associated with increased risk, especially when they lead to obesity. In multi-country ecological studies, the amount of meat in the national diet has the highest correlation with risk of AD. The history of research regarding dietary patterns on risk of AD is emphasized in this review. The risk of AD can be modified starting at least by mid-life. People with greater genetic risk for AD would benefit more by choosing lifestyle factors to reduce and/or delay incidence of AD.

Does kinship with the silverback matter? Intragroup social relationships of immature wild western lowland gorillas after social upheaval.

In primates living in one-male groups, the sole resident male is often an important social partner for group immatures. For such groups, however, replacement of the male and subsequent disruptions of their relationships are almost inevitable. Here, we described social relationships of immature wild western lowland gorillas within a habituated group, where two natal and eight immigrant immatures lived with the resident silverback. We recorded 5 m proximities among group members as an indicator of social closeness. We found that natal immatures spent more time within 5 m of the silverback than immigrant ones. The social closeness between the silverback and the younger immigrant immatures sharply increased after 1 year, but these values were still below those of the natal immatures. Regarding the development of independence from the mother, we found no significant difference between natal and immigrant immatures. The socially preferred nonmother mature for natal immatures was the silverback, whereas many immigrant immatures preferred a paternal adult sister who had previously co-resided with them in a previous group. Our results suggest that familiarity may be an important determinant of the social closeness between the silverback and immatures, but 1 year of co-residence might be too short to construct sufficient familiarity. The paternal sister may have played a pivotal role in the assimilation of immigrant immatures into the non-natal group. Nonetheless, it is not negligible that the silverback and immigrant immatures formed day-to-day close proximities. His tolerance toward co-residence with immigrant immatures can be considered a reproductive tactic.

Palbociclib in HR-Positive, HER2-Negative Advanced/Metastatic Breast Cancer: A Systematic Scoping Review of Real-World Evidence from Countries Outside of Western Regions that Are Underrepresented in Clinical Trials.

INTRODUCTION: Limited awareness exists regarding real-world data (RWD) for palbociclib in patients with hormone receptor-positive, human epidermal growth factor receptor 2-negative advanced/metastatic breast cancer in populations from certain countries outside of Western regions. METHODS: A systematic scoping review was conducted using PubMed and Embase to evaluate RWD for palbociclib from countries outside of Western regions that are underrepresented in clinical trials. Search criteria were aligned with our research question for relevant English-language publications, without restrictions on publication date, followed by Phase 1 (title and abstract) and Phase 2 (full-text) screening of retrieved citations as per Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Data analyses of eligible studies were done separately for abstracts and full-text publications to enhance the precision and reliability of the results. RESULTS: Database search yielded 1485 non-duplicate records, 46 qualified for inclusion, of which 47.8% were published as full text. The analysis of outcomes, based exclusively on full-text publications that collectively included 2048 patients treated with palbociclib, revealed the median progression-free survival (PFS) of 20.2-36.7 months, overall survival (OS) of 39.9 months (reported in one publication) and objective response rate (ORR) of 45.3-80.0% with first-line treatment. In ≥ second line, the median PFS, OS and ORR ranged from 7.0 to 24.2 months, 11 to 19.6 months, and 13.9% to 47.9%, respectively. The safety profile of palbociclib was similar to that reported in pivotal clinical studies, and no new safety concerns were identified. CONCLUSIONS: A comprehensive volume of evidence demonstrates that palbociclib's effectiveness and safety profile in real-world settings align with those observed in clinical trials, offering valuable insights for clinical decision-making in countries outside of Western regions underrepresented in clinical trials.

Revisiting current distribution and future habitat suitability models for the endemic Malabar Tree Toad (Pedostibes tuberculosus) using citizen science data.

Climate change is one of the major drivers of biodiversity loss. Among vertebrates, amphibians are one of the more sensitive groups to climate change due to their unique ecology, habitat requirements, narrow thermal tolerance and relatively limited dispersal abilities. We projected the influence of climate change on an endemic toad, Malabar Tree Toad (Pedostibes tuberculosus; hereafter MTT) from the Western Ghats biodiversity hotspot, India, for two different shared socio-economic pathways (SSP) using multiple modeling approaches for current and future (2061-2080) scenarios. The data used predominantly comes from a citizen science program, 'Mapping Malabar Tree Toad' which is a part of the Frog Watch citizen science program, India Biodiversity Portal. We also evaluated the availability of suitable habitats for the MTT in Protected Areas (PAs) under the current and future scenarios. Our results show that annual precipitation was the most important bioclimatic variable influencing the distribution of MTT. We used MaxEnt (MEM) and Ensemble (ESM) modeling algorithms. The predicted distribution of MTT with selected environmental layers using MEM was 4556.95 km2 while using ESM was 18,563.76 km2. Overlaying PA boundaries on predicted distribution showed 37 PAs with 32.7% (1491.37 km2) and 44 PAs with 21.9% (4066.25 km2) coverage for MEM and ESM respectively. Among eight future climate scenarios, scenarios with high emissions showed a decreased distribution range from 33.5 to 68.7% of predicted distribution in PAs, while scenarios with low emissions showed an increased distribution range from 1.9 to 111.3% in PAs. PAs from the Central Western Ghats lose most suitable areas with a shift of suitable habitats towards the Southern Western Ghats. This suggests that MTT distribution may be restricted in the future and existing PAs may not be sufficient to conserve their habitats. Restricted and discontinuous distribution along with climate change can limit the dispersal and persistence of MTT populations, thus enhanced surveys of MTT habitats within and outside the PAs of the Western Ghats are an important step in safeguarding the persistence of MTT populations. Overall, our results demonstrate the use of citizen science data and its potential in modeling and understanding the geographic distribution and the calling phenology of an elusive, arboreal, and endemic amphibian species.

An enhanced and rapid method for von Willebrand factor multimer analysis for mechanical circulatory device testing.

BACKGROUND: Von Willebrand factor (VWF) is a critical glycoprotein in hemostasis and is an important factor in diagnosing bleeding disorders. Albeit the analysis of VWF is often compromised by inconsistent methodologies and challenges quantifying multimeric size. Current VWF multimer analysis methods are costly, time-consuming, and often inconsistent; thus, demanding skilled professionals. This study aimed to streamline and optimize the VWF multimer analysis technique, making it more efficient and reproducible, particularly for identifying or predicting mechanical circulatory support (MCS) induced bleeding disorders. METHODS: Blood samples from healthy volunteers were exposed to high shear forces via a Medtronic HeartWare ventricular assist device. VWF multimers were analyzed using vertical-gel agarose electrophoresis and Western blotting. Differences in VWF distribution were determined using densitometry, and two methods of densitometric analysis were compared: proprietary software against open-source software. RESULTS: Using the developed method: (i) protocol duration was accelerated from three days (in classical methods) to ~ eight hours; (ii) the resolution of the high molecular weight (HMW) VWF multimers were substantially improved; and (iii) densitometric analysis tools were validated. Additionally, the densitometry analysis using two software types showed a strong correlation between results, with the proprietary software reporting slightly higher HMW VWF percentages. CONCLUSION: This methodology is recommended for affordable, accurate, and reproducible VWF multimer evaluations during MCS use and testing. Further research comparing this method with semi-automated methods would provide additional insight and improve inter-laboratory comparisons.

Vertical Flux of Microplastics in the Deep Subtropical Pacific Ocean: Moored Sediment-Trap Observations within the Kuroshio Extension Recirculation Gyre.

The Kuroshio Extension recirculation gyre in the western North Pacific is an accumulation site of plastic litter transported by the Kuroshio Current. A sediment trap was moored at a depth of 4900 m at Station KEO within the Kuroshio Extension recirculation gyre, and the vertical flux of microplastics in sinking particles of size <1 mm was observed. Forty-one sediment-trap samples collected from July 1, 2014, to October 2, 2016, were analyzed with a micro-Fourier transform infrared spectrometer and microplastics were detected in all samples. Seventeen polymer types were identified, and 90% of the microplastics were less than 100 µm in size. Microplastic sinking was driven by the action of the biological pump, which was in turn driven by seasonal variations in solar radiation and increased surface primary production typical of the spring season. Microplastic mass flux varied from 4.5 × 10-3 to 0.38 mg m-2 day-1 during the sampling period, with a mean and standard deviation of 0.054 ± 0.075 mg m-2 day-1. Extrapolating the annual microplastic mass flux at Station KEO to the entire Kuroshio Extension recirculation gyre, it is estimated that 0.028 million metric tons of microplastics are transported annually to 4900 m depth in this area.