Sample Collection Research Articles

Novel geological framework to understand the origin and diversity of orthopyroxene, olivine, spinel (OOS) lithologies on the Moon

A novel geological framework is presented to address the origin of Orthopyroxene, Olivine, Mg-spinel (OOS) lithologies on the Moon, an emerging component of the lunar crust. The new framework can explain multiple remote sensing observations including the rare occurrence of OOS lithologies in close proximity, lack of mafic mineral association with Mg-spinel lithology and the contrasting observation of mafic mineral association with Mg-spinel in returned lunar samples. We further report new OOS exposures at Thomson crater and present the first remote-sensing-based evidence of mafic mixing among various OOS components. Collectively, the new framework and observations provide a testable set of geological scenarios to understand the origin and diversity of the OOS lithologies and their role in lunar crustal diversity. Future missions would directly benefit from this knowledge towards collecting the best OOS lithology samples, currently absent from the returned lunar sample collection.

A single microfluidic device for multi-omics analysis sample preparation.

Combining different "omics" approaches, such as genomics and proteomics, is necessary to generate a detailed and complete insight into microbiome comprehension. Proper sample collection and processing and accurate analytical methods are crucial in generating reliable data. We previously developed the ChipFilter device for proteomic analysis of microbial samples. We have shown that this device coupled to LC-MS/MS can successfully be used to identify microbial proteins. In the present work, we have developed our workflow to analyze concomitantly proteins and nucleic acids from the same sample. We performed lysis and proteolysis in the device using cultures of E. coli, B. subtilis, and S. cerevisiae. After peptide recovery for LC-MS/MS analysis, DNA from the same samples was recovered and successfully amplified by PCR for the 3 species. This workflow was further extended to a complex microbial mixture of known compositions. Protein analysis was carried out, enabling the identification of more than 5000 proteins. The recovered DNA was sequenced, performing comparable to DNA extracted with a commercial kit without proteolysis. Our results show that the ChipFilter device is suited to prepare samples for parallel proteomic and genomic analyses, which is particularly relevant in the case of low-abundant samples and drastically reduces sampling bias.

Your Blood is Out for Delivery: Considerations of Shipping Time and Temperature on Degradation of RNA from Stabilized Whole Blood.

Remote research studies are an invaluable tool for reaching populations with limited access to large medical centers or universities. To expand the remote study toolkit, we previously developed homeRNA, which allows for at-home self-collection and stabilization of blood and demonstrated the feasibility of using homeRNA in high temperature climates. Here, we expand upon this work through a systematic study exploring the effects of high temperature on RNA integrity (represented as RNA Integrity Number, RIN) through in-lab and field experiments. Compared to the frozen controls (overall mean RIN of 8.2, n = 8), samples kept at 37 °C for 2, 4, and 8 days had mean RINs of 7.6, 5.9, and 5.2 (n = 3), respectively, indicating that typical shipping conditions (∼2 days) yield samples suitable for downstream RNA sequencing. Shorter time intervals (6 h) resulted in minimal RNA degradation (median RIN of 6.4, n = 3) even at higher temperatures (50 °C) compared to the frozen control (mean RIN of 7.8, n = 3). Additionally, we shipped homeRNA-stabilized blood from a single donor to 14 states and back during the summer with continuous temperature probes (7.1 median RIN, n = 42). Samples from all locations were analyzed with 3' mRNA-seq to assess differences in gene counts, with the data suggesting that there was no preferential degradation of transcripts as a result of different shipping times, temperatures, and regions. Overall, our data support that homeRNA can be used in elevated temperature conditions, enabling decentralized sample collection for telemedicine, global health, and clinical research.

The Influence of Performance Expectancy, Effort Expectancy, And Social Influence on Use Behavior with Behavioral Intention as A Mediator (A Case Study of Cash on Delivery (COD) System Users in Marketplaces)

Online shopping has gained significant popularity due to the numerous conveniences it offers. Several marketplaces provide various premium services, including the Cash on Delivery (COD) payment system, which has become a prominent feature in Indonesia. COD caters to individuals who prefer traditional payment methods and are skeptical about online transactions. This payment method allows customers to mitigate concerns about potential fraud, such as non-delivery of goods or receiving empty packages, thereby offering a sense of security and trust in their purchases. This study aims to investigate the relationships between the variables of Performance Expectancy, Effort Expectancy, Social Influence, Behavioral Intention, and Use Behavior. The sample comprises individuals who have shopped on a marketplace using the COD payment system, with a total of 348 respondents. This research employs a quantitative approach, utilizing nonprobability sampling and data collection facilitated by the G-Power application. Data analysis was conducted using Smart-PLS version 4 software. The findings reveal a positive and significant impact of performance expectancy on behavioral intention, a positive and significant effect of effort expectancy on behavioral intention, and a positive and significant influence of social influence on behavioral intention. Furthermore, behavioral intention positively and significantly affects use behavior. Additionally, performance expectancy, effort expectancy, and social influence positively and significantly impact use behavior through behavioral intention.

Assessing heavy metal contamination in agricultural soils: a new GIS-based Probabilistic Pollution Index (PPI) – case study: Guarda Region, Portugal

ABSTRACT Soil contamination by heavy metals is a global agricultural problem, as these elements can be absorbed by plants and transferred to humans through the food pathway. Current contamination assessment methods rely on in situ sample collection, which restricts the evaluation process due to the number of samples that can be analysed and the associated costs. This study addresses the need for a more efficient and cost-effective approach to identifying areas at risk of heavy metal contamination without the logistical constraints of physical sampling. To meet this challenge, we developed a new Probabilistic Pollution Index (PPI), calculated by integrating GIS tools with an 8-parameter probability-risk matrix to identify agricultural areas potentially contaminated by heavy metals. The factors considered included roads, industrial sites, pH levels, soil organic matter content, terrain slope, soil texture, mining areas, and drainage. Each parameter was classified and reclassified to produce a contamination risk map, categorizing each pixel into five levels of risk. To test the PPI, data analysis, parameter classification, and reclassification were applied in the district of Guarda, Portugal. The PPI map revealed that the central portion of the Guarda municipality, along with specific zones in Celorico da Beira, Sabugal, Mêda, and Pinhel, exhibited a high-probability risk of contamination. Given the agricultural expanse within each municipality, enhanced monitoring of heavy metal levels was recommended for Mêda, Pinhel, and Vila Nova de Foz Côa. This index provides a scalable, cost-effective, and easily replicable tool for identifying potential contamination hotspots and the sources and processes of contamination. Designed as a first-line method for large-scale assessments, it supports governmental decision-making and facilitates targeted risk mitigation strategies through a low-cost approach.

Investigator-Initiated Clinical Pharmacokinetic Studies in Resource-Limited Settings: Minimal Requirements and Practical Guidance.

Clinical pharmacology studies are critical for determining the efficacy and safety of drugs. Due to the resource-intensive nature of these studies, most have been conducted in high-income countries, leading to a significant gap in clinical pharmacology data for patients in low- and middle-income countries. This paper provides an overview of the minimal requirements for performing a clinical pharmacology investigator-initiated trial (IIT), including pharmacokinetic sampling. We identify common challenges in resource-limited settings and propose strategies to overcome them. This guideline covers regulatory approval, participant recruitment, drug storage, sample collection and handling, transport, bioanalytical analysis, and data management tailored to the constraints of resource-limited settings. Strategies are proposed to minimize resource demands, including simplified study designs, the use of technologies like whole blood microsampling, and opportunities for collaboration. The goal is to provide practical guidance for those seeking to perform a clinical pharmacology IIT in resource-limited settings to improve safe and effective drug treatment for patients worldwide. Beyond the scope of this guideline is a detailed step-by-step guide on how to perform clinical pharmacology studies.

Addressing the Uncertainties in the Environmental Analysis, Modeling, Source and Risk Assessment of Emerging Contaminants

Emerging contaminants (ECs) have become a growing source of worry for environmental researchers and stakeholders in recent decades. Compared with conventional pollutants, ECs can pose environmental risks even at a trace level. The analysis of ECs is typically significantly more challenging than that of conventional pollutants because of their trace amounts and diverse chemical structures. For sound environmental management, it is necessary to perform a comprehensive study of these pollutants. Global concern has increasingly grown over the occurrence, fate, environmental modeling, and risk assessment of such contaminants. Due to the dearth of knowledge in this area, various unc3ertainties inevitably exist in the investigation of ECs. Environmental problems cannot be precisely understood due to the ubiquitous uncertainties in environmental research. Uncertainties and their sources have been reviewed in this study, including spatial and temporal variability, uncertainty in sample collection and analysis, uncertainty in environmental modeling, uncertainty in risk assessment, and uncertainty in source characterization. Some suggestions to reduce uncertainties are summarized. An awareness of uncertainty is necessary for us to have a more accurate understanding and contribute to sound environmental decision-making and management. In addition, more work remains to be performed to reveal the uncertainties in the analysis and risk assessment of ECs.

Elevated Interleukin-6 Is Associated with an Increased Risk of Long-Term Arteriovenous Fistula Failure for Dialysis

Background/Objectives: The autologous arteriovenous fistula (AVF) is the preferred choice for vascular access in patients with end-stage kidney disease (ESKD) undergoing maintenance hemodialysis. However, in the long term, the primary patency of AVF is suboptimal, with an AVF failure of approximately 30% in one year. The aim of this study is to examine how the pre-operative baseline levels of interleukin-6 (IL-6) affect long-term AVF failure. Methods: This retrospective, observational study involves ESKD patients admitted to the Vascular Surgery Clinic for AVF creation from January 2020 to December 2023. Ultimately, a total of 91 patients whose AVFs matured and began dialysis were enrolled. Prior to surgery, each patient underwent a thorough blood sample collection, with IL-6 levels assessed. The patients were categorized into two groups: those with functioning AVFs and those with dysfunctional AVFs. Their progress was monitored via a review of medical records, telephone interviews, or direct contact. Following the surgery, patients were observed for an average of 1.53 ± 0.94 years. Results: During the follow-up, patients who experienced AVF failure had a higher incidence of diabetes mellitus (p = 0.019) and active smoking (p = 0.012), as well as higher levels of IL-6 (p < 0.001). At ROC analysis, we found a strong association between IL-6 value and AVF failure (AUC: 0.814, p < 0.001), with an optimal cut-off value of 7.08 (76.5% Sensitivity and 79.7% Specificity). Furthermore, at the survival curve Kaplan-Meier analysis, we observed a higher occurrence of AVF failure in patients with baseline IL-6 values above the median (p = 0.004), in tertile 3 (p = 0.002), and above the optimal cut-off value (p < 0.001). At cox-regression analysis, elevated baseline IL-6 levels are associated with AVF Failure (HR: 2.23, p < 0.001). Conclusions: In the current study, we demonstrated that elevated IL-6 levels at baseline are associated with long-term AVF failure, independent of age, sex, and cardiovascular risk factors.

Identification of plant-based spilled oils using direct analysis in real-time–time-of-flight mass spectrometry with hydrophobic paper sampling

Spilled plant-based oils behave very differently in comparison to petroleum oils and require different clean-up measures. They do not evaporate, disperse, dissolve, or emulsify to a significant degree but can polymerize and form an impermeable cap on sediment, smothering benthic media and resulting in an immediate impact on the wildlife community. The current study explored the application of rapid up-to-date direct analysis in real time (DART) with high-resolution mass spectrometry for plant-based oil typing. The study introduced a new concept of using hydrophobic paper to collect and analyze oil samples, thus minimizing sample preparation and expenses. Application of this technique showed its ability to speedily distinguish plant-based from petroleum-based oils. A microcosm experiment exposing plant-based oil samples to weathering processes for comparison with petroleum-based oils demonstrated the ability of the method to classify weathered oil samples and identify their source oil. It was observed that canola and peanut oil were the most resistant to weathering processes. The developed DART-TOFMS method was shown to be accurate for short-term weathered oil spills up to between 12 and 26 days of exposure. The developed method performed identification in less than a day compared to the established multi-day method for oil spill forensics requiring careful sample collection in glass containers, time-consuming laboratory clean-up, lengthy gas chromatography sequences, and careful integration including integration of retention time markers.

Characterization of Renal OAT3 and Hepatic CYP3A Activities in Pregnant Women with Acute Pyelonephritis Using the Endogenous Biomarker Cortisol and 6β-Hydroxycortisol.

This study evaluates the impact of acute pyelonephritis in pregnant women on the in vivo activity of renal OAT3 using the endogenous biomarker (EB) 6β-hydroxycortisol (6β-OHF) renal clearance (CLrenal 6β-OHF) and AUC6β-OHF validated by correlating with the secretion clearance (CLsec) of the probe drug furosemide. Additionally, 6β-OHF formation clearance (CLformation 6β-OHF) as well as urinary (Ae6β-OHF/AeF) and plasma (AUC6βOHF/AUCF) ratios were also evaluated as EB for hepatic CYP3A activity. Pregnant women in their third trimester of gestation, diagnosed with acute pyelonephritis, were recruited before (pre-treatment, n = 8) and after (post-treatment, n = 8) cefuroxime treatment and resolution of acute pyelonephritis. All participants received a single dose of furosemide 40 mg for evaluation of OAT3 in vivo activity on both occasions followed by collection of urine and serial blood samples for 24 h. The CLrenal 6β-OHF (geometric mean and 95% CI) increased from 1.81 L/h (0.86-3.83) to 11.82 L/h (6.58-21.24), whereas the AUC6β-OHF decreased from 44.85 ng h/mL (30.96-64.98) to 24.20 ng h/mL (16.05-36.48) pre- and post-treatment. Significant statistical correlations were observed between furosemide CLsec and CLrenal 6β-OHF (R = 0.88, P =.01) and AUC6β-OHF (R = -0.66, P >.001). Additionally, the CLformation 6β-OHF was lower in pre-treatment 26.81 L/h (10.18-70.59) than in post-treatment 96.18 L/h (64.21-144.09), whereas AUC6βOHF/AUCF ratios were decreased from 0.014 (0.010-0.019) pre-treatment to 0.009 (0.006-0.013) post-treatment. Regarding Ae6β-OHF/AeF ratios, no differences were observed between pre-treatment and post-treatment. In conclusion, CLrenal 6β-OHF evaluates renal OAT3 activity when CYP3A is inhibited, whereas CLformation 6β-OHF evaluates hepatic CYP3A when OAT3 is inhibited, such as in pregnant women with acute pyelonephritis.

Associations Between Daily-Use Products and Urinary Biomarkers of Endocrine-Disrupting Chemicals in Adults of Reproductive Age

Background: Daily-use products, including personal care products, household products, and dietary supplements, often contain ingredients that raise concerns regarding harmful chemical exposure. Endocrine-disrupting chemicals (EDCs) found in daily-use products are associated with numerous adverse health effects. Methods: This pilot study explores the relationship between concentrations of EDCs in urine samples and products used 24 h prior to sample collection, and ingredients of concern in those products, in 140 adults of reproductive age in Northern Nevada. Results: Having higher numbers of products and ingredients of concern, especially in the personal care category, was associated with higher levels of mono-(-ethyl-5-carboxypentyl) phthalate (MECPP). Similarly, taking more supplements was associated with higher levels of methylparaben (MePB). In contrast, using household products with more ingredients of concern was associated with lower levels of monobutyl phthalate (MBP). Generally, women used more products, were exposed to more ingredients of concern and had higher urinary metabolites than men. Participants who rated themselves as being in poor/fair health were exposed to more personal care and supplement ingredients of concern than those in better health. Interestingly, those in excellent health also took supplements with more ingredients of concern. Conclusions: Greater product use and more ingredients of concern are associated with urinary metabolites of known EDCs and self-ratings of poor health. Women and people who take supplements are at greater risk, and even people who consider themselves to be healthy can be highly exposed. More education among the general public is needed to make people aware of the presence of these chemicals in their everyday products so they can make efforts to avoid them.

Novel Method for Routine Ultrasound-Guided Serum Collection for Biomarker Analysis Around the Knee Joint

Background Biomarkers are essential tools in modern medicine, allowing stratification and monitoring of clinical care and treatment response. While systemic blood biomarkers, typically collected from the antecubital vein (AV), are widely used, their sensitivity for joint-specific pathologies such as osteoarthritis (OA) may be limited due to systemic dilution. At present, no serum biomarker reliably reflects the microenvironment of an affected joint in clinical practice. Although synovial fluid (SF) assessment can provide insights into localised pathology and are clinically used for diagnosis in crystal arthropathies and joint infections, their collection is invasive, painful, and carries risks, including infection, making repeated sampling impractical, limiting utility for monitoring treatment responses. This study introduces a novel ultrasound-guided venous sampling technique targeting the saphenous vein (SV) proximal to the knee joint as a less invasive alternative to SF aspiration, hypothesising that it may better reflect the joint-specific microenvironment. Methods A standardised gel model was used to train medically qualified researchers in ultrasound-guided venepuncture of vescles around 2-15 mm in diameter. Subsequently, 32 participants consented to blood sampling from the SV above the knee, with a proximally applied tourniquet to dilate vein diameter for easier venepuncture collection. Results The technique achieved serum collection in over 80% of consented individuals with minimal adverse effects (including n=2 minor bruising and n=1 transient nerve irritation). Key procedural insights included optimal site selection, appropriate pressure application, and effective tourniquet use. Discussion This method demonstrates feasibility, acceptability, and the potential for more localised sample collection, advancing biomarker research. Further validation, including paired SF comparisons, is required to confirm diagnostic utility and develop therapeutic strategies for joint-specific conditions.

Molecular epidemiology and phylogenetic insights of lumpy skin disease in cattle from diverse agro-ecological regions of Punjab, Pakistan.

Lumpy skin disease (LSD) is an emerging, highly contagious transboundary disease of bovines caused by the Lumpy skin disease virus (LSDV), responsible for substantial economic losses to the dairy, meat, and leather industries in Pakistan as well as various countries around the world. Epidemiological information on LSD is scarce in Punjab, Pakistan. Therefore, a molecular epidemiological study was conducted in two agro-ecologically diverse districts (Bhakkar and Jhang) of Punjab, Pakistan. A total of 800 blood samples were randomly collected from the jugular vein of clinically suspected cattle with nodular lesions using a multistage cluster sampling technique. The sampling unit was indigenous, crossbred, and exotic breeds of cattle. Four hundred samples were collected from each district. Ten union councils (UC) were selected from each district, and two villages were selected from each union council. From each village, twenty cattle were selected for sample collection. The PCR-based overall prevalence of LSDV in clinically suspected cattle using the P32 gene was 36.25% (36.25%; 290/800). The multivariable logistic regression analysis indicated that animals who were not treated with acaricide (P = 0.014; OR = 1.459; C.I = 1.079-1.972), body condition score (emaciated animals; P = 0.019; OR = 1.573; CI = 1.076-2.301), and gender (female; (P = 0.016; OR = 1.435; CI = 1.072-1.969) were significantly at higher risk for LSDV infection in cattle. The phylogenetic insights revealed that our isolates were linked to Kenya, China, Russia, Egypt, India, Zimbabwe, Iraq, and Iran. It can be concluded that LSD is widely distributed in the study area, with evidence of genetic diversity. Further studies are required on genetic composition using variable genetic markers for effective control and eradication of LSDV in Pakistan.

Evaluation of Risk Factors Influencing Tick-Borne Infections in Livestock Through Molecular Analyses

Climate changes and human-related activities are identified as major factors responsible for the increasing distribution and abundance of vectors worldwide and, consequently, of vector-borne diseases (VBDs). Farmed animals, during grazing or in establishments with the absence of biosecurity measures, can easily be exposed to wildlife showing high-risk of contagion of several infectious diseases, including VBDs. Furthermore, livestock represents an interface between wildlife and humans, and thus, promoting the transmission pathway of VBDs. Little is known about the presence and prevalence of VBDs in livestock in Southern Italy; therefore, the present study evaluated the circulation of zoonotic VBDs in livestock and potential risk of exposure. A total of 621 whole blood samples belonging to cattle and buffaloes (n = 345) and small ruminants (n = 276) were examined by molecular examinations for the detection of tick-borne pathogens (TBPs). High prevalence (66.3%) for at least one agent was observed. Moreover, the risk of exposure related to environmental features was assessed, as follows: presence of humid areas, high-density of animals, and sample collection during May. These results show a high circulation of TBPs among livestock and underline the need for surveillance in high-risk habitats for public health.

Cardiovascular magnetic resonance-estimated pulmonary capillary wedge pressure, congestion markers, and effect of empagliflozin in patients with heart failure with reduced ejection fraction and dysglycaemia (SUGAR-DM-HF).

A cardiovascular magnetic resonance (CMR) approach to non-invasively estimate left ventricular (LV) filling pressure was recently developed and shown to correlate with invasively measured pulmonary capillary wedge pressure (PCWP). We examined the association between CMR-estimated PCWP (CMR-PCWP) and other imaging and biomarker measures of congestion, and the effect of empagliflozin on these, in the SUGAR-DM-HF trial (NCT03485092). SUGAR-DM-HF enrolled 105 patients with heart failure with reduced ejection fraction (HFrEF) and pre-diabetes or type 2 diabetes who were randomly assigned to empagliflozin 10 mg or placebo once daily for 36 weeks. Transthoracic echocardiography (TTE) was performed at screening and physical examination, lung ultrasound (LUS), CMR, and blood sample collection at baseline and 36 weeks. The mean age of participants was 69 years, mean LV ejection fraction was 33%, and mean CMR-PCWP was 15.1 mmHg (standard deviation 2.5 mmHg). CMR-PCWP correlated with log N-terminal pro-B-type natriuretic peptide (Pearson's r = 0.31, p = 0.0012), total number of B-lines on LUS (Spearman's rho = 0.34, p < 0.001), TTE average E/e' (r = 0.22, p = 0.03) and CMR ventricular volumes (LV end-systolic volume: r = 0.51, p < 0.001; right ventricular end-systolic volume: r = 0.55, p < 0.001). Mean CMR-PCWP decreased from baseline to 36 weeks by -0.37 (95% confidence interval -0.78 to 0.05) mmHg and -0.17 (-0.69 to 0.36) mmHg in the empagliflozin and placebo groups, respectively; placebo-corrected difference -0.43 mmHg (-1.09 to 0.23, p = 0.20). Cardiovascular magnetic resonance-estimated PCWP correlates with clinical and ultrasound markers of congestion in HFrEF. CMR not only enables assessment of cardiac structure and function but also provides a non-invasive assessment of LV filling pressure.

Impact of dietary lysophospholipids supplementation on growth performance, meat quality, and lipid metabolism in finishing bulls fed diets varying in fatty acid saturation

BackgroundThe objective of this study was to evaluate the effects of dietary fatty acids (FA) saturation and lysophospholipids supplementation on growth, meat quality, oxidative stability, FA profiles, and lipid metabolism of finishing beef bulls. Thirty-two Angus bulls (initial body weight: 623 ± 22.6 kg; 21 ± 0.5 months of age) were used. The experiment was a completely randomized block design with a 2 × 2 factorial arrangement of treatments: 2 diets with FA of different degree of unsaturation [high saturated FA diet (HSFA) vs. high unsaturated FA diet (HUFA)] combined with (0.075%, dry matter basis) and without lysophospholipids supplementation. The bulls were fed a high-concentrate diet (forage to concentrate, 15:85) for 104 d including a 14-d adaptation period and a 90-d data and sample collection period.ResultsNo interactions were observed between dietary FA and lysophospholipids supplementation for growth and meat quality parameters. A greater dietary ratio of unsaturated FA (UFA) to saturated FA (SFA) from 1:2 to 1:1 led to lower DM intake and backfat thickness, but did not affect growth performance and other carcass traits. Compared with HSFA, bulls fed HUFA had greater shear force in Longissimus thoracis (LT) muscle, but had lower intramuscular fat (IMF) content and SOD content in LT muscle. Compared with HUFA, feeding the HSFA diet up-regulated expression of ACC, FAS, PPARγ, and SCD1, but down-regulated expression of CPT1B. Compared with feeding HSFA, the HUFA diet led to greater concentrations of c9-C18:1 and other monounsaturated FA in LT muscle. Feeding HUFA also led to lower plasma concentrations of cholesterol, but there were no interactions between FA and lysophospholipids detected. Feeding lysophospholipids improved growth and feed conversion ratio and altered meat quality by increasing muscle pH24h, redness values (24 h), IMF content, and concentrations of C18:3, C20:5 and total polyunsaturated fatty acids. Furthermore, lysophospholipids supplementation led to lower malondialdehyde content and up-regulated the expression of ACC, FAS, and LPL in LT muscle.ConclusionsResults indicated that supplementing a high-concentrate diet with lysophospholipids to beef bulls can enhance growth rate, feed efficiency, meat quality, and beneficial FA. Increasing the dietary ratio of UFA to SFA reduced DM intake and backfat thickness without compromising growth, suggesting potential improvements in feed efficiency.

Gut microbiome dysbiosis is not associated with portal vein thrombosis in patients with end-stage liver disease: a cross-sectional study.

Portal vein thrombosis (PVT) is a common complication in patients with end-stage liver disease (ESLD). The portal vein in ESLD patients is proposedly an inflammatory vascular bed due to translocation of endotoxins and cytokines from the gut. We hypothesized that a pro-inflammatory gut microbiome and elevated trimethylamine N-oxide (TMAO), a driver of thrombosis, may contribute to PVT development. We investigated whether gut microbiome diversity, bacterial species, metabolic pathways, and TMAO levels are associated with PVT in ESLD patients. Fecal samples, plasma samples and data from ESLD patients and healthy controls were collected through the TransplantLines Biobank and Cohort Study. PVT was defined as a thrombus in the portal vein within a year prior to or after fecal sample collection. Fecal samples were analyzed using Shotgun Metagenomic Sequencing, and TMAO levels were measured in plasma using a Vantera® Clinical Analyzer. 102 ESLD patients, of which 23 with PVT, and 246 healthy controls were included. No significant difference in gut microbiome diversity was found between patients with PVT and without PVT (P=0.18). Both ESLD groups had significantly lower alpha-diversity compared with controls. Bacteroides fragilis and three Clostridiales species were increased in patients with PVT compared to without PVT. TMAO levels between the three groups were not significantly different. We observed profound differences in gut microbiota between ESLD patients and controls, but minimal differences between ESLD patients with or without PVT. In our cohort, a gut-derived pro-inflammatory state was not associated with presence of PVT in ESLD patients.

Detection of vaccine-derived poliovirus type 2 from sewage samples and public health response, Poland, November to December 2024.

In October and December 2024, circulating vaccine-derived poliovirus type 2 (cVDPV2) was detected from two wastewater samples in Poland during routine environmental surveillance. The first isolate was characterised and matched previous cVDPV2 isolates detected in Spain in September, as well as in Germany, Finland, and the United Kingdom in November and December 2024. In response to the event, active surveillance for acute flaccid paralysis (AFP) has been strengthened, and the frequency of environmental sample collection has been increased.

The distribution of pigeon adenoviruses in Northern Chinese pigeon and turtledove flocks provides further evidence of viral crosstransmission.

To understand the prevalence, genetic diversity, and potential pathogenicity of adenoviruses present in pigeon and turtledove populations. Nested PCR and Sanger sequencing methods were used to identify the genotype and percentage of various adenoviruses in the feces of pigeon (Columba) and turtledove (Streptopelia) populations. In Beijing, China, a total of 194 fresh feces samples from meat-use pigeons (C livia domestica), homing pigeons (C livia domestica), wild pigeons (C livia domestica), and turtledoves (S decaocto and S chinensis) were collected using noninvasive sampling collection techniques. Their partial DNA-dependent DNA polymerase gene sequences were obtained using nested PCR and double-ended Sanger sequencing, and their genotypes were then ascertained based on sequence alignment. A total of 6 genotypes of adenovirus were detected in pigeon and turtledove flocks, including pigeon adenovirus (PiAdV)-1, PiAdV-2A, PiAdV-3, PiAdV-4, PiAdV-5, and a novel adenovirus genotype (PiAdV-6). Among them, PiAdV-1 was found widespread in flocks of pigeons exhibiting extensive presentations of hepatic necrosis. Highly conserved PiAdV-4 and PiAdV-5 were found to be nonpathogenic and extensively distributed in all pigeon and turtledove groups. These findings imply the presence of diverse PiAdVs in pigeon and turtledove flocks, and the wild pigeons and wild turtledove birds are potentially serving as natural sources of these viruses. This study provides supportive evidence of the pathogenicity of different genotypes of adenovirus in pigeon flocks and also implies that stopping the transmission of the virus brought by wild pigeons and turtledoves may be important for the prevention of diseases associated with PiAdVs.

The compensatory enrichment of sphingosine-1-phosphate on HDL in FSGS enhances the protective function of glomerular endothelial cells compared to MCD

Glomerular endothelial cells (GECs) are pivotal in developing glomerular sclerosis disorders. The advancement of focal segmental glomerulosclerosis (FSGS) is intimately tied to disruptions in lipid metabolism. Sphingosine-1-phosphate (S1P), a molecule transported by high-density lipoproteins (HDL), exhibits protective effects on vascular endothelial cells by upregulating phosphorylated endothelial nitric oxide synthase (p-eNOS) and enhancing nitric oxide (NO) production. Nevertheless, the abundance of S1P within HDL in individuals with FSGS and minimal change disease (MCD) is yet to be elucidated, and its defensive role in GECs necessitates empirical confirmation. A total of 14 FSGS patients, 16 MCD patients, and 16 healthy controls (NC) were included in the study, with FSGS and MCD confirmed by renal biopsy. After blood sample collection, HDL was isolated and categorized into intact HDL, phospholipid-depleted HDL(apo-HDL), phospholipid-remained HDL(phoHDL), and recombinant HDL (rHDL). Various HDL samples, comprising intact, apo-HDL, pho-HDL and rHDL, were co-cultivated with human renal glomerular endothelial cells (HRGECs). Western blotting was utilized to quantify p-eNOS levels and assess PI3K-AKT pathway activation. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyzed S1P concentrations, while real-time quantitative PCR evaluated the expression of enzymes involved in S1P metabolism. Fluorescence labeling methods measured NO levels, and an immunofluorescence colocalization assay investigated Sphingosine-1-phosphate receptor 1 (S1PR1) expression in GECs across distinct kidney tissue groups. The HDL from FSGS patients demonstrated a significantly enhanced ability to promote p-eNOS expression and NO release in HRGECs compared to MCD patients and healthy controls. Additionally, the synthesis activity of S1P in renal tissues of FSGS patients was markedly higher than that observed in MCD patients and healthy controls, suggesting that S1P may play a crucial protective role in the progression of FSGS. Immunofluorescence staining showed that compared with MCD and NC, the expression of S1PR1 in GECs of FSGS patients was significantly decreased. Recombinant HDL with added S1P promoted the increase of p-eNOS in HRGECs. Knockdown of S1PR1 using siRNA reduced the expression of p-eNOS and NO release. The mechanism underlying the regulation of p-eNOS expression by rHDL was associated with the PI3K-AKT signaling pathway. The enhanced presence of S1P on HDL could serve as a diagnostic marker to differentiate FSGS from MCD. Incorporating S1P into HDL enhances glomerular endothelial cell function, suggesting that the S1P/S1PR pathway might offer a promising therapeutic avenue for FSGS.