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During the gestation and lactation period, the energy demand in pregnant and lactating bitches is elevated. Non‐esterified fatty acids (NEFAs) are utilized either directly from the fed diet or from body fat storage. High NEFA concentration in the blood plasma leads to an increased risk for diseases. Therefore, measuring blood NEFA concentrations may be an indicator for a period of scarcity. The aim of this study is to explore if serum NEFA concentrations in healthy bitches change during gestation and lactation. Healthy pregnant and lactating bitches were sampled on three appointed dates around parturition. NEFA values were examined with a multiparameter clinical chemistry analyser. All statistical analyses were performed using R. Overall, 38 bitches were enrolled in the study. Twenty‐one bitches were sampled on all three appointed dates. The median NEFA concentration antepartum was 0.73 mmol/L (IQR: 0.59, 1.01); during peak lactation, it was 0.57 mmol/L (IQR: 0.44, 0.82); and around weaning, it was 0.58 mmol/L (IQR: 0.46, 0.73). NEFA concentrations rose slightly with litter size in late gestation. Body condition score had no influence on observed NEFA values. We conclude that NEFA concentrations widely remain within reference ranges in well‐fed pregnant and lactating bitches. Nevertheless, they may be a valuable parameter to assess the actual metabolic status of malnourished pregnant and lactating bitches. [ABSTRACT FROM AUTHOR]

Background and objective Isoliquiritigenin (ISL) is an important pharmacological constituent of Glycyrrhiza glabra, which possesses a range of physiological and pharmacological activities, as well as significant antitumor activity, and can be used as a potential drug for targeted cancer therapy. LINC01503 is an oncogene, which has been closely associated with the malignant biological processes of many cancers. The aim of this study was to investigate the effects of ISL on the proliferation, apoptosis, invasion and migration of lung squamous carcinoma cells by regulating LINC01503. Methods Plasma was collected from lung squamous carcinoma patients and healthy individuals treated at Tangshan People's Hospital from January 2021 to December 2022. The expression of LINC01503 in lung squamous carcinoma plasma, tissues and cells was detected by real-time quantitative fluorescence polymerase chain reaction (qRT-PCR). Lung squamous carcinoma cells were treated with different concentrations of ISL for 24 h, and LINC01503 expression was detected by qRT-PCR. The cells were treated in groups: si-NC group, si-LINC01503 group, DMSO (0.1% dimethyl sulfone) group, ISL group, pc DNA3.1(+)-NC group, pc DNA3.1(+)-LINC01503 group, ISL+pc DNA3.1(+)-NC group and ISL+pc DNA3.1(+)- LINC01503 groups. CCK-8 assay, clone formation assay, flow cytometry, Transwell assay and scratch assay were used to explore the effect of LINC01503 on the functional phenotype of lung squamous carcinoma cells. Results Fluorescence in situ hybridization results showed that the average fluorescence intensity of LINC01503 in tissue microarrays of lung squamous carcinoma patients was higher than that in paracancerous tissues (P<0.05). The expression of LINC01503 in the plasma of patients with lung squamous carcinoma was higher than that in the plasma of healthy individuals (P<0.05). Knockdown of LINC01503 inhibited the proliferation, invasion and migration of lung squamous carcinoma cells and promoted apoptosis (P<0.05). ISL inhibited the proliferation, invasion, migration and promoted apoptosis of lung squamous carcinoma cells (P<0.05). Overexpression of LINC01503 followed by intervention with ISL reversed the promotional effect of overexpression of LINC01503 on the proliferation, invasion and migration of lung squamous carcinoma cells as well as the inhibitory effect on apoptosis (P<0.05). Conclusion LINC01503 was highly expressed in lung squamous carcinoma, and LINC01503 could promote the proliferation, invasion and migration of lung squamous carcinoma cells and inhibit the apoptosis, ISL could inhibit the proliferation, invasion and migration of lung squamous carcinoma cells and promote apoptosis of lung squamous carcinoma cells by regulating the expression of LINC01503. [ABSTRACT FROM AUTHOR]

Myristoylation, the N-terminal addition of the fatty acid myristate to proteins, regulates membrane-bound signal transduction pathways important in cancer cell biology. This modification is catalyzed by two N-myristoyltransferases, NMT1 and NMT2. Zelenirstat is a first-in-class potent oral small molecule inhibitor of both NMT1 and NMT2 proteins. Patients with advanced solid tumors and relapsed/refractory (R/R) B-cell lymphomas were enrolled in an open label, phase I dose escalation trial of oral daily zelenirstat, administered in 28-day cycles until progression or unacceptable toxicity. The endpoints were to evaluate dose-limiting toxicities (DLT) to establish a maximum tolerated dose (MTD), pharmacokinetic parameters, and anticancer activity. Twenty-nine patients were enrolled (25 advanced solid tumor; 4 R/R B-cell lymphoma) and 24 were DLT-evaluable. Dosing ranged from 20 mg once daily (OD) to 210 mg OD without DLT, but gastrointestinal DLTS were seen in the 280 mg cohort. MTD and recommended phase 2 dose were 210 mg OD. Common adverse events were predominantly Gr ≤ 2 nausea, vomiting, diarrhea, and fatigue. Plasma concentrations peaked at 2 h with terminal half-lives averaging 10 h. Steady state was achieved by day 15, and higher doses achieved trough concentrations predicted to be therapeutic. Stable disease as best response was seen in eight (28%) patients. Progression-free survival and overall survival were significantly better in patients receiving 210 mg OD compared to those receiving lower doses. Zelenirstat is well-tolerated, achieves plasma exposures expected for efficacy, and shows early signs of anticancer activity. Further clinical development of zelenirstat is warranted. [ABSTRACT FROM AUTHOR]

Background/objectives: Prolonged fasting triggers a stress response within the human body. Our objective was to investigate the impact of prolonged fasting, in conjunction with stress, on kynurenine pathway metabolites. Subjects/methods: Healthy males were divided into fasting group (zero-calorie-restriction) for 6 days (FAST, n = 14), and control group (CON, n = 10). Blood and saliva samples were collected at baseline, Day 2, Day 4, Day 6 during fasting period, and 1 week after resuming regular diet. Plasma levels of kynurenine pathway metabolites were measured using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS/MS). Plasma and salivary samples were analyzed for stress markers. Results: A pronounced activation of the kynurenine pathway in individuals on FAST trial was revealed. Concentrations of picolinic acid (PIC), kynurenic acid (KYNA) and 3-hydroxykynurenine (3-HK) were significantly increased, with peak levels observed on Day 6 (P < 0.0001). Conversely, concentrations of tryptophan (TRP) and quinolinic acid (QUIN) decreased (P < 0.0001), while kynurenine (KYN) and nicotinamide (NAM) levels remained stable. Cortisol and noradrenaline concentrations remained unchanged. However, adrenaline levels significantly increased on Day 4 within FAST compared to CON (P = 0.005). Notably, all deviations in kynurenine pathway metabolite levels returned to baseline values upon resuming regular diet following the 6-day fasting regimen, even when weight and BMI parameters were not restored. Conclusions: Extended fasting over 6 days induces the kynurenine pathway and has minimal effects on stress markers. Restoration of metabolite concentrations upon regular feeding implies rapid adaptation of the kynurenine pathway synthetic enzymes to maintain homeostasis when faced with perturbations. [ABSTRACT FROM AUTHOR]

Dilated cardiomyopathy (DCM) is one of the major causes of heart failure. Although significant progress has been made in elucidating the underlying mechanisms, further investigation is required for clarifying molecular diagnostic and therapeutic targets. In this study, we found that the mRNA level of protein phosphatase 2 regulatory subunit B' delta (Ppp2r5d) was altered in the peripheral blood plasma of DCM patients. Knockdown of Ppp2r5d in murine cardiomyocytes increased the intracellular levels of reactive oxygen species (ROS) and inhibited adenosine triphosphate (ATP) synthesis. In vivo knockdown of Ppp2r5d in an isoproterenol (ISO)-induced DCM mouse model aggravated the pathogenesis and ultimately led to heart failure. Mechanistically, Ppp2r5d-deficient cardiomyocytes showed an increase in phosphorylation of STAT3 at Y705 and a decrease in phosphorylation of STAT3 at S727. The elevated levels of phosphorylation at Y705 in STAT3 triggered the upregulation of interleukin 6 (IL6) expression. Moreover, the decreased phosphorylation at S727 in STAT3 disrupted mitochondrial electron transport chain function and dysregulated ATP synthesis and ROS levels. These results hereby reveal a novel role for Ppp2r5d in modulating STAT3 pathway in DCM, suggesting it as a potential target for the therapy of the disease. [ABSTRACT FROM AUTHOR]

The study aimed to investigate the impact of wet feeding on broiler chicken performance and intestinal histomorphometry. The research was conducted at the Veterinary College, Udaipur, India, using 144 commercial broiler chicks from day 1 to 6 weeks of age. The chicks were divided randomly into three experimental groups, each with four replicates comprising 48 birds. The control group (G1) was fed dry; while, group 2 (G2) received wet feed supplemented with 1.00 g water per g of feed and group 3 (G3) received wet feed supplemented with 1.20 g water per g of feed. Results revealed that G3 had significantly superior feed conversion ratios, lower feed intake, and increased body weight and weight gain compared to G2 and G1. Among the three treatment groups, there were no variations in blood plasma indicators. In comparison with G1, duodenum and ileum in G2 had higher villus height. The G3 revealed higher net profit margins and 18.89% saving of feed compared to G1, suggesting that it may be a valuable tool for lowering chicken feed costs. [ABSTRACT FROM AUTHOR]

Oxidative stress (OS) could be a condition underlying several human diseases, despite the physiological role of reactive oxygen species (oxidative eustress). Therefore, antioxidant compounds could represent a modulatory mechanism for maintaining a proper redox balance and redox signaling. When antioxidants are insufficient or overwhelmed, OS ensues, causing multiple damages at molecular, tissue, and cellular levels. This study focuses on the role of total antioxidant capacity (TAC) as a biomarker to be interpreted according to several clinical scenarios. After a brief description of various assay methods to elucidate terminology and physiopathological roles, we focus on the hormonal influence on TAC in blood plasma and other biological fluids, as different endocrine systems can modulate the antioxidant response. Furthermore, OS characterizes several endocrinopathies through different mechanisms: an inadequate antioxidant response to an increase in reducing equivalents (reductive distress) or a marked consumption of antioxidants (oxidative distress), which leads to low TAC values. An increased TAC could instead represent an adaptive mechanism, suggesting a situation of OS. Hence, the clinical context is fundamental for a correct interpretation of TAC. This review aims to provide the reader with a general overview of oxidative stress in several clinical examples of endocrine relevance, such as metabolic syndrome, non-thyroid illness syndrome, hypopituitarism, and infertility. Finally, the impact of dietary and surgical interventions on TAC in the model of metabolic syndrome is highlighted, along with personal experience. [ABSTRACT FROM AUTHOR]

Magnetotactic bacteria (MTB) are promising candidates for use as biomicrorobots in biomedical applications due to their motility, self-propulsion, and the ability to direct their navigation with an applied magnetic field. When in the body, the MTB may encounter non-Newtonian fluids such as blood plasma or mucus. However, their motility and the effectiveness of directed navigation in non-Newtonian fluids has yet to be studied on a single-cell level. In this work, we investigate motility of Magnetospirillum magneticum AMB-1 in three concentrations of polyacrylamide (PAM) solution, a mucus-mimicking fluid. The swimming speeds increase from 44.0 ± 13.6 µm/s in 0 mg/mL of PAM to 52.73 ± 15.6 µm/s in 1 mg/mL then decreases to 24.51 ± 11.7 µm/s in 2 mg/mL and 21.23 ± 10.5 µm/s in 3 mg/mL. This trend of a speed increase in low polymer concentrations followed by a decrease in speed as the concentration increases past a threshold concentration is consistent with other studies of motile, flagellated bacteria. Past this threshold concentration of PAM, there is a higher percentage of cells with an overall trajectory angle deviating from the angle of the magnetic field lines. There is also less linearity in the trajectories and an increase in reversals of swimming direction. Altogether, we show that MTB can be directed in polymer concentrations mimicking biological mucus, demonstrating the influence of the medium viscosity on the linearity of their trajectories which alters the effective path that could be predefined in Newtonian fluids when transport is achieved by magnetotaxis. [ABSTRACT FROM AUTHOR]

Abstract Background Previous research has indicated that extracellular vesicles (EVs) potentially play significant roles in multiple ageing phenotypes. This study uses a factorial experimental design to explore the interactions between circulating EVs and bone marrow-derived macrophages (BMDMs) isolated from young (7–12 weeks) and aged (70–90 weeks) mice. Results In this study, plasma EVs from young (Y_EV) and aged (O_EV) mice were isolated and compared based on abundance, size, and miRNA cargo. Compared to some previous studies, we found relatively few differences in EV miRNA cargo between Y_EVs and O_EVs. Young and old EVs were then used to stimulate naïve BMDMs isolated from young (Y_BMDM) and aged (O_BMDM) mice. A panel of five “M1” and six “M2” macrophage markers were used to assess the degree of polarisation. Our results revealed differences in the immunomodulatory effects of Y_EVs and O_EVs in Y_BMDMs and O_BMDMs. Y_EVs induced less pro-inflammatory gene expression, while O_EVs exhibited a more varied impact, promoting both pro- and anti-inflammatory markers. However, neither EV population induced a clearly defined ‘M1’ or ‘M2’ macrophage phenotype. We also report that EVs elicited responses that differed markedly from those induced by whole plasma. Plasma from old mice had strong pro-inflammatory effects on Y_BMDMs, increasing Il1b, Nlrp3 and Tnfa. However, O_EVs did not have these effects, supporting current evidence that EVs are a separate component of circulating factors during ageing. More research is needed to elucidate specific factors involved in inflammageing processes. Conclusions Our findings reveal age-related differences in EV cargo and function, with young EVs tending to suppress inflammatory markers more effectively than aged EVs. However, this is not straightforward, and EVs often promoted both M1 and M2 markers. These results suggest that EVs are a distinct component of circulating factors and hold potential for therapeutic strategies aimed at mitigating age-related inflammation and immune dysregulation.

Introduction. In glial tumors, lipid metabolism becomes abnormal. Analysis of lipid metabolism components can be an important characteristic of molecular and genetic profile of gliomas.Aim. To determine the correlation between plasma lipidome profile and immunohistochemical characteristics of glial tumors and to evaluate clinical significance of blood lipid spectrum analysis in preoperative assessment of molecular profile of gliomas.Materials and methods. Immunohistochemical measurement of O-6-methylguanine-DNA-methyl transferase (MGMT), Ki-67, p53, IDH1 tumor markers was performed using the corresponding antibody clones. Composition of plasma lipids was assessed using thin layer chromatography.Results. Even at the early stages of gliomagenesis, significant differences in cholesterol ethers, lysophosphatidylcholines, phosphatidylcholine (PC)/ lysophosphatidylcholine (LPC) ratio, neutral lipids (NL)/phospholipids (PL) in the blood were observed. Significant correlations between Ki-67, MGMT tumor markers and the above-mentioned lipidome parameters were found. The PC/LPC, NL/PL ratios in the blood of the patients from the groups with higher (above 10 %) and lower (below 10 %) Ki-67 mitotic indexes compared to healthy individuals were significantly lower. Therefore, the values of lipidome parameters allow to indirectly assess proliferative activity of gliomas which can be used for preoperative diagnosis of these tumors. No significant differences in the plasma PC/LPC and NL/PL ratios were found between the groups with MGMT promoter methylation and without it. No indirect predictor criteria for MGMT were found.Conclusion. It is impossible to determine decreased epigenetic activity of corresponding transcripts and preoperative prognosis for alkylating agent therapy based on the parameters of plasma lipid metabolism.

This study investigated within-subject variability in the circulating metabolome under controlled conditions, and whether divergent exercise training backgrounds were associated with alterations in the circulating metabolome assessed in resting samples. Thirty-seven men comprising of endurance athletes (END; body mass, 71.0 ± 6.8 kg; fat-free mass index, 16.9 ± 1.1 kg/m²), strength athletes (STR; 94.5 ± 8.8 kg; 23.0 ± 1.8 kg/m²), and recreationally active controls (CON; 77.6 ± 7.7 kg; 18.1 ± 1.0 kg/m²) provided blood samples after an overnight fast on two separate occasions controlled for time of day of sampling, recent dietary intake, time since last meal, and time since last exercise training session. A targeted profile of metabolites, performed using liquid chromatography and mass spectrometry on plasma samples, identified 166 individual metabolites and metabolite features, which were analyzed with intraclass correlation coefficients, a multilevel principal component analysis, and univariate t tests adjusted for multiple comparisons. The median intraclass correlation coefficient was .49, with 46 metabolites displaying good reliability and 31 metabolites displaying excellent reliability. No difference in the abundance of any individual metabolite was identified within groups when compared between visits, but a combined total of 44 metabolites were significantly different (false discovery rate <0.05) between groups (END vs. CON, 42 metabolites; STR vs. CON, 10 metabolites; and END vs. STR, five metabolites). Under similar measurement conditions, the reliability of resting plasma metabolite concentrations varies largely at the level of individual metabolites with ~48% of metabolites displaying good-to-excellent reliability. However, a history of exercise training was associated with alterations in the abundance of ~28% of metabolites in the targeted profile employed in this study. [ABSTRACT FROM AUTHOR]

Introduction: There are several types of surface treatments (coatings) aimed at improving the biocompatibility of cardiopulmonary bypass (CPB) circuit. Some coatings appear to require higher doses of heparin to maintain anticoagulation goals, and some of them might induce postoperative coagulopathy. In this study, we compared the amount of heparin required, postoperative bleeding, and inflammatory response according to three types of coatings. Method: We retrospectively included 300 consecutive adult patients who underwent cardiac surgery with CPB and received one of three coatings (Phisio®, Trillium®, and Xcoating™). Our primary objective was to compare, according to coating, the amount of heparin required to maintain an ACT > 400s during CPB. Our secondary objectives were to compare postoperative bleeding for 48 h and CRP rate. Results: Baseline characteristics were comparable between groups except for age and preoperative CRP. We did not find a significant difference between the 3 coatings regarding the amount of heparin reinjected. However, we found less postoperative bleeding with the Xcoating™ circuit compared to the Phisio® circuit (−149 mL [−289; −26.5]; p = 0.02) and a lower elevation of CRP with the Phisio® circuit (2.8 times higher than preoperative CRP) compared to Trillium® (4.9 times higher) and Xcoating™ (6.4 times higher); p < 10–3. Conclusion: The choice of coating did not influence the amount of heparin required during CPB; however, the post-CPB inflammatory syndrome may be impacted by this choice. [ABSTRACT FROM AUTHOR]

Dermatomyositis (DM) is a chronic multi-systemic inflammatory disorder of autoimmune origin, which has been associated with cardiovascular complications, including ventricular arrhythmias and sudden cardiac death. The Tp-e interval and Tp-e/QT ratio have been accepted as new markers for the assessment of myocardial repolarization and ventricular arrhythmogenesis. The aim of this study was to evaluate ventricular repolarization by using Tp-e interval and Tp-e/QT ratio in patients with DM, and to assess the relation with inflammation and autoimmunity. This study included 281 DM patients (180 females, 101 males; mean age 52.73 ± 15.80 years) and 281 control subjects (180 females, 101 males; mean age 53.38 ± 15.72 years). QTc, Tp-e interval and Tp-e/QT ratio were measured from the 12-lead ECG. The plasma level of blood routine test, C-reactive protein (CRP), erythrocyte sedimentation rate (ESR) was measured. These parameters were compared between groups. No statistically significant difference was found between two groups in terms of basic characteristics. In electrocardiographic parameters analysis, QTc, Tp-e interval and Tp-e/QT ratio were significantly increased in DM patients compared to the control group (441.44 ± 26.62 ms vs 422.72 ± 11.7 ms, 104.16 ± 24.34 ms vs 77.23 ± 16.25 ms and 0.27 ± 0.06 ms vs 0.20 ± 0.04 ms, all P value < 0.01). QTc, Tp-e interval and Tp-e/QT were positively correlated with NLR, CRP, and ESR (all P values < 0.01), and were increased in anti-Ro/SSA-52kD positive patients compared to those negative (452.33 ± 24.89 ms vs 438.55 ± 26.37 ms, 114.05 ± 22.68 ms vs 101.53 ± 24.13 ms, and 0.29 ± 0.06 ms vs 0.27 ± 0.05 ms, all P value < 0.01). Our study demonstrated that QTc, Tp-e interval, and Tp-e/QT ratio were increased in DM patients and were associated with inflammatory markers and anti-Ro/SSA-52kD positivity. [ABSTRACT FROM AUTHOR]

Deuterium oxide (D2O) appearance in blood is a marker of fluid bioavailability. However, whether biomarker robustness (e.g., relative fluid delivery speed) is consistent across analytical methods (e.g., cavity ring-down spectroscopy) remains unclear. Fourteen men ingested fluid (6 ml/kg body mass) containing 0.15 g/kg D2O followed by 45 min blood sampling. Plasma (D2O) was detected (n = 8) by the following: isotope-ratio mass spectrometry after vapor equilibration (IRMS-equilibrated water) or distillation (IRMS-plasma) and cavity ring-down spectroscopy. Two models calculated D2O halftime to peak (t1/2max): sigmoid curve fit versus asymmetric triangle (TRI). Background (D2O) differed (p <.001, η2 =.98) among IRMS-equilibrated water, IRMS-plasma, and cavity ring-down spectroscopy (152.2 ± 0.8, 147.2 ± 1.5, and 137.7 ± 2.2 ppm), but did not influence (p >.05) D2O appearance (Δppm), time to peak, or t1/2max. Stratifying participants based on mean t1/2max (12 min) into "slow" versus "fast" subgroups resulted in a 5.8 min difference (p <.001, η2 =.73). Significant t1/2max model (p =.01, η2 =.44) and Model × Speed Subgroup interaction (p =.005, η2 =.50) effects were observed. Bias between TRI and sigmoid curve fit increased with t1/2max speed: no difference (p =.75) for fast (9.0 min vs. 9.2 min, respectively) but greater t1/2max (p =.001) with TRI for the slow subgroup (16.1 min vs. 13.7 min). Fluid bioavailability markers are less influenced by which laboratory method is used to measure D2O as compared with the individual variability effects that influence models for calculating t1/2max. Thus, TRI model may not be appropriate for individuals with slow fluid delivery speeds. [ABSTRACT FROM AUTHOR]

Abstract The clinical diagnosis of biliary atresia (BA) poses challenges, particularly in distinguishing it from cholestasis (CS). Moreover, the prognosis for BA is unfavorable and there is a dearth of effective non-invasive diagnostic models for detection. Therefore, the aim of this study is to elucidate the metabolic disparities among children with BA, CS, and normal controls (NC) without any hepatic abnormalities through comprehensive metabolomics analysis. Additionally, our objective is to develop an advanced diagnostic model that enables identification of BA. The plasma samples from 90 children with BA, 48 children with CS, and 47 NC without any liver abnormalities children were subjected to metabolomics analysis, revealing significant differences in metabolite profiles among the 3 groups, particularly between BA and CS. A total of 238 differential metabolites were identified in the positive mode, while 89 differential metabolites were detected in the negative mode. Enrichment analysis revealed 10 distinct metabolic pathways that differed, such as lysine degradation, bile acid biosynthesis. A total of 18 biomarkers were identified through biomarker analysis, and in combination with the exploration of 3 additional biomarkers (LysoPC(18:2(9Z,12Z)), PC (22:5(7Z,10Z,13Z,16Z,19Z)/14:0), and Biliverdin-IX-α), a diagnostic model for BA was constructed using logistic regression analysis. The resulting ROC area under the curve was determined to be 0.968. This study presents an innovative and pioneering approach that utilizes metabolomics analysis to develop a diagnostic model for BA, thereby reducing the need for unnecessary invasive examinations and contributing to advancements in diagnosis and prognosis for patients with BA.

Introduction: The human plasma glycoproteome holds enormous potential to identify personalized biomarkers for diagnostics. Glycoproteomics has matured into a technology for plasma N-glycoproteome analysis but further evolution towards clinical applications depends on the clinical validity and understanding of protein- and site-specific glycosylation changes in disease. Objectives: Here, we exploited the uniqueness of a patient cohort of genetic defects in well-defined glycosylation pathways to assess the clinical applicability of plasma N-glycoproteomics. Methods: Comparative glycoproteomics was performed of blood plasma from 40 controls and 74 patients with 13 different genetic diseases that impact the protein N-glycosylation pathway. Baseline glycosylation in healthy individuals was compared to reference glycome and intact transferrin protein mass spectrometry data. Use of glycoproteomics data for biomarker discovery and sample stratification was evaluated by multivariate chemometrics and supervised machine learning. Clinical relevance of site-specific glycosylation changes were evaluated in the context of genetic defects that lead to distinct accumulation or loss of specific glycans. Integrated analysis of site-specific glycoproteome changes in disease was performed using chord diagrams and correlated with intact transferrin protein mass spectrometry data. Results: Glycoproteomics identified 191 unique glycoforms from 58 unique peptide sequences of 34 plasma glycoproteins that span over 3 magnitudes of abundance in plasma. Chemometrics identified high-specificity biomarker signatures for each of the individual genetic defects with better stratification performance than the current diagnostic standard method. Bioinformatic analyses revealed site-specific glycosylation differences that could be explained by underlying glycobiology and protein-intrinsic factors. Conclusion: Our work illustrates the strong potential of plasma glycoproteomics to significantly increase specificity of glycoprotein biomarkers with direct insights in site-specific glycosylation changes to better understand the glycobiological mechanisms underlying human disease.

Introduction. The number of peptide drugs being developed and registered has increased in recent years. Therefore, modern analytical approaches and methods are required to determine these substances in biological matrices during pharmacokinetic studies. Peptides are structurally intermediate between small molecules and biopolymers, making it difficult to develop methods for determining them using High Performance Liquid Chromatography with Tandem Mass Spectrometry (HPLC-MS/MS). Peptide derivatization can help achieve optimal chromatographic separation and increase method sensitivity.Aim. To develop and validate a method for the determination of the tetradecapeptide (TDP) threonyl-glutamyl-lysyl-lysyl-arginyl-arginyl-glutamayl-threonyl-valyl-glutamyl-arginyl-glutamyl-lysyl-glutamate in human plasma by HPLC-MS/MS.Materials and methods. The determination of TDP in human plasma was performed by HPLC-MS/MS. Sample preparation included a combination of blood plasma protein precipitation with propionic acid solution in methanol, liquid-liquid extraction with chloroform, and peptide derivatization with propionic anhydride. Internal standard (IS) was threonyl-glutamyl-lysyl-lysyl-arginyl-arginyl-glutamayl-threonyl-leucyl-glutamyl-arginyl-glutamyl-lysyl-glutamate. Chromatographic separation was performed in gradient mode, eluent A was 0.1 % formic acid solution in water, eluent B was 0.1 % formic acid in acetonitrile. Column: Waters XBridge C18, 4.6 × 50 mm, 5 µm. Ionization source was electrospray in positive mode. Multiple reaction monitoring (MRM) transitions for 4-substituted TDP propionate were: 681.30 → 73.95 m/z, 681.30 → 84.00 m/z, 681.30 → 101.90 m/z, 681.30 → 140.10 m/z, and for 4-substituted IS propionate: 686.00 → 74.10 m/z, 686.00 → 84.05 m/z, 686.00 → 102.00 m/z, 686.00 → 140.00 m/z.Results and discussion. Validation of the developed method was carried out in accordance with the requirements of Eurasian Economic Union and the following parameters were determined: selectivity, matrix effect, calibration curve, accuracy and precision, recovery, lower limit of quantification, sample carryover, stability.Conclusion. The method for the determination of TDP in human blood plasma by HPLC-MS/MS was developed and validated. The analytical range was 5.00–1000.00 ng/mL, allowing the method to be used to study TDP pharmacokinetics.

Abstract Periampullary cancers, including pancreatic ductal adenocarcinoma, ampullary-, cholangio-, and duodenal carcinoma, are frequently diagnosed in an advanced stage and are associated with poor overall survival. They are difficult to differentiate from each other and challenging to distinguish from benign periampullary disease preoperatively. To improve the preoperative diagnostics of periampullary neoplasms, clinical or biological markers are warranted. In this study, 28 blood plasma amino acids and derivatives from preoperative patients with benign (N = 45) and malignant (N = 72) periampullary disease were analyzed by LC-MS/MS. Principal component analysis and consensus clustering both separated the patients with cancer and the patients with benign disease. Glutamic acid had significantly higher plasma expression and 15 other metabolites significantly lower plasma expression in patients with malignant disease compared with patients having benign disease. Phenylalanine was the only metabolite associated with improved overall survival (HR = 0.50, CI 0.30–0.83, P

Background: Inherited antithrombin, protein C, and protein S deficiency increase the risk of venous thromboembolism. The presence of defects can be identified by clinical laboratory assays. In most Chinese clinical laboratories, the screening tests for antithrombin, protein C, and protein S deficiency are their activity assays. Ensuring appropriate pre-analytical storage conditions for activity tests is essential. This study aimed to assess the effects of storage conditions on antithrombin, protein C, and protein S activity in frozen plasma. Methods: We collected the remaining plasma of 29 patients. The baseline of antithrombin, protein C, and protein S activity values were tested within 4 h. Then, each sample was sub-packaged into 4 EP tubes, and was stored at -20 °C for 3 days, -20 °C for 7 days, -80 °C for 3 days, and − 80 °C for 7 days, respectively. After thawing, samples were tested by two systems. Results: The percentage deviation of antithrombin and protein C activity assay was<10% compared with the initial values. Protein S activity showed a significant reduction in frozen plasma, with a deviation > 10%. Some samples, initially within the normal range, were classified as abnormal after freezing storage. Conclusions: Our study indicated that antithrombin and protein C remain stable when stored at -20 °C or -80 °C in a week. We argued that Protein S activity is not stable in frozen plasma. The use of frozen-thawed plasma for PS activity assay may result in overdiagnosis of protein S deficiency. [ABSTRACT FROM AUTHOR]

Background: Epstein-Barr virus DNA (EBV-DNA) is closely related to the pathogenesis and prognosis of EBV-associated hemophagocytic lymphohistiocytosis (EBV-HLH). The quantitative measurement of blood EBV-DNA is widely used in EBV-HLH, but there remains a lack of evidence to guide clinicians. Methods: A retrospective analysis was conducted on clinical manifestations, laboratory tests, 310 blood EBV-DNA loads, and prognosis of 51 pediatric patients diagnosed with EBV-HLH. Receiver operating characteristic (ROC) curves were utilized to determine the optimal cutoff values of EBV-DNA for predicting mortality and evaluating the active status of EBV-HLH. Results: EBV-positive- lymphoma-HLH had higher initial plasma EBV-DNA load(1.10 × 106copies/ml) compared to the EBV-HLH group (1.98 × 104 copies/ml) (P = 0.006), and experienced recurrently elevated plasma EBV-DNA levels during treatment. The optimal cut-off value of initial plasma EBV-DNA load in predicting mortality was 2.68 × 105 copies/ml, with a sensitivity of 88.57% and a specificity of 56.25%. For determining the active status of HLH, the optimal cutoff value of PBMC EBV-DNA load during treatment was 2.95 × 105 copies/ml, with a sensitivity of 69.14% and a specificity of 64.71%. The cut-off value of plasma EBV-DNA for determining active status was 1.32 × 103 copies/ml, with a sensitivity of 84.34%, and a specificity of 87.67%. Patients with higher PBMC and plasma EBV-DNA at initial and those with repeated elevated plasma EBV-DNA during treatment had worse prognoses (P < 0.05). Conclusion: Dynamic monitoring of EBV-DNA is a valuable tool for assessing disease status and predicting the prognosis of EBV-HLH, with plasma EBV-DNA being more effective than PBMC EBV-DNA. Patients with high levels of PBMC and plasma EBV-DNA at initial and those with repeated elevated plasma EBV-DNA during treatment had worse prognoses. [ABSTRACT FROM AUTHOR]

Robust circadian rhythms are essential for optimal health. The central circadian clock controls temperature rhythms, which are known to organize the timing of peripheral circadian rhythms in rodents. In humans, however, it is unknown whether temperature rhythms relate to the organization of circadian rhythms throughout the body. We assessed core body temperature amplitude and the rhythmicity of 929 blood plasma metabolites across a 40-h constant routine protocol, controlling for behavioral and environmental factors that mask endogenous temperature rhythms, in 23 healthy individuals (mean [± SD] age = 25.4 ± 5.7 years, 5 women). Valid core body temperature data were available in 17/23 (mean [± SD] age = 25.6 ± 6.3 years, 1 woman). Individuals with higher core body temperature amplitude had a greater number of metabolites exhibiting circadian rhythms (R2 = 0.37, p =.009). Higher core body temperature amplitude was also associated with less variability in the free-fitted periods of metabolite rhythms within an individual (R2 = 0.47, p =.002). These findings indicate that a more robust central circadian clock is associated with greater organization of circadian metabolite rhythms in humans. Metabolite rhythms may therefore provide a window into the strength of the central circadian clock. [ABSTRACT FROM AUTHOR]

Background: Data show that due to endothelial damage and thrombogenic effects, severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection may accelerate the development of atherosclerosis and increase the risk of cardiovascular diseases (CVDs). The impaired metabolism of aminothiols increases oxidative stress, as these molecules are involved in antioxidant defense as well as in thiol redox control. In this study, total levels of selected aminothiols (i.e., cysteine (Cys), homocysteine (HCy), and glutathione) in convalescents after coronavirus disease of 2019 (COVID-19) were evaluated. The analyses were made according to the sex of the patients, time from COVID-19 onset, and COVID-19 severity. Methods: The study group consisted of 212 patients after COVID-19. Levels of total aminothiols were assessed in the blood plasma using high-performance liquid chromatography (HPLC). Results: The mean Cys concentrations were higher in men than in women (229.92 µmol/L ± 51.54 vs. 210.35 µmol/L ± 41.90, respectively; p = 0.003). Differences in Cys levels were also noticed in the total study group between patients distinguished due to time from disease onset (226.82 µmol/L ± 40.57 in <12 weeks, 232.23 µmol/L ± 47.99 in patients 12–24 weeks, and 208.08 µmol/L ± 48.43 in patients >24 weeks; p = 0.005). In addition, over 11% of total patients 12–24 weeks from disease onset had Cys levels above 300 µmol/L compared to almost 4% of patients <12 weeks and 2% of patients >24 weeks (p = 0.046). In sex-adjusted subgroups, significant differences due to time from COVID-19 were found in Cys levels in women (p = 0.004) and in glutathione levels in men (p = 0.024). None of the aminothiol levels differed between the subgroups based on the severity of COVID-19. Conclusions: Men had overall higher Cys levels than women. Cys levels were lower >24 weeks after COVID-19 onset than in the earlier period after disease onset. A partial elevation in Cys levels 12–24 weeks after the disease onset may contribute to the increase in CVD risk in the post-COVID-19 period. [ABSTRACT FROM AUTHOR]

Cytokine storm is usually described as one of the main reasons behind COVID-associated mortality. Cytokines are essential protein molecules engaged in immune responses; they play a critical role in protection against infections. However, they also contribute to inflammatory reactions and tissue damage, becoming a double-edged sword in the context of COVID-19. Recent studies have suggested various cytokines and chemokines that play a crucial role in the immune response to SARS-CoV-2 infection. One such cytokine is interleukin 27 (IL-27), which has been found to be elevated in the blood plasma of patients with COVID-19. Within this study, we will explore the role of IL-27 in immune responses and analyze both the existing literature and our own prior research findings on this cytokine in the context of COVID-19. It affects a wide variety of immune cells. Regardless of the pathological process it is involved in, IL-27 is critical for upholding the necessary balance between tissue damage and cytotoxicity against infectious agents and/or tumors. In COVID-19, it is involved in multiple processes, including antiviral cytotoxicity via CD8+ cells, IgG subclass switching, and even the activation of Tregs. [ABSTRACT FROM AUTHOR]

Temporal lobe epilepsy has various origins, involving or not involving structural changes in brain tissue. The mechanisms of epileptogenesis are associated with cell regulation and signaling disruptions expressed in varied levels of proteins. The blood plasma proteomic profiling of temporal lobe epilepsy patients (including magnetic resonance imaging (MRI)-positive and MRI-negative ones) and healthy volunteers using mass spectrometry and label-free quantification revealed a list of differently expressed proteins. Several apolipoproteins (APOA1, APOD, and APOA4), serpin protease inhibitors (SERPINA3, SERPINF1, etc.), complement components (C9, C8, and C1R), and a total of 42 proteins were found to be significantly upregulated in the temporal lobe epilepsy group. A classification analysis of these proteins according to their biological functions, as well as a review of the published sources, disclosed the predominant involvement of the processes mostly affected during epilepsy such as neuroinflammation, intracellular signaling, lipid metabolism, and oxidative stress. The presence of several proteins related to the corresponding compensatory mechanisms has been noted. After further validation, the newly identified temporal lobe epilepsy biomarker candidates may be used as epilepsy diagnostic tools, in addition to other less specific methods such as electroencephalography or MRI. [ABSTRACT FROM AUTHOR]

Rapid and safe hemostasis is crucial for the survival of bleeding patients in prehospital care. It is urgent to develop high performance hemostatic material to control the massive hemorrhage in the military field and accidental trauma. In this work, an efficient protein hemostat of thrombin was immobilized onto commercial gauze, which was mediated by self-polymerization and anchoring of tannic acid (TA). Through TA treatment, the efficient immobilization of thrombin was achieved, preserving both the biological activity of thrombin and the physical properties of the dressing, including absorbency, breathability, and mechanical performance. Moreover, in the presence of TA coating and thrombin, Gau@TA/Thr could obviously shortened clotting time and enriched blood components such as plasma proteins, platelets, and red blood cells, thereby exhibiting an enhanced in vitro coagulation effect. In SD rat liver volume defect and artery transection hemorrhage models, Gau@TA/Thr still had outstanding hemostatic performance. Besides, the Gau@TA/Thr gauze had inherent antibacterial property and demonstrated excellent biocompatibility. All results suggested that Gau@TA/Thr would be a potential candidate for treating uncontrollable hemorrhage in prehospital care. [ABSTRACT FROM AUTHOR]

Human serum albumin (HSA), a polypeptide featuring 17 disulfide bonds, acts as a crucial transport protein in human blood plasma. Its extended circulation half-life, mediated by FcRn (neonatal Fc receptor)-facilitated recycling, positions HSA as an excellent carrier for long-acting drug delivery. However, the conventional method of obtaining HSA from human blood faces limitations due to availability and potential contamination risks, such as blood-borne diseases. This study introduced SHuffle, an oxidative Escherichia coli (E. coli) expression system, for the production of recombinant HSA (rHSA) that spontaneously self-folds into its native conformation. This system ensures precise disulfide bond formation and correct folding of cysteine-rich rHSA, eliminating the need for chaperone co-expression or domain fusion of a folding enhancer. The purified rHSA underwent thorough physicochemical characterization, including mass spectrometry, circular dichroism spectroscopy, intrinsic fluorescence spectroscopy, esterase-like activity assay, and size exclusion chromatography, to assess critical quality attributes. Importantly, rHSA maintained native binding affinity to FcRn and the albumin-binding domain. Collectively, our analyses demonstrated a high comparability between rHSA and plasma-derived HSA. The expression of rHSA in E. coli with an oxidizing cytosol provides a secure and cost-effective approach, enhancing the potential of rHSA for diverse medical applications. • E. coli with an oxidative cytoplasm produced recombinant human serum albumin (rHSA) without solubility tags or chaperones. • rHSA exhibited good comparability to plasma-derived HSA in structure and function. • rHSA retained the native ability of HSA to bind the albumin-binding domain (ABD) and human FcRn. [ABSTRACT FROM AUTHOR]

Physical properties of blood plasma, such as viscosity, serve as crucial indicators of disease. The inherent capillary effect of paper microchannels, coupled with minimal sample requirement, stimulated the advancement of paper-based viscometers. This study presents a precise, non-contact optoelectronic system using a microfluidic platform for the measurement of blood plasma viscosity. Microchannels were defined onto the filter paper using an available and inexpensive wax crayon, without the need for conventional wax printing equipment. The time required for the 5 μL sample to pass a specific distance was measured using two pairs of infrared sensors. Subsequently, this data was sent to the microcontroller, which automatically calculated the viscosity. Throughout the measurements, sample temperature was maintained at a constant 37 °C through an integrated heater with automated control. The microfluidic platform successfully processed real samples, yielding viscosity measurements in under three minutes. Evaluation with fetal bovine serum, spiked with varying protein concentrations in both native and denatured states, demonstrated a precision exceeding 96% compared to conventional Ostwald viscometer readings. For human subjects exhibiting pathologies affecting serum and plasma viscosity compared to physiological norms, strong correlations were observed between resultant values and clinical diagnoses. The proposed device aims to replace expensive and complex optical equipment, offering a safer alternative for measuring plasma viscosity. Unlike similar devices, it eliminates the risk of component deformation due to chemical contact or unsafe irradiation. [ABSTRACT FROM AUTHOR]

AbstractReproduction is a fundamental biological process for producing offspring and ensuring the continuation of the species. With nearly 35,000 fish species worldwide exhibiting a very diverse range of reproductive systems and strategies, the study of fish reproduction is a significant challenge. Research into various fish reproductive models is driven by their use in human biomedical applications, improving the diversification and sustainability of aquaculture species, and developing strategies for biodiversity conservation. These models must be approached from an animal ethics perspective, considering the welfare of the species under study and applying the 3Rs principle (Replacement, Reduction and Refinement). This principle focusses on replacing animals with alternative methods wherever possible, reducing the number of animals used and refining procedures to minimize suffering. While destructive sampling of wild aquatic species should be minimized, the need remains to provide information on reproduction as an important parameter for delineating management and conservation programs. One of the practices that helps reduce and replace the use of animal testing is the scientific study of individuals obtained through bycatch. This is the case for some chondrichthyans, and endangered fish species included in conservation programs. Regarding fish species that are kept under human care (commonly called captivity), studies on reproduction are essential to improve well-being and increase production of farmed species. Research on fish reproduction requires expanding basic knowledge of broodstock management tools, such as gamete quality assessment, reproductive behavior studies, the use of hormone therapies to stimulate reproduction, and the development of non-lethal or less invasive methods using body fluids (e.g., mucus, seminal plasma, blood plasma, or urine) to study reproductive status, as well as novel markers from state-of-the-art omics research. This review describes a multidisciplinary approach that includes the aforementioned reproductive management tools, indicators of welfare, and next-generation sequencing techniques using samples collected by minimally invasive methods. [ABSTRACT FROM AUTHOR]

Distant hybridization is a well-established technique in genetic breeding for cultivation of superior traits. Previous studies have shown that, hybrids derived from the cross between Schizothorax wangchiachii (♀) and Percocypris pingi (♂) are significantly different from their parents in terms of growth performance, feeding behavior and disease resistance. Both cellular and plasma components of blood are integral to immune system, and erythrocytes serve as the sole cells for transporting oxygen which makes energy available for the growth, development, immune responses and all other physiological activities. In order to understand ecological adaptive strategies and also the underlying physiological and biochemical mechanisms of fish with different characteristics, the present study used offsprings and hybrids produced by artificial reproduction of the same batch of domesticated S. wangchiachii, P. pingi broodstock as the experimental fish, and analyses of biochemical indexes, differential leucocytes counts and histological examination of liver sections were performed. Results showed that the proportions of lymphocytes and thrombocytes in P. pingi were significantly higher than those in S. wangchiachii (P<0.05), while the proportions of granulocytes and monocytes in S. wangchiachii were significantly higher than those in P. pingi (P<0.05). In contrast, all blood cell values of hybrids fell between those of its parent species, with the exception of the percentage of thrombocytes, which was the highest of the three. The erythrocyte area, perimeter and short diameter of hybrid fish were significantly higher than those of S. wangchiachii and P. pingi (P<0.0001), while P. pingi exhibited the smallest erythrocyte values. Hepatocyte area, perimeter and short diameter were also significantly higher in hybrids than in S. wangchiachii and P. pingi, and P. pingi had the smallest hepatocyte area, perimeter, long diameter and short diameter. However, size and perimeter of hepatic nucleus of hybrids were the smallest. The number of Kupffer cells per unit area of liver was significantly greater in S. wangchiachii than in hybrids and Percocypris pingi (P<0.05). In terms of biochemical markers, plasma C-reactive protein level was nearly undetectable in S. wangchiachii, but it could reach 10 mg/L in both P. pingi and hybrids. Besides, plasma total bile acids was significantly higher in S. wangchiachii than in P. pingi (P<0.01), and hybrids fell in intermediate level. These results suggest that there may be differences in the immune functions among S. wangchiachii, P. pingi, and their hybrids, as well as in their liver metabolism and protein synthesis capabilities. These findings here might provide some crucial clues in understanding the biodiversity and fish adaptation, further promote the development of wild fish cultivation. [ABSTRACT FROM AUTHOR]

Background: The function of polymorphonuclear neutrophils (PMNs) decreases with age, which results in infectious and inflammatory complications in older individuals. The underlying causes are not fully understood. ATP release and autocrine stimulation of purinergic receptors help PMNs combat microbial invaders. Excessive extracellular ATP interferes with these mechanisms and promotes inflammatory PMN responses. Here, we studied whether dysregulated purinergic signaling in PMNs contributes to their dysfunction in older individuals. Results: Bacterial infection of C57BL/6 mice resulted in exaggerated PMN activation that was significantly greater in old mice (64 weeks) than in young animals (10 weeks). In contrast to young animals, old mice were unable to prevent the systemic spread of bacteria, resulting in lethal sepsis and significantly greater mortality in old mice than in their younger counterparts. We found that the ATP levels in the plasma of mice increased with age and that, along with the extracellular accumulation of ATP, the PMNs of old mice became increasingly primed. Stimulation of the formyl peptide receptors of those primed PMNs triggered inflammatory responses that were significantly more pronounced in old mice than in young animals. However, bacterial phagocytosis and killing by PMNs of old mice were significantly lower than that of young mice. These age-dependent PMN dysfunctions correlated with a decrease in the enzymatic activity of plasma ATPases that convert extracellular ATP to adenosine. ATPases depend on divalent metal ions, including Ca2+, Mg2+, and Zn2+, and we found that depletion of these ions blocked the hydrolysis of ATP and the formation of adenosine in human blood, resulting in ATP accumulation and dysregulation of PMN functions equivalent to those observed in response to aging. Conclusions: Our findings suggest that impaired hydrolysis of plasma ATP dysregulates PMN function in older individuals. We conclude that strategies aimed at restoring plasma ATPase activity may offer novel therapeutic opportunities to reduce immune dysfunction, inflammation, and infectious complications in older patients. [ABSTRACT FROM AUTHOR]

Zinc plays a central role in many metabolic processes in the organism and is therefore one of the most important trace elements in horse nutrition. Therefore, the adequate supply and supplementation of zinc is essential for maintaining the health and performance of horses. In this study, the trace element concentration of zinc in different blood sample media will be compared and analyzed with those in faecal samples. Furthermore, reference values for the zinc concentration in fecal samples are to be established in order to present a valid, minimally invasive, diagnostic alternative to the evaluation of blood serum. It should also be critically examined whether the coprological examination is preferable to the serological examination. The evaluation of the trace element concentration of zinc was carried out using a standardized, analytical procedure. Blood (serum, plasma and EDTA blood) and faecal samples were taken from 10 test subjects from 13 federal states, analyzed and compared. The test subjects were horses of all breeds, aged between 3 and 35 years, which were classified as clinically healthy.[1] Furthermore, the horses were used both recreationally and for sport. The horses came from different farms and were kept in conventional stables, open stables and year-round grazing. There was no uniform feeding of concentrates and supplementary feed. The feeding of copper-containing mineral feed was detected in some horses, but was not considered separately in this study. The zinc concentration was analyzed using ICP)OES (optical emission spectrometry with inductively coupled plasma). The faecal samples were prepared and dried beforehand. There was no biologically relevant correlation between the zinc concentration in fecal and serum samples. In the blood samples, however, a strong correlation could be demonstrated between serum and plasma samples, but not between serum and EDTA blood samples. The reference values determined for the zinc concentration in fecal samples are between 29 and 413mg/kg. Reference values for the zinc concentration in fecal samples were determined. Staufenbiel et al. already postulated the first methodological framework for this. Among other things, the coprological examination is the less invasive diagnostic tool. Furthermore, unlike the serological examination, the fecal sample examination provides information about the nutritive supply basis of zinc, which means that a more precise statement can be made about the zinc concentration in the mechanism. Due to its practicability, fecal sample analysis is preferable to serological evaluation. [ABSTRACT FROM AUTHOR]

Objectives: This study aimed to investigate the potential protective impact of anthocyanin against tissue damage and oxidative stress provoked by varicocele within the testes of adult rats. Materials and Methods: A total of 32 male rats were divided into four groups as follows: a control group undergoing a sham procedure, a varicocele group without intervention (V), a varicocele group treated with anthocyanin (VA), and a group receiving anthocyanin treatment alone. Following a 56-day treatment period, the indicators of oxidative stress were gauged in the blood plasma, while histological modifications were evaluated utilizing the hematoxylin and eosin staining technique. Results: In the varicocele group treated with anthocyanin, we observed noteworthy enhancements in Johnsen score, epithelium thickness, and seminiferous tubule diameter compared to the untreated varicocele group. Treatment groups exhibited substantial elevations in testosterone levels and antioxidant enzyme levels. Furthermore, there was a reduction in the levels of malondialdehyde (MDA), an established marker of oxidative damage, and a decline in histological damage in the treatment groups. Conclusions: The outcomes underscore the potential safeguarding influence of anthocyanin against testicular damage stemming from varicocele induction, suggesting its beneficial role in countering oxidative stress and tissue impairment. [ABSTRACT FROM AUTHOR]

Objective: It was previously shown that the levels of several biomarkers increase due to acromegaly disease-related inflammation, and some markers are parallel to the disease's activity. The current study compared the possible relationship of growth differentiation factor-15 (GDF-15) with acromegaly disease activity in healthy subjects. Methods: This case-control study was conducted in a single center. It included 40 acromegaly patients (25 active/15 controlled) (47.7 ± 9.4 years, 20 female/20 male) and 24 healthy individuals (49.9 ± 10.1 years, 13 female/11 male) with age-sex-body mass index similar to the patient group. Demographic data, metabolic and hormonal parameters, and GDF-15 levels of the study population were studied. Results: The median GDF-15 levels were significantly higher in patients with acromegaly compared to healthy subjects (HS) (280.4 (Q1-Q3: 197.0-553.2) vs. 213.3 (Q1-Q3: 179.9-297.2) ng/L, P = .01). Serum GDF-15 levels of active and controlled acromegaly patients were comparable (P = .39). Interestingly, compared to HS, GDF-15 levels were significantly higher in controlled disease (P = .013), whereas GDF-15 levels tended to be higher in active disease but did not reach statistical significance (P = .06). Growth differentiation factor-15 levels were positively correlated with fasting plasma glucose (r = 0.304, P = .01) and HbA1c (r = 0.292, P = .02). When evaluated across the entire cohort, GDF-15 levels were found to be higher in diabetic patients compared to non-diabetic individuals (P = .04). Conclusion: Plasma GDF-15 levels were increased in the patients with acromegaly compared to healthy subjects. This increment may be due to accompanying diseases such as diabetes rather than a disease-specific effect. [ABSTRACT FROM AUTHOR]

Objectives: Infertility is inability to conceive after 12 months of regular unprotected sex. MiRNA expression changes can serve as potential biomarkers for infertility in males due to impaired spermatogenesis. This research was conducted to measure the expression level of miR‐211 in plasma samples as a factor identifying infertility in comparison with the control group. Methods: In this study, blood plasma were taken from the infertile men (n = 103) nonobstructive azoospermia (NOA) or severe oligozoospermia (SO) and the control group (n = 121). The expression of circulating miR‐211 in plasma was assessed by qRT‐PCR. A relative quantification strategy was adopted using the 2−ΔΔCT method to calculate the target miR‐211 expression level in both study groups. Results: Plasma miR‐211 levels were significantly lower in infertile men compared to the control group (0.544 ± 0.028 and 1.203 ± 0.035, respectively, p < 0.001). Pearson's correlation analysis showed that miR‐211 expression level has a positive and significant correlation with sperm parameters, including sperm concentration, sperm total motility, progressive motility, and normal morphology (p < 0.001). Conclusions: Decreased expression of miR‐211 in blood plasma seems to be associated with male infertility. This experiment showed that miR‐211 can be considered as a biomarker for evaluation, diagnosis, and confirmation of the results of semen analysis in male infertility. [ABSTRACT FROM AUTHOR]

Peripheral cytokines may influence psychoemotional behavior, but the role of interleukin-1beta (IL-1beta) in altering anxiety and motor activity in response to inflammatory activation remains unclear. To clarify this issue, correlations between behavioral parameters in the elevated plus maze (EPM) test and plasma levels of IL-1beta after administration of the proinflammatory stimulus lipopolysaccharide (LPS) in different modes were analyzed in adult male rats. LPS in doses of 0.5 or 5 mg/kg, as well as physiological solution (control), were administered to rats intraperitoneally. The most pronounced behavioral effect 24 hours after a single injection was an endotoxin dose-dependent inhibition of the animals' motor activity. Twenty four hours after the 5 mg/kg, an increased anxiety behavior was also observed. The behavioral changes caused by the high dose of endotoxin were completely normalized after a week. The behavior of the animals one day after the end of repeated injections of LPS at a lower dose for a week (0.5 mg/kg; once every two days) also did not differ from the control. The inhibition of motor activity after LPS could be due to an increase in the level of IL-1beta in the blood plasma, as indicated by the identified significant negative correlations between IL-1beta and the corresponding behavioral parameters. No significant correlation was found between the peripheral level of IL-1beta and such a classic indicator of anxiety as the percentage of entries into the open arms of the maze. In general, the obtained results allow us to conclude that IL-1beta is an undoubted participant in the mechanism of the transient inhibitory effect of LPS on motor activity. [ABSTRACT FROM AUTHOR]

Understanding pharmacokinetics (PK) in children is a prerequisite to determine optimal pediatric dosing. As plasma sampling in children is challenging, alternative PK sampling strategies are needed. In this case study we evaluated the suitability of saliva as alternative PK matrix to simplify studies in infants, investigating metamizole, an analgesic used off‐label in infants. Six plasma and 6 saliva PK sample collections were scheduled after a single intravenous dose of 10 mg/kg metamizole. Plasma/saliva pharmacometric (PMX) modeling of the active metabolites 4‐methylaminoantipyrine (4‐MAA) and 4‐aminoantipyrine (4‐AA) was performed. Various reduced plasma sampling scenarios were evaluated by PMX simulations. Saliva and plasma samples from 25 children were included (age range, 5–70 months; weight range, 8.7–24.8 kg). Distribution of metamizole metabolites between plasma and saliva was without delay. Estimated mean (individual range) saliva/plasma fractions of 4‐MAA and 4‐AA were 0.32 (0.05–0.57) and 0.57 (0.25–0.70), respectively. Residual variability of 4‐MAA (4‐AA) in saliva was 47% (28%) versus 17% (11%) in plasma. A simplified sampling scenario with up to 6 saliva samples combined with 1 plasma sample was associated with similar PK parameter estimates as the full plasma sampling scenario. This case study with metamizole shows increased PK variability in saliva compared to plasma, compromising its suitability as single matrix for PK studies in infants. Nonetheless, rich saliva sampling can reduce the number of plasma samples required for PK characterization, thereby facilitating the conduct of PK studies to optimize dosing in pediatric patients. [ABSTRACT FROM AUTHOR]

Highlights: What are the main findings? The validated method was robust, reliable, and sensitive for quantifying ABZ, ABZ-OX, ABZ-ON, and PR simultaneously from the plasma and dried blood spots. The method is well suited for therapeutic drug monitoring due to its high sensitivity, minimal sample volume requirement, and the ability for remote collection. What is the implication of the main finding? Dried blood spot (DBS) analysis represents an alternative sampling technique over conventional blood-sampling methods, particularly for sample collection, enabling a non-clinical setup without requiring special equipment or trained healthcare personnel. Additionally, it minimizes the solvent volume and analysis time and reduces transportation and storage needs. Millions of individuals throughout the world suffer from lymphatic filariasis (LF), which is a morbid disease caused by Wuchereria bancrofti, Brugia malayi, and Brugia timori. These infections belong to tissue-invading nematodes and are one of the major neglected tropical diseases that often result in permanent and enduring disability among individuals in endemic regions. Due to combination therapy, LF eradication has drastically decreased infections globally. The development of blood micro-sampling techniques allowing precise quantitation of drugs in blood would facilitate pharmacokinetic (PK) studies in remote populations. Therefore, an LC-MS/MS bioanalytical method was utilized to analyze albendazole (ABZ), albendazole sulfone (ABZ-ON), albendazole sulfoxide (ABZ-OX), and probenecid (PR) in plasma and dried blood spots. Solid-phase extraction was utilized to extract the analyte from both plasma and blood-spiked DBS. Analytes of interest were eluted with a gradient mobile system using 0.05% formic acid in water (A) and 0.05% formic acid in methanol (B) and separated using a reversed-phase Acquity ®BEH C18 UPLC column (100 × 2.1 mm, 1.7 µm). Precision and accuracy at each QC level were within the acceptable limit, i.e., ±15% for all analytes in both the matrices. Tests for stability under laboratory and storage conditions indicated that no notable changes were observed for plasma and DBS. The LC-MS/MS method demonstrated its capability to consistently identify all target analytes (ABZ, ABZ-ON, ABZ-OX, and PR) at low concentrations, even at the small specimen volumes obtained from DBS cards. This confirms the efficacy and durability of DBS cards as a micro-sampling technique. Moreover, it enhances collection efforts for therapeutic drug monitoring in remote locations for patients infected with lymphatic filariasis. [ABSTRACT FROM AUTHOR]

We present a case of a man immunocompromised due to myelodysplastic syndrome with Candida krusei fungemia who had a rising cell-free DNA (cfDNA) giant magnetoresistance (GMR) signal when tested daily using plasma blood samples. With the rise in GMR signal paralleling the development of skin lesions in this patient, we conclude that cfDNA can be used to indicate uncontrolled infection and thus help monitor response to therapy. This index patient provides evidence that an invasive fungal infection requires both direct antifungal therapy and an intact immune system to control the infection. This biosensing platform has been simplified to potentially serve as a point-of-care test, setting it apart by overcoming the three common barriers of cfDNA testing: complexity, cost, and time. [ABSTRACT FROM AUTHOR]

Background: Equine parvovirus hepatitis (EqPV‐H) can cause Theiler's disease and subclinical hepatitis in horses. Objectives: Assess the frequency of subclinical EqPV‐H infection in hospitalized horses and to study viral transmission by investigating potential shedding routes. Animals: One hundred sixteen equids, that presented to the University Equine Hospital of the University of Veterinary Medicine Vienna between February 2021 and March 2022, for causes other than hepatopathy. Methods: In this cross‐sectional study, samples (serum, feces, nasal, and buccal swabs) of hospitalized horses were collected. Sera were screened for the presence of anti‐EqPV‐H antibodies by a luciferase immunoprecipitation system assay. Quantitative PCR was used for the detection of EqPV‐H DNA in the samples and a nested PCR was used for further validation. Results: Seroprevalence was 10.3% (12/116) and viremia occurred in 12.9% (15/116) of the serologically positive horses. The detected viral load in serum varied from non‐quantifiable amount to 1.3 × 106 genome equivalents per milliliter of serum. A low viral load of EqPV‐H DNA was detected in 2 nasal swabs and 1 fecal sample. Conclusion and Clinical Importance: EqPV‐H DNA was detected in nasal secretions and feces of viremic horses, which could pose a risk to naive hospitalized horses. It is advisable to screen hospitalized horses that are potential donors of blood or plasma to reduce the risk of iatrogenic EqPV‐H transmission. [ABSTRACT FROM AUTHOR]

Little is known about how plasma and whole blood taurine and plasma carnitine correlate to concentrations in skeletal and cardiac muscle and the effects of diet in dogs. The purpose of this study was to evaluate the correlation among plasma, skeletal and cardiac muscle carnitine and taurine and whole blood taurine and determine the effect of diet. The study protocol was approved by the Pet Food Solutions Institutional Animal Care and Use Committee. Thirty‐three mixed‐breed hounds and 32 beagles were evaluated at Day 0 then removed from their baseline diet and randomized to a test diet: high animal protein, grain‐inclusive (HA‐GI), low animal protein, grain‐free (LA‐GF), low animal protein, grain‐inclusive (LA‐GI), or high animal protein, grain‐free (HA‐GF). Blood was drawn every 30 days and endomyocardial (mixed breeds only) and skeletal muscle biopsies were collected at Days 0 and 180. The correlations between plasma and whole blood taurine, or plasma carnitine and skeletal and cardiac muscle concentrations were weak (p < 0.01–0.05). Mixed‐breed hounds had increased (p = 0.029) whole blood taurine compared to beagles. Plasma taurine was lower with diet HA‐GF, (p = 0.009) however, all diets had increased taurine from Day 0 and were, on average within the laboratory reference range. Dogs fed the HA‐GI diet had increased cardiac muscle carnitine esters (p = 0.014). Increased carnitine esters were also appreciated in cardiac muscle in all diets from Day 0 to 180 (p = 0.0001). On Day 180 mixed‐breed hounds had increased skeletal total carnitine (p < 0.001) compared to all time points and breeds. This study observed no correlation between plasma, whole blood, skeletal and cardiac muscle taurine concentrations but noted some effects between time, breed and diet. [ABSTRACT FROM AUTHOR]

Simple Summary: Neoadjuvant chemotherapy (NACT) is a pivotal treatment for breast cancer (BC). However, the success of NACT is uncertain and dependent on BC subtype. A valuable tool to identify biomarkers of the response to chemotherapy is metabolomics. We used plasma from BC patients together with clinical data to verify whether certain metabolites present before NACT can predict the responsiveness of the patient. Liquid chromatography–mass spectrometry and untargeted metabolomic analysis were performed. A statistical model was used to predict the response to NACT for samples separated into two sets: training and validation. The results showed 95.4%/93.3% sensitivity, 94.6%/94.7% accuracy, and 91.6%/100.0% specificity for each set, respectively. The compounds correctly classified 94.9% of resistant and 93.7% of sensitive females. The identified metabolites can be considered together with clinical data, allowing for the development of precision medicine strategies that lead to better treatment choices. Background: Neoadjuvant chemotherapy (NACT) has arisen as a treatment option for breast cancer (BC). However, the response to NACT is still unpredictable and dependent on cancer subtype. Metabolomics is a tool for predicting biomarkers and chemotherapy response. We used plasma to verify metabolomic alterations in BC before NACT, relating to clinical data. Methods: Liquid chromatography coupled to mass spectrometry (LC-MS) was performed on pre-NACT plasma from patients with BC (n = 75). After data filtering, an SVM model for classification was built and validated with 75%/25% of the data, respectively. Results: The model composed of 19 identified metabolites effectively predicted NACT response for training/validation sets with high sensitivity (95.4%/93.3%), specificity (91.6%/100.0%), and accuracy (94.6%/94.7%). In both sets, the panel correctly classified 95% of resistant and 94% of sensitive females. Most compounds identified by the model were lipids and amino acids and revealed pathway alterations related to chemoresistance. Conclusion: We developed a model for predicting patient response to NACT. These metabolite panels allow clinical gain by building precision medicine strategies based on tumor stratification. [ABSTRACT FROM AUTHOR]

Simple Summary: Lung cancer is the leading cause of cancer-related death. While sex and age impact outcomes in non-small cell lung cancer (NSCLC), it is not clearly understood how these factors affect tumor biology. We believe that sex and age influence the distribution of genomic alterations in NSCLC and evaluated for differences in predictive and/or prognostic alterations in individuals with advanced NSCLC. Our study is the largest, to our knowledge, to evaluate and confirm that the genomic landscape in advanced NSCLC differs by both sex and age. Genomic mutations impact non-small cell lung cancer (NSCLC) biology. The influence of sex and age on the distribution of these alterations is unclear. We analyzed circulating-tumor DNA from individuals with advanced NSCLC from March 2018 to October 2020. EGFR, KRAS, ALK, ROS1, BRAF, NTRK, ERBB2, RET, MET, PIK3CA, STK11, and TP53 alterations were assessed. We evaluated the differences by sex and age (<70 and ≥70) using Fisher's exact test. Of the 34,277 samples, 30,790 (89.83%) had a detectable mutation and 19,923 (58.12%) had an alteration of interest. The median age of the ctDNA positive population was 69 (18–102), 16,756 (54.42%) were female, and 28,835 (93.65%) had adenocarcinoma. Females had more alterations in all the assessed EGFR mutations, KRAS G12C, and ERBB2 ex20 ins. Males had higher numbers of MET amp and alterations in STK11 and TP53. Patients <70 years were more likely to have alterations in EGFR exon 19 del/exon 20 ins/T790M, KRAS G12C/D, ALK, ROS1, BRAF V600E, ERBB2 Ex20ins, MET amp, STK11, and TP53. Individuals ≥70 years were more likely to have alterations in EGFR L861Q, MET exon 14 skipping, and PIK3CA. We provided evidence of sex- and age-associated differences in the distribution of genomic alterations in individuals with advanced NSCLC. [ABSTRACT FROM AUTHOR]

Objective: This study aimed to evaluate the relationship between plasma soluble ST2 (sST2) levels 24 h after extracorporeal membrane oxygenation (ECMO) initiation and continuous renal replacement therapy (CRRT) in patients receiving venoarterial ECMO (V-A ECMO) support. Methods and Results: Data of patients who received ECMO support for postcardiotomy cardiogenic shock between January 2017 and July 2019 were retrospectively collected from Beijing Anzhen Hospital, Capital Medical University. Ultimately, 116 patients were included in the present study for analysis. The concentration of sST2 was determined by enzyme-linked immunosorbent assay (ELISA). The log10 sST2 levels were higher in patients undergoing CRRT than those who did not (6.06 vs. 6.22, p = 0.019). Patients undergoing CRRT had a lower survival rate than those who did not (32.8% vs. 67.3%, p < 0.001). In the univariate logistic regression analysis, sST2, HCO3-, lactate, and creatinine levels 24 h after ECMO initiation were related to CRRT (p < 0.05). In the multivariate logistic regression analysis, HCO3- and sST2 were identified as independent risk factors for CRRT use in patients undergoing ECMO (p < 0.05). The area under receiver operator characteristic curve (AUC) for sST2 and HCO3- together was 0.72 (95% confidence interval (CI), 0.79–0.91), which was better than those of sST2 or HCO3- alone (0.63 vs. 0.67). Conclusions: sST2 and HCO3−levels at 24 h after ECMO initiation were associated with CRRT and could predict CRRT use in postcardiotomy cardiogenic shock patients undergoing ECMO. [ABSTRACT FROM AUTHOR]

Introduction: Ethnic and gender differences in plasma lipid composition have been widely reported among the general population, but there are scarce data on athletes. Aim: To assess ethnic and gender differences in lipid profile across a large cohort of Olympic athletes practicing different sport disciplines Methods: We enrolled 1165 Olympic athletes divided into power, endurance, and mixed disciplines according to European Society of Cardiology classification. Sixty-two (5.3%) were Afro-Caribbean. Body composition and fat mass percentage were measured. Blood samples were collected and lipid profile was investigated. Results: Compared to Caucasians, Afro-Caribbeans had better lipid profile characterized by lower LDL (90 ± 25 mg/dL vs. 97.1 ± 26.2 mg/dL, p = 0.032) lower LDL/HDL ratio (1.39 ± 0.5 vs. 1.58 ± 0.6, p = 0.012), lower non-HDL-cholesterol (102.5 ± 27.4 mg/dL vs. 111.5 ± 28.5 mg/dL, p = 0.015) and lower TC/HDL (2.59 ± 0.6 vs. 2.82 ± 0.7, p = 0.010). Female Afro-Caribbeans showed lower TG/HDL ratio (p = 0.045) and TC/HDL ratio (p = 0.028), due to higher HDL (p = 0.005) compared to male Afro-Caribbeans. In Caucasian athletes, females showed even more evident differences with lower TC, LDL, and higher HDL with subsequent lower ratios compared to men. Moreover, endurance Caucasian athletes had lower LDL (p = 0.003) and TG (p = 0.017) plasmatic levels and higher HDL levels compared to non-endurance Caucasian athletes (p< 0.0001) Conclusions: Ethnicity and gender have a significant influence on plasmatic lipid balance in elite athletes and Afro-Caribbeans have favorable lipid profiles compared to Caucasians. Moreover, endurance sports, particularly in Caucasian athletes, are associated with better lipid profile compared to other type of sports. [ABSTRACT FROM AUTHOR]

Objectives: The adult‐onset focal dystonias are characterized by over‐active muscles leading to abnormal movements. For most cases, the etiology and pathogenesis remain unknown. In the current study, unbiased proteomics methods were used to identify potential changes in blood plasma proteins. Methods: A large‐scale unbiased proteomics screen was used to compare proteins (N = 6,345) in blood plasma of normal healthy controls (N = 49) with adult‐onset focal dystonia (N = 143) consisting of specific subpopulations of cervical dystonia (N = 45), laryngeal dystonia (N = 49), and blepharospasm (N = 49). Pathway analyses were conducted to identify relevant biological pathways. Finally, protein changes were used to build a prediction model for dystonia. Results: After correction for multiple comparisons, 15 proteins were associated with adult‐onset focal dystonia. Subgroup analyses revealed some proteins were shared across the dystonia subgroups while others were unique to 1 subgroup. The top biological pathways involved changes in the immune system, metal ion transport, and reactive oxygen species. A 4‐protein model showed high accuracy in discriminating control individuals from dystonia cases [average area under the curve (AUC) = 0.89]. Interpretation: These studies provide novel insights into the etiopathogenesis of dystonia, as well as novel potential biomarkers. ANN NEUROL 2024;96:110–120 [ABSTRACT FROM AUTHOR]

This study was conducted to investigate the effect of partial replacement of the crud protein of the concentrated feed mixture by Moringa oleifera leaves (MOL) on growth performance, hematological profile, and some blood plasma constituents of growing lambs. Twenty-four healthy male Farafra lambs (5 and 6 months old and 30 ± 0.50 kg body weight) were randomly divided into four groups. The first group was fed on 60% of the concentrated feed mixture (CFM) and 40% wheat straw as DM and served as a control. The other 3 treatment groups (10 MOL, 20 MOL, and 30 MOL), partially substituted the crud protein of the CFM at rates 10, 20, and 30% with Moringa oleifera leaves protein, respectively. The results revealed that dietary MOL had no significant effect on body weight gain, however they may adversely be affected by increasing the levels of MOL. The inclusion MOL at 20% in the diet of lambs increased (P <0.05) WBC’s count particularly neutrophils as compared with other groups. Also, MOL groups tended to be higher in RBC’s count and percentages of HG and HCT than control one. The plasma albumin was decreased (P˂0.05) with increasing levels of MOL in the diets of lambs. Also, the urea concentration and ALT values were lower (P˂0.05) with MOL supplementation at Levels of 20 and 30% than that in 10% group. It could be concluded that the partial substitution of concentrate feed with Moringa oleifera leaves up to 10 % growing lambs' diets can improve growth performance, blood hematology, and plasma constituents, and exert protective effects in the liver without inducing adverse effects on animal health. [ABSTRACT FROM AUTHOR]

Following an application from Cárnicas Joselito S.A. pursuant to Article 14 of Regulation (EC) No 1924/2006 via the Competent Authority of Spain, the Panel on Nutrition, Novel Foods and Food Allergens (NDA) was asked to deliver an opinion on the scientific substantiation of a health claim related to 'Joselito ham increases antioxidant substances in the body, reduces blood pressure and plasma triglycerides, decreases oxidative stress and prevents effect in diseases related to the cardiovascular and intestinal systems'. The scope of the application was proposed to fall under a health claim referring to disease risk reduction. The food constituent that is the subject of the health claim is Joselito, an Iberian ham characterised by a high content of oleic acid. The Panel considers that the food is sufficiently characterised. The Panel considers that lowering of LDL‐cholesterol concentration and blood pressure is a beneficial effect by decreasing the risk of coronary heart disease. Upon a request from EFSA, the applicant identified one human intervention study as being pertinent to the claim. However, due to methodological limitations, the Panel considers that no conclusions can be drawn from this study for the scientific substantiation of the claim. The Panel notes that no human intervention studies from which conclusions could be drawn for the scientific substantiation of the claim were provided by the applicant. The Panel concludes that a cause and effect relationship has not been established between the intake of Joselito® ham and the reduction of LDL‐cholesterol concentration or blood pressure. [ABSTRACT FROM AUTHOR]

Background and Objectives: An increasing number of blood operators around the world, including those in Canada, have removed time‐based deferral periods for gay, bisexual and other men who have sex with men and replaced them with sexual behaviour‐based questions for all donors. While this marks a significant shift in screening approach, what remains unclear is how members of two‐spirit, lesbian, gay, bisexual, transgender and queer (2S/LGBTQ+) communities view blood operators' initiatives to be more inclusive. As such, this study was conducted to assess the awareness of donor screening changes and other initiatives among members of 2S/LGBTQ+ communities and to explore their recommendations for blood operators' work with these communities. Materials and Methods: Semi‐structured qualitative interviews (n = 15) were conducted with 2S/LGBTQ+ people across Canada. Data were analysed using open inductive coding methods. Results: Reported here are the key results on recommendations for blood operators. Three themes were identified from the data: (1) the need for increased communications with 2S/LGBTQ+ communities surrounding changes to donor policies and guidelines; (2) the need for trans‐inclusive policy and procedures; and (3) the need for culturally responsive and equity‐informed staff training at donor centres. Conclusion: Results suggest that blood operators should consider 2S/LGTBQ+ communities when developing blood and plasma donation policies, screening procedures and staff training. Increased consultation with these communities is desired, and further research specific to the experiences of transgender blood donors is needed. [ABSTRACT FROM AUTHOR]