Serum Samples Research Articles

Construction of pseudotyped human coronaviruses and detection of pre-existing antibodies in the human population

In order to clarify the pre-exist immunity background of different human coronaviruses, this study investigated the positive rate of spike (S) protein antibodies of seven human coronaviruses (HCoV), including HCoV- severe acute respiratory syndrome (SARS) −associated coronavirus (SARS-CoV-1), severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Middle East respiratory syndrome coronavirus (MERS-CoV), HCoV-229E, HCoV-NL63, HCoV-HKU1 and HCoV-OC43, before and after the Coronavirus Disease 2019 (COVID-19) outbreak. We utilized pseudotyped virus-based neutralization assays (PBNA) or enzyme-linked immunosorbent assays (ELISA) to detect antibody levels against human coronaviruses in serum samples collected in 2009–2010 and 2023. The PBNA results showed that neutralizing antibodies against SARS-CoV-1 and the MERS-CoV were negative. In the serum samples from 2009 to 2010, neutralizing antibodies against SARS-CoV-2 (D614G) were negative, whereas in the serum samples from 2023, 73 samples (73 %) showed neutralizing reactions with the SARS-CoV-2 D614G strain, 96 samples (96 %) with the BA.5 strain, and 91 samples (91 %) with the BF.7 strain. Among pre-COVID-19 samples, 33 % (33/100) showed neutralizing reactions with HCoV-229E and 63 % (63/100) with HCoV-NL63. Among post-COVID-19 samples, 50 % (50/100) showed neutralizing reactions with HCoV-229E and 49 % (49/100) with HCoV-NL63. Due to the different receptors of alpha coronavirus genus compared to other beta coronavirus genus, neutralizing antibodies against HCoV-OC43 and HCoV-HKU1 virus cannot be detected by constructing corresponding pseudotyped virus. Binding antibodies against HCoV-OC43 and HCoV-HKU1 virus were detected using ELISA. The results revealed that among pre-COVID-19 samples, 83 % (83/100) and 45 % (45/100) had binding activity with HCoV-OC43 and HCoV-HKU1, respectively. Among post-COVID-19 samples, 100 % (100/100) and 81 % (81/100) had binding activity with HCoV-OC43 and HCoV-HKU1, respectively.

Untargeted Metabolomic Analyses of Body Fluids to Differentiate TBI DOC and NTBI DOC.

To investigate the metabolomic differences between Traumatic brain injury (TBI) disorder of consciousness (DOC) patients and non-traumatic brain injury (NTBI) DOC patients by using cerebrospinal fluid (CSF), serum and urine samples beneficial to understand the pathological mechanism differences between the two etiologies, provide potential clues for the subsequent treatment and prognosis, and investigate the metabolome differences and similarities between TBI and NTBI among three different body fluids. In total, 24 TBI DOC subjects and 29 NTBI DOC subjects were enrolled. CSF, serum and urine samples from TBI DOC and NTBI DOC patients were collected and analyzed by performing UPLC-MS. The statistical methods and pathway analyses were applied to discover potential biomarkers and altered metabolic functions. When comparing TBI DOC and NTBI DOC, 36, 31 and 52 differential metabolites were obtained in CSF, serum and urine, respectively. The functional analysis of differential metabolites obtained in CSF, serum and urine were all related to amino acid metabolism. Except for amino acid metabolism, metabolic biomarkers in CSF, serum and urine mainly focus on central function, cognitive function, necrosis and apoptosis and neurological function, respectively. In CSF, the highest AUC was 0.864 (Isoproturon) and 0.816 (Proline betaine). Then, the AUC of NFurfurylformamide in serum was 0.941, while the AUC of Dihydronepetalactone and Doxepin N-oxide glucuronide were 1.0 in urine. CSF, serum and urine metabolomic analyses could differentiate TBI DOC from NTBI DOC and functional analyses showed a metabolic change difference between TBI DOC and NTBI DOC.

Adjuvant therapy with zinc supplementation; anti-inflammatory and anti-oxidative role in multiple myeloma patients receiving autologous hematopoietic stem cell transplantation: a randomized controlled clinical trial.

Multiple myeloma (MM) patients are often accompanied by heightened levels of oxidative stress, even following bone marrow transplantation. Trace mineral supplements have been found to regulate and inhibit the activity of oxidative radicals and inflammatory factors, which are involved in the pathogenesis of MM. The study sought to evaluate the effectiveness of the supplementation by analyzing changes in oxidative, anti-oxidative, and inflammation markers. Patients were randomly assigned to a zinc or placebo group, with the former receiving 30mg of zinc or placebo tablets daily for 1month. Blood samples were collected from the patients on the day of transplantation, 15days, and 30days post-transplantation. Real-time PCR was employed to measure the expression of oxidative/antioxidative genes. Furthermore, the protein level of oxidative markers in serum samples was assessed. Finally, serum TNF-α concentrations were measured using the ELISA technique. The expression levels of SOD1, SOD2, and NRF2 genes were significantly higher on days 15 and 30 compared to the control group (P < 0.05), with a greater increase on day 30 (P < 0.05). Conversely, the expression levels of Keap1 and NOX2 genes were lower on day 30 than those of the control group (P < 0.05), with a further decrease from day 15 to day 30 (P < 0.05). The experimental group exhibited a notable reduction in TNF-α cytokine levels on day 30 compared to the control and placebo groups (P < 0.05). All findings were coordinated according to the nutritional questionnaire. Our findings suggest a potential benefit of zinc supplementation in managing the adverse effects of chemotherapy in MM patients, warranting further investigation.

Candidate Biomarkers For Response To Treatment In Psoriatic Disease.

To determine whether biologic therapy alters serum CXCL10, MMP3, S100A8, ACP5, and CCL2 levels in psoriatic arthritis (PsA) and psoriasis (PsC) patients, and whether baseline levels of these proteins predict response to treatment for PsA. We included PsA patients on TNF inhibitors (TNFi), Interleukin-17 inhibitors (IL-17i), methotrexate (MTX), and untreated with bDMARDs or csDMARDs (untreated) and PsC patients on bDMARDs and matched untreated. Serum samples at baseline and 3-6 months follow-up were retrieved from the biobank. Protein levels were quantified using a Luminex multiplex assay. We compared follow-up vs baseline protein levels within groups and change in levels between groups. For the predictive potential of the biomarkers, we developed logistic regression classification models. Response to treatment was defined as: 1. Achieving low disease activity or remission (according to DAPSA), 2. 75% reduction in PASI, and 3. 50% reduction in actively inflamed joint count. In PsA, TNFi reduced serum levels of all five proteins, IL-17i increased ACP5 and CCL2, and MTX reduced MMP3. Changes in MMP3 and S100A8 levels were significantly different between untreated PsA and matched biologic-treated PsA (p<0.05). There were no significant differences between treated or untreated PsC patients. Baseline levels of CXCL10, MMP3, S100A8, and ACP5 had good predictive value (AUC>0.8) for response to biologics in PsA patients. Biologics treatment and MTX affect serum CXCL10, MMP3, S100A8, ACP5, and CCL2 levels in PsA patients. MMP3, S100A8, ACP5, and CXCL10 have potential use as serum biomarkers to predict response to treatment for PsA.

Differential serum levels of CACNA1C, circadian rhythm and stress response molecules in subjects with bipolar disorder: Associations with genetic and clinical factors

BackgroundMany patients with bipolar disorder (BD) do not respond to or have difficulties tolerating lithium and/or other mood stabilizing agents. There is a need for personalized treatments based on biomarkers in guiding treatment options. The calcium voltage-gated channel CACNA1C is a promising candidate for developing personalized treatments. CACNA1C is implicated in BD by genome-wide association studies and several lines of evidence suggest that targeting L-type calcium channels could be an effective treatment strategy. However, before such individualized treatments can be pursued, biomarkers predicting treatment response need to be developed. MethodsAs a first step in testing the hypothesis that CACNA1C genotype is associated with serum levels of CACNA1C, we conducted ELISA measures on serum samples from 100 subjects with BD and 100 control subjects. ResultsWe observed significantly higher CACNA1C (p < 0.01) protein levels in subjects with BD. The risk single nucleotide polymorpshism (SNP) (rs11062170) showed functional significance as subjects homozygous for the risk allele (CC) had significantly greater CACNA1C protein levels compared to subjects with one (p = 0.013) or no copies (p = 0.009). We observed higher somatostatin (SST) (p < 0.003) protein levels and lower levels of the clock protein aryl hydrocarbon receptor nuclear translocator-like (ARTNL) (p < 0.03) and stress signaling factor corticotrophin releasing hormone (CRH) (p < 0.001) in BD. SST and period 2 (PER2) protein levels were associated with both alcohol dependence and lithium response. ConclusionsOur findings represent the first evidence for increased serum levels of CACNA1C in BD. Along with altered levels of SST, ARNTL, and CRH our findings suggest CACNA1C is associated with circadian rhythm and stress response disturbances in BD.

Metabolomics study of lipid lowering effect and lysophospholipids regulation by Alismatis rhizoma and processed forms in hyperlipidemia mice

Alismatis rhizoma (AR) is a traditional herb used for its lipid-regulating properties. Its processed forms, Salt-processed AR (SAR) and bran-processed AR (BAR), are widely used. This study investigates the lipid-lowering effects of AR and its processed forms in hyperlipidemic mice, with a focus on lysophospholipid regulation. Three types of serum lysophospholipids were characterized using UHPLC-QTOF-MS/MS, and their metabolic changes were analyzed with multivariate statistical statistics. The results showed that AR effectively reduced total cholesterol (TC), while SAR and BAR excelled in lowering low-density lipoprotein cholesterol (LDL-C). BAR demonstrated superior effects on the TC/high-density lipoprotein-cholesterol (HDL-C) ratio, atherogenic index (AI), and protecting kidney function, making it the most effective processed forms. Additionally, a total of 216 lysophospholipids, including 153 lysophosphatidylcholines (Lyso-PCs), 49 lysophosphatidylethanolamines (Lyso-PEs), and 14 lysophosphatidylserines (Lyso-PSs), were identified in serum samples. Metabolomics analysis revealed 102 differential lysophospholipids associated with hyperlipidemia, among which 29, 21, and 22 were significantly (VIP>1.0, P<0.05) regulated by AR, SAR, and BAR, respectively. AR showed the most comprehensive regulation of lysophospholipids, increasing unsaturated Lyso-PCs and decreasing Lyso-PEs and Lyso-PSs, which might reduce inflammation and improve cardiovascular health. This study is the first to comprehensively compare the lipid-lowering effects of AR and its processed forms, highlighting their role in modulating lysophospholipid metabolism in hyperlipidemia.

Antibodies Against ZSCAN1 in Pediatric and Adult Patients With Non-Paraneoplastic ROHHAD Syndrome.

To report the association of zinc finger and SCAN domain containing 1 antibodies (ZSCAN1-abs) with rapid-onset obesity, hypothalamic dysfunction, hypoventilation, and autonomic dysregulation (ROHHAD) syndrome in patients without tumor. Patients with symptoms compatible with ROHHAD syndrome but without an associated tumor were selected from our database. Serum and CSF samples were examined for the presence of ZSCAN1-abs by an in-house cell-based assay. In addition, samples from 149 patients with several inflammatory and noninflammatory disorders and 50 healthy participants served as controls. Thirteen patients with ROHHAD syndrome were identified. Of these, we had paired serum/CSF samples from 6 patients and only serum from the other 7. Five of 6 patients (83.3%) with paired serum/CSF (4 children, 1 adult) had ZSCAN-abs only in CSF and 1 had antibodies in serum and CSF. ZSCAN1-abs were not detected in the remaining 7 patients with ROHHAD with only serum available or in any of the 199 control samples. Patients with ROHHAD syndrome should be investigated for the presence of ZSCAN1-abs in CSF. The antibodies do not necessarily predict the presence of a tumor. The detection of ZSCAN1-abs in an adult patient suggests that this condition also occurs beyond the pediatric age.

Metabolic Choreography of Energy Substrates During DCD Heart Perfusion.

The number of patients waiting for heart transplant far exceeds the number of hearts available. Donation after circulatory death (DCD) combined with machine perfusion can increase the number of transplantable hearts by as much as 48%. Emerging studies also suggest machine perfusion could enable allograft "reconditioning" to optimize outcomes. However, a detailed understanding of the energetic substrates and metabolic changes during perfusion is lacking. Metabolites were analyzed using 1-dimensional 1H and 2-dimensional 13C-1H heteronuclear spectrum quantum correlation nuclear magnetic resonance spectroscopy on serial perfusate samples (N = 98) from 32 DCD hearts that were successfully transplanted. Wilcoxon signed-rank and Kruskal-Wallis tests were used to test for significant differences in metabolite resonances during perfusion and network analysis was used to uncover altered metabolic pathways. Metabolite differences were observed comparing baseline perfusate to samples from hearts at time points 1-2, 3-4, and 5-6 h of perfusion and all pairwise combinations. Among the most significant changes observed were a steady decrease in fatty acids and succinate and an increase in amino acids, especially alanine, glutamine, and glycine. This core set of metabolites was also altered in a DCD porcine model perfused with a nonblood-based perfusate. Temporal metabolic changes were identified during ex vivo perfusion of DCD hearts. Fatty acids, which are normally the predominant myocardial energy source, are rapidly depleted, while amino acids such as alanine, glutamine, and glycine increase. We also noted depletion of ketone, β-hydroxybutyric acid, which is known to have cardioprotective properties. Collectively, these results suggest a shift in energy substrates and provide a basis to design optimal preservation techniques during perfusion.

Myositis -specific and -associated antibodies in neurological disorders - A retrospective study of 727 patients

ObjectiveMyositis-specific antibodies (MSAs) and myositis-associated antibodies (MAAs) are assessed in clinical neurology, serving as a non-invasive tool for the differential diagnosis of autoimmune myopathies. However, the presence of MSAs and MAAs in neurological disorders remains uncertain. MethodsRetrospective analysis was conducted on 878 serum samples from the neurological laboratory of the University Hospital Tübingen, Germany. The EUROLINE Myositis Profil 3 (IgG) Line Blot was used for antibody evaluation (anti-Mi2, -Ku, -PM-Scl100, -PM-Scl75, −Jo1, -SRP, -PL7, -PL12, −EJ, -OJ, and -Ro52). Samples were categorized into 19 disease groups, with consideration for myositis-linked and non-myositis-linked diseases. Then, the distribution of positive findings and the concurrent presence of more than one MAA/MSA were analyzed. ResultsAmong 727 included line blots, 84 could be assigned to myositis-linked diseases (thereof 44 positive for MAA/MSA). MAA and MSA taken together were more frequently positive for the main group of myositis-linked disease (52.4 %) compared to the non-myositis-linked group (14.6 %, overall specificity 85.4 %). However, individual antibodies were specific, ranging above 97.5 %. False positive antibody results can also occur in neurological differential diagnoses such as muscle dystrophy or cramp fasciculation syndrome. Furthermore, the concurrent presence of more than one MAA/MSA does not show a significant association with the presence of a myositis-linked disease for antibody-positive samples (p = 0.136). DiscussionTesting MSA and MAA simultaneously may not be suitable as a primary screening method for myositis-linked diseases in clinical neurological groups. However, MSAs and MAAs may offer valuable diagnostic support, particularly in cases where myositis is strongly considered.

Perfluoroalkyl substances exposure and the risk of breast cancer: A nested case-control study in Jinchang Cohort

BackgroundAs persistent organic pollutants (POPs), perfluoroalkyl substances (PFAS) may potentially impact human health. Our study aimed to investigate the prospective association between PFAS exposure and the incidence risk of breast cancer in females. MethodsBy fully following the Jinchang Cohort after a decade, we conducted this nested case-control study with 135 incidence cases of breast cancer (BC) and 540 bias-paired controls. The PFAS levels were tested by baseline serum samples. Conditional logistic regression and a restricted cubic spline model were employed to investigate the BC incidence risks and the dose-response associated with single PFAS component exposure. Furthermore, the Quantile g-computation model (Qgc), random forest model (RFM), and bayesian kernel machine regression models (BKMR) were integrated to estimate the mixed effects of PFAS exposure on the incidence risk of BC. ResultsExposures to specific PFAS components were positively associated with an increased incidence risk of breast cancer. By grouping the study population into different baseline menopausal statuses, PFHxS, PFNA, PFBA, PFUdA, PFOS, and PFDA demonstrated a similarly positive correlation with BC incidence risks. However, the increased incidence risks of BC associated with PFOA, PFOS, PFUdA, and 9CL-PF3ONS exposure were exclusively found in the premenopausal population. Both BKMR and Qgc revealed that exposure to mixed PFAS was associated with an increased risk of breast cancer, with Qgc specifically indicating an odds ratio (OR) of 2.21 (95% CI: 1.53, 3.19). Random forests showed that PFBA, PFOS, PFHxS, and PFDA emerged as predominant factors potentially influencing breast cancer incidence. ConclusionOur findings suggest a strong association between PFAS exposure and the incidence of breast cancer. Premenopausal women should exercise more caution regarding PFAS exposure.

Plasmonic sensing platform for C-reactive protein recognition via synthetic receptors decorated on graphene oxide and gold nanoparticles

C-reactive protein (CRP) level provides important information about the health status of the individual in predicting many diseases such as cardiovascular, chronic inflammatory, and neurodegenerative diseases. Therefore, determining CRP levels is important for correct health intervention and treatment follow-up. For the selective detection of CRP, we developed a (GO/Au-MIP) SPR sensor containing CRP-imprinted polymer modified with graphene oxide and gold nanoparticles. To prove the enhanced sensitivity of the sensor resulting from the presence of graphene oxide (GO) and gold nanoparticles (AuNPs), a CRP-imprinted (MIP) SPR sensor was prepared using the same method, excluding the incorporation of GO and AuNPs. In addition, a non-imprinted GO/Au-NIP SPR sensor was also prepared to evaluate the imprinting efficiency. In detecting CRP in PBS buffer, the GO/Au-MIP SPR sensor exhibited linearity in the concentration ranges of 0.1–2 ppm (R2 = 0.9721) and 5–100 ppm (R2 = 0.9740). The detection limit of the prepared sensor was calculated as 0.0082 ppm. In the study, the imprinting process efficiency was also evaluated by calculating the imprinting factor value (I.F=13.84). In the selectivity studies, the GO/Au-MIP SPR sensor was determined to be 9.23 times more selective against CRP protein than bovine serum albumin and 29.53 times more selective than hemoglobin. When the repeatability of the GO/Au-MIP SPR sensor was examined, it was determined that the GO/Au-MIP SPR sensor was able to detect CRP without any deterioration in performance in five consecutive reuses (RSD<1.5). Finally, CRP detection studies from the serum and urine solutions, which were selected as real samples, were carried out by the GO/Au-MIP SPR sensor to evaluate the matrix effect. CRP spiked serum sample was analyzed by the other standard method to validate the analytical results using CRP-Analyser.

Fabrication of rGO-decorated hBNNS hybrid nanocomposite via organic-inorganic interfacial chemistry for enhanced electrocatalytic detection of carcinoembryonic antigen.

Organic-inorganic hybrid nanocomposites (OIHN), with tailored surface chemistry, offer ultra-sensitive architecture capable of detecting ultra-low concentrations of target analytes with precision. In the present work, a novel nano-biosensor was fabricated, acquainting dynamic synergy of reduced graphene oxide (rGO) decorated hexagonal boron nitride nanosheets (hBNNS) for detection of carcinoembryonic antigen (CEA). Extensive spectroscopic and microscopic analyses confirmed the successful hydrothermal synthesis of cross-linked rGO-hBNNS nanocomposite. Uniform micro-electrodes of rGO-hBNNS onto pre-hydrolyzed ITO were obtained via electrophoretic deposition (EPD) technique at low DC potential (15V). Optimization of antibody incubation time, pH of supporting electrolyte, and immunoelectrode preparation was thoroughly investigated to enhance nano-biosensing efficacy. rGO-modified hBNNS demonstrated 29% boost in electrochemical performance over bare hBNNS, signifying remarkable electro-catalytic activity of nano-biosensor. The presence of multifunctional groups on the interface facilitated stable crosslinking chemistry, increased immobilization density, and enabled site-specific anchoring of Anti-CEA, resulting in improved binding affinity. The nano-biosensor demonstrated a remarkably low limit of detection of 5.47pg/mL (R2 = 0.99963), indicating exceptional sensitivity and accuracy in detecting CEA concentrations from 0 to 50ng/mL. The clinical evaluation confirmed its exceptional shelf life, minimal cross-reactivity, and robust recovery rates in human serum samples, thereby unraveling the potential for early, highly sensitive, and reliable CEA detection.

Comparison of Serum Vitamins C, D, and E in Iraqi Colorectal Cancer Patients with and without Non-Alcoholic Fatty Liver Disease to Healthy Individuals

Abstract Depending on their protective properties against different cases of Colorectal Cancer (CRC), vitamins C, D, and E are the main focus of this research. CRC is one of the global public health concerns. 30 healthy individuals provided serum samples, whereas the group of CRC patients was divided into three, totaling 90 individuals. Group I consisted of 30 newly diagnosed cases of CRC. Group II 30 consisted of consisted of 30 CRC patients who were administered three cycles of chemotherapy. Group III consisted of 30 diagnosed CRC patients who also have non-alcoholic fatty liver disease (NAFLD). The concentrations and groups of vitamins C, D, and E were evaluated using ELISA. The levels of Vitamin C were significantly lower (p &lt;0.0001) in individuals newly diagnosed with CRC, as compared to the control group. Patients who were recently diagnosed and treated for CRC and NAFLD exhibited comparable amounts of vitamin C. The level of vitamin E in control, newly diagnosed CRC and treated CRC patients displayed no significant differences, however comparing with newly diagnosed patients that suffered NAFLD, a significant decrease (p = 0.0014) in the level of vitamin E in CRC with NAFLD group was. The levels of Vitamin D in all groups of patients with colorectal cancer were significantly lower than those in the control group (p &lt;0.0001). Colon cancer patients exhibited significantly decreased levels of vitamin C and D compared to healthy individuals. Vitamins showed potential as diagnostic markers for distinguishing between groups. It is necessary to analyze these findings as the level of serum vitamins may have potential use in the prognosis of the CRC in patients.

Analytical performance evaluation of a new integrated clinical chemistry and immunoassay analyzer

BackgroundClinical laboratories perform a wide range of tests that are used by healthcare professionals to guide medical decision making. Use of automated analyzers in the clinical laboratory can improve patient care by not only reducing the turn-around-time (TAT) of results but also improving accuracy of the reported results by reducing human error. The aim of this study was to evaluate the performance characteristics of a new automated laboratory instrument, the Atellica® CI Analyzer, Model 1900, over a 3-month period in a European laboratory setting. MethodsAnalytical performance of 17 analytes (13 chemistry and four immunochemistry) was assessed by evaluating repeatability and within-laboratory precision using anonymized remnant serum samples. Method comparison studies were performed on the Atellica CI Analyzer and the Roche cobas® 6000. ResultsExcellent precision was observed with coefficients of variation (CVs) less than 2% for repeatability and less than 3% within-laboratory imprecision for most analytes. Comparison of select assays with the cobas 6000 system resulted in correlation coefficients ranging from 0.980 – 1.000. ConclusionThis is the first reported evaluation of the Atellica CI Analyzer in a clinical laboratory setting. The strong analytical performance of the Atellica CI Analyzer demonstrates that this instrument is suitable for routine clinical use.

Evaluation of immunological status in pregnant and aborted women with cytomegal infection in Thi-Qar Governorate

The current study was conducted on aborted and pregnant women suspected of being infected with cytomegalovirus in The-Qar Governorate. (90) Serum samples of pregnant and aborted women were examined using the Enzyme-linked Immunosorbent assay(ELISA). through the examination, (50) sample of women infected with CMV virus were found, with rate of 55.55%, where it was found that (43) sample of them carried the IgG antibody, with a rate of 48.88%, and(5) sample with the IgM immunoglobulin antibody, with a rate of (5.5%), and (2) sample of Igm+IgG, with a rate of (2.2%). Through the study of blood parameters that was conducted on (40) blood samples of women infected with the virus, compared to (10) blood samples of uninfected women there was a decrease in the blood hemoglobin rate (Hb)(11.54%) and compressed blood volume by (0.35) compared to the control group, three were no significant difference(p&gt;0.05) for the total number of white blood cells. When conducting a differential of white blood cells for infected women it was found that there was an increase in the percentage of lymphocytes by (40.50), While no significant differences were observed (p&gt;0.05) in the percentage of monocytes,neutrophils eosinophils, and basophils. As for the phagocytosis factor, an increase in the phagocytosis factor was recorded for infected women compared to the control group, as the phagocytosis factor for infected women reached 12-60% and for the control group 47.10%.

The trajectory of vesicular proteomic signatures from HBV-HCC by chitosan-magnetic bead-based separation and DIA-proteomic analysis.

Hepatocellular carcinoma (HCC) is a prevalent primary liver cancer often associated with chronic hepatitis B virus infection (CHB) and liver cirrhosis (LC), underscoring the critical need for biomarker discovery to improve patient outcomes. Emerging as a promising avenue for biomarker development, proteomic technology leveraging liquid biopsy from small extracellular vesicles (sEV) offers new insights. Here, we evaluated various methods for sEV isolation and identified polysaccharide chitosan (CS) as an optimal approach. Subsequently, we employed optimized CS-based magnetic beads (Mag-CS) for sEV separation from serum samples of healthy controls, CHB, LC, and HBV-HCC patients. Leveraging data-independent acquisition mass spectrometry coupled with machine learning, we uncovered potential vesicular protein biomarker signatures (KNG1, F11, KLKB1, CAPNS1, CDH1, CPN2, NME2) capable of distinguishing HBV-HCC from CHB, LC, and non-HCC conditions. Collectively, our findings highlight the utility of Mag-CS-based sEV isolation for identifying early detection biomarkers in HBV-HCC.

Unveiling the seroprevalence of human papillomavirus in Guangdong, China: Implications for vaccination strategies.

Seroepidemiological characteristics of human papillomavirus (HPV) in community residents reflect natural infection and can guide the reform of vaccination programs. A population-based serological survey was conducted in Guangdong Province. Serum anti-HPV IgG antibody levels were determined by an ELISA. Neutralizing antibodies against HPV6, 11, 16, and 18 were detected via a pseudovirus-based neutralization assay (PBNA). A total of 5122 serum samples were collected from community residents, including 1989 males and 3133 females, in three cities of Guangdong Province. The rate of HPV IgG antibody positivity in females was 5.39% (95% CI: 4.6-6.2), which was greater than that in males (2.36%; 95% CI: 1.7-3.1). HPV IgG antibodies were more frequently detected in females aged 51-60 years (11.30%; 95% CI: 7.6-16.0), whereas in males, the detection increased with age and reached 4.94% (95% CI: 2.8-6.9) in the group aged ≥71 years. The seropositivity of neutralizing antibodies against HPV6 and 11 was greater than that against HPV16 and 18. The serum neutralizing antibody titers in individuals who received three doses of a vaccine were 7- to 12-fold greater than those in individuals who did not receive the vaccine. The neutralizing antibody titers slightly decreased within 40 months and ranged from 0.038 to 0.057 log ED50 per month. A moderate consistency between the HPV ELISA and PBNA results was observed (Kappa score = 0.49, r = 0.249, 0.635, 0.382, and 0.466 for HPV6, 11, 16, and 18, respectively). The HPV seropositivity rate among healthy residents of Guangdong Province was found to be low among children and adolescents and to increase with age. The serum neutralizing antibody titers were significantly greater in the vaccine group than that in the control group, and this difference persisted over time, which indicated promising protection against HPV infection.

A "signal-off" anodic photoelectrochemical sensor based on ZnIn2S4/TiO2 heterojunction for dopamine detection

Dopamine (DA) is an important neurotransmitter. Abnormal levels of it in human body can increase the risk of many neurological diseases. Thus, developing a simple, sensitive detection method of DA is crucial. In this paper, we reported a "signal-off" anodic PEC sensor based on fluorine-doped tin oxide (FTO) glass modified ZnIn2S4/TiO2 heterojunction (ZnIn2S4/TiO2/FTO) for DA detection. The experimental results show that the ZnIn2S4/TiO2/FTO electrode prepared by two-step hydrothermal method has a good photocurrent response performance under visible light. After incubation with DA, the photocurrent response decreases significantly because DA can rapidly oxidizes to polydopamine (PDA) through the action of superoxide radical (·O2-) and hydroxyl radical (·OH) intermediate species, which are intermediates produced by the ZnIn2S4/TiO2/FTO electrode under visible light irradiation. The constructed PEC sensor has a good linear relationship in the concentration range from 0.5 to 1000.0 μM, and its detection limit is 0.253 μM. In addition, the results of the proposed PEC sensor in real serum samples are satisfactory. The PEC sensor provides a promising platform for DA detection, laying the foundation for future advances in disease diagnosis and prevention.

SERS and electrochemical dual-mode detection of miRNA-141 by using single Au@Ag nanowire as a new platform.

As biomarkers of cancer, the accurate and sensitive detection of microRNAs is of great significance. Therefore, we proposed a surface-enhanced Raman scattering (SERS)/electrochemical (EC) dual-mode nanosensor for sensitively detecting miRNA-141. The nanosensor uses Au@Ag nanowires as a novel SERS/EC sensing platform, which has the advantages of good biocompatibility, fast response, and high sensitivity. The dual-mode nanosensor can not only effectively overcome the problem of insufficient reliability of single signal, but also realize the amplification and stable output of the detection signal, to ensure the reliability and repeatability of miRNA detection. With this sensing strategy, the target miRNA-141 can be detected over a wide linear range (100 fM to 50 nM) (LOD of 18.4 fM for SERS and 16.0 fM for electrochemical methods). In addition, the process shows good selectivity and can distinguish miRNA-141 from other interfering miRNAs. The actual analysis of human serum samples also proves that our strategy has good reliability, repeatability, and has broad application prospects in the field of analysis and detection.

Prevalence of bluetongue virus disease in a small ruminant population in Kalat, Balochistan, Pakistan

Background and Aim: Bluetongue is a vector-borne, emerging disease that poses a severe threat to most domesticated animals. A cross-sectional study was conducted to estimate the prevalence of bluetongue virus (BTV) disease in apparently healthy sheep and goats in Kalat, Balochistan. Materials and Methods: A total of 270 serum samples (sheep: 150 and goat: 120) were collected and screened for the detection of anti-BTV antibodies using a competitive enzyme-linked immunosorbent assay. The data regarding different contributory risk factors were also collected using a predesigned questionnaire. Results: It revealed that overall, 27.4% (74/270, 95% confidence interval, χ2 = 1.71, p = 0.12) prevalence in both sheep and goat populations. The highest prevalence of 47% (32/68) was recorded in Surab city with the lowest prevalence of 15.49% (11/71) in the Manguchar area. In contrast, in Kalat 28.1% (9/32), Daan area 24% (12/50), and Marap area 22.44% (11/49), seropositivity was recorded. Upon sex bases, antibodies were almost equally found in both male 28.57% (8/28) and female 27.27% (66/242) animal populations. Moreover, all four breeds (Balochi, Khurasani, Lehri, and Rakhshani) were equally and potentially seropositive. The Khurasani breed was the most susceptible to 34.69% (17/49), followed by the Balochi breed, 45/145 (31%) seropositivity. The prevalence of BTV was 16.66% (1/6) in Rakhshani breed and 15.71% (11/70) in Lehri breed., Ticks were found in almost 21% of animals, while 93% of animals were reared on open grazing in rangelands. Conclusion: This study clearly indicates widespread BTV infection in small ruminants in the study area that may pose serious threats to livestock farming. Further extensive studies are recommended to study the prevalence of disease in different agroecological zones of the province. This also warns the high-ups to manage concrete efforts to eradicate and control the disease in the area. Keywords: antibodies, Balochistan, bluetongue virus, competitive enzyme-linked immunosorbent assay, Kalat.