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Background: Platelet-rich fibrin (PRF) is used to prepare "sticky bone" by combining it with bone graft material. The present study investigated the ability of different bone grafts to absorb growth factors from the PRF and release them over time. Materials and Methods: Human blood was collected from 10 healthy volunteers for liquid PRF preparation. Bovine bone, allograft (mineralized and demineralized), and synthetic bone were each mixed with the PRF to prepare a sticky bone. All sticky bone samples were incubated for up to 4 days. The absorption and release pattern kinetics of two selective growth factors within the PRF--platelet-derived growth factor (PDGF) and bone morphogenetic protein-2 (BMP-2)--were quantified with immunofluorescence staining and enzyme-linked immunoassay (ELISA) testing. Results: All bone graft materials adsorbed the examined growth factors from the PRF. ß-TCP showed the highest adsorption levels, followed by the xenograft, and the allografts showed the lowest adsorption levels. Furthermore, PDGF showed a fast-release pattern from the grafts, whereas BMP-2 was released at a later stage. Similar to the adsorption pattern, the ß-TCP and xenograft were better able to sustain the release of the PRF growth factors from the graft than the allografts. Conclusions: The adsorption of PDGF and BMP-2 differ between graft materials, with superior results for ß-TCP, followed by xenograft, then allograft materials. [ABSTRACT FROM AUTHOR]

Intestinal transplantation is the definitive treatment for intestinal failure. However, tissue rejection and graft-versus-host disease are relatively common complications, necessitating aggressive immunosuppression that can itself pose further complications. Tracking intraluminal markers in ileal effluent from standard ileostomies may present a noninvasive and sensitive way to detect developing pathology within the intestinal graft. This would be an improvement compared to current assessments, which are limited by poor sensitivity and specificity, contributing to under or over-immunosuppression, respectively, and by the need for invasive biopsies. Herein, we report an approach to reproducibly analyze ileal fluid obtained through stoma sampling for antimicrobial peptide/protein concentrations, reasoning that these molecules may provide an assessment of intestinal homeostasis and levels of intestinal inflammation over time. Concentrations of lysozyme (LYZ), myeloperoxidase (MPO), calprotectin (S100A8/A9) and β-defensin 2 (DEFB2) were assessed using adaptations of commercially available enzyme-linked immunosorbent assays (ELISAs). The concentration of α-defensin 5 (DEFA5) was assessed using a newly developed sandwich ELISA. Our data support that with proper preparation of ileal effluent specimens, precise and replicable determination of antimicrobial peptide/protein concentrations can be achieved for each of these target molecules via ELISA. This approach may prove to be reliable as a clinically useful assessment of intestinal homeostasis over time for patients with ileostomies.

The continuing mutability of the SARS-CoV-2 virus can result in failures of diagnostic assays. To address this, we describe a generalizable bioinformatics-to-biology pipeline developed for the calibration and quality assurance of inactivated SARS-CoV-2 variant panels provided to Radical Acceleration of Diagnostics programs (RADx)-radical program awardees. A heuristic genetic analysis based on variant-defining mutations demonstrated the lowest genetic variance in the Nucleocapsid protein (Np)-C-terminal domain (CTD) across all SARS-CoV-2 variants. We then employed the Shannon entropy method on (Np) sequences collected from the major variants, verifying the CTD with lower entropy (less prone to mutations) than other Np regions. Polyclonal and monoclonal antibodies were raised against this target CTD antigen and used to develop an Enzyme-linked immunoassay (ELISA) test for SARS-CoV-2. Blinded Viral Quality Assurance (VQA) panels comprised of UV-inactivated SARS-CoV-2 variants (XBB.1.5, BF.7, BA.1, B.1.617.2, and WA1) and distractor respiratory viruses (CoV 229E, CoV OC43, RSV A2, RSV B, IAV H1N1, and IBV) were assembled by the RADx-rad Diagnostics core and tested using the ELISA described here. The assay tested positive for all variants with high sensitivity (limit of detection: 1.72-8.78 ng/mL) and negative for the distractor virus panel. Epitope mapping for the monoclonal antibodies identified a 20 amino acid antigenic peptide on the Np-CTD that an in-silico program also predicted for the highest antigenicity. This work provides a template for a bioinformatics pipeline to select genetic regions with a low propensity for mutation (low Shannon entropy) to develop robust 'pan-variant' antigen-based assays for viruses prone to high mutational rates.

[Display omitted] • MOE ameliorates depressive behaviors by modulating the microbiota-gut-brain axis. • MOE restored sucrose preference in depressed mice. • MOE reversed CUMS-induced 5-HT reduction in the prefrontal cortex and hippocampus. • MOE increased gut lactobacilli in stressed mice. The gut microbiota has been posited as a target for the treatment of major depressive disorder. Herein, we investigated the effect of the hydroethanolic leaf extract of Mallotus oppositifolius (MOE) on the gut microbiota of mice and how this contributes to its known antidepressant-like effect. A 6-week chronic unpredictable mild stress (CUMS) procedure was employed in 7 groups of mice to induce depression. From the third week, oral MOE treatments (10, 30, 100 mg/kg) and two reference drugs, fluoxetine (12 mg/kg) and minocycline (40 mg/kg), known to affect the gut microbiota, were administered. The sixth and seventh groups were the vehicle stressed (VEH-S) and non-stressed groups (VEH-NS). Changes in depressive-like behaviors were assessed using sucrose preference test while the forced swimming test (FST) was used to assess sustained antidepressant-effect after treatment discontinuation. Moreover, changes in prefrontal cortex (PFC) and hippocampal serotonin (5-HT) levels were evaluated using enzyme-linked immunosorbent assay (ELISA). The effect of treatment on the profile of the gut microbiota of the groups was elucidated using 16S rRNA Oxford Nanopore sequencing. MOE and reference drugs reversed the depression-associated reduction in sucrose preference when compared to VEH-S. MOE (with peak effect at 30 mg/kg) reduced immobility while increasing swimming and climbing behaviors. MOE reversed CUMS-induced reduction of 5-HT concentration in PFC and hippocampus. The behavioral effects of MOE were associated with shifts in the gut microbiota of CUMS-exposed mice. The study has provided seminal evidence that MOE ameliorates CUMS-induced depressive symptoms by modulating gut microbiota and increasing brain 5-HT levels. [ABSTRACT FROM AUTHOR]

Background: The aim of this study was to investigate the therapeutic effects and mechanisms of PRP-exos combined with cyclic peptide-modified β-TCP scaffold in the treatment of rabbit knee cartilage defect. Methods: PRP-exos were extracted and characterized by TEM, NTA and WB. The therapeutic effects were evaluated by ICRS score, HE staining, Immunohistochemistry, qRT-PCR and ELISA. The repair mechanism of PRP-exos was estimated and predicted by miRNA sequencing analysis and protein–protein interaction network analysis. Results: The results showed that PRP-exos had a reasonable size distribution and exhibited typical exosome morphology. The combination of PRP-exos and cyclic peptide-modified β-TCP scaffold improved ICRS score and the expression level of COL-2, RUNX2, and SOX9. Moreover, this combination therapy reduced the level of MMP-3, TNF-α, IL-1β, and IL-6, while increasing the level of TIMP-1. In PRP-exos miRNA sequencing analysis, the total number of known miRNAs aligned across all samples was 252, and a total of 91 differentially expressed miRNAs were detected. The results of KEGG enrichment analysis and the protein–protein interaction network analysis indicated that the PI3K/AKT signaling pathway could impact the function of chondrocytes by regulating key transcription factors to repair cartilage defect. Conclusion: PRP-exos combined with cyclic peptide-modified β-TCP scaffold effectively promoted cartilage repair and improved chondrocyte function in rabbit knee cartilage defect. Based on the analysis and prediction of PRP-exos miRNAs sequencing, PI3K/AKT signaling pathway may contribute to the therapeutic effect. These findings provide experimental evidence for the application of PRP-exos in the treatment of cartilage defect. [ABSTRACT FROM AUTHOR]

Background The presence of ACPA significantly increases the risk of developing RA. Dysregulation of lymphocyte subpopulations was previously described in RA. Our objective was to propose the predictive model for progression to clinical arthritis based on peripheral lymphocyte subsets and ACPA in individuals who are at risk of RA. Methods Our study included 207 at-risk individuals defined by the presence of arthralgias and either additional ACPA positivity or meeting the EULAR definition for clinically suspect arthralgia. For the construction of predictive models, 153 individuals with symptom duration ≥12 months who have not yet progressed to arthritis were included. The lymphocyte subsets were evaluated using flow cytometry and anti-CCP using ELISA. Results Out of all individuals with arthralgia, 41 progressed to arthritis. A logistic regression model with baseline peripheral blood lymphocyte subpopulations and ACPA as predictors was constructed. The resulting predictive model showed that high anti-CCP IgG, higher percentage of CD4+ T cells, and lower percentage of T and NK cells increased the probability of arthritis development. Moreover, the proposed classification decision tree showed that individuals having both high anti-CCP IgG and low NK cells have the highest risk of developing arthritis. Conclusions We propose a predictive model based on baseline levels of lymphocyte subpopulations and ACPA to identify individuals with arthralgia with the highest risk of progression to clinical arthritis. The final model includes T cells and NK cells, which are involved in the pathogenesis of RA. This preliminary model requires further validation in larger at-risk cohorts. [ABSTRACT FROM AUTHOR]

Objective To examine disease and target engagement biomarkers in the RISE-SSc trial of riociguat in early diffuse cutaneous systemic sclerosis and their potential to predict the response to treatment. Methods Patients were randomized to riociguat (n  = 60) or placebo (n  = 61) for 52 weeks. Skin biopsies and plasma/serum samples were obtained at baseline and week 14. Plasma cyclic guanosine monophosphate (cGMP) was assessed using radio-immunoassay. α-Smooth muscle actin (αSMA) and skin thickness were determined by immunohistochemistry, mRNA markers of fibrosis by qRT-PCR in skin biopsies, and serum CXC motif chemokine ligand 4 (CXCL-4) and soluble platelet endothelial cell adhesion molecule-1 (sPECAM-1) by enzyme-linked immunosorbent assay. Results By week 14, cGMP increased by 94 (78)% with riociguat and 10 (39)% with placebo (P  < 0.001, riociguat vs placebo). Serum sPECAM-1 and CXCL-4 decreased with riociguat vs placebo (P  = 0.004 and P  = 0.008, respectively). There were no differences in skin collagen markers between the two groups. Higher baseline serum sPECAM-1 or the detection of αSMA-positive cells in baseline skin biopsies was associated with a larger reduction of modified Rodnan skin score from baseline at week 52 with riociguat vs placebo (interaction P -values 0.004 and 0.02, respectively). Conclusion Plasma cGMP increased with riociguat, suggesting engagement with the nitric oxide–soluble guanylate cyclase–cGMP pathway. Riociguat was associated with a significant reduction in sPECAM-1 (an angiogenic biomarker) vs placebo. Elevated sPECAM-1 and the presence of αSMA-positive skin cells may help to identify patients who could benefit from riociguat in terms of skin fibrosis. Trial registration Clinicaltrials.gov, NCT02283762. [ABSTRACT FROM AUTHOR]

Objective: Bone incision leads to interrupted and sluggish blood flow in the process of distraction osteogenesis (DO), creating a hypoxia (0–2% oxygen tension) at the center of the bone callus. This hypoxia is critical in the coupling of osteogenesis and angiogenesis during DO. This study aimed to investigate the effect of Uniaxial Static Strain (USS) on osteogenesis in osteoblasts under hypoxic conditions, with a focus on the expression of osteogenic markers and angiogenic factors. Methods: The USS was made by a multi-unit tension compression device.Osteoblasts were subjected to 10% USS made under hypoxic conditions to mimic the process of DO in vitro. The cell proliferation, alkaline phosphatase (ALP) activity, mineralized nodule formation, and expression of osteogenic and angiogenic markers were evaluated by using a CCK-8 assay, alkaline phosphatase (ALP) staining, ALP activity assay, alizarin red S staining, qRT-PCR, Western blotting and ELISA. Results: Hypoxia inhibited osteoblast cell proliferation, ALP activity, mineralized nodule formation, and the expression of runt-related transcription factor 2 (Runx- 2), osteopontin(OPN), osteocalcin (OCN), collagen type I (Col1a1). Conversely, hypoxia upregulated the expression of hypoxia-inducible factor 1-alpha (HIF-1α) and vascular endothelial growth factor (VEGF), which are associated with angiogenesis. However, the application of USS enhanced osteoblasts' osteogenic capacity and upregulated angiogenic factors under hypoxic conditions. Conclusion: USS can enhance osteogenesis in osteoblasts under hypoxic conditions. Moreover, it may stimulate angiogenesis by promoting the expression of VEGF, which further contributes to bone formation. This finding provides important implications for understanding the mechanisms involved in bone regeneration and may have clinical applications in optimizing the effectiveness of DO techniques. [ABSTRACT FROM AUTHOR]

Aim: This study aims to assess the effectiveness of therapy of human synovial membrane-derived MSCs (SM-MSC) from OA grade IV patients in treating knee OA. Methods: SM-MSC were isolated from patients undergoing total knee replacement surgery, cultured to the fourth passage, and characterized using flow cytometry. Differentiation potential was assessed through lineage-specific staining. Osteoarthritis was induced in 24 Wistar rats via monosodium iodoacetate (MIA). The rats were divided into three groups: negative control, OA control, and OA treated with SM-MSC. Radiological, histopathological, and molecular analyses were conducted to evaluate cartilage repair and gene expression. Results: Flow cytometry confirmed the MSC phenotype of SM-MSC, and successful differentiation was observed. Radiological and histopathological analyses showed significant improvement in the SM-MSC treated group, with reduced cartilage damage and higher Safranin O staining compared to the OA control group. Gene expression analysis indicated increased type-2 collagen (COL-2) expression in the SM-MSC treated group, although MMP-13 levels remained unchanged across all groups. Conclusion: Human SM-MSCs from OA grade IV patients significantly improved cartilage repair in an OA rat model, demonstrating their potential as a therapeutic option for OA. To enhance long-term efficacy and anti-inflammatory effects, further studies are needed to optimize treatment protocols, including injection frequency and dosage. [ABSTRACT FROM AUTHOR]

Context.--: Galactose-deficient immunoglobulin A1 (Gd-IgA1) deposition in the renal mesangium plays a role in the pathogenesis of IgA nephropathy. Objective.--: To assess the serum Gd-IgA1 level in biopsy-proven IgA nephropathy cases at diagnosis and 3 months post treatment and its relation with histologic Oxford classification. Design.--: In this hospital-based prospective cohort study, 40 cases and 20 controls were enrolled. Serum samples of biopsy-proven IgA nephropathy cases collected on the day of biopsy and 3 months post treatment were evaluated. Solid-phase ELISA (enzyme-linked immunosorbent assay) was performed for assessment of Gd-IgA1 level. All renal biopsies were scored by using the Oxford classification (C-MEST score). The association of serum Gd-IgA1 levels with other established prognostic parameters was assessed. To estimate the prognostic value of markers, logistic regression analysis and Kruskal-Wallis ANOVA (analysis of variance) were used. Results.--: A significant difference was observed in the serum Gd-IgA1 level values in the IgA nephropathy cases and healthy controls (P = .001) at baseline. However, no significant correlation between serum Gd-IgA1 levels at baseline and 3 months of follow-up (P = .31) or between baseline levels and age, proteinuria, hematuria, or estimated glomerular filtration rate was noted. There was no significant correlation between C-MEST score and serum Gd-IgA1 levels at baseline (P > .05); however, the distribution of Gd-IgA1 at 3 months was found to differ significantly between different grades of S score (P = .008). Conclusions.--: Serum Gd-IgA1 levels may be of utility in predicting disease progression in IgA nephropathy cases. Measurement of serum Gd-IgA1 levels for the diagnosis and prognosis of IgA nephropathy may preclude the need for invasive renal biopsies. [ABSTRACT FROM AUTHOR]

Background: Information about the chikungunya disease burden by age groups and geographic distribution is necessary to guide appropriate control measures. With this, we conducted a systematic review and meta-analysis to estimate the disease burden of chikungunya fever in India. Material and Methods: We conducted this systematic review according to the Cochrane Collaboration guidelines. We retrieved relevant articles from PubMed and a free online search. Two investigators screened titles and abstracts and extracted data from the relevant articles. Our primary outcome is the proportion of laboratory-confirmed Chikungunya fever among clinically suspected patients. We used a random effect model to estimate the pooled proportion of Chikungunya fever. Result: A total of 20 articles were included in the quantitative syntheses. The pooled proportion of laboratory-confirmed chikungunya fever from 20 studies estimated using the random effects model is 24% (95%CI: 15-34%). We found the pooled proportion in the southern region was 35% (95%CI: 4-66%), 28% (95%CI: 3-58%) in the western region, 24% (95%CI: 1-48%) in the eastern region, 20% (95%CI: 12-29%) in the northern region, and 4% (95%CI: 1-6%) in North-eastern region. Conclusion: This review emphasizes the need to strengthen the surveillance of disease burden using multiple diagnostic tests and the need for an appropriate molecular diagnostic for early detection of the chikungunya virus. [ABSTRACT FROM AUTHOR]

Objective: Autoinflammation and phospholipase C (PLC) γ2–associated antibody deficiency and immune dysregulation (APLAID) syndrome is an autoinflammatory disease caused by gain‐of‐function variants in PLCG2. This study investigates the pathogenic mechanism of a novel variant of PLCG2 in a patient with APLAID syndrome. Methods: Whole‐exome sequencing and Sanger sequencing were used to identify the pathogenic variant in the patient. Single‐cell RNA sequencing, immunoblotting, luciferase assay, inositol monophosphate enzyme‐linked immunosorbent assay, calcium flux assay, quantitative PCR, and immunoprecipitation were used to define inflammatory signatures and evaluate the effects of the PLCG2 variant on protein functionality and immune signaling. Results: We identified a novel de novo variant, PLCG2 p.D993Y, in a patient with colitis, pansinusitis, skin rash, edema, recurrent respiratory infections, B‐cell deficiencies, and hypogammaglobulinemia. The single‐cell transcriptome revealed exacerbated inflammatory responses in the patient's peripheral blood mononuclear cells. Expression of the D993Y variant in HEK293T, COS‐7, and PLCG2 knock‐out THP‐1 cell lines showed heightened PLCγ2 phosphorylation; elevated inositol‐1,4,5‐trisphosphate production and intracellular Ca2+ release; and activation of the MAPK, NF‐κB, and NFAT signaling pathways compared with control‐transfected cells. In vitro experiments indicated that the D993Y variant altered amino acid properties, disrupting the interaction between the catalytic and autoinhibitory domains of PLCγ2, resulting in PLCγ2 autoactivation. Conclusion: Our findings demonstrated that the PLCG2 D993Y variant is a gain‐of‐function mutation via impairing its autoinhibition, activating multiple inflammatory signaling pathways, thus leading to APLAID syndrome. This study further broadens the molecular underpinnings and phenotypic spectrum of PLCγ2‐related disorders. [ABSTRACT FROM AUTHOR]

Objective: Autoantibodies are a hallmark of lupus nephritis (LN), but their association with LN classes and treatment response are not adequately known. In this study, we quantified circulating autoantibodies in the Accelerating Medicines Partnership LN longitudinal cohort to identify serological biomarkers of LN histologic classification and treatment response and how these biomarkers change over time based on treatment response. Methods: Peripheral blood samples were collected from 279 patients with systemic lupus erythematosus undergoing diagnostic kidney biopsy based on proteinuria. Of these, 268 were diagnosed with LN. Thirteen autoantibody specificities were measured by bead‐based assays (Bio‐Rad Bioplex 2200) and anti‐C1q by enzyme‐linked immunosorbent assay at the time of biopsy (baseline) and at 3, 6, and 12 months after biopsy. Clinical response was determined at 12 months. Results: Proliferative LN (International Society of Nephrology/Renal Pathology Society class III/IV±V, n = 160) was associated with higher concentrations of anti‐C1q, anti‐chromatin, anti–double‐stranded DNA (dsDNA), and anti–ribosomal P autoantibodies compared to nonproliferative LN (classes I/II/V/VI, n = 108). Anti‐C1q and‐dsDNA were independently associated with proliferative LN. In proliferative LN, higher baseline anti‐C1q levels predicted complete response (area under the curve [AUC] 0.72; P = 0.002) better than baseline proteinuria (AUC 0.59; P = 0.21). Furthermore, all autoantibody levels except for anti‐La/SSB decreased over 12 months in patients with proliferative, but not membranous, LN with a complete response. Conclusion: Baseline levels of anti‐C1q and anti‐dsDNA may serve as noninvasive biomarkers of proliferative LN, and anti‐C1q may predict complete response at the time of kidney biopsy. In addition, tracking autoantibodies over time may provide further insights into treatment response and pathogenic mechanisms in patients with proliferative LN. [ABSTRACT FROM AUTHOR]

Purpose: Androgen receptor (AR) expression and signaling have been regarded as a mechanism for regulating muscle hypertrophy. However, little is known about the associations between acute and chronic changes in skeletal muscle total AR, cytoplasmic AR (cAR), nuclear AR (nAR), and AR DNA-binding (AR-DNA) induced by resistance training (RT) and hypertrophy outcomes in women and men. This study aimed to investigate the acute and chronic effects of RT on skeletal muscle total AR, cAR, and nAR contents and AR-DNA in women and men. In addition, we investigated whether these acute and chronic changes in these markers were associated with muscle hypertrophy in both sexes. Methods: Nineteen women and 19 men underwent 10 wk of RT. Muscle biopsies were performed at baseline, 24 h after the first RT session, and 96 h after the last session. AR, cAR, and nAR were analyzed using Western blotting, and AR-DNA using an ELISA-oligonucleotide assay. Fiber cross-sectional area (fCSA) was analyzed through immunohistochemistry and muscle cross-sectional area (mCSA) by ultrasound. Results: At baseline, men demonstrated greater nAR than women. Baseline cAR was significantly associated with type II fCSA hypertrophy in men. Acutely, both sexes decreased AR and cAR, whereas men demonstrated greater decreases in nAR. After 10 wk of RT, AR, and nAR remained unchanged, men demonstrated greater cAR compared with women, and both sexes decreased AR-DNA activity. Acute and chronic changes in AR markers did not correlate with muscle hypertrophy (type I/II fCSA and mCSA) in women or men. Conclusions: Baseline cAR content may influence hypertrophy in men, whereas neither RT-induced acute nor chronic changes in AR, cAR, nAR, and AR-DNA are associated with muscle hypertrophy in women or men. [ABSTRACT FROM AUTHOR]

Objectives: Osteoarthritis (OA) is the most common chronic joint degenerative disease. Herein, we investigated long non-coding RNA Opa-interacting protein 5-antisense transcript 1's (OIP5-AS1) in regulating mitophagy during OA. Methods: RNA immunoprecipitation and RNA pull-down verified the relationship between molecules. Cell counting kit-8 detected cell viability. Enzyme-linked immunosorbent assay evaluated inflammatory cytokines secretion. Flow cytometry measured the contents of reactive oxygen species (ROS) and calcium. Immunofluorescence staining analysed TOMM20 and LC3B levels. JC-1 staining was adopted to measure mitochondrial membrane potential. The changes of mitophagy were analysed by transmission electron microscopy. Results: Lipopolysaccharide (LPS) treatment contributed to the decrease of chondrocyte viability, and calcium level and inhibited mitochondrial membrane potential, while elevating the secretion of inflammatory factors, ROS, and TOMM20 expression. OIP5-AS1 overexpression inhibited LPS-induced chondrocyte injury and activated mitophagy. OIP5-AS1 upregulated the peroxisome proliferator–activated receptor-γ (PPAR-γ) mRNA level to regulate adenosine monophosphate–activated protein kinase (AMPK)/v-akt murine thymoma viral oncogene homolog (Akt)/mammalian target of rapamycin (mTOR) signalling by interacting with FUS. PPAR-γ overexpression alleviated LPS-induced chondrocyte injury by activating AMPK/Akt/mTOR signalling. PPAR-γ knockdown reversed the promotion of OIP5-AS1 upregulation on mitophagy. Conclusions: OIP5-AS1 promotes PPAR-γ expression to activate the AMPK/Akt/mTOR signalling, thereby enhancing mitophagy and alleviating OA progression. [ABSTRACT FROM AUTHOR]

Introduction: Serum heat shock protein (HSP) concentrations have been reported as potential biomarkers for amyotrophic lateral sclerosis (ALS). Here, we investigate the role of serum HSP70, HSP90, and DNAJC7 as biomarkers for ALS. Methods: Serum samples were collected from ALS patients and volunteer controls from three different clinical cohorts (in Germany, Ireland, and Italy). Serum HSP concentrations were determined using enzyme-linked immunosorbent assay. Descriptive statistics, generalized logistic regression, and Cox proportional hazards models were used to model associations between log serum HSP concentrations and ALS risk. Results: In total, 251 ALS patients and 184 healthy volunteers were included. Logistic regression models failed to find associations between ALS risk and log serum concentration of HSP70 (OR 0.43, 95% CI: 0.10–1.78, p = 0.242), HSP90 (OR 0.95, 95% CI: 0.39–2.37, p = 0.904), or DNAJC7 (OR 1.55, 95% CI: 0.90–2.68, p = 0.118). Survival of ALS patients was not associated with log serum concentration of HSP HSP70 (HR1.06, 95% CI: 0.36–3.14, p = 0.916), HSP90 (HR 1.17, 95% CI: 0.67–2.02, p = 0.584), or DNAJC7 (HR 0.83, 95% CI: 0.57–1.21, p = 0.337). Discussion: We did not replicate previous findings that serum HSP70 and HSP90 concentrations were associated with risk of ALS. DNAJC7 was not associated with ALS risk, and there were no obvious longitudinal patterns in log serum concentrations of HSP70, HSP90, or DNAJC7. In addition, serum HSP concentrations were not associated with ALS survival. [ABSTRACT FROM AUTHOR]

Introduction/objectives: JAK/STAT signaling inhibition exerts therapeutic effects on angiogenesis in rheumatoid arthritis (RA). However, whether the inhibitory effect differs among JAK inhibitors because of differing selectivity is unknown. Therefore, we compared the inhibitory effects of tofacitinib, baricitinib, peficitinib, upadacitinib, and filgotinib on angiogenesis. Method: RA-derived fibroblast-like synoviocytes (RA-FLS) were seeded on type I collagen gel, and human umbilical vein endothelial cells (HUVECs) were directly added. The control and aforementioned JAK inhibitors were added to the medium, followed by stimulation with interleukin (IL)-6 and soluble IL-6 receptor (sIL-6R). Each JAK inhibitor's concentration was determined based on estimated blood concentrations. The vascular endothelial growth factor (VEGF) concentration was evaluated with an enzyme-linked immunosorbent assay using the medium from the first exchange. A migration assay was performed, and HUVEC migration was evaluated using CD31 fluorescence immunostaining. Results: Hematoxylin–eosin staining showed that compared with the non-JAKi treatment group, the JAKi treatment group markedly degenerated in the sub-lining and deep lining, with decreased lymphocyte infiltration and neovascularization [Rooney's score subscale, non-JAKi vs JAKi (median, 6.5 vs 2.5, p = 0.005)]. In vitro, IL-6 and sIL-6R administration increased VEGF production from RA-FLS and promoted neovascularization in HUVECs, and JAK-inhibitor administration, which decreased VEGF production from RA-FLS and suppressed HUVEC migration, inhibited neovascularization in RA-FLS and HUVEC co-cultures. Conclusions: The JAK inhibitors suppressed IL-6-induced angiogenesis via decreased VEGF production and HUVEC migration in RA-FLS and HUVEC co-cultures. No significant differences were observed among the JAK inhibitors, whose anti-angiogenic effect may be an important mechanism for RA treatment. Key Points • JAK inhibitors inhibit angiogenesis in RA by reducing VEGF production from RA-derived fibroblast-like synoviocytes. • Our study provides new insights into RA treatment by elucidating the anti-angiogenic effect of JAK inhibitors. [ABSTRACT FROM AUTHOR]

• Placenta accreta spectrum (PAS), including placenta accreta, increta and percreta, is an important disease group that expresses the pathological adhesion of the placenta to the uterine myometrium and has risks such as maternal bleeding, hysterectomy and death. • There is still no marker to be used in the diagnosis of the disease. • Nuclear factor erythroid 2-related factor 2 (Nrf2)-Kelch-like ECH-related protein 1 (Keap1) pathway has a role in the pathogenesis of many diseases. • The Nrf2-Keap1 pathway is impaired in these patients. Placenta accreta spectrum (PAS) is an important disease group with risks such as maternal bleeding, hysterectomy, and death, which expresses the pathological adhesion of the placenta to the uterine myometrium, including placenta accreta, increta, and percreta, with an increased incidence with an increase in cesarean section rates. In this study, we aimed to investigate the Nuclear factor erythroid 2-related factor 2 (Nrf2)-Kelch-like ECH-related protein 1 (Keap1) pathway in these patients. Serum Sestrin 2, Nrf2, Keap1, glycogen synthase kinase 3β (GSK-3β), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) activities and malondialdehyde-modified low-density lipoprotein (MDA-LDL) levels were performed by the Enzyme-Linked Immunosorbent Assay (ELISA) method. In the findings obtained, Nrf2, Keap1, GSK-3ß, MDA-LDL levels, SOD and GSH-Px activities were statistically significantly different in the patient group compared to the control group. While MDA-LDL values were found to be high in the patient group, Nrf2, Keap1, GSK-3ß levels, SOD and GSH-Px activities were significantly lower, except for Sestrin 2 values. In addition, when grouped according to the degree of invasion, Nrf2 levels were found to be lower and Keap1 levels higher. As a result, it was determined that the Nrf2-Keap1 pathway was disrupted in PAS patients, and the oxidant/antioxidant balance was impaired in the oxidant direction. The results show that Nrf2 and Keap1 parameters can be useful in determining the degree of placental invasion. [ABSTRACT FROM AUTHOR]

The aim of our study was to show the presence of neurotrophic factors in breast milk that have a significant impact on neurocognitive development of children aged two years and beyond. Mothers expressed at least 5 mL of breast milk into sterile containers when their children 18, 24, and ≥ 25 months of age, and then specimens were transferred to Eppendorf tubes and stored at -20 °C. One day before the analysis, specimens were kept at +4 °C and then thawed at room temperature to prepare them for analysis. Brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF), and S100B neurotrophic growth factor levels were analyzed using the sandwich enzyme-linked immunosorbent assay (ELISA) principles. Sixty-two mothers with children aged 18 months were included in the study. The mean age of the mothers was 33.4 (± 0.71) years. Due to the detection limits of the commercial kits, BDNF and S100B analyses could not be conducted. Therefore, only GDNF was analyzed. The presence of GDNF was found in the breast milk samples taken at 18, 24, and ≥ 25 months, and the median (min max) values were 315,505 ng/mL (193,067 750,718), 316,721 ng/mL (161,278 l-752,252), and 564,577 ng/mL (238,528-781,104) respectively. There were no significant differences between GDNF levels of breast milk samples collected from the same mother at the three different time points (18, 24, and ≥ 25 months) (p = 0.278). Conclusion: Our study was the first to show the presence of neurotrophic factors in the breast milk of mothers with healthy children over one year of age. Our results provide evidence-based data on the importance of breastfeeding until children are at least two years of age. What is Known: • Presence of Brain-derived neurotrophic factor (BDNF), glial cell line-derived neurotrophic factor (GDNF), and S100B neurotrophic growth factor have been shown in the breast milk of mothers whose infants are the first year of life. What is New: • Glial Cell Line-Derived Neurotrophic factors continue to present in breast milk of mothers with children aged 18, 24, and ≥ 25 months, without any significant difference in level between months. [ABSTRACT FROM AUTHOR]

Trastuzumab is a potent targeted therapy drug for HER2-positive cancer patients. A comprehensive understanding of trastuzumab's mechanism of action, pharmacokinetic (PK) parameters, and steady-state exposure in different treatment regimens and administration routes is essential for a thorough evaluation of the drug's safety and effectiveness. Due to the distinctive pharmacokinetics, indications, and administration methods of trastuzumab, this understanding becomes crucial. Drug exposure can be assessed by measuring trastuzumab's peak concentration, trough concentration, or area under the curve through assays like enzyme-linked immunosorbent assay (ELISA) or liquid chromatography-tandem mass spectrometry (LC-MS/MS). The dose-response (D-R) and exposure-response (E-R) relationships establish the correlation between drug dosage/exposure and the therapeutic effect and safety. Additionally, various covariates such as body weight, aspartate transaminase, and albumin levels can influence drug exposure. This review provides a comprehensive overview of trastuzumab's mechanism of action, data on steady-state concentration and PK parameters under multiple administration routes and indications, discussions on factors influencing PK parameters, and evaluations of the effectiveness and safety of E-R and D-R in diverse HER2-positive cancer patients. [ABSTRACT FROM AUTHOR]

Due to the ban of certain single-use plastics in the European Union, food contact materials (FCM) from biobased and/or biodegradable polymers are increasingly being used. Some FCM are made from wheat or rye and therefore contain gluten, which is a food allergen and known to cause celiac disease. Legislation currently does not require allergen labelling on FCM and there is only some first information that gluten from FCM can migrate into gluten-free foods. Our aim was to analyze the extent of gluten migration from six different FCM into a variety of liquid and solid foods to assess the risk of exposure for wheat allergy and celiac disease patients. We show that the extent of gluten migration depended on the properties of the material, the liquid or solid food it comes into contact with and contact time. There was no clear effect of temperature or pH value. Of the six FCM studied, wheat bran-based plates had the highest potential to release gluten with concentrations of up to 203.0 mg/L of gluten in deionized water after 30 min. To protect patients, it is important to raise awareness of the risk of gluten migration from such FCM and help patients identify and avoid gluten-containing FCM. Further, legislation needs to be adapted urgently to include mandatory labelling of allergens on these biodegradable FCM. [ABSTRACT FROM AUTHOR]

Aims: Hepatitis E caused by the hepatitis E virus (HEV) is prevalent worldwide. In China, considerable shifts in the epidemiology of hepatitis E have been observed over the last two decades, with ongoing changes in the prevalence of HEV. Method s : This study, in conjunction with the health examinations for students and workers, aims to estimate the seroprevalence and assess the risk factors of HEV infection in general population in Hebei province, China. Epidemiological information was collected using a specific questionnaire and blood samples were collected from each participant during the process of health examination. Anti‐HEV IgG and IgM in sera were tested using the Wantai ELISA assay kits. Logistic regression modelling was used to identify associated risk factors. Results: The average positive rate of anti‐HEV IgG in students (6–25 years) was 3.4%. One (0.2%) student was anti‐HEV IgM positive, while also testing positive for IgG. The HEV seroprevalence was not related to students' gender, school, or family residence. In occupational populations, the overall seropositivity rate was 13.3% for anti‐HEV IgG and 0.67% for IgM. HEV seropositivity increased significantly with age, ranging from 3.8% to 18.6% in age groups, and differed significantly among four occupation groups: farmers (17.6%), food supply workers (18.0%), other non‐farm workers (14.7%) and healthcare workers (5.9%) (p = 0.002). Multivariable logistic analysis confirmed the significant correlations of seroprevalence with age and occupation. Conclusions: The study found a low seroprevalence of HEV in children and young adults in Hebei Province, China. Advanced age correlates with higher seroprevalence in occupational populations, indicating an accumulation of HEV infection over time. Seroprevalence varied significantly among different occupation groups, suggesting the important role of occupational exposure for HEV infection. [ABSTRACT FROM AUTHOR]

Atopic dermatitis is a common allergic skin disease characterized by eczema, dryness, and severe itching, which has a significant impact on patients' quality of life. Its pathogenesis involves immune dysregulation and impaired skin barrier function. In this study, the therapeutic potential of Ampelopsis brevipedunculata (Maxim.) Trautv for alleviating atopic dermatitis symptoms was investigated using the NC/Nga mouse model. A. brevipedunculata (Maxim.) Trautv was effective in attenuating the atopic dermatitis-like epidermal thickening and the immune cell infiltration in the skin lesions induced by the atopic dermatitis ointment. Additionally, A. brevipedunculata (Maxim.) Trautv suppressed the production of inflammatory mediators and chemokines associated with atopic dermatitis progression. In addition, A. brevipedunculata (Maxim.) Trautv exhibited restorative effects on skin barrier dysfunction by increasing the expression of key barrier proteins. Mechanistically, A. brevipedunculata (Maxim.) Trautv modulated the mitogen-activated protein kinase pathway and inhibited nuclear factor-kappa B activation. These findings highlight the potential of A. brevipedunculata (Maxim.) Trautv as a promising therapeutic agent for treating atopic dermatitis and provide insight into its mechanisms of action. [ABSTRACT FROM AUTHOR]

Sepsis has a high mortality rate and leads to multi-organ failure, including lung injury. Inactive rhomboid protease family protein (iRhom2) has been identified as accountable for the release of TNF-α, a crucial mediator in the development of sepsis. This study aimed to evaluate the role of iRhom2 in sepsis and sepsis-induced acute lung injury (ALI). TNF-α and IL-6 secretion in vitro by peritoneal macrophages from wild-type (WT) and iRhom2 knoukout (KO) mice was assessed by enzyme-linked immunosorbent assay. Cecal ligation and puncture (CLP)-induced murine sepsis model was used for in vivo experiments. To evaluate the role of iRhom2 deficiency on survival during sepsis, both WT and iRhom2 KO mice were monitored for 8 consecutive days following the CLP. For histologic and biochemical examination, the mice were killed 18 h after CLP. iRhom2 deficiency improved the survival of mice after CLP. iRhom2 deficiency decreased CD68+ macrophage infiltration in lung tissues. Multiplex immunohistochemistry revealed that the proportion of Ki-67+ CD68+ macrophages was significantly lower in iRhom2 KO mice than that in WT mice after CLP. Moreover, CLP-induced release of TNF-α and IL-6 in the serum were significantly inhibited by iRhom2 deficiency. iRhom2 deficiency reduced NF-kB p65 and IκBα phosphorylation after CLP. iRhom2 deficiency reduces sepsis-related mortality associated with attenuated macrophage infiltration and proliferation in early lung injury. iRhom2 may play a pivotal role in the pathogenesis of sepsis and early stage of sepsis-induced ALI. Thus, iRhom2 may be a potential therapeutic target for the management of sepsis and sepsis-induced ALI. [ABSTRACT FROM AUTHOR]

Early diagnosis of pregnancy is directly related to cost-effective livestock production. We produced a rat monoclonal antibody (mAb) against synthesized porcine early pregnancy factor (pEPF) using conventional hybridoma technology and used it as a tool for the detection of early pregnancy in Duroc sows. The rat pEPF-mAb showed reactivity to uterine tissues of pregnant sows 20 or 30 days post-mating (day 0 defined as the day of mating) and non-pregnant sows (confirmed signs of estrus) in western blotting. Immunohistochemical analysis confirmed that pEPF was located in the stromal and grand epithelial tissues of pregnant sows 20 or 30 days post-mating. In the enzyme-linked immunosorbent assay, pEPF expression in urine and blood showed similar results, with the highest expression observed in pregnant sows 20 days post-mating, whereas there was no significant difference in expression levels between non-pregnant sows and pregnant sows 30 days post-mating. The pEPF-mAb-based pregnancy diagnostic kit can be applied to pig urine samples non-invasively collected at 20 days post-mating with 70 % accuracy. Further improvements to the kit's diagnostic performance may lead to substantial benefits for the swine industry, facilitating more efficient and accurate reproductive management. • Monoclonal antibody for porcine early pregnancy factor (pEPF) was produced with hybridoma technology. • pEPF was detected in uterine tissue of pregnant sows on day 20 and found to be localized in the uterine stromal tissues. • PEPF antibody-based immunochromatographic kit was designed to diagnose Day-20 sow pregnancy using urine samples. [ABSTRACT FROM AUTHOR]

Background: The anti-PD-L1 antibody durvalumab has been approved for use in first-line advanced biliary duct cancer (ABC). So far, predictive biomarkers of efficacy are lacking. Methods: ABC patients who underwent gemcitabine-based chemotherapy with or without durvalumab were retrospectively enrolled, and their baseline clinical pathological indices were retrieved from medical records. Overall (OS) and progression free survival (PFS) were calculated and analyzed. The levels of peripheral biomarkers from 48 patients were detected with assay kits including enzyme-linked immunosorbent assay. Genomic alterations in 27 patients whose tumor tissues were available were depicted via targeted next-generation sequencing. Results: A total of 186 ABC patients met the inclusion criteria between January 2020 and December 2022 were finally enrolled in this study. Of these, 93 patients received chemotherapy with durvalumab and the rest received chemotherapy alone. Durvalumab plus chemotherapy demonstrated significant improvements in PFS (6.77 vs. 4.99 months; hazard ratio 0.65 [95% CI 0.48–0.88]; P = 0.005), but not OS (14.29 vs. 13.24 months; hazard ratio 0.91 [95% CI 0.62–1.32]; P = 0.608) vs. chemotherapy alone in previously untreated ABC patients. The objective response rate (ORR) in patients receiving chemotherapy with and without durvalumab was 19.1% and 7.8%, respectively. Pretreatment sPD-L1, CSF1R and OPG were identified as significant prognosis predictors in patients receiving durvalumab. ADGRB3 and RNF43 mutations were enriched in patients who responded to chemotherapy plus durvalumab and correlated with superior survival. Conclusion: This retrospective real-world study confirmed the clinical benefit of durvalumab plus chemotherapy in treatment-naïve ABC patients. Peripheral sPD-L1 and CSF1R are promising prognostic biomarkers for this therapeutic strategy. Presence of ADGRB3 or RNF43 mutations could improve the stratification of immunotherapy outcomes, but further studies are warranted to explore the underlying mechanisms. [ABSTRACT FROM AUTHOR]

Objective: To investigate the inhibitory effect of Celastrus orbiculatus extracts (COE) on the proliferation of lymphoma cells and the immune regulation ability on inflammation and thrombophilia in vivo. Methods: The 38B9 lymphoma cells were treated with COE (160 µ g/mL) and CTX (25 µ mol/L). The apoptosis rate and cell cycle of each group were detected by flow cytometry. The secretion of inflammatory factors, including interleukin (IL)-6, IL-10, and tumor necrosis factor α (TNF-α), in cell supernatant was detected by enzyme-linked immunosorbent assay (ELISA). In vivo, BALB/c mice were subcutaneously injected with 38B9 lymphoma cells to establish lymphoma model. COE (3 mg·kg−1·d−1) and CTX (40 mg·kg−1·d−1) were administered to the model mice, respectively. The expression of plasma inflammatory factors (IL-6, IL-10 and TNF-α) and thrombus indexes, including D-dimer (D-D), von Willebrand factor (vWF) and tissue factor (TF), were detected by ELISA before tumor bearing (1 d), after tumor formation (14 d) and after intervention (21 d). PicoGreen dsDNA was used to detect the level of serum neutrophil extracellular traps (NETs). Flow cytometry was used to detect the expression of platelet activation marker calcium-dependent lectin-like receptor 2 (CLEC-2). The tumor growth and survival of mice were recorded. Results: The 38B9 lymphoma cells were apoptotic after the intervention of COE and CTX. The ratio of G2-M phase cells decreased in COE intervented cells compared with the control cells (P<0.05), and S phase cells decreased in CTX intervented cells (P<0.05). Also, the secretion level of IL-6 was significantly reduced after COE or CTX intervention (P<0.05), and IL-10 was significantly increased (P<0.05). Furthermore, the tumor mass was reduced, and the median survival time was longer in COE and CTX intervented tumor-bearing mice than in non-intervented mice. The significantly lower levels of TNF-α, IL-6, NETs, TF, DD and CLEC-2, as well as higher IL-10 were observed in COE and CTX treatment mice in comparision with the control mice (P<0.05). Conclusion: COE has a mild and stable anti-tumor effect, which can reduce the secretion of inflammatory factors by lymphoma cells and regulate thrombophilic state caused by tumor inflammatory microenvironment. [ABSTRACT FROM AUTHOR]

Objective: To observe the protective effect and mechanism of hydroxyl safflower yellow A (HSYA) from myocardial ischemia-reperfusion injury on human umbilical vein endothelial cells (HUVECs). Methods: HUVECs were treated with oxygen-glucose deprivation reperfusion (OGD/R) to simulate the ischemia reperfusion model, and cell counting kit-8 was used to detect the protective effect of different concentrations (1.25–160 µ mol/L) of HSYA on HUVECs after OGD/R. HSYA 80 µ mol/L was used for follow-up experiments. The contents of inflammatory cytokines interleukin (IL)-18, IL-1 β, monocyte chemotactic protein 1 (MCP-1), tumor necrosis factor α (TNF-α) and IL-6 before and after administration were measured by enzyme-linked immunosorbent assay. The protein expressions of toll-like receptor, NOD-like receptor containing pyrin domain 3 (NLRP3), gasdermin D (GSDMD) and GSDMD-N-terminal domain (GSDMD-N) before and after administration were detected by Western blot. NLRP3 inflammasome inhibitor cytokine release inhibitory drug 3 sodium salt (CRID3 sodium salt, also known as MCC950) and agonist were added, and the changes of NLRP3, cysteine-aspartic acid protease 1 (Caspase-1), GSDMD and GSDMD-N protein expressions were detected by Western blot. Results: HSYA inhibited OGD/R-induced inflammation and significantly decreased the contents of inflammatory cytokines IL-18, IL-1 β, MCP-1, TNF-α and IL-6 (P<0.01 or P<0.05). At the same time, by inhibiting NLRP3/Caspase-1/GSDMD pathway, HSYA can reduce the occurrence of pyroptosis after OGD/R and reduce the expression of NLRP3, Caspase-1, GSDMD and GSDMD-N proteins (P<0.01). Conclusions: The protective effect of HSYA on HUVECs after OGD/R is related to down-regulating the expression of NLRP3 inflammasome and inhibiting pyroptosis. [ABSTRACT FROM AUTHOR]

Background & Objective: In 2020, breast cancer claimed the lives of approximately 684,996 women worldwide. The use of cancer biomarkers has greatly improved the effectiveness of treatment and early detection. The present study was conducted aimed to identify the role of testin as a potential biomarker in women with breast cancer. Materials & Methods: In this cross-sectional study, 60 women with breast cancer participated by providing serum samples for measuring testin concentrations. Additionally, a control group of 20 healthy individuals was selected. The testin protein concentration was detected using the serum ELISA method. Results: A significant difference was observed between the age groups of patients and healthy women. The majority of women with breast cancer were in their thirties to sixties. On the other hand, there were fewer breast cancer cases recorded in the twenties, with only 1 (1.7%) case out of the total study population of 60 (100%). This difference was found to be statistically significant (P=0.01). The study found that there was no significant difference in the concentration of testin among women with breast cancer who were newly diagnosed compared to those who had received different numbers of treatment doses (1, 3, 5, 7 doses). Also, the results indicated a significant increase in testin concentration among women with breast cancer compare with healthy women. Conclusion: The present study discovered that breast cancer patients have higher levels of testin compared to the healthy women. There is a growing body of evidence suggesting that testin contributes to the development of breast cancer, making it an appealing protein to focus on for cancer treatment. [ABSTRACT FROM AUTHOR]

Background & Objective: Endometrial cancer (EC) is the most prevalent gynecological malignancy in more developed countries. Multiple researches have been done about the role of IL-27 in different cancers that suggest a dual role for this cytokine. In the present study, we evaluated the serum level of IL-27 in endometrial cancer patients. We also investigated the correlations between serum levels of IL-27 and the demographic and clinicopathologic features of the patients. Materials & Methods: In this case-control study, 65 endometrial cancer patients and 58 sex-age-match healthy controls were investigated. Serum levels of IL-27 in both cases and the control group were assessed by a reliable and specific sandwich enzymelinked immunosorbent assay (ELISA) kit and results were analyzed with SPSS. Results: We observed that the serum level of IL-27 in EC patients was dramatically higher than in the control group (P=0.003). Additionally, Higher grades of EC (grade II and III) showed higher IL-27 serum levels compared to the control (P=0.006 and P=0.01 respectively). No significant correlations between serum levels of IL-27 and lymph node involvement, tumor stage, tumor size, and demographic features of the patients were detected. Conclusion: Our results showed that there is a statistically significant difference between serum levels of IL-27 in EC patients and controls. Therefore, the serum level of IL-27 may exert a role in the pathogenesis of endometrial carcinoma, although further studies are needed. [ABSTRACT FROM AUTHOR]

Background: Previous studies have revealed the importance of CYFRA 21–1 in the diagnosis of oral squamous cell cancer (OSCC). However, the results are inconsistent. This meta-analysis is to evaluate CYFRA 21–1's efficacy in distinguishing OSCC. Methods: Systematic searches of Web of Science, PubMed, and CNKI (1996–2024) were conducted following the Preferred Reporting ltems for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. Results: Analysis of 693 patients and 548 controls yielded combined sensitivity (SEN) of 0.71 (95% CI: 0.68, 0.75), specificity (SPE) of 0.88 (95% CI: 0.85, 0.90), and area under the curve (AUC) of 0.927. Subgroup analysis showed higher SEN (0.88), SPE (0.93), and AUC (0.962) in saliva versus serum. Enzyme-linked immunosorbent assay (ELISA) demonstrated superior performance over electrochemiluminescent immunoassay (ECLIA) (AUC: 0.968 vs. 0.868). Conclusion: CYFRA 21–1 is effective in OSCC diagnosis, with ELISA showing better sensitivity. Saliva emerges as a promising diagnostic medium compared to serum. Registration: PROSPERO CRD42024566835 [ABSTRACT FROM AUTHOR]

Osteoarthritis (OA), degenerative joint disease, is the most prevalent form of arthritis worldwide. Besides its substantial burden on society, the high OA morbidity greatly diminishes patients' quality of life. According to recent research, patients-derived serum extracellular vesicles (EVs) are critically involved in sustaining the corresponding disease progression. However, limited research has fully explored the specific functions and molecular mechanisms of OA serum-derived EVs in disease progression. Consequently, we aimed to investigate the underlying mechanism of OA rats-derived serum EVs in regulating OA progression. Before constructing the exosome-cell co-culture system, EVs were extracted from OA and control rat serum and co-cultured with bone marrow mesenchymal stem cells (BM-MSCs). Western blotting (WB), RT-qPCR, and enzyme-linked immunosorbent assay (ELISA) results revealed that OA rat serum-derived EVs upregulated cell pyroptosis-related markers, including nod-Like receptor protein-3 (NLRP3), apoptosis-associated speck-like protein (ASC), gasdermin D (GSDMD), and cleaved caspase-1. The OA rat-EVs also induced the release of LDH and inflammatory cytokines, including interleukin (IL)-1β, IL-18, IL-6, and TNF-α. Additional experiments revealed that OA rat-EVs delivered miR-133a-3p to BM-MSCs and upregulated miR-133a-3p to degrade sirtuin 1 (SIRT1), and activating the downstream NF-κB signaling pathway. Furthermore, the rescuing experiments confirmed that silencing SIRT1 abrogated the miR-133a-3p-induced protective effects in OA-EVs-treated BM-MSCs. In conclusion, OA rats-derived miR-133a-3p-containing EVs modulated the downstream SIRT1/NF-κB pathway-mediated pyroptotic cell death and inflammation in OA. In other words, this study confirmed the role and underlying mechanisms by which OA-associated serum EVs regulate pyroptosis and inflammation response in OA development. [ABSTRACT FROM AUTHOR]

Introduction: Alzheimer’s disease (AD) is the leading cause of dementia, and currently, no effective treatments are available to reverse or halt its progression in clinical practice. Although a plethora of studies have highlighted the benefits of physical exercise in combating AD, elder individuals often have limited exercise capacity. Therefore, mild physical exercise and nutritional interventions represent potential strategies for preventing and mitigating neurodegenerative diseases. Our research, along with other studies, have demonstrated that platycodin D (PD) or its metabolite, platycodigenin, derived from the medicinal plant Platycodon grandiflorus, exerts neuroprotective effects against amyloid β (Aβ)-induced neuroinflammation. However, the combined effects of PD and physical exercise on alleviating AD have yet to be explored. The current study aimed to investigate whether combined therapy could synergistically ameliorate memory deficits and AD pathology in 5 × FAD mice. Methods: Five-month-old 5 × FAD mice were randomly assigned to four groups, and received either PD (5 mg/kg/day, p.o.), voluntary running, or a combination of both for 47 days. Nest building test, locomotion test, and Morris water maze test were used to evaluate the cognitive function. Immunohistochemical and ELISA analysis was performed to determine Aβ build-up, microglia and astrocytes hyperactivation, and survival neurons in the hippocampus and perirhinal cortex. Real-time quantitative PCR analysis was used to assess the polarization of microglia and astrocytes. HPLC analysis was performed to measure monoamine neurotransmitters in the hippocampus. Results and discussion: The combination of PD and voluntary running synergistically restored nest-building behavior, alleviated recognition and spatial memory deficits, and showed superior effects compared to monotherapy. In addition, the PD and voluntary running combination reduced Aβ build-up, decreased hyperactivation of microglia and astrocytes in the hippocampus and perirhinal cortex, promoted the polarization of inflammatory M1 microglia and reactive astrocytes toward beneficial phenotypes, and lowered systemic circulating pro-inflammatory cytokines while increasing anti-inflammatory cytokines in 5 × FAD mice. Furthermore, combined therapy effectively protected neurons and increased levels of 5-hydroxytryptamine (5-HT) and dopamine (DA) in the hippocampus of 5 × FAD mice. In conclusion, the combination of PD and voluntary running holds great potential as a treatment for AD, offering promise for delaying onset or progression of AD. [ABSTRACT FROM AUTHOR]

Background: Genetic factors, including the Hedgehog Interacting Protein (HHIP) gene, play a crucial role in Chronic Obstructive Pulmonary Disease (COPD) susceptibility. This study examines the association between HHIP gene polymorphisms and COPD susceptibility, phenotypes, and serum IL-17 and IL-18 levels in a Han Chinese population. Methods: A case-control study was conducted with 300 COPD patients and 300 healthy controls in Chinese Han population. Participants underwent genotyping for HHIP gene polymorphisms, pulmonary function tests, and quantitative CT scans. DNA samples were sequenced using a custom chip targeting the HHIP gene. Serum IL-17 and IL-18 levels were measured by enzyme-linked immunosorbent assay. Associations between SNPs, COPD susceptibility, and phenotypes were analyzed using logistic and multiple linear regression models, adjusting for confounders. Results: Our study identified the rs11100865 polymorphism in the HHIP gene as significantly associated with COPD susceptibility (OR 2.479, 95% CI 1.527–4.024, P = 2.39E-04) after screening 114 SNPs through rigorous quality control. Stratified analyses further indicated this association was particularly in individuals aged 60 or older. Serum levels of IL-17 and IL-18 were significantly elevated in COPD patients compared to controls, with rs11100865 showing a notable association with IL-18 levels (B = 49.654, SE = 19.627, P = 0.012). However, no significant associations were observed between rs11100865 and serum IL-17 levels, COPD-related imaging parameters, or clinical phenotypes. Conclusion: This study identified a significant association between HHIP gene polymorphisms and COPD susceptibility in a Han Chinese population, with connections to inflammation, but found no significant associations between this SNP and COPD-related imaging or clinical phenotypes. Trial registration: www.chictr.org.cn ID: ChiCTR2300071579 2023-05-18. [ABSTRACT FROM AUTHOR]

Background: Metabolic syndrome (MetS) is one of the most significant public health issues worldwide, which increases the risk of various diseases. Epidemiological evidence suggests that newer anthropometric measures, such as a body shape index (ABSI) and body roundness index (BRI) can be used to predict MetS. However, anthropometric measures to predict the risk for MetS should be clarified in each population. Therefore, this study aimed to examine the association between ABSI, BRI, and MetS risk factors among overweight and obese Iranian adults. Methods: This cross-sectional study included 347 overweight and obese individuals [body mass index (BMI) > 25 kg/m2] aged 20–50 years in Tabriz, Iran. Anthropometric measures were assessed, including BMI, waist circumference (WC), and waist-to-hip ratio (WHR). Additionally, ABSI and BRI were calculated based on the collected data. Blood pressure was measured using standard protocols. Body composition also was measured using body impedance analysis (BIA). Enzymatic-colorimetric methods were used to assess serum glucose and lipids, and enzyme-linked immunosorbent assay (ELISA) kits were used to measure insulin levels. Results: Participants with higher ABSI exhibited significantly higher systolic blood pressure (SBP) (P = 0.001), diastolic blood pressure (DBP) (P = 0.010), and triglyceride (TG) levels (P < 0.001), along with significantly lower high-density lipoprotein cholesterol (HDL-C) levels (P < 0.001). In the crude model, individuals in the highest ABSI tertile (tertile 3) had a higher likelihood of having higher SBP (OR: 1.032; 95% CI: 1.014–1.051) and DBP (OR: 1.33; 95% CI: 1.009–1.058), as well as a significant association with lower HDL-C levels (OR: 0.945; 95% CI: 0.918–0.973). Additionally, both ABSI tertile 2 (OR: 1.005; 95% CI: 1.001–1.008) and tertile 3 (OR: 0.993; 95% CI: 1.003–1.011) were linked to a higher likelihood of having higher TG levels. No significant associations were found between BRI tertiles and MetS risk factors. Conclusion: According to our results, ABSI and BRI are poor predictors of MetS risk variables, in overweight and obese individuals. High ABSI is only slightly linked with high SBP, DBP, and TG and low HDL-C. However, longitudinal and long-term investigations are encouraged to verify the efficacy of these two measures. [ABSTRACT FROM AUTHOR]

Background and Objectives Materials and Methods Results Conclusion The World Health Organization (WHO) African Region accounts for 94% of malaria cases globally, with variability recognized within endemic regions. To determine the detection rate of Plasmodium RNA in blood donors resident in malaria‐endemic areas, samples from three African countries were tested using a Plasmodium nucleic acid test.Whole blood (WB) samples collected from routine donors in Cameroon, Madagascar and Mali were shipped frozen to the United States. Samples were tested individually from WB lysates with the Procleix Plasmodium assay (transcription‐mediated amplification [TMA]). Reactive samples were considered either repeat reactive or initial reactive only, depending on TMA‐retest results. When available, matching plasma samples were tested for Plasmodium antibodies by enzyme immunoassay (EIA).Plasmodium repeat reactivity ranged from 41% (91/223 tested) in Cameroon to 12% (26/216) in Mali and 1% (3/249) in Madagascar. Initially reactive samples, where reactivity did not repeat, were identified from Cameroon (5/223; 2%) and Mali (2/216; 1%). The matched‐plasma subgroup had EIA reactivity ranging from 86% (113/131 tested) in Cameroon to 59% (10/17) in Mali and 27% (68/248) in Madagascar.Plasmodium ribosomal RNA (rRNA) detection and antibody rates varied greatly in the three countries studied. Detection of Plasmodium rRNA can provide an additional tool to address malaria risk in blood donors. [ABSTRACT FROM AUTHOR]

Staphylococcal food poisoning is a significant foodborne illness caused by staphylococcal enterotoxins (SEs). Immunoassays have become the primary method for rapidly detecting harmful bacteria and toxins because of their excellent sensitivity and specificity. However, these assays have limitations in that they cannot differentiate between types of SEs and do not provide rapid, on-site, quantitative testing. In this study, a time-resolved fluorescence immunochromatography assay (TRFICA) was developed specifically for detecting staphylococcal enterotoxin E (SEE), which is commonly found in dairy products. Compared with a double antibody sandwich enzyme-linked immunosorbent assay, which had a detection limit of 0.028 ng/mL, TRFICA demonstrated comparable sensitivity, enabling SEE quantification with a detection limit as low as 0.081 ng/mL in infant formula. Validation by spiking infant formula samples confirmed no cross-reactivity with analogs (recoveries ranged from 93.17% to 128.77%). Furthermore, with an 8-min reaction time and interpretation delivered by a portable TRFICA strip reader, our method demonstrates potential for use in mobile and on-site detection. This study describes a rapid, easy, and reliable method for detecting trace levels of SEE in infant formula, which could serve as an early screening tool toward preventing food poisoning in infants and children. [ABSTRACT FROM AUTHOR]

AbstractHuman Papillomavirus (HPV) is the principal cause of cervical cancer, which is still a prime universal health concern, especially in underdeveloped nations where access to advanced screening techniques is often restricted. Herein, we propose an electrochemical immunosensor for the determination of HPV 16 L1 protein, a key biomarker associated with cervical carcinogenesis. The developed immunosensor, employing glassy carbon electrode (GCE) enhanced with gold nanoparticles, provides a highly sensitive, specific, and cost-effective approach for early screening of cervical cancer. An optimized stepwise fabrication process which includes electrode surface modification and antibody immobilization, was used to enhance sensor performance. Characterization studies confirmed the effective modification of electrodes and the selectivity of immunoreactions. With an impressive limit of detection (LOD) of 0.033 fg mL−1, the immunosensor showed a linear response to L1 protein concentrations ranging from 0.1 fg mL−1 to 1 ng mL−1. Additionally, the immunosensor exhibits excellent specificity, stability, reproducibility, and regeneration capabilities. The performance of the immunosensor was compared with the enzyme-linked immunosorbent assay (ELISA) by analyzing clinical samples (positive: n = 12; healthy: n = 5) showed satisfactory results. The findings indicated that the proposed sensing platform has the potential to be utilized for the early diagnosis of cervical cancer. [ABSTRACT FROM AUTHOR]

Hepatitis C virus (HCV) transmission remains a significant public health concern. It is well documented globally, however, Nowshera district, Pakistan, is lacking such profile. This study aims to explore the relationship between HCV infection and several risk factors, including socio-demographic, clinical and personal life-style factors. This study using a cross-sectional design, examined 606 randomly selected individuals visiting the Pathology department at Qazi Hussain Medical Complex and District Headquarter (DHQ) Hospital Nowshera between May 1, 2022 and Jun 30, 2023. This research investigation employed a methodical approach involving formal interviews in conjunction with structured questionnaires to gather comprehensive information related to socio-demographic characteristics, clinical history, and personal hygiene practices. We collected 10 ml of blood samples and tested the separated serum to identify markers using the Immuno-Chromatographic Test (ICT) and the Enzyme-Linked Immunosorbent Assay (ELISA). IBM SPSS Statistics version 27 was used for data analysis. To measure the association between dependent variable and independent variables, a chi-square and risk analysis was carried out; multiple logistic regression was employed for modeling the risk factors associated with independent variable. A statistical significance level was established at a P-value of less than 0.05. In present study, which included 606 participants, 12% were found to be infected with HCV. Importantly it revealed that facial shave at barber (adjusted odds ratio, aOR = 40.65, p = 0.000) represents the most prevalent mode of HCV transmission. Additionally, a strong association was observed between HCV infection and history of past surgery (RR = 2.98, p = 0.001), HCV infected family member (aOR = 4.28, p = 0.001), workplace injuries (aOR = 6.68, p = 0.000), history of hospital admission (RR = 2.09), practicing ear and nose piercing (aOR = 5.01, p = 0.001), dental treatment (RR = 2.31) and the frequency of injections (aOR = 8.607, p = 0.000). These findings underscore the pressing need for targeted interventions. This study highlights the paramount importance of understanding the modes of HCV transmission and their associated risk factors. The results emphasize the need for educational initiatives, both within the healthcare sector and among the general public, to combat HCV transmission effectively. By implementing sterilization procedures and increasing awareness, we can make significant strides in reducing the burden of HCV infection. Moreover, proactive measures within families can help contain the spread of the virus, ultimately contributing to improved public health. [ABSTRACT FROM AUTHOR]

Background: In addition to its critical role in neurogenesis, brain-derived neurotrophic factor (BDNF) modulates pain and depressive behaviors. Methods: In a translational perspective, we tested the anti-migraine activity of highly purified and characterized recombinant human BDNF (rhBDNF) in an animal model of cephalic pain based on the chronic and intermittent NTG administration (five total injections over nine days), used to mimic recurrence of attacks over a given period. To achieve this, we assessed the effects of two doses of rhBDNF (40 and 80 µg/kg) administered intranasally to adult male Sprague–Dawley rats, on trigeminal hyperalgesia (by orofacial formalin test), gene expression (by rt-PCR) of neuropeptides and inflammatory cytokines in specific areas of the brain related to migraine pain. Serum levels of CGRP, PACAP, and VIP (by ELISA) were also evaluated. The effects of rhBDNF were compared with those of sumatriptan (5 mg/kg i.p), administered 1 h before the last NTG administration. Results: Both doses of rhBDNF significantly reduced NTG-induced nocifensive behavior in Phase II of the orofacial formalin test. The anti-hyperalgesic effect of intranasal high-dose rhBDNF administration in the NTG-treated animals was associated with a significant modulation of mRNA levels of neuropeptides (CGRP, PACAP, VIP) and cytokines (IL-1beta, IL-10) in the trigeminal ganglion, medulla-pons, and hypothalamic area. Of note, the effects of rhBNDF treatment were comparable to those induced by the administration of sumatriptan. rhBDNF administration at both doses significantly reduced serum levels of PACAP, while the higher dose also significantly reduced serum levels of VIP. Conclusions: The findings suggest that intranasal rhBDNF has the potential to be a safe, non-invasive and effective therapeutic approach for the treatment of primary headache, particularly migraine. [ABSTRACT FROM AUTHOR]

Alzheimer's disease (AD), the most common age‐related neurodegenerative condition, is named after Alois Alzheimer and is marked by a progressive deterioration in memory, cognitive function, and behavior. Research has highlighted the importance of nonsteroidal anti‐inflammatory drugs (NSAIDs) in inhibiting the aggregation of amyloid β‐peptide (Aβ), a key feature of AD pathology. Ibuprofen, an NSAID from the propionic acid class, is widely used to manage osteoarthritis and rheumatoid arthritis, exhibiting strong anti‐inflammatory and antipyretic effects. However, the drug's acidic group limits its selectivity for cyclooxygenase (COX) enzymes and contributes to several adverse effects. This study aimed to modify the acidic moiety of ibuprofen into lactone (IBU‐O 1–4) and lactam (IBU‐I 1–3) derivatives to mitigate these side effects. The structural properties of the synthesized imidazolone (IBU‐I 1–3) and oxazolone (IBU‐O 1–4) derivatives were characterized through Q‐TOF LC‐MS, ¹H‐NMR, ¹3C‐NMR, and IR spectroscopy. Molecular docking studies followed by Ellman's method assessed the inhibitory effects of these compounds on acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), while an enzyme immunoassay (EIA) kit was used to evaluate their inhibition of cyclooxygenase‐2 (COX‐2). [ABSTRACT FROM AUTHOR]

The primary issues in treating type 1 diabetes mellitus (T1DM) through the transplantation of healthy islets or islet β-cells are graft rejection and a lack of available donors. Currently, the majority of approaches use cell encapsulation technology and transplant replacement cells that can release insulin to address transplant rejection and donor shortages. However, existing encapsulation materials merely serve as carriers for islet cell growth. A new treatment approach for T1DM could be developed by creating a smart responsive material that encourages the formation of islet cell spheroids to replicate their 3D connections in vivo and controls the release of insulin aggregates. In this study, we used microfluidics to create thermally sensitive porous scaffolds made of poly(N-isopropyl acrylamide)/graphene oxide (PNIPAM/GO). The material was carefully shrunk under near-infrared light, enriched with mouse insulinoma pancreatic β cells (β-TC-6 cells), encapsulated, and cultivated to form 3D cell spheroids. The controlled contraction of the thermally responsive porous scaffold regulated insulin release from the spheroids, demonstrated using the glucose-stimulated insulin release assay (GSIS), enzyme-linked immunosorbent assay (ELISA), and immunofluorescence assay. Eventually, implantation of the spheroids into C57BL/6 N diabetic mice enhanced the therapeutic effect, potentially offering a novel approach to the management of T1DM. [ABSTRACT FROM AUTHOR]

Background: Despite the development of various antiviral drugs, most of them are not effective in the treatment of coronavirus disease 2019 (COVID-19) as a hyperinflammatory disorder. Chemokine (C-C motif) ligand 2 (CCL2) is one of the critical CC chemokines involved in the pathogenesis and severity of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection. This study aimed to investigate the expression of CCL2 and CC chemokine receptor 2 (CCR2) in COVID-19 patients. Methods: Peripheral blood samples were collected from 60 confirmed COVID-19 patients and 60 age-matched healthy subjects. The ages of the subjects were categorized as follows: up to 20 years, 20 to 40 years, 40 to 60 years, and more than 60 years. CCL2 serum levels were measured using the enzyme-linked immunosorbent assay (ELISA). CCR2 gene expression in peripheral blood mononuclear cells (PBMCs) was measured employing real-time polymerase chain reaction (PCR). Results: In all age groups, CCL2 serum levels were significantly elevated in patients compared to healthy controls (P < 0.0001). CCL2 levels were higher in severe patients than in moderate patients. Moreover, CCR2 expression by PBMCs was higher in patients compared to control subjects. However, a significant difference between patients and controls over 60 years of age was identified (P = 0.0353). There was no significant difference in CCR2 expression between moderate and severe COVID-19 patients. Conclusions: Taken together, the findings demonstrate that CCL2 and CCR2 are upregulated in COVID-19 patients at protein and mRNA levels, respectively. Therefore, the CCL2/CCR2 axis may be a potential therapeutic target in order to improve patient outcomes. [ABSTRACT FROM AUTHOR]

Background: To explore the association of long non-coding RNA (lncRNA) SNHG1/ miR-195 axis with type 2 diabetes mellitus (T2DM) and islet function. Methods: The expression of SNHG1 and miR-195 was measured in T2DM patients and in healthy subjects. Correlation between indciators was evaluated using Pearson correlation analysis. INS-1 cells were used to perform the cell function assays. Insulin secretion by INS-1 was detected using ELISA. Cell counting kit-8 (CCK-8) and flow cytometry was used to detect cell proliferation and apoptosis. Luciferase report assay was to used to verify the target of SNHG1. Results: The expression of SNHG1 was increased and miR-195 level was decreased in the serum of T2DM patients. Both SNHG1 and miR-195 could be biomarkers for T2DM diagnosis. The fasting plasma glucose (FPG) and HbA1c were positively related to SNHG1 and negatively related to miR-195. SNHG1 inhibited insulin secretion, and cell proliferation and promoted apoptosis of INS-1 cells via binding to miR-195. Conclusions: Detection of SNHG1 and miR-195 might predict T2DM. SNHG1 could suppress proliferation and insulin secretion, but promote apoptosis of INS-1 cells via sponging miR-195. [ABSTRACT FROM AUTHOR]

Background: We developed the HIV Triplex multiplex bead assay to identify and serotype HIV infection with high sensitivity and specificity; and distinguish recent from long-term HIV-1 infections. It can facilitate accurate incidence estimation, while reducing the number of tests and blood collected, which is highly desirable for use in future studies and surveys. Using previously collected, treatment-naive longitudinal seroconversion HIV-1 positive panels and specimens from individuals infected for >12 months, we determined the assay's mean duration of recent infection (MDRI) and false-recency rate (FRR) respectively, at various mean fluorescent intensity (MFI) cutoffs. Methods: We tested seroconversion specimens (N = 814) from 142 individuals infected with HIV-1 subtypes B, C, or AE, and 1341 cross-sectional specimens from individuals infected >12 months. The MFI cutoffs of 1000 to 2000 were evaluated for recency classification, including an MFI of 1250 corresponding to the limiting antigen avidity enzyme immunoassay (LAg-EIA) cutoff of 1.5 normalized optical density for MDRI and FRR. We used four statistical methods: Methods 1 and 2 used the empirically balanced observation time approach. Method 2 MFI values were raised to power = 1.33, based on a repeated measures model to linearize the relationship between MFI and time points, whereas Method 1 was not linearized. Methods 3 and 4 employed quadratic and linear interpolations for each seroconversion panel. FRR was calculated by dividing the number of specimens misclassified as recent by the total number of specimens tested. Results: MDRI values ranged from 135–146 days at MFI = 1000 to 229–279 days at MFI = 2000 by the 4 methods. FRR varied from 0.15%-1.27% with increasing MFI cutoff. At MFI = 1250, the average MDRI of 4 methods was 169 days and ranged from 159–183 with overlapping 95% CIs and FRR = 0.52%. Conclusion: The HIV Triplex assay demonstrates a longer dynamic range compared to current HIV recency assays with a low FRR for cutoffs examined. With a longer dynamic range and low FRR, the MDRI for recent infection can be extended as appropriate to detect more recent infections, increasing the value of incidence assays benefiting public health surveillance and future surveys. [ABSTRACT FROM AUTHOR]

Effective vaccines against Salmonella Typhi, targeting the Vi capsular polysaccharide, have been developed and are being introduced into routine immunization in endemic countries. Vi conjugated vaccines are also being tested in new multi-component vaccine formulations. Simple, high-throughput and cost-effective assays to quantify Vi-specific IgG in clinical sera are needed. In this study we present the development and qualification of a new anti-Vi ELISA with continuous readout, which expresses results as ELISA Unit/mL (EU/mL). We have qualified the assay in terms of precision, linearity and specificity, demonstrating performance in line with a commercially available anti-Vi ELISA. We have also calibrated the assay against the 16/138 anti-Vi international standard and established conversion factor between EU/mL and international units/mL, to allow comparability of results across studies. In summary, this newassaymet all the suitability criteria and is being used to evaluate anti-Vi responses in clinical studies. [ABSTRACT FROM AUTHOR]