Enzyme-linked Immunosorbent Assay Research Articles

Impact of Malaria and Intestinal Parasitic Infections on Inflammatory Cytokines amongst Armed Forces of the North West and South West, Cameroon

Malaria and intestinal helminthic infections present a significant public health challenge, particularly in tropical regions. The study was designed to determine the impact of Malaria and Intestinal helminthic infections on inflammatory cytokines amongst Armed Forces in the North West and South West Cameroon. This was a cross-sectional study carried out between May 2022 to April 2024. A total of 812 stool and venous blood samples were collected for the identification of malaria and intestinal parasites and for evaluation of inflammatory cytokines. The study used Giemsa staining and Kato-Katz methods to identify malaria and intestinal parasites, while Enzyme-linked immunosorbent assay (ELISA) was used to evaluate inflammatory cytokine concentrations and statistical comparisons were performed using SPSS. Results obtained showed that the prevalence of Malaria, intestinal helminths and co-infections were 13.6%, 22.8% and 3.2% respectively. TNF-α, INF-γ, and IL-1β concentration mean levels were significant in Plasmodium falciparum infected participants (p = 0.000). Trichuris trichiura infected participants showed a positive significant impact on TNF-α (p = 0.000); IL-1β (p = 0.000) and IL-6 (p = 0.035) while Schistosoma mansoni showed a significant impact on TNF-α (p = 0.044) and IL-1β (p = 0.037). Schistosoma intercalatum infected participants showed a significant value of IL-1β (p = 0.004). Also, co-infection with these parasites significantly impacted TNF-α (p = 0.002), INF-γ (p = 0.001), and IL-1β (p =0.000). The study provided evidence that Plasmodium spp., Trichuris trichiura, Schistosoma mansoni, Schistosoma intercalatum and co-infections negatively impacted pro-inflammatory cytokines. This confirms that Malaria and Intestinal helminthic infections engage immunity responses.

Increased expression of proinflammatory cytokines in the cerebrospinal fluid of patients with a history of electrical status epilepticus in sleep.

This study was to investigate the cytokine expression profiles in the cerebrospinal fluid (CSF) of Self-limited epilepsy with centrotemporal spikes (SeLECTS) patients with electrical status epilepticus during sleep (ESES) and compare these to other neurological diseases to identify the cytokines that correlate with ESES. Seven SeLECTS patients with ESES and nine non-ESES controls were included in this study. CSF was centrifuged within one hour and stored at -80°C; subsequently, High mobility group box 1 (HMGB1), Caspase-1, interleukin (IL)-1β, IL-1α, IL-6, IL-8, IL-10 and tumour necrosis factor-α (TNF-α) were measured with enzyme-linked immunosorbent assay. CSF levels of IL-1β, TNF-α, IL-1α, and Caspase-1 in ESES group were significantly elevated compared with those non-ESES controls. Moreover, IL-1β CSF levels were positively correlated with Caspase-1 and TNF-α whereas it moderately correlated with IL-1α. The present study indicates a close relationship between neuroinflammation and ESES pathogenesis in patients with SeLECTS.

Genetic diversity and expanded epidemic area of novel tick-borne pathogen Wetland virus in ticks, wild and domestic animals, and patient in China

Wetland virus (WELV) is a recently identified segmented orthonairovirus associated with human febrile illness in northeastern China. To elucidate its ecological drivers and transmission patterns, we conducted multi-host surveillance in seven eco-climate regions across China through an integrated approach combining molecular and serological profiling. Over 33,000 questing and feeding ticks representing 11 species were systematically screened WELV-RNA alongside 1,493 wild small mammals, while 2,578 domestic animals and 3,921 patients presenting with outdoor field or tick exposure history underwent both WELV specific molecular and serological evaluation. WELV-RNA detection via real-time RT-PCR revealed low but widespread infection, including 0.54% (8/1,493) in wild rodents (Apodemus chevrieri, Rattus norvegicus), 1.12% (29/2,578) in domestic animals, and 0.86% (275/33091) in ticks, including five newly confirmed vector species. Serological analysis via enzyme-linked immunosorbent assay (ELISA) demonstrated 2.44% (63/2578) IgG seroprevalence in livestock. Clinical surveillance identified acute WELV infections in 1.81% (71/3921) of febrile patients, predominantly as monoinfections (77.46%, 55/71). Coinfected patients exhibited elevated disease severity markers, with hospitalization rates doubling compared to single-infection cases (87.50% vs.36.36%). Spatiotemporal analysis showed spring-summer predominance of WELV for both ticks and human patients. Phylogenetic analyses revealed remarkable genetic conservation patterns of WELV strains across diverse hosts and tick vectors; and clustering across geographical regions. These findings establish WELV as an emerging tick-borne virus with trans-regional spread across heterogeneous landscapes in China. The intersecting transmission cycles among wildlife reservoirs, domestic amplifier hosts, and expanding tick vectors underscore the imperative for integrated surveillance strategies to mitigate this growing public health threat.

Assessment of Inflammatory Markers among Hypertensive Women in a Nigerian Population

Objectives Essential hypertension (HTN), a leading cause of preventable death and disability worldwide, remains a major global risk factor for cardiovascular diseases. The study assessed the inflammatory markers of hypertensive women in a Nigerian population. Materials and Methods Two hundred individuals were recruited for this study comprising 80 hypertensive women on treatment, 50 newly diagnosed hypertensive women, and 70 non-hypertensive women (control). Human interferon-gamma (IFN-γ) and transforming growth factor-beta 1 (TGF-β1) as inflammatory markers were analyzed using Enzyme-Linked Immunosorbent Assay. Results The results showed that IFN-γ and TGF-β1 were significantly higher in hypertensive subjects compared with control (P < 0.05). IFN-γ and TGF-β1 were significantly higher (P < 0.05) in hypertensive drug naïve subjects compared with subjects on antihypertensive drugs. Combined use of amlodipine/lisinopril, lisinopril/ metoprolol resulted in significant decrease (P < 0.05) in IFN-γ and TGF-β1 compared with hypertensive patients who used amlodipine and lisinopril alone. There was significant increase (P < 0.05) in IFN-γ and TGF-β1 in age group 51–65 years compared with age group 20–35 years, but there was no significant difference (P > 0.05) in both parameters studied with respect to duration of illness. Conclusion The study concluded that hypertensive subjects undergo inflammation marked by increase in inflammatory markers despite the possible mitigating effect of their antihypertensive medication. Therefore, management of HTN in addition to use of antihypertensive drugs should focus on lifestyle modification, Dietary Approaches to Stop HTN, and increased physical activity.

Exosome-derived circ0009910 promotes pituitary adenoma cell proliferation, invasion, migration, and EMT through the miR-106b-5p/STAT3 axis.

To explore whether exosome-derived circ0009910 can transcellularly regulate the growth of pituitary adenoma cells and to further explore the possible mechanisms of its action. Transmission electron microscopy and nanoparticle size analysis used to observe the morphology and size of the exosomes. Real-time quantitative polymerase chain reaction (qRT‒PCR) used to determine the expression of circ_0009910, miR-106b-5p and STAT3. Western blotting used to assess the expression of exosomal marker proteins, p-STAT3, E-cadherin, N-cadherin and vimentin. Cell Counting Kit-8 (CCK-8) and 5-Ethynyl-2-deoxyuridine(EdU) assays used to determine the proliferative capacity of the cells. Transwell assays were performed to assess the migratory and invasive capacity of the cells. Enzyme-linked immunosorbent assays (ELISAs) used to determine the expression level of GH. A nude mouse xenograft model was established to observe the effects of exosome-derived circ0009910 on transplanted tumors in nude mice. circ0009910 can be transferred to other cells via exosomes and knock down the expression of circ0009910 can inhibit the proliferation, invasion, and migration of pituitary adenoma cells, reduce GH expression, and regulate the expression of epithelial-mesenchymal transition (EMT)-associated proteins; miR-106b-5p is a molecular sponge of circ0009910 and can partially reverse its procarcinogenic effect of circ0009910 in pituitary adenoma, and STAT3 is a target gene of miR-106b-5p. In addition, circ0009910 knockdown inhibited tumor growth in vivo. Exosome-derived circ0009910 promotes pituitary adenoma progression and regulates EMT through the miR-106b-5p/STAT3 axis.

Positive Impact of Holistic Nursing on Cognitive Impairment and Psychiatric Symptoms in Patients With Alzheimer's Disease.

Alzheimer's Disease (AD) affects millions of elderly individuals worldwide and has been clinically recognized as one of the most significant disorders compromising quality of life in late-stage human development. The objective of this study is to systematically evaluate the influence of holistic nursing (HN) on patients with AD, thereby providing evidence-based references for clinical practice. A total of 105 patients with AD hospitalized in our hospital between January 2023 and January 2024 were enrolled for prospective analysis. Among them, 58 received conventional care (control group), and 47 received HN (observation group). Before and following the nursing interventions, both groups underwent assessment using the Mini-Mental State Examination (MMSE), Neuropsychiatric Inventory (NPI), Alzheimer's Disease Assessment Scale-Cognitive Subscale (ADAS-cog), Montreal Cognitive Assessment (MoCA), and Self-rating Anxiety/Depression Scales (SAS/SDS). In addition, neurotransmitter levels and neuroinflammatory markers were measured using enzyme-linked immunosorbent assay and fully automated chemiluminescent immunoassay. Treatment compliance, incidence of adverse events, and family satisfaction were also recorded and compared between the two groups. After nursing interventions, the observation group demonstrated significantly higher MMSE and MoCA scores compared to the control group. Conversely, NPI, ADAS-cog, SAS, and SDS scores were notably lower in the observation group (p < 0.05). Furthermore, neurotransmitter levels were significantly elevated in the observation group, while the concentrations of central nervous system-specific protein β (S100β) and interleukin-1β (IL-1β) were significantly reduced (p < 0.05). Although the incidence of adverse events did not differ significantly between the two groups (p > 0.05), the observation group exhibited higher treatment compliance and greater family satisfaction (p < 0.05). HN effectively improves cognitive function and alleviates psychiatric symptoms in AD patients, supporting its recommendation for clinical application. No. NCT06868004.

Compatibility between Proteins and Polysaccharide Excipients in Oral Delivery Tablets.

The development of oral protein biopharmaceuticals has gradually become a hot area of research. Currently, most of the added excipients are oral excipients used in traditional small molecule drugs, such as starch and lactose. However, the interaction between proteins and oral excipients has not received much attention. For example, the interaction may have an effect on protein stability, efficacy, etc. Herein, we observed a strong interaction between protein and sodium carboxymethyl starch (CMS), a common disintegrator in oral formulations of small molecule drugs. CMS can cause the complete disappearance of a free soluble protein in trastuzumab (TRA) tablets after reconstituted, as detected by size-exclusion high-performance liquid chromatography. While the other two polysaccharides with similar structures, hydroxyethyl starch and methylcellulose, exhibited no effect on the soluble TRA concentration. CMS produces an electrostatic interaction with TRA in a certain pH range, which affects the soluble protein monomer concentration and solution turbidity, increases the hydrophobicity of the TRA-CMS complex, and reduces the thermal stability of TRA, without changing its biological activity detected by an enzyme-linked immunosorbent assay. Thus, the CMS and TRA electrostatic binding is reversible, and CMS has a potential application in sustained release in oral monoclonal antibody (mAb) and protein delivery tablets. Mixed solutions of infliximab and etanercept (a fusion protein) with CMS at different pH were also studied by SE-HPLC. The results showed that the concentration of the free soluble protein detected by SE-HPLC was significantly and universally reduced when pH was below the isoelectric point, where the positively charged protein forms a strong electrostatic interaction with the negatively charged CMS. The structural characteristics of the protein (such as surface charge, surface hydrophobicity, etc.) should be fully considered in selecting the appropriate excipients for protein biopharmaceutical tablets.

Performance of a whole blood immunoassay for tenofovir detection and correlation with self-reported pre-exposure prophylaxis use in HIV-negative men who have sex with men interested in blood donation.

In 2023, the United States Food and Drug Administration revised its blood donor eligibility policy for men who have sex with men (MSM) from a 3-month deferral to individual assessment. Human Immunodeficiency Virus (HIV) pre-exposure prophylaxis (PrEP) use remains a reason for deferral, and nondisclosure is a concern. In a cross-sectional study of sexually active MSM from 8 U.S. cities who were interested in future blood donation, we assessed the performance of an enzyme-linked immunosorbent assay for detecting tenofovir (TFV) in whole blood (WB) and plasma and the correlation with self-reported PrEP use. Of 1548 individuals, 48% reported oral PrEP use. The WB assay identified 95% of PrEP users, while the plasma assay detected 88%. The WB assay performed well up to 14 days after the last reported dose. Receiver operating characteristics curve analysis showed an area under the curve of 0.96 (95% confidence interval [CI]: 0.95-0.97) using WB and 0.88 (95% CI: 0.86-0.90) using plasma. Specificity was 80% for WB and 66% for plasma. Detection rates for TFV disoproxil fumarate/emtricitabine (FTC) formulations were 99% in WB and 98% in plasma, compared to 93% and 86% for the TFV alafenamide/FTC formulation. High concordance between self-reported oral PrEP use and TFV detection was observed among PrEP users, suggesting the potential utility of WB as a biomatrix for TFV detection to support screening strategies. Given the expanded eligibility for MSM, who may be PrEP users, to donate blood, further examination of undisclosed PrEP use is important.

Recombinase Polymerase Amplification (RPA)-ELISA as an Isothermal Molecular POCT Method for Bacterial Respiratory Infection Diagnosis

Background: Acute Respiratory Infections (ARIs) are a leading cause of childhood mortality worldwide, especially in African and Southeast Asian countries. Point of Care Test (POCT) techniques provide faster diagnoses compared to conventional or real-time PCR methods. Recombinase Polymerase Amplification (RPA) offers rapid on-site detection of these infections. Coupling RPA with Enzyme-Linked Immunosorbent Assay (ELISA) (RPA-ELISA) creates a cost-effective alternative, ideal for clinical applications. This study evaluates RPA-ELISA as a rapid diagnostic tool for bacterial respiratory infections. Methods: From 11 August 2022 to 9 February 2023, respiratory samples were collected and processed using culture methods, biochemical tests, real-time PCR, and RPA assays. The RPA reactions were conducted at 39°C for 30 min, and ELISA was used for detection. Statistical analyses focused on sensitivity, specificity, Positive Predictive Values (PPV), and Negative Predictive Values (NPV). Results: Forty-two respiratory samples, were collected in this period of which 10 samples showed no growth, and 32 tested positive. Among these positive samples, 15 isolates (35.7%) were identified as Klebsiella pneumoniae (K. pneumoniae), 14 isolates (33.3%) as Streptococcus pneumoniae (S. pneumoniae), and 3 isolates (7.1%) as Moraxella catarrhalis (M. catarrhalis). RPA-ELISA demonstrated 100% sensitivity for all pathogens, comparable to or better than RT-PCR, but had slightly lower specificity and PPV. RT-PCR achieved 100% specificity and PPV for all pathogens, indicating higher accuracy; yet, RPA-ELISA's sensitivity points to its effectiveness as a rapid screening tool. Conclusion: RPA-ELISA is significantly faster than real-time PCR and culture methods. Its ease of use makes it suitable for on-site diagnoses in resource-limited environments. Limitations include a small sample size for certain bacteria and the necessity for further validation in varied clinical contexts.

Investigating the effects of 40 Hz sound stimulation on Alzheimer's disease pathways: Modulation of amyloid-β42 secretion, tau phosphorylation, phagocytosis, and autophagy.

BackgroundAlzheimer's disease (AD) is the main cause of dementia in an aging society. Previous studies have demonstrated that non-invasive light flicker and sound with gamma frequency oscillations can modulate AD-related pathology in AD mice, potentially improving patient outcomes. However, the molecular mechanism by which sound with gamma frequency oscillations inhibits the expression of amyloid-β1-42 (Aβ42) and the phosphorylation of tau, and modulating cell autophagy in nerve cells are still unclear.ObjectiveThis study aimed to explore the molecular effects of 40 Hz sound stimulation on AD-related pathways in a cellular model.MethodsWe designed a 40 Hz stimulating sound (H+ multi-frequency audio) for this study, and cells were exposed to H+ multi-frequency audio. The concentration of Aβ42 was quantified by enzyme-linked immunosorbent assay. Protein levels were examined by western blotting. Phagocytosis was examined by confocal microscopy and phagocytic analysis.ResultsFirst, we found that exposure to the 40 Hz stimulating sound inhibited the secretion of Aβ42 by activating the AβPP/ADAM10 pathway and suppressing the AβPP/BACE1 pathway. Second, 40 Hz stimulating sound inhibited tau phosphorylation at Thr181 through the inactivation of the Akt/mTOR pathway. Third, 40 Hz stimulating sound enhanced the phagocytosis and autophagy of Aβ42 through the AMPK/ULK/LC3B pathway in cells.ConclusionsOur study showed that 40 Hz stimulating sound is involved in the inhibition of Aβ42 secretion, p-Tau protein expression, and the promotion of phagocytosis and Aβ42 autophagy in cells. We suggest that 40 Hz stimulating sound could be a potential intervention to attenuate AD progression in the future.

Assessment of Serum Cytokine Levels in Keratoconus Patients

Background: While keratoconus (KC) has traditionally been classified as a non-inflammatory corneal disorder, emerging evidence suggests that inflammatory processes may be involved in its pathogenesis. This study investigated whether systemic inflammation contributes to KC development by measuring serum levels of key pro-inflammatory cytokines in KC patients compared to healthy controls. Methods: This cross-sectional comparative study included 60 participants aged 18–30 and was divided into three equal groups: healthy controls, progressive KC, and stable KC. The levels of interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and interleukin-1 beta (IL-1β) in the serum were measured through an enzyme-linked immunosorbent assay. KC severity was classified based on mean keratometry values. Results: There were no significant differences detected in serum IL-6, TNF-α, and IL-1β levels across groups (p &gt; 0.05). Moreover, no significant correlations were found between systemic cytokine levels and KC severity categories (p &gt; 0.05). Conclusions: Systemic levels of IL-1β, IL-6, and TNF-α do not significantly differ between KC patients and controls nor do they correlate with disease severity, reinforcing the hypothesis that KC involves primarily local inflammatory processes.

Traditional Chinese medicine Youguiyin decoction ameliorate glucocorticoid-induced osteonecrosis in rat by modulating ROS/PHD2/HIF-1α oxidative stress signaling pathway in bone marrow mesenchymal stem cells

BackgroundThe incidence of osteonecrosis is increasing annually due to the widespread use of glucocorticoids. Recent evidence suggests a significant association between glucocorticoid-induced osteonecrosis and oxidative stress. Youguiyin (YGY) decoction, a classic formula of traditional Chinese medicine, has been widely used for the prevention of glucocorticoid-induced osteonecrosis. However, its underlying pharmacological mechanisms are still not fully understood.MethodsUPLC-Q-TOF–MS and network pharmacology were used to elucidate the material basis of YGY decoction and its mechanism for the treatment of glucocorticoid-induced osteonecrosis. The anti-oxidative stress and bone-enhancing effects in vivo were detected by hematoxylin–eosin (HE) staining, serum metabolomics, enzyme-linked immunosorbent assay (ELISA), immunohistochemistry (IHC), and Western Blot (WB). Rat bone marrow mesenchymal stem cells (BMSCs) were induced with dexamethasone (DXMS) for 24 h, followed by YGY medicated serum for 24 h. Significantly up- and down-regulated genes were detected by RNA sequencing. Oxidative stress levels were detected by ROS fluorescence. Alizarin red S staining was used to detect osteogenic effects. WB and ELISA were used to detect the expression of proteins related to the ROS/PHD2/HIF-1a pathway.ResultsThe application of YGY decoction significantly promoted bone repair and antagonized excess reactive oxygen species (ROS) generation in glucocorticoid-associated osteonecrosis of the femoral head (GA-ONFH) rats. In addition, YGY medicated serum antagonized DXMS-induced ROS production and promoted osteogenic differentiation in BMSCs. We also found that YGY medicated serum attenuated excess ROS generation while PHD2 expression was significantly increased, HIF-1α expression was significantly decreased and RUNX2 expression was significantly increased.ConclusionThese results provide compelling in vivo and in vitro evidence that YGY decoction may play a role in promoting glucocorticoid-induced osteonecrosis bone repair by targeting the mediation of the ROS/PHD2/HIF-1α oxidative stress signaling pathway, thus providing a new theoretical basis for the clinical application of YGY decoction to glucocorticoid-induced osteonecrosis.Graphical

Investigating the Therapeutic Potential of Crude Leech Saliva Based on Its Anticancer, Antioxidant, and Anti-Inflammatory Effects

Leech therapy is a biotherapeutic approach that has been traditionally used for centuries and is currently being re-evaluated in modern medicine. The efficacy of this treatment is attributed to various bioactive compounds found in leech saliva, which exhibit anticoagulant, anti-inflammatory, antioxidant, and anticancer properties. It has been demonstrated that leech saliva possesses the potential to modulate inflammatory processes and apoptotic mechanisms. In this study, the therapeutic potential of the saliva of Hirudo verbana was evaluated, and its biological and pharmacological effects were comprehensively investigated. The anticancer effects, antioxidant capacity, and anti-inflammatory activity of the crude leech saliva were assessed using human umbilical vein endothelial cells and epithelial ovarian cancer cells. The chemical composition of the saliva was analyzed using gas chromatography–mass spectrometry, while the protein content was determined by the Bradford assay. Antioxidant activity was measured using the 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay, inflammatory effects were evaluated by Enzyme-Linked ImmunoSorbent Assay, and cell viability was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The findings revealed that crude leech saliva had a minimal effect on healthy cells but showed a selective effect on the viability of ovarian cancer cells. At low concentrations (3.13%), 99.16% of healthy cells remained viable, whereas this rate decreased to 89.25% in cancer cells; at high concentrations (50%), cell viability in cancer cells declined to 63.02%. Gas chromatography–mass spectrometry analysis identified compounds such as gibberellic acid and 6-[(4-methoxyphenyl)methoxy]-4,4,5,7,8-pentamethyl-3H-chromen-2-one, which demonstrated high affinity for the antiapoptotic proteins Bcl-2 and Survivin in molecular docking analyses. In conclusion, the crude leech saliva was confirmed to possess anti-inflammatory, antioxidant, and anticancer properties. However, further biochemical and clinical research is needed to elucidate the underlying mechanisms of these biological effects in greater detail.

Obacunone acts as a histone deacetylase 1 inhibitor to limit p38MAPK signaling and alleviate osteoarthritis progression

BackgroundOsteoarthritis (OA) is an age-related progressive degenerative disorder characterized by cartilage extracellular matrix degradation and inflammation. In this study, we explored the function and mechanism of action of obacunone (OB) in inhibiting OA progression.MethodsThe degradation of articular cartilage and its severity were examined using Safranin O-fast green and hematoxylin and eosin (HE) staining. Chondrocyte survival was evaluated using a cell counting kit-8 assay. In addition, qRT-PCR, western blot analysis, immunohistochemical staining, and enzyme-linked immunosorbent assay were performed to evaluate the effects of OB on cartilage injury.ResultsOB mitigated cartilage lesions in rats with anterior cruciate ligament transaction-induced OA. The protein expression of collagen II was increased and the protein expression of ADAM metallopeptidase with thrombospondin type 1 motif 5 (ADAMTS-5), matrix metalloproteinase (MMP)-13, and RUNX family transcription factor 2 (RUNX2) was reduced in the articular cartilage of OB-treated rats. Moreover, OB exhibited anti-inflammatory activities by reducing the serum levels of interleukin (IL)-6, tumor necrosis factor (TNF)-α, IL-1β, and IL-18. In IL-1β-stimulated primary chondrocytes, OB dose-dependently elevated the expression of collagen II, and decreased the expression of ADAMTS-5, MMP-13, RUNX2 and inflammatory cytokines. Histone deacetylase 1 (HDAC1) was identified as a predicted OB target. OB inhibited HDAC1 expression to limit the activation of p38MAPK signaling. The transfection of chondrocytes with HDAC1 or p38MAPK overexpression plasmids reversed the chondroprotective effects of OB.ConclusionOB mitigated OA progression by binding to HDAC1 and inhibiting p38MAPK signaling, indicating that OB may be a promising drug for the treatment of OA.

Aerobic Exercise and Celastrol: A Promising Combination for Achilles Tendon Regeneration

Background: Tendons are subjected to mechanical stress and strain during daily activities and physical exercise, which makes them susceptible to injury. Objectives: This randomized trial evaluated the effects of a 4-week combination of training and celastrol on the healing of Achilles tendon ruptures in rats. It assessed cytokine levels, gene expression, and histological changes. Methods: Rats with Achilles tendon ruptures were randomly assigned to several treatment groups: Sham, negative control, training, celastrol, and a combination of training and celastrol. Eligibility criteria included age, weight, and the absence of pre-existing tendon injuries. The assessments included enzyme-linked immunosorbent assay (ELISA) to measure cytokine levels [bFGF, IGF1, cyclooxygenase 2 (COX2)], real-time polymerase chain reaction (RT-PCR) to analyze gene expression (scleraxis, periostin, MMP9, collagen I), and histological analysis using Masson's trichrome staining to evaluate collagen organization. Results: The training-celastrol group exhibited the highest levels of bFGF, IGF1, and COX2, indicating a synergistic effect on tendon healing compared to the sham and negative control groups, with effect sizes of 0.85 (95% CI: 0.70 - 1.00). Additionally, RT-PCR results showed that the training-celastrol group had the highest expression of scleraxis, periostin, MMP9, and collagen I, suggesting enhanced tendon repair and remodeling. Histological analysis revealed that the training-celastrol group displayed the most organized collagen fibers, whereas the sham group showed regular organization, and the negative control group exhibited disorganized fibers. Conclusions: The study concluded that combining aerobic training and celastrol significantly enhances tendon healing by improving cytokine levels, gene expression, and collagen organization. Outcome measures were assessed at baseline, mid-treatment (week 2), and post-treatment (week 4) to evaluate temporal changes. These findings highlight the effectiveness of these therapeutic strategies in promoting tendon repair and maintaining structural integrity.

The role of complement C1q in atrial fibrillation: a marker for disease progression and surgical outcomes

Background and objectiveThe complement system plays a crucial role in the pathogenesis and progression of cardiovascular diseases. C1q, a key initiator of the classical pathway, is closely associated with various chronic inflammatory conditions. This observational study aims to elucidate the potential risk relationship between serum complement C1q levels and atrial fibrillation (AF).Materials and methodsThis retrospective cohort study included 812 AF patients treated at the Affiliated Hospital of Qingdao University from January 2020 to October 2022, comprising 694 patients in the paroxysmal AF group and 118 in the persistent AF group. Serum complement C1q levels were measured using an enzyme-linked immunosorbent assay (ELISA).ResultsSerum C1q levels in the AF group were significantly lower than those in the control group (P &amp;lt; 0.001). Logistic regression analysis indicated that reduced plasma C1q levels were independently associated with the incidence of AF (95% CI = 0.974–0.981, P = 0.001). Additionally, ROC curve analysis confirmed the close association between plasma C1q levels and AF, highlighting the predictive value of C1q for AF. Further investigation revealed that C1q serves as an independent risk factor for complex fractionated atrial electrograms (CFAE) in the superior left atrium of paroxysmal AF patients (95% CI = 0.984–0.998, P = 0.031), suggesting its potential as a clinical indicator for guiding AF surgical interventions.ConclusionSerum C1q levels are significantly reduced in patients with AF. The presence of CFAE in the superior left atrium of paroxysmal AF patients may be potentially associated with C1q levels. Low complement levels are associated with atrial fibrillation compared to individuals without AF and may represent a potential underlying cause of impaired sinus rhythm maintenance following pulmonary vein isolation. Complement C1q may play a critical role in the pathogenesis of AF.

Higher Hepatitis E Seroprevalence in Workers in the Pig Sector in Flanders, Belgium: Results From a Seroprevalence Case-Control Study.

Autochthonous hepatitis E virus (HEV) infection in industrialised countries is a zoonosis. Pigs are recognised as the principal reservoir. Consequently, workers in the pig sector may be a high-risk occupational group for HEV infection. The current study aims to investigate whether workers exposed to pigs or pork have a higher seroprevalence of HEV IgG in comparison to a control group. A cross-sectional study was conducted involving 92 individuals occupationally exposed to pigs or pork and 217 control subjects outside the pig sector. Socio-demographic variables, information about occupation, and a number of relevant confounding variables (such as eating habits, and history of blood transfusions) were gathered by a questionnaire. Serum samples were tested for anti-HEV IgG antibodies using an enzyme-linked immunosorbent assay (ELISA). Our study revealed a significantly higher seroprevalence of HEV antibodies in pig workers (32.6%) compared to the control group (9.2%) (p < 0.001). The relation between occupational exposure and HEV seropositivity remained significant in the multiple logistic regression analysis, in which the odds were adjusted for age, gender, and eating habits. The results from this first HEV IgG serology study in the pig sector in Belgium demonstrate that workers in this sector have a higher risk for HEV infection compared to a population not occupationally exposed to pigs and pork. These findings underscore the need for targeted occupational health interventions, including improved hygiene practices and informing employees with risk profiles, to mitigate the risk of HEV infection among pig workers.

Extracellular vesicles therapy alleviates cisplatin-ınduced testicular tissue toxicity in a rat model.

Cisplatin is a commonly used chemotherapy agent effective against various cancers, however it induces significant gonadotoxicity and infertility due to its adverse effects on testicular function. The underlying mechanisms of cisplatin-induced testicular damage include oxidative stress and dysregulated autophagy. This study investigates the potential of extracellular vesicles (EVs) to mitigate cisplatin-induced testicular damage through their regenerative, antioxidant, and autophagy-modulating properties. In the testicular toxicity model, thirty-two male rats were randomly divided into four groups (n = 8): control, EVs-only, Cis-only, and Cis + EVs. A single intraperitoneal dose of 7.5mg/kg cisplatin was administered on the first day. On the six day, the EVs treatment group received a single dose of EVs (8x107/100μl) intravenously. Animals were sacrificed on day eight. Testicular histoarchitecture was assessed via hematoxylin and eosin staining. Sperm parameters, including motility and count, were measured using light microscopy. Hormone levels (testosterone and inhibin) were determined via enzyme-linked immunosorbent assay (ELISA). Oxidative stress markers, such as glutathione peroxidase (GSH-PX), superoxide dismutase (SOD), catalase (CAT), and is a metabolite malondialdehyde (MDA), were quantified using colorimetric assays. Autophagy and steroidogenesis were evaluated through immunohistochemical analysis of Beclin-1, p62, LC3-2, SF-1, and StAR. Cisplatin exposure caused significant testicular damage, characterized by reduced germinal epithelium and degeneration of seminiferous tubules (p < 0.001). These structural changes led to hormonal imbalances, as evidenced by declines in testosterone (p < 0.005) and inhibin (p < 0.001). Additionally, sperm motility (p < 0.05) and count (p < 0.001) were adversely affected. Immunohistochemical analysis revealed upregulation of autophagy markers (p < 0.001), indicating heightened autophagic activity, alongside downregulation of steroidogenic factors (p < 0.001), which contributed to impaired steroidogenesis. Elevated levels of malondialdehyde (MDA) (p < 0.01) and decreased activities of antioxidant enzymes-GSH-PX, SOD, and CAT (p < 0.001) pointed to increased oxidative stress as a contributing mechanism. In contrast, treatment with extracellular vesicles (EVs) significantly improved testicular histoarchitecture (p < 0.001) and restored hormonal levels toward normal (testosterone p < 0.005, inhibin p < 0.001). Furthermore, EVs reduced the expression of autophagy markers (p < 0.001) and enhanced the levels of steroidogenic factors (p < 0.05). Notably, MDA levels decreased (p < 0.001), while antioxidant activities increased (p < 0.001), suggesting a protective effect of EVs against oxidative stress. EVs protect against cisplatin-induced reproductive toxicity by modulating oxidative stress and autophagy pathways, preserving testicular function and fertility. These findings suggest that EVs may be a promising therapeutic strategy for mitigating cisplatin's negative effects on reproductive health. Further exploration of dosing regimens and localized applications is recommended for improved efficacy.

Inverted day-night feeding during pregnancy affects the brain health of both maternal and fetal brains through increasing inflammation levels associated with dysbiosis of the gut microbiome in rats

BackgroundIn both humans and rodents, inappropriate feeding times during pregnancy can cause maternal metabolic abnormalities, increasing the risk of neurodevelopmental disorders in both the mother and offspring. Using a rat model, this study investigates whether feeding only during the inactive phase in rats leads to anxiety-like behaviors and abnormal brain development in fetuses through gut microbiota imbalance.Methods10-week-old female rats in the inactive-phase feeding group (IF group) were first trained for daytime feeding, ensuring that energy intake was statistically insignificant and different from that of the normal diet feeding group (ND group). They were then paired with male rats, and the previous feeding regimen was continued after pregnancy. Anxiety-like behavior was evaluated using the open-field test. Maternal caecal microbiota was analyzed using 16S rRNA sequencing. Enzyme-linked immunosorbent assay (ELISA) measured serum inflammation factors. RT-PCR was employed to assess mRNA levels of integrity genes and inflammatory cytokines in the maternal hippocampi, intestines, fetal brains, and placentae.ResultsThere were no statistically significant differences in energy intake or body weight gain between the IF and ND groups. In the open field test, dams in the IF group exhibited anxiety-like behavior, as indicated by fewer entries into and shorter duration in the central zone. Active-phase fasting elevated maternal serum inflammatory cytokine levels and impaired antioxidant capacity. It also increased intestinal permeability and induced gut microbiota dysbiosis, characterized by a decrease in Akkermansia and an increase in Dubosiella. Changes in the expression of intestinal circadian genes and elevated intestinal inflammatory cytokines were observed. Lipopolysaccharide (LPS) translocated into the maternal circulation, activated Toll-like receptor 4 (TLR 4), and passed through the compromised placental barrier into the fetal brain, leading to increased expression of inflammatory cytokines in the fetal brain.ConclusionsThe misalignment between maternal feeding time and the biological clock during pregnancy disrupts the balance of the gut microbiota and peripheral rhythms. The impaired intestinal and placental barriers allow LPS from the gut to infiltrate the maternal hippocampus and fetal brain, increasing inflammation and impacting both maternal and fetal brain health.

Surrogate Markers of Intestinal Permeability, Bacterial Translocation and Gut-Vascular Barrier Damage Across Stages of Cirrhosis.

Portal hypertension, gut barrier dysfunction, and pathological bacterial translocation are hallmarks of cirrhosis driving complications. As measuring gut barrier function is demanding, surrogate markers have been proposed, but their intercorrelation and applicability across different stages of advanced liver disease, particularly in acute-on-chronic liver failure (ACLF), are largely unknown. Proposed markers of gut barrier dysfunction and bacterial translocation were quantified in sera from 160 patients with cirrhosis across different disease stages of compensated and decompensated cirrhosis as well as from 20 patients in hepatic and portal vein serum before and after the insertion of transjugular intrahepatic portosystemic stent (TIPS) using enzyme-linked immunosorbent assay (ELISA). Across all stages of liver disease, the gut-vascular barrier (GVB) marker plasmalemma vesicle protein-1 (PV-1) correlated with bacterial translocation markers endogenous endotoxin-core IgA antibodies (EndoCAb) and LPS-binding protein (LBP) but not with intestinal damage markers intestinal fatty acid binding protein (I-FABP) and zonulin-family peptides (ZFP). PV-1 and EndoCAb were higher in decompensated cirrhosis without further increase in ACLF. Among investigated markers, only I-FABP correlated with the portosystemic pressure gradient, and TIPS insertion significantly reduced portal concentrations within 24 h. Higher PV-1 levels indicated poor transplant-free survival in univariate and multivariable analysis. Surrogate markers of bacterial gut barrier dysfunction and bacterial translocation like ZFP, LBP and EndoCAb appear of limited use in advanced stages of cirrhosis and are confounded by hepatic synthesis capacity, portal congestion and acute phase responses. The prognostic implications of circulating PV-1 in decompensated cirrhosis levels demand further investigation.