Enzyme-linked Immunosorbent Assay Research Articles

Intervention effects of Naoxintong capsules on psychological and cardiac status in depressed rats after heart failure

Abstract Background Depression is a common clinical phenomenon in the patients with heart failure (HF). In traditional Chinese medicine (TCM), diseases in the brain and heart are thought to be correlated and interact. Naoxintong capsules (NXT) has been used for treating cardio-cerebrovascular diseases, while its therapeutic effect on depression after HF remains unclear. Objective The aim of the study is to evaluate the intervention effect of NXT on depression after HF. Methods Sprague-Dawley rats were assigned into the following 5 groups: sham, model, NXT (250, 1000 mg/kg), and valsartan (8 mg/kg). Coronary artery occlusion was performed to induce HF and subsequent depression in rats. The cardiac function was evaluated by echocardiography, hematoxylin-eosin staining, and Masson trichrome staining. The sucrose preference test and Morris water maze test were carried out to assess the depressive behaviors in rats. The ultrastructure of hippocampal CA1 neurons was observed and the levels of corticotropin-releasing hormone in the hypothalamus, brain-derived neurotrophic factor in the cortex, and adrenocorticotropic hormone (ACTH) in the plasma were determined by enzyme-linked immunosorbent assay. The levels of dopamine, 5-hydroxytryptamine, norepinephrine, and γ-aminobutyric acid in the hippocampus were measured by UPLC-QQQ-MS. Results NXT reduced myocardial injury and pathological changes in the cardiac tissue and increased the left ventricular ejection fraction, left ventricular fractional shortening, and cardiac output. NXT increased the sugar preference rate and number of crossings and shortened the escape latency. Furthermore, the NXT treatment restored the levels of corticotropin-releasing hormone, adrenocorticotropic hormone, brain-derived neurotrophic factor, dopamine, and γ-aminobutyric acid to the baseline values. Conclusions NXT not only demonstrates cardioprotective effect but also attenuates depression in the rats after HF. It may exert the antidepressant effect by inhibiting the hyperactivity of the hypothalamic-pituitary-adrenal axis and recovering the levels of neurotrophic factors and neurotransmitters.

Biochemical markers and carotid intima-media thickness in relation to cardiovascular risk in young women.

Cardiovascular disease (CVD) is a major cause of death in the female population. The current study aimed to examine the relationship between CVD risk and novel endothelial dysfunction biomarkers [i.e., endocan, adiponectin and intercellular adhesion molecule (ICAM)-1] and carotid intima-media thickness (cIMT), respectively in a cohort of disease-free women of reproductive age. A total of 129 women were selected. Serum endocan, adiponectin and ICAM-1 were measured by a commercial enzyme-linked immunosorbent assay and cITM was determined by ultrasound. Cardiovascular risk score (CVRS) was calculated. The lowest endocan (p for trend = 0.051) and adiponectin (p for trend = 0.040) levels were found in a group of subjects with the highest CVRS. The cIMT values were the highest in second tertile subgroups, with the highest 75th percentile in a third tertile CVRS group, while the lowest cIMT values were detected in the lowest CVRS tertile group (p for trend = 0.001). A significant positive correlation between cIMT and CVRS (ρ = 0.307, p < 0.001), and a negative correlation between adiponectin and endocan with CVRS, respectively (ρ = - 0.252, p = 0.004; ρ = - 0.179, p = 0.043) were observed, but only endocan retained the independent association with CVRS (p = 0.030) in the multiple linear regression analysis. Endocan could be useful diagnostic tool in the estimation of cardiovascular risk in young women.

Influence of Lactation Stage on Content of Neurotrophic Factors, Leptin, and Insulin in Human Milk.

Human milk comprehensively meets the nutritional needs of a child, providing not only structural and energy components but also various bioactive factors. Among these, neurotrophic factors and hormones involved in metabolic processes deserve special attention. Studies using enzyme-linked immunosorbent assays compared the content of neurotrophic factors-CNTF, NT-3, and NGF-and hormones, leptin and insulin, in two groups of breast milk samples: early lactation (1-3 months) and extended lactation (>6 months, up to 12 months). The results indicated changes in leptin and insulin levels as the lactation period extended. NGF, leptin, and insulin were present in milk samples from both study groups, with leptin and insulin levels being higher in the early lactation group. CNTF and NT-3 were not detected in any of the samples from either study group. The analyses confirmed that human milk from women who breastfeed for extended periods remains a source of biologically active components and macronutrients that support a child's development and health.

Enhancing Th17 cells drainage through meningeal lymphatic vessels alleviate neuroinflammation after subarachnoid hemorrhage

BackgroundSubarachnoid hemorrhage (SAH) is a severe cerebrovascular disorder primarily caused by the rupture of aneurysm, which results in a high mortality rate and consequently imposes a significant burden on society. The occurrence of SAH initiates an immune response that further exacerbates brain damage. The acute inflammatory reaction subsequent to SAH plays a crucial role in determining the prognosis. Th17 cells, a subset of T cells, are related to the brain injury following SAH, and it is unclear how Th17 cells are cleared in the brain. Meningeal lymphatic vessels are a newly discovered intracranial fluid transport system that has been shown to drain large molecules and immune cells to deep cervical lymph nodes. There is limited understanding of the role of the meningeal lymphatic system in SAH. The objective of this research is to explore the impact and underlying mechanism of drainage Th17 cells by meningeal lymphatics on SAH.MethodsTreatments to manipulate meningeal lymphatic function and the CCR7-CCL21 pathway were administered, including laser ablation, injection of VEGF-C geneknockout, and protein injection. Mouse behavior was assessed using the balance beam experiment and the modified Garcia scoring system. Flow cytometry, enzyme-linked immunosorbent assays (ELISA), and immunofluorescence staining were used to study the impact of meningeal lymphatic on SAH drainage. Select patients with unruptured and ruptured aneurysms in our hospital as the control group and the SAH group, with 7 cases in each group. Peripheral blood and cerebrospinal fluid (CSF) samples were assessed by ELISA and flow cytometry.ResultsMice with SAH showed substantial behavioral abnormalities and brain damage in which immune cells accumulated in the brain. Laser ablation of the meningeal lymphatic system or knockout of the CCR7 gene leads to Th17 cell aggregation in the meninges, resulting in a decreased neurological function score and increased levels of inflammatory factors. Injection of VEGF-C or CCL21 protein promotes Th17 cell drainage to lymph nodes, an increased neurological function score, and decreased levels of inflammatory factors. Clinical blood and CSF results showed that inflammatory factors in SAH group were significantly increased. The number of Th17 cells in the SAH group was significantly higher than the control group. Clinical results confirmed Th17 cells aggravated the level of neuroinflammation after SAH.ConclusionThis study shows that improving the drainage of Th17 cells by meningeal lymphatics via the CCR7-CCL21 pathway can reduce brain damage and improve behavior in the SAH mouse model. This could lead to new treatment options for SAH.

Association between vitamin D levels with IL-6 and IL-10 in umbilical cord blood of infants

A worldwide issue, vitamin D deficiency affects pregnant mothers and babies everywhere, including Indonesia. It involves the adaptive immune system by controlling the production of pro- and anti-inflammatory cytokines and the balance between humoral (Th2) and cell-mediated (Th1) immunity. The aim of this study was to investigate the relationship between vitamin D and the cytokines IL-6 and IL-10 in infants. It also examined the relationship between ferritin and IL-6/IL-10 in newborns. The study collected 114 umbilical cord blood samples from term-born mothers without clinical symptoms. IL-6 and IL-10 were among the cytokine profiles measured by the enzyme-linked immunosorbent assay (ELISA). SPSS was used for statistical analysis, and an in-silico investigation was carried out to examine the molecular relationships between vitamin D and IL-6/IL-10. Using the 20 ng/mL as the cut-off for vitamin D insufficiency suggested the insignificant association of vitamin D with IL-6 (p=0.42), IL-10 (p=0.76), and ferritin (p=0.47). When the umbilical cord vitamin D level was categorized into four quartiles, the association with the highest statistical significance (quartile 4 versus quartile 2) was observed for IL-6 (p&lt;0.001), IL-10 (p&lt;0.001), and ferritin (p&lt;0.001). However, the linear regression did not suggest the significant correlations of vitamin D with IL-6 (p=0.40) and IL-10 (p=0.45). A significant correlation based on the linear regression was found between ferritin and IL-10 (p=0.03). Molecular docking studies demonstrated binding affinities of -8.04 kcal/mol for IL-6-vitamin D and -8.53 kcal/mol for IL-10-vitamin D complexes, with stable root mean square deviation throughout the simulations. This study contributes valuable insights into the clinical and computational analysis of the relationship of vitamin D with IL-6 or IL-10.

RGS10 Deficiency Alleviated Intestinal Mucosal Inflammation Through Suppression of Th1/Th17 Cell Immune Responses in Ulcerative Colitis.

Regulator of G-protein signalling (RGS) 10 plays critical roles in several immune related diseases. However, whether RGS10 is involved in colonic inflammation of ulcerative colitis (UC) is still obscure. This study aimed to investigate the role of RGS10 in UC. In this study, RGS10 expression was examined by quantitative real-time polymerase chain reaction (qRT-PCR), western blotting, immunohistochemistry, and immunofluorescent analysis. Single-cell RNA sequencing of intestinal mucosa was performed to identify key immune cells with differentially expressed RGS10. RGS10 knockout mice were generated and established dextran sulphate sodium (DSS)-induced colitis. Expression of inflammatory cytokines on mRNA and protein levels was detected by qRT-PCR, enzyme-linked immunosorbent assay, and flow cytometry. We found that RGS10 expression was significantly elevated in UC patients, especially in CD4+ T cells, compared with healthy subjects. Intriguingly, RGS10 deficiency markedly alleviated DSS-induced colitis and decreased the proportion of Th1 and Th17 cells in lamina propria mononuclear cells (LPMCs), peripheral blood (PB), spleens, and mesenteric lymph nodes (mLNs). Mechanistically, RGS10 deficiency blocked the differentiation of Th1 and Th17 cells by inhibiting the phosphorylation of signal transducer and activator of transcription (STAT) 1 and STAT3. The co-immunoprecipitation analysis further showed that RGS10 could interact with protein tyrosine phosphatase non-receptor type 2 (PTPN2), and further regulated Th1 and Th17 cells differentiation of CD4+ T cells. In conclusion, RGS10 deficiency alleviated intestinal mucosal inflammation through inhibition of Th1/Th17 cell-mediated immune responses via interaction with PTPN2 in CD4+ T cells. Therefore, targeting RGS10 may represent a novel therapeutic approach for UC treatment.

Composition of gut microbiota in obese and normal-weight Uygur adults and its association with adipocyte-related factors

Obesity is a serious global health issue. Emerging evidence indicates that the gut microbiota may contribute to the development of obesity, possibly by instigating inflammatory processes. The objective of this research is to conduct a comparative analysis of the gut microbiota composition in obese and normal-weight Uygur adults, while examining the associations with adipocyte-related factors and dietary variables. According to the inclusion and exclusion criteria, twenty-seven Uygur adults with obesity and twenty Uygur adults with normal-weight were recruited from a local community. Anthropometric measurements and blood samples were collected. Gut microbiota composition was analyzed using 16 S rRNA gene sequencing. Adipocyte-related factors were measured using enzyme-linked immunosorbent assay (ELISA). Statistical analyses were performed to compare the gut microbiota composition between the two groups and to identify correlations between gut microbiota and adipocyte-related factors. Compared with the normal-weight group, the obese group exhibited a marked reduction in both diversity and richness of the gut microbiota, alongside a decrease in Ruminococcaceae_UCG_014, Coprococcus_2, and Parabacteroides, and an increase in Megamonas and Lachnoclostridium, implying a potential link to the development of obesity. Individuals with obesity were found to have higher Leptin (LEP), Interleukin-6 (IL-6), and C-reactive protein (CRP) than normal-weight individuals. Obese Uygur adults exhibited a gut microbiota characterized by diminished diversity and richness relative to normal-weight individuals. Parabacteroides, Megamonas, and Lachnoclostridium may play an important role in the development of obesity in Uygur population. Underlying mechanisms need further investigation.

Correction of immune disorders under the influence of pharmacotherapy and extracorporeal magnetic stimulation with patients after surgical treatment of prostate adenoma

BACKGROUND: The course of benign prostatic hyperplasia is associated with immunological disorders at the systemic and local level. Despite the wide arsenal of pharmacological agents for the treatment of benign prostatic hyperplasia, some patients undergo surgical treatment. Postoperative management programmes should be aimed at reducing the risks of complications, including infectious-inflammatory complications. In this regard, the inclusion of immunomodulatory pharmacotherapy and therapeutic physical factors in the course of reconstructive treatment is reasonable and justified. AIM: To study the role of the complex effect of pharmacotherapy and extracorporeal magnetic stimulation in the correction of immune disorders with the patients after surgical treatment of benign prostatic hyperplasia. MATERIALS AND METHODS: There have been observed 88 patients after surgical treatment of benign prostatic hyperplasia, who underwent a medical rehabilitation at stages II and III. At stage II 43 patients in the comparison group underwent standardized drug therapy, individual exercise therapy; in the main group 45 patients additionally were prescribed immunomodulatory therapy with Tameritis. At stage III the patients of the comparison group continued pharmacotherapy, exercise therapy, and transrectal magnetic therapy; in the main group 45 patients were additionally prescribed extracorporeal magnetic stimulation. A method of enzyme-linked immunosorbent assay was used to study the content of interleukins (IL); IPSS-QOL (International Prostate Symptom Score – Quality-of-Life) and SF-36 (The Short Form-36) were used to assess the quality of life. Confidence level (р) 0.05. RESULTS: After the second inpatient stage of rehabilitation the patients of the main group after surgical treatment of benign prostatic hyperplasia had a decrease the level of pro-inflammatory IL by 47.5% on average (p 0.01) with an increase in the level of anti-inflammatory IL-10 by 36.2% (p 0.01). After the III outpatient rehabilitation stage, a positive dynamics significantly increased and amounted to 63.2% (p 0.01) and 51.9% (p 0.01), reaching the normative values in the population. It should be noted that the achieved results have been preserved in a long time. In the comparison group the positive dynamics was significantly lower (p 0.05; p 0.01) in all the cases after the II and III stages of rehabilitation. The patients of the main group were observed a decrease in the severity of urinary disorders (p 0.01), restoration of quality of life (p 0.05) according to IPSS-QOL and SF-36 questionnaires at significant inter-group values (p 0.05) with the same confidence. CONCLUSION: The integrated use of pharmacotherapy and extracorporeal magnetic stimulation with the patients after surgical treatment of benign prostatic hyperplasia provides significant correction of immune disorders, restoration of physical and mental health.

β-Sitosterol Mitigates Apoptosis, Oxidative Stress and Inflammatory Response by Inactivating TLR4/NF-кB Pathway in Cell Models of Diabetic Nephropathy.

Podocyte injury plays a pivotal role in the pathogenesis of diabetic nephropathy (DN), leading to proteinuria formation. β-Sitosterol is a natural compound with anti-inflammatory, anti-diabetic, nephroprotective and antioxidant properties. The studyaimed to explore whether and how β-Sitosterol protected podocytes against high glucose (HG)-induced inflammatory andoxidative injury. DN cell models were established by stimulating podocytes or renal tubular epithelial cells (HK-2) cells with 25 mM glucose. Cell viability and apoptosis were evaluated using cell counting kit-8 assays and flow cytometry analyses. Westernblotting was used to quantify protein levels of genes related to podocyte injury, HK-2 cell damage, inflammation, and TLR4/NF-кB pathway. Contents of oxidative stress biomarkers were evaluated by corresponding commercial kits while proinflammatorycytokine levels were determined by enzyme-linked immunosorbent assay. Immunofluorescence staining was performed todetect intracellular levels of reactive oxygen species (ROS) and Nrf2 nuclear translocation. Experimental results revealed that HG treatment induced podocyte dysfunction by impairing cell viability while accelerating theapoptosis, and the changes were reversed by β-sitosterol treatment. Moreover, β-sitosterol repressed HG-evoked oxidative stressby reducing ROS and malondialdehyde (MDA) levels while increasing activities of antioxidant enzymes. The reduction ofproinflammatory cytokines mediated by β-sitosterol in HG-stimulated podocytes suggested the anti-inflammatory role of β-sitosterol. Additionally, the activation of the TLR4/NF-кB signaling induced by HG was inhibited by β-sitosterol in podocytes.Inactivation of the TLR4 using TAK-242 enhanced the protective effects of β-sitosterol against HG-mediated oxidative stressand inflammation. Similarly, β-sitosterol also protected HK-2 cells from HG-induced oxidative stress, inflammation, andapoptosis. In summary, β-sitosterol exerts anti-inflammatory, anti-oxidative, and anti-apoptotic activities in HG-induced podocytes or HK-2 cells by inhibiting TLR4/NF-кB signaling.

Acylated Anthocyanins From Black Carrots and Their Related Phenolic Acids Diminish Priming and Activation of the NLRP3 Inflammasome in THP-1 Monocytes.

Excessive activation of the nucleotide-binding oligomerization domain-like receptor pyrin domain-containing protein 3 (NLRP3) inflammasome contributes to chronic inflammation. Thus, targeting NLRP3 inflammasome activation by anthocyanins may prevent inflammatory diseases. Therefore, the present study determines the influence of a black carrot extract (BCE) with high amounts of acylated anthocyanins and their related phenolic acids on the NLRP3 inflammasome. THP-1 monocytes are pretreated with a BCE, cyanidin-3-glucoside (C3G), or hydroxycinnamic acids. NLRP3 inflammasome assembly is initiated by priming THP-1 monocytes with lipopolysaccharide and/or activating the NLRP3 inflammasome with nigericin. Flow cytometry is used to assess apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) speck formation, as well as ASC and NLRP3 protein expression. Caspase-1 activity is measured using a bioluminescent assay, and cytokine concentrations are determined by enzyme-linked immunosorbent assays (ELISA). C3G and phenolic acids diminish ASC and NLRP3 protein expression. In addition, C3G and phenolic acids attenuate ASC speck formation. Furthermore, the BCE and C3G decline caspase-1 activity. Consistently, IL-1β and IL-18 secretion are reduced upon NLRP3 inflammasome activation. The present study shows that a BCE with high amounts of acylated anthocyanins and their related phenolic acids diminish priming and activation of the NLRP3 inflammasome in THP-1 monocytes.

Fecal glycoprotein 2 is a marker of gut microbiota dysbiosis and systemic inflammation

BackgroundAntimicrobial autoantigenic glycoprotein 2 (GP2) is an important component of the innate immune system which originates from the exocrine pancreas as well as from the small intestines. The relationship of GP2 with the intestinal microbiome as well as the systemic implications of increased fecal GP2 levels are, however, still unclear. Therefore, fecal samples from 2,812 individuals of the Study of Health in Pomerania (SHIP) were collected to determine GP2 levels (enzyme-linked immunosorbent assay) and gut microbiota profiles (16 S rRNA gene sequencing). These data were correlated and associated with highly standardised and comprehensive phenotypic data of the study participants.ResultsFecal GP2 levels were increased in individuals with higher body mass index and smokers, whereas lower levels were found in case of preserved exocrine pancreatic function, female sex or a healthier diet. Moreover, higher GP2 levels were associated with increased serum levels of high-sensitivity C-reactive protein, loss of gut microbial diversity and an increase of potentially detrimental bacteria (Streptococcus, Haemophilus, Clostridium XIVa, or Collinsella). At the same time, predicted microbial pathways for the biosynthesis of beneficial short-chain fatty acids or lactic acid were depleted in individuals with high fecal GP2. Of note, GP2 exhibited a stronger association to overall microbiome variation than calprotectin.ConclusionFecal GP2 is a biomarker of gut microbiota dysbiosis and associated with increased systemic inflammation. The intestines may be more important as origin for GP2 than pancreatic acinar cells. Future studies need to investigate the potential clinical value in disease specific patient cohorts.

Immunomodulatory Effects of a Prebiotic Formula with 2'-Fucosyllactose and Galacto- and Fructo-Oligosaccharides on Cyclophosphamide (CTX)-Induced Immunosuppressed BALB/c Mice via the Gut-Immune Axis.

Obejectives: This study explored the immunomodulatory effects of a prebiotic formula consisting of 2'-fucosyllactose (2'-FL), galacto-oligosaccharides (GOSs), and fructo-oligosaccharides (FOSs) (hereinafter referred to as 2FGF) in cyclophosphamide (CTX)-induced immunosuppressed BALB/c mice and its underlying mechanisms. Methods: Sixty healthy female BALB/c mice were randomly divided into the following groups: normal control (NC) group; CTX treatment (CTX) group; 2FGF low-dose (2FGF-L) group; 2FGF medium-dose (2FGF-M) group; and 2FGF high-dose (2FGF-H) group. An immunosuppressed model was established in the 2FGF-H group by intraperitoneal injection of 80 mg/kg CTX. After 30 days of 2FGF intervention, peripheral blood, spleen tissue, thymus tissue, and intestinal tissue from the mice were collected and analyzed. The changes in weight and food intake of the mice were recorded weekly. Hematoxylin-eosin (HE) staining was used to observe the histological change of the spleen tissue. Enzyme-linked immunosorbent assay (ELISA) was employed to detect cytokine levels in peripheral blood. Flow cytometry was used to analyze T lymphocyte subgroup ratio of splenic lymphocytes. Western blot analysis was conducted on intestinal tissues to assess the expression of proteins involved in the tight junction, toll-like receptor 4 (TLR4), mitogen-activated protein kinase (MAPK), and nuclear factor kappa-light-chain-enhancer of activated B cell (NF-κB) signaling pathways. Additionally, molecular techniques were used to analyze the intestinal microbiota. Results: The results showed that 2FGF restored CTX-induced splenic injury, increased the number of splenic T lymphocytes, and elevated serum cytokines such as interleukin-4 (IL-4) and IL-10. In the intestine, 2FGF upregulated the expression of intestinal epithelial tight junction proteins such as Claudin-1 and zonula occludens 1 (ZO-1), thereby enhancing intestinal barrier function and activating the MAPK and NF-κB pathways via TLR4. Furthermore, 2FGF elevated the α-diversity (Shannon and Simpson indices) of the gut microbiota in CTX-induced immunosuppressed mice, enriching bacteria species positively correlated with anti-inflammatory cytokines (e.g., IL-4) such as g_Streptomyces and g_Bacillus and negatively correlated with pro-inflammatory cytokines (e.g., IL-1β) such as g_Saccharomyces. The results suggest that 2FGF may enhance immunity via the gut-immune axis. Conclusions: The 2FGF prebiotic formula showed an immunomodulatory effect in CTX-induced immunosuppressed mice, and the mechanism of which might involve optimizing the gut flora, enhancing intestinal homeostasis, strengthening the intestinal barrier, and promoting the expression of immune factors by regulating the TLR-4/MAPK/NF-κB pathway.

Tong-Qiao-Huo-Xue Decoction promotes synaptic remodeling via cAMP/PKA/CREB pathway in vascular dementia rats

BackgroundTong-Qiao-Huo-Xue Decoction (TQHXD) is a traditional Chinese medicinal formula widely used in the treatment of vascular dementia (VD). Although it has demonstrated good clinical efficacy, the specific molecular mechanisms underlying its therapeutic effects on VD remain unclear. ObjectiveThis study aimed to elucidate the neuroprotective mechanisms of TQHXD to provide a scientific basis for the clinical treatment of VD. MethodsThe chemical components of TQHXD were qualitatively analyzed using ultra-performance liquid chromatography (UPLC) and gas chromatography (GC). Network pharmacology predicted the potential protective mechanisms of TQHXD against VD. A rat model of VD was established through bilateral vessel occlusion (2-VO), and an oxygen-glucose deprivation/reperfusion (OGD/R) model was used to induce damage to neuronal cells of the hippocampus. In vivo experiments assessed changes in cerebral blood flow, learning and memory capabilities, hippocampal neuronal morphology, dendritic length, dendritic spine density, and synapse number in rats. We examined the expression of synaptic remodeling-related proteins and pathway proteins in the hippocampal region. In vitro assays evaluated cell viability, apoptosis, reactive oxygen species (ROS) levels, and expression of synaptic remodeling-related proteins and signaling pathway. ResultsMultiple active components were identified in TQHXD. KEGG enrichment analysis suggested that the therapeutic effects of TQHXD on VD may be related to the cAMP signaling pathway. Treatment with TQHXD significantly improved learning and memory performance in VD rats, improved hippocampus morphology, and increased dendritic length, dendritic spine density, and number of synapses. Furthermore, TQHXD improved cell viability, reduced apoptosis, and decreased intracellular ROS levels in vitro. Western blotting, immunofluorescence, and enzyme-linked immunosorbent assay results collectively demonstrated that TQHXD upregulated the expression of synaptic remodeling-related proteins and pathway-related proteins both in vivo and in vitro. ConclusionsTQHXD treated VD by promoting synaptic remodeling in hippocampal neurons, likely through activation of the cAMP/PKA/CREB pathway.

Key predictors of fertility: Exploring the role of Vitamin-D

Background &amp; Objectives: The relationship of Vitamin-D (VD) with Luteinizing Hormone (LH), Follicle stimulating Hormone (FSH) and Anti Mullerian Hormone (AMH) is recognized. This study was conducted to determine whether VD is a predictor of fertility and investigate its relationship with female reproductive hormones and markers of ovarian reserve in female population of Karachi. Methods: A cross-sectional study was performed from July 2020 to June 2022 at the Aga Khan University Hospital (AKUH) with recruitment of 135 fertile and infertile subjects. VD, AMH, FSH and LH levels were estimated by ‘Enzyme Linked Immunosorbent assay (ELISA)’. By applying binary logistic regression, variables with p-value &lt; 0.25 in univariate analysis were used for multivariate regression model and adjusted odds ratios were computed. In multivariate analysis, significance was p-value ≤ 0.05. Results: VD deficiency was observed in all infertile female subjects. In univariate regression analysis, FSH, LH, LH/FSH ratio and VD levels turned out to be significant. The Multivariate analysis represented that for every one ng/mL increase in VD level, the odds of fertility were expected to be 50.154 times higher with p-value &lt; 0.001. The positive correlation of VD with FSH was significant. Conclusion: VD deficiency was prevalent in all infertile female subjects. VD levels were strong predictor of fertility in the study population. The significant association of VD with FSH explained the indirect effect of VD on follicular development and ovarian functions however no relationship of VD with ovarian reserve was exhibited. Note: AI assisted technologies were used in the preparation of the manuscripts, to improve readability and language. doi: https://doi.org/10.12669/pjms.40.10.9774 How to cite this: Salahuddin H, Zaki S, Ashraf M, Rehana Rehman4. Key predictors of fertility: Exploring the role of Vitamin-D. Pak J Med Sci. 2024;40(10):2363-2367. doi: https://doi.org/10.12669/pjms.40.10.9774 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Analysis of Mannose-Binding Lectin Protein and mRNA Levels on Selected Chicken Breeds in South Africa.

Mannose-binding lectin (MBL) is a key component of the innate immune system that plays a crucial role in binding to the microbial sugar surface to recognize and eliminate pathogens by activating the complement system. To detect and quantify the MBL protein concentration and chicken MBL expression in selected chicken breeds in South Africa. Forty-five blood samples from three indigenous chicken breeds, Ovambo (OV=9), Venda (VD=9) and Potchefstroom Koekoek (PK=9), and two exotic chicken breeds, Rhode Island Red (RIR=9) and Lohmann Brown (LB=9), were used for MBL protein concentration using enzyme-linked immunosorbent assay (ELISA) techniques. Also 20 liver samples from symptomatic two indigenous chicken breeds, OV (5) and PK (5), and two exotic chicken breeds, RIR (5) and LB (5), were used for MBL expression using quantitative polymerase chain reaction (qPCR) techniques. A general linear model was done using Tukey's multiple comparison post hoc test. The findings revealed MBL protein concentration from 5.26 to 18.56µg/mL. The LB breed had the lowest mean 6.40±0.80µg/mL, whereas the PK breed had the highest mean MBL concentration of 17.70±0.24µg/mL of MBL protein concentration. At 12, 25 and 35 weeks, the MBL proteins of OV, VD, PK, RIR and LB varied significantly at p≤0.05. The mRNA MBL expression of OV and LB breeds showed a 1-fold decrease in MBL expression, while RIR showed a 2-fold increase in MBL expression, and the PK showed more than a 3-fold increase in MBL expression relative to the control. The least-squares means for OV, LB, PK and RIR mRNA MBL expression were 0.54±0.19, 0.68±0.30, 4.46±2.76 and 2.89±0.19µg/mL, respectively. MBL protein was detected and quantified with distinct differences in concentration and expression levels at the presence of mycoplasma gallisepticum among the sampled South African chicken breeds. This highlights the genetic diversity of MBL as a tool for disease prevention in South African chicken breeds.

Cross-reactivity between dengue virus and SARS-CoV-2 antibodies: Confirmation study using specimens from dengue-infected patients before the COVID-19 pandemic

BackgroundThe simultaneous occurrence of the COVID-19 pandemic and a dengue outbreak has posed significant challenges for governments and medical personnel in dengue-endemic countries like Indonesia. Several studies in dengue-endemic countries have reported cases of misdiagnosis between COVID-19 and dengue. Therefore, it is crucial to evaluate the potential cross-reactivity between SARS-CoV-2 antibodies and dengue. MethodsThis study aimed to confirm the serological cross-reaction between dengue virus and SARS-CoV-2 in Surabaya, East Java, which is a highly dengue-endemic city in Indonesia. In total, 238 serum samples with confirmed dengue that were collected before the emergence of COVID-19 were tested to detect the presence of reacting IgG and IgM antibodies (Abs) against SARS-CoV-2 via a rapid detection test (RDT) and enzyme-linked immunosorbent assay (ELISA). Samples from patients with dengue infection collected during the pandemic, from healthy volunteers predating the pandemic, and from patients with COVID-19 were used for comparison. Results and conclusionFew (6.7 %) of the pre-COVID-19 dengue Ab-positive serum samples showed reactive on SARS-CoV-2 in the RDT, with significantly lower IgG and IgM levels detected in ELISA compared with the dengue samples collected during the pandemic and the COVID-19 samples (P < 0.005). A comparable anti-SARS-CoV-2 IgG concentration was observed in the pre-COVID-19 dengue samples and healthy volunteers (P = 0.56), which also indicated other possibilities. In conclusion, our results suggested a low risk of cross-reactivity between dengue virus and SARS-CoV-2. However, they highlighted the need for caution when using and interpreting data obtained stemming from serological methods, to prevent false-positive results. Further studies are needed to evaluate the cross-reactivity between dengue virus, SARS-CoV-2, and other common human pathogens, as well as its effect on the serosurveys, treatment of these diseases, or vaccine efficacy.

Vascular endothelial growth factor/connective tissue growth factor and proteomic analysis of aqueous humor after intravitreal conbercept for proliferative diabetes retinopathy.

To investigate the role of connective tissue growth factor (CTGF) and vascular endothelial growth factor (VEGF) in the protein profile of the aqueous humor in patients with proliferative diabetic retinopathy (PDR) following intravitreal injection of conbercept. This study included 72 PDR patients and 8 cataract patients as controls. PDR patients were divided into 3 groups according to the intervals of 3, 5, and 7d between intravitreal conbercept (IVC, 0.5 mg/0.05 mL) injection and pars plana vitrectomy (PPV) performed. Aqueous humor samples were collected before and after IVC and PPV for VEGF and CTGF levels detected with enzyme-linked immunosorbent assay (ELISA). The differential proteomics of 10 patients who underwent PPV surgery 5d after IVC and 8 normal controls was studied, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were performed on the data, and the protein interaction network of 23 differential proteins was studied. Post-IVC, VEGF levels decreased and CTGF levels increased significantly in aqueous humor, with the CTGF/VEGF ratio rising significantly at all intervals. Liquid chromatography tandem mass spectrometry (LC-MS/MS) identified differentially expressed proteins between pre- and post-IVC samples. GO and KEGG analyses revealed involvement in immune response, stress response, complement and coagulation cascades, ferroptosis, and PPAR signaling pathways. PPI analysis highlighted key proteins like APOA1, C3, and transferrin (TF). ELISA assay confirmed the differential expression of proteins such as HBA1, SERPINA1, COL1A1, and ACTB, with significant changes in the IVC groups. The study demonstrates that IVC effectively reduces VEGF levels while increasing CTGF levels, thereby modifying the CTGF/VEGF ratio, and IVC significantly alters the protein profile in the aqueous humor of patients with PDR. Proteomic analysis reveals that these changes are associated with critical biological pathways and protein interactions involved in immune response, stress response, and cellular metabolism.

Systemic profile of pro-inflammatory and anti-inflammatory cytokines and its changes in patients with cataract after surgery

Background. Due to its high prevalence and morbidity, cataract is an urgent medical and social problem. In recent years, immune inflammation, in particular the immune regulation of dystrophic processes, has been of great importance in the pathogenesis of eye diseases. Immunological disorders play a crucial role in the development of age-related eye diseases (cataract, corneal dystrophy, glaucoma, age-related macular degeneration). The effect of pro-inflammatory and anti-inflammatory cytokines on the development of age-related cataract remains understudied, as well at their role in predicting the course of the postoperative period after extraction of age-related cataract has not been fully stu­died, which determines the relevance of the study. The objective: to assess the state of cytokine profile in patients with age-related cataract by determining the content of pro-inflammatory (IL-1β, IL-6) and anti-inflammatory (IL-4, IL-10) interleukins in blood serum before and after surgical treatment of cataract. Materials and methods. The level of pro-inflammatory (IL-1β, IL-6) and anti-inflammatory cytokines (IL-4, IL-10) was studied before and after surgical treatment in 28 patients with age-related cataract (56 eyes), 8 men and 20 women aged 42 to 82 years. The average age of the patients was 71.79 ± 1.90 years. Serum content of cytokines was determined by the enzyme-linked immunosorbent assay using test systems Vector-Best-Ukraine. Results. There was no excess of IL-1β and IL-6 reference values in the blood serum of patients with age-related cataract prior to surgical treatment. Analysis of the pro-inflammatory IL-6 in these patients depending on sex revealed a significant increase in its level by 1.3 times in men and a significant decrease by 1.1 times in women compared to its average content (p &lt; 0.06 and p &lt; 0.01, respectively). The level of anti-inflammatory IL-4 and IL-10 in the blood serum of the examined patients was within the reference values. When studying the content of pro-inflammatory and anti-inflammatory interleukins in serum depending on sex and age in patients with age-related cataract aged 75 years and older, there was a statistically significant increase in IL-6 and IL-10. In this age group, the level of IL-6 was 1.4 times higher in men compared to women, and IL-10 was 1.8 times higher in women than in men. In patients with age-related cataract after surgical treatment, serum content of pro-inflammatory IL-1β, IL-6 and anti-inflammatory IL-4, IL-10 was higher than levels before surgery. The highest levels of IL-1β, IL-6 were in patients with postoperative corneal edema. Serum content of IL-6 in patients with postoperative corneal edema was higher compared to that of IL-1β. Levels of IL-4, IL-10 were higher in patients with postoperative corneal edema, IL-10 in these patients prevailed over IL-4. Conclusions. In patients with age-rela­ted cataract, the content of pro-inflammatory and anti-inflammatory cytokines in the blood serum corresponds to reference values and has no signi­ficant differences from the control group. However, determination of the cytokine profile in people with age-related cataract revealed a violation of the systemic immunological status depending on age and gender. According to our results, patients with age-related cataract over 75 years of age had a significant increase in the level of pro-inflammatory cytokine IL-6 and anti-inflammatory cytokine IL-10 in blood serum. In this age group, IL-6 was higher in men and IL-10 was higher in women. We also noted the absence of significant differences in the content of IL-1β, which may indicate a more specific value of IL-6 levels for the development of cataract. In patients with age-related cataract after surgical treatment, serum levels of pro-inflammatory IL-1β, IL-6 and anti-inflammatory IL-4, IL-10 were higher than preoperative ones. The highest levels of IL-1β, IL-6 were in the blood serum of patients with postope­rative corneal edema, and a decrease in the level of IL-4, IL-10 was also noted in cases of corneal edema. Serum content of IL-6 in patients with postoperative corneal edema was higher than that of IL-1β, and IL-10 prevailed over IL-4 levels.

Crimean Congo hemorrhagic fever virus exposure among febrile patients, cattle herders, and cattle herds in Cameroon

Problem addressedCrimean Congo hemorrhagic fever (CCHF) is a tick-borne disease with high fatality rates and an expansive geographic distribution, yet disease prevalence data in Cameroon is lacking. ObjectiveThis study aimed to determine CCHF virus (CCHFV) seroprevalence and tick distribution among cattle herders and febrile patients in West and Centre Cameroon. Methods and approachTwo cross-sectional serological studies of human and cattle were conducted from October to December 2021 and from June to July 2022, which included the collection of ticks. Enzyme-linked immunosorbent assay (ELISA) were used to detect anti-CCHFV antibodies, while a knowledge, attitudes, and practice (KAP) survey assessed tick and tickborne disease related knowledge and behaviors among herders. Tick identification used morphological keys. ResultsThe KAP survey showed adequate tick knowledge (94.5 %) among herders but poor understanding of disease transmission, with favorable attitudes towards tick control (24.7 %) but inadequate implementation. Rhipicephalus annulatus (64.1 %) predominated among the 1,296 ticks collected during each rainy season. Among cattle, 27.4 % were seropositive, and seropositivity was associated with specific villages, cattle age (>4 years), and female sex. Herders had a 17.8 % seroprevalence, while febrile patients had 8.3 %, with higher rates in those >20 years old for both groups. Self-reported tick removal by herders after contact and grazing may increase CCHFV exposure. ConclusionsThis study confirms CCHFV circulation in rural West Cameroon and unexpected exposure risk in Yaounde, highlighting the need for active entomological surveillance and preventive measures in transhumance and cattle market activities. Establishing an occupation-based surveillance system can help identify CCHFV hotspots to prevent outbreaks.

Comparison of orexigenic and anorexigenic neuropeptide levels in hyperemesis gravidarum patients with normal pregnant women: A prospective cohort study.

The aim of this study was to determine whether orexigenic neuropeptides, orexin and galanin, and anorexigenic neuropeptides, alpha-melanocyte-stimulating hormone (α-MSH) and cocaine- and amphetamine-regulated transcript (CART), are implicated in hyperemesis gravidarum (HG). Fifty pregnant women who had been diagnosed with HG between April 2022 and February 2023 at the Siirt University Faculty of Medicine Training and Research Hospital (tertiary center) were recruited for this study. An equal number of pregnant women without an HG diagnosis were included in the study as the control group. Participants' age, pregnancy history, medical history, thyroid function test results, complete blood count results, and electrolyte levels were recorded, and their orexin, galanin, α-MSH, and CART serum levels were analyzed using an enzyme-linked immunosorbent assay. No statistically significant differences in orexigenic neuropeptides (orexin and galanin) were observed between the HG and control groups. A statistical difference was found between an anorexigenic neuropeptide (α-MSH) and the control group (P = .012). Based on a receiver operating characteristic analysis, the α-MSH parameter was statistically significant for distinguishing between participants with an HG diagnosis and those without, with a sensitivity of 63.6%, specificity of 65.9%, and cutoff value of 11769.3 pg/mL (P = .012, area under curve: 0.655). Based on the severity classification of ketonuria (ketonuria levels of +1 or +2 were classified as mild, whereas levels of +3 or +4 were classified as moderate to severe), the anorexigenic CART neuropeptide was found to be a statistically significant diagnostic indicator of severe ketonuria (P = .020). α-MSH and CART levels were found to be related in HG patients and in HG patients with severe ketonuria.