ELISA Technique Research Articles

Serum High Mobility Group Box 1 (HMGB1) Protein Levels and Cognitive Function in Epilepsy Patients: A Cross-Sectional Study

Background: Epilepsy is a neurological disease with a high incidence rate. Cognitive decline is one of the consequences of recurrent seizures. Neuroinflammation is closely related to the development of epilepsy and cognitive impairment. An increase in the expression and translocation of High Mobility Group Box 1 (HMGB1) from the nucleus to the extracellular space has been observed in epilepsy patients and experimental animal models. This study aimed to investigate the relationship between serum HMGB1 levels and cognitive function in epilepsy patients. Methods: This cross-sectional observational study involved 45 epilepsy patients. Cognitive function was assessed using the Indonesian version of the Montreal Cognitive Assessment (MoCA-Ina), and serum HMGB1 levels were measured using the ELISA technique. The relationship between cognitive function and HMGB1 levels was analyzed using the Kruskal-Wallis test, with a significance level set at p < 0.05. Results: The mean age of the participants was 28.5 years, with a higher proportion of females. The mean serum HMGB1 level was 22.6 ng/ml. No significant relationship was found between serum HMGB1 levels and cognitive function in epilepsy patients (p = 0.188). Conclusion: Serum HMGB1 protein levels were not associated with cognitive function in this sample of epilepsy patients.

Increased serum EGF but not SDF-1 levels are associated with the pathophysiology and development of generalized anxiety disorder

Generalized anxiety disorder (GAD) is a common and persistent mental illness accompanied by uncontrollable worries for daily staff. Physiological, environmental, genetic, and daily stress from surroundings is involved in developing GAD. Here, we aimed to assess the association of stromal cell-derived factor-1 (SDF-1) and epidermal growth factor (EGF) in the pathophysiology of GAD and their role as diagnostic tools. This case-control study recruited 50 GAD patients from the psychiatry department of a tertiary care teaching hospital in Dhaka city and 38 age-sex-matched healthy controls (HCs) from the surrounding areas. A qualified psychiatrist conducted standardized psychiatric interview to diagnose GAD patients and evaluate HCs by applying the diagnostic and statistical manual for mental disorders, 5th edition (DSM-5) and used the GAD-7 scale to assess the severity of the GAD symptoms. After confirming the inclusion and exclusion criteria, we collected 5 ml blood samples from each participant. We measured serum SDF-1 and EGF levels using ELISA techniques. We observed increased serum levels of EGF in GAD patients compared to HCs (5.91 ± 3.90 ng/ml vs. 1.59 ± 1.08 ng/ml; p < 0.001). Also, this increment is positively associated with the severity of GAD in patients (r = 0.479, p = 0.002). The receiver operating characteristic analysis showed good diagnostic performance (AUC 0.869, p < 0.001) with high sensitivity (82.5%), and specificity (81.1%) at cut off value of 2.31 ng/ml. However, we didn’t find significant differences in serum SDF-1 levels between GAD patients and HCs (11.96 ± 4.50 ng/ml vs. 12.70 ± 11.76 ng/ml; p = 0.337). The present study suggests that GAD patients have increased serum EGF levels but not SDF-1 levels compared to HCs and that increased EGF levels are associated with increased GAD symptoms as well. Moreover, EGF levels could serve as a biomarker for GAD However, further interventional studies with larger and homogeneous samples are suggested to confirm and establish these findings.

Petunidin attenuates vinclozolin instigated testicular toxicity in albino rats via regulating TLR4/MyD88/TRAF6 and Nrf-2/Keap-1 pathway: A pharmacodynamic and molecular simulation approach

Vinclozolin (VZN) is a widely used fungicide which exerts deleterious impacts on various organs including testis. Petunidin (PDN) is a polyphenolic compound that demonstrates a broad range of pharmacological activities. Thirty-two rats were divided into 4 groups including the control, VZN (100 mg/kg), VZN (100 mg/kg) + PDN (4 mg/kg) and PDN (4 mg/kg) treated group. The activities of antioxidant enzymes were assessed by using previously documented protocols. The gene expressions were determined by using qRT-PCR. The levels of hepatic function and apoptotic markers were evaluated by using standard ELISA technique. The histological analysis was carried out as per the standard protocol of histology. It was revealed that VZN disrupted the Nrf-2/Keap-1 pathway. Moreover, the activities of catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), heme-oxygenase-1 (HO-1) and glutathione reductase (GSR) were reduced whereas levels of reactive oxygen species (ROS) & malondialdehyde (MDA) were promoted following the VZN intoxication. Furthermore, VZN intoxication reduced total sperm count, viability, motility as well as luteinizing hormone (LH), follicle stimulating hormone (FSH), and plasma testosterone. Besides, administration of VZN decreased the expressions of 3β-Hydroxysteroid dehydrogenase (3β-HSD), steroidogenic acute regulatory protein (StAR) and 17β-Hydroxysteroid dehydrogenase (17β-HSD). Moreover, VZN exposure escalated the expressions of Bcl-2–associated X protein (Bax) and cysteine–aspartic acid protease-3 (Caspase-3) while reducing the expressions of B-cell lymphoma-2 (Bcl-2). Additionally, VZN administration increased the gene expression of toll-like receptor 4 (TLR4), tumor necrosis factor receptor-associated factor 6 (TRAF-6) and myeloid differentiation primary response 88 (MyD88). The levels of interleukin-6 (IL-6), nuclear factor kappa-B (NF-κB), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and the activity of cyclooxygenase-2 (COX-2) were promoted following the VZN administration. Furthermore, VZN intoxication disrupted the normal histology of testicular tissues. However, VZN + PDN treatment ameliorated testicular damage via regulating aforementioned dysregulations owing to its anti-inflammatory, antioxidative as well as anti-apoptotic potentials. Lastly, molecular docking (MD) was performed to assess the effectiveness of PDN as a curative compound by analyzing its binding affinity with the targeted proteins (Keap1, TLR4 and StAR). Our in-silico evaluations confirmed that PDN possesses the potential to interact with binding pockets of these proteins, emphasizing its capability as a curative compound to mitigate VZN-prompted reproductive damage.

Hepatitis B virus infection among hemodialysis patients in Aden-Yemen

HBV infection is one of major global health problems worldwide. It increases the morbidity and mortality rates among hemodialysis patients which make the management process more complicated. This study was aimed to determine the prevalence of HBV infection among hemodialysis patients and assessment the risk factors associated with infection. An analytical cross-sectional study was conducted on 50 HD patients. Blood samples were collected and serum HBsAg was analyzed using ELISA technique. The overall prevalence of HBV was 6 (12.0%). The males had highest rate. There were statistically significant recorded between the prevalence of HBV infection and age group 30 – 39 years and duration of hemodialysis from 1-2 years and more than 2 years (p=0.02, 0.001 and 0.001) respectively. It can be concluded that the HBV prevalent rate among HD patients was common and lower than that reported among most general population in Yemen. Increasing of the duration and long standing in hemodialysis unit may increase the risk of HBV infection. Hepatitis B vaccine should be given to all HBV negative HD patients before HD process and health education programs about the transmission routes of HBV and preventive measures should be implemented for general population.

SARM1 Deficiency Induced Depressive-like Behavior via AMPKα/p-eEF2 axis to Synapse Dysfunction

Sterile Alpha and TIR Motif Containing 1 (SARM1) are proteins implicated in various neurological processes; however, their role in depression remains unexplored. This study investigated the contribution of SARM1 to depressive-like behaviors in a chronic stress-induced depression model and SARM1 knockout (KO) mice. Depressive-like behaviors were assessed using a battery of behavioral tests, including the Open Field Test (OFT), the Forced Swim Test (FST), the Sucrose Preference Test (SPT), and the Tail Suspension Test (TST). Mitochondrial energy metabolism alteration, cytokine level changes, and other related molecular signalling protein expression were evaluated using ELISA and western blotting techniques to investigate the underlying mechanisms. Behavioral assessments (OFT, FST, SPT, TST) revealed depressive-like phenotypes in SARM1 KO mice, accompanied by altered mitochondrial energy metabolism (NAD+, ATP) in the cortex. Intriguingly, SARM1 depletion led to peripheral inflammation, as evidenced by elevated cytokine levels in plasma but not in brain regions (cortex). In addition, we found dysregulated energy metabolism, AMPK signaling, and synaptic plasticity in the cortex of SARM1 KO mice. Notably, AICAR (Acadesine), an AMPK activator, ameliorated depressive-like behaviors and synaptic dysfunction, while Compound C, an AMPK inhibitor, reversed these effects. Additionally, NH125, an eEF2 kinase inhibitor, improved depressive-like behaviors in SARM1 KO mice. These findings demonstrate that SARM1 is critical in regulating depressive-like behaviours through the AMPKα/p-eEF2 signalling pathway. Targeting AMPK signaling and synaptic function may offer novel therapeutic avenues for depression.

Association Between Prostatitis and Some Biochemical Markers in Patients of the AL-Najaf Province, Iraq

One of the most common clinical conditions in men is prostate disease, especially prostatitis, which causes various symptoms, that negatively affect patient life. Our current study investigated the relationship between prostatitis and IL-6, zinc, and ESR levels for patients who visited AL-Sader Medical City in the province of AL-Najaf, between March and November 2023. our study included 90 samples of people infected with prostatitis and 30 samples of healthy people between the ages of (20-40) years. The level of IL-6 was calculated using the ELISA technique, The level of zinc as an antioxidant asses by using the spectrophotometer and ESR was measured by using the Westergren technique to determine the inflammation case in patients and compared with the healthy group. The results showed an increase in the level of IL-6 in patients with prostatitis compared with a healthy group (908.72±140.19,214.16±85.56) pg/ml respectively. but showed a decrease in the level of Zinc in prostatitis patients compared with the healthy group (38.93±10.92U/ml, 89.36±9.61U/ml) respectively. Also showed an elevated level of ESR, where according to (48.41±13.49 ) mm/hour in patients group compared with a healthy group (14.60±4.14) mm/ hour. The data was analyzed by SPSS program version 23, where the factor effect was considered significant when p&lt;0.05. we conclude from our current study the significant relationship between IL-6, Zinc levels, and prostatitis, as well as the role of the ESR test as an indicator of prostatitis

Yellow Mustard Protein a Immunoreactivity Reduction Through Seed Germination, Lactic Acid Fermentation, and Cooking

Food allergens are becoming increasingly threatening and are disrupting the health and social structure of a significantly large population worldwide. Proteins from mustard are among the well-recognized food allergens which affect many sensitive individuals. Many processing methods are continually being explored to reduce allergen immunoreactivity and for developing hypoallergenic foods. Cooking, germination, and fermentation have been evaluated to attenuate the immunoreactivity of food allergens. The objective of this study is to evaluate the effect of seed germination, lactic acid fermentation, and/or cooking on yellow mustard seed protein immunoreactivity (IR) (protein A) using ELISA techniques. Samples from five-day germination at 35–40 °C and three-day fermentation between 25 °C and 35 °C were evaluated. The germination and fermentation processes yielded varying reductions in the IRs of mustard proteins, with a combined yield of about 90% reduction. When complemented with further stovetop cooking, protein IR reduction was extended up to 98%, while cooking alone resulted only in about a 70% reduction. FTIR results confirmed that changes in mustard protein conformation maybe due to the unfolding and/or denaturation of mustard proteins. These processing methods are beneficial as they not only help reduce the native mustard protein IR, but also increased inherent antioxidant activities in germinated and fermented mustard seeds.

HIV protein profile characteristics in patients with first-time detected infection

The HIV-infection continues to be one of the most large-scale epidemics worldwide. Many techniques have been developed to detect this disease, but the Western blot based on the identification of specific viral proteins remains the most commonly used method that allows to monitor ongoing viral processes. Despite discussions regarding the criteria for a positive test assessment and selection of a minimum number of viral proteins to reliably interpret the data, a very few studies on the protein profiles in HIV-infected patients, particularly in the Russian Federation are available. The aim of this study was to assess the prevalence of HIV viral proteins in a group of people with newly diagnosed infections analyzing 2566 blood samples from individuals with newly diagnosed HIV infection for reference testing. The samples were assessed using ELISA and IHL techniques, followed by western blotting. Subsequently, the following viral proteins were analyzed to assess HIV life cycle and the predominance of its different stages: gp160, gp120, gp41, p55, p40, p24, p17, p66, p51, and p31. For comparison, gp110/120 was chosen as the reference protein due to its lowest prevalence frequency among allenvgene products comprising 96.06%. A significantly reduced prevalence frequency was found for several protein groups: GAG — p55 (80.91%), p40 (72.14%), nucleocapsid p18/17 (67.37%); POL proteins — p68/66 (89.57%), p52/51 (81.91%), p34/31 (86.02%). Significant differences in frequency of viral proteins between age and sex groups are shown. Hypotheses explaining the obtained data are presented. By aligning anti-viral protein antibody profile with the course of the infection and patient’s condition, it will be possible to identify patterns and take necessary measures for early diagnostics with extended results, such as duration of the infection, viral load, and disease severity.

N-Acetyl Cysteine and Zinc Sulfate Attenuates Acute Crude Oil-Induced Oxidative Stress and Testicular Structural Damage in Male Wistar Rats

Introduction: Crude oil toxicity is caused by the production of reactive oxygen species (ROS), leading to oxidative stress and tissue damage. Both natural and synthetic antioxidants have shown potential in reducing the impact of this stress. This study evaluated the protective effects of N-Acetyl Cysteine (NAC) and Zinc Sulfate (ZnSO4) in alleviating testicular cytoarchitectural damage and oxidative stress in male Wistar rats after short-term exposure to Bonny light crude oil (BLCO) Materials and Methods: Fifty male Wistar rats (180–200g) were divided into ten groups of five. Group I (control) received distilled water, and Group II (negative control) was given BLCO (600mg/kg). Groups III and IV were administered NAC at doses of 100mg/kg and 200mg/kg, respectively. Groups V and VI received BLCO (600mg/kg) along with 100mg/kg and 200mg/kg NAC. Groups VII and VIII were treated with ZnSO4 at doses of 0.5mg/kg and 1mg/kg, respectively, while Groups IX and X were given BLCO (600mg/kg) combined with 0.5mg/kg and 1mg/kg ZnSO4. All treatments were administered orally via an orogastric cannula for three weeks. Testicular tissue was assessed by using histopathological examinations. Also, tissue malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH) and total antioxidant capacity (TAC) were evaluated by using the ELISA technique. Results: Exposure to BLCO resulted in a significant increase in MDA and GSH levels and a significant decrease in SOD, CAT and TAC levels in BLO-exposed animals compared to control (P&lt;0.05) which were mitigated by co-treatment with NAC and ZnSO₄. Histological analysis of the testes from rats exposed to BLCO revealed notable alterations in testicular morphology, including interstitial exudate, and atrophic and deteriorated seminiferous tubules which indicate testicular degeneration. However, this was improved on supplementation with NAC and ZnSO₄. Conclusion: This study demonstrates that BLCO-exposed rats experienced toxicity affecting both testicular antioxidants and testicular structure. Our findings suggest that NAC and ZnSO4 protected and improved testicular function in BLCO-exposed rats by enhancing testicular antioxidant levels and cytoarchitecture.

4D-DIA-based proteomics analysis reveals the protective effects of Pidanjiangtang granules in IGT rat model

Ethnopharmacological relevanceThe Pidanjiangtang (PDJT) formula was founded on the "Pidan" theory from the "Nei Jing." PDJT is considered to eliminate the accumulation of pathological products, remove heat sources, and prevent damage to organs such as the liver and islets. It is widely used in clinical practice to treat impaired glucose tolerance (IGT). However, the bioactive ingredients and underlying mechanisms are still unclear and need further investigation. ObjectiveThis study aimed to determine the therapeutic effect of PDJT on IGT rats and explore the mechanism of PDJT intervention on IGT by four-dimensional independent data acquisition (4D-DIA) proteomics analysis. Materials and methodsThe IGT model was established by a high-fat diet combined with Streptozotocin (STZ) injection. The IGT rats were treated with low, medium, and high doses of PDJT orally for 42 days and compared with the Metformin positive control group. The therapeutic effects of PDJT on IGT rats were evaluated using the oral glucose tolerance test (OGTT), serum lipoprotein detection, insulin detection, liver histopathology, and hepatic steatosis assessment. 4D-DIA proteomics analysis was used to explore the differential proteins (DEPs) and potential pathways of PDJT. Finally, Western blotting and ELISA techniques were used to verify DEPs and major targets. ResultsPDJT can enhance glucose metabolism, restore islet β cell function, regulate lipoprotein metabolism, reduce hepatic steatosis, and consequently slow down the progression of IGT. In the proteomic analysis, a total of 355 DEPs were identified, and critical proteins were validated. The results indicated that the JAK2/STAT1 signaling pathway plays a pivotal role in the effects of PDJT. IκB-ζ may be a potential target for PDJT in regulating the inflammatory response of IGT. ConclusionPDJT is an effective formula for improving IGT, with its potential mechanism linked to the JAK2/STAT1/IκB-ζ signaling pathway. This study offers a novel approach to investigating the mechanisms of TCM formula through proteomics and offers new insight into exploring TCM treatment for IGT.

The Constituents Analysis and the Anti-inflammatory Activity of the Chinese Medicine: Kanglao Concoction

Introduction: The Chinese medicine, Kanglao concoction has been used clinically to treat tuberculosis. However, its constituents and anti-inflammatory activity have not been fully understood. Method: In this study, HPLC-MS2 has been employed to determine the main constituents in the concoction. The anti-inflammatory activity has been investigated by ELISA and Western Blot techniques. The MS analysis revealed that tigogenin and afzelin were two main constituents in Kanglao concoction. Result: Moreover, the result of ELISA experiments showed that Kanglao concoction significantly reduced the expression of TNF-α and IL-1β at 5 mg/mL and 10 mg/mL to inhibit inflammation. Conclusion: The expression of Notch pathway-related proteins in A549 cells increased under the stimulation of Mycobacterium tuberculosis, and decreased dose-dependently upon the treatment of Kanglao concoction, which was deduced to inhibit Notch pathway expression to impose antiinflammatory activity.

Haemolysis and acute tubular injury after pulsed field ablation compared to radiofrequency: An observational study

Abstract Background Pulsed field ablation (PFA) represents a novel promising non-thermal alternative in non-pharmacological treatment of atrial fibrillation (AF). However, infrequent cases of acute renal failure secondary to intravascular haemolysis after a PFA procedure have been described. Objective To investigate the impact of ablation technology (PFA vs. radiofrequency ablation (RFA)) on the level of plasma cell-free haemoglobin (CFH) and the concentration of serum neutrophil gelatinase-associated lipocalin (NGAL) as a marker of tubular injury. Methods This was a prospective observational trial. In a consecutive cohort of patients who underwent AF ablation (PFA of RFA), blood samples were drawn just before the procedure (Sample 1: CFH and NGAL), immediately after the procedure (Sample 2: CFH) and one day after the procedure (Sample 3: CFH and NGAL). Plasma CHF level and serum NGAL concentration were analysed using the ELISA technique. Results Among 70 patients enrolled (mean age 64.3 ± 10.3 years, 61 % male), 23 underwent RFA and 47 PFA (22 pulmonary vein isolation (PVI) only and 25 PVI with other additive lesions). Baseline serum creatinine levels of both groups were comparable (91.7 ± 22.1 µmol/L vs. 88.8 ± 22.1 µmol/L P = 0.44). In the PFA cohort, a significant peak of CFH concentration was observed just after the procedure with a subsequent decrease within 12 hours (Sample 1 vs. Sample 2: 93.4 ± 65.1 µg/mL vs. 2394.9 ± 1966.1 µg/mL P &amp;lt; 0.001, Sample 1 vs. Sample 3: 93.4 ± 65.1 µg/mL vs. 97.2 ± 68.5 µg/mL P = 0.75). No significant differences were observed in the RFA cohort. Compared to baseline, neither the PFA nor the RFA group showed a significant difference in NGAL levels (PFA 98.6 ± 31.7 ng/mL vs. 98.5 ± 38.1 ng/mL P = 0.78, RFA 108.3 ± 33.8 ng/mL vs. 116.3 ± 32.2 ng/mL P = 0.49). Conclusions Compared to RFA, PFA leads to significant peri-procedural haemolysis. However, no differences in the concentration of the tubular injury marker NGAL were observed on the first day after the procedure.

Circulating levels of neutrophil extracellular traps reflects local neutrophil activity but not fibrinolytic activity in coronary thrombi from patients with ST-elevation myocardial infarction

Abstract Introduction Neutrophil extracellular traps (NETs) are associated with impaired fibrinolysis in ischemic stroke, while NETs degradation enhances lysis of coronary thrombi in ex vivo experiments. Whether circulating NET markers are associated with local fibrinolytic activity in patients with ST-elevation myocardial infarction (STEMI) has not been reported. Purpose We investigated associations between circulating NET markers and gene expression of fibrinolytic markers in aspirated coronary thrombi from patients with STEMI. Furthermore, we explored whether local upregulation of NET formation by peptidylarginine deiminase 4 (PAD4) was reflected in the circulation. Methods Coronary thrombi from 35 STEMI patients undergoing primary percutaneous coronary intervention (PCI) were aspirated and promptly snap-frozen to -80°C. Peripheral blood samples were collected simultaneously. Median time from start of symptoms to PCI was 152 min. Thrombus gene expression of tissue Plasminogen Activator (tPA), urinary-type Plasminogen Activator (uPA), plasminogen-activation inhibitor type 1 (PAI-1) and PAD4 were quantified with RT-PCR. Gene expression of PAD4 was also measured in circulating leukocytes. Circulating NET markers were measured by a fluorescent nucleic acid stain using fluorometry (dsDNA), an in-house ELISA technique (MPO-DNA) and commercial ELISA (H3Cit). Correlations were tested using Spearman’s rho. Results There were no correlations between any of the circulating NET markers and gene expression of the fibrinolytic markers tPA, uPA and PAI-1 in the thrombus. Of the circulating NET markers, dsDNA correlated with thrombus gene expression of PAD4 (rs=0.491 p=0.006), whereas none of the NET markers correlated with gene expression of PAD4 in circulating leukocytes. Conclusion Circulating NET markers were not associated with gene expression of fibrinolytic activity in thrombi from patients with STEMI. Local NET formation in the thrombi, measured as PAD4 expression, was associated with the circulating NET marker dsDNA. These findings suggest that circulating NET markers have limited utility for assessing local fibrinolytic activity, but might reflect PAD4-dependent NET formation in coronary thrombi.

Assessment of Artemisia campestris L. Leaf Extract Effects on Polycystic Ovarian Syndrome in Rats, Antioxidant and α-Amylase Inhibition Activities.

Polycystic Ovarian Syndrome (PCOS) is characterized by metabolic and reproductive dysfunction, often associated with elevated oxidative stress markers in the bloodstream. This study examines the potential antioxidant properties and α-amylase inhibitory activity of Artemisia campestris leaves extract (Artemisia campestris L) and its effects on rats with induced PCOS. Estradiol valerate was administered to ten mature Wistar rats to induce PCOS, while a control group consisted of five mature Wistar rats. Following a 16-day induction period, the rats were categorized into three groups: a control group, a PCOS group, and an experimental group receiving 200 mg/kg body weight of A. campestris L. extract orally for 15 days. Serum levels of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) were measured using the ELISA technique. The group treated with A. campestris L. extract exhibited significantly reduced LH levels compared to the PCOS group. Histomorphometric analysis indicated notable changes in follicle counts and the thickness of the theca layer. These findings suggest a significant alleviation of PCOS symptoms, potentially linked to the effects of A. campestris L. on oxidative stress pathways. Furthermore, aqueous extracts of A. campestris L. displayed potent in vitro inhibition of α-amylase, with an IC50 value of 2.418 µg/mL.

Osteopontin as a biomarker for diagnosing the severity of osteoarthritis

Osteoarthritis (OA) is the most common type of arthritis. It is characterized by the loss of cartilage and pain ultimately leading to difficulty in movement. Glycoprotein osteopontin (OPN) is an essential regulator that plays a critical role in developing osteoarthritis. The study aims to determine the level of OPN in the serum of patients with OA and its correlation with the content of calcium (Ca), phosphorus (P), and magnesium (Mg). The study was conducted on 92 male and female patients aged between 30 and 65, categorized into mild, moderate, and severe groups through clinical examination and X-rays. The control group consist of 58 healthy males and females 30-65-year old. OPN was measured using the ELISA technique, and minerals were estimated using spectrophotometry. It was shown that in OA patients, OPN level increased highly significant when compared with the control group (10.7 ± 3.4 ng/ml) and depended on OA severity. In patients with severe OA, it was higher (47.5 ± 8.1 ng/ml) than in groups with moderate (14.8 ± 4.5 ng/ml) and mild (12.1 ± 3.1 ng/ml) extent of the disease. We showed a significant positive relationship between OPN and phosphorus levels and a negative significant correlation between OPN and calcium, calcium/phosphorus ratio, and magnesium. These findings underscore the potential of OPN as a valuable biomarker for diagnosing the severity of osteoarthritis and monitoring the effectiveness of treatment. Keywords: calcium, magnesium, osteoarthritis, osteopontin, phosphorus.

Contribution to a study on the seroprevalence of Equine Infectious Anemia in Tiaret province northwestern Algeria

A serological survey was conducted in equines in the province of Tiaret, in order to estimate the prevalence level of infection of equine infectious anemia (EIA). This survey has allowed to study the rate of infection by EIAV equine, to clarify the current situation of the EIA in the province of Tiaret, constitute a first step towards a national approach to control the emergence of this disease. Serum samples were collected from 293 horses and donkeys 30 and tested by ELISA technique. All results obtained by the ELISA test proved totally negative for all the samples studied. The equine infectious anemia is a topical disease. It is not well known and rarely sought on the ground. Yet the consequences of an outbreak can be very heavy on the economy.

Toxoplasma Gondii In Association with TLR4 Among Regular Hemodialysis Patients

Background: Hemodialysis patients can be considered immunocompromised as they have an increased risk of infections. The activity of Toll -like receptors in defense against infections was observed particularly for Toll like receptor4 molecule. Several single-nucleotide polymorphisms (SNPs) residing in genes encoding Toll -like receptors were reported as significant genetic modifications associated with infections. Aim: To determine the association between Toll- like receptors 4 polymorphisms and the susceptibility to infection with Toxoplasma gondii in hemodialysis patients. Materials and Methods: This case- control study was conducted on 150 patients with hemodialysis and 150 apparently healthy blood donors as control group, the samples were collected in the period from march 2021 to November 2021 from Al-Karama hospital and Al-Yarmouk Teaching Hospital. All patients were investigated for the presence of Toxoplasma gondii and Abs (IgG, IgM) by using ELISA technique and conformation by molecular technique (Real time-PCR). The selected SNPs in Toll- like receptors 4 were amplified by using conventional polymerase chain reaction PCR and then sequencing there polymorphism. Results: In the current study the comparison of median age between patients and controls was with no significant difference, while the laboratory tests revealed significant difference in median level of urea, creatinin , calcium , potassium ,albumin and hemoglobin, and insignificant difference for body mass index and sodium between studied groups. The ratio of females infected with Toxoplasma gondii was higher than infected males. The seropositive result in patients for Toxoplasma gondii - IgG and IgM was 15 and 6 respectively while 19 patients were positive by PCR. However in control group there were 8 patients seropositive for IgG and positive in PCR results while the results of IgM and PCR were negative for all control group. The Polymerase chain reaction (PCR) products of Toll- like receptors 4 genes were subjected for direct sequencing by using Bio edit software. And the resultant sequences were compared with reference sequences in NCBI. The analysis of TLR-4 rs4986790 genotypes were 24(80%) AA and 6(20%) AG in patients while 6 (60%) AA and 4(40%) AG in controls with insignificant difference, while the analysis of TLR-4 rs4986791 genotypes were 24(80%) CC and 6 (20%) CT in patients while 6 (60%) CC and 4(40%) CT in controls with insignificant difference. Conclusion: There was no significant association of Toxoplasma gondii with TLR4 genotypes of rs4986790 and rs4986791 SNPs in hemodialysis patients.

T-Cell Immunoglobulin and Mucin Domain 3 (TIM-3) Gene Expression as a Negative Biomarker of B-Cell Acute Lymphoblastic Leukemia.

B-cell acute lymphoblastic leukemia (B-ALL) accounts for 85% of all childhood ALL. Malignancies exhaust T and B cells, resulting in an increased expression of immune checkpoint receptors (ICRs), such as T-cell immunoglobulin and mucin domain 3 (TIM-3). TIM-3 has been found to be dysregulated in different types of cancer. However, there is a lack of rigorous studies on the TIM-3 expression in B-ALL. The current study aimed to measure the expression of TIM-3 at the gene and protein levels and evaluate the potential of TIM-3 as a biomarker in B-ALL. A total of 28 subjects were recruited between 2021 and 2023, comprising 18 subjects diagnosed with B-ALL and 10 non-malignant healthy controls. The B-ALL patients were divided into three groups: newly diagnosed (four patients), in remission (nine patients), and relapse/refractory (five patients). The expression levels of TIM-3 were evaluated using the real-time qPCR and ELISA techniques. The results revealed that the TIM-3 expression was significantly downregulated in the malignant B-ALL patients compared to the non-malignant healthy controls in the mRNA (FC = -1.058 ± 0.3548, p = 0.0061) and protein blood serum (p = 0.0498) levels. A significant TIM-3 gene reduction was observed in the relapse/refractory cases (FC = -1.355 ± 0.4686, p = 0.0327). TIM-3 gene expression allowed for significant differentiation between patients with malignant B-ALL and non-malignant healthy controls, with an area under the curve (AUC) of 0.706. The current study addressed the potential of reduced levels of TIM-3 as a negative biomarker for B-ALL patients.

Substance P a potent prognostic indicator for canine mammary tumours- a comparative study

Substance P, a neurotransmitter in the central and peripheral nervous system, is considered as a biomarker of pain due to its key role in nociception. But recent researches have proved the increased expression of Substance P and its receptors in human breast cancers and their role in its progression. Canine mammary glands and spontaneously occurring neoplasms in them are considered best models for human breast cancers. Hence as a preliminary study, the present research work was undertaken to assess the level of Substance P in dogs with mammary tumours in comparison to healthy dogs. From the dogs brought for ovariohysterectomy and mammary tumour resection, 20 female dogs were selected for the study. Signalment and detailed history regarding each dog were recorded. All the dogs were subjected to thorough preanaesthetic evaluation and were grouped into Group I (mammary tumour) and II (control group) with 10 dogs each. Blood was collected from all the dogs and serum was separated and stored at -20ºC for estimation of substance P. The same was performed using sandwich ELISA technique with canine specific Substance P ELISA kit. The values obtained for Group I and II were statistically analysed using independent t- test. The level of substance P was significantly higher in the dogs of Group I when compared to Group II Keywords: Substance P, mammary tumour, dogs

Evaluating Antioxidant Enzyme Parameters in Rats Treated with Berberine-Loaded Bovine Serum Albumin Nanoparticles and Scopolamine

Abstract Introduction/Objective Introduction: Berberine (BER) has garnered attention for its antioxidant properties demonstrated in various studies, showing its ability to scavenge or inhibit reactive oxygen species (ROS) and reactive nitrogen species (RNS), including nitric oxide, superoxide anion, and peroxynitrites. Methods/Case Report Methods: Forty-two male Wistar rats were randomly assigned to seven groups, each comprising 6 rats: Sham control (no treatment), BSA-NPs orally administered, BRB-NPs orally administered, SCO intraperitoneally injected, BER followed by SCO injection, BSA-NPs with SCO injection, and BER-NPs with SCO injection. Treatments were administered daily for 28 days. The cholinergic status was evaluated by assessing acetylcholinesterase (AChE) activity in brain homogenates, and brain homogenate Aβ-42 levels were measured using the ELISA technique. Additionally, brain prooxidant markers (NO, TBARS, XO), nonenzymatic antioxidants (GSH), and enzymatic antioxidants (GPx, GST, and SOD) were measured. Results (if a Case Study enter NA) Results: The study revealed that treatment with BRB NPs resulted in the lowest AChE activity and Aβ-42 levels. Scopolamine administration significantly increased brain NO, TBARS, and XO activity, accompanied by a progressive depletion in GSH content (p&amp;lt;0.05). Treatment with BRB-free or BSA decreased NO levels, while the BSA or BRB-NPs groups exhibited higher brain TBARS levels than the sham group. All treated groups exhibited similar activity to the sham control, except for the BSA-treated group. Furthermore, all treated groups showed comparable GSH levels to the sham control, except for the scopolamine group. Treatment with BRB NPs normalized GPx activity and showed the highest GST activity. The BRB-NPs treated group also exhibited the highest SOD level. The p-value was &amp;lt;0.001 among all studied groups. Conclusion Conclusion: Injection of SCO induced oxidative impairments in brain tissue, evidenced by increased prooxidants such as NO and depletion of antioxidant capacity (GPx, GST, and GSH). SCO also disrupted glutathione metabolism, enhancing the level of lipid peroxidation in the brain. Treatment with BRB-NPs or free BRB progressively increased antioxidant capacity and mitigated the harmful effects of oxidative damage in the brain by reducing prooxidant levels.