Enzyme-linked Immune Sorbent Assay Research Articles

Insulin Resistance in Chronic Hepatitis C Non Diabetic Patients

Background: Patients with chronic hepatitis have impaired glucose metabolism with hyperinsulinemia and insulin resistance, this hyperinsulinemia has been shown to be due to decreased insulin catabolism rather than increased pancreatic insulin secretion. We aimed to evaluate insulin resistance in non-diabetic patients with chronic hepatitis C virus infection. Subjects and methods: The study was a case-control study conducted in Tropical Medicine and Gastroenterology Department King Abdullah University Hospital. 60 patients and 30 healthy controls were included in the study. The patients were classified into two groups: Group A: 30 patients with chronic hepatitis C infection were selected with positive HCV RNA in serum for at least 6 months; Patients were not receiving anti-viral therapy at the time of sampling. They showed no evidence of cirrhosis. Group B: 30 patients with HCV related liver cirrhosis. They were divided according to Child Pugh score; twenty patients with HCV related compensated liver cirrhosis (Child A). Ten patients with HCV related decompensated liver cirrhosis (Child B and C). Group C: The control group: included 30 healthy individuals. All patients and controls were subjected to the following: Liver function tests: Alanine transaminase (ALT), Aspartate transaminase (AST), total and direct bilirubin, total protein, serum albumin. Prothrombin time (PT) & international normalization ratio (INR). Renal function tests: Blood urea nitrogen (BUN), Na, K. Complete blood count. Alpha fetoprotein (αFP). Diagnosis of chronic hepatitis C infection was based on positive HCV by PCR, persistent elevation of liver enzymes more than 6 months and liver biopsy for some of the patients. Anti-hepatitis C virus antibody (HCV Ab) using third generation enzyme linked immune sorbant assay (ELISA) test., Hepatitis B virus (HBV): HBVsAg., Overnight fasting and two hours postprandial blood glucose level. Fasting serum insulin of everyone. Insulin resistance was determined via the Homeostasis Model assessment (HOMA-IR) by the following equation: - Insulin resistance: Fasting insulin (μu/ml) x Fasting glucose (mmol/L) 22.5 An index value of > 2.5 was defined as IR. This cutoff value was chosen because studies suggested that a HOMA-IR of 2.4-3.0 is probably suitable to define IR in CHC patients. Blood samples were collected after 12 hours of overnight fasting. Results: We found that out of 30 CHC and 30 LC (20 compensated LC, 10 de compensated LC) 8 (26.7%); 8 (40%) patients and 5(50%) respectively had HOMA-IR levels greater than 2.5, which is consistent with IR diagnosis. Decompensated cirrhotic patients showed higher frequency of IR compared to CHC and compensated cirrhotic patients. Conclusion: In chronic hepatitis C patients, HOMA-IR, fasting serum insulin and fasting blood glucose were significantly higher than healthy controls (p <0.0001).

Experimental-based mechanobiological modeling of the anabolic and catabolic effects of breast cancer on bone remodeling.

Bone is a biological tissue characterized by its hierarchical organization. This material has the ability to be continually renewed, which makes it highly adaptative to external loadings. Bone renewing is managed by a dynamic biological process called bone remodeling (BR), where continuous resorption of old bone and formation of new bone permits to change the bone composition and microstructure. Unfortunately, because of several factors, such as age, hormonal imbalance, and a variety of pathologies including cancer metastases, this process can be disturbed leading to various bone diseases. In this study, we have investigated the effect of breast cancer (BC) metastases causing osteolytic bone loss. BC has the ability to affect bone quantity in different ways in each of its primary and secondary stages. Based on a BR mathematical model, we modeled the BC cells' interaction with bone cells to assess their effect on bone volume fraction (BV/TV) evolution during the remodeling process. Some of the parameters used in our model have been determined experimentally using the enzyme-linked immune-sorbent assay (ELISA) and the MTT assay. Our numerical simulations show that primary BC plays a significant role in enhancing bone-forming cells' activity leading to a 6.22% increase in BV/TV over 1 year. On the other hand, secondary BC causes a noticeable decrease in BV/TV reaching 15.74% over 2years.

The predictive diagnosis for ANGPTL8 level in women with T2DM

Background: The metabolic syndrome including type 2 diabetes mellitus(T2DM), insulin resistance(IR), obese, hypertension, and dyslipidemia, is an increasing health burden. In the last decade, various metabolic hormones have been noticed to significantly role obese and associated complications. for example of this, the adipokines or hepatokines secreted proteins from hepatocyte and adipocyteare correlate with metabolic syndrome. Angiopoietin-like protein 8 (ANGPTL8) has a novel hepatokine or adipokine, which is mostly expressed in the adipose tissue, and liver stimulates the β-cell proliferation as a pancreatic in an insulin resistance state. It's a recently discovered the endocrine regulator associated with glucose homeostasis, IR, and metabolism of lipid. Materials and Methods: Ninety women participated in this study, comprising sixty patients with diabetes mellitus type-2 and thirty healthy (sex and age - matched) as a control group. Enzyme linked immune sorbent assay (ELISA) was used to assess ANGPTL8 and insulin. Standard procedures were used to measure fasting blood glucose, lipid profile, and glycated hemoglobin (HBA1c) in a certified laboratory. Results: Diabetes patients had higher levels of ANGPTL8 than the control group. The blood ANGPTL8 level in the Type-2 diabetes group varied from 4432.89±1171.09 pg/ml, while the control group's level was 2892.11±537.91pg/dl (p<0.0001).

Association of circulating miRNA-21-5p and miRNA-146a-5p expression level with disease activity of ankylosing spondylitis

BackgroundThe etiopathogenesis of ankylosing spondylitis (AS) involve interaction between genetic and environmental factors which exert an effect on epigenetic mechanisms including non-coding miRNAs causing immune dysfunction and inflammation. This study is designed to find if there is an association in the expression levels of miRNA- 21-5 (miR-21-5p) and miR- 146a-5p with the disease activity, also to assess their role in the follow up of patients. MethodSixty individuals enrolled in this case-control study; 30 individual (28 males and 2 females) as patients group, and the healthy controls group include 30 individuals (27 males and 3 female). Twenty four patients were human leukocyte antigen (HLA B-27) positive, while the other 6 patients were negative. Reverse transcription polymerase chain reactions (RT- PCR) was used for determination of the expression levels of miRNA-21-5p and miRNA-146a-5p in serum of patients and controls, while concentration of high sensitivity C- reactive protein (HS- CRP) was estimated using enzyme linked immune-sorbent assay (ELISA). These tests were done two times for each patient; the first is before starting biologic treatment and the second after two months from taking the first dose. ResultsThere are significant elevations (p < 0.001) in the expression of miR- 21-5p and miR- 146a-5p also in the serum level of Hs-CRP in patients compared with controls. In patient group, miR-21-5p expression is significantly reduced at end of the study (p value <0.05), while there is no significant difference (p > 0.05) of miR-146-5p expression between baseline and end of the study. At base line of the study; it is found that the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and Bath Ankylosing Spondylitis Functional Index (BASFI) scores and Hs-CRP were positively correlated with both of miR- 146a-5p and miR- 21-5p and the correlation remain positive at in the end of study. In conclusion, expression level of circulating miR-21-5p and miR-146a-5p could be serve as non invasive biomarkers for the diagnosis of AS. Micro-RNA-21-5p associated with the activity and severity of the disease and it could be used for follow up patients with AS and for assessment the response to biological treatments.

Prevalence of Mycoplasma bovis Infection in Calves and Dairy Cows in Western Australia.

Simple SummaryMycoplasma bovis is an emerging pathogen of economic and welfare concern for both adult and young cattle. A study was conducted to determine the prevalence of M. bovis in adult cows and calves in the southwest region of Western Australia. Nasal swabs and blood samples were collected from the animals and bulk tank milk samples were assessed for both seroprevalence and active infections of M. bovis infections in adult cows and calves. The study recorded a high seroprevalence of M. bovis in 699 apparently healthy adult lactating cows and 495 young calves on 29 dairy farms. The herd-level seroprevalence was also detected as being higher in both adult lactating cows and calves. No current active infections were recorded on the farms. The female calves and pure Holstein–Friesian animals were found to be twice as likely to be seropositive for M. bovis compared to male calves and the Holstein–Friesian crossbred calves. The high seroprevalence of M. bovis in both adult and young cattle in the southwest dairy farms of Western Australia warrants more effective farm biosecurity measures and further evaluation of the current prevention and management measures practiced on the farms.Mycoplasma bovis (M. bovis) can cause a multitude of diseases in cattle, with detrimental effects on the farm economy and the welfare of both adult and young cattle. The objective of this study was to determine the prevalence of M. bovis in adult cows and calves in the south-west region of Western Australia. A cross-sectional study was conducted on 29 dairy farms with 699 apparently healthy adult lactating cows and 495 young calves during 2019–2020. Nasal swabs and blood samples collected from the animals and bulk tank milk (BTM) samples were assessed for M. bovis-specific proteins and antibodies by using polymerase chain reaction (PCR) and Mycoplasma immunogenic lipase A- Enzyme-Linked Immune Sorbent Assay (MilA ELISA). A seroprevalence of 42.5% (95% CI: 38.9–46.2) and 61% (95% CI: 56.6–65.2) was found in adult lactating cows and calves, respectively. The herd-level seroprevalence of M. bovis ranged from 4% (95% CI: 07–19.5) to 92% (95% CI: 75.0–97.8) in adult lactating cows and 25% (95% CI: 10.2–49.5) to 87% (95% CI: 67.9–95.5) for calves in these farms. None of the BTM and nasal swab samples were positive for M. bovis, indicating an absence of any current active infections on the farms. The female calves and pure Holstein–Friesian animals are twice as likely to be seropositive for M. bovis compared to male calves (OR 2.4; 95% CI: 1.7–3.5) and Holstein–Friesian crossbred calves (OR 2.4; 95% CI: 1.7–3.5). The high seroprevalence in both adult and young cattle in the southwest dairy farms of Western Australia warrants more effective farm biosecurity measures and further evaluation of the current prevention and management measures practiced on the farms.

Electroacupuncture at Lower He-Sea and Front-Mu Acupoints Ameliorates Insulin Resistance in Type 2 Diabetes Mellitus by Regulating the Intestinal Flora and Gut Barrier.

IntroductionThe study objective was to investigate the effects of electroacupuncture performed at the he-sea and front-mu acupoints on the intestinal microflora and intestinal barrier in db/db mice and to explore the related mechanism in type 2 diabetes mellitus.MethodsDb/m mice in the normal control group (NOC), electroacupuncture group (EA), metformin group (MET) and T2DM group (T2DM) were used as model controls, and db/db mice were used in all three groups, with 8 mice in each group. The treatment period was 2 weeks. Fasting blood glucose (FBG) and triglyceride (TG) levels were measured. Lipopolysaccharide (LPS) and tumor necrosis factor-α (TNF-α) levels were detected by enzyme-linked immune sorbent assay (ELISA). The ileal tissue was stained with hematoxylin-eosin staining (H&E), and histopathological changes were observed under a light microscope. Illumina sequencing was used to analyze the V4 region of the 16S rRNA gene to evaluate the effect of EA on the intestinal flora.ResultsOur results suggest that EA treatment can reduce the expression of diabetes-related markers, with an effect similar to that of metformin. After EA intervention, the abundance of Firmicutes and the ratio of Firmicutes to Bacteroidetes increased, while the abundances of Bacteroidetes and Eubacterium decreased. In addition, the serum levels of LPS and TNF-α in the electroacupuncture group were downregulated, and ileal tissue damage was alleviated under an electron microscope.ConclusionEA combined with acupoints can restore the intestinal flora structure, decrease the blood LPS level, reduce levels of inflammation, maintain the integrity of the intestinal barrier, and play a therapeutic role in the treatment of T2DM, mainly by increasing the abundance of Firmicutes and the ratio of Firmicutes to Bacteroidetes and decreasing the abundances of Bacteroidetes and Eubacterium.

Irisin Level in Salah Adin’s Insulin Resistance Syndrome Patients in View of Global Controversy

Background: Metabolic syndrome (high blood pressure, hyperglycemia, dyslipidemia, and obesity) is now an international health problem that impacts about 20 per cent to 25 per cent of the total population and can be worsened in the developing world, particularly in the Middle East, which is roughly two out of every five Iraqis. The metabolic syndrome has a direct link to many life-threatening diseases, such as cardiovascular disease, early onset of some cancers, such as colorectal cancer, Chronic liver illnesses, polycystic ovarian syndrome, and others. Methodology: A case-control study on a total of ninety volunteers (46 males and 44 females), comprising sixty metabolic syndrome patients (25 males and 35 females), defined in terms of international diabetic federation criteria, and thirty controls, is to be conducted at Salah Alden general hospital. Adults men and women aged (24-to 65) years who attended Salah Alden's main hospital outpatient clinics in Tikrit\Iraq, from October 2021 to March 2022 were included in the research. The levels (Irisin, and Insulin) in all research groups and controls are evaluated using an Enzyme-Linked Immune-sorbent Assay. Results: Irisin level was significantly higher among controls with a mean of (6.84ng/ml), compared to metabolic syndrome patient mean (5.71 ng/ml), and significantly negatively correlated to triglycerides level, and non-significantly negatively correlated with (systolic blood pressure, fasting serum glucose, and waist circumference) and non-significantly positively correlated to high-density lipoprotein level. Irisin levels were significantly different among treatment-based groups with the highest mean level among the treatment-free group (6.8450ng/ml±1.96052), followed by the antihypertensives taking group (5.3340ng/ml±2.99273), and the lowest mean level was recorded among diabetics (4.8916ng/ml±2.81952). Conclusion: With the exception of HDL, a high level of Asprosin and a low level of Irisin are strongly linked to metabolic syndrome and its components. Irisin levels differ amongst metabolic syndrome patients with various therapy groups. Keywords: Irisin, metabolic syndrome, Insulin Resistance.

The Relationship Between Cardiovascular Disease Risk Score and Postoperative Delirium: The PNDABLE Study

ObjectiveWe aimed to investigate the relationship between Framingham Heart Study general cardiovascular disease risk score (FHS–CVD risk score) and postoperative delirium (POD) among patients who had unilateral total knee arthroplasty performed under epidural anesthesia. Furthermore, we examined whether such a hypothesized relationship was mediated by the cerebrospinal fluid (CSF) biomarkers.MethodsA total of 750 participants were included in the current study. And the data were drawn from the database obtained from the Perioperative Neurocognitive Disorder And Biomarker Lifestyle (PNDABLE) study. The preoperative cognitive function of participants was measured by using Mini-Mental State Examination (MMSE). The incidence of POD was assessed using the Confusion Assessment Method (CAM). The POD severity was measured using the Memorial Delirium Assessment Scale (MDAS). The POD CSF biomarkers included in the current study were: Aβ42, T-tau, P-tau, Aβ42/T-tau, and Aβ42/P-tau. The level of the CSF biomarkers was measured using the enzyme-linked immune-sorbent assay (ELISA) in the PNDABLE study. Linear regression analysis was performed to examine the relationship between the FHS–CVD risk score and the POD CSF biomarkers. Logistic regression was used to analyze the relationship between FHS–CVD risk score, POD CSF biomarkers, and POD incidence. The proposed mediating effect of CSF biomarkers was evaluated using Mediation Analysis with 10,000 bootstrapped iterations. The receiver operating characteristic (ROC) curve is chosen as the evaluation metric for assessing the efficacy of the FHS–CVD risk score in predicting POD.ResultsIn the PNDABLE study, the overall incidence of POD was 22.9% with 37.2% in the higher vascular risk group and 7.9% in the lower vascular risk group. Multiple linear regression models showed that a higher preoperative FHS–CVD risk score was positively correlated with CSF T-tau (β = 0.218, P = 0.015) and P-tau level (β = 0.309, P < 0.001) in the higher vascular risk group. After adjusting for age (40–90 years), gender, education, MMSE, smoking history, drinking history, hypertension, diabetes, and the presence of CHD (cardiovascular heart disease), the results of the logistic regression analysis demonstrated the effect of Aβ42 (OR = 0.994, 95% CI 0.992–0.996, P < 0.001), Aβ42/T-tau (OR = 0.353, 95% CI 0.254–0.491, P < 0.001), and Aβ42/P-tau (OR = 0.744, 95% CI 0.684–0.809, P < 0.001) in protecting patients against POD. However, the FHS–CVD risk score (OR = 1.142, 95% CI 1.017–1.282, P = 0.025) and the remaining two biomarkers: T-tau (OR = 1.005, 95% CI 1.004–1.007, P < 0.001) and P-tau (OR = 1.045, 95%CI 1.029–1.062, P < 0.001) were identified as the risk factors. Mediation analyses revealed that the association between FHS–CVD risk score and POD was partially mediated by T-tau (proportion: 31.6%) and P-tau (proportion: 23.6%). The predictive power of the FHS–CVD risk score was validated by the ROC curve with an AUC of 0.7364.ConclusionHigher vascular risk score is one of the preoperative risk factors for POD, which is partly mediated by CSF biomarker tau protein.Clinical Trial Registration[www.clinicaltrials.gov], identifier [ChiCTR2000033439].

Assessment of plasmin and inducible nitric oxide synthase in the patients with COVID-19

Aims of study: The purpose of this study is to compare the levels of plasmin, inducible nitric oxide, and nitric oxide in COVID-19 infection patients to a healthy control group, with a scientific interpretation of the predicted results. Methods: Data was collected from ninety people, divided into two groups: patients and controls, and a demographic study was conducted, which included age, gender, oxygen saturation, and BMI, with diabetes and asthma eliminated. The plasmin and inducible nitric oxide levels are estimated using the enzyme-linked immune sorbent assay (ELISA) technique, and nitric oxide is measured using a spectrophotometer. Results: Patients with COVID-19 showed a substantial rise in the values of plasmin and inducible nitric oxide synthase enzyme, as well as a decrease in the value of nitric oxide, when compared to healthy controls.

A preliminary study on aflatoxin exposure by urine biomonitoring in Chile.

We assessed the occurrence of aflatoxin M1 (AFM1) in urine in a sample of the MAUCO population-based cohort (n = 120) using an enzyme-linked immune sorbent assay (ELISA) kit specially designed for the analysis of AFM1 in urine. We found AFM1 in the urine of 59% of the participants (> limit of detection), with 12% of the samples being over the limit of quantification. The mean of the quantifiable samples was 0.66 (± 0.35) ng/mg adjusted creatinine, ranging from 0.31 to 1.39ng/mg creatinine. The mean probable daily intake (PDI) of AFB1 was 0.23 (± 0.37) ng/kg bw according to the upper bound (UB), being significantly higher in women and 0.14 (± 0.23) ng/kg bw in the modified lower bound (mLB) approach, ranging from 0.01 to 1.98ng/kg bw. The risk of AFB1 was assessed with the margin of exposure (MOE) approach estimated at 2800 in the mean mLB and 1733 in the mean UB. According to the MOE values obtained in this study, aflatoxin B1 exposure must be considered a public health concern and must be taken as a priority for food risk management.

Krebs von den Lungen-6 (KL-6), soluble programmed cell death-1 (sPD-1) and its ligand-1(sPDL-1) in systemic sclerosis patients: Relation to disease parameters

Aim of the workThis study aimed to assess the levels of Krebs von den Lungen-6 (KL-6), soluble programmed cell death-1 (sPD-1) and soluble programmed cell death ligand-1(sPDL-1) in systemic sclerosis (SSc) patients, and to evaluate their relation with disease parameters. Patients and methodsForty-six SSc patients (31 limited, 15 diffuse) and 43 matched controls were included. Serum levels of KL-6, sPD-1, and sPDL-1 were assessed by enzyme-linked immune-sorbent assay (ELISA). ResultsPatients mean age was 40.2 ± 12.2 years, disease duration 8.04 ± 6.02 years and females: male was 10.5:1. Levels of KL-6, sPD-1, and sPDL-1 were significantly higher in SSc patients compared to the controls 22.4(14.1–637.6) vs. 14.8(8.9–182.3) (p < 0.001), 3.1(1.8–64.2) vs. 1.2(0.68–14.4) (p < 0.001), and 2.8(1.5–84.4) vs. 1.4(0.92–36.4) (p < 0.001) respectively. KL-6 and sPD-1 levels were significantly lower in diffuse SSc subtype than the limited subtype 20.7(14.1–637.6) vs. 24.5(16.2–328.8) (p = 0.01), and 2.5(1.8–64.2) vs. 3.2(2.1–63.4) (p = 0.03) respectively. KL-6 was significantly decreased in SSc patients who developed pulmonary hypertension compared to those without 20.9(14.1–36.3) vs. 27.5(16.5–637.6) (p = 0.02), and in those who received cyclophosphamide 21(14.1–310.9 vs. 31.6(16.2–637.6) (p = 0.019). The three markers were significantly correlated. KL-6 levels with sPD-1 (r = 0.79, p < 0.001) and sPDL-1 (r = 0.79, p < 0.001) and between sPD-1 and sPDL-1 (r = 0.82, p < 0.001) respectively. Both sPD-1 and sPDL-1 significantly correlated with age (r = 0.29, p = 0.048 and r = 0.34, p = 0.021 respectively). ConclusionsKL-6, sPD-1, and sPDL-1 may be considered as potential biomarkers in the diagnosis and assessment of SSc patients. Significantly lower levels of KL-6 were detected in diffuse SSc patients, those with pulmonary hypertension, and patients who received cyclophosphamide.

Vitamin D as predictor Marker of kidney disease in males with type 2 diabetes mellitus

The current work was done to determine the correlation between vitamin D and cystatin C as a predictor of kidney disease in males with diabetes mellitus. A total of 60 males were taken from October to December 2018. They were divided into 30 patients with type 2 diabetes and 30 healthy volunteers' as a control group. Glycated hemoglobin HbA1c, plasma glucose level, creatinine and urea were measured for all subjects. Also, levels of 25(OH) D and cystatin-c were determined using the enzyme-linked immune sorbent assay method. In this current study, as expected, patients with diabetes mellitus had significantly higher (p&lt; 0.001) fasting blood glucose (FBG), body mass index (BMI), and glycated hemoglobin (HbA1c) levels as compared to the age-matched controls group. In addition, a significantly higher increase in the average level of urea, creatinine, and cystatin-c while significant higher decreases in vitamin D concentration compared to the patients group with the control group were found. A significant negative correlation was found between 25(OH) D and urea levels. On the other hand, a significant positive correlation was observed between the 25(OH) D level and HbA1c, FBG, cystatin-c and creatinine. From the results of this work, vitamin D could be a valuable predictor of nephropathy in males with diabetes mellitus. Likewise, further work is required to suggest that vitamin D may be prone to nephropathy in all patients with diabetes mellitus by estimating cystatin C as a clinical risk for kidney function. Keywords: Nephropathy, cystatin C, vitamin D.

Maternal serum telomerase levels increase in pregnancies with mild and severe preeclampsia

IntroductionConsidering that preeclampsia is characterized by oxidative stress, inflammation, and endothelial dysfunction, we hypothesized that preeclampsia and preeclampsia severity may affect the telomerase levels of the mother. MethodsThis cross-sectional case control study comprised 86 participants who were separated into three groups as severe preeclampsia, non-severe preeclampsia, and healthy control group. Venous blood samples were obtained from pregnant women with severe preeclampsia just before delivery for biochemical analysis and to evaluate maternal serum telomerase levels. Since gestational age, maternal age and BMI would have an effect on maternal serum telomerase levels, serum samples were taken in the control group and non-severe preeclampsia group at similar gestational age during clinical visits in order to homogenize these parameters, and these patients were followed up. Telomerase levels in maternal serum were evaluated using the enzyme-linked immune-sorbent assay. ResultsMaternal age, nulliparity, body mass index (kg/m2) at blood sampling day, ethnicity, smoking and history of caesarian section were statistically similar among the groups. The mean birth weight percentiles were the lowest in the severe preeclampsia group. Fetal growth restriction rates were significantly higher in the severe preeclampsia group than in the non-severe preeclampsia group. Gestational age at blood drawn was similar among groups. Neutrophil lymphocyte ratio, platelet lymphocyte ratio, mean platelet volume, red cell distribution width and white blood cell were statistically different among groups. The serum telomerase level was 1.137 ± 0.390 ng/mL in the severe preeclampsia group, 0.763 ± 0.390 ng/mL in the non-severe preeclampsia group, and 0.425 ± 0.160 ng/mL in the control group (p < .001). DiscussionThis study indicated that maternal serum telomerase levels were significantly increased in both preeclampsia groups.

Punicalagin suppresses inflammation in ventilator-induced lung injury through protease-activated receptor-2 inhibition-induced inhibition of NLR family pyrin domain containing-3 inflammasome activation.

Punicalagin is recorded to be a potent anti-inflammatory drug, while its effect on inflammation existing in ventilator-induced lung injury (VILI) requires further verification. Rats were pretreated with punicalagin, followed by VILI modeling. Lung histopathological examination was performed with hematoxylin-eosin staining accompanied by the lung injury score. The lung wet/dry (W/D) weight ratio and total bronchoalveolar lavage fluid (BALF) protein level were measured. After transfection with protease-activated receptor-2 (PAR2) overexpression plasmids, mouse alveolar epithelial MLE-12 cells were treated with punicalagin and then subjected to cyclic stretching. Punicalagin's cytotoxicity to MLE-12 cells were measured by MTT assay. The levels of inflammatory cytokines (tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6), PAR2, NLR family pyrin domain containing-3 (NLRP3), and apoptosis-associated speck-like protein containing a CARD (ASC) in the BALF, lung tissues or cells were analyzed by enzyme-linked immune-sorbent assay (ELISA), qRT-PCR or/and western blot. Punicalagin treatment attenuated VILI-induced lung histopathological changes and counteracted VILI-induced increases in the lung injury score, W/D weight ratio and total protein level in BALF. Also, punicalagin treatment counteracted in vivo VILI/cyclic stretching-induced increases in the levels of PAR2, inflammatory cytokines, NLRP3, and ASC. PAR2 overexpression potentiated the cyclic stretching-induced effects, while punicalagin treatment revoked this PAR2 overexpression-induced potentiation effect. In turn, PAR2 overexpression partly resisted the punicalagin treatment-induced counteractive effects on the cyclic stretching-induced effects. Punicalagin suppresses inflammation in VILI through PAR2 inhibition-induced inhibition of NLRP3 inflammasome activation.

Anti mullerian hormone and antral follicle count for prediction of ovarian reserve in female infertile patients

Ovarian reserve (OR)determines the quantity and quality of follicles present in the ovary. It is independent of menstrual cycle in women of reproductive age. The chances of conceiving in infertile women can be predicted by understanding and determining the OR. The aim of the study was to assess and determine whether Anti-Mullerian Hormone (AMH) with Antral Follicle Count (AFC) or AMH alone is a better predictor of ovarian reserve in female infertile patients. Methods: In this prospective, cross-sectional study, a total of 60 patients were measured for AMH levels of which 30 patients was assessed along with AFC. The demographic details were collected and 5ml of venous blood samples were taken on day 2 of menstrual cycle. Serum was separated and stored at -20oC. Serum AMH levels were analysed using enzyme-linked immune sorbent assay (ELISA) whereas Antral Follicle Count with ultrasonography. Finally, 54 patients completed the analysis with the mean age of 26.32±3.95 years and body mass index of 24.09±4.62. The average AMH levels and AFC were found to be 2.49±4.05 ng/ml and 14.92±8.69 follicles respectively. AMH showed negative correlation with both FSH and LH and no correlation with estradiol.

Seroprevalence of syphilis among Sudanese pregnant ladies in Berber city, Sudan

Background: Because syphilis is often asymptomatic, it might go unrecognized. Furthermore, infection transfer from mother to unborn child is a concern. Preventing poor pregnancy effects in relation with maternal infection by screening and treating women for syphilis throughout pregnancy. Aiming at exploring anti treponemal antibodies among pregnant women in Berber Teaching Hospital our study was implemented. Methods: A cross-sectional, hospital based study was performed at Berber Teaching Hospital in River Nile State. Encompassed pregnant women who attended Hospital. Following an explanation of the study's goal, 100 serum samples were taken, and data was collected from each respondent using an interview questionnaire, 100 blood samples were obtained to identify syphilis anti bodeis. All specimens were analyzed using the Rapid Plasma Reagin test (RPR), which was confirmed by an enzyme linked immune sorbent assay (ELISA) (fortress diagnostics/UK). Statistical Package for Social Sciences (SPSS version 19) was used to manage informational gathered. Results: ELISA found 9% of syphilis positive patients, whereas RPR identified 10%.Age of respondents ranged between 16 to 43 years, 53 % of study participants categorized at (30 – 36 years) and 62 % at third trimester of gestational. The highest prevalence of syphilis was 4 % reported in age group (23 – 29 years) and 5% at third trimester. Conclusion: The syphilis sero-prevalence among pregnant women in the current research was 9% by ELISA and 10% by RPR. More precise and sensitive procedures, such as PCR, must be used to corroborate the findings

Advanced detection and sensing strategies of Pseudomonas aeruginosa and quorum sensing biomarkers: A review.

Pseudomonas aeruginosa (P. aeruginosa), a ubiquitous opportunistic pathogen, can frequently cause chronic obstructive pulmonary disease, cystic fibrosis and chronic wounds, and potentially lead to severe morbidity and mortality. Timely and adequate treatment of nosocomial infection in clinic depends on rapid detection and accurate identification of P. aeruginosa and its early-stage antibiotic susceptibility test. Traditional methods like plating culture, polymerase chain reaction, and enzyme-linked immune sorbent assays are time-consuming and require expensive equipment, limiting the rapid diagnostic application. Advanced sensing strategy capable of fast, sensitive and simple detection with low cost has therefore become highly desired in point of care testing (POCT) of nosocomial pathogens. Within this review, advanced detection and sensing strategies for P. aeruginosa cells along with associated quorum sensing (QS) molecules over the last ten years are discussed and summarized. Firstly, the principles of four commonly used sensing strategies including localized surface plasmon resonance (LSPR), surface-enhanced Raman spectroscopy (SERS), electrochemistry, and fluorescence are briefly overviewed. Then, the advancement of the above sensing techniques for P. aeruginosa cells and its QS biomarkers detection are introduced, respectively. In addition, the integration with novel compatible platforms towards clinical application is highlighted in each section. Finally, the current achievements are summarized along with proposed challenges and prospects.

Hydroxysafflor yellow A protects against ulcerative colitis via suppressing TLR4/NF-κB signaling pathway.

Hydroxysafflower yellow A (HSYA) protects against acute kidney injury through TLR4/NF-κB pathway. However, the effect and potential mechanism of HSYA in ulcerative colitis (UC) have been rarely reported, which is thus investigated in this research. An in vivo UC model was established by oral administration of 5% dextran sulfate sodium (DSS) in Sprague-Dawley rats. After HSYA treatment, the daily body weight and colon length of rats were measured. Then rat colon tissues, myeloperoxidase (MPO) activity, and the levels of inflammatory cytokines were examined by histopathological examination (HE) staining, immunohistochemistry, ultraviolet spectrophotometry, and enzyme-linked immune sorbent assay (ELISA) respectively. The activated TLR4/NF-κB pathway was detected by Western blot. RAW 264.7 cell viability was detected by MTT assay after lipopolysaccharide (LPS) treatment, and ELISA and Western blot were performed again to investigate the effects of HSYA on LPS-treated cells. DSS administration increased body weight and colon length of rats and induced colon tissue injury. DSS or LPS treatment up-regulated the levels of TNF-α, IL-1β, and IL-6 and activated TLR4/NF-κB pathway of colon tissues and cells, respectively. HSYA partially reversed the above effect of DSS and LPS treatment, and the effects of the drug were improved with the dosage. Taken together, HSYA alleviates UC by suppressing TLR4/NF-κB signaling pathway, which may provide a new insight for the treatment of UC.

Follistatin dysregulation impaired trophoblast biological functions by GDF11-Smad2/3 axis in preeclampsia placentas

IntroductionPreeclampsia (PE) is one of the main causes of maternal, fetal, and neonatal mortality. So far, the underlying mechanism of this pregnancy-specific syndrome remains unelucidated. The expression of Follistatin (FST) decreased in maternal serum (especially early onset severe PE) and placental trophoblasts of PE patients. However, whether FST-deficiency in preeclamptic placentas alters trophoblast function remains to be determined. MethodsTrophoblast cell lines were cultured in vitro and LV3 short hairpin RNA (shRNA) was used to silence FST. Growth and differentiation factor 11 (GDF11) expression level in placentas and serum were detected by immunohistochemistry and enzyme-linked immune-sorbent assay, respectively. To verify the effect of reduced FST expression on trophoblasts, microRNA-24-3p, which was predicted to target the 3′-untranslated region (3′-UTR) of FST, was screened out, and miR-24-3p mimic, inhibitor was used to regulate FST expression in trophoblasts. ResultsDownregulation of FST significantly enhanced the apoptosis and impaired migration and invasion of trophoblast. Reduced FST caused the upregulation of GDF11 in trophoblasts. Interestingly, GDF11 reduced in preeclamptic placental microvascular endothelial cells. Dysregulation of FST-GDF11-Smad2/3 signaling pathway, leading to increased apoptosis of trophoblast. Expression levels of miR-24-3p, was significantly elevated in preeclamptic placentas. Trophoblast cells transfected with miR-24-3p mimics displayed impaired migration and invasion and increased apoptosis. Treated by miR-24-3p inhibitor, trophoblast cells exhibited rescued function. DiscussionFST-deficiency impaired trophoblast function by upregulating GDF11 levels in trophoblasts. The regulation of FST-GDF11-Smad2/3 axis by microRNAs mimic or inhibitor may be critical to trophoblast function regulation and helps to deepen our understanding of the molecular mechanism of PE.

Acute renal failure in scrub typhus patients

Scrub typhus, a bacterial infection caused by Orientia Tsutsugamushi (formally Rickettsia) due to bite of the mite, and is increasing every year, and becomes a important cause of acute febrile illnesses. Due to lack of specific diagnostic facilities in most rural setups and non-specific clinical presentations, it becomes a challenge for clinician to correctly diagnose it on clinical examination. Clinically it presents with non-specific febrile illnesses with constitutional symptoms such as fever, rash, myalgia and headache or with organ dysfunction involving kidneys, lungs, liver, central nervous system or with circulatory collapse with hemorrhagic features. Eschar is the characteristic lesion which is present at the bite site of mite. Later an ulcer with black necrotic centre and regional lymphadenopathy occurs. Renal involvement in the form of Acute Renal Failure is seen very occasionally in these patients. So this Case-control study is designed for a period of six months which are october-2018 to march-2019, comparing 80 ELISA(Enzyme Linked Immune Sorbent Assay) confirmed cases of scrub typhus and 50 healthy controls. Statistically significant difference in Urea, Creatinine and Uric acid levels were found between two groups, using 't'-test with the significance value of P&amp;#60;0.0001.