RNA methylation machinery and m6A target genes as circulating biomarkers of ulcerative colitis and Crohn's disease: Correlation with disease activity, location, and inflammatory cytokines.

Accurate diagnosis of ulcerative colitis (UC) and Crohn's disease (CD), the main subtypes of inflammatory bowel disease (IBD), has been challenging due to the constraints of the current techniques. N6-methyl adenosine (m6A) regulators have evolved as key players in IBD pathogenesis; however, their relation to its clinical setting is largely unexplored. This study investigated the potential of selected RNA methylation machinery and m6A target genes as serum biomarkers of UC and CD, their predictive and discriminating capabilities, and their correlations with laboratory data, interleukin (IL)-6, interferon-γ, disease activity scores, and pathological features. Fifty UC and 45 CD patients, along with 30 healthy volunteers were enlisted. The mRNA expression levels of the m6A writers methyltransferase-like 3 (METTL3) and Wilms-tumor associated protein (WTAP), and the reader YTH domain family, member 1 (YTHDF1), along with the m6A candidate genes sex determining region Y-box 2 (SOX2), hexokinase 2 (HK2), and ubiquitin-conjugating enzyme E2 L3 (UBE2L3) were upregulated in UC patients, whereas only METTL3, HK2, and UBE2L3 were upregulated in CD patients versus controls. Serum WTAP (AUC = 0.94, 95 %CI = 0.874-1.006) and HK2 (AUC = 0.911, 95 %CI = 0.843-0.980) expression levels showed excellent diagnostic accuracy for UC, METTL3 showed excellent diagnostic accuracy for CD (AUC = 0.91, 95 %CI = 0.828-0.992), meanwhile, WTAP showed excellent discriminative power between the two diseases (AUC = 0.91, 95 %CI = 0.849-0.979). Multivariate logistic analysis unveiled the association of METTL3 and UBE2L3 expression with the risk of CD and UC diagnosis, respectively, controlled by age and sex as confounders. Remarkable correlations were recorded between the gene expression of studied m6A regulators and targets in both diseases. Among UC patients, serum METTL3 and WTAP were correlated with UC extent/type, while WTAP was correlated with IL-6. Among CD patients, serum METTL3 and HK2 were correlated with CD activity index (CDAI) and CD location. In conclusion, m6A regulators and target genes are distinctly expressed in UC and CD clinical samples, correlate with disease activity and extent/location, and could serve as a novel approach to empower the diagnosis and stratification of IBD subtypes.

Characterization of Carbapenem-Resistant K. Pneumoniae Isolated from Intensive Care Units of Zagazig University Hospitals.

The advent of carbapenem-resistant Klebsiella pneumoniae (CRKP) poses a significant challenge to public health, as carbapenems are typically employed as a last resort to treat nosocomial infections caused by such organisms, especially in intensive care units (ICUs). This study aims to characterize the CRKP isolated from patients admitted to the Zagazig University Hospitals (ZUHs) ICU in Egypt. About 56.2%, 41.0%, and 32.4% of the isolates indicated the presence of blaNDM, blaOXA-48, and blaKPC, respectively. Carbapenemase-encoding genes were found in many isolates, and blaNDM was the most predominant gene. Nevertheless, this situation has become a heavy burden in developing countries, including Egypt, and is associated with substantial morbidity, mortality, and increased healthcare expenses.

Evaluation of different therapeutic approaches for spontaneous bacterial peritonitis.

BACKGROUND AND STUDY AIMS: Spontaneous bacterial peritonitis (SBP) is a significant cause of mortality in cirrhosis. Reducing toxic burden of infected ascitic fluid through paracentesis needs further studies as adjunctive therapy of SBP. We aimed to evaluate different therapies for SBP. PATIENTS AND METHODS: Thirty-six cirrhotic ascitic patients with SBP were examined and classified according to treatment modality (5-7 days) into: Group A received cefotaxime, group B received cefotaxime and albumin 1.5 g/kg body weight within 6h of SBP being diagnosed and 1g/kg body weight on day 3, group C received cefotaxime and paracentesis with volume dependent albumin infusion. Control group of 12 cirrhotic ascitic patients free from SBP were included. Routine laboratory tests, ascitic fluid analysis for leucocytes and culture were done, inflammatory mediators such as nitric oxide and tumour necrosis factor alpha were measured in serum and ascitic fluid. Duplex-Doppler assessment of portal flow volume and renal resistive index, Echocardiography to measure end diastolic and end systolic volumes, stroke volume and cardiac output were done. Tests were carried out before and after therapy. RESULTS: Treatment response was assessed by, cardiac haemodynamics, portal and renal flow and NO and TNF. All studied parameters; laboratory, cardiac, Doppler exhibited a significant improvement in group B in contrast to the other groups as demonstrated by post therapy reduction of (blood and ascitic fluid WBCs & PNLS, serum and ascitic NO & TNF and renal resistive index), elevation of (serum albumin and portal flow volume) and improvement of cardiac haemodynamic. CONCLUSION: Treatment of spontaneous bacterial peritonitis by cefotaxime and body weight based albumin infusion gave most favourable results compared to other regimens. Postulation of removing toxic burden through paracentesis has not been confirmed.

Association of adiponectin with cardiovascular events in diabetic and non-diabetic hemodialysis patients.

Adiponectin is a novel collagen-like protein synthesized by white adipose tissue. Its levels are decreased in obesity, type-2 diabetes and insulin-resistant states, and are increased in chronic renal failure. It has anti-inflammatory and anti-atherogenic properties. This study was planned to evaluate the levels of adiponectin in uremic patients with and without diabetes and to find any relationship between adiponectin levels and some cardiovascular risk factors, and to determine the possible predictive value of adiponectin for cardiovascular complications (CVC). The study included 100 subjects, 20 of them were healthy subjects and served as the control group (group I), 40 were uremic non-diabetic patients (group II) (half of them were without CVC, group IIA, and the other half were patients with CVC, group IIB) and, lastly, 40 uremic diabetic patients (group III) (half of them were without CVC, group IIIA, and the other half were patients with CVC, group IIIB). All subjects were subjected to complete clinical examination, including determination of mean arterial blood pressure (MABP), body mass index (BMI), waist to hip ratio, routine laboratory investigations, fasting plasma glucose, fasting plasma insulin, lipid profile (cholesterol, TG, LDL, HDL), determination of insulin resistance by homeostasis model assessment index (HOMA-IR) and estimation of serum levels of adiponectin. There was a significant increase in serum adiponectin levels in all the uremic patients (group II and group III) when compared with the control (group I) group, P <0.01; also, serum adiponectin levels were significantly decreased in uremic diabetic patients (group III) when compared with uremic non-diabetic patients (group II), P <0.01; but this was still higher than in the controls. The patients with CVC, whether uremic non-diabetic (group IIB) or uremic diabetic (group IIIB), had a significant decrease in serum adiponectin levels when compared with patients without CVC (group IIA and group IIIA), P <0.01. Serum adiponectin has a significant positive correlation with HDL and a significant negative correlation with MABP, BMI, plasma insulin, HOMA-IR, LDL, TG and cholesterol in all the patients. Therefore, it can be concluded that adiponectin levels in uremic patients, whether diabetic or non-diabetic, may be a good indicator of cardiovascular disease risk.

Soluble interleukine-2 receptor and MDR1 gene expression levels as inflammatory biomarkers for prediction of steroid response in children with nephrotic syndrome.

INTRODUCTION: Upregulation of interleukin-2 may be involved, not only in the pathophysiology of nephrotic syndrome, but also in steroid resistance treatment, by increasing expression of multidrug resistant gene-1 (MDR1) gene on lymphocytes and its product P-glycoprotein effluxing corticosteroid. Our aim was to assess the relation of serum soluble interleukin-2 receptor (sIL2R) levels and MDR1 gene expression on lymphocytes with nephrotic syndrome and its corticosteroids therapy. MATERIALS AND METHODS: We examined 40 children with nephrotic syndrome (15 cases of recent onset and 25 known cases with relapse) and 20 healthy children as a control group. We examined every patient twice at the time of disease activity and within 1 week of remission. RESULTS: A significant increase was found in sIL2R level and MDR1 gene in the patients in comparison with the control group whether in activity or remission, and they were significantly higher in activity than in remission. Levels of sIL2R and MDR1 gene expression in different subgroups were higher in known cases with relapse than in new onsets, both in activity and remission, and relatively higher in steroid-resistant than in steroid-sensitive ones. CONCLUSIONS: We propose sIL2R and MDR1gene expression levels as early predictors of steroid resistance in nephrotic syndrome for early control of disease by immediate introduction of cytotoxic drugs. This is the first report providing new insight into the use of sIL2R as a predictor of steroid resistance. Thus, wide-scale studies are needed to determine a cutoff level of sIL2R above which cytotoxic drugs are introduced.

Protein C levels in beta-thalassemia major patients in the east Nile delta of Egypt.

BACKGROUND AND OBJECTIVES: Thalassemic patients have an increased risk for thromboembolic complications. To determine if this might be due to a deficiency in protein C, we investigated the status of the protein C anticoagulant pathway in thalassemia major patients and its relationship to the hypercoagulable state. PATIENTS AND METHODS: Fifty patients with beta-thalassemia major (30 non-splenectomized and 20 splenectomized) and 20 healthy children as a control group were tested for levels of serum ferritin, liver enzymes, serum albumin, fibrinogen, protein C and protein S, thrombin antithrombin complex (TAT) and D-dimer. RESULTS: Thalassemic patients had lower levels of protein C and S and higher levels of D-dimer and TAT than the control group. These findings were more obvious in splenectomized patients and in those with infrequent blood transfusion. CONCLUSIONS: Protein C plays a major role in the hypercoagulable state in thalassemic patients. These findings raise the issue as to whether it would be cost-beneficial to recommend prophylactic antithrombotic therapy in high-risk thalassemic patients. A wider prospective study is necessary to delineate under which circumstances therapy might be needed, and at what level of protein C deficiency to start prophylactic antithrombotic therapy.

Impact of CD4+CD25high regulatory T-cells and FoxP3 expression in the peripheral blood of patients with systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is a systemic autoimmune disease characterized by various immunological abnormalities. Regulatory T cells (Tregs) CD4+CD25+ play an important role in maintaining tolerance to self-antigens controlling occurrence of autoimmune diseases. It has been shown that the transcription factor forkhead box P3 (FoxP3) is specifically expressed on CD4+CD25+T cells. FoxP3 has been described as the master control gene for the development and function of Tregs. A decrease in the number of CD4+CD25highFoxP3+ regulatory T cells can play a key role in the loss of tolerance to self antigens. The study was designed to assess expression levels of FoxP3 in peripheral CD4+CD25+ regulatory T cells in patients with SLE and to evaluate the level of some cytokines that are implicated in the extent of the disease activity. The study was carried out on 30 SLE patients, they were 27 females and 3 males, 10 age and sex matched healthy volunteers were studied as a control group. They were divided into two groups: group I: had active disease (12 patients) and group II: had inactive disease (18 patients) according to Systemic Lupus Erythematosus Disease Activity Index. All individual were subjected to CBC, ESR, s.creatinine, RF, CRP, C3, ANA, anti ds-DNA and flowcytometeric assay of CD4+CD25+ (Tregs) and FoxP3 for patients and controls. Quantitative determination of serum interleukin 10 (IL-10) and transforming growth factor-beta1 (TGF-beta1) concentrations in serum samples by ELISA technique. The results revealed a significant decrease of CD4+CD25high cells in peripheral blood in active lupus patients when compared with patients with inactive lupus and those in healthy controls. Intriguingly, the percentage level of FoxP3 on CD4+CD25high cells was significantly decreased in SLE patients with active disease (2.9 +/- 1.05) when compared with those with inactive SLE (3.5 +/- 0.8) and control groups (4.7 +/- 1.2) (P < 0.05). As regard cytokines levels; the level of IL-10 was significantly increased in patients with active and inactive disease (158.8 +/- 50.8, 82.8 +/- 14.08 respectively) when compared with the control group (P < 0.001). While, the level of TGF-beta1 was significantly decreased in patients with active and inactive disease (22.5 +/- 7.03, 29.07 +/- 10.14 respectively), when compared with the control group (P < 0.05). Our data revealed impaired production of Tregs in SLE patients, which may play a reciprocal role with some cytokines to affect the activity of the disease. Tregs cells should be the target to determine the clinical effectiveness of novel therapy to modulate Tregs in vivo besides the conventional treatments.