Aberrant cytokine and VCAM-1 expression in patients with viral and non-viral related liver cirrhosis.

The study aim was to compare the alterations in the expression levels of proinflammatory and chemotactic cytokines as tumor necrosis factor-alpha (TNF-α), interleukin (IL)-17A and IL-8, the down regulatory cytokine IL-10, in addition to the vascular cell adhesion molecule-1 (VCAM-1) gene in different groups of patients with cirrhosis due to various etiologies. This case-control study included 84 patients suffering from cirrhosis of viral and non-viral etiologies and 20 sex and age-matched healthy controls. All patients were subjected to detailed history taking, clinical examination, and liver function assessment. The expression levels of TNF-α, IL-17A, IL-8, IL-10, and VCAM-1 were assessed in peripheral blood mononuclear cells by real-time PCR. Patients with cirrhosis showed marked changes in the tested gene expression levels relative to the control group. Higher expression levels of all genes except IL-10 were seen in patients of the viral than in the non-viral groups. Most of the significant correlations of liver function parameters were observed with TNF-α in both the viral and non-viral groups, followed by IL-17A. Increased TNF-α and IL-17A presented potential risk factors for disease progression to cirrhosis of Child class C.

Regulatory T Cells and IL35 in Chronic Hepatitis C Related Cirrhosis and Hepatocellular Carcinoma.

Regulatory T cells (Tregs) play a fundamental role in maintaining immune homeostasis to balance between the tissue-damaging and protective effects of the immune response. There are strong evidences that Treg cells and their cytokines may play an important role in the induction of tolerance in the liver and progression of HCV infection. Herein, we investigated the frequency of Treg cells and interleukin 35 (IL-35) level in blood and their potential relationship to the various chronic hepatitis C (CHC) complications and outcomes. A total of 36 HCV infected patients subdivided into, CHC complicated with cirrhosis (HCV LC; n = 18), CHC complicated with hepatocellular carcinoma (HCV- HCC; n=18) and apparently healthy control group (n=18) were enrolled in this study. Treg cells percentages were determined by flow cytometric analysis and ELISA was used to measure IL35 serum levels. A significant increase in the frequency of peripheral Tregs and serum IL35 level was found in HCV HCC, and HCV LC groups compared with the control group. The frequency of peripheral Tregs and plasma (IL-35) levels were significantly positively correlated with viral load along with disease progression. We conclude that the higher percentage of Tregs and IL35 level in peripheral blood of HCV HCC and HCV LC groups compared to the control group may suggest their contribution to viral persistence and progression of HCV infection.

Concordance between peripheral and decidual NK cell subsets and killer immunoglobulin-like receptors in women with recurrent spontaneous miscarriages.

BACKGROUND: The role of decidual natural killer (dNK) cells in normal and complicated pregnancy and their relation with peripheral NK (pNK) cells remains unclear. The study aim was phenotypic analysis of pNK and dNK cells at time of miscarriage in recurrent spontaneous miscarriage (RSM) patients to assess whether measuring levels of pNK cell populations can reflect changes in dNK cells or not. METHODS: This study included 40 middle aged pregnant women in the 1st trimester subjected to evacuation because of a current miscarriage. They had a history of previous ≥ two unexplained miscarriages. Frequencies of pNK and dNK cells, based on the expression of CD56, CD16, inhibitory (CD158b) and activating (CD161) Killer immunoglobulin-like receptors (KIRs), were detected by flow cytometry. RESULTS: Percentages of CD56+ NK cells in peripheral blood and decidua were 17.5 % and 17.3 %, respectively. In both blood and decidua, CD56dim NK cells were exceeding CD56bright NK cells. The CD56dim CD16- NK cells were the predominating subset of NK cells, followed by CD56dim CD16dim. No substantial differences were detected in the levels of KIRs expression by the different NK subsets between blood and decidua. Abnormal up-regulation of both CD161 and CD158b on NK cells was observed in blood and decidua. CONCLUSION: At the time of miscarriage, patients with RSM have an extremely active immune system and an increased number of toxic NK cells both in blood and decidua. The pNK cells reflect dNK cell changes during miscarriage and may be a useful non-invasive predicting tool in reproductive failure setting.

Genetic Polymorphism and Serum Levels of RORc2 in Rheumatoid Arthritis.

RORc2 is the master transcription factor of T helper 17 cells. We aimed to evaluate whether RORc2 genetic polymorphism and serum levels have association with the risk and activity of rheumatoid arthritis (RA). RORC genetic polymorphisms were investigated by real time PCR. Serum RORc2 protein levels were determined by enzyme-linked immunosorbent assay. Protective effects of rs370515 CT, rs370515 CT + TT, rs3828057 CT, rs3828057 CT+TT and rs9826 GG genotypes were detected. The genotype-phenotype analysis showed no significant differences in the disease activity score 28 (DAS 28) under the recessive versus dominant genotypes. RORc2 protein serum levels were significantly higher in RA patients than controls (P= 0.001) and had a positive correlation with DAS-28. In conclusions, RORC genetic polymorphisms correlate with the risk but not activity of RA, whereas RORc2 serum levels have a positive correlation with both risk and activity of RA.

Occult Hepatitis B Infection Among Blood Donors in Al Azhar University Hospital, Upper Egypt: The Current Status After 25 years of Vaccine Introduction.

Occult hepatitis B virus infection (OBI) is a challenging topic with much debate regarding its clinical and virological relevance. The reliance on anti HBc seropositivity for diagnosis of OBI is still controversial. We aimed to determine the frequency of OBI among Egyptian blood donors, the role of anti HBc and HBe Ag as predictors of OBI and the possible risk factors. A total of 300 randomly selected hepatitis B surface antigen (HBsAg) negative blood donors at the Blood Bank of Al Azhar University Hospital in Assiut were included. Measurement of liver enzymes and screening for HBV core antibodies (anti HBc) and Hepatitis e antigen (HbeAg) were done. Quantitative HBV DNA measurement was achieved by real time polymerase chain reaction with a detection limit of 20 IU/mL after DNA extraction from the peripheral blood mononuclear cells (PBMC). The prevalence of OBI was 3.7% (11/300) among the donors and the majority had low level viremia (63.6%). Anti HBc was detected in 20.7% (62/300) (group I). OBI was detected in 14.5% (9/62) in group I compared to 0.84% (2/238) in the anti-HBc Ab negative donors (group II). Anti HBc had a sensitivity and specificity of nearly 82% for detection of OBI with a high negative predictive value (99.16%). HBe Ag was detected in only 1.6% (1/62) of group I. There were no statistical significant differences regarding the liver enzymes, demographic data or risk factors between group I and II and even between cases of OBI and those without. We conclude that OBI exists in an alarming percentage among Egyptian blood donors. Anti HBc should be introduced in the routine blood screening. Negative anti HBc results ensures safe blood, while positive results need nucleic acid confirmation especially if given to high risk recipients. More in-depth evaluation of the immunization program is needed.

Role of T-Helper 9 Cells in Chronic Hepatitis C-Infected Patients.

Hepatitis C virus is a hepatotropic virus that is transmitted parenterally. Viral infections are usually associated with modulations of the immune cells, leading to enhanced viral survival and spreading, and accordingly, life-threatening complications. Recently, it has been proposed that a new subset of T-helper, named T-helper 9, is involved in the pathogenesis of different immunopathological conditions, such as allergies, tumors, and viral infections. Some studies reported a protective role, and others described a pathogenic potential for the T-helper 9 cells. Here, we present evidence that T-helper 9 cells are dynamically increased with increasing the pathogenic strategy for hepatitis C virus (HCV). Furthermore, viral clearance is associated with a decrease in T-helper 9. The increase in T-helper 9 was paralleled with an increase in its receptor expression. Taken together, our data suggest that T-helper 9 cells play an important role in the pathogenesis of HCV, and is directly associated with HCV-related complications.

Regulatory T cell subsets in children with systemic lupus erythematosus.

The aim of this work was to quantify CD4(+)CD25(+)Foxp3(+) T cells (Tregs) in Egyptian children with SLE and to correlate these findings with their disease activity scores and drug therapy. We enrolled 37 Egyptian children with active SLE. Disease activity was assessed by measuring serum levels of anti-dsDNA antibody and by the SLEDAI scores. Twenty healthy children were also enrolled as normal controls. The CD4+CD25+, CD4+CD25(bright), and CD4+CD25(dim) cells in patients were significantly increased in comparison to controls. There was no significant difference in the Foxp3 gated on CD4+CD25(bright) and CD4+CD25(dim), but there was a significant increase when gated on CD4+CD25- and whole CD4+ cells in patients than controls. There was no significant difference among patients with different degrees of activity on different lines of treatments and their outcomes as regards all studied values. There was no significant correlation between SLEDAI score and any of the studied parameters except for a significant negative correlation with gated lymphocytes. There is increased expression of Foxp3 in CD4+ T cells mostly CD25- in Egyptian children with active SLE under corticosteroid treatment regardless of disease activity.

Comparison of different phenotypic and molecular methods for detection of methicillin-resistant Staphylococcus aureus in intensive care patients.

Detection of methicillin-resistant Staphylococcus aureus (MRSA) has been problematic ever since its discovery. This work was performed to evaluate the effectiveness of different phenotypic methods for MRSA detection in intensive care patients. Three hundred and eighty-nine specimens from 100 patients were inoculated onto mannitol salt agar. All S. aureus isolates were examined by four selective culture media [ORSAB, MSO, MSA-Cefox, OAS], two disc diffusion methods [CDD and ODD], and MicroScan panel for MRSA. Polymerase chain reaction (PCR) for mecA gene was performed as the gold standard. S. aureus isolates were revealed from 56 patients, 41 of them were found to be MRSA by PCR. CDD yielded the best sensitivity (97.6%), followed by ODD and MSA-Cefox (92.7%). CDD, MSA-Cefox and OAS showed the best specificity (100%). In conclusion, MSA-Cefox and CDD showed improved sensitivity and excellent specificity compared to other methods. It is advisable to use the two methods for MRSA detection if PCR is not available.

Dual effect of nitric oxide donor on adjuvant arthritis.

The effect of medical use of NO donors on the pathogenesis of arthritis is still yet unclear. We investigated the effects of the NO donor, sodium nitroprusside (SNP), on the pathogenesis of adjuvant-induced arthritis in rats. Rats were given SNP intraperitoneally either from day 5 to day 14 (as a prophylactic protocol) or from day 16 to day 25 (as a therapeutic protocol) after inoculation of adjuvant. SNP administration, whether prophylactic or therapeutic, in doses of 0.1 and 1 mg/kg/d significantly aggravated pathogenesis of adjuvant arthritis in rats. SNP-treated rats showed significant (P<0.05) increase in arthritis index, hind paw volume, ankle joint diameter and hyperalgesia compared with control adjuvant arthritic rats. However, in adjuvant rats given the smallest dose of SNP (0.01 mg/kg/d), arthritis index, volume of hind paws, ankle joint diameter, body weight loss, and hyperalgesia were significantly lower than that of control adjuvant rats. After 30 d of the induction of adjuvant arthritis, TNF alpha levels exhibited insignificant changes either in control adjuvant rats or in rats given SNP compared with control non adjuvant rats. IL-10 levels in adjuvant control rats and adjuvant rats given 1 mg or 0.1 mg/kg/d from day 15 to day 25 were significantly lower than that of control non adjuvant rats. Histopathology examination of ankle joint showed that large doses of SNP (1 mg or 0.1 mg/kg/d) increased the mononuclear cells infiltration and erosion of cartilage induced by adjuvant while the infiltration of the inflammatory cells in the synovium of adjuvant rats treated with 0.01 mg/kg/d was minimal and the pannus was inhibited with alleviation of erosion of articular cartilage. Prophylactic small dose of SNP improved the histological status more than the therapeutic small dose. The present work reveals that SNP administration, either prophylactic or therapeutic, was deleterious in higher doses. However, the smallest dose used 0.01 mg/kg/d attenuates joint inflammation, hyperalgesia and body weight loss in adjuvant arthritic rats. These results suggest that small dose of NO donor may exert partial protective effects while the safety of the clinical use of NO donors, in higher doses, in patients with rheumatoid arthritis is questioned.

Oxidative stress as a common mediator for apoptosis induced-cardiac damage in diabetic rats.

AIM: To investigate the possible role of oxidative stress as a common mediator of apoptosis and cardiac damage in diabetes. MATERIALS AND METHODS: This experimental work was conducted on 5 groups of Wistar rats. Group I was the control group. Diabetes type 1 was induced in other groups (by streptozotocin) and animals received insulin or vitamin E (300 mg /kg body weight), both insulin and vitamin E, or no treatment for 4 weeks according to their group. At the end of the study, serum and cardiac tissues were examined for biochemical parameters of cardiac function, oxidative stress and apoptosis. Electron microscopy pictures of cardiac tissue were also evaluated for signs of cardiac damage RESULTS: Markers of oxidative stress, apoptosis, inflammation as well as manifestations of cardiac damage as assessed by electron microscopy were significantly decreased in rats treated with both insulin and vitamin E when compared with untreated diabetic rats or rats treated with either insulin or vitamin E alone CONCLUSION: Administration of both vitamin E and insulin was effective in reducing markers of oxidative stress and apoptosis and improving parameters of cardiac function in experiments animals. Antioxidants might prove beneficial as an adjuvant treatment in addition to insulin in type 1 diabetes associated with manifestations of cardiac complications.