Circulating cell free DNA as a predictor of systemic lupus erythematosus severity and monitoring of therapy

Background: Systemic lupus erythematosus [SLE] is the most heterogeneous chronic autoimmune disease; it is characterized by the presence of auto reactive B and T cells, responsible for the aberrant production of a broad and heterogeneous group of autoantibodies. Recent studies using various detection methods have demonstrated the elevations of circulating DNA in SLE patients

Aim of the study: The current study aimed to measure cell-free DNA [cf-DNA] in SLE patients as a potential tool to predict disease activity and treatment follow up

Subjects and methods: 52 of SLE patients with age ranging from 10 to 48 years were randomly selected and 25 healthy subjects with age and gender matched with the patients were included as a control group. Thorough clinical examination stressing on the central nervous system, vascular, renal, rash, musculoskeletal, mucocutaneous manifestations, and fever was done for patients. The following investigations were done: Complete blood count [CBC], kidney function tests, C-reactive protein [CRP], routine autoantibodies for autoimmune diseases, complements [C3 and C4], anti-nucleosome antibodies and cf-DNA by real time PCR [RT-PCR]

Results: The levels of anti-double stranded DNA [anti-dsDNA], anti-nucleosome Ab, and cf-DNA were significantly increased in SLE patients compared to controls. The cf-DNA level was correlated to markers of disease severity namely CRP and anti-nucleosome. A significant reduction in levels of cf-DNA, anti-nucleosome Ab and anti-dsDNA was noticed after therapy

Conclusion: Our findings support that the measurement of cf-DNA appears to be a useful marker in addition to laboratory tests used in SLE diagnosis. High correlation with markers of disease severity suggesting its role in disease pathogenesis and decreasing its level after therapy makes it to be a marker of treatment follow-up

diagnostic and prognostic significance of circulating and intrahepatic endoglin in liver fibrosis associated HCV infection

Endoglin [CD 105], a transforming growth factor-beta [TGF-beta 1] co-receptor, has been implicated in liver fibrogenesis and carcinogenesis. To determine CD 105 in liver tissue and serum of patients with chronic hepatitis C virus [HCV] infection and to correlate CD 105 expression to stage of fibrosis and serum markers of liver fibrosis; TGF-beta 1 and hyaluronic acid [HA]. Liver biopsies from 42 chronic HCV patients and 20 healthy subjects [donors for liver transplantation] were used for tissue quantification of CD 105 by ELISA and Western blot analysis then correlating findings with tissue TGF-beta 1. Serum CD 105 and HA from patients and control subjects were determined by ELISA. According to the stage of liver fibrosis assessed by Ishak scoring system, patients were classified into 3 groups: Group I: with mild fibrosis [stage 1-2] included 16 patients [11 males, 5 females, mean age 34.5 +/- 7.2 years], Group II: with moderate fibrosis [stage 3-4] included 15 patients [12 males, 3 females, mean age 36.5 +/- 4.1 years] and Group III: with severe fibrosis [stage 5-6] included 11 patients [8 males, 3 females, mean age 41.5 +/- 5.6 years]. Patients with severe fibrosis revealed significantly higher intrahepatic expression of CD 105 and TGF-13 1 than those patients with moderate fibrosis [p<0.05, p<0.001], mild fibrosis [p<0.01, p<0.001] and normal liver [p<0.001, p<0.001]. Serum levels of CDI 05 were 7.9 +/- 1.4 ng/mL, 5.4 +/- 0.54 ng/mL and 3.6 +/- 0.52 ng/mL for patients with severe, moderate and mild fibrosis respectively with a significant difference [p<0.001]. Interestingly, serum CD1O5 showed a positive correlation with severity of fibrosis and serum HA concentrations. At a cut off 3.12 ng/mL, CD 105 gave a sensitivity of 70%, specificity of 89% with area under curve [AUC] of 0.781 for diagnosis of liver fibrosis and at cut off 43.2 ng/mL. HA had a sensitivity of 80%, specificity of 96% and AUC of 0.903. Data from this study provide evidence for a new clinical application of endoglin [CD 105] as a complementary biomarker in diagnosis and prognosis of liver fibrosis as well as a possible therapeutic role of endoglin antagonist in the management of cirrhosis

Epstein barr virus DNA in hepatocelullar carcinoma tissues from hepatitis C positive Egyptian patients

Hepatocellular carcinoma [HCC] is one of the most frequent malignant tumors. It possesses the characteristics of high malignancy, rapid progress and poor prognosis. In recent years, studies have suggested that Epstein-Barr virus [EBV] is associated with HCC although opposite results have been subsequently reported. The present study was to determine the prevalence of EBV in HCC Egyptian patients, and whether EBV acts synergistically with hepatitis viruses in HCC carcinogenesis. The study included 61 patients, 20 HCV positive patients without HCC [16 males and 4 females] and 41 patients with proved HCC. They were subclassified into 3 groups [21 HCV positive [18 males and 3 female], 10 HBV positive [8 males and 2 females] and 10 HCC patients negative for both HCV and HBV [7 males and 3 females]]. Thorough clinical examination, abdominal ultrasonography and liver spiral CT were done. Liver function tests and serum alpha-fetoprotein [AFP], viral hepatitis markers for B and C, anti-EBV early antigen [EA-IgM], virus capsular antigen [VCA-IgMl and HCV RNA by reverse transcription PCR [RT-PCR] were measured. EBV-BamHI W DNA, and EBV-LMP1 DNA were performed by conventional PCR in the tumorus liver tissue of 41 HCC patients and the 20 noncarcinoma patients [HCV without HCC]. The positive ratios were compared between HCC subgroups and non tumorus HCV group. Our results revealed that, EBV-BamHI W DNA and/or EBV-LMP1 DNA were positive in 25 [40.9%] among overall 61 studied cases. In HCC patients, EBV-BamHI W DNA and/or EBV-LMP1 DNA were positive in 13 [61.9%] out of 21 HCV positive, 2 [20%] out of 10 HBV positive cases, 3 [30%] out of 10 cases negative for both HCV and HBV. However, it was positive in 7 [3 5%] out of 20 HCV cases without HCC [non tumotus cases]. The rate of EBV infection in HCC with HCV positive cases was significantly higher [Fisher exact=4.6 1; p<0.05] than HCC with HBV positive ones, HCC cases negative for both B and C virus [Fisher exact-4.28; p<0.05] and chronic HCV [non tumours] cases [Fisher exact=4. 19; p<0.05]. In addition, HCC in EBV DNA positive cases was associated with high HCV viral load, AST, ALT, low serum albumin, while there was no relation to AFP serum levels. In conclusion: the existence of EBV infection in HCC tissues suggests that EBV may be involved in the hepatocellular carcinogenesis in Egypt

Relationship between prothrombin fragment 1+2 and helicobacter pylori associated chronic gastritis

Helicohacter pylori [H. pylori] have been suggested to be an independent risk factor for cardiovascular and cerebral stroke; through the activation of clotting system following chronic inflammation. Among proinflammatory cytokines, tumour necrosis factor-alpha [TNF-alpha] and interleukin-8 [TL-8] which have been previously demonstrated to activate the clotting system in human. This study aimed to investigate: [i] if there is a difference between the circulating levels of prothromhin fragment 1+2 [F1+2]; a sensitive marker of thrombin generation in patients with chronic gastritis according to N. pylori status, [ii] the relationship between circulating and mucosal levels of TNF-alpha, IL-8 and Fl+2 and [iii] If H. pylori eradication modifies the level of these cytokins and clotting system activation. Forty-two patients with chronic gastritis were diagnosed with upper endoscopy, and enrolled in this study, divided into 2 groups on the basis of histopathological examination of gastric antral biopsies and culture. Group I: included 28 H. pylori positive patients, and group H: 14 H. pyIori negative patients chronic gastnitis. A triple eradication therapy for H. pylori [omeprazole, amoxacillin and tinidazole] was given for the 28 H. pylori positive patients [Group I] and H. pylori eradication was evaluated after 2 months. Gastric mucosal levels of TNF-alpha and IL-8 as well as plasma levels of TNF-alpha, IL-8 and F1+2 were measured in group II and before and 2 months after H. pylori eradication in group I. Our results showed that: patients with H pylori positive gastritis had a significant increase in the mean value of plasma F1+2 mucosal and plasma levels of TNF-alpha and IL-8 compared with group II [P<0.001]. There was a significant positive correlation between plasma level of F1+2 and plasma levels of TNF-alpha and IL-8 [r=0.89, p<0.00, r=0.88, p<0.000] respectively and between the plasma levels of F1+2 and gastric mucosal levels of TNF-alpha and IL-S [r=0.89, P<0.000; r=0.86 P<0.001] respectively. There was a significant reduction in the mean value of plasma levels of TNF-alpha. IL-8 and F1+2 as well as fibrinogen 2 months later after H. pylori eradication. While in-group II [H pylori negative chronic gastrius] there was no significant correlation between the studied parameters. In conclusion, the present study showed a close relationship between plasma levels of prothrombin F1+2, and both plasma and gastric mucosal values of TNF-alpha and IL-8 in cases with H. pylori associated chronic gastritis. These findings suggest that H. pylori may represent a trigger factor for clotting system activation through persistent inflammatory stimulation, cytokines production and consequently H. pylori eradication may reduce the risk of hyper-coagulability state or thrombotic events which may be associated in these patients

Total antioxidant status and erythrocyte superoxide dismutase and glutathione peroxidase in rheumatoid arthritis patients

This study included 54 patients with rheumatoid arthritis [RA], 20 patients with osteoarthritis [OA] not receiving medication and 26 apparently healthy subjects as a control group. All groups were tested for total antioxidant status, erythrocyte superoxide dismutase and glutathione peroxidase levels, where the active rheumatoid arthritis group showed decreased levels of these antioxidants compared with the other groups. The results of the present study confirmed the supposition of an increased oxidative stress in the active rheumatoid arthritis patients and low erythrocyte antioxidant levels in terms of superoxide dismutase, glutathione peroxidase activities and plasma total antioxidant status and their association with an increased rheumatic activity. The study recommended the usage of antioxidant drugs as therapeutic agents. Also, the monitoring of total antioxidant status, erythrocyte superoxide dismutase and glutathione peroxidase levels might be helpful when assessing the therapeutic efficacy of anti-rheumatic drugs