Role of broad-range polymerase chain reaction in diagnosis of infective endocarditis

Positive blood culture is one of the modified Duke's major criteria for diagnosis of infective endocarditis [IE]. Nevertheless, negative blood culture may occur in IE due to prior antimicrobial utilization or infection with fastidious microorganisms. Direct amplification of bacterial DNA by using broad-range PCR primers provides an alternative approach to the detection of pathogens from clinical specimens. Amplification of the 16S and 18S ribosomal RNA gene in DNA extracted from excised heart valve tissue [HV] has been used to provide etiological diagnosis in patients with culture-negative IE. In this study we aimed to assess the usefulness and the practicality of broad-range PCR [brPCR] in diagnosis of infective endocarditis for the first time in Endocarditis Service, Cairo University and for the first time in Egypt. HV surgically removed from 32 patients with definite IE were included in the study. H V were subjected to brPCR and culture. By brPCR, 20 [62.5%] valves were positive for bacteria, of these seven [35%] valves from blood culture positive patients and 13 [65%] of culture-negative patients. Eleven [34.4%] of all patients were positive for fungus by broad-range PCR, all from patients with negative blood culture for fungus. DNA amplification with universal primers is a promising diagnostic tool in infective endocarditis patients where routine laboratory culture failed to identify the pathogen and diagnosis in the coming decade will likely rely more on PCR-based methods

Vascular endothelial growth factor and soluble adhesion molecules as a diagnostic markers for spontaneous bacterial peritonitis in cirrhotic liver disease

Spontaneous bacterial peritonitis [SBP] is a frequent and severe complication in cirrhotic patients with ascites that usually results in renal failure and death despite the efficacy of the current antibiotic therapy. The aim of this study was determine serum and ascitic fluid of soluble-L selectin [s-L Selectin], intracellular adhesion molecule-1 [ICAM-1], Vascular cell adhesion molecule-1 [VCAM-1] and vascular endothelial growth factor [VEGF] in cirrhotic patients, and to search for a relationship between them and SBP. This study was performed on 30 cirrhotic patients with SBP. Their ages ranged [from 38-55 years] with mean of [32 +/- 5.5], 30 cirrhotic patients with non-infected ascites; their ages ranged [from 30-52 years] with mean of [35 +/- 6.5]. This group considered as cirrhotic control group and 20 healthy control subjects their ages ranged [from 28-55 years] with mean of [30 +/- 7.5]. Serum and ascitic fluid of adhesion molecules as well as VEGF levels were significantly higher in cirrhotic patients with SBP as well as cirrhotic patients with non-infected ascites as compared to healthy control group. There were significant increase in serum and ascitic fluid level of leukocyte, PMN and ICAM-1 in SBP as compared to cirrhotic with non-infected ascites. There was non-significant decrease in serum and AF level of VEGF in cirrhotic control group as compared to SBP group. The ascitic fluid PMN and s-L Selectin were higher in culture positive SBP patients particularly in those with gram positive isolates, where these are non-significant increase in serum and ascitic fluid level of VEGF in culture positive SBP than culture negative cases. Positive correlation was found between serum and ascitic fluid level of ICAM-1 in SBP and non-infected cirrhotic group. Also, positive correlation was found between VEGF levels in serum ascetic fluid levels in both cirrhotic groups [SBP and non-infected cirrhotic group]. These data suggest that: Significant elevated level of VEGF in both SBP and non infected cirrhotic patient may have pathophsiological consequences of local regulation of vascular tone and endothelial permeability, significant elevated level of adhesion molecules in both SBP and non-infected cirrhotic patients are due to inflammatory response and endothelial cell activation. Serum and ascetic fluid of ICAM-1 can be used as useful marker for diagnosis of SBP and for monitoring the treatment of cirrhotic patients

Leptin level and plasma prothrombotic factors in obese subjects

Obesity is associated with increased cardiovascular morbidity and mortality. Abnormalities in coagulation and haemostasis represent a well-known link between obesity and thrombosis [both arterial and venous]. In human, production of a depocyte-derived peptide, leptin has been linked to adiposity; recent studies have shown that plasminogen activator inhibitor-1 [PAI-1], a prothrombotic factor associated with atherosclerosis complication is also produced in adipose tissue. Several studies reported that obese subjects had elevated levels of vonWillbrand factor [vWF] and fibrinogen. The aim of this work is to examine the relationship of obesity, fat distribution and serum leptin concentration with plasma levels of Prothrombotic factors [PAI-1 - [vWF] and fibrinogen] and lipid pattern [Triglycerides and Cholesterol] as metabolic parameters. The body fat distribution was evaluated by measuring the body mass index [BMI] and waist-to-hip ratio [WHR]

pathogenesis of cytokines in preportal fibrosis of human infected with schistosomiasis and viral hepatitis

Cytokines are polypeptides exhibiting a variety of biological activities including metabolic, inflammatory, hematopoietic and immunologic properties. They play an important role in the pathogenesis of various diseases. Inflammation is commonly observed in liver diseases and is frequently complicated by fibrosis and cirrhosis in end-stage disease. The only curative treatment for cirrhotic patients is liver transplantation. Cytokines play a key role in the regulation of immune responses. In viral hepatitis the production of inappropriate cytokine level appears to contribute to viral persistence and to affect response to therapy. The aim of this study is to investigate the level of endogenous IL-1B, IL-6 and IL-10 to determine their relation with liver fibrosis. Forty patients with chronic liver disease and 10 normal adults as control group were studied. Patients in this study were classified into four groups according to etiology of chronic liver disease: Group I [10 patients with bilharzia liver disease],Group II [10 patients with chronic hepatitis C], Group III[10 patients with chronic hepatitis B]and Group IV [10 patients with chronic hepatitis B and C]. All patients with chronic liver disease [n=40] showed highly significant elevation of serum IL-1B, IL-6, IL-10 mean +/- SD were [106.4 +/- 47.8] [P<0.01] [26.3 +/- 11.1] [P<0.01] [135.4 +/- 73.9] [P<0.01] respectively when compared to control group. After classifying the patients into 4 groups each group showed highly significant elevation of serum IL-1B, serum IL-6 and serum IL-10 in each group when compared to control group[p < 5051]. Regression analysis showed negative significant correlation between serum IL-10 and IL1B [r=-0.64, P<0.05], highly negative significant correlation between IL-10 and IL-6 [r=-0.72, P<0.01] in all patients with chronic liver diseases, also there was highly significant positive correlation between serum IL-1B and serum IL-6 [r=0.83, P<0.01]. Ten patients with bilharzia liver disease [group I] showed highly negative significant correlation between serum IL-10 and each of serum IL-1B and serum IL-6 [r=-0.9, P<0.01] [r=0.8, P<0.01] respectively, and there was highly significant positive correlation between serum IL-1B and serum IL-6 [r=0.96, P<0.01] .There was significant correlation between prothrombin concentration and each of serum IL-10, serum IL-1B and IL-6 [r=0.7, P<0.05], [r=0.68, P<0.05], [r=0.74, P<0.05] respectively. Ten patients with chronic hepatitis C virus [group II] also showed highly negative significant correlation between serum IL-10 and each of serum IL-1B and serum IL-6 [r=-0.9, P<0.01] [r=-0.9, P<0.01] respectively. There was highly significant positive correlation between serum IL-1B and serum IL-6 [r=0.83, P<0.01] and significant correlation between serum IL-1B and serum ALT[r=0.63, P<0.05]. As regard [group III] patients with chronic hepatitis B virus there was negative significant correlation between serum IL-10 and IL-1B [r=-0.63, P<0.05], but no significant correlation between serum IL-10 and serum IL-6 and there was highly positive correlation between serum IL-1B and serum IL-6 [r=0.90, P<0.01]. Ten patients with chronic hepatitis C and B virus [group VI] showed highly negative significant correlation between serum IL-10 and each of serum IL-1B and serum IL-6 [r=-0.82, P<0.01] [r=-0.80, P<0.01] respectively. There was highly significant positive correlation between serum IL-1B and serum IL-6 [r=0.88, P<0.01] and significant correlation between serum IL-1B and serum ALT[r=0.63, P<0.05], and prothrombin concentration [r=0.67, P<0.05]. A significant correlation between the level of serum IL-1B, IL-6, and serum IL-10 and degree of fibrosis was found. The increase in serum level of IL-1B, IL-6 was associated with increase the degree of fibrosis but the mild and moderate fibrosis were associated with higher level of IL-10 while patients with marked degree of fibrosis were associated with lower level of IL-10

Interleukin-10, Interleukin- 16 and Interferon-gamma in serum of patients with rheumatoid arthritis and correlation with disease activity

Rheumatoid arthritis [RA] is a systemic inflammatory disease of unknown etiology. The rheumatoid synovium is characterized by infiltration of T cells, macrophage, B cells, and proliferating fibroblasts which aggressively invade cartilage and bone, thus destroying joints' ability to function. In rheumatoid arthritis [RA] both an imbalance between excessive production of the proinflamatory and ant-inflammatory cytokines and skewing of the T cell to a T helper like response. Cytokines have been shown to play a modulatory role in the pathogenesis of RA. The imunoregulatory cytokine IL-10 increases autoantibody production by B cell stimulates its survival, proliferation and differentiation. Moreover IL-10 inhibits the generation of proinflamatory cytokines and proliferation of T helper lymphocyte. Interleukin 16 might play a role in the pathogenesis of chronic inflammation in RA. It has a proinflamatory properties by promoting recruitment of T cells into the rheumatoid synovium. Also IFN-gamma is of interest because of the role it plays in the initiation and perpetuation of T helper cell. Serum level of IL-10, IL-16, and IFN-gamma were determined in patients with rheumatoid arthritis in relation to disease activity. All patients with RA [n=30] showed highly significant increase in ESR, CRP, IL-10, IL16, IFNgamma compared to control group [p<0.01]. Positive correlation were found between IL-10 and each IL-16 and IFN-gamma [p<0.001] [r=0.63, 0.55] respectively, and highly significant correlation between IL-16 and IFNgamma [p<0.001] [r=0.89]were determined. Results showed positive correlation between ESR and each IL-10, IL-16, IFNgamma [p<0.001] [r=0.67, 0.87, 0.75] respectively. And highly significant correlation between CRP and each IL10, IL-16, IFNgamma [p<0.001] [r=0.0.71, 0.83, 0.73] which indicate relation between increase level of cytokine with disease activity. These data suggest that there is increased production of IL-10, IL-16, and IFN-gamma in patients with rheumatoid arthritis, and that it is correlated with the disease activity. These cytokines are interesting for further research and novel therapeutic approach in this inflammatory disease