Ethnic variation of IL-4 intron 3 VNTR gene polymorphism; its association with type 2 diabetes mellitus and its complication (neuropathy) in Egyptian subjects.

Type 2 diabetes mellitus (T2DM) has multigenetic and environmental interactive factors. Although diabetic neuropathies (DPN) are the most common, but at the same time, the least recognized and understood long-term complication of diabetes. This study aimed to investigate the association of IL-4 VNTR gene polymorphism with T2DM complicated with neuropathy in Egyptian subjects. This is a case control study including 102 T2DM Egyptian patients, plus 188 unrelated healthy individuals as controls. They were evaluated for variable number tandem repeat (VNTR); 70 base pair repeats located in the intron 3; of IL-4 gene using the PCR technique. Homozygote frequency of the three-repeat allele (A1/A1) genotype of IL-4 VNTR was nearly equal among diabetic cases and controls (60.8% vs. 62.2%, respectively). Heterozygous frequency of (A1/A2) genotype was higher among controls compared to cases (33.5% vs. 19.6%, respectively) but not statistically significant. The (A2) allele had a significantly higher frequency in diabetic cases compared to controls (29.3% vs. 21.0%, respectively) while the (A1) allele had lower frequency but not significant one (70.7% vs. 79.0%, respectively). Comparing cases complicated with diabetic neuropathy vs. noncomplicated cases regarding their polymorphic IL-4 (VNTR) genotypes revealed a nonsignificant lower frequency of (A1A1) genotype (57.1% vs. 65.1%, respectively, p = .57) with a higher combined (A2A2 + A1/A2) genotype frequency (42.9% vs. 34.9%, respectively). Only two haplotypes (A1) & (A2) of IL-4 (VNTR) gene were recognized among Egyptian population; (A2) allele may influence in diabetes but not its complication (neuropathy) among Egyptian diabetic patients.

Enhancing ex vivo expansion of cord blood-derived unrestricted somatic stem cells for clinical applications.

OBJECTIVES: To study effects of serum-containing medium (SCM) versus serum-free medium (SFM) and influence of seeding density, on rate of expansion of cord blood (CB) unrestricted somatic stem cells (USSCs), as a prerequisite for evaluating their therapeutic potential in ongoing clinical trials. MATERIAL AND METHODS: Isolation, propagation and characterization of USSCs from CB samples were performed and followed by their passage 3 culture in SCM and SFM, at cell densities of 5, 50, 500 and 5000 cells/cm(2) . RESULTS: The cells were CD44(+) , CD90(+) , CD73(+) , CD105(+) , CD34(-) , CD45(-) , and HLA-DR, with Oct4 & Sox2 gene expression; they were differentiated into osteoblasts and adipocytes. USSCs cultured in SCM had significantly higher population doubling levels (P < 0.01) than those cultured in SFM. Those cultured in SCM at 5 cells/cm(2) and those cultured in SFM at 50 cells/cm(2) had significantly higher population doubling (P < 0.01) levels than those cultured at higher cell densities. CONCLUSIONS: For scaling up of USSCs from 106 (?) to 1012 (?) in 6 weeks, culturing of CB-derived cells of early passage (≤P3) in SCM at low cell seeding density (5 cells/cm(2) ) is suggested for increasing cell count with lower passaging frequency, followed by culture of expanded USSCs at 50 cells/cm(2) in SFM, to avoid undesirable effects of bovine serum in clinical applications.

Serum IL-10 and IL-6 levels at diagnosis as independent predictors of outcome in non-Hodgkin's lymphoma.

Interleukin 10 (IL-10) and interleukin 6 (IL-6) are widely investigated in solid tumors as being important prognostic factors. IL-10 and IL-6 serum levels were measured by enzyme-linked immunosorbent assay from sera taken from 40 non-Hodgkin's lymphoma (NHL) patients before and after treatment and from 20 healthy controls. The patients had been observed for at least 18 months or until death. IL-10 and IL-6 were significantly higher in NHL patients compared to controls. IL-6 was correlated with IL-10 (r = 0.451) and with B symptoms (weight loss > 10% during the last 6 months, unexplained fever and night sweats) (r = 0.447). IL-10 and IL-6 were significantly higher in non survival compared to survival group. High pretreatment IL-10 and IL-6 was associated with poor overall survival. These results show that IL-10 and IL-6 levels are elevated in NHL patients and seem to suggest that simultaneous elevation of IL-10 and IL-6 is a powerful negative prognostic parameter in NHL.

Effect of the broad spectrum anthelmintic drug flubendazole upon Schistosoma mansoni experimentally infected mice.

The effect of the broad spectrum anthelmintic drug flubendazole (methyl 5-(p-fluorobenzoyl)-2-benzimidazolecarbamate, CAS 31430-15-6), a mebendazole derivative with a molecular weight of 313.29, on Schistosoma mansoni infection in mice was evaluated. Moreover, the relationship between the posttreatment worm burden, hepatic granuloma volume, and serum immunoglobulin profile (immunoglobulin G and immunoglobulin M, IgG and IgM), was also investigated. Two main groups of Swiss albino mice infected with Schistosoma mansoni cercariae were used in the experiment. Group I consisted of infected untreated control mice. The mice of group II were submitted to treatment with flubendazole 100 mg/kg body weight as single oral dose at different time intervals: Group IIa received treatment 24 h before infection. Group IIb received treatment 4 h after infection. Group IIc received treatment 25 days after infection. Mice treated 25 days after infection, compared to those treated in other time intervals, revealed a significant reduction in the recovery of adult schistosomes after portal perfusion (79.5%), a lower immunoglobulin level (IgG and IgM), and the smallest granuloma mean diameter (220.0 +/- 10.3 microns). These data were less salient in mice treated 4 h after, and 24 h before infection.

Induction of hepatic egg granuloma hyporesponsiveness in murine schistosomiasis mansoni by intravenous injection of small doses of soluble egg antigen.

This work was designed to test whether hyporesponsiveness to schistosomal egg antigen (SEA) was associated with reduction in size of hepatic granulomas. Multiple small doses of SEA (10 micrograms x 4) were injected intravenously (i.v.) into C57B1/6 mice either at 7 or 30 days prior to cercarial exposure. Eight weeks postinfection, hepatic histopathology and granuloma diameter were studied. SEA-induced lympho-proliferative response, splenic cytokines (IL-2, IL-4 and IL-5) and serum antischistosomal IgG were assessed. Worm burden and tissue egg load were counted. Compared to infected controls, the SEA-treated groups showed decrease in granuloma diameter, remarkable increase in the percentage of degenerated ova within hepatic granulomas and amelioration of histopathological changes. SEA lymphoproliferative response, and levels of Il-2 and IL-4, were lower in SEA-treated groups than infected controls. The levels of IL-5 and antishistosomal IgG were comparable to the infected controls. The intensity of infection was not influenced by i.v. injection of SEA. The present data show that i.v. administration of multiple small doses of SEA induced granulomatous hyporesponsiveness with amelioration of hepatic pathology and acceleration of egg destruction.

Effect of early treatment with praziquantel on serum connective tissue metabolite markers in children and adolescents with intestinal schistosomiasis mansoni.

This work was designed to assess the reflection of early treatment by praziquantel (CAS 55268-74-1, EMBAY 8440, Biltricide) on serum connective tissue metabolite markers (hyaluronic acid and procollagen III peptide) in patients with active intestinal schistosomiasis. Children and adolescent subjects from primary and secondary schools in an endemic area of schistosomiasis mansoni were included. Age-matched subjects from an urban area served as normal controls. All subjects were examined clinically and parasitologically. Detection of hepatitis B seromarkers was also done. The infected subjects were treated with praziquantel at a dose of 60 mg/kg of body weight which was repeated after 4 weeks. Serum hyaluronic acid and procollagen III peptide were measured by radioimmunoassay. High hyaluronic acid was encountered in infected subjects when compared to their respective age-matched controls. Significant decrease of 4 and 8 weeks post-treatment was noted when compared to ist level before treatment. There was no significant change in serum procollagen III peptide on comparing infected subjects to their controls, whereas a significant increase was observed in its level after 4 and 8 weeks post-treatment compared to that before treatment. This work suggests that early treatment of intestinal schistosomiasis with specific chemotherapy (praziquantel) decreases serum hyaluronic acid and increases procollagen III peptide probably via downregulation of granulomatous inflammatory cell reaction and activation of collagenase enzymes, respectively.

PATTERN OF HBsAg POSITIVITY IN SELECTED GROUPS AT KING KHALID GENERAL HOSPITAL - HAIL REGION, KINGDOM OF SAUDI ARABIA.

BACKGROUND: Hepatitis B virus (HBV) is endemic in the Kingdom of Saudi Arabia. The acquisition of infection occurs early in life. The availability of safe and efficacious vaccines has led to the feasibility of an effective control of HBV infection. This study compares the pattern of HBsAg positivity among selected groups of patients with similar groups in other regions in the Kingdom of Saudi Arabia. OBJECTIVES: This study is conducted to determine the prevalence of HBsAg positive subjects among selected groups of patients attending the main general hospital in the Hail region. SUBJECTS AND METHODS: This retrospective study included 14029 subjects that were tested for HBsAg at King Khalid General Hospital, during the period from April 1994 to April 1996. RESULTS AND DISCUSSION: The overall prevalence of HBsAg positive subjects was 3.5%. The prevalence for Saudis was about 3.2% for Saudis. Pakistanis had the highest prevalence of 11.8%. The prevalence was 3.02% for blood donors, and 2.1 % among pregnant women. No significant difference was found among different age groups. Hail region had a lower prevalence of HBsAg positive cases compared to that estimated by previous studies in the Kingdom, suggesting success in efforts applied by the Ministry of Health (110H) for prevention of Hepatitis B viral infection. CONCLUSION: We can conclude that the prevalence of HBsAg positive subjects is declining in the Hail region, Kingdom of Saudi Arabia (KSA). This decline suggests the success of preventive efforts such as vaccination of newborns with the HBsAg vaccine. However, screening of pregnant women for HBsAg scents to be necessary.

Study of interleukin-2 receptor in schistosomiasis mansoni and its analogous changes in collagen diseases and schistosomal arthropathy.

An enzyme linked immunosorbent assay was used to quantify soluble interleukin-2 receptor (sIL-2R) in the serum of patients with different stages of S. mansoni infection, rheumatoid arthritis, systemic lupus erythematosus (SLE) and schistosomal arthropathy. The results demonstrated significant higher level of sIL-2R in different patient groups compared to the control group. The highest level of sIL-2R was recorded in hepatosplenic schistosomiasis complicated with ascites. The difference was statistically significant compared to other groups. There was no significant difference in sIL-2R regarding rheumatoid arthritis and SLE. Schistosomal arthropathy group showed significant higher level of sIL-2R compared to rheumatoid arthritis, SLE and early S. mansoni infection while the difference was insignificant compared to hepatosplenic schistosomiasis without ascites.

Evaluation of specific Fasciola antigen in the immunodiagnosis of human fascioliasis in Egypt.

Enzyme-linked immunosorbent assay (ELISA) and enzyme linked immunoelectrotransfer blot technique (EITB) were employed for the detection of circulating Fasciola antibodies in infected human sera using a specific Fasciola antigen, prepared by immunoaffinity purification of homogenates of Fasciola hepatica adult worms. Ninety two individuals diagnosed clinically and parasitologically were classified into: Fascioliasis group (21 patients), schistosomiasis group (21 patients) and subjects harbouring other parasitic infections (50 patients). Eighteen healthy individuals served as normal controls. ELISA was 100% sensitive and 93% specific with 96.5% diagnostic efficacy, whereas EITB was 100% sensitive and specific with 100% diagnostic efficacy. Our data revealed that ELISA can be used as a good screening test while EITB can serve as a confirmatory test for immunodiagnosis of fascioliasis.

Association between HLA-A, B, C and DR antigens and clinical manifestations of Schistosoma haematobium in the bladder.

This study is a search for the genetic susceptibility of Egyptians to Schistosoma haematobium infestation with its various bladder complications, including cancer. 80 bilharzial patients, 20 with simple bilharzial bladder cystitis, 30 with bilharzial bladder lesions, and 30 with bilharzial bladder cancer, as well as 35 normal Egyptian controls were studied. All patients were typed for HLA-A, B, C and DR antigens using the microlymphocytotoxicity test. HLA-A9 and its split Aw24 antigens were found to be negatively associated with the disease. As for the antigens with positive associations, HLA-B7 was significantly increased in the simple bilharzial cystitis group. In the bilharzial bladder cancer group, HLA-B16 and Cw2 antigens had positive associations. These findings might support the genetic control of the disease or the presence of an immune response and/or immune suppression genes which are in linkage disequilibrium with these HLA antigens and they control the susceptibility and pathological sequences of the disease.

Granulomatous cystitis: a reversible lesion in children.

27 children with granulomatous cystitis associated with schistosomiasis and urinary tract infection were studied by urine culture and the direct immunofluorescence technique to detect the presence of antibody-coated bacteria (ACB). All patients had urinary schistosomiasis (100%); urine cultures of more than 10,000 bacteria/ml were present in 18 (66%) patients, and ACB-positive tests in 23 (85%) patients. Depending upon an ACB-positive test, children received full courses of antimicrobial and antibilharzial therapy. Follow-up after 3 months showed that granulomatous lesions subsided in 20 out of 23 (87%) children.

Evaluation of the ELISA test as an epidemiological tool in schistosomiasis.

The ELISA test was studied to assess its applicability in epidemiological surveys of schistosomiasis. Specimens of serum, stool and urine from persons attending an outpatient clinic in an endemic area for S. mansoni and S. haematobium in Egypt were tested in the laboratories of the Theodor Bilharz Research Institute, Cairo. The ELISA test results were compared with parasitological findings and with the results of indirect immunofluorescence and indirect hemagglutination tests as reference data. The ELISA test showed an excellent sensitivity and specificity for both infections; the reading and recording of the extinction rates can be automated by using in time and money. The test can be strongly recommended for epidemiological surveys.