Quercetin potentiates transdifferentiation of bone marrow mesenchymal stem cells into the beta cells in vitro.

PURPOSE: Type 1 diabetes is an autoimmune disease caused by the destruction of ß-cells in the pancreas. Bone marrow mesenchymal stem cells are multipotent and easy accessible adult stem cells that may provide options in the treatment of type 1 diabetes. Injured pancreatic extract can promote the differentiation of rat bone marrow mesenchymal stem cells into ß-cells. We aimed to observe the effect of quercetin in differentiation and insulin secretion in ß-cells. METHODS: Bone marrow mesenchymal stem cells were obtained from the tibiae of rats. Cell surface markers were analyzed by flow cytometry. The cells were treated with rat injured pancreatic extract and quercetin for 2 weeks. Insulin secretion was measured by ELISA. Insulin expression and some islet factors were evaluated by RT-PCR. PDX1, a marker for ß-cell function and differentiation, was evaluated by both immunocytochemistry and Western blot. ß-cell count was determined by stereology and cell count assay. RESULTS: ELISA showed significant differences in insulin secretion in the cells treated with RIPE + 20 µM quercetin (0.55 ± 0.01 µg/L) compared with the cells treated with RIPE alone (0.48 ± 0.01 µg/L) (P = 0.026). RT-PCR results confirmed insulin expression in both groups. PDX1 protein was detected in both groups by Western blot and immunocytochemistry. Stereology results showed a significant increase in ß-cell number in the RIPE + quercetin-treated cells (47 ± 2.0) when compared with RIPE treatment alone (44 ± 2.5) (P = 0.015). CONCLUSIONS: Quercetin has a strengthening effect on the differentiation of rat bone marrow mesenchymal stem cells into ß-cells and increases insulin secretion from the differentiated ß-cells in vitro.

Detection of Hepatitis C virus RNA in peripheral blood mononuclear cells of patients with abnormal alanine transaminase in Ahvaz.

PURPOSE: Hepatitis C virus (HCV) is an important agent for chronic and acute hepatitis. Occult hepatitis C remains a major health problem worldwide. Patients with chronic occult HCV may progress to cirrhosis and hepatocellular carcinoma. The aim of this study was to determine prevalence of occult hepatitis C by IS-PCR-ISH (in situ PCR in situ hybridisation) in the patients with abnormal ALT. MATERIALS AND METHODS: The blood samples were taken from 53 patients including 17 females (32.1%) and 36 (67.9%) males who had abnormal alanine transaminase (ALT) for more than 1 year. The mean ALT and aspartate transaminase (AST) level were 41.02±9.3 and 24.17±7.3, respectively. The patients' age were between 4 and 70-years old with mean age 38±13. All the patients were negative for HCV antibody, HCV RNA and HBs Ag. The peripheral blood mononuclear cells (PBMC) were separated with ficoll gradient from each blood sample, then the cells were fixed on slides by cold acetone and followed by IS-PCR-ISH for HCV RNA detection. RESULTS: Seventeen (32%) patients including 6 (11.3%) females and 11 (20.7%) males showed positive results for HCV RNA by in situ-PCR in situ hybridisation. Ten (18.8%) positive cases were between 20 and 40-years old and 6 (11.3%) positive patients were between 40 and 60 years old. Ten (19.6%) patients who were positive for IS-PCR-ISH also had positive anti-HBc IgG and 7 (13.2%) patients were negative for HBc-IgG. CONCLUSION: In the present study high rate of 32% occult hepatitis C were found among the patients with elevated ALT.

Expression and Purification of Recombinant Mycobacterium Tuberculosis (TB) Antigens, ESAT-6, CFP-10 and ESAT- 6/CFP-10 and Their Diagnosis Potential for Detection of TB Patients.

BACKGROUND: One of the most widely used methods to detect tuberculosis (TB) infection is the tuberculin skin test (TST). The completion of Mycobacterium tuberculosis (M. tuberculosis) genome sequence has led to identification of several antigens that can be utilized for accurate diagnosis and control of TB. The aim of this study was to purify the recombinant M. tuberculosis antigens for the evaluation of their potential in TB diagnosis. METHODS: The recombinant secretory antigens, ESAT-6, CFP-10 and ESAT-6/CFP-10 were produced by PCR and cloning methods. To investigate antigen specific responses of these recombinant antigens in detection of TB, ex vivo enzyme linked immunospot (ELISPOT) test in 30 clinically diagnosed TB patients was evaluated. RESULTS: The selected M. tuberculosis antigens were cloned, expressed and purified in Escherichia coli (BL21). ELISPOT assay for detection of TB showed the sensitivity of 93, 90 and 100% for recombinant ESAT-6, CFP-10 and ESAT-6/CFP-10 proteins respectively, which is significantly higher than conventional TST. CONCLUSION: The recombinant antigens of ESAT-6, CFP-10 and ESAT-6/CFP-10 can be used as an accurate means of detecting TB in Iran.

C-terminal-binding protein interacting protein binds directly to adenovirus early region 1A through its N-terminal region and conserved region 3.

C-terminal-binding protein interacting protein (CtIP) was first isolated as a binding partner of C-terminal-binding protein (CtBP). It is considered to contribute to the transcriptional repression and cell cycle regulatory properties of the retinoblastoma (Rb) family of proteins and to have a role in the cellular response to DNA damage. Here, we have shown that CtIP is a novel target for the adenovirus oncoprotein early region 1A (AdE1A). AdE1A associates with CtIP in both Ad5E1-transformed cells and Ad5-infected cells and binds directly in glutathione-S-transferase pull-down assays. Two binding sites have been mapped on Ad5E1A - the N-terminal alpha-helical region (residues 1-30) and conserved region 3 (CR3) - the transcriptional activation domain. CtIP can bind AdE1A and CtBP independently, raising the possibility that ternary complexes exist in Ad-transformed and -infected cells. Significantly, reduction of CtIP expression with small interfering RNAs results in reduction of the ability of a Gal4 DNA-binding domain-CR3 construct to transactivate a Gal 4-responsive luciferase reporter and this effect is reversed by reduction of CtBP expression. Therefore, in this model, CtIP acts as a transcriptional co-activator of AdE1A when dissociated from CtBP, through the action of AdE1A. These data are consistent with observations that CtIP expression is induced by AdE1A during viral infection and that reduction of CtIP expression with RNA interference can retard virus replication. In addition, AdE1A causes disruption of the CtIP/Rb complex during viral infection by its interaction with CtIP, possibly contributing to transcriptional derepression.

Normal serum prostate specific antigen levels in men in Yasuj province, Islamic Republic of Iran.

We assessed serum prostate specific antigen (PSA) levels in 650 men over 40 years referred to 3 Yasuj hospitals for blood cell count in 2003/2004. Men affected by prostate cancer, prostatitis or transurethral instrumentation were excluded. PSA was determined by an immunoassay technique. PSA levels in different age groups were: 40-49-year-olds--mean = 0.7 ng/dL, normal = 0-1.35 ng/dL; 50-59-year-olds--mean = 0.9 ng/dL, normal = 0-1.85 ng/dL; 60-69-year-olds--mean = 1.6 ng/dL, normal = 0-3.2 ng/dL; > or = 70-years-olds--mean = 2.3 ng/dL, normal = 0-4.4 ng/dL. Normal PSA levels in our society were lower than those in the United States, Europe and Japan.

Normal serum prostate specific antigen levels in men in Yasuj province, Islamic Republic of Iran

We assessed serum prostate specific antigen [PSA] levels in 650 men over 40 years referred to 3 Yasuj hospitals for blood cell count in 2003/2004. Men affected by prostate cancer, prostatitis or transurethral instrumentation were excluded. PSA was determined by an immunoassay technique. PSA levels in different age groups were: 40-49-year-olds-mean = 0.7 ng/dL, normal = 0-1.35 ng/dL; 50-59-year-olds-mean = 0.9 ng/dL, normal = 0-1.85 ng/dL; 60-69-year-olds-mean = 1.6 ng/dL, normal = 0-3.2 ng/dL; >/= 70-years-olds-mean = 2.3 ng/dL, normal = 0-4.4 ng/dL. Normal PSA levels in our society were lower than those in the United States, Europe and Japan

Analysis of arylsulfatases A and B, acid phosphatase, lactate dehydrogenase, and aspartate transaminase in chronic periapical lesions of endodontic origin.

Attempts were made to detect and measure the activities of arylsulfatases. A&B acid phosphatase, lactate dehydrogenase, and glutamate oxaloacetate transaminase (aspartate transaminase) enzymes in human chronic lesions of endodontic origin. Thirteen periapical lesions of endodontic origin and 11 noninflamed control periapical tissues were obtained. The specimens were carried to the laboratory on liquid nitrogen and kept at -70 degrees C. Samples were thawed, homogenized, and then assayed for enzyme activities. The specific activities of arylsulfatase A (nmol/hr/mg protein) were 55.0+/-10.7 (chronic lesions) vs. 3.4+/-2.2 (controls) (p < 0.01). Arylsulfatase B specific activities (nmol/hr/mg protein) were 50.3+/-6.4 (chronic lesions) vs 91.8+/-18.4 (controls). Total acid phosphatase activities (mU/mg protein) were 45.8+/-6.6 (chronic lesions) vs. 26.8+/-3.1 (controls). Lactate dehydrogenase activities (Berger-Broida units/mg protein) of the chronic periapical lesions were significantly higher than the control group (362+/-63.2) vs. (140+/-46.0) (p < 0.05). There was no significant difference between the specific activities of aspartate transaminase in chronic lesions and the control group (68.0+/-14.5) vs. (53.0+/-10.4) mU/mg protein).

Modulation of the pituitary and basomedial hypothalamic lysyl-aminopeptidase activities be beta-estradiol and/or an aqueous extract of Physalis alkekengi fruits.

Injections of an aqueous extract of winter cherry fruits (Physalis alkekengi) to adult female cycling rats by an intraperitoneal route resulted in the diminution of the pituitary lysyl-aminopeptidase (Lys-AP) activity by 50% and that of the basomedial hypothalamus (BMH) by 45%. Administration of daily doses of 3.75, 7.5, and 15 micrograms beta-estradiol for a period of 5-8 days to such animals increased pituitary Lys-AP activity from 31% to 61.5% and that of BMH from 20% to 87%, respectively. Administration of the same doses of beta-estradiol along with a given dose of the aqueous extract for 7-8 days diminished Lys-AP inhibitory effect of the extract in both the pituitary and BMH and eventually, at the highest dose of beta-estradiol, increased the pituitary enzyme activity by 9% and that of BMH by 5%. It is concluded that Lys-AP enzymes of both tissues, being estrogen-induced proteins, are inhibited by the estrogen antagonistic principle of the winter cherry aqueous extract. It is further suggested that BMH Lys-AP activity may be used as an enzyme marker for the action of beta-estradiol in hypothalamus.