Clearance of HBV DNA in immunized children born to HBsAg-positive mothers, years after being diagnosed with occult HBV infection.

In a previous study, we observed immunoprophylaxis failure due to occult hepatitis B virus (HBV) infection (OBI) despite the presence of adequate levels of anti-HBs in 21 (28%) of 75 children born to HBsAg-positive mothers. The aim of the study was to explore the maintenance of this cryptic condition in this population. Of 21 OBI-positive children, 17 were enrolled. HBV serological profiles were determined by enzyme-linked immunosorbent assay. Highly sensitive real-time and standard PCR followed by direct sequencing were applied in positive cases. The mean age (±SD) of studied patients was 6.57 ± 2.75 years. All children still were negative for HBsAg. All but one (94%) were negative for HBV DNA. Only two children were positive for anti-HBc. The results of the most recent anti-HBs titration showed that 4 (23.5%) and 13 (76.5%) had low (<10 IU/mL) and adequate (>10 IU/mL) levels of anti-HBs, respectively. The only still OBI-positive patient had an HBV DNA level of 50 copy/mL, carried the G145R mutation when tested in 2009 and again in 2013 in the 'a' determinant region of the surface protein. Further follow-up showed that after 18 months, he was negative for HBV DNA. In high-risk children, the initial HBV DNA positivity early in the life (vertical infection) does not necessarily indicate a prolonged persistence of HBV DNA (occult infection). Adequate levels of anti-HBs after vaccine and hepatitis B immune globulin immunoprophylaxis following birth could eventually clear the virus as time goes by. Periodic monitoring of these children at certain time intervals is highly recommended.

Outcome of adult onset systemic lupus erythematosus in Iran.

AIM: The aim of this study was to determine systemic lupus erythematosus (SLE) survival in adult patients and its predictors in Iran. METHODS: The adult patients diagnosed with SLE and admitted to our referral general hospital from 1992 to 2011 were studied. Demographic, clinical and laboratory data at the time of diagnosis were obtained retrospectively and analyzed. Survival rates were calculated by the Kaplan-Meier method. Predictors of mortality were assessed by Cox regression analysis. RESULTS: In total, 417 were enrolled in the study; 23 were lost to follow-up. Mean (SD) age of SLE onset was 30 (9.7) years. During the study period 35 patients (8.9%) died. The most common causes of death were active SLE (43%), infections (28.6%) and circulatory diseases (20%). Overall survival rates after 5, 10, 15 and 20 years were 93%, 90%, 90% and 80%, respectively. Poor survival predictors in univariate analysis were pericarditis, seizure and hematuria. With multivariate Cox regression analysis, no pericarditis (p = 0.007, HR = 0.22, 95%CI: 0.075-0.657) and no seizure (p = 0.019, HR = 0.35, 95%CI: 0.149-0.846) at the time of SLE diagnosis were found as protective factors in patients' survival. CONCLUSION: Our study revealed that the survival rate of SLE is comparable with the acceptable worldwide trend. Presenting with pericarditis and seizure at the time of SLE diagnosis prominently decreased the survival rate. Prospective and multicenter studies are needed to better identify the behavior of SLE in Iran.

Hepatitis B virus surface protein mutations clustered mainly in CTL immune epitopes in chronic carriers: results of an Iranian nationwide study.

Mutations within the coding region of hepatitis B surface antigen (HBsAg) have been found naturally in chronic carriers. To characterize the mutations of HBsAg from Iranian chronic carriers who were vaccine and/or medication naive. The surface genes from 360 patients were amplified and directly sequenced. The distribution of amino acid substitutions was classified according to different immune epitopes of the surface protein. All isolates belonged to genotype D. 222 (61.6%) of 360 patients contained at least one amino acid substitution. 404 (74.5%) of 542 amino acid changes occurred in different immune epitopes of HBsAg, of which 112 (27.7%) in 32 residues of B-cell epitopes (62 in the 'a' determinant); 111 (27.4%) in 32 residues of T helper; and 197 (48.7%) in 32 residues inside cytotoxic T lymphocyte (CTL) epitopes. One Th (186-197) and two CTL (28-51 and 206-215) epitopes were found to be hotspot motifs for the occurrence of 213 (52.7%) substitutions. 20 stop codons were identified in different epitopes. There was a significant association between amino acid substitutions and anti-HBe seropositivity; however, the correlation between such changes with viral load and ALT levels was not significant. In chronic hepatitis B virus(HBV) carriers, positive selection in particular outside the 'a' determinant appeared to exert influence on the surface proteins. These changes could be immune escape mutations naturally occurring due to the host immune surveillance especially at the T-cell level.

Spatio-temporal variations of extract produced and fatty acid compounds identified of Gundelia tournefortii L. seeds in central Zagros, Iran.

This study was performed to fulfill two aims. The first aim was to isolate the seed extract of Gundelia tournefortii L. at two phenological stages of seed production (the beginning and end of seed production); the second one was to identify the fatty acid compounds of G. tournefortii L. seeds in its major habitats located in the Central Zagros region, Iran. Among them, some of the major environmental factors on the reproductive growth stage i.e., physiography, soil and climate were studied. Extraction was performed using the Soxhlet apparatus, and the fatty acid compounds were identified by The GC-FID analysis. As a result, site No. 5 with the values of 6.06 and 7.21 g had the highest amount of extract produced, while sites number 7 and 8 had the least one which was 2.86 and 3.84 g at two phenological stages of seed production. There was a strong correlation among the major environmental variables and the amount of extract produced in the phenological stages of seed production; this was also confirmed in relation to the fatty acid compounds and some of their characteristics. Overall, the efficacy of environmental factors on the synthesis process of secondary metabolites is undeniable.

Validation of an in-vitro method for Hepatitis B vaccine potency assay: specification setting.

AIM: Production batches of the Hepatitis B vaccine should be tested by the National Control Laboratory (NCL) before being released to the market, in terms of their potency. This can be done either by means of the mouse immunogenicity (in-vivo) method, which is a time-consuming and labor intensive process, or by an in-vitro method with acceptable analytical performance and with specifications determined based on the results obtained from testing some production batches of the vaccine with proven efficacy. Here we report the feasibility of using and validation of a commercial enzyme-linked immunosorbent assay (ELISA) kit replacing the manufacturer's method and setting of different specification for potency of the particular vaccine. METHODS: For the in-vitro potency assay of the Hepavax-Gene®, produced by Berna Biotech Korea Corp, a commercial ELISA kit for hepatitis B surface antigen (HBsAg) quantitation (Hepanostika® HBsAg Ultra from Biomerieux) was used to determine the relative potency. Validation parameters were evaluated following the International Conference on Harmonization (ICH) guidelines. Specification of the vaccine potency was determined based on the results generated by the commercial ELISA kit. Some batches were tested by in-vivo method as well. RESULTS: It was confirmed that the ELISA kit, when used for vaccine potency testing, meets the criteria for accuracy (80% to 110% recovery), precision (repeatability, with a CV% less than 5%; and intermediate precision, with a CV% less than 10%) and Linearity (r2> 98%), as well as being able to detect HBsAg specifically. Specification of the in-vitro method was also determined as having a relative potency of >50%. CONCLUSION: The Hepanostika® HBsAg Ultra kit from Biomerieux can be used to determine the relative potency of the Hepavax-Gene® Hep B vaccine as an alternative to the manufacturer's method and with different specifications.