negin nikouyan

In spite of recent progress in mRNA technologies and their potential applications for treatment of human diseases, problems such as the transient nature of mRNA limit the stability of gene up-regulation and, thus, potentially reduce mRNA efficiency for gene therapy. Using human β-globin 5′ and 3′ untranslated regions (UTRs), this study aimed to develop the different chimeric constructs of mRNAs to increase the stability of destabilized green fluorescent protein (EGFPd2) in HEK 293 cells. Purified human β-globin (HBG) 5′-3′UTRs, and the coding sequence of destabilized green fluorescent protein (EGFPd2) were amplified separately and ligated to each other using SOEing PCR method in a different format. As controls, the original construct of EGFPd2 under the control of T7 promoter was used. Following in vitro transcription, HEK 293 cells were then transfected with several constructs and incubated at 37oC in a CO2 incubator. They were monitored under a fluorescence microscope every four hours for the first 24 hr, then every 12 hr afterwards. The resulting fluorescence was measured as a surrogate for translation efficiency and duration. By monitoring the HEK cells over 48 hr, cells transfected with mRNA with various HBG UTRs showed significantly different fluorescence intensity and stability in comparison with the pEGFPd2 prototype (control transcript) overtime. Overall, the images show that replacement of the 3′ UTR end of the prototype vector pGFPd2 with the 3′ end of β- globin mRNA increases the half-life of the chimeric mRNA for more than 32 hr. This result indicates that β-globin 3′ UTR would definitely increase the half-life of mRNA and may help to decrease the mRNA therapeutic dosage in the treatment of diseases associated with mRNA therapy.

There are few studies indicating the post-neonatal HBV vaccination level of anti-HBs antibody in first-year enrolled university students in Iran. In addition, anti- HBc antibody detection, which is a good indicator of virus exposure, has not been reported in vaccines. Hence, this study was conducted to determine the level of anti-HBs and anti-HBc antibodies in the serum sample of medical laboratory students who had received primary infantile HBV vaccination.
This study was conducted on first-year students enrolled in the department of laboratory sciences at Shiraz University of Medical Sciences, Iran. For determining anti-HBs and anti-HBc titers, 5 mL of venous blood was aseptically collected. Anti-HBs and anti-HBc antibody levels were determined by enzyme-linked immunosorbent assay. HBV DNA was also performed on DNA extracted from individuals positive for an anti-HBc antibody test.
Of the 257 vaccinated individuals (188 females and 69 males) who participated in this study, 36.2% showed a non-protective anti-HBs response (anti-HBs Our results indicate that a substantial number of our study population vaccinated against HBV during childhood showed non-protective anti-HBs antibody level. Therefore, a booster dose of vaccine needs to be scheduled for students with anti-HBs level

Epidemiological studies have reported commonly distributed transfusion-transmitted virus (TTV) in different populations with parental risk factors, including human immunodeficiency virus (HIV).
This study was performed to determine and compare the prevalence of TTV infection among HIV-positive patients and healthy blood donors.
Patients and A total of 186 HIV patients and 165 healthy blood donors with no markers of HIV infection were included in this study during 2004 - 2012. Semi-nested Polymerase Chain Reaction (PCR) assay was performed for the detection of TTV DNA. HBV surface antigen (HBsAg), Hepatitis C Virus Antibody (HCVAb), and CD4 were investigated in the sera of HIV-positive subjects. Aspartate transaminase (AST) and Alanine transaminase (ALT) levels were also measured.
The mean age of HIV-positive and healthy subjects was 39.3 and 40.5 years, respectively. In total, 182 (97.9%) subjects were male and 4 (2.1%) were female. TTV DNA was detected in 35 out of 186 HIV patients (18.8%; 95% CI, 13.2 - 24.4%). The prevalence of TTV in the HIV group was significantly higher (P = 0.027) than blood donors (11%; 95% CI, 6.2 - 15.8%). Age, marital status, unsafe sexual activity, and use of injection drugs were not significantly associated with the prevalence of TTV infection in HIV patients.
Considering the higher frequency of TTV infection in HIV patients in comparison to healthy blood donors, HIV infection may be an important risk factor for TTV infection. In addition, the lower frequency of TTV infection in healthy individuals in comparison to HIV patients reveals the transmission of TTV infection via routes other than blood and drug injection; therefore, the effect of fecal-oral route needs to be examined in future studies.

Infection with hepatitis E virus (HEV) is endemic in developing countries and reveals significant regional differences. Several studies have reported virus transmission via blood transfusion. To date, however, no cases of HEV RNA detection in blood donors have been reported from Iran.
The aim of this study was to determine the presence of HEV RNA in plasma samples of blood donors referred to a blood transfusion center in Shiraz in the southwest of Iran. The HEV genotypes were also investigated using nucleotide sequencing.
Patients and Blood samples were collected from 700 blood donors who were referred to Fars blood transfusion organization from January to March 2014. Plasma samples were screened for the presence of HEV IgG and IgM antibodies by standard enzyme immunoassay. Samples seroreactive to anti-HEV were further tested for the presence of HEV RNA using nested polymerase chain reaction (PCR) with universal primers for detection of all four HEV genotypes. Positive PCR samples were then subjected to DNA sequencing for further analysis.
Fifty (50, 7.1%) out of 700 plasma samples tested positive for anti-HEV antibodies. HEV RNA was detected in 7/50 (12%) of the antibody-positive samples, the majority of which were IgM positive. Sequence analysis of seven isolates of the HEV RNA ORF 2 gene region revealed > 80% similarity with genotype 1.
The analysis indicates that the HEV isolated from blood donors in the southwest of Iran belongs to genotype 1. However, more samples from other geographic regions of Iran are needed to confirm these findings. Because transmission of HEV by administration of blood or blood components is likely to occur, it may be sensible to screen donor blood for HEV to eliminate transfusion-transmitted HEV infection when the recipient is immunocompromised.

Viral load measurements are commonly used to monitor HCV infection in patients with chronic diseases or determining the number of HCV-genomes in serum samples of patients after sustained virological response. However, in some patients, HCV viral load in serum samples is too low to be detected by PCR, especially after treatment.. The aim of this study was to develop a highly specific, sensitive, and reproducible in-house quantitative PCR using specific primers and probe cited in highly conservative region of HCV genome that allows simultaneous detection of HCV genotypes 1 - 4.. In this study, three sets of primer pairs and a TaqMan probe for amplification and detection of selected region within 5’-non-coding (5’NCR) of four HCV genotypes were used. Using plasmid containing 5’NCR region of HCV, standard curve, threshold, and threshold cycle (CT) values were determined. Real-time and nested PCR were performed on HCV genotypes 1 - 4 extracted from plasma and peripheral blood mononuclear cells (PBMCs) samples collected from patients with chronic HCV infection.. The lower limit detection of this in-house HCV real-time RT-PCR was determined as 100 RNA copies/mL. Inter- and intra-assay coefficient of variation (CV) of this in-house HCV real-time RT-PCR ranged from 0.9% to 1.8% and 1.76% to 3.94%, respectively. The viral load of the genotyped samples ranged from 2.0 × 106 ± 0.31 to 2.7 × 105 ± 0.46 copies/mL in serum samples and 5 × 102 ± 0.36 to 4.0 × 103 ± 0.51 copies/106 cells/mL of PBMCs.. The quite sensitive in-house TaqMan real time RT-PCR assay was able to detect and quantify all four main HCV genotypes prevailing around all geographical regions of Iran..