Abstract

 Background and Objectives

 The most important goal of IBTO is to prepare safe and sufficient blood and blood components thus, the appropriate screening of donors out of low-risk population is significant. It is likely that women population compared with men is at lower risk in regard to high-risk behaviors leading to blood-transmitted infections. However, the donation attempts on part of women compared to men are less frequent.

 Materials and Methods

  A cross-sectional study was conducted on Iranian female population at the age range of 17-65 in eight provinces of Iran. A questionnaire was prepared. The number of samples was calculated as 12000 using statistical formulas. The sampling method was multi-stage cluster. Finally, the data were analyzed using SPSS 11 statistical software.

 Results

 The age average of women under study was 32.6 ± 12.1. Most of them were married, housekeeper, and had diploma. 24.1% of them had a record of blood donation while 75.4% never enjoyed such an experience. The educational background and employment rate of women with no blood donation precedent were significantly lower than those with previous history of blood donation (P<0.001). The most frequent reason for women^,s unwillingness to embark no blood donation was considered to be their fear of being infected with infectious and blood-borne diseases. On the whole, 75.2% and 24.8% of women under study showed respectively a negative and positive attitude toward blood donation.

 Conclusions

 Since fear of being infected with infectious and blood-borne diseases out of blood donation is somehow the outcome of the lack of awareness of the public about transfusion medicine thus, the most significant reason for the lack of donation can be attributed to the lack of knowledge on the part of women regarding transfusion medicine. Based on the findings of the present study, it is recommended that training about the significance of blood donation and women^,s acceptance criteria for blood donation be promoted extensively through TV and Radio broadcasting.

 Key words: Blood donation, Blood Transfusion Organization, Women, Attitude

  Abstract

 Background and Objectives

 Post -transfusion hepatitis B may occure if donors donate blood in the window period, in the early convalescence phase and period with very low levels of HBsAg in blood.

 Case

 In December 2003, a case as post- transfusion hepatitis B in a blood recipient was reported to IBTO.The patient had received 2 units of red blood cells.Trace back program was set up to find out the donors possible involvement in viral transmission.

 Conclusions

 Donors and recipients were not positive for HBsAg and Anti-HBc IgM so current or recent infection during last 6 months had to be excluded. This case report explains a successful trace back , and accurate well- maintained records.

  Key words: Trace back, Hepatitis B, Transfusion

  Abstract

 Background and Objectives

  Screening the blood donors for serological markers reduced the incidence of transfusion-transmitted infections especially post-transfusion hepatitis C. However, there remains residual risk due to pre-seroconversion period. HCV RNA (PCR) of blood donations reduced the residual risk of transfusion-transmitted HCV infection. In this study, blood donations were screened for HCV RNA by RT-PCR method.

  Materials and Methods 

  An extra plasma sample was collected from 1026 blood donors. 1000 out of 1026 samples were negative for HBsAg, anti-HCV (EIA, third generation), anti-HIV and RPR. Every 5 samples were pooled. The sensitivity of HCV-RNA detection by RT-PCR method was 380 geq/ml according to Proficiency VQC panel. 1000 donations in 200 pools were tested.

 Results

 False reactivity of samples considered positive accounts for 5.5% of cases, and 5.5% were invalid due to non-specilic bands. 6% of the pools were false-positive. A false positive result was defined as positive on initial testing but negative on repeat single testing. However, all of the samples were negative for HCV RNA by RT-PCR method.

 Conclusions

 No sample was found to be serologically negative and HCV RNA positive. However, further studies are recommended for further clarification.

  Abstract

 Background and Objectives

  In this study our aim was to determine HLA-Class I and II antigens freguencies of Hamedani ethnic group. In addition to demographic studies and disease association, it has wide application in bone marrow donor registeries.

  Materials and Methods 

  In order to establish DNA-based HLA typing in central Laboratory of Iranian Blood Transfusion Organization, a comparison of serological and molecular (sequence specific primers “SSP”) methods for HLA-DRB has been performed. The study was descriptive and the population under study were selected out of the native people of Hamedan 100 healthy volunteer blood donors were chosen by questionnaire. 10ml heparinized and 3ml EDTA blood were collected from each selected donor. EDTA (PCR) samples were then frozen.

 Results

  N.I.H standard microlymphocytotoxicity and Nylon wool T and B cells separation was used for serological I and II typing. HLA-Class I plates were prepared from Iranian Blood Fractionation and Research Company and for Class II we used Biotest DR/DQ Typing Trays. PCR was done using “Roche high pure DNA extraction” Kit and HLA-DRBSSP (Biotest). The most and least frequent HLA-B antigens were B5 group (B51/B52) and B16 (38,39) respectively. Because of low resolution of HLA-DRB Kit, no significant difference was observed between serological and PCR methods. Although some blanks have been determined by PCR.

 Conclusions

  The HLA-DRB determination by PCR is mandatory for donor/recipient pairs (even sibling) for bone marrow transplantation for donors it should be done by high resolution kits.

Abstract

Background and Objectives

After hepatitis and AIDS, malaria is the most prevalent transfusion outcome in endemic areas. Presence of asymptomatic carriers of malaria parasites in the endemic areas can be a source of infection in transmission of malaria by blood transfusion. Prevention of malaria caused by blood transfusion depends on screening blood donors and deleting infected blood samples. To screen blood samples, parasitological, serologic and molecular methods have been applied.

Materials and Methods

In this study 120 blood donors in Iranshahr in Sistan-Baloochestan province were tested with different methods of thick and thin blood films, Immuno-Fluorescent Antibody Test (IFAT), and Polymerase Chain Reaction (PCR).

Results

The result of all thick and thin blood films were negative. IFAT by using P.vivax antigen and P.falciparum antigen for 38 and 6 donors respectively showed a titre of antibody equal to
± 1/20-1/320 (17 of the former group and 4 of the latter had a history of malaria infection). The PCR assay using silica for DNA extraction and using P.falciparum specified primers with sensitivity rate equal to 2-3 parasites per microlitre of blood was negative for all subjects under study.

Conclusions

This study showed, although microscopic examination of blood smears was inexpensive and simple, but it is labor-intensive and time-consuming that makes it insensitive for detection of low-level parasitemia in asymptomatic donors and for screening a large number of specimen. IFAT would not always show the real existence of parasites and in spite of simplicity and sensitivity because of its disability to be automated is not suitable for screening a large number of specimen. On the other hand, IFAT in individuals with malaria history and absence of parasites in their blood may be positive for a long period. It was approved that molecular methods such as PCR were more sensitive and more specific than conventional microscopic examination and their great advantage was the ability to detect the infection with low-level parasitemia that may have been distinguished by blood films examination. In the present study, probably because of low number of specimen or limited study duration with PCR method, or probably since parasitemia exiting in the subjects under study was less than 2-3 parasites per microlitre of blood, we were not able to detect positive cases.

Key words:  Malaria, Blood transfusion, PCR, IFA, Blood donors

  Abstract

 Background and Objectives

  The most important challenge in selecting suitable assays for the detection of anti-HCV is sensitivity. In this study, 20 assays (EIA method) were compared with each other and with anti-HCV 3.0 Enhanced SAV (Ortho Company production) as the reference assay recommended by WHO .

  Materials and Methods

  20 kits were compared by 3 to 4 seroconversion and 2 to 3 performance panels. The relative sensitivity of kits was calculated based on WHO recommendations.

 Results

  In seroconversion panels, relative sensitivity of 3 assays was the same as the reference assay and 5 assays showed lower relative sensitivity, but the differences between these five kits and the reference assay appeared just in two samples. In performance panels, two assays came out to be the same as the reference assay and the other 5 assays detected just 2 samples to have a level lower than anti-HCV 3. In all seroconversion and performance panels, the best results were obtained by ETI-AB-HCH-K4 (146) (Diasorin), Monalisa Anti-HCV plus Version 2 (BIO-RAD), Hepanostica Anti-HCV ULTRA (BIOMERIEUX), Anti-HCV-EIA 3^rd (Avicenna Medial Center), and HCV AB (DIA PRO).

 Conclusions

  For improvement of blood safety, the assay with high sensitivity is recommended to be used, and the samples with weak positive reactions especially in seroconversion and low titer performance panels should be given more attention.

 Key words: Antibody , Hepatitis C, Elisa, Sensitivity, Seroconversion panel

  Abstract

 Background and Objectives

  The prevalence of GBV-C and HGV in blood donor populations in developd countries based on HGV RNA detection and anti-E₂ screening ranges from 1 to 5 and 3 to 14% respectively. The aim of this study was to investigate seroepidemiologic hepatitis G virus (HGV) in blood donors, haemodialysis patients, haemophiliacs, and β thalassemics with a history of liver disease by Elisa tecnique.

  Materials and Methods

  In this descriptive study, blood samples of 330 volunteer blood donors, 44 haemodialysis patients, 16 haemophiliacs, and 40 β major thalassemics with a history of liver disease were studied by Elisa technique for their seroepidemiologic status of hepatitis G virus and their past record of HGV infection. For data analysis, Chi-square, Fisher exact test, and SPSS version 11.5 were used.

 Results

  This study showed that out of 330 healthy blood donors 14(4.2%), out of 44 haemodialysis patients 10(22.7%), out of 16 haemophiliacs 5 (30.3%) and out of 40 β thalassemics 10 (25%) were positive for HGV-anti-E2. These data are significant evidence for HGV to be considered as a transfusion-transmitted infection. The prevalence of anti-HGV and anti-HCV (co-infection) was found to involve 10 (30.3%) of haemodialysis patients, 4 (28.6%) of haemophiliacs and 9 (23.7%) of β thalassemics. It was also found that 1 (8.3%) of haemodialysis patients, 1 (33.3%) of haemophiliacs, and 1 (50%) of β thalassemics were infected with anti-HGV and HBsAg co-infection.

 Conclusions

  The prevalence of HGV was high in multitransfused individuals including haemodialysis patients, haemophiliacs, and thalassaemics. Therefore, HGV was a transfusion-transmittable agent. Co-infection of anti-HGV with HCV was observed in viruses. It is recommended that further studies focus on evaluating sexual and vertical transmission routes so as to cast light on relatively high rate of HGV in donor population.

 Key words: HGV, Blood donors, Haemodialysis, Haemophilia, Beta thalassemia

  Abstract

 Background and Objectives

 The capsid or core Ag of Hepatitis C virus is a multifunctional protein which has the principal pathogenesis and diagnostic role in HCV related infections and most of these properties are attributed to the hydrophilic section (amino acids 2-122) of this protein. For different research and diagnostic applications, high amounts of this protein in pure and original form are required. So, the aim of this study was to clone the gene, optimize the expression condition, purify it in the original form, and immunologically characterize hydrophilic section of HCV Core Ag, expressed by T7-araBAD promoter system in E.coli.

 Materials and Methods

 The PCR amplified region corresponding to 2-122 section of this Ag from genotype Ib was cloned in pIVEX 2.3, a T7 promoter derived vector. The proper construct after digestional analysis and sequencing confirmations was transformed into BL21-AI E.coli, and protein expression under control of araBAD promoter by addition of 0.2% Arabinose was induced.

 Results

 After optimization of expression condition, purification of protein by NI-NTA agarose gel chromatography in native condition by immidazole yielded about 3.5mg/L of HCV core Ag. Immunological studies by western blotting through application of core specific mAbs and results of ELISA tests indicated that the protein is with desired immunological properties.

 Conclusions

 AraBAD promoter can be perfectly utilized to produce the hydrophilic section of HCV core in high yields, and purification through NI-NTA in native condition may provide the antigen for different research and diagnostic applications.

 Key words : Hepatitis C core Antigen, Arabinose, NI-NTA, Promoter 

  Abstract

 Background and Objectives

  Beta-2 microglobulin ( β 2MG) is the light chain of Histocompatibility–Class I human antigen and its normal range is <3mg/ml. β 2MG level in sera of hepatitis B patients increases. In Hepatitis infection the presentation of the viral antigen on the hepatocyte in the presence of Class I HLA antigen plays a major role in the elimination of the virus.

  Materials and Methods

  In this descriptive study, s β 2MG, HBsAg (by ELISA), and HBV DNA (by PCR) were evaluated in sera of 49 patients with hepatitis B and 35 subjects in control group .

 Results

  Our results showed HbsAg was positive in all patients. 29 of patients were HBV-DNA-PCR positive and 20 HBV-DNA-PCR negative. β 2MG in all subjects in control group was in normal range and in 34.7% of patients above normal limit. β 2MG in HBV-DNA-PCR positive patients was higher than HBV DNA PCR negative patients. Such differences were significant (p < 0.05) .

 Conclusions

  It seems S β 2MG is a good marker for HBV replication and its absence may exclude HBV replication. The role of β 2MG in monitoring response to therapy needs to be further evaluated.

 Key words: Hepatitis B virus, HBsAg , β 2MG (beta - 2 microglobulin), PCR , ELISA

  Abstract

 Background and Objectives

 Blood transfusion has a critical role in clinical practice, but there has always been a possibility for transmission of infections. A lot of research have been conducted recently to find the cause of these infections. In 1997, TTV was first found by Nishizawa. Our aim was to estimate the prevalence rate of TTV in healthy blood donors and recipients of blood components in Tehran and prepare PCR kits to detect this virus.

 Materials and Methods

  In this research, we studied the prevalence rate of TTV in 250 thalassemic, hemophilic and dialysis patients and 250 blood donors. After extraction of DNA and amplification by semi-nested polymerase chain reaction, the bonds of DNA were observed in electrophoresis with 2% gel agarose.

 Results

 250 patients were studied 66.9% of whom were positive and 33.1% negative for TTV. TTV-PCR results were also studied in 250 blood donors 41% of whom were positive and 59% negative. There was a significant difference (p=0.0001) between patients and the control group.

 Conclusions

 TTV has a high prevalence in recipients and blood donors. Blood transfusion probably is not the only way for transmission of TTV, and other ways such as oral-fecal route can also play a role for transmission of TTV.

 Key words : TTV, Blood donor, Blood recipient

  Abstract

 Background and Objectives

 The incidence of post transfusion hepatitis has been reduced by blood donor screening for HBsAg, but the HBV infection is still responsible for certain cases of post-transfusion hepatitis world-wide. An estimate of the rate of HBV DNA and anti-HBc positive units is important for evaluation of the need for anti-HBc blood donor screening. In this study, the HBsAg negative blood units were evaluated for anti-HBc and all of anti-HBc positive units were tested for HBV DNA by PCR method.

 Materials and Methods

 Extra samples were collected from 2000 HBsAg, anti-HCV, anti-HIV and RPR-negative blood donors. All of the samples were examined by the approved anti-HBc assay. All anti-HBc positive samples were tested by anti-HBs assays and evaluated for HBV DNA (PCR). The sensitivity of the HBV DNA (PCR) assasy was estimated to be 300 geq/ml according to VQC proficiency panels.

 Results

 230 (11.5%) out of 2000 samples were positive for anti-HBc. 179 (77.8%) out of 230 anti-HBc positive samples were HBsAb positive, and 51(23.2%) HBsAb negative. All 230 samples were assayed for single HBV DNA (PCR) 227 of which came out to be negative for HBV DNA (PCR). Three blood donors were recalled and new samples from two of whom were collected. These new samples were negative for HBV DNA.

 Conclusions

 Further study for evaluation of HBV DNA in anti-HBc positive blood units with full automatic instruments and usage of blood bags with accessories is strongly recommended.

  Abstract

 Background and Objectives

 Thrombophilia is a common and dangerous disease. It may be heritable or acquired. A substitution in the 3' untranslated region of prothrombin gene (PRT-G20210A) has been reported to be related to thrombophilia in Caucasians, but this relationship remains in debate in other populations. The hetrozygote form of PRT G20210A gene variant expresses prothrombin 25% higher than the average level. This study was performed to evaluate the prevalence of G20210A gene mutationas in thrombotic patients.

 Materials and Methods

 In this descriptive and cross sectional study, 299 patients with venous thromboembolism were selected. The subjects consisted of 116 male and 183 female. Traditional risk factors for venous thrombosis and pulmonary thromboembolism such as proteins C, S, ATIII deficiency and APC-resistance were investigated as well.

 Results

 We found that only 1.7% of patients with venous thromboembolism were carrier of prothrombin G20210A mutation which is one of the prevalent point mutations in Caucasians patients (18%). The homozygote form of PRT G20210A was not found.

 Conclusions

 Prothrombin G20210A mutations in Iranian population as opposed to western populations are rare (1.7%) with APC-resistance being common in the former.

 Key words: Prothrombin, Mutation, PCR, Venous thromboembolism

  Abstract

 Blood-letting is defined to be the withdrawal of blood from a patient. Considering the mysterious, life-saving, and occassionally miraculous nature of blood during the evolving history of man, civilization, and science, this red liquid being the token of life and death throughout centuries was used as evidence for clinical diagnosis of special diseases or otherwise as definite and soothing treatment of patients. Based on the existing evidence, hijama or blood withdrawal in cultural and religious beliefs and customs of certain tribes has had even a special status in saving man from devil or evil forces. The accessible old documents show the expansion of blood drawing as a known life-saving element and treatment method in ancient Egypt, Greece, and Rome in different forms including hijama. However, the development of medical sciences, particularly transfusion medicine and blood transfusion sciences, the treatment and preventive role of hijama and other methods like arteriotomy and leech cupping started to get less prominence except for some eastern countries especially Islamic states where hijama is still employed to relieve soul and treat diseases as a tool of preserving traditions. In Iran, considering the available standards based on which potential blood donors with recent hijama experience are defered for one year, it is necessary to raise awareness of all those involved in the field of blood transfusion and the whole community about the history of hijama so as to see how we can better deal with this historical and traditional controversial topic.

 Key words : Leeching, Venesection, phlebotomy, Blood letting

  Abstract

 Background and Objectives

 Umbilical cord is one of the major sources of mesenchymal stem cells. In this study, the differentiative potential of human umbilical vein mesenchymal stem cells into hepatocyte-like cells has been investigated by cellular uptake of indocyanine green.

 Materials and Methods

 Human umbilical vein mesenchymal stem cells ( UVMSCs) were incubated for 2 weeks in the medium containing hepatocyte growth factor they were also treated with the medium containing oncostatin M for another 2 weeks. The differentiated cells were analyzed by uptake of indocyanine green (ICG), immunofluorescence analysis, and periodic acid-Schiff (PAS) staining.

 Results

 The differentiating cells showed transition from bipolar fibroblast-like morphology to round or oval shape. Cellular uptake of ICG was detected in differentiated cells. Glycogen storage was determined by PAS staining in differentiated cells which were immunoreactive to albumin.

 Conclusions

 Based on these observations, we can conclude that UVMSCs are able to differentiate into hepatocyte-like cells in vitro and ICG-staining is a useful marker to identify differentiated hepatocytes from human umbilical vein mesenchymal stem cells.

 �

 Keywords : Cell Differentiation, Umbilical Veins, Mesenchymal Stem Cells, Hepatocytes, Indocyanine Green�

  In vitro differentiation of human umbilical vein

  mesenchymal stem cells into hepatocyte-like cells

  Raufi A.¹ , Amini A.¹ , Azadbakht M.¹ , Nikkhoo B.^2,3 , Aboozari M.³ , Fathi F^2,3

  ¹ Razi University, Kermanshah, Iran

  ² Cellular and Molecular Research Center, Kurdistan University of Medical Sciences, Sanandaj, Iran

  ³ Kurdistan University of Medical Sciences, Sanandaj, Iran

  Abstract

 Background and Objectives

 Umbilical vein mesenchymal stem cells (UVMSC) which are multipotent precursors have been recently isolated. They are capable of differentiating into various cell types including neural and cardiac cells. In this study, the differentiation ability of human UVMSC into hepatocyte cells has investigated with concentration on hepatic genes expression profile.

 Materials and Methods

 The UVMSC were isolated and cultured in differentiation medium containing hepatocyte growth factor (HGF) and oncostatin M (OSM) using two step protocol. Differentiation of UVMSCs into cells expressing liver-specific genes was investigated by RT-PCR and immunocytochemistry.

 Results

 The cells showed the remarkable transition from bipolar fibroblast-like morphology to epithelial and polygonal shapes. The temporal gene expression pattern of hepatocyte-specific genes such as CK8, CK18, HNF3β, c-Met, AFP, TTR, G6P, Transthyretin, and ALB were detected during differentiation. The immunoflourescent analysis also showed that the differentiated cells were stained positively for CK-18 protein.

 Conclusions

 Findings indicate that UVMSC has a potential for differentiation into the hepatic like cells.

 Key words : Umbilical Veins, Mesenchymal Stem Cells, Hepatocyte Growth Factor, Oncostatin M 

 Sci J Iran Blood Transfus Org 2011 8(2): 79-87

  Received: 29 Nov 2010

  Accepted: 15 Feb 2011