[Cytotoxicity of three species of centaurea genus on acute lymphoblastic leukemia cell line [Nalm-6]]

Background: Medicinal plants are considered as one of the ideal therapeutic sources for cancer prevention or treatment due to their bioactive contents and low side effects to humans. Centaurea genus have shown potential anti-tumor activity on some cancer cell lines in previous studies

Objective: The aim of this study was to evaluate the cytotoxic effect of three species of Centaurea genus [C. albonitens, C. pseudoscabiosa, C. salicifolia] on Nalm-6 cells

Methods: Nalm-6 cells were treated with increasing concentrations of the methanolic extracts of the aerial parts of three Centaurea species, then, their cytotoxic and anti-proliferative effects were evaluated using trypan blue, MTT and DAPI staining assays. Moreover, annexin V/PI staining and cell cycle analysis were performed for further investigation

Results: The results of trypan blue, MTT assay and DAPI staining revealed that all 3 extracts exhibit cytotoxic and anti-proliferative properties against Nalm-6 cells in a dose- and time-dependent manner [P>/=0.001]. Interestingly, there was no considerable cytotoxicity in normal cells, MDBK. The flow cytometric analysis validated that Centaurea dose dependently induces apoptosis and G1 phase arrest in Nalm-6 cells [P>/=0.01]

Conclusion: Due to the safety profile of the natural products, our study suggests that Centaurea extracts might provide insight into a novel therapeutic strategy and may confer advantages for acute lymphoblastic leukemia treatment, however further researches are necessary

Seroepidemiology of HBV infection in South-East of Iran; a population based study

Hepatitis B virus [HBV] infection is a major risk factor of cirrhosis and hepatocellular carcinoma affecting billions of people globally. Since information on its prevalence in general population is mandatory for formulating effective policies, this population based serological survey was conducted in Sistan and Baluchistan, where no previous epidemiological data were available. Using random cluster sampling 3989 healthy subjects were selected from 9 districts of Sistan and Baluchistan Province in southeastern Iran. The subjects' age ranged from 6 to 65 years old. Serum samples were tested for HBcAb, HBsAg. Screening tests were carried out by the third generation of ELISA. Various risk factors were recorded and multivariate analysis was performed. The prevalence of HBsAg and HBcAb in Sistan and Baluchistan was 3.38% [95% CI 2.85; 3.98] and 23.58% [95% CI 22.29; 24.93] respectively. We found 8 cases of positive anti-HDV antibody. Predictors of HBsAg or HBcAb in multivariate analysis were age, marital status and addiction. The rate of HBV infection in Sistan and Baluchistan was higher than other parts of Iran. Approxi-mately 25% of general population in this province had previous exposure to HBV and 3% were HBsAg carriers. Intrafamilial and addiction were major routes of HBV transmission in this province

Infection of MSCs by adenovirus expression system expressing NRF2 as a cytoprotective factor

Poor viability of Mesenchymal Stem Cells [MSCs] following transplantation is one of the major challenges in their therapeutic application. Manipulation of MSCs by the genetic engineering method is one of the strategies used to protect the cells against cytotoxic microenvironment. However, maintaining multi differentiation capacity of MSCs following manipulation is important. We investigated if the manipulation of MSCs with NRF2 affects the multi differentiation capacity. MSCs were isolated from bone marrow. NRF2 was isolated and TOPO cloned into the pENTR vector. The recombinant vector was transferred into pAD/CMV/V5-DEST vector by gateway technology. Recombinant adenovirus was produced in AD293 cells, followed by being infected into MSCs. Expression of NRF2 was verified by RT-PCR. The NRF2 engineered MSCs were exposed to stress conditions followed by the evaluation of the cells viability and apoptosis. Finally, NRF2 expressing MSCs differentiation into osteoblast and adipocyte lineages was studied. NRF2 was successfully expressed in MSCs. NRF2- MSCs differentiation into osteoblast and adipocyte lineages indicating overexpression of NRF2 does not affect the differentiation property of MSCs. Expression of NRF2, a well known cytoprotective factor, by using adenovirus expression system does not intervene in the differentiation capacity of MSCs. NRF2-MSCs might be applicable for stem cell-based cell therapy in future

[Comparison of cytokine growth factors in platelet supernatant, platelet lysate and activated platelet-rich plasma for therapeutic applications]

Background: platelet-Rich Plasma [PRP] is a concentration of human platelets in a small volume of plasma. It has been shown that effects of this product on various cell types is due to synergistic effects of some proteins of platelet alpha granules as platelet growth factors. The purpose of this study is definition of the more efficient platelet product, with higher protein and growth factor concentration for using in various applications

Materials and Methods: in this study, platelet-rich plasma was isolated by apheresis method. Activated Platelet-rich Plasma [AP] was prepared by platelet activation by thrombin and calcium chloride. Platelet Supernatant [PS] and Platelet Lysate [PL] preparation was performed by high speed centrifugation [900g], and freezing and thawing of platelet rich plasma, respectively. The growth factors and protein concentrations in these platelet products were measured by ELISA and Bradford methods, respectively

Results: platelet concentration of Platelet-rich Plasma was 1.066×10[9] +/- 0.15×10[9] /ml [Mean +/- SD]. According to the results, concentrations of PDGF, TGF-beta, FGF, EGF and protein were few in Platelet Supernatant [0.24 +/- 0.03 ng/ml, 4.5 +/- 0.4 ng/ml, 0.8 +/- 0.06 pg/ml, 2.6 +/- 0.3 pg/ml and 460 +/- 6 mg/ml, respectively] and significantly increased by platelet activation [23.0 +/- 2.1 ng/ml, 19 +/- 3.0 ng/ml, 7.4 +/- 1.0 pg/ml, 23.3 +/- 3.0 pg/ml and 480 +/- 4 mg/ml, respectively] or freezing and thawing of PRP [18.1 +/- 1.9 ng/ml, 16.0 +/- 3.0 ng/ml, 10.2 +/- 1.3 pg/ml, 31.0 +/- 4.0 pg/ml and 490 +/- 5 mg/ml, respectively] [P<0.05]. We showed that concentration of PDGF and TGF-beta as two major platelet growth factors is significantly higher than FGF and EGF in all of three platelet products [P<0.05]. We also found that FGF and EGF concentrations are significantly higher in Platelet Lysate [10.2 +/- 1.3 and 31.0 +/- 4.0] than Activated Platelet-rich Plasma [7.4 +/- 1.0 and 23.3 +/- 3.0] [P<0.05], and TGF-beta in Platelet Supernatant [4.5 +/- 0.4], in contrast to two other platelet products, is significantly higher than PDGF [0.24 +/- 0.03] [P<0.05]

Conclusions: preparation of Platelet-rich Plasma by apheresis method, gives a high platelet count. Protein extraction by PRP activation yields a higher concentration of two major factors [PDGF and TGF-beta], in spite of higher amounts of protein, FGF and EGF in Platelet Lysate. More studies are needed to define the relation between higher growth factor concentration or protein content and effects in various applications

[Comparison of cytokine growth factors in platelet supernatant, platelet lysate and activated platelet-rich plasma for therapeutic applications]

Platelet-Rich Plasma [PRP] is a concentration of human platelets in a small volume of plasma. It has been shown that effects of this product on various cell types is due to synergistic effects of some proteins of platelet alpha granules as platelet growth factors. The purpose of this study is definition of the more efficient platelet product, with higher protein and growth factor concentration for using in various applications. In this study, platelet-rich plasma was isolated by apheresis method. Activated Platelet-rich Plasma [AP] was prepared by platelet activation by thrombin and calcium chloride. Platelet Supernatant [PS] and Platelet Lysate [PL] preparation was performed by high speed centrifugation [900 g], and freezing and thawing of platelet rich plasma, respectively. The growth factors and protein concentrations in these platelet products were measured by ELISA and Bradford methods, respectively. Platelet concentration of Platelet-rich Plasma was 1.066x10[9] +/- 0.15x10[9] /ml [Mean +/- SD]. According to the results, concentrations of PDGF, TGF-beta, FGF, EGF and protein were few in Platelet Supernatant [0.24 +/- 0.03 ng/ml, 4.5 +/- 0.4 ng/ml, 0.8 +/- 0.06 pg/ml, 2.6 +/- 0.3 pg/ml and 460 +/- 6 mg/ml, respectively] and significantly increased by platelet activation [23.0 +/- 2.1 ng/ml,19 +/- 3.0 ng/ml,7.4 +/- 1.0 pg/ml,23.3 +/- 3.0 pg/ml and 480 +/- 4 mg/ml, respectively] or freezing and thawing of PRP [18.1 +/- 1.9 ng/ml, 16.0 +/- 3.0ng/ml,10.2 +/- 1.3pg/ml,31.0 +/- 4.0 pg/ml and 490 +/- 5 mg/ml, respectively], [P<0.05]. We showed that concentration of PDGF and TGF- beta as two major platelet growth factors is significantly higher than FGF and EGF in all of three platelet products [P<0.05]. We also found that FGF and EGF concentrations are significantly higher in Platelet Lysate [10.2 +/- 1.3 and 31.0 +/- 4.0] than Activated Platelet-rich Plasma [7.4 +/- 1.0 and 23.3 +/- 3.0] [P<0.05], and TGF-beta in Platelet Supernatant [4.5 +/- 0.4], in contrast to two other platelet products, is significantly higher than PDGF [0.24 +/- 0.03] [P<0.05]. Preparation of Platelet-rich Plasma by apheresis method, gives a high platelet count. Protein extraction by PRP activation yields a higher concentration of two major factors [PDGF and TGF-beta], in spite of higher amounts of protein, FGF and EGF in Platelet Lysate. More studies are needed to define the relation between higher growth factor concentration or protein content and effects in various applications

[Hijama through the passage of time]

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

[Seroepidemiology of the human Herpesvirus [HHV-8] in the haemodialysis patients blood donors and HIV-positive individuals in city of Tehran]

Human herpes virus 8 [HHV-8], also known as Kaposi sarcoma-associated herpes virus, is believed to be the infectious trigger for Kaposi sarcoma. HHV-8 transmission takes place via different routes such as saliva, sexual intercourse, mucosal contact and possibly blood transfusion. The objective of this study was to determine HHV-8 seroprevalence in otherwise healthy blood donors as immunocompetent hosts, in HIV positive individuals [immunocompromised hosts], and in hemodialysis patients as multi-transfused patients. This is the first time that research of this magnitude on HHV-8 prevalence is conducted in Iran. The study method was analytic-observational. We measured HHV-8 antibody levels in 118 hemodialysis patients, 35 HIV positive subjects and 256 healthy blood donors. The primary test method was ELISA; positive results were confirmed by IFA [immunofluorescence assay]. Subjects with positive results on both ELISA and IFA were regarded as HHV-8 cases. Overall, 20 hemodialysis patients [16.9%], 16 HIV individuals [45.7%] and 5 blood donors [2%] had HHV-8 antibodies. Analysis with ?2 tests did not show any significant association with sex [p=0.24], blood transfusion or the number of transfused blood units [p=0.36 and 0.73, respectively]. But there was positive correlation between age and the presence of antibodies [P=0.01]. Serologic prevalence of HHV-8 in blood donors [as apparently healthy individuals] proved to be lower than in other studies and, in some cases, equal to the figures from other countries. The high prevalence of HHV-8 antibodies in HIV positive individuals may be partly attributed to high-risk sexual behavior and repeated exposure to pathogenic agents. The higher prevalence of HHV-8 antibodies in hemodialysis patients as compared to blood donors [normal individuals] may be related to specific dialysis procedures or multiple transfusions with the resulting potential for infection

[Isolation and detection of RhD antigen from red blood cell [RBC] membrane by immune- precipitation method]

Rh [Rhesus] is a highly complex blood group system in man which plays an important role in transfusion medicine. The aim of this study was the isolation of RhD protein from the membrane of RBCs. In this experimental study immunoprecipitation method with human anti-RhD polyclonal antibody was utilized for the isolation of RhD antigen from Rh[+] human blood samples Proteins of RBCs were characterized by SDS-PAGE and Western blot analysis. Antigenicity of the RhD protein was assessed by ELISA using commercially available human anti-RhD polyclonal antibody with peroxidase conjugated goat anti-human as a secondary antibody. The results show that RhD protein has successfully been isolated by immunoprecipitation method. The expected size of RhD protein was confirmed by Western blot analysis. RhD antibody reacted with RhD antigen prepared from ghost with polyclonal antibody in ELISA, but no reaction was observed in Western blot analysis with monoclonal antibody: It is necessary to mention that this is the primary report of relative purification of RhD and further studies are recommended. The RhD may be helpful to further investigate the molecular basis of RhD protein and could be applicable for production of anti-D antibody in an animal model

[Assessment of Wilms tumor gene [WT1] in acute myeloid leukemia patients as a new marker to detect minimal residual disease [MRD]]

WT1 gene encodes a transcription factor that is involved in differentiation and proliferation of hematopoietic precursor cells as well as some other tissues like kidney, ovary, heart, etc. Quantitative assessment of WT1 gene expression is proposed as a useful marker in MRD detection and leukemia management. To assess the relevance of this gene, we analysed peripheral blood mononuclear cells of 62 AML patients [new cases] for the expression level of WT1 mRNA using Real-time quantitative RT-PCR. We followed the analysis up to 3 years, depending on patient availability. This study as a fundamental and applicable one was done cross-sectionnaly. Samples were obtained randomly from the AML patients referred to the BMT center, and selection was based on the diagnostic criteria defined by clinical wards. WT1 expression in MNCs of patients was compared with 24 healthy individuals [K562 cells considered to express WT1 gene equivalent to 10[6]]. Samples for diagnosis showed significantly high levels of WT1 expression [>80%]. After chemotherapy, its expression decreased [diminished about 1-2 log within induction therapy and around 3-4 log after consolidation therapy]. There was a noticeable correlation between the relative expression levels of WT1 and prediction of relapse [lower than gray zone versus higher than]. Patients whose WT1 expression levels remained lower than the gray zone benefit from a better compete remission. On the contrary, 1-6 months prior to overt clinical relapse in 6 patients, their WT1 expression raised to levels upper than gray zone. This study revealed that WT1 is a useful marker for detecting minimal residual disease, assessing chemotherapy effects, and predicting relapse in AML patients

Estimsation and comparison of the production cost of blood and blood products in 28 IBTO centers in 1381 [2002]

Estimation of the production cost of blood and blood products is essential to make more appropriate use of resources and to reduce costs. The production cost of each blood or blood product unit was estimated in 28 IBTO centers by taking into account provincial credits and officially communicated and provincial credits [with or without including credits of the first chapter]. The production cost of each product was multiplied in its own special index, depending on the complexity of production procedure. We did not use the industrial accounting method to estimate costs. The data obtained were analyzed by SPSS and Excel softwares. The production cost of one unit of blood/blood product in 71% of blood centers was estimated to be higher than the average country-wide cost provided from the provincial credits [with or without chapter 1 budget], and officially communicated and provincial credits. The same is true in 75% of blood centers whoes cost was provided from officially communicated and provincial credits without the first chapter being added. Most of IBTO centers spend more than the average country-wide cost to produce blood and blood products. It is recommended to revise resource management policies in blood transfusion centers

Need-assessment and research priorities-setting in IBTO Research Center

Setting research priorities in the cycle of research management is critical. The limitation in human and financial resources and policy changes are the most significant reasons necessitating research priorities to be set. Research prioritization can materialize and be effective at different levels ranging from macro and national to educational and research levels. To this end, IBTO Research and Education Deputy by this study has embarked on a serious measure in organizing and orienting investigations in IBTO.First the necessity of the implementation of the project was elaborated in the Research Council and priorities were set. Then, different procedures were conducted based on the guidelines of COHRED [Council on Health Research for Development] and by use of priority-setting instruments applied in research institutes. At the end, the results were reviewed by the Research Council so that the final priorities were approved of. In the present study, out of the whole number of forms distributed for priority-setting, blood centers, headquarter managers and consultants, and the faculty members had respectively a share of 64.28%, 33.33%, and 25.92% in responses. At the process of title collection, more responses were received as compared with the priority setting process. Finally, 99 research titles in 16 domains were approved of as final priorities by the IBTO Research Center. Priority-setting was conducted through the method recommended by COHRED for the first time in IBTO. In spite of the participation of the out-of-organization beneficiary, research centers, and scientific associations, the highest rate of participation goes to intra-organizational groups. Approved priorities can be implemented by a call for research, the creation of an evaluation system for recommended projects, and survey of approved projects. Thus, the most use can be made of financial and human resources for priority-setting

Seroepidemiologic HGV in blood donors haemodialysis patients, haemophiliacs and major beta thalassemics with history of liver disease

The prevalence of GBV-C and HGV in blood donor populations in developd countries based on HGV 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, heamodialysis patients, hemophiliacs, and beta thalassemics with a history of liver disease by Elisa technique. In this descriptive study, blood samples of 330 volunteer blood donors, 44 heamodialysis patients, 16 haemophiliacs, and 40 beta 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 HGV infection. For data analysis, Ch-square, Fisher exact test, and SPSS version 11.5 were used. This study showed that out of 330 healthy blood donors 14[4.2%], out of 44 heamodialysis patients 10[22.7%], out of 16 haemophiliacs 5 [30.3%] and out of 40 beta 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 heamodialysis patients, 4 [28.6%] of haemophiliacs and 9 [23.7%] of beta thalassemics. It was also found that 1 [8.3%] of heamodialysis patients, 1 [33.3%] of haemophiliacs, and 1 [50%] of beta thalassemics were infected with anti-HGV and HBsAg co-infection. The prevalence of HGV was high in multitransfused individuals including heamodialysis 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

case report of recurrent hydrops fetalis due to maternal alloimmunization with Kell antigen

During pregnancy, irregular blood group antibodies originating either from earlier pregnancies or from blood transfusions may severely affect child health. In this report, a case of maternal alloimmunization to Kell antigen is described.The mother had a history of partial mole and four repeated intrauterine fetal death due to hydrops fetalis.Screening of irregular blood group antibodies revealed that she has anti-Kell with the titer of 1:4096. Also in genetic analysis, a C677T homozygous mutation of MTHFR gene was found, which could potentially enhance destructive effects of anti-Kell antibody. The described case emphasizes the importance of being informed about the presence of irregular blood group antibodies during pregnancy which may cause recurrent hydrops

[ Evaluation of the attitude of Iranian women towards blood donation]

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. 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. 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