In-line leukoreduction filters: A new source of microparticle for human and animal study.

INTRODUCTION: The isolation of microparticles (MPs) from leukoreduction filters (LRFs) during cell extraction process introduced LRFs as a precious source of MPs for animal and human study. METHOD: LRFs were collected from Tehran Blood Transfusion Center. The back-flushing method was used for leukocyte extraction from the LRFs. MPs were isolated through double-step centrifugation. Dynamic light scattering (DLS), electron microscopy (EM), and flow cytometry were performed for the evaluation of MPs size, morphology, and structural properties respectively. Statistical analyses were carried out to evaluation of differences between test and control groups. a p-value less than 0.05 indicates significant differences. RESULT: DLS analysis showed that the average MP size in the test and control groups was 654.83 nm and 233.68 nm respectively. SEM images showed the spherical, oval, cell fragment, and micro-aggregate particles and TEM images demonstrated the mitochondrial-like body in the MPs. Flow cytometry studies also showed a significant increase in the percent of CD41, and CD14, and a significant decrease in the percent of CD235a in the test group compared to control (P value=0.029, P value=0.035, P value= 0.001 respectively). Moreover, the percentage of CD34 MPs indicated a borderline difference between the two groups (P value= 0.075). Finally count of MPs in the test and control groups was 1202095.34 and 280948.64, respectively and the difference was significant (P value=0.008). CONCLUSION: It is concluded that LRFs are a potential source of the large volume of various cell MPs with different phenotypical and structural properties for animal and human phase studies. Moreover, the investigation of LRFs as a source of different types of exosomes can shed new light on extracellular vesicle studies.

Quantification of hepatitis C virus in viremic blood donors in Iran: Need to reinforce post-donation follow up.

INTRODUCTION: Hepatitis C virus (HCV) infection is a public health problem and a major cause of chronic liver disease around the world. The main route of HCV transmission is contact with small quantities of infectious blood. Knowledge of the distribution of HCV viral load is essential to control HCV infection. This study aimed to investigate the HCV viral load distribution among Iranian blood donors. MATERIALS AND METHODS: This cross-sectional study was conducted on 160 HCV confirmed blood donors with detectable HCV RNA who referred to blood transfusion centers for post-donation counseling all over the country. HCV RNA was quantified using an in-house one-step real-time reverse transcription-polymerase chain reaction (RT-PCR) kit. Statistical analysis was performed in STATA version 13. RESULTS: The mean age of the participants was 37.66 years. Out of 160 subjects, 156 (97.5 %) were male. The median viral load of the subjects was 7.7 × 104 (range: 2.28 × 10 3-3.42 × 107 IU/mL). Out of 160 blood donors, 70 (43.75 %, 95 % CI 0.36-0.51) had a viral load ≤5 × 104 IU/mL, and 90 (56.25 %, 95 % CI: 0.49-0.64) had a viral load >5 × 104 IU/mL. DISCUSSION: The distribution of HCV viral load among viremic blood donors emphasizes on the role of post-donation follow up in identification of blood donors potentially need for HCV anti-viral therapies.

MicroRNA-21 over expression in umbilical cord blood hematopoietic stem progenitor cells by leukemia microvesicles.

Microvesicles are able to induce the cell of origin's phenotype in a target cell. MicroRNA-21, as an oncomir, is up-regulated in almost all cancer types such as leukemia which results in cell proliferation. In this study, we examine the ability of leukemia microvesicles to induce proliferation in hematopoietic stem progenitor cells (HSPCs) via microRNA-21 dysregulation. Herein, leukemia microvesicles were isolated from HL-60 and NB-4 cell lines by ultracentrifugation, and then their protein content was measured. Normal HSPCs were isolated from umbilical cord blood samples by a CD-34 antibody. These cells were treated with 20 and 40 µg/mL leukemia microvesicles for 5 and 10 days, respectively. Cell count, CD-34 analysis, and a microRNA-21 gene expression assay were done at days 5 and 10. HSPCs showed a significant increase in both microRNA-21 gene expression and cell count after treating with leukemia microvesicles compared with the control group. CD-34 analysis as stemness proof did not show any difference among the studied groups. This data suggests that HSPC proliferation followed by microRNA-21 gene over expression can be another evidence of a leukemia-like phenotype induction in a healthy target cell by leukemia microvesicles.

Leukemia cell microvesicles promote survival in umbilical cord blood hematopoietic stem cells.

Microvesicles can transfer their contents, proteins and RNA, to target cells and thereby transform them. This may induce apoptosis or survival depending on cell origin and the target cell. In this study, we investigate the effect of leukemic cell microvesicles on umbilical cord blood hematopoietic stem cells to seek evidence of apoptosis or cell survival. Microvesicles were isolated from both healthy donor bone marrow samples and Jurkat cells by ultra-centrifugation and were added to hematopoietic stem cells sorted from umbilical cord blood samples by magnetic associated cell sorting (MACS) technique. After 7 days, cell count, cell viability, flow cytometry analysis for hematopoietic stem cell markers and qPCR for P53 gene expression were performed. The results showed higher cell number, higher cell viability rate and lower P53 gene expression in leukemia group in comparison with normal and control groups. Also, CD34 expression as the most important hematopoietic stem cell marker, did not change during the treatment and lineage differentiation was not observed. In conclusion, this study showed anti-apoptotic effect of leukemia cell derived microvesicles on umbilical cord blood hematopoietic stem cells.

Seroprevalence of Hepatitis B Virus Infection and Its Risk Factors in the West of Iran: A Population-based Study.

INTRODUCTION: Hepatitis B virus (HBV) infection is a serious global public health problem affecting billions of people globally. The lack of information of its seroprevalence among the general population is an obstacle for formulating effective policies to reduce the burden viral hepatitis. Therefore, this population based serological survey was conducted in Kurdistan province, where no epidemiological data was available to determine the prevalence and risk factors of HBV infection. METHODS: 1613 healthy subjects were selected from all districts of Kurdistan province (in the western of Iran) using random cluster sampling. The subjects' age ranged from 6 to 65 years old. Serum samples were tested for HBcAb, HBsAg and anti-HDV antibody. Screening tests were carried out by the third generation of ELISA. Various risk factors were recorded and multivariate analysis was performed. RESULTS: The prevalence of HBsAg and HBcAb in Kurdistan was before 0.80% (95% CI 0.44; 1.34) and 5.02% (95% CI 4.03; 6.17), respectively. None of HBsAg carriers had positive anti-HDV antibody. Predictors of HBsAg or HBcAb in multivariate analysis were: older age and marriage. We did not find any significant differences between males and females. CONCLUSION: Our population based study suggests that intrafamilial HBV transmission plays a major role in HBV transmission in Kurdistan province. Furthermore, approximately 5% of general population in this province has prior exposure to HBV and less than 1% is HBsAg carriers. However, we could not find any case of HDV infection among them.

The antitumor efficiency of combined electrochemotherapy and single dose irradiation on a breast cancer tumor model.

BACKGROUND: The aim of this study was to investigate the antitumor effectiveness of electrochemotherapy with cisplatin combined with suboptimal radiotherapy doses. Tumor radiosensitization was evaluated on large invasive ductal carcinoma tumors in Balb/C mice. MATERIALS AND METHODS: Tumors of an average volume of 630 mm(3) were treated with cisplatin, electric pulses, radiotherapy, electrochemotherapy, alone as well as in appropriate combinations. Tumors were irradiated with Cobalt-60 γ-rays at doses 3 Gy and 5 Gy in combination with electrochemotherapy using cisplatin. Controls included each of the treatments alone as well as the combination of the radiotherapy with electric pulses alone or with cisplatin alone. Antitumor effectiveness was evaluated by tumor growth delay, tumor-doubling time, inhibition ratio and the objective response rates. RESULTS: As anticipated, electrochemotherapy was more effective than the treatment with cisplatin alone or the application of the electric pulses alone. When treatments were combined with tumor irradiation at either 3 or 5 Gy, the combination with electrochemotherapy was more effective: at 5 Gy, 2 animals out of 8 were in complete remission 100 days later. In general the higher 5 Gy dose of γ-radiation was more effective than the lower one of 3 Gy. CONCLUSIONS: The results of our study demonstrate that irradiation doses, 3 Gy or 5 Gy, increase the antitumor effectiveness of electrochemotherapy with cisplatin on invasive ductal carcinoma tumors. Good antitumor results were achieved in experimental tumors with a size comparable to clinical lesions, demonstrating that this three-modality combined treatment is useful for the treatment of large lesions even at sub-optimal radiotherapy doses.

WITHDRAWN: Low dose ribavirin for treatment of HCV infected thalassemia major patients: New indications for combination therapy.

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Prevalence and trends of human immunodeficiency virus, hepatitis B virus, and hepatitis C virus among blood donors in Iran, 2004 through 2007.

BACKGROUND: Evaluation and monitoring the prevalence of transfusion-transmissible viral infections in blood donors is a valuable index of donor selection and blood safety. This study analyzed the trends of blood-borne infections among Iranian blood donations during 4 years. STUDY DESIGN AND METHODS: Viral screening results of 6,499,851 allogeneic donations from 2004 through 2007 were analyzed. All donations were screened for hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency virus (HIV), and syphilis. The prevalence of HBV, HCV, and HIV infections per 100,000 donations and 95% confidence interval was calculated. The p value was estimated by chi-square test. RESULTS: The prevalences of HBV, HCV, and HIV decreased during the 4-year study from 2004 through 2007. The overall prevalence was 0.56% for HBV, 0.004% for HIV, and 0.13% for HCV. There was a significant and impressive decrease in hepatitis B surface antigen prevalence from 0.73% in 2004 to 0.41% in 2007. The prevalence of HIV appeared to have decreased from 0.005% in 2004 to 0.004% in 2007 although the decrease was not significant. HCV prevalence showed a slight decline in blood donations from 0.14% in 2005 to 0.12% in 2007. CONCLUSION: The trends of transfusion-transmitted infection prevalence in Iranian blood donations suggest that most of the safety measures employed in recent years in Iran have been effective.