Use of antioxidant nanoparticles to reduce oxidative stress in blood storage.

Oxidative damage by free radicals has a negative effect on blood quality during storage. Antioxidant nanoparticles can prevent oxidative stress. We use SOD-CAT-Alb-PEG-PLGA- nanoparticles to reduce the effects of oxidative stress in blood storage. Electrospray was employed to prepare nanoparticles. Nanoparticles entered the test bags and were kept for 35 days from the time of donation under standard conditions. On target days, experiments were performed on the samples taken. The examination included blood smear, red blood cells count, hemoglobin, hematocrit, K, Fe, glutathione peroxidase, glutathion reductase, glucose-6-phosphate dehydrogenase, prooxidant-antioxidant balance, malondialdehyde, and flow cytometric assay for phosphatidylserine. The repeated measures analysis was performed on samples every week. Morphological changes were less in the test group compared to the control. The quantitative hemolysis profile test showed significant changes in the test and control groups (p < 0.05) in consecutive weeks except for K and Fe. Oxidative stress parameters too showed a significant change during the target days of the examination (p < 0.05). Also, the phosphatidylserine expression was increased in control groups more than test in consecutive weeks (p < 0.05). It seems that the use of antioxidant nanoparticles improves the quality of stored red blood cells and can prevent posttransfusion complications and blood loss by reducing oxidative stress.

Dose-dependent efficacy of antioxidant nanoparticles on red blood cells storage.

BACKGROUND: Transfusion of healthy red blood cells (RBCs) after storage is important. One of the storage lesions on blood bags is oxidative stress. One way to prevent increased oxidative stress is to use antioxidant nanoparticles (NPs). Superoxide dismutase (SOD) and catalase (CAT) play an important role in antioxidant defense on RBC. poly lactic-co-glycolic acid (PLGA) is a nontoxic biodegradable polymer that is approved by the Food and Drug Administration for drug delivery. This study aimed to assess dose-dependent efficacy of SOD-CAT-polyethylene glycol -PLGA on RBCs storage. MATERIALS AND METHODS: Using a descriptive study, during 1 month, twenty donors from Bojnourd Blood Donation Center were selected. NPs with different concentrations were injected into the satellite bags after directing blood to them. On target days, experiments were performed on the samples taken. Electrospray was employed to prepare SOD-CAT-PLGA NPs. Twenty packed RBCs were isolated from the whole blood bags by the mechanical method, and certain amount of product was transferred to the satellite bags. On days 1, 7, 14, 21, 28, and 35, bags were sampled. Malondialdehyde (MDA), prooxidant-antioxidant balance (PAB), and Annexin V were performed on the samples taken. The repeated measures analysis with the help of SPSS software version 20 was performed on samples. RESULTS: MDA increased in both groups. The maximum increase in test group was seen in concentration 12 mg (MDA Day 14, test [1.93 ± 0.3], [P MDA < 0.001]). Maximum increase in PAB was seen in concentration 12 mg (from 444 ± 1.7 to 563 ± 2.5) (P PAB = 0.000). Furthermore, PS expression increased in the concentration of 12 mg greater than other concentration in consecutive (from 5.00 ± 0.8 to 22.26 ± 1.7, [P < 0.001]). CONCLUSION: Evaluation of dose dependency showed that different concentrations of antioxidant NPs affect RBC. This effect can be changed oxidative stress and apoptosis. Using both changes to evaluate functional and toxicity can be helpful.

Adoption of Iran's code of ethics for blood donation and transfusion as a public health policy.

Blood is a public resource of human origin and its transfusion process is essential to individual and public health. This study aimed to develop a national code of ethics for blood donation and transfusion (BDT). This was a qualitative research with a multi methods approach in which a combination of methods including situational analysis, focus group discussion and expert panels were used. After situational analysis and orientation, the code of ethics for BDT was developed based on the findings of a content analysis within the framework of the four principles of biomedical ethics. The results were categorized into two sections: situational analysis and underpinnings measures, and the clauses of the code. The Iranian Blood Transfusion Organization has carried out three essential supportive measures over the past decades: approval of insurance coverage of blood recipients against communicable diseases; inclusion of 14 blood services in the book of "Relative Value Units of Health Services"; and formation of the National Ethics Committee of Transfusion Medicine. After recognition and orientation, the national code of ethics for BDT was adopted and imparted to blood donation centers. The code consists of two sections: "Blood Transfusion Centers: Donors and Donation" in 19 clauses, and "Hospitals: Patients" in 8 clauses. The national code of ethics for BDT establishes moral norms in order to protect the rights of blood donors and recipients. It could also serve as a basis for addressing the related ethical challenges and right decision-making in the area of BDT.

Association of ABO and Rh Blood Groups to Blood-Borne Infections among Blood Donors in Tehran-Iran.

BACKGROUND: The aim of this study was to investigate the prevalence of hepatitis B, hepatitis C, HIV and syphilis infections in blood donors referred to Tehran Blood Transfusion Center (TBTC), and determine any association between blood groups and blood- borne infections between the years of 2005 and 2011. METHODS: This was a retrospective study conducted at TBTC. All of the donor serum samples were screened for HBV, HCV, HIV and syphilis by using third generation ELISA kits and RPR test. Initial reactive samples were tested in duplicate. Confirmatory tests were performed on all repeatedly reactive donations. Blood group was determined by forward and reverse blood grouping. The results were subjected to chi square analysis for determination of statistical difference between the values among different categories according to SPSS program. RESULTS: Overall, 2031451 donor serum samples were collected in 2005-2011. Totally, 10451 were positive test for HBV, HCV, HIV and syphilis. The overall seroprevalence of HBV, HCV, HIV, and syphilis was 0.39%, 0.11%, 0.005%, and 0.010%, respectively. Hepatitis B and HIV infections were significantly associated with blood group of donors (P <0.05) ; percentage of HIV Ag/Ab was higher in donors who had blood group "A" and percentage of HBs Ag was lower in donors who had blood group O. There was no significant association between Hepatitis C and syphilis infections with ABO and Rh blood groups (P>0.05). CONCLUSION: Compared with neighboring countries and the international standards, prevalence of blood-borne infections is relatively low.

Evaluation of circulating natural type 1 interferon-producing cells in HIV/GBV-C and HIV/HCV coinfected patients: a preliminary study.

BACKGROUND: GB virus-C (GBV-C) is a flavivirus that probably influences HIV infection-associated disease among HIV/GBV-C coinfected patients and inhibits the progression of HIV infection to AIDS. To address the possibility of immune-modulating effects of GBV-C coinfection in HIV patients, we evaluated interferon-producing cells in HIV/GBV-C coinfected patients and compared them to HIV-infected patients. METHODS: We performed a pilot study to enumerate interferon-producing cell count by two-color flow cytometric analysis and also for determining the frequency of ongoing GBV-C and HCV infection among patients infected with HIV. Then, 83 asymptomatic HIV-positive patients were considered for evaluation of interferon-producing cells. Eighty three patients were stratified in four groups according to the HCV and GBV-C infection status: patients infected with HCV and GBV-C (GBV-C+/HCV+), patients infected with GBV-C but not with HCV (GBV-C+/HCV-), patients infected with HCV but not with GBV-C (GBV-C-/HCV+), and patients not infected by GBV-C and HCV (GBV-C-/HCV-). RESULTS: GBV-C was detected in plasma samples from 15.5% of HIV-infected patients and the frequency of HCV infection was 47.5%. Interferon-producing cells in GBV-C coinfected individuals revealed a wider range of numbers; however, there was no significant difference in the interferon-producing cell count among HIV-infected individuals. CONCLUSIONS: It does not appear that GBV-C coinfection affects generation or redistribution of interferon-producing cells in HIV-infected patients with relatively intact immune system.

Multidrug resistance inhibition by antisense oligonucleotide against MDR1/mRNA in P-glycoprotein expressing leukemic cells.

INTRODUCTION: Acute myeloblastic leukemia (AML) is the most common form of acute leukemia in adults. One major problem in this disease is the emergence of leukemic blast cells that are resistant to anticancer drugs. This phenomenon is termed multidrug resistance (MDR). One cause of MDR is the expression of the MDR1 gene and its product, P-glycoprotein (Pgp). AIM: In the present study, we tried to inhibit the MDR phenotype with MDR1/mRNA/Pgp in leukemic cells using different antisense sequences and two non-viral vectors. MATERIALS AND METHODS: The Pgp expressing cell line was established from a parental K562 (Erythroleukemia) cell line with increasing concentrations of doxorubicin, and named KDI/20. In order to reverse the MDR phenotype due to Pgp expression, four different sequences of sense, antisense and one random sequence with phosphorothioate (PTO) modification (PS-ODN) against MDR1/mRNA were synthesized. They were used on the KDI/20 cells in combination with two non-viral vectors: (1) Fugene 6 transfection reagent (cationic lipid) and (2) polyethylenimine (cationic polymer). The effect of PS-ODN was assessed at the cellular level by flow cytometry (for Pgp detection), and Rhodamine 123 assay (for functional assessment of Pgp) at the molecular level by RT-PCR (for MDR1/mRNA detection) and MTT assay in order to assess the sensitivity of cell to doxorubicin. RESULTS: The results showed a decrease in the percentage of Pgp protein and MDR1/mRNA expression and an increase in the accumulation of Rh123 and drug sensitivity of cells to doxorubicin by antisense I and III. The reduction of MDR1/mRNA was more significant than its protein reduction. Therefore, our data showed that antisense can reverse the MDR phenotype at the transcription level and the PEI vector is more efficient than cationic lipid.

Assessment of in vitro cytokine response in hemophilia A patients with or without factor VIII inhibitory antibody.

Factor VIII (FVIII) inhibitor antibodies are produced in a proportion of hemophilia A patients. Development of anti-FVIII inhibitor antibodies is a T cell-dependent response, mediated by FVIII specific CD4(+) T cells. This study was performed to investigate the contribution of T helper (Th) cell-mediated cytokine response in inhibitor production. Peripheral blood mononuclear cells (PBMCs) were obtained from hemophilia A patients with (n = 14) or without inhibitor (n = 14) and from normal individuals (n = 14). Following stimulation of PBMCs with rFVIII and phytohemagglutinin (PHA) mitogen, the secreted cytokines, interferon-gamma (IFN-gamma), interleukin-10 (IL-10), and transforming growth factor-beta1 (TGF-beta1), in culture supernatant and the proliferative response were assessed using sandwich ELISA and (3)H-thymidine incorporation, respectively. No significant proliferative response to FVIII was observed, whereas PHA induced a strong response in all groups. No cytokine secretion was observed in response to FVIII stimulation. Although PHA induced IL-10, TGF-beta1 and IFN-gamma secretion in all groups, the level of IFN-gamma was significantly lower in hemophilia A patients than in normal individuals (p < 0.0001). The levels of TGF-beta1 and IL-10 were similarly higher in patients compared with normal subjects, but the difference was not statistically significant. Lack of FVIII-induced proliferative response and cytokine production together with reduced secretion of PHA-induced IFN-gamma in both groups of patients suggest involvement of nonspecific immunosuppression possibly due to hepatitis C virus (HCV) infection observed in the majority of patients.

Relation of factor VIII and IX inhibitors with ABO blood groups in 150 patients with haemophilia A and B.

Many investigations have proved relations between ABO blood groups with some diseases and factor VIII and von willebrand level in plasma. In this study we investigated a relation between ABO blood groups and factor VIII and IX inhibitors in 102 patients with haemophilia A and 48 patients with haemophilia B. The assay of inhibitor was done by Bethesda method. There were no relation between ABO blood groups and factor VIII and IX inhibitors.