The Scientific Journal of Iranian Blood Transfusion
Year:2006 | Volume:3 | Issue:3|Papers Count:14

Background and Objectives: Drug resistance remains one of the most important clinical obstacles in the treatment of some cancers. This drug resistance referred to as Multidrug Resistance (MDR) induces cross-resistance to many chemotherapy agents such as anthracyclines, vinca alkaloides, epipodophyllotoxins and Taxol. MDR is most likely due to the reduction of drug accumulation with an energy-dependent drug efflux pump. This drug pump is a 170 kDa transmembrane glycoprotein (Pgp).Materials and Methods: We developed a resistance subline of K562 by stepwise increase in concentration of Doxorubicin, and Pgp expression was verified by flowcytometry and RT-PCR methods. Cross resistance of the resistant cell line to Etoposide, Vincristine and Taxol was analyzed by MTT assay. Results: IC50 (the level of drug concentration inhibiting 50% of cell growth) of Doxorubicin, Etoposide and Taxol of parental K562 came out to be 100ng/ml and it was 50 ng/ml for vincristine. IC50 levels of these drugs on resistant K562 were 500, 500, 450 and 450ng/ml. These drugs also displayed 5-, 5-, 4.5-, and 9- fold resistance respectively.Conclusions: According to the results, expression of Pgp confers MDR phenotype to the K562 cell line and makes it resistant to most of anticancer drugs including anthracyclines, vinca alkaloides, epipodophyllotoxins and taxans. This MDR phenotype is a major obstacle of cancer treatment and in recent years investigators are trying to reverse it by gene therapy.

Background and Objectives: The most important aim of all blood transfusion centers is to recruit blood donors from low risk groups of society for blood donation to be embarked on voluntarily and regularly. In the city of Yazd, women form only 5% of blood donors. The aim of this study was to assess barriers to blood donation by women and the level of knowledge, attitude and practice regarding blood donation in the city of Yazd. Materials and Methods: In this cross sectional study, 1602 women were selected by clustering sampling method and interviewed using a specially formatted questionnaire. Data were analyzed by ANOVA and t-test. Results: Level of knowledge was different across various groups. Illiterate women and housewives had the least level of knowledge. 15% of women had donated blood at least once in the past. The most important motivational factor was sense of moral duty with a spiritual reward and the most important barriers to blood donation were anemia, fear, lack of time, and difficulty in access to donation sites. A significant percentage of women had wrong beliefs about blood donation. There was a direct relationship between knowledge and practice (p=0.0). The number of blood donations was correlated with increase in level of knowledge (p=0.0).Conclusions: Increase in the level of knowledge of women and correction of wrong beliefs should be the topmost priority. Further studies are needed to determine whether addressing anemia and women's health can tend to increase the number of female blood donors.

Background and Objectives: Providing fetal calf serum (FCS) alternatives as cell culture supplements is an important field of research to compensate for the FCS supply shortage. This study focused on preparation of fetal calf serum alternatives and their effects on growth and secretion of hybridoma cell lines. Materials and Methods: Outdated human platelet units undergo extraction for its growth factors to be obtained. Human AB blood group plasma was also converted to serum and its growth effect was compared to FCS, hypoxanthine-thymidine (HT) and RPMI1640 as cell culture media and supplement. Cell growth indices were preliminary counting of cells , confluency as surface area of plates filled with cells, and titration of monoclonal anti-A and anti-B blood group antibodies collected from cultured mouse hybridoma cells. Statistical analysis including one sample t-test, logarithmic multiple regression curve fit, and factor analysis was done by SPSS v12 software. Results: The four nutritional supplements of (1) human serum AB (AB), (2) human platelet extract (PLT), (3) equal mixture of AB & PLT (ABP), and (4) fetal calf serum as cell culture were examined on mouse hybridoma anti-A and anti-B monoclonal antibody producer cell lines for cell growth indices and compared with the same indices on RPMI1640 media. The growthstimulating effects in descending order of values were (1) ABP5%, (2) FCS10%, (3) ABP10%, (4) AB5%, (5) AB10%, (6) PLT5%, (7) ABP20%, (8) PLT10%, (9) PLT20%, and (10) HT; but AB20% inhibited growth of mentioned hybridoma cell lines. The titer of anti- A and anti-B monoclonal antibodies produced by cultured hybridoma on 5 and 10 percent concentration of AB, PLT and ABP compared to FCS5-10% at descending order were (1) PLT5%, (2) PLT10%, ABP5%, ABP10%, AB10%, and (3) AB5%. Conclusions: In general FCS had the following effects on curves of cell growth: (1) the highest increase on slope of multiplication (ascending) phase, (2) the highest increase on slope of death (descending) phase, and (3) the lowest duration of stationary phase. Then, FCS can be appropriate for growth of cells at initial low cell count. Human serum AB, human platelet extract, and equal mixture of both at optimum concentrations (these supplements at high concentrations killed cells) compared to FCS showed (1) decreased slope of multiplication phase, (2) decreased slope of death phase, and (3) increased duration of stationary phase. Thus, AB and PLT may be suitable for continuous cell culture systems in which cell survival during longer times is required. Factor analysis was introduced as a model to evaluate kinetics of cell growth at different supplements.

Background and Objectives: 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. Materials and Methods: 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 106).Results: 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 noticable correlation between the relative expression levels of WT1 and predicition 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.  Conclusions: This study revealed that WT1 is a useful marker for detecting minimal residual disease, assessing chemotherapy effects, and prediciting relapse in AML patients.

Background and Objectives: Hemostasis and control of bleeding are important principles in surgery. Control of bleeding with normal procedures such as cauterization and ligation is sometimes hard and even impossible. Considering different effects of collagen including hemostasis acceleration, complete absorption, tissue repair, and flexibility and appropriate traction, it can help maintain hemostasis.Materials and Methods: In a randomized controlled clinical trial, 92 cases and 91 controls were compared. Horse collagen (Antema) was used in cases while routine procedures were implemented in controls. Patients were evaluated in 4 groups involving dental procedures, pilonidal sinus open surgery, hemorrhoidectomy, and laminectomy. Statistical indices such as mean, standard error, and statistical tests such as t-test and chi-square were used in the analysis. Results: Mean age was 29.48 ± 1.29 in the cases and 33.2 + 1.5 in the control group. Seventy four (36 cases, 38 controls) undergoing dental procedures, 61(31 cases, 30 controls) pilonidal sinus open surgeries, 31(18 cases, 18 controls) hemorrhoidectomies, and 12(7 cases, 5 control) laminectomies were investigated. Despite higher severity of bleeding in cases (p=0.011, r=0.247), the time of bleeding control (p<0.001) and the time of complete hemostasis(p<0.001) were lower in cases than in the control group.Conclusions: Antema is effective in hemostasis and it may be used as an adjunct material in hemostasis.  

Background and Objectives: Considering the increasing trend of blood use, one of the main issues in blood safety is appropriate blood administration encompassing the size of needle lumen, use of high- flow sets for transfusion, and blood warming. Materials and Methods: In this descriptive study, 2713 hospitalized patients from two hospitals receiving blood and blood components were evaluated for blood administration. Results: From 2713 recipients, 2386 (88%) received unwarmed blood, and 230(8.5%) warmed blood. Regarding the rest 97 (3.5%) no data were received. Out of the total number of recipients, 65 (2.3%) received blood with high flow rate through needle and 2585 with low rate. Regarding the rest 64 no data were received. 1804 patients (66.9%) were transfused with number 18 needle, 464 (17.1%) with number 20, 119 (4.3%) with number 22, 122 (4.4%) with number 16, and a few (7.8%) with different needle numbers. Conclusions: The needle sizes used for blood transfusion were appropriate. But high-flow sets for rapid transfusion and blood warming methods were inappropriate.

Background and Objectives: Hepatitis B virus (HBV) is one of the main causes of hepatitis, cirrhosis and hepatocellular carcinoma, and vaccination by anti-HBs is the only reliable method for its prevention. Since blood transfusion is one of the most important transmission routes of this virus, blood recipients may always be at its exposure. Considering the importance of vaccination in multitransfused patients, we decided to evaluate HBsAb level in them.Materials and Methods: In this cross sectional study with simple sampling, 83 patients with major thalassemia, 5 with hemophilia, and 5 with fanconi anemia, having been referred to Hajar Hospital of Shahrekord to receive blood and blood products in the year 2003 were evaluated for their level of HBsAb. The level of HBsAb was measured with Diasorin kit (sensitivity=95%, specificity=98%). Data were statistically analyzed with chi-square test. Results: Thirty patients (28.5%) were nonimmune, 11 (12.5%) were partially immune, and 52 (59%) were immune. Eighty eight patients had previous history of hepatitis B vaccination. Out of those who had received the last vaccination within the last 5 years, 82% were immune (p<0.05) and out of those who had it earlier, only 35.7% were immune.  Conclusions: Based on the results of this study, determination of immune status in multitransfused patients and their immunization are important and revaccination after a 5-year lapse in patients is recommended.

Background and Objectives: Thalassemia major patients should continuously receive blood to survive. Permanent blood injection will expose patients' immune system to a broad spectrum of new antigens located on the surface of injected RBCs. Continuous exposure to foreign antigens may provoke antibody production against them in the patients who lack those Ags. This phenomenon is termed alloimmunization and can cause problems for preparation of compatible blood for transfusion. Materials and Methods: 65 patients were studied in this research. At first, questionnaires eliciting information about age, race, date of blood injection, splenectomy, presence of any underlying diseases, and any certain drugs (in patients' records) were filled out by all patients. For alloantibody screening, patients' serum samples were tested by the panels prepared in IBTO. Phenotypes of RBCs for Ags of ABH and Rh (D, C, E, c, e) were also determined by relevant antisera. Results: Out of 65 patients, only one case (1.53%) had been allommunized. The age range of males and females were 13 ± 6 and 13 ± 5 respectively. All of the patients were of Lurish race. The first blood transfusion in all of them occurred at the age below 3. Seventeen subjects were splenectomized. Conclusions: The reasons for low prevalence of alloimmunization against RBCs in thalassemia major patients may pertain to similarity of patients and donors by descent, blood transfusion before the age 3, and splenectomy in a few patients. Identification of alloantibodies by using the panel cells was the best method to provide patients with antigen negative and compatible blood for transfusion.

Background and Objectives: In some emergencies, blood transfusion is required while there is no time for crossmatching, antibody screening, and ABO/Rh typing. O negative blood can be used when there is not sufficient time for crossmatch. Case: A 68-year-old woman was transferred to the hospital cause of trauma. Due to her hemorrhage and paleness, intravenous isotonic fluids followed by 1 unit of O negative, 2 units of B positive, and 2 units of AB positive packed RBCs were administered. Her blood group was determined to be AB positive. She had neither any hemolytic symptoms nor any other problems and was discharged from the hospital. Conclusions: In emergencies when there is not enough time for crossmatching and antibody screening, O negative blood can be administered.