Correlation Between Mean Red Cell Transfusion Demand And Chelation Therapy In Multiply Transfused Thalassemia Major Patients.

Abstracts: Background & Objective: Life long red blood transfusion remains the main treatment for β thalassemia major patients. Transfusion-dependent patients, in the absence of chelation therapy, develop progressive accumulation of iron, which is responsible for tissue damage and, eventually, death. The factors, which influence the iron burden are type of chelation therapy and mean red cell transfusion requirement. Increasing red cell transfusion requirement, iron deposit and development of all antibodies complicates transfusion therapy in thalassemia patients. Aim is to investigate the patients for the red cell transfusion requirement compared on the basis of iron overload and type of chelation therapy. Methodology: Ninety eights patients were included in this study and samples collected and investigated for the red cell transfusion requirement, compared on the basis of iron overload and type of chelation therapy.Conclusion: Combination of two iron chelators (such as parenteral desferroxamine plus oral deferiprone) have been shown to produce additive and synergistic effects, may produce enhanced iron excretion, minimize side effects, decrease mean red cell transfusion requirement and improve compliance is strongly recommended in transfusion dependent thalassemia patients.

Prevalence and Incidence Of Transfusion Transmitted Infections Amongst VNRBDs In Central India.

Abstracts: Background: The aim of this study is to present the status of transfusion –transmitted infections among the apparently healthy donors so as to increase the awareness of complications of blood transfusion and make the clinicians more vigilant with regard to judicious use of blood. Methodology: A total of 15322 units of donor’s blood were screened from January 2008 to December 2010 at blood bank of C R Gardi Hospital and R D Gardi medical college. Results and Conclusion: The result of screening showed total seropositive samples for hepatitis B were 288 (1.88%), hepatitis C 52 (0.34%), human immunodeficiency virus 57 (HIV; 0.37% ) and Venereal Disease Research Laboratory Test 135 (VDRL; 0.88%) by using enzyme linked immunosorbent assay (ELISA ) methods, and rapid plasma regain (RPR) method for syphilis.

Multicenter evaluation of individual donor nucleic acid testing (NAT) for simultaneous detection of human immunodeficiency virus -1 & hepatitis B & C viruses in Indian blood donors.

BACKGROUND & OBJECTIVE: India has a high prevalence of HIV-1, hapatitis C and B virus (HCV and HBV) in the blood donors but has yet to implement nucleic acid testing (NAT) in blood screening. We undertook a multicentre evaluation of blood donor testing by NAT for simultaneous detection of HIV-1, HBV and HCV in a single tube and also to determine the feasibility of NAT implementation in India's low volume setting. METHODS: A total of 12,224 unlinked samples along with their serological results were obtained from representative eight blood banks in India and were individually manually tested by the Procleix Ultrio Assay (Chiron Corp. Emeryville, CA) for simultaneous detection of HIV-1, HCV, and HBV. RESULTS: Of the 12,224 samples tested, 209 (1.71%) were seroreactive. One hundred thirty three samples (1.09%) were reactive by Ultrio assay, 84 samples were seroreactive but NAT non reactive. There were eight NAT yield cases: 1 HIV, 1 HIV-HCV co-infection, and 6 HBV. INTERPRETATION & CONCLUSION: Our observed NAT yield for all three viruses was 1 in 1528 (0.065%). We estimate NAT could interdict 3272 infectious donations a year among our approximate 5 million annual donations.

Knowledge about HIV-AIDS among first-time and regular voluntary non-remunerated blood donors.

Acquired Immune Deficiency Syndrome (AIDS) is one of the serious public health problems in India. AIDS education has been considered as one of the main intervention for control. Sexual route is the major route of transmission of Human Immunodeficiency Virus (HIV); however, approximately 2.5% is transmitted through blood and blood products. The present study was carried out to know the level of awareness about HIV infection and blood donation among first time (190) and repeat (310) voluntary donors of all age groups. One pre-structured questionnaire was circulated among altruistic blood donors. About 96.6% donors want to become repeat donors. Majority of the donors had good knowledge about routes of HIV transmission. According to 97.4% donors, it is transmitted by sexual route, according to 87.4% of donors by sharing needle, according to 85% of donors by blood transfusion and 82.4% of donors believe through vertical transmission. However, 32.4% of the donors, still believe that HIV infection could be transmitted through blood donation. Intense motivational program among donors is needed to remove this myth. Regular donors were convinced the importance of regular and repeat blood donation. They came forward to donate blood for the cause of humanity (80.6%) and the sense of pride (27.79%). First time donors were less motivated by the cause of humanity (56.21%) and volunteered because of peer pressure (26.03%) and motivated by relative or friend. Donors were very alert about precaution to be taken for protecting themselves from danger of HIV infection and priority wise use of safe sexual practice (90%), disposable needles (61.43%) and receive tested blood (45.71%) whenever required. When in need of blood for relatives the donors will give priority to the quality (64.65%) and properly tested blood from voluntary blood donors (86.7%).

Effect of pre-storage gamma irradiation on red blood cells.

BACKGROUND & OBJECTIVE: The irradiation of blood components has received increased attention due to increasing categories of patients eligible to receive such blood to prevent transfusion-associated graft versus host disease. Irradiation leads to enhancement of storage lesions, which could have deleterious effects when such blood is transfused. The aim of the present study was to assess the biochemical changes during conventional preservation of irradiated and non-irradiated whole blood. METHODS: Ten units of whole blood were taken from healthy donors and divided into two parts. One aliquot was subjected to gamma irradiation and then stored under conventional blood banking conditions. Sampling was done from these irradiated and non-irradiated blood bags and tests for free plasma haemoglobin, plasma potassium and lactate dehydrogenase (LDH) were performed. RESULTS: A progressive increase in the mean values of plasma Hb, K+ and LDH was seen in both the groups. The increase was statistically significant. INTERPRETATION & CONCLUSION: Our findings indicated that the gamma irradiation of blood resulted in increased plasma haemoglobin, potassium and LDH. These biochemical changes might not have clinical significance when irradiated blood is transfused to a select group of patients. There is a need for further in vivo studies to follow up the consequences of transfusion of irradiated blood in patients.

Transmission of filarial infection through blood transfusion.

Microfilariae can be transmitted by blood transfusion and they may be circulated in the recipient's blood but they do not develop into adult worms. Mortality associated with transfusion associated filarial infection is not documented but it may give rise to morbidity in transfusion recipients in terms of allergic reaction. The present study was carried out to investigate the association of post transfusion reactions and filarial infections in an endemic area. About 11,752 transfusion recipients were followed up and in 15 months period, 47 (0.4%) post transfusion reactions (PTR) were reported. Routine investigations for post transfusion reaction were carried out in all 47 patients and their respective blood donor. Moreover, blood culture, microfilaria detection by concentration technique, filarial antibody and antigen detection (both by ELISA) were done in all subjects. Out of 47 patients showing post transfusion reaction, 29 (61.7%) patients developed allergic reaction. Eighteen (38.3%) patients having allergic reaction did not have previous history of blood transfusion and 14 (29.8%) of them received transfusion from blood donors who was either positive for microfilaria, filarial antigen or antibody. Microfilaremia was demonstrated in 4 (8.5%) patients and 5 (10.6%) blood donors. Microfilaria was concurrently present in 2 patients and their respective donors. Filarial antibody was detected in 27 (56.5%) patients and 26 (55.3%) blood donors but microfilaria was detected in 3 (6.4%) and 4 (8.5%) subjects, respectively. Antigen detection test correlated with microfileraemic state of subjects. The result shows that transfusion associated filarial infection may be a probable cause for transfusion-associated morbidity in endemic areas. In 14 (29.8%) patients having allergic reactions, the probable cause was transfusion-associated filarial infection. Filarial antigen detection test was found to be more useful in detecting infections. Blood donors with active history of filarial infection should be deferred from donating blood. Filarial antigen detection test may be employed as screening test for blood donors, if possible.

Cord blood from collection to expansion: feasibility in a regional blood bank.

This article reviews the various aspect of the experimental phase preceding the establishment of an umbilical cord blood (UCB) bank within a regular blood bank, a situation totally different from that of de novo establishing a cord blood bank having human and financial resources. An ethically approved two-year study has been conducted to determine the technical feasibility, and the practical problems that might be encountered such as public compliance, the additional workload, introduction of new activities ranging from collection and processing to progenitor expansion, infectious disease testing, development of a quality control system, record keeping and documentation, development of specific procedures and definitions of requirements. The cost benefit aspect, which will ultimately depend on the frequency of units release, was not considered in this study.

Transfusion transmitted diseases.

Transfusion transmitted disease (TTD) is a major challenge to the transfusion services all over the world. The problem of TTD is directly proportionate to the prevalence of the infection in the blood donor community. In India, hepatitis B/C, HIV, malaria, syphilis, cytomegalo virus, parvo-virus B-19 and bacterial infections are important causes of concern. Hepatitis B and C infections are prevalent in India and carrier rate is about 1-5% and 1%, respectively. Post transfusion hepatitis B/C is a major problem in India (about 10%) because of low viraemia and mutant strain undetectable by routine ELISA. HIV prevalence among blood donors is different in various parts of the country. It may not be so alarming as projected by some agencies. In one study from north India, confirmed HIV positivity was found in 0.2/1000 blood donor. Post transfusion CMV is difficult to prevent but use of leukocyte filters may help to reduce it significantly. Parvo virus B-19 infection in blood donors is 39.9% which may increase morbidity in multitransfused or immunocompromised patients. Current symphilis tests may not be sensitive but it should be continued to exclude high-risk donors. Malaria is a real problem for India due to the lack of a simple and sensitive screening test. Incidence of bacterial contamination is greatly reduced due to improved collection/preservation techniques and use of antibiotics in patients. However, proper vigilance and quality control is needed to prevent this problem. Total dependence of altruistic repeat voluntary donors and use of sensitive laboratory tests may help Indian blood transfusion services to reduce incidences of TTDs.

Evidence for clustered mannose as a new ligand for hyaluronan- binding protein (HABP1) from human fibroblasts.

We have earlier reported that overexpression of the gene encoding human hyaluronan-binding protein (HABP1) is functionally active, as it binds specifically with hyaluronan (HA). In this communication, we confirm the collapse of the filamentous and branched structure of HA by interaction with increasing concentrations of recombinant-HABP1 (rHABP1). HA is the reported ligand of rHABP1. Here, we show the affinity of rHABP1 towards D-mannosylated albumin (DMA) by overlay assay and purification using a DMA affinity column. Our data suggests that DMA is another ligand for HABP1. Furthermore, we have observed that DMA inhibits the binding of HA in a concentration-dependent manner, suggesting its multiligand affinity amongst carbohydrates. rHABP1 shows differential affinity towards HA and DMA which depends on pH and ionic strength. These data suggest that affinity of rHABP1 towards different ligands is regulated by the microenvironment.

Serological monitoring of thalassaemia major patients for transfusion associated viral infections.

A total of 39 patients with thalassaemia major who received multiple blood transfusions were followed up clinically and serologically for 3 successive years (1993, 1994, 1995). They were screened for hepatitis B surface antigen (HBsAg), and antibodies to hepatitis B core (HBc-total), hepatitis C virus (HCV), human immunodeficiency virus I and II (HIV-I/II) and cytomegalovirus (CMV-total). In spite of transfusing HBsAg screened (by third generation ELISA) blood from voluntary non-remunerated donors, there was a significant increase of HBsAg positivity (P < 0.001) from 17.9 per cent (1993) to 35.9 per cent (1994) to 69.2 per cent (1995). This was probably due to the prevalence of undetectable HBV infection in the population. Anti HBc was present in 17 (43.6%), 14 (35.9%) and 16 (41%) patients in consecutive years. An increase in the units of blood transfused was observed every year. Blood units were not screened for anti HCV antibodies but a gradual increase in positivity [9 (23%), 12 (30.7%) and 14 (35.9%) patients] was seen in consecutive years. Anti-HIV antibodies were found in a 16 yr old male who was included in the study without any clinical evidence of AIDS. Anti CMV antibody was found in 30 (76.9%), 32 (82%) and 29 (74.3%) patients without any apparent clinical infection. Some patients showed change of antibody pattern (from negative to positive or vice versa) and a few patients showed inconsistent results probably due to immune modulation. Recruitment of 'repeat' non-remunerated voluntary blood donors may reduce the risk of high HBV transmission.

The prevalence of transfusion transmitted diseases in renal transplant recipients.

Majority of renal transplant patients have history of blood transfusion. Out of a total of 120 renal transplant patients, 20 (16.6%) patients showed raised alanine aminotransferase (ALT) levels (> 45 IU) on 2 or more occasions at 2 weeks interval. This study was undertaken to estimate the risk of transfusion transmitted diseases (TTD) in these 20 renal transplant patients. Another 200 voluntary blood donors were also included as control. Both subject groups were screened by ELISA for HBsAg, anti HBc (IgG & IgM), anti HCV, anti CMV (IgM), anti HIV and VDRL tests for syphilis. A total of 11 (55%) patients and 3 (1.5%) blood donors showed the evidence of HBsAg positivity (p < 0.01). Anti HBc antibody was present in 12 (60%) patients and 39 (19.5%) donors which was significantly (p < 0.01) different. Anti HCV antibody was detected in 5 patients and in one blood donor (p < 0.01). However, anti CMV antibody was present in 4 (20%) patients and 3 (1.5%) donors, respectively. There was no evidence of HIV and syphilis infection in both these groups. High incidence of hepatitis markers were observed in renal transplant patients.

Effectiveness of mandatory transmissible diseases screening in Indian blood donors.

This study was undertaken to determine the prevalence of transfusion transmitted diseases (TTDs) among local blood donors, the safety offered by the four mandatory tests (for HIV, HBsAg, syphilis and malaria) and to assess alanine aminotransferase (ALT) as a surrogate test. A total of 313 blood donors were tested for HBsAg, hepatitis B core (HBc) antibody, hepatitis C (HCV) antibody, HIV antibody, and IgM antibody to cytomegalovirus (CMV-IgM). The serum alanine aminotransferase levels were also done on each unit of blood. The prevalence of various markers was 7(2.2%) for HBsAg, 57 (18.2%) for anti HBc (total), 1 (0.3%) for anti HCV, 16 (5.1%) for anti CMV. None of the donors were positive for HIV, VDRL or malaria. ALT level was raised in 16.5 per cent of donors and showed no correlation with hepatitis markers. ALT was not found to be useful as a surrogate marker for routine screening of donors. Sensitive tests like ELISA and immunofluoresence for malaria antigen should be applied for screening for malaria. VDRL test may be used to detect high risk donors rather than detection of syphilis when stored blood is used. HBsAg and HIV tests should be routinely done on every unit of blood and anti HCV tests should be done regularly, if possible.

Overexpression of Escherichia coli single stranded DNA binding protein under bacteriophage T 7 promoter.

A recombinant vector for overproduction of the E. coli single stranded DNA binding protein (E. coli SSBP) has been constructed. An E. coli strain carrying this plasmid produces up to 150 mg pure SSBP per litre of bacterial culture in a laboratory shake flask. Electron microscopy of the single stranded DNA complexed with SSBP shows characteristic "beaded string"-like appearance. Strong clustering of protein molecules on ssDNA is indicative of a highly cooperative binding.

Evaluation of haemotherapy in thalassaemias (20 years of Indian experience).

Two hundred fifty-one patients of beta-thalassaemia ranging from 3 months to 15 years of age were evaluated. They were maintained on hypertransfusion regimen and received periodic transfusions of group specific packed red cells. These multiple transfused patients were subjected to the tests for detection of transfusion malaria, HBsAg and allo-antibodies against red cells. Malaria infection and HBsAg were detected in 6.4% and 15.5% of patients respectively, while allo-antibodies were detected in 15.5% of patients. Thirteen patients (5.18%) developed hypersplenism and associated pressure symptoms due to splenomegaly for which they underwent splenectomy. Postsplenectomy period was uneventful and showed marked decrease in the frequency and quantitative requirements of transfusions and overall improvement in health. The continuing steady improvement of the prognosis in thalassaemia secondary to hypertransfusion regimen required us to transfer attention to other problems involved in thalassaemia management such as problems of hypersplenism and problems of multiple transfusion.

Plasmodial antigen detection by monoclonal antibody as a screening procedure for blood donors in transfusion medicine.

Very little information is available as regards the methods to be advocated to prevent transfusion malaria, especially in endemic countries. Most of the malaria non-endemic countries follow the rule of donor deferral for 3 years after malaria infection. This criterion cannot be followed in endemic areas since the majority of the population is continuously exposed to this infection. Therefore, there was a long felt need for a suitable screening procedure for blood donors to make the transfusion therapy safe. A total of 6,435 blood donors and 3,621 patients who received blood from these donors were studied by blood smears examination and malarial antigen detection by specific monoclonal antibody. Smear examination by Giemsa and acridine orange staining methods showed poor results (0.06% and 0.1% positivity respectively), probably due to low concentration of parasites. However, antigen detection by monoclonal antibody confirmed specific diagnosis in majority of these subjects. Blood smear examination failed to reveal malaria infection in 92.3% of antigen positive blood donors. It is, therefore, recommended that antigen detection by monoclonal antibody should be adopted as a routine screening procedure by the blood transfusion services in malaria endemic countries like India.