Regulated upon activation, normal T cells expressed and secreted (CCL5) in platelet concentrate: Role of mode of preparation and duration of storage.

BACKGROUND AND OBJECTIVES: Platelet concentrates (PCs) can be prepared in several different ways, and they can be stored over few days before the use. Regulated on activation, normal T cells expressed and secreted (RANTES) levels in these concentrates may vary depending on the type of preparation and duration of storage of this component. We measured RANTES levels in platelet supernatants in different preparations and with different storage duration. MATERIALS AND METHODS: Fifteen PCs were prepared by platelet-rich plasma (PRP) and buffy coat (BC) method each. Forty-two single donor platelets (SDPs) were prepared using cell separators Cobe Spectra, Trima Accel, and Amicus. Filtered PCs were prepared using labside and bedside filters. The supernatants were collected after 1, 18, 65, and 112 h of preparation. SDP samples were taken on the 0 day, 3rd day, and 5th day. In filtered PC, pre- and post-filtration samples were taken, and aliquots were frozen at - 56°C for the measurement of RANTES. RESULTS: RANTES at 1 h was 1210 ± 560 pg/ml in PRP-PC, 1384 ± 463 pg/ml in BC-PC. At 112 h, 1617 ± 451 pg/ml and 1949 ± 134 pg/ml, respectively. In SDP, 0-day level was 1850 ± 278 pg/ml and >2000 pg/ml on 5th day. In prestorage, filtered PC RANTES was 1035 ± 496 pg/ml, and in the poststorage sample, it was 310 ± 508 pg/ml. With bedside filters, presample showed 1243 ± 832 pg/ml and postsample showed 556 ± 748 pg/ml. CONCLUSION: The concentration of RANTES increased continuously from 1 h to 5 days of storage in all PCs. After 65 h, BC-PC showed higher levels of RANTES compared to PRP-PC. Filtered PRP-PCs appear to be the best in terms of low RANTES to prevent allergic reactions and cultures negative.

Blood Component Utilization for Disseminated Intravascular Coagulation (DIC) Cases with Respect to Underlying Condition.

To understand transfusion requirement in DIC with respect to (wrt) underlying cause. Patients' details were analyzed using parameters like yearly requirement, hemoglobin (Hb) and blood components utilization wrt age, sex and cause of DIC. Total 1931 DIC cases from 167 hospitals received 21,153 blood units including whole blood, red cell concentrate fresh frozen plasma, random donor platelets, single donor platelets and cryoprecipitate from year 2001 to 2013. Significant risk of DIC was for females compared to males (p < 0.0001). Whole blood utilization was 25.5 % in 2001 which reduced to 1.2 % in 2013. Mean ± SD Hb concentration was 8.6 ± 3.0 in females and 9.8 ± 3.2 g/dL in males. The lowest mean Hb was 7 g/dL in female DIC cases having malignancy. Septicemia was the major cause of DIC followed by obstetric complications. The highest requirement was for trauma associated with septicemia, requiring 35 units/case. Combination of two or more underlying causes for DIC increased the blood component requirement.

Time Dependent Release of Interleukin-8 and Tumor Necrosis Factor-α in Platelet Concentrate.

Contaminating white blood cells in stored platelet concentrate (PC) are the source of many pro-inflammatory cytokines. These are implicated in transfusion reactions. To study the release of interleukin (IL)-8 and tumor necrosis factor alpha (TNF-α) at different time interval in PC prepared by-platelet rich plasma (PRP) and buffy coat (BC) using different principles. Fifteen PCs were prepared by both the methods. The supernatants of PCs prepared by PRP and BC methods were collected aseptically after 1, 18, 65 and 112 h of preparation. pH, platelet and WBC counts were done. The supernatants were frozen in aliquots at -56 °C for measurement of IL-8 and TNF-α concentration using ELISA. The Mean ± SD value of WBC in PRP-PC was 7.4 ± 3.75 × 10(7) and in BC-PC 3.9 ± 2.2 × 10(7). The mean platelet counts were 6.05 ± 1.94 × 10(10) and 6.54 ± 2.18 × 10(10) respectively. The highest level of IL-8 in one hour was up to 30 pg/ml in both the type of PC. It increased up to 986 pg/ml in PRP-PC and 481 pg/ml in BC-PC at 112 h. IL-8 increased significantly during storage period of 5 days in both types of PCs (P0.000 and P0.01). TNF-α level remained low up to 18 h. The highest level was 72 pg/ml in PRP-PC and 57 pg/ml in BC-PC at 65 h. IL-8 levels significantly increased after one hour of storage and TNF-α. levels were low up to 18 h and then showed increase. The BC-PC had significantly low levels of IL-8 compared to PRP-PC (P0.0001).

Blood transfusion practice in obstetric and gynecology: impact of educational programs to create awareness for judicious use of blood components.

The study presents the data analysis (1) To find out the trend of blood component use during the period 2003-2010 and to determine impact of component awareness programs on reduction in whole blood (WB) and single unit transfusions. (2) To determine Hb trigger. The details about blood units issued were entered in the integrated blood bank management software as well as in Microsoft Excel. The data of 4,838 cases of pregnancy anemia; 2,244 receiving blood for obstetric (Ob) hemorrhage including 270 cases of disseminated intravascular coagulation; 1,413 women having Gynecological (Gy) bleeding; 911 Ob, 2,032 Gy and 740 surgeries for Gy malignancy were analyzed. During the years 2003-2010 there was gradual increase in component utilization for pregnancy anemia, Ob/Gy surgeries and Ob/Gy bleeding and significant reduction in WB transfusions due to component awareness programs. But single unit transfusions showed comparatively lower trend of reduction. The mean Hb was 6.4 g/dL for pregnancy anemia, 8.1 g/dL for surgeries and 7.3 g/dL for Ob/Gy bleeding.

Molecular characterization of ß-thalassemia in four communities in South Gujarat--codon 30 (G → A) a predominant mutation in the Kachhiya Patel community.

Different thalassemia mutations have been reported in various ethnic groups and geographical regions in India. In this study, we have investigated Kachhiya Patel, Dhodia Patel, Modh Bania, and Muslim communities of Surat, Gujarat to identify molecular defects causing ß-thalassemia in them. Covalent reverse dot blot hybridization technique was used to detect six common Indian ß-thalassemia mutations while the seventh mutation (619-bp deletion) was identified by PCR. The less common mutations were detected by amplification refractory mutation and the uncharacterized samples were directly sequenced. Characterization of ß-thalassemia mutations was carried out in a total of 175 unrelated ß-thalassemia trait cases. We identified IVS 1 nt 5 (G → C) in 31 out of 65 Muslims, codon (Cd) 41/42 (-CTTT) in 14 out of 16 in Modh Banias, Cd 15 (G → A) in 19 out of 24 Dhodia Patels. The most significant observation was an uncommon mutation; Cd 30 (G → A) detected in 61 out of 70 Kachhiya Patels. The 619-bp deletion was detected in 6 out of 10 Muslim-Memons. Many other rare mutations like Cd 15 (-T), Cd 8 (-AA), -88 (C → A), Capsite +1 (A → C), Cd 16(-C), and Cd 5 (-CT) were detected. To our knowledge, our study is the first to characterize ß-thalassemia mutations in the Kachhiya Patel community. This study will facilitate molecular analysis and prenatal diagnosis in these four communities.

Antenatal screening for identification of couples for prenatal diagnosis of severe hemoglobinopathies in surat, South gujarat.

PURPOSE: Our aim was to identify couples at risk of having a homozygous or compound heterozygous child with a severe hemoglobinopathy by antenatal screening and prenatal diagnosis in Surat, South Gujarat. METHOD: Pregnant women were screened for hemoglobinopathies by means of red cell indices, the solubility test, cellulose acetate electrophoresis tests, and confirmation by HPLC. Husbands of the pregnant women having hemoglobinopathies were counseled and screened for hemoglobinopathies. The couples at risk were again counseled and referred to the National Institute of Immunohematology, where mutations in parents and fetuses were identified by molecular analysis. After prenatal diagnosis, the continuing pregnancies were followed up and infants were tested at birth. RESULTS: Out of 3,009 women, 37.04, 52.6, and 10.3 % were in the first, second, and third trimester of pregnancy, respectively. Among those having hemoglobinopathies, 102 (3.38 %) had the ß-thalassemia trait, 46 (1.5 %) the Sickle cell trait, and 26 (0.86) had hemoglobin variants like Hb DPunjab, Hb E, Hb DIran, Hb QIndia, Hb JParis-I, and Hb OIndonesia. Out of the 14 couples at risk of having an affected child, 11 (78.5 %) couples opted for prenatal diagnosis. Three fetuses had homozygous ß-thalassemia and hence the pregnancies were terminated. Follow up of normal or heterozygous fetuses confirmed the diagnosis. CONCLUSION: During antenatal screening, we found many Hb variants of ß and α globin chains. Late antenatal registration, non-cooperation of the husband for investigation, and refusal for prenatal diagnosis are the main hurdles in the hemoglobinopathy prevention program and awareness is necessary.

Impact of donor arm cleaning with different aseptic solutions for prevention of contamination in blood bags.

Transfusion associated sepsis cases are encountered occasionally and bacterial transmission remains the major cause. The goal of our study was to compare the efficacy of disinfectants in phlebotomy site preparation. After selection of donor the antecubital fossa area of the arm was disinfected with different types of disinfectants namely sprit (70% isopropyl alcohol), povidone iodine (0.5% w/v available iodine in distilled water), savlon (1.5% v/v chlorhexidine gluconate solution and 3.0% cetrimide solution) and combination of sprit and povidone iodine. Swabs were collected from 20 donors using a sterile forceps, after cleaning with different antiseptic solutions. Swab was streaked on blood agar plate aseptically and the plate was incubated at 37°C for 24 h. Colonies were counted and a single colony was re-cultured by growing on nutrient and Mac-Conkey agar. The biochemical characteristics were determined by performing Gram staining, Motility, Catalase and Oxidase tests. The mean values of colonies were significantly higher with savlon compared to other three solutions. The difference was statistically significant by "t" test (t values 1.7-3.0; P < 0.05). Staphylococcus epidermidis, Staphylococcus sp., Streptococcus sp., Micrococcus sp., Bacillus megaterium and Bacillus cereus were the organisms identified. After completion of bleeding, samples from the bag were aseptically inoculated in aerobic and anaerobic culture bottles to be tested on BacT/Alert system. The bag containing donor's blood did not show any contamination when three cleanings were carried out using sprit, povidone iodine and spirit respectively.

Prevalence and hematological profile of ß-thalassemia and sickle cell anemia in four communities of Surat city.

BACKGROUND: From the data of transfusion-dependent thalassemia major cases, the 4 communities (Muslim, Dhodia Patel, Kachhiya Patel, and Modh Bania) with high prevalence but not studied methodically were selected. AIM: The aim of this study is to find prevalence of ß-thalassemia and sickle cell anemia in 4 selected communities and also to evaluate hematological profile in them. MATERIALS AND METHODS: For screening of ß-thalassemia trait (BTT) and sickle cell trait (SCT), all samples were tested for red cell indices, solubility, HbA(2) level and doubtful cases confirmed on HPLC. STATISTICAL ANALYSIS: Mean ± SD, χ(2) and 't' tests were used to evaluate the significance. RESULTS AND CONCLUSION: Among 4 selected communities, the highest prevalence of BTT was observed in Modh Bania (6.2%) and Kachhiya Patel (6.05%) and that of SCT in Dhodia Patel (14.0%). Significantly higher prevalence of BTT was observed in Memon (P < 0.0001) and of SCT in Khalifa 6.6% (P < 0.0001) compared to other Muslim sub castes. Anemia was more prevalent in BTT compared to non-BTT and non-SCT subjects. 80% of Dhodia Patel non-BTT and non-SCT subjects showed microcytic red cell morphology. Their Mean ± SD Hb concentration was 12.1 ± 1.73, hence iron deficiency cannot be a sole reason. This community needs α-thalassemia and iron studies.

Association of ABO groups in malaria infection of variable severity.

BACKGROUND & OBJECTIVES: Literature reports several studies on ABO groups and malaria but a study with an adequate sample size and controls is not available. ABO groups are genetically controlled, hence, large sample size and comparison with population frequency is essential. To determine whether malaria infection with variable severity has correlation with ABO groups. STUDY DESIGN & METHODS: Blood samples of non-transfused malaria cases were obtained from pathology laboratories and transfused malaria patients' from Blood Bank. The malaria parasites were identified by examination of thick and thin smears. Control (normal population) included 11,303 students. RESULTS: The ABO group frequency of normal population was 'O' 32.3%, 'A' 22.2%, 'B' 36.7% and 'AB' 8.8%. The overall ABO group distribution in 8028 malaria cases was 'O' 30%, 'A' 24.6%, 'B' 35.5% and 'AB' 8.9%. 'A' group incidence was significantly higher than normal ('A' vs non-'A' = 15, df=1, p <0.001). ABO group frequencies were comparable within Plasmodium falciparum and P. vivax malaria. There was no significant difference in ABO group distribution in malaria patients having severe anemia or among transfused and nontransfused malaria cases. About 32% of P. falciparum cerebral malaria cases and 36% DIC cases were of 'A' group. Compared to 22.2% 'A' group in the population, malaria cases showed preponderance of 'A' group. Because of the small numbers statistical evaluation was not done. CONCLUSION: 'A' blood group is more susceptible to have malaria infection and risk of cerebral malaria and DIC in malaria is also more in 'A' group individuals.

Hemoglobinopathies in South Gujarat population and incidence of anemia in them.

OBJECTIVE: To Screen of South Gujarat population for determination of prevalence of different hemoglobinopathies particularly beta thalassemia trait (BTT) and sickle cell trait (SCT) and find out the incidence of anemia in them. MATERIAL AND METHODS: The present study screened 32,857 samples of students from different school and colleges in South Gujarat. Blood samples were initially tested for solubility test and complete hemogram on hematology analyzer. Samples having MCV (≤78), MCH (≤28) and/or positive solubility test were investigated for Hb electrophoresis on cellulose acetate membrane (pH 8.6). Hb A2 level ≥3.5% was considered as diagnostic for BTT. High performance liquid chromatography on Biorad Hb variant system was done on samples having doubtful results. RESULT: Overall prevalence of BTT and SCT in South Gujarat was 4.4% and 1.3% respectively. Gamit, Vasava, Chaudhary, and Mahyavanshi castes had high prevalence of BTT (15.9%, 13.6%, 12.6%, and 6.9%) as well as SCT (22.2%, 15.2, 22.3, and 4.2%) respectively. Other communities like Lohana (10.8%), Sindhi (10.2%), Prajapati (6.3%), and Ghanchi (6.2%) also showed higher prevalence of BTT. Incidence of mild to moderate anemia was higher in BTT and SCT compared to non-BTT or non-SCT subjects. CONCLUSION: Study suggests that BTT is the most prevalent hemoglobinopathy in South Gujarat. ß-thalassemia and Sickle cell anemia are highly prevalent in Mahyavanshi, Chaudhary, Gamit, Vasava and Rohit. Prajapati, Lohana, Leva Patel, and Ghanchi have ß- thalassemia risk. SCT is more frequently detected in Dhodia Patel and Kukanas.

Iron status of regular voluntary blood donors.

BACKGROUND: Our blood bank is a regional blood transfusion centre, which accepts blood only from voluntary donors. AIM: The aim is to study iron status of regular voluntary donors who donated their blood at least twice in a year. MATERIALS AND METHODS: Prior to blood donation, blood samples of 220 male and 30 female voluntary donors were collected. Control included 100 each male and female healthy individuals in the 18- to 60-year age group, who never donated blood and did not have any chronic infection. In the study and control groups, about 10% subjects consumed non-vegetarian diet. After investigation, 85 males and 56 females having haemoglobin (Hb) levels above 12.5 g/dl were selected as controls. Donors were divided into 50 blood donation categories. Majority of the donors in >50 donation category donated blood four times in a year, whereas the remaining donors donated two to three times per year. Haematological parameters were measured on fully automatic haematology analyzer, serum iron and total iron-binding capacity (TIBC) by biochemical methods, ferritin using ELISA kits and transferrin using immunoturbidometry kits. Iron/TIBC ratio x 100 gave percentage of transferrin saturation value. STATISTICAL ANALYSIS: Statistical evaluation was done by mean, standard deviation, pair t-test, chi(2) and anova (F-test). RESULTS: Preliminary analysis revealed that there was no significant difference in the iron profile of vegetarian and non-vegetarian subjects or controls and the donors donating <20 times. Significant increase or decrease was observed in mean values of various haematological and iron parameters in donors who donated blood for >20 times (P < 0.001), compared to controls. Anaemia, iron deficiency and depletion of iron stores were more prevalent in female donors (P < 0.05) compared to males and especially in those male donors who donated their blood for more than 20 times. CONCLUSION: Regular voluntary blood donors should receive iron supplementation to prevent iron deficiency and depletion in iron stores.

Potential of commercial anti-D reagents in the identification of partial D variants in Indian population.

BACKGROUND & OBJECTIVE: As partial D variants are of clinical importance in transfusion medicine, the present study aims to determine the efficiency of commercial anti-D reagents to identify partial D variants. METHODS: Forty two samples of known partial D identified in the Indian population were tested with seven commercial monoclonal anti-D reagents. RESULTS: Most of the monoclonal anti-D reagents gave strong positive reactions (24 to 59%) to weak positive (28 to 47%) with partial D cells. Polyclonal anti-D detected all partial D variants as RhD positive, though reacting weakly with the majority (83%) of them. All the seven commercial monoclonal anti-D reagents detected some variants as D negative. Analysis of pairs of these reagents showed that the combinations of reagents 1 & 2 and 1 & 6 could detect all partial D variants as RhD positive and hence can be used for donor testing. INTERPRETATION & CONCLUSION: Findings of our study showed that none of the monoclonal reagents when used individually could detect all partial D variants. A combination of two suitable anti-D reagents are necessary to detect maximum number of partial D variants.

Evaluation of single unit red cell transfusions given to adults during surgery.

BACKGROUND AND OBJECTIVE: The ready availability of blood in Surat city has resulted in its liberal use. Surat Raktadan Kendra and Research Centre (SRKRC) is 30 years old blood bank having license to supply blood components. In spite of publishing quarterly bulletin and organizing seminars to update clinicians' knowledge still we have not achieved rational use of blood. Present study has evaluated the use of single unit as well as whole blood transfusion for adult surgery cases. MATERIALS AND METHODS: A total of 8470 surgery cases in the age group of 19 years to 95 years receiving 13070 units of whole blood and 3761 units of red cell concentrate (RCC) during surgery were analyzed on the basis of incidence of single unit and whole blood transfusions, sex and Hb values using Microsoft Excel. RESULTS: Analysis revealed that 38.9% cases received single unit and 77.7% whole blood transfusions. Females more frequently received single unit transfusion than males (p< 0.001). The highest incidence of single unit (62.2%) and whole blood transfusion (95.2%) was observed for urogenital surgeries. The lowest incidence (22.9%) of single unit transfusion was for cancer surgeries. There was no significant difference in the mean Hb level at which male and female cases, received single unit transfusion (p>0.5). Mean Hb concentration was about 10.0 g/dl for patients receiving single unit of whole blood and 8 g/dl for those who received single unit of RCC. CONCLUSION: Surgery patients are receiving inappropriate transfusions. Continuous awareness programmes are needed to achieve judicious use of blood.

G6PD deficiency in Vataliya Prajapati community settled in Surat.

BACKGROUND: A Study on Vataliya Prajapati was published earlier but heterozygous females were not identified. AIMS: To compare incidence of glucose-6-phosphate dehydrogenase (G6PD) deficiency in random and unrelated subjects, present and previous study and as per their original habitat. Incidence of heterozygous deficiency and clinical implication of deficiency was also determined. SETTINGS AND DESIGN: Camps were organized in Katargaon and Amroli regions. Blood specimens, with relevant demographic information, were collected from those who attended the camp. METHODS AND MATERIAL: A total of 1644 random blood samples were collected from 404 families participating in the camps. Nitroblue tetrazolium dye test was used for G6PD deficiency screening and quantitative assay for measurement of G6PD enzyme activity. STATISTICAL ANALYSIS USED: Chi2 test was used to evaluate significance and mean values were compared by the Student's ''t'' test. RESULTS: Incidence of G6PD deficiency was found as 22% among all the random samples tested. However, the G6PD deficiency among unrelated members was 27.9% in males and 12.4% (P< 0.001,df 1). The 13.9% of the females with heterozygous G6PD deficient status, together with the homozygous deficient phenotype makes the incidence comparable with males. Incidence of deficiency was comparable with previous study, in Katargam and Amroli and in Amerli and Bhavganar districts. Deficient subjects had mild anemia and hemolytic crisis rarely occurred. CONCLUSION: Vataliya Prajapatis have high incidence of G6PD deficiency without severe chronic hemolytic anemia. However before prescribing medicines physician should know the G6PD status of a Vataliya Prajapati patient.

Hematological findings and severity of G6PD deficiency in Vataliya Prajapati subjects.

OBJECTIVES: The present study was undertaken to assess the clinical implication of G6PD deficiency in Vataliya Prajapati (VP) subjects in Surat. METHOD: Blood samples of 954 children and 690 adults were collected in camps. Cord blood samples of 57 neonates born to VP mother were also collected. Medical history and other relevant information of all subjects were obtained. Samples were screened for G6PD deficiency by NBT test and the enzyme activity was estimated by WHO method. Hematological parameters were measured on hematology analyzer while reticulocyte count was measured using new methylene blue dye. RESULTS: The G6PD enzyme deficiency was detected in 27.5% males and 12.8% females. The enzyme levels in deficient subjects suggested class II variant. Hematological studies indicated mild anemia in G6PD deficient persons. Reticulocyte count was slightly raised (p <0.05). Out of eight G6PD deficient neonates one developed mild jaundice. Five deficient male adults gave the history of hemolytic crisis, three of them had typhoid, one tuberculosis and remaining one had fever of unknown origin. CONCLUSION: G6PD deficiency in majority of Vataliya Prajapati subjects is of mild type. However it is essential to test every Vataliya Prajapati subject for G6PD deficiency as certain infections and drugs can cause crisis in deficient person.