Endothelial nitric oxide synthase (eNOS) is generally expressed in endocardial cells, vascular endothelial cells and ventricular myocytes. However, there is no experimental study elucidating the relationship between cardiac eNOS expression and elevated plasma viscosity in low oxygen delivery pathological conditions such as hemorrhagic shock-resuscitation and hemodilution. This study tested the hypothesis that elevated plasma viscosity increases cardiac eNOS expression in a hemodilution model, leading to positive effects on cardiac performance.
Materials and Methods:
Two groups of golden Syrian hamster underwent an acute isovolemic hemodilution where 40% of blood volume was exchanged with 2% (low-viscogenic plasma expander [LVPE]) or 6% (high-viscogenic plasma expander [HVPE]) of dextran 2000 kDa. In control group, experiment was performed without hemodilution. All groups were performed in awake condition. Experimental parameters, i.e., mean arterial blood pressure (MAP), heart rate, hematocrit, blood gas content and viscosity, were measured. The eNOS expression was evaluated by eNOS Western blot analysis.
After hemodilution, MAP decreased to 72% and 93% of baseline in the LVPE and HVPE, respectively. Furthermore, pO2 in the LVPE group increased highest among the groups. Plasma viscosity in the HVPE group was significantly higher than that in control and LVPE groups. The expression of eNOS in the HVPE group showed higher intensity compared to other groups, especially compared with the control group.
Our results demonstrated that cardiac eNOS has responded to plasma viscosity modulation with HVPE and LVPE. This particularly supports the previous studies that revealed the positive effects on cardiac function in animals hemodiluted with HVPE.
Cardiac endothelial nitric oxide synthase; hemodilution; plasma expander; plasma viscosity
Background and Aim:
Autoimmune hemolytic anemia (AIHA) is characterized by the production of autoantibodies directed against red cell antigens. Most patients of AIHA arrive in the emergency or out-patient department (OPD) with severe anemia requiring urgent blood transfusion. Here we share our experience of managing these patients with incompatible blood transfusions and suggest the minimal test required to assure patient safety.
Materials and Methods:
A total of 14 patients admitted with severe anemia, diagnosed with AIHA and requiring blood transfusion urgently were included in the study. A series of immunohematological investigations were performed to confirm the diagnosis and issue best match packed red blood cells (PRBC) to these patients.
A total of 167 PRBC units were crossmatched for 14 patients of which 46 units (28%) were found to be best match ones and 26 (56.5%) of these units were transfused. A mean turn around time of 222 min was observed in issuing the “best match” blood. Severe hemolysis was observed in all patients with a median hemoglobin increment of 0.88 g/dl after each unit PRBC transfusion.
Decision to transfuse in AIHA should be based on the clinical condition of the patient. No critical patient should be denied blood transfusion due to serological incompatibility. Minimum investigations such as direct antiglobulin test (DAT), antibody screening and autocontrol should be performed to ensure transfusion safety in patients. All transfusion services should be capable of issuing “best match” PRBCs in AIHA.
Alloantibody; autoantibody; autoimmune hemolytic anemia; best match blood; gel technology
The aim of the present study was to determine the prevalence of syphilis infection by Treponema pallidum hemagglutination assay (TPHA) among blood donors in Delhi and to study their correlation with other markers of transfusion transmitted infections such as hepatitis C virus (HCV), human immunodeficiency virus (HIV) and hepatitis B surface antigen (HBsAg) so as to establish the utility of TPHA over and above venereal diseases research laboratory test (VDRL), not only as a marker for testing T. pallidum infection, but also as a marker of high risk behavior.
Materials and Methods:
This prospective study was carried out in the Regional Blood Transfusion Centre, Lady Hardinge Medical College and associated Sucheta Kriplani Hospital, New Delhi for a period of 2 years. Donated blood was screened for TPHA seroreactivity along with screening for anti HIV I and II, anti-HCV, HBsAg by third generation enzyme-linked immunosorbent assay test. A total of 8082 serum samples of blood donors were collected from healthy blood donors in our blood bank. They were classified into two groups- test group and control group based on TPHA positivity. The co-occurrence of HBsAg, HIV and HCV infection were determined in TPHA positive blood donors (test group) in comparison with TPHA negative blood donors (control group).
We found the TPHA seroreactivity to be 4.4% in Delhi's blood donors. Nearly 8.2% (663/8082) of the donated blood had serological evidence of infection by at least one pathogen (syphilis/HIV/hepatitis B virus/HCV) and 6.63% (44/663) donors with positive serology had multiple infections (two or more). Quadruple infection was seen in one donor, triple infection was seen in three donors and double infection was seen in 40 donors. Prevalence of HIV seroreactivity was found to be statistically significant and HCV seroreactivity statistically insignificant in TPHA positive group in comparison to TPHA negative group.
In our study, the TPHA seropositivity correlated with higher HIV and HCV seropositivity and the same correlation has been observed by several other studies also. In view of these observations, we propose that testing for syphilis by more sensitive and specific treponemal markers like TPHA rather than VDRL, rapid plasma reagin tests; as TPHA also has the added advantage of picking up all the high risk donors, whereas, VDRL picks up only currently infected donors. Moreover, TPHA should be continued as a marker of high risk behavior especially in high prevalence areas like India where we don’t have universal access to markers like nucleic acid amplification technique.
Syphilis; Treponema pallidum hemagglutination assay; transfusion transmitted infections; Co-infection
Maternal red blood cell (RBC) alloimmunization may lead to production of harmful antibodies that result in hemolytic disease of fetus and newborn (HDFN). There is insufficient data on the prevalence of HDFN due to RBC alloantibodies in the Malay neonatal population.
The aim of this study was to determine the incidence of HDFN in the Malay neonatal population due to clinically significant RBC alloantibodies.
Subjects and Methods:
A cross sectional study was conducted in Transfusion Medicine Unit, Hospital Universitiy Sains Malaysia over one year period from January to December 2009. A total of 5163 Malay pregnant women who attended labor room for delivery were collected and analyzed prospectively. The blood samples were subjected to the standard immunohematological procedure for RBC antibody screening and identification using reagents of Diamed-ID Gel microtyping system. All the newborns with RBC alloantibody were investigated for the evidence of HDFN.
Thirty (0.58%) women were found to have clinically significant RBC alloantibodies. Most of the alloantibodies belonged to Rhesus (Rh) system (56.7%) where anti-E (33.3%) was the most common followed by anti-D (10.0%). Rh antibodies were the main cause of HDFN in fourteen (0.27%) neonates. Anti-D and anti-c were identified to cause moderate to very severe HDFN.
With the low prevalence of clinically significant RBC alloantibodies and HDFN, routine antenatal antibody screening practice may not be advised as a routine practice at present, preferably reserved for those women of RhD negative or with history of HDFN, significantly of those attributed to anti-c.
Clinically significant alloantibodies; HDFN; Malay
The knowledge of physicians concerning blood transfusion has a significant impact on the optimal use of blood and blood products. The aim was to survey their knowledge regarding this area and identify whether additional training is required.
Material and Methods:
This cross-sectional study was conducted on all 1st year resident physicians at Shiraz University of Medical Sciences, Iran in 2011. The questionnaire solicited information on demographic variables, knowledge regarding transfusion medicine, education and experience regarding blood transfusion.
The mean total knowledge score regarding transfusion medicine was 15.44 ± 3.3 (7-25) out of 29. Only about one-fourth (27.4%) replied correctly to over 60% of questions. The mean score of knowledge was higher among residents who stated that they received special training regarding blood transfusion in their medical courses (P < 0.01). Seventy-five percent of residents believed that they had received insufficient education and 97.8% believed that they need additional training.
The results reflect the uncertainties among resident physicians regarding blood transfusion. It has been suggested that a special transfusion medicine educational program should be added to the medical education curriculum.
Blood transfusion; education; knowledge; medical education; physician; transfusion medicine
Background and Objectives:
The study was undertaken with the objective to provide data on the ABO and Rh(D) blood group distribution and gene frequency across India.
Materials and Methods:
A total of 10,000 healthy blood donors donating in blood banks situated in five different geographical regions of the country (North, South, East and Center) were included in the study. ABO and Rh (D) grouping was performed on all these samples. Data on the frequency of ABO and Rh(D) blood groups was reported in simple numbers and percentages.
The study showed that O was the most common blood group (37.12%) in the country closely followed by B at 32.26%, followed by A at 22.88% while AB was the least prevalent group at 7.74%. 94.61% of the donor population was Rh positive and the rest were Rh negative. Regional variations were observed in the distribution. Using the maximum likelihood method, the frequencies of the IA, IB and IO alleles were calculated and tested according to the Hardy Weinberg law of Equilibrium. The calculated gene frequencies are 0.1653 for IA (p), 0.2254 for IB (q) and 0.6093 for IO (r). In Indian Population, O (r) records the highest value followed by B (q) and A (p); O > B > A.
The study provides information about the relative distribution of various alleles in the Indian population both on a pan-India basis as well as region-wise. This vital information may be helpful in planning for future health challenges, particularly planning with regards to blood transfusion services.
ABO; blood group; rhesus
Introduction & Aims:
Though platelet transfusions have greatly reduced the incidence of major haemorrhagic complications associated with the management of haematological and oncological disorders, refractoriness to infused platelets becomes a major clinical problem for many of these patients.
Materials and methods:
The present study was done to determine the percentage of platelet alloimmunisation due to platelet-reactive antibodies in 340 patients with hematologic or oncologic diseases who had received multiple transfusions (> 10) of blood and blood components and showed platelet refractoriness in 1-hour post transfusion sample.
Platelet-reactive antibodies were detected in the sera of 127 out of 340 patients (37.35%) who received multiple transfusions (> 10) and showed platelet refractoriness.
Platelet-reactive antibodies appear to be an important cause of platelet refractoriness in patients of acute leukaemia, aplastic anaemia, NHL, MDS and multiple myeloma receiving multiple platelet transfusions. Platelet refractoriness in patients of ITP and chronic leukaemia appears to be due to other causes and not due to platelet-reactive antibodies.
Platelet alloimmunization; platelet immunofluorescent test; refractoriness
Beta-thalassemia is one of the most prevalent autosomal disorders, which affect more than 400,000 newborn per year worldwide. In India, the carrier rate of beta-thalassemia varies from 3-17%. The overall rate of alloimmunization in thalassemia patients has been reported to be 5-30% in the world, which is mostly contributed by the alloimmunization to minor blood group antigen. Among Asians, the incidence of red cell alloimmunization is 22%. The recommended treatment for beta-thalassemia major is regular blood transfusion every 3 to 4 weeks. The development of anti-red cell antibodies (alloantibodies and/or autoantibodies) can significantly complicate transfusion therapy. Alloantibodies are commonly associated with red cell hemolysis. Red cell autoantibodies appear less frequently, but they can result in clinical hemolysis called autoimmune hemolytic anemia (AIHA), and in difficulty in cross-matching blood. Patients with autoantibodies may have a higher transfusion rate and often require immunosuppressive drugs or alternative treatments including intravenous immunoglobulin (IVIg) and rituximab (anti-CD20 monoclonal antibody).
Thalassemia major; auto immune hemolytic anemia; alloimmunization
Transfusion-related acute lung injury (TRALI) is a serious clinical syndrome associated with the transfusion of plasmacontaining blood components. Recently, TRALI has come to be recognized as the leading cause of transfusion-related mortality. This complication typically presents as shortness of breath, hypoxemia, hypotension, fever, and non cardiogenic pulmonary edema, occurring within 6 h after transfusion. Although the mechanism of TRALI has not been exactly known, it has been associated with human leukocyte antigen antibodies and with biologically active mediators in stored cellular blood components. We, hereby, present a case of a patient with dengue fever who developed acute lung injury (ALI), presumably TRALI, after transfusion of platelet concentrates. He was treated with supportive measures and mechanical ventilation. Greater knowledge and increased awareness especially amongst the clinicians regarding TRALI is needed for prevention and treatment of this potentially severe complication of blood/component transfusion.
Acute lung injury; blood transfusion; non cardiogenic pulmonary edema; transfusion-related acute lung injury
Donor notification and post-donation counseling is an essential role of blood bank. If a donor is reactive for any marker, the blood bank counselor, informs the donor and advices him/her to report to the blood bank for further counseling and management. The counselor at our blood bank informed a young female voluntary donor to be reactive for HIV both with ELISA as well as NAT. When the donor reported to blood bank, the repeat testing was negative and no history of high risk behavior could be elicited. The hospital information system (HIS) records were checked again immediately for clarification and showed consistency with her demographic profile. But when her manual records and donor questionnaire were retrieved, showed information displayed in the HIS system was wrongly interpreted by the counselor. In this era of information technology being highly advanced, the role of manual record keeping is still the gold standard.
Donor notification; hospital information system; reactive donor
A 66-year-old female patient presented with complaints of abdominal discomfort, pigmentation and numbness of both lower limbs for 3 years duration. On examination, she had erythema of the face and palms. Investigations revealed high hemoglobin (Hb), hematocrit (Hct) and erythropoietin. Ultrasonography abdomen showed large uterine fibroid. As there are increased tendencies of thromboembolic episodes in patients undergoing surgeries with such high Hb and Hct, a target to achieve a Hb of 15g/dl and Hct of 45 was set in the patient. Repeated phlebotomies were done over 10 days with oral hydration only and the Hb was brought down to 18 g/dl on the day prior to surgery. On the day of surgery, pre-operative phlebotomy was done so as to remove 2 units of 350 ml blood and was transfused intraoperatively to combat blood loss. Post-operatively Hb was 12.4 g %. Patient was discharged on the 10th post-operative day with further follow-up evaluations being uneventful.
Leiomyomatous erythrocytosis; perioperative hemodilution; phlebotomy
Plasma is used to correct coagulopathies, but not all coagulation abnormalities are clinically significant enough to require correction before an invasive procedure. We report an 82-year-old female who, in response to a mildly prolonged INR of unknown etiology, was unnecessarily transfused with plasma in advance of elective surgery. The patient suffered a moderately severe transfusion reaction, including hives and voice hoarseness, which caused a 4-week delay in her surgery. This delay and adverse reaction could have been avoided had the principles of evidence based plasma therapy, which we herein review, been followed and if the etiology of the mildly elevated INR been investigated before the day of her surgery.
Allergic reaction; guidelines; plasma; transfusion
Blood transfusion carries the risk of transmission of several infectious agents. The latest method for blood screening, nucleic acid testing is not affordable in developing countries.
The study was aimed to find response to post-donation counseling for reactive markers among the voluntary blood donors donating in blood donation camps.
Material and Methods:
This 1 year study was conducted in 2011. Transfusion transmitted infections testing was performed by routine enzyme linked immunosorbent assay method. The initial human immunodeficiency virus (HIV) reactive donors who returned back to the blood bank were confidentially counseled and referred to the Integrated Counseling Cum Testing Center (ICTC). The hepatitis B surface antigen (HBsAg) and anti-hepatitis C virus (HCV) reactive donors were referred to the gastroenterology department for confirmation by qualitative polymerase chain reaction (PCR, Roche Diagnostics, Germany) and followed-up.
Twenty seven thousand two hundred forty six 27,246 units were collected during the survey. One hundred twenty nine129 units were reactive for HIV 1 and 2, 99 were reactive for HCV, 206 for hepatitis B virus (HBV). Of these reactive donors, 138 could be personally communicated. Out of 47, 27 donors who returned for counseling were initially reactive for HIV 1 and 2, 8 for HBsAg and 12 for anti-HCV. Two were positive for HBV deoxyribonucleic acid and one was positive for HCV ribonucleic acid. The HIV positivity was detected in 1 of 27 donors at ICTC.
The response to the post-donation counseling appears in this study to be only 34% (47/138), which is still a challenge.
Integrated counseling and testing center; post-donation counseling; transfusion transmitted infections
The development of anti-red blood cell antibodies (both allo-and autoantibodies) remains a major problem in thalassemia major patients. We studied the frequency of red blood cell (RBC) alloimmunization and autoimmunization among thalassemia patients who received regular transfusions at our center and analyzed the factors, which may be responsible for development of these antibodies.
Materials and Methods:
The study was carried out on 319 multiply transfused patients with β-thalassemia major registered with thalassemia clinic at our institute. Clinical and transfusion records of all the patients were examined for age of patients, age at initiation of transfusion therapy, total number of blood units transfused, transfusion interval, status of splenectomy or other interventions. Alloantibody screening and identification was done using three cell and 11 cell panel (Diapanel, Bio-rad, Switzerland) respectively. To detect autoantibodies, autocontrol was carried out using polyspecific coombs (IgG + C3d) gel cards.
Eighteen patients out of total 319 patients (5.64%) developed alloantibodies and 90 (28.2%) developed autoantibodies. Nine out of 18 patients with alloantibodies also had autoantibodies. Age at first transfusion was significantly higher in alloimmunized than non-immunized patients (P = 0.042). Out of 23 alloantibodies, 52.17% belonged to Rh blood group system (Anti-E = 17%, Anti D = 13%, Anti-C = 13%, Anti-Cw = 9%), 35% belonged to Kell blood group system, 9% of Kidd and 4% of Xg blood group system.
Alloimmunization was detected in 5.64% of multitransfused thalassemia patients. Rh and Kell blood group system antibodies accounted for more than 80% of alloantibodies. This study re-emphasizes the need for RBC antigen typing before first transfusion and issue of antigen matched blood (at least for Rh and Kell antigen). Early institution of transfusion therapy after diagnosis is another means of decreasing alloimmunization.
Alloimmunization; autoimmunization; thalassemia major; transfusion
Random donor platelet (RDP) is not sufficient to improve the platelet count in most thrombocytopenic patients. Single donor platelet (SDP) or buffy coat pooled platelet (BCPP) are the two choices to provide a full therapeutic dose of platelets. However, there are constraints in the preparation of SDP due to stringent donor selection procedure, time required for procedure, and need of special expensive equipments and kits. BCPP is widely practiced, especially in the European countries, since 1995. In India, we decided to adopt the procedure of buffy coat pooling of platelets, especially for economically backward patients and for emergencies. This study was prospectively conducted from September 2009 to September 2010. A total of 129 units of BCPP [tested prior for viral markers by enzyme-linked immunosorbent assay (ELISA) and individual donor nucleic acid amplification test (ID-NAT)] were issued to 129 patients suffering from dengue and were included in this study. For comparison between efficacy of SDP and BCCP, patients were divided into two groups of 50 each. The post-transfusion platelet counts of the patients were noted after 2 hours of transfusion for each type of component. The platelet yield varied from 2.5 to 4.4 Χ 1011 in BCPP samples. The samples analyzed were sterile without any contamination. The different biochemical parameters were analyzed in detail. The observed post-transfusion platelet recovery and corrected count increment (CCI) at 1 hour and 24 hours after BCPP transfusion were similar to that after SDP transfusion. Hence, we concluded that BCPP can be a low cost alternative to SDP in the times of emergencies like dengue and non-affordability by the patient for SDP.
buffy coat; buffy coat pooling of platelets; random donor platelets; single donor platelets
A prospective study was undertaken to evaluate the use of 2% (w/v) alcoholic chlorhexidine gluconate (2% AlcCHG) in donor arm preparation, to monitor the contamination rate of blood products after the collection and to find incidence of transfusion associated bacteremia.
Settings and Design:
Optimal skin antisepsis of the phlebotomy site is essential to minimize the risk of contamination. Food and Drug Administration (FDA) in India has recommended antisepsis with three-step regimen of spirit-10% povidone iodine-spirit for donor arm antisepsis, but not with chlorhexidine, which is recommended by many other authors.
Material and Methods:
A total of 795 donors were studied from July 2011 to January 2012. Spirit-10% povidone iodine-spirit was used for 398 donors and 2% AlcCHG was used for 397 donors with the two-step method for arm antisepsis. Swabs were collected before and after use of antiseptic agents for all the donors. All the blood products collected from donors with growth in post-antisepsis swabs were cultured. A total of 123 various blood products were cultured irrespective of the method and result of antisepsis was observed. A total of seven patients had mild transfusion reaction. The transfused blood products, blood and urine specimen of the patients who had transfusion reaction were also cultured.
Seven donors out of 398 donors had growth in post-antisepsis swab with spirit-10% povidone iodine-spirit protocol and three donors out of 397 donors had growth in post-antisepsis swab with 2% AlcCHG protocol. All blood products collected from donors who had growth in post-antisepsis swabs when cultured had no growth. There was no contamination of blood products.
Two percent (w/v) alcoholic chlorhexidine gluconate with two-step protocol can be used as an antiseptic agent for donor arm preparation without considerable cost difference. It is at par with spirit 10% povidone iodine spirit protocol as suggested by FDA in India. There was no reported transfusion associated bacteremia in the study period.
Bacteremia; blood donor arm; chlorhexidine gluconate
The clinically significant antibodies are those active at 37°C and/or by the indirect antiglobulin test. Most of the published literature refers to antibodies of Lewis blood group system to be insignificant, whereas antibodies to M and N blood groups are associated with variable clinical significance.
The aim of this study is to find the frequency and clinical significance of antibodies to M, N and Lewis blood group systems.
Settings and Design:
The study was carried out retrospectively from January 2009 to December 2012.
Materials and Methods:
Antibody screening was performed by solid phase red cell adherence (SPRCA) technique using four cell screening panel on a fully automated platform GALILEO (Immucor Inc. USA). In case of a positive antibody screen, antibody identification was performed using SPRCA (GALILEO, Immucor Inc. USA).
A total of 49,077 red cell antibody screens were performed and a total of 427 identifications of red cell antibodies were carried out. A total of 304 specific antibodies were detected: 8.22% of antibodies were of anti-M specificity and 2.96% were of anti-N specificity. Majority (84%) of anti-M and 77.78% of anti-N were of Immunoglobulin G (IgG) class reacting at 37°C. 1.31% of the antibodies were directed against Lewis system antigens of which 0.65% were anti-Lea and 0.65% were anti-Leb. Half of the Lewis system antibodies, i.e., 1 each of anti-Lea and anti-Leb were of IgG class.
Our study highlights the importance of detecting the thermal amplitude of antibodies with variable clinical significance especially if both IgG and IgM types of antibodies are associated with it so as to establish their clinical significance.
Anti-M; anti-N; clinical significance; Lewis
In the last few decades through an awareness of transfusion transmitted infections (TTI), a majority of countries have mandated serology based blood screening assays for Human immunodeficiency virus (HIV), Hepatitis C virus (HCV), and Hepatitis B virus (HBV). However, despite improved serology assays, the transfusion transmission of HIV, HCV, and HBV continues, primarily due to release of serology negative units that are infectious because of the window period (WP) and occult HBV infections (OBI). Effective mode of nucleic acid technology (NAT) testing of the viruses can be used to minimize the risk of TTIs. This review compiles the examples of NAT testing failures for all three viruses; analyzes the causes for failure, and the suggestions from retrospective studies to minimize such failures. The results suggest the safest path to be individual donation testing (ID) format for highest sensitivity, and detection of multiple regions for rapidly mutating and recombining viruses. The role of blood screening in the context of the donation and transfusion practices in India, the donor population, and the epidemiology is also discussed. World wide, as the public awareness of TTIs increases, as the recipient rights for safe blood are legally upheld, as the possibility to manage diseases such as hepatitis through expensive and prolonged treatment becomes accessible, and the societal responsibility to shoulder the health costs as in the case for HIV becomes routine, there is much to gain by preventing infections than treating diseases.
Donor look back; individual donation nucleic acid technology testing; multi pool nucleic acid technology testing; recipient trace back; transfusion transmitted viral infections; window period and occult infections