Comparison of estimation of volume of fetomaternal hemorrhage using Kleihauer-Betke test and microcolumn gel method in D-negative nonisoimmunized mothers.

In this study we assessed the efficay of the microcolumn gel method in the detection and quantification of the volume of fetomaternal hemorrhage (FMH) in comparison with the Kleihauer-Betke test (KB) in nonisoimmunized D- mothers. We collected blood samples from 80 D- indirect antiglobulin test-negative mothers over a span of more than 1 year. FMH was determined by KB and microcolumn gel method, and the results were compared. FMH was recorded as less than 4 mL by KB if no fetal cells were seen after examining 25 fields using 10x objective. If fetal cells were seen, slides were examined furhter to quantify FMH. By microcolumn gel method, FMH was reported as less than 0.1 percent, 0.1 percent, 0.2 percent, and 0.4 percent or greater. None of the patients had FMH greater than 15 mL by KB . Sixty-two patients (77.5%) had FMH less than 4mL by KB. In all these cases , FMH was less than or equal 0.2 percent (approximately 4mL) by microcolumn gel method. The mean volume of FMH in the remaining 18 (22.5%) cases by KB was 8.3 ± 1.7 mL. Fifteen (83.3%) of these 18 cases had FMH of at least 0.4 percent (approximately 8 mL) by gel technology. Three cases (16.7%) that differed from KB results had FMH of 0.2 percent by microcolumn gel method with a maximal FMH of 6.4 mL by KB. FMH was significanlty increased in cesarean delivery (mean FMH 9.5 ± 0.8 mL, range 7.9-10.4 mL, p=0.001) abd abtepartum hemorrahge (mean FMH 9.5 ± 0.9 mL, range 7.9-10.4 mL, p< 0.001). Microcolumn gel method is an effective screening test . Technologies like KB and flow cytometry are better options for detecting a large volume of FMh. Antepartum hemorrhage and cesarean delivery are risk factors for FMH. the 300-µg dose of cases. We need to analyze the relative cost-effectiveness of universal administration of 300µg of Rh immune globulin vs. FMH quantitation with subsequent administration of titrated doses.

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.

Algorithm for recall of HIV reactive Indian blood donors by sequential immunoassays enables selective donor referral for counseling.

BACKGROUND: HIV/AIDS pandemic brought into focus the importance of safe blood donor pool. AIMS: To analyze true seroprevalence of HIV infection in our blood donors and devise an algorithm for donor recall avoiding unnecessary referrals to voluntary counseling and testing centre (VCTC). MATERIALS AND METHODS: 39,784 blood units were screened for anti-HIV 1/2 using ELISA immunoassay (IA-1). Samples which were repeat reactive on IA-1 were further tested using two different immunoassays (IA-2 and IA-3) and Western blot (WB). Based on results of these sequential IAs and WB, an algorithm for recall of true HIV seroreactive blood donors is suggested for countries like India where nucleic acid testing or p24 antigen assays are not mandatory and given the limited resources may not be feasible. RESULTS: The anti-HIV seroreactivity by repeat IA-1, IA-2, IA-3 and WB were 0.16%, 0.11%, 0.098% and 0.07% respectively. Of the 44 IA-1 reactive samples, 95.2% (20/21) of the seroreactive samples by both IA-2 and IA-3 were also WB positive and 100% (6/6) of the non-reactive samples by these IAs were WB negative. IA signal/cutoff ratio was significantly low in biological false reactive donors. WB indeterminate results were largely due to non-specific reactivity to gag protein (p55). CONCLUSIONS: HIV seroreactivity by sequential immunoassays (IA-1, IA-2 and IA-3; comparable to WHO Strategy-III) prior to donor recall results in decreased referral to VCTC as compared to single IA (WHO Strategy-I) being followed currently in India. Moreover, this strategy will repose donor confidence in our blood transfusion services and strengthen voluntary blood donation program.

Change in tissue thromboplastin content of brain following trauma.

BACKGROUND: Tissue thromboplastin (TTP) is an integral membrane protein contributing to coagulopathy after trauma of brain, which is a rich source of TTP. AIMS: A study was undertaken to establish the TTP content of various areas of normal brain and estimate the changes in TTP activity of brain in response to varying degrees of trauma. MATERIALS AND METHODS: Samples from different areas of brain of ten cadavers were used as controls and they were compared with contused brain tissue obtained after surgery in 25 head injury (HI) patients of varying severity. RESULTS: In the study group, the TTP activity of the frontal, parietal, and temporal lobes after HI was significantly raised in contrast to that of the control group. The TTP activity was also significantly higher in the severe HI patients than those having moderate HI. The mode of injury and the time lapse after HI had no significant bearing on the TTP activity. Subjects above 40 years of age demonstrated a higher mean TTP activity after HI, though it was not statistically significant. CONCLUSION: The study provides quantitative data on TTP activity of normal brain and highlights the role of TTP in coagulopathy following HI through its increased activity after HI, more so in the severe HI group.

Alternative splicing of Drosophila calcium/calmodulin-dependent protein kinase II regulates substrate specificity and activation.

Drosophila calcium/calmodulin-dependent protein kinase II is alternatively spliced to generate multiple isoforms that vary only in a region between the calmodulin-binding domain and the association domain. This variation has been shown to modulate activation of the enzyme by calmodulin. In this study we examine the ability of seven of the Drosophila isoforms to phosphorylate purified protein substrates and to be inhibited by a substrate analog, and the response of six of the isoforms to a mutant form of calmodulin (V91G) that was isolated in a genetic screen. Significant variation in Kms for Eag, a potassium channel, and Adf-1, a transcription factor, were found. In the case of the a peptide inhibitor, AC3I, there were significant variations in Ki between isoforms. Kact for V91G calmodulin was increased for all of the isoforms. In addition, one isoform, RI, exhibited a lower Vmax when assayed with this mutant CaM. These results indicate that the variable domain of calcium/calmodulin-dependent protein kinase II is capable of altering the substrate specificity of the catalytic domain and the activation response to calmodulin.

Dysmyelopoietic Features and Bone Marrow Histology in 30 Cases of Primary Myelodysplastic Syndromes.

Thirty cases of primary myelodysplastic syndromes (MDS) were diagnosed according to the FAB criteria. The age ranged between 5-78 years, the median age being 38 years. Twelve (40%) patients were younger than 30 years of age. Male female ratio was 2:1. Nine cases each were diagnosed as refractory anaemia (RA) and refractory anaemia with excess of blasts (RAEB) whereas 12 were diagnosed as RAEB in transformation (RAEB-T). Significantly more RA and RAEB-T cases showed dyserythropoietic features when compared to RAEB. The frequency of dysgranulopoiesis and dysmegakaryopoiesis was similar in all 3 FAB subgroups. Abnormal localisation of immature precursors (ALIP) was present in only 36.6% of the patients and was seen in 58.3% of RAEB-T. When present it appeared to indicate a worse prognosis. Six of the 12 cases showing this finding progressed either to acute myeloblastic leukemia or from RA to RAEB or RAEB-T.

Functional and ultrastructural changes of platelets in malarial infection.

This paper describes changes in the circulating platelets of 25 patients with acute malaria within 2 to 6 days of onset of illness. Thrombocytopenia was observed in 10 out of 15 patients with Plasmodium falciparum infection, and in 4 out of 9 patients with P. vivax infection. One patient with a mixed infection of both species had a disseminated intravascular coagulation. Platelet antibody was detected in the sera of 8 out of 11 cases by the complement lysis inhibition technique and indirect immunofluorescence. The mean platelet antibody concentrations in the sera of 11 patients and 53 control subjects were 122.70 +/- 80.25 ng/10(7) platelets and 36.69 +/- 18.72 ng/10(7) platelets, respectively. An inverse relationship between the platelet count and platelet antibody levels in serum supported the view that thrombocytopenia in malaria may be partly immune-mediated. Platelet aggregation responses to agonists such as ADP, adrenaline, collagen and ristocetin revealed hyperactivity. Ultrastructural study of unstimulated platelets from patients revealed several changes such as centralization of dense granules, glycogen depletion, and formation of pseudopods and microaggregates, indicating in vivo activation of the platelets, which may also lead to thrombocytopenia.

Enteral nutrition in surgical patients.

BACKGROUND: Malnutrition is common in patients admitted for surgery and is a major cause of increased morbidity and mortality. Nutritional support has been shown to be of help in reducing complications. Parenteral nutrition and commercially available enteral diets are expensive, so the efficacy of a 'home-brew' enteral diet was studied in such patients. METHODS: Forty malnourished patients, 20 with benign disease and 20 with malignancy, were administered a 'home-brew' enteral diet (1140 calories and 60 g protein per litre) perioperatively for 14 days. They received 2500 to 4000 calories per day according to their requirement. Weight, triceps skinfold thickness, midarm circumference, serum albumin and transferrin, absolute lymphocyte count and creatinine-height index were monitored on days 0, 7 and 14. Nitrogen balance was estimated on alternate days and the results of the two groups were compared. RESULTS: Weight, skinfold thickness and midarm circumference did not change. Serum albumin levels showed a rise in the benign group by day 7, but the rise attained significance in the malignancy group by day 14. A similar pattern was observed in transferrin levels and there was a significant correlation (r = 0.652, p < 0.001) between albumin and transferrin levels. A positive nitrogen balance was attained earlier in the benign group (4.3 v. 5.8 days, p < 0.001). The creatinine-height index showed a rise in both groups by day 7 and a further rise by day 14. Diarrhoea was the commonest complication but was easily controlled with loperamide. The efficacy of the diet was evidenced by the improvement recorded in various objective nutritional parameters and complications were minimal. CONCLUSION: This cost-effective diet may be used for nutritional support with good results in patients in India and other developing countries.

Hemostatic alterations in non-cirrhotic portal fibrosis, extrahepatic portal venous obstruction and Budd-Chiari syndrome.

We studied the coagulation function in ten patients each with non-cirrhotic portal fibrosis (NCPF), extrahepatic portal venous obstruction (EHPO) and Budd-Chiari syndrome (BCS), conditions where venous thrombosis in the hepatic vasculature is a common denominator. Prothrombin time, partial thromboplastin time with kaolin (PTTK) and thrombin time were normal in patients with NCPF and EHPO. However, in BCS the PTTK was prolonged, with a mean test/control ratio of 1.68 +/- 0.11. Fibrin degradation products were absent in all patients. Platelet aggregation tests showed hypoaggregability in all patients with NCPF. They were normal in patients with EHPO. However, two of ten BCS patients showed hyperaggregability, coinciding with a recent onset of illness in one patient. In conclusion, coagulation abnormalities appear unlikely to be the cause of thrombosis in patients with NCPF and EHPO. Further studies are required to substantiate the findings of hyperaggregability of platelets in BCS.

Coagulation disorders and neo-membrane formation in the pathogenesis of chronic subdural haematoma.

Haematological investigations in the haematoma fluid and venous blood along with histological changes in the haematoma membrane were studied in twenty five patients of chronic subdural haematoma. Their median age was 60 years with a marked male predominance. Haemoglobin level in the haematoma fluid was unrelated to its age. Prolongation of prothrombin time and activated partial thromboplastin time was observed in all the haematoma fluid samples. Fibrinogen and antithrombin III levels were significantly decreased in haematoma fluid and fibrin degradation products were positive in 44 of these specimens. The neo-membrane showed 'giant' capillaries and abundant eosinophils with an inverse relationship between the presence of inflammatory cells andfibroblasts. These findings suggest a localized consumption coagulopathy and hyperfibrinolysis with reactive formation of a leaky neo-membrane resulting in recurrent haemorrhages and progressive enlargement of chronic subdural haematoma.

Fibrinolytic activity in human malignant tumors.

We have studied fibrinolytic activity of 12 cases of infiltrating duct carcinoma of breast (7 metastatic and 5 non-metastatic) and ten cases of adenocarcinoma of gastrointestinal tract (5 each of metastatic and nonmetastatic), and compared with some of their normal tissue counterparts. Both metastatic and non-metastatic tumors of breast and gastrointestinal tract had significantly higher levels of fibrinolytic activity as compared to normal tissues. Though mean fibrinolytic activity (expressed as ug/ml of urokinase activity) of metastatic tumors of breast and gut had higher values as compared to non-metastatic counterparts, however it did not reach statistical significance.

Histological and clinical evolution patterns of chronic myelocytic leukemia.

Trephine biopsies of 101 chronic myelocytic leukaemia (CML) patients were analysed to study the relationship between initial and subsequent histological features vis-a-vis clinical behaviour of the disease. The patients with blast crisis at presentation were excluded. At diagnosis 62 (61.4%) patients revealed granulocytic-megakaryocytic (gran-meg) proliferation whereas granulocytic (gran) proliferation was found in 39 (38.6%) patients. Gran pattern at diagnosis was associated with shorter survival and early evolution into blast crisis (36.8%) in 12 months, although the difference in the total incidence of blast crisis between the two histological groups was not statistically significant. Myelofibrosis was detected in more number of cases on follow up (89.1%) as compared to the initial biopsies (80.2%). However myelofibrosis did not correlate with initial cellular composition, overall survival or the phase of CML (P > 0.05). Transition from one histological type to another was observed in 15 out of 60 (25%) cases while remaining in the chronic phase.

Aplastic anaemia versus hypocellular myelodysplastic syndrome.

It is important to differentiate non-dyplastic aplastic anaemia from hypocellular myelodysplastic syndrome (MDS). Four patients presenting with hypocellular bone marrow and different evolution patterns are being described. Certain morphological features and variable hypocellularity were found to be useful indices for this purpose.

Pure red cell aplasia associated with essential thrombocythemia (a case report).

A rare case of erythroblastopenia associated with essential thrombocythemia (ET) is described. The patient had markedly elevated platelet count (5200 x 10(9)/1) and significant platelet dysfunction leading to extensive soft tissue and gastrointestinal hemorrhage. There was paucity of erythroid precursors in the bone marrow - a feature hitherto undescribed in ET. The thrombocytosis responded to well busulphan therapy but patient succumbed to fulminant infection consequent upon drug induced neutropenia.