Red cell alloimunization and autoantibodies in transfusion dependent thalassemia patients of Jammu region

Background:Thalassemia is one of the most common genetic disorder of hemoglobin synthesis in Jammu region. Although RBC transfusion is life saving for these patients, it may be associated with some complications like RBC alloimmunization. Thus, alloimmunization against red blood cell antigens increases the need for transfusion and can significantly complicate transfusion therapy. Therefore, screening for unexpected antibodies should be a part of all pretransfusion testing,with antibody identification in the event of a positive result. The aim of the study was to determine the frequency of alloimmunization and autoimmunization and the most common alloantibodies involved.Methods:This was a descriptive study involving a total of 146 thalassemic patients in the age range of 2-32 years receiving regular blood transfusions, registered at SMGS blood bank, Jammu. Antibodies screening, antibody identification, and cross matching was doneon all patient samples included in the study, during the period between November 2014 and October 2015.Results:At the start of the study, 8 patients who tested positive for alloantibodies 3 patients had more than one antibody subtype. Anti-E was the commonest antibody found in 4 (50%) patients. Similarly, at the end of study, antibody screening and then identification revealed presence of antibodies in 10 patients. Only 1 patient had more than one antibody subtype. Anti E was again the commonest antibody found in 5 (50%) patients. Conclusions:The most common alloantibodies identified were anti Rh system antibodies (anti-E and anti-D) followed by Kell antibodies. In order to reduce alloimmunization, a policy for performing extended red cell phenotyping of these patients is essential and at least antigen E and Kell negative blood should be provided for transfusion to these patients.

Study of Cholesterol Levels in Ascitic Fluid to Differentiate Malignant from Non-Malignant Effusion.

Collection of more than normal quantity fluid in a serous cavity is called effusion. Itis classified as per location i.e. pericardial, pleural and peritoneal. Fluid collection inabdominal cavity is called ascites. Effusions are of two types - (1) Transudate (2)Exudate. Transudate develops as a result of physiological disturbances of circulationusually a rise in venous pressure or decrease in oncotic pressure, while exudateforms as a result of increased capillary permeability which is usually due toinflammation. Transudates usually have low specific gravity due to low proteincontent whereas exudates have high specific gravity due to high protein content.Transudates are usually associated with cardiac-, hepatic- or renal-disease. Whileexudates are caused by inflammatory conditions like tuberculosis and malignanciesof pelvic and abdominal organs. Recently fibronectin and cholesterol levels ofperitoneal fluid have been found to differentiate between ascites of differentaetiologies. The estimation of fibronectin levels in fluid is sensitive but complicated.So, in this study we have taken the value of cholesterol for the aetiologic diagnosisof ascites as a simple procedure.METHODSThe present descriptive study of 100 cases of ascites was conducted over a period of12 months i.e. January. 2019 to January 2020. Various fluids were collected fromOPD and IPD of Medical and Surgery units.RESULTSIn the present study, 84% of the cases were transudates while exudates accountedfor 16% of cases. Cirrhosis (65%) was the commonest cause among transudates.Other causes included congestive cardiac failure (09%), nephrotic syndrome (06%),& anaemias (04%). Exudative ascites was caused by tuberculosis in 10% cases &malignancy in 06% cases. Cholesterol level of >70 mg/dL was found in 82% ofmalignant ascites. So, it was concluded that ascitic fluid cholesterol levels weresignificantly greater in malignant cases in comparison to ascites caused by non -malignant aetiologies. Inference - Cholesterol concentration in ascitic fluid helps todifferentiate ascites in malignancy, from non - malignant ascites.CONCLUSIONSCholesterol estimation in ascitic fluid is a better parameter to differentiatemalignant ascites from non - malignant ascites.

Bacillus spp. for suppression of eggplant bacterial wilt pathogen in Andaman Islands: Isolation and characterization

Bacillus spp. isolated from different soils of Andaman Islands was characterized for antimicrobial potential by different methods. Among 65 strains tested, six Bacillus strains showed best in vitro antagonistic potential against solanaceous bacterial wilt pathogen Ralstonia solanacearum. The identity of the Bacillus strains were confirmed using 16S rRNA gene sequencing and biolog based phenotypic fingerprinting. The functional analysis revealed that most of the strains were positive for the production of IAA, siderophore and phosphate solubilization. In glass house and field studies, Bacillus amyloliquifaceans strain (Bs_Abi) showed significant biocontrol efficacy (92.1 and 73.7%, respectively) against brinjal bacterial wilt disease. The antimicrobial potential of Bs_Abi were further ascertained by presence of nine antimicrobial peptide (AMP) gene biosynthesis in PCR amplifications which confirmed the presence of peptide genes for six antibiotics (iturin, bacillomycin D, mycosubtilin, surfactin, bacilysin and subtilin). Our study revealed that the utilization of such antagonistic Bacillus spp. with multi antimicrobial peptide genes and more functional traits from tropical/ extreme soils would play a vital role in management of eggplant bacterial wilt disease and in formulation of new bioinoculants for improving the cropping system in Andaman and Nicobar Islands

Red cell alloimmunization in multitransfused chronic renal patients on haemodialysis in a tertiary care centre of Jammu region

Background: There is a high incidence of alloimunization in many patients with diseases that require repetitive blood transfusions. One such group is chronic renal failure patients as majority of them have severe anemia due to deficiency of erythropoietin. As many such patients are unable to afford erythropoietin, they are treated with blood transfusions. This study was thus undertaken to study alloimunization in such patients at our center.Methods: A total of 96 patients found eligible were enrolled in this cross-sectional study that was carried out from June 2016 to august 2016. After detailed history, antibody screening was done by using Immucor Panoscreen three vial set and if screening came out to be positive antibody identification was then done by using Immucor Panocell-10.Results: 96 patients including 76 male and 20 female patients recieved a total of 912 transfusions. Alloantibodies were detected in a total of 12 patients (12.5%). Of these 12 patients 8 patients had a single antibody whereas 4 patients developed two antibodies. The antibodies developed at a rate of 1.8% per transfusion (16/912). On alloantibody type identification, the most common type found was anti E (4/16) followed by both anti D and anti C in 3 patients each.Conclusions: Alloimmunization to minor erythrocyte antigens occurs in many patients of chronic renal failure. This results in frequent pre-and post-transfusion complications. Inclusion of antibody screening test in routine pretransfusion testing protocol for the patients who are at higher risk of alloimmunization and require long-term transfusion dependence is desirable.

MCQ Supplementation in a Physiology Didactic Class: A Learning Tool.

Background and Objectives: Medical educators are facing the challenge to develop innovative creative material so as to engage the students in active learning, more so for basic sciences. In addition the innovations should help student learning, keeping time and manpower and economy constraints in mind. Overall objective of this study was to study and assess the role of MCQ supplementation in a didactic class in improving student learning. Methods: 136 First year MBBS students of a government medical college attending physiology classes were recruited for the study. Based on the university recognized syllabus the Specific learning objectives (SLOs) were randomly grouped into two categories, that is, SLOs for which didactic classes were to be supplemented with multiple choice questions (MCQs) and those SLOs for which didactic classes were to be taught without MCQ supplementation. Results: On subjecting individual student’s scores obtained in the two categories of MCQs to unpaired ‘t’ test the difference was found to be statistically significant, p = 0.025, t = 2.259. 95% CI. Conclusion: Reinforcement of Didactic class with MCQ supplementation is an effective learning tool which was well received by the participants.

Exogenous application of putrescine at pre-anthesis enhances the thermotolerance of wheat (Triticum aestivum L.).

Antioxidant enzymes, besides being involved in various developmental processes, are known to be important for environmental stress tolerance in plants. In this study, the effect of treatment of 2.5 mM putrescine (Put), heat stress (HS -42°C for 2 h) and their combination on the expression and activity of antioxidant enzymes was studied at pre-anthesis in the leaves of two wheat (Triticum aestivum L.) cultivars — HDR77 (thermotolerant) and HD2329 (thermosusceptible). We observed that 2.5 mM Put before HS significantly enhanced the transcript levels of superoxide dismutase (SOD), catalase (CAT), cytoplasmic and peroxisomal ascorbate peroxidase (cAPX, pAPX) in both the cultivars. However, the activities of antioxidant enzymes (SOD, CAT, APX and GR), as well as accumulation of antioxidants (ascorbic acid and total thiol content) were higher in HDR77 than in HD2329 in response to the treatment 2.5 mM Put + HS. No significant change was observed in the proline accumulation in response to HS and combined treatment of 2.5 mM Put + HS. A decrease in the H2O2 accumulation, lipid peroxidation and increase in cell membrane stability (CMS) were observed in response to 2.5 mM Put + HS treatment, as compared to HS treatment alone in both the cultivars; HDR77 was, however, more responsive to 2.5 mM Put + HS treatment. Put (2.5 mM) treatment at pre-anthesis thus modulated the defense mechanism responsible for the thermotolerance capacity of wheat under the heat stress. Elicitors like Put, therefore, need to be further studied for temporarily manipulating the thermotolerance capacity of wheat grown under the field conditions in view of the impending global climate change.

An experimental study on the effect of isoniazid on the efficacy, plasma concentration and toxicity of paracetamol in Albino rats.

Background: Paracetamol is used for symptomatic treatment of fever and pain with isoniazid and other anti-tubercular drugs in patients of tuberculosis. Literature has conflicting data regarding their interaction. Some studies show that isoniazid increases oxidative metabolism of paracetamol whereas some other suggest that isoniazid has an inhibitory effect. The present study was conducted to find out the possible interaction between paracetamol and isoniazid. Methods: The study was undertaken on Wistar strain of Albino rats. Group I and Group II animals were treated with paracetamol (500 mg/kg) and the combination of paracetamol (500 mg/kg) and isoniazid (30 mg/kg) respectively for 2 months. Blood samples were taken before and during the study for biochemical and histopathological studies of liver and renal functions and plasma paracetamol concentration was also evaluated. Results: Isoniazid decreased the plasma paracetamol concentration without affecting its analgesic activity. However, the hepatotoxic and nephrotoxic effects of paracetamol were found to be further aggravated by isoniazid co-administration. Conclusion: Isoniazid potentiates the hepatotoxic and nephrotoxic effects of paracetamol possibly due to hepatic enzyme induction by isoniazid.

Characterization of differentially expressed stress-associated proteins in starch granule development under heat stress in wheat (Triticum aestivum L.).

Abiotic stress causes abrupt increase in the expression of stress-associated proteins, which provide tolerance by modulating the defense mechanism of plants. Small heat shock proteins (sHSPs) and anti-oxidant enzymes are important for environmental stress tolerance of the plants. In this study, two full-length cDNAs encoding small heat shock protein (sHSP) and superoxide dismutase (SOD), designated as TasHSP and SODI were identified and characterized from C-306 (thermotolerant) and PBW343 (thermosusceptible) cultivars of wheat (Triticum aestivum L.). An alpha crystalline domain was observed in TasHSP and manganese/iron binding domain in case of SODI. Quantitative real-time PCR showed very high transcript level of TasHSP and SOD in C-306 compared to PBW343 at different stages of growth and against differential heat stress (HS). Under differential HS at milky-dough stage, the fold change in transcript of both TasHSP and SOD was observed maximum in C-306, compared to PBW343. Protein profiling and isoenzymes analysis showed the expression of several heat-stable proteins and prominent isoenzymes of SOD in C-306, compared to PBW343. Scanning electron microscopy (SEM) of starch granules showed globular, well-shaped and more numbers of endospermic cells in C-306, compared to defragmented, irregular shaped and shrunken granules in case of PBW343 under HS treatment (42°C for 2 h). Diurnal change in soluble starch synthase (SSS) activity showed an increase in the activity during afternoon (35°C), compared to morning (29°C) and evening (32°C) in both the cultivars. Under heat stress (42°C for 2 h), a drastic decrease in the SSS activity was observed, due to the thermal denaturation of the enzyme. Thermotolerance capacity analyzed using cell membrane stability (CMS) showed significantly higher CMS in case of C-306, compared to PBW343 at different stages of growth. Findings suggest that abundance of TasHSP and SODI during milky-dough stage plays a very important role in starch granule biosynthesis. The mechanism may be further exploited to develop tolerant wheat cultivar with high quality seeds.

Genome Wide Identification of Target Heat Shock Protein90 Genes and Their Differential Expression against Heat Stress in Wheat.

Aims: To study the genetic and transcript profiling of the genes specifying cytosolic HSP90s in Triticum aestivum. Study Design: Random sampling. Place and Duration of Study: Indian Agricultural Research Institute, New Delhi, India, between August to December, 2011. Methodology: We include C-306 (thermotolerant) and PBW343 (thermosusceptible) cultivars of wheat for the study. Total RNA was isolated using Trizol method and gene was identified and isolated using RT-PCR. In silico characterization was done using different bioinformatic tools. Quantitative real time PCR was carried out using BioRad CFX96 platform and Pfaffl’s method was used for the comparative change in fold expression of the gene. Results: Here, we report cloning of an HSP90 gene from C-306 wheat cultivar having an ORF of 700 amino acids. Genome Blast identified 3 different clusters of reference sequence on chromosome no 4, 8 and 9 having LOC 100125696 and showing maximum homology with HSP90 reported from Triticum aestivum. Pure amino acid composition revealed highest composition of glutamic acid followed by lysine and leucine whereas, cysteine composition was lowest. Protein characterization showed the existence of 10 networks of coevolved amino acids. Quantitative real time PCR showed 1.5, 4.5, 5 & 7.4 fold increase in expression of HSP90 in case of C-306 compared to 2.5, 6.4, 6.9 & 5.6 fold increase in case of PBW343 at vegetative (root & shoot), pollination and milky dough stage. Multiple co-chaperones of HSP90 were observed by immunoblot assay in response to differential heat shock. Conclusion: This investigation proves that HSP90 is one of the key components of defense mechanism in wheat against heat stress which requires the formation of cochaperone complexes with HSP70 for its functional activity. There is a need to exploit the transcription factors associated with HSP90 and its regulation and differential expression in order to use it for developing thermotolerant wheat cultivars.