Molecular biology and immunology of fungal allergens

Fungi are non-chlorophyllus microorganisms, which constitutes the main source of outdoor and indoor allergens. The antigens present in the spores and fragments of hyphae induce allergic responses in sensitized patients. The frequently recognized fungi associated with asthma include Alternaria, Cladosporium, Aspergillus, and Penicillium. With the advent of molecular biology techniques a number of fungal genes encoding relevant allergens have been cloned and the expressed allergens purified and characterized. In this review, we have presented the recent developments, where recombinant allergens have been used in the precise diagnosis of fungal allergy. We have also discussed the role played by these allergens and the T- and B-cell epitopes in the immune mechanism in fungal allergy.     

Liquid–Liquid Extraction and Solid Phase Extraction for Urinary Organic Acids: A Comparative Study from a Resource Constraint Setting

Pre analytical process of extraction for accurate detection of organic acids is a crucial step in diagnosis of organic acidemias by GCMS analysis. This process is accomplished either by solid phase extraction (SPE) or by liquid–liquid extraction (LLE). Both extraction procedures are used in different metabolic laboratories all over the world. In this study we compared these two extraction procedures in respect of precision, accuracy, percent recovery of metabolites, number of metabolites isolated, time and cost in a resource constraint setup. We observed that the mean recovery from SPE was 84.1 % and by LLE it was 77.4 % (p value <0.05). Moreover, the average number of metabolites isolated by SPE and LLE was 161.8 ± 18.6 and 140.1 ± 20.4 respectively. The processing cost of LLE was economical. In a cost constraint setting using LLE may be the practical option if used for organic acid analysis.     

Congruity in Quality Indicators and Laboratory Performance

Proficiency of laboratory services is the mainstay in clinical medicine in providing error free diagnostic results. The efficiency of the laboratory needs to be evaluated as per standard international criterion. The quality indicators of the different phases of total testing process are considered the fundamental measurable tool for evaluation of laboratory performance. In order to optimize the laboratory’s proficiency and accreditate it as per international standard in our newly established lab, the study was conducted to evaluate the frequency of errors incurred by laboratory and nonlaboratory professionals during the whole testing process. Retrospective analysis was done for data received from April 2016 to Dec 2016 in our lab. Total number of samples received was 61,674, out of which 43200 samples could be analyzed for quality indicators. Total numbers of tests processed in these samples were 172,800. In the study samples, 26.5% errors were due to pre-analytical factors whereas 9.4% of errors were contributed by analytical phase and 18% by post-analytical phase. Inappropriateness of test requisition was observed to be the major attributing determinant for pre-analytical errors. Instrumentation efficiency in form of frequent breakdown (~7%), greatly affected the proficiency of analytical phase in our lab. 12% of post-analytical errors were ascribed by excessive turn-around-time. However, timeliness of critical value call out and reporting for STAT samples revealed high proficiency up to 97%. High error rates were observed in pre–pre- and pre-analytical phases that also accorded for high error frequency in post analytical phase. This emphasizes urgent need to formulate guidelines for processing all steps of total testing process and initiate strategic measures for reducing risk of errors and increasing patient safety.     

Association of hyperhomocysteinemia to alcohol withdrawal in chronic alcoholics

This study is conducted in chronic alcoholics to assess the association of an Individual’s level of serum homocysteine with the success of achieving alcohol deaddiction in the patient. The patients’ nutrition status is also assessed. 50 chronic alcoholics admitted to a deaddiction center were inducted into the study. Patients underwent an 8 weeks holistic program to promote rehabilitation from alcoholism. All the patients were addicted to alcohol for 8 – 10 years. Of the 50 patients enrolled, 39 of them completed the 8 weeks program including complete abstinence from alcohol during this period. Fasting blood samples were collected on admission and again after 8 weeks of alcohol abstinence for analyses of serum homocysteine and serum prealbumin, transferrin, total proteins and albumin, gammaglutamyl transferase (GGT) and alanine transaminase (ALT). Of the 50 patients enrolled in the study, 39 completed the 8 weeks rehabilitation program. 11 patients discontinued within 2 weeks of admission. During the 8 weeks of complete alcohol abstinence, patients were given a balanced diet and multivitamin supplements. A significant improvement in their nutritional status was noted by the elevation of serum levels of prealbumin, transferrin, total proteins and albumin. Serum homocysteine levels decreased significantly (p<0.002) to normal levels from previous hyperhomocytenemia. This was accompanied by decrease in serum GGT and ALT levels indicating improved liver functions. Serum estimation of homocysteine in chronic alcoholics is important to assess whether the patient will have a successful rehabilitation. Normal homocysteine levels are achieved after dietary changes and abstinence from alcohol. Timely correction of hyperhomocysteinemia also provides successful rehabilitation.     

Effect of Vitamin E on sP-Selectin Levels in Pre-Eclampsia

P-selectin, a cell adhesion molecule is elevated in many inflammatory conditions including preeclampsia which is characterized by generalized endothelial dysfunction and vasoconstriction presumably due to free radicals or mediators released by defective placentation. Vitamin E has been documented to protect cell membranes from oxidative damage and also decrease platelet aggregation. The role of vitamin E in pre-eclampsia is contradictory and hence the study was undertaken. Soluble P-selectin was measured by ELISA and Vitamin-E levels in plasma was estimated spectrofluorometrically. In our study the effect of supplementation of 400 IU/day of Vitamin E (a-tocopheryl acetate) to patients of pre-eclampsia showed significant decreased levels of soluble P-selectin by 2nd week as compared to patients given placebo (P = 0.005). In this short period of study no direct correlations were observed between Vitamin E or P-selectin levels with blood pressure as well as with proteinuria. Future studies may focus on the effect of a-tocopheryl acetate or the phosphate form of Vitamin-E, recently proposed to be the more active form on other inflammatory markers like IL-6, an important stimuli of P-selectin release in pre-eclampsia.     

Oxidative injury and antioxidant vitamins E and C in Schizophrenia

Susceptibility of Schizophrenic patients to lipid peroxidation relative to healthy control subjects was investigated by measuring the malondialdehyde (MDA) levels in plasma. The main finding was that Schizophrenic patients were more susceptible than control subjects to oxidative damage as evident from increased MDA levels in plasma. Antioxidant levels are also depleted in Schizophrenic patients when compared to normal subjects as evident from decreased levels of vitamins E and C in the plasma. Impaired antioxidant defense and increased lipid peroxidation suggests that treatment with antioxidants (Vitamin E, Vitamin C, beta carotene) at the initial stages of illness may prevent further oxidative injury and deterioration of associated neurological deficits in Schizophrenia.     

Role of Brain Biomarker in Predicting Clinical Outcome in Hypertensive Cerebrovascular Ischemic Stroke

This study was undertaken to evaluate the role of serum neuron specific enolase (NSE) in prediction of disability and neurological worsening in hypertensive ischemic cerebrovascular stroke. 80 hypertensive ischemic stroke patients diagnosed by a neurologist as per WHO definition along with radiological findings suggestive of cerebrovascular stroke and differentiating from hemorrhagic stroke and 60 controls having essential hypertension coming to hospital because of regular checkup or headache but with no neurological disease were included in the study. Neurological disability was assessed by NIHSS at the time of admission (within 72 h from the onset of stroke) and on 7th day after admission and cases were categorized into mild, moderate and severe disability. Venous blood samples were drawn within 72 h from the onset of symptoms. The samples were processed as per the laboratory protocol. The serum NSE samples were analyzed using an enzyme immunoassay based on the sandwich technique. We observed raised serum NSE in hypertensive ischemic stroke (17.4 ± 5.4 ng/ml) with significant association between different hypertensive groups than in hypertensive controls (9.1 ± 0.75 ng/ml). Greater degree of disability was observed in hypertensive stroke patients with raised serum NSE and hypertensive patients with mean serum NSE level of 22.9 ± 3.6 ng/ml and dyslipidemia had greater probability of neurological worsening as compared to those with mean serum NSE level of 12.7 ± 1.2 ng/ml. Serum NSE levels can serve as a peripheral indicator of neuronal damage and assist in the prediction of disability and clinical outcome in hypertensive cerebrovascular ischemic stroke patients.