Biocomputational identification and validation of novel microRNAs predicted from bubaline whole genome shotgun sequences.

MicroRNAs (miRNAs) are small (19-25 base long), non-coding RNAs that regulate post-transcriptional gene expression by cleaving targeted mRNAs in several eukaryotes. The miRNAs play vital roles in multiple biological and metabolic processes, including developmental timing, signal transduction, cell maintenance and differentiation, diseases and cancers. Experimental identification of microRNAs is expensive and lab-intensive. Alternatively, computational approaches for predicting putative miRNAs from genomic or exomic sequences rely on features of miRNAs viz. secondary structures, sequence conservation, minimum free energy index (MFEI) etc. To date, not a single miRNA has been identified in bubaline (Bubalus bubalis), which is an economically important livestock. The present study aims at predicting the putative miRNAs of buffalo using comparative computational approach from buffalo whole genome shotgun sequencing data (INSDC: AWWX00000000.1). The sequences were blasted against the known mammalian miRNA. The obtained miRNAs were then passed through a series of filtration criteria to obtain the set of predicted (putative and novel) bubaline miRNA. Eight miRNAs were selected based on lowest E-value and validated by real time PCR (SYBR green chemistry) using RNU6 as endogenous control. The results from different trails of real time PCR shows that out of selected 8 miRNAs, only 2 (hsa-miR-1277-5p; bta-miR-2285b) are not expressed in bubaline PBMCs. The potential target genes based on their sequence complementarities were then predicted using miRanda. This work is the first report on prediction of bubaline miRNA from whole genome sequencing data followed by experimental validation. The finding could pave the way to future studies in economically important traits in buffalo.

Raised serum homocysteine levels in patients of coronary artery disease and the effect of vitamin B12 and folate on its concentration.

Homocysteine(Hcy) has been implicated as a novel risk factor of Coronary Artery Disease (CAD) among Asian Indians, but many studies done in India failed to reveal any direct correlation. It has also been reported that Folic acid and Vitamin B12 levels inversely affect serum levels of homocysteine. In this study, we looked at the levels of homocysteine among patients with CAD. The effect of Vitamin B12, Folate and other risk factors on homocysteine levels were also evaluated. Mean homocysteine levels in cases (22.81±13.9, n=70) were significantly higher (p=<0.001) than the controls (7.77±7.3, n=70). However no statistically significant correlation could be deduced between homocysteine Vitamin B12, and Folate. Cumulative analysis have indicated an increase in homocysteine levels among patients with CAD with every additional risk factor.

Improvement in fresh frozen plasma transfusion practice: results of an outcome audit.

Blood components have been in use in clinical practice for many decades now. In spite of fairly clear guidelines regarding their use, inappropriate prescriptions for components are still rampant. We undertook this work to assess the appropriateness of fresh frozen plasma (FFP) transfusions in our hospital. A prospective audit of 504 transfusion orders for 1761 FFP units was conducted over a 6-month period which was followed by a re-audit of 294 FFP prescriptions for 961 units. In the initial audit, we identified 304 (60.3%) prescriptions which were inappropriate according to the British Committee for Standardization in Hematology (BCSH) guidelines. The re-audit performed after an educational campaign among clinicians showed a reduction in inappropriate requests by 26.6%. The specific areas of misuse were FFP transfusions in patients with hypoproteinaemic states (40.5%), anaemia (36.5%), bleeding without coagulation factor deficiency (10.2%) and volume depletion (9.2%). A significant 50.3% of requests in the initial audit and 38.4% in the re-audit were for single- or two-unit transfusions, which were subtherapeutic. FFP transfusions carry the same risks to the patients as any other blood component. Prescribers of these transfusions need to be aware of the clinical setting where their use is appropriate. Local hospital transfusion committees can play a vital role in overseeing transfusion practices to ensure optimal use of blood/component therapy.

A multicentric study to evaluate efficacy and safety of cefetamet pivoxyl in lower respiratory tract infections in Indian patients.

This multicentric, open label, non-comparative study was designed to evaluate the extended spectrum of third generation oral cephalosporin, cefetamet pivoxyl in the treatment of patients with lower respiratory tract infections. This study was conducted among 111 patients with clinical, radiological and bacteriological findings consistent with the diagnosis. After obtaining written informed consent, patients were given cefetamet 500 mg tablet twice a day for 7 days. Cefetamet consistently decreased all clinical signs and symptoms at post-therapy visit. All the treated patients were either cured or improved. Cefetamet was well tolerated with a low incidence of drug related adverse events. The findings of this study indicate that cefetamet pivoxyl was well tolerated and is suitable option for the treatment of patients with lower respiratory tract infection.

Correlation of Braunwald's clinical classification of unstable angina pectoris with angiographic extent of disease, lesion morphology and intra-luminal thrombus.

One hundred consecutive patients (81 male and 19 female) with unstable angina pectoris undergoing coronary angiography were divided according to Braunwald's clinical classification. Seventeen (17%) patients had new onset angina (class I), 68 (68%) sub-acute angina (class II) and 15 (15%) had acute rest angina (class III). Twenty-seven (27%) patients had secondary unstable angina pectoris (class A), 49 (49%) primary unstable angina (class B) and 24 (24%) had post-infarction unstable angina (class C). ST-T wave changes on ECG were present in 54 (54%) while absent in 46 (46%) patients. On coronary angiography, 26 (26%) patients had single vessel disease, 30 (30%) double vessel disease and 39 (39%) patients had triple vessel disease. Five (5%) patients were found to have normal coronaries. Classification of patients according to Braunwald's clinical classification showed single vessel disease to be higher in class I as compared to class II (47% vs 22%; p = 0.04) and classes III (47% vs 20%; p<0.01). Single vessel disease was found to be higher in class C as compared to class B (41.7% vs 16.4; p = 0.01). Double vessel disease was higher in class B as compared to class A (40.8% vs 18.5%, p = 0.04). Triple vessel disease incidence was not found to be significantly different among different clinical classes. Morphology of coronary artery lesions was classified according to Ambrose's classification. Out of the total of 248 lesions in the whole study group, there were 68 (27.42%) concentric lesions, 55 (22.18%) eccentric type I lesions, 23 (9.27%) eccentric type II lesions, 42 (16.94%) multiple irregularity lesions and 60 (24.19%) totally occluded lesions. Concentric lesions were found to be higher in class C as compared to class B (40% vs 19.8%; p = 0.014). Statistically significant difference was not present in the distribution of other morphological type of lesions among different clinical classes. In the whole study group, intra-luminal thrombus was found to be present in 17 (17%) of patients. Distribution of intra-luminal thrombus according to Braunwald's classification showed that none of the patients in class I had intra-luminal thrombus, while 13 (19.1%) patients in class II and 4(26.7%) in class III had intra-luminal thrombus. The difference in the occurrence of intra-luminal thrombus between class I and class II (p = 0.004) and class I and class III (p = 0 .03 was found to be significant. Thus, majority of patients undergoing coronary angiography had primary sub-acute rest angina. Single vessel disease was higher in new onset angina. Patients with unstable angina pectoris and ST-T changes on ECG had higher number of lesions per patient and higher eccentric type I lesions. Intra-luminal thrombus was more frequently encountered with acute rest angina. However, the distribution of different morphological type of lesions on coronary angiography did not differ significantly in different clinical classes of unstable angina pectoris divided according to Braunwald's classification.

Cholecystectomy under regional anesthesia in a patient with total Kartagener's syndrome.

A 41-year-old woman with complete Kartagener's syndrome developed cholecystitis in a left-sided gall bladder. Bilateral diffuse bronchiectasis ruled her unfit for general anesthesia; chronic cough, copious expectoration and dyspnea prevented her from lying down supine for more than a few minutes. Successful cholecystectomy was performed in a semi-reclined position using thoracic epidural anesthesia, the anatomy being the mirror image of that of a gall bladder in normal position. The gall bladder was full of stones and operative cholangiogram revealed a normal but transposed biliary system.

Angiographic severity and morphological spectrum of coronary artery disease in non insulin dependent diabetes mellitus.

Coronary angiographic profile of 75 patients (63 males and 12 females) with noninsulin dependent diabetes mellitus (NIDDM) and CAD was compared with 75 nondiabetic patients (63 males and 12 females) with CAD. No difference was present between the mean age (56.2 +/- 7.4 vs 56.1 +/- 7.7 years; p = NS), presenting complaints (67 unstable angina and 8 stable angina with positive TMT in both the groups) and other coronary risk factors between the two groups. Severity and diffuseness of coronary artery involvement was assessed by a coronary artery score (CAS) using the segmental distribution method for coronary artery lesions. Diabetic patients with CAD had a higher CAS (18.7 +/- 10.3) as compared to the nondiabetic patients with CAD (12.7 +/- 9.6) (p < 0.01). Diabetic patients with CAD had a higher number of TVD [43 (57.3%) vs 31 (41.3%); p < 0.01] while the DVD and SVD was not significantly different. As compared to the nondiabetic group, diabetics had a higher total number of coronary artery lesions (300 vs 200; p < 0.001), a higher lesion per patient ratio (4.0 lesions/patient vs 2.6 lesions/patient; p < 0.001), a higher number of concentric lesions, [151 (50.3%) vs 90 (45%); p < 0.01] and a higher number of multiple irregularity lesions, [36 (21%) vs 27 (9%); p < 0.05]. The diffuse involvement of vessels was not significantly different between the two groups in LAD (12.1% vs 5.3%; p = NS), LCx (14.2% vs 5.8%; p = NS) and RCA (10.5% vs 5.0%; p = NS).(ABSTRACT TRUNCATED AT 250 WORDS)

Neurocryptococcosis in acquired immunodeficiency syndrome.

Two cases of Neurocryptococcosis in AIDS are described. The first patient presented with generalised lymphadenopathy, hepatosplenomegaly and raised intracranial tension. The clinical manifestations of second patient included prolonged fever and hepatosplenomegaly. Both the patients had evidence of disseminated cryptococcosis. The distinctive clinical manifestations, investigative abnormalities and modification of therapy of neurocryptococcosis manifestations in AIDS are discussed.

Reaching the child in need.

"Reaching the Deprived Child" is India's theme for the Year of the Child. This paper describes the recording system employed in the Community Health Programme of Christian Medical College, Ludhiana and how it enables us to reach not only deprived children, but those in danger of deprivation, the majority being the children of the socioeconomically underprivileged. Many are nutritionally normal at 12 months but 57 per cent of the males and 75 per cent of the females suffer from malnutrition in the 2nd year, 50 per cent of females having severe malnutrition. The etiology, discerned from a study of the deprived, allowed a preventive approach to be designed for those at risk. Intensive nutrition education is given in the homes, commencing as each baby reaches 5 months, and mothers are taught to feed them on suitable modifications of the food available in Punjab homes. The results in respect of 123 babies who had a minimum of 6 months of this care, are described. Complete compliance was achieved in 82 per cent and in the rest partial compliance was achieved. The nutritional status of these infants, compared with a control group, improved greatly, especially in females in whom the prevalence of normal nutrition rose from 26 to 61 per cent and that of severe malnutrition fell from 50 to 17 per cent. The success of this approach depends on an accurate knowledge of the community since this allows priorities to be selected with clarity and maximum effort to be expanded upon those in greatest need.