A Rapid Method to Screen for Multi Drug Resistance (MDR) Trait in Clinical Isolates of Acinetobacter baumannii-Acyl Homoserine Lactone (AHL) Screening in CRAB and CSAB Isolates

Aims: Our proposal aimed to evaluate Acyl Homoserine Lactones (AHL) as a functional marker for Multi drug resistant (MDR) potential in clinical isolates of Acinetobacter baumannii. We investigated the AHL production potential of clinical isolates using a biosensor assay directly on a commonly used agar media. Place and Duration of Study: Department of Molecular Diagnostics and Biomarkers, Gleneagles Global Hospitals, Lakdikapul, Hyderabad-500004. Methodology: Antimicrobial drug sensitivity testing (AST) was performed on 72 clinical isolates of A. baumannii against two front-line antibiotics, Imipenem (10µg) and Meropenem (10µg), by Kirby-Bauer disk diffusion method. Production of long chain Acyl Homoserine lactone (AHLs) in the clinical isolates of A. baumannii was tested by cross streaking with the biosensor Chromobacterium violaceum mutant strain CV026 and Agrobacterium tumefaciens (NTL4pZLR4) by agar plate diffusion assay. Screening and identification of the quorum sensing mediator gene abaI was done by PCR to confirm its presence in all the 72 clinical isolates. Results: Out of the 72 clinical isolates, 58 were Carbapenem resistant Acinetobacter baumannii (CRAB) and 14 were Carbapenem sensitive Acinetobacter baumannii (CSAB) for AST by agar disc diffusion method. None of our isolates produced short chain AHLs whereas all the isolates could produce varying amounts of long chain AHLs. Genotypic confirmation of AHL gene was obtained by abaI gene PCR. Conclusion: Carbapenems are the front-line antibiotics used to treat gram negative bacterial infections in emergencies and in the critical care units of hospitals. Clinical isolates A. baumannii has innate resistance to several antibiotics due to various mechanisms, biofilms forming the first line of defense against antibiotics for the bacterium. Our study used AST to carbapenem as the leading marker for MDR, assuming the innate resistance of A. baumannii to other beta lactam antibiotics. Our study brought out certain important observations namely: a) All clinical isolates of A. baumannii produced Quorum Sensing signal molecules, the AHLs b) the clinical isolates of A. baumannii did not produce any short chain AHLs b) All the clinical isolates of A. baumannii produced long chain AHLs c) AHL production is not specific to carbapenem drug resistance because even CSAB isolates produced AHL d) AHL production is inherent to all clinical isolates of A. baumannii and it apparently indicates an underlying biofilm potential and MDR trait in these A. baumannii isolates. e) AHLs could be a universal marker for revealing MDR trait and biofilm potential in clinical microbiology AST profiling protocols.

Histopathological evaluation of surgically treated adult renal tumors: Report from a tertiary care center in India.

BACKGROUND: This retrospective study was performed in a tertiary care center from India analyzing the histopathological reports and the clinical data of the adult patients admitted in this institute with a diagnosis of renal tumors and had undergone nephrectomy for the disease. OBJECTIVE: The objective of this study is to determine the relative frequencies of different renal tumors in adults (above the age of 16 years) and to analyze the histopathological characters of the tumors. MATERIALS AND METHODS: In this retrospective study, we have analyzed the histopathology reports along with the demographic and clinical data of the adult patients who had undergone nephrectomy for renal tumors in our institute from January 2005 to December 2011. RESULTS: A total 113 adult patients underwent tumor nephrectomy during the last 7 years in our institute. Mean age of the patients was 54.5 years (range 16‑69 years). Male:Female ratio was 1.9:1. Out of 131 cases of adult renal tumors, 91.6% cases were malignant and 8.45 cases were benign tumors. Among the malignant tumors, renal cell carcinoma was the most common type. There were 2 cases of renal primitive neuroectodermal tumors and one case of renal myofibroblastoma in our series. CONCLUSION: The spectrum of adult renal tumors in this series is consistent with the other series of cases reported by different authors. Only few cases of the renal tumors were diagnosed incidentally among our patients which is just opposite to the rate of renal tumors diagnosed incidentally in the developed countries. Myofibroblastoma, a benign kidney tumor diagnosed in our series is probably the first reported case in the world.

Totally implantable venous access ports: Retrospective review of long-term complications in 81 patients.

Aim of The Study: A totally implantable venous access port (TIVAP) has become an essential prerequisite for many chemotherapy protocols. It is serving its purpose very well, but its use is not without complications. We are presenting our experience with these devices (TIVAPs). Subjects and Methods: We retrospectively reviewed the totally implantable venous access ports in 81 patients at our hospital between January 2009 and March 2011 for long-term problems which include postoperative and follow-up problems, excluding the immediate complications which occur at the time of insertion. Results: Catheter malfunction was the most common complication (9.87%, 0.40/1000 device-days of use/observation). Catheter-related bloodstream infections were present in 5 (6.17%) patients (0.25/1000 device-days of use/observation). The mean life of the catheter was 246 days. Only 11.1% ports required removal during the treatment period. Overall, patients either completed treatment (82.8%) or died (6.1%) while receiving treatment. Conclusion: TIVAPs provide safe and reliable vascular access for patients on chemotherapy but require utmost care by a dedicated team of trained medical professionals and paramedics experienced with the use of such ports, in order to minimize the complications and their continued use while administering treatment.

Relation between increased anaerobic glycolysis and visual acuity in long-standing type 2 diabetes mellitus without retinopathy.

In long-standing diabetes mellitus, blood flow to essential organs including the retina is reduced owing to macrovascular and/or microvascular changes. Poor glycolytic pathway of glucose metabolism owing to tissue hypoxia caused by ischemia at capillary bed of essential organs produces excessive lactic acid and less of adenosine triphosphate, which lead to poor cellular function. The purpose of the study was to evaluate the relationship between increased anaerobic glycolysis and visual acuity in type 2 diabetes mellitus without retinopathy. Fifty patients of type 2 diabetes mellitus of 10-12 years duration, without retinopathy, constituted the study group. The controls were 50 age-matched healthy persons without diabetes mellitus. Blood lactate level and best-corrected visual acuity (BCVA) were measured in both the groups. The mean blood lactate level was 1.05 mM/l in the control group and 2.32 mM/l in the study group. BCVA of 20/20 (log MAR 0) was seen in 48 (96%) patients of the control group and in 27 (54%) patients of the study group. BCVA of 20/30 (log MAR 0.2) was seen in 23 (46%) patients in the study group and 2 (4%) in the control group. Association of higher blood lactate level with decreased BCVA in the study group was statistically significant (P< 0.001).

Desarrollo de improntas para el diagnostico del virus epstein-barr por inmunofluorescencia indirecta / Development of slides for Epstein-Barr virus diagnosis by indirect immunofluorescence

El vírus de Epstein-Barr (VEB) es el principal agente oncogénico linfotrópico dentro de la família Herpesviridae y se encuentra mundialmente distribuído. La primoinfección se produce en adultos jovenes y se manifesta como mononucleosis infecciosa. La detección de anticuerpos anti-viral cápside antigen (VCA) indica infección previa o presente com VEB. Además, se observan títulos elevados de anticuerpos anti-VCA en las enfermidades neoplásicas asociadas al VEB como los linfomas, em indivíduois HIV-positivos. El objetivo de este estúdio fue el desarrollo y puesta a punto de improntas de células P3HR1 para la detección serológica del VEB por técnicas de inmunofluorescencia indirecta (IFI). Se estimularon cultivos de células P3HR1 en crecimiento exponencial com phorbol-12-mirystoil-13-acetato y se recolectaron alícuotas a distintos tiempos para realizar improntas. Se realizó uma IFI com cada impronta usando como anticuerpo primário um suero VEB-positivo. Se observo un aumento del 11% em la expresión del VCA a las 40 horas post-estimulación, deyendo al 3.5% a las 48 horas. Estos datos fueron corroborados por ensayo de Western blot com inmunodetección. La precisión intra- e inter-lote de las improntas fue evaluada para anticuerpos IgM e IgG, com sueros probados previamente por equipos para esta determinación disponibles en el mercado para el VEB y com sueros reactivos para otros miembros de la família Herpesviridae. No se obtuvieron resultados falsos-negativos ni falsos-positivos para el VEB ni se observo reactividad cruzada com otros herpesvirus. Las improntas desarrolladas constituyen un instrumento para el diagnóstico de la primoinfección del VEB y la detección serológica de anticuerpos IgG anti-VCA de neoplasias asociadas al VEB.

Rural people and fine needle aspiration cytology (FNAC)--some findings in North Bengal Medical College, Darjeeling, West Bengal.

A hospital based interdepartmental collaborative study was carried out from 1st July to 30th September, 2000 on 269 rural people residing in six districts of the northern part of West Bengal and attending the Pathology Department in North Bengal Medical College for Fine Needle Aspiration Cytology (FNAC) after being referred from different clinical departments. The objectives of the study were to study some aspects of the outcomes of FNAC among rural people and to suggest for its wider application in rural community through Community Health Centre/Rural Hospital/Block Primary Health Centre. The results shows that Hindus (80.3%) are attending more in number than Muslims (14.13%), Christians (4.83%) and Buddhist (0.74%); 53.54% of the population are in 11-40 years age group. In benign conditions 76.95% are coming from up to 100 kms. of N.B.M.C; people in lower per capita income group of up to Rs.400/- per month are attending twice in number than those in the income groups of Rs.401/- and more per month. In malignancy however people do not think of distance or expenses due to seriousness of the diseases. In 14.5% cases FNAC remains inconclusive whereas in 85.5% cases it provides definite diagnoses.

Role of hypo-osmotic sperm swelling test in assisted reproduction.

Hypo-osmotic sperm swelling (HOS) profile is found to be a better predictor of the fertilisability in those husbands possessing subnormal semen parameters. The group where husband's HOS gradings were less than 60%, no one could conceive his fertile wife even after intra-uterine insemination (IUI) for 8 times. However, in the other group where the HOS values were greater than 60%, conception were registered in 6 oligospermic and 2 asthenospermic husbands after 2 to 8 IUI amongst 22 married couples. No conception occurred in any normal coital cycle. All the pregnancies were delivered by elective caesarean section (4 females and 3 males), their birth weights were 2.705 +/- 0.425 kg. One aborted spontaneously at 10 weeks' gestation. The emerging fact in this study is where the HOS values are below 60% the male fertility is reduced, the chances of conception are trivial but fecundity is relatively fair where HOS reading is greater than 60%. It is also a no risk and high benefit method to pick and choose subfertile husbands who are likely to conceive their fertile wives by assisted reproductive techniques.

Antitumor effect of cisplatin against murine ascites Dalton's lymphoma.

Cisplatin treatment brings about complete regression of ascites Dalton's lymphoma in mice. In the present study various steps involved during the tumor regression are examined using some biochemical and morphological parameters. During tumor regression, ascites fluid volume decreases sharply and there is an increase in carbohydrates and decrease in protein contents in the ascites supernatant after cisplatin treatment in vivo. Cisplatin treatment brings about definite changes in the arrangement/movement of surface membrane ruffles/blebs of tumor cells and causes infiltration of leukocytes (neutrophils, monocytes, lymphocytes) towards tumor cells. Membrane vesicles and vacuoles are also formed before the disintegration and lysis of tumor cells.

Jaundice in pregnancy: a clinical study.

In a study on jaundice during pregnancy it was observed that the incidence of jaundice in pregnancy was 1 in 429. Majority of the cases were primigravida, 70% of cases were between 20 and 30 years of age, 60% of the patients were unbooked, 25% of cases were with severe degree of jaundice in respect to serum bilirubin level and 70% cases were with anaemia. There were preponderance of cases in 3rd trimester. Majority of the cases (60%) had jaundice-delivery interval of above 4 weeks. Among the causes virus hepatitis was commonest one (80%). Maternal mortality was recorded as 1 in 20 cases and perinatal mortality was 7 in 17 births (41%).

Outcome of teenage pregnancy and labour: a retrospective study.

During the period of study of 3 years (1985-1987), 4698 (18.68%) cases of labour in teenage mothers were recorded out of a total of 25,142 deliveries in the obstetric unit. Preponderance of primigravida (76.6%) and cases from rural areas (51.3%) were recorded. Antenatal care was nil or inadequate in 48.6% cases. Eclampsia and pre-eclampsia affected teenage mothers (10.6%) were much more frequent than mothers of 20 years of age and above (5.2%). Incidence of 30% low birth weight baby, 20.1% prematurity and 16.4% perinatal mortality were recorded.