Optimisation Of Biosurfactant Production By Serratia Rubidaea Kap Isolated From Oil Contaminated Soil Sample

Surfactants are surface active molecules which play an important role in variousindustrial processes and product formation due to their interfacial properties, biodegradabilityand eco-friendly nature. Many of the chemical surfactants in use today are produced fromnon-renewable petrochemical feedstocks, hence bio surfactants produced by microorganismsusing renewable and cheap substrates are considered as viable alternatives to petroleumbased surfactants. In the current study, the optimization of various parameters for maximumproduction of bio surfactant, by Serratia rubidaea KAP isolated from oil contaminated soilsample, was carried out. Optimum bio surfactant production was obtained in a mediumcontaining; (g %) KH2PO4 (0.3), Na2HPO4 (0.6), yeast extract (0.5), NaCl (0.5), (NH4)2SO4 (0.3),MgSO4.7H2O (2mg), FeSO4.7H2O (0.002), and CaCl2.2H2O (0.006), rice bran oil (5% v/v) with C:N and C: P ratio as 8:1 and 7:1 respectively. The media was adjusted to pH 7.2, inoculated with5% v/v test culture prepared at 0.8O.D600nm, and incubated at 30°C for 96h under agitationspeed of 160rpm. The resulting bio surfactant yield of 1.0g% was achieved, with 0.88g% weightof dry biomass, 56.78% decrease in surface tension of the medium and emulsification index of87.5%. Thus the isolate showed good potential as a bio surfactant-producing bacterium thatcan be used for large scale production using rice bran oil as a cost-effective substrate, as analternative to chemical surfactants.

Utility Of Squash Cytology In Diagnosing Cns Lesions

BACKGROUND: The different modalities used for intraoperative consultation of CNS lesions are squash smear cytology, frozen sections & fluid cytology. The intraoperative diagnosis of squash cytology enables the surgeon to take decision regarding treatment. Squash technique requires small sample size, needs minimal technical equipment, is less expensive and gives better morphological detail. The present study was taken to assess the diagnostic accuracy of intraoperative squash cytology of CNS lesions. MATERIALAND METHODS: It is a prospective study conducted for one and half year in the department of Pathology. All lesions of brain and spinal tract comprised the study group. The stain used for squash cytology was rapid haematoxylin and eosin. The squash diagnosis was compared with gold standard histopathology. RESULTS: Total 96 cases were studied. Male to female ratio was 1.13:1. The overall concordance rate was 89.13% (including both complete and partial agreement cases). Good correlation was noted in diffuse astrocytoma (100%) and least correlation was noted in anaplastic oligodendroglioma (28.5%). CONCLUSION: Squash cytology is a rapid and simple technique for intraoperative diagnosis with high overall accuracy.

Quantitative & qualitative analysis of endothelial cells of donor cornea before & after penetrating keratoplasty in different pathological conditions.

Background & objectives: Endothelial cells of the donor cornea are known to be affected quantitatively and qualitatively in different pathological conditions after penetrating keratoplasty (PK) and this has direct effect on the clarity of vision obtained after PK. This study was undertaken to analyze the qualitative and quantitative changes in donor endothelial cells before and after PK in different pathological conditions. Methods: A prospective investigational analysis of 100 consecutive donor corneas used for penetrating keratoplasty between June 2006 and June 2008, was conducted. The patients were evaluated on the first day, at the end of first week, first month, third and six months and one year. Results: A decrease was observed in endothelial cell count in all pathological conditions. After one year of follow up the loss was 33.1 per cent in corneal opacity, 45.9 per cent in acute infective keratitis (AIK), 58.5 per cent in regrafts, 28.5 per cent in pseudophakic bullous keratopathy (PBK), 37 per cent in descemetocele, 27 per cent in keratoconus and 35.5 per cent in aphakic bullous keratopathy (ABK) cases. Interpretation & conclusions: The endothelial cell loss was highest in regraft cases which was significant (P<0.05), while the least endothelial cell loss was seen in keratoconus cases. The cell loss was associated with increase in coefficient of variation (CV), i.e. polymegathism and pleomorphism. Inspite of this polymegathism and pleomorphism, the clarity of the graft was maintained.

Solid-pseudopapillary neoplasm of the pancreas: A clinicopathological and immunohistochemical study of 33 cases from a single institution in Southern India.

Background: The clinicopathological and immunohistochemical data of solid-pseudopapillary neoplasm (SPN) from India are limited. Our objectives were to evaluate various histopathological parameters and immunomarkers to elucidate the best possible immunomarker combination that can accurately diagnose these tumors. Materials and Methods: Clinicopathological features of 33 consecutive cases of SPN were retrospectively analyzed. Immunohistochemistry (IHC) was performed on a tissue microarray of 31 of these cases with antibodies to pan-cytokeratin (CK), vimentin, CD 10, E-cadherin, β-catenin, estrogen receptor (ER), progesterone receptor (PR), chromogranin. Results: Totally, 32 of 33 patients were females with a mean age of 26 years (range 12-62 years). Majority presented with abdominal pain (93.93%) and/or abdominal mass (48.48%). Location in pancreas included tail (14), head (7), body and tail (6), body (5) and neck (1). The mean diameter of the tumor was 8.6 cm (range 0.5-16 cm). Surgical exploration was done in all cases. On IHC, tumor cells were consistently positive for vimentin and negative for chromogranin (100%). CK was positive in 12 cases (38.7%), CD 10 showed perinuclear dot positivity in 14 (45.16%) cases and cytoplasmic positivity in 05 (16.12%) cases. All cases showed loss of membranous staining for both β-catenin and E-cadherin with nuclear positivity in 90.32% and 70.16% cases, respectively. PR was positive in 20 (64.51%) cases, while all of them were negative for ER. Conclusion: Solid-pseudopapillary neoplasm is a tumor commonly affecting young females. Loss of membrane expression of β-catenin and E-cadherin with nuclear positivity can be used confi rm the diagnosis of SPN.

Production and Partial Characterization of Alkaline Protease from Bacillus Tequilensis Strains CSGAB0139 Isolated from Spoilt Cottage Cheese.

An alkaline protease producing strain was isolated from spoilt cottage cheese sample which was identified as Bacillus tequilensis strain SCSGAB0139 on the basis of morphological, cultural, biochemical characteristics and 16S rRNA sequence analysis. Primary screening for protease production was carried out by observing for zone of hydrolysis on skim milk agar, GYEA milk agar and gelatin agar plates. Physicochemical parameters like pH of the medium, incubation time and temperature, aeration and composition of the medium were optimized for maximum protease production by this isolate. Maximum yield of protease (85.67U/ml) was obtained in a medium containing peptone (5% w/v), maltose (5% w/v) and KNO3, 0.5%; K2HPO4, 0.4%; trisodium citrate, 0.4; CaCl2, 0.0002%; MgSO4·7H2O, 0.05%; Na2CO3, 1%.; 1% (v/v) of a trace element solution (NH4)6MO7O24, 0.01%; FeSO4·7H2O, 0.2%; CuSO4·5H2O, 0.02%; ZnCl2, 0.02%) having pH 10, inoculated with 1%(v/v) of pre-grown cell mass and incubated at 30°C on a rotary shaker (100rpm) for 48hrs. Absence of any one of the following salts viz. KNO3, K2HPO4, tri-sodium citrate; MgSO4, CaCl2 and Na2CO3 from optimized medium reduced the protease production by 80% to 40%. The enzyme has an optimum pH of 9 and maintained its stability over a broad pH range between 6 and 10. Its optimum temperature is 30°C, and exhibited a stability of up to 65°C. Among metal ions only Ca2+ and Mg2+ions enhanced the enzyme activity up to 105% and 107% respectively while other metal ions reduced the activity by 40% where as EDTA exhibited the least inhibitory effect upon the enzyme. Protease activity was enhanced in the presence of isopropanol and marginally reduced in the presence of other organic solvents studied. The crude enzyme showed stability towards various surfactants such as Tween-20, Tween- 80, SDS and Triton X-100. It also showed excellent stability and compatibility with commonly used laundry detergents (Ariel, Surf excel and Surf Blue). The enzyme retained its activity in 2% H2O2 indicating it to be bleach-stable. The present findings show that the protease from Bacillus tequilensis strain SCSGAB0139 is alkali- and detergent- stable and hence, when this protease was applied to remove blood stains from cotton fabrics indicated its potential use in detergent formulations.

Comparative Morphological and Anatomical Study on Thymus Gland of Human and Primate.

Context: The comparative morphological and anatomical study on thymus was carried out in human and primate. The prenatal stage of Macaca radiata was selected for the present study. Study Design: Cross sectional analytical type of study. Place and Period of study: Department of Anatomy, Dr. A.L.M. PG Institute of Basic Medical Sciences, Chennai from July 1999 to June 2000. Materials: The comparative morphology and anatomy of thymus of human embryonic, 10 weeks, 15 weeks and prenatal foetuses, and monkey foetus was carried out. Methods: Comparative micro-anatomical study was done by paraffin processing method. The sections were stained as per the method published by Culling (1974). Results: In monkey foetus, the thymus gland is slightly elongated, whereas in human foetuses it is not elongated and oval in shape. The size of the thymus is larger in human foetuses than monkey foetus. In both cases cells are parenchymal in nature. Due to spatial organization in human foetuses, the lymphocytes aggregation is more in cortex than in medulla. In monkey foetus the lymphocyte aggregation is simpler in arrangement through spatial organization is much less.

Sodium iodide symporter, estrogen receptor, and progesterone receptor expression in carcinoma breast - An immunohistochemical analysis.

Background: Sodium iodide symporter (NIS), a transporter of iodine is essential for thyroid hormone biosynthesis. It also plays a role in the radioiodine treatment of thyroid cancers. NIS mediated radioiodine transport to breast cancers is under active investigation due to its potential therapeutic utility. Cellular localization and quantification using immunohistochemistry may provide clues for its utility in management of carcinoma breast. Materials and Methods: Human NIS (hNIS) expression was therefore assessed by utilizing a rabbit polyclonal antibody raised against a cloned hNIS in different grades of infiltrating duct carcinoma of breast and its metastatic deposits namely in lymph nodes, bone marrow, and endometrium. Further, hNIS expression was compared with prognostic markers namely estrogen receptor (ER) and progesterone receptor (PR). Results: hNIS was positive in 90.6% cases (29/32) and Scarff-Bloom-Richardson grading was done in 25 cases and 23 cases were NIS positive. Among nongraded cases, 2/2 cases of carcinoma in-situ were positive and 4/5 were positive in cases having post therapy residual tumor status. The strong positivity for hNIS was seen irrespective of ER or PR status and of grade of breast carcinoma and correlated well with western blot analysis. In all the three metastatic sites, NIS was positive in the tumor. Conclusion: These findings indicate the utility of immnohistochemistry for NIS as a new potential prognostic marker and may provide guidance for possible radio iodine therapy in breast cancer patients.

Fungal infections as a contributing cause of death: An autopsy study.

Context: With the continuing rise in the number of immunocompromised patients, the incidence of invasive mycoses has increased. Various studies have reported the trends of fungal infections in autopsies. Because of limitations in antemortem clinical diagnosis owing to lack of sensitive diagnostic tools, information regarding frequency and pathogenesis of fungal infections is largely dependent on autopsy studies. Aim: To study the prevalence of fungal infections at autopsy spanning a period of 20 years and to document recent trends, prevalence of various fungi over decades along with underlying predisposing factors and pathological findings. Settings and Design: Retrospective study. Materials and Methods:All autopsies between 1988 and 2007 were reviewed and all cases showing fungal infections were analyzed. The clinical details and demographic data were retrieved from medical records. Representative sections from all organs were stained with hematoxylin and eosin stain and special stains including Gomori's silver methenamine (GMS) and per-iodic acid Schiff (PAS). Culture details were noted, wherever available. Results: A total of 401 autopsies were performed during the study period. Fungal infections were identified in 35 (8.7%) of these cases. Leukemia was the commonest risk factor. The commonest pathogen in the present study was Aspergillus sp. The commonest single organ involved was brain (n = 18). Culture positivity was seen in 23.8% cases. Conclusion: The study highlights various predisposing factors and organisms in autopsy series. Existing diagnostic modalities are not sensitive to ensure antemortem diagnosis of fungal infections.

Curbing the growth of candida-A comparative in-vitro study.

Endodontics is the least forgiving branch of dentistry in terms of success or failure. Therefore, re-treatment has become a significant part of today’s endodontic practice. There is a growing evidence showing the presence of fungi especially Candida albicans in root canals of obturated teeth in which endodontic treatment has failed. It has been demonstrated that Candida species are resistant to some medications used in endodontics. The aim of this study was to examine the in-vitro susceptibility of Candida albicans to materials used in endodontics such as mineral trioxide aggregate (MTA), Calcium hydroxide Ca(OH)2/camphorated paramonochlorophenol (CMCP) /glycerine and chlorhexidine (CHX). MTA and CHX are effective anti-fungal agents each with 67% anti fungal activity while Ca(OH)2/CMCP showed 33% anti-fungal activity.

Light chain immunofluorescence in various nephropathies.

Context: Light chain immunofluoresence (IF) in renal biopsy is routinely used in the diagnosis of light chain deposition disease (LCDD), amyloidosis and cast nephropathy. Light chain predominance has also been reported in certain glomerulopathies like IgA nephropathy. However, pathogenesis of this pattern of deposition in various glomerulopathies is uncertain. Aim: To discuss the pathogenesis and utility of light chain IF in nephropathies. Setting and Design: Retrospective study. Materials and Methods: The pattern of light chain IF and light microscopic diagnosis in 306 cases of various nephropathies was reviewed. Direct IF was done in all these cases with commercial fluorescence (Fluoresciene Isothiocynate ) conjugated polyclonal rabbit anti-human antisera against IgM, IgG, IgA, C3, C1q, kappa and lambda light chains. Results: Light chain deposits were seen in 240 (78.43%) cases. In IgA nephropathy, lupus nephritis and post-infectious glomerulonephritis (PIGN), lambda positivity was more as compared to kappa. Light chain deposits in LCDD and membranous nephropathy were more kappa type. The IF pattern in amyloidosis was not consistent. Conclusion: The pathogenesis of light chain predominance in glomerulopathies is not clear and it depends on isoelectric point and size of the immune complex. Light chain IF should be performed routinely in all the renal biopsies.

Morphologic evaluation of renal function using semi-quantitative method in primary nonproliferative glomerular diseases.

Context: Fibrosis is universally accepted as a poor prognostic finding in renal pathology. Semi-quantitative assessment is widely used for prognostication in pathology. Aims: We propose a semi-quantitative method to prognosticate primary nonproliferative glomerular diseases. Settings and Design: A semi-quantitative method based on Banff schema, 97 classification has been modified to suit the requirements. Glomerular, tubulointerstitial, and vascular compartments were scored independently, and the scores were totaled to obtain total scores. Materials and Methods: Seventy-six renal biopsies were assessed by semi-quantitative scores and the individual compartmental and total scores were correlated with serum creatinine levels. Follow-up was available in 24 cases. Statistical Analysis: Pearson correlation coefficient, two-tailed t test, to determine the P value. Results: P values were significant for the total scores as well as individual compartments. There is a linear correlation between the scores and serum creatinine levels. A total score of ≥5 was significant. Conclusions: The semi-quantitative scoring system based on modified Banff schema, 1997 is useful in prognosticating renal biopsies in primary nonproliferative glomerular diseases.

Salivary duct carcinoma: Correlation of morphologic features by fine needle aspiration cytology and histopathology.

Background: Salivary duct carcinoma (SDC) is a highly aggressive primary salivary gland neoplasm that resembles intraductal and infiltrating breast carcinoma. Objectives: To review cytomorphologic features of histology proven SDC and evaluate potential pitfalls in cytologic diagnosis. Materials and Methods: Fine needle aspiration cytology (FNAC) of five histologically proven SDCs were reviewed. Results: One patient was an elderly male (61 years), while the other four patients were younger, in their fourth decade (average age: 38 years). The initial cytologic diagnoses in two of the cases were poorly differentiated carcinoma with differential diagnosis of SDC and high grade mucoepidermoid carcinoma, while in the third case, a possibility of malignant mixed tumor was suggested. In fourth and fifth cases, the diagnosis was suggestive of pleomorphic adenoma with cystic change. The spectrum of cytologic changes included flat sheets and cohesive papillary and three-dimensional clusters. There was moderate to severe nuclear pleomorphism and atypia. Cribriform pattern and necrosis were occasionally identified. Prominent bright granular metachromatic stroma was seen in two of the cases interpreted as pleomorphic adenoma with cystic change and in the tumor reported as suggestive of malignant mixed tumor. The fifth case showed numerous cyst macrophages and apocrine cells with mild nuclear atypia. Conclusion: FNAC of SDC is difficult to interpret because of overlapping cytomorphologic features. Bland cytomorphologic features in some cases and several clinical pitfalls are demonstrated in our series.

Gill lesions in the major carp, Labeo rohita exposed to lethal and sublethal concentrations of tannery effluent.

The major carp, Labeo rohita were exposed to (0.873%) lethal and sublethal (0.073%) concentrations of tannery effluent for 24h and 40 days respectively under static bioassay condition. The surface architecture of gill revealed severe damages such as : fusion and clumping in the middle and distal parts of the primary lamellae, swelling and deterioration of the cells. The interlamellar space was filled either with hyperplastic epithelial or mucous cells. Secondary lamellae lost their identity and appeared finger like in structure in the lethal concentration and necrosis was observed in the primary and secondary epithelium. Swelling of primary and secondary epithelial cells was evident in sublethal concentration.

Identification of a hypothetical membrane protein interactor of ribosomal phosphoprotein P0.

The ribosomal phosphoprotein P0 of the human malarial parasite Plasmodium falciparum (PfP0) has been identified as a protective surface protein. In Drosophila, P0 protein functions in the nucleus. The ribosomal function of P0 is mediated at the stalk of the large ribosomal subunit at the GTPase centre, where the elongation factor eEF2 binds. The multiple roles of the P0 protein presumably occur through interactions with other proteins. To identify such interacting protein domains, a yeast two-hybrid screen was carried out. Out of a set of sixty clones isolated, twelve clones that interacted strongly with both PfP0 and the Saccharomyces cerevisiae P0 (ScP0) protein were analysed. These belonged to three broad classes: namely (i) ribosomal proteins; (ii) proteins involved in nucleotide binding; and (iii) hypothetical integral membrane proteins. One of the strongest interactors (clone 67B) mapped to the gene YFL034W which codes for a hypothetical integral membrane protein, and is conserved amongst several eukaryotic organisms. The insert of clone 67B was expressed as a recombinant protein, and immunoprecipitaion (IP) reaction with anti-P0 antibodies pulled down this protein along with PfP0 as well as ScP0 protein. Using deletion constructions, the domain of ScP0, which interacted with clone 67B, was mapped to 60-148 amino acids. It is envisaged that the surface localization of P0 protein may be mediated through interactions with putative YFL034W-like proteins in P. falciparum.

Plant products as protective agents against cancer.

Out of various spices and leafy vegetables screened for their influence on the carcinogen-detoxifying enzyme, glutathione-S-transferase (GST) in Swiss mice, cumin seeds, poppy seeds, asafoetida, turmeric, kandathipili, neem flowers, manathakkali leaves, drumstick leaves, basil leaves and ponnakanni leaves increased GST activity by more than 78% in the stomach, liver and oesophagus, - high enough to be considered as protective agents against carcinogenesis. Glutathione levels were also significantly elevated in the three tissues by these plant products. All of them except neem flowers, significantly suppressed (in vivo) the chromosome aberrations (CA) caused by benzo(a)pyrene in mouse bone marrow cells. Multiple CA and exchanges reflecting the severity of damage within a cell were significantly suppressed by these nine plant products. The results suggest that these nine plant products are likely to suppress carcinogenesis and can act as protective agents against cancer.