Short term response of plankton community to nutrient enrichment in central eastern Arabian Sea: Elucidation through mesocosm experiments.

Oligotrophic waters (OW), generally favour longer food chain facilitated by the microbial loop. In such ecosystems, physical mixing (e.g. upwelling, and winter convection) inject nutrients and propagules from subsurface to the photic zone. Such events are expected to alter the food chain through shifts in the plankton community. Mesocosm experiments were carried out to evaluate the influence of nutrient enrichment from the deep (100-150 m) on the surface plankton community for the first time in the Arabian Sea, through custom-designed enclosures in OW of the central-eastern Arabian Sea (CEAS). Surface water was characterized by low nutrients and phytoplankton biomass (chlorophyll-a of <0.2 µg m-3) and upon nutrient enrichment yielded differing response. Higher abundance of picophytoplankton, bacteria and protists was noticed at a depth of ~100 m than at surface. The inoculation of such a population to the surface, resulted in a significant enhancement of autotrophic (picophytoplankton) and heterotrophic (bacteria and protists) populations. However, significant changes in the abundance of larger plankton was not evident till three days of incubation. Even though autotrophic picophytoplankton responded positively, a distinct increase in chlorophyll-a was not evident. This study points out that the lack of sufficient viable microphytoplankton propagules, neither at the surface nor at the depth (inoculum) are the possible reasons for the lack of their distinct positive response. These experiments suggest the dominance of microbial community response to physical mixing in the OW regions of the Arabian Sea and the importance of propagule diversity. The insights from this experiment will serve as a precursor for appropriate modifications in ocean modelling and forecasting studies and help in building global environmental management tools.

A nomogram for predicting the risk of neck node metastasis in pathologically node-negative oral cavity carcinoma.

OBJECTIVE: To generate a nomogram for predicting the risk of neck node metastasis in pathologically node-negative patients using a combination of variables comprising of protein expression, ultrastructural alterations and clinicopathological parameters. MATERIALS AND METHODS: Surgically removed oral tumours (n = 103) were analysed for the expression of desmosomal and hemidesmosomal assembly proteins by immunohistochemistry and ultrastructural alterations by transmission electron microscopy (TEM). Protein expression, ultrastructural alterations and clinicopathological variables were used to construct nomogram from the training set in 75 patients. Clinical utility of the nomogram was validated in a discrete set of 28 patients. RESULTS: Univariate and multivariate analyses were performed on the training set, and obtained significant variables comprising of integrin ß4 expression (p = .027), number of hemidesmosomes (p = .027)/desmosomes (p = .046), tumour differentiation grade (p = .033) and tumour thickness (p = .024) were used for construction of the nomogram. The area under the curve was calculated for both training 0.821 (95% CI 0.725-0.918) and validation sets 0.880 (95% CI 0.743-1.000). The nomogram demonstrated a predictive accuracy of 73.3% and 78.6% with the sensitivity of 81.4% and 83.3% in the training and validation sets, respectively. CONCLUSIONS: The nomogram constructed on postsurgical tumour samples will be a value addition to histopathology for the detection of neck node metastasis in pathologically node-negative patients.

Assessment of heavy metal contamination of road dusts from industrial areas of Hyderabad, India.

Road dust in industrial areas carries high levels of toxic heavy metals. Exposure to such polluted dust significantly affects the health of people residing in these areas, which is of major concern. The present study was taken up with an aim to highlight the magnitude and potential sources of accumulation of heavy metals in 32 road dust samples collected from six industrial areas of Hyderabad. Acid-digested sample solutions were analyzed by ICP-MS for Cu, Zn, Cr, Co, Pb, Ni, V, Zr, Ce, Y, and Hf. The road dusts exhibit significantly high mean metal levels which are much above their crustal abundances. The relative ordering of mean metal contents is Zr > Zn > Pb > Cr > Ce > Cu > V > Ni > Y > Co > Hf. Elevated pollution indices (I geo, EF, C (i) f, and C deg) reveal that the road dusts are pollution impacted showing varying degree of heavy metal contamination. Strong positive correlations exhibited by metal pairs Cu-Zn, Cr-Ni, Ce-V, Y-Ce, and Hf-Zr imply their origin from common anthropogenic sources. Principal component analysis grouped the metals according to the sources which contributed to their accumulation. The present study confirms to an intensive anthropogenic impact on the accumulation of heavy metals in the studied road dusts attributable mainly to strong influences of vehicular and industrial activity and partly to domestic and natural processes. The results obtained imply the need for further investigations to assess their ecological implications and human health risks.

Molecular phylogenetic profiling of gut-associated bacteria in larvae and adults of flesh flies.

Flesh flies of the genus Sarcophaga (Diptera: Sarcophagidae) are carrion-breeding, necrophagous insects important in medical and veterinary entomology as potential transmitters of pathogens to humans and animals. Our aim was to analyse the diversity of gut-associated bacteria in wild-caught larvae and adult flesh flies using culture-dependent and culture-independent methods. Analysis of 16S rRNA gene sequences from cultured isolates and clone libraries revealed bacteria affiliated to Proteobacteria, Actinobacteria, Firmicutes and Bacteroidetes in the guts of larval and adult flesh flies. Bacteria cultured from larval and adult flesh fly guts belonged to the genera Acinetobacter, Bacillus, Budvicia, Citrobacter, Dermacoccus, Enterococcus, Ignatzschineria, Lysinibacillus, Myroides, Pasteurella, Proteus, Providencia and Staphylococcus. Phylogenetic analysis showed clone sequences of the genera Aeromonas, Bacillus, Bradyrhizobium, Citrobacter, Clostridium, Corynebacterium, Ignatzschineria, Klebsiella, Pantoea, Propionibacterium, Proteus, Providencia, Serratia, Sporosarcina, Weissella and Wohlfahrtiimonas. Species of clinically significant genera such as Ignatzschineria and Wohlfahrtiimonas spp. were detected in both larvae and adult flesh flies. Sequence analysis of 16S rRNA gene libraries supported culture-based results and revealed the presence of additional bacterial taxa. This study determined the diversity of gut microbiota in flesh flies, which will bolster the ability to assess microbiological risk associated with the presence of these flies. The present data thereby establish a platform for a much larger study.

Clinical significance of aberrant vimentin expression in oral premalignant lesions and carcinomas.

OBJECTIVE: To investigate the clinical significance of vimentin expression at early and late events of tobacco/areca nut-associated oral tumorigenesis. MATERIALS AND METHODS: Immunohistochemistry (IHC) was carried out on paraffin-embedded tissues of oral mucosa normal (n = 10), inflammatory lesions (n = 19), leukoplakia (n = 52), submucous fibrosis (n = 71) and tumours/cut margins (n = 227 each), using anti-vimentin antibody, and the expression profile was correlated with patients' clinical parameters. Immunofluorescence, Western blot and RT-PCR analysis were also carried out wherever adequate and fresh tissues were available. RESULTS: Aberrant vimentin expression was seen in hyperplastic, dysplastic and fibrotic tissues, which showed statistically significant correlation with the histopathological grade of dysplasia (P = 0.001) and fibrosis (P = 0.009). Vimentin expression also showed statistically significant correlation with tumour size (P = 0.048), clinical stage (P = 0.013), regional lymph node metastases (P = 0.001), local recurrence (P = 0.001) and survival (P = 0.021) of patients with oral squamous cell carcinoma (OSCC). Its expression in invasive fronts statistically correlated with development of nodal metastasis and local recurrence. CONCLUSIONS: Our results suggest possible role of vimentin in early events of tobacco/areca nut-associated oral tumorigenesis, which may prove useful to predict the malignant potential of high-risk oral lesions. Further, association between vimentin expression in invasive fronts and aggressive phenotype of tumours may help clinicians to choose the appropriate treatment modality for OSCC management.

Mumbai harbour, India: gateway for introduction of marine organisms.

Ships have been identified as one of the important vectors in the translocation of organisms from one bioregion to another leading to bioinvasion. In this context, harbours serve as a gateway for the introduction of alien species. Surveys were carried out in the vicinity of ports of Mumbai for macrobenthic fauna, zooplankton and hard substratum community on three different occasions during 2001-2002. The study shows that 14 polychaete species are recently introduced to this area. Mytilopsis sallei, a bivalve, which is an invasive species in the Indian context continued to be present but was restricted to enclosed docks, indicating preference for embayed water bodies. The polychaete Protula tubularia was abundant in the hard substratum community and is being reported as a possible ship-mediated introduction.

Eutrophication status of marine environment of Mumbai and Jawaharlal Nehru ports.

The marine environment of Mumbai and Jawaharlal Nehru ports was monitored for some environmental and biological parameters during three different periods between 2001 and 2002. The results are compared with the records available since 1960s. With the passage of time the environmental status underwent changes, probably due to the increase in anthropogenic activities in the metropolis. The nutrient level especially the nitrate concentration has increased gradually over the years with a simultaneous decrease in dissolved oxygen, indicating increase in the biological activity. Characterization of this environment based on Assessment of Estuarine Trophic Status (ASSETS) model indicates that the current status is poor and may get worsen in future if no appropriate management policies are put into place.

Cytokeratin expression in oral submucous fibrosis--an immunohistochemical study.

BACKGROUND: Oral submucous fibrosis (OSF) is a pre-malignant condition caused by habitual use of areca nut, affecting the oro-pharynx and characterized by progressive fibrosis. Alteration of cytokeratin (CK) expression has been documented in leukoplakia and oral cancer (OC). However, very little is known of CK alterations in OSF. The present study was carried out to characterize the CK profile in OSF and ascertain if this could be used as a surrogate marker for malignant transformation. METHODS: Paraffin-embedded tissues of OSF (n = 50), normal (n = 10) and OC (n = 10) were stained with pancytokeratin (PanCK), high molecular weight cytokeratin (HMWCK), CKs 18, 14, 8, 5, 4 and 1 by immunohistochemistry. RESULTS: Significant difference in the CK staining pattern was seen between normal, OSF and cancer. Significant changes in OSF included increased intensity of staining for PanCK and HMWCK, aberrant expression of CK8 and decreased expression of CKs 5 and 14. CONCLUSION: Cytokeratin profile of OSF was significantly different from normals for PanCK, HMWCK, CK8, 5 and 14 suggesting their potential to be used as surrogate markers of malignant transformation. Further studies will help in better defining the nature and clinical implications of these alterations.

Gyrineum natator: a potential indicator of imposex along the Indian coast.

Imposex related studies have used neogastropods as sentinel organism. In this effort, we elucidate the potential of a mesogastropod, Gyrineum natator, for imposex monitoring on the central west coast of India. Observations were made with specimens collected from Mumbai (three stations), Ratnagiri (two stations), Goa (one station) and Mangalore (one station). The analyzed populations at all four sites were affected by imposex. The six different stages of imposex in this organism are illustrated through schematic and scanning electron micrographs. In view of the anticipated ban on TBT based antifouling paints this baseline information can serve as an indicator for environmental monitoring in this region.

Demonstration of cytokeratin-5 non-expression in tobacco related oral carcinogenesis--use of reverse transcriptase polymerase chain reaction as a sensitive assay.

Cytokeratins (CK) are the epithelia specific intermediate filament proteins. We have shown consistent non-expression of CK-5 protein in human oral pre-cancer and cancer, in earlier studies. To investigate whether non-expression of CK-5 protein is the result of transcriptional or translational block and to evaluate the possibility if CK-5 non-expression can be used as a marker for early diagnosis of tobacco related oral cancer, RT-PCR using CK-5 specific primers was conducted. Out of 36 precancerous lesions and 29 squamous cell carcinomas (SCC) of buccal mucosa (BM) samples studied, 11 and 13 samples respectively of precancer and SCC did not show CK-5 product in RT-PCR. Down regulation of CK-5 mRNA expression was also observed in some samples. Thus, in conclusion, our results have shown that CK-5 non-expression is the result of transcriptional block. We proposed CK-5 non-expression as a potential marker for the early diagnosis of tobacco related oral cancer.

Effect of omega-3 fatty acid (docosahexanoic acid) on BRCA1 gene expression and growth in MCF-7 cell line.

Recently, we have demonstrated that omega-6 fatty acid linoleic acid (LA) in presence of estradiol (E2) enhances proliferation and anchorage independent growth with down regulation of BRCA1 mRNA expression in MCF-7 cell line. Since omega-3 fatty acid (docosahexanoic acid, DHA) is known to block the promoting effect of omega-6 polyunsaturated fatty acid (LA), we wanted to see whether addition of DHA can inhibit the growth of MCF-7 cells which are exposed to LA + E2 and any alteration of BRCA1 mRNA expression could be seen in DHA treated culture. Experiments on MCF-7 cells with DHA revealed both decrease in proliferation and anchorage independency as compared to controls; while no change of BRCA1 mRNA expression was observed. Further, when DHA was administered to cells along with LA + E2, no change in BRCA1 expression was observed, however, a marked decrease in proliferation and soft agar colony formation was evident, indicating inhibition of MCF-7 cells following DHA treatment. Flow cytometric analysis showed that DHA treated cells either alone or in combination with LA + E2 induced marked G1/S and G2/M arrest of the cells, suggesting the inhibitory effect of DHA at this phase of cell cycle. However, neither typical DNA ladder nor fragmented nuclei or apoptotic bodies were observed, ruling out presence of apoptosis following DHA treatment.

Cytokeratin expression in human fetal tongue and buccal mucosa.

Expression of cytokeratins (CK), a subset of intermediate filament (IF) proteins in epithelia, is developmentally regulated. CK expression may also change after malignant transformation. Our earlier studies on CK expression in human oral tumours and pre-cancerous lesions have shown specific changes in CK expression. We analysed CK expression in human tongue and buccal mucosa (BM) in fetuses in the embryonic age group of 16 to 27 weeks using biochemical and immunohistochemical techniques to find out whether there is any similarity in CK expression in human oral squamous cell carcinomas (SCC) and fetal oral tissues. CK 1, 8 and 18 were detected in a majority of samples using both techniques. Our earlier studies had shown aberrant expression of CK 1 and 18 in many of the oral SCC and leukoplakias. Studies by immunohistochemistry showed that these different CK antigens were expressed in different cell layers. CK 1(2) were present in the stratified epithelial layers whereas CK 8 and 18 were restricted to glandular epithelium. Till 27 weeks of gestation, both tongue and BM expressed CK 1, 8 and 18 along with CK 6 and 16. Thus, fetal tissues showed some similarities in CK pattern with their respective SCC.

Role of vitamins C and E as chemopreventive agents in the hamster cheek pouch treated with the oral carcinogen-DMBA.

OBJECTIVE: To evaluate the role of vitamins C and E as chemopreventive agents in oral carcinogenesis by optical and ultrastructural studies. MATERIALS AND METHODS: The cheek pouch of male hamsters was treated with the oral carcinogen, dimethylbenz(a)anthracene (DMBA), to induce multiple tumour formation. Vitamins C and E were applied either singly or in combination as a chemopreventive agent. Paraffin and resin-embedded sections of the hamster cheek pouch were studied optically and ultrastructurally. RESULTS: The epithelium of control hamsters showed hyperorthokeratosis and parakeratosis, but did not develop well differentiated squamous cell carcinoma (WDSCC). Ninety percent of the animals treated with DMBA alone showed WDSCC while 10% of the animals developed papillomas. There was also a marked increase in the number of cells undergoing mitosis in this group. A reduction in the yield (1.1 tumour/animal) and rate 60-80% of squamous cell carcinomas but not of papillomas (2.0 papillomas/animal) was observed in groups VI-VIII treated with DMBA and vitamins C and E singly or in combination as compared to those of DMBA only. In animals treated with DMBA plus vitamins C and E, statistical significant decrease in the number of animals with tumours and mitotic basal cells was observed when compared with the DMBA treated group. Control animals showed normal ultrastructural morphology while tumour-bearing animals showed basal lamina in a discontinuous, fragmented, broken and diffused basement membrane, with diminished lamina densa, fewer hemidesmosomes and invagination of the basal cell cytoplasmic processes in the subepithelium. CONCLUSION: These results indicate that vitamin E singly or in combination with vitamin C plays a role in the inhibition of tumour cell growth.

Ultrastructural changes to the palatal mucosa of rats following the application of 4-nitroquinoline-1-oxide (4NQO) and vitamin C.

Oral mucosal neoplastic disease in rodents has been induced by various chemical carcinogens, including water soluble 4-nitroquinoline-1-oxide (4NQO). It has been suggested that vitamin C can inhibit, delay or prevent the development of neoplasms, as well as enhance the induction of neoplasia. In this investigation, 4NQO was used to produce a high yield of carcinomas of the palatal mucosa of rats in a relatively short period of time and topical vitamin C was applied to delay the neoplastic transformation. The temporal aspects of the ultrastructural changes occurring in 4NQO-induced oral palatal mucosa treated with both 4NQO and vitamin C have been described and discussed.

Cytokeratin expression in precancerous lesions of the human oral cavity.

Cytokeratin (CK) expression was studied in buccal mucosa (BM) from 20 leucoplakia and 7 submucous fibrosis patients using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), immunoblotting and two-dimensional gel electrophoresis with iso-electric focussing (IEF) as the first dimension. Normal BM expresses CK 4, 5, 13, 14 and perhaps 19. Of 20 leucoplakia samples analysed, CK 5 was not detected in 17 samples, while CK 14 was not found in 13 samples. CK 1 and CK 8 were aberrantly expressed in six and seven samples, respectively. CK expression in contralaterally collected uninvolved tissues from 3 patients showed a normal pattern in two samples. Non-expression of CK 5 was observed in five of seven submucous fibrosis samples, while CK 14 was not detected in only two samples. CK 8 was aberrantly expressed in three samples. All the leucoplakia patients were chronic tobacco chewers. Thus, non-expression of CK 5 may be an early event occurring in tobacco-associated pathological changes in the BM.

Subchronic oral hepatotoxicity of turmeric in mice--histopathological and ultrastructural studies.

Dietary administration of the whole spice turmeric (0.2%, 1.0%, 5.0%) or ethanolic turmeric extract (ETE, 0.05%, 0.25%) for 14 days, at doses reported to be cancer preventive in model systems, were found to be hepatotoxic in mice. Histopathological evaluation showed coagulative necrosis accompanied by a zone of regenerating parenchymal cells of liver. The ultrastructural changes in liver parenchymal cells were non-specific reaction to injury. Results suggest mouse to be a susceptible species for turmeric induced toxicity.

Consumption of 'country liquor' and its relation to alcoholic liver disease in Mumbai.

The amount of alcohol intake required for the development of liver disease has been determined in Western populations; corresponding figures in Indians, many of whom consume locally brewed liquors, are not known. We studied 328 patients from a public hospital in Mumbai who admitted to regular alcohol consumption, to determine the pattern of alcohol consumption and its relation to liver disease. Liver disease was more common in those who consumed illicitly-brewed as compared to licit liquor. Daily drinking, volume of consumption > 200 ml per day, and duration of drinking > 14 years were each significantly more common in those with liver disease. A cumulative intake of > 2000 ml. years, calculated as the product of volume (ml per day) and duration (years), was a reliable cut-off level for association with liver disease (sensitivity 65%, specificity 77%) and cirrhosis (sensitivity 70%, specificity 59%). The content of alcohol in these liquors, estimated in 23 samples, ranged from 23-36.1 g/100 ml, being lower in the illicit liquors. Thus, in Mumbai, alcoholic liver disease occurs more commonly with consumption of illicit liquor (despite its lower alcohol content); liver involvement appears earlier and with lower consumption levels than in the West.

Spectrophotometric estimation of rhenium with 5-chloro-2-hydroxythiobenzhydrazide after its extraction separation using n-octylaniline.

A solution of n-octylaniline in chloroform extracts rhenium selectively from 0.12-0.16 M nitric acid medium. Rhenium from the organic phase is backstripped with 5% aqueous ammonia solution and estimated spectrophotometrically with 5-chloro-2-hydroxythiobenzhydrazide (5-Cl-2-OHTBH). 5-Cl-2-OHTBH forms a blue complex with rhenium on heating in the acidity range 0.40-3.5 M HCl. The complex is extracted into chloroform and its absorbance is measured at 580 nm. The complex is stable for more than 12 h with a molar extinction coefficient of 9.0 x 10(3)l mole(-1) cm(-1) and a Sandell sensitivity of 0.019 mug cm(2).

The effect of vitamin C on the hamster cheek pouch treated with the water soluble carcinogen 4-nitroquinoline-1-oxide (4NQO).

Vitamin C is an essential nutrient whose protective influence is carcinogenesis has been reported frequently, suggesting that vitamin C inhibits the formation of some carcinogens and decreases the incidence and delays of the neoplastic lesions. However, the mechanisms by which this occurs are unknown. In this study, the water soluble carcinogen 4-nitroquinoline-1-oxide (4NQO) has been used to induce a high yield of tumours in the oral cavity either singly or in combination with tobacco. Since the mucosa of rats is less susceptible to carcinogens than the hamster cheek pouch, the hamster cheek pouch has been used to study the influence of vitamin C on 4NQO-induced oral malignancy. The aim of this study was to determine whether topically applied vitamin C had an effect on the oral carcinogenesis induced by application of 4NQO. Similarly, an attempt was made to study the modulating effect of vitamin C on the histopathological and ultrastructural changes during the neoplastic process in the hamster. Vitamin C appeared to delay tumour induction and had other protective effects against neoplasia.

Chemical characterization of exopolysaccharides from the marine fouling diatom amphora coffeaeformis.

Amphora coffeaeformis cells were grown in batch cultures under continuous illumination at 18°C for 10 d. Algal cells were removed by centrifugation, lyophilized and used for the extraction of exopolysaccharides using either 0.05 M EDTA, 1 M NaOH or 1.5 M NaCl. The 1.5 M NaCl treatment removed most exopolysaccharides. Glucose (81%) was the most abundant monosaccharide in the exopolysaccharides. The chemical composition data suggest that the exopolymers were acidic sulphated polysaccharides containing high concentrations of pyruvate (22%) and uronic acids (18%). These polysaccharides may play an important role in metal complexation and protection from desiccation in A. coffeaeformis.