Evaluation of telomerase activation in head and neck cancer.

During replication of the linear chromosomes, telomeres, i.e. the ends of the chromosomes, are not replicated completely by the conventional DNA polymerases. Therefore, normal somatic cells senesce after certain number of cell divisions. Telomerase is a special reverse transcriptase used by most eukaryotes to achieve immortalization. Telomerase activity has been determined in a variety of cancers. However, there are few reports on telomerase activity in head and neck cancer. The etiology of the disease in India is completely different from Western countries. Tobacco consumption is more prevalent in India and the mode of tobacco consumption (e.g. chewing, snuffing, bidi smoking, reverse smoking) is also different. The present study determined telomerase activity in 32 malignant tumour samples of head and neck cancer patients, 11 samples from patients with precancerous/benign lesions and 30 samples of adjacent normal tissues. Telomerase was found to be activated in 80% of the patients with head and neck cancer, 100% of the patients with precancerous/benign lesions and 74% of the adjacent normal tissues. According to the theory of field cancerization, carcinogenic insults (e.g. tobacco) may result into multiple malignant foci. This fact may explain the reason for high telomerase positivity in adjacent normal as well as precancerous/benign tissues. Telomerase activation and the clinical or histopathological characteristics of the head and neck cancer patients were observed to be independent features. This is a preliminary report which has generated a greater interest for in-depth elucidation of the role of telomerase and telomeres in head and neck carcinogenesis in India.

Role of sialic acid and alkaline DNase in breast cancer.

Serum levels of sialic acid and alkaline DNase (ADA) were analysed in 495 blood samples collected from 170 breast cancer patients before and during/after anticancer treatment. Fifty-six healthy females were included in the study to define the cutoff values. The markers were analysed by highly sensitive spectrophotometric methods. Statistical evaluation of the data was done using Student's 't' test, paired 't' test and ROC curve analysis. The total sialic acid (TSA), lipid bound sialic acid (LSA) and ADA in sera of untreated breast cancer patients were significantly higher than in controls. ROC curve analysis revealed TSA and LSA to be useful markers for diagnosis of breast cancer. Serum levels of TSA and LSA were significantly decreased in complete responders as compared to their pretreatment values. The pretreatment ADA values showed much individual variation. However, responders showed higher levels of ADA than untreated patients. In nonresponders the values of the biomarkers were comparable with pretreatment levels. The study suggested that TSA and LSA can be helpful in the diagnosis of breast cancer. All three markers can be used for assessment of response to anticancer treatment in breast cancer patients.

Utilization of acrylonitrile by bacteria isolated from petrochemical waste waters.

A bacterium, utilising acrylonitrile as a sole source of carbon and nitrogen, was isolated from Indian Petrochemical Corporation Limited (IPCL) waste waters and identified as Arthrobacter sp. This strain could also utilize acetonitrile, acetamide and acrylamide individually as a source of carbon and nitrogen. The metabolic studies with the whole cells indicated the sequential conversion of the nitrile to the respective amide and then to the respective acid and ammonia. The rate of nitrile hydrolysis was slower than the corresponding amide hydrolysis. Acrylic acid, the end product of acrylonitrile breakdown, did not support the growth when provided as a carbon source.

In vitro effect of ethanol on polyphosphoinositide metabolism in bulk isolated brain cells.

Metabolism of polyphosphoinositide was studied in bulk isolated brain cells. Cells were isolated by a rapid method using mechanical disruption followed by molecular seiving and centrifugation. Incorporation of [32Pi]orthophosphate into phosphatidyl inositol-4-phosphate and phosphatidylinositol-4,5 bis-phosphate was optimum at 30 and 60 min, respectively, in the isolated cells. Breakdown studies showed maximum loss of 32Pi after 60 min. Addition of ethanol at and above 10 mM concentration in vitro significantly decreased the incorporation of 32Pi into both phosphatidylinositol-4-phosphate and phosphatidylinositol-4,5 bis-phosphate by the isolated cells. However, the spontaneous breakdown of polyphosphoinositide was not altered in the presence of ethanol in vitro.

Purification and properties of alpha-galactosidase from Klebsiella Sp. No. PG-2.

alpha-Galactosidase has been purified from Klebsiella Sp. No. PG-2, a bacterium isolated from rat small intestine, using calcium phosphate gel, DEAE-cellulose column chromatography and gel filtration technique. About 130-fold increase in specific activity was observed, the pH optimum of 6.5-7.0 characterizes the enzyme as neutral alpha-galactosidase. The optimum temperature was 37 degrees C and the energy of activation was 11,856 cal/mole. Km values obtained for raffinose, mellibose, stachyose and p-nitrophenyl-alpha-D-galactopyranoside were 20.0, 6.6 33.3 and 4.0 mM respectively. The activity was inhibited by p-CMB; iodoacetate, Ag2+, Hg2+, Cu2+, Pb2+ and galactose. Examination of the enzyme activity indicated that the enzyme is cytosolic and is inducible in nature.

Phytase from Klebsiella Sp. No. PG-2: purification and properties.

A phytase (EC 3.1.3.8) was extracted from rat intestinal bacterium, Klebsiella Sp. No. PG.-2, and purified 50-fold by ammonium sulphate fractionation, ion-exchange chromatography and gel filtration. The enzyme is inducible in nature. The pH optimum was at 6.0 for all the inositol phosphates studied and this characterized the enzyme as an acid phosphohydrolase. Of a range of potential substrates tested, only p-nitrophenyl phosphate alongwith the inositol phosphates was hydrolyzed. It exhibits a Km of 2.0 mM; temperature optimum of 37 degrees C and energy of activation 9,120 cal/mole for all the inositol phosphates studied. The activity was inhibited by Ag2+, Hg2+, Cu2+, fluoride and high substrate concentration.

Clinical usefulness of alterations in sialic acid, sialyl transferase and sialoproteins in breast cancer.

Sialic acid, the end moieties of the carbohydrate chains are biologically important and essential for functions of glycoconjugates and are reported to be altered in cancer patients. Two hundred and twenty five breast cancer (BC) patients, 100 patients with benign breast disease (BBD) and 100 healthy females (controls) were enrolled for the study. Eight hundred and twenty four follow-up samples of 225 breast carcinoma patients were also evaluated. The association of sialic acid forms, sialyltransferase and α-2-6 sialoproteins levels with presence and extent as well as prognosis of breast carcinoma was studied. Serum sialic acid forms and sialyltransferase revealed significantly elevated levels among untreated breast cancer patients as compared to the controls, patients with BBD as well as cancer patients in remission. Non-responders showed comparable levels of the markers with those found in breast cancer patients at the time of diagnosis. Higher levels of sialic acid forms at diagnosis were associated with poor prognosis. A positive correlation between serum levels of different forms of sialic acids and extent of malignant disease was observed. The changes in serum proteins with terminal α-2-6 sialic acid correlated well with alterations in the levels of sialic acid forms and sialyltransferase. Malignant tissues showed elevated levels of sialic acid and sialyltransferase as compared to surrounding normal tissues.The results suggested potential utility of these markers in evaluation of clinical outcome.

Ascorbic acid concentration of human fetal tissues in relation to fetal size and gestational age.

1. Studies were carried out on the distribution of ascorbic acid in human fetal tissues with the progress of gestation. 2. Fetuses and stillborn babies varying in gestational age from 12 to 38 weeks were obtained from various Baroda hospitals. Ascorbic acid levels were determined in selected tissues: brain, adrenal, liver, kidney, lung, heart and placenta. 3. Ascorbic acid concentration in the brain was higher than that in the adrenal at all gestational ages, suggesting the importance of this vitamin in brain development. The concentrations of this vitamin in liver, kidney, lung and placenta were comparable, but that in the heart tended to be lower. In all the tissues, there was a fall in ascorbic acid during late gestation. However, the levels in tissues of stillborn babies were higher than those reported for adults.

Role of microorganisms in improving nutritional status of the poor.

The intestinal microflora as well as bacteria involved in fermentation of foods ordinarily consumed in India may contribute substantially for the improvement of nutritional status of poor Indians.

Evaluation of serum sialic acid, sialyltransferase and sialoproteins in oral cavity cancer.

OBJECTIVE: Altered glycosylation of glycoconjugates is among the important molecular changes that accompany malignant transformation. The aim of our study was to investigate clinical usefulness of circulatory levels of sialic acid, sialoproteins and sialyltransferase for early diagnosis and management of oral cavity cancer (OC) patients. MATERIALS AND METHODS: Blood samples collected from 210 untreated OC patients, 100 patients with oral precancerous conditions (OPC) and 100 healthy males. OC patients were followed after initiation of anticancer treatment and 394 follow-up samples were also collected. Serum sialic acid levels were measured spectrophotometrically. Sialyltransferase activity was analysed using radioassay. Alpha 2-6 sialoproteins were isolated using lectin affinity chromatography. RESULTS: Serum levels of free, protein bound and total sialic acid as well as their ratio with total proteins were significantly elevated in untreated OC patients compared with healthy individuals, patients with OPC as well as complete responders (CR). Levels of the markers were comparable between untreated OC patients and non-responders. We observed positive correlation between serum levels of the markers and extent of malignant disease. Serum sialyltransferase activity showed significant elevations in OC patients compared with the controls (P<0.001), patients with OPC (P<0.05) and CR (P<0.05). Higher sialic acid levels in OC patients at the time of diagnosis showed poor survival. The changes in serum proteins with terminal alpha 2-6 sialic acid correlated well with the alterations in the levels of sialic acid forms and sialyltransferase activity. CONCLUSION: Our results confirmed the elevations in sialic acid and sialyltransferase levels in OC patients and suggested potential utility of these parameters in prognostication and treatment monitoring of this neoplasm. The alterations in these parameters in circulation were in accordance with the changes in alpha 2-6 sialylated proteins.