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K Mehta

Publications and source records attributed to K Mehta.

At least 19 recordsLinked to original sources
PubMed2006

Tissue transglutaminase expression promotes cell attachment, invasion and survival in breast cancer cells.

Distant metastasis is frequently observed in patients with breast cancer and is a major cause of cancer-related deaths in these patients. Currently, very little is known about the mechanisms that underlie the development of the metastatic phenotype in breast cancer cells. We previously found that metastatic breast cancer cells express high levels of tissue transglutaminase (TG2), but established no direct link between TG2 and metastasis. In this study, we hypothesized that TG2 plays a role in conferring the metastatic phenotype to breast cancer cells. The results obtained suggested that increased expression of TG2 in breast cancer cells contributes to their increased survival, invasion and motility. We further found that TG2 protein in a metastatic breast cancer MDA-MB231 cells was present on the cell surface in close association with integrins beta1, beta4 and beta5. Downregulation of endogenous TG2 by small interfering RNA inhibited fibronectin (Fn)-mediated cell attachment, survival and invasion. Conversely, ectopic expression of TG2 augmented invasion of breast cancer cells and attachment to Fn-coated surfaces. We conclude that TG2 expression in breast cancer cells plays an important role in the development of the metastatic phenotype.

Breast Neoplasms↗
PubMed2006

Implications of increased tissue transglutaminase (TG2) expression in drug-resistant breast cancer (MCF-7) cells.

The development of resistance to chemotherapeutic drugs is a major obstacle to the successful treatment of breast cancer. Ways to block or overcome this resistance are objects of intense research. We have previously shown that cancer cells selected for resistance against chemotherapeutic drugs or isolated from metastatic tumor sites have high levels of a calcium-dependent protein crosslinking enzyme, tissue transglutaminase (TG2) but no direct link between TG2 and resistance was established. As TG2 can associate with the beta members of the integrin family of proteins, we hypothesized that TG2 promotes cell survival signaling pathways by activating integrins on the surface of these cells. To test this hypothesis, we studied the expression of TG2 and its interaction with various integrins in drug-resistant MCF-7 breast cancer cells. TG2 closely associated with beta1 and beta5 integrins on the surface of drug-resistant MCF-7 (MCF-7/Dox and MCF-7/RT) cells. The incubation of TG2-expressing drug-resistant MCF-7 cells on fibronectin (Fn)-coated surfaces strongly activated focal adhesion kinase, an event that leads to the activation of several downstream signaling pathways and, in turn, can confer apoptosis-resistant phenotype to cancer cells. The role of TG2 in Fn-mediated cell attachment, cell growth, and cell survival functions was further analysed by small interfering RNA (siRNA) approach. Inhibition of TG2 by siRNA-inhibited Fn-mediated cell attachment and cell survival functions in drug-resistant MCF-7 cells. We conclude that the expression of TG2 in breast cancer cells contributes to the development of the drug-resistance phenotype by promoting interaction between integrins and Fn.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed2004

Dependence of Giardia lamblia encystation on novel transglutaminase activity.

Earlier, we found that three protein disulfide isomerases (PDI) from Giardia lamblia (gPDI) also have transglutaminase (TGase) activity in vitro. We now show that differentiating Giardia cells contain isopeptide bonds (epsilon(gamma-glutamyl)lysine), the biological product of TGase activity that results in irreversible crosslinking of proteins in vivo. HPLC analyses showed the highest isopeptide bond content in cells encysting for 21 h, indicating an important role for TGase early in encystation. We were not able to detect isopeptide bonds in water-resistant cysts, possibly because they could not be extracted. One of the hallmarks of early encystation is the formation of encystation secretory vesicles (ESV) that transport nascent cyst wall proteins (CWPs) to the outer cell surface. ImmunoEM and live-cell immunofluorescence assays of encysting parasites revealed that gPDIs 1-3 are located in ESV and that gPDI-2 is also novel in that it is localized on the cell surface. Cystamine, a widely used TGase inhibitor, caused a dose-dependent inhibition of ESV formation by 21 h, thereby preventing development of trophozoites into cysts. Since cystamine (0.5-1 mM) inhibited the TGase activity of recombinant gPDIs 1-3 in vitro, PDIs appear to be the physiologic targets of cystamine. We found that when parasites were treated with cystamine, CWPs were not processed normally. These data suggest that TGase-catalyzed reactions may be needed for either the machinery that processes CWP precursors or their recruitment to ESV.

Animals↗
PubMed2002

Development of a web-based SEM specifically for K-12 education.

The scanning electron microscope (SEM) is uniquely suited for use in education due to its ability to produce clear three-dimensional-looking images of virtually any sample. Elementary and secondary science textbooks regularly contain SEM images of bugs, plants, human tissue, rocks, etc. as a means of illustrating the microscopic world to students. However, despite the widespread use of SEM images for educational purposes microscope companies have virtually ignored the area of education in their development and design of SEMs, due presumably to a perceived lack of marketing potential in this area. This article presents the results of a program aimed at developing a Web-based SEM that is specifically designed for educational use by students and teachers in pre-college classrooms. While virtually all companies tout some form of remote control, no concerted effort has been made to fully develop these capabilities for use by the education sector. The microscope under development, termed the WebSEM, seeks to remedy this situation and address this neglected market. Remote control of the WebSEM is possible by means of a simple web interface that allows the users a wide range of controls, depending on the skill of the operator at the remote site. The web interface is specifically designed to be simple and reliable such that little or no training is required for use. This joint effort between educators and a microscope manufacturer is the first attempt at the development of a truly education-oriented SEM suitable for use in the classroom.

Computer-Assisted Instruction↗
PubMed2001

Human CD38: a (r)evolutionary story of enzymes and receptors.

Human CD38 is the mammalian prototype of a family of proteins which share structural similarities and an ectoenzymatic activity involved in the production of calcium mobilizing compounds. Besides the enzymatic activity, the molecule performs as a receptor, ruling adhesion and signaling in leukocytes. These functions are exerted through the interaction with surface ligands, one of which was identified as CD31. Recently, CD38 has gained attention as a prognostic marker and a pathogenetic agent in leukemias and in other diseases. Together these insights have produced a model of an as yet unique family of molecules, which act independently as receptors and enzymes.

ADP-ribosyl Cyclase↗
PubMed2001

A subtoxic interactive toxicity study of ethanol and chromium in male Wistar rats.

The purpose of this study was to evaluate the interactive toxicity of ethanol with potassium dichromate (K2Cr2O7-chromium). Young, male Wistar rats (100-120 g) were divided into four groups of five or six animals each and were dosed, through water, with 10% ethanol (vol./vol.) or 25 ppm chromium or were dosed with a combination of ethanol+chromium at the same concentrations for a period of 22 weeks ad libitum and were maintained on normal diet. Control animals were maintained on a normal diet and water for the same period. The serum succinate dehydrogenase and liver total triglyceride levels were significantly reduced in the three treated groups. The serum alkaline phosphatase levels were significantly reduced in ethanol-treated rats, and there was no significant change in the acid phosphatase activity. Serum aspartate and alanine aminotransferase levels in the three treated groups were significantly increased. The liver glycogen significantly decreased in both the ethanol-treated and the chromium-treated rats. There was a significant increase in liver total cholesterol levels in chromium-treated rats. Total glutathione levels were significantly decreased in the livers of ethanol-treated and ethanol+chromium-treated rats. To further substantiate these findings, a histological examination of the liver and kidneys was undertaken. The livers of alcohol-treated animals showed altered hepatic architecture in the centrilobular and periportal areas, with increased sinusoidal space (space of Disse), vacuolation, and necrosis of hepatocytes. Similar changes were observed in a histological examination of the livers of chromium-treated rats, except that the damage to the hepatocytes was more confined to the periportal area. Moreover, histological examination of the livers of ethanol+chromium-treated rats revealed uniform damage in the centrilobular and periportal areas, as was observed in the groups treated either with ethanol or chromium. The histological examination of the kidneys in the three treated groups revealed significant damage to the renal tubules and Bowman's capsule, which showed vacuolation and degeneration of the basement membrane. These findings correlate well with the serum enzyme levels found in the treated groups. It is evident from this study that chronic ethanol consumption sensitizes the liver to the toxic action of agents such as chromium. It leads to impairment of the biochemical functions in the liver, and it causes liver and kidney damage. Long-term simultaneous exposure to ethanol and chromium may cause severe health problems in people who are alcoholics and work in chrome-plating and leather-tanning industries.

Alkaline Phosphatase↗
PubMed2001

Mefloquine-induced trigeminal sensory neuropathy.

Trigeminal sensory neuropathy is an important finding, often indicative of trauma but sometimes related to neoplasia, infections, demyelinating conditions, connective tissue disorders, other disorders, or, occasionally, drugs. This paper reports on a patient with sudden-onset trigeminal sensory neuropathy of the lip that proved to be drug-induced, secondary to the antimalarial drug mefloquine. This appears to be the first report of sensory impairment in the orofacial region from exposure to mefloquine.

Antimalarials↗
PubMed2001

Interchain disulfide bonds promote protein cross-linking during protein folding.

We provide evidence that in vitro protein cross-linking can be accomplished in three concerted steps: (i) a change in protein conformation; (ii) formation of interchain disulfide bonds; and (iii) formation of interchain isopeptide cross-links. Oxidative refolding and thermal unfolding of ribonuclease A, lysozyme, and protein disulfide isomerase led to the formation of cross-linked dimers/oligomers as revealed by SDS-polyacrylamide gel electrophoresis. Chemical modification of free amino groups in these proteins or unfolding at pH < 7.0 resulted in a loss of interchain isopeptide cross-linking without affecting interchain disulfide bond cross-linking. Furthermore, preformed interchain disulfide bonds were pivotal for promoting subsequent interchain isopeptide cross-links; no dimers/oligomers were detected when the refolding and unfolding solution contained the reducing agent dithiothreitol. Similarly, the Cys326Ser point mutation in protein disulfide isomerase abrogated its ability to cross-link into homodimers. Heterogeneous proteins become cross-linked following the formation of heteromolecular interchain disulfide bonds during thermal unfolding of a mixture of of ribonuclease A and lysozyme. The absence of glutathione and glutathione disulfide during the unfolding process attenuated both the interchain disulfide bond cross-links and interchain isopeptide cross-links. No dimers/oligomers were detected when the thermal unfolding temperature was lower than the midpoint of thermal denaturation temperature.

Animals↗
PubMed2000

Identification and localization of glutathione S-transferase as a potential target enzyme in Brugia species.

Brugia filarial nematodes are pathogenic lymphatic-dwelling parasites that, like other helminths, may modify the host's defense mechanisms by a major detoxification process involving glutathione-binding proteins such as glutathione S-transferases (GSTs). In the present study, soluble extracts of third-stage larvae, adult male and female worms, microfilariae of either B. pahangi or B. malayi or the adult worm excretory-secretory products of B. malayi were used to determine GST activity. These extracts and affinity-purified fractions of B. pahangi adult worms had a specific enzymatic activity when 1-chloro-2,4-dinitrobenzene was used as a substrate. The observance of this enzyme in all life cycle stages of Brugia spp. demonstrates its ubiquitous nature. Lavage of intraperitoneally infected jirds, but not that of uninfected jirds, also showed increased enzymatic activity, suggesting that GST is secreted in vivo. Soluble proteins of both Brugia spp. were strongly recognized by antibodies in sera from rabbits immunized with affinity-purified native GST of Onchocerca volvulus. Immunohistochemical studies localized these proteins in adult worms, demonstrating cross-reactivity between the GST of these two filarial nematodes. The effect of this enzyme on the motility and viability of adult worms, microfilariae, and larvae was tested in vitro using a battery of known GST inhibitors. Of all those tested, ethacrynic acid, N-ethylmalemide, 4-nitropyridine-oxide, or 1-chloro-2,4-dinitrobenzene at micromolar concentrations reduced the viability and motility of microfilariae, third-stage larvae, and adult worms. These results suggest that Brugia GSTs are major metabolic enzymes and may play an important role in the parasite's survival.

Animals↗
PubMed2000

Trabeculectomy with intraoperative mitomycin C versus 5-fluorouracil. Prospective randomized clinical trial.

OBJECTIVE: To evaluate the relative efficacy and safety of 5-fluorouracil (5-FU) and mitomycin C (MMC) when used as adjuncts with primary trabeculectomy in eyes not at high risk for failure. DESIGN: Prospective multicenter, randomized clinical trial. PARTICIPANTS: One hundred thirteen patients with primary open-angle, pseudoexfoliative, pigmentary, or angle-closure glaucoma undergoing primary trabeculectomy were recruited. METHODS: One eye of each patient was randomized to receive either 5-FU (50 mg/ml for 5 minutes) or MMC (0.4 mg/ml for 2 minutes). MAIN OUTCOME MEASURES: Intraocular pressure (IOP), visual acuity, complications, and interventions were documented at fixed intervals after surgery. The study also examined progression of visual field loss, long-term complications, and bleb appearance 3 years after surgery. RESULTS: Of the 108 patients with complete perioperative information, 54 eyes received 5-FU and 54 received MMC. The proportion of patients reaching different predefined target IOPs after surgery was slightly higher in the MMC group than in the 5-FU group. This difference was less than 25%, which would have been necessary to achieve statistical significance with a power of 0.8 and the sample size used. Likewise, there was no statistically significant difference between the groups with regard to mean preoperative IOP, complications, or interventions. Mean postoperative follow-up was 309 and 330 days in the 5-FU and MMC groups, respectively (P = 0.593). CONCLUSIONS: 5-Fluorouracil and MMC were found to be equally safe and effective adjuncts to primary trabeculectomy in the short- and medium-term postoperative periods.

Aged↗
PubMed2000

Transglutaminase-catalyzed reactions in the growth, maturation and development of parasitic nematodes.

Parasitic nematodes cause several debilitating diseases in humans and animals. New drugs that are parasite specific and minimally toxic to the host are needed to counter these infections effectively. The identification and inhibition of enzymes that are vital for the growth and survival of parasites offer new approaches for developing effective chemotherapeutic agents. Several enzymes in nematodes fall into this category. Here, Ramaswamy Chandrashekar and Kapil Mehta examine in detail the role of transglutaminase, a protein-crosslinking enzyme, in the normal growth and development of nematodes, with an emphasis on filarial parasites.

Amino Acid Sequence↗
PubMed2000

IAEA high-dose intercomparison in 60Co field. International Atomic Energy Agency

Nine high-dose calibration laboratories collaborated with the IAEA in a 'double-blind' intercomparison of absorbed dose in the 10-50 kGy dose range. Bureau International des Poids et Mesure undertook the analysis of the results as well as assisted in the development of the protocol. IAEA alanine-ESR transfer dosimeters were used for this intercomparison. The agreement was within 2.0% at 15 kGy and 2.2% at 45 kGy which is very good considering the variety of irradiation conditions employed by the participants. The mean of the dose values measured by the IAEA Dosimetry Laboratory was within 1% of the mean of the dose values stated by the participants for both dose values.

Journal Article↗
PubMed2000

Temperature stabilization of alanine dosimeters used for food processing and sterilization

The International Atomic Energy Agency has established a dose quality audit service for radiation processing facilities. The objective of the service is to provide an independent check on the routine dosimetry system in use at the facility. The audit service is based on the use of alanine EPR dosimetry. Generally, alanine dosimeters are irradiated at the facility together with a product, and the response is then analyzed at the IAEA laboratory. Practice of the audit service has shown that the main uncertainty in alanine dosimetry is due to absence of temperature control at the irradiation facilities. Here, a method for stabilizing the temperature of the dosimeter during irradiation is proposed.

Journal Article↗
PubMed2000

Downregulation of the p53 tumor suppressor gene and upregulation of the bcl-2 gene in retinoic acid receptor alpha-deficient transgenic mice.

We recently demonstrated lymphoma development in transgenic mice deficient in retinoic acid receptor alpha (RARalpha). High incidence of lymphoma development in this transgenic mouse model system was similar to lymphoma development in p53 knockout mice. In an effort to understand the molecular basis of lymphomagenesis in RARalpha-deficient transgenic mice, we compared the levels of RARalpha to the levels of p53 mRNA, and Bcl-2, and Bax proteins in lymphoid and non-lymphoid tissues and in lymphomas derived from the RARalpha-deficient transgenic mice. The p53 mRNA levels were depleted in various tissues including spleen ( approximately 96%), thymus ( approximately 29%) and bone marrow ( approximately 62%) of RARalpha-deficient transgenic mice when compared with the normal littermates, and the reduction in p53 mRNA expression in the various tissues examined was proportional to the reduction in RARalpha expression. Bcl-2 to Bax ratios were highly increased in the lymphoid compartments (spleen >bone marrow >thymus) because of selective overexpression of Bcl-2 protein. In summary, RARalpha downmodulation in this transgenic mouse model system was accompanied by p53 downmodulation and deregulation of Bcl-2 to Bax ratios in the lymphoid compartments.

Animals↗