PubMed Health⌕ Search

Biomedical subjects

A S Rao

Publications and source records attributed to A S Rao.

At least 19 recordsLinked to original sources
PubMed2005

Wnt-5a has tumor suppressor activity in thyroid carcinoma.

Stabilization of beta-catenin by inhibition of its phosphorylation is characteristic of an activation of the canonical Wnt/beta-catenin signaling pathway and is associated with various human carcinomas. It contrasts to an as yet incompletely characterized action of an alternative noncanonical Wnt signaling pathway on neoplastic transformation. The aim of the present study was to test the effects of a member of the noncanonical Wnt signaling pathway, Wnt-5a, in primary thyroid carcinomas and in thyroid carcinoma cell lines. Compared to normal tissue Wnt-5a mRNA expression was clearly increased in thyroid carcinomas. Immunohistochemically, a bell-shaped response was observed with low to undetectable levels in normal tissue and in anaplastic tumors whereas differentiated thyroid carcinomas showed strong positive immunostaining for Wnt-5a. Transfection of Wnt-5a in a thyroid tumor cell line FTC-133 was able to reduce proliferation, migration, invasiveness and clonogenicity in these cells. These effects of Wnt-5a are associated with membranous beta-catenin translocation and c-myc oncogene suppression and are mediated through an increase in intracellular Ca(2+) release, which via CaMKII pathways promotes beta-catenin phosphorylation. Specific inhibition of beta-catenin phosphorylation by W-7, a calmodulin inhibitor, or by KN-93, a CaMKII inhibitor, supports these findings whereas PKC inhibitors were without effect. This interaction occurs downstream of GSK-3 beta as no Wnt-5a effect was seen on the Ser(9) phosphorylation of GSK-3 beta. Our data are compatible with the hypothesis that Wnt-5a serves as an antagonist to the canonical Wnt-signaling pathway with tumor suppressor activity in differentiated thyroid carcinomas.

Cell Division↗
PubMed2005

Lithium stimulates proliferation in cultured thyrocytes by activating Wnt/beta-catenin signalling.

BACKGROUND: Lithium, clinically used in the treatment of bipolar disorders, is well known to induce thyroid growth. However, the mechanism involved is only incompletely characterized. Although it is conventionally believed that thyroid proliferation depends on the thyroid-stimulating hormone (TSH)/cAMP/cAMP response element binding protein (CREB) pathway, recent data indicate that Wnt/beta-catenin signalling may be of critical importance. In other cell types lithium activates canonical Wnt signalling by GSK-3beta inhibition, which in turn stabilizes cytosolic free beta-catenin. Here we investigated the potential modulation of Wnt/beta-catenin signalling under lithium treatment in primary and neoplastic human thyrocytes. METHODS: Primary (S18) and neoplastic (NPA, FTC133) thyrocytes treated with and without LiCl were analysed using Western blotting, immunoprecipitation, reporter-gene assay, MTT proliferation assay and transfection studies. RESULTS: LiCl dose-dependently inhibited GSK-3beta, stabilized free beta-catenin and inhibited beta-catenin degradation. Furthermore, LiCl altered the assembly of adherens junction by upregulating the E-cad-herin repressor, Snail, and downregulated E-cadherin expression. At a dose of 5 mM, LiCl significantly increased the proliferative potency of thyrocytes, which appeared to be mediated by beta-catenin, since nuclear beta-catenin stimulated T-cell factor/lymphoid enhancer factor (TCF/LEF)-mediated transcription and upregulated downstream targets like cyclin D1. To characterize the specificity of Wnt/beta-catenin-driven thyrocyte proliferation, we transfected primary thyrocytes and FTC133 cells with dominant negative TCF4 to block Wnt-dependent pathways or with dominant negative CREB to inhibit the TSH/cAMP cascade. In cells transfected with dominant negative CREB lithium-stimulated proliferation was unchanged whereas blocking Wnt/beta-catenin by dominant negative TCF4 reduced proliferation by approx. 50%. CONCLUSION: Our data indicate that Wnt/beta-catenin signalling is of major importance in the control of lithium-dependent thyrocyte proliferation.

Blotting, Western↗
PubMed2003

Purification, crystallization and preliminary X-ray diffraction studies of recombinant class A non-specific acid phosphatase of Salmonella typhimurium.

The phoN gene of Salmonella enterica sv. Typhimurium strain MD6001 was cloned in the multicopy plasmid pBluescript SK(-). The nucleotide sequence of the cloned gene differs from the corresponding S. typhimurium LT2 sequence at 23 residues, leading to 15 amino-acid differences, but was very close to the S. typhi phoN sequence (only three nucleotide and two amino-acid differences). The recombinant PhoN protein was purified to homogeneity. Two forms of crystals were harvested from a single crystallization condition. Diffraction intensity data were collected using a laboratory X-ray source to resolution limits of 2.5 and 2.8 A for crystals belonging to space group C2 and C222(1), respectively. Based on non-crystallographic symmetry, four monomers of PhoN are expected to be present in the asymmetric unit of the C2 unit cell. Two monomers of a biologically active dimer in the asymmetric unit of the C222(1) unit cell are expected from the Matthews coefficient.

Acid Phosphatase↗
PubMed2003

Audit on tracheostomies performed at the General Intensive Care Unit, Kuala Lumpur Hospital.

During a 6-month period from October 2000 to March 2001, we analysed the indications, methods, waiting period and complications following a tracheostomy at the General Intensive Care Unit (GIGU) of Hospital Kuala Lumpur. There were 49 tracheostomies performed during this period. Thirty of them were performed in the GICU using the percutaneous dilatational method while 19 were performed electively in the Operating Theatre (OT) by the ear, nose and throat (ENT) surgeons. The main indications for a tracheostomy were prolonged mechanical ventilation and airway protection for patients with a poor Glasgow Coma Scale. The average waiting time for a tracheostomy after a decision was made to perform one was 1.34 +/- 0.72 days for a percutaneous tracheostomy and 3.72 +/- 2.52 days for a surgical tracheostomy. This difference was statistically significant (p < 0.001). There was excessive bleeding in 3 patients in the percutaneous tracheostomy group and 1 patient in the surgical tracheostomy group. Percutaneous tracheostomy is now the main method of tracheostomy at the GICU in Hospital Kuala Lumpur. Haemorrhage is the most significant complication of this procedure. However the overall complication rate is comparable with that of a surgical tracheostomy.

Adult↗
PubMed2003

Optimal size selection of laryngeal mask airway in Malaysian female adult population.

BACKGROUND: The summary of various studies done looking at size selection of the laryngeal mask airway (LMA) in adults is that, selection based on sex is appropriate, and that both sizes 4 or 5 are adequate for adult females. However, in our local population these sizes may be too large especially the size 5 for adult females. OBJECTIVE: To determine the optimal size of LMA in Malaysian female adults. METHOD: 135 ASA 1 or 2 adult female patients coming for elective surgery, requiring general anaesthesia suitable for LMA insertion were randomised into 3 groups to receive either a size 3, 4 or 5 LMA. Optimal size of the LMA was assessed based on 4 parameters, the number of attempts at placement, the oropharyngeal leak pressure (OLP), fibre optic score and the percentage of the vocal cords seen. RESULTS: The 3 groups were demographically similar. There was no difference in the 3 groups in terms of number of attempts of placement, OLP and fibre-optic score. The percentage of vocal cords seen with the size 3 LMA was significantly less than for the size 4 and size 5 (p = 0.009). For the size 5 LMA group in 10/45 patients, the size 5 LMA was too big making it incorrectly positioned after successful insertion and in another 3/45 patients it was difficult to pass the size 5 LMA past the open mouth during insertion. There were no such problems with the size 3 or 4 LMA groups. CONCLUSION: The optimal size of LMA for the female Malaysian adult is size 4.

Adult↗
PubMed2002

Clinical and laboratory evaluation of the safety of a bioartificial liver assist device for potential transmission of porcine endogenous retrovirus.

BACKGROUND: The potential risk of transmission of porcine endogenous retroviruses (PERV) from xenogeneic donors into humans has been widely debated. Because we were involved in a phase I/II clinical trial using a bioartificial liver support system (BLSS), we proceeded to evaluate the biosafety of this device. MATERIALS AND METHODS: The system being evaluated contains primary porcine hepatocytes freshly isolated from pathogen-free, purpose-raised herd. Isolated hepatocytes were installed in the shell, which is separated by a semipermeable membrane (100-kD nominal cutoff) from the lumen through which the patients' whole blood is circulated. Both before and at defined intervals posthemoperfusion, patients' blood was obtained for screening. Additionally, effluent collected from a clinical bioreactor was analyzed. The presence of viral particles was estimated by reverse transcriptase-polymerase chain reaction (RT-PCR) and RT assays. For the detection of pig genomic and mitochondrial DNA, sequence-specific PCR (SS-PCR) was used. Finally, the presence of infectious viral particles in the samples was ascertained by exposure to the PERV-susceptible human cell line HEK-293. RESULTS: PERV transcripts, RT activity, and infectious PERV particles were not detected in the luminal effluent of a bioreactor. Culture supernatant from untreated control or mitogen-treated porcine hepatocytes (cleared of cellular debris) also failed to infect HEK-293 cell lines. Finally, RT-PCR, SS-PCR, and PERV-specific RT assay detected no PERV infection in the blood samples obtained from five study patients both before and at various times post-hemoperfusion. CONCLUSION: Although longer patient follow-up is required and mandated to unequivocally establish the biosafety of this device and related bioartificial organ systems, these analyses support the conclusion that when used under standard operational conditions, the BLSS is safe.

Animals↗
PubMed2002

Purification and characterization of a thermophilic and acidophilic chitinase from Microbispora sp. V2.

AIM: Purification and characterization of a chitinase from Microbispora sp. V2. METHODS AND RESULTS: The chitinase from Microbispora sp. V2 was purified to homogeneity by gel filtration chromatography with 4.6% recovery. It had a molecular weight of 35 kDa and showed maximum activity towards p-nitrophenyl-beta-d-N,N'-diacetylchitobiose, indicating a chitobiosidase activity. The enzyme had a pH optimum of 3.0 and temperature optimum of 60 degrees C. It was stable in a wide pH range from 3.0 to 11.0, retaining 61% activity at pH 3.0 and 52% activity at pH 11.0. It retained 71% activity at 30 degrees C and 45% activity at 50 degrees C, up to 24 h. The enzyme activity was not inhibited by any of the metal ions tested except Hg2+, in the presence of which only 10% activity was retained. CONCLUSIONS: The 35 kDa chitinase from Microbispora sp. V2 has an acidic pH optimum and a high temperature optimum. It is fairly stable and active, and degrades chitin efficiently, although the growth of the culture and enzyme production is slow. SIGNIFICANCE AND IMPACT OF THE STUDY: This report is the first detailed study of a chitinase from Microbispora sp. V2, isolated from hot springs. The chitinase from Microbispora sp. V2 may have potential applications in the recycling of chitinous wastes, particularly due to its thermophilic and acidophilic character. Studies at molecular level may provide further insight on the chitinolytic system of Microbispora spp. with respect to the number and types of chitinases and their regulation.

Actinobacteria↗
PubMed2001

Identification of the CD32/FcgammaRIIc-Q13/STP13 polymorphism using an allele-specific restriction enzyme digestion assay.

We have recently reported that in addition to FcgammaRIIIa (CD16), approximately 45% of normal individuals also express FcgammaRIIc (CD32) on their natural killer (NK) cells. We found this expression to be regulated by an allelic polymorphism localized in the first extracellular exon (EC1) of the FcgammaRIIC gene, corresponding to aa 13. This is determined by a single nucleotide substitution, which results in either a functional open reading frame (glutamine-Q) or a premature stop codon (STP). Identification of this polymorphism provided a good explanation for the lack of CD32 expression previously observed with NK cells in some normal individuals. Here, we describe a new method for detection of FcgammaRIIc allelism based on RT-PCR amplification followed by an allele-specific restriction enzyme digestion. This method is rapid, reliable and time saving, as compared to the currently available allele-specific oligo-nucleotide probe-based Southern Blotting.

Alleles↗
PubMed2001

Interleukin-17 antagonism inhibits acute but not chronic vascular rejection.

BACKGROUND: Blocking the action of interleukin (IL) 17 with an IL-17 receptor (R):Fc fusion protein inhibits T-cell proliferative responses to alloantigens and prolongs vascularized heart graft survival. In this study, we examined whether IL-17 antagonism could suppress the development of chronic rejection. METHODS: A 0.6-cm section of C57BL10 (H2b) thoracic aorta was transplanted to recipient C3H (H2k) abdominal aorta. IL-17R:Fc or control human immunoglobulin G was administered i.p. (500 microg/day) from days 0 to 6 or from days 0 to 29. Mice were killed on days 7 or 30. Grafts were examined histologically and stained for alpha-smooth muscle actin (alpha-smA). Antidonor mixed leukocyte reaction, cytotoxic T cell, and alloantibody responses were quantified. RESULTS: On day 7, control grafts showed mononuclear cell (MNC) infiltration, pronounced endothelial damage, and apoptosis of intimal and medial cell compartments. By day 30, there was concentric intimal thickening, accumulation of alpha-smA+ cells, and collagen deposition. Patchy destruction of the elastic membranes and loss of alpha-smA expression in media were evident. IL-17R:Fc for 6 days decreased MNC infiltration in the intimal and medial compartments at day 7. The endothelium was preserved (completely or partially) in all grafts. The medial compartment showed normal alpha-smA expression. Irrespective of IL-17R:Fc treatment for either 6 days or continuously, allografts harvested at day 30 showed circumferential intimal thickening, with accumulation of alpha-smA+ cells and collagen deposition. There was no effect on circulating alloantibody levels. CONCLUSIONS: These findings support a role for IL-17 in the immunopathogenesis of acute vascular rejection and demonstrate the potential of IL-17 antagonism for therapy. By contrast, IL-17 antagonism does not appear to prevent ensuing chronic graft vascular disease, in particular neointimal formation.

Acute Disease↗
PubMed2001

The concentration of pesticides in sediments from Kolleru Lake in India.

Kolleru Lake is the largest natural freshwater lake in India. Sediment samples were collected at different locations from Kolleru Lake at different seasons in a year over a period of three years and analyzed for eight selected pesticides. In general, pesticides were present at all locations. The selected pesticides alpha-BHC, gamma-BHC, malathion, chlorpyrifos and endosulfan were present at higher concentrations than isodrin, dieldrin and p,p'-DDT. The concentrations of alpha-BHC, gamma-BHC malathion, chlorpyrifos, isodrin, endosulfan, dieldrin and p,p'-DDT in sediments were 1.2-388, 1.4-212, from below the minimum detection limit (BDL)-186, 1.1-292, BDL-38.6, BDL-206, BDL-128 and BDL-191 micrograms kg-1 dry weight, respectively. The aim of this study is to evaluate baseline data, to try to account for the concentration of pesticides in sediments and to discuss their potential to affect human beings and aquatic organisms.

Ecosystem↗
PubMed2001

Crop residue addition effects on myriad forms and sorption of phosphorus in a Vertisol.

Crop residues are a vital organic resource and their extensive use in soil management for sustainable agriculture is widely advocated. The effects of soybean residue (SR) and wheat residue (WR) applied alone or in combination with fertilizer P (FP) on dynamics of labile P, distribution of P fractions and P sorption in a Vertisol (Typic Haplustert) were assessed in a 16 week long incubation study. The amount of P added through crop residues, FP or their combination was kept constant at 10 mg P kg(-1) soil. Addition of SR or WR resulted in net increase of labile inorganic (Pi) and organic (Po) P, and microbial P throughout the incubation period, except that the WR decreased labile Pi during the first two weeks due to Pi immobilization. Integration of FP with SR had no added benefit compared to SR alone, while use of FP + WR proved better in ensuring short-term P availability by offsetting initial P immobilization associated with WR alone. Sequential fractionation of soil P at the end of 16 weeks showed that addition of SR and WR alone or in combination with FP favoured a build-up in labile Pi and Po (NaHCO3-Pi and Po), and moderately labile Po (NaOH-Po) fractions at the expense of recalcitrant P (HCl-P). The P sorption capacity of soil and P required to maintain optimum solution P concentration of 0.2 mg P 1(-1) also decreased with addition of these crop residues. The implication of the results of this study is that soybean and wheat residues can potentially improve soil P fertility by increasing labile Pi and Po, and moderately labile Po fractions, decreasing P sorption and concomitantly causing dissolution of recalcitrant P in soil.

Crops, Agricultural↗
PubMed2001

Novel method for isolation of major phenolic constituents from cashew (Anacardium occidentale L.) nut shell liquid.

Commercially available cashew (Anacardium occidentale L.) nut shell liquid (CNSL) mainly contains the phenolic constituents anacardic acid, cardol, and cardanol. These phenolic constituents are themselves heterogeneous, and each of them contains saturated, monoene, diene, and trienes in the fifteen-carbon side chain. This communication describes the separation of anacardic acid, cardol, and cardanol for industrial application. Anacardic acid was selectively isolated as calcium anacardate. The acid-free CNSL was treated with liquor ammonia and extracted with hexane/ethyl acetate (98:2) to separate the mono phenolic component, cardanol. Subsequently, ammonia solution was extracted with ethyl acetate/hexane (80:20) to obtain cardol.

Chromatography, High Pressure Liquid↗
PubMed2001

Quantitation of immunosuppression by tacrolimus using flow cytometric analysis of interleukin-2 and interferon-gamma inhibition in CD8(-) and CD8(+) peripheral blood T cells.

The authors have determined the frequency of intracellular interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) synthesis by T-cell subsets in whole blood (WB) and isolated lymphocytes in 16 transplant recipients treated with tacrolimus and 10 control patients who were not transplant recipients. The authors also determined the impact of varying amounts of red blood cells (RBC) on immunosuppression by tacrolimus. Samples were analyzed by two-color flow cytometry, and the results were expressed as a ratio of whole blood to isolated lymphocytes. In healthy subjects who were not transplant recipients, the frequency of IL-2--producing CD8(-) and CD8(+) cells was higher in WB than in isolated lymphocytes (mean +/- SD of whole blood to lymphocytes ratio: 1.24 +/- 0.5 and 1.67 +/- 0.62, respectively). Adding varying amounts of RBC had no significant impact on IL-2 production by CD8(-) and CD8(+) T cells. Adding tacrolimus (10 ng/mL) to lymphocyte cultures inhibited (90%) IL-2 production in isolated T cells but not in the whole-blood assay. The dose of tacrolimus required for a 50% inhibition of IL-2 release in T cells was 10-fold higher in cultures with RBC than without. Peripheral blood mononuclear cells (PBMC) isolated from tacrolimus-treated whole blood (WB) showed less IL-2 inhibition than did lymphocytes in the WB. The authors also tested cytokine production in WB and PBMCs in 16 transplant recipients and observed various patterns of reactivity. The frequency of IL-2--producing CD8(-) and CD8(+) cells was similar using two different methods in 10 of 16 patients tested. By contrast, in the remaining six patients the authors observed a significant inhibition of IL-2 production in both CD8(-) and CD8(+) T-cell subsets in the whole-blood assay but not in the isolated lymphocytes. The frequency of CD8(-) IFN-gamma--producing cells was significantly lower in 9 of 16 patients, but the same individuals showed no inhibition of their CD8(+) IFN-gamma T cells. The trough levels of tacrolimus did not predict the level of cytokine inhibition in the whole-blood assay in these patients. The authors' results show that the whole-blood assay for cytokine production can be used for monitoring the in vivo effect of tacrolimus in transplant recipients.

Adult↗
PubMed2001

Quality of drinking water from ponds in villages of Kolleru Lake region.

Kolleru Lake is the largest natural freshwater lake in the districts of East and West Godavari of Andhra Pradesh. The major population centres in the Kolleru Lake region are the 148 villages of which 50 bed villages and 98 belt villages. All bed and belt villages in lake region have at least one drinking water pond. Drinking water ponds are filled with lake water during monsoon season and directly supplied to the public throughout the year. The water samples were collected from village drinking water ponds in a year by covering three seasons and analysed for different physico-chemical parameters to assess the quality of drinking water.

Environmental Monitoring↗
PubMed2001

Heavy metals concentrations in the sediments from Kolleru Lake, India.

Kolleru Lake is the largest natural freshwater lake of Andhra Pradesh in India. It is situated between the latitudes 16 degrees 32(1) and 16 degrees 47(1)N and longitude 81 degrees 05(1) and 81 degrees 21(1) E. The catchment area of the lake is 4763 sq. km. The sediment samples were collected at different points from Kolleru Lake in three seasons a year over a period of three years and analyzed for heavy metals and organic matter. Concentrations of Cu, Pd, Cd, Mn, Ni, Co, Fe and Zn in the sediments were analysed by using Atomic absorption spectrometer (AAS) and selected sediments samples were analysed for Be, Sr, Ba, B, Mo, Tl, V, Cr, Ag, Bi, As and Se by using inductively coupled plasma Mass Spectrometer (ICP-MS).

Environmental Monitoring↗