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Biomedical subjects

S Kumar

Publications and source records attributed to S Kumar.

At least 55 records · Page 3Linked to original sources
PubMed2006

Development and characterization of a highly specific and sensitive sandwich ELISA for detection of aggrecanase-generated aggrecan fragments.

OBJECTIVE: To develop an enzyme linked immunosorbent assay (ELISA) to quantify the levels of specific aggrecan fragments generated by aggrecanase-mediated cleavage at the 373Glu-374 Ala bond within the aggrecan interglobular domain. METHODS: The ELISA employs a commercially available monoclonal antibody to capture aggrecan fragments containing keratan sulfate (KS). Aggrecan fragments generated by cleavage at the Glu-Ala bond were then detected using a monoclonal neoepitope antibody (mAb OA-1) that specifically recognizes the N-terminal sequence 'ARGSVIL'. RESULTS: The mAb OA-1 antibody was highly specific for the immunizing neoepitope peptide since neither peptides spanning the cleavage site nor mutated peptides were detected. Aggrecan fragments generated by ADAMTS-4 digested human aggrecan monomers and from IL-1-stimulated human cartilage explants were quantified by the ELISA, and we observed increased sensitivity of the ELISA compared to mAb OA-1 Western analysis. We also observed that the basal, as well as IL-1-stimulated production of ARGS aggrecan fragments from human articular cartilage explants was blocked by a selective aggrecanase inhibitor, consistent with generation of the ARGS neoepitope in human articular cartilage being mediated by aggrecanase. Using purified human aggrecan digested by ADAMTS-4 as standard to quantify ARGS aggrecan fragments in human synovial fluids, we determined that the calculated amount of ARGSVIL-aggrecan fragments by ELISA measurement is in agreement with the published levels of these fragments, supporting its potential utility as a biomarker assay for osteoarthritis. CONCLUSION: We have developed an assay that detects and quantifies specific aggrecan fragments generated by aggrecanase-mediated cleavage. Because aggrecanase mediates degradation of human articular aggrecan in joint disease, the KS/mAb OA-1 ELISA may serve as a biomarker assay for evaluation of preclinical and clinical samples.

ADAM Proteins↗
PubMed2006

Ultrasound biomicroscopic quantification of the change in anterior chamber angle following laser peripheral iridotomy in early chronic primary angle closure glaucoma.

AIMS: To prospectively evaluate by ultrasound biomicroscopy (UBM) and gonioscopy the anterior chamber angle widening following laser peripheral iridotomy (LPI) in eyes with early chronic primary angle closure glaucoma (CACG). METHODS: A total of 55 eyes of 55 patients with CACG presenting with less than 180 degrees peripheral anterior synechiae (PAS) were enrolled in the study. Angles were assessed by gonioscopy (Shaffer's grading) and UBM, before and 4 weeks after LPI. The angle opening distance at 250 and 500 microm from the scleral spur (AOD 250 and AOD 500) was computed. Results were analysed using the Wilcoxon signed-rank test. RESULTS: In the quadrant with LPI, the mean gonioscopy grade increased significantly from 0.45 to 1.45 (P<0.001) and the mean AOD 250 and AOD 500 increased from 38.5+/-25.9 to 83.5+/-48.4 microm (P<0.001) and 110.2+/-80.9 to 170.6+/-83.4 microm (P<0.001), respectively. The angles widened significantly in the opposite quadrant on UBM (AOD 250: 48.8+/-31.5-82.7+/-43.9 microm, P<0.001; AOD 500:117.2+/-65.5-172.2+/-81.7 microm; P<0.001), but the median gonioscopy grade remained unchanged. CONCLUSIONS: LPI significantly widened the anterior chamber angle in the quadrant with LPI and the quadrant furthest away in patients of CACG with established glaucomatous damage. This change was much better appreciated by the UBM than gonioscopy.

Adult↗
PubMed2006

Fentanyl-augmented MRCP.

BACKGROUND: Drugs such as secretin and morphine have been used to augment the visualization of magnetic resonance cholangiopancreatography (MRCP). This study investigated the effectiveness of intravenous administration of a synthetic opioid, fentanyl, in improving the MRCP image quality. METHODS: Thirty consecutive patients with a provisional diagnosis of benign biliary and/or pancreatic disease underwent MRCP. Coronal single-shot fast spin-echo heavily T2-weighted dynamic MRCP images were generated before and at every minute for 10 min after intravenous administration of fentanyl citrate at a dose of 1.0 mug/kg. Pre- and postinjection images were compared and analyzed qualitatively and quantitatively. RESULTS: Qualitatively, visualization of intrahepatic bile ducts, common bile duct, and main pancreatic duct improved after fentanyl injection in five (16%), 11 (37%), and 19 (63%) patients, respectively. The pancreatobiliary junction and common channel were visualized better after fentanyl injection in eight of the 18 patients (44%). Quantitatively, signal intensity and diameters of the intrahepatic ducts, common bile duct, and main pancreatic duct measured at corresponding points on pre- and postinjection images showed an increase above preinjection values in 28 (93%), 27 (90%), and 21 (70%) and in 18 (60%), 26 (86%), and 22 (73%), respectively, and these changes were highly significant at all sites (p < 0.001). CONCLUSIONS: Intravenous administration of fentanyl before MRCP improves qualitative and quantitative visualization of the ductal system anatomy that may be of value in clinical diagnosis and management.

Adolescent↗
PubMed2006

Functional characterization of an orphan nuclear receptor, Rev-ErbAalpha, in chondrocytes and its potential role in osteoarthritis.

OBJECTIVE: To evaluate the expression and function of the orphan nuclear receptor Rev-ErbAalpha in articular cartilage and to investigate its role in osteoarthritis (OA). METHODS: Expression of Rev-ErbAalpha was analyzed at both the messenger RNA and protein levels in human and bovine articular cartilage and chondrocytes by real-time polymerase chain reaction (TaqMan) and immunocytochemical techniques. The effects of cartilage catabolic and anabolic agents on the expression of Rev-ErbAalpha were evaluated by TaqMan analysis. Overexpression was achieved by either adenoviral transduction or treatment with a peroxisome proliferator-activated receptor alpha agonist, whereas expression was suppressed by antisense oligonucleotides. RESULTS: Among the 48 known nuclear receptors, Rev-ErbAalpha was found to be the most highly expressed in OA cartilage. It is known to function as a transcription repressor. Treatment of articular chondrocytes with known catabolic agents resulted in the induction of Rev-ErbAalpha, whereas stimulation with anabolic agents led to a decrease in expression. Overexpression of the nuclear receptor was associated with an increase in the expression of matrix-degrading enzymes such as matrix metalloproteinase 13 and aggrecanase. In contrast, a decrease in Rev-ErbAalpha expression led to a concomitant reduction in the activity of matrix-degrading enzymes. CONCLUSION: This study is the first to demonstrate that Rev-ErbAalpha is highly expressed in OA articular chondrocytes and that its expression is modulated by known cartilage catabolic and anabolic stimuli. We also demonstrated that modulation of Rev-ErbAalpha expression in chondrocytes may be a novel means of regulating the expression and production of multiple matrix-degrading enzymes. These observations suggest that Rev-ErbAalpha may be a novel therapeutic target for OA.

ADAM Proteins↗
PubMed2006

Outcome in the real-world of coronary high-risk intervention with drug-eluting stents (ORCHID) -- a single-center study comparing Cypher sirolimus-eluting with Taxus paclitaxel-eluting stents.

OBJECTIVE: We present real world experience from a single center registry comparing the 6-month outcome of percutaneous coronary intervention (PCI) in unselected high-risk individuals using either sirolimus-eluting (SES) or paclitaxel-eluting stents (PES). METHODS/RESULTS: We compared clinical outcome at 6 months follow-up in two cohorts of 156 consecutive patients (total n = 312) who underwent SES (June 2002-February 2003) and PES (march 2003-July 2003) implantation. The primary endpoint was a composite of major adverse cardiac events (MACE). Baseline clinical characteristics were well matched. The 6-month target vessel revascularization (TVR) rates were 1.9% (SES) and 2.6% (PES) and MACE rates were similar in the two groups (SES 4.5% vs. PES 3.2%, P = NS). In the PES group, intervention for multivessel disease, bifurcation lesions and in small vessels was more common, and for in-stent restenosis less common, reflecting the impact of drug eluting stents on indications for PCI. The incidence of sub-acute stent thrombosis, related to inadequate antiplatelet therapy in 3 of the 6 cases, was 0.95% with no difference between the two groups. CONCLUSION: This study confirms the safety and efficacy of SES and PES in unselected high risk patients undergoing PCI. Clinical outcomes of both stents are equivalent at 6 months with low rates of MACE and TVR. These data provide important complementary information to forthcoming randomized studies.

Angioplasty, Balloon, Coronary↗
PubMed2006

Day case transradial coronary angioplasty: a four-year single-center experience.

We examined the safety and feasibility of elective outpatient transradial coronary angioplasty (PCI). Four hundred and forty two patients underwent procedures over a 4-year period. Over 95% had an excellent angiographic result and 85% were discharged the same day. Radial access was successful in 417 (94%) patients. There were no major vascular complications. One patient died of a subacute stent thrombosis. Outpatient transradial PCI is safe and feasible for the majority of elective PCI cases.

Aged↗
PubMed2006

Enhanced transduction of mouse bone marrow-derived dendritic cells by repetitive infection with self-complementary adeno-associated virus 6 combined with immunostimulatory ligands.

The potential of adeno-associated virus (AAV)-based vectors in human gene therapy is being explored for several diseases. Although sustained transgene expression and low vector-associated cellular immunity are attractive features of recombinant (r) AAV, the wider application of rAAV vectors encapsidated in serotype 2 capsid is hampered by poor transduction efficiency in many target tissues. These include ex vivo-generated dendritic cells (DC), which have demonstrated promising immunotherapeutic activity. We report here that efficient transduction of mouse bone marrow-derived DC can be achieved with self-complementary (sc) rAAV encapsidated in serotype 6 capsid. Sequential exposure of DC precursor cultures to IL-4 and GM-CSF with sc rAAV6 encoding the human tumor antigen, carcinoembryonic antigen (CEA), for 7 days followed by activation with CpG oligodeoxynucleotides (ODN) and anti-mouse CD40 antibody resulted in highly efficient transduction of DC. DC surface markers as determined by flow cytometry analysis of sc rAAV6-transduced DC were comparable to nontransduced DC. Efficiency of vector transduction and transgene expression were confirmed by immunostaining and real-time PCR. Microarray analysis of RNA from CpG ODN and CD40 antibody stimulated sc AAV6-transduced DC revealed upregulation of transcription factors and cytokines involved in immune activation and downregulation of inhibitory factors, suggesting a possible role of transcriptional activation in the observed effect. The adoptive transfer into syngeneic mice of the ex vivo-transduced and activated DC resulted in the development of CEA-specific antibody and T-helper 1-associated immune responses. Immunized mice also developed antibody to AAV6 capsid protein, which did not crossreact with AAV2 capsid protein. These studies demonstrate the potential utility of sc rAAV serotype 6-based vectors in transduction of DC for genetic vaccination approaches.

Adoptive Transfer↗
PubMed2006

Prostate atrophy and spinal cord lesions.

STUDY DESIGN: A single blinded, quantifiable survey of prostate size in spinal cord injury (SCI) patients. OBJECTIVE: A small prostate gland is sometimes found on routine digital rectal examination (DRE) in SCI patients. Interruption of neurohormonal supply to the prostate gland might lead to atrophy. To test this interpretation, transrectal ultrasonic (TRUS) examinations have been carried out in SCI patients stratified by severity of paralysis. SUBJECTS: Nine severely paralyzed spinal cord-injured men (levels at T10 or above and ASIA A, B, or C) were compared with 12 less severely paralyzed men (levels lower than T10 at any grade or ASIA D at any level). The groups were age matched. METHODS: All patients were examined with a 90 degrees sector TRUS probe that measured the prostate gland in three dimensions and calculated the prostate volume and weight. Prostate-specific antigen (PSA) levels were also measured. RESULTS: By TRUS the prostate size was smaller in the severely paralyzed (range 8-16 g, mean 13 g) than in the less severely paralyzed (range 10-70 g, mean 28 g), P=0.02. The PSA level of the severely paralyzed group tended to be lower (0.5-2.1, mean 0.7 ng/ml versus 0.5-10.4, mean 2.2 ng/ml), P=0.08. CONCLUSION: The prostate gland of severely paralyzed SCI patients is small. Interruption of neurohormonal pathways due to extensive cord damage may be a factor.

Aged↗
PubMed2006

Detection of Salmonella enterica serovar Typhi (S. Typhi) by selective amplification of invA, viaB, fliC-d and prt genes by polymerase chain reaction in mutiplex format.

AIMS: Development of a PCR assay that can target multiple genes for rapid detection of Salmonella enterica serovar Typhi (S. Typhi) from water and food samples. METHODS AND RESULTS: PCR primers for invasion, O, H and Vi antigen genes, invA, prt, fliC-d and viaB were designed and used for the rapid detection of S. Typhi by multiplex PCR. Internal amplification control, which co-amplified with prt primers, was also included in the assay. The results showed that all cultures of Salmonella were accurately identified by the assay with no nonspecific amplification in other cultures. The assay had 100% detection probability when a cell suspension of 10(4) CFU ml(-1) (500 CFU per reaction) was used. Salmonella Typhi bacteria were artificially inoculated in the water and food (milk and meat rinse) samples and detected by mPCR after overnight pre-enrichment in buffered peptone water. No Salmonella bacteria could be detected from water samples collected from the field by mPCR or standard culture method. CONCLUSIONS: The developed mPCR assay provides specific detection of S. Typhi. SIGNIFICANCE AND IMPACT OF THE STUDY: Rapid methods for detection of S. Typhi from complex environmental matrices are almost nonexistent. The mPCR assay reported in this study can be useful to identify S. Typhi bacteria in field environmental samples.

Antigens, Bacterial↗
PubMed2006

An antiviral protein having deoxyribonuclease and ribonuclease activity from leaves of the post-flowering stage of Celosia cristata.

An antiviral protein named CCP-27 was purified from the leaves of Celosia cristata at the post-flowering stage by anion-exchange, cation-exchange, and gel-filtration chromatography. It exhibited resistance against sunnhemp rosette virus in its test host Cyamopsis tetragonoloba. It also exhibited deoxyribonuclease activity against supercoiled pBlueScript SK+ plasmid DNA. It was found to nick supercoiled DNA into nicked circular form at lower protein concentration followed by nicked to linear form conversion at higher protein concentration. CCP-27 also possesses strong ribonuclease activity against Torula yeast rRNA.

Antitussive Agents↗
PubMed2006

Shock wave lithotripsy in solitary functioning kidneys: is prophylactic stenting necessary?

OBJECTIVE: To assess the safety of shock wave lithotripsy (SWL) without prophylactic stents in solitary functioning kidneys. PATIENTS AND METHODS: Sixteen solitary functioning kidneys with 23 renal stones with a size of <15 mm were treated with SWL as the primary modality. All patients were counseled about the possibility of obstruction, and treatment was offered to those who consented. The safety of SWL was assessed by the need for interventions and the posttreatment renal function. RESULTS: In 14 patients lithotripsy was uneventful. The duration of treatment ranged from 5 to 35 days. One patient with a 15-mm pelvic calculus presented with anuria which resolved before intervention. In 1 patient fragmentation failed, and percutaneous nephrolithotomy was performed. CONCLUSIONS: In solitary functioning kidneys, SWL is safe without prophylactic stents in properly selected and closely monitored patients. Avoiding stents decreases costs, duration of treatment, and stent-related morbidity without unduly compromising safety.

Adult↗
PubMed2006

Pathophysiology of acute ischaemic stroke: an analysis of common signalling mechanisms and identification of new molecular targets.

Stroke continues to be a major cause of death and disability. The currently available therapies have proven to be highly unsatisfactory (except thrombolysis) and attempts are being made to identify and characterize signalling proteins which could be exploited to design novel therapeutic modalities. The pathophysiology of stroke is a complex process. Delaying interventions from the first hours to days or even weeks following blood vessel occlusion may lead to worsening or impairment of recovery in later stages. The objective of this review is to critically evaluate the major mechanisms underlying stroke pathophysiology, especially the role of cell signalling in excitotoxicity, inflammation, apoptosis, neuroprotection and angiogenesis, and highlight potential novel targets for drug discovery.

Animals↗
PubMed2006

Effect of time delay on survival in patients with non-small cell lung cancer--a Malaysian study.

The prognosis of lung cancer remains poor with overall five year survival figures varying between five and 10% worldwide, However, it has been shown that surgery in patients with early stage disease in non-small cell lung cancer can achieve five year survival rates up to 80%, suggesting that early or delay diagnosis can influence prognosis. Nevertheless, studies addressing this have been inconclusive and mostly derived from Western countries.

Carcinoma, Non-Small-Cell Lung↗