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Biomedical subjects

K Sharma

Publications and source records attributed to K Sharma.

At least 19 recordsLinked to original sources
PubMed2001

Direct cleavage of the human DNA fragmentation factor-45 by granzyme B induces caspase-activated DNase release and DNA fragmentation.

The protease granzyme B (GrB) plays a key role in the cytocidal activity during cytotoxic T lymphocyte (CTL)-mediated programmed cell death. Multiple caspases have been identified as direct substrates for GrB, suggesting that the activation of caspases constitutes an important event during CTL-induced cell death. However, recent studies have provided evidence for caspase-independent pathway(s) during CTL-mediated apoptosis. In this study, we demonstrate caspase-independent and direct cleavage of the 45 kDa unit of DNA fragmentation factor (DFF45) by GrB both in vitro and in vivo. Using a novel and selective caspase-3 inhibitor, we show the ability of GrB to process DFF45 directly and mediate DNA fragmentation in the absence of caspase-3 activity. Furthermore, studies with DFF45 mutants reveal that both caspase-3 and GrB share a common cleavage site, which is necessary and sufficient to induce DNA fragmentation in target cells during apoptosis. Together, our data suggest that CTLs possess alternative mechanism(s) for inducing DNA fragmentation without the requirement for caspases.

Apoptosis↗
PubMed2001

Neuroprotective effects of Withania somnifera Dunn. in hippocampal sub-regions of female albino rat.

The neuroprotective effects of W. somnifera were studied on stressed adult female Swiss albino rats. Experimental rats were subjected to immobilization stress for 14 h and were treated with a root powder extract of W. somnifera available as Stresscom capsules (Dabur India Ltd). Control rats were maintained in completely, non stressed conditions. Thionin stained serial coronal sections (7 microm) of brain passing through the hippocampal region of stressed rats (E(1) group) demonstrated 85% degenerating cells (dark cells and pyknotic cells) in the CA(2) and CA(3) sub-areas. Treatment with W. somnifera root powder extract significantly reduced (80%) the number of degenerating cells in both the areas. The study thus demonstrates the antistress neuroprotective effects of W. somnifera.

Animals↗
PubMed2001

Nongestational pure ovarian choriocarcinoma with contralateral teratoma.

BACKGROUND: Nongestational pure ovarian choriocarcinoma is a rare malignancy with only 29 cases described to date. CASE: We describe a case associated with mature cystic teratoma of the contralateral ovary. Details of all 30 cases including clinical features, associations, surgicopathological findings, operative procedures, postoperative chemotherapy, and outcome are summarized. CONCLUSION: This rare malignancy responds well to surgery and postoperative chemotherapy including the methotrexate-based regimes.

Adolescent↗
PubMed2001

Parvalbumin and calbindin D-28K immunoreactive neurons in area MT of rhesus monkey.

The chemical characteristics of the neurons of the motion sensitive visual area, area MT, remain to be established. We studied the distribution pattern of two calcium binding proteins, parvalbumin (PV) and calbindin D28K (CB) in this area, using specific monoclonal antibodies and the peroxidase-antiperoxidase (PAP) immunohistochemical technique. Aldehyde fixed 30-micron-thick cryostat sections from area MT of five animals were processed free floating for immunohistochemical staining. Besides studying the morphological characteristics of PV and CB positive neurons, quantitative analysis was carried out to determine their (1) perikaryal area (Pa) and diameter, (2) numerical densities (NV)/mm3 cortical tissue, (3) absolute number (NC) in a column of cortex under 1 mm2 cortical surface along with (4) layerwise absolute number (NL) under 1 mm2 cortical surface and (5) laminar percentage distribution of immunoreactive (IR) neurons. Quantitative analysis was carried out using a Leica QMC 500 image analysis system connected to a DMRE microscope. The results showed that both types of IR neurons were localized to all cortical layers except layer I. The PV +ve neurons were equidistributed between the supra- and infragranular layers, with the highest percentage being present in layer III (45%) followed by layer V (21%). The CB +ve neurons, on the other hand, were predominantly localized in supragranular layers, with the highest percentage being in layer III (54%) and the next highest percentage in layer II (18%). The average Pa and diameter of PV +ve neurons were found to be 96.90 +/- 28.43 micron 2 and 11.01 +/- 1.61 microns respectively. The CB +ve neurons were significantly smaller in size than the PV +ve neurons, with average Pa and diameter of the former being 92.23 +/- 26.18 micron 2 and 10.39 +/- 1.23 microns respectively. The NV for PV and CB +ve neurons showed ranges of 3157-3894 and 2303-2585, with means of 3347 +/- 285 (+/- SD) and 3436 +/- 100 respectively. The values for NC showed ranges of 5230-5444 and 4020-4268 with means of 5378 +/- 85 and 4167 +/- 95 for PV and CB neurons respectively. Variations in size together with the differential distribution of these neurons in the cortical layers may indicate their involvement in different functional circuitaries.

Animals↗
PubMed2001

Development of the limb neuromuscular system.

Appendages, such as wings of a fly or limbs of a vertebrate, are excellent models to study the principles of patterning and morphogenesis. In the adult these structures are used for a variety of behaviors, including locomotion. Although support structures of the adult vertebrate limb are generated within the limb bud, its dynamic elements are derived from the somitic mesoderm and neural tube. Recent studies show that regional patterns set up in the mesenchyme-filled limb bud guide muscle precursors and developing motor axons to their proper location within the limb. Subsequent development of the neuromuscular system is regulated by cell surface interactions between pre-specified muscle fibers and motor axons.

Animals↗
PubMed2001

Effect of anti-transforming growth factor-beta antibodies in cyclosporine-induced renal dysfunction.

BACKGROUND: Several experimental and clinical studies have implicated a role for transforming growth factor-beta (TGF-beta) in mediating the nephrotoxic effects of cyclosporine (CsA). To test this hypothesis, we administered neutralizing anti-TGF-beta antibodies (alpha-TGF-beta) in a well-described rat model of chronic CsA nephrotoxicity. METHODS: We studied three groups (N = 9 per group) of adult, male Sprague-Dawley rats that received a low-salt diet (0.05% sodium). Normal controls were given vehicle subcutaneously and an alternate-day intraperitoneal injection of 3 mg of nonspecific mouse IgG (MIgG) for 28 days. The CsA group received 15 mg/kg/day of CsA subcutaneously and 3 mg of MIgG intraperitoneally on alternate days for 28 days. The CsA/alpha-TGF-beta group received CsA and alternate-day alpha-TGF-beta (3 mg) for 28 days. At the end of 28 days, creatinine clearance was measured by 24-hour urine collection. Histologic assessment was performed for tubulointerstitial damage and arteriolar hyalinosis. Northern analysis was performed for alpha 1(I) collagen and TGF-beta 1 gene expression, and quantitative reverse transcription-polymerase chain reaction was performed to measure levels of tissue inhibitor of metalloproteinase-1 (TIMP-1), TIMP-2, plasminogen activator inhibitor-1 (PAI-1), matrix metalloproteinase-2 (MMP-2), and MMP-9. RESULTS: CsA-treated rats had significantly lower creatinine clearance as compared with normal controls (0.43 +/- 0.07 vs. 0.67 +/- 0.14 mL/min, P = 0.0002), increased interstitial damage and afferent arteriolar hyalinosis (P = 0.0001), and increased alpha1(I) collagen (4-fold) and TGF-beta 1 (2.5-fold) mRNA expression. CsA-treated rats also had significantly increased TIMP-1 (7.4-fold, P < 0.001), MMP-2, and PAI-1 (all approximately 2-fold, P < 0.02) and decreased MMP-9 (85% reduction, P < 0.001) as compared with controls. Treatment with alpha-TGF-beta in CsA-treated rats significantly prevented the reduction in creatinine clearance (0.58 +/- 0.03 mL/min, P = 0.009 vs. CsA alone), the increase in afferent arteriolar hyalinosis (P < 0.05 vs. CsA alone), normalized alpha 1(I) collagen mRNA levels, and attenuated CsA effects on TGF-beta1, TIMP-1, and MMP-9. CONCLUSIONS: In this rat model of CsA-induced nephrotoxicity, renal insufficiency and characteristic histologic changes are associated with altered expression of matrix and matrix-regulating molecules. Based on our results with alpha-TGF-beta antibodies, many but not all of these nephrotoxic effects of CsA are mediated by TGF-beta.

Animals↗
PubMed2001

Roles for internal and flanking sequences in regulating the activity of mating-type-silencer-associated replication origins in Saccharomyces cerevisiae.

ARS301 and ARS302 are inactive replication origins located at the left end of budding yeast (Saccharomyces cerevisiae) chromosome III, where they are associated with the HML-E and -I silencers of the HML mating type cassette. Although they function as replication origins in plasmids, they do not serve as origins in their normal chromosomal locations, because they are programmed to fire so late in S phase that they are passively replicated by the replication fork from neighboring early-firing ARS305 before they have a chance to fire on their own. We asked whether the nucleotide sequences required for plasmid origin function of these silencer-associated chromosomally inactive origins differ from the sequences needed for plasmid origin function by nonsilencer-associated chromosomally active origins. We could not detect consistent differences in sequence requirements for the two types of origins. Next, we asked whether sequences within or flanking these origins are responsible for their chromosomal inactivity. Our results demonstrate that both flanking and internal sequences contribute to chromosomal inactivity, presumably by programming these origins to fire late in S phase. In ARS301, the function of the internal sequences determining chromosomal inactivity is dependent on the checkpoint proteins Mec1p and Rad53p.

Cell Cycle Proteins↗
PubMed2001

Hoxd10 induction and regionalization in the developing lumbosacral spinal cord.

We have used Hoxd10 expression as a primary marker of the lumbosacral region to examine the early programming of regional characteristics within the posterior spinal cord of the chick embryo. Hoxd10 is uniquely expressed at a high level in the lumbosacral cord, from the earliest stages of motor column formation through stages of motoneuron axon outgrowth. To define the time period when this gene pattern is determined, we assessed Hoxd10 expression after transposition of lumbosacral and thoracic segments at early neural tube stages. We present evidence that there is an early prepattern for Hoxd10 expression in the lumbosacral neural tube; a prepattern that is established at or before stages of neural tube closure. Cells within more posterior lumbosacral segments have a greater ability to develop high level Hoxd10 expression than the most anterior lumbosacral segments or thoracic segments. During subsequent neural tube stages, this prepattern is amplified and stabilized by environmental signals such that all lumbosacral segments acquire the ability to develop high levels of Hoxd10, independent of their axial environment. Results from experiments in which posterior neural segments and/or paraxial mesoderm segments were placed at different axial levels suggest that signals setting Hoxd10 expression form a decreasing posterior-to-anterior gradient. Our experiments do not, however, implicate adjacent paraxial mesoderm as the only source of graded signals. We suggest, instead, that signals from more posterior embryonic regions influence Hoxd10 expression after the early establishment of a regional prepattern. Concurrent analyses of patterns of LIM proteins and motor column organization after experimental surgeries suggest that the programming of these characteristics follows similar rules.

Animals↗
PubMed2001

Diabetic nephropathy.

Diabetic nephropathy is the leading cause of end-stage renal disease in the United States and the largest contributor to the total cost of diabetic care. In addition to the development of diabetic nephropathy and end-stage renal failure, diabetic patients with evidence of albuminuria have a much higher risk of developing myocardial infarctions, cerebrovascular accidents, severe progressive retinopathy, and neuropathy. This article characterizes the clinical and pathologic features of diabetic nephropathy and reviews the major pathogenetic theories that underlie the development of this dreaded complication of diabetes. Widespread screening for this condition and aggressive treatment of diabetic nephropathy at early stages of disease are critical to diminish the risk of costly late complications.

Diabetic Nephropathies↗
PubMed2001

Permanent tooth emergence in Gujjars of Punjab, India.

Patterns of permanent tooth emergence in Gujjars were studied in a cross-sectional sample of 483 children ranging in age from 6 to 13 years. Females were markedly advanced in permanent tooth emergence times over males, but no such sex differences were observed in sequence of emergence. Differences between median emergence times of right and left side antimers were significant for only 4 of 28 instances (14.29%), namely central incisors, mandibular first molars in males and lateral maxillary incisors in females. In general mandibular teeth except premolars tended to emerge earlier than their maxillary counterparts. The quiescent period between first and second tooth emergence stages was longer in males than in females. Mandibular depth and morphological facial length were very significantly correlated (p < 0.01) with the number of permanent teeth present in the oral cavity.

Adolescent↗
PubMed2001

Correlation between Congo red binding and contact haemolysin production in Shigella species.

Haemolytic strains of Shigella dysenteriae type 1, Shigella flexneri, Shigella boydii and Shigella sonnei cultured on Congo red agar produced pigmented colonies (Pcr+) whereas nonhaemolytic strains produced white colonies and did not bind Congo red (Pcr-). S. flexneri-1 haemolysin negative mutant (lacking plasmid) of haemolysin positive prototroph also did not bind Congo red and produced nonpigmented colonies. Among the twelve strains of Shigella included in this study, the characteristics of Congo red binding, plasmid profile and haemolytic activity appeared to be correlated. Congo red binding occurred comparatively more by haemolysin-producing strains. Congo red binding can be used as a quick and reliable method for virulence traits of pathogens, including haemolysin activity.

Bacterial Adhesion↗
PubMed2001

Association between ACE gene polymorphism and diabetic nephropathy in South Indian patients.

OBJECTIVE: To study the association of ACE gene polymorphism and diabetic nephropathy in South Indian subjects. SETTING: Outpatient clinic of a specialized hospital. PATIENTS: The study included 109 South Indian type 2 diabetic patients (72 males and 37 females; age 56.7 plus/minus 9.0 years, mean plus/minus SD). The patients were subdivided into two groups: nephropathic (n=86) and normoalbuminuric patients (n=23). INTERVENTIONS: Genomic DNA was isolated from the peripheral blood leukocytes. To determine the ACE genotype, genomic DNA was amplified by PCR initially using a flanking primer pair and, subsequently when necessary, with a primer pair that recognizes the insertion specific sequence for confirmation of the specificity of the amplification reactions. MAIN OUTCOME MEASURES: ACE genotype distribution in the two study groups. RESULTS: In the nephropathic patients, ID and DD genotypes were present in 52.3% and 27.9% of the patients, respectively as compared to 34.8% and 21.7% respectively in those with normoalbuminuria. The D allele was present in 80.2% of the nephropathic patients and 56.5% of the normoalbuminuric patients (chi-squared=4.28, P=0.039; odds ratio 3.12). Therefore, the higher percentage of II genotype in the normoalbuminuric group was 43.5% as compared to the 19.8% in nephropathic patients. CONCLUSIONS: This study showed a positive association between the D allele (ID and DD genotype) of the ACE polymorphism and diabetic proteinuria in South Indian type 2 diabetic patients. Our findings are in keeping with several earlier studies showing a strong association of the D allele of the ACE gene with diabetic nephropathy.

Alleles↗
PubMed2000

Genetic and epigenetic mechanisms contribute to motor neuron pathfinding.

Many lines of evidence indicate that genetically distinct subtypes of motor neurons are specified during development, with each type having characteristic properties of axon guidance and cell-body migration. Motor neuron subtypes express unique combinations of LIM-type homeodomain factors that may act as intrinsic genetic regulators of the cytoskeletal events that mediate cell migration, axon navigation or both. Although experimentally displaced motor neurons can pioneer new routes to their targets, in many cases the axons of motor neurons in complete isolation from their normal territories passively follow stereotypical pathways dictated by the environment. To investigate the nonspecific versus genetically controlled regulation of motor connectivity we forced all motor neurons to express ectopically a LIM gene combination appropriate for the subgroup that innervates axial muscles. Here we show that this genetic alteration is sufficient to convert the cell body settling pattern, gene-expression profile and axonal projections of all motor neurons to that of the axial subclass. Nevertheless, elevated occupancy of the axial pathway can override their genetic program, causing some axons to project to alternative targets.

Animals↗
PubMed2000

Long-term prevention of renal insufficiency, excess matrix gene expression, and glomerular mesangial matrix expansion by treatment with monoclonal antitransforming growth factor-beta antibody in db/db diabetic mice.

Emerging evidence suggests that transforming growth factor-beta (TGF-beta) is an important mediator of diabetic nephropathy. We showed previously that short-term treatment with a neutralizing monoclonal anti-TGF-beta antibody (alphaT) in streptozotocin-diabetic mice prevents early changes of renal hypertrophy and increased matrix mRNA. To establish that overactivity of the renal TGF-beta system mediates the functional and structural changes of the more advanced stages of nephropathy, we tested whether chronic administration of alphaT prevents renal insufficiency and glomerulosclerosis in the db/db mouse, a model of type 2 diabetes that develops overt nephropathy. Diabetic db/db mice and nondiabetic db/m littermates were treated intraperitoneally with alphaT or control IgG, 300 microgram three times per week for 8 wk. Treatment with alphaT, but not with IgG, significantly decreased the plasma TGF-beta1 concentration without decreasing the plasma glucose concentration. The IgG-treated db/db mice developed albuminuria, renal insufficiency, and glomerular mesangial matrix expansion associated with increased renal mRNAs encoding alpha1(IV) collagen and fibronectin. On the other hand, treatment with alphaT completely prevented the increase in plasma creatinine concentration, the decrease in urinary creatinine clearance, and the expansion of mesangial matrix in db/db mice. The increase in renal matrix mRNAs was substantially attenuated, but the excretion of urinary albumin factored for creatinine clearance was not significantly affected by alphaT treatment. We conclude that chronic inhibition of the biologic actions of TGF-beta with a neutralizing monoclonal antibody in db/db mice prevents the glomerulosclerosis and renal insufficiency resulting from type 2 diabetes.

Animals↗
PubMed2000

TAJ, a novel member of the tumor necrosis factor receptor family, activates the c-Jun N-terminal kinase pathway and mediates caspase-independent cell death.

We have isolated a novel member of the TNFR family, designated TAJ, that is highly expressed during embryonic development. TAJ possesses a unique cytoplasmic domain with no sequence homology to the previously characterized members of the TNFR family. TAJ interacts with the TRAF family members and activates the JNK pathway when overexpressed in mammalian cells. Although it lacks a death domain, TAJ is capable of inducing apoptosis by a caspase-independent mechanism. Based on its unique expression profile and signaling properties, TAJ may play an essential role in embryonic development.

Amino Acid Sequence↗
PubMed2000

Exposure of preschoolers to lead in the Makati area of Metro Manila, the Philippines.

A study was conducted in the Makati area of Metro Manila, the Philippines. The study evaluated the significance of playground soil lead intake to the total daily lead burden in preschool children. The lead concentration was measured in residential playground soils, food, water, and hair samples by atomic absorption spectrometer and the data were used to draw conclusions. All of the playground soils were contaminated with lead levels ranging from a minimum of 34.54 microg/g to a maximum of 283.13 microg/g in comparison to the naturally occurring lead level of 15 microg/g in soils. Further, the study conducted a lead analysis on the hair of the study population to determine the accumulated lead intake. Ingestion of 25 mg/day of soil contributed to 4% of the total lead intake per day by children exposed to the study sites. Food (83%) was found to be the major source of lead compared to other sources. The study also includes economic valuation and cost benefit analysis from reducing lead in gasoline.

Child, Preschool↗