PubMed Health⌕ Search

Biomedical subjects

J Sharma

Publications and source records attributed to J Sharma.

At least 37 records · Page 2Linked to original sources
PubMed2000

I-cell disease (Mucolipidosis II).

I-cell disease (Mucolipidosis II) is one of the lysosomal storage diseases which presents in the neonatal period, and within six months will phenotypically resemble the severe forms of the group of disorders called the "mucopolysaccharidoses" but without mucopolysacchariduria. In Mucolipidosis II, fibrocytes exhibit "abnormal lysosomes". Activities of several lysosomal enzymes are low in fibroblast cultures but high in mucolipidosis II serum. We present a patient with I-cell disease diagnosed on the basis of clinical, radiological and biochemical features. The mother of this child was pregnant and the fetus was also found to be affected.

Child, Preschool↗
PubMed2000

Adaptation-induced plasticity of orientation tuning in adult visual cortex.

A key emergent property of the primary visual cortex (V1) is the orientation selectivity of its neurons. The extent to which adult visual cortical neurons can exhibit changes in orientation selectivity is unknown. Here we use single-unit recording and intrinsic signal imaging in V1 of adult cats to demonstrate systematic repulsive shifts in orientation preference following short-term exposure (adaptation) to one stimulus orientation. In contrast to the common view of adaptation as a passive process by which responses around the adapting orientation are reduced, we show that changes in orientation tuning also occur due to response increases at orientations away from the adapting stimulus. Adaptation-induced orientation plasticity is thus an active time-dependent process that involves network interactions and includes both response depression and enhancement.

Adaptation, Physiological↗
PubMed1999

Rewiring cortex: the role of patterned activity in development and plasticity of neocortical circuits.

Visually driven activity is not required for the establishment of ocular dominance columns, orientation columns, and long-range horizontal connections in visual cortex, although spontaneous activity appears to be necessary. The role of activity may be instructive or simply permissive; evidence for an instructive role requires inquiry into the role of the pattern of activity in shaping cortical circuits. The few experiments that have probed the role of patterned activity include the effects of artificial strabismus, artificial stimulation of the optic nerve, and rewiring visual projections from the retina to the auditory thalamus and cortex. These experiments demonstrate that patterned activity is vital for the maintenance of thalamocortical, local intracortical, and long-range horizontal connections in cortex.

Animals↗
PubMed1998

Isolation and characterization of monomeric and dimeric CP47-reaction center photosystem II complexes.

Using the detergents n-dodecyl beta-D-maltoside and heptyl thioglycopyranoside, a subcore complex of photosystem II (PSII) has been isolated that contains the chlorophyll-binding protein, CP47, and the reaction center components, D1, D2, and cytochrome b559. We have found, by using sucrose density centrifugation, that the resulting preparation consisted of a mixture of dimeric and monomeric forms of the CP47 reaction center (RC) complex, having molecular masses of 410 +/- 30 and 200 +/- 28 kDa, respectively, as estimated by size exclusion chromatography. The level of the dimer in the preparation is significantly higher than the monomeric form. Both the monomer and dimer contain the proteins CP47, D1, and D2 and the alpha- and beta-subunits of cytochrome b559. Analyses by mass spectrometry and N-terminal sequencing showed that both forms of the CP47-RC complex contain the products of the psbI, psbTc (chloroplast gene), and psbW with molecular masses of 4195.5, 3849.6, and 5927.4 Da, respectively. In contrast to the monomeric form, the CP47-RC dimer contained two extra proteins with low molecular weights, identified as the products of the psbL and psbK genes having molecular masses of 4365.5 and 4292.1, respectively. It was also found that the dimer contained slightly more molecules of chlorophyll a (21 +/- 2.5) than the monomer (18 +/- 1.5), a characteristic also observed in the room temperature absorption spectrum by comparing the ratio of absorption at 416 and 435 nm. Of particular note was the finding that the dimer, but not the monomer, contained plastoquinone-9 (estimated to be 1.5 +/- 0.3 molecules per RC). The results indicate that the CP47-RC monomer is derived from the dimeric form of the complex, and therefore the latter is likely to represent an in vivo conformation. The PsbTc as well as the PsbI and PsbW proteins are identified as being intimately associated with the D1 and D2 proteins, and in the case of the dimer, importance is placed on the PsbL and PsbK proteins in sustaining plastoquinone binding and maintenance of the dimeric organization. Assuming only one copy of the alpha- and beta-subunits of cytochrome b559, the monomeric and dimeric forms of the complex would be expected to contain 21 and 23 x 2 transmembrane helices, respectively.

Chromatography, High Pressure Liquid↗
PubMed1997

Purification and determination of intact molecular mass by electrospray ionization mass spectrometry of the photosystem II reaction center subunits.

A reverse phase high pressure liquid chromatography purification system for the rapid separation of photosystem II reaction center proteins free of salts and detergents is described. This procedure results in the isolation of the three small subunits: alpha- and beta-subunits of cytochrome b559 and PsbI protein, with near base-line resolution between each peak, although the D1 and D2 proteins were partially deconvoluted. The molecular masses obtained by electrospray ionization mass spectrometry for the purified beta-subunit of cytochrome b559, alpha-subunit of cytochrome b559, and the PsbI protein, 4,394.8 +/- 0.4, 9,283.7 +/- 0.8, and 4,209.5 +/- 0.4 Da, respectively, are in excellent agreement with values obtained from previous characterization studies (Sharma, J., Panico, M., Barber, J., and Morris, H. R. (1997) J. Biol. Chem. 272, 3935-3943). Direct electrospray analysis of the D1 and D2 proteins suggests that these components exist in heterogeneous forms. The molecular mass ascribed to a predominant form of the D1 protein, 38, 040.9 +/- 6.5 Da, and the D2 protein, 39,456.1 +/- 7.7, are also in agreement with those expected for the mature nonphosphorylated states of these subunits.

Chromatography, High Pressure Liquid↗
PubMed1997

Primary structure characterization of the photosystem II D1 and D2 subunits.

Mass spectrometry techniques have been applied in a protein mapping strategy to elucidate the majority of the primary structures of the D1 and D2 proteins present in the photosystem II reaction center. Evidence verifying the post-translational processing of the initiating methionine residue and acetylation of the free amino group, similar to those reported for other higher plant species, are presented for the two subunits from pea plants (Pisum sativum L.). Further covalent modifications observed on the D1 protein include the COOH-terminal processing with a loss of nine amino acids and phosphorylation of Thr2. In addition, the studies reported in this paper provide the first definitive characterization of oxidations on specific amino acids of the D1 and D2 proteins. We believe that these oxidations, and to a much lesser extent the phosphorylations, are major contributors to the heterogeneity observed during the electrospray analysis of the intact subunits reported in the accompanying paper (Sharma, J., Panico, M., Barber, J., and Morris, H. R. (1997) J. Biol. Chem. 272, 33153-33157). Significantly, all of the regions that have been identified as those particularly susceptible to oxidation are anticipated (from current models) to be in close proximity to the redox active components of the photosystem II complex.

Amino Acid Sequence↗
PubMed1997

Aortic stenosis in hypoplastic right heart syndrome, associated with interstitial deletion of chromosome 2.

This report describes a rare case of moderate to severe valvular aortic stenosis in the setting of severe tricuspid stenosis, pulmonary atresia with intact interventricular septum and right ventricular dependent coronary circulation. We discuss the clinical, hemodynamic, echocardiographic and angiocardiographic findings on this rare case and comment on accompanied malformations, with its possible association observed to the interstitial deletion of band q22 or q24 of the long arm of chromosome 2.

Abnormalities, Multiple↗
PubMed1997

Characterization of the low molecular weight photosystem II reaction center subunits and their light-induced modifications by mass spectrometry.

A sensitive and simple reverse phase HPLC purification scheme was developed for the rapid separation of the small protein subunits from photosystem II reaction center preparations. The precise molecular masses of the alpha- and beta-subunits of cytochrome b559 and the psbI gene product from pea plants, found to be 4394.6 +/- 0. 6, 9283.6 +/- 0.7, and 4209.5 +/- 0.5 Da, respectively, were then successfully determined for the first time by electrospray- and fast atom bombardment-mass spectrometry. Discrepancies between the molecular weights assigned and those calculated from the respective DNA sequences were observed for alpha- and beta-subunits of cytochrome b559. Currently, the nucleotide sequence of the psbI gene product from pea plants is not available. Application of novel mapping and sequencing strategies has assured the elucidation of full primary structures of all of the purified subunits. The modifications identified here include the post-translational processing of the initiating methionine on both subunits of cytochrome b559, NH2-terminal acetylation and an mRNA editing site at residue 26 (Ser --> Phe) on the beta-subunit, and retention of the NH2-terminal formyl-Met on the psbI gene product. In addition, specific oxidation of a single amino acid residue was identified on the psbI gene product and the beta-subunit purified from light-treated reaction center preparations. Overall, these studies provide the first detailed primary structural characterization of the small subunits of the reaction center complex and their associated light-induced modifications.

Amino Acid Sequence↗
PubMed1996

Orientation maps of subjective contours in visual cortex.

Responses to subjective contours in visual cortical areas V1 and V2 in adult cats were investigated by optical imaging of intrinsic signals and single-unit recording. Both V1 and V2 contain maps of the orientation of subjective gratings that have their basis in specific kinds of neuronal responses to subjective orientations. A greater proportion of neurons in V2 than in V1 show a robust response to subjective edges. Through the use of subjective stimuli in which the orientation of the luminance component is invariant, an unmasked V1 response to subjective edges alone can be demonstrated. The data indicate that the processing of subjective contours begins as early as V1 and continues progressively in higher cortical areas.

Animals↗
PubMed1995

Sequence and unusual 3' flanking region of the rat tRNA[Ser]Sec gene.

The single rat selenocysteine tRNA (tRNA[Ser]Sec) locus, including flanking sequence, was isolated by molecular cloning and its nucleotide (nt) sequence determined. In addition to the identification of likely regulatory elements 5' of this gene, this analysis also revealed a novel 3' repeat element consisting of three and a half repetitions of a 34-nt unit.

Animals↗
PubMed1995

Correlative histochemical and biochemical studies on acetylcholinesterase activity during ovulation in the rat.

A correlative histochemical and biochemical study has been made of the changes in acetylcholinesterase (AChE)-positive sites during ovulation induced in immature female albino rats with exogenous gonadotrophins. Acetylcholinesterase is the marker for cholinergic innervation. In histochemical observations, AChE+ve sites were localized in typical perivascular position and the intensity and number of enzyme-positive sites around the Graafian follicle gradually increased during follicular maturation and ovulation but stromal AChE-activity did not show any appreciable change. Biochemical estimation of the enzyme supports these observations. The electronmicroscopic and histological observations of the structural changes in nerves accompanying ovulation have also been enumerated and the role of cholinergic nerves in neuromuscular control of ovulation is discussed.

Acetylcholine↗
PubMed1994

Prognostic significance of insulin-like growth factor-binding protein expression in axillary lymph node-negative breast cancer.

BACKGROUND: Cellular proliferation, as measured by S-phase fraction, is an important predictor of breast cancer prognosis. The insulin-like growth factors (IGFs) have been shown to regulate proliferation in both normal and neoplastic cells by interacting with specific cell surface receptors. In addition to these receptors, high-affinity extracellular binding proteins also modulate IGF action. These insulin-like growth factor-binding proteins (IGFBPs) could influence breast cancer growth and, like other biological parameters of proliferation, could be related to prognosis. PURPOSE: To test whether IGFBP expression was related to other biological parameters and disease-free survival, we measured IGFBP expression in 238 lymph node-negative primary breast cancer specimens. METHODS: Proteins were extracted from breast cancer specimens and analyzed by semiquantitative IGF-I ligand blotting for IGFBP expression. IGFBP expression levels were compared to tumor size, age, S-phase fraction, DNA ploidy, and estrogen and progesterone receptor expression by Spearman correlation. RESULTS: Binding protein (BP)-2, BP-3, BP-4, and BP-5 were identified in breast cancer extracts. Estrogen receptor expression was positively correlated with BP-2 (Spearman correlation coefficient, rs = .262; P = .0001), BP-4 (rs = .313; P = .0001), and BP-5 (rs = .242; P = .0002). Similar correlations between progesterone receptor and BP-2, BP-4, and BP-5 were also found. BP-3 was inversely correlated with age (rs = -.251, P = .0001). BP-4 was weakly inversely correlated with tumor size (rs = -.141; P = .0295) and S-phase fraction (rs = -.216; P = .0025). Since tumor size and S-phase fraction are powerful predictors of prognosis in node-negative breast cancer, we examined the value of BP-4 as a predictor of disease-free survival. When stratified by tumor size, patients with large (> 2 cm) tumors that expressed low levels of BP-4 had improved survival when compared with patients with large tumors and high BP-4 levels (P = .001). CONCLUSIONS: IGFBPs can be detected in breast cancer specimens, and their level of expression correlates with other known biological parameters of breast cancer. Large tumors with low levels of BP-4 have relatively favorable prognoses. IMPLICATIONS: These data suggest that the IGFBPs may play a role in breast cancer biology and that BP-4 levels, analyzed in conjunction with tumor size, may have prognostic significance.

Breast Neoplasms↗
PubMed1994

Use of N,N'-polymethylenebis(iodoacetamide) derivatives as probes for the detection of conformational differences in tubulin isotypes.

Mammalian brain tubulin is an alpha beta heterodimer; both alpha and beta exist in 6-7 isotypic forms which differ in their amino acid sequences. By the use of isotype-specific monoclonal antibodies, we have previously shown that we can purify the alpha beta II, alpha beta III, and alpha beta IV tubulin dimers from bovine brain. We have also observed that these isotypes differ in their distribution in vivo and their polymerization and drug-binding properties in vitro. We have now explored the question of whether the isotypically purified dimers differ in their overall conformation using as probes compounds of the N,N'-polymethylenebis (iodoacetamide) series which are known to form discrete intrachain cross-links in beta-tubulin. These compounds have the structure ICH2CONH(CH2)nNHCOCH2I. One of these cross-links, designated beta s, is between cys12 and either cys201 or cys211. The other, designated beta*, is between cys239 and cys354. The beta* cross-link forms in alpha beta II and alpha beta IV but not in alpha beta III; this is not surprising in view of the fact that alpha beta III has serine at position 239 instead of cysteine. However, alpha beta III is also unable to form the beta s cross-link, although it appears to have all three cysteines which may be involved in the cross-link. This suggests that at least one of the sulfhydryls involved in the cross-link may be inaccessible in alpha beta III. Although both alpha beta II and alpha beta IV can form the beta s cross-link, the dependence on cross-linker chain length is different. alpha beta II forms beta s with derivatives in which n = 2, 4, 5, 6, and 7 but not with those in which n = 3 or 10. In contrast, alpha beta IV forms beta s with derivatives in which n = 2, 3, 4, 5, 6, 7, and 10. These results imply that the beta s sulfhydryls are slightly more accessible in alpha beta IV and are therefore less dependent on the conformation of the cross-linker to react with it. It appears, therefore, that the alpha beta II, alpha beta III, and alpha beta IV dimers each have unique conformations. This may help to explain the different assembly and drug-binding properties of these dimers.

Animals↗
PubMed1994

Improved renal preservation with PB-3 flush solution during 72 h of cold storage.

Previously, PB-2 flush solution has been found to be superior to Collins 2 solution (C-2) in extending renal viability in the dog. To further characterize preservation mechanisms, we studied mitochondrial oxidative function during 72 h of cold storage comparing PB-3, UW-1, and C-2 flush storage solutions. Complex 1 dependent mitochondrial oxidative phosphorylation (MOP) was found to be significantly less (P < 0.001) at 5 h and 72 h of cold storage (P < 0.001) for the C-2 compared to the other flush groups. Complex 2 dependent MOP had parallel results, having significantly less function (P < 0.002) at 5 h and 72 h (P < 0.02) of cold storage in the C-2 group compared to the other groups. PB-3 and UW-1 solutions were noted to be comparable, suggesting possible equivalent preservation efficacy. Nevertheless the components of PB-3 at the level of MOP contributed significantly to better preservation compared to C-2 solution.

Adenine Nucleotides↗