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Biomedical subjects

S Kochhar

Publications and source records attributed to S Kochhar.

At least 19 recordsLinked to original sources
PubMed2001

Fetal gastrointestinal malformations.

In an audit to evaluate fetal gastrointestinal (GIT) malformations, case sheets of all mothers who gave birth to newborns with GIT malformations were analysed regarding the maternal history, prenatal ultrasound and the postnatal structural malformations and perinatal management. In 1999, eleven babies were born with gastrointestinal malformations, one was a still birth. Nine had associated malformations of other systems, two had trisomy 21. Eight babies were operated in the immediate newborn period, 5 survived. Ultrasound was not very accurate in the diagnosis of GIT anomaly in this series. In view of associated chromosomal and structural anomaly a fetal ECHO and genetic amniocentesis is warranted. Most of these babies require immediate surgery after birth, so they need to be delivered in a centre which is equipped with good neonatal and pediatric surgery care.

Adult↗
PubMed2001

Characterization of peptides formed during fermentation of cocoa bean.

Analysis by SDS-PAGE and GPC-MS of fermented cocoa extracts shows changes in the amount and composition of the major proteins, accompanied by formation of complex distributions of peptides. MS/MS studies and application of SEQUEST sequencing software have allowed identification of two related peptides, a hexapeptide and a nonapeptide, formed from vicilin, one of the cocoa storage proteins. Time course studies of the two peptides show different abundance profiles and indicate, in part, production of the hexapeptide from the nonapeptide.

Cacao↗
PubMed2001

Isolation and characterization of 2S cocoa seed albumin storage polypeptide and the corresponding cDNA.

The amine pool of cocoa is known to be an essential component for the development of the typical cocoa flavor. To better understand and to produce an intense in vitro cocoa flavor, identification of the polypeptides that are the source of the amine flavor precursor pool is essential. Chromatographic analysis of the polypeptide profile of unfermented cocoa resulted in identification of a novel storage polypeptide of M(r) 8515. The N-terminal sequence of the first 34 residues of the purified polypeptide shows similarity to 2S storage albumins of cotton and Brazil nut and sweet protein, Mabinlin. To identify the corresponding cDNA of the putative cocoa 2S albumin, 18 randomly chosen clones from the cDNA library of immature Theobroma cacao seed mRNA were sequenced, and a full-length cDNA clone encoding a protein harboring the N-terminal sequence of the novel polypeptide was selected. The open reading frame of the clone encodes a polypeptide of M(r) 17125. Comparison of the translated amino acid sequence of the precursor protein or the mature polypeptide against the Swiss-Prot and TrEMBL databases shows high sequence similarity (>52%) and identity (>38%) to many plant 2S albumins. Tryptic peptide mass fingerprinting of the purified polypeptide by high-performance liquid chromatography-electrospray ionization mass spectrometry shows 10 masses that match the expected tryptic peptides of the deduced sequence. Together with the published work on plant 2S albumin processing, the results presented here suggest that post-translational processing yields a 73-residue polypeptide (residue positions 78-150) corresponding to the 9 kDa subunit of the mature cocoa 2S albumin protein.

Amino Acids↗
PubMed2001

Sonographic diagnosis of diastematomyelia in utero: a case report and literature review.

Fetal diastematomyelia is a rare form of spinal dysraphism that is characterized by a complete or incomplete division of the spinal cord by an osseous or fibrocartilaginous septum. A case of diastematomyelia, which was detected on the routine third trimester detailed ultrasound scan, is presented. The diagnosis was based on the detection of an echogenic focus in the posterior aspect of the spine in association with widening of the interpedicular vertebral space. The case illustrates that diastematomyelia can occur in the absence of overt spina bifida and that prenatal detection will allow timely postnatal investigation and treatment. Prenatal literature is further reviewed to assess the clinical significance of this finding.

Adult↗
PubMed2000

Primary structure of the abundant seed albumin of Theobroma cacao by mass spectrometry.

The most abundant albumin present in seeds of Theobroma cacao was purified to apparent homogeneity as judged by high-performance liquid chromatography/electrospray ionization mass spectrometry (HPLC/ESI-MS), sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and NH(2)-terminal sequence analysis. Tryptic peptide mass fingerprinting of the purified protein by HPLC/ESI-MS showed the presence of 16 masses that matched the expected tryptic peptides corresponding to 95% of the translated amino acid sequence from the cDNA of the 21 kDa cocoa albumin. Collision-induced dissociation MS/MS analysis of the C-terminal peptide isolated from the CNBr cleavage products provided unequivocal evidence that the mature cocoa albumin protein is nine amino acid residues shorter than expected from the reported cDNA of its corresponding gene. The experimentally determined M(r) value of 20234 was in excellent agreement with the truncated version of the amino acid sequence. The purified cocoa albumin inhibited the catalytic activities of bovine trypsin and chymotrypsin. The inhibition was stoichiometric with 1 mol of trypsin or chymotrypsin being inhibited by 1 mol of inhibitor with apparent dissociation constants (K(i)) of 9.5 x 10(-8) and 2. 3 x 10(-6) M, respectively, for inhibitor binding at pH 8.5 and 37 degrees C. No inhibition of the catalytic activities of subtilisin, papain, pepsin, and cocoa endoproteases was detected under their optimal reaction conditions.

Albumins↗
PubMed2000

Roles of his205, his296, his303 and Asp259 in catalysis by NAD+-specific D-lactate dehydrogenase.

The role of three histidine residues (His205, His296 and His303) and Asp259, important for the catalysis of NAD+-specific D-lactate dehydrogenase, was investigated using site-directed mutagenesis. None of these residues is presumed to be involved in coenzyme binding because Km for NADH remained essentially unchanged for all the mutant enzymes. Replacement of His205 with lysine resulted in a 125-fold reduction in kcat and a slight lowering of the Km value for pyruvate. D259N mutant showed a 56-fold reduction in kcat and a fivefold lowering of Km. The enzymatic activity profile shifted towards acidic pH by approximately 2 units. The H303K mutation produced no significant change in kcat values, although Km for pyruvate increased fourfold. Substitution of His296 with lysine produced no significant change in kcat values or in Km for substrate. The results obtained suggest that His205 and Asp259 play an important role in catalysis, whereas His303 does not. This corroborates structural information available for some members of the D-specific dehydrogenases family. The catalytic His296, proposed from structural studies to be the active site acid/base catalyst, is not invariant. Its function can be accomplished by lysine and this has significant implications for the enzymatic mechanism.

Amino Acid Substitution↗
PubMed1999

Parathyroid cyst: a rare cause of an anterior neck mass.

Parathyroid cysts are rare. Most of them present in the anterior neck as cystic neck swellings. A case of cervical parathyroid cyst is presented, along with a brief review of the literature regarding the aetiology, clinical features, diagnosis and management of this condition.

Adult↗
PubMed1999

Short-chain peptide analysis by high-performance liquid chromatography coupled to electrospray ionization mass spectrometer after derivatization with 9-fluorenylmethyl chloroformate.

Resolution and characterization of short-chain peptides (M(r) = 200-1000) and free amino acids were demonstrated by the use of precolumn derivatization with 9-fluorenylmethyl chloroformate (Fmoc) followed by reverse-phase high-performance liquid chromatography (RP-HPLC) interfaced with an electrospray ionization mass spectrometer (ESI-MS). At pH 10, in addition to derivatization at the N terminus, epsilon-NH(2) and OH groups of lysine and tyrosine residues, respectively, were also derivatized. Fmoc derivatives showed at least 2 orders of magnitude higher ionization potential in the presence of trifluoroacetic acid. The detection levels for both the free amino acid and peptide derivatives were in a few hundred picomoles compared to 10-50 nmol for the underivatized samples. The mass spectra of the peptides before or after derivatization showed the presence of only singly charged ions. However, collision-induced dissociation of the derivatized peptides showed predominance of b-type ions that are relatively less complicated in assigning the peptide sequence.

Amino Acids↗
PubMed1999

Analysis of photocontrol of aspartate kinase in barley (Hordeum vulgare L.) seedlings.

Aspartate kinase (AK) activity is regulated by light. The activity was more in light exposed barley seedlings than those grown in the dark. The light effect was manifested even with small exposures of 5 min duration and red light was more effective than white light in this respect. The effect of 5 min red light could be reversed by a 5 min pulse of far-red light indicating the involvement of phytochrome in this response. The phytochrome is also involved in long term light effects (24 hr exposures with white light). Ca++ takes part in the signal transduction pathway for this light response. Western blot analysis using antibodies raised against the purified lysine- and threonine-sensitive AK isoenzymes from spinach leaves showed no cross reaction with the antibodies to the threonine-sensitive AK in the dark and 5 min far-red light exposed seedlings. But the protein band was detected in the white and red lights. Northern blot analysis of seedlings grown under dark and exposed to white, red and far-red lights and probed with the gene encoding aspartokinase-homoserine dehydrogenase (AKHSD) protein indicated that the gene was differentially expressed. In dark grown seedlings, AKHSD transcript was in low concentration as compared to white light where the transcript concentration was high. A 5 min red light pulse increased the transcript concentration significantly in contrast to 5 min far-red light. The transcript concentration was reduced when 5 min red light was followed by a 5 min far-red light pulse. The AK activity in dark-raised seedlings is attributed to the presence of only one isoenzyme that is sensitive to lysine but insensitive to Ca++ and calmodulin (CAM). In both white and red light exposed seedlings, three isoenzymes of AK were detected. Two of these were sensitive to threonine while one was sensitive to lysine. Both of the threonine sensitive isoenzymes were activated by Ca++ and CAM. Also one of these isoenzymes seems to be located and synthesized in chloroplasts because its synthesis was completely inhibited by chloramphenicol but not by cycloheximide.

Aspartate Kinase↗
PubMed1998

Subunit structure of lysine sensitive aspartate kinase from spinach leaves.

The lysine-sensitive isoenzyme of aspartate kinase was purified to homogeneity from spinach leaves and its subunit composition was studied. The purified preparation had an apparent molecular mass of 280,000 and separated into two subunits- a large subunit with molecular mass of 53,000 and smaller subunit with molecular mass of 17,000 by urea treatment and SDS PAGE. The enzyme molecule has subunit composition of 4 large and 4 small subunits. The activity of the large subunit was stimulated more than two fold by the addition of small subunit and the stimulated activity was inhibited by EGTA. This inhibition could be reversed by Ca++. Further characteristics of the smaller subunit such as heat stability, behavior on ion exchange chromatography, elctrophoretic mobility on polyacrylamide gels, amino acid composition and pattern, presence of trimethyl lysine, its ability to activate other calmodulin stimulated enzymes and its calmodulin-like nature in RIA tests suggested that this subunit is identical to calmodulin.

Aspartate Kinase↗
PubMed1997

Living arrangements of SSI recipients.

This article updates one that appeared in the Bulletin in July 1990. It describes living arrangements of persons receiving payments under the Supplemental Security Income (SSI) program from October 1994 through September 1995. The data were taken from the Quality Assurance review conducted by the Social Security Administration (SSA). This procedure is used by SSA to determine the frequency and causes of incorrect determinations of eligibility and payment amounts. It is difficult to describe the living arrangement for the "typical" recipient. Nevertheless, some interesting patterns emerge in an analysis of the data. About 59 percent (owners and renters combined) of the 6.3 million SSI recipients lived in their own households. Approximately 32 percent of them shared a living arrangement with someone else and about 5 percent of the recipients lived in an institution. Of those SSI recipients living in households, about 36 percent lived alone. Less than 13 percent lived with only their spouses or with only their spouses and minor children. Approximately 11 percent of those in households were child recipients living with parents. An additional 15 percent of the SSI recipients lived in households with only other related adults (other than a spouse or parents).

Adolescent↗
PubMed1996

Lysine-induced premature transcription termination in the lysC operon of Bacillus subtilis.

The expression of the Bacillus subtilis lysC operon, which encodes the first specific enzyme of lysine biosynthesis, is controlled by the availability of the end product, lysine. The question of whether lysine exerts its control by inducing premature termination of transcription was addressed using Northern blot analysis. Whereas lys-C-specific RNA from lysine-starved B. subtilis consisted primarily of the expected full-length mRNA (1.6 kb), that from bacteria grown with an excess of lysine consisted of a truncated 0.27 kb RNA in place of the full-length 1.6 kb transcript. On the other hand, a B. subtilis aecA mutant, in which the lysC operon was derepressed owing to a single nucleotide substitution in the region corresponding to the lysC leader transcript, produced full-length lysC mRNA, but no 0.27 kb RNA, even during growth with excess lysine. Mapping of the truncated 0.27 kb lysC RNA by hybridization with oligonucleotide probes showed that it corresponded to the upstream portion of the lysC leader transcript, extending from the transcription initiation site to a putative rho-independent terminator element. Quantitative transcript analysis by hybridization with specific oligonucleotides showed that lysine did not affect the number of lysC-specific RNA molecules but promoted the stoichiometric replacement of full-length mRNA with truncated 0.27 kb molecules. These results indicate that lysine regulates the expression of the lysC operon by effecting the premature termination of transcription at a rho-independent terminator site in the lysC leader region and that the site of the aecA mutation, far upstream of the putative terminator element, must play an essential role in premature transcription termination by a mechanism which is not yet understood.

Bacillus subtilis↗
PubMed1995

Purification and characterization of homoserine dehydrogenase from spinach leaves.

Homoserine dehydrogenase (HSDH) has been purified to homogeneity from spinach leaves using ammonium sulphate fractionation followed by ion exchange chromatography, gel filtration and FPLC techniques. The purified enzyme has a relative molecular mass of 220,000 and subunit molecular mass of 55,000 and probably occurs as a tetramer. The enzyme was found to be sensitive to threonine and also exhibited aspartate kinase (AK) activity, which was also sensitive to threonine suggesting that it is a bifunctional protein. The enzyme protein also gave a positive cross reaction with antibodies raised against purified AK isoenzymes. Both HSDH and AK activities were stimulated by calcium and calmodulin.

Aspartate Kinase↗
PubMed1995

Disability patterns among SSI recipients.

In December 1993, about 3.8 million persons under age 65 received Supplemental Security Income (SSI) payments because of a disability. More than half of these recipients had some form of mental disorder. In recent years, the number of disabled SSI recipients has climbed sharply. At the same time, there has been a change in the disability patterns among these recipients. The proportion of recipients with mental disorders, particularly those with psychiatric illness, is increasing steadily. Many of these recipients enter the SSI program in their youth and may stay in the program for many years. Similar increases and disability patterns in the Social Security Administration's Disability Insurance (DI) program imply program related causes, including recent changes to the disability requirements and outreach efforts. These changing disability patterns have implications for the size and shape of future SSI caseloads.

Adolescent↗