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Biomedical subjects

S Chakravarti

Publications and source records attributed to S Chakravarti.

At least 37 records · Page 2Linked to original sources
PubMed1995

Perlecan is a component of cartilage matrix and promotes chondrocyte attachment.

Aggrecan, a chondroitin/keratan sulfate-containing proteoglycan, is a major component of cartilaginous tissues. Immunolocalization studies, using antibodies directed to perlecan, a heparan sulfate proteoglycan first detected in basement membranes, and laminin (another major component of basement membranes), indicate that perlecan and laminin are also present in the matrices of hyaline cartilage in the nasal septum, the articular surface of the bone and the growth plate of the developing bone. Consequently, we used antibodies to both aggrecan and perlecan to characterize their synthesis and secretion by primary cultures of chondrocytes derived from the rat chondrosarcoma. Chondrocytes were pulsed for 20 minutes with [35S]methionine and then chased for up to six hours. The radiolabeled perlecan and aggrecan were immunoprecipitated and analyzed by SDS-PAGE. The results show that chondrocytes synthesize precursor proteins to both proteoglycans, but that only the aggrecan precursor protein is secreted as a proteoglycan. Perlecan was also secreted but with less posttranslational modifications than aggrecan. Northern blot analyses of the RNAs from immortalized rat chondrocytes indicated that the major mRNA encoding for perlecan was approximately 13 kb in length, similar in size to that expressed by other cell types, which synthesize 400 kDa core protein perlecan. Analyses of the proteoglycan fractions from the extracts of bovine articular surface indicated that perlecan in this tissue contains both chondroitin and heparan sulfate side-chains. Purified perlecan and laminin were found to promote attachment of immortalized rat chondrocytes in vitro. These studies indicated that perlecan, once thought to be a unique component of the basement membranes, is more widely distributed and is an important component of the cartilage matrix, where it may provide for cell adhesion to the matrix.

Animals↗
PubMed1993

Perlecan gene expression precedes laminin gene expression during differentiation of F9 embryonal carcinoma cells.

F9 embryonal cells can be induced to differentiate and synthesize basement membrane proteins. Perlecan and laminin are two basement membrane constituents that have extensive regions of homology. Expression of perlecan and laminin B1 genes was followed during differentiation of F9 cells by measurements of transcription rate and mRNA abundance using nuclear run on assays and Northern hybridizations, respectively. The rate of precursor protein synthesis was determined by immunoprecipitation from lysates of pulse-labeled F9 cells. The results showed that perlecan gene expression responds more rapidly after induction than does laminin B1 gene expression but is ultimately expressed at a substantially lower level than laminin. Thus, the perlecan and laminin genes appear to be regulated by different mechanisms and their gene products are not made in stoichiometric amounts.

Animals↗
PubMed1990

The basement membrane glycoprotein entactin promotes cell attachment and binds calcium ions.

Mouse entactin derived from the extracellular matrix of M1536-B3 cells and from insect cells infected with a recombinant virus containing entactin sequences were shown to promote the attachment of mouse mammary tumor, human melanoma, and other cells. The cell attachment was inhibited by antibodies against mouse entactin but not by anti-fibronectin or anti-laminin antibodies. On a weight basis entactin was as effective as laminin in promoting the attachment of mouse mammary tumor cells. The attachment of cells to entactin was in part mediated by the integrin recognition RGD peptide sequence. This was demonstrated by the cell attachment properties of peptides derived from entactin which contained this sequence. Furthermore, the peptide RGDS could inhibit the attachment of mouse mammary tumor cells to entactin to approximately 60% of control. It is suggested that additional cell recognition sequences may be present in entactin. The direct binding of calcium ions to entactin was observed. It is probable that the binding sites reside in peptide sequences located toward the NH2 terminus region of entactin. This conclusion was supported by the demonstration that synthetic peptides, containing potential calcium binding sequences derived from entactin, bound calcium. In addition, a recombinant peptide containing the amino-terminal 330 amino acids of entactin also bound calcium ions. The significance of these properties of entactin is discussed.

Amino Acid Sequence↗
PubMed1990

Characterization of the basement membrane glycoprotein entactin synthesized in a baculovirus expression system.

A cDNA clone encoding the entire open reading frame of mouse entactin was constructed. This cDNA was inserted into Autographa californica multiple nuclear polyhedrosis virus and expressed in Spodoptera frugiperda, Sf9, insect cells. The recombinant entactin was produced in large quantities under the control of the polyhedrin promoter. Amino terminus sequence analysis indicated that the signal peptide in the preprotein was correctly cleaved. Polyclonal antibodies prepared against either the naturally occurring or recombinant entactin cross-reacted with both molecules in the native and denatured states. The recombinant molecule promoted cell attachment and exhibited a calcium- and temperature-dependent self-aggregation. These properties reflect those of the naturally occurring mouse molecule. Although entactin is an extracellular matrix component that is normally exocytosed, the vast majority of the molecules were localized inside the cell by immunoelectron microscopy where the molecules formed insoluble aggregates.

Animals↗
PubMed1988

Amino acid sequence and domain structure of entactin. Homology with epidermal growth factor precursor and low density lipoprotein receptor.

Entactin (nidogen), a 150-kD sulfated glycoprotein, is a major component of basement membranes and forms a highly stable noncovalent complex with laminin. The complete amino acid sequence of mouse entactin has been derived from sequencing of cDNA clones. The 5.9-kb cDNA contains a 3,735-bp open reading frame followed by a 3'-untranslated region of 2.2 kb. The open reading frame encodes a 1,245-residue polypeptide with an unglycosylated Mr of 136,500, a 28-residue signal peptide, two Asn-linked glycosylation sites, and two potential Ca2+-binding sites. Analysis of the deduced amino acid sequence predicts that the molecule consists of two globular domains of 70 and 36 kD separated by a cysteine-rich domain of 28 kD. The COOH-terminal globular domain shows homology to the EGF precursor and the low density lipoprotein receptor. Entactin contains six EGF-type cysteine-rich repeat units and one copy of a cysteine-repeat motif found in thyroglobulin. The Arg-Gly-Asp cell recognition sequence is present in one of the EGF-type repeats, and a synthetic peptide from the putative cell-binding site of entactin was found to promote the attachment of mouse mammary tumor cells.

Amino Acid Sequence↗
PubMed1987

Evidence of beta-adrenergic involvement in forced swimming-induced behavioural despair of mice.

Modulation of forced swimming-induced immobility by beta-adrenoceptor activation was investigated in mice. Isoprenaline prolonged the immobility duration of mice in a dose-related manner when the animals were forced to swim for 6 min periods. On the other hand, salbutamol (a beta 2-agonist) reduced the immobility duration of mice. Pretreatment with propranolol (1,2,4,8 and 16 mg/kg, ip), atenolol (10 mg/kg, ip) and metoprolol (10 mg/kg, ip) antagonized the immobility-enhancing effect of isoprenaline. Chronic administration (10 mg/kg/day) for 8 days of propranolol and imipramine protected the animals from isoprenaline-evoked prolongation of immobility. These findings suggest that central beta 1- and beta 2-subtypes of adrenoceptors may be acting in opposite directions to modify the immobility duration of mice. Activation of central beta 1-adrenoceptors may lead to enhanced behavioural despair, whereas a reverse effect may be observed on beta 2-adrenoceptor activation.

Animals↗
PubMed1986

Relationship between cellular RecA protein concentration and untargeted mutagenesis in Escherichia coli.

We measured the production of untargeted mutations in the cI and cII genes of untreated lambda phage undergoing a lytic cycle in UV-irradiated bacterial hosts. As previously shown, treatment with 4 micrograms/ml of rifampicin during post-irradiation incubation inhibited amplification of the RecA protein in these cells. In addition, we observed a decreased mutation rate compared to the untreated, irradiated bacteria. Treatment with 4 micrograms/ml or 8 micrograms/ml rifampicin did not prevent the UV induction of the umuDC operon, as judged by assay of beta-galactosidase activity in a umuC-lacZ fusion strain. In contrast, the UV-induction of beta-galactosidase in the sulA-lacZ fusion strain was decreased by 4 micrograms/ml rifampicin. The inhibition of untargeted mutagenesis by this drug treatment was also observed in a strain constitutive for SOS functions (lexA (Def)) as well as in a RecA-overproducing plasmid strain, suggesting the requirement of other factor(s) in wild-type recA+ cells. An htpR165-carrying strain, that blocks induction of heat-shock proteins, exhibited normal UV-promoted mutagenesis. A correlation was observed between the cellular concentration of RecA protein, increased spontaneously by a temperature shift in a lexA(Ts) strain, and the extent of UV-promoted untargeted mutagenesis. These results suggest a mechanistic role of RecA protein in this process.

Bacterial Proteins↗
PubMed1979

Relation between plasma hormone profiles, symptoms, and response to oestrogen treatment in women approaching the menopause.

Out of a consecutive series of 300 patients seen at a menopause clinic, 82 complained of symptoms generally associated with the climacteric, although they were still menstruating. Vasomotor disturbances were absent in 42 of these patients (group 1) and present in 40 (group 2). Headaches, insomnia, and dyspareunia were the most common complaints among the women with vasomotor symptoms, whereas loss of libido and depression predominated in those without. Conjugated equine oestrogens (Premarin) 1.25 mg daily given for three weeks out of four relieved nearly all symptoms in group 2, but in group 1 the response was disappointing. The mean plasma oestradiol concentration in women with vasomotor symptoms was significantly lower than that observed during days 1-10 of the menstrual cycle, but plasma testosterone values were not significantly different from those observed in younger women. Plasma follicle-stimulating hormone (FSH) and luteinising hormone (LH) concentrations were similar to those seen after the menopause. Concentrations of these hormones in the women without vasomotor symptoms were similar to those in the younger, regularly menstruating women. After six months of oestrogen treatment patients in group 2 had a 2.1-fold increase in mean plasma oestradiol concentration, and plasma FSH and LH concentrations were reduced to 39% and 66% of their pretreatment values respectively; in group 1, however, no such pronounced changes occurred. High concentrations of FSH were present in patients with oestrogen-responsive symptoms, 15 U/1 being the diagnostic cut-off point. This measurement in the presence of characteristic symptoms therefore constitutes the best method of selecting patients for oestrogen-replacement therapy.

Adult↗
PubMed1977

Endocrine changes and symptomatology after oophorectomy in premenopausal women.

The symptomatic and endocrine changes following hysterectomy and bilateral oophorectomy have been studied in 100 patients, 1 to 31 years after surgery. The most frequent symptoms at the time of interview were depression (62 patients), insomnia (48 patients), loss of libido (46 patients) and dyspareunia (38 patients). Vasomotor symptoms were recorded in 28 patients. Although 94 patients claimed that hot flushes were the first symptomatic changes noticed, 60 had complete relief from these symptoms within 6 months of surgery; 34 patients had no symptoms and only 4 patients were taking oestrogen therapy at the time of interview. Plasma oestradiol and testosterone levels were 78 per cent and 27 per cent respectively below the mean values of day 1 to 10 of the menstrual cycle, similar to those found at comparable years after a normal menopause. The plasma FSH level was about 14 times and the plasma LH level about twice the respective preoperative value. Unlike after the normal menopause, these gonadotrophin levels did not show any decline with increasing age. There was no correlation between plasma hormone levels and the presence of vasomotor symptoms or depression.

Castration↗