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Biomedical subjects

K Chandrasekaran

Publications and source records attributed to K Chandrasekaran.

At least 109 records · Page 6Linked to original sources
PubMed1989

Feasibility of identifying amyloid and hypertrophic cardiomyopathy with the use of computerized quantitative texture analysis of clinical echocardiographic data.

Ultrasound tissue characterization, the evaluation of certain physical properties of a tissue based on its acoustic properties, is an evolving application in echocardiography. The ability to identify acutely and chronically injured tissue has been demonstrated in a number of animal studies, but data in humans are limited. The present study tested the hypothesis that quantitative echocardiographic texture analysis, a method of evaluating the spatial pattern of echoes in echocardiographic images, would differentiate amyloid and hypertrophic cardiomyopathy from normal myocardium. Routine clinical echocardiographic data were obtained on 34 subjects at the Mayo Clinic (10 normal subjects, 10 patients with amyloid heart disease, 8 patients with hypertrophic cardiomyopathy and 6 patients with left ventricular hypertrophy due to hypertension). Standard videotape recordings of these echocardiograms were analyzed at the University of Iowa. Echocardiographic data were digitized with use of a calibrated, 256 gray level digitization system. Quantitative texture analysis was performed on data from the ventricular septum and posterior left ventricular wall in end-diastolic and end-systolic, short-axis and long-axis echocardiographic images. The gray level run length texture variables were able to discriminate hypertrophic cardiomyopathy and amyloid heart disease from normal myocardium and from each other (p less than 0.0083 for comparisons of the quantitative texture features of amyloid versus hypertrophic cardiomyopathy versus normal by multivariate analysis of variance). The texture of the myocardium in hypertensive left ventricular hypertrophy not associated with amyloid or hypertrophic cardiomyopathy was in general not significantly different from that of normal myocardium.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1989

Prolongation of ventricular depolarization. ECG manifestation of mexiletine toxicity.

Mexiletine is a type 1B antiarrhythmic drug similar to lidocaine. Prolongation of ventricular depolarization has not been previously reported with the usual oral dosage of mexiletine. We describe a patient with renal failure and heart failure on low-dose oral therapy who developed mexiletine toxicity, which was manifested by ECG prolongation of ventricular depolarization. This was confirmed by elevated plasma concentration of mexiletine. This case illustrates that contrary to the usual belief, mexiletine pharmacokinetics are altered by renal failure. It is important to monitor mexiletine therapy by plasma levels in patients with impaired renal function to avoid mexiletine toxicity.

Aged↗
PubMed1988

Induction of simian virus 40 transplantation immunity in hamsters by gel-purified SV40 large T antigen.

Simian virus 40 (SV40) large T antigen and p53 cellular protein were isolated from an SV40-transformed hamster cell line by immunoprecipitation with anti-T sera and purified by sodium dodecyl sulfate-gel electrophoresis. These two protein were tested in hamsters for the presence of SV40 transplantation rejection antigenic sites by in vivo transplantation rejection assay. The large T antigen immunized the hamsters against a challenge of SV40 tumor cells and the protected animals generated cytotoxic spleen cells. Hamsters immunized with the p53 cellular protein were not protected against SV40-induced tumor but there was some delay in the appearance of tumor.

Animals↗
PubMed1987

Presence of circulating antibodies against cellular protein p53 in a notable proportion of children with B-cell lymphoma.

Serum samples taken from children bearing a wide variety of tumors were screened for the presence of circulating antibodies against the cellular tumor antigen p53. There was a significant correlation (p less than 0.001, n = 119) between the presence of such antibodies and the occurrence of cancer; 12% of the sera tested were positive. These sera were derived from children with a wide range of tumor types. In particular, 21% of the sera obtained from children suffering from a B-cell lymphoma contained anti-p53 antibodies. We were not able to establish a correlation between the secretion of p53-reactive antibodies and any other parameters, such as the age or sex of the child, presence of metastasis, stage or prognosis of disease, or treatment regimen. These results therefore show that the development of p53 immunogenicity is associated with a wide range of neoplastic diseases in children, and in particular with the presence of a B-cell lymphoma.

Adolescent↗
PubMed1986

The transformation-related protein p53 is not bound to the SV40 T antigen in BALB 3T12 cells expressing T antigen.

In most murine cells transformed by the SV40 virus, virtually all of the cellular phosphoprotein p53 is in a complex with the SV40 T antigen. Here, we report that, in SV40-infected T-antigen-positive Balb 3T12 mouse cells, most (approximately 80%) of the p53 is not in complex. Complex formation was determined by measuring the amounts of [35S]methionine-labeled p53 which coprecipitated with T antigen when using monoclonal antibody to T antigen. The amount of complex formation was expressed as a percentage of total p53 present, measured by the amount of p53 precipitated with the monoclonal antibody to the p53. The values were confirmed by Western blotting procedure, in which the steady-state levels of the proteins were measured. In these measurements after complete precipitation with antibody to T antigen, the residual p53 in the supernatant was precipitated by antibody to p53, and this amount was denoted as free p53. There was no significant difference seen between the [35S]methionine-labeled tryptic peptides of complexed and the free p53 (or between complexed and free T antigens) as determined by two-dimensional gel electrophoresis and chromatography. Virus rescue experiments and retransformation by the rescued virus showed that there was no mutation in the SV40 DNA coding for the T antigen which could account for the lack of complex formation. Both p53 and T antigen were underphosphorylated in cells which exhibited reduced complex formation. Tumorigenicity in syngeneic mice and anchorage-independent cell growth in culture of various cloned mouse cells with or without T antigen expression was compared. The changes in the biologic properties were explainable solely on the basis of known or expected effects of expression of the T antigen and were independent of complex formation or of absence of complex formation between p53 and T antigen.

Animals↗
PubMed1986

Echocardiographic visualization of acute myocardial ischemia--in vitro study.

To evaluate whether myocardial texture changes resulting from acute ischemia can be visualized with satisfactory spatial resolution, short axis compound echo images (CEI) (B-scan) were obtained from 12 excised canine hearts in vitro. Seven had myocardial ischemia produced by open chest ligation of the left anterior descending coronary artery (LAD) for 15-30 min prior to excision. The CEI were constructed by compounding 60 simple linear B-scans. Hearts were sectioned after scanning, and gross morphological changes were recorded. Microscopic comparison between grossly abnormal and normal regions were recorded. The CEI from the ischemic group revealed altered myocardial texture seen as bright coarsely granular echoes in the regions normally perfused by the ligated LAD artery. Corresponding anatomic sections revealed increased redness in these regions. Microscopically these regions revealed interstitial and intercellular edema as compared to the normal regions. Acute myocardial ischemia can be visualized in CEI and these regions have significantly increased backscatter, decreased attenuation, and decreased speed of ultrasound relative to normal regions in the same hearts. Myocardial edema is probably responsible for these changes.

Animals↗
PubMed1985

Epithelial HBL-100 cell line derived from milk of an apparently healthy woman harbours SV40 genetic information.

The epithelial HBL-100 cell line was established in vitro from milk of an apparently healthy woman. It exhibits characteristics of transformation from the very beginning and evolves during in vitro maintenance, until becoming tumorigenic in nude mice. This immortal cell line represents a useful model for studying the progression of human epithelial cells toward malignancy. In the course of our investigations we detected a 94K protein in HBL-100 cells obtained from four different sources. This protein is shown to be indistinguishable from the SV40 large T-antigen on the basis of: Recognition by polyclonal and different monoclonal antibodies. Partial peptide map analysis. Specific binding capacity to the SV40 DNA origin of replication. The presence of a tandemly integrated SV40 genome is demonstrated by Southern blotting. Successful rescue of SV40 DNA by fusion with permissive COS-7, but not CV-1 cells, indicates that the SV40 T-antigen from HBL-100 cells is defective in a function(s) essential to the replication of the viral DNA. The possible origin of the SV40 genetic information that we have detected in HBL-100 cells and the implications of this finding on studies involving this cell line are discussed.

Adenocarcinoma↗
PubMed1985

P53-transformation-related protein: kinetics of synthesis and accumulation in SV40-infected primary mouse kidney cell cultures.

During abortive infection of Go/G1-arrested primary baby mouse kidney (BMK) cell cultures with simian virus 40 (SV40), expression of the viral large T antigen is followed by a mitotic host response including the stimulation of host macromolecular synthesis and induction into the cell cycle of Go/G1-arrested cells. We performed an extensive study of the sequential events taking place after SV40 infection of confluent BMK cell cultures. This study comprised a detailed kinetic analysis of transcription, synthesis, and accumulation of p53, in conjunction with the time course of large T antigen synthesis and SV40-induced cellular DNA replication. The monoclonal antibodies used for specifically recognizing mouse p53 were PAb 421, PAb 122, PAb 246, PAb 248, and RA3-2C2. Our results consistently show that under our experimental conditions, the stimulation of p53 synthesis and the accumulation of p53 occur well after the onset of T antigen-induced cellular DNA replication. This relatively late activation of p53 expression appears to be controlled at a level other than transcription. In conclusion, we suggest that, at least in certain cases, T antigen's mitogenic potential is not dependent on its interaction with p53.

Animals↗
PubMed1983

Immunoaffinity chromatography of a cellular tumor antigen from mouse neuroblastoma cells.

The cellular tumor antigen p53 was isolated from mouse neuroblastoma cells and was found in a form not complexed to another protein. The p53 in these cells was stable, turning over about every 10 h. Its methionine-labelled tryptic peptides were very similar to those of the p53 isolated from SV40-transformed mouse cells. The labelled protein was purified from neuroblastoma cells by immunoaffinity using specific monoclonal antibodies and was about 80% radiochemically pure. Furthermore, the purified p53 sedimented in sucrose gradients with a sedimentation coefficient of approximately 8S. This correlated with the sedimentation coefficient of p53 prior to purification, showing that the purified protein retained its native size.

Animals↗
PubMed1982

The amount of a specific cellular protein (p53) is a correlate of differentiation in embryonal carcinoma cells.

A specific cellular protein of molecular weight of 53-55,000 (p53) has been shown to be induced in all SV40 transformed cells. A similar protein has also been shown to be present in embryonal carcinoma cells and in midgestation murine embryo primary cells, which are not infected by SV40. In embryo cell primaries the amount of the protein was shown to decrease with the increase in the stage of embryo development. As differentiation or decrease in cell growth rate can account for this, and since the growth rate of embryo primary cells cannot be measured, we chose to investigate various embryonal carcinoma cells. We report that the p53 is present in a pluripotent embryonal carcinoma cell OTT6050, and in its differentiated parietal endoderm derivative, PYS-2 cells. The amount of p53 is higher in the undifferentiated EC stem cells than in the differentiated PYS-2 (parietal endoderm) cells. The amount of the protein decreases in F9 embryonal carcinoma cells induced to differentiate to a parietal endoderm cell type by treatment with retinoic acid, as it does following spontaneous differentiation of OTT6050 EC cells. To determine if a change in growth rate, rather than differentiation, might account for the diminished levels of this protein, the amount of p53 was measured in growing and in growth arrested cell populations. When the growth rate of F9 cells was reduced by treatment with 8-bromocyclic AMP there was no change in the amount of p53. The half life of the p53 was compared in the undifferentiated and the differentiated cell types to determine if a change in stability might account, in part, for the altered levels of this protein. The p53 is found to be most stable in the SV40 transformed established clonal cells. It is less stable in the fibroblast clonal cells which were not transformed by SV40. The results of these experiments indicate that a decrease in the amount of p53 primarily correlates with differentiation in the embryonal carcinoma cell lines studied and not with cell growth rate. Furthermore, the decrease appears to be related (in part) to the decreased stability of the p53.

Animals↗
PubMed1982

Quantitation of a 55K cellular protein: similar amount and instability in normal and malignant mouse cells.

Quantitative expression of a specific 55,000 (55K)-molecular-weight cellular protein was studied in two groups of mouse embryo fibroblast (clonal) cells originating from two parent clones, one of which possessed high tumorigenicity and the other of which possessed very low tumorigenicity. From the clone with low tumorigenicity, tumor lines and clones were obtained by selecting rare spontaneously transformed highly tumorigenic (mutant) cells. Cells were labeled during exponential growth for 3 h at 37 degrees C, with [35S]methionine, and the cellular 55K protein was immunoprecipitated with a monoclonal antibody and quantitated. There were low and approximately equal amounts of 55K protein in cells (clones) with both low and high tumorigenicity from both groups of cells, and there was no correlation at all between quantitative expression of 55K protein and of cellular tumorigenicity. There was approximately 10- to 20-fold more 55K protein in all simian virus 40-transformed T antigen-positive derivative clones, as shown previously. The T antigen-negative revertant tumor lines and clones obtained by an immunological in vivo selection method had low amounts of 55K protein, similar to the parent cell before simian virus 40 transformation. In all of the T antigen-negative cells, including the highly tumorigenic cells, degradation (turnover?) of the 55K protein was rapid, and a half-life of 15 to 60 min was estimated from pulse-chase experiments. In all of the T antigen-positive cells the 55K protein was stable (half-life greater than 10 h). In primary cells established from the tumors induced by highly tumorigenic cells there was a very low or no detectable amount of the 55K protein. This is in contrast to the primary cells obtained from early murine embryos in which we have reported high amounts of (stable) 55K proteins.

Animals↗
PubMed1981

Surface proteins of simian-virus-40-transformed cells.

Mammalian cells transformed in tissue culture by SV40 were shown to contain, in addition to the SV40-coded 94,000 d large T antigen and the 20,000 d small t antigen, a approximately 56,000 d cellular protein, which specifically precipitates with sera of animals bearing SV40-induced tumor(s) (tumor or T serum). We investigated the presence of these three proteins at the surface of logarithmically growing SV40-transformed cloned mouse cells, after metabolic labelling with [35S]-methionine for 3 h. The 56,000 d protein was found to be susceptible to digestion by trypsin under conditions which did not disrupt the cells, while no small t antigen was found to be digested. Both the 56,000 d cellular protein and the SV40 large T antigen were susceptible to lactoperoxidase-catalyzed iodination from the outside of intact cells. Trypsin treatment removed both the iodinated 56,000 d protein and the iodinated SV40 large T antigen. These experiments indicated that (a certain amount of) the 56,000 d protein and a relatively small amount of the large T antigen (which is present mainly in the nucleus) are present on the cell surface. The results confirm and extend independent experiments using subcellular fractionation techniques (Luborsky and Chandrasekaran, 1980; Soule and Butel, 1979). After heat treatment (at 50 degrees C for 30 min) of the whole-cell extract the 56,000 d cellular protein was precipitated by the tumor serum in the absence of precipitation of SV40 large T antigen. This result showed that the 56,000 d protein is more (thermo)stable (in the whole-cell extract) than the SV40 large T antigen, and also indicated that the tumor serum employed had antibodies against the 56,000 d cellular antigen. The heat-treated whole-cell extract of Sv40-transformed mouse cells was able to immunize and fully protect mice against a lethal tumorigenic dose of SV40-transformed cells. These results suggest the need for further experiments to characterize the chemical and immunologic properties of the 56,000 d protein.

Animals↗
PubMed1981

Quantitation and characterization of a species-specific and embryo stage-dependent 55-kilodalton phosphoprotein also present in cells transformed by simian virus 40.

A 55-kilodalton (kDal) protein was detected recently in primary cultures of day 12 mouse embryos by immunoprecipitation with serum from simian virus 40 (SV40) tumor-bearing hamsters (T serum), Preliminary evidence suggested that this protein was similar to a cellular 55-kDal protein induced after SV40 transformation of mouse cells. We now show that specific approximately 55-kDal [35S]methionine-labeled proteins precipitate from primary cultures of midgestation mouse, rat, and hamster embryos on addition of T serum or monoclonal antiserum prepared against the SV40-induced mouse 55-kDal proteins. The two-dimensional maps of the [35S]methionine-labeled tryptic peptides of the mouse, hamster, and rat embryo proteins are similar to the maps of the corresponding proteins from SV40-transformed cells. Primary cells from midgestation mouse, hamster, or rat embryos contain one-third to one-half as much 55-kDal protein as a SV40-transformed mouse fibroblast cell and nearly the same amount as F9 mouse embryonal carcinoma cells. The amount of 55-kDal protein is greatly reduced on replating the mouse, rat, or hamster embryo primary cells. The amount of this protein in mouse embryos is dependent on the stage of the embryo. The embryo proteins are phosphoproteins.

Animals↗
PubMed1980

An embryo protein induced by SV40 virus transformation of mouse cells.

A specific protein of molecular weight (MW) approximately 55,000 (55K) was found recently by immunoprecipitation in all SV40 virus-transformed mammalian cells, in addition to the SV40 large T antigen (appoximately 94K) and small antigen (approximately 17K), which are the only proteins coded by the 'early half' of the SV40 genome. The 55K protein is encoded by cellular DNA; its peptide pattern is different from that of the SV40 antigens and it is species specific in mouse, rat, hamster, monkey and human SV40-transformed (or infected) cells. A 55K protein with a similar peptide pattern was found in mouse embryonal carcinoma cells not exposed to SV40. Similar proteins were reported in mouse sarcomas and leukaemias induced by a great variety of aetiological agents and also in a spontaneously transformed mouse fibroblast cell line, and it has been suggested that the protein may be a general correlated of cellular tumorigenicity. We now report that the approximately 55K protein is present in primary cell cultures from 12-14 day old mouse embryos, but not in 16-day old mouse embryos. The embryo protein has a peptide pattern virtually indistinguishable from that of the SV40-induced protein. We also show by comparing closely related cell families that spontaneously transformed highly tumorigenic mouse cells do not possess the 55K protein.

Animals↗