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Biomedical subjects

A Sen

Publications and source records attributed to A Sen.

At least 145 records · Page 8Linked to original sources
PubMed1992

Formation of a new stable phase of phosphatidylglycerols.

Dilauroyl and dimyristoylphosphatidylglycerol (DMPG) form a more stable gel state when aqueous suspensions are incubated several days at low temperature (0-2 degrees C), pH 7.4 with 0.15 M NaCl. This gel state is characterized by a higher transition temperature and a higher transition enthalpy. The geometry of this gel state is distinguishable from the metastable gel state that forms rapidly upon hydration on the basis of its x-ray diffraction pattern. Infrared spectra in the CH2 scissoring region indicate that the stable gel phase of DMPG is also characterized by reduced reorientational fluctuations of acyl chains and increased interchain interactions. Analysis of vibrational bands due to ester carbonyl groups of DMPG suggests that the transition to a new gel phase is initiated by changes in the interfacial and/or headgroup region of the bilayer, most likely via formation of interlipid hydrogen bonds. The melting of the stable gel phase of DMPG is accompanied by a gross morphological change resulting in vesiculation.

Calorimetry, Differential Scanning↗
PubMed1992

The effects of hypocalcemia/hypophosphatemia on porcine bone and dental hard tissues in an inherited form of type 1 pseudo-vitamin D deficiency rickets.

The effects of Vitamin D deficiency rickets on the formation of mineralized dental tissues were studied in a breed of pigs which had moderate and marked hypocalcemia because of an inherited defect in the renal production of the biologically-active Vitamin D metabolites. Affected piglets developed classical symptoms of rickets which were fatal unless Vitamin D supplements were given. The dissected mandibles of homozygous (rickets) and heterozygous (normal) pigs were photographed and radiographed. Compared with those of normal pigs, the mandibles of homozygous pigs demonstrated slowed development/eruption of permanent teeth, under-mineralized bone, underdeveloped dentin (enlarged pulp chambers), interglobular dentin, and enamel hypoplasia. Enamel defects in rachitic pig teeth were difficult to observe radiographically, but could be detected visually and by SDS-PAGE analysis of the enamel protein components of developing and maturing enamel. There was significant retention of amelogenins in the enamel maturation zones of developing molars taken from rachitic pigs, but there was no obvious difference in the patterns of amelogenin processing.

Amelogenin↗
PubMed1991

Environmental signals induce major changes in virulence of Shigella spp.

Growth conditions play a major role in expression of virulence by Shigella spp. both in vitro (adherence and internalization in eukaryotic host cells) and in vivo (keratoconjunctivitis). Optimized expression of virulence required anaerobic growth to log phase in particular media such as brain heart infusion broth. Kinetic studies of guinea pig eye infections showed that as few as 2 x 10(5) S. dysenteriae CG097 or S. flexneri M90T, grown under these optimized conditions, produced keratoconjunctivitis in 15 h. In vitro studies demonstrated that adherence to and invasion of Henle 407 cells, at 37 degrees C, by organisms grown under these optimized conditions, were significantly greater than when organisms were grown aerobically under the same conditions.

Anaerobiosis↗
PubMed1991

Molecular organization and stability of hydrated dispersions of headgroup-modified phosphatidylethanolamine analogues.

Measurements of the thermotropic behavior of various headgroup-modified analogues of 1-palmitoyl-2-oleoylphosphatidylethanolamine (POPE) and of the ion-triggered destabilization of unilamellar vesicles containing these species have been correlated with X-ray diffraction measurements of the organization of hydrated dispersions of these analogues in the absence and presence of dodecane. The hexagonal II lattice repeat dimension -dhex for dodecane-supplemented dispersions, which reflects the optimal or "spontaneous" radius of surface curvature of the phospholipid component, is increased relative to POPE for most analogues with N-alkyl substitutents or increased amino-to-phosphate group separations. Interestingly, however, POPE analogues that are alkylated on C-1 or C-2 of the ethaolamine group show smaller -dhex values (and hence smaller spontaneous radii of surface curvature) than does POPE itself, despite the greater steric bulk of their headgroups. The lamellar-to-hexagonal II transition temperatures of the various POPE analogues and their abilities to promote contact-dependent vesicle destabilization both show strong correlations with the analogues' measured -dhex values (and hence with their spontaneous radii of curvature). The uniformity of these correlations over a wide range of headgroup structures strongly supports, and may help to refine, recent theories which postulate that the spontaneous surface curvature of a lipid or lipid mixture is a central, quantitative determinant of its tendency to adopt nonlamellar phases and to undergo contact-dependent bilayer destabilization.

Calorimetry↗
PubMed1991

Bilayer packing stress and defects in mixed dilinoleoylphosphatidylethanolamine and palmitoyloleoylphosphatidylcholine and their susceptibility to phospholipase A2.

The hydrolysis of mixed dilinoleoylphosphatidylethanolamine (DiLinPE) and 1-palmitoyl-2-oleoylphosphatidylcholine (POPC) dispersions by porcine phospholipase A2, under conditions leading to the bilayer-to-nonbilayer phase transition, has been studied. Two structurally distinct forms of the dispersions were used, multilamellar vesicles (MLV) and supercritical large unilamellar vesicles (SCLUV). In MLV, maximum free fatty acid was produced in dispersions containing 85 mol % DiLinPE. The peak in the fatty acid release is found at the onset of appearance of the nonbilayer defects reported earlier. DiLinPE was found to be preferentially hydrolyzed as compared to POPC. When cholesterol was added to the mixed DiLinPE/POPC MLV, the onset of the observable appearance of nonbilayer defects, the positions of the peaks for total hydrolysis, and the preferential hydrolysis of DiLinPE were all shifted toward lower DiLinPE concentrations. In SCLUV, where the appearance of nonbilayer structures is prevented by constraining the lipids in bilayer configuration, the hydrolysis by PLA2 increases with increasing DiLinPE as predicted from the increase in the calculated monolayer bending energy. The results are interpreted to be related to the pretransition molecular-packing stress and defects at the onset of the bilayer-to-nonbilayer transition. Results indicate that the porcine pancreatic phospholipase A2 activity is controlled by bilayer-packing stress, which may cause structural defects of the substrate, among other factors. Results also indicate a preferential localization of PE at stress-related defect regions.

Animals↗
PubMed1991

New insights into the mitochondrial carnitine palmitoyltransferase enzyme system.

Dissection of the mitochondrial carnitine palmitoyltransferase (CPT) enzyme system in terms of its structure/function relationships has proved to be a formidable task. Although no one formulation has gained universal agreement we believe that the weight of evidence supports a model with the following features: a) in any given tissue CPT I and CPT II are distinct proteins; b) CPT I, unlike CPT II, is detergent labile; c) within a species CPT II is expressed body wide, whereas CPT I exists as tissue specific isoforms; d) malonyl-CoA and other CPT I inhibitors probably interact at the catalytic center of the enzyme, not with a regulatory subunit. The amino acid sequences of rat and human CPT II (deduced from cDNA clones) show them to be similar proteins (greater than 80% identity) but encoded by mRNAs of significantly different sizes. Efforts to clone and sequence the cDNA for rat liver CPT I are presently underway.

Amino Acid Sequence↗
PubMed1991

Urban/rural differences in home health patients and services.

Using data from the Wisconsin Annual Survey of Home Health Agencies, we describe urban/rural differences for home health care patients. Our findings indicate that urban dwellers are more likely to be home health patients than are rural residents. Urban home health patients are more apt to be nonelderly, male, and have "other conditions" as their primary diagnosis. They are also likely to be more physically dependent and to receive home care longer. Urban home health patients are more typical of long-term care patients, whereas rural patients may be better described as recipients of postacute care, often recovering from diabetes and heart attacks. Possible problems with rural access to home health care are discussed.

Activities of Daily Living↗
PubMed1991

The financial effect of physician practice style on hospital resource use.

Several specifications of a statistical model were used to measure the effect that internal medicine attending physicians had on inpatient charges and length of stay at a large urban teaching hospital. The study was based on a sample of 1,458 patients discharged during 1985-1987 with 12 common principal diagnosis clusters. The relationship between 31 physicians' clinical decisions and hospital charges and length of stay was analyzed controlling for patients' health status, as measured by demographic characteristics, diagnostic group, and ratings for the Severity of Illness Index (SOII). Results indicated that attending physicians were statistically significant predictors of the log of total charges (p = .0030) and the log of length of stay (p less than .0001), and not as significant predictors of untransformed total charges (p = .1255). Equivalent results were obtained when overall SOII ratings were replaced by SOII subscale ratings for the presenting stage of the principal diagnosis on admission. Examination of individual physician regression coefficients revealed that physicians varied within a 40 percent range of generated per patient charges. No significant differences in mortality, early readmissions or residual impairment on discharge were found between the ten highest and ten lowest resource use physicians. The conservatively estimated range of attending physician practice variations observed in this study has serious financial implications for hospitals operating under incentives to minimize operating costs, particularly for teaching hospitals facing reductions in subsidies for graduate medical education.

Aged↗
PubMed1990

Physical properties of glycosyl diacylglycerols. 2. X-ray diffraction studies of a homologous series of 1,2-Di-O-acyl-3-O-(alpha-D-glucopyranosyl)-sn-glycerols.

X-ray diffraction methods were used to characterize the thermotropic polymorphism exhibited by aqueous dispersions of a homologous series of 1,2-O-acyl-3-O-(alpha-D-glucopyranosyl)-sn-glycerols. Upon cooling from temperatures at which the acyl chains of these lipids are melted, all of these compounds form structures that exhibit both low-angle and wide-angle diffraction patterns consistent with the formation of lamellar L beta gel phases. After a suitable protocol of low-temperature annealing, complex diffraction patterns consistent with the formation of highly ordered, lamellar, crystal-like phases are obtained. These patterns are similar for all of the compounds studied, suggesting that the unit cell structure is invariant. The assumption that the unit cell structure is invariant permits the assignment of phases to the diffraction orders, thereby making possible the construction of electron density profiles. These electron density profiles indicate that the crystal-like phases of these lipids are poorly hydrated structures with the hydrocarbon chains inclined at 35 degrees to the bilayer normal. The diffraction patterns of the crystal-like phases of these lipids changed abruptly at the calorimetrically determined phase transition temperatures to those characteristic of either lamellar liquid crystalline phases (N less than or equal to 17) or inverted nonbilayer phases. With these X-ray diffraction data we demonstrate that, at elevated temperatures, the shorter chain homologues (N less than or equal to 16) form cubic phases of the Pn3m space group, whereas the longer chain compounds form inverted hexagonal phases.

Diglycerides↗
PubMed1990

Cloning, sequencing, and expression of a cDNA encoding rat liver mitochondrial carnitine palmitoyltransferase II.

We report the isolation and characterization of a full-length cDNA encoding rat liver carnitine palmitoyltransferase II (CPT II). Beginning with the purified protein CNBr fragments were generated and sequenced. Corresponding oligonucleotides were used to screen a rat liver cDNA library constructed in the plasmid cloning vector, pcDV. The clone ultimately obtained consisted of a 62 nucleotide 5'-untranslated region, a single open reading frame of 1,974 bases predicting a protein of 658 amino acids (Mr = 74,119), and a 3'-untranslated segment of 260 nucleotides followed by the poly (A) tail. The identity of the cDNA was confirmed by the findings that (a) the open reading frame encoded all three peptides found in the original protein; (b) a fourth peptide synthesized from a portion of the deduced amino acid sequence and used to immunize a rabbit resulted in the generation of an antibody that recognized pure CPT II on a Western blot; (c) in vitro transcription and translation of the cDNA (ligated into pBlue-script KS (+] generated a protein that was specifically immunoprecipitated by anti-CPT II antibody and having a Mr slightly greater than that of mature CPT II; (d) transfection of COS cells with the cDNA subcloned into the expression vector, pCMV4, resulted in a 6-fold induction of mitochondrial CPT II catalytic activity. It seems likely that the de novo synthesized enzyme gains entry into the mitochondrion via a targeting peptide that is subsequently cleaved. The mature protein probably associates (relatively loosely) with the inner membrane through a limited number of membrane spanning domains. The predicted amino acid sequence of CPT II shows strong identity with those of two other acyltransferases, namely, rat liver peroxisomal carnitine octanoyltransferase and porcine choline acetyltransferase.

Acyltransferases↗
PubMed1990

Phorbol esters induce immediate-early genes and activate cardiac gene transcription in neonatal rat myocardial cells.

The mechanisms which transduce intracellular signals for the accumulation of myofibrillar protein during the onset of myocardial cell hypertrophy are unknown. Although previous studies in skeletal muscle cells have suggested that the activation of protein kinase C induces de-differentiation, including the selective disassembly of myofibrils and inhibition of myofibrillar protein synthesis, the present study demonstrates that phorbol esters which activate protein kinase C lead to the accumulation of an individual contractile protein, myosin light chain-2 (MLC-2) and produce several features of myocardial cell hypertrophy. Utilizing immunoblotting and indirect immunocytofluorescence with MLC antisera, the present study demonstrates a several-fold increase in the content of MLC-2, and a marked increase in the assembly of MLC into organized contractile units in individual neonatal rat myocardial cells following treatment with phorbol 12-myristate 13-acetate (PMA). The concentration of PMA required to elicit this response and the lack of a response with an inactive phorbol ester is consistent with the activation of a protein kinase C dependent pathway. Furthermore, PMA treatment results in the rapid induction of a program of immediate-early gene expression (including the c-fos and c-jun proto-oncogenes, and an inducible zinc finger containing gene, egr-l), and activates cardiac gene transcription as assessed by nuclear run-on analyses. The results of the present study suggest the possibility that a protein kinase C dependent pathway may be involved in the up-regulation of myofibrillar protein content and the activation of cardiac gene transcription during growth and hypertrophy of neonatal rat myocardium, and that the induction of a program of immediate-early gene expression may be linked to this response.

Animals↗
PubMed1990

Inhibition of ribonucleotide reduction in CCRF-CEM cells by 2',2'-difluorodeoxycytidine.

The new deoxycytidine analogue 2',2'-difluorodeoxycytidine (dFdC) is a specific inhibitor of DNA synthesis that has marked cytotoxicity and therapeutic activity. A 2-hr incubation with 0.1-10 microM dFdC decreased cellular viability 78-97%. This treatment reduced deoxynucleoside triphosphate pools, similar to the action of the ribonucleotide reductase inhibitor hydroxyurea. The most pronounced decrease occurred in the dCTP pool, quantitatively followed by the decrease of dATP, dGTP, and dTTP. In contrast, inhibition of DNA synthesis by arabinosylcytosine did not affect the dCTP level, whereas dATP, dGTP, and dTTP pools increased, but less than 2-fold. The incorporation of [5-3H]cytidine into the dCTP pool, a measure of ribonucleotide reductase activity in whole cells, was reduced to 3% of controls by 0.1 microM dFdC, but to only 40% by 0.1 microM ara-C. Each drug decreased incorporation of [5-3H]cytidine into DNA to a similar extent (greater than 94%), suggesting limitation by a reaction proximal to this step. The cellular concentration of dFdC 5'-diphosphate was 0.3 microM at 50% inhibition of the in situ activity of ribonucleotide reductase. Direct assays of partially purified ribonucleoside diphosphate reductase (EC 1.17.4.1) demonstrated 50% inhibition by 4 microM dFdC 5'-diphosphate; dFdC 5'-triphosphate was much less inhibitory. We conclude that dFdC 5'-diphosphate acts as an inhibitor of ribonucleoside diphosphate reductase.

Cell Line↗
PubMed1989

Structure, organization, and expression of the rat cardiac myosin light chain-2 gene. Identification of a 250-base pair fragment which confers cardiac-specific expression.

The present study characterized the structure, organization, and expression of the rat cardiac myosin light chain (MLC) -2 gene. The rat cardiac MLC-2 gene has seven exons which display complete conservation with the exon structure of the rat fast twitch skeletal MLC-2 gene. A 250-base pair (bp) sequence of the 5'-flanking region contains CArG motifs and additional cis elements, each greater than 10 bp in length, which were conserved in sequence and relative position with the chick cardiac MLC-2 gene. A series of MLC-2/luciferase fusion genes consisting of nested 5' deletions of the MLC-2 5'-flanking region were constructed and transfected into primary neonatal rat myocardial cells and a non-myocardial cell line (CV-1), demonstrating that this 250 bp of the MLC-2 5'-flanking region was sufficient to confer cardiac specific expression on a luciferase reporter gene. This study suggests the presence of important proximal regulatory sequences in the MLC-2 5'-flanking region which are capable of directing the cardiac specific expression of the rat cardiac myosin light chain-2 gene.

Amino Acid Sequence↗
PubMed1989

Effects of lipid packing on polymorphic phase behavior and membrane properties.

The self assembly of phospholipid molecules in the bilayer form was considered in terms of equivalent molecular shapes representing intermolecular forces. The equivalent size of each phospholipid headgroup was approximated by the net atomic volume plus the volume of the associated water molecules, which was derived from water/hydrocarbon partitioning experiments. The equivalent lengths of unsaturated acyl chains were derived from the retention time data from chromatographic measurements. The spontaneous curvature of various phospholipid monolayers was calculated from their equivalent molecular shapes, and the energy required to flatten them to the bilayer plane was calculated, using the known bending modulus. With increasing bending energy, the mixtures showed increasing susceptibility to phospholipase A2, facilitated lipid transfer rate by phospholipid exchange proteins, permeability to carboxyfluorescein, incorporation of human erythrocyte proteins, and calcium transport by Ca-ATPase from sarcoplasmic reticulum in reconstituted vesicles. When the calculation was applied to known lipid compositions of nine cellular membranes, the protein/lipid ratio and phospholipid/cholesterol ratio were found to have a positive and a negative correlation, respectively, with the latent bending energy of the phospholipids. The energy expense in conforming to a bilayer phase may be an important physical parameter regarding the activity and the biogenesis of membranes.

Cholesterol↗