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Biomedical subjects

S Datta

Publications and source records attributed to S Datta.

At least 289 records · Page 16Linked to original sources
PubMed1987

Structure and expression of dog apolipoprotein C-II and C-III mRNAs. Implications for the evolution and functional constraints of apolipoprotein structure.

The complete cDNAs for canine apolipoprotein (apo) C-II and C-III have been cloned in the plasmid vector, pUC-18, and the phage vector, lambda gt11. With 32P-labeled nick-translated cDNAs as hybridization probes, apoC-II and apoC-III mRNAs were found to be expressed in canine liver, jejunum, ileum, colon, pancreas, spleen, ovary, and bladder at the following relative concentrations: 100, 35, 30, 10, 15, 5, 30, and 2% for apoC-II, and 100, 35, 15, 2, 20, 5, 30, and 5% for apoC-III. Neither mRNA was detected in canine brain, kidney, lung, muscle, oviduct, uterus, or testis. Both mRNAs are approximately 700 nucleotides in length. Complete sequence analysis revealed that apoC-II mRNA codes for a protein of 101 amino acid residues including 22 residues in the signal peptide and 79 residues in the mature peptide region. ApoC-III mRNA codes for a protein of 100 amino acid residues, including 20 in the signal peptide and 80 residues in the mature peptide region. It contains a single cysteine residue in position 54. Secondary structural analysis shows the presence of two regions each of amphipathic helix for apoC-II (residues 14-33 and 44-63) and apoC-III (residues 16-33 and 50-69). Computer analysis of the amphipathic helical regions demonstrates the importance of variation in the dihedral angles of the side chains in the charge microenvironment of the helices. Statistical analysis of sequence similarities between human apoC-II, apoC-III, apoA-II, apoA-I, apoE, and apoA-IV, and dog apoC-II and apoC-III, and rat apoC-III, apoA-II, apoA-I, apoE, and apoA-IV indicates that all these proteins have evolved rapidly, especially in the rat in which apoC-III has evolved at three times the rate in man and dog. Further, the shorter apolipoproteins, apoA-II, apoC-II, and a-poC-III, have evolved more rapidly than the longer ones, apoA-I, apoE, and apoA-IV. Some regions in the individual apolipoproteins are better conserved than others. The rate of evolution of individual regions seems to be related to the stringency of their functional requirements.

Amino Acid Sequence↗
PubMed1987

Elective postoperative external radiotherapy after hysterectomy in early-stage carcinoma of the cervix. Is additional vaginal cuff irradiation necessary?

This retrospective study of 44 patients who received postoperative radiotherapy soon after hysterectomy for early Stage IB to IIA carcinoma of the cervix was undertaken to determine the need for additional vaginal cuff irradiation (AVCI) following external pelvic irradiation (EPI). The reasons for adjuvant radiotherapy administration were mostly the nonradical type of performed hysterectomy or the presence of adverse prognostic histopathologic features. The overall survival and local control rates were 80% (35/44) and 89% (39/44) at 2 years and 63% (26/41) and 80% (33/41) at 5 years, respectively. Although the 2-year results seemed to indicate that AVCI was necessary, this was not supported by the obtained longer term data. Moreover, it appeared that AVCI could perhaps be safely done after EPI, when occult early-stage cervical carcinoma was found after total abdominal hysterectomy. Data from this study and the literature concerning AVCI's appropriate role in this particular disease condition are discussed.

Carcinoma↗
PubMed1987

Identification of the sequences controlling cyclic AMP regulation and cell-type-specific expression of a prestalk-specific gene in Dictyostelium discoideum.

We have cloned and analyzed a developmentally and spatially regulated prestalk cell-specific gene from Dictyostelium discoideum. The gene encodes a protein highly homologous to the lysosomal cysteine proteinases cathepsin H and cathepsin B. Amino acid comparisons between these enzymes showed that the active-site amino acids were conserved, as were amino acids known to be important for catalysis and residues which form the intramolecular cysteine bridges. We have constructed a series of internal deletions, duplications, and linker scanner mutations within the region 300 base pairs 5' to the cap site. Analysis of expression of the mutations in transformants identified a approximately 35-base pair GC-rich region containing a dAdC/dGdT palindromic repeat and a G-rich box which is homologous to the 3' GT half of the palindromic repeat. Deletion or disruption of the G box resulted in a approximately 50-fold drop in the level of expression of the gene fusion in transformants in response to cyclic AMP in single-cell culture but did not affect the temporal pattern of regulation or control by cyclic AMP. The expression of such constructs during normal multicellular differentiation paralleled that of the endogenous gene; however, the level of RNA from the constructs was only approximately 10-fold lower than that of constructs containing the G box. Deletion of the 3' half of the palindromic sequence and the G box region resulted in a dramatic decrease in the level of transcription, although the constructs still showed proper temporal expression. These results suggest that this 35-base-pair region acts as an important part of the regulatory region for cell type and cyclic AMP regulation.

Amino Acid Sequence↗
PubMed1987

Effects of fentanyl and sufentanil on peripheral mammalian nerves.

The effects of fentanyl and sufentanil on peripheral nerves were evaluated in isolated sheathed and desheathed rabbit vagus nerves. The action potential amplitudes of A and C fibers were recorded before and after a 30-min exposure to 50 and 100 micrograms/ml of fentanyl and sufentanil. A reversible decrease in the action potential amplitude of A fibers in desheathed nerves was observed after exposure to 100 micrograms/ml of each drug. The action potential amplitude of C fibers was also decreased but not to the same degree as was the A fiber action potential. Pretreatment with naloxone failed to block the reduction in action potential amplitude produced by the two opiates. No evidence of irreversible conduction blockade indicative of local neural toxicity was seen in these studies. The results suggest that high concentrations of fentanyl and sufentanil may exert a weak local anesthetic-type action on peripheral nerves.

Action Potentials↗
PubMed1987

Effect of pregnancy on bupivacaine-induced conduction blockade in the isolated rabbit vagus nerve.

Bupivacaine-induced conduction blockade of A, B, and C fibers of the isolated vagus nerve was compared in fourteen pregnant and fourteen nonpregnant rabbits. After a control period in HEPES-Liley solution, the isolated nerves were exposed to bupivacaine concentrations of 0.1 mM to 1.0 mM. After 30 min exposure, the nerves were stimulated supramaximally and the percent reduction in amplitude of A, B, and C fiber compound action potentials was recorded. Linear regressions were fitted by the least squares method. The A fiber conduction blockade was consistently greater in the nerves from pregnant rabbits (P less than 0.001). The slope of the C fiber dose-response curves was also significantly greater in nerves from pregnant rabbits (P less than 0.01). The results indicate that the response of isolated nerves from pregnant animals to local anesthetic-induced conduction blockade differs from that of nerves from nonpregnant animals. However, it is not certain whether the difference is related simply to a more rapid diffusion and shorter onset of block or an enhanced sensitivity of the nerve membrane during pregnancy.

Anesthesia, Epidural↗
PubMed1986

Salt-dependent changes in the DNA binding co-operativity of Escherichia coli single strand binding protein.

The co-operative nature of the binding of the Escherichia coli single strand binding protein (SSB) to single-stranded nucleic acids has been examined over a range of salt concentrations (NaCl and MgCl2) to determine if different degrees of binding co-operativity are associated with the two SSB binding modes that have been identified recently. Quantitative estimates of the binding properties, including the co-operativity parameter, omega, of SSB to single-stranded DNA and RNA homopolynucleotides have been obtained from equilibrium binding isotherms, at high salt (greater than or equal to 0.2 M-NaCl), by monitoring the fluorescence quenching of the SSB upon binding. Under these high salt conditions, where only the high site size SSB binding mode exists (65 +/- 5 nucleotides per tetramer), we find only moderate co-operativity for SSB binding to both DNA and RNA, (omega = 50 +/- 10), independent of the concentration of salt. This value for omega is much lower than most previous estimates. At lower concentrations of NaCl, where the low site size SSB binding mode (33 +/- 3 nucleotides/tetramer) exists, but where SSB affinity for single-stranded DNA is too high to estimate co-operativity from classical binding isotherms, we have used an agarose gel electrophoresis technique to qualitatively examine SSB co-operativity with single-stranded (ss) M13 phage DNA. The apparent binding co-operativity increases dramatically below 0.20 M-NaCl, as judged by the extremely non-random distribution of SSB among the ssM13 DNA population at low SSB to DNA ratios. However, the highly co-operative complexes are not at equilibrium at low SSB/DNA binding densities, but are formed only transiently when SSB and ssDNA are directly mixed at low concentrations of NaCl. The conversions of these metastable, highly co-operative SSB-ssDNA complexes to their equilibrium, low co-operativity form is very slow at low concentrations of NaCl. At equilibrium, the SSB-ssDNA complexes seem to possess the same low degree of co-operativity (omega = 50 +/- 10) under all conditions tested. However, the highly co-operative mode of SSB binding, although metastable, may be important during non-equilibrium processes such as DNA replication. The possible relation between the two SSB binding modes, which differ in site size by a factor of two, and the high and low co-operativity complexes, which we report here, is discussed.

Allosteric Regulation↗
PubMed1986

Alpha adrenergic system in medial preoptic area involved in sleep-wakefulness in rats.

The study is aimed at investigating the possible involvement of adrenergic mechanisms in the medial preoptic area (mPOA) for modulation of sleep-wakefulness in rats. In this study, saline, norepinephrine (NE), phenoxybenzamine (PBZ) and propranolol (PROP) were injected in the mPOA in different groups of male rats during the day and night. NE and PBZ were injected, during the day and the night respectively, in some control areas adjoining the mPOA in two other groups of animals. Arousal was produced by NE, and sleep by PBZ when they were applied in the mPOA. All other procedures, including application of NE and PBZ in the control areas and beta blocker (PROP) in the mPOA, did not produce alterations in sleep-wakefulness. These findings provide support for a physiological role played by the alpha adrenergic system in the mPOA for arousal, and area specificity of action of this system.

Animals↗
PubMed1986

Cellular and subcellular distribution of a cAMP-regulated prestalk protein and prespore protein in Dictyostelium discoideum: a study on the ontogeny of prestalk and prespore cells.

We have analyzed a developmentally and spatially regulated prestalk-specific gene and a prespore-specific gene from Dictyostelium. The prestalk gene, pst-cathepsin, encodes a protein highly homologous to the lysosomal cysteine proteinases cathepsin H and cathepsin B. The prespore gene encodes a protein with some homology to the anti-bacterial toxin crambin and has been designated beejin. Using the lambda gtll system, we have made polyclonal antibodies directed against a portion of the protein encoded by pst-cathepsin and other antibodies directed against the beejin protein. Both antibodies stain single bands on Western blots. By immunofluorescence and Western blots, pst-cathepsin is not present in vegetative cells or developing cells during the first approximately 10 h of development. It then appears with a punctate distribution in a subset of developing cells. Beejin is detected only after approximately 15 h of development, also in a subset of cells. Pst-cathepsin is distributed in the anterior approximately 1/10 of migrating slugs and on the peripheral posterior surfaces of slugs. Beejin is distributed in the posterior region of slugs. Expression of both pst-cathepsin and beejin can be induced in subsets of isolated cultured cells by a combination of conditioned medium and extracellular cAMP in agreement with the regulation of the mRNAs encoding these proteins. We have used the antibodies as markers for cell type to examine the ontogeny and the spatial distribution of prestalk and prespore cells throughout multicellular development. Our findings suggest that prestalk cell differentiation is independent of position within the aggregate and that the spatial localization of prestalk cells within the multicellular aggregate arises from sorting of the prestalk cells after their induction. We have also found a class of cell in developing aggregates that contains neither the prestalk nor the prespore markers.

Antibodies, Monoclonal↗
PubMed1986

Spatial and temporal regulation of a foreign gene by a prestalk-specific promoter in transformed Dictyostelium discoideum.

We analyzed a developmentally regulated prestalk-specific gene from Dictyostelium discoideum encoding a cathepsin-like protease. A hybrid gene was constructed by fusing 2.5 kilobases of 5' flanking sequences and part of the coding region of the gene in-frame to the Escherichia coli beta-glucuronidase gene and was transformed into D. discoideum cells. In cells transformed with this vector, the gene fusion showed the same temporal regulation as the endogenous gene during multicellular development and, like endogenous prestalk genes, was highly inducible by cyclic AMP in in vitro cell cultures. Moreover, immunofluorescence studies showed that the fusion protein had the same spatial distribution within the migrating pseudoplasmodium as the endogenous gene. The results indicate that the regions of the D. discoideum prestalk-specific cathepsin gene contain all the necessary information for proper temporal, spatial, and cyclic AMP regulation of a prestalk cell-type gene in D. discoideum transformants and leads the way for experiments to identify the cell-type-specific regulatory elements.

DNA Restriction Enzymes↗