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S C Martin

Publications and source records attributed to S C Martin.

65 records · Page 4Linked to original sources

In vitro phosphorylation of serum albumin by two protein kinases: a potential pitfall in protein phosphorylation reactions.

Bovine albumin was phosphorylated by both cAMP-dependent protein kinase and casein kinase I to a significant extent. Other albumins were also tested and it was found that the extent of phosphorylation varied with the species of origin of the albumin, but was between 1 and 3 mol phosphate per mole albumin for the cAMP-dependent protein kinase-catalyzed reactions. The phosphorylation occurred at and above pH 7.5 and required the presence of thiol reagents. Phosphoamino acid analyses of bovine albumin showed that it was phosphorylated on at least two serine residues. The phosphorylation could not be demonstrated in vivo.

Animals↗

Deinduction of transcription of Xenopus 74-kDa albumin genes and destabilization of mRNA by estrogen in vivo and in hepatocyte cultures.

The goal of this study is to explain the molecular basis of the marked deinduction of Xenopus albumin synthesis and secretion accompanying the activation of vitellogenin genes by estrogen. We have characterized by restriction analysis, DNA sequencing and hybrid-selected translation of mRNA, a cloned cDNA specifying the two 74-kDa albumins which constitute the predominant circulating form of albumin in Xenopus laevis. Using this recombinant DNA plasmid as a hybridization probe, we have determined the steady-state levels of albumin mRNA, the rate of transcription of the two 74-kDa albumin genes and the stability of the mRNA in male and female Xenopus hepatocytes in vivo and in primary cell cultures following estrogen treatment. In both whole liver and cultured hepatocytes estradiol caused a rapid drop in the steady-state levels of 74-kDa albumin mRNAs, which was reversed spontaneously in the continued presence of the hormone. The concentration of albumin mRNA was substantially higher in male than in female hepatocytes, the hormonal effect being more marked in male than in female hepatocytes. The decrease in steady-state levels of mRNA was anticipated in male hepatocytes by a 70% inhibition of rate of transcription of albumin genes within 2 h of exposure to estradiol, as measured by run-off transcription in liver nuclei isolated from animals treated in vivo or by determining the absolute transcription rate in cell cultures. In the latter the diminished transcription rate returned to normal within 12 h in the continued presence of the hormone. Estradiol caused a threefold destabilization of albumin mRNA in both male and female hepatocyte cultures to t 1/2 = 3 h and 2 h respectively. The combined effects on rate of or transcription and mRNA stability largely explain the changes in the steady-state levels of mRNA caused by hormone administration. Comparison of the kinetics of transcription rates of vitellogenin and albumin genes in vivo and in vitro reveals a striking reciprocity in the selective activation of the inducible genes and deinduction of the constitutively expressed genes at the early stages of response of Xenopus hepatocytes to estrogen.

Albumins↗

Haematological and biochemical values in horses naturally infected with Strongylus vulgaris.

The concentrations of serum proteins (beta 1, beta 2, gamma, alpha 1, alpha 2 globulins and albumin) and absolute numbers of eosinophils, neutrophils and lymphocytes were examined in 64 naturally infected horses and ponies in which the number of larvae of Strongylus vulgaris in the cranial mesenteric artery and the severity of the lesion of verminous arteritis could be determined. The horses were grouped according to the number of larvae found and the severity of the arteritis. The results demonstrated that, although some significant deviation from a random distribution occurred in certain of the values (chi 2 test), there was considerable individual variation in the values obtained for individual animals within groups and overlap of the range of values between groups. Also the number of larvae present in the artery did not necessarily accurately reflect the severity of the arterial lesion. Thus, the parameters examined could not be used reliably to estimate the intensity of infection with S vulgaris in an individual animal.

Animals↗

Two forms of poliovirus VPg result from amino acid modification of a single viral protein.

The protein (VPg) covalently attached to the 5' terminus of poliovirus RNA has been reported to resolve into two forms, separable by electrofocusing, yet the viral gene sequence predicts only one apparent gene for VPg. The two VPg species were separated and analyzed for structural differences. The protein contains no phosphorylated amino acid residues other than the junction tyrosine linked to the nucleic acid moiety. After isolation, the acidic form of VPg remains stable, but the more basic form again generates a distribution of both species. This suggests some lability or modification of at least one amino acid residue postsynthesis. The position of the alteration in the protein was localized to the amino-terminal tryptic peptide, containing nine amino acids.

Amino Acid Sequence↗

Structure of poliovirus replicative intermediate RNA. Electron microscope analysis of RNA cross-linked in vivo with psoralen derivative.

The structure of the poliovirus replicative intermediate RNA was examined by electron microscopy after cross-linking in vivo with 4'-aminomethyl-4,5',8-trimethylpsoralen. After purification from infected cells, undenatured RI appeared as a double-stranded backbone of genome length, with an average of three (and occasionally up to eight) nascent, single-stranded tails. After denaturation, however, only single strands of heterogeneous length were visualized, indicating that the RI in the cell contains little or no duplex structure, and thus nascent chains are only transiently hydrogen-bonded to their template over short regions. The double-stranded backbone of undenatured RI, observed previously by others and in these experiments, is due to collapse of complementary chains during the deproteinization and purification procedures. The effectiveness of the in vivo cross-linking procedure was demonstrated by the complete inhibition of viral RNA synthesis in treated cells and by direct binding of [3H]AMT to RI molecules in vivo. Mature polio virions are impermeable to AMT; however, growth of virus in cells incubated with AMT in the dark resulted in normal yields of virus particles containing RNA genomes, whose infectivity could be subsequently photo-inactivated. The frequency of AMT-induced cross-linking was determined by analyses of double-stranded poliovirus RNA (RF). Cross-linking in vitro followed by spreading for electron microscopy under denaturing conditions yielded bubbled duplex structures with a minimum of one interstrand cross-link per 80 base-pairs. RF cross-linked in vivo also showed extensive cross-linking, decreased about fivefold from the in vitro cross-linked value. Thus, the failure to detect cross-linked RI under these conditions indicates that extensive base-pairing does not exist in vivo.

Base Composition↗

In vitro induction of lymphocyte responsiveness by a Strongylus vulgaris-derived mitogen.

Proliferation in vitro of peripheral blood lymphocytes both from horses infected with Strongylus vulgaris and from helminth-free ponies was observed in the presence of extracts of the fourth and fifth stage larvae and adults of S. vulgaris. In addition, S. vulgaris extracts induced transformation in cultures of peripheral blood lymphocytes from sheep and dogs and in mouse spleen cell cultures. Nylon wool non-adherent, T cell enriched fractions of lymphocytes from both mice and horses were stimulated by the S. vulgaris larval mitogen while no proliferation was observed in cultures containing nylon wool adherent, B cell enriched fractions. Macrophage co-operation appeared not to be necessary for S. vulgaris mitogen-induced transformation of spleen cells. The S. vulgaris mitogen stimulated a subpopulation of mouse spleen cells different from those responsive to PHA, Con A and LPS. These cells might be T helper cells since B cells were stimulated to proliferate in the presence of both T cells and S. vulgaris larval mitogen. In addition, the supernatant of in vitro cultured larvae of S. vulgaris induced slight, but significant transformation of equine peripheral blood lymphocytes. Therefore, it is possible that the S. vulgaris mitogen released by both viable parasites and degenerating larvae might induce T cell dependent production of immunoglobulin in vivo and account for the beta-globulinaemia, of which IgG(T) is a major component, in S vulgaris infected horses.

Animals↗

Drosophila mitochondrial DNA: a novel gene order.

Part of the replication origin-containing A+T-rich region of the Drosophila yakuba mtDNA molecule and segments on either side of this region have been sequenced, and the genes within them identified. The data confirm that the small and large rRNA genes lie in tandem adjacent to that side of the A+T-rich region which is replicated first, and establish that a tRNAval gene lies between the two rRNA genes and that URF1 follows the large rRNA gene. The data further establish that the genes for tRNAile, tRNAgln, tRNAf-met and URF2 lie in the order given, on the opposite side of the A+T-rich region to the rRNA genes and, except for tRNAgln, are contained in the opposite strand to the rRNA, tRNAval and URF1 genes. This is in contrast to mammalian mtDNAs where all of these genes are located on the side of the replication origin which is replicated last, within the order tRNAphe, small (12S) rRNA, tRNAval, large (16S) rRNA, tRNAleu, URF1, tRNAile, tRNAgln, tRNAf-met and URF2, and, except tRNAgln, are all contained in the same (H) strand. In D. yakuba URF1 and URF2, the triplet AGA appears to specify an amino acid, which is again different from the situation found in mammalian mtDNAs, where AGA is used only as a rare termination codon.

Amino Acid Sequence↗

A multidimensional measure of fatigue for use with cancer patients.

PURPOSE: The purpose of this study was to develop and validate a multidimensional measure of fatigue for use with cancer patients. DESCRIPTION OF STUDY: Items for the Multidimensional Fatigue Symptom Inventory (MFSI) were generated through literature review, discussion with healthcare providers, and a survey of currently available measures of fatigue. The 83-item MFSI was designed to assess global, somatic, affective, cognitive, and behavioral symptoms of fatigue. The instrument was administered on three occasions to 275 women who had received or were undergoing treatment for breast cancer and 70 women with no history of cancer. Reliability, validity, and factorial structure of the MFSI were analyzed. RESULTS: The factor analysis produced five empirically derived scales that correspond generally to the five rationally derived scales. Reliability of the rationally and empirically derived scales was excellent (alpha coefficients .87 to .96). Estimates of test-retest reliability were also favorable. Other results support the validity of both the rationally and empirically derived scales. The MFSI appears to be sensitive to fatigue, accurately discriminating cancer patients from control subjects and between patients with varying levels of performance status. CLINICAL IMPLICATIONS: The MFSI may be useful in identifying patterns of fatigue within individual patients and across treatment modalities. Such specificity may allow the clinician to develop, implement, and evaluate interventions that are targeted for differing patterns of fatigue. Because the measure is keyed to a 1-week time frame, it may be useful during the course of cancer treatment. The MFSI appears to be a valid and reliable tool to assess the full spectrum of symptoms that characterize the construct of fatigue.

Adult↗