Biomedical subjects

G R Martin

Publications and source records attributed to G R Martin.

At least 271 records · Page 15Linked to original sources

PubMed1986

The classification of peripheral 5-HT2-like receptors using tryptamine agonist and antagonist analogues.

In a previous study, we attempted to verify the classification of 5-hydroxytryptamine2 (5-HT2) receptors in three vascular tissues, by use of the conventional antagonists, ketanserin, spiperone, methysergide and trazodone. However, it was not possible to conclude homogeneity of the receptor type in the three tissues due to the inconsistent behaviour of these antagonists, in particular, their apparently variable affinities between the tissues. These results led to the reliability of the conventional antagonists being questioned as receptor probes. In the present study, a set of tryptamine analogues were investigated in two of the tissues, the rabbit aorta and the rat jugular vein. Unlike the conventional antagonists, these compounds bear a close chemical relation to the natural agonist, 5-HT. In both tissues, alpha, alpha-dimethyltryptamine demonstrated apparently simple competitive antagonism of 5-HT-induced constrictions. Its affinity was estimated to be the same in each case. The affinities and relative efficacies of 5-HT, 5-cyanotryptamine, N,N-dimethyltryptamine and N-benzyl-5-methoxytryptamine were also found to be indistinguishable between the two tissues. Unlike the conventional 5-HT2 receptor antagonists, these tryptamine analogues failed to distinguish between the 5-HT receptors in the rabbit aorta and rat jugular vein implying that they truly belong to the same class. In view of this result, it is suggested that simple tryptamine analogues are more reliable probes for 5-HT receptor classification than ligands which bear little or no chemical relation to the natural agonist.

Animals↗

PubMed1986

Routine evaluation of the nonradiometric BACTEC NR 660 system.

The present study compares the sensitivity, accuracy, and practicability of a nonradiometric blood culture system with those of the standard blood culture system in use in our laboratory. A total of 1,080 sets of four blood cultures bottles were compared, giving 143 positive aerobic-medium associations and 139 positive anaerobic-medium associations. The conventional system recovered 171 isolates, whereas the BACTEC NR 660 system (Johnston Laboratories, Towson, Md.) recovered 155 isolated. There were no significant differences in total isolates with either system (P more than 0.05). The BACTEC NR 660 system recovered a slightly lower number of Serratia marcescens and Streptococcus faecalis but a higher number of Pneumococcus spp. The mean detection times were 55.9 h with the conventional system and 31.6 h with the BACTEC NR 660 system (P less than 0.001). Our results demonstrate that the BACTEC NR 660 system is suitable for routine use.

Bacteria, Aerobic↗

PubMed1986

Methylation of the mouse hprt gene differs on the active and inactive X chromosomes.

It has been proposed that DNA methylation is involved in the mechanism of X inactivation, the process by which equivalence of levels of X-linked gene products is achieved in female (XX) and male (XY) mammals. In this study, Southern blots of female and male DNA digested with methylation-sensitive restriction endonucleases and hybridized to various portions of the cloned mouse hprt gene were compared, and sites within the mouse hprt gene were identified that are differentially methylated in female and male cells. The extent to which these sites are methylated when carried on the active and inactive X chromosomes was directly determined in a similar analysis of DNA from clonal cell lines established from a female embryo derived from a mating of two species of mouse, Mus musculus and Mus caroli. The results revealed two regions of differential methylation in the mouse hprt gene. One region, in the first intron of the gene, includes four sites that are completely unmethylated when carried on the active X and extensively methylated when carried on the inactive X. These same sites are extensively demethylated in hprt genes reactivated either spontaneously or after 5-azacytidine treatment. The second region includes several sites in the 3' 20kilobases of the gene extending from exon 3 to exon 9 that show the converse pattern; i.e., they are completely methylated when carried on the active X and completely unmethylated when carried on the inactive X. At least one of these sites does not become methylated after reactivation of the gene. The results of this study, together with the results of previous studies by others of the human hprt gene, indicate that these regions of differential methylation on the active and inactive X are conserved between mammalian species. Furthermore, the data described here are consistent with the idea that at least the sites in the 5' region of the gene play a role in the X inactivation phenomenon and regulation of expression of the mouse hprt gene.

Animals↗

PubMed1986

Spontaneous expression of the c-sis gene and release of a platelet-derived growth factorlike molecule by human alveolar macrophages.

Alveolar macrophages from normal individuals and patients with interstitial lung diseases spontaneously expressed a 4.2-kilobase mRNA complementary to the c-sis gene, a proto-oncogene coding for one of the chains of platelet-derived growth factor (PDGF). Concomitantly, these cells released a mediator with the properties of PDGF, including: chemotactic factor for smooth muscle cells whose activity was resistant to heat and acid, but sensitive to reduction; mitogenic (competence) activity for fibroblasts; ability to compete with PDGF for its receptor; and precipitated by an anti-PDGF antibody. While blood monocytes did not contain c-sis mRNA transcripts, monocytes matured in vitro expressed c-sis, consistent with the concept that expression of c-sis occurs during the differentiation of monocytes into alveolar macrophages. Together with the known actions of PDGF, these observations suggest that the c-sis proto-oncogene and its PDGF product are part of the armamentarium available to the alveolar macrophages for normal lung defense and participation in lung inflammation.

Chemotaxis↗

PubMed1986

Distribution of basement membrane antigens in glomeruli of mice with autoimmune glomerulonephritis.

Glomerulonephritis was induced in mice by the repeated injection of human glomeruli or purified glomerular basement membrane. The glomerular basement membranes of nephritic animals were observed to develop subepithelial extensions, "spikes." Although normally Type IV collagen is found throughout the full thickness of basement membranes, the "spikes" reacted with antibodies to laminin but not with antibodies to Type IV collagen. It is proposed that in murine autoimmune glomerulonephritis, the visceral epithelial cells produce an excess of laminin.

Animals↗

PubMed1986

Basement membrane and the invasive activity of metastatic tumor cells.

Many, but not all, metastatic tumor cells have a similar phenotype. These are transformed cells with a high affinity for basement membranes and the ability to produce basement membrane degrading enzymes. Such characteristics help the cells traverse this critical barrier. Traversal may be a rather rare event unless the cells respond to local factors that amplify the numbers of cells recruited to the site and induce the cells to invade. These factors may include tissue-specific attractants and matrix molecules such as laminin. Understanding the specific steps involved in the invasion process should allow development of antimetastatic regimens directed at those activities specific to metastatic tumor cells. Due to the heterogeneity of tumor cells, other mechanisms for metastasis undoubtably exist.

Animals↗

PubMed1985

Isolation and partial characterization of genomic clones coding for a human pro-alpha 1 (II) collagen chain and demonstration of restriction fragment length polymorphism at the 3' end of the gene.

We have isolated two clones containing 19 kilobases (kb) of the human gene coding for a pro-alpha 1 (II) collagen chain from human lambda genomic DNA libraries. A 3' clone, HC2A, was selected by cross-hybridization with a cDNA clone containing sequences coding for the carboxy propeptide of chick type II procollagen. A second clone, HC2B, was obtained by screening the library with the 5' part of HC2A. The sequence analysis of exon 3 corresponding to the C propeptide reveals the presence of stretches of conserved nucleotides between the human and the chick type II procollagen genes. On Northern blots, the human collagen clone hybridizes strongly to a 5.5-kb RNA for the rat type II procollagen chain. Finally, studies of genomic DNAs from normal individuals reveal the presence of a HindIII and a BamHI polymorphic site at the 3' end of the gene.

Amino Acid Sequence↗

PubMed1985

Regulation of type I collagen synthesis. Total pro alpha 1(I) and pro alpha 2(I) mRNAs are maintained in a 2:1 ratio under varying rates of collagen synthesis.

The type I collagen molecule contains two alpha 1(I) chains and one alpha 2(I) chain. Previous investigations, using embryonic chick calvaria, have indicated that the two chains are synthesized in a 2:1 ratio which is controlled at a pretranslational level, since the cells contain twice as much translatable pro alpha 1(I) mRNA as pro alpha 2(I) mRNA. The present report describes hybridization analyses of the cellular levels of total cellular RNAs coding for the pro alpha 1(I) and pro alpha 2(I) chains, using as probes two cloned cDNAs complementary to chick pro alpha 1(I) and pro alpha 2(I) mRNA, respectively. Total cellular RNA was extracted from embryonic chick calvaria, pro alpha 1(I) and pro alpha 2(I) RNA sequences were quantified by Northern hybridization using conditions ensuring that hybridization efficiency and specific radioactivity were the same for the two probes. Similar analyses were carried out on RNA extracted from calvaria with different levels of collagen synthesis after culture in the presence or absence of ascorbic acid. The results for all samples analyzed indicate that total cellular pro alpha 1(I) and pro alpha 2(I) mRNAs are present in a 2:1 ratio which is maintained even during variations in collagen synthesis rate. There is no evidence for regulation mediated by different rates of processing of mRNA precursors, although preferential degradation of the pro alpha 2(I) gene transcript cannot be excluded. Thus, the synthesis of type I procollagen chains is presumably coordinated by transcriptional control.

Animals↗

PubMed1985

Isolation of a teratocarcinoma stem cell lectin implicated in intercellular adhesion.

We have previously identified a cell surface teratocarcinoma stem cell lectin with a fucan/mannan specificity. We now report the purification of the hemagglutinin (lectin) from stem cell conditioned medium by exclusion on a Sepharose 2B column, followed by elution with 0.5M NaCl from DEAE-cellulose, providing an overall purification of about 90-fold. When this material was analyzed, by SDS-polyacrylamide gel electrophoresis, a major band of Mr 56000 was consistently observed. Hemagglutination activity was renatured from the gels and localized exclusively to a region of the gel that, as detected by fluorography, contains only the 56-kDa component. This suggested that this polypeptide comprises the lectin.

Animals↗

PubMed1985

The basement-membrane-like matrix of the mouse EHS tumor: II. Immunohistochemical quantitation of six of its components.

The presence of six substances--laminin, type IV collagen, heparan sulfate proteoglycan, entactin, fibronectin, and the amyloid P component--was investigated immunohistochemically in the matrix of the Engelbreth-Holm-Swarm (EHS) mouse tumor after it had been fixed in formaldehyde (with or without a brief preliminary glutaraldehyde fixation), embedded in Lowicryl K4M, and sectioned for processing through the protein A-gold sequence. Enumeration of the number of gold particles per square micrometer of matrix sections demonstrated that the six substances were present in distinct amounts. The results for each substance were fairly consistent throughout the matrix in three experiments. Furthermore, the available evidence indicated that, with the exception of the amyloid P component, the substances were associated with the cord network of the tumor matrix. Finally, the use of a reconstituted basement membrane containing known amounts of laminin, type IV collagen, and heparan sulfate proteoglycan as a standard, led to the conclusion that, in the tumor matrix, the relative content of laminin to type IV collagen to the proteoglycan was in a ratio of 1:0.6:0.03, suggesting molar ratios of approximately 1:1:0.2, respectively.

Animals↗

PubMed1985

Biological activities of laminin.

Laminin is a multifunctional protein with diverse biological activities. Like fibronectin, it can influence cell adhesion, growth, morphology, differentiation, migration, and agglutination as well as the assembly of the extracellular matrix. Laminin primarily affects cells of epithelial origin, and the response varies depending on the cell. Because most differentiated cells are difficult to maintain in culture, laminin may be an important supplement in studies on cell differentiation in vitro.

Animals↗

PubMed1985

Binding of antibodies in liposomes to extracellular matrix antigens.

We have incorporated antibodies against fibronectin or laminin into liposomes and studied their interaction with insoluble forms of these antigens. The antibodies, after modification by palmitoylchloride, were incorporated into the lipid bilayer by the cholate dialysis method. The antibodies in the liposomes recognized their specific antigen with little reaction to the alternative attachment protein or to albumin (less than 2%). The binding of antibody-containing liposomes to insoluble antigen was inhibited by soluble antibodies to the respective antigens but not by antibodies to other antigens. The affinity constant of the liposome-antibody complex with the antigen was estimated at 1-10 X 10(-9) M liposomes. Thus, antibodies in liposomes retain their reactivity and specificity, and the reaction constant is comparable to that observed for immune complexes.

Animals↗

PubMed1985

Chondronectin: physical and chemical properties.

Chondronectin, the chondrocyte attachment factor, was purified from chicken serum and characterized as to its physical and chemical properties. From sedimentation equilibrium data it was found to have a native molecular weight of 175,800 +/- 800 and a subunit molecular weight of 55,540 +/- 800 in the presence of guanidinium chloride and cysteine, suggesting a trimeric structure linked by disulfide bonds. As visualized by electron microscopy after rotary shadowing, the protein appears compact and globular. The amino acid and carbohydrate compositions of chondronectin are distinct from fibronectin, the fibroblast attachment factor, and laminin, the epithelial cell attachment factor. The activity of chondronectin in promoting attachment of chondrocytes is stable to digestion by collagenase, elastase, and neuraminidase, but is destroyed by trypsin treatment. The data suggest that chondronectin is structurally and chemically distinct from fibronectin and laminin.

Amino Acids↗

PubMed1985

Expression of homologous homeo-box-containing genes in differentiated human teratocarcinoma cells and mouse embryos.

Human chromosome 17 contains a cluster of at least three distinct homeo box regions, Hu1, Hu2, and Hu5, within a 20 kb stretch of DNA. A mouse homeo box region, Mu1, which maps to chromosome 11, was isolated and found to contain extensive nucleotide homology with a 4 kb region encompassing the Hu1 homeo box. The conservation of these chromosomal regions between man and mouse was confirmed by nucleotide sequence analysis: approximately 380 bp of DNA are more than 90% homologous and the 61 amino acids of the homeo box domain are perfectly conserved. We found that a human teratocarcinoma cell line expresses high levels of Hu1 homeo-box-containing mRNA only after differentiation of the cells following treatment with retinoic acid. In contrast, mouse teratocarcinoma cells did not express Mu1 homeo-box-containing mRNA at any stage of differentiation, whereas the expression of such transcripts was detected in mouse embryos from 10 to 17 days of gestation.

Amino Acid Sequence↗

PubMed1985

Expression during embryogenesis of a mouse gene with sequence homology to the Drosophila engrailed gene.

Regions of the mouse and human genomes with strong homology to the Drosophila engrailed gene have been identified by Southern blot analysis. One mouse engrailed-like region, Mo-en.1, has been cloned and partially sequenced; homology with the engrailed gene is localized to a 180 bp engrailed-like homeo box and 63 nucleotides immediately 3' to it. The protein sequence this region can encode includes 81 amino acids, of which 60 (75%) are identical with those of the putative translation product of the corresponding engrailed sequence. These data suggest that Mo-en.1 represents a mouse homolog of a gene of the Drosophila engrailed gene complex. Mo-en.1 has been mapped to chromosome 1, indicating it is not linked to other homeo box sequences that have been mapped in the mouse genome. Analysis of poly(A)+ RNA extracted from teratocarcinoma cells and whole mouse embryos demonstrates that the conserved homeo box region of Mo-en.1 is expressed differentially during mouse embryogenesis.

Animals↗

PubMed1985

Molecular probes define different regions of the mouse t complex.

Four genomic clones obtained from microdissected fragments of the proximal portion of mouse chromosome 17 have been used to identify a series of t-haplotype-specific restriction fragments. Their specificity is defined by presence in eight complete t haplotypes and absence from 18 inbred strains of wild-type mice. Partial t haplotypes contain subsets of the t-specific fragments, and each can be classified according to the t-specific fragments it contains. This is the first molecular evidence that independent partial t haplotypes contain different lengths of t haplotype DNA. Recombination studies indicate that partial t haplotypes suppress recombination in proportion to the extent of t haplotype DNA they contain. Molecular analysis of partial t haplotypes shows that the t-specific fragments map to and thus define different regions of the t complex. Certain regions of t haplotype DNA defined by t-specific restriction fragments can be correlated with loci involved in the control of transmission ratio distortion.

Alleles↗

PubMed1985

The extracellular matrix in development and in disease.

Recent concepts suggest that there are specific interactions between cells and the components of the extracellular matrix. These are mediated via cell surface receptors for the glycoproteins, collagens, and proteoglycans of the matrix. Two types of reactions are postulated. Normal matrix components, that is, those that are indigenous to the tissue, induce functional competence in the cells. Trauma, inflammation, or related reactions induce the production of matrix components more typical of fibrotic tissues. These matrix components cause a shift in cellular activities as they engage cell receptors. Various mechanisms can prolong and augment the fibrotic response, leading to an irreversible loss of tissue structures and function. New methods are available for identifying the cells and species of matrix proteins active in disease states.

Animals↗

PubMed1985

The C1q subunit of the first component of complement binds to laminin: a mechanism for the deposition and retention of immune complexes in basement membrane.

The C1q subunit of complement component C1 is known to bind to immune complexes, which often are deposited in basement membrane. We investigated the possibility that this deposition is a result of binding to laminin, a large basement membrane glycoprotein. C1q showed saturable binding to immobilized laminin; this binding was increased at reduced ionic strength. Intact C1 did not bind laminin. A ternary complex was formed by laminin, C1q, and aggregated IgG. This complex formation was dependent on and proportional to the amount of C1q bound to the aggregated IgG. Binding of laminin to C1q occurred with a Kd of 2 nM and was stronger than the binding of C1q to fibronectin. Preliminary data, including electron micrographs of rotary-shadowed preparations, suggest that laminin binds to the collagen-like tail of C1q. Electron microscopy localized the site of interaction with C1q to a short arm of laminin. Since laminin is found only in basement membranes, the interaction between laminin and C1q could be involved in the deposition and retention of immune complexes in these structures.

Antigen-Antibody Complex↗