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Biomedical subjects

J L Martin

Publications and source records attributed to J L Martin.

At least 271 records · Page 15Linked to original sources
PubMed1989

Structure of the Mr 140,000 growth hormone-dependent insulin-like growth factor binding protein complex: determination by reconstitution and affinity-labeling.

To determine the structure of the high molecular weight, growth hormone-dependent complex between the insulin-like growth factors (IGF-I and IGF-II) and their binding proteins in human serum, we have reconstituted the complex from its purified component proteins and analyzed it by gel electrophoresis and autoradiography after covalent cross-linking. The proteins tested in reconstitution mixtures were an acid-labile Mr 84,000-86,000 glycoprotein doublet (alpha subunit), an acid-stable Mr 47,000-53,000 glycoprotein doublet with IGF-binding activity (BP-53 or beta subunit), and IGF-I or IGF-II (gamma subunit). In incubations containing any one of the three subunits 125I-labeled and the other two unlabeled, identical 125I-labeled alpha-beta-gamma complexes of Mr 140,000 were formed. Minor bands of Mr 120,000 and 90,000 were also seen, thought to represent a partially deglycosylated form of the alpha-beta-gamma complex, and an alpha-gamma complex arising as a cross-linking artifact. When serum samples from subjects of various growth hormone status were affinity-labeled with IGF-II tracer, a growth hormone-dependent Mr 140,000 band was seen, corresponding to the reconstituted alpha-beta-gamma complex. Other growth hormone-dependent labeled bands, of Mr 90,000 (corresponding to alpha-gamma), Mr 55,000-60,000 (corresponding to labeled beta-subunit doublet), and smaller bands of Mr 38,000, 28,000, and 23,000-25,000 (corresponding to labeled beta-subunit degradation products), were also seen in the affinity-labeled serum samples and in the complex reconstituted from pure proteins. All were immunoprecipitable with an anti-BP-53 antiserum. We conclude that the growth hormone-dependent Mr 140,000 IGF-binding protein complex in human serum has three components: the alpha (acid-labile) subunit, the beta (binding) subunit, and the gamma (growth factor) subunit.

Acromegaly↗
PubMed1989

Sexual behavior changes and HIV antibody in a cohort of New York City gay men.

A sample of 357 gay/bisexual men, ages 20 to 65, were interviewed in 1985 regarding their sexual patterns over the course of the AIDS (acquired immunodeficiency syndrome) epidemic. Within four months after the interview, men were tested for antibody to HIV (human immunodeficiency virus) and 36 per cent were found positive. Cessation of receptive anal intercourse was associated with HIV negative antibody status, (OR greater than 3.0). Reducing the number of sexual partners, becoming monogamous, or eliminating other forms of sexual contact were not related to risk of HIV.

Acquired Immunodeficiency Syndrome↗
PubMed1989

Release of vasoactive intestinal peptide in mouse cerebral cortex: evidence for a role of arachidonic acid metabolites.

In rodent cerebral cortex, vasoactive intestinal peptide (VIP) is contained in a homogeneous population of radially oriented bipolar interneurons. We have observed that 4-aminopyridine (4-AP), a K+-channel blocker, promotes a concentration- and Ca2+-dependent release of VIP from mouse cerebral cortical slices, with a significant effect already observed at 50 microM. Over 70% of VIP release elicited by 4-AP is blocked by 2 microM tetrodotoxin (TTX). Mepacrine, an inhibitor of phospholipase A2 (PLA2) activity and hence of arachidonic acid (AA) formation from membrane phospholipids, markedly inhibits (IC50 of approximately 15 microM) the release of VIP evoked by 4-AP. The inhibitory effect of mepacrine is not additive to that of TTX, thus indicating an involvement of PLA2 activation in the TTX-sensitive component of the 4-AP-evoked release. As a corollary, melittin (0.1-10 micrograms/ml), a PLA2 activator, promotes VIP release. Inhibition of AA metabolites of the lipoxygenase pathway by nordihydroguaiaretic acid, 5,8,11,14-eicosatetranoic acid, and caffeic acid results in a concentration-dependent inhibition of VIP release evoked by 4-AP. This set of observations indicates for the first time that the formation of AA metabolites of the lipoxygenase pathway plays a role in the release of a peptide in the mammalian CNS. Furthermore, these observations together with the previously reported potentiation by prostaglandins of the increase in cyclic AMP elicited by VIP in mouse cerebral cortex (Schaad et al., 1987) indicate that AA metabolites may act at both the presynaptic (lipoxygenase metabolites) and the postsynaptic (cyclooxygenase metabolites) levels to increase the "throughput" or "strength" of VIP-containing circuits in the rodent neocortex.

4-Aminopyridine↗
PubMed1989

[Bioluminescence assay of luteinizing hormone in plasma and urine].

The authors describe a bioluminescent immunoassay of LH in plasma and urine. It uses two monoclonal antibodies, one is labelled with glucose-6-phosphate-dehydrogenase, the other one is coimmobilized together with bioluminescent enzymes from marine bacteria on the same adsorbent (Sepharose). This assay can be performed directly on 20 microliters plasma or 10 microliters urine. The protocol is very fast, no separation step is required to remove the excess labeled antibodies. The inhibitory effect of the biological sample on luminescent reaction is determined by adding NADH to the assay tubes. The working range of this assay is 3 to 300 Ul/l, with a sensitivity of detection of 0.5 Ul/l. Recovery, linearity, within and between assay precision were evaluated and appeared to be satisfactory. The authors have observed a good correlation between results obtained with our method and with radioimmunoassay.

Female↗
PubMed1988

[Reversal of positive serology for human immunodeficiency virus (HIV). Apropos of 2 case reports].

The medical history and laboratory data of two persons who have been in contact with HIV-I positive carrier individuals are reported. The serologic data of successive serum samples collected from these persons were first negative, then positive (including antibodies directed against Gag and Env gene products), and finally negative for presence of anti-HIV-I antibodies. Physical examination and other laboratory data, including absolute number of CD4 lymphocytes/mm3, were within the normal range in these two individuals, who presented a reversal of positive serology for HIV-I.

AIDS Serodiagnosis↗
PubMed1988

Channels at the catalytic site of glycogen phosphorylase b: binding and kinetic studies with the beta-glycosidase inhibitor D-gluconohydroximo-1,5-lactone N-phenylurethane.

Regions of low packing density in the vicinity of the catalytic site of glycogen phosphorylase b are described with the aid of a computer program that generates a contour map in which the contour level is inversely proportional to the packing density in the protein. It is shown that, although there is no direct route from the catalytic site to the surface, there are two possible channels that could allow access for substrates following conformational changes in the enzyme. The first channel, channel 1, leads from the catalytic site to the surface close to the nucleoside inhibitor site and requires movements of residues 280-285 and Arg 569 in order to obtain access. Previous crystallographic experiments have shown that in the presence of substrates or R-state inhibitors these parts of the polypeptide chain undergo large conformational changes. The properties of the second channel (channel 2), which is the more extensive channel, have been investigated with the potent beta-glycosidase inhibitor D-gluconohydroximo-1,5-lactone N-phenylurethane (PUG). Crystallographic binding studies at 2.4-A resolution show that the compound binds neatly at the catalytic site of phosphorylase b. The glucopyranosylidene ring, in the half-chair conformation, occupies a similar but not identical position (shift about 0.6 A) to that occupied by other glucosyl compounds bound at the catalytic site.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1988

Risk-benefit ratio of sodium fluoride treatment in primary vertebral osteoporosis.

The risk-benefit ratio of combined fluoride-calcium therapy in primary vertebral osteoporosis was examined prospectively in patients with at least one vertebral fracture. 257 patients were randomised to receive sodium fluoride 25 mg twice daily plus elemental calcium 1 g daily and a vitamin D2 supplement, and 209 received one of the alternative therapies usually prescribed in France. After a follow-up of 24 months the fluoride-calcium group showed a significantly lower rate of new vertebral fractures, the main adverse effect of the regimen being a higher incidence of osteoarticular pains in the ankle and foot; the risk of non-vertebral fractures was not increased, and digestive disorders arose with equal frequency in the two groups. Sodium fluoride 50 mg daily seems to represent a reasonable compromise in terms of anti-fracture effectiveness and side-effects.

Administration, Oral↗
PubMed1988

Photophysics and reactivity of heme proteins: a femtosecond absorption study of hemoglobin, myoglobin, and protoheme.

On the basis of our time-resolved absorption measurements of hemoglobin (Hb), myoglobin (Mb), and protoheme (PTH), either unligated or ligated with CO, O2, or NO, we propose a description of the photophysics of heme proteins that encompasses their photodissociation, the origin and fate of the observed short-lived transients, and the appearance of the ground-state, unligated heme proteins. Two distinct species are formed upon ligand photodissociation, which occurs in less than 50 fs. We assign these species to excited states of the unligated heme and label them (for the case of hemoglobin) as Hb*I and Hb*II. We suggest that Hb*I is already at least partially domed and has a spin state of at least S = 1. Hb*I decays in 300 fs to the ground-state unligated heme species, which we consider to be S = 2 and at least partially domed. The population of Hb*II varies with the ligand. It is more significant when the ligand is O2 or NO than when the ligand is CO. The similarities of the picosecond and femtosecond bleaching and absorption kinetics of HbCO with those of PTHCO (and of HbNO with those of PTHNO) indicate that in this time domain the importance of steric features of the protein are less important than the nature of the ligand itself in the geminate recombination process as well as in the relative amounts of the two heme excited states created. It is suggested that the quantum yield of ligand photodissociation is unity whether the ligand is O2, NO, or CO. The low yield of photodissociated heme-O2 or heme-NO compounds as measured on the microsecond time scale is thus attributed to a fast (2.5 ps) recombination of O2 or NO with Hb*II. We discuss geminate recombination measurements of cyanomet hybrid hemoglobins with NO and consider these results in terms of alpha and beta subunit heterogeneity. The first picosecond transient absorption spectra of cyanomet-CO hybrid hemoglobins are presented and are compared with the spectra of other heme compounds. The superimposability of the transient spectra on the equilibrium spectra of heme compounds that exhibit minimal or no cooperativity is noted and is compared with the case of cooperative systems where the transient spectra are distorted with respect to the equilibrium spectra. This distortion is interpreted in terms of an interaction of a domed heme with the F helix.

Heme↗
PubMed1988

Stimulation by nicotine of enteric inhibitory nerves and release of vasoactive intestinal peptide in the taenia of the guinea-pig caecum.

The stimulation by nicotine of intramural nerves and the role of ATP and vasoactive intestinal peptide (VIP) as inhibitory transmitters were studied in the isolated taenia of the guinea-pig caecum. Nicotine (4-32 microM) caused transient, concentration-dependent relaxations which were unaffected by atropine, prazosin or sotalol. Drugs with membrane-stabilizing activity, such as dl-propranolol (0.5 microM), d-propranolol (0.5 microM) or lidocaine (10 microM) antagonized the nicotine-induced relaxation without modifying the response to electrical field stimulation. Similar results were obtained with the cyclooxygenase inhibitor, indomethacin (2.8 microM). Nucleotide pyrophosphatase (0.5 U/ml), which hydrolyzes ATP to AMP, reversibly inhibited the response to nicotine but the response to field stimulation was not decreased. Nicotine evoked a calcium-dependent release of VIP, which was blocked by tetrodotoxin (1 microM), d-propranolol (0.5 microM) or, as previously shown, by apamin (0.2 microM). The finding that nicotine-induced relaxation was accompanied by the neuronal release of VIP is compatible with the possibility that VIP is an inhibitory transmitter but is not definitive evidence, since it could have been due to the stimulation of distinct populations of nerves by nicotine.

Animals↗
PubMed1988

Queensland fruit fly virus, a probable member of the Picornaviridae.

A picornavirus was isolated from various life stages of the Queensland fruit fly, Dacus tryoni. This virus, Queensland fruit fly virus (QFFV) has virions with a diameter of 30 nm and a sedimentation coefficient of 178 S. One third of the particles in preparations were empty capsids or natural top component (NTC) with a sedimentation coefficient of 95 S. The buoyant density (rho) of virions and NTC in CsCl was 1.34 and 1.30 g/ml respectively; small amounts of a dense component (rho = 1.45 g/ml) were also detected. The capsid contained three major protein species of molecular weight (mol.wt.) 41,700, 36,500, and 31,300, in approximately equimolar proportions. NTC contained three major species of mol. wt. 44,700, 41,700, and 31,300. The nucleic acid present only in the bottom component virions was RNA and comprised about 30% of the particle weight and had a mol. wt of 2.88 kd, contained a poly(A) tract, and had a base ratio: G = 20; A = 32; C = 15; U = 33. The mol. wt. of the virion was estimated to be approximately equal to 9.5 kd. When virions were heated at 56 degrees C and above, they converted into artificial top component (ATC), which had the same protein composition as the virion when analysed by SDS-PAGE. In immunodiffusion tests the virions and NTC were indistinguishable, but a minor difference in antigenicity was detected between the virions and ATC. Virions were stable between pH 3 and 9 inclusive, and between 5 and 7 in the presence of 0.14 M NaCl. Immunodiffusion tests showed that QFFV was serologically unrelated to a range of picornaviruses as well as an unclassified virus isolated from the Mediterranean fruit fly, Ceratitis capitata. The data show that QFFV is probably a member of the Picornaviridae, genus Enterovirus.

Animals↗
PubMed1988

High- and low-affinity binding sites for vasoactive intestinal peptide (VIP) in the rat kidney revealed by light microscopic autoradiography.

The distribution of VIP binding sites in rat kidney and adrenal gland has been examined by light microscopic autoradiography. A fully characterized mono-iodinated molecular form of VIP (M-125-I-VIP) which maintains the biological activity of the native peptide, was used for this study. Two types of VIP binding sites, with high and low affinity, have been identified. High affinity sites are associated with (i) glomerular structures in the cortex, (ii) the inner stripe of the outer medulla, possibly corresponding to Henle's loops and distal tubules, (iii) radiated structures in the inner zone of the medulla, likely to represent labeling of collecting ducts and/or vascular bundles and (iv) the adrenal cortex. Autoradiographic grains associated with low affinity sites are present diffusely throughout the renal cortex, possibly corresponding to labeling of tubular and/or vascular structures, and throughout the adrenal gland. These observations further delineate a role of VIP in renal and neuroendocrine function.

Animals↗
PubMed1988

Insulin-like growth factor-binding proteins (IGF-BPs) produced by human skin fibroblasts: immunological relationship to other human IGF-BPs.

We have characterized the insulin-like growth factor-binding protein (IGF-BP) produced by neonatal human skin fibroblasts in monolayer culture using antibodies specific for the acid-stable subunit of the 150K GH-dependent IGF-BP complex, BP-53, and the amniotic fluid IGF-BP, BP-28. Fibroblasts produced 65.3 +/- 10.4 ng/ml.72 h (SE; n = 6) immunoreactive BP-53 in serum-free medium; this was stimulated by increasing fetal bovine serum in the medium up to 385.3 +/- 49.0 ng/ml.72 h at 10% serum. Epidermal growth factor (EGF) also caused dose-dependent stimulation of BP-53 production, with a maximal effect (3-fold increase) at 30 ng/ml EGF. No immunoreactive BP-28 production was detectable in the presence or absence of serum or EGF. Neutral gel chromatography of serum-free medium revealed a peak of immunoreactive BP-53 at about 50K, with a smaller species at 20-30 K. Serum- and EGF-stimulated cells produced higher levels of about 50K BP-53, and an additional peak of immunoreactivity at 150K was present in serum-stimulated, but not EGF-stimulated, samples. Comparison of IGF-I and IGF-II binding by fibroblast BP-53 revealed slightly higher IGF-II than IGF-I binding, and association constants of 3-4 x 10(10) liter/mol for both IGFs, similar to BP-53 from human plasma. Affinity labeling of acid-stripped medium followed by nonreduced sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed specifically cross-linked IGF-binding species of 60K (identical to labeled plasma BP-53), 42K, and 37K. Only the 60K and 42K complexes were precipitable by antiserum to plasma BP-53, and none was precipitable by anti-BP-28 serum, suggesting that the 37K band might represent a third class of IGF-BP. We conclude that neonatal skin fibroblasts produce no BP-28, but do produce two IGF-BPs immunologically homologous to human plasma BP-53, one of which shows size and IGF-binding characteristics identical to the plasma protein.

Antibodies↗
PubMed1988

Cloning and expression of the growth hormone-dependent insulin-like growth factor-binding protein.

N-terminal as well as internal amino acid sequence data were obtained from the GH dependent, insulin-like growth factor (IGF) binding protein, BP-53, purified from human plasma. Based on these sequence data, full-length cDNA clones of BP-53 have been isolated, and the complete deduced sequence of BP-53 determined. This sequence contains a 27 amino acid putative signal sequence followed by a mature protein of 264 amino acids containing 18 cysteine residues clustered near the N- and C-terminus. The deduced protein sequence of BP-53 has 33% amino acid identity including conservation of all 18 cysteine residues with the recently cloned BP-28, a smaller human IGF-binding protein identified in amniotic fluid and also secreted by the cell line HEP G2. Expression of the cloned BP-53 cDNA in mammalian tissue culture cells results in secretion of the protein into the culture medium. This expressed protein is identical to plasma-derived BP-53 in its immunoreactivity, high affinity binding of IGF-I and IGF-II, and mobility on sodium dodecyl sulfate gel electrophoresis.

Amino Acid Sequence↗
PubMed1988

Predicting fertility of dairy bulls by inducing acrosome reactions in sperm with chondroitin sulfates.

Bull spermatozoa were incubated with 0 or 50 micrograms/ml chondroitin sulfates to induce acrosome reactions in vitro. Data were computed as percent increase in acrosome reactions elicited by chondroitin sulfates compared with control samples for each replicate. Data from 8 bulls with known nonreturn rates served as basis to predict nonreturn rates of 18 bulls entering an AI lineup. The average assay CV was 16.1% for all 26 bulls. Correlation coefficients of polynomial regression lines computed between actual nonreturn rates and percent increases in acrosome reactions in response to chondroitin sulfates were .98 and .81 for the 8 reference bulls and combined data of all 26 bulls, respectively. It is concluded that a laboratory assay using chondroitin sulfates to induce acrosome reactions in sperm can be used to predict relative fertility of bulls.

Acrosome↗