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Biomedical subjects

J M Martin

Publications and source records attributed to J M Martin.

At least 109 records · Page 6Linked to original sources

Analysis of parathyroid hormone in bovine parathyroid cysts.

Parathyroid cysts were found in 2-5% of beef steers and heifers while collecting parathyroids at a local abattoir. The cyst fluid contained parathyroid hormone (PTH), averaging 15 ng/ml of fluid. Reversed-phase high-performance liquid chromatographic (HPLC) analysis of the fluid using an acetonitrile-trifluoroacetic acid solvent system, followed by carboxyl-terminal specific PTH radioimmunoassay of the eluant fractions, revealed two major peaks of PTH immunoreactivity. The first eluted at approximately 25% acetonitrile, a position similar to the commercially available human carboxyl terminal fragment of PTH-(39-84). The second peak, at approximately 37% acetonitrile, corresponded to the elution position for bovine PTH-(1-84). The cyst fluid was tested for enzyme activity by incubation of 125I-labeled bovine PTH with cyst fluid for 30 minutes at 37 degrees C, which resulted in no fragmentation of the labeled peptide. Immunohistochemistry of the cells lining the parathyroid cyst indicated that they contained PTH.

Animals↗

Could bovine serum albumin be the initiating antigen ultimately responsible for the development of insulin dependent diabetes mellitus?

Western blotting of either a cloned rat beta-islet tumour cell extract or isolated BB rat islets with rat anti-bovine serum albumin antiserum revealed a cross-reacting protein (Mr = 69,000). A protein of similar molecular size was observed by fluorography in proteins immunoprecipitated from islet cells labelled with (35S)-methionine using anti-bovine serum albumin antiserum. In comparing the primary structure of the beta subunits of the proteins Ia, DQ and DR a region of homology with bovine serum albumin became evident. Analysis of the amino-acid homology in relation to the DR/DQ allotypes found in the human population gave a strong correlation between the combined DR and DQ homology score with bovine serum albumin and the incidence of insulin dependent diabetes mellitus.

Adenoma, Islet Cell↗

Continuous negative pressure and intermittent mandatory ventilation in the management of pulmonary interstitial emphysema: a preliminary study.

We reviewed the clinical courses of 12 prematurely born newborns who were placed in continuous negative pressure (CNP) in an Isolette negative pressure ventilator for refractory hypoxemia while receiving intermittent positive pressure mandatory ventilation. All patients had severe lung disease as documented by an increased oxygenation index and bilateral pulmonary interstitial emphysema on x-ray examination. Patients were separated into two groups--survivors and nonsurvivors, with six patients in each group. Initiation of CNP resulted in a significant initial improvement in oxygenation in both groups seen as a 52% decrease in the oxygenation index in survivors and a 57% decrease in the oxygenation index in nonsurvivors (P less than .05). The survivors characterized themselves by showing a further sustained improvement in the oxygenation index--31.4 +/- 9.1 to 6.9 +/- 5.0 (P less than .01)--and a significant decrease in the mean airway pressure--11.6 +/- 4.6 cm H2O to 5.0 +/- 1.9 cm H2O (P less than .05). Four of the six survivors showed radiographic resolution of pulmonary interstitial emphysema. CNP was initiated at a mean age of 68.3 hours in the survivors. Nonsurvivors were initiated in CNP at a mean age of 134.3 hours, but went on to clinically deteriorate owing to irreversible hypoxemia and acidosis. Both oxygenation index and mean airway pressures were virtually unchanged compared with their initial values. The exact mechanisms by which CNP improves pulmonary function in this group of infants is unknown.(ABSTRACT TRUNCATED AT 250 WORDS)

Humans↗

Occupational risks: a follow-up method based on individual work history records including the environmental aspects of the workplace.

This article analyzes the Curriculum Laboris or career reconstruction as a means of studying the occupational risks of employees. A type of Curriculum Laboris which makes the established record easier to use has been suggested. The authors point out some advantages to be expected from the proposed computerized method. The usefulness of the proposed Curriculum Laboris is largely conditioned by lack of standards for guiding data collection by physicians. Finally, the Curriculum Laboris has been considered as an element of a larger information system.

Career Mobility↗

In-vivo effect of prostaglandin F-2 alpha treatment on secretory granules in the corpus luteum of the late pregnant cow.

Electron microscopy evaluation indicated that the large luteal cells from control (saline-injected) cows were morphologically similar: 42-72% of the large luteal cells, in which the nucleus was present in the tissue section, contained secretory granules (100-300 nm diameter). From cell to cell, clustering of granules was observed in various regions of the cell cytoplasm, ranging from locations next to the nucleus to those next to the cell membrane where exocytosis was noted. The large mitochondria containing dense inclusions (500-1800 nm diameter) were uniformly distributed throughout the cytoplasm of 98% of the large luteal cells. The morphology of large luteal cells at 2.5 and 5 min after PGF-2 alpha treatment (25 mg) was similar to that of saline controls. However, at 15 and 30 min after PGF-2 alpha the secretory granules were observed primarily at the cell periphery and at 15 min only 22% of the large luteal cells contained secretory granules. Exocytosis involving fusion of the granule and cell membrane was more evident at this time. At 60 min after PGF-2 alpha few large luteal cells (0.5%) were observed with small granules. We suggest that the secretory granules represent those organelles responsible for packaging secretory proteins for transport out of the cell, and PGF-2 alpha is an effector that will initiate cell depletion of these granules and their contents.

Animals↗

Solubilization of the active vasoactive intestinal peptide receptor from human colonic adenocarcinoma cells.

The non-ionic detergent n-octyl-beta-D-glucopyranoside was used to solubilize the VIP (vasoactive intestinal peptide) receptor from human colonic adenocarcinoma cell line HT29-D4. The binding of monoiodinated 125I-VIP to the solubilized receptor was specific, time-dependent, and reversible. Scatchard analysis of data obtained from competitive displacement of monoiodinated 125I-VIP by native VIP suggested the presence of two classes of VIP binding sites with Kd values of 0.32 and 46.7 nM. The binding capacities of these two classes were 1.7 x 10(10) and 30.2 x 10(10) sites/mg of proteins, respectively. The solubilized receptor retained the specificity of the human VIP receptor towards the peptides of the VIP/secretin/glucagon family. The order of potency in inhibiting monoiodinated 125I-VIP binding was VIP (IC50 = 1.0 x 10(-9) M) much greater than peptide histidine methionine amide (IC50 = 10(-7) M) greater than growth hormone-releasing factor (IC50 = 3 x 10(-7) M) greater than secretin (IC50 greater than 10(-6) M); glucagon had no effect on VIP binding. The reducing agent dithiothreitol inhibited in a dose-dependent manner the binding of 125I-VIP. Covalent cross-linking experiments between the solubilized receptor and 125I-VIP showed that after sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography two major and one minor polypeptides of Mr 67,000, 72,000, and 83,000 were specifically labeled. When analyzed by gel filtration, the n-octyl-beta-D-glucopyranoside-solubilized 125I-VIP-receptor complex was resolved into two major peaks with molecular mass in the range of 60-70 and 270-300 kDa. Thus, the soluble form of the VIP receptor was probably a multimeric complex in which disulfide bonds may play an important role to hold the receptor in an active configuration.

Adenocarcinoma↗

Modification of neuronal muscarinic receptor-mediated responses by islet-activating protein.

Islet-activating protein (IAP) was used to investigate the role of the guanosine triphosphate binding proteins Gi and/or Go in muscarinic acetylcholine receptor-mediated responses in neuroblastoma cells (clone N1E-115). Incubation of intact cells for 24 h with 20 ng/ml IAP resulted in inhibition of subsequent IAP catalyzed incorporation of [32P]ADP-ribose into a membrane protein doublet of molecular weight 40,000 (Gi alpha and Go alpha). IAP treatment fully blocked muscarinic receptor-mediated inhibition of cAMP accumulation. Incubation of intact cells with carbachol for 8 h resulted in the concentration dependent loss of membrane muscarinic receptor. Pretreatment of cells with IAP prior to carbachol exposure partially blocked the subsequent decrease in receptor number. Pretreatment of cells with IAP had no effect on the ability of carbachol to stimulate phosphoinositide hydrolysis in neuroblastoma cells. Thus, while the guanosine triphosphate binding proteins Gi and/or Go are involved in coupling the muscarinic receptor to some of the physiological responses in these cells, it is clear that activation of phospholipase C by the muscarinic receptor is a Gi/Go independent response.

Adenosine Diphosphate Ribose↗

Photoaffinity labelling of the vasoactive-intestinal-peptide-binding site on intact human colonic adenocarcinoma cell line HT29-D4. Synthesis and use of photosensitive vasoactive-intestinal-peptide derivatives.

N-Hydroxysuccinimidyl 4-azidobenzoate, a u.v.-sensitive heterobifunctional reagent, was used to synthesize photoreactive derivatives of the vasoactive intestinal peptide (VIP). Products of the reaction were purified by reverse-phase h.p.l.c. Three 4-azidobenzoyl-VIP (4-AB-VIP) derivatives were able to compete with monoiodinated 125I-VIP with an apparent KD of 2.5, 6.3 and 12.5 nM compared with 0.6 nM for native VIP. H.p.l.c.-purified mono[125I]iodinated VIP was used to synthesize 4-AB-125I-VIP derivatives. They were used to photoaffinity-label the VIP-binding site of HT29-D4 cells, a clone derived from the human colonic adenocarcinoma cell line HT29. Only one polypeptide, of Mr 70,000 +/- 5000 (mean +/- S.D.) was specifically labelled. The Mr of the component thus characterized was slightly higher than that of the major species (Mr 67,000) labelled after cross-linking experiments using 125I-VIP, conventional homobifunctional reagents and HT29 cells. Nevertheless, the specificity and extent of glycosylation of these two components were identical. These new photosensitive VIP derivatives should be useful tools with which to investigate further VIP-receptor structure and metabolism.

Affinity Labels↗

The vasoactive intestinal peptide (VIP) receptor: recent data and hypothesis.

Vasoactive intestinal peptide (VIP) is a neuropeptide with a broad range of biological activities in various tissues. After interaction with its membrane receptor, VIP generally induces a very large increase in the intracellular cyclic AMP level. Receptors for VIP have been described in numerous tissues and cell lines. The first results on VIP receptor structure have been obtained by covalent cross-linking using bifunctional reagents. The molecular mass of the different components characterized in this way differs greatly according to the species and the tissue used. This heterogeneity may reflect either a difference in the length of the cross-linked polypeptide backbone or differently glycosylated forms of the same polypeptide. The VIP binding site of intact human adenocarcinoma cells (HT29 cells) is an Mr 64,000 glycoprotein with 20kDa of N-linked oligosaccharide side chains containing sialic acid. The structure of the VIP binding site from HT29 cell is compared, first to the structure of the VIP receptor from other tissues, particularly that from rat liver, and second to the structure of the hepatic glucagon binding site. Recently, solubilization of the VIP receptor in an active form has provided a new way of studying this receptor. The HT29 cell line is an appropriate model to study the dynamics of the VIP receptor. After binding to its receptor, VIP is rapidly internalized, probably by receptor-mediated endocytosis. This internalization leads to a decrease in the cell surface receptor number and simultaneously to a homologous desensitization of adenylate cyclase. VIP is then degraded in the lysosomes, while most of the receptors are recycled back to the cell surface. The presence of an intracellular pool of unoccupied VIP receptors has been demonstrated after inactivation of the cell surface receptors by chymotrypsin. The kinetics of the receptor reappearance at the cell surface, after inactivation by chymotrypsin or after receptor-mediated endocytosis, indicate 2 possible intracellular pathways for occupied and unoccupied VIP receptors.

Acetylglucosaminidase↗

HT 29, a model cell line: stimulation by the vasoactive intestinal peptide (VIP); VIP receptor structure and metabolism.

HT 29, a cell line derived from a human colonic adenocarcinoma, is highly responsive to the vasoactive intestinal peptide (VIP) as shown by a more than 100-fold intracellular cAMP increase (Ka = 0.3 nM), the stimulations of protein kinase A (Ka = 0.1 nM) and the low-Km cAMP phosphodiesterase (Ka = 40 nM). Remarkably, adenylate cyclase, cAMP-dependent kinase and cAMP-specific phosphodiesterase are activated in a sequential manner. Binding studies with [125I]-labeled VIP indicate a high affinity site with a Kd value (0.5 nM) close to the activation constant value (Ka) of the three enzymes. The molecular structure of the VIP receptor was studied by immunological and chemical approaches. A monoclonal antibody (mAb 109-10-16) which partially decreased the binding of VIP to its receptor allowed the characterization of Mr = 53,000 and Mr = 48-49,000 polypeptides. More precise identification of protein components of the VIP receptor resulted from covalent cross-linking on intact HT 29 cells by four bifunctional reagents: dithiobis-(succinimidyl propionate) and its non-cleavable analog disuccinimidyl suberate, the photoactivable azido phenyl glyoxal and dimethylpimelimidate. Analysis by SDS-polyacrylamide gel electrophoresis demonstrated a major band of Mr = 67,000 regardless of which cross-linker was used. The same band and an Mr = 49,000 species were found in experiments using a crude membrane fraction of HT 29 cells. Assuming one molecule of VIP (Mr = 3326) linked per polypeptide, these observations suggest that an Mr = 64,000 species belongs to the VIP specific plasma membrane receptor. This protein contains an Mr = 20,000 N-linked sialic acid rich oligosaccharidic moiety.(ABSTRACT TRUNCATED AT 250 WORDS)

3',5'-Cyclic-AMP Phosphodiesterases↗

Multicore myopathy: not always a benign entity.

Four patients with Multicore Myopathy, a rare morphologically distinct myopathy, are described. Although previously considered to be a non-progressive or only slowly progressive myopathy, progression to significant disability was seen in three of our cases. The association of cardiac disease with Multicore Myopathy has not been previously emphasised. All four patients in this study had a cardiomyopathy, and heart disease was the cause of death in two of the patients. Multicore Myopathy is not always a benign entity. Cardiac involvement, when present, adversely affects prognosis.

Adolescent↗

Hairy cell leukemia and anti-leukocyte common antigen.

Thirty-three bone marrow biopsies from 15 patients with hairy cell leukemia (HCL) were evaluated morphologically and immunohistochemically by use of the peroxidase-antiperoxidase technique to demonstrate reactivity for leukocyte common antigen (LCA). Hairy cells in all biopsies showed a distinctive and characteristic pattern for LCA, which decorated the periphery of the cytoplasm but that left most of the cytoplasm and the nucleus unstained. Anti-LCA was particularly helpful in highlighting focal or subtle leukemic infiltrates. Hairy cells in biopsies from three patients had, on routine morphologic examination, a spindled and sarcomatoid appearance, but these too were strongly LCA positive. Treatment regimens were variable: five patients had splenectomy and received chemotherapy; five patients had splenectomy alone; and four patients had chemotherapy alone. Seven patients received interferon, and one patient received no treatment. In those six patients who had multiple biopsies as part of follow-up examinations, hairy cells as identified by anti-LCA were continuously present. Often in significant numbers, and were usually underestimated or not identified by routine examination. In those patients who received chemotherapy, qualitative alterations in the LCA reaction of hairy cells were observed.

Adult↗

Appearance of prostaglandins, thromboxane B2, and hepoxilin A3 in the circulation of the normal and diabetic (BB) rat after arachidonic acid administration--correlation with plasma insulin.

The temporal in vivo expression of the eicosanoids (products of the cyclooxygenase pathway and one product of the 12-lipoxygenase pathway, hepoxilin A3) was investigated after bolus intravenous injection of arachidonic acid in the normal rat and in the genetic rat model of type I insulin-dependent diabetes, the diabetic BB rat. The temporal relationship between the expression of these products and plasma insulin concentrations was also investigated to determine whether any correlation existed between the rise in plasma insulin levels and any of the newly formed eicosanoids. Measurements of the eicosanoids present in whole blood were carried out using the deuterium isotope dilution technique involving separation of pentafluorobenzyl esters, O-methyl oximes, and trimethylsilyl ether derivatives by high-resolution gas chromatography and specific detection by negative ion chemical ionisation mass spectrometry in the selected ion mode. Injection of arachidonic acid resulted in large and statistically significant increases in the blood concentrations of all products within 1 min, with thromboxane B2 (the stable product of thromboxane A2) and trioxilin A3 (the stable product of hepoxilin A3) being the highest (4.5-12 ng/mL). The mean concentrations of thromboxane B2 and trioxilin A3 in blood appeared greater in the diabetic BB rat than in the normal rat, while the opposite was found for 6-keto PGF1 alpha (the stable product of prostacyclin). The apparent greater ratio of thromboxane B2 to 6-keto PGF1 alpha in the diabetic BB rat than in the normal rat supports a prothrombotic nature of platelets associated with diabetes.

6-Ketoprostaglandin F1 alpha↗

Time-dependent change in electrophysiologic milieu after myocardial infarction in conscious dogs.

This study was designed to assess the time-dependent change in propensity to induction of malignant ventricular tachyarrhythmia after myocardial infarction. Instrumented conscious dogs were assessed during serial drug-free electrophysiologic studies over 26 +/- 9 days (range 17 to 35 days) after 2 hr occlusion-reperfusion of the left anterior descending coronary artery. Of the 19 animals studied, 11 continued to have sustained ventricular tachyarrhythmias inducible (group I) over this time period. In the eight remaining animals, spontaneous loss in the ability to induce sustained ventricular tachycardia occurred (group II). Myocardial infarct size in group I animals (18 +/- 8%) was significantly greater than that in group II dogs (12.5 +/- 5%; p less than .05). Even in group I animals, time-dependent changes occurred in the number of extrastimuli required to induce ventricular tachycardia and the frequency with which left ventricular stimulation was necessary. A differential pattern of time-dependent changes in electrophysiologic variables was observed when comparing group I and II animals. The conduction time to the infarct zone was prolonged during follow-up in group I animals, while in group II animals this variable was unchanged. Repolarization time recorded in the border zone remained unchanged in group I animals, but it was significantly shortened in group II animals. In addition, ventricular effective refractory period in the infarct zone shortened over time in group I animals but did not change in group II animals. In conclusion, time-dependent changes occur in electrophysiologic variables that are associated with a progressive decrease in propensity to induction of ventricular tachycardia after myocardial infarction. A critical determinant of whether propensity to ventricular tachycardia resolves over time is size of myocardial infarction.

Animals↗