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M Costa

Publications and source records attributed to M Costa.

At least 469 records · Page 26Linked to original sources
PubMed1984

Analysis of the induction of alkali sensitive sites in the DNA by chromate and other agents that induce single strand breaks.

CaCrO4 was shown to induce alkali labile sites in the DNA of Chinese hamster ovary cells by analysis of the lack of linearity in the alkaline elution curves, and by a study of the dependence upon pH in the elution of DNA from filters. The effect of CaCrO4 on these parameters was compared with N-methyl-N-nitrosourea, an agent known to produce alkali labile sites. Based upon the aforementioned parameters HgCl2, formaldehyde and X-rays caused the formation of frank single strand breaks with little or no induction of alkaline labile sites. These findings demonstrate differences in the production of alkali sensitive sites by agents that cause DNA single strand breaks.

Alkalies↗
PubMed1984

Electrophysiology and enkephalin immunoreactivity of identified myenteric plexus neurones of guinea-pig small intestine.

Intracellular injection of the fluorescent dye, Lucifer Yellow CH, revealed the shapes of neurones in the myenteric plexus of the guinea-pig ileum, and these shapes were correlated with the electrophysiological properties and enkephalin immunoreactivity of the neurones. A total of eighty-three neurones were filled using electrodes containing a 5% solution of the dye. Forty-six cells had many short processes and a single long process (Dogiel type 1) and twenty-four cells had essentially smooth somas and one to eight long processes (Dogiel type II). Thirteen cells could not be put into either group. Enkephalin-like immunoreactivity was detected in twenty-two of the forty-six Dogiel type I cells. Eighteen of these had club-like short processes. No other cells of the eighty-three showed enkephalin-like immunoreactivity. Electrodes filled with a 0.5% solution of Lucifer Yellow in 0.5 M-KCl were used to record from and simultaneously to inject dye into 240 neurones. Eighty-six nerve cells had a slow after-hyperpolarization following the action potential (AH cells) and forty-six nerve cells had no after-hyperpolarization but exhibited a fast excitatory synaptic potential (S cells). The other cells could not be unequivocally identified by their observed electrophysiological characteristics. Almost all S cells (forty-two of forty-six) were Dogiel type I, while eighty-two of the eighty-six AH cells were Dogiel type II. Fifty S cells (eight located geometrically, forty-two by dye injection) and ninety-one AH cells (twenty-six located geometrically, sixty-five by dye injection) were examined for enkephalin immunoreactivity. Fifteen of the S cells were reactive, whereas all of the AH cells were unreactive. It appears that prolonged impalements reduce immunoreactivity so that the proportion of reactive neurones in this series is an underestimate of the true proportion of S cells with enkephalin-like immunoreactivity. The results suggest that a substantial proportion of the S cells in myenteric ganglia contain enkephalin immunoreactivity while none of the AH cells do. The enkephalin neurones have a distinctive shape and are all Dogiel type I cells. AH cells are nearly always Dogiel type II.

Action Potentials↗
PubMed1984

Neuropeptides contained in peripheral cardiovascular nerves.

The neuropeptides, substance P, vasoactive intestinal peptide (VIP), neuropeptide Y and enkephalin have been found in nerves associated with the heart and blood vessels of a range of mammals, including man. There is also evidence for some cardiovascular nerves with gastrin releasing peptide and neurotensin immunoreactivity. Substance P is in sensory nerves with a widespread distribution to the heart and all vascular beds. In general, large arteries have the densest innervation and the density of nerves decreases as arterial size decreases. In adult guinea-pigs, an adequate treatment with capsaicin causes the degeneration of almost all cardiovascular substance P nerves. Using capsaicin as a tool it has been shown that the substance P containing sensory nerves are not essential for baroreceptor reflexes. VIP nerves also have a widespread distribution, being particularly prominent in the cerebral arteries, uterine arteries and arteries of erectile and secretory tissues. Neuropeptide Y is located in the same cardiovascular nerves as noradrenaline. It is depleted from the nerves by reserpine or 6-hydroxydopamine. Enkephalin nerves have been reported with small arteries in only a few vascular beds.

Animals↗
PubMed1984

Mechanism of HgCl2 cytotoxicity in cultured mammalian cells.

Treatment of intact Chinese hamster ovary cells with HgCl2 produced a rapid, concentration-dependent induction of DNA single-strand breaks (SSB) as revealed by alkaline elution analysis. Direct addition of HgCl2 to cell lysates did not result in DNA strand breaks. HgCl2 treatment of cells also caused a rapid leakage of superoxide radicals that were detected in their media by measurement of the reduction of exogenously added cytochrome c. There was a linear relationship between the production of radicals and the induction of DNA strand breaks, and there were also excellent temporal correlations in these parameters. Addition of oxygen radical scavengers, such as the enzymes superoxide dismutase and catalase, to the extracellular media significantly reduced the extent of DNA damage caused by HgCl2 without a similar attenuation of its uptake into cells, as did the autoclaved enzymes. Similarly, addition of radical scavengers such as glycerol or ascorbate inhibited the DNA damage but also reduced the uptake of the metal by almost the same degree. Thus, because of secondary effects on uptake of the metal, the radical scavenger experiments could not address the importance of oxygen radicals in the DNA damage caused by HgCl2. SSB were enhanced when cells were treated with HgCl2 and diethylmaleate or diethyldithiocarbamate, agents that deplete cellular reduced glutathione or inhibit the intracellular activity of superoxide dismutase, respectively. Thus, DNA damage in cells rendered sensitive to radicals was greater when these cultures were subsequently treated with HgCl2. The binding of 203HgCl2 to the DNA of intact Chinese hamster ovary cells was also studied. These studies were made possible by the relatively high stability of Hg(II) interaction with DNA and by utilizing a gentle method of DNA isolation that minimized redistribution of intracellular Hg(II) complexes after cells were lysed. The amount of Hg(II) bound to DNA varied from approximately 7 to 35 Hg atoms per 10(4) base pairs (bp) at concentrations of HgCl2 that have been previously shown to produce between 1 SSB/10(7) bp and 1 SSB/10(6) bp. The Hg(II)-DNA adducts were relatively stable complexes, since they resisted treatment with 0.1 M EDTA and 1 M NaCl and were stable to precipitation of the DNA with ethanol and trichloroacetic acid. However, the Hg(II) was released from the DNA when it was degraded enzymatically to mononucleosides, suggesting that the Hg(II)-DNA bonds formed in the cell were not truly covalent and that the strength of Hg(II) binding to DNA depended upon polynucleotide structure.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1984

Substance P-containing nerves in the human small intestine. Distribution, ultrastructure, and characterization of the immunoreactive peptide.

Light and electron microscopic immunocytochemical techniques were used to examine the distribution and ultrastructure of substance P-immunoreactive nerves in human jejunum and distal ileum. The organization of human enteric substance P-containing nerves closely resembled that in other species. Dense arrays of varicose immunofluorescent fibers occurred in myenteric and submucous ganglia (which contained immunoreactive nerve cell bodies) and in the mucosa. There were fibers in both muscle layers, in the muscularis mucosae, and around blood vessels. Fibers in the myenteric plexus contributed to both ascending and descending pathways. Substance P-immunoreactive axon profiles contained small round and large round vesicles and were apposed to nerve cell bodies, and nonimmunoreactive and immunoreactive axon profiles. Synapselike contacts were occasionally observed on nerve cell bodies and processes. The substance P-like material was characterized by high pressure liquid chromatography and radioimmunoassay and found to be indistinguishable from the authentic undecapeptide. These results suggest that enteric nerves containing substance P may play similar roles in humans as in other species.

Adult↗
PubMed1984

Origins of peptide and norepinephrine nerves in the mucosa of the guinea pig small intestine.

Norepinephrine, acetylcholine, and certain peptides are contained in mucosal nerves and have potent effects on transepithelial water and electrolyte fluxes. It is difficult to ascribe roles for these nerves as their sources are unknown. The present studies were undertaken to determine the origins of nerve fibers that are found in the mucosa of the guinea pig small intestine and which contain one of the following substances: vasoactive intestinal peptide, substance P, somatostatin, neuropeptide Y, cholecystokinin, or norepinephrine. Nerve fiber origins were ascertained by making lesions to sever pathways through which the nerves could reach the mucosa. The lesioning operations were homotopic autotransplants of short (2 cm) segments of intestine; myectomies, in which a 5-10-mm length of intestine was stripped of longitudinal muscle and myenteric plexus; and extrinsic denervation, in which nerves reaching the intestine through the mesentery were severed. The results of these studies, considered along with previously published work, led to the upcoming conclusions. Nerve fibers in the mucosa showing immunoreactivity for vasoactive intestinal peptide, somatostatin, cholecystokinin, and neuropeptide Y arise from cell bodies in the overlying submucous plexus. Substance P fibers arise in part from the overlying submucous plexus and in part from the overlying myenteric plexus. Mucosal norepinephrine fibers arise from extrinsic sympathetic ganglia. Enkephalin, gastrin-releasing peptide, and 5-hydroxytryptamine, which are in some enteric nerves, are not found in submucous nerve cells and few, if any, fibers containing these substances supply the mucosa. Thus, the mucosa receives a dense nerve supply, much of which arises locally from submucous ganglia.

Adrenergic Fibers↗
PubMed1984

Effects of orally administered ritodrine on carbohydrate and lipid metabolism in pregnant patients with abnormal glucose tolerance.

The metabolic effects of a widely used betamimetic agent (ritodrine) were studied in two groups of pregnant patients with abnormal glucose tolerance. No significance modification of various metabolic parameters was detected when the drug was orally administered in standard doses for at least 1 week. The possible reasons for this finding are discussed. Following the absence of any diabetogenic effect, a prolonged use of betamimetics to achieve tocolysis seems to be safe even in patients with metabolic disorders.

Carbohydrate Metabolism↗
PubMed1984

Noradrenergic nerves in human small intestine. Distribution and ultrastructure.

Noradrenergic nerves in human jejunum and distal ileum were identified at the light microscopic level by fluorescence histochemistry and at the ultrastructural level by 5-hydroxydopamine loading in vitro and by the chromaffin reaction. The myenteric and submucous ganglia contained dense arrays of varicose noradrenergic fibers; the muscularis externa, muscularis mucosae, intestinal glands, and villi were sparsely supplied. Enteric axon profiles that had taken up 5-hydroxydopamine contained variable proportions of flattened, small round, and pleomorphic large vesicles all containing electron-dense material. Chromaffin-reactive enteric axon profiles and some enteric profiles in conventionally fixed untreated intestine contained similarly shaped vesicles. After 5-hydroxydopamine loading, noradrenergic nerves supplying blood vessels contained large round and small round vesicles with dense cores. Noradrenergic varicosities were concentrated near the edges of myenteric ganglia and were randomly distributed in submucous ganglia. Noradrenergic axons occasionally formed synapses on nerve processes in both myenteric and submucous ganglia, and on rare occasions, on nerve cell bodies in myenteric ganglia. Nonnoradrenergic axon profiles containing small round vesicles with granular inclusions (ring vesicles) were also found in human small intestine. The distribution and ultrastructure of noradrenergic nerves in human small intestine is similar to that in other mammals except for the occurrence of synapses in human ganglia.

Adrenergic Fibers↗
PubMed1983

Neurones localized with antibodies against choline acetyltransferase in the enteric nervous system.

This is the first report of the histochemical localization of peripheral neurones with antibodies raised against choline acetyltransferase. The antiserum was raised in a rat against the enzyme purified from porcine brain. An indirect immunohistochemical technique was used to localize nerve cell bodies and nerve fibres in the stomach, small intestine and colon of the guinea-pig and the mouse small intestine. Reactive nerve cell bodies were found in both the myenteric and submucous ganglia. Varicose nerve fibres were in the ganglia, in the circular smooth muscle and in the mucosa of the small intestine.

Animals↗
PubMed1983

Application of a modified 203Hg binding assay for metallothionein.

A sensitive and rapid method to estimate concentrations of functional metallothionein in small biological samples, based upon the acid stability of 203Hg binding and solubility of this protein in trichloroacetic acid is described. Sephadex G-10 minicolumns supported in centrifuge tubes afforded separation and quantitation of isotope bound metallothionein from unbound metal. Elution of metallothionein bound 203Hg was achieved by short term-low speed centrifugation that segregated chelator-ligand complex into the eluate while unbound ligand remained in the gel. A well characterized standard of pure metallothionein protein was utilized to verify the specificity and sensitivity of the modified assay. Metallothionein levels were estimated by 203Hg binding in extracts of wild type and cadmium resistant Chinese hamster ovary cells treated with maximum tolerable concentrations of CdCl2. Similar separation methods demonstrated [35S]-cysteine incorporation into induced metallothionein. Additionally, induction of metallothionein was observed after treatment with particulate CdS but not crystalline NiS particles. These results demonstrate that the modified assay system is easily applied to serial measurement of metallothionein levels in multiple small biological samples.

Animals↗
PubMed1983

Distribution, pathways and reactions to drug treatment of nerves with neuropeptide Y- and pancreatic polypeptide-like immunoreactivity in the guinea-pig digestive tract.

Pancreatic polypeptide-like immunoreactivity (PPLI) has been localized in nerves of the guinea-pig stomach and intestine with the use of antibodies raised against avian, bovine and human pancreatic polypeptide (PP), the C-terminal hexapeptide of mammalian PP, and against the related peptide, NPY. Each of the antibodies revealed the same population of neurones. Reactive cell bodies were found in both myenteric (5% of all neurones) and submucous ganglia (26% of all neurones) of the small intestine, and varicose processes were observed in the myenteric plexus, circular muscle, mucosa and around arterioles. The nerves were unaffected by bilateral subdiaphragmatic truncal vagotomy, but the staining of the periarterial nerves disappeared after treatment of animals with reserpine or 6-hydroxydopamine and was also absent after mesenteric nerves had been cut and allowed to degenerate. Vascular nerves showing immunoreactivity for dopamine beta-hydroxylase and PPLI had the same distribution. It is concluded that PPLI is located in periarterial noradrenergic nerves. However, other noradrenergic nerves in the intestine do not show PPLI, and PPLI also occurs in nerves that are not noradrenergic. Analysis of changes in the distribution of terminals after microsurgical lesions of pathways in the small intestine showed that processes of myenteric PP-nerve cells provide terminals in the underlying circular muscle and in myenteric ganglia up to about 2 mm more anal. Submucous PP-cell bodies provide terminals to the mucosa.

Animals↗
PubMed1983

Differential effects of amorphous and crystalline nickel sulfide on murine alpha/beta interferon production.

Mouse embryo fibroblast cultures were exposed to various concentrations of amorphous or crystalline NiS (nickel sulfide). After removal of the NiS from the cultures, polyriboinosinic-polyribocytidylic acid was added to induce alpha/beta interferon. Pretreatment of the cultures with crystalline NiS significantly inhibited the production of alpha/beta interferon at relatively noncytotoxic concentrations (i.e., 1 microgram/ml) while amorphous NiS had no significant effect on alpha/beta interferon production at 10 micrograms/ml. Treatment of cells with reduced glutathione did not block the inhibitory effects of crystalline NiS on alpha/beta interferon production, suggesting that these effects are not mediated by a free-radical mechanism.

Animals↗
PubMed1983

Soluble and insoluble nickel compounds induce DNA repair synthesis in cultured mammalian cells.

The induction of DNA repair was investigated in cultured Syrian hamster embryo (SHE) cells and Chinese hamster ovary (CHO) cells by cesium chloride equilibrium gradient sedimentation techniques following exposure to NiCl2, amorphous NiS, crystalline NiS and crystalline Ni3S2. Significant repair was induced in CHO cells by 1 microgram/ml of crystalline NiS following 24 h of treatment while 5 micrograms/ml caused more than twice the repair activity. In contrast amorphous NiS at 10 micrograms/ml for 24 h induced little repair in these cells. Similarly amorphous NiS did not induce repair at 5-10 micrograms/ml for 24 h in SHO cells while crystalline Ni3S2, and NiCl2 caused substantial induction of DNA repair synthesis at 10 micrograms/ml or 100 microM, respectively. These results demonstrate that nickel compounds which are potent transforming agents and induce damage to DNA also result in the induction of DNA repair. Repair synthesis was detected at concentrations of metal compounds which result in no detectable damage to DNA.

Animals↗
PubMed1983

Distribution and projections of nerves with enkephalin-like immunoreactivity in the guinea-pig small intestine.

Whole mounts of guinea-pig small intestine were used to examine the distribution of neurons with enkephalin-like immunoreactivity and the effects of microsurgical lesions on these neurons. The enkephalin neurons are intrinsic to the intestine. Cell bodies are found in the myenteric ganglia; processes are in the myenteric plexus, circular muscle (including deep muscular plexus) and submucosa, but not in the mucosa. The cell bodies have one prominent process and several short processes, the latter occasionally are seen to give rise in turn to fine, faint processes. The prominent processes provide fibres to the circular muscle and deep muscular plexus beneath and just anal (up to about 2 mm) to the cell bodies. Fibres in the submucous ganglia come from the overlying myenteric plexus. Orally-directed processes (possibly dendrites) of myenteric cell bodies provide the varicose fibres in the myenteric ganglia. These processes are 3.5-4 mm long. The enkephalin neurons represent a population of enteric neurons, with a distinct distribution and projections, which does not correspond to any of the other populations of enteric neurons that have been studied.

Animals↗
PubMed1983

The origins, pathways and terminations of neurons with VIP-like immunoreactivity in the guinea-pig small intestine.

We have analyzed changes in the distributions of terminals with vasoactive intestinal polypeptide (VIP)-like immunoreactivity, and accumulations in severed processes, that occur after lesions of intrinsic and extrinsic nerve pathways of the guinea-pig small intestine. The observations indicate that enteric vasoactive intestinal polypeptide immunoreactive neurons have the following projections. Nerve cell bodies in the myenteric plexus provide varicose processes to the underlying circular muscle; the majority of these pathways, if they extend at all in the anal or oral directions, do so for distances of less than 1 mm. Nerve cell bodies of the myenteric plexus also project anally to provide terminals to other myenteric ganglia. The lengths of the majority of these projections are between 2 and 10 mm, with an average length of about 6 mm. Processes of myenteric neurons also run anally in the myenteric plexus and then penetrate the circular muscle to provide varicose processes in the submucous ganglia at distances of up to 15 mm, the average length being 9-12 mm. In addition, there is an intestinofugal projection of myenteric neurons whose processes end around nerve cell bodies of the coeliac ganglia. A similar projection from the colon supplies the inferior mesenteric ganglia. The nerve cell bodies in submucous ganglia give rise to a subepithelial network of fibres in the mucosa and also supply terminals to submucous arterioles. It is concluded that vasoactive intestinal polypeptide is contained in neurons of a number of intrinsic nerve pathways, influencing motility, blood flow and mucosal transport. The myenteric neurons that project to prevertebral sympathetic ganglia may be involved in intestino-intestinal reflexes.

Animals↗
PubMed1983

Vasoactive intestinal peptide-like immunoreactivity in nerves associated with the cardiovascular system of guinea-pigs.

The distribution of nerves with vasoactive intestinal peptide (VIP)-like immunoreactivity has been examined in the heart and vascular system of guinea-pigs. There was a very sparse supply of fibres to the heart. No immunoreactive cell bodies were found in the intrinsic cardiac ganglia; however, positive nerve cell bodies were seen along the superior vena cava near the right atrium. There were immunoreactive fibres with most arteries; these fibres were located at the media-adventitia junction. The supply to major distributing arteries, such as the aorta, subclavian, carotid and femoral arteries as well as to the pulmonary arteries, was sparse. Of the individual vascular beds, the most densely supplied arteries were the mesenteric and uterine (or in the male deferential) arteries. Arteries running to other organs or tissues, such as skeletal muscle, kidney, pancreas, spleen and heart were less densely supplied. There were clear differences in the innervation of different cerebral vessels. The greatest density was associated with the anterior and middle cerebral arteries. Fewer nerves accompanied the posterior cerebral, cerebellar and meningeal arteries. There was a sparse innervation of the rostral part of the basilar artery. Throughout the body, veins were sparsely supplied. The distribution of nerves with VIP-like immunoreactivity was not changed when noradrenergic nerves were degenerated by 6-hydroxydopamine or when substance P nerves were disrupted by capsaicin. It is concluded that VIP containing nerves innervating the heart and blood vessels form a population distinct from the substance P-containing and the noradrenergic nerves. It is suggested that the VIP fibres might be efferent vasodilator nerves to the blood vessels.

Animals↗
PubMed1983

Vasoactive intestinal polypeptide immunoreactive nerve fibres in the human eye.

Immunohistochemistry applied to whole-mount preparations was used to investigate the presence and distribution of vasoactive intestinal polypeptide (VIP) immunoreactive nerves in the non-retinal part of the human eye. The choroid has a dense perivascular supply of VIP immunoreactive nerve fibers, and some free nerve endings within the stroma. These nerves enter the choroid in ciliary nerves and also as perivascular networks around the ciliary arteries. Occasional choroidal VIP immunoreactive nerve cell bodies are seen. The ciliary body stroma, close to the iris root has a dense circumferential plexus of VIP immunoreactive nerve fibers that occur both singly and in bundles. The iris root has a circumferential arrangement of bundles from which VIP immunoreactive nerve fibres travel radially in the stroma. They supply the pupillary region with numerous free nerve endings; the sphincter pupillae is not supplied by these nerves. The cornea is devoid of VIP immunoreactive nerves. These findings, together with existing knowledge of the physiological actions of VIP, indicate that VIP immunoreactive nerves are likely to be involved in the functioning of several ocular tissues.

Choroid↗