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Biomedical subjects

J A Clements

Publications and source records attributed to J A Clements.

At least 91 records · Page 5Linked to original sources
PubMed1987

Low molecular weight human pulmonary surfactant protein (SP5): isolation, characterization, and cDNA and amino acid sequences.

Pulmonary surfactant is a lipid-protein complex that promotes alveolar stability by lowering the surface tension at the air-fluid interface in the peripheral air spaces. A group of hydrophobic surfactant-associated proteins has been shown to be essential for rapid surface film formation by surfactant phospholipids. We have purified a hydrophobic surfactant protein of approximately 5 kDa that we term SP5 from bronchopulmonary lavage fluid from a patient with alveolar proteinosis and shown that it promotes rapid surface film formation by simple mixtures of phospholipids. We have derived the full amino acid sequence of human SP5 from the nucleotide sequence of cDNAs identified with oligonucleotide probes based on the NH2-terminal sequence of SP5. SP5 isolated from surfactant is a fragment of a much larger precursor protein (21 kDa). The precursor contains an extremely hydrophobic region of 34 amino acids that comprises most of the mature SP5. This hydrophobicity explains the unusual solubility characteristics of SP5 and the fact that it is lipid-associated when isolated from lung.

Amino Acid Sequence↗
PubMed1987

Analgesia from morphine and ketamine. A comparison of infusions of morphine and ketamine for postoperative analgesia.

Ketamine 4 micrograms/kg/minute produced pain relief similar to that from morphine 33 micrograms/minute in a double-blind study that compared analgesia from constant-rate intravenous infusions of the two drugs in 60 patients. The analgesic efficacy of the infusions, as assessed by pain scores and the requirement for supplementary self-administered morphine, was poor. Ventilatory depression, the most significant side effect, occurred only in patients who received morphine infusion. The low dose ketamine infusion did not provide clinically useful analgesia even though adequate plasma concentrations were achieved.

Clinical Trials as Topic↗
PubMed1987

Changes in quantity, composition, and surface activity of alveolar surfactant at birth.

We hypothesized that when the lung makes the transition from the fluid- to the air-filled state at birth, there are changes in physical and functional properties of the alveolar surfactant. To test this hypothesis, newborn rabbits were killed at different times in the first 24 h of life, their lungs lavaged with ice-cold saline, and the lavage fluid subfractionated by differential centrifugation. The phospholipid and protein content and composition and the kinetics of surface tension lowering of the subfractions were examined. We found that with the onset of breathing, shifts occur in the distribution of surfactant subfractions as a surfactant apoprotein-free phospholipid fraction is generated. The ratio of rapidly sedimentable apoprotein-rich to slowly sedimentable, apoprotein-free fractions decreases from 31 at birth to 4 at 24 h of life. Concurrently, rates of surface tension lowering by the subfractions increase with time. The results suggest that the adult pattern of pool sizes and surface activity of alveolar surfactant is not present at birth but evolves slowly over the 1st day of life.

Aging↗
PubMed1987

Metabolism and turnover of lung surfactant.

Despite the grave difficulties that beset researchers who are trying to isolate and characterize the various intracellular and extracellular forms of lung surfactant; despite the serious ambiguities of interpretation of experimental data caused by the multicomponent nature of the surfactant; despite the inaccessibility for diagnostic purposes of this material, which functions deep in the lungs; despite the fact that incisive methods of protein chemistry and molecular biology have only recently been brought to bear on lung surfactant; despite all these problems, our knowledge and understanding of lung surfactant are rapidly increasing. We know now that it is a dynamic system with many parts, and that the reserves of these are normally not large and are recycled. We believe that tight metabolic control must exist to maintain these reserves, and we already have experimental techniques to raise or lower the reserves. Heroic methods of surfactant substitution are now being tested in patients whose deficiency or risk thereof is sufficiently desperate, and the results of these clinical trials, if they are carefully documented with lung function data, will add a new dimension to our knowledge of surfactant dynamics. We may expect in the future that more subtle alterations in surfactant function will become detectable and more sophisticated methods to manipulate the system will be invented. Essential to these will be a deeper understanding of surfactant forms, pathways, kinetics, and controls. Fortunately, research on surfactant structure, metabolism, and turnover intrigues many investigators. They, we may hope, will soon provide the necessary information.

Animals↗
PubMed1987

Lung function in prematurely delivered rabbits treated with a synthetic surfactant.

We treated prematurely delivered rabbit pups with the synthetic surfactant that has been named Exosurf. By weight, Exosurf is 61.8% dipalmitoylphosphatidylcholine, 6.8% hexadecanol, 4.6% tyloxapol, and 26.7% NaCl. This simple mixture, suspended at 15 mg lipid X ml-1 water, has appropriate in vitro characteristics for a lung surfactant substitute. As determined by static pressure volume relationships performed after 30 min ventilation, lungs treated with Exosurf accepted significantly more gas at maximal inflation (36 versus 15 ml X kg-1 body weight) and had significantly greater volumes during deflation that did saline-treated control lungs; lungs treated with natural rabbit surfactant (SAM) had significantly larger volumes at maximal inflation (65 versus 35 ml X kg-1) and during deflation than did the Exosurf-treated lungs. After 30 min of ventilation with oxygen and fixation at 10 cm H2O pressure, the ratio of air space to tissue space was determined by a point-counting technique, and mean linear intercepts were measured for air spaces. Exosurf-treated lungs were intermediate between SAM and saline-treated lungs in both measurements. With positive pressure ventilation to maintain a tidal volume of 6.5 to 7.5 ml X kg-1, total compliance was significantly greater and inspiratory pressure significantly lower in both SAM- and Exosurf-treated animals than in saline-treated control animals, although the lungs of the SAM-treated animals were more compliant than the lungs of animals treated with Exosurf. During the first minute of positive pressure ventilation, lungs treated with SAM or Exosurf expanded equally rapidly, both expanding more rapidly than the saline-treated lungs.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1987

Concomitant dopaminergic and glucocorticoid control of pituitary proopiomelanocortin messenger ribonucleic acid and beta-endorphin levels.

Synthesis and secretion of POMC-derived peptides appear to be differentially regulated in the anterior pituitary (AP) and neurointermediate lobe (NIL). In the AP, glucocorticoids inhibit, and CRF and arginine vasopressin stimulate, synthesis of POMC and release of immunoreactive (ir)-beta-endorphin (beta EP); in the NIL, synthesis and release of POMC and its derivatives are under tonic inhibitory dopaminergic control. There is, however, evidence for some overlap of these control mechanisms under certain circumstances. In the present study we have used specific RIA and Northern blot analysis to examine the effects of chronic treatment with dopaminergic agents and dexamethasone (DM) (both alone and in combination) on AP and NIL content of ir-beta EP and POMC messenger RNA (mRNA), and/or hypothalamic ir-arginine vasopressin and ir-CRF content. In the NIL, the dopamine agonist bromocriptine reduced and the antagonist haloperidol raised both POMC mRNA and ir-beta EP content. Long term DM treatment did not alter NIL ir-beta EP content in the intact rat, but increased levels of POMC mRNA. DM abolished the haloperidol-induced increase in NIL ir-beta EP content but further increased the haloperidol-induced rise in POMC mRNA. DM treatment lowered both ir-beta EP and POMC mRNA in the AP as well as lowering levels of hypothalamic ir-CRF. In DM-treated rats, haloperidol partially restored AP ir-beta EP and POMC mRNA to control untreated levels. These findings further support the proposition that both dopaminergic agents and glucocorticoids can modulate POMC mRNA levels and/or tissue content of ir-beta EP in both the NIL and AP of the rat. The effects of DM on the NIL, both alone or with haloperidol, suggest that glucocorticoids may have both direct and indirect effects on POMC gene expression in this tissue.

Animals↗
PubMed1986

Inhibin A-subunit gene expression in the ovaries of immature female rats is stimulated by pregnant mare serum gonadotrophin.

We have used a bovine cDNA probe for the A-subunit of inhibin to demonstrate the presence of inhibin in the ovaries of immature female rats: the levels of mRNA were increased following treatment of the rats with pregnant mare serum gonadotrophin (PMSG), suggesting modulation by PMSG of ovarian inhibin synthesis at least at the level of transcription. Furthermore, we report the unexpected finding of significant levels of inhibin mRNA in the corpora lutea of mature female rats. No inhibin mRNA was demonstrable in the uterus or placenta.

Age Factors↗
PubMed1986

Expression of the renal kallikrein gene in mineralocorticoid-treated and genetically hypertensive rats.

On the basis of both clinical observations and experimental studies it has been proposed that renal kallikrein is a mineralocorticoid regulated protein. In other studies, changes in renal kallikrein activity have been implicated in the genesis of, and/or response to, hypertension. Using a cloned complementary DNA (cDNA) to rat pancreatic kallikrein (pcXP39) for hybridization histochemistry, and both Northern and dot blot analysis, we studied expression of the kallikrein gene in steroid-treated control animals, and in three strains of genetically hypertensive rats. No differences in renal kallikrein messenger RNA (mRNA) levels were found between adrenalectomized rats and those treated for 5-14 days with 9 alpha-fludrocortisone, corticosterone or dexamethasone, or between hypertensive rats and their appropriate controls. Since mRNA levels appear essentially invariant under such circumstances, the change in renal kallikrein activity/immunoreactivity after chronic mineralocorticoid elevation, or in hypertensive rats, presumably reflects modulation at the post-transcriptional level.

Animals↗
PubMed1986

A comparison of the pharmacokinetics of propranolol in obese and normal volunteers.

The pharmacokinetics of intravenous and oral propranolol have been compared in six obese and six normal subjects matched for age and sex. After intravenous administration there was no difference in plasma clearance but the volume of distribution was greater (V = 339 l vs 198 l) and the half-life was longer (t1/2 = 5.0 h vs 3.0 h) in the obese group. No important difference in the rate of oral absorption was observed. A trend towards higher systemic availability in the obese group (35% vs 27%) was not statistically significant.

Administration, Oral↗
PubMed1986

Uptake of lung surfactant subfractions into lamellar bodies of adult rabbit lungs.

The goals of this investigation were to determine whether subfractions of alveolar surfactant that have different physical and biochemical properties are preferentially taken up from the alveolar air space into lamellar bodies and to correlate the magnitude of the uptake with the properties of the fractions. Radiolabeled subfractions were obtained by differential centrifugation of lavage fluid from rabbits that had been intravenously injected with radioactive palmitate. The subfractions were P (pellet) 3 (1,000 g, 20 min), P4 (60,000 g, 60 min), P5 (100,000 g, 16 h). Subfractions were instilled into the lungs of anesthetized spontaneously breathing adult rabbits, and lavage and lamellar body fractions were isolated at later times. P3 and P4 were taken up to a larger extent than was P5 or liposomes prepared from a P4 lipid extract. The fractions that were preferentially taken up (P3 and P4) contained surfactant apoprotein (APO) 36, tubular myelin, multilamellar vesicles, and were rapidly adsorbed to an air-water interface. P3 also contained APO 10. These results demonstrate that different forms of surfactant are recycled at different rates and suggest that there is specificity in the recycling process.

Animals↗
PubMed1986

Elevated plasma levels of pro-opiomelanocortin-derived peptides in sheep following hypothalamo-pituitary disconnection.

We have studied the control of the pituitary-adrenal axis in ovariectomized sheep following hypothalamo-pituitary disconnection (HPD) by following plasma levels of immunoreactive (ir-) products of pro-opiomelanocortin (POMC). In ovariectomized HPD ewes, gonadotropin levels were below detection limits. In contrast, levels of ir-ACTH were modestly but significantly elevated over those in matched ovariectomized control ewes, though cortisol levels were not significantly different; levels of ir-alpha MSH and ir-beta EP (beta-endorphin) were substantially and significantly raised in HPD compared with control plasma. Size exclusion HPLC showed plasma beta EP/beta LPH (beta-lipotropin) levels to be higher in HPD than control ewes. Dexamethasone administration lowered plasma ir-ACTH but not ir-beta EP; in contrast, bromocriptine lowered ir-beta EP but not ir-ACTH. We interpret these data as evidence (1) that the elevated plasma levels of ir-beta EP and ir-alpha MSH post-HPD reflect the release of the intermediate lobe from tonic inhibitory dopaminergic control and (2) that, unlike the gonadotrope, hypothalamic releasing factors are not required for the maintenance of the corticotrope, or for baseline secretion of ACTH from these cells.

Adrenal Glands↗
PubMed1986

Pro-opiomelanocortin messenger ribonucleic acid and posttranslational processing of beta endorphin in spleen macrophages.

We have previously demonstrated low levels of immunoreactive (ir)-beta-endorphin (beta-EP) and ir-ACTH in a subpopulation of mouse spleen macrophages, which is consistent with an involvement of opioid peptides in modulation of immune responses. Gel chromatography studies suggested the presence of an approximately 3.5,000-molecular weight (mol wt) species, putatively beta-EP, an approximately 11.5,000-mol-wt species, putatively beta-lipotropin, and a higher molecular weight species (putative beta-EP precursor, pro-opiomelanocortin (POMC). In this study we have extended our original findings by demonstrating the presence of messenger RNA for POMC by the use of a complementary DNA probe and Northern blot analysis of extracts of mouse and rat spleen. In addition, using high performance liquid chromatography (HPLC), we have shown that the major endorphin species in mouse spleen macrophages is beta-EP1-31, and that there are smaller amounts of each of the acetylated forms, N-acetyl-beta-EP1-16 (alpha-endorphin), N-acetyl-beta-EP1-17 (gamma-endorphin), N-acetyl-beta-EP1-27, and N-acetyl-beta-EP1-31. We interpret these studies as showing that (a) the spleen is an organ of POMC synthesis and that (b) the predominant COOH-terminal product of macrophage POMC is the opiate-receptor active species beta-EP1-31.

Animals↗
PubMed1986

Estrogen regulation of kallikrein gene expression in the rat anterior pituitary.

Using a rat pancreatic kallikrein cDNA probe (pcXP39), previously shown to hybridize to kallikrein mRNA in a variety of tissues, we have explored the control of kallikrein gene expression in rat anterior pituitary. Intact female rats have substantially higher levels of AP kallikrein mRNA than intact males; male levels are unaffected by castration, whereas female levels fall markedly postovariectomy. Administration of estradiol benzoate to intact male or ovariectomized female rats causes an increase in anterior pituitary levels of kallikrein mRNA. Since the pattern of responsiveness parallels that of PRL, we have studied GH3 cells grown in the presence and absence of estradiol; in neither instance was kallikrein mRNA above detection limits. Parallel changes were seen on Northern blots and by hybridization histochemistry; on emulsion autoradiography of pituitary sections, scattered positive cells were seen, but precise definition was not possible. We conclude that whereas in the submaxillary gland kallikrein gene expression appears androgen dependent and in the kidney is postulated to be mineralocorticoid regulated, in the anterior pituitary expression of the gene is under estrogen control; and that the local role(s) of pituitary kallikrein, whether precursor processing, control of blood flow, or other effects, would, in turn, appear to be modulated by estrogen in vivo.

Animals↗
PubMed1985

Kallikrein gene expression in the rat anterior pituitary.

The report of 'kallikrein-like' activity in the rat neuro-intermediate lobe (N-IL) and its possible involvement in pro-opiomelanocortin processing led us to explore the expression of the kallikrein gene(s) in the pituitary. Using 32P-labelled rat pancreatic kallikrein cDNA, we have shown positive hybridization for rat anterior pituitary poly(A)+ RNA, of identical size on Northern blots (approximately 1.0 kb) to rat kidney poly(A)+ RNA run in parallel. Prior adrenalectomy or ovariectomy decreased the level of kallikrein mRNA seen in the anterior pituitary; total RNA from rat N-IL showed no significant hybridization. On hybridization histochemistry the anterior pituitary was strongly positive, and the neural and intermediate lobes negative. The previously reported kallikrein-like activity in the N-IL is therefore probably due to a non-kallikrein kininogenase; in the anterior pituitary, kallikrein may have a physiological role in limited precursor proteolysis, but lack kininogen activity.

Animals↗
PubMed1985

Effects of a protein-free, synthetic surfactant on survival and pulmonary function in preterm lambs.

We have created a totally synthetic, protein-free surfactant (Exosurf) composed of dipalmitoylphosphatidylcholine, hexadecanol, and tyloxapol. We studied the effects of endotracheal instillation of Exosurf on survival and pulmonary function of preterm lambs delivered at 131 to 133 days gestation (term 148 days). Exosurf treatment was compared with instillation of surface-active material prepared from lung lavages of adult sheep and with no instillation. Lambs were delivered by cesarean section, paralyzed, and mechanically ventilated. The Exosurf group survived longer (80% alive at 11 hours) than did the no instillation group (30% alive at 11 hours) (P less than 0.05). There were no statistically significant differences between the Exosurf and sheep surfactant groups. We conclude that Exosurf, a synthetic surfactant, produces significant improvement in survival and pulmonary function in preterm lambs.

Animals↗
PubMed1985

N-acetyl endorphin in rat spermatogonia and primary spermatocytes.

In previous reports modest levels of beta-endorphin have been found by radioimmunoassay in rat testis, and localized by immunofluorescence to the interstitial cells. We have confirmed these previous reports and extended them by showing that the majority of testicular endorphins are acetylated forms, N-acetyl gamma-endorphin, N-acetyl alpha-endorphin, and N-acetyl beta-endorphin1-27. In addition, N-acetylated endorphins are not found in interstitial cells, but are confined to spermatogonia and primary spermatocytes.

Aging↗
PubMed1985

Localization of arginine vasopressin-neurophysin II messenger ribonucleic acid in the hypothalamus of control and Brattleboro rats by hybridization histochemistry with a synthetic pentadecamer oligonucleotide probe.

By using 32P-labeled oligonucleotide probes for in situ DNA-RNA hybridization, we have defined the distribution of messenger RNA (MRNA) for arginine vasopressin-neurophysin II (AVP-NPII) in the rat hypothalamus. With a specific pentadecamer nucleotide probe, strong specific hybridization is seen in supraoptic, suprachiasmatic, and paraventricular nuclei in both Sprague-Dawley and Brattleboro rats. This finding, of equivalent distribution of AVP-NPII mRNA in Brattleboro and control hypothalami, confirms and extends the recent report of equivalent levels of AVP-NPII mRNA by Northern blot analysis.

Animals↗