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Biomedical subjects

F Lyall

Publications and source records attributed to F Lyall.

At least 55 records · Page 3Linked to original sources
PubMed1990

Rapid activation of the non-toxic basic isoform of phospholipase A2 from Naja mossambica mossambica (spitting cobra) by long-chain fatty acylation.

Purified phospholipase A2 from Naja naja (common Indian cobra) venom, a basic non-toxic isoform and a toxic isoform from Naja mossambica mossambica venom were treated with an equimolar amount of oleoyl imidazolide, a known activator of bee venom phospholipase A2. The ability of the first two enzymes to induce erythrocyte lysis was strongly activated while hydrolysis of dioctanoyl phosphatidyl choline was only weakly activated. The toxic enzyme showed little change in either assay. The susceptible enzyme from Naja mossambica mossambica venom reacted very much more rapidly with oleoyl imidazolide than did the bee venom enzyme and oleic acid was shown to be a relatively weak antagonist. Activation had very little effect on the metal ion dependence of these enzymes. Although activation was a progressive response having the characteristics of a chemical reaction which could not be mimicked by free fatty acids, the adduct did not appear to be stable under acidic conditions. This evidence suggested that primary amino groups were not the target for acylation.

Acylation↗
PubMed1990

Blood pressure, left ventricular mass and intracellular calcium in primary hyperparathyroidism.

1. Blood pressure, left ventricular mass and platelet cytosolic free calcium concentrations were measured in 23 patients with untreated primary hyperparathyroidism, 30 normotensive control subjects and 23 control subjects matched for age, sex and blood pressure. In 12 patients measurements were repeated after parathyroidectomy. 2. Patients with primary hyperparathyroidism had significantly elevated blood pressures (139 +/- 6/86 +/- 3 mmHg, mean +/- SEM) compared with control subjects (125 +/- 2/78 +/- 1 mmHg), but high values persisted after hypercalcaemia was corrected. 3. Despite chronic extracellular hypercalcaemia, intracellular free calcium levels were lower in patients with hyperparathyroidism than in controls matched for age, sex and blood pressure (median concentrations 81.5 nmol/l vs 93 nmol/l, 95% confidence interval 0.1 to 20.1; P less than 0.05) and values tended to increase after parathyroidectomy. 4. Left ventricular mass index was increased in the primary hyperparathyroid group as compared with control subjects matched for age, sex and blood pressure (123 g/m2 vs 100 g/m2, 95% confidence interval -36.1 to -3.1; P = 0.03). Parathyroidectomy resulted in a small reduction of the left ventricular mass index (123.5 g/m2 vs 104 g/m2, 95% confidence interval 46.5 to 2.5; P = 0.1) but no change in blood pressure. 5. Hypertension and left ventricular hypertrophy in primary hyperparathyroidism are associated with relatively low levels of free calcium in platelets.

Adult↗
PubMed1990

Vascular hypertrophy, renin and blood pressure in the young spontaneously hypertensive rat.

1. Cardiovascular reactivity, blood vessel morphology, blood pressure and the activity of the renin-angiotensin system were determined in the 3-week-old spontaneously hypertensive (SHR), Wistar-Kyoto (WKY) and outbred Wistar (WIS) rat. 2. In an isolated perfused mesenteric artery preparation the SHR had a significantly increased maximum response to KCl and noradrenaline (P less than 0.02) compared with the WKY. Using a myograph, vascular structure was measured over a range of resistance arteries and showed a significant correlation between lumen diameter and both media cross-sectional area and thickness, with the regression line for the SHR shifted upwards indicating both increased media area and thickness. This was associated with a slight, but significant, narrowing of the lumen (P less than 0.01) and an increased media/lumen ratio (0.049 +/- 0.01, 0.034 +/- 0.007, 0.036 +/- 0.008 for SHR, WKY and WIS, respectively, means +/- SD P less than 0.001). The SHR had a greater heart/body weight ratio than either the WKY or the WIS (P less than 0.001). 3. Both mesenteric artery and membrane protein content were higher in the SHR, indicating an increase in cell size or number. 4. Plasma renin activity (means +/- SD) was lower in the SHR (1.0 +/- 0.7 pmol of angiotensin I h-1 ml-1) than in the WKY (2.2 +/- 1.2 pmol of angiotensin I h-1 ml-1, P less than 0.001) but not different from that in the WIS (1.2 +/- 0.8 pmol of angiotensin I h-1 ml-1). Mesenteric artery vascular renin concentration was also lower in the SHR (P = 0.06).(ABSTRACT TRUNCATED AT 250 WORDS)

Angiotensin II↗
PubMed1989

Atrial natriuretic peptide vascular receptor: regulation of low-affinity binding by calcium.

1. In the presence of calcium, both high- [dissociation equilibrium constant (Kd) 5.8 x 10(-11) mol/l] and low- (Kd 1.2 x 10(-8) mol/l) affinity binding of atrial natriuretic peptide was detected on plasma membranes prepared from sodium-depleted rats. 2. In contrast, under calcium-free conditions only high-affinity binding (Kd 6.9 x 10(-11) mol/l) was detected. 3. The requirement for calcium for low-affinity binding suggests a different mechanism of action than that for the high-affinity receptor.

Animals↗
PubMed1988

Cultured mesenteric vascular smooth muscle cells express dopamine DA1-receptors.

Incubation of cultured mesenteric vascular smooth muscle cells with dopamine, in the presence of propranolol, caused an increase in cyclic AMP formation in a concentration-dependent manner (Ka apparent 6.8 +/- 0.5 microM). This effect of dopamine was inhibited by the DA1-receptor antagonist SCH 23390 (Ki = 1 nM). These results suggest that cultured mesenteric vascular smooth muscle cells express DA1-receptors linked to adenylate cyclase.

Adenylyl Cyclases↗
PubMed1988

Characterization of specific binding of atrial natriuretic peptide (ANP) to rat PC12 phaeochromocytoma cells.

Specific binding sites for atrial natriuretic peptide have been identified in membrane of the phaeochromocytoma cell line PC12. Scatchard analysis of binding studies revealed a Kd of 794 pM and a density (Bmax) of 254 fmol/mg protein. Hormones unrelated to ANP such as angiotensin II, bradykinin and arginine-8-vasopressin did not complete for the binding sites. Of the ANP-related peptides which competed for the binding sites, the following order of affinity was established; rANP (8-33) greater than rANP (28 amino acid) greater than rat atrial peptide fragment (13-28) greater than a-hANP (28 amino acid) greater than atrial peptide fragment (1-11) greater than atriopeptin I.

Adrenal Gland Neoplasms↗
PubMed1988

Angiotensin II activates Na+-H+ exchange and stimulates growth in cultured vascular smooth muscle cells.

Angiotensin II (Ang II) stimulated the growth of cultured vascular smooth muscle cells in a dose-dependent manner; a small but significant increase being seen at 10(-9) mol/l, whereas 10(-5) mol/l produced a 166% increase compared with control cells. Saralasin blocked the Ang II-mediated increase in cell number. Amiloride-sensitive Na+-H+ exchange was characterized (Km = 18 mmol/l, Vmax = 0.27 pH units/30 s) in cell monolayers using the fluorescent probe biscarboxyethylcarboxy fluorescein. Angiotensin II (10(-7) mol/l) produced a small overall net intracellular alkalinization. Raising intracellular pH via Na+-H+ activation might be one of the ways in which Ang II increases cell growth.

Angiotensin II↗
PubMed1988

Specific binding of atrial natriuretic peptide increases cyclic GMP levels in human astrocytoma cells.

Specific high-affinity binding sites (dissociation constant 100 pmol/l) for atrial natriuretic peptide (ANP) have been identified in the clone D384 derived from the human astrocytoma cell line G-CCM. Unrelated peptides such as angiotensin II, vasopressin and bradykinin did not compete for these sites. Of the atrial natriuretic peptides studied, both the human and rat ANP competed equally, while peptides with either C- or N-terminal residue missing or with no internal -S-S-bond either competed less effectively or did not compete at all. Human ANP stimulated the cells to increase their intracellular level of cyclic GMP in a time- and dose-dependent manner with maximum stimulation being approached but not reached at concentrations of 1 mumol/l. These results support both the notion that ANP has an important functional role within the brain and the concept of neurotransmitter/neuromodulator communication between neurones and glia.

Astrocytoma↗
PubMed1987

Specific binding of atrial natriuretic peptide to rat mesenteric artery: effect of calcium and 5-guanylylimidodiphosphate.

1. High-dissociation equilibrium constant (20 pmol/1, receptor number 17 fmol/mg) and low-dissociation equilibrium constant (10 nmol/1, receptor number 5 nmol/mg) affinity binding sites for atrial natriuretic peptide have been identified in membrane preparations from rat mesenteric artery. 2. Tracer degradation was corrected for mathematically and binding data were analysed by a non-linear computer technique. 3. Non-atrial natriuretic peptide vasoactive hormones, angiotensin II, vasopressin and bradykinin, did not compete for the high-affinity site. Related peptides with either C- or N-terminal amino acids missing still competed, while peptides without an intact disulphide bridge did not compete. 4. Neither the addition nor the removal of calcium affected the affinity or density of binding sites. Also the non-hydrolysable guanosine triphosphate analogue 5-guanylylimidodiphosphate had no effect on either affinity or number of sites. 5. These results indicate the presence of a specific high-affinity vascular receptor for atrial natriuretic peptide which could interact with the hormone under physiological conditions. The mechanism of binding is uncertain but is unlikely to involve calcium- or guanosine triphosphate-associated regulatory sites.

Animals↗
PubMed1987

Rat atrial natriuretic peptide vascular receptor: effect of alterations in sodium balance and of DOC hypertension.

The effect of changes in dietary sodium intake and of DOC hypertension on plasma atrial natriuretic peptide (PANP), and affinity (Kd) and number (Bmax) of vascular atrial natriuretic peptide binding sites was studied in the rat. There was no difference in PANP between rats on a high or low sodium intake [33.2 +/- 13.9 versus 30.7 +/- 17.3 (s.d.) fmol/ml], Kd [21.1 +/- 2.7 versus 19.7 +/- 4.5 (s.d.) pmol/l] or Bmax [14.8 +/- 1.6 versus 12.6 +/- 1.8 (s.d.) fmol/mg], respectively. In DOC hypertensive rats, PANP was increased compared with control animals [66.1 +/- 32.4 versus 26.4 +/- 9.9 (s.d.) fmol/ml, P less than 0.05] and there was apparent receptor down-regulation [Bmax 7.7 +/- 1.6 versus 19.7 +/- 3.5 (s.d.) fmol/mg, P less than 0.05] with no change in affinity [Kd 15.6 +/- 3.9 versus 18.3 +/- 3.2 (s.d.) pmol/l]. Down-regulation was confirmed when the membrane-bound enzyme 5'-nucleotidase, rather than protein, was used as an index of receptor number. These results suggest that in the rat, atrial natriuretic peptide (ANP) may be important in regulating cardiovascular homeostasis only following non-physiological alterations in sodium and volume status.

5'-Nucleotidase↗
PubMed1985

Activation of bee venom phospholipase A2 by oleoyl imidazolide produces a thiol- and proteinase-resistant conformation.

Assay methods for bee venom phospholipase A2 are presented which respond to different aspects of enzymic behaviour and which allow basal activity, fatty acid activation and acyl-group activation to be distinguished. The stability of the enzyme to thiols and proteinases is dramatically increased by activation with the selective acylating agent, oleoyl imidazolide. These results support the model of activation by conformation change. Limited-fixation studies indicate that enzyme conformation is determined by interaction with the substrate. The oleoyl-enzyme is partially inactivated by trypsin, but its electrophoretic mobility is unchanged. This protective effect is highly selective and only one other component of the venom is protected against trypsin by oleoyl imidazolide. Combination of trypsin and thiol treatment produces a large fragment of the activated enzyme which could be used for structural studies of the activation site.

Bee Venoms↗
PubMedDate not supplied

Heme oxygenase expression in cultured human trophoblast cells during in vitro differentiation: effects of hypoxia.

Heme oxygenases (HO-1 and HO-2) are responsible for the production of carbon monoxide, a vasodilator. HO is important in controlling placental blood flow and expression can be sensitive to oxygen. We previously reported a reduction in HO-2 expression in placentae obtained from patients with pre-eclampsia or living at high altitude, both associated with placental hypoxia. Thus we hypothesized that HO expression in cultured trophoblasts would be altered by exposure to hypoxia. HO-1 and HO-2 expression was assessed in trophoblast cell cultures following exposure to different oxygen environments. Western blot analyses showed that HO-1 expression in syncytiotrophoblast was significantly lower than in cytotrophoblasts in standard conditions (p < 0.05). There was no difference in HO-1 expression in cytotrophoblasts transferred to 2% O2 for various times. However, exposure of syncytiotrophoblast cultures to hypoxia for 12 h resulted in a significant reduction in HO-1 expression (p < 0.05). HO-2 expression was not affected by exposure to hypoxia in either cytotrophoblast or syncytiotrophoblast cultures. Possible interpretations of these findings are that chronic hypoxia alone is not responsible for reduced HO-2 expression or a much longer exposure to chronic hypoxia (perhaps months) is required. This study also reinforces the complexities of HO regulation by oxygen.

Adult↗
PubMedDate not supplied

Alphafetoprotein and alphafetoprotein receptor expression in the normal human placenta at term.

Alphafetoprotein (AFP) is detectable in maternal serum from around six weeks of gestation and is synthesised by the yolk sac and the fetal liver. The role of the placenta in the transport and possible synthesis of AFP is uncertain. The aim of this study was to investigate placental expression of AFP and the AFP receptor in uncomplicated pregnancies at term. Immunohistochemistry and Western blotting clearly demonstrated the presence of AFP in villous tissue at term. However, evidence of AFP mRNA expression or synthesis of AFP was not found following reverse transcription polymerase chain reaction of total RNA isolated from villous tissue and trophoblast cell cultures. The presence of a cell surface receptor for AFP in placental villous tissue, identified by immunohistochemistry and Western blotting, suggests a possible receptor-mediated mechanism for placental transport of AFP while the patterns of expression of AFP and its receptor may indicate a possible route by which AFP is transported across the placenta between the fetal and maternal circulations. These findings demonstrate that the placenta does not synthesise AFP at term and that the presence of AFP in the placenta is a reflection of transplacental transport of AFP possibly via a receptor-mediated mechanism.

Adult↗
PubMedDate not supplied

Nitric oxide concentrations are increased in the feto-placental circulation in intrauterine growth restriction.

The aim of this study was to measure plasma concentrations of total nitrites, as an index of nitric oxide (NO) synthesis, in the fetal circulation of normal pregnancies and in pregnancies complicated by intrauterine growth restriction. Plasma was prepared from umbilical venous blood collected from 13 placentae from normal pregnancies complicated by intrauterine growth restriction. Plasma NO concentrations were determined using the Greiss reaction by measuring combined oxidation products of NO, plasma nitrite (NO2-) and nitrate (NO3-) after reduction with nitrate reductase. Significantly higher NO2-concentrations were found in umbilical venous plasma in the group complicated by intrauterine growth restriction compared to the control group (65.6 mumol/1, P < 0.001. These results support the hypothesis that increased NO production may be a compensatory response to improve blood flow in the placenta and/or may play a role in limiting platelet adhesion and aggregation.

Adult↗
PubMedDate not supplied

Treatment of endothelial cells with serum from women with preeclampsia: effect on neutrophil adhesion.

OBJECTIVE: The purpose of our study was to determine whether the previously reported neutrophil activation that occurs in the maternal circulation of women with preeclampsia is due to a factor (or factors) in serum that increases neutrophil adhesion to endothelial cells. METHODS: The extent of neutrophil adhesion to endothelial cells incubated with serum from women with preeclampsia (n = 12) was compared with serum from normal, pregnant women matched for maternal age and gestational age at blood sampling (n = 12). Preeclampsia was defined as persistent diastolic blood pressure above 90 mmHg, with proteinuria greater than 0.3 g per 24 hours, in patients who were normotensive before 20 weeks' gestation. The ability of serum (with and without heat inactivation of the complement system) from both groups of patients to stimulate neutrophil adhesion to endothelial monolayers was tested in a 15-minute quantitative assay using fluorescence-labeled neutrophils. The extent of neutrophil adhesion was quantified indirectly from fluorescence counts. RESULTS: No significant differences were found regarding neutrophil-endothelial cell adhesion in response to media alone, serum from women with preeclampsia, and serum from normal, pregnant women. This was also the case when the serum was heat inactivated to destroy the complement system. However, heat inactivated serum produced a significantly greater extent of adhesion compared with serum containing an intact complement system, regardless of whether the patient had preeclampsia. CONCLUSION: This study found no evidence of a factor in serum from women with preeclampsia that could alter neutrophil-endothelial cell adhesion via a direct effect on the endothelium. However, our data suggest that adhesion may be regulated in an inhibitory manner by the complement system.

Adult↗
PubMedDate not supplied

Relationship of cell adhesion molecule expression to endothelium-dependent relaxation in normal pregnancy and pregnancies complicated with preeclampsia or fetal growth restriction.

OBJECTIVES: To determine the degree of endothelium-dependent relaxation in myometrial and omental resistance arteries from normal pregnancies and pregnancies complicated with preeclampsia or fetal growth restriction (FGR) (compromised pregnancy group), and to correlate the results with the endothelial surface expression of cell adhesion molecules (CAMs) in the same vessels. METHODS: Parallel wire myography was used to assess the relaxation of omentum or myometrial vessels obtained from nonpregnant women (n = 3), women with normal pregnancies (n = 11), and women with pregnancies complicated by preeclampsia or fetal growth restriction (n = 10). These resistance vessels were constricted with incremental concentrations of vasopressin (10(-10) mol/L to 3.3 x 10(-8) mol/L) prior to the addition of incremental concentrations of bradykinin (10(-10) mol/L to 3.3 x 10(-6) mol/L). Immunohistochemistry was used to assess the endothelial expression of the CAMs E-selectin, ICAM-1, VCAM-1, and PECAM. RESULTS: A significant reduction in endothelium-dependent relaxation of myometrial vessels was found in the compromised pregnancy group when compared with both the normotensive pregnant group and the nonpregnant group. This reduction was not noted with omental vessels. All vessels in the nonpregnant group, normal pregnant group, and compromised pregnancy group expressed PECAM and ICAM-1 on the endothelium. There was no difference in intensity of immunostaining between the groups. None of the vessels in any of the groups expressed VCAM-1 or E-selectin. CONCLUSIONS: We found no evidence that impaired relaxation responses to bradykinin are linked to altered expression of CAMs in preeclampsia and FGR. These results suggest that increased CAM expression occurs in a vascular bed separate from those investigated in the present study. Possible sites for this would be in the microcirculation of organs such as the kidney.

Adult↗
PubMedDate not supplied

Placental and maternal serum inhibin-A and activin-A levels in Down's syndrome pregnancies.

The objective of this study was to analyse the levels of inhibin-A and activin-A in maternal serum and placental tissue from Down's syndrome (DS) pregnancies. Inhibin-A and activin-A levels were determined by specific immunoassays and individual results were expressed as multiples of the control median (MoM) at the appropriate gestation. Immunohistochemistry was used to localize inhibin alpha and beta(A)-subunits in a selection of placental sections. In DS pregnancies, median inhibin-A levels were found to be significantly elevated to 1.46 MoM (P< 0.05) in placental extracts, and 2.06 MoM (P< 0.0001) in maternal serum, when compared with uncomplicated pregnancies. Median activin-A MoMs were also elevated in placental extracts and maternal serum to 1.62 MoM (P< 0.01), and 1.26 MoM (P< 0.05), respectively. Immunohistochemistry revealed that the alpha subunit of inhibin-A and the beta(A)subunit of inhibin-A and activin-A were mainly localized to the trophoblastic layer of placental villi. Semiquantitative studies of staining intensity revealed a trend towards stronger staining of placental trophoblasts and stroma of DS tissues, although this was statistically significant only for beta(A)subunit staining of trophoblasts (P< 0.05). These results support the hypothesis that maternal serum levels of inhibin-A and activin-A are elevated due to increased production in the placenta, and increased immunostaining of trophoblasts suggests that this may be due to increased production in the trophoblasts.

Activins↗