Biomedical subjects

M H Sullivan

Publications and source records attributed to M H Sullivan.

At least 19 recordsLinked to original sources

PubMed2004

Impaired retrograde transmission of vasodilatory signals via the endothelium in pre-eclampsia: a cause of reduced tissue blood flow?

There is evidence that tissue blood flow is regulated by retrograde transmission of signals initiated at capillary and post-capillary sites, and transmitted via the endothelium to modulate pre-capillary resistance. We have used pre-eclampsia as a model to test the hypothesis that normal endothelium is required to enable adjustment of blood flow to match tissue requirements. Integrity of the endothelial pathway was assessed by measuring calf blood flow at increasing venous pressures, using an established small cumulative-step venous-congestion plethysmography protocol in ten women with pre-eclampsia, 17 normal pregnant controls and ten non-pregnant women. Endothelial cell activation was assessed by measuring plasma levels of the cell adhesion molecules, intercellular cell-adhesion molecule-1 (ICAM-1), vascular cell-adhesion molecule-1 (VCAM-1) and E-selectin. Baseline calf blood flow was significantly lower in pre-eclampsia than in the other two groups (P<0.0001; ANOVA). In the pre-eclampsia group, there was a fall in blood flow as venous congestion pressure was raised (P<0.0001; ANOVA). No such change was observed in the other two groups. A significant inverse correlation was observed between the reduction in blood flow in pre-eclampsia and the levels of E-selectin (r=-0.92, P=0.0002), VCAM-1 (r=-0.93, P=0.0008) and ICAM-1 (r=-0.86, P=0.001). The differences between the pre-eclamptic women and the other two groups support the notion that the failure to sustain blood flow during a cumulative pressure step protocol in the pre-eclamptic group might be influenced by interference with the retrograde transmission of signals via the endothelium in these patients.

Adult↗

PubMed2002

Complexities in ETS-domain transcription factor function and regulation: lessons from the TCF (ternary complex factor) subfamily. The Colworth Medal Lecture.

The ETS-domain transcription factor family can be divided into a series of subfamilies. Elk-1 represents the founding member of the ternary complex factor (TCF) subfamily. By focusing on the TCF subfamily, we can demonstrate the complexities that exist in the function and regulation of ETS-domain transcription factors. This article focuses on Elk-1 in detail and summarizes the functions of other TCFs. The key themes covered include the domain structure of the TCFs, the mechanisms of complex formation with serum response factor, regulation of TCFs by mitogen-activated protein kinase cascades, and transcriptional regulatory properties of the TCFs. Finally, the emerging role of the TCFs in vivo is discussed. A picture is developing indicating that, while these proteins exhibit significant sequence and functional conservation, key differences in their structure and regulation are being identified which may relate to unique functions of these proteins in vivo.

Amino Acid Sequence↗

PubMed2001

Intrauterine T-cell activation and increased proinflammatory cytokine concentrations in preterm infants with cerebral lesions.

Brain injury is common in very preterm infants, and intrauterine infection is a frequent antecedent of preterm birth. We examined the relation of cerebral damage to intrauterine antigen exposure and inflammation in 50 infants who were born at 23-29 weeks' gestation. Higher concentrations of cytokines (tumour necrosis factor alpha [TNF-alpha], and interleukins [IL], 1beta, 6, and 10) and CD45RO(+) T lymphocytes in umbilical blood predicted cerebral lesions detected by magnetic resonance imaging very soon after delivery. Our results suggest that infants who mount an immune response in utero are at higher risk of cerebral lesions.

Brain Injuries↗

PubMed2001

Localisation of mRNAs for diamine oxidase and histamine receptors H1 and H2, at the feto-maternal interface of human pregnancy.

OBJECTIVE AND DESIGN: To localise mRNAs for the histamine receptors H1, H2 and H3, and for diamine oxidase, in the placenta and decidua of the human feto-maternal interface. MATERIALS AND METHODS: Complementary DNA for each mRNA of interest was amplified by polymerase chain reaction. Sub-cloned sequences were used to prepare probes for in situ hybridisation, and these were employed to localise the expression of mRNAs for histamine receptors H1 and H2, and for diamine oxidase. RESULTS: mRNA for histamine receptors H1 and H2, and for diamine oxidase could be detected at the feto-maternal interface of human pregnancy, and localised to both decidual and placental cells. CONCLUSION: The co-expression of these receptors and DAO is consistent with a role for histamine at the feto-maternal interface of human pregnancy.

Amine Oxidase (Copper-Containing)↗

PubMed2001

IL-1beta and IL-8 in human fetal membranes: changes with gestational age, labor, and culture conditions.

PROBLEM: Interleukin (IL)-1beta and IL-8 are associated with labor. This study aimed to characterize their concentrations in fetal membranes and any changes in these with advancing gestation and to define as to whether there are interactions between the membranes in their expression. METHOD OF STUDY: mRNA and protein content of amnion and choriodecidua at increasing gestations and before and after labor at term were quantified. Membranes were also collected before and after labor, separated, and cultured. Protein production was measured by ELISA. RESULTS: IL-1beta and IL-8 concentration increased in third trimester amnion and choriodecidua. Further increased expression of mRNA of both cytokines was found after labor in both membranes except IL-8 production by amnion. Choriodecidua produced more of each cytokine than amnion, however, no interaction between the membranes was demonstrated by culture. CONCLUSIONS: Increasing expression of IL-1beta and IL-8 in amnion and choriodecidua in the third trimester and after labor supports a role for these cytokines in the establishment of labor.

Amnion↗

PubMed2001

Changes in the effects of interleukin-1beta and tumor necrosis factor-alpha on platelet activation in early pregnancy.

Platelets are known to be activated in normal pregnancy, and are further activated in pathological pregnancy states, such as preeclampsia. The factors controlling platelet activation are unknown, but cytokines, such as interleukin 1beta (IL-1beta) and tumor necrosis-alpha (TNF-alpha) have been found to affect platelet function and are believed to be involved in early pregnancy. We assessed the effects of these cytokines on platelets from women at various stages of pregnancy. We compared two methods: platelet in vitro aggregation by aggregometry, and platelet P-selectin expression by flow cytometry. IL-1beta and TNF-alpha had no effect on the in vitro aggregation and P-selectin expression of platelets from women in the first trimester of pregnancy as compared to the inhibitory effects of both in late pregnancy. We conclude that maternal platelet function undergoes a marked change throughout pregnancy.

Female↗

PubMed2000

Blood eosinophils from atopic donors express messenger RNA for the alpha, beta, and gamma subunits of the high-affinity IgE receptor (Fc epsilon RI) and intracellular, but not cell surface, alpha subunit protein.

BACKGROUND: Blood eosinophils from hypereosinophilic donors were previously reported to possess the functional high-affinity IgE receptor (Fc epsilon RI), so providing a potential mechanism to account for eosinophil degranulation in atopic allergic disease. Furthermore, tissue eosinophils from allergic tissue reactions were shown to be mRNA(+) for the alpha, beta, and gamma subunits of Fc epsilon RI and gave positive immunostaining with an anti-Fc epsilon RI-alpha antibody. Recent studies, however, revealed negative surface staining on peripheral blood eosinophils, but intracellular Fc epsilon RI-alpha protein was identified by Western blot analysis. OBJECTIVE: Our purpose was to examine on peripheral blood eosinophils from atopic subjects (1) surface expression and mRNA for Fc epsilon RI-alpha, (2) up-regulation of Fc epsilon RI-alpha by allergy-associated tissue factors, and (3) Fc epsilon RI-alpha-dependent release of eosinophil peroxidase (EPO). METHODS: We measured (1) Fc epsilon RI mRNA expression by in situ hybridization, (2) Fc epsilon RI-alpha by flow cytometry and immunocytochemistry (with use of nonpermeabilized and permeabilized cells), and (3) Fc epsilon RI-alpha-dependent release of EPO. RESULTS: Eosinophils from atopic donors had negligible surface expression of Fc epsilon RI-alpha, which was not enhanced by culture with IgE, IL-3, IL-4, IL-5, GM-CSF, or fibronectin or coculture with fibroblasts. Permeabilization, however, revealed appreciable intracellular staining for Fc epsilon RI-alpha. The majority of eosinophils were mRNA(+) for the alpha, beta, and gamma subunits of Fc epsilon RI. Small but significant (P =.03) increases in alpha chain mRNA expression were observed after coculture of eosinophils with fibroblasts but not with IgE, IL-4, or fibronectin. Cross-linking of Fc epsilon RI on the surface of eosinophils from atopic donors did not lead to detectable EPO release. CONCLUSION: Human blood eosinophils express negligible, nonfunctional membrane Fc epsilon RI-alpha but have intracellular Fc epsilon RI-alpha protein and mRNA expression for the alpha, beta, and gamma subunits.

Cell Membrane↗

PubMed2000

The regulation of prostaglandin output from term intact fetal membranes by anti-inflammatory cytokines.

Prostaglandins are some of the main mediators which control parturition, and their production by intrauterine tissues can be up-regulated by pro-inflammatory cytokines. Anti-inflammatory cytokines may oppose these effects, and in this study we have investigated how two such cytokines affected fetal membrane function. Interleukin-10 (IL-10) inhibited the output of prostaglandin E2 (PGE2) from intact fetal membranes under basal and lipopolysaccharide (LPS)-stimulated conditions, and there was a parallel decrease in the expression of mRNA for COX-2. IL-10 also inhibited the production of interleukin-1beta (IL-1beta) and the expression of mRNA for IL-1beta, indicating that this cytokine has a broad anti-inflammatory effect. Transforming growth factor-beta1 (TGF-beta1), which is generally considered to be anti-inflammatory had opposite effects on PGE2 production, in that it increased the output of PGE2 for up to 8 hr. TGF-beta1 increased levels of type-2 cyclo-oxygenase (COX-2) and cytosolic phospholipase A2 (cPLA2) protein, and also activated the cPLA2 enzyme present; the profile of effects is similar to that of the pro-inflammatory cytokine IL-1beta, and was not expected. Combinations of TGF-beta1 with IL-1beta also increased PGE2 output and caused appropriate changes in prostaglandin pathway enzymes, whereas TGF-beta1 and IL-1alpha had more limited effects. Further studies are needed to establish the physiological significance of these findings, but TGF-beta1 does not seem to act as an inhibitory cytokine in intact fetal membranes at term.

Cell Membrane↗

PubMed2000

The roles of the cyclo-oxygenases types one and two in prostaglandin synthesis in human fetal membranes at term.

UNLABELLED: The aim of this study was to determine the relative contributions of cyclo-oxygenase (COX) types 1 and 2 to prostaglandin synthesis at term. METHODS: Fetal membranes were collected from 6 pregnancies after elective caesarean section at term, prior to labour. The presence of COX-1 and COX-2 protein was determined using Western analysis. The relative contributions of the two isoforms of COX to prostaglandin synthesis were determined by incubation of fetal membrane discs with either a COX-2 selective inhibitor, SC236, or a COX-1 selective inhibitor, SC560, and measurement of prostaglandin release during 24 h using enzyme-linked immuno-sorbent assay (ELISA). RESULTS: Both COX-1 and COX-2 protein were demonstrated in amnion and chorion-decidua. The COX-2 selective inhibitor, SC-236, significantly reduced prostaglandin synthesis, both in its COX-2 specific and higher, non-specific concentration ranges. The COX-1 selective inhibitor, SC-560, had no effect upon prostaglandin synthesis in its COX-1 specific concentration range, but did significantly reduce prostaglandin synthesis at higher, non-selective concentrations. CONCLUSIONS: Fetal membranes contain both COX-1 and COX-2 at term, but only COX-2 contributes towards prostaglandin synthesis. COX-2 selective NSAI drugs will be as effective as non-selective agents in inhibition of fetal membrane prostaglandin synthesis and may represent a new strategy for tocolysis.

Cyclooxygenase 1↗

PubMed1999

The production of interleukin-1beta from human fetal membranes is not obligatory for increased prostaglandin output.

Bacterial endotoxin increased the expression of mRNA (maximal after 4 hr) for interleukin-1beta (IL-1beta) and the release of mature protein from intact human fetal membranes. In contrast, the change in expression of mRNA for type 2 cyclo-oxygenase (COX-2) was biphasic, with peaks after 0.5-1 hr and after 8 hr of culture. An antibody to IL-1beta was without effect after 4 hr of culture, inhibited endotoxin-stimulated prostaglandin E2 (PGE2) production after 8 hr of culture, and caused a parallel decrease in the expression of mRNA for COX-2. We conclude that endotoxin induced the expression of COX-2 through IL-1beta-independent and IL-1beta-dependent mechanisms, and these differences are time dependent. Corticotrophin-releasing hormone (CRH) or platelet-activating factor (PAF) also increased the expression of mRNA for IL-1beta and the release of IL-1beta from some, but not all, fetal membranes. The antibody to IL-1beta did not affect CRH-stimulated or PAF-stimulated PGE2 production or COX-2 expression. We conclude that CRH and PAF can induce the expression of IL-1beta, but this is not obligatory for increased PGE2 release, and the effect of these stimuli on COX-2 expression is a direct, IL-1beta-independent effect.

Corticotropin-Releasing Hormone↗

PubMed1999

Corticotrophin-releasing hormone and platelet-activating factor induce transcription of the type-2 cyclo-oxygenase gene in human fetal membranes.

Corticotrophin-releasing hormone (CRH) and platelet-activating factor (PAF) are considered to be involved in the physiological processes of human labour. Both may have dual effects, directly regulating myometrial contractility and fetal membrane prostaglandin production. During this study, we investigated the mechanisms through which CRH and PAF exert their latter effect. CRH and PAF increased prostaglandin production from intact fetal membrane discs, with a maximum stimulation after 8 h of culture. Reverse transcription-polymerase chain reaction (RT-PCR) analyses using primers specific for type-2 cyclo-oxygenase (COX-2) showed that CRH and PAF increased the transcription of COX-2 mRNA two-fold after 8 h culture. These data indicate that the increased fetal membrane prostaglandin production in response to CRH or PAF may involve the induction of COX-2.

Corticotropin-Releasing Hormone↗

PubMed1999

Expression of 5-lipoxygenase and 5-lipoxygenase-activating protein in human fetal membranes throughout pregnancy and at term.

Lipoxygenase metabolites may be involved in human parturition. 5-lipoxygenase (5-LOX) catalyses the first steps in the synthesis of leukotrienes from arachidonic acid, and its activity is dependent on 5-LOX activating protein (FLAP). The expression of 5-LOX and FLAP were investigated in fetal membranes to determine whether there are changes with gestational age or at term with the onset of labour. No significant differences were found in the expression of 5-LOX or FLAP mRNA in the amnion at different gestational ages or at term. In the chorion-decidua, 5-LOX mRNA expression was significantly higher in the first trimester of pregnancy than in the second and third trimesters. At term, there was a significant increase in both 5-LOX mRNA and protein expression in the chorion-decidua in the time after labour, compared with the time before labour. The expression of FLAP mRNA was also significantly higher in the chorion-decidua in the first trimester of pregnancy compared with the third trimester, and at term in the time after labour compared with the time before labour. Expression of FLAP protein was not studied, as an antibody is not currently available. These results are consistent with a role for 5-LOX and FLAP in the control of parturition at term, and also suggest an involvement earlier in pregnancy.

5-Lipoxygenase-Activating Proteins↗

PubMed1999

An inhibitor of type-1 cyclo-oxygenase in tissues from human pregnancy.

BACKGROUND: Previous work has shown that first trimester human decidua contains a protein which directly inhibits the activity of type-1 cyclo-oxygenase (COX-1). METHODS: Activity assays for cyclo-oxygenase types I and II were developed. Cell cytosol was prepared from a number of different sources: human placenta and decidua (first and third trimester), two placental cell-lines (BeWo and TCL-1), an endometrial stromal cell-line and K562 erythroleukemia cells. The effects of all cytosols on activity of type I cyclo-oxygenase, and of cytosols from BeWo choriocarcinoma and decidual cells on type II enzyme, were tested. RESULTS: Cytosols from first trimester human placenta, two placental cell-lines, an endometrial stromal cell-line and K562 erythroleukemia cells all inhibited the type I enzyme. The inhibitor protein could not be detected in third trimester human decidual cells after labor, and was present only at very low levels in third trimester decidua prior to the onset of labor. Cytosols from BeWo and decidual cells had no effect on the activity of the type-2 cyclo-oxygenase enzyme. CONCLUSIONS: The inhibitor of type I cyclo-oxygenase was not specific to pregnancy-related tissues, and may be a general regulator of this enzyme. Lower levels of inhibitor were present at term, but the physiological significance of this is unclear. The cytosolic inhibitor appears to be specific to the type I enzyme.

Cell Line↗

PubMed1998

Interleukin-1beta and bacterial endotoxin change the metabolism of prostaglandins E2 and F2alpha in intact term fetal membranes.

There is strong evidence that prostaglandins E2 and F2alpha (PGE2 and PGF2alpha) are involved in the initiation and maintenance of human parturition and that their production can be stimulated by a number of cytokines and in infection-induced preterm labour by bacterial endotoxin. This study used an intact fetal membrane disk model to investigate the regulation of PGE2 and PGF2alpha metabolism by interleukin-1 beta (IL-1beta) and bacterial endotoxin [lipopolysaccharide (LPS)]. Fetal membrane explants were incubated with IL-1beta (0.1 or 1.0 ng/ml) or LPS (10 ng/ml) for 24 h. A mixture of 3H-prostaglandin (0.1 microCi) and unlabelled prostaglandin (1 microg) was then added at selected times after the addition of inflammatory mediators. The radiolabelled prostaglandins and their metabolites were then extracted from the culture medium and quantified by high-pressure liquid chromatography. Levels of prostaglandin metabolites were generally decreased following incubation with IL-1beta or LPS, which is consistent with a decrease in the activity of 15-hydroxyprostaglandin dehydrogenase (PGDH). It is concluded that IL-1beta and LPS moderately decrease the metabolism of prostaglandins, which may contribute to increasing the local levels of active prostaglandins induced by these stimuli.

Adult↗

PubMed1998

Stimulation of prostaglandin production from intact human fetal membranes by bacteria and bacterial products.

The addition of live or sonicated Escherichia coli, or endotoxin from E. coli increased the release of prostaglandins (PG) on both sides of intact human fetal membranes after 24 h of incubation, indicating that live bacteria were not required to activate prostaglandin production. Time-course studies showed that the levels of PGE2 and PGF2alpha on the fetal side of the membrane were increased 6 h after the addition of endotoxin, whereas levels on the maternal side increased within 1-2 h. These changes were independent of the side to which the endotoxin was added, indicating that a stimulatory factor passes through the fetal membranes. This factor is not endotoxin, which did not cross the membranes, and further studies are required to identify this endogenous stimulus. Prostaglandin metabolite levels were either unaffected or increased by endotoxin, indicating that the main effect is at the level of increased prostaglandin biosynthesis rather than decreased metabolism.

Dinoprost↗

PubMed1998

A spontaneous induction of fetal membrane prostaglandin production precedes clinical labour.

Fetal membranes from term human pregnancies produce prostaglandins, and may respond to bacterial endotoxin or interleukin-1 beta (IL-1 beta) with increased prostaglandin E2 (PGE2) production. The effects of endotoxin persisted for up to 24 h, whereas those of IL-1 beta were maximal 4-8 h after addition. The maximum levels of PGE2 (200-350 pg/ml) were similar in all experiments, and were independent of the stimulus used. Not all tissues responded to these stimuli; those which did not had basal levels of PGE2 production of 200-350 pg/ml, which was not further increased by endotoxin or IL-1 beta. The basal production from these tissues was therefore similar to the maximal production from those tissues which responded to endotoxin or IL-1 beta. The high basal production of PGE2 was attributed to prior in vivo activation of the membranes such that PGE2 synthesis could not be further stimulated in vitro. Overnight pretreatment with aspirin decreased basal PGE2 production from these activated membranes to < 100 pg/ml/4 h during subsequent culture in aspirin-free medium. Both endotoxin and IL-1 beta increased PGE2 production from the activated aspirin-pretreated membranes during this culture time, but this was transient as after 12 h of culture basal PGE2 production rose to over 200 pg/ml despite aspirin pretreatment.

Analysis of Variance↗

PubMed1998

Regulation of prostaglandin production in intact fetal membranes by interleukin-1 and its receptor antagonist.

There is strong evidence for the involvement of inflammatory mediators such as interleukin (IL)-1 in the biochemical mechanisms of parturition. Therefore the effects of the IL-1 family (IL-1alpha (1 ng/ml), IL-1beta (1 ng/ml) and the IL-1 receptor antagonist (IL-1ra) (10 ng/ml)) on the regulation of prostaglandin synthesis in term human fetal membranes were investigated. It was found that, after 4 h of culture, IL-1beta increased prostaglandin E2 (PGE2) output approximately twofold. This was associated with both a significant increase in cyclo-oxygenase-2 (COX-2) mRNA levels (approximately fourfold compared with control) and translocation of cytoplasmic phospholipase A2 (cPLA2) from the cytosol to the membrane fraction. IL-1alpha was less effective than IL-1beta at stimulating PGE2 production through similar mechanisms. IL-1ra had no effect on PGE2 output. However, in combination treatments, IL-1ra did not inhibit IL-1alpha- or IL-1beta-stimulated PGE2 output, and increased PGE2 production further compared with IL-1beta alone. IL-1ra decreased IL-1beta-induced COX-2 mRNA expression by about half and significantly increased cPLA2 protein levels, as detected by immunoblotting, when used alone and together with IL-1beta. These results suggest that IL-1ra has partial agonist properties when used together with IL-1alpha and IL-1beta in fetal membranes by increasing cPLA2 protein levels, which leads to an increase in the production of prostaglandins.

Analysis of Variance↗

PubMed1998

Titration of antiplatelet treatment in pregnant women at risk of preeclampsia.

We recruited 111 patients who were considered to be at significantly increased risk of preeclampsia on the basis of previous obstetric history or preexisting medical disorders. All patients were treated with low dose aspirin (75 mg/day) from the first occasion the patient attended the antenatal clinic, regardless of gestational age. If the maternal mean platelet volume (MPV) increased significantly (by > 0.8 fl) from the baseline, antiplatelet treatment was increased. Five pregnancies were lost during the second trimester and 106 of the treated patients had live infants. The incidence of neonatal death (3/106 infants) was much lower than in the previous pregnancies in these patients (32/134 infants). Patients who were treated from the first trimester of pregnancy (group A, 89 patients) did substantially better than those treated from the second trimester (group B, 17 patients) as assessed by the incidence of pre-eclampsia or intrauterine growth restriction (IUGR), gestational age and birthweight at delivery. These data suggest that longitudinal monitoring of the MPV may identify the women who could benefit from increased antiplatelet treatment, and that antiplatelet treatment may be more effective when initiated in the first trimester rather than later in pregnancy.

Abortion, Spontaneous↗