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Biomedical subjects

J Fletcher

Publications and source records attributed to J Fletcher.

At least 73 records · Page 4Linked to original sources

Mechanisms of spiroplasma genome variation associated with SpV1-like viral DNA inferred from sequence comparisons.

Genomes of Spiroplasma citri strains have rearranged frequently during their evolution, partly due to multiple integrated sequences of spiroplasma viruses. To understand better the role of viral sequences in genome evolution, we examined available nucleotide sequences of viruslike elements in the S. citri chromosome. Comparison of integrated and nonintegrated sequences of spiroplasma virus SpV1-C74 DNA suggested that it is an encapsidated form of the circular transposition intermediate belonging to an insertion sequence (IS3) family member. One SpV1-C74 viral DNA fragment was identified as interrupting the remains of a DNA adenine modification methylase gene. A viral DNA insertion of SpV1-R8A2 B DNA had hallmarks of having suffered an internal deletion by a site-specific recombination system. Homologous recombination likely was responsible for several deletions within viral DNA. A homologous recombination event was inferred between part of a viral DNA insertion and a similar chromosomal sequence. Dispersed sequences from SpV1-like C4 open reading frames (ORFs) were identified as involved in a complex deletion-inversion event. Thus, SpV1-like sequences likely have altered spiroplasma genomes by inserting within active genes, destroying their function, by providing targets for site-specific recombination, by mediating deletions of sequences adjacent to their integration sites, and by providing targets for homologous recombination, leading to inversions.

Amino Acid Sequence↗

Neutrophil function in women with pre-eclampsia.

OBJECTIVE: To investigate the function of neutrophils in normal pregnancy and pre-eclampsia. DESIGN: Baseline levels and activated responses of peripheral blood neutrophils were measured in response to the physiological agonists, n-formyl-met-leu-phe (fMLP) and zymosan activated serum. SAMPLE: Neutrophils of 16 pre-eclamptic, 17 normal pregnant (third trimester) and 15 nonpregnant age-matched control women were calculated. SETTING: Antenatal Clinic, City Hospital, Nottingham. METHODS: Neutrophil superoxide anion production was determined by lucigenin-enhanced chemiluminescence; the release of secondary granule lactoferrin by ELISA; and the expression of cell surface adhesion molecules CD11b, CD18 and L-selectin (CD62L) by flow cytometric analysis. RESULTS: Superoxide anion generation was reduced in the pregnant group compared with nonpregnant controls [fMLP by 51% (P = 0.03) and zymosan activated serum by 56% (P = 0.01)] but pre-eclamptic measurements did not show a similar reduction. There were no differences between the three study groups in the plasma levels of lactoferrin, or in the stimulated expression and release of CD11b and CD18, or lactoferrin and L-selectin. The base-line levels for the production of superoxide anions; the expression of CD11b or CD18 or L-selectin; and the release of lactoferrin showed no significant differences. CONCLUSIONS: Circulating neutrophils in pregnancy and pre-eclampsia are neither activated nor primed in vivo, however the release of reactive oxygen species is diminished in normal pregnancy. In comparison, an elevation of reactive oxygen generation in pre-eclampsia may highlight a role for neutrophils in the oxidative stress and pathophysiology of this disease.

Adult↗

Significance of fatty acids in pregnancy-induced immunosuppression.

Pregnancy can exert suppressive effects on chronic inflammatory conditions. We have previously demonstrated a depression in polymorphonuclear leukocyte (PMN) respiratory burst during pregnancy which could explain this amelioration. To elucidate the biochemical mechanism, we have examined PMN phospholipase A2 (PLA2) activity and its relationship to cellular and circulating fatty acids in pregnant women (30 to 34 weeks) and nonpregnant controls. PMN PLA2 activity was determined by arachidonic acid (AA) and leukotriene B4 (LTB4) release, respiratory burst activity was determined by lucigenin-enhanced chemiluminescence, and total serum and PMN fatty acid levels were determined by gas-liquid chromatography. AA release was significantly reduced for pregnancy PMNs in response to N-formyl-met-leu-phe (fMLP) under unprimed and tumor necrosis factor alpha (TNF-alpha)- or interleukin 8-primed conditions. Similarly, LTB4 liberation was significantly reduced in response to fMLP and phorbol myristate acetate in unprimed and TNF-alpha-primed pregnancy PMNs. All major fatty acid classes were altered in the pregnant state. Of these differences in PMNs, oleic acid and alpha-linolenic acid showed a significant increase (13 and 26%, respectively) and stearic acid and AA showed a significant decrease (8 and 30%, respectively). The stearic acid, oleic acid, and AA compositions of all cells analyzed correlated with their corresponding changes in serum fatty acid levels. Crossover serum incubations modified both fatty acid profiles and the PMN respiratory burst accordingly, while individual fatty acid incorporation studies highlighted the importance of polyunsaturated fatty acids for NADPH oxidase efficiency. These findings indicate that the attenuation of PMN function in pregnancy may originate from a reduction in the available pool of cellular fatty acids. Furthermore, this reduction arises as a direct result of a pregnancy-induced shift in circulating fatty acids from polyunsaturated to monounsaturated forms.

Adolescent↗

Role of p38 in the priming of human neutrophils by peritoneal dialysis effluent.

Peritoneal dialysis effluent (PDE) contains a low-molecular-weight substance that is able to prime human neutrophils for the release of arachidonic acid and superoxide anion. Conventional priming agents, such as tumor necrosis factor alpha (TNF-alpha), are known to signal via mitogen-activated protein (MAP) kinases; at least one possible substrate for MAP kinases is cytosolic phospholipase A(2) (cPLA(2)). Phosphorylation of this enzyme results in arachidonic acid release, and this fatty acid is a potent primer and activator of the human neutrophil NADPH oxidase. Because of the striking similarities between the priming of neutrophils with agents such as TNF-alpha and PDE, we have investigated the signalling pathways evoked by PDE and explored the possibility that cPLA(2) is a target for activated MAP kinases. Our results show that PDE treatment of human neutrophils results in the phosphorylation of the p38 kinase rather than the p42 and p44 kinases. Phosphorylation of p38 is transient with maximal activity being observed 1 min after exposure to PDE. We were unable to demonstrate that activation of p38 resulted in phosphorylation of cPLA(2); furthermore, translocation of this enzyme to a membrane-containing fraction was not enhanced in PDE-treated neutrophils. Taken together, these data suggest that, in a manner similar to that of TNF-alpha, PDE primes human neutrophils by the activation of the p38 kinase. However, unlike the cytokine, the activation of this protein does not result in phosphorylation or activation of cPLA(2).

Antibodies↗

The cost of MRI: changes in costs 1989-1996.

A study was conducted to provide an up-to-date analysis of the costs associated with operating an MRI facility in a mainstream National Health Service (NHS) setting. This paper presents an analysis of the costs associated with the acquisition and operation of an MRI scanner installed at the Walsgrave NHS Trust in Coventry in 1988. Costs arising immediately after the introduction of the MRI facility in 1988 are compared with the costs arising towards the end of the scanner's useful life during the 1995/96 financial year. The findings suggest that although the nominal total costs of MRI have increased marginally, from 403,223 Pounds to 434,037 Pounds (an increase of 30,814 Pounds), the increase in total costs is below the rate of inflation, and the nominal average costs of MRI have fallen from 179.20 Pounds to 115.77 Pounds over the period.

Cost-Benefit Analysis↗

Use of opposing reflex stimuli and heart rate variability to examine the effects of lipophilic and hydrophilic beta-blockers on human cardiac vagal control.

Evidence from animal studies suggests that beta-blockers can act within the central nervous system to increase cardiac vagal motoneuron activity. We have attempted to determine whether such an effect is evident in healthy humans, by examining the effects of lipophilic and hydrophilic agents on heart rate variability and cardiac vagal reflexes. A total of 20 healthy volunteers took part in the study. Autonomic studies were performed after 72 h of treatment with placebo, atenolol or metoprolol in a blinded cross-over design. ECG recordings were taken at rest and during mental and orthostatic stress. Heart rate variability was measured in the time and frequency domains. The effects on heart rate of two opposing cardiac vagal reflexes were examined. Trigeminal stimulation causing vagal stimulation, and isometric forearm muscle contraction ('muscle heart reflex') causing vagal inhibition, were performed alone and simultaneously. At rest, during mental stress and during trigeminal stimulation, beta-blocker therapy was associated with significantly increased high-frequency beat-to-beat heart rate variability when compared with placebo. There were no significant differences in effects on heart rate or heart rate variability between atenolol and metoprolol. Analysis of the muscle heart reflex, alone and with simultaneous trigeminal stimulation, showed that the magnitude of the R-R interval response was significantly greater after beta-blocker therapy compared with placebo, but the effects of atenolol and metoprolol were equivalent. beta-Blocker therapy increased cardiac vagal activity, as shown by measures of high-frequency heart rate variability and reflex studies. Lipophilic and hydrophilic beta-blockers appeared to be equally efficacious in increasing the cardiac vagal modulation of heart rate.

Adolescent↗

Sleep states and neonatal pulse oximetry.

AIM: Pulse oximetry is a quick, easy, noninvasive method widely used for monitoring oxygen saturation (SaO2) in the neonatal period. The greatest recognized problem with SaO2 readings measured from the oximeter is artifact, arising from an inability to record accurately during movement. The aim of this study was to determine the proportion of pulse oximeter readings affected by movement artifact during sleep in term and preterm infants. METHOD: Polygraphic recordings were obtained from 11 term and 6 preterm infants at postconceptional ages (SD) of 39.8 (0.8) and 39.3 (1.5) weeks respectively. The polygraphic and computer recordings were divided into epochs of 30 seconds duration and identified as active sleep (AS), quiet sleep (QS), indeterminate sleep (IS), or wakefulness (AW), using electrophysiological parameters and behavioral observations. Movement artifact was identified by visual examination of polygraphic computer recordings using strict criteria. Signal containing artifact was removed from recordings and the percentage of artifact time present per recording calculated. RESULTS: Signal artifact was present in recordings of all infants studied, comprising an average state time of 19% during quiet sleep, 49% of active sleep, 49% of indeterminate sleep and 91% of wakefulness. A significant difference in the proportion of artifact present in recordings of term and preterm was observed only during quiet sleep. CONCLUSION: Movement artifact during pulse oximetry recordings is dependent on behavioral state, and overall affects up to 50% of recorded traces. A reliable and more accurate noninvasive method of recording oxygen saturation is thus needed, for use in both neonatal nurseries and in sleep studies, to aid in accurate clinical decision-making.

Female↗

Development and use of an established cell line of the leafhopper Circulifer tenellus to characterize Spiroplasma citri-vector interactions.

A continuous cell line of embryonic origin from the leafhopper Circulifer tenellus, CT 1, was established using a protocol modified from thrips cell culture. The line was used to develop an in vitro model to examine the mode of entry of the plant pathogenic mollicute Spiroplasma citri into insect host cells. Confluent monolayers were achieved in 5-6 months using a simple medium developed for maintaining established leafhopper cell lines. The newly established CT 1 line, and that of another leafhopper, Nephotettix cincticeps, were exposed to S. citri and examined by electron microscopy. S. citri was found to cytadhere and to be present in apparent invaginations of the host cell membranes of cell lines of both leafhopper species, supporting the hypothesis that this pathogen enters its insect host via endocytosis.

Animals↗

Cytokine degradation by biofilms of Porphyromonas gingivalis.

The aim of this study was to determine whether biofilms of Porphyromonas gingivalis could proteolytically degrade the cytokines interleukin (IL)-1beta, IL-6, or IL-1 receptor antagonist (IL-1ra). Biofilms were grown on membrane filters on the surface of Wilkins-Chalgren blood agar. The biofilms were removed from the plates, and solutions containing 2.5 microg/ml of each cytokine were added. Following incubation for up to 4.0 h, supernatants from the biofilms were subjected to SDS-PAGE. The separated proteins were transferred by Western blotting to PVDF membranes and probed with peroxidase-conjugated antibodies recognizing both the intact cytokines and their degradation products. After 2 h, no intact IL-1beta, IL-6, or IL-1ra were detectable. Cytokine proteolysis also occurred in the presence of horse serum. These results demonstrate that biofilm-grown P. gingivalis can degrade both pro- and anti-inflammatory cytokines and so may be able to perturb cytokine networks in vivo by eliminating cytokines from the local environment.

Biofilms↗

The role of the endogenous anti-inflammatory compound gravidin in pre-eclampsia.

OBJECTIVES: The study's aims were to investigate the levels of gravidin, an endogenous phospholipase A2 inhibitor, in pregnancy and pre-eclampsia and to establish its effects on neutrophil function. STUDY DESIGN: Serum samples were collected from 9 nonpregnant, 15 preeclamptic, and 10 healthy pregnant women and assayed for free gravidin by enzyme-linked immunosorbent assay. Neutrophil phospholipase A2 and respiratory burst activities were determined in the presence of isolated free gravidin by cellular arachidonic acid release and superoxide anion production. RESULTS: Levels of free gravidin were higher in the healthy pregnant (36.1 +/- 5.5 ng/mL) and preeclamptic (17.8 +/- 2.8 ng/mL) groups than in the nonpregnant control group (3.9 +/- 0.5 ng/mL) and were significantly different between pregnancy groups (P <.01, Mann-Whitney U test). Free gravidin caused a concentration dependent decrease in N-formyl-methionyl-leucyl-phenylalanine-stimulated neutrophil arachidonic acid release (inhibitory concentration of 50% 25 nmol/L) and superoxide anion generation (inhibitory concentration of 50% 32 nmol/L). CONCLUSIONS: Circulating levels of free gravidin are reduced in pre-eclampsia compared with normal pregnancy. This may encourage an increase in the respiratory burst of neutrophils in pre-eclampsia and could contribute to the oxidative stress and vascular damage that characterize this disease.

Adult↗

Prognostic factors affecting disease-free survival in patients with Hodgkin's disease stages IA and IIA treated initially with radiotherapy alone in a single centre during 1973 to 1992.

One hundred forty-nine patients with either stage IA (90 patients) or stage IIA (59 patients) Hodgkin's disease (HD) received radiotherapy alone as their first line treatment. Only stage of the nine factors selected for Cox multivariate analysis was of prognostic significance. Patients with grade 1 nodular sclerosing HD did not have a significantly better disease-free survival than those with grade 2 disease.

Adolescent↗

Five-year follow-up of a prospective randomised multi-centre trial of weekly chemotherapy (CAPOMEt) versus cyclical chemotherapy (CHOP-Mtx) in the treatment of aggressive non-Hodgkin's lymphoma. Central Lymphoma Group.

BACKGROUND: Weekly alternating regimen known as CAPOMEt is compared to standard cyclical chemotherapy (CHOP-Mtx) in aggressive non-Hodgkin's lymphoma (NHL). PATIENTS AND METHODS: Three hundred and eighty-one patients with aggressive NHL were randomised to receive either cyclophosphamide, doxorubicin, vincristine, prednisone and methotrexate (CHOP-Mtx) on a cyclical basis or a weekly regimen incorporating the same drugs with the addition of etoposide (CAPOMEt). RESULTS: After pathological review, 281 patients were deemed eligible. At the census date of 31 March 1994, 158 patients were alive with a median follow up of 5.9 years (minimum 3.0 years). Analysis of all patients and eligible patients showed no significant treatment differences in the rates of complete remission (CR), failure free survival (FFS) or overall survival (OS) between the two arms. The actuarial median OS was 24 months for CAPOMEt compared with 31 months for CHOP-Mtx, with five-year actuarial survival rates of 37% and 43%, respectively. Myelosuppression was significantly more severe with CHOP-Mtx and neurotoxicity was much more common with CAPOMEt. CONCLUSION: Weekly CAPOMEt is equally effective as standard cyclical CHOP-Mtx treatment in aggressive NHL.

Adolescent↗

Effects of r-metHuG-CSF on polymorphonuclear leucocyte kinetics and function in patients on continuous ambulatory peritoneal dialysis.

End-stage renal failure (ESRF) patients undergoing continuous ambulatory peritoneal dialysis (CAPD) are immunocompromised and exhibit abnormal circulating polymorphonuclear leucocyte (PMN) function, including reduced phagocytosis and intracellular killing. Six uraemic patients on CAPD were each given 300 microg granulocyte colony stimulating factor (G-CSF) every day for 5 d and PMN function tests were performed daily. By day 5 of the study CD11b expression was significantly decreased in response to N-formylmethionylleucylphenylalanine (fMLP) and opsonized Staphylococcus epidermidis stimulation, and expression of L-selectin (CD62L) was significantly decreased in response to opsonized Staphylococcus epidermidis stimulation. Further, superoxide anion production and Fc gammaRI (CD64) expression were found to be significantly increased and Fc gammaRII (CD16) expression was lowered. Circulating white cell and PMN counts were significantly elevated in response to treatment. Administration of G-CSF did not appear to have corrected the abnormalities in phagocytosis and intracellular killing. This study suggests that G-CSF does no harm to ESRF patients and influences uraemic PMN function in a manner that is comparable to its effects on PMN in non-uraemic subjects.

Adult↗

A comparison of medical and surgical termination of pregnancy: choice, emotional impact and satisfaction with care.

OBJECTIVE: To investigate whether women having medical or surgical terminations of pregnancy differ in their emotional distress before or after the procedure. To evaluate whether choice of procedure occurs, the factors influencing type of procedure and the effect of choice on emotional responses and satisfaction with care. DESIGN: A prospective comparative study. SETTING: A termination of pregnancy unit in a University Teaching Hospital. PARTICIPANTS: Two hundred and seventy-five women attending for medical or surgical first trimester termination of pregnancy. METHODS: Interviews concerning choice and measures of emotional status were completed prior to terminations. Four weeks after termination emotional functioning was reassessed together with satisfaction with care. RESULTS: Women having a surgical termination waited longer for the procedure and were at more advanced gestation than those having the medical termination. There were no differences in emotional responses related to type of procedure or gestation. One-quarter remained highly anxious at four weeks. Medical and surgical groups did not differ in emotional status prior to termination. Those having the medical procedure rated it as marginally more stressful and experienced more post-termination physical problems and disruption to life. Seeing the fetus was associated with more intrusive events (nightmares, flashbacks, unwanted thoughts related to the experience). One-quarter of the medical and 67% of the surgical group reported having no choice in type of procedure. Only 53% of the medical group would choose the same procedure again compared with 77% of the surgical group. CONCLUSIONS: Termination method did not influence emotional adjustment. Many women were not offered genuine choice of procedure. Having choice was considered very important but was unrelated to emotional distress or satisfaction with care.

Abortifacient Agents↗

Role of arachidonic acid and its metabolites in the priming of NADPH oxidase in human polymorphonuclear leukocytes by peritoneal dialysis effluent.

Peritoneal dialysis effluent (PDE) contains a low-molecular-weight solute that will activate and prime the NADPH oxidase of human neutrophils via a phospholipase A2 (PLA2)-dependent mechanism. Since the products of PLA2 are known to activate and prime the oxidase we have investigated their role in the dialysis effluent-mediated activation and priming of human neutrophils. NADPH oxidase activity of PDE-primed and -unprimed neutrophils was measured by lucigenin-enhanced chemiluminescence in the presence of known inhibitors of the arachidonic acid cascade. Incubation of neutrophils with the nonselective PLA2 inhibitor quinacrine (0 to 100 microM) reduced oxidase activity in both primed and unprimed cells. Furthermore, primed cells were more sensitive to the action of quinacrine than were unprimed cells. We were unable to determine the relative roles of secretory PLA2 (sPLA2) and cytosolic PLA2 (cPLA2) since the selective sPLA2 inhibitor scalaradial (0 to 100 microM) inhibited oxidase activity in both groups of cells by similar degrees, while the specific cPLA2 inhibitor AACO-CF3 (0 to 50 microM) failed to affect activity in either group. Inhibition of platelet-activating factor (PAF), cycloxygenase, and 5-lipoxygenase-activating protein by hexanolamino-PAF (0 to 25 microM), flurbiprofen (0 to 25 microM), and MK886 (0 to 5 microM), respectively, had no effect upon oxidase activity. However, the direct inhibition of 5-lipoxygenase by caffeic acid or lipoxin A4 resulted in a similar concentration-dependent attenuation of oxidase activity in both primed and unprimed cells. Leukotriene B4 (LTB4) release from primed neutrophils was comparable to that from unprimed cells with the exception of phorbol myristate acetate-stimulated cells, which released fivefold more LTB4 than control. Taken together, these results suggest that it is arachidonic acid per se, and not its metabolites, that is important in priming of the neutrophil NADPH oxidase by dialysis effluent.

Anti-Inflammatory Agents↗

Systematic review of the school entry medical examination.

AIMS: To summarise and critically evaluate research conducted in the UK between 1962 and 1996, on the effectiveness and efficiency of the school entry medical (SEM) examination. METHODS: An electronic search of a large number of databases, in conjunction with a search of reference lists, and sources in the grey literature produced a total of 64 studies. RESULTS: Only one overview and 16 primary studies met the review's broad inclusion criteria. The results showed significant differences in the identification and referral of new and ongoing problems not only between the routine and selective SEM but also within the two types of SEM examination. There were also large differences in the numbers of children selected for SEM examination. No study included in the review defined either the methods or the criteria used to identify children as screen positive. No study provided follow up of children after referral to estimate the positive predictive value or yield of the screening, or follow up of the whole cohort to identify false negative cases. CONCLUSION: Data on the effectiveness and efficiency of both the routine and selective SEM examination in accurately identifying children with new or ongoing health problems are not available at the present time. The studies reviewed here demonstrate the fragility of the evidence on which the school entry medical is based, and call into question the ethical basis of this programme.

Child, Preschool↗

Angiotensin II modulates cardiovascular autonomic control in the absence of baroreflex loading.

OBJECTIVE: To investigate the effects of angiotensin II in the absence of baroreflex activation. DESIGN: Ten healthy male volunteers were studied in a single blind, randomised, crossover study of heart rate variability during intravenous angiotensin II infusion (5-20 ng/kg/min) compared with a control pressor infusion of phenylephrine (0.7-2.8 micrograms/kg/min). Each infusion was titrated to increase mean blood pressure by 20 mm Hg; sodium nitroprusside was then infused simultaneously to restore blood pressure to baseline values. RESULTS: During concomitant angiotensin II (AII) and sodium nitroprusside (SNP) infusion, the mean (SD) RR interval (864 (117) ms) was significantly shorter than during phenylephrine (PE) and sodium nitroprusside infusion (1057 (163) ms), and was significantly shorter than at baseline (999 (164) ms), despite comparable levels of blood pressure. Values of high frequency heart rate variability measured in the time and frequency domains were significantly lower during AII/SNP infusion than during PE/SNP: percentage of successive RR interval differences exceeding 50 ms, 30(16)% v 57(21)%; root mean square of successive RR interval differences, 63 (39) v 90 (40) ms; high frequency power 0.48 (0.19) v 0.66 (0.26) nu. CONCLUSIONS: When the pressor response is controlled by sodium nitroprusside, angiotensin II infusion is associated with tachycardia. Analysis of heart rate variability suggests that this reflects inhibition of cardiac vagal activity.

Adolescent↗