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1.
BMC Public Health ; 24(1): 800, 2024 Mar 14.
Artigo em Inglês | MEDLINE | ID: mdl-38481177

RESUMO

BACKGROUND: Local authorities in England have an important role in shaping healthy local environments contributing to childhood obesity. This study examined changes in diet and physical activity in primary school children following a three-year, complex, community-based intervention in Golborne ward, the second most deprived ward in London. METHODS: The Go-Golborne intervention aimed to shape the local environment across multiple settings with the engagement of a large number of local government and community stakeholders in a joint approach. Activities focused on six co-created themes to make changes to local environments and reduce sugary snacks and beverage consumption, increase fruit and vegetable intake, promote healthy snacks, increase active play and travel, and reduce screen time. We analysed changes in self-reported diet and physical activity, collected annually between 2016 and 2019, from 1,650 children aged 6-11 years through six local schools, who all received the intervention. We used multilevel, linear and logistic random-slope regression models adjusted for time on study, baseline age, gender, ethnicity, deprivation quintile, school, and baseline weight status. RESULTS: After three years of follow-up, there were reductions in sugar-sweetened beverage consumption (adjusted beta -0·43 occasions/day, 95% CI -0·55 to -0·32), fruit and vegetable consumption (adjusted beta -0.22 portions, 95% CI -0.44 to 0.001) and car travel to and from school (adjusted OR 0·19, 95% CI 0·06 to 0·66), while screen time increased (high versus moderate/low: OR 2·30, 95% CI 1·36 to 3·90). For other behavioural outcomes, there was no statistically significant evidence of changes. CONCLUSION: Local authorities have substantial powers to make positive changes to the obesogenic environment but programmes remain under-evaluated. Results from the ambitious Go-Golborne intervention demonstrated mixed results in health behaviours following programme implementation. These results underline the importance of a coordinated and comprehensive policy response to support changes in wider environmental and social conditions as well as appropriate and holistic evaluations of initiatives to inform local actions on obesogenic environments.


Assuntos
Obesidade Pediátrica , Criança , Humanos , Dieta , Exercício Físico , Londres/epidemiologia , Obesidade Pediátrica/epidemiologia , Obesidade Pediátrica/prevenção & controle , Projetos Piloto , Masculino , Feminino
2.
BMC Public Health ; 23(1): 2547, 2023 12 20.
Artigo em Inglês | MEDLINE | ID: mdl-38124041

RESUMO

BACKGROUND: Air pollution remains a significant public health risk, particularly in urban areas. Effective communication strategies remain integral to overall protection by encouraging the adoption of personal air pollution exposure reduction behaviours. This study aims to explore how community health professionals can be empowered to communicate air pollution information and advice to the wider community, to encourage the uptake of desired behaviours in the population. METHODS: The study adopted a qualitative methodology, where four homogenous Focus Group Discussions (FGDs) were held with a range of community health professionals, including Health Care Professionals, Community Health and Wellbeing Workers (CHWWs) and Social Prescribing Link Workers (SPLW). A classical content analysis was conducted with the Structural Empowerment Theory (SET) and Psychological Empowerment Theory (PET) as guiding concepts. RESULTS: Five key themes were identified: from a structural empowerment perspective: [1] resources and support, [2] knowledge. From a psychological empowerment perspective: [3] confidence as advisor, [4] responsibility as advisor, and [5] residents' receptiveness to advice. It was concluded that advice should be risk stratified, clear, easy to follow and provide alternatives. CONCLUSION: This study identified ways for community health professionals to be empowered by local councils or other organisations in providing advice on air pollution, through the provision of essential structural support and opportunities to enhance their knowledge and confidence in the subject. Implementing recommendations from this study would not only empower community health professionals to advise on air pollution to the wider community but also increase adherence to health advice.


Assuntos
Poluição do Ar , Saúde Pública , Humanos , Poluição do Ar/efeitos adversos , Poluição do Ar/prevenção & controle , Poluição do Ar/análise , Pessoal de Saúde , Comunicação , Poder Psicológico
3.
BMC Health Serv Res ; 23(1): 1092, 2023 Oct 12.
Artigo em Inglês | MEDLINE | ID: mdl-37821938

RESUMO

BACKGROUND: Delays in preventative service uptake are increasing in the UK. Universal, comprehensive monthly outreach by Community Health and Wellbeing Workers (CHW), who are integrated at the GP practice and local authority, offer a promising alternative to general public health campaigns as it personalises health promotion and prevention of disease holistically at the household level. We sought to test the ability of this model, which is based on the Brazilian Family Health Strategy, to increase prevention uptake in the UK. METHODS: Analysis of primary care patient records for 662 households that were allocated to five CHWWs from July 2021. Primary outcome was the Composite Referral Completion Indicator (CRCI), a measure of how many health promotion activities were received by members of a household relative to the ones that they were eligible for during the period July 2021-April 2022. The CRCI was compared between the intervention group (those who had received at least one visit) and the control group (allocated households that were yet to receive a visit). A secondary outcome was the number of GP visits in the intervention and control groups during the study period and compared to a year prior. RESULTS: Intervention and control groups were largely comparable in terms of household occupancy and service eligibilities. A total of 2251 patients in 662 corresponding households were allocated to 5 CHWs and 160 households had received at least one visit during the intervention period. The remaining households were included in the control group. Overall service uptake was 40% higher in the intervention group compared to control group (CRCI: 0.21 ± 0.15 and 0.15 ± 0.19 respectively). Likelihood of immunisation uptake specifically was 47% higher and cancer screening and NHS Health Checks was 82% higher. The average number of GP consultations per household decreased by 7.4% in the intervention group over the first 10 months of the pilot compared to the 10 months preceding its start, compared with a 0.6% decrease in the control group. CONCLUSIONS: Despite the short study period these are promising findings in this deprived, traditionally hard to reach community and demonstrates potential for the Brazilian community health worker model to be impactful in the UK. Further analysis is needed to examine if this approach can reduce health inequalities and increase cost effectiveness of health promotion approaches.


Assuntos
Detecção Precoce de Câncer , Serviços Preventivos de Saúde , Saúde Pública , Medicina Estatal , Vacinação , Humanos , Brasil , Agentes Comunitários de Saúde/estatística & dados numéricos , Detecção Precoce de Câncer/estatística & dados numéricos , Neoplasias/diagnóstico , Neoplasias/epidemiologia , Saúde Pública/estatística & dados numéricos , Medicina Estatal/estatística & dados numéricos , Reino Unido/epidemiologia , Relações Comunidade-Instituição , Serviços Preventivos de Saúde/organização & administração
4.
J Public Health (Oxf) ; 45(4): 878-887, 2023 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-37608490

RESUMO

BACKGROUND: Commercial advertising and sponsorship drive the consumption of harmful commodities. Local authorities (LAs) have considerable powers to reduce such exposures. This study aimed to characterize local commercial policies across all English LAs. METHODS: We conducted a census of all English LAs (n = 333) to identify local commercial policies concerning advertising and sponsorship of tobacco, alcohol, less healthy foods and gambling, through online searches and Freedom of Information requests. We explored policy presence, commodity frequency and type, and associations with LA characteristics (region, urban/rural and deprivation). RESULTS: Only a third (106) of LAs in England had a relevant policy (32%). These included restrictions on tobacco (91%), gambling (79%), alcohol (74%) and/or less healthy foods (24%). Policy prevalence was lowest in the East of England (22%), North East (25%) and North West (27%), higher in urban areas (36%) than rural areas (28%) and lower in the least (27%) compared with the most (38%) deprived areas. Definitions in policies varied, particularly for alcohol and less healthy foods. CONCLUSIONS: English LAs currently underutilize their levers to reduce the negative impacts of harmful commodity industry marketing, particularly concerning less healthy foods. Standardized guidance, including clarity on definitions and application, could inform local policy development.


Assuntos
Publicidade , Governo Local , Humanos , Indústrias , Marketing , Políticas , Determinantes Sociais da Saúde
5.
BMJ Glob Health ; 8(1)2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36690378

RESUMO

INTRODUCTION: Industries that produce and market potentially harmful commodities or services (eg, tobacco, alcohol, gambling, less healthy foods and beverages) are a major influence on the drivers of behavioural risk factors for non-communicable diseases. The nature and impact of interactions between public bodies and 'harmful commodity industries' (HCIs) has been widely recognised and discussed at national and international levels, but to date little is known about such interactions at local or regional government levels. This study aimed to identify and characterise actual and potential interactions and proposes a typology of interactions between HCIs and English local authorities (LAs). METHODS: Five electronic databases covering international literature (PubMed, EBSCO, OVID, Scopus and Web of Science) were searched up to June 2021. We also performed online searches for publicly available, web-based grey literature and documented examples of interactions in an English LA context. We conducted a critical interpretive synthesis of the published and grey literature to integrate and conceptualise the data in the context of English LAs. RESULTS: We included 47 published papers to provide the frame for the typology, which was refined and contextualised for English LAs through the available grey literature. Three categories were developed, describing the medium through which interactions occur: (1) direct involvement with LAs, (2) involvement through intermediaries and (3) involvement through the local knowledge space. Within these, we grouped interactions into 10 themes defining their nature and identified illustrative examples. CONCLUSION: Our typology identifies complex inter-relationships and characterises interactions between HCIs and LAs, with illustrative examples from English LAs. Drawn from well-established theories and frameworks in combination with contextual information on English LAs, this typology explores the LA perspective and could help local decision-makers to maximise population health while minimising negative impacts of HCIs. PROSPERO REGISTRATION NUMBER: CRD42021257311.


Assuntos
Governo Local , Humanos , Fatores de Risco , Inglaterra
6.
Age Ageing ; 51(3)2022 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-35231093

RESUMO

BACKGROUND: An ageing population and limited resources have put strain on state provision of adult social care (ASC) in England. With social care needs predicted to double over the next 20 years, there is a need for new approaches to inform service planning and development, including through predictive models of demand. OBJECTIVE: Describe risk factors for long-term ASC in two inner London boroughs and develop a risk prediction model for long-term ASC. METHODS: Pseudonymised person-level data from an integrated care dataset were analysed. We used multivariable logistic regression to model associations of demographic factors, and baseline aspects of health status and health service use, with accessing long-term ASC over 12 months. RESULTS: The cohort comprised 13,394 residents, aged ≥75 years with no prior history of ASC at baseline. Of these, 1.7% became ASC clients over 12 months. Residents were more likely to access ASC if they were older or living in areas with high socioeconomic deprivation. Those with preexisting mental health or neurological conditions, or more intense prior health service use during the baseline period, were also more likely to access ASC. A prognostic model derived from risk factors had limited predictive power. CONCLUSIONS: Our findings reinforce evidence on known risk factors for residents aged 75 or over, yet even with linked routinely collected health and social care data, it was not possible to make accurate predictions of long-term ASC use for individuals. We propose that a paradigm shift towards more relational, personalised approaches, is needed.


Assuntos
Assistência de Longa Duração , Saúde Mental , Estudos de Coortes , Humanos , Estudos Retrospectivos , Apoio Social
10.
Photosynth Res ; 135(1-3): 141-142, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-28929465

RESUMO

In Fig. 1a in the original article, the amino acid side chains were incorrectly labeled in the structure representation of the orange carotenoid protein (OCP). The corrected figure is printed in this erratum.

11.
Photosynth Res ; 135(1-3): 125-139, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28236074

RESUMO

Photoprotection in cyanobacteria relies on the interplay between the orange carotenoid protein (OCP) and the fluorescence recovery protein (FRP) in a process termed non-photochemical quenching, NPQ. Illumination with blue-green light converts OCP from the basic orange state (OCPO) into the red-shifted, active state (OCPR) that quenches phycobilisome (PBs) fluorescence to avoid excessive energy flow to the photosynthetic reaction centers. Upon binding of FRP, OCPR is converted to OCPO and dissociates from PBs; however, the mode and site of OCPR/FRP interactions remain elusive. Recently, we have introduced the purple OCPW288A mutant as a competent model for the signaling state OCPR (Sluchanko et al., Biochim Biophys Acta 1858:1-11, 2017). Here, we have utilized fluorescence labeling of OCP at its native cysteine residues to generate fluorescent OCP proteins for fluorescence correlation spectroscopy (FCS). Our results show that OCPW288A has a 1.6(±0.4)-fold larger hydrodynamic radius than OCPO, supporting the hypothesis of domain separation upon OCP photoactivation. Whereas the addition of FRP did not change the diffusion behavior of OCPO, a substantial compaction of the OCPW288A mutant and of the OCP apoprotein was observed. These results show that sufficiently stable complexes between FRP and OCPW288A or the OCP apoprotein are formed to be detected by FCS. 1:1 complex formation with a micromolar apparent dissociation constant between OCP apoprotein and FRP was confirmed by size-exclusion chromatography. Beyond the established OCP/FRP interaction underlying NPQ cessation, the OCP apoprotein/FRP interaction suggests a more general role of FRP as a scaffold protein for OCP maturation.


Assuntos
Apoproteínas/metabolismo , Proteínas de Bactérias/metabolismo , Transdução de Sinais , Sequência de Aminoácidos , Apoproteínas/química , Proteínas de Bactérias/química , Varredura Diferencial de Calorimetria , Cromatografia em Gel , Cisteína/metabolismo , Difusão , Hidrodinâmica , Espectrometria de Massas , Reprodutibilidade dos Testes , Espectrometria de Fluorescência , Coloração e Rotulagem , Compostos de Sulfidrila/metabolismo
13.
Biophys J ; 113(10): 2249-2260, 2017 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-28988699

RESUMO

The Na+,K+-ATPase is a plasma membrane ion transporter of high physiological importance for ion homeostasis and cellular excitability in electrically active tissues. Mutations in the genes coding for Na+,K+-ATPase α-subunit isoforms lead to severe human pathologies including Familial Hemiplegic Migraine type 2, Alternating Hemiplegia of Childhood, Rapid-onset Dystonia Parkinsonism, or epilepsy. Many of the reported mutations lead to change- or loss-of-function effects, whereas others do not alter the functional properties, but lead to, e.g., reduced protein stability, reduced protein expression, or defective plasma membrane targeting. Na+,K+-ATPase frequently assembles with other membrane transporters or cellular matrix proteins in specialized plasma membrane microdomains, but the effects of these interactions on targeting or protein mobility are elusive so far. Mutation of established interaction motifs of the Na+,K+-ATPase with ankyrin B and caveolin-1 are expected to result in changes in plasma membrane targeting, changes of the localization pattern, and of the diffusion behavior of the enzyme. We studied the consequences of mutations in these binding sites by monitoring diffusion of eGFP-labeled Na+,K+-ATPase constructs in the plasma membrane of HEK293T cells by fluorescence correlation spectroscopy as well as fluorescence recovery after photobleaching or photoswitching, and observed significant differences compared to the wild-type enzyme, with synergistic effects for combinations of interaction site mutations. These measurements expand the possibilities to study the consequences of Na+,K+-ATPase mutations and provide information about the interaction of Na+,K+-ATPase α-isoforms with cellular matrix proteins, the cytoskeleton, or other membrane protein complexes.


Assuntos
Anquirinas/química , Anquirinas/metabolismo , Caveolina 1/química , Caveolina 1/metabolismo , Membrana Celular/metabolismo , Mutação , ATPase Trocadora de Sódio-Potássio/metabolismo , Sequência de Aminoácidos , Animais , Transporte Biológico/genética , Difusão , Células HEK293 , Humanos , Modelos Moleculares , Oócitos/metabolismo , Ligação Proteica/genética , Domínios Proteicos , Rubídio/metabolismo , ATPase Trocadora de Sódio-Potássio/química , ATPase Trocadora de Sódio-Potássio/genética , Xenopus laevis/metabolismo
14.
Front Physiol ; 7: 239, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27445835

RESUMO

Mutations in four genes have been identified in familial hemiplegic migraine (FHM), from which CACNA1A (FHM type 1) and SCN1A (FHM type 3) code for neuronal voltage-gated calcium or sodium channels, respectively, while ATP1A2 (FHM type 2) encodes the α2 isoform of the Na(+),K(+)-ATPase's catalytic subunit, thus classifying FHM primarily as an ion channel/ion transporter pathology. FHM type 4 is attributed to mutations in the PRRT2 gene, which encodes a proline-rich transmembrane protein of as yet unknown function. The Na(+),K(+)-ATPase maintains the physiological gradients for Na(+) and K(+) ions and is, therefore, critical for the activity of ion channels and transporters involved neuronal excitability, neurotransmitter uptake or Ca(2+) signaling. Strikingly diverse functional abnormalities have been identified for disease-linked ATP1A2 mutations which frequently lead to changes in the enzyme's voltage-dependent properties, kinetics, or apparent cation affinities, but some mutations are truly deleterious for enzyme function and thus cause full haploinsufficiency. Here, we summarize structural and functional data about the Na(+),K(+)-ATPase available to date and an overview is provided about the particular properties of the α2 isoform that explain its physiological relevance in electrically excitable tissues. In addition, current concepts about the neurobiology of migraine, the correlations between primary brain dysfunction and mechanisms of headache pain generation are described, together with insights gained recently from modeling approaches in computational neuroscience. Then, a survey is given about ATP1A2 mutations implicated in migraine cases as documented in the literature with focus on mutations that were described to completely destroy enzyme function, or lead to misfolded or mistargeted protein in particular model cell lines. We also discuss whether or not there are correlations between these most severe mutational effects and clinical phenotypes. Finally, perspectives for future research on the implications of Na(+),K(+)-ATPase mutations in human pathologies are presented.

15.
Eur J Emerg Med ; 23(2): 130-6, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25380318

RESUMO

OBJECTIVE: We aimed to compare the performance of a widely used physiological score [Modified Early Warning Score (MEWS)] and a novel metabolic score (derived from a blood gas) in predicting outcome in emergency department patients. DESIGN, SETTING AND PARTICIPANTS: We carried out a prospective observational study using a convenience sample of 200 patients presenting to the resuscitation area of an inner-city teaching hospital over 4 months. MAIN OUTCOME MEASURES: We looked primarily at whether either score predicted new organ failure at 48 h. Our secondary outcome measures were escalation of care and mortality at 48 h. RESULTS: In univariate analysis, MEWS and the metabolic score predicted 48-h organ failure [odds ratio (OR) 1.19, 95% confidence interval (CI) 1.04-1.35, P=0.009, and OR 1.34, 95% CI 1.015-1.56, P<0.001, respectively]. Both MEWS and the metabolic score predicted 48-h death (OR 1.32, 95% CI 1.02-1.71, P=0.03, and OR 1.56, 95% CI 1.18-2.06, P=0.002, respectively) in univariate analysis. Neither predicted 48-h escalation of care. The metabolic score remained statistically significant at predicting organ failure or death after controlling for MEWS parameters (OR 1.35, 95% CI 1.13-1.62, P=0.001, and OR 1.74, 95% CI 1.13-2.69, P=0.01, respectively). In contrast, MEWS was no longer associated with these outcomes; however, our study has small participant numbers. CONCLUSION: This pilot data suggest that a blood gas-derived metabolic score on emergency department arrival may be superior to MEWS at predicting organ failure and death at 48 h.


Assuntos
Serviço Hospitalar de Emergência , Hospitalização/estatística & dados numéricos , Testes Imediatos , Índice de Gravidade de Doença , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Serviço Hospitalar de Emergência/estatística & dados numéricos , Feminino , Mortalidade Hospitalar , Humanos , Masculino , Pessoa de Meia-Idade , Insuficiência de Múltiplos Órgãos/epidemiologia , Testes Imediatos/estatística & dados numéricos , Valor Preditivo dos Testes , Ressuscitação/estatística & dados numéricos , Resultado do Tratamento , Adulto Jovem
16.
Eur Heart J Qual Care Clin Outcomes ; 1(1): 37-43, 2015 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-29474566

RESUMO

AIMS: Silent myocardial ischaemia occurs commonly in diabetes. Whether altered perception of ischaemia also predisposes to atypical presentations with under-diagnosis of coronary disease is not known. To determine whether (i) patients with diabetes diagnosed with angina are more likely to report atypical symptoms compared with patients without diabetes, and (ii) atypical symptoms in patients with diabetes cause angina to go unrecognized, increasing the risk of coronary events. METHODS AND RESULTS: Prospective, multicentre cohort study of 8662 ambulatory patients with suspected angina, of whom 906 had diabetes. We recorded detailed chest pain descriptors and fatal and non-fatal coronary events over a median of 3.08 years of follow-up. Proportionately more patients with than without diabetes received a diagnosis of angina (42.7 vs. 25.1%). Among patients with diabetes diagnosed with angina, a greater proportion had atypical chest pain compared with patients without diabetes (21.0 vs. 11.3%), but the hazard of fatal and non-fatal coronary events was similar. However, among patients diagnosed with non-cardiac chest pain, those with diabetes-most of whom had atypical symptoms-remained at greater risk of coronary events [2.29 (95% CI 1.54, 3.41)] and all-cause mortality [1.67 (95% confidence interval, CI 1.04, 2.69)] compared with non-diabetic patients. CONCLUSION: Patients with diabetes and atypical symptoms are nearly twice as likely to be diagnosed with angina compared with non-diabetic patients. Those diagnosed with non-cardiac pain are at increased risk of coronary events. Our study emphasizes the need for more intensive investigation of diabetic patients with chest pain, particularly those presenting with atypical symptoms.

17.
PLoS One ; 9(6): e99925, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24937680

RESUMO

BACKGROUND: International guidelines support an early invasive management strategy (including early coronary angiography and revascularisation) for non-ST-elevation acute coronary syndrome (NSTE-ACS) in patients with renal impairment. However, evidence from outside the UK suggests that this approach is underutilised. We aimed to describe practice within the NHS, and to determine whether the severity of renal dysfunction influenced the provision of angiography and modified the association between early revascularisation and survival. METHODS: We performed a cohort study, using multivariable logistic regression and propensity score analyses, of data from the Myocardial Ischaemia National Audit Project for patients presenting with NSTE-ACS to English or Welsh hospitals between 2008 and 2010. FINDINGS: Of 35 881 patients diagnosed with NSTE-ACS, eGFR of <60 ml/minute/1.73 m(2) was present in 15 680 (43.7%). There was a stepwise decline in the odds of undergoing inpatient angiography with worsening renal dysfunction. Compared with an eGFR>90 ml/minute/1.73 m(2), patients with an eGFR between 45-59 ml/minute/1.73 m(2) were 33% less likely to undergo angiography (adjusted OR 0.67, 95% CI 0.55-0.81); those with an eGFR<30/minute/1.73 m(2) had a 64% reduction in odds of undergoing angiography (adjusted OR 0.36, 95%CI 0.29-0.43). Of 16 646 patients who had inpatient coronary angiography, 58.5% underwent inpatient revascularisation. After adjusting for co-variables, inpatient revascularisation was associated with approximately a 30% reduction in death within 1 year compared with those managed medically after coronary angiography (adjusted OR 0.66, 95%CI 0.57-0.77), with no evidence of modification by renal function (p(interaction) = 0.744). INTERPRETATION: Early revascularisation may offer a similar survival benefit in patients with and without renal dysfunction, yet renal impairment is an important determinant of the provision of coronary angiography following NSTE-ACS. A randomised controlled trial is needed to evaluate the efficacy of an early invasive approach in patients with severe renal dysfunction to ensure that all patients who may benefit are offered this treatment option.


Assuntos
Síndrome Coronariana Aguda/mortalidade , Isquemia Miocárdica/mortalidade , Insuficiência Renal/mortalidade , Síndrome Coronariana Aguda/diagnóstico por imagem , Síndrome Coronariana Aguda/fisiopatologia , Síndrome Coronariana Aguda/cirurgia , Idoso , Idoso de 80 Anos ou mais , Estudos de Coortes , Angiografia Coronária , Feminino , Taxa de Filtração Glomerular , Humanos , Pacientes Internados , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Isquemia Miocárdica/fisiopatologia , Isquemia Miocárdica/cirurgia , Intervenção Coronária Percutânea , Pontuação de Propensão , Insuficiência Renal/fisiopatologia , Resultado do Tratamento
18.
Biochim Biophys Acta ; 1837(9): 1581-93, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24742974

RESUMO

The determination of pH in the cell cytoplasm or in intracellular organelles is of high relevance in cell biology. Also in plant cells, organelle-specific pH monitoring with high spatial precision is an important issue, since e.g. ΔpH across thylakoid membranes is the driving force for ATP synthesis critically regulating photoprotective mechanisms like non-photochemical quenching (NPQ) of chlorophyll (Chl) fluorescence or the xanthophyll cycle. In animal cells, pH determination can serve to monitor proton permeation across membranes and, therefore, to assay the efficiency of drugs against proton-selective transporters or ion channels. In this work, we demonstrate the applicability of the pH-sensitive GFP derivative (eGFP-pHsens, originally termed deGFP4 by Hanson et al. [1]) for pH measurements using fluorescence lifetime imaging microscopy (FLIM) with excellent precision. eGFP-pHsens was either expressed in the cytoplasm or targeted to the mitochondria of Chinese hamster ovary (CHO-K1) cells and applied here for monitoring activity of the M2 proton channel from influenza A virus. It is shown that the M2 protein confers high proton permeability of the plasma membrane upon expression in CHO-K1 cells resulting in rapid and strong changes of the intracellular pH upon pH changes of the extracellular medium. These pH changes are abolished in the presence of amantadine, a specific blocker of the M2 proton channel. These results were obtained using a novel multi-parameter FLIM setup that permits the simultaneous imaging of the fluorescence amplitude ratios and lifetimes of eGFP-pHsens enabling the quick and accurate pH determination with spatial resolution of 500 nm in two color channels with time resolution of below 100 ps. With FLIM, we also demonstrate the simultaneous determination of pH in the cytoplasm and mitochondria showing that the pH in the mitochondrial matrix is slightly higher (around 7.8) than that in the cytoplasm (about 7.0). The results obtained for CHO-K1 cells without M2 channels in comparison to M2-expressing cells show that the pH dynamics is determined by the specific H⁺ permeability of the membrane, the buffering of protons in the internal cell lumen and/or an outwardly directed proton pump activity that stabilizes the interior pH at a higher level than the external acidic pH. This article is part of a special issue entitled: photosynthesis research for sustainability: keys to produce clean energy.


Assuntos
Proteínas de Fluorescência Verde/metabolismo , Microscopia de Fluorescência/métodos , Sequência de Aminoácidos , Animais , Células CHO , Cricetinae , Cricetulus , Concentração de Íons de Hidrogênio , Dados de Sequência Molecular , Fotossíntese
19.
BMJ Open ; 2(3)2012.
Artigo em Inglês | MEDLINE | ID: mdl-22700834

RESUMO

OBJECTIVES: To determine whether access to rapid access chest pain clinics of people with recent onset symptoms is equitable by age, socioeconomic status, ethnicity and gender, according to need. DESIGN: Retrospective cohort study with ecological analysis. SETTING: Patients referred from primary care to five rapid access chest pain clinics in secondary care, across England. PARTICIPANTS: Of 8647 patients aged ≥35 years referred to chest pain clinics with new-onset stable chest pain but no known cardiac history, 7570 with documented census ward codes, age, gender and ethnicity comprised the study group. Patients excluded were those with missing date of birth, gender or ethnicity (n=782) and those with missing census ward codes (n=295). OUTCOME MEASURES: Effects of age, gender, ethnicity and socioeconomic status on clinic attendance were calculated as attendance rate ratios, with number of attendances as the outcome and resident population-years as the exposure in each stratum, using Poisson regression. Attendance rate ratios were then compared with coronary heart disease (CHD) mortality ratios to determine whether attendance was equitable according to need. RESULTS: Adjusted attendance rate ratios for patients aged >65 years were similar to younger patients (1.1, 95% CI 1.05 to 1.16), despite population CHD mortality rate ratios nearly 15 times higher in the older age group. Women had lower attendance rate ratios (0.81, 95% CI 0.77 to 0.84) and also lower population CHD mortality rate ratios compared with men. South Asians had higher attendance rates (1.67, 95% CI 1.57 to 1.77) compared with whites and had a higher standardised CHD mortality ratio of 1.46 (95% CI 1.41 to 1.51). Although univariable analysis showed that the most deprived patients (quintile 5) had an attendance rate twice that of less deprived quintiles, the adjusted analysis showed their attendance to be 13% lower (0.87, 95% CI 0.81 to 0.94) despite a higher population CHD mortality rate. CONCLUSION: There is evidence of underutilisation of chest pain clinics by older people and those from lower socioeconomic status. More robust and patient focused administrative pathways need to be developed to detect inequity, correction of which has the potential to substantially reduce coronary mortality.

20.
J Vasc Access ; 11(2): 132-7, 2010.
Artigo em Inglês | MEDLINE | ID: mdl-20155716

RESUMO

OBJECTIVE: Although European Best Practice Guidelines on vascular access recommend universal pre-operative duplex scan in patients receiving brachio-cephalic (BC) arteriovenous fistulae (AVF), this is not widespread practice. Furthermore, cadaveric and angiographic studies suggest that variation in upper limb arterial anatomy is common. Our aim was to investigate the prevalence of high brachial artery bifurcation (HB) and its impact on BC AVF patency. METHODS: A retrospective analysis of consecutive autologous BC AVF created over an 18-month period (January 2008 to June 2009). Patients with high bifurcations identified at duplex scan were compared with a control group who had normal bifurcations. All patients were followed up at 1, 6 and 12 weeks post-operatively. The study endpoint was AVF patency. RESULTS: One hundred and five autologous BC AVF procedures were performed in our institution, of which 29 (27.6%) were identified as having a high brachial bifurcation on pre-operative duplex scan. The bifurcation was axillary in six patients and located at the proximal, middle and distal third of the humerus in nine, seven and seven patients, respectively. The actuarial functional patency rate was 53.2% (standard error = 9.6%) in the HB group and 76.2% (standard error = 4.9%) in the control group (log-rank test, p=0.027). CONCLUSIONS: These data show that aberrant brachial artery anatomy is both common (12%) and a predictor of autologous BC AVF failure. These data support the universal use of pre-AVF duplex scanning.


Assuntos
Derivação Arteriovenosa Cirúrgica/efeitos adversos , Artéria Braquial/cirurgia , Veias Braquiocefálicas/cirurgia , Oclusão de Enxerto Vascular/etiologia , Diálise Renal , Adulto , Idoso , Idoso de 80 Anos ou mais , Artéria Braquial/anormalidades , Artéria Braquial/diagnóstico por imagem , Artéria Braquial/fisiopatologia , Veias Braquiocefálicas/diagnóstico por imagem , Veias Braquiocefálicas/fisiopatologia , Distribuição de Qui-Quadrado , Feminino , Oclusão de Enxerto Vascular/diagnóstico por imagem , Oclusão de Enxerto Vascular/fisiopatologia , Humanos , Estimativa de Kaplan-Meier , Londres , Masculino , Pessoa de Meia-Idade , Guias de Prática Clínica como Assunto , Estudos Retrospectivos , Medição de Risco , Fatores de Risco , Fatores de Tempo , Falha de Tratamento , Ultrassonografia Doppler Dupla , Grau de Desobstrução Vascular
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