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1.
Br J Anaesth ; 120(2): 291-298, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29406178

RESUMO

BACKGROUND: It is unclear if isolated postoperative cardiac-troponin elevation, often referred to as myocardial injury, represents a pathological event, as control studies in otherwise healthy adults are lacking. METHODS: In this single-centre prospective observational cohort study, serial high-sensitivity cardiac troponin T (hscTnT) plasma concentrations were obtained from young, healthy adults undergoing elective orthopaedic surgery at three time points: before operation, 2-6 h, and 18-30 h after surgery. End points were hscTnT increases after surgery: ≥20% (exceeding analytical variability), ≥50% (exceeding short-term biological variability), and ≥85% (exceeding long-term biological variability). The secondary end point was myocardial injury, defined as new postoperative hscTnT elevation >99th % upper reference limit (URL) (women >10 ng litre-1; men >15 ng litre-1). RESULTS: Amongst the study population (n=95), no hscTnT increase ≥20% was detected in 68 patients (73%). A hscTnT increase between 20% and 49% was observed in 17 patients (18%), 50-84% in seven patients (7%), and ≥85% in three patients (3%). Twenty patients (21%) had an absolute ΔhscTnT between 0 and 2 ng litre-1, 12 patients (13%) between 2 and 4 ng litre-1, three patients between 4 and 6 ng litre-1, and one patient (1%) between 6 and 8 ng litre-1. Myocardial injury (new hscTnT elevation >99th%) was diagnosed in one patient (1%). The median hscTnT concentrations did not increase after operation, and were 4 (3.9-5, inter-quartile range) ng litre-1 at baseline, 4 (3.9-5) ng litre-1 at 2-6 h after surgery, and 4 (3.9-5) ng litre-1 on postoperative day 1. CONCLUSIONS: One in four young adult patients without known cardiovascular disease developed a postoperative hscTnT increase, but without exceeding the 99th% URL and without evidence of myocardial ischaemia. These results may have important ramifications for the concept of postoperative myocardial injury, as they suggest that, in some patients, postoperative cardiac-troponin increases may be the result of a normal physiological process in the surgical setting. CLINICAL TRIAL REGISTRATION: NCT 02394288.


Assuntos
Troponina T/sangue , Adulto , Biomarcadores/sangue , Eletrocardiografia , Feminino , Humanos , Masculino , Procedimentos Ortopédicos , Complicações Pós-Operatórias/diagnóstico , Complicações Pós-Operatórias/terapia , Estudos Prospectivos , Resultado do Tratamento , Adulto Jovem
2.
Epidemiol Infect ; 143(9): 1964-71, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25387485

RESUMO

Blood culture contamination (BCC) has been associated with unnecessary antibiotic use, additional laboratory tests and increased length of hospital stay thus incurring significant extra hospital costs. We set out to assess the impact of a staff educational intervention programme on decreasing intensive care unit (ICU) BCC rates to <3% (American Society for Microbiology standard). BCC rates during the pre-intervention period (January 2006-May 2011) were compared with the intervention period (June 2011-December 2012) using run chart and regression analysis. Monthly ICU BCC rates during the intervention period were reduced to a mean of 3.7%, compared to 9.5% during the baseline period (P < 0.001) with an estimated potential annual cost savings of about £250,100. The approach used was simple in design, flexible in delivery and efficient in outcomes, and may encourage its translation into clinical practice in different healthcare settings.


Assuntos
Coleta de Amostras Sanguíneas/normas , Sangue/microbiologia , Pessoal de Saúde/educação , Testes Hematológicos/normas , Competência Clínica , Reações Falso-Positivas , Humanos , Irlanda do Norte , Estudos Prospectivos , Estudos Retrospectivos
3.
Epidemiol Infect ; 142(3): 494-500, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23735079

RESUMO

The objective of this study was to evaluate the impact of restricting high-risk antibiotics on methicillin-resistant Staphylococcus aureus (MRSA) incidence rates in a hospital setting. A secondary objective was to assess the impact of reducing fluoroquinolone use in the primary-care setting on MRSA incidence in the community. This was an interventional, retrospective, ecological investigation in both hospital and community (January 2006 to June 2010). Segmented regression analysis of interrupted time-series was employed to evaluate the intervention. The restriction of high-risk antibiotics was associated with a significant change in hospital MRSA incidence trend (coefficient=-0·00561, P=0·0057). Analysis showed that the intervention relating to reducing fluoroquinolone use in the community was associated with a significant trend change in MRSA incidence in community (coefficient=-0·00004, P=0·0299). The reduction in high-risk antibiotic use and fluoroquinolone use contributed to both a reduction in incidence rates of MRSA in hospital and community (primary-care) settings.


Assuntos
Antibacterianos/uso terapêutico , Uso de Medicamentos/normas , Infecções Estafilocócicas/tratamento farmacológico , Cefalosporinas/uso terapêutico , Clindamicina/uso terapêutico , Infecção Hospitalar/tratamento farmacológico , Infecção Hospitalar/epidemiologia , Fluoroquinolonas/uso terapêutico , Humanos , Incidência , Staphylococcus aureus Resistente à Meticilina , Irlanda do Norte/epidemiologia , Atenção Primária à Saúde , Estudos Retrospectivos , Infecções Estafilocócicas/epidemiologia
4.
Epidemiol Infect ; 142(2): 404-8, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23657218

RESUMO

The objective of this study was to evaluate the effect of age-adjusted comorbidity and alcohol-based hand rub on monthly hospital antibiotic usage, retrospectively. A multivariate autoregressive integrated moving average (ARIMA) model was built to relate the monthly use of all antibiotics grouped together with age-adjusted comorbidity and alcohol-based hand rub over a 5-year period (April 2005-March 2010). The results showed that monthly antibiotic use was positively related to the age-adjusted comorbidity index (concomitant effect, coefficient 1·103, P = 0·0002), and negatively related to the use of alcohol-based hand rub (2-month delay, coefficient -0·069, P = 0·0533). Alcohol-based hand rub is considered a modifiable factor and as such can be identified as a target for quality improvement programmes. Time-series analysis may provide a suitable methodology for identifying possible predictive variables that explain antibiotic use in healthcare settings. Future research should examine the relationship between infection control practices and antibiotic use, identify other infection control predictive factors for hospital antibiotic use, and evaluate the impact of enhancing different infection control practices on antibiotic use in a healthcare setting.


Assuntos
Antibacterianos/uso terapêutico , Higiene das Mãos/estatística & dados numéricos , Higienizadores de Mão/uso terapêutico , Hospitais/estatística & dados numéricos , Adulto , Fatores Etários , Idoso , Infecções Bacterianas/tratamento farmacológico , Infecções Bacterianas/epidemiologia , Infecções Bacterianas/prevenção & controle , Comorbidade , Infecção Hospitalar/tratamento farmacológico , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/prevenção & controle , Humanos , Pessoa de Meia-Idade , Estudos Retrospectivos
5.
Int J Lab Hematol ; 35(1): 77-81, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22938565

RESUMO

INTRODUCTION: With proper logistical support and sponsorship, a laboratory in an industrialized nation might be able to act as a reference laboratory for clinicians based in a developing country. METHODS: We built on previous experience in the clinical laboratory to see whether a specialized histopathology service (hematopathology) could be provided to a developing country without the expertise or experience to do it in country. RESULTS: Over an 13-year period, 582 cases from 579 individuals were analyzed. Principal pathologic findings included acute leukemia in 84 cases (14%), dyspoiesis in one or more of the hematopoietic lineages in 65 cases (11%, including three cases with high-grade myelodysplasia), 23 cases (4%) with findings suspicious for a chronic myeloproliferative disorder, 35 cases (6%) with findings suspicious for a lymphoproliferative disorder, and infectious organisms (presumably Leishmania in most instances) in 9 (1%) of cases. Specimens from 45 cases (8%) were unsatisfactory owing to extreme hemodilution and/or specimen degeneration. CONCLUSION: With proper support, a medical laboratory in an industrialized nation may serve as a reference facility for a developing nation. The use of existing infrastructure may be remarkably effective to achieve optimal turnaround time. Although the lack of ancillary studies and follow-up biopsies limit the ability to achieve a definitive diagnosis in many cases, this must be viewed in the context of the limited ability to diagnose or manage hematopoietic neoplasia in developing nations.


Assuntos
Exame de Medula Óssea , Neoplasias Hematológicas/diagnóstico , Testes Hematológicos , Cooperação Internacional , Leishmaniose/diagnóstico , Aeronaves , Medula Óssea/patologia , Exame de Medula Óssea/economia , Exame de Medula Óssea/normas , Países Desenvolvidos , Países em Desenvolvimento , Eritreia , Custos de Cuidados de Saúde , Neoplasias Hematológicas/sangue , Neoplasias Hematológicas/patologia , Testes Hematológicos/economia , Testes Hematológicos/normas , Hematologia/economia , Hematologia/métodos , Hematologia/organização & administração , Humanos , Infectologia/economia , Infectologia/métodos , Infectologia/organização & administração , Agências Internacionais , Leishmaniose/sangue , Leishmaniose/parasitologia , Leishmaniose/patologia , Oncologia/economia , Oncologia/métodos , Oncologia/organização & administração , Patologia Clínica/economia , Patologia Clínica/métodos , Patologia Clínica/organização & administração , Manejo de Espécimes , Telecomunicações , Fatores de Tempo , Estados Unidos , Instituições Filantrópicas de Saúde
6.
Epidemiol Infect ; 140(9): 1714-20, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22115422

RESUMO

The objective of this research was to assess current patterns of hospital antibiotic prescribing in Northern Ireland and to determine targets for improving the quality of antibiotic prescribing. A point prevalence survey was conducted in four acute teaching hospitals. The most commonly used antibiotics were combinations of penicillins including ß-lactamase inhibitors (33·6%), metronidazole (9·1%), and macrolides (8·1%). The indication for treatment was recorded in 84·3% of the prescribing episodes. A small fraction (3·9%) of the surgical prophylactic antibiotic prescriptions was for >24 h. The results showed that overall 52·4% of the prescribed antibiotics were in compliance with the hospital antibiotic guidelines. The findings identified the following indicators as targets for quality improvement: indication recorded in patient notes, the duration of surgical prophylaxis and compliance with hospital antibiotic guidelines. The results strongly suggest that antibiotic use could be improved by taking steps to address the identified targets for quality improvement.


Assuntos
Antibacterianos/administração & dosagem , Prescrições de Medicamentos , Revisão de Uso de Medicamentos , Idoso , Idoso de 80 Anos ou mais , Prescrições de Medicamentos/normas , Prescrições de Medicamentos/estatística & dados numéricos , Feminino , Fidelidade a Diretrizes/estatística & dados numéricos , Pesquisas sobre Atenção à Saúde , Hospitais de Ensino , Humanos , Masculino , Pessoa de Meia-Idade , Irlanda do Norte
7.
J Hosp Infect ; 77(3): 233-6, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21216032

RESUMO

Blood cultures have an important role in the diagnosis of serious infections, although contamination of blood cultures (i.e. false-positive blood cultures) is a common problem within the hospital setting. The objective of the present investigation was to determine the impact of the false-positive blood culture results on the following outcomes: length of stay, hotel costs, antimicrobial costs, and costs of laboratory and radiological investigation. A retrospective case-control study design was used in which 142 false-positive blood culture cases were matched with suitable controls (patients for whom cultures were reported as true negatives). The matching criteria included age, comorbidity score and month of admission to the hospital. The research covered a 13-month period (July 2007 to July 2008). The findings indicated that differences in means, between cases and controls, for the length of hospital stay and the total costs were 5.4 days [95% CI (confidence interval): 2.8-8.1 days; P<0.001] and £5,001.5 [$7,502.2; 95% CI: £3,283.9 ($4,925.8) to £6,719.1 ($10,078.6); P<0.001], respectively. Consequently, and considering that 254 false-positive blood cultures had occurred in the study site hospital over a one-year period, patients with false-positive blood cultures added 1372 extra hospital days and incurred detrimental additional hospital costs of £1,270,381 ($1,905,572) per year. The findings therefore demonstrate that false-positive blood cultures have a significant impact on increasing hospital length of stay, laboratory and pharmacy costs. These findings highlight the need to intervene to raise the standard of blood-culture-taking technique, thus improving both the quality of patient care and resource use.


Assuntos
Sangue/microbiologia , Infecção Hospitalar/economia , Meios de Cultura/economia , Contaminação de Equipamentos/economia , Custos Hospitalares , Adulto , Idoso , Antibacterianos/economia , Antibacterianos/uso terapêutico , Técnicas Bacteriológicas/economia , Coleta de Amostras Sanguíneas/métodos , Estudos de Casos e Controles , Análise Custo-Benefício , Infecção Hospitalar/tratamento farmacológico , Reações Falso-Positivas , Feminino , Hospitais , Humanos , Tempo de Internação/economia , Masculino , Pessoa de Meia-Idade , Adulto Jovem
11.
J Hosp Infect ; 71(1): 22-8, 2009 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19036472

RESUMO

Rapid detection of MRSA may be important for the control of MRSA spread in hospitals. The aim of this investigation was to compare the use of a rapid polymerase chain reaction (PCR) screening method with standard culture for the detection of meticillin-resistant Staphylococcus aureus (MRSA) colonisation and to determine its impact on the incidence of MRSA in two hospital wards. During the first phase of the investigation (four months), patients in a surgical ward were screened using the rapid PCR technique and patients in a medical/cardiology ward were screened with standard culture methods. During the second phase of the investigation (four months), MRSA screening methods were switched between the two wards. An audit of infection control practices on each ward was made at the end of each phase in order to check whether any changes had occurred that might influence the risks of MRSA transmission. Use of the rapid PCR method significantly reduced the median time between swabs being taken, to the results being telephoned to the wards (excluding weekends), from 47 to 21 h (P<0.001). However, comparison of MRSA incidence during use of PCR (20/1000 bed-days) and culture methods (22.1/1000 bed-days) revealed no significant difference in incidence on the surgical ward (P=0.69). Regarding the medical/cardiology ward, analysis of data was complicated by an increase in the detection of MRSA during the PCR phase (P<0.05). The study demonstrated that rapid PCR can significantly reduce the turnaround times but reducing the time between swabs being taken to results being telephoned to the ward is still not sufficient to limit the transmission of MRSA.


Assuntos
Infecção Hospitalar/prevenção & controle , Controle de Infecções/métodos , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Infecções Estafilocócicas/prevenção & controle , Técnicas de Tipagem Bacteriana , Infecção Hospitalar/diagnóstico , Humanos , Reação em Cadeia da Polimerase , Infecções Estafilocócicas/diagnóstico , Fatores de Tempo
12.
J Clin Pharm Ther ; 29(3): 257-62, 2004 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-15153087

RESUMO

OBJECTIVE: The present study addresses pharmacy expenditure within a surgical directorate in a UK hospital. The aim of the study was to develop a health care resource group (HRG)-based costing model that can be used to forecast pharmacy expenditure based on surgical casemix. Such a model will be of benefit as an expenditure projection tool at a time when hospitals are developing accelerated operation programmes in an attempt to decrease hospital waiting times. METHOD: During the period February-April 2000, nursing staff recorded all pharmacy sourced items for each individual operation in the theatres used for general surgery, ENT surgery and gynaecological procedures; each operation was also classified according to its HRG. The associated costs of the items per HRG were identified and the average pharmaceutical cost per HRG calculated and included in the costing model. The model derived costs over the study period were compared with the actual pharmacy expenditure which was obtained from the pharmacy computer system. Finally HRG data for operations carried out in February 2002 were costed using the model for validation purposes. RESULTS: The estimated pharmaceutical cost for surgery items for February-April 2000 was 121,235 UK pounds. This figure was 3.92% over the actual pharmaceutical expenditure as determined from computer records. The February 2002 casemix varied considerably from that of 2000. However, the model estimated pharmaceutical cost of surgery performed in February 2002 (38,054 UK pounds) was again very similar to the computer logged expenditure (1.09% under the actual expenditure for that period) indicating the robustness of the HRG-based costing approach.


Assuntos
Custos de Medicamentos , Pesquisas sobre Atenção à Saúde/economia , Procedimentos Cirúrgicos Operatórios/economia , Alocação de Custos/métodos , Análise Custo-Benefício/métodos , Coleta de Dados/métodos , Atenção à Saúde/tendências , Pesquisas sobre Atenção à Saúde/métodos , Necessidades e Demandas de Serviços de Saúde/economia , Necessidades e Demandas de Serviços de Saúde/tendências , Custos Hospitalares , Hospitais de Ensino , Humanos , Modelos Econômicos , Procedimentos Cirúrgicos Operatórios/classificação , Procedimentos Cirúrgicos Operatórios/estatística & dados numéricos , Fatores de Tempo , Listas de Espera
13.
Br J Clin Pharmacol ; 53(2): 163-71, 2002 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-11851640

RESUMO

AIMS: Helicobacter pylori (H. pylori) eradication rate varies according to the treatment regimen used and other factors, e.g. antimicrobial resistance and patient compliance. The aim of the present study was to evaluate the influence of patient counselling and follow-up on H. pylori eradication rates and to document the effectiveness of a 1 week eradication regimen consisting of lansoprazole (30 mg once daily), amoxicillin (1 g twice daily) and clarithromycin (500 mg twice daily). METHODS: Seventy-six dyspeptic patients, who at endoscopy were found to have gastritis, duodenitis or ulceration, and a positive H. pylori urease test, were recruited. Patients were randomly assigned to an intervention group (n = 38) or a control group (n = 38). Intervention patients received their medicines via the hospital pharmacy and were counselled (and followed up) by a hospital pharmacist. Control patients were given a standard advice sheet and referred to their GP who prescribed the same therapy. RESULTS: Intervention patients exhibited a statistically significant improvement in the H. pylori eradication rate (94.7% vs 73.7%; P = 0.02) and compliance (92.1% vs 23.7; P < 0.001). Of the 64 H. pylori eradicated patients, 62 were able to eliminate their antisecretory medication compared with only 12 of the H. pylori persistent patients (P < 0.001). A pharmacoeconomic evaluation indicated that counselling and follow-up reduced the direct costs of eradication by approximately 30 UK pounds per patient. CONCLUSIONS: Structured patient counselling and follow-up can have a significant effect on H. pylori eradication rates and should be a routine part of therapy.


Assuntos
Aconselhamento , Infecções por Helicobacter/prevenção & controle , Helicobacter pylori , Cooperação do Paciente , Adulto , Idoso , Idoso de 80 Anos ou mais , Continuidade da Assistência ao Paciente , Custos e Análise de Custo , Feminino , Infecções por Helicobacter/tratamento farmacológico , Infecções por Helicobacter/economia , Humanos , Masculino , Pessoa de Meia-Idade , Inquéritos e Questionários , Resultado do Tratamento
16.
Clin Lab Med ; 21(2): 269-84, 2001 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-11396083

RESUMO

Advancing technology has allowed for the migration of laboratory testing from the central laboratory to the near-patient setting, leading ideally to a shorter therapeutic turnaround time. This potential benefit, however, comes with a price tag. Assessing the cost-benefit ratio on a per test basis cannot effectively be done in a generalized manner, because each hospital must evaluate this with respect to its own unique circumstances. There are, however, certain outcomes, such as decreased LOS and decreased blood-product usage that, if achieved, far outweigh the cost of POCT, justifying its use. Any hospital attempting to implement POCT must also realize that hospital operations are affected by such use and that adjustments and careful laboratory oversight are required.


Assuntos
Custos Hospitalares/estatística & dados numéricos , Laboratórios Hospitalares/economia , Ciência de Laboratório Médico/economia , Sistemas Automatizados de Assistência Junto ao Leito/economia , Custos e Análise de Custo , Humanos , Garantia da Qualidade dos Cuidados de Saúde/economia , Estados Unidos
17.
Clin Chem ; 47(5): 919-25, 2001 May.
Artigo em Inglês | MEDLINE | ID: mdl-11325897

RESUMO

BACKGROUND: (99m)Tc-sestamibi scans and rapid, intraoperative intact parathyroid hormone (PTH) assays allow preoperative identification of diseased glands and intraoperative confirmation of diseased gland removal, respectively. Use of these two new technologies may facilitate simpler, more concise surgery, shorter hospital stays, and decreased costs for frozen-section analysis. One major drawback to this new strategy has been the high cost of rapid point-of-care PTH assays. METHODS: We performed rapid PTH assays with the DPC Turbo PTH assay on the DPC IMMULITE automated analyzer. The number of intraoperative frozen sections, type of anesthesia, surgical approach, length of hospital stay, and pre- and postoperative calcium values were compared between a group of 49 patients undergoing parathyroidectomy where the intraoperative PTH assay was used in conjunction with preoperative imaging, and a historical control group of 55 patients before the use of these two technologies in our institution. RESULTS: Comparison of the Turbo PTH assay to the standard IMMULITE PTH assay gave the following: y = 1.08 x - 4.36 (r = 0.97; n = 48). For the 49 patients, the median turnaround time for each intraoperative PTH determination was 19 min (range, 14-40 min). The median decrease in PTH values from baseline was 88% (range, 33-99%). Thirty-seven patients required two PTH determinations, 7 required three, 4 had four, and 1 required five determinations. The average laboratory cost for the rapid intraoperative PTH assays was < $100 per patient (range, $55 to $113). Compared with the control group, the experimental group had significantly fewer frozen sections (1.4 vs 2.5; P < 0.0001), shorter hospital stays (17 discharged on the day of surgery vs none discharged on the day of surgery; P < 0.0001), greater use of local anesthesia (33% vs 0%; P < 0.001), and more unilateral, rather than bilateral neck explorations (65% vs 0%; P < 0.001). CONCLUSIONS: The combination of intraoperative Turbo PTH assay and preoperative (99m)Tc-sestamibi scans can lead to significant decreases in laboratory and surgical pathology costs, hospital stays, and exposure to general anesthesia by facilitating concise parathyroidectomy surgery.


Assuntos
Glândulas Paratireoides/diagnóstico por imagem , Glândulas Paratireoides/cirurgia , Hormônio Paratireóideo/sangue , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anestesia/métodos , Cálcio/metabolismo , Feminino , Secções Congeladas , Humanos , Hiperparatireoidismo/diagnóstico por imagem , Hiperparatireoidismo/economia , Hiperparatireoidismo/cirurgia , Imunoensaio , Período Intraoperatório , Tempo de Internação , Medições Luminescentes , Masculino , Pessoa de Meia-Idade , Paratireoidectomia/métodos , Sistemas Automatizados de Assistência Junto ao Leito , Cintilografia , Compostos Radiofarmacêuticos , Tecnécio Tc 99m Sestamibi
19.
Infect Immun ; 69(4): 2172-9, 2001 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-11254572

RESUMO

This study demonstrates that pretreatment of macrophages with phosphatidylinositol, of either soya bean or mycobacterial origin, results in a down-regulation of the binding and uptake of Mycobacterium tuberculosis by the phagocytes. We also describe the novel observation that cardiolipin induces an increase in the binding and uptake of M. tuberculosis by macrophages. Neither phospholipid interacts with macrophages via the 2F8 epitope of scavenger receptor A, and treatment of macrophages with either phospholipid results in a down-regulation of CR3 function and tumor necrosis factor alpha production by the phagocyte. We have also shown that the ability of macrophages to interact with mycobacteria is greatly affected by an as yet unidentified product from the interaction of chloroform and polypropylene tubes.


Assuntos
Aderência Bacteriana/efeitos dos fármacos , Cardiolipinas/farmacologia , Macrófagos/microbiologia , Mycobacterium tuberculosis/fisiologia , Fosfatidilinositóis/farmacologia , Animais , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Fator de Necrose Tumoral alfa/biossíntese
20.
Biochemistry ; 39(47): 14504-14, 2000 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-11087404

RESUMO

Three structural variants (PV5, PV7, and PV8) of the horseshoe crab cationic antimicrobial peptide polyphemusin I were designed with improved amphipathic profiles. Circular dichroism spectroscopy analysis indicated that in phosphate buffer polyphemusin I, PV7, and PV8 displayed the spectrum of a type II beta-turn-rich structure, but, like polyphemusin I, all three variants adopted a typical beta-sheet structure in an anionic lipid environment. Both polyphemusin I and variants were potent broad spectrum antimicrobials that were clearly bactericidal at their minimal inhibitory concentrations. The variants were moderately less active in vitro but more effective in animal models. Moreover, these variants exhibited delayed bacterial killing, whereas polyphemusin I killed Escherichia coli UB1005 within 5 min at 2.5 microg/mL. All the peptides showed similar abilities to bind to bacterial lipopolysaccharide (LPS) and permeabilize bacterial outer membranes. Consistent with this was the observation that all peptides significantly inhibited cytokine production by LPS-stimulated macrophages and penetrated polyanionic LPS monolayers to similar extents. None of the peptides had affinity for neutral lipids as evident from both tryptophan fluorescence spectroscopy and Langmuir monolayer analysis. As compared to polyphemusin I, all variants showed reduced ability to interact with anionic lipids, and the hemolytic activity of the variants was decreased by 2-4-fold. In contrast, polyphemusin I efficiently depolarized the cytoplasmic membrane of E. coli, as assessed using a membrane potential sensitive fluorescent dye 3,3-dipropylthiacarbocyanine (diSC(3)5) assay, but the variants showed a substantially delayed and decreased depolarizing ability. The coincident assessment of cell viability indicated that depolarization of the bacterial cytoplasmic membrane potential by polyphemusin I occurred prior to lethal damage to cells. Our data suggest that increase of amphipathicity of beta-sheet polyphemusin I generally resulted in variants with decreased activity for membranes. Interestingly, all variants showed an improved ability to protect mice both against infection by Pseudomonas aeruginosa and from endotoxaemia.


Assuntos
Anti-Infecciosos/química , Peptídeos Catiônicos Antimicrobianos/química , Proteínas da Membrana Bacteriana Externa/química , Lipopolissacarídeos/química , Fosfolipídeos/química , Homologia de Sequência de Aminoácidos , Animais , Anti-Infecciosos/administração & dosagem , Anti-Infecciosos/síntese química , Anti-Infecciosos/farmacologia , Peptídeos Catiônicos Antimicrobianos/administração & dosagem , Peptídeos Catiônicos Antimicrobianos/síntese química , Peptídeos Catiônicos Antimicrobianos/farmacologia , Proteínas da Membrana Bacteriana Externa/metabolismo , Permeabilidade da Membrana Celular/efeitos dos fármacos , Dicroísmo Circular , Proteínas de Ligação a DNA/síntese química , Proteínas de Ligação a DNA/química , Modelos Animais de Doenças , Endotoxinas/antagonistas & inibidores , Feminino , Proteínas Hemolisinas/farmacologia , Caranguejos Ferradura , Humanos , Injeções Intraperitoneais , Lipopolissacarídeos/metabolismo , Lipossomos/química , Lipossomos/metabolismo , Camundongos , Testes de Sensibilidade Microbiana , Neutropenia/microbiologia , Neutropenia/mortalidade , Neutropenia/prevenção & controle , Peptídeos Cíclicos/síntese química , Peptídeos Cíclicos/química , Ligação Proteica/efeitos dos fármacos , Engenharia de Proteínas , Isoformas de Proteínas/síntese química , Isoformas de Proteínas/química , Isoformas de Proteínas/farmacologia , Estrutura Secundária de Proteína , Relação Estrutura-Atividade
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