Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 44
Filtrar
Mais filtros










Base de dados
Intervalo de ano de publicação
2.
N Engl J Med ; 382(8): 697-705, 2020 02 20.
Artigo em Inglês | MEDLINE | ID: mdl-31860793

RESUMO

BACKGROUND: The causative agents for the current national outbreak of electronic-cigarette, or vaping, product use-associated lung injury (EVALI) have not been established. Detection of toxicants in bronchoalveolar-lavage (BAL) fluid from patients with EVALI can provide direct information on exposure within the lung. METHODS: BAL fluids were collected from 51 patients with EVALI in 16 states and from 99 healthy participants who were part of an ongoing study of smoking involving nonsmokers, exclusive users of e-cigarettes or vaping products, and exclusive cigarette smokers that was initiated in 2015. Using the BAL fluid, we performed isotope dilution mass spectrometry to measure several priority toxicants: vitamin E acetate, plant oils, medium-chain triglyceride oil, coconut oil, petroleum distillates, and diluent terpenes. RESULTS: State and local health departments assigned EVALI case status as confirmed for 25 patients and as probable for 26 patients. Vitamin E acetate was identified in BAL fluid obtained from 48 of 51 case patients (94%) in 16 states but not in such fluid obtained from the healthy comparator group. No other priority toxicants were found in BAL fluid from the case patients or the comparator group, except for coconut oil and limonene, which were found in 1 patient each. Among the case patients for whom laboratory or epidemiologic data were available, 47 of 50 (94%) had detectable tetrahydrocannabinol (THC) or its metabolites in BAL fluid or had reported vaping THC products in the 90 days before the onset of illness. Nicotine or its metabolites were detected in 30 of 47 of the case patients (64%). CONCLUSIONS: Vitamin E acetate was associated with EVALI in a convenience sample of 51 patients in 16 states across the United States. (Funded by the National Cancer Institute and others.).


Assuntos
Lesão Pulmonar Aguda/patologia , Líquido da Lavagem Broncoalveolar/química , Sistemas Eletrônicos de Liberação de Nicotina , Vaping/efeitos adversos , Vitamina E/análise , Lesão Pulmonar Aguda/etiologia , Adolescente , Adulto , Idoso , Fumar Cigarros , Óleo de Coco/análise , Feminino , Humanos , Limoneno/análise , Masculino , Pessoa de Meia-Idade , Estados Unidos , Adulto Jovem
3.
Retina ; 2019 Dec 19.
Artigo em Inglês | MEDLINE | ID: mdl-31860521

RESUMO

PURPOSE: Muller cells seem to be important in maintaining foveal morphology through connections between their foot processes and the internal limiting membrane (ILM). Internal limiting membrane peeling causes Muller cell trauma. We hypothesized that leaving a rim of unpeeled ILM around idiopathic macular holes undergoing vitrectomy surgery would improve postoperative foveal morphology and vision. METHODS: Prospective pilot study of fovea-sparing ILM peeling in a consecutive cohort of patients with macular holes over a 12-month period. Spectral-domain optical coherence tomography and Early Treatment Diabetic Retinopathy Study letters best-corrected visual acuity were assessed preoperatively and postoperatively, and foveal morphology and metamorphopsia postoperatively. The foveal sparing group was compared with a second consecutive cohort who received standard ILM peeling (control group). RESULTS: Thirty-four eyes of 34 patients were included in each group. Groups showed no significant preoperative differences. 34/34 holes were successfully closed with surgery in the foveal sparing group and 32/34 in the control group. The foveal sparing group showed better postoperative best-corrected visual acuity (67.7 vs. 63.8, P = 0.003) and best-corrected visual acuity improvement (25.1 vs. 20.2, P = 0.03). The foveal sparing group demonstrated thicker minimum foveal thickness (211 vs. 173 µm, P = 0.002) and less steep foveal depression (158 vs. 149, P = 0.002). CONCLUSION: Preserving nonpeeled ILM around macular holes resulted in a high closure rate, improved foveal morphology, and better postoperative best-corrected visual acuity. An appropriately powered randomized controlled study is warranted.

4.
MMWR Morb Mortal Wkly Rep ; 68(45): 1040-1041, 2019 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-31725707

RESUMO

CDC, the Food and Drug Administration (FDA), state and local health departments, and multiple public health and clinical partners are investigating a national outbreak of e-cigarette, or vaping, product use-associated lung injury (EVALI). Based on data collected as of October 15, 2019, 86% of 867 EVALI patients reported using tetrahydrocannabinol (THC)-containing products in the 3 months preceding symptom onset (1). Analyses of THC-containing product samples by FDA and state public health laboratories have identified potentially harmful constituents in these products, such as vitamin E acetate, medium chain triglyceride oil (MCT oil), and other lipids (2,3) (personal communication, D.T. Heitkemper, FDA Forensic Chemistry Center, November 2019). Vitamin E acetate, in particular, might be used as an additive in the production of e-cigarette, or vaping, products; it also can be used as a thickening agent in THC products (4). Inhalation of vitamin E acetate might impair lung function (5-7).


Assuntos
Líquido da Lavagem Broncoalveolar/química , Surtos de Doenças , Lesão Pulmonar/epidemiologia , Vaping/efeitos adversos , Adolescente , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estados Unidos/epidemiologia , Adulto Jovem
5.
Eur J Obstet Gynecol Reprod Biol ; 241: 94-98, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31479992

RESUMO

OBJECTIVE: To investigate the impact of the Episcissors-60 on obstetric anal sphincter injury (OASI) rates. STUDY DESIGN: Observational multi-centre time series analysis at four maternity units in the North-East of England. The main outcome measures were obstetric anal sphincter injury rates and delivery blood loss. RESULTS: Data were analysed for women who had a vaginal birth of a singleton pregnancy before (11,192) and after (8064) the introduction of the Episcissors-60. There were 2115 episiotomies before and 1498 after the introduction of the Episcissors-60, of which 1311 (87.5%) were undertaken with the Episcissors-60, 114 (7.6%) with other scissors and the scissors used were not stated in 73 (4.8%) women. There was no significant association between the introduction of Episcissors-60 and the performance of an episiotomy (χ2 = 0.006, p  = 0.94). Episiotomy was associated with a significant reduction in OASI rates (1.9% Vs 2.8%, odds ratio = 0.67 [0.51 - 0.86]; p =  0.001). There was no significant association between the introduction of the Episcissors-60 and the occurrence of OASIs in all women (χ2 = 0.6, p =  0.46) or in women who had an episiotomy (χ2 = 0.20, p = 0.71). In women who had an episiotomy, the mean estimated delivery blood loss was 550.3 ±â€¯8.2 ml before and 598.8 ±â€¯10.9 ml after the introduction of the Episcissors-60 (p <  0.001). CONCLUSION: Introduction of the Episcissors-60 was not associated with a change in OASI or episiotomy rates but may be associated with a small increase in delivery blood loss.


Assuntos
Canal Anal/lesões , Episiotomia/estatística & dados numéricos , Adulto , Episiotomia/efeitos adversos , Episiotomia/instrumentação , Feminino , Humanos , Gravidez , Adulto Jovem
6.
J Chromatogr B Analyt Technol Biomed Life Sci ; 1118-1119: 137-147, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-31035135

RESUMO

Progress toward better diagnosis and treatment of lipid metabolism-related diseases requires high throughput approaches for multiplexed quantitative analysis of structurally diverse lipids, including phospholipids (PLs). This work demonstrates a simplified "one-pot" phospholipid extraction protocol, as an alternative to conventional liquid-liquid extraction. Performed in a 96-well format, the extraction was coupled with high throughput UPLC and multiplexed tandem mass spectrometry (MS/MS) detection, allowing non-targeted quantification of phosphatidylcholines (PC), sphingomyelins (SM), lysophosphatidylcholines (LPC), phosphatidylethanolamines (PE), and phosphatidylinositols (PI). Using 50 µL aliquots of serum samples from 110 individuals, lipoproteins were fractionated by size, and analyzed for phospholipids and non-polar lipids including free cholesterol (FC), cholesteryl esters (CEs) and triglycerides (TGs). Analysis of serum samples with wide range of Total-TG levels showed significant differences in PL composition. The correlations of molar ratios in lipoprotein size fractions, SM/PL with FC/PL, PE/PL with TG/CE, and PE/PL with PI/PL, demonstrate the applicability of the method for quantitative composition analysis of high, low and very-low density lipoproteins (HDL, LDL and VLDL), and characterization of lipid metabolism related disease states.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Dislipidemias/sangue , Dislipidemias/diagnóstico , Fosfolipídeos/sangue , Espectrometria de Massas em Tandem/métodos , Humanos , Interações Hidrofóbicas e Hidrofílicas , Fosfolipídeos/isolamento & purificação , Reprodutibilidade dos Testes , Sensibilidade e Especificidade
7.
Health Res Policy Syst ; 17(1): 10, 2019 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-30691504

RESUMO

BACKGROUND: While the European Union is striving to become the 'Innovation Union', there remains a lack of quantifiable indicators to compare and benchmark regional innovation clusters. To address this issue, a HealthTIES (Healthcare, Technology and Innovation for Economic Success) consortium was funded by the European Union's Regions of Knowledge initiative, research and innovation funding programme FP7. HealthTIES examined whether the health technology innovation cycle was functioning differently in five European regional innovation clusters and proposed regional and joint actions to improve their performance. The clusters included BioCat (Barcelona, Catalonia, Spain), Medical Delta (Leiden, Rotterdam and Delft, South Holland, Netherlands), Oxford and Thames Valley (United Kingdom), Life Science Zürich (Switzerland), and Innova Észak-Alföld (Debrecen, Hungary). METHODS: Appreciation of the 'triple helix' of university-industry-government innovation provided the impetus for the development of two quantifiable innovation indexes and related indicators. The HealthTIES H-index is calculated for disease and technology platforms based on the h-index proposed by Hirsch. The HealthTIES Innovation Index is calculated for regions based on 32 relevant quantitative and discriminative indicators grouped into 12 categories and 3 innovation phases, namely 'Input' (n = 12), 'Innovation System' (n = 9) and 'Output' (n = 11). RESULTS: The HealthTIES regions had developed relatively similar disease and technology platform profiles, yet with distinctive strengths and weaknesses. The regional profiles of the innovation cycle in each of the three phases were surprisingly divergent. Comparative assessments based on the indicators and indexes helped identify and share best practice and inform regional and joint action plans to strengthen the competitiveness of the HealthTIES regions. CONCLUSION: The HealthTIES indicators and indexes provide useful practical tools for the measurement and benchmarking of university-industry-government innovation in European medical and life science clusters. They are validated internally within the HealthTIES consortium and appear to have a degree of external prima facie validity. Potentially, the tools and accompanying analyses can be used beyond the HealthTIES consortium to inform other regional governments, researchers and, possibly, large companies searching for their next location, analyse and benchmark 'triple helix' dynamics within their own networks over time, and to develop integrated public-private and cross-regional research and innovation strategies in Europe and beyond.


Assuntos
Benchmarking , Disciplinas das Ciências Biológicas , Pesquisa Biomédica , Governo , Indústrias , Universidades , Tecnologia Biomédica , Assistência à Saúde , Europa (Continente) , União Europeia , Humanos , Conhecimento , Tecnologia
8.
Retina ; 39(7): 1392-1398, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29517583

RESUMO

PURPOSE: To investigate the association between the vertical elevation of the external limiting membrane (ELM) and visual outcome in patients undergoing surgery for idiopathic full-thickness macular hole. METHODS: Retrospective observational study of a consecutive cohort of patients undergoing vitrectomy to treat macular hole. The greatest vertical height of the central ELM above the retinal pigment epithelium (ELM height) was measured on spectral domain optical coherence tomography preoperatively. The relationship of ELM height to other preoperative and postoperative variables, including macular hole width and height, and visual acuity was analyzed. RESULTS: Data from 91 eyes of 91 patients who had undergone successful hole closure were included. The mean ELM height was 220 µm (range 100-394). There were significant correlations between the ELM height and the diameter of the hole, hole height, and worsening preoperative visual acuity. For holes less than 400 µm in width, better postoperative visual acuity was significantly predicted by a lower ELM height. CONCLUSION: The ELM height varies widely in idiopathic macular hole. It is higher in eyes where the hole is wider and also when the hole itself is higher. For holes of less than 400 µm in width, a lower ELM height is a strong independent predictor of a good postoperative outcome.

9.
Ophthalmology ; 125(11): 1757-1764, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-29887331

RESUMO

PURPOSE: To identify the incidence, presenting features, treatment, and clinical course of optic disc pit maculopathy (ODPM) in the United Kingdom (UK). DESIGN: A 2-year nationwide prospective population-based study. SUBJECTS: All new incident cases of ODPM presenting to UK ophthalmologists using the British Ophthalmic Surveillance Unit monthly reporting system. METHODS: All reporting ophthalmologists were sent an initial questionnaire requesting data on previous medical and ophthalmic history, presentation details, investigation findings, and management. A further questionnaire was sent at 12 months post diagnosis to ascertain further outcome data. MAIN OUTCOME MEASURES: Visual acuity at initial presentation, at 1 year, and after any intervention. Foveal involvement and optical coherence tomography (OCT) findings, including retinal layers affected, and the location and size of the optic disc pit. Management, including observation, vitrectomy, and associated procedures. RESULTS: There were 74 confirmed new cases, giving an annual incidence of approximately 1 per 2 million. Complete data were available on 70 patients (70 eyes) at baseline and 68 after 1 year. There were 35 (50%) female patients with a mean age of 35 years (range, 3-82 years). Visual acuity at baseline ranged from 6/5 to hand movements. In 43 patients (61%) subretinal fluid (SRF) was present, whereas 27 (39%) had intraretinal fluid only. The presence of SRF was associated with worse vision and foveal involvement. Of the 53 eyes initially observed with 1-year follow-up, 10 (19%) deteriorated and 9 (16%) improved on OCT; eyes with SRF were more likely to worsen and those without SRF were more likely to improve. Fifteen of the 70 patients (21%) at baseline had primary surgery and a further 10 had deferred surgery within 1 year of presentation; 19 of these 25 eyes (76%) showed anatomic success with a dry fovea at 1 year of follow-up, and 15 (60%) had a greater than 0.1 logMAR improvement in visual acuity. CONCLUSION: The incidence and presenting features of ODPM were defined. Patients with SRF had worse vision and were more likely to deteriorate than patients with intraretinal fluid only. Surgery was anatomically successful in 75% of cases. Patients without SRF tended to remain stable with observation.


Assuntos
Anormalidades do Olho/epidemiologia , Disco Óptico/anormalidades , Doenças Retinianas/epidemiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Anormalidades do Olho/diagnóstico , Anormalidades do Olho/terapia , Feminino , Humanos , Incidência , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Vigilância em Saúde Pública , Doenças Retinianas/diagnóstico , Doenças Retinianas/terapia , Inquéritos e Questionários , Tomografia de Coerência Óptica , Reino Unido/epidemiologia
10.
PLoS One ; 13(4): e0194797, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29634782

RESUMO

Lipoproteins are complex molecular assemblies that are key participants in the intricate cascade of extracellular lipid metabolism with important consequences in the formation of atherosclerotic lesions and the development of cardiovascular disease. Multiplexed mass spectrometry (MS) techniques have substantially improved the ability to characterize the composition of lipoproteins. However, these advanced MS techniques are limited by traditional pre-analytical fractionation techniques that compromise the structural integrity of lipoprotein particles during separation from serum or plasma. In this work, we applied a highly effective and gentle hydrodynamic size based fractionation technique, asymmetric flow field-flow fractionation (AF4), and integrated it into a comprehensive tandem mass spectrometry based workflow that was used for the measurement of apolipoproteins (apos A-I, A-II, A-IV, B, C-I, C-II, C-III and E), free cholesterol (FC), cholesterol esters (CE), triglycerides (TG), and phospholipids (PL) (phosphatidylcholine (PC), sphingomyelin (SM), phosphatidylethanolamine (PE), phosphatidylinositol (PI) and lysophosphatidylcholine (LPC)). Hydrodynamic size in each of 40 size fractions separated by AF4 was measured by dynamic light scattering. Measuring all major lipids and apolipoproteins in each size fraction and in the whole serum, using total of 0.1 ml, allowed the volumetric calculation of lipoprotein particle numbers and expression of composition in molar analyte per particle number ratios. Measurements in 110 serum samples showed substantive differences between size fractions of HDL and LDL. Lipoprotein composition within size fractions was expressed in molar ratios of analytes (A-I/A-II, C-II/C-I, C-II/C-III. E/C-III, FC/PL, SM/PL, PE/PL, and PI/PL), showing differences in sample categories with combinations of normal and high levels of Total-C and/or Total-TG. The agreement with previous studies indirectly validates the AF4-LC-MS/MS approach and demonstrates the potential of this workflow for characterization of lipoprotein composition in clinical studies using small volumes of archived frozen samples.


Assuntos
Apolipoproteínas/sangue , Cromatografia Líquida/métodos , Fracionamento por Campo e Fluxo/métodos , Lipídeos/sangue , Lipoproteínas/sangue , Espectrometria de Massas em Tandem/métodos , Apolipoproteína A-I/metabolismo , Apolipoproteína B-100/metabolismo , Análise Química do Sangue/métodos , Calibragem , Colesterol/química , Humanos , Luz , Modelos Estatísticos , Tamanho da Partícula , Controle de Qualidade , Espalhamento de Radiação , Fluxo de Trabalho
11.
Methods Mol Biol ; 1722: 3-20, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29264795

RESUMO

The bacterial surfaceome, comprising outer membrane-sorted and/or associated (i.e., cell transporters), cell surface-exposed (i.e., adhesins) and extracellularly secreted proteins (i.e., toxins), has been characterized in bacterial pathogens, such as Bordetella pertussis (Bp) to provide information for use in development of diagnostic and prevention strategies. This protein subset has clinical significance, as these bacterial proteins are often associated with attachment to host cells, microbial pathogenesis and antibody-mediated immunity. Here we describe classical surface membrane protein enrichment techniques, followed by proteomic methodologies, such as gel-free protein separation and antibody-affinity capture technologies in combination with nano-liquid chromatography mass spectrometry, for the identification and characterization of Bp surfaceome proteins.


Assuntos
Antígenos de Bactérias/análise , Proteínas da Membrana Bacteriana Externa/análise , Bordetella pertussis/isolamento & purificação , Proteômica/métodos , Sequência de Aminoácidos , Afinidade de Anticorpos/imunologia , Antígenos de Bactérias/imunologia , Proteínas da Membrana Bacteriana Externa/imunologia , Bordetella pertussis/imunologia , Tampões (Química) , Carbonatos/química , Cromatografia Líquida , Bases de Dados de Proteínas , Imunoprecipitação/métodos , Espectrometria de Massas em Tandem
12.
J Cataract Refract Surg ; 43(9): 1135-1139, 2017 09.
Artigo em Inglês | MEDLINE | ID: mdl-28991608

RESUMO

PURPOSE: To evaluate the effect of a high-vacuum setting versus a low-vacuum setting on the efficiency of phacoemulsification. SETTING: Sunderland Eye Infirmary, Sunderland, United Kingdom. DESIGN: Prospective clinical trial. METHODS: Consecutive patients having cataract surgery in 2014 were recruited. Cataract surgery was performed by 2 experienced surgeons using a phacoemulsification machine with monitored forced infusion. The cataractous lens was split into 2 heminuclei using the stop-and-chop technique; in 1 heminucleus, phacoemulsification and aspiration used a high-vacuum setting (600 mm Hg; treatment group) and in the other heminucleus, a low-vacuum setting (350 mm Hg; control group). The high and low settings were alternated by case per the operating list to reduce surgeon bias. The main outcome measures were cumulative dissipated energy (CDE) and active heminucleus removal time. RESULTS: One hundred sixty patients (160 eyes) were enrolled in the study, and 158 were included in the analysis. The CDE per heminucleus was significantly lower with the high-vacuum setting than with the low-vacuum setting (mean 2.81 percent-seconds; 95% confidence interval (CI), 2.44-3.21 versus 3.81 percent-seconds; 95% CI, 3.38-4.20; P < .001). The active heminucleus removal time was significantly shorter in the high-vacuum group than the low-vacuum group (mean 27.77 seconds; 95% CI, 25.26-30.19 versus 33.59 seconds; 95% CI, 31.07-35.92; P < .001). The observed differences were independent of the surgeon, patient age and sex, incision size, and nucleus density. No intraoperative complications were observed in either group. CONCLUSION: A high-vacuum setting improved phacoemulsification efficiency using an active fluidics system and torsional phacoemulsification.


Assuntos
Facoemulsificação , Vácuo , Extração de Catarata , Humanos , Complicações Intraoperatórias , Implante de Lente Intraocular , Facoemulsificação/métodos , Estudos Prospectivos
13.
Proteomics Clin Appl ; 11(7-8)2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28296203

RESUMO

PURPOSE: Apolipoprotein A-I (ApoA-I) and apolipoprotein B-100 (ApoB-100) are amphipathic proteins that are strong predictors of cardiovascular disease risk. The traceable calibration of apolipoprotein assays is a persistent challenge, especially for ApoB-100, which cannot be solubilized in purified form. EXPERIMENTAL DESIGN: A simultaneous quantitation method for ApoA-I and ApoB-100 was developed using tryptic digestion without predigestion reduction and alkylation, followed by LC separation coupled with isotope dilution MS analysis. The accuracy of the method was assured by selecting structurally exposed signature peptides, optimal choice of detergent, protein:enzyme ratio, and incubation time. Peptide calibrators were value assigned by isobaric tagging isotope dilution MS amino acid analysis. RESULTS: The method reproducibility was validated in technical repeats of three serum samples, giving 2-3% intraday CVs (N = 5) and <7% interday CVs (N = 21). The repeated analysis of interlaboratory harmonization standards showed -1% difference for ApoA-I and -12% for ApoB-100 relative to the assigned value. The applicability of the method was demonstrated by repeated analysis of 24 patient samples with a wide range of total cholesterol and triglyceride levels. CONCLUSIONS AND CLINICAL RELEVANCE: The method is applicable for simultaneous analysis of ApoA-I and ApoB-100 in patient samples, and for characterization of serum pool calibrators for other analytical platforms.


Assuntos
Apolipoproteína A-I/química , Apolipoproteína B-100/química , Análise Química do Sangue/métodos , Espectrometria de Massas , Fragmentos de Peptídeos/sangue , Proteólise , Tripsina/metabolismo , Sequência de Aminoácidos , Apolipoproteína A-I/metabolismo , Apolipoproteína B-100/metabolismo , Calibragem , Cromatografia Líquida , Humanos , Isótopos/química , Modelos Lineares , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/metabolismo , Reprodutibilidade dos Testes
14.
J Proteomics ; 150: 258-267, 2017 01 06.
Artigo em Inglês | MEDLINE | ID: mdl-27667389

RESUMO

Apolipoproteins measured in plasma or serum are potential biomarkers for assessing metabolic irregularities that are associated with the development of cardiovascular disease (CVD). LC-MS/MS allows quantitative measurement of multiple apolipoproteins in the same sample run. However, the accuracy and precision of the LC-MS/MS measurement depends on the reproducibility of the enzymatic protein digestion step. With the application of an immobilized enzyme reactor (IMER), the reproducibility of the trypsin digestion can be controlled with high precision via flow rate, column volume and temperature. In this report, we demonstrate the application of an integrated IMER-LC-MS/MS platform for the simultaneous quantitative analysis of eight apolipoproteins. Using a dilution series of a characterized serum pool as calibrator, the method was validated by repeated analysis of pooled sera and individual serum samples with a wide range of lipid profiles, all showing intra-assay CV<4.4% and inter-assay CV<8%. In addition, the method was compared with traditional homogeneous digestion coupled LC-MS/MS for the quantification of apoA-I and apoB-100. Applied in large scale human population studies, this method can serve the translation of a wider panel of apolipoprotein biomarkers from research to clinical application. SIGNIFICANCE: Currently, the translation of apolipoprotein biomarkers to clinical application is impaired because of the high cost of large cohort studies using traditional single-analyte immunoassays. The application of on-line tryptic digestion coupled with LC-MS/MS analysis is an effective way to address this problem. In this work we demonstrate a high throughput, multiplexed, automated proteomics workflow for the simultaneous analysis of multiple proteins.


Assuntos
Apolipoproteínas/análise , Proteólise , Proteômica/métodos , Espectrometria de Massas em Tandem/métodos , Tripsina/metabolismo , Apolipoproteínas/metabolismo , Biomarcadores/sangue , Análise Química do Sangue/métodos , Proteínas Sanguíneas/análise , Proteínas Sanguíneas/metabolismo , Cromatografia Líquida , Estudos de Viabilidade , Humanos , Reprodutibilidade dos Testes
15.
Anal Bioanal Chem ; 409(5): 1379-1386, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27866257

RESUMO

Antibiotic resistance continues to contribute significantly to morbidity and mortality across the world. Developing new tests for antibiotic-resistant bacteria is a core action to combat resistant infections. We describe a method that uses phage amplification detection (PAD) combined with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) to rapidly identify Staphylococcus aureus and determine phenotypic susceptibility to cefoxitin. Samples tested for S. aureus are incubated together with bacteriophage in the presence and absence of cefoxitin and subjected to rapid trypsin digestion followed by MALDI-MS analysis. Tryptic peptides derived from amplified phage proteins can be detected by MALDI-MS, as validated by time-of-flight (TOF)/TOF analysis of each peptide combined with database searching. Methicillin-resistant S. aureus show significant phage amplification in the presence of cefoxitin, while methicillin-sensitive S. aureus show no phage amplification relative to a no-antibiotic control. We also show that PAD methodology can be implemented on an FDA-approved commercial MALDI-MS bacterial identification system to identify S. aureus and determine antibiotic susceptibility. The novelty of this assay includes the use of phage-derived tryptic peptides as detected by MALDI-MS to monitor the results of PAD on an instrument common to many modern microbiology laboratories.


Assuntos
Bacteriófagos/genética , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Testes de Sensibilidade Microbiana
16.
Genome Announc ; 4(6)2016 Dec 22.
Artigo em Inglês | MEDLINE | ID: mdl-28007855

RESUMO

Serum Institute of India is among the world's largest vaccine producers. Here, we report the complete genome sequences for four Bordetella pertussis strains used by Serum Institute of India in the production of whole-cell pertussis vaccines.

17.
Retina ; 36(1): 110-8, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26166800

RESUMO

PURPOSE: To determine if baseline fundoscopic and optical coherence tomography (OCT) features influence the clinical course of optic disk pit maculopathy. METHODS: A multicenter retrospective case note review was undertaken, using standardized OCT and clinical data collection. Visual success was defined as at least a two-line visual acuity improvement, anatomical success as full resolution of OCT foveal fluid with restoration of the normal foveal contour, and partial anatomical success as incomplete resolution of the OCT foveal fluid. Outcomes were compared with a synthesis of the literature, using similar eligibility criteria. RESULTS: Of 36 patients (36 eyes), 2 spontaneously improved and 34 underwent surgery. Visual success was achieved in 64% of surgical cases, anatomical success in 36%, and partial anatomical success in 47%. Cases with multilayer intraretinal and subretinal fluid were less likely to have visual success (P = 0.003). Cases where the fluid did not extend to the macular arcade vessels also had better visual and anatomical outcomes (P = 0.004 and 0.005, respectively). CONCLUSION: Fundoscopic and OCT features can help predict surgical outcome in optic disk pit maculopathy.


Assuntos
Anormalidades do Olho/diagnóstico , Disco Óptico/anormalidades , Doenças Retinianas/diagnóstico , Líquido Sub-Retiniano , Adolescente , Adulto , Criança , Anormalidades do Olho/cirurgia , Feminino , Seguimentos , Humanos , Masculino , Pessoa de Meia-Idade , Oftalmoscópios , Prognóstico , Doenças Retinianas/cirurgia , Estudos Retrospectivos , Tomografia de Coerência Óptica , Acuidade Visual/fisiologia , Vitrectomia
18.
Int J Proteomics ; 2015: 536537, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26090226

RESUMO

Bordetella pertussis (Bp) is the etiologic agent of pertussis (whooping cough), a highly communicable infection. Although pertussis is vaccine preventable, in recent years there has been increased incidence, despite high vaccine coverage. Possible reasons for the rise in cases include the following: Bp strain adaptation, waning vaccine immunity, increased surveillance, and improved clinical diagnostics. A pertussis outbreak impacted California (USA) in 2010; children and preadolescents were the most affected but the burden of disease fell mainly on infants. To identify protein biomarkers associated with this pertussis outbreak, we report a whole cellular protein characterization of six Bp isolates plus the pertussis acellular vaccine strain Bp Tohama I (T), utilizing gel-free proteomics-based mass spectrometry (MS). MS/MS tryptic peptide detection and protein database searching combined with western blot analysis revealed three Bp isolates in this study had markedly reduced detection of pertactin (Prn), a subunit of pertussis acellular vaccines. Additionally, antibody affinity capture technologies were implemented using anti-Bp T rabbit polyclonal antisera and whole cellular proteins to identify putative immunogens. Proteome profiling could shed light on pathogenesis and potentially lay the foundation for reduced infection transmission strategies and improved clinical diagnostics.

19.
Anal Chem ; 87(13): 6769-77, 2015 Jul 07.
Artigo em Inglês | MEDLINE | ID: mdl-26016659

RESUMO

The continued advance of antibiotic resistance in clinically relevant bacterial strains necessitates the development and refinement of assays that can rapidly and cost-effectively identify bacteria and determine their susceptibility to a panel of antibiotics. A methodology is described herein that exploits the specificity and physiology of the Staphylococci bacteriophage K to identify Staphylococcus aureus (S. aureus) and determine its susceptibility to clindamycin and cefoxitin. The method uses liquid chromatography-mass spectrometry to monitor the replication of bacteriophage after it is used to infect samples thought to contain S. aureus. Amplification of bacteriophage K indicates the sample contains S. aureus, for it is only in the presence of a suitable host that bacteriophage K can amplify. If bacteriophage amplification is detected in samples containing the antibiotics clindamycin or cefoxitin, the sample is deemed to be resistant to these antibiotics, respectively, for bacteriophage can only amplify in a viable host. Thus, with a single work flow, S. aureus can be detected in an unknown sample and susceptibility to clindamycin and cefoxitin can be ascertained. This Article discusses implications for the use of bacteriophage amplification in the clinical laboratory.


Assuntos
Antibacterianos/farmacologia , Bacteriófagos/genética , Espectrometria de Massas/métodos , Staphylococcus aureus/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Staphylococcus aureus/isolamento & purificação
20.
Ophthalmologica ; 233(3-4): 216-21, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25765054

RESUMO

AIMS: Analysis of pre-operative spectral domain optical coherence tomography (SD-OCT) characteristics of full-thickness macular holes (FTMH) and effect on optimum management. METHODS: We retrospectively reviewed SD-OCT characteristics of a consecutive cohort of patients waitlisted for FTMH surgery and categorized them by current evidence-based treatments. RESULTS: Out of the 106 holes analysed, 36 were small, 40 medium and 30 large. Initially, 33 holes had vitreomacular adhesion (VMA). 41 holes were analysed for change in characteristics with a median duration of 8 weeks between the scans. The number of small or medium holes decreased from 20 to 6 and that of large holes doubled. The number of holes with VMA halved. Smaller hole size (p = 0.014) and being phakic (p = 0.048) were associated with a larger increase in size. The strongest predictor of hole progression into a different surgical management category was the presence of VMA. CONCLUSION: FTMH characteristics can change significantly pre-operatively and affect optimal treatment choice.


Assuntos
Perfurações Retinianas/diagnóstico , Perfurações Retinianas/cirurgia , Vitrectomia , Idoso , Grupos Diagnósticos Relacionados , Progressão da Doença , Feminino , Humanos , Masculino , Período Pré-Operatório , Retina/metabolismo , Estudos Retrospectivos , Aderências Teciduais , Tomografia de Coerência Óptica , Acuidade Visual , Corpo Vítreo/metabolismo , Listas de Espera
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA