RESUMEN
The brain's response to acute injury is characterized by increased permeability of the blood-brain barrier (BBB) and pro-inflammatory microglia signaling, both of which have been linked to poor cognitive outcomes and neurological disease. The damaged BBB has increased leakiness, allowing serum proteins like fibrinogen into the brain, which interacts with local cells in a deleterious manner. At the same time, in response to injury, microglia demonstrate increased NLRP3 inflammasome activity and heightened release of pro-inflammatory cytokines. The relationship between increased fibrinogen uptake and microglial inflammasome signaling in the injured brain has not been well described. In this work, we investigate fibrinogen mediated NLRP3 inflammasome priming of BV-2 cells and primary adult microglia and propose a role for extracellular vesicles (EVs) as propagators of this interaction. Following exposure to fibrinogen microglia significantly upregulate transcription of IL-1ß, IL-6, NLRP3 and other pro-inflammatory cytokines which was sustained by repeated fibrinogen exposure. Inhibition of fibrinogen mediated NLRP3 signaling was achieved at the transcriptional and assembly level using cannabidiol (CBD) and the NLRP3 inhibitor MCC950, respectively. EVs released following NLRP3 priming carry IL-1ß, IL-18 mRNA and fibrinogen, propagate inflammatory signaling and can be detected in the circulation following BBB disruption in a preclinical stroke model. In conclusion, the interplay between fibrinogen extravasation, microglial NLRP3 signaling, and EV release can perpetuate chronic pro-inflammatory signaling and represents a novel method of inflammatory propagation.
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Vesículas Extracelulares , Inflamasomas , Humanos , Inflamasomas/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Microglía/metabolismo , Barrera Hematoencefálica/metabolismo , Fibrinógeno/metabolismo , Citocinas/metabolismo , Inflamación/metabolismo , Vesículas Extracelulares/metabolismoRESUMEN
Chronic microglia activation post-stroke is associated with worse neurological and cognitive outcomes. However, measurement of microglia activation in vivo is currently limited. Plasma derived extracellular vesicles (EVs) are cell-specific indicators that may allow for non-invasive measurement of microglia phenotype. The aim of this study was to identify activation-state specific microglia EVs (MEVs) in vitro followed by validation in an experimental stroke model. Following pro-inflammatory activation, MEVs contain the microglia protein TMEM119 alongside increased expression of the Toll-like receptor 4 co-receptor CD14. Immunoprecipitation followed by fluorescent nanoparticle tracking analysis (ONI Nanoimager) was used to confirm the isolation of TMEM119+/CD14+ EVs from rat plasma. Electron microscopy confirmed that TMEM119 and CD14 localize to the MEV membrane. To model ischemia, plasma was collected from 3-month wildtype Fischer344 rats prior to, 7 and 28 days after endothelin-1 or saline injection into the dorsal right striatum. Fluorescently labelled MEVs were directly measured in the plasma using nanoflow cytometry (Apogee A60 Microplus). We report a significant increase in circulating TMEM119+/CD14+ EVs 28-days post-stroke in comparison to baseline levels and saline-injected rats, which correlated weakly with stroke volume. TMEM119+/MHC-II+ EVs were also increased post-stroke in comparison to baseline and saline-injected animals. This study is the first to describe an EV biomarker of activated microglia detected directly in plasma following stroke and represents a future tool for the measurement of microglia activity in vivo.
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Vesículas Extracelulares , Microglía , Accidente Cerebrovascular , Animales , Ratas , Biomarcadores , Cuerpo Estriado , FenotipoRESUMEN
Gangliosides have a wide variety of biological functions due to their location on the outer leaflet of plasma membranes. They form a critical component of membrane rafts, or ganglioside-enriched microdomains, where they influence the physical properties of the membrane as well as its function. Gangliosides can change their structure to meet their external and internal environmental demands. This ability to change structure makes gangliosides both fascinating and technologically challenging targets to identify and understand. A full understanding on how gangliosides are regulated within the central nervous system (CNS) is critical, as ganglioside dysregulation is observed in the aging brain as well as in several neurodegenerative injuries and diseases such as stroke, Alzheimer's disease, Parkinson's disease, Huntington's disease and several lysosomal storage disorders diseases, including Tay Sach's disease. Mass spectrometry (MS) has become a useful means to better understand ganglioside composition and function. Imaging mass spectrometry (IMS) provides the added benefit of placing analytical information within an anatomical context. This review article will discuss recent advances in MS-based detection methods, with a focus on IMS-based approaches to help understand the spatial-specific role gangliosides in the healthy brain as in CNS injuries and disease.
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Encéfalo/metabolismo , Gangliósidos/análisis , Envejecimiento , Enfermedad de Alzheimer/metabolismo , Enfermedad de Alzheimer/patología , Animales , Gangliósidos/química , Humanos , Enfermedad de Huntington/metabolismo , Enfermedad de Huntington/patología , Espectrometría de Masas , Enfermedad de Parkinson/metabolismo , Enfermedad de Parkinson/patología , Accidente Cerebrovascular/metabolismo , Accidente Cerebrovascular/patologíaRESUMEN
Background and Aims: Ephemeral seagrasses that respond rapidly to environmental changes are important marine habitats. However, they are under threat due to human activity and are logistically difficult and expensive to study. This study aimed to develop a new functional-structural environmentally dependent model of ephemeral seagrass, able to integrate our understanding of ephemeral seagrass growth dynamics and assess options for potential management interventions, such as seagrass transplantation. Methods: A functional-structural plant model was developed in which growth and senescence rates are mechanistically linked to environmental variables. The model was parameterized and validated for a population of Halophila stipulacea in the Persian Gulf. Key Results: There was a good match between empirical and simulated results for the number of apices, net rhizome length or net number of internodes using a 330 d simulation. Simulated data were more variable than empirical data. Simulated structural patterns of seagrass rhizome growth qualitatively matched empirical observations. Conclusions: This new model successfully simulates the environmentally dependent growth and senescence rates of our case-study ephemeral seagrass species. It produces numerical and visual outputs that help synthesize our understanding of how the influence of environmental variables on plant functional processes affects overall growth patterns. The model can also be used to assess the potential outcomes of management interventions like seagrass transplantation, thus providing a useful management tool. It is freely available and easily adapted for new species and locations, although validation with more species and environments is required.
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Hydrocharitaceae/fisiología , Modelos Biológicos , Biomasa , Ecosistema , Ambiente , Hydrocharitaceae/anatomía & histología , Hydrocharitaceae/crecimiento & desarrollo , Océano Índico , Biología Marina , Rizoma/anatomía & histología , Rizoma/crecimiento & desarrollo , Rizoma/fisiología , Estaciones del Año , Especificidad de la EspecieRESUMEN
BACKGROUND AND OBJECTIVES: The type and clinical characteristics of patients identified with commonly used definitions of massive transfusion (MT) are largely unknown. The objective of this study was to define the clinical characteristics of patients meeting different definitions of MT for the purpose of patient recruitment in observational studies. MATERIALS AND METHODS: Data were extracted on all patients who received red blood cell (RBC) transfusions in 2010 at three tertiary Australian hospitals. MT patients were identified according to three definitions: ≥10 units RBC in 24 h (10/24 h), ≥6 units RBC in 6 h (6/6 h) and ≥5 units RBC in 4 h (5/4 h). Clinical coding data were used to assign bleeding context. Data on in-hospital mortality were also extracted. RESULTS: Five hundred and forty-two patients met at least one MT definition, with 236 (44%) included by all definitions. The most inclusive definition was 5/4 h (508 patients, 94%) followed by 6/6 h (455 patients, 84%) and 10/24 h (251 patients, 46%). Importantly, 40-55% of most types of critical bleeding events and 82% of all obstetric haemorrhage cases were excluded by the 10/24 h definition. Patients who met both the 5/4 h and 10/24 h definitions were transfused more RBCs (19 vs. 8 median total RBC units; P < 0·001), had longer ventilation time (120 vs. 55 h; P < 0·001), median ICU (149 vs. 99 h; P < 0·001) and hospital length of stay (23 vs. 18 h; P = 0·006) and had a higher in-hospital mortality rate (23·3% vs. 16·4%; P = 0·050). CONCLUSION: The 5/4 h MT definition was the most inclusive, but combination with the 10/24 h definition appeared to identify a clinically important patient cohort.
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Transfusión de Eritrocitos/estadística & datos numéricos , Transfusión de Eritrocitos/normas , Hemorragia/epidemiología , Hemorragia/terapia , Mortalidad Hospitalaria , Adulto , Anciano , Australia/epidemiología , Transfusión de Eritrocitos/mortalidad , Femenino , Humanos , Estimación de Kaplan-Meier , Masculino , Persona de Mediana EdadRESUMEN
All four serotypes of dengue virus (DENV) cause the full spectrum of disease. Therefore, vaccines must protect against all serotypes. To evaluate candidate vaccines, a human challenge model of dengue serotype 3 (rDEN30Δ30) was developed. All challenge virus recipients safely met the primary endpoint of viremia and secondary endpoints of rash and seroconversion to DENV-3.
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La Crosse virus (LACV) and Jamestown Canyon virus (JCV), family Bunyaviridae, are mosquito-borne viruses that are endemic in North America and recognized as etiologic agents of encephalitis in humans. Both viruses belong to the California encephalitis virus serogroup, which causes 70 to 100 cases of encephalitis a year. As a first step in creating live attenuated viral vaccine candidates for this serogroup, we have generated a recombinant LACV expressing the attachment/fusion glycoproteins of JCV. The JCV/LACV chimeric virus contains full-length S and L segments derived from LACV. For the M segment, the open reading frame (ORF) of LACV is replaced with that derived from JCV and is flanked by the untranslated regions of LACV. The resulting chimeric virus retained the same robust growth kinetics in tissue culture as observed for either parent virus, and the virus remains highly infectious and immunogenic in mice. Although both LACV and JCV are highly neurovirulent in 21 day-old mice, with 50% lethal dose (LD50) values of 0.1 and 0.5 log10 PFU, respectively, chimeric JCV/LACV is highly attenuated and does not cause disease even after intracerebral inoculation of 10³ PFU. Parenteral vaccination of mice with 10¹ or 10³ PFU of JCV/LACV protected against lethal challenge with LACV, JCV, and Tahyna virus (TAHV). The chimeric virus was infectious and immunogenic in rhesus monkeys and induced neutralizing antibodies to JCV, LACV, and TAHV. When vaccinated monkeys were challenged with JCV, they were protected against the development of viremia. Generation of highly attenuated yet immunogenic chimeric bunyaviruses could be an efficient general method for development of vaccines effective against these pathogenic viruses.
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Quimera/inmunología , Virus de la Encefalitis de California/genética , Encefalitis de California/prevención & control , Expresión Génica , Virus La Crosse/inmunología , Proteínas del Envoltorio Viral/inmunología , Animales , Anticuerpos Antivirales/inmunología , Línea Celular , Quimera/genética , Quimera/crecimiento & desarrollo , Encefalitis de California/inmunología , Encefalitis de California/virología , Humanos , Virus La Crosse/genética , Virus La Crosse/crecimiento & desarrollo , Macaca mulatta , Ratones , Vacunación , Proteínas del Envoltorio Viral/genética , Vacunas Virales/genética , Vacunas Virales/inmunologíaRESUMEN
OBJECTIVES: Errors in administration of blood products can lead to poor patient outcomes including fatal ABO incompatible transfusions. This pilot study sought to establish whether the use of two-dimensional (2D) barcode technology combined with patient identification software designed to assist in blood administration improves the bedside administration of transfusions in an Australian tertiary hospital. STUDY DESIGN AND METHODS: The study was conducted in a Haematology/Oncology Day Clinic of a major metropolitan hospital, to evaluate the use of 2D barcode technology and patient safety-software and hand-held PDAs to assist nursing staff in patient identification and blood administration. Comparative audits were conducted before and after the technology's implementation. RESULTS: The preimplementation transfusion practice audits demonstrated a poor understanding of the blood checking process, with focus on the product rather than patient identification. Following the implementation of 2D barcode technology and patient safety-software, there was significant improvement in administration practice. Positive, verbal patient identification improved from 57% (51/90) to 94% (75/80). Similarly, the cross-referencing of the patient's identification with the patient's wristband improved from 36% (32/90) to 94% (75/80), and the cross-referencing of patient ID on the compatibility tag to wristbands improved from 48% (43/90) to 99% (79/80). Importantly, the 2D barcode technology and patient safety-software saw 100% (80/80) of checks being conducted at the patient bedside, compared with 76% (68/90) in the preimplementation audits. CONCLUSION: This pilot study demonstrates that 2D barcode technology and patient safety-software significantly improves the bedside check of patient and blood product identification in an Australian setting.
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Incompatibilidad de Grupos Sanguíneos/prevención & control , Transfusión Sanguínea , Sistemas de Registros Médicos Computarizados/instrumentación , Seguridad del Paciente , Programas Informáticos , Centros de Atención Terciaria , Australia , Auditoría Clínica , Femenino , Humanos , Masculino , Proyectos PilotoRESUMEN
Little is known about the evolution, diversity, and functional significance of secondary metabolites in reproductive plant parts, particularly fruits and seeds of plants in natural ecosystems. We compared the concentration and diversity of amides among six tissue types of Piper reticulatum: leaves, roots, flowers, unripe fruit pulp, ripe fruit pulp, and seeds. This represents the first detailed description of amides in P. reticulatum, and we identified 10 major and 3 minor compounds using GC/MS and NMR analysis. We also detected 30 additional unidentified minor amide components, many of which were restricted to one or a few plant parts. Seeds had the highest concentrations and the highest diversity of amides. Fruit pulp had intermediate concentrations and diversity that decreased with ripening. Leaves and roots had intermediate concentrations, but the lowest chemical diversity. In addition, to investigate the potential importance of amide concentration and diversity in plant defense, we measured leaf herbivory and seed damage in natural populations, and examined the relationships between amide occurrence and plant damage. We found no correlations between leaf damage and amide diversity or concentration, and no correlation between seed damage and amide concentration. The only relationship we detected was a negative correlation between seed damage and amide diversity. Together, our results provide evidence that there are strong selection pressures for fruit and seed defense independent of selection in vegetative tissues, and suggest a key role for chemical diversity in fruit-frugivore interactions.
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Amidas/análisis , Frutas/química , Piper , Amidas/metabolismo , Flores/química , Flores/metabolismo , Frutas/metabolismo , Herbivoria , Piper/metabolismo , Hojas de la Planta/química , Hojas de la Planta/metabolismo , Raíces de Plantas/química , Raíces de Plantas/metabolismoRESUMEN
Ischemic stroke affects millions of individuals worldwide and a high prevalence of survivors experience cognitive deficits. At present, the underlying mechanisms that drive post-stroke cognitive decline are not well understood. Microglia play a critical role in the post-stroke inflammatory response, but experimental studies show that an accumulation of chronically activated microglia can be harmful and associates with cognitive impairment. This study assessed the effect of acute post-stroke minocycline treatment on chronic microglia and astrocyte expression within the infarct and remote white matter regions, as well as its effect on various domains of cognitive function post-stroke. Nine-month-old male rats received an injection of endothelin-1 into the right dorsal striatum to induce transient focal ischemia, and then were treated with minocycline or saline for 4 days post-stroke. Rats were tested using a series of lever-pressing tasks and the Morris water maze to assess striatal-based learning, cognitive flexibility, and spatial learning and reference memory. We found that minocycline-treated rats had smaller stroke-induced infarcts and less microglia activation in the infarct area and remote white matter regions compared to saline-treated rats at 28 days post-stroke. The behavioural testing results differed according to the cognitive domain; whereas minocycline-treated rats trended towards improved striatal-based learning in a lever-pressing task, but cognitive flexibility was unaffected during the subsequent set-shifting task. Furthermore, minocycline treatment unexpectedly impaired spatial learning, yet it did not alter reference memory. Collectively, we show that post-stroke minocycline treatment can reduce chronic microglia activation even in remote brain regions, with domain-specific effects on cognitive function.
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Interactions between plants and herbivores are central in most ecosystems, but their strength is highly variable. The amount of variability within a system is thought to influence most aspects of plant-herbivore biology, from ecological stability to plant defense evolution. Our understanding of what influences variability, however, is limited by sparse data. We collected standardized surveys of herbivory for 503 plant species at 790 sites across 116° of latitude. With these data, we show that within-population variability in herbivory increases with latitude, decreases with plant size, and is phylogenetically structured. Differences in the magnitude of variability are thus central to how plant-herbivore biology varies across macroscale gradients. We argue that increased focus on interaction variability will advance understanding of patterns of life on Earth.
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Variación Biológica Poblacional , Herbivoria , Defensa de la Planta contra la Herbivoria , Plantas , Ecosistema , Filogenia , Animales , Evolución BiológicaRESUMEN
BACKGROUND: Prothrombinex-VF (a three-factor prothrombin complex concentrate) contains little factor VII. Therefore, the Warfarin Reversal Consensus Guidelines from 2004 published by The Australasian Society of Haemostasis and Thrombosis recommend that it be administered with fresh frozen plasma to reverse warfarin anticoagulation. AIM: To evaluate the efficacy and safety of Prothrombinex-VF used alone in warfarin reversal. METHODS: Adult patients requiring urgent reversal of warfarin anticoagulation were defined as having achieved complete (target international normalized ratio (INR) <1.4) or partial reversal (target INR 1.4-2.0) of their anticoagulation. Prothrombinex-VF was given at doses of between 25 and 50 IU/kg based on the intent of reversal and an INR was obtained 30min post infusion. RESULTS: A total of 50 patients (mean age 72years, range 32-85years) was included. The median initial INR in the complete reversal arm (n= 35) was 3.5 (range 1.7-20) with 91% achieving the target INR (mean 1.1, range 0.9-1.4). In the partial reversal arm (n= 15) the mean initial INR was 5.6 (range 2.5-12) with 93% achieving the target INR (mean 1.6, range 1.4-2.2). There were no adverse effects attributed to Prothrombinex-VF. CONCLUSIONS: Prothrombinex-VF is very effective and safe when used alone to reverse warfarin anticoagulation. The supplementary use of fresh frozen plasma in these patients is not required. A review of the current Warfarin Reversal Consensus Guidelines is needed.
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Anticoagulantes/efectos adversos , Factores de Coagulación Sanguínea/administración & dosificación , Coagulación Sanguínea/efectos de los fármacos , Relación Normalizada Internacional , Warfarina/efectos adversos , Adulto , Anciano , Anciano de 80 o más Años , Coagulación Sanguínea/fisiología , Estudios de Cohortes , Femenino , Humanos , Relación Normalizada Internacional/normas , Persona de Mediana Edad , Estudios Prospectivos , Factores de TiempoRESUMEN
OBJECTIVES: To assess the incidence, prevalence and consequences of illness in one professional academy rugby league club during an in-season period. DESIGN: Observational prospective cohort study. METHOD: Seventeen male rugby league players (age 17.7±0.7 years, stature 178.8±5.1cm, body mass 87.2±9.6kg) completed a weekly self-report illness questionnaire using an amended version of the Oslo Sports Trauma Research Centre (OSTRC) questionnaire on health problems. RESULTS: A total of 24 new illnesses were reported over the 25-week study period. 65% of players experienced at least one illness during the study. The incidence of illness in this cohort was 14.3 per 1000-player days, with the respiratory system being most commonly affected (n=15; 62.5%). The average weekly illness prevalence was 10.3%. Time-loss illness incidence was 1.4 per 1000-player days. Loss of body mass and sleep disruptions were the most commonly reported consequences of illness episodes. Mean body mass loss during a period of illness was 2.2±0.6kg. CONCLUSIONS: Academy rugby league players are most commonly affected by respiratory illness with a total of nineteen training and competition days lost to illness. Associated consequences of illness, such as loss of body mass and sleep disruptions may present a challenge and negatively impact a rugby league player's development. Appropriate medical provisions should be provided for Academy rugby league players to support them during periods of illness to limit the impact of these consequences.
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Enfermedad , Fútbol Americano , Adolescente , Costo de Enfermedad , Enfermedades Gastrointestinales/epidemiología , Humanos , Incidencia , Masculino , Prevalencia , Estudios Prospectivos , Enfermedades Respiratorias/epidemiología , Autoinforme , Enfermedades de la Piel/epidemiología , Sueño , Encuestas y CuestionariosRESUMEN
A general protocol for rigorous evaluation of diagnostic systems in medicine was applied successfully in a comparative study of two radiologic techniques. Accuracies of computed tomography and radionuclide scanning in detecting, localizing, and diagnosing brain lesions were assessed with a sample of patients in whom tumor had been suspected. The principal means of analysis was the "relative operating characteristic," which is unique in providing a measure of accuracy that is largely independent of decision biases. Computed tomography was found to be substantially more accurate than radionuclide scanning.
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Encéfalo/diagnóstico por imagen , Tomografía Computarizada por Rayos X , Encefalopatías/diagnóstico por imagen , Neoplasias Encefálicas/diagnóstico por imagen , Diagnóstico Diferencial , Reacciones Falso Negativas , Reacciones Falso Positivas , Humanos , CintigrafíaRESUMEN
Histological studies have demonstrated that polycystic ovaries (PCO) contain increased numbers of preantral follicles with a specific increase in primary follicles. Polycystic ovary syndrome is associated with hyperandrogenism and pre- and postnatal androgenization of primates increases the pool of growing follicles producing changes resembling PCO. In vitro studies could test the hypothesis that androgens alter early folliculogenesis, but conventional culture techniques for small follicles are generally unsuitable in non-rodent species. Our objective was to develop and use a method to investigate the effects of testosterone on early folliculogenesis. We adapted an in ovo technique in which lamb cortical ovarian fragments were grafted onto the chorioallantoic membrane of fertilised chick eggs. Optimal experimental conditions for vascularisation and survival of tissue were determined and the model then used to investigate the effects of testosterone on follicle growth. Eggs were inoculated with testosterone at the time of implantation of the ovarian tissue, which was retrieved 5 days later. Tissue was sectioned and follicles staged and counted. There was no wholesale initiation of primordial follicle growth over the 5-day in ovo culture. Importantly, the proportion of primordial, primary and secondary follicles remained similar to those in unimplanted tissue. Testosterone increased the number of primary follicles by 50% compared with controls, an effect that was largely due to a reduction in atresia. In conclusion, incubation of ovarian cortex with testosterone reproduces the changes in early folliculogenesis reported in histological studies of PCO.
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Folículo Ovárico/efectos de los fármacos , Síndrome del Ovario Poliquístico/metabolismo , Testosterona/farmacología , Animales , Embrión de Pollo , Membrana Corioalantoides/metabolismo , Femenino , Atresia Folicular/efectos de los fármacos , Modelos Animales , Ovario/metabolismo , Ovario/trasplante , Óvulo , Ovinos , Estimulación Química , Trasplante HeterólogoRESUMEN
Carmarthen Bay, UK, regularly supports internationally important numbers (>16,000) of non-breeding Common Scoters Melanitta nigra. The spill of 72,000 tonnes of crude oil from the Sea Empress in 1996 affected birds both through direct mortality and likely pollution of key food resources. Numbers were greatly reduced following the spill, whilst changes in the distribution of birds within Carmarthen Bay suggested that potentially sub-optimal foraging zones were used. However, ten years after the incident, numbers of Common Scoter were no different to those recorded immediately before the spill. Compared to some other spills, rapid revival is evident. Numbers increased to pre-spill levels within three winters and distributional changes suggested a concurrent return to previously contaminated feeding areas, implying that the ecosystem had regenerated sufficiently to support its top predator. The importance of prolonged, standardised monitoring of bird numbers and distribution as indicators of ecological recovery from environmental damage is emphasised.
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Accidentes , Anseriformes , Petróleo , Contaminantes Químicos del Agua , Animales , Monitoreo del Ambiente , Densidad de Población , Dinámica Poblacional , Navíos , GalesAsunto(s)
Artefactos , Recolección de Muestras de Sangre/métodos , Errores Diagnósticos/prevención & control , Ácido Edético/sangre , Contaminación de Equipos/prevención & control , Flebotomía/métodos , Potasio/sangre , Adolescente , Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Adulto JovenRESUMEN
There is a lack of published studies on laundering in ambulance services. We performed bacterial culture on soiled and unsoiled uniforms and reusable mop heads artificially contaminated with Escherichia coli, Staphylococcus aureus, and Clostridium difficile spores. Current laundering processes used for routine cleans in the ambulances appears, from our simulations, to be effective at reducing vegetative pathogenic bacteria to undetectable levels, <3.398log10 colony-forming units (S. aureus and E. coli). Reduced levels of C. difficile were still detected after laundering but the risk this poses for infection is unknown, as background levels of these spores in the environment are unknown.