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1.
Nature ; 588(7838): 436-441, 2020 12.
Artículo en Inglés | MEDLINE | ID: mdl-33328667

RESUMEN

Rivers support some of Earth's richest biodiversity1 and provide essential ecosystem services to society2, but they are often fragmented by barriers to free flow3. In Europe, attempts to quantify river connectivity have been hampered by the absence of a harmonized barrier database. Here we show that there are at least 1.2 million instream barriers in 36 European countries (with a mean density of 0.74 barriers per kilometre), 68 per cent of which are structures less than two metres in height that are often overlooked. Standardized walkover surveys along 2,715 kilometres of stream length for 147 rivers indicate that existing records underestimate barrier numbers by about 61 per cent. The highest barrier densities occur in the heavily modified rivers of central Europe and the lowest barrier densities occur in the most remote, sparsely populated alpine areas. Across Europe, the main predictors of barrier density are agricultural pressure, density of river-road crossings, extent of surface water and elevation. Relatively unfragmented rivers are still found in the Balkans, the Baltic states and parts of Scandinavia and southern Europe, but these require urgent protection from proposed dam developments. Our findings could inform the implementation of the EU Biodiversity Strategy, which aims to reconnect 25,000 kilometres of Europe's rivers by 2030, but achieving this will require a paradigm shift in river restoration that recognizes the widespread impacts caused by small barriers.


Asunto(s)
Ecosistema , Ríos , Agricultura/estadística & datos numéricos , Altitud , Biodiversidad , Conjuntos de Datos como Asunto , Restauración y Remediación Ambiental/métodos , Restauración y Remediación Ambiental/tendencias , Europa (Continente) , Actividades Humanas , Humanos , Modelos Logísticos , Aprendizaje Automático , Densidad de Población , Centrales Eléctricas/provisión & distribución
2.
BMC Med Educ ; 23(1): 866, 2023 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-37968656

RESUMEN

BACKGROUND: Imperial College School of Medicine, London UK, introduced a new curriculum in 2019, with a focus on the GMC outcomes for graduates, and pedagogy best practice. The new curriculum included formative assessments, named engagement and feedback assessments (EFAs), to support learning, and attainment in the summative examinations. The aims of this study were to assess the validity of EFAs and to determine whether they have utility as a modified form of programmatic assessment to inform decision-making regarding possible interventions by measuring and analysing attendance at and performance in these formative events. METHODS: Seven hundred and sixty-one students were included in the study and assessment results were included for academic years 2019/20 to 2020/21. Forty-one data points per student, (27 in Year 1 and 14 in Year 2) were used, to compare EFA scores with the summative performance. Attendance was monitored through engagement with the EFAs. RESULTS: Cohort 1 (enrolled 2019): In year 1, EFAs were associated with summative exam scores (overall r = 0.63, p < 0.001). Year 2, EFA scores were also associated with summative scores (overall r = 0.57, p < 0.001), including the clinical practical assessment (r = 0.45, p < 0.001). Missing two or more EFAs was associated with a significant increase in the likelihood of failing one or more summative examinations in the first year (OR: 7.97, 95% CI 2.65-34.39) and second year (OR: 3.20, 95% CI 1.74-5.95). Missing more than two EFAs in their first year was also associated with a higher risk of failing a summative examination in the second year (OR: 2.47, 95% CI 1.33-4.71). Students who increased their attendance between year 1 and 2 fared better in summative assessment than those who maintained poor attendance, whereas those that reduced their attendance fared worse than those that maintained high attendance. Cohort 2 (enrolled 2020): Analysis of cohort 2 supported these findings and in this cohort missing two or more EFAs was again associated with an increased likelihood of failing a summative examination (OR = 4.00, 95% CI = 2.02-7.90). CONCLUSION: Our EFA model has validity in predicting performance in summative assessments and can inform prospective interventions to support students' learning. Enhancing attendance and engagement can improve outcomes.


Asunto(s)
Educación de Pregrado en Medicina , Estudiantes de Medicina , Humanos , Retroalimentación , Evaluación Educacional/métodos , Estudios Prospectivos , Educación de Pregrado en Medicina/métodos
3.
J Pharmacol Exp Ther ; 376(1): 51-63, 2021 01.
Artículo en Inglés | MEDLINE | ID: mdl-33115824

RESUMEN

Asthma is still an incurable disease, and there is a recognized need for novel small-molecule therapies for people with asthma, especially those poorly controlled by current treatments. We previously demonstrated that calcium-sensing receptor (CaSR) negative allosteric modulators (NAMs), calcilytics, uniquely suppress both airway hyperresponsiveness (AHR) and inflammation in human cells and murine asthma surrogates. Here we assess the feasibility of repurposing four CaSR NAMs, which were originally developed for oral therapy for osteoporosis and previously tested in the clinic as a novel, single, and comprehensive topical antiasthma therapy. We address the hypotheses, using murine asthma surrogates, that topically delivered CaSR NAMs 1) abolish AHR; 2) are unlikely to cause unwanted systemic effects; 3) are suitable for topical application; and 4) inhibit airway inflammation to the same degree as the current standard of care, inhaled corticosteroids, and, furthermore, inhibit airway remodeling. All four CaSR NAMs inhibited poly-L-arginine-induced AHR in naïve mice and suppressed both AHR and airway inflammation in a murine surrogate of acute asthma, confirming class specificity. Repeated exposure to inhaled CaSR NAMs did not alter blood pressure, heart rate, or serum calcium concentrations. Optimal candidates for repurposing were identified based on anti-AHR/inflammatory activities, pharmacokinetics/pharmacodynamics, formulation, and micronization studies. Whereas both inhaled CaSR NAMs and inhaled corticosteroids reduced airways inflammation, only the former prevented goblet cell hyperplasia in a chronic asthma model. We conclude that inhaled CaSR NAMs are likely a single, safe, and effective topical therapy for human asthma, abolishing AHR, suppressing airways inflammation, and abrogating some features of airway remodeling. SIGNIFICANCE STATEMENT: Calcium-sensing receptor (CaSR) negative allosteric modulators (NAMs) reduce airway smooth muscle hyperresponsiveness, reverse airway inflammation as efficiently as topical corticosteroids, and suppress airway remodeling in asthma surrogates. CaSR NAMs, which were initially developed for oral therapy of osteoporosis proved inefficacious for this indication despite being safe and well tolerated. Here we show that structurally unrelated CaSR NAMs are suitable for inhaled delivery and represent a one-stop, steroid-free approach to asthma control and prophylaxis.


Asunto(s)
Antiasmáticos/uso terapéutico , Asma/tratamiento farmacológico , Indanos/uso terapéutico , Naftalenos/uso terapéutico , Fenilpropionatos/uso terapéutico , Quinazolinonas/uso terapéutico , Receptores Sensibles al Calcio/agonistas , Regulación Alostérica , Animales , Antiasmáticos/efectos adversos , Antiasmáticos/farmacología , Bronquios/efectos de los fármacos , Bronquios/metabolismo , Reposicionamiento de Medicamentos , Células HEK293 , Humanos , Indanos/efectos adversos , Indanos/farmacología , Masculino , Ratones , Ratones Endogámicos BALB C , Naftalenos/efectos adversos , Naftalenos/farmacología , Fenilpropionatos/efectos adversos , Fenilpropionatos/farmacología , Quinazolinonas/efectos adversos , Quinazolinonas/farmacología , Receptores Sensibles al Calcio/metabolismo
4.
Cytotherapy ; 23(2): 111-118, 2021 02.
Artículo en Inglés | MEDLINE | ID: mdl-33246883

RESUMEN

BACKGROUND: Cell replacement therapy (CRT) for Huntington disease (HD) requires a source of striatal (STR) progenitors capable of restoring the function lost due to STR degeneration. Authentic STR progenitors can be collected from the fetal putative striatum, or whole ganglionic eminence (WGE), but these tissues remain impractical for widespread clinical application, and alternative donor sources are required. Here we begin exploring the possibility that induced pluripotent stem cells (iPSC) derived from WGE may retain an epigenetic memory of their tissue of origin, which could enhance their ability to differentiate into STR cells. RESULTS: We generate four iPSC lines from human WGE (hWGE) and establish that they have a capacity similar to human embryonic stem cells with regard to their ability to differentiate toward an STR phenotype, as measured by expression and demethylation of key STR genes, while maintaining an overall different methylome. Finally, we demonstrate that these STR-differentiated hWGE iPSCs share characteristics with hWGE (i.e., authentic STR tissues) both in vitro and following transplantation into an HD model. Overall, iPSCs derived from human WGE show promise as a donor source for CRT for HD.


Asunto(s)
Tratamiento Basado en Trasplante de Células y Tejidos , Cuerpo Estriado , Enfermedad de Huntington , Células Madre Pluripotentes Inducidas , Diferenciación Celular , Cuerpo Estriado/citología , Humanos , Enfermedad de Huntington/terapia
5.
J Acoust Soc Am ; 150(5): 3874, 2021 11.
Artículo en Inglés | MEDLINE | ID: mdl-34852591

RESUMEN

Acoustic bubble curtains have been marketed as relatively low cost and easily maintained behavioural deterrents for fisheries management. Their energy efficiency can be improved by reducing air flow and exploiting bubble resonance. In a series of three flume experiments, we: (1) investigated the reactions of carp to a low air flow bubble curtain, (2) compared the effectiveness of resonant versus non-resonant insonified bubble curtains (for the same volume flux of gas injected through the nozzles) to deter passage, and determined the stimuli responsible for eliciting deterrence, and (3) included the effect of visual cues generated by the bubble curtain. This study showed that bubble curtains with a higher proportion of resonant bubbles deterred carp relatively better. Passage rejection was likely influenced by multiple cues at distances within a body length of the fish- specifically the rate of change in both particle motion and flow velocity caused by rising bubbles. All acoustic bubble curtains were less effective in the presence of daylight, suggesting that vision plays an important role at mediating carp reactions. We discuss the importance of ascertaining the bubble size distribution, in addition to the gas flow rate and aperture size, when characterising acoustically active bubble curtains.


Asunto(s)
Carpas , Acústica , Animales , Movimiento (Física)
6.
J Exp Biol ; 223(Pt 17)2020 09 01.
Artículo en Inglés | MEDLINE | ID: mdl-32748794

RESUMEN

One of the most detrimental factors in the drastic decline of the critically endangered European eel (Anguilla anguilla) was the inadvertent introduction of the invasive nematode Anguillicoloides crassus Infection primarily affects the swimbladder, a gas-filled organ that enables the eel to control its depth in the water. A reduction in swimbladder function may be fatal for eel undergoing their spawning migration to the Sargasso Sea, a journey of over 5000 km. Although the physiological damage caused by this invasive parasite is well studied through the use of quantifiable gross pathological indices, providing a good measure of the swimbladder health status, they cannot separate the role of mechanical and morphological damage. Our study examined the appropriateness of three commonly used indices as a measure of mechanical damage by performing uniaxial tensile tests on swimbladder specimens obtained from an infected eel population. When the test results were compared with the gross pathological indices it was found that thickness correlated most strongly with mechanical damage, both confirming and, more importantly, explaining the counterintuitive findings of earlier work. In a damaged swimbladder, the immune response leads to a trade-off; increasing wall thickness raises the pressure required for organ rupture but decreases strength. The results indicate that for moderate infection the mechanical integrity of the swimbladder can be maintained. For severe infection, however, a reduction in mechanical integrity may reach a tipping point, thereby affecting the successful completion of their oceanic migration.


Asunto(s)
Anguilla , Dracunculoidea , Enfermedades de los Peces , Infecciones por Nematodos , Sacos Aéreos , Animales , Infecciones por Nematodos/veterinaria
7.
Clin Nephrol ; 94(3): 105-115, 2020 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-32631484

RESUMEN

Premature muscle tiring (fatigue) is an important complication of advanced kidney disease, and results in a reduced ability to walk and exercise. As patients move from pre-dialysis care to dialysis, they begin to lose their residual kidney function and experience muscle wasting. Not all patients on dialysis are affected equally by the loss of muscle mass, and the effect of dialysis mode (peritoneal dialysis and hemodialysis) on muscle wasting is not known. This review will explore molecular mechanisms relevant to muscle wasting in end-stage kidney disease and in particular address some important insights into the emerging role of microRNAs in regulating the wasting response in individuals with chronic disease.


Asunto(s)
Fallo Renal Crónico/complicaciones , MicroARNs/fisiología , Diálisis Renal/efectos adversos , Síndrome Debilitante/etiología , Enfermedad Crónica , Humanos , Músculo Esquelético/patología
8.
J Acoust Soc Am ; 147(3): 1709, 2020 03.
Artículo en Inglés | MEDLINE | ID: mdl-32237844

RESUMEN

Behavioral guidance systems are commonly used in freshwater fish conservation. The biological relevance of sound to fish and recorded responses to human-generated noise supports the viability of the use of acoustics as an effective stimulus in such technologies. Relatively little information exists on the long-term responses and recovery of fish to repeated acoustic exposures. In a controlled laboratory study, the response and tolerance of Eurasian minnow (Phoxinus phoxinus) shoals to tonal signals (150 Hz of 1 s pulse duration) differing only in temporal characteristics ("continuous," "slow," "intermediate," or "fast" pulse repetition rate) were investigated. In comparison to independent control groups, fish increased their mean group swimming speed, decreased inter-individual distance, and became more aligned in response to the onset of all four acoustic treatments. The magnitude of response, and time taken to develop a tolerance to a treatment differed according to pulse repetition rate. Groups were found to have the greatest and longest lasting response to tone sequences tested in this study when they were pulsed at an intermediate rate of 0.2 s-1. This study illustrates the importance of understanding the response of fish to acoustic signals, and will assist toward the development of longer-term effective acoustic guidance systems.


Asunto(s)
Cyprinidae , Acústica , Animales , Humanos , Ruido , Conducta Social , Sonido
9.
Mar Policy ; 120: 104114, 2020 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-32834407

RESUMEN

Brexit creates a systemic shock that provides a unique opportunity for the UK to implement a new sustainable Fisheries Policy to better manage the multiple stocks on which future fishers will depend on leaving the European Union. At the same time, the global slowdown of commercial fishing as a result of COVID-19 has reduced pressure on some threatened stocks to levels not seen since the Second World War. In combination, Brexit and the COVID-19 slowdown have created a unique opportunity to facilitate the recovery of a threatened resource. Nevertheless, challenges remain as fisheries represent only 0.12% of UK economic output, presenting a risk that opportunities for more sustainable management will be lost during wider trade negotiations. Reduced fishing pressure during the COVID-19 era will enable stocks an opportunity to recover if supported by a new UK Fisheries Policy that focuses on: (a) re-establishing the role of Maximum Sustainable Yield to set limits that enable the recovery of fish populations initiated during the COVID-19 era; (b) ensuring that catch targets are set with the aim to maintain biomass at 120% of that which will achieve Maximum Sustainable Yield; (c) improving coherent resource management that also considers the expensive use of carbon associated with unsustainable fishing, and the need to protect fish throughout their life-cycle; and (d) constructing and effectively enforcing protection of a resilient network of Marine Protected Areas despite potential protests from EU member states.

10.
Thorax ; 74(2): 164-176, 2019 02.
Artículo en Inglés | MEDLINE | ID: mdl-30554141

RESUMEN

INTRODUCTION: Skeletal muscle dysfunction is a clinically important complication of pulmonary arterial hypertension (PAH). Growth/differentiation factor 15 (GDF-15), a prognostic marker in PAH, has been associated with muscle loss in other conditions. We aimed to define the associations of GDF-15 and muscle wasting in PAH, to assess its utility as a biomarker of muscle loss and to investigate its downstream signalling pathway as a therapeutic target. METHODS: GDF-15 levels and measures of muscle size and strength were analysed in the monocrotaline (MCT) rat, Sugen/hypoxia mouse and in 30 patients with PAH. In C2C12 myotubes the downstream targets of GDF-15 were identified. The pathway elucidated was then antagonised in vivo. RESULTS: Circulating GDF-15 levels correlated with tibialis anterior (TA) muscle fibre diameter in the MCT rat (Pearson r=-0.61, p=0.003). In patients with PAH, plasma GDF-15 levels of <564 pg/L predicted those with preserved muscle strength with a sensitivity and specificity of ≥80%. In vitro GDF-15 stimulated an increase in phosphorylation of TGFß-activated kinase 1 (TAK1). Antagonising TAK1, with 5(Z)-7-oxozeaenol, in vitro and in vivo led to an increase in fibre diameter and a reduction in mRNA expression of atrogin-1 in both C2C12 cells and in the TA of animals who continued to grow. Circulating GDF-15 levels were also reduced in those animals which responded to treatment. CONCLUSIONS: Circulating GDF-15 is a biomarker of muscle loss in PAH that is responsive to treatment. TAK1 inhibition shows promise as a method by which muscle atrophy may be directly prevented in PAH. TRIAL REGISTRATION NUMBER: NCT01847716; Results.


Asunto(s)
Factor 15 de Diferenciación de Crecimiento/metabolismo , Hipertensión Pulmonar/complicaciones , Quinasas Quinasa Quinasa PAM/metabolismo , Atrofia Muscular/etiología , Factor de Crecimiento Transformador beta/metabolismo , Adulto , Animales , Biomarcadores/metabolismo , Western Blotting , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Hipertensión Pulmonar/metabolismo , Inmunohistoquímica , Masculino , Ratones , Persona de Mediana Edad , Músculo Esquelético/metabolismo , Atrofia Muscular/metabolismo , Ratas , Ratas Sprague-Dawley , Reacción en Cadena en Tiempo Real de la Polimerasa , Transducción de Señal
11.
J Fish Biol ; 95(1): 228-237, 2019 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-30251260

RESUMEN

According to Signal Detection Theory, the ability to detect a stimulus (discriminability, d') is inversely related to the magnitude of internal and external noise. In this study, downstream moving brown trout Salmo trutta were used to investigate whether external hydrodynamic noise (in this case turbulence) could mask a signal that induced an unwanted response, such as rejecting accelerating velocity gradients commonly encountered at entrances to fish bypass channels. S. trutta behaviour was quantified in the absence (control) or presence of an accelerating velocity gradient created by an unconstricted or constricted channel, respectively, under two levels (low and high) of background turbulent kinetic energy (hydrodynamic noise). Experiments were conducted in an indoor recirculating flume in the dark and a range of passage metrics were quantified. Under the control condition, most (ca. 91%) S. trutta passed, usually oriented downstream (67%), with minimal delay (median 0.13 min). In comparison, fewer S. trutta (ca. 43%) passed under constricted conditions, they tended to orient facing into the flow (ca. 64%) and delay was greater (median > 20 min). When viewed from a coarse-scale perspective, discriminability of the velocity gradient was lower when turbulence was high suggesting masking of the signal occurred. However, the resulting increase in the percentage of fish that passed, decrease in time to pass and reduction in the distance at which S. trutta reacted (switched orientation) was subtle and non-significant. Despite the mixed results obtained, the use of masking to manipulate an animal's perception of environmental stimuli as a fisheries management tool is conceptually valid and the results of this experiment present a useful stepping stone for future research.


Asunto(s)
Conducta Animal , Estimulación Física , Trucha/fisiología , Migración Animal , Animales , Explotaciones Pesqueras , Hidrodinámica , Ríos/química , Alimentos Marinos , Movimientos del Agua
12.
Chron Respir Dis ; 16: 1479973119843650, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31131626

RESUMEN

In chronic obstructive pulmonary disease (COPD), lower limb dysfunction is associated with reduced exercise capacity, increased hospitalizations and mortality. We investigated sex differences in the prevalence of quadriceps dysfunction and fibre abnormalities in a large COPD cohort, controlling for the normal sex differences in health. We compared existing data from 76 male and 38 female COPD patients where each variable was expressed as a function of gender-specific normal values (obtained from 16 male and 14 female controls). Female COPD patients had lower quadriceps muscle strength and peak workload on a maximal incremental cycle ergometry protocol compared to male patients. Female patients had a smaller type II fibre cross-sectional area (CSA) compared to male patients, suggesting a greater female preponderance to fibre atrophy, although this result was largely driven by a few male patients with a large type II fibre CSA. Female patients had significantly higher concentrations of a number of plasma pro-inflammatory cytokines including tumour necrosis factor alpha and interleukin 8 (IL8), but not lower levels of physical activity or arterial oxygenation, compared to males. Our data confirm results from a previous small study and suggest that female COPD patients have a greater prevalence of muscle wasting and weakness. Larger studies investigating sex differences in COPD-related muscle atrophy and weakness are needed, as the results will have implications for monitoring in clinical practice and for design of clinical trials evaluating novel muscle anabolic agents.


Asunto(s)
Fibras Musculares Esqueléticas/patología , Debilidad Muscular/fisiopatología , Atrofia Muscular/fisiopatología , Enfermedad Pulmonar Obstructiva Crónica/fisiopatología , Músculo Cuádriceps/fisiopatología , Anciano , Análisis de los Gases de la Sangre , Ergometría , Ejercicio Físico , Femenino , Volumen Espiratorio Forzado , Humanos , Inflamación/metabolismo , Interleucina-8/metabolismo , Masculino , Persona de Mediana Edad , Fibras Musculares de Contracción Rápida/patología , Fibras Musculares de Contracción Lenta/patología , Fuerza Muscular , Debilidad Muscular/etiología , Debilidad Muscular/metabolismo , Debilidad Muscular/patología , Músculo Esquelético/patología , Músculo Esquelético/fisiopatología , Atrofia Muscular/etiología , Atrofia Muscular/metabolismo , Atrofia Muscular/patología , FN-kappa B/metabolismo , Tamaño de los Órganos , Consumo de Oxígeno , Enfermedad Pulmonar Obstructiva Crónica/complicaciones , Enfermedad Pulmonar Obstructiva Crónica/metabolismo , Músculo Cuádriceps/patología , Receptores del Factor de Necrosis Tumoral/metabolismo , Factores Sexuales , Factor de Transcripción AP-1/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Prueba de Paso
13.
Pflugers Arch ; 470(9): 1359-1376, 2018 09.
Artículo en Inglés | MEDLINE | ID: mdl-29797067

RESUMEN

Kv7 channels determine the resting membrane potential of neurons and regulate their excitability. Even though dysfunction of Kv7 channels has been linked to several debilitating childhood neuronal disorders, the ontogeny of the constituent genes, which encode Kv7 channels (KNCQ), and expression of their subunits have been largely unexplored. Here, we show that developmentally regulated expression of specific KCNQ mRNA and Kv7 channel subunits in mouse and human striatum is crucial to the functional maturation of mouse striatal neurons and human-induced pluripotent stem cell-derived neurons. This demonstrates their pivotal role in normal development and maturation, the knowledge of which can now be harnessed to synchronise and accelerate neuronal differentiation of stem cell-derived neurons, enhancing their utility for disease modelling and drug discovery.


Asunto(s)
Células Madre Pluripotentes Inducidas/metabolismo , Canal de Potasio KCNQ1/metabolismo , Neuronas/metabolismo , Regulación hacia Arriba/fisiología , Animales , Diferenciación Celular/fisiología , Células Cultivadas , Humanos , Potenciales de la Membrana/fisiología , Ratones , ARN Mensajero/metabolismo
14.
Am J Respir Crit Care Med ; 196(11): 1422-1433, 2017 12 01.
Artículo en Inglés | MEDLINE | ID: mdl-28809518

RESUMEN

RATIONALE: Loss of skeletal muscle mass and function is a common consequence of critical illness and a range of chronic diseases, but the mechanisms by which this occurs are unclear. OBJECTIVES: To identify microRNAs (miRNAs) that were increased in the quadriceps of patients with muscle wasting and to determine the molecular pathways by which they contributed to muscle dysfunction. METHODS: miRNA-542-3p/5p (miR-542-3p/5p) were quantified in the quadriceps of patients with chronic obstructive pulmonary disease and intensive care unit-acquired weakness (ICUAW). The effect of miR-542-3p/5p was determined on mitochondrial function and transforming growth factor-ß signaling in vitro and in vivo. MEASUREMENTS AND MAIN RESULTS: miR-542-3p/5p were elevated in patients with chronic obstructive pulmonary disease but more markedly in patients with ICUAW. In vitro, miR-542-3p suppressed the expression of the mitochondrial ribosomal protein MRPS10 and reduced 12S ribosomal RNA (rRNA) expression, suggesting mitochondrial ribosomal stress. miR-542-5p increased nuclear phospho-SMAD2/3 and suppressed expression of SMAD7, SMURF1, and PPP2CA, proteins that inhibit or reduce SMAD2/3 phosphorylation, suggesting that miR-542-5p increased transforming growth factor-ß signaling. In mice, miR-542 overexpression caused muscle wasting, and reduced mitochondrial function, 12S rRNA expression, and SMAD7 expression, consistent with the effects of the miRNAs in vitro. Similarly, in patients with ICUAW, the expression of 12S rRNA and of the inhibitors of SMAD2/3 phosphorylation were reduced, indicative of mitochondrial ribosomal stress and increased transforming growth factor-ß signaling. In patients undergoing aortic surgery, preoperative levels of miR-542-3p/5p were positively correlated with muscle loss after surgery. CONCLUSIONS: Elevated miR-542-3p/5p may cause muscle atrophy in intensive care unit patients through the promotion of mitochondrial dysfunction and activation of SMAD2/3 phosphorylation.


Asunto(s)
Cuidados Críticos , MicroARNs/metabolismo , Mitocondrias/metabolismo , Debilidad Muscular/metabolismo , Músculo Cuádriceps/metabolismo , Proteínas Smad/metabolismo , Animales , Modelos Animales de Enfermedad , Humanos , Unidades de Cuidados Intensivos , Masculino , Ratones , Transducción de Señal , Factor de Crecimiento Transformador beta/metabolismo
15.
Muscle Nerve ; 55(6): 902-912, 2017 06.
Artículo en Inglés | MEDLINE | ID: mdl-27680944

RESUMEN

INTRODUCTION: Quadriceps dysfunction is important in chronic obstructive pulmonary disease (COPD), with an associated increased proportion of type II fibers. Investigation of protein synthesis and degradation has yielded conflicting results, possibly due to study of whole biopsy samples, whereas signaling may be fiber-specific. Our objective was to develop a method for fiber-specific gene expression analysis. METHODS: 12 COPD and 6 healthy subjects underwent quadriceps biopsy. Cryosections were immunostained for type II fibers, which were separated using laser capture microdissection (LCM). Whole muscle and different fiber populations were subject to quantitative polymerase chain reaction. RESULTS: Levels of muscle-RING-finger-protein-1 and Atrogin-1 were lower in type II fibers of COPD versus healthy subjects (P = 0.02 and P = 0.03, respectively), but differences were not apparent in whole muscle or type I fibers. CONCLUSIONS: We describe a novel method for studying fiber-specific gene expression in optimum cutting temperature compound-embedded muscle specimens. LCM offers a more sensitive way to identify molecular changes in COPD muscle. Muscle Nerve 55: 902-912, 2017.


Asunto(s)
Regulación de la Expresión Génica/fisiología , Captura por Microdisección con Láser , Fibras Musculares Esqueléticas/metabolismo , Enfermedad Pulmonar Obstructiva Crónica/patología , Transducción de Señal/fisiología , Adulto , Anciano , Anciano de 80 o más Años , Estudios de Casos y Controles , Humanos , Péptidos y Proteínas de Señalización Intracelular/genética , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Masculino , Persona de Mediana Edad , Fibras Musculares Esqueléticas/clasificación , Fibras Musculares Esqueléticas/patología , Proteínas Musculares/genética , Proteínas Musculares/metabolismo , Músculo Esquelético/fisiopatología , Cadenas Pesadas de Miosina/metabolismo , Fosfotransferasas (Aceptor de Grupo Alcohol)/genética , Fosfotransferasas (Aceptor de Grupo Alcohol)/metabolismo , Proyectos Piloto , Proteínas Ligasas SKP Cullina F-box/genética , Proteínas Ligasas SKP Cullina F-box/metabolismo , Estadísticas no Paramétricas
16.
Am J Physiol Cell Physiol ; 310(7): C520-41, 2016 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-26718628

RESUMEN

Although numerous protocols have been developed for differentiation of neurons from a variety of pluripotent stem cells, most have concentrated on being able to specify effectively appropriate neuronal subtypes and few have been designed to enhance or accelerate functional maturity. Of those that have, most employ time courses of functional maturation that are rather protracted, and none have fully characterized all aspects of neuronal function, from spontaneous action potential generation through to postsynaptic receptor maturation. Here, we describe a simple protocol that employs the sequential addition of just two supplemented media that have been formulated to separate the two key phases of neural differentiation, the neurogenesis and synaptogenesis, each characterized by different signaling requirements. Employing these media, this new protocol synchronized neurogenesis and enhanced the rate of maturation of pluripotent stem cell-derived neural precursors. Neurons differentiated using this protocol exhibited large cell capacitance with relatively hyperpolarized resting membrane potentials; moreover, they exhibited augmented: 1) spontaneous electrical activity; 2) regenerative induced action potential train activity; 3) Na(+) current availability, and 4) synaptic currents. This was accomplished by rapid and uniform development of a mature, inhibitory GABAAreceptor phenotype that was demonstrated by Ca(2+) imaging and the ability of GABAAreceptor blockers to evoke seizurogenic network activity in multielectrode array recordings. Furthermore, since this protocol can exploit expanded and frozen prepatterned neural progenitors to deliver mature neurons within 21 days, it is both scalable and transferable to high-throughput platforms for the use in functional screens.


Asunto(s)
Técnicas de Cultivo de Célula/métodos , Diferenciación Celular/fisiología , Medios de Cultivo/química , Células Madre Pluripotentes Inducidas/citología , Células-Madre Neurales/citología , Western Blotting , Ciclo Celular/fisiología , Línea Celular , Técnicas de Cocultivo , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Glucógeno Sintasa Quinasa 3/metabolismo , Glucógeno Sintasa Quinasa 3 beta , Humanos , Procesamiento de Imagen Asistido por Computador , Inmunohistoquímica , Células Madre Pluripotentes Inducidas/metabolismo , Microscopía Electrónica de Rastreo , Células-Madre Neurales/metabolismo , Neurogénesis/fisiología , Técnicas de Placa-Clamp , Receptores de GABA-A/metabolismo
17.
J Physiol ; 594(22): 6583-6594, 2016 11 15.
Artículo en Inglés | MEDLINE | ID: mdl-27616476

RESUMEN

Neurons differentiated from pluripotent stem cells using established neural culture conditions often exhibit functional deficits. Recently, we have developed enhanced media which both synchronize the neurogenesis of pluripotent stem cell-derived neural progenitors and accelerate their functional maturation; together these media are termed SynaptoJuice. This pair of media are pro-synaptogenic and generate authentic, mature synaptic networks of connected forebrain neurons from a variety of induced pluripotent and embryonic stem cell lines. Such enhanced rate and extent of synchronized maturation of pluripotent stem cell-derived neural progenitor cells generates neurons which are characterized by a relatively hyperpolarized resting membrane potential, higher spontaneous and induced action potential activity, enhanced synaptic activity, more complete development of a mature inhibitory GABAA receptor phenotype and faster production of electrical network activity when compared to standard differentiation media. This entire process - from pre-patterned neural progenitor to active neuron - takes 3 weeks or less, making it an ideal platform for drug discovery and disease modelling in the fields of human neurodegenerative and neuropsychiatric disorders, such as Huntington's disease, Parkinson's disease, Alzheimer's disease and Schizophrenia.


Asunto(s)
Calcio/metabolismo , Diferenciación Celular/fisiología , Neuronas/metabolismo , Neuronas/fisiología , Células Madre Pluripotentes/metabolismo , Células Madre Pluripotentes/fisiología , Receptores de GABA-A/metabolismo , Animales , Humanos , Neurogénesis/fisiología
18.
J Cell Physiol ; 231(6): 1375-84, 2016 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-26580832

RESUMEN

Voltage-gated potassium Kv1.2 channels play pivotal role in maintaining of resting membrane potential and, consequently, regulation of cellular excitability of neurons. Endogenously generated electric field (EF) have been proven as an important regulator for cell migration and tissue repair. The mechanisms of ion channel involvement in EF-induced cell responses are extensively studied but largely are poorly understood. In this study we generated three COS-7 clones with different expression levels of Kv1.2 channel, and confirmed their functional variations with patch clamp analysis. Time-lapse imaging analysis showed that EF-induced cell migration response was Kv1.2 channel expression level depended. Inhibition of Kv1.2 channels with charybdotoxin (ChTX) constrained the sensitivity of COS-7 cells to EF stimulation more than their motility. Immunocytochemistry and pull-down analyses demonstrated association of Kv1.2 channels with actin-binding protein cortactin and its re-localization to the cathode-facing membrane at EF stimulation, which confirms the mechanism of EF-induced directional migration. This study displays that Kv1.2 channels represent an important physiological link in EF-induced cell migration. The described mechanism suggests a potential application of EF which may improve therapeutic performance in curing injuries of neuronal and/or cardiac tissue repair, post operational therapy, and various degenerative syndromes.


Asunto(s)
Movimiento Celular , Canal de Potasio Kv.1.1/metabolismo , Animales , Células COS , Movimiento Celular/efectos de los fármacos , Chlorocebus aethiops , Cortactina/metabolismo , Relación Dosis-Respuesta a Droga , Estimulación Eléctrica , Inmunoprecipitación , Canal de Potasio Kv.1.1/antagonistas & inhibidores , Canal de Potasio Kv.1.1/genética , Potenciales de la Membrana , Técnicas de Placa-Clamp , Bloqueadores de los Canales de Potasio/farmacología , Unión Proteica , Transducción de Señal , Factores de Tiempo , Imagen de Lapso de Tiempo , Transfección
19.
J Exp Biol ; 219(Pt 21): 3480-3491, 2016 11 01.
Artículo en Inglés | MEDLINE | ID: mdl-27591311

RESUMEN

It is commonly assumed that stream-dwelling fish should select positions where they can reduce energetic costs relative to benefits gained and enhance fitness. However, the selection of appropriate hydrodynamic metrics that predict space use is the subject of recent debate and a cause of controversy. This is for three reasons: (1) flow characteristics are often oversimplified, (2) confounding variables are not always controlled and (3) there is limited understanding of the explanatory mechanisms that underpin the biophysical interactions between fish and their hydrodynamic environment. This study investigated the space use of brown trout, Salmo trutta, in a complex hydrodynamic flow field created using an array of different sized vertically oriented cylinders in a large open-channel flume in which confounding variables were controlled. A hydrodynamic drag function (D) based on single-point time-averaged velocity statistics that incorporates the influence of turbulent fluctuations was used to infer the energetic cost of steady swimming. Novel hydrodynamic preference curves were developed and used to assess the appropriateness of D as a descriptor of space use compared with other commonly used metrics. Zones in which performance-enhancing swimming behaviours (e.g. Kármán gaiting, entraining and bow riding) that enable fish to hold position while reducing energetic costs (termed 'specialised behaviours') were identified and occupancy was recorded. We demonstrate that energy conservation strategies play a key role in space use in an energetically taxing environment with the majority of trout groups choosing to frequently occupy areas in which specialised behaviours may be adopted or by selecting low-drag regions.


Asunto(s)
Ambiente , Hidrodinámica , Reología , Trucha/fisiología , Animales , Conducta Animal/fisiología
20.
Adv Exp Med Biol ; 875: 747-53, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-26611028

RESUMEN

Shipping creates large near-field background noises at levels similar to or higher than fish vocalizations and in the same critical bandwidths. This noise has the potential to "mask" biologically important signals and prevent fish from hearing them; any interference with the detection and recognition of sounds may impact fish survival. The Lombard effect, whereby vocalizations are altered to reduce or exclude masking effects, is an adaptation that has been observed in mammals and birds. Research is needed to establish whether the Lombard effect occurs in fish to gain a better understanding of the implications of noise pollution on fish populations.


Asunto(s)
Peces/fisiología , Ruido , Enmascaramiento Perceptual/fisiología , Navíos , Vocalización Animal/fisiología , Animales
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