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BACKGROUND: This study aimed to investigate the effectiveness of a 6-month home-based high-intensity interval training (HIIT) intervention to improve peak oxygen consumption (VÌO2peak) and lactate threshold (LT) in older adults. METHODS: Two hundred thirty-three healthy older adults (60-84 years; 54% females) were randomly assigned to either 6-month, thrice-weekly home-based HIIT (once-weekly circuit training and twice-weekly interval training) or a passive control group. Exercise sessions were monitored using a Polar watch and a logbook for objective and subjective data, respectively, and guided by a personal coach. The outcomes were assessed using a modified Balke protocol combining VÌO2peak and LT measures. General linear regression models assessed between-group differences in change and within-group changes for each outcome. RESULTS: There was a significant between-group difference in the pre-to-post change in VÌO2peak (difference: 1.8 [1.2; 2.3] mL/kg/min; exercise: +1.4 [1.0; 1.7] mL/kg/min [~5%]; control: -0.4 [-0.8; -0.0] mL/kg/min [approximately -1.5%]; effect size [ES]: 0.35). Compared with controls, the exercise group had lower blood lactate concentration (-0.7 [-0.9; -0.4] mmol/L, ES: 0.61), % of peak heart rate (-4.4 [-5.7; -3.0], ES: 0.64), and % of VÌO2peak (-4.5 [-6.1; -2.9], ES: 0.60) at the intensity corresponding to preintervention LT and achieved a higher treadmill stage (% incline) at LT (0.6 [0.3; 0.8]; ES: 0.47), following the intervention. CONCLUSION: This study highlights the effectiveness of a home-based HIIT intervention as an accessible and equipment-minimal strategy to induce clinically meaningful improvements in cardiorespiratory fitness in older adults. Over 6 months, the exercise group showed larger improvements in all outcomes compared with the control group. Notably, the LT outcome exhibited a more pronounced magnitude of change than VÌO2peak.
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Aptidão Cardiorrespiratória , Treinamento Intervalado de Alta Intensidade , Ácido Láctico , Consumo de Oxigênio , Humanos , Feminino , Aptidão Cardiorrespiratória/fisiologia , Masculino , Idoso , Consumo de Oxigênio/fisiologia , Treinamento Intervalado de Alta Intensidade/métodos , Pessoa de Meia-Idade , Idoso de 80 Anos ou mais , Ácido Láctico/sangue , Frequência Cardíaca/fisiologiaRESUMO
BACKGROUND: It is well-established that cross-sectional measurements of poor body composition are associated with impaired physical function and that power training effectively enhances total lean mass and physical function in older adults. However, it is unclear if power training-induced changes in body composition are associated with improved physical function in older adults. AIM: The present study investigated associations between body composition and physical function cross-sectionally and with power training-induced changes in older men. METHODS: Forty-nine older men (68 ± 5 yrs) completed a 10-week biweekly power training intervention. Body composition was measured using dual-energy X-ray absorptiometry. Physical function was assessed as a composite Z-score combining measures from Sit-to-stand power, Timed up-and-go time, and loaded and unloaded Stair-climbing time (15 steps). Linear and quadratic regression analyses were performed to assess associations between body composition and physical function. RESULTS: At baseline, total (R2 = 0.11, p < 0.05) and percentage body fat (R2 = 0.15, p < 0.05) showed a non-linear relationship with physical function. The apex of the quadratic regression for body composition was 21.5% body fat. Furthermore, there was a non-linear relationship between changes in body fat percentage and physical function from pre- to post-intervention (R2 = 0.15, p < 0.05). CONCLUSION: The present study's findings indicate that participants with a body composition of ~20% body fat displayed the highest level of physical function at baseline. Furthermore, despite small pre-post changes in body fat, the results indicate that those who either preserved their body fat percentage or experienced minor alterations observed the greatest improvements in physical function.
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Composição Corporal , Força Muscular , Masculino , Humanos , Idoso , Estudos Transversais , Tecido AdiposoRESUMO
There has recently been great interest in the use of accelerometers onboard electronic transmitters to characterise various aspects of the ecology of wild animals. We review use cases and outline how these tools can provide opportunities for studying activity and survival, exercise physiology of wild animals, the response to stressors, energy landscapes and conservation planning tools, and the means with which to identify behaviours remotely from transmitted data. Accelerometer transmitters typically send data summaries to receivers at fixed intervals after filtering out static acceleration and calculating root-mean square error or overall dynamic body action of 2- or 3-axis acceleration values (often at 5-12.5 Hz) from dynamic acceleration onboard the tag. Despite the popularity of these transmitters among aquatic ecologists, we note that there is wide variation in the sampling frequencies and windows used among studies that will potentially affect the ability to make comparisons in the future. Accelerometer transmitters will likely become increasingly popular tools for studying finer scale details about cryptic species that are difficult to recapture and hence not suitable for studies using data loggers. We anticipate that there will continue to be opportunities to adopt methods used for analysing data from loggers to datasets generated from acceleration transmitters, to generate new knowledge about the ecology of aquatic animals.
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AIMS: Assessment of left ventricular (LV) function by echocardiography is hampered by modest test-retest reproducibility. A novel artificial intelligence (AI) method based on deep learning provides fully automated measurements of LV global longitudinal strain (GLS) and may improve the clinical utility of echocardiography by reducing user-related variability. The aim of this study was to assess within-patient test-retest reproducibility of LV GLS measured by the novel AI method in repeated echocardiograms recorded by different echocardiographers and to compare the results to manual measurements. METHODS: Two test-retest data sets (n = 40 and n = 32) were obtained at separate centers. Repeated recordings were acquired in immediate succession by 2 different echocardiographers at each center. For each data set, 4 readers measured GLS in both recordings using a semiautomatic method to construct test-retest interreader and intrareader scenarios. Agreement, mean absolute difference, and minimal detectable change (MDC) were compared to analyses by AI. In a subset of 10 patients, beat-to-beat variability in 3 cardiac cycles was assessed by 2 readers and AI. RESULTS: Test-retest variability was lower with AI compared with interreader scenarios (data set I: MDC = 3.7 vs 5.5, mean absolute difference = 1.4 vs 2.1, respectively; data set II: MDC = 3.9 vs 5.2, mean absolute difference = 1.6 vs 1.9, respectively; all P < .05). There was bias in GLS measurements in 13 of 24 test-retest interreader scenarios (largest bias, 3.2 strain units). In contrast, there was no bias in measurements by AI. Beat-to-beat MDCs were 1,5, 2.1, and 2.3 for AI and the 2 readers, respectively. Processing time for analyses of GLS by the AI method was 7.9 ± 2.8 seconds. CONCLUSION: A fast AI method for automated measurements of LV GLS reduced test-retest variability and removed bias between readers in both test-retest data sets. By improving the precision and reproducibility, AI may increase the clinical utility of echocardiography.
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Aprendizado Profundo , Disfunção Ventricular Esquerda , Humanos , Reprodutibilidade dos Testes , Inteligência Artificial , Função Ventricular Esquerda , Ecocardiografia/métodos , Disfunção Ventricular Esquerda/diagnóstico por imagem , Volume SistólicoRESUMO
Measurements of cardiac function such as left ventricular ejection fraction and myocardial strain are typically based on 2-D ultrasound imaging. The reliability of these measurements depends on the correct pose of the transducer such that the 2-D imaging plane properly aligns with the heart for standard measurement views and is thus dependent on the operator's skills. We propose a deep learning tool that suggests transducer movements to help users navigate toward the required standard views while scanning. The tool can simplify echocardiography for less experienced users and improve image standardization for more experienced users. Training data were generated by slicing 3-D ultrasound volumes, which permits simulation of the movements of a 2-D transducer. Neural networks were further trained to calculate the transducer position in a regression fashion. The method was validated and tested on 2-D images from several data sets representative of a prospective clinical setting. The method proposed the adequate transducer movement 75% of the time when averaging over all degrees of freedom and 95% of the time when considering transducer rotation solely. Real-time application examples illustrate the direct relation between the transducer movements, the ultrasound image and the provided feedback.
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Ecocardiografia Tridimensional , Função Ventricular Esquerda , Volume Sistólico , Reprodutibilidade dos Testes , Estudos Prospectivos , Ecocardiografia/métodosRESUMO
The study aimed to investigate the effectiveness of an individualized power training program based on force-velocity (FV) profiling on physical function, muscle morphology, and neuromuscular adaptations in older men. Forty-nine healthy men (68 ± 5 years) completed a 10-week training period to enhance muscular power. They were randomized to either a generic power training group (GPT) or an individualized power training group (IPT). Unlike generic training, individualized training was based on low- or high-resistance exercises, from an initial force-velocity profile. Lower-limb FV profile was measured in a pneumatic leg-press, and physical function was assessed as timed up-and-go time (TUG), sit-to-stand power, grip strength, and stair-climbing time (loaded [20kg] and unloaded). Vastus lateralis morphology was measured with ultrasonography. Rate of force development (RFD) and rate of myoelectric activity (RMA) were measured during an isometric knee extension. The GPT group improved loaded stair-climbing time (6.3 ± 3.8 vs. 2.3% ± 7.3%, p = 0.04) more than IPT. Both groups improved stair-climbing time, sit to stand, and leg press power, grip strength, muscle thickness, pennation angle, fascicle length, and RMA from baseline (p < 0.05). Only GPT increased loaded stair-climbing time and RFD (p < 0.05). An individualized power training program based on FV profiling did not improve physical function to a greater degree than generic power training. A generic power training approach combining both heavy and low loads might be advantageous through eliciting both force- and velocity-related neuromuscular adaptions with a concomitant increase in muscular power and physical function in older men.
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Força Muscular , Treinamento Resistido , Adaptação Fisiológica , Idoso , Teste de Esforço , Humanos , Masculino , Força Muscular/fisiologia , Músculo Esquelético/fisiologia , Músculo Quadríceps/diagnóstico por imagemRESUMO
Salmonids are some of the most widely studied species of fish worldwide. They span freshwater rivers and lakes to fjords and oceans; they include short- and long-distance anadromous migrants, as well as partially migratory and non-migratory populations; and exhibit both semelparous and iteroparous reproduction. Salmonid life-history strategies represent some of the most diverse on the planet. For this reason, salmonids provide an especially interesting model to study the drivers of these different life-history pathways. Over the past few decades, numerous studies and reviews have been published, although most have focused on ultimate considerations where expected reproductive success of different developmental or life-history strategies are compared. Those that considered proximate causes generally focused on genetics or the environment, with less consideration of physiology. Our objective was therefore to review the existing literature on the role of physiology as a proximate driver for life-history strategies in salmonids. This link is necessary to explore since physiology is at the core of biological processes influencing energy acquisition and allocation. Energy acquisition and allocation processes, in turn, can affect life histories. We find that life-history strategies are driven by a range of physiological processes, ranging from metabolism and nutritional status to endocrinology. Our review revealed that the role of these physiological processes can vary across species and individuals depending on the life-history decision(s) to be made. In addition, while findings sometimes vary by species, results appear to be consistent in species with similar life cycles. We conclude that despite much work having been conducted on the topic, the study of physiology and its role in determining life-history strategies in salmonids remains somewhat unexplored, particularly for char and trout (excluding brown trout) species. Understanding these mechanistic links is necessary if we are to understand adequately how changing environments will impact salmonid populations.
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Salmonidae , Migração Animal , Animais , Humanos , Lagos , Rios , TrutaRESUMO
Subsequent to their introduction in the 1950s, Arctic charr Salvelinus alpinus have been able to establish a self-sustaining population that has adapted to the unique conditions of the sub-Antarctic Kerguelen Islands. Here, 48 individuals (198-415 mm) were caught with gillnets and their basic biology and feeding ecology were examined using stable isotope analysis. The Lac des Fougères population split use of littoral and pelagic resources evenly, although larger fish relied more heavily on littoral production and appear to follow the size-dependent life history habitat template seen in many Scandinavian lakes where smaller sized individuals occupy the pelagic zone and larger individuals dominate the littoral habitat. In Kerguelen, Arctic charr mature at the same ages (5.6 years) as Arctic charr in both sub-Arctic and Arctic lakes. Although mortality was average in comparison to comparator sub-Arctic lakes, it was high in comparison to Arctic lakes. Maximal age (>7+) was at the lower end of the range typically seen in sub-Arctic lakes. Although they inhabit a resource-poor environment, Kerguelen Arctic charr showed no evidence of cannibalism. Thus, while Arctic charr can survive and reproduce in the relatively unproductive Kerguelen lake environments, survival and growth nevertheless appear to be traded off against survival and longevity. The uniqueness of the population location and the recency of its introduction suggest that further monitoring of the population has the potential to yield valuable insights into both the adaptability of the species and its likely responses to ongoing large-scale environmental change as represented by climate change.
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Comportamento Alimentar/fisiologia , Reprodução/fisiologia , Truta/fisiologia , Animais , Regiões Antárticas , Tamanho Corporal , Mudança Climática , Ecossistema , Ilhas , LagosRESUMO
Wild fish are confronting changing pathogen dynamics arising from anthropogenic disturbance and climate change. Pathogens can influence animal behaviour and life histories, yet there are little such data from fish in the high north where pathogen dynamics may differ from comparatively southern regions. We aimed to compare the pathogen communities of 160 wild anadromous brown trout in two fjords in northern Norway and to determine whether pathogens influenced area use or return to spawn. Application of high-throughput qPCR detected 11 of the 46 pathogens screened for; most frequently encountered were Ichthyobodo spp., Flavobacterium psychrophilum and Candidatus Branchiomonas cysticola. The rate of returning to freshwater during the spawning season was significantly lower for the Skjerstadfjord fish. Piscichlamydia salmonis and F. psychrophilum were indicator species for the Skjerstadfjord and pathogen communities in the two fjords differed according to perMANOVA. Individual length, Fulton's condition factor and the time between first and last detection of the fish were not related to the presence of pathogens ordinated using non-metric multidimensional scaling (NMDS). However, there was evidence that pathogen load was correlated with the expression of smoltification genes, which are upregulated by salmonids in freshwater. Correspondingly, percentage of time in freshwater after release was longer for fish with greater pathogen burdens.
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Migração Animal , Ecossistema , Doenças dos Peixes/microbiologia , Truta , Animais , Carga Bacteriana , Doenças dos Peixes/parasitologia , Doenças dos Peixes/virologia , Sequenciamento de Nucleotídeos em Larga Escala/veterinária , Noruega , Reação em Cadeia da Polimerase/veterinária , Carga ViralRESUMO
BACKGROUND: The use of assisted reproductive technology is increasing worldwide and conception after assisted reproduction currently comprises 3%-6% of birth cohorts in the Nordic countries. The risk of placenta-mediated pregnancy complications is greater after assisted reproductive technology compared with spontaneously conceived pregnancies. Whether the excess risk of placenta-mediated pregnancy complications in pregnancies following assisted reproduction has changed over time, is unknown. OBJECTIVES: To investigate whether time trends in risk of pregnancy complications (hypertensive disorders in pregnancy, placental abruption and placenta previa) differ for pregnancies after assisted reproductive technology compared with spontaneously conceived pregnancies during 3 decades of assisted reproduction treatment in the Nordic countries. STUDY DESIGN: In a population-based cohort study, with data from national health registries in Denmark (1994-2014), Finland (1990-2014), Norway (1988-2015) and Sweden (1988-2015), we included 6,830,578 pregnancies resulting in delivery. Among these, 146,998 (2.2%) were pregnancies after assisted reproduction (125,708 singleton pregnancies, 20,668 twin pregnancies and 622 of higher order plurality) and 6,683,132 (97.8%) pregnancies were conceived spontaneously (6,595,185 singleton pregnancies, 87,106 twin pregnancies and 1,289 of higher order plurality). We used logistic regression with post-estimation to estimate absolute risks and risk differences for each complication. We repeated analyses for singleton and twin pregnancies, separately. In subsamples with available information, we also adjusted for maternal body mass index, smoking during pregnancy, previous cesarean delivery, culture duration, and cryopreservation. RESULTS: The risk of each placental complication was consistently greater in pregnancies following assisted reproductive technology compared with spontaneously conceived pregnancies across the study period, except for hypertensive disorders in twin pregnancies, where risks were similar. Risk of hypertensive disorders increased over time in twin pregnancies for both conception methods, but more strongly for pregnancies following assisted reproductive technology (risk difference, 1.73 percentage points per 5 years; 95% confidence interval, 1.35-2.11) than for spontaneously conceived twins (risk difference, 0.75 percentage points; 95% confidence interval, 0.61-0.89). No clear time trends were found for hypertensive disorders in singleton pregnancies. Risk of placental abruption decreased over time in all groups. Risk differences were -0.16 percentage points (95% confidence interval, -0.19 to -0.12) and -0.06 percentage points (95% confidence interval, -0.06 to -0.05) for pregnancies after assisted reproduction and spontaneously conceived pregnancies, respectively, for singletons and multiple pregnancies combined. Over time, the risk of placenta previa increased in pregnancies after assisted reproduction among both singletons (risk difference, 0.21 percentage points; 95% confidence interval, 0.14-0.27) and twins (risk difference, 0.30 percentage points; 95% confidence interval, 0.16-0.43), but remained stable in spontaneously conceived pregnancies. When adjusting for culture duration, the temporal increase in placenta previa became weaker in all groups of assisted reproductive technology pregnancies, whereas adjustment for cryopreservation moderately attenuated trends in assisted reproductive technology twin pregnancies. CONCLUSIONS: The risk of placenta-mediated pregnancy complications following assisted reproductive technology remains higher compared to spontaneously conceived pregnancies, despite declining rates of multiple pregnancies. For hypertensive disorders in pregnancy and placental abruption, pregnancies after assisted reproduction follow the same time trends as the background population, whereas for placenta previa, risk has increased over time in pregnancies after assisted reproductive technology.
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Descolamento Prematuro da Placenta/epidemiologia , Diabetes Gestacional/epidemiologia , Placenta Prévia/epidemiologia , Complicações na Gravidez/epidemiologia , Técnicas de Reprodução Assistida/efeitos adversos , Descolamento Prematuro da Placenta/etiologia , Adulto , Fatores Etários , Diabetes Gestacional/etiologia , Feminino , Humanos , Incidência , Placenta Prévia/etiologia , Gravidez , Complicações na Gravidez/etiologia , Sistema de Registros , Risco , Países Escandinavos e Nórdicos , Adulto JovemRESUMO
Acellular polymer-calcium phosphate composites are promising bone graft materials. Hydrogels are suitable for providing a temporary matrix, while calcium phosphate minerals serve as ion depots for calcium and phosphate required for de novo bone formation. Crystalline calcium phosphates are stable under biological conditions and are commonly used in such scaffolds. However, the low solubility of these phases reduces the availability of free ions and potentially obstructs the remodelling necessary for the formation of mineralised tissue. Here, we investigate two different strategies to stabilise amorphous calcium phosphates in a synthetic polyethylene glycol-based hydrogel matrix. In vitro experiments mimicking an injectable application showed that amorphous calcium phosphate (ACP) of variable stability was formed in the hydrogel matrices. In additive-free composites, ACP transformed into brushite within minutes. Citrate or zinc additives were found to stabilise the formed ACP phase to different degrees. In the presence of citrate, ACP was stable for at least 2â¯h before it transformed into hydroxyapatite within 3-20â¯days. Partial calcium substitution with zinc (Zn/Caâ¯=â¯10%) produced zinc-doped ACP of high stability that did not show signs of crystallisation for at least 20â¯days. The presented methods and findings open new possibilities for the design of novel injectable synthetic bone graft materials. The possibility to produce ACP with tailorable stability promises great potential for creating temporary scaffolds with good osteogenic properties. STATEMENT OF SIGNIFICANCE: Synthetic hydrogel-calcium phosphate (CaP) composites are promising biomaterials to replace human- and animal-derived bone scaffolds. Most reported hydrogel-CaP composite materials employ crystalline CaP phases that lack the osteoinductive properties of autograft. Stabilising amorphous calcium phosphates (ACP) could overcome this limitation, readily delivering calcium and phosphate ions and facilitating remodelling into new bone tissue. The design of synthetic hydrogel-ACP scaffolds, however, requires more understanding of the mineralisation processes in such matrices. This study presents a model system to characterise the complex mineral formation and transformation processes within a hydrogel matrix. We demonstrate a facile route to produce self-mineralising injectable synthetic hydrogels and prove two different strategies to stabilise ACP for different periods within the formed composites.
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Fosfatos de Cálcio/química , Hidrogéis/química , Polietilenoglicóis/química , Animais , Fosfatos de Cálcio/farmacologia , Durapatita/química , Humanos , Hidrogéis/farmacologia , OsteogêneseRESUMO
Pancreas disease (PD) is a viral disease caused by Salmonid alphavirus (SAV) that affects farmed Atlantic salmon (Salmo salar L.) and rainbow trout (Oncorhynchus mykiss (Walbaum)) in the seawater phase. Since its first description in Scotland in 1976, a large number of studies have been conducted relating to the disease itself and to factors contributing to agent spread and disease occurrence. This paper summarizes the currently available, scientific information on the epidemiology of PD and its associated mitigation and control measures. Available literature shows infected farmed salmonids to be the main reservoir of SAV. Transmission between seawater sites occurs mainly passively by water currents or actively through human activity coupled with inadequate biosecurity measures. All available information suggests that the current fallowing procedures are adequate to prevent agent survival within the environment through the fallowing period and thus that a repeated disease outbreak at the same site is due to a new agent introduction. There has been no scientific evaluation of currently used on-site biosecurity measures, and there is limited information on the impact of available mitigation measures and control strategies.
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Infecções por Alphavirus/veterinária , Alphavirus/fisiologia , Doenças dos Peixes/epidemiologia , Oncorhynchus mykiss , Pancreatopatias/veterinária , Salmo salar , Infecções por Alphavirus/epidemiologia , Infecções por Alphavirus/virologia , Animais , Aquicultura , Europa (Continente)/epidemiologia , Doenças dos Peixes/virologia , Pancreatopatias/epidemiologia , Pancreatopatias/virologia , PrevalênciaAssuntos
Infecções por Alphavirus/veterinária , Alphavirus , Doenças dos Peixes/virologia , Perciformes , Infecções por Alphavirus/transmissão , Animais , Suscetibilidade a Doenças/veterinária , Doenças dos Peixes/transmissão , Pancreatopatias/veterinária , Pancreatopatias/virologia , Salmão/virologiaRESUMO
UNLABELLED: The modification of soft hydrogels with hard inorganic components is a method used to form composite materials with application in non-load-bearing bone tissue engineering. The inclusion of an inorganic component may provide mechanical enhancement, introduce osteoconductive or osteoinductive properties, or change other aspects of interactions between native or implanted cells and the material. A thorough understanding of the interactions between such components is needed to improve the rational design of such biomaterials. To achieve this goal, model systems which could allow study of the formation and transformation of mineral phases within a hydrogel network with a range of experimental methods and high spatial and time resolution are needed. Here, we report a detailed investigation of the formation and transformation process of calcium phosphate mineral within an alginate hydrogel matrix. A combination of optical microscopy, confocal Raman microspectroscopy and electron microscopy was used to investigate the spatial distribution, morphology and crystal phase of the calcium phosphate mineral, as well as to study transformation of the mineral phases during the hydrogel mineralization process and upon incubation in a simulated body fluid. It was found, that under the conditions used in this work, mineral initially formed as a metastable amorphous calcium phosphate phase (ACP). The ACP particles had a distinctive spherical morphology and transformed within minutes into brushite in the presence of brushite seed crystals or into octacalcium phosphate, when no seeds were present in the hydrogel matrix. Incubation of brushite-alginate composites in simulated body fluid resulted in formation of hydroxyapatite. The characterization strategy presented here allows for non-destructive, in situ observation of mineralization processes in optically transparent hydrogels with little to no sample preparation. STATEMENT OF SIGNIFICANCE: The precipitation and transformations of calcium phosphates (CaP) is a complex process, where both formation kinetics and the stability of different mineral phases control the outcome. This situation is even more complex if CaP is precipitated in a hydrogel matrix, where one can expect the organic matrix to modulate crystallization by introducing supersaturation gradients or changing the nucleation and growth kinetics of crystals. In this study we apply a range of characterization techniques to study the mineral formation and transformations of CaP within an alginate matrix with spatiotemporal resolution. It demonstrates how a detailed investigation of the mineral precipitation and transformations can aid in the future rational design of hydrogel-based materials for bone tissue engineering and studies of biomineralization processes.
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Alginatos/química , Fosfatos de Cálcio/química , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Análise Espaço-Temporal , Ácido Glucurônico/química , Ácidos Hexurônicos/química , Concentração de Íons de Hidrogênio , Microscopia de Contraste de Fase , Imagem Óptica , Análise Espectral RamanRESUMO
UNLABELLED: Due to their large water content and structural similarities to the extracellular matrix, hydrogels are an attractive class of material in the tissue engineering field. Polymers capable of ionotropic gelation are of special interest due to their ability to form gels at mild conditions. In this study we have developed an experimental toolbox to measure the gelling kinetics of alginate upon crosslinking with calcium ions. A reaction-diffusion model for gelation has been used to describe the diffusion of calcium within the hydrogel and was shown to match experimental observations well. In particular, a single set of parameters was able to predict gelation kinetics over a wide range of gelling ion concentrations. The developed model was used to predict the gelling time for a number of geometries, including microspheres typically used for cell encapsulation. We also demonstrate that this toolbox can be used to spatiotemporally investigate the formation and evolution of mineral within the hydrogel network via correlative Raman microspectroscopy, confocal laser scanning microscopy and electron microscopy. STATEMENT OF SIGNIFICANCE: Hydrogels show great promise in cell-based tissue engineering, however new fabrication and modification methods are needed to realize the full potential of hydrogel based materials. The inclusion of an inorganic phase is one such approach and is known to affect both cell-material interactions and mechanical properties. This article describes the development of a correlative experimental approach where gel formation and mineralization has been investigated with spatial and temporal resolution by applying Raman microspectroscopy, optical and electron microscopy and a reaction-diffusion modeling scheme. Modeling allows us to predict gelling kinetics for other geometries and sizes than those investigated experimentally. Our experimental system enables non-destructive study of composite hydrogel systems relevant for, but not limited to, applications within bone tissue engineering.
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Alginatos/química , Hidrogéis/química , Análise Espaço-Temporal , Ácido Glucurônico/química , Ácidos Hexurônicos/química , Concentração de Íons de Hidrogênio , Cinética , Microscopia , Minerais/química , Modelos Teóricos , Análise Espectral Raman , Fatores de TempoRESUMO
Due to high solubility and fast resorption behaviour under physiological conditions, brushite (CaHPO4â 2H2O, calcium monohydrogen phosphate dihydrate, dicalcium phosphate dihydrate) has great potential in bone regeneration applications, both in combination with scaffolds or as a component of calcium phosphate cements. The use of brushite in combination with hydrogels opens up possibilities for new cell-based tissue engineering applications of this promising material. However, published preparation methods of brushite composites, in which the mineral phase is precipitated within the hydrogel network, fail to offer the necessary degree of control over the mineral phase, content and distribution within the hydrogel matrix. The main focus of this study is to address these shortcomings by determining the precise fabrication parameters needed to prepare composites with controlled composition and properties. Composite alginate microbeads were prepared using a counter-diffusion technique, which allows for the simultaneous crosslinking of the hydrogel and precipitation of an inorganic mineral phase. Reliable nucleation of a desired mineral phase within the alginate network proved more challenging than simple aqueous precipitation. This was largely due to ion transport within the hydrogel producing concentration gradients that modified levels of supersaturation and favoured the nucleation of other phases such as hydroxyapatite and octacalcium phosphate, which would otherwise not form. To overcome this, the incorporation of brushite seed crystals resulted in good control during the mineral phase, and by adjusting the number of seeds and amount of precursor concentration, the amount of mineral could be tuned. The material was characterised with a range of physical techniques, including scanning electron microscopy, powder x-ray diffraction and Rietveld refinement, Fourier transform infrared spectroscopy, and thermogravimetric analysis, in order to assess the mineral morphology, phase and amount within the organic matrix. The mineral content of the composite material converted from brushite into hydroxyapatite when submerged in simulated body fluid, indicating possible bioactivity. Additionally, initial cell culture studies revealed that both the material and the synthesis procedure are compatible with cells relevant to bone tissue engineering.
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Alginatos/química , Substitutos Ósseos/síntese química , Fosfatos de Cálcio/química , Hidrogéis/química , Osteoblastos/citologia , Osteoblastos/fisiologia , Células 3T3 , Animais , Líquidos Corporais/química , Proliferação de Células/fisiologia , Sobrevivência Celular/fisiologia , Cristalização/métodos , Ácido Glucurônico/química , Ácidos Hexurônicos/química , Teste de Materiais , CamundongosRESUMO
Pancreas disease (PD), caused by salmonid alphavirus subtype 3 (SAV3), emerged in Norwegian aquaculture in the 1980s and is now endemic along the south-western coast. In 2011, the first cases of PD caused by marine salmonid alphavirus subtype 2 (SAV2) were reported. This subtype has spread rapidly among the fish farms outside the PD-endemic zone and is responsible for disease outbreaks at an increasing numbers of sites. To describe the geographical distribution of salmonid alphavirus (SAV), and to assess the time and site of introduction of marine SAV2 to Norway, an extensive genetic characterization including more than 200 SAV-positive samples from 157 Norwegian marine production sites collected from May 2007 to December 2012 was executed. The first samples positive for marine SAV2 originated from Romsdal, in June 2010. Sequence analysis of the E2 gene revealed that all marine SAV2 included in this study were nearly identical, suggesting a single introduction into Norwegian aquaculture. Further, this study provides evidence of a separate geographical distribution of two subtypes in Norway. SAV3 is present in south-western Norway, and marine SAV2 circulates in north-western and Mid-Norway, a geographical area which since 2010 constitutes the endemic zone for marine SAV2.
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Infecções por Alphavirus/veterinária , Alphavirus/genética , Doenças dos Peixes/virologia , Proteínas Virais/genética , Proteínas Virais/metabolismo , Alphavirus/classificação , Infecções por Alphavirus/transmissão , Infecções por Alphavirus/virologia , Animais , Aquicultura , Doenças dos Peixes/transmissão , Tipagem Molecular , Noruega , Filogenia , SalmonidaeRESUMO
Pancreas disease (PD) caused by salmonid alphavirus (SAV) has a significant negative economic impact in the salmonid fish farming industry in northern Europe. Until recently, only SAV subtype 3 was present in Norwegian fish farms. However, in 2011, a marine SAV 2 subtype was detected in a fish farm outside the PD-endemic zone. This subtype has spread rapidly among fish farms in mid-Norway. The PD mortality in several farms has been lower than expected, although high mortality has also been reported. In this situation, the industry and the authorities needed scientific-based information about the virulence of the marine SAV 2 strain in Norway to decide how to handle this new situation. Atlantic salmon post-smolts were experimentally infected with SAV 2 and SAV 3 strains from six different PD cases in Norway. SAV 3-infected fish showed higher mortality than SAV 2-infected fish. Among the SAV 3 isolates, two isolates gave higher mortality than the third one. At the end of the experiment, fish in all SAV-infected groups had significantly lower weight than the uninfected control fish. This is the first published paper on PD to document that waterborne infection produced significantly higher mortality than intraperitoneal injection.