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Changes in elemental and mineralogical composition, and lead speciation, of air pollution control residue (APCR) from municipal solid waste incineration, due to treatment by water washing, were investigated in this work and are reported in the context of a review of the literature. Water washing was shown to substantially modify the nature of APCR by: 1) removing 23% dry mass soluble salts to disagglomerate particles and significantly reduce concentrations of the associated major elements, and increase concentrations of insoluble matrix elements and potential pollutants; and 2) respeciating elements to form new phases. X-ray absorption near edge spectroscopy (XANES) showed that the 500 mg/kg of Pb in raw and washed APCR were comprised mainly of Pb-glass, with some PbSO4, and small amounts of PbO and PbCl2. Semi-quantitative linear combination fitting suggests that the glass in the APCR may be unstable and release Pb under the alkaline pH of water washing, to reprecipitate as PbO. Chemical analysis suggests that some Pb may be removed by washing. Scientific understanding of the composition of raw and washed APCR, and particularly the speciation of potentially toxic metals, such as Zn and Pb, can help in developing effective element recovery and residue treatment, utilization or disposal strategies.
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Despite the high innate regenerative capacity of bone, large osseous defects fail to heal and remain a clinical challenge. Healing such defects requires the formation of large amounts of bone in an environment often rendered hostile to osteogenesis by damage to the surrounding soft tissues and vasculature. In recent years, there have been intensive research efforts directed towards tissue engineering and regenerative approaches designed to overcome this multifaceted challenge. In this paper, we describe and critically evaluate the state-of-the-art approaches to address the various components of this intricate problem. The discussion includes (i) the properties of synthetic and natural scaffolds, their use in conjunction with cell and growth factor delivery, (ii) their vascularisation, (iii) the potential of gene therapies and (iv) the role of the mechanical environment. In particular, we present a critical analysis of where the field stands, and how it can move forward in a coordinated fashion.
Assuntos
Regeneração Óssea/fisiologia , Osso e Ossos/patologia , Engenharia Tecidual/métodos , Animais , Sistemas de Liberação de Medicamentos , Terapia Genética , Humanos , Alicerces Teciduais/químicaRESUMO
The process of new blood vessel formation is critical in tissue development, remodeling and regeneration. Modular tissue engineering approaches have been developed to enable the bottom-up assembly of more complex tissues, including vascular networks. In this study, collagen-fibrin composite microbeads (100-300 µm in diameter) were fabricated using a water-in-oil emulsion technique. Human endothelial cells and human fibroblasts were embedded directly in the microbead matrix at the time of fabrication. Microbead populations were characterized and cultured for 14 days either as free-floating populations or embedded in a surrounding fibrin gel. The collagen-fibrin matrix efficiently entrapped cells and supported their viability and spreading. By 7 days in culture, endothelial cell networks were evident within microbeads, and these structures became more prominent by day 14. Fibroblasts co-localized with endothelial cells, suggesting a pericyte-like function, and laminin deposition indicated maturation of the vessel networks over time. Microbeads embedded in a fibrin gel immediately after fabrication showed the emergence of cells and the coalescence of vessel structures in the surrounding matrix by day 7. By day 14, inosculation of neighboring cords and prominent vessel structures were observed. Microbeads pre-cultured for 7 days prior to embedding in fibrin gave rise to vessel networks that emanated radially from the microbead by day 7, and developed into connected networks by day 14. Lumen formation in endothelial cell networks was confirmed using confocal sectioning. These data show that collagen-fibrin composite microbeads support vascular network formation. Microbeads embedded directly after fabrication emulated the process of vasculogenesis, while the branching and joining of vessels from pre-cultured microbeads resembled angiogenesis. This modular microtissue system has utility in studying the processes involved in new vessel formation, and may be developed into a therapy for the treatment of ischemic conditions.
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This position paper summarises a vision of how cell-based therapies can be applied clinically to regenerate bone, as well as the steps needed to narrow the gap between that vision and clinical reality. It is a result of the presentations and discussion of the "Cell Therapy for Bone Repair" breakout session at the AO Foundation Symposium "Where Science Meets Clinics" in Davos, Switzerland from September 5-7, 2013. Participants included leaders from science, medicine, and industry from around the world. The session included clinical and scientific presentations, as well as an extended discussion among participants. Bone tissue has an innate regenerative capacity that in most cases allows functional healing at damage sites. However, there are a number of serious conditions in which bone does not fully heal and the result is significant morbidity. The clinical need for new therapies is clear, and the breakout session participants were enthusiastic about the potential impact on cell-based therapies for bone repair in the clinic. However, they also recognised the significant challenges that face the development of commercially viable cell therapy products. This paper outlines a vision in which patient selection is based on expected therapeutic outcome to create a consistently successful, cost-effective, cell-based therapy for bone repair. The need for a more complete understanding of bone repair, a better infrastructure for preclinical studies, and the need for collaboration among stakeholders is discussed.
Assuntos
Regeneração Óssea , Transplante de Células-Tronco/métodos , Pesquisa Translacional Biomédica , Animais , HumanosRESUMO
Collagen type I is a widely used natural biomaterial that has found utility in a variety of biological and medical applications. Its well-characterized structure and role as an extracellular matrix protein make it a highly relevant material for controlling cell function and mimicking tissue properties. Collagen type I is abundant in a number of tissues, and can be isolated as a purified protein. This review focuses on hydrogel biomaterials made by reconstituting collagen type I from a solubilized form, with an emphasis on in vitro studies in which collagen structure can be controlled. The hierarchical structure of collagen from the nanoscale to the macroscale is described, with an emphasis on how structure is related to function across scales. Methods of reconstituting collagen into hydrogel materials are presented, including molding of macroscopic constructs, creation of microscale modules and electrospinning of nanoscale fibers. The modification of collagen biomaterials to achieve the desired structures and functions is also addressed, with particular emphasis on mechanical control of collagen structure, creation of collagen composite materials and crosslinking of collagenous matrices. Biomaterials scientists have made remarkable progress in rationally designing collagen-based biomaterials and in applying them both to the study of biology and for therapeutic benefit. This broad review illustrates recent examples of techniques used to control collagen structure and thereby to direct its biological and mechanical functions.
Assuntos
Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Colágeno/química , Colágeno/farmacologia , Animais , Colágeno/ultraestrutura , Reagentes de Ligações Cruzadas/farmacologia , Humanos , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Hidrogel de Polietilenoglicol-Dimetacrilato/farmacologiaRESUMO
Cardiac fibroblasts are exposed to both cyclic strain and interstitial fluid flow in the myocardium. The balance of these stimuli is affected by fibrotic scarring, during which the fibroblasts transition to a myofibroblast phenotype. The present study investigates the mechanisms by which cardiac fibroblasts seeded in three-dimensional (3D) collagen gels differentiate between strain and fluid flow. Neonatal cardiac fibroblast-seeded 3D collagen gels were exposed to interstitial flow and/or cyclic strain and message levels of collagens type I and III, transforming growth factor ß1 (TGF-ß1), and α-smooth muscle actin (α-SMA) were assessed. Flow was found to significantly increase and strain to decrease expression of myofibroblast markers. Corresponding immunofluorescence indicated that flow and strain differentially regulated α-SMA protein expression. The effect of flow was inhibited by exposure to losartan, an angiotensin II type 1 receptor (AT1R) blocker, and by introduction of shRNA constructs limiting AT1R expression. Blocking of TGF-ß also inhibited the myofibroblast transition, suggesting that flow-mediated cell signaling involved both AT1R and TGF-ß1. Reduced smad2 phosphorylation in response to cyclic strain suggested that TGF-ß is part of the mechanism by which cardiac fibroblasts differentiate between strain-induced and flow-induced mechanical stress. Our experiments show that fluid flow and mechanical deformation have distinct effects on cardiac fibroblast phenotype. Our data suggest a mechanism in which fluid flow directly acts on AT1R and causes increased TGF-ß1 expression, whereas cyclic strain reduces activation of smad proteins. These results have relevance to the pathogenesis and treatment of heart failure.
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Líquido Extracelular/metabolismo , Miofibroblastos/metabolismo , Receptor Tipo 1 de Angiotensina/metabolismo , Estresse Mecânico , Fator de Crescimento Transformador beta1/metabolismo , Bloqueadores do Receptor Tipo 1 de Angiotensina II/farmacologia , Animais , Células Cultivadas , Losartan/farmacologia , Miofibroblastos/citologia , Miofibroblastos/efeitos dos fármacos , Ratos , Ratos Sprague-Dawley , Receptor Tipo 1 de Angiotensina/biossínteseRESUMO
Portland cement (CEM I) and ground granulated blast furnace slag (ggbs) have been used to treat air pollution control (APC) residues from an energy-from-waste plant burning municipal solid waste. Stabilised/solidified (s/s) products were prepared with binder additions ranging from 10 to 50 wt.% of total dry mass and water/solids ratios between 0.40 and 0.80. Monolithic leach tests (EA NEN 7375:2004) indicated that 50% binder additions were necessary to meet the UK monolithic Waste Acceptance Criteria (monWAC) for Pb and Zn, and previous work indicated that chloride leaching exceeded WAC even at this binder addition. Lower binder additions (20 and 10%) did not sufficiently reduce leaching of Pb. Although the monWAC are based on an assumption that leaching is diffusion-controlled, evaluation of leaching mechanisms indicates that more complex processes than diffusion occur for s/s APC residues.
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Poluição do Ar , Metais/química , Difração de Raios XRESUMO
Multispectral three-dimensional (3D) imaging provides spatial information for biological structures that cannot be measured by traditional methods. This work presents a method of tracking 3D biological structures to quantify changes over time using graph theory. Cell-graphs were generated based on the pairwise distances, in 3D-Euclidean space, between nuclei during collagen I gel compaction. From these graphs quantitative features are extracted that measure both the global topography and the frequently occurring local structures of the "tissue constructs." The feature trends can be controlled by manipulating compaction through cell density and are significant when compared to random graphs. This work presents a novel methodology to track a simple 3D biological event and quantitatively analyze the underlying structural change. Further application of this method will allow for the study of complex biological problems that require the quantification of temporal-spatial information in 3D and establish a new paradigm in understanding structure-function relationships.
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Técnicas de Cultura de Células/métodos , Imageamento Tridimensional/métodos , Células-Tronco Mesenquimais/citologia , Modelos Biológicos , Contagem de Células , Colágeno Tipo I/farmacologia , Fluorescência , Humanos , Hidrogel de Polietilenoglicol-Dimetacrilato/farmacologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Microscopia Confocal , Fatores de TempoRESUMO
In the treatment of pulmonary diseases the inhalation of aerosols plays a key role - it is the preferred route of drug delivery in asthma, chronic obstructive pulmonary disease (COPD) and cystic fibrosis. But, in contrast to oral and intravenous administration drug delivery to the lungs is controlled by additional parameters. Beside its pharmacology the active agent is furthermore determined by its aerosol characteristics as particle diameter, particle density, hygroscopicity and electrical charge. The patient related factors like age and stage of pulmonary disease will be additionally affected by the individual breathing pattern and morphometry of the lower airways. A number of these parameters with essential impact on the pulmonary drug deposition can be influenced by the performance of the inhalation system. Therefore, the optimization of nebulisation technology was a major part of aerosol science in the last decade. At this time the control of inspiration volume and air flow as well as the administration of a defined aerosol bolus was in the main focus. Up to date a more efficient and a more targeted pulmonary drug deposition - e.g., in the alveoli - will be provided by novel devices which also allow shorter treatment times and a better reproducibility of the administered lung doses. By such means of precise dosing and drug targeting the efficacy of inhalation therapy can be upgraded, e.g., the continuous inhalation of budesonide in asthma. From a patients' perspective an optimized inhalation manoeuvre means less side effects, e.g., in cystic fibrosis therapy the reduced oropharyngeal tobramycin exposure causes fewer bronchial irritations. Respecting to shorter treatment times also, this result in an improved quality of life and compliance. For clinical trials the scaling down of dose variability in combination with enhanced pulmonary deposition reduces the number of patients to be included and the requirement of pharmaceutical compounds. This review summarises principles and advances of individualised controlled inhalation (ICI) as offered by the AKITA inhalation system.
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Ensaios Clínicos como Assunto , Nebulizadores e Vaporizadores , Assistência ao Paciente , Administração por Inalação , Aerossóis , Humanos , Pulmão/metabolismoRESUMO
The extracellular matrix provides structural and organizational cues for tissue development and defines and maintains cellular phenotype during cell fate determination. Multipotent mesenchymal stem cells use this matrix to tightly regulate the balance between their differentiation potential and self-renewal in the native niche. When understood, the mechanisms that govern cell-matrix crosstalk during differentiation will allow for efficient engineering of natural and synthetic matrices to specifically direct and maintain stem cell phenotype. This work identifies the discoidin domain receptor 1 (DDR1), a collagen activated receptor tyrosine kinase, as a potential link through which stem cells sense and respond to the 3D organization of their extracellular matrix microenvironment. DDR1 is dependent upon both the structure and proteolytic state of its collagen ligand and is specifically expressed and localized in three dimensional type I collagen culture. Inhibition of DDR1 expression results in decreased osteogenic potential, increased cell spreading, stress fiber formation and ERK1/2 phosphorylation. Additionally, loss of DDR1 activity alters the cell-mediated organization of the naïve type I collagen matrix. Taken together, these results demonstrate a role for DDR1 in the stem cell response to and interaction with three dimensional type I collagen. Dynamic changes in cell shape in 3D culture and the tuning of the local ECM microstructure, directs crosstalk between DDR1 and two dimensional mechanisms of osteogenesis that can alter their traditional roles.
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Stabilisation/solidification with cementitious or pozzolanic binders (S/S) is an option for reducing leachability of contaminants from residual, predominantly inorganic, industrial wastes and contaminated soils before disposal or reuse. Treatment by S/S is complicated by the fact that the presence of impurities, such as the contaminants and bulk matrix components present in industrial wastes, can have deleterious effects on cements. Therefore, careful laboratory development and testing of S/S formulations are required prior to full-scale application, to avoid technology failures, including problems with handling and contaminant retention. An understanding of cement chemistry and contaminant immobilisation mechanisms has been used to propose a series of test methods and performance thresholds for use in efficient evaluation of the treatability of industrial wastes by S/S, and optimising S/S formulations: measurement of stabilised/solidified product workability, bleeding and setting time (for flowable mixtures) or Proctor compaction (for compactable mixtures), together with unconfined compressive strength, leachability in a batch extraction with distilled water, and hydraulic conductivity.
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Cimentação/métodos , Resíduos Industriais , Compostos Inorgânicos/análise , Compostos Inorgânicos/química , Eliminação de Resíduos/métodos , Concentração de Íons de Hidrogênio , Metais Pesados/química , SolubilidadeRESUMO
Portland cement (CEMI) was used to solidify air pollution control (APC) residues from an energy-from-waste plant burning municipal solid waste. APC residue/CEMI mixes were prepared with CEMI additions ranging from 0 to 50 weight% (wt%) of total dry mass and water/solids ratios between 0.40 and 0.80. Isothermal conduction calorimetry was used to assess the effect of APC residues on the hydration of CEMI. Although up to 30wt% additions of APC residues accelerated CEMI hydration, the total heat of hydration during the initial 98h was significantly reduced. Higher levels of APC residues severely inhibited CEMI hydration. The consistence, setting time, compressive strength, porosity and chloride leaching characteristics of the solidified products were determined. As might be expected, increasing the CEMI addition and reducing the water content resulted in increased compressive strengths. All mixes achieved compressive strengths greater than 1MPa at 7 and 28days but only 50wt% samples did not show significant strength reduction when tested after immersion in water. Monolithic leaching tests indicated low physical immobilisation of chloride in the CEMI solidified APC residues, with chloride leaching in excess of relevant UK landfill waste acceptance criteria (WAC). The results of this study show that greater than 50% CEMI additions would be required to effectively treat APC residues to meet current WAC limits.
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Poluentes Atmosféricos/química , Cloretos/química , Conservação dos Recursos Naturais , Materiais de Construção , Resíduos Industriais , Teste de Materiais , Eliminação de Resíduos , Cidades , Força Compressiva , Microscopia Eletrônica de Varredura , Porosidade , Temperatura , Fatores de Tempo , Água/químicaRESUMO
Neural network analysis was used to construct models of unconfined compressive strength (UCS) as a function of mix composition using existing data from literature studies of Portland cement containing real industrial wastes. The models were able to represent the known non-linear dependency of UCS on curing time and water content, and generalised from the literature data to find relationships between UCS and quantities of five waste types. Substantial decreases in UCS were caused by all wastes; except for EAF dust, the effect was nonlinear with the greatest decrease caused initially by approx. 12% plating sludge, 40% foundry dust, 58% other ash, and 72% MSWI fly ash by mass of dry product. It appears that the maximum waste additions used in modelling may approximate the practical limits of waste additions used in modelling may approximate the practical limits of waste addition to Portland cement, i.e., 50% plating sludge or EAF dust, 64% foundry dust, 92% other ash, and 85% MSWI fly ash by mass of dry product. The laboratory was found to be a key predictive variable and acted as a surrogate for laboratory-specific variables related to cement composition, strength and hardening class, product mixing and preparation details, laboratory conditions, and testing details. While the neural network modelling approach has been shown to be feasible, development of better models would require larger data sets with more complete information regarding laboratory-specific variables and waste composition.
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Conservação dos Recursos Naturais , Materiais de Construção , Resíduos Industriais , Eliminação de Resíduos , Força Compressiva , Previsões , Incineração , Teste de Materiais , Redes Neurais de ComputaçãoRESUMO
Neural network models, to predict the leachate pH for single batch extraction leaching tests conducted on Portland cement pastes containing pure compounds, were constructed using existing data from the literature. The models were able to represent the known non-linear dependency of pH on acid addition, and were used to show that Cu increases, and Zn and NO3- decrease, the leachate pH for addition of 8 meq acid/g dry cement (to achieve a mid-alkaline pH). Ba, Cd, Cr(III), Ni, Pb, Cl- and OH- had no detectable effect on the acid neutralisation capacity (ANC) of the cement pastes in the concentration ranges investigated. The laboratory where testing was conducted was found to be an important predictive variable, which acted as a surrogate variable for laboratory specific variables that were not adequately reported in the literature, such as cement characteristics, sample preparation details, and leaching test and pH measurement details. This work has shown that development of good empirical predictive models for solidified product leachate pH is feasible, and is limited only by the availability of data.
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Manufaturas , Metais Pesados/química , Redes Neurais de Computação , Eliminação de Resíduos/métodos , Poluição Ambiental , Previsões , Concentração de Íons de Hidrogênio , Teste de MateriaisRESUMO
The efficacy of clinical islet transplantation has been demonstrated with autografts, and although islet allografts have established insulin independence in a small number of IDDM patients, the treatment is confounded by the necessity of immunosuppression. the lack of donor tissue, and recurring islet immunogenicity. These limitations underscore a need to develop therapies to serve the large population of diabetic patients. Porcine islet xenotransplantation, together with a successful immune intervention strategy, may provide the necessary clinical alternative. However, a major obstacle in evaluating this approach has been the difficulty of obtaining adequate volumes of functional islet tissue from pigs. Donors of market weight are preferable to retired breeders due to their abundance, lower animal and husbandry costs. and are more suitable to meet regulatory guidelines for donor tissue for xenotransplantation. We describe a simple isolation procedure that following purification yields a mean of 350,000 IE, corresponding to 179 units of insulin and 1.8 mg of DNA with an islet purity and viability in excess of 85% (n = 317 isolations). In both short- and long-term cell cultures, porcine islets demonstrated glucose-responsive insulin secretion. However, this secretion is density dependent, which may have significant consequences in the development of immunoisolation technologies to support porcine islet xenotransplantation. Following implantation into diabetic nude mice, porcine islets remained functional in excess of 1 year. Implantation of a bioartificial pancreas containing porcine islets into pancreatectomized dogs provided significant clinical benefit with an improved diabetic condition. Finally, secretagogue-induced insulin release was demonstrated in vitro from these devices after removal from immunocompetent recipients. Immunohistochemical staining identified well-granulated islets following long-term implantation in both the rodent and canine models. This study demonstrates the ability to isolate porcine islets in clinically relevant numbers from market animals, which survive and remain functional for prolonged periods of time in an immune-deficient or immunoprotected environment.
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Transplante das Ilhotas Pancreáticas , Ilhotas Pancreáticas/citologia , Ilhotas Pancreáticas/fisiologia , Pâncreas Artificial , Animais , Peso Corporal , Separação Celular , Células Cultivadas , Diabetes Mellitus Tipo 1/cirurgia , Cães , Feminino , Glucose/farmacologia , Insulina/metabolismo , Secreção de Insulina , Ilhotas Pancreáticas/efeitos dos fármacos , Masculino , Estimulação Química , Suínos , Transplante HeterólogoRESUMO
As cell-based therapies receive approval for clinical evaluation and use, the development of reliable methods to quantify cell number and control the dose of therapy delivered is becoming increasingly important. An example is the determination of the number and volume of primary porcine hepatocytes used in an extracorporeal treatment for patients with liver disease. Conventional cell counting using optical microscopy was compared against two alternate methods to quantify isolated porcine hepatocytes: (1) automated cell counting using a commercially available particle characterization instrument, and (2) quantitation by cell mass. Methods were compared based on accuracy, precision, specificity, linear range, and ruggedness. The automated method delivered substantially improved accuracy, precision, and ruggedness when compared to the conventional optical method. It also provided valuable information about the size distribution of cell preparations, which often contained clumps of cells, and showed that processing steps such as cryopreservation can alter the size characteristics of a cell population. The automated method was also faster, and was well suited to use in a commercial manufacturing process. The mass-based method was simple and inexpensive, but suffered from nonlinearity at low cell concentrations. Automated cell quantitation using a commercially available particle characterization instrument proved to be the preferred method for obtaining accurate and consistent porcine hepatocyte counts in a timely manner.
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Contagem de Células/métodos , Transplante de Células , Transplante de Fígado , Fígado/citologia , Animais , SuínosRESUMO
BACKGROUND: Physiological blood pressure (BP) fluctuations with frequencies >0.1 Hz can override renal blood flow autoregulation. The influence of such immediate changes in renal perfusion pressure (RPP) on daily BP regulation, eg, via shear stress-stimulated liberation of renal endothelial NO, however, is unknown. Thus, we studied the effects of such RPP oscillations on renal function and on systemic BP during the onset of renal hypertension. METHODS AND RESULTS: Seven beagles (randomly assigned to each of the following protocols) were chronically instrumented for the measurement of systemic BP, RPP, and renal excretory function. An inflatable cuff was used to reduce and to oscillate RPP over 24 hours in the freely moving dog. Reducing RPP to 87+/-2 mm Hg diminished excretion of sodium and water and doubled plasma renin activity (PRA, n=7, P<0. 01) but had no significant effect on urinary nitrate excretion (n=6), a marker of NO generation. Superimposing 0.1-Hz oscillations (+/-10 mm Hg) onto the reduced RPP blunted hypertension, returned fluid excretion almost to control levels, and doubled renal sodium elimination. Nitrate excretion peaked at 8 hours, only to return to control values shortly thereafter. PRA, conversely, was significantly reduced during the last third of the experimental protocols. CONCLUSIONS: BP fluctuations transiently stimulate NO liberation and induce a reduction in PRA, which enhances 24-hour sodium and water excretion and markedly attenuates the acute development of renovascular hypertension.
Assuntos
Pressão Sanguínea/fisiologia , Hipertensão Renal/fisiopatologia , Rim/fisiologia , Animais , Cães , Feminino , Análise de Fourier , Rim/irrigação sanguínea , Óxido Nítrico/metabolismo , Circulação Renal , Renina/sangueRESUMO
Accurate and consistent measurement of tissue volume is critical to performing many types of islet research; however, conventional visual determination of isolated islet yields through a microscope is heavily operator dependent. An improved method of islet volume determination using digital image analysis (DIA) was developed to remove operator bias and automate the islet counting process. A series of 140 porcine islet isolations were used to evaluate the DIA method in three separate stages. In Stage 1 (n = 29 isolations), the conventional and DIA methods were correlated with two other independent islet quantitation methods: insulin extraction, and DNA extraction. It was found that volumes determined by DIA correlated more closely with insulin content and DNA content than did conventionally determined volumes. In Stages 2 and 3 (n = 54 and 57 isolations, respectively), it was shown that an increase in the number of fields analyzed by DIA did not significantly improve the quality of the correlations. Inclusion of very small tissue (<50 microm in diameter), which is ignored in the conventional protocol affected yields by less than 10% and did not significantly improve the correlation with insulin or DNA content. Quantitation of isolated islet tissue volume using DIA has been shown to be rapid, consistent, and objective. In the laboratory, use of this method as the standard for islet volume measurement will allow more meaningful comparison of experimental results between centers. In the clinic, its use will allow more accurate dosing of transplanted tissue.
Assuntos
Processamento de Imagem Assistida por Computador/métodos , Transplante das Ilhotas Pancreáticas , Ilhotas Pancreáticas/anatomia & histologia , Animais , DNA/análise , DNA/isolamento & purificação , Estudos de Avaliação como Assunto , Técnicas In Vitro , Insulina/análise , Insulina/isolamento & purificação , Ilhotas Pancreáticas/química , Reprodutibilidade dos Testes , SuínosRESUMO
UNLABELLED: Our aim was to elucidate why astronaut's exercise capacity after spaceflight is reduced. Therefore, the kinetics of oxygen uptake (VO2) as a measure for muscular aerobic capacity, as well as maximal oxygen uptake (VO2peak), and anaerobic threshold (PAT) as overall measures for exercise capacity were determined. Measurements of VO2peak and AT were restricted to pre- and postflight sessions. METHODS: Four crew-members of the D-2 mission (10 d) were cycling with steady state phases at 20 W and 80 W, followed by 450s of pseudo random binary sequence (PRBS) changes between 20 W and 80 W, and an incremental exercise test (10 W every 30 s) up to subjective exhaustion. Breath-by-breath VO2, VCO2, ventilation, HR, and blood pressure were continuously recorded. Blood lactate samples were drawn only during the incremental phase. The VO2 kinetics were determined by evaluation of the relationship between the workload and the instantaneous oxygen uptake of each subject. The cross-correlation function between both variables showed two characteristic items, the maximum as a measure for the muscular aerobic capacity, and its lag representing time consuming processes. RESULTS AND DISCUSSION: Each subject showed an individual characteristic of VO2 kinetics. In flight, no significant changes were detected compared to preflight data. Decreases in lags of cross-correlation function maxima and decreases in mean blood pressure during exercise indicate lowered blood volumes 2 d after the flight. Lowered blood volumes can explain the losses in exercise capacity. The maximum of the cross-correlation function did not change significantly which indicates unchanged muscular oxidative capacity.
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Limiar Anaeróbio/fisiologia , Tolerância ao Exercício/fisiologia , Voo Espacial , Adulto , Medicina Aeroespacial , Gasometria , Pressão Sanguínea , Teste de Esforço , Humanos , Ácido Láctico/sangue , Pessoa de Meia-Idade , Monitorização Fisiológica , Ventilação PulmonarRESUMO
We have developed a simple and efficient technique for automated parallel loading of >/=200 lanes on a 30 cm-wide gel in automated DNA sequencing, using porous filter materials and an associated manual or robotic system. The samples are loaded onto the teeth of a comb made of the porous material. The comb, with samples, is inserted directly above the straight edge of the polymerized gel. The samples are driven from the comb into the gel by the applied electrical field. A particularly advantageous aspect of this method is the elimination of the thin gel walls separating the sample wells in the standard gel loading technique. The time for sample loading is significantly reduced to a few minutes. The loading technique is applicable to horizontal or vertical systems, with standard or ultrathin gels.