Fiber Scaffolds of Poly (glycerol-dodecanedioate) and its Derivative via Electrospinning for Neural Tissue Engineering

Peripheral nerves have demonstrated the ability to bridge gaps of up to 6 mm. Peripheral Nerve System injury sites beyond this range need autograft or allograft surgery. Central Nerve System cells do not allow spontaneous regeneration due to the intrinsic environmental inhibition. Although stem cell therapy seems to be a promising approach towards nerve repair, it is essential to use the distinct three-dimensional architecture of a cell scaffold with proper biomolecule embedding in order to ensure that the local environment can be controlled well enough for growth and survival. Many approaches have been developed for the fabrication of 3D scaffolds, and more recently, fiber-based scaffolds produced via the electrospinning have been garnering increasing interest, as it offers the opportunity for control over fiber composition, as well as fiber mesh porosity using a relatively simple experimental setup. All these attributes make electrospun fibers a new class of promising scaffolds for neural tissue engineering. Therefore, the purpose of this doctoral study is to investigate the use of the novel material PGD and its derivative PGDF for obtaining fiber scaffolds using the electrospinning. The performance of these scaffolds, combined with neural lineage cells derived from ESCs, was evaluated by the dissolvability test, Raman spectroscopy, cell viability assay, real time PCR, Immunocytochemistry, extracellular electrophysiology, etc. The newly designed collector makes it possible to easily obtain fibers with adequate length and integrity. The utilization of a solvent like ethanol and water for electrospinning of fibrous scaffolds provides a potentially less toxic and more biocompatible fabrication method. Cell viability testing demonstrated that the addition of gelatin leads to significant improvement of cell proliferation on the scaffolds. Both real time PCR and Immunocytochemistry analysis indicated that motor neuron differentiation was achieved through the high motor neuron gene expression using the metabolites approach. The addition of Fumaric acid into fiber scaffolds further promoted the differentiation. Based on the results, this newly fabricated electrospun fiber scaffold, combined with neural lineage cells, provides a potential alternate strategy for nerve injury repair.

Fiber scaffolds of poly (glycerol-dodecanedioate) and its derivative via electrospinning for neural tissue engineering

Peripheral nerves have demonstrated the ability to bridge gaps of up to 6 mm. Peripheral Nerve System injury sites beyond this range need autograft or allograft surgery. Central Nerve System cells do not allow spontaneous regeneration due to the intrinsic environmental inhibition. Although stem cell therapy seems to be a promising approach towards nerve repair, it is essential to use the distinct three-dimensional architecture of a cell scaffold with proper biomolecule embedding in order to ensure that the local environment can be controlled well enough for growth and survival. Many approaches have been developed for the fabrication of 3D scaffolds, and more recently, fiber-based scaffolds produced via the electrospinning have been garnering increasing interest, as it offers the opportunity for control over fiber composition, as well as fiber mesh porosity using a relatively simple experimental setup. All these attributes make electrospun fibers a new class of promising scaffolds for neural tissue engineering. Therefore, the purpose of this doctoral study is to investigate the use of the novel material PGD and its derivative PGDF for obtaining fiber scaffolds using the electrospinning. The performance of these scaffolds, combined with neural lineage cells derived from ESCs, was evaluated by the dissolvability test, Raman spectroscopy, cell viability assay, real time PCR, Immunocytochemistry, extracellular electrophysiology, etc. The newly designed collector makes it possible to easily obtain fibers with adequate length and integrity. The utilization of a solvent like ethanol and water for electrospinning of fibrous scaffolds provides a potentially less toxic and more biocompatible fabrication method. Cell viability testing demonstrated that the addition of gelatin leads to significant improvement of cell proliferation on the scaffolds. Both real time PCR and Immunocytochemistry analysis indicated that motor neuron differentiation was achieved through the high motor neuron gene expression using the metabolites approach. The addition of Fumaric acid into fiber scaffolds further promoted the differentiation. Based on the results, this newly fabricated electrospun fiber scaffold, combined with neural lineage cells, provides a potential alternate strategy for nerve injury repair.^

Substratos para uso em telhados verdes: Avaliação da retenção hídrica e qualidade da água de escoamento.

Telhados verdes são uma alternativa interessante para mitigar o risco de enchentes dada a enorme área de telhados não utilizada das superfícies impermeáveis nas áreas urbanas. Graças a sua capacidade de armazenagem de água, os telhados verdes podem reduzir significativamente o pico de escoamento dos eventos de maior pluviosidade. Investigações sobre a composição de substratos baseados em materiais locais e projetos adequados para regiões climáticas tropicais são menos frequentes. Vegetação e substrato são elementos de um telhado verde que precisam ser adaptados para cada microclima e não universalizados. O objetivo deste estudo foi avaliar o desempenho de diferentes composições de substratos baseados em solo local, biomassa de coco, condicionador de solo e componentes comerciais, com a finalidade de maximizar a capacidade de retenção da água de chuva e diminuir necessidades de manutenção. Um pré-ensaio de colunas avaliou a capacidade de retenção hídrica e a relação peso seco x peso úmido de 15 composições de substrato. As composições com melhores resultados constituíram os substratos S1(15% solo + 55% coco + 30%componentes comerciais), S2 (30% solo + 40% coco + 30%componentes comerciais) e S3 (60% solo + 10% coco + 30%componentes comerciais). A caracterização físico-química dos substratos, solo e fibra de coco foi realizada. Em seguida um teste de colunas avaliou a capacidade de retenção hídrica dos substratos sob duas condições de precipitação: uma leve (8,77 mm/h); e outra mais forte (42,0 mm/h). Os resultados apontaram que os substratos S2 e S3 apresentaram melhores resultados de retenção para ambas as intensidades de precipitação. Observou-se que S1, que apresentou melhor capacidade de retenção no pré-ensaio, teve desempenho inferior aos demais o que pode ser atribuído à maior concentração de fibra de coco na sua composição e o consequente surgimento de caminhos preferenciais ao longo do perfil da coluna, por onde a água escoou mais rapidamente. Em eventos de precipitação mais leve, os substratos reteram de 60 a 100% do total aplicado. Quando se aplicou uma intensidade de precipitação mais forte, a faixa de retenção ficou entre 40% e 59%. No entanto, as variáveis analisadas para avaliar a qualidade da água de escoamento dos substratos (pH, CE, P, NO3, NH4, Ca, Mg, Cu, Fe, Mn, Zn, Cr, Co, Ni) ficaram acima dos valores comumente encontrados na literatura, indicando que as composições aqui adotadas para os substratos podem implicar em uma fonte de poluição hídrica. Os substratos para uso em telhados verdes apresentados neste estudo atenderam seu objetivo quanto à retenção hídrica, mas a qualidade da água percolada torna seu uso inviável até o momento. Verificou-se a necessidade de estudar mais profundamente a qualidade da água lixiviada por cada componente dos substratos, individualmente, a fim de identificar as fontes dos elementos que presentes em concentrações elevadas tornam-se poluentes. Pode ser considerada a remoção de algum (s) dos componentes presentes na composição para se atingir um nível satisfatório de qualidade da água de escoamento.

Effects of forage source and extruded linseed supplementation on methane emissions from growing dairy cattle of differing body weights

Changes in diet carbohydrate amount and type (i.e., starch vs. fiber) and dietary oil supplements can affect ruminant methane emissions. Our objectives were to measure methane emissions, whole-tract digestibility, and energy and nitrogen utilization from growing dairy cattle at 2 body weight (BW) ranges, fed diets containing either high maize silage (MS) or high grass silage (GS), without or with supplemental oil from extruded linseed (ELS). Four Holstein-Friesian heifers aged 13 mo (BW range from start to finish of 382 to 526 kg) were used in experiment 1, whereas 4 lighter heifers aged 12 mo (BW range from start to finish of 292 to 419 kg) were used in experiment 2. Diets were fed as total mixed rations with forage dry matter (DM) containing high MS or high GS and concentrates in proportions (forage:concentrate, DM basis) of either 75:25 (experiment 1) or 60:40 (experiment 2), respectively. Diets were supplemented without or with ELS (Lintec[AU1: Add manufacturer name and location.]; 260 g of oil/ kg of DM) at 6% of ration DM. Each experiment was a 4 × 4 Latin square design with 33-d periods, with measurements during d 29 to 33 while animals were housed in respiration chambers. Heifers fed MS at a heavier BW (experiment 1) emitted 20% less methane per unit of DM intake (yield) compared with GS (21.4 vs. 26.6, respectively). However, when repeated with heifers of a lower BW (experiment 2), methane yield did not differ between the 2 diets (26.6 g/kg of DM intake). Differences in heifer BW had no overall effect on methane emissions, except when expressed as grams per kilogram of digestible organic matter (OMD) intake (32.4 vs. 36.6, heavy vs. light heifers). Heavier heifers fed MS in experiment 1 had a greater DM intake (9.4 kg/d) and lower OMD (755 g/kg), but no difference in N utilization (31% of N intake) compared with heifers fed GS (7.9 kg/d and 799 g/kg, respectively). Tissue energy retention was nearly double for heifers fed MS compared with GS in experiment 1 (15 vs. 8% of energy intake, respectively). Heifers fed MS in experiment 2 had similar DM intake (7.2 kg/d) and retention of energy (5% of intake energy) and N (28% of N intake), compared with GS-fed heifers, but OMD was lower (741 vs. 765 g/kg, respectively). No effect of ELS was noted on any of the variables measured, irrespective of animal BW, and this was likely due to the relatively low amount of supplemental oil provided. Differences in heifer BW did not markedly influence dietary effects on methane emissions. Differences in methane yield were attributable to differences in dietary starch and fiber composition associated with forage type and source.

Slow component of VO2 kinetics: the effect of training status, fibre type, UCP3 mRNA and citrate synthase activity

Examines the relationship between the magnitude of the relative slow component (SC) of pulmonary oxygen uptake VO[sub 2], citrate synthase activity, UCP2 and UCP3 mRNA levels and muscle fiber composition in both endurance-trained and recreationally active subjects. Magnitude of the relative SC of the Tr group; Indicators of aerobic fitness; High negative correlations between the magnitude of the relative SC and citrate synthase activity and VO[sub 2] peak.

Impaired muscle Ca2+ and K+ regulation contribute to poor exercise performance post-lung transplantation

Lung transplant recipients (LTx) exhibit marked peripheral limitations to exercise. We investigated whether skeletal muscle Ca2+ and K+ regulation might be abnormal in eight LTx and eight healthy controls. Peak oxygen consumption and arterialized venous plasma [K+] (where brackets denote concentration) were measured during incremental exercise. Vastus lateralis muscle was biopsied at rest and analyzed for sarcoplasmic reticulum Ca2+ release, Ca2+ uptake, and Ca2+-ATPase activity rates; fiber composition; Na+-K+-ATPase (K+-stimulated 3-O-methylfluorescein phosphatase) activity and content ([3H]ouabain binding sites); as well as for [H+] and H+-buffering capacity. Peak oxygen consumption was 47% less in LTx (P < 0.05). LTx had lower Ca2+ release (34%), Ca2+ uptake (31%), and Ca2+-ATPase activity (25%) than controls (P < 0.05), despite their higher type II fiber proportion (LTx, 75.0 ± 5.8%; controls, 43.5 ± 2.1%). Muscle [H+] was elevated in LTx (P < 0.01), but buffering capacity was similar to controls. Muscle 3-O-methylfluorescein phosphatase activity was 31% higher in LTx (P < 0.05), but [3H]ouabain binding content did not differ significantly. However, during exercise, the rise in plasma [K+]-to-work ratio was 2.6-fold greater in LTx (P < 0.05), indicating impaired K+ regulation. Thus grossly subnormal muscle calcium regulation, with impaired potassium regulation, may contribute to poor muscular performance in LTx.

Effect of streptozotocin-induced diabetes on glycogen resynthesis in fasted rats post-high-intensity exercise

It has recently been shown that food intake is not essential for the resynthesis of the stores of muscle glycogen in fasted animals recovering from high-intensity exercise. Because the effect of diabetes on this process has never been examined before, we undertook to explore this issue. To this end, groups of rats were treated with streptozotocin (60 mg/kg body mass ip) to induce mild diabetes. After 11 days, each animal was fasted for 24 h before swimming with a lead weight equivalent to 9% body mass attached to the tail. After exercise, the rate and the extent of glycogen repletion in muscles were not affected by diabetes, irrespective of muscle fiber composition. Consistent with these findings, the effect of exercise on the phosphorylation state of glycogen synthase in muscles was only minimally affected by diabetes. In contrast to its effects on nondiabetic animals, exercise in fasted diabetic rats was accompanied by a marked fall in hepatic glycogen levels, which, surprisingly, increased to preexercise levels during recovery despite the absence of food intake.

MicroRNA-23a has minimal effect on endurance exercise-induced adaptation of mouse skeletal muscle.

Skeletal muscles contain several subtypes of myofibers that differ in contractile and metabolic properties. Transcriptional control of fiber-type specification and adaptation has been intensively investigated over the past several decades. Recently, microRNA (miRNA)-mediated posttranscriptional gene regulation has attracted increasing attention. MiR-23a targets key molecules regulating contractile and metabolic properties of skeletal muscle, such as myosin heavy-chains and peroxisome proliferator-activated receptor gamma, coactivator 1 alpha (PGC-1α). In the present study, we analyzed the skeletal muscle phenotype of miR-23a transgenic (miR-23a Tg) mice to explore whether forced expression of miR-23a affects markers of mitochondrial content, muscle fiber composition, and muscle adaptations induced by 4 weeks of voluntary wheel running. When compared with wild-type mice, protein markers of mitochondrial content, including PGC-1α, and cytochrome c oxidase complex IV (COX IV), were significantly decreased in the slow soleus muscle, but not the fast plantaris muscle of miR-23a Tg mice. There was a decrease in type IId/x fibers only in the soleus muscle of the Tg mice. Following 4 weeks of voluntary wheel running, there was no difference in the endurance exercise capacity as well as in several muscle adaptive responses including an increase in muscle mass, capillary density, or the protein content of myosin heavy-chain IIa, PGC-1α, COX IV, and cytochrome c. These results show that miR-23a targets PGC-1α and regulates basal metabolic properties of slow but not fast twitch muscles. Elevated levels of miR-23a did not impact on whole body endurance capacity or exercise-induced muscle adaptations in the fast plantaris muscle.

Efeito da Temperatura Ambiente e da Restrição Alimentar sobre o Desempenho e a Composição de Fibras Musculares Esqueléticas de Frangos de Corte

O objetivo desta pesquisa foi estudar os efeitos da temperatura ambiente e da restrição alimentar sobre o desempenho e a composição do músculo flexor longo do hálux de frangos de corte. Trezentos e vinte e quatro pintos machos da linhagem Ross, com cinco dias de idade, foram distribuídos em um delineamento em parcelas subdivididas, considerando os tratamentos principais no esquema fatorial 3x3 inteiramente casualizado (três programas de alimentação: ad libitum, restrição precoce - 8 a 14 dias e restrição tardia - 29 a 35 dias; três níveis de temperatura: termoneutra, calor e frio). Os tratamentos secundários foram as idades das aves (seis idades: 7, 14, 21, 28, 35 e 42 dias). Não houve interação entre programa de alimentação e temperatura ou entre programa de alimentação, temperatura e idade para as características de desempenho dos frangos. Independentemente do programa de alimentação, houve efeito de temperatura para peso, ganho de peso, consumo de ração e conversão alimentar. O programa de alimentação afetou o peso e o ganho de peso das aves. Houve interação entre programa de alimentação e idade e entre temperatura e idade para peso vivo ao abate e peso do músculo da perna direita. Estas interações não foram significativas para peso do músculo da perna esquerda, área da secção transversal do músculo e relação peso do músculo/peso vivo ao abate. Conclui-se que a temperatura ambiente afeta o desempenho dos frangos de corte, porém não altera o número, diâmetro e freqüência de fibras musculares no músculo flexor longo do hálux. A restrição alimentar precoce pode ser adotada como prática de manejo, sem que se observem alterações do desempenho na idade de abate e nem na composição das fibras musculares esqueléticas dos frangos de corte.

Morfologia das fibras musculares esqueléticas de frangos de corte criados nos sistemas confinado e semi-confinado

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação das características físico-químicas e microbiológicas dos componentes fibrosos obtidos da moagem úmida de grãos de milho

Fibers are non-digestible materials by human organism, and they are insoluble in specific conditions. The corn fiber has been used as an alternative to enrich the food products for human consumption, in order to add nutritious value to this co-product. This study aimed at determining the chemical and fractions components of corn fiber, extracted by semi-wet milling in laboratory (LabF), and by semi-humid milling in industrial process (IndF); and microbiological analyses were perform in the last fraction. The fiber composition differed significantly among these two products as to moisture, ash, lipids, proteins, insoluble dietary fiber, neutral detergent fiber (NDF), acid detergent fiber (ADF), cellulose and lignin contents. The sample IndF showed microbiological parameters in compliance with the specifications for consumption. The investigated corn fiber may effectively contribute to increase the fiber intake in the diet. As the industrially processed fiber (IndF) showed compliant microbiological parameters with the specifications for food, this component might be incorporated into the food products formulation.

Locomotor performance of closely related Tropidurus species: relationships with physiological parameters and ecological divergence

Tropidurid lizards have colonized a variety of Brazilian open environments without remarkable morphological variation, despite ecological and structural differences among habitats used. This study focuses on two Tropidurus sister-species that, despite systematic proximity and similar morphology, exhibit great ecological divergence and a third ecologically generalist congeneric species providing an outgroup comparison. We quantified jumping capacity and sprint speed of each species on sand and rock to test whether ecological divergence was also accompanied by differences in locomotor performance. Relevant physiological traits possibly associated with locomotor performance metabolic scopes and fiber type composition, power output and activity of the enzymes citrate synthase, pyruvate kinase and lactate dehydrogenase of the iliofibularis muscle - were also compared among the three Tropidurus species. We found that the two sister-species exhibited remarkable differences in jumping performance, while Tropidurus oreadicus, the more distantly related species, exhibited intermediate values. Tropidurus psamonastes, a species endemic to sand dunes, exhibited high absolute sprint speeds on sand, jumped rarely and possessed a high proportion of glycolytic fibers and low activity of citrate synthase. The sister-species Tropidurus itambere, endemic to rocky outcrops, performed a large number of jumps and achieved lower absolute sprint speed than T. psamonastes. This study provides evidence of rapid divergence of locomotor parameters between sister-species that use different substrates, which is only partially explained by variation in physiological parameters of the iliofibularis muscle.

MicroRNA-23a has minimal effect on endurance exercise-induced adaptation of mouse skeletal muscle

Skeletal muscles contain several subtypes of myofibers that differ in contractile and metabolic properties. Transcriptional control of fiber-type specification and adaptation has been intensively investigated over the past several decades. Recently, microRNA (miRNA)-mediated posttranscriptional gene regulation has attracted increasing attention. MiR-23a targets key molecules regulating contractile and metabolic properties of skeletal muscle, such as myosin heavy-chains and peroxisome proliferator-activated receptor gamma, coactivator 1 alpha (PGC-1α). In the present study, we analyzed the skeletal muscle phenotype of miR-23a transgenic (miR-23a Tg) mice to explore whether forced expression of miR-23a affects markers of mitochondrial content, muscle fiber composition, and muscle adaptations induced by 4 weeks of voluntary wheel running. When compared with wild-type mice, protein markers of mitochondrial content, including PGC-1α, and cytochrome c oxidase complex IV (COX IV), were significantly decreased in the slow soleus muscle, but not the fast plantaris muscle of miR-23a Tg mice. There was a decrease in type IId/x fibers only in the soleus muscle of the Tg mice. Following 4 weeks of voluntary wheel running, there was no difference in the endurance exercise capacity as well as in several muscle adaptive responses including an increase in muscle mass, capillary density, or the protein content of myosin heavy-chain IIa, PGC-1α, COX IV, and cytochrome c. These results show that miR-23a targets PGC-1α and regulates basal metabolic properties of slow but not fast twitch muscles. Elevated levels of miR-23a did not impact on whole body endurance capacity or exercise-induced muscle adaptations in the fast plantaris muscle.

Optimization study of the structural HACFRA composition (Self Compacting Concrete Steel Fiber Reinforced)

The interest of HACFRA (self compacting concrete reinforced with steel fibers), is the combination of the residual strength increase and cracking decrease compared to plain concrete by the introduction of steel fibers in the mass with the advantages of the self-compacting. The paper presents an analysis of the influence of different components of the HACRFA and provides their selection, refered to the granular skeleton and to different steel fiber types and amount, in order to obtain an optimization of its features and structural behavior.