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Organoids are miniature, organ-like structures derived from stem cells. While techniques for developing organoids for different tissues have advanced in the past decade, it has remained a challenge to simultaneously grow two different tissues into a single functional organoid. We have recently developed a 3D neuromuscular organoid that allows the simultaneous generation of spinal neurons and skeletal muscles. Here we discuss their future applications in modeling neuromuscular disorders.

RESUMO Concentrados com alta proteína bruta e minerais e com menor concentração de energia poderiam ser utilizados em menores quantidades no balanceamento de dietas com volumosos tropicais. Assim, objetivou-se avaliar o consumo de matéria seca, a resposta produtiva e econômica de vacas da raça Girolando mantidas em pastagem de Tifton 85 durante o período de seca, as quais receberam diferentes concentrações de suplementação proteica. Foram utilizadas 12 vacas da raça Girolando em lactação, com produção de leite média inicial de 18,00±1,615kg/dia. Os animais foram distribuídos em três quadrados latinos (4x4), constituídos de quatro tratamentos e quatro períodos experimentais, com duração de 15 dias cada. Os tratamentos consistiram de uma dieta controle, apenas sal mineral e três níveis de suplementação proteica com 40; 50 e 60% de proteína bruta (PB) sendo fornecidas nas quantidades de 2,5kg/animal/dia. Observou-se que os fornecimentos de suplementos proteicos proporcionaram um efeito substitutivo da forragem em relação à suplementação mineral. O uso de suplementação concentrada com teor de 40% de proteína bruta foi mais eficiente em termos produtivos, econômicos e de produção de proteína bruta no leite. ABSTRACT Concentrates with high crude protein and minerals and lower energy concentration could be used in smaller quantities in balancing diets with tropical bulk. The objective of this study was to evaluate the dry matter intake, the productive and economical response of Girolando cows kept in Tifton 85 pasture during dry season receiving different concentrations of protein supplementation. Twelve lactating Girolando cows with initial mean milk yield of 18.00±1.615kg/day were used. The animals were distributed in three Latin squares (4x4), consisting of four treatments and four experimental periods, each lasting 15 days. The treatments were composed of control diet with only mineral salt, and three levels of protein supplementation with 40, 50, and 60% crude protein (CP) being supplied in the amounts of 2.5kg/animal/day. It was found that the protein supplement supplies provided a substitutive effect of forage relative to mineral supplementation. The use of concentrated supplementation with 40% crude protein content was more efficient in terms of production, production of crude protein in milk and economically.

The biotechnological and industrial uses of thermostable and organic solvent-tolerant enzymes are extensive and the investigation of such enzymes from microbiota present in oil reservoirs is a promising approach. Searching sequence databases for esterases from such microbiota, we have identified in silico a potentially secreted esterase from Acetomicrobium hydrogeniformans, named AhEst. The recombinant enzyme was produced in E. coli to be used in biochemical and biophysical characterization studies. AhEst presented hydrolytic activity on short-acyl-chain p-nitrophenyl ester substrates. AhEst activity was high and stable in temperatures up to 75 °C. Interestingly, high salt concentration induced a significant increase of catalytic activity. AhEst still retained ~ 50% of its activity in 30% concentration of several organic solvents. Synchrotron radiation circular dichroism and fluorescence spectroscopies confirmed that AhEst displays high structural stability in extreme conditions of temperature, salinity, and organic solvents. The enzyme is a good emulsifier agent and is able to partially reverse the wettability of an oil-wet carbonate substrate, making it of potential interest for use in enhanced oil recovery. All the traits observed in AhEst make it an interesting candidate for many industrial applications, such as those in which a significant hydrolytic activity at high temperatures is required.

El proyecto multidisciplinar TREASURE tiene como objetivo principal caracterizar la diversidad de razas locales y sistemas de producción para productos sostenibles y de alta calidad. Uno de los principales objetivos del proyecto consiste en la caracterización de la diversidad genética de una serie de razas locales, incluidas la alentejana y la ibérica, mediante el uso de variantes genéticas/ polimorfismos que son variaciones en la secuencia del ADN entre los individuos de una población. Para llevar a cabo esta caracterización, se genotiparon variantes genéticas en individuos ibéricos, alentejanos y en otras 18 razas autóctonas de porcino. Teniendo en cuenta los resultados obtenidos en el presente estudio, los cuales muestran una cercanía genética muy grande en las variantes genéticas analizadas, podría deducirse que las razas ibéricas y alentejana fueran estirpes pertenecientes a una misma raza, más que razas independientes por sí misma. Dentro del proyecto TREASURE se están realizando análisis utilizando un mayor número de información genética a lo largo de todo el genoma para corroborar los resultados mostrados en el presente estudio. Estos resultados junto con la información para el resto de poblaciones será presentada en un futuro.

In order to assess the productive performance of the Ribatejano (RI) pig, resulting from a cross between Alentejano (AL) and Bísaro (BI) breeds, castrated male pigs AL, BI, ALxBI and BIxAL were studied within the framework of the TREASURE project. Ten pigs from each genotype, raised in traditional free-range system and fed commercial diets ad libitum, were slaughtered at ~65 kg live weight (LW). Data obtained show that BI, ALxBI and BIxAL attained slaughter weight faster (P

Enzymes isolated from thermophilic organisms found in oil reservoirs can find applications in many fields, including the oleochemical, pharmaceutical, bioenergy, and food/dairy industries. In this study, in silico identification and recombinant production of an esterase from the extremophile bacteria Petrotoga mobilis (designated PmEst) were performed. Then biochemical, bioinformatics and structural characterizations were undertaken using a combination of synchrotron radiation circular dichroism (SRCD) and fluorescence spectroscopies to correlate PmEst stability and hydrolytic activity on different substrates. The enzyme presented a high Michaelis-Menten constant (KM 0.16 mM) and optimum activity at ~55°C for p-nitrophenyl butyrate. The secondary structure of PmEst was preserved at acid pH, but not under alkaline conditions. PmEst was unfolded at high concentrations of urea or guanidine through apparently different mechanisms. The esterase activity of PmEst was preserved in the presence of ethanol or propanol and its melting temperature increased ~8°C in the presence of these organic solvents. PmEst is a mesophilic esterase with substrate preference towards short-to medium-length acyl chains. The SRCD data of PmEst is in agreement with the prediction of an α/β protein, which leads us to assume that it displays a typical fold of esterases from this family. The increased enzyme stability in organic solvents may enable novel applications for its use in synthetic biology. Taken together, our results demonstrate features of the PmEst enzyme that indicate it may be suitable for applications in industrial processes, particularly, when the use of polar organic solvents is required.

Para este trabalho, foram utilizados lombos (matéria-prima) obtidos em 3 sistemas de produção diferentes, com matrizes produtivas bem caracterizadas e conhecidas: 1) porcos de raça Alentejana engordados em montanheira (PAMA), 2)porcos de raça alentejana Alentejana (DOP) e 3) porcos industriais (PI). Os lombos frescos foram identificados e sujeitos ao mesmo processo tecnológico de elaboração, na fábrica Companhia Alentejana de Enchidos Tradicionais. Após o processo de cura, a composição química centesimal dos lombos frescos foi determinada, bem como o seu teor de colagénio total e o teor de cloretos (sal). Os lombos de porco Alentejano (PAM e DOP) registaram uma marcada diferença de composição química relativamente aos lombos de PI, principalmente no teor de gordura intramuscular (lípidos neutros). O maior teor de gordura intramuscular dos lombos de porco Alentejano afetou o teor de humidade e de proteína, mas na avaliação sensorial da suculência, os lombos de porco Alentejano mereceram melhor avaliação, confirmando a importância da gordura intramuscular nas características de textura dos lombo curado. Estes dados sugerem mesmo que a sua influência é superior à da concentração em colagénio total, que apesar de superior nos lombos de porco Alentejano, não afetou negativamente a sensação de suculência

Este trabalho teve como objetivo a utilização de óleo de girassol rico em ácido oleico na alimentação de suínos Alentejanos destinados à produção de carne para consumo em fresco. O ensaio foi realizado em condições de exploração. Utilizaram-se 12 suínos castrados alojados num parque com uma área de 3 Ha. Com um peso vivo médio de 60 Kg os animais foram divididos em dois grupos de 6 suínos. O grupo 1 (controle) foi alimentado com uma dieta baseada em cereais (3 150 Kcal ED, 14% PB e 1,7% EE). O grupo 2 (alto oleico) foi alimentado com uma dieta com 6% de óleo de girassol rico em ácido oleico (3375 Kcal ED, 14%PB e 7,9%EE). Os suínos foram alimentados individualmente, tendo-se utilizado um nível alimentar de cerca de 85% do AD Libitum. Procedeu-se, diariamente, ao controle individual do alimento fornecido e ingerido e, semanalmente, à pesagem dos animais. A ração distribuída diariamente foi calculada em função do seu valor energético e ajustada ao peso médio do grupo, para que a ingestão energética fosse igual. Os animais foram abatidos com um peso médio de 100 Kg e determinadas características da carcaça. O ganho médio diário foi 556 e 551 g/dia, respetivamente no grupo controle e alto oleico. Os suínos alimentados com a dieta normal ingeriram mais 6,6% de alimento originando um índice de conversão alimentar de 4,37, contra 4,21, no grupo alto oleico, embora a diferença não tenha sido estatisticamente significativas. Entre o grupo controle e o grupo alto oleico não se observaram diferenças significativas no peso de abate (99,75 vs 99,00 kg), no peso da carcaça quente (78,87 vs 78,42 kg), no rendimento de carcaça (78,87 vs 78,42%), na espessura média de gordura subcutânea dorsal (4,95 vs 4,85 cm), na espessura média do músculo longíssimos dorsi (3,09 vs 3,23 cm) e no rendimento em peças nobres (43,27 vs 43,77%). Os resultados deste ensaio demonstram que a utilização de óleo de girassol alto oleico na dieta de porcos alentejanos destinados à produção de carne para consumo em fresco não afeta negativamente as performances de crescimento, nem as características da carcaça.

The aim of this work was to determine the relationship between physical measures from the subcutaneous tissue and Longissimus dorsi (LD) muscle (area, depth, and width measured between the 3rd and 4th lumbar vertebrae, at the last rib, and between the 3rd and 4th last ribs) and the chemical composition of LD at 70, 80, 90, 100, and 110 kg LW. The content of water, protein, neutral and polar lipids, total and soluble collagen, and total pigments, were determined. Globally, the measures taken and the chemical composition were not affected between 70 and 110 kg, except for the LD depth and width at the 3rd-4th lumbar vertebrae. At 70 kg, the LD depth was greater than at 110 kg (3.77 vs. 2.75 cm, P

The present work studied the evolution of fatty acid composition in the dorsal subcutaneous fat of free-range Alentejano pigs fed on pasture and acorns. Twenty pigs with a body weight (BW) of ~90 kg were placed in a 2 ha pasture (Lolium westerwoldium and Trifolium incarnatum) and fed daily with 4 kg of acorns. Animals were submitted to biopsies near tail insertion with ~90, ~100, and at slaughter, ~110 kg BW. The fat samples obtained were analysed by GC/FID to determine the fatty acid composition. An increase of only 20 kg body mass based on pasture and acorn consumption led to: (i) carcasses with C18:1 content similar to the minimal value stipulated by market demands for Alentejano pigs bred in free-range conditions; and (ii) contents of C16:0 and C18:0 were within the limits for the carcass-quality assessment. Nevertheless, the contents in C18:2 remained slightly above the 9.8% limit.

This trial was undertaken to determine the effect of the rearing system (intensive vs. extensive) over the content of antioxidants (alpha-tocopherol) in the meat of Alentejano pigs. Sixteen Alentejano pigs with an average body weight (BW) of 60 kg were divided in two groups: Group I (n=10) was allocated in open-air individual pens, while Group II (n=6) was maintained in a 3 ha natural pasture. Both groups were fed a commercial diet (3100 kcal ED, 14% CP, and 20 mg/kg alpha-tocopherol) at 85% ad libitum. All pigs were killed at 100 kg BW and carcasses were refrigerated (24 h at 4ºC). Samples of semimembranosus muscle were obtained, stored under vacuum, and frozen (-30ºC) until analysis. The content in alpha-tocopherol was estimated by HPLC. Results show a significant effect (P

Neste trabalho experimental procedeu-se à medição da espessura e à caracterização histológica de lesões ateroscleróticas, em suínos de raça Alentejana, procurando-se estabelecer uma relação entre estas e os valores dos parâmetros sanguíneos associados ao desenvolvimento deste processo patológico. As concentrações plasmáticas de triacilgliceróis, fosfolípidos, colesterol total, colesterol livre, LDLc e HDLc foram determinadas por métodos enzimáticos. Foram também feitas análises histopatológicas a amostras da artéria coronária esquerda. Os animais foram divididos em 2 grupos de 6 indivíduos cada: Grupo I, com elevada colesterolémia (4,25 mmol/L) e Grupo II, com níveis normais (2,53 mmol/L). Os valores do ganho médio diário (GMD) dos dois grupos foram semelhantes. Os animais do Grupo I apresentaram valores significativamente mais elevados (P=0,001) para: colesterol total, colesterol livre, colesterol esterificado e LDLc. A área de lesão foi significativamente superior (P=0,05) no Grupo I. Verificou-se uma relação linear entre a área de lesão (fases iniciais do tipo I e II) e os teores plasmáticos de colesterol total, de LDLc e de colesterol livre, o que sugere a influência destes parâmetros na dimensão da área de lesão. Os resultados deste trabalho sugerem que os suínos de raça Alentejana podem desenvolver lesões ateroscleróticas ao longo do seu ciclo de vida, tal como o observado em humanos e outras raças de suínos. Estas lesões estão associadas a hipercolesterolemia que poderão ser devidas a mutações genéticas em apolipoproteínas, sistemas enzimáticos ou receptores. São necessários estudos futuros, quer a nível histológico quer a nível de biologia molecular, para um maior aprofundamento do conhecimento das lesões ateroscleróticas em suínos de raça Alentejana., Revista de Ciências Agrárias, vol. 30 n.º 1 (2007)

A dieta mediterrânica é rica em alimentos funcionais. As leguminosas, um dos alimentos-chave desta dieta, têm visto o seu papel na prevenção de dislipidémias, diabetes e cancro do cólon mencionado por muitos autores. O efeito do consumo de leguminosas na redução da colesterolémia deve-se a diferentes nutrientes e fitoquímicos, tais como: i) proteína; ii) lípidos, particularmente a componente polinsaturada e monoinsaturada; iii) fibra, especialmente a fracção solúvel; iv) saponinas; e v) fitosteróis. O consumo da soja tem sido relacionado com tais efeitos benéficos, mas dados recentes obtidos na Universidade de Évora demonstraram que leguminosas como a ervilha e o tremoço de folhas estreitas também apresentam elevado potencial funcional na regulação do colesterol sanguíneo. O seu consumo levou a reduções de 30% na colesterolémia de animais experimentais, via redução do colesterol das LDL. A utilização destas leguminosas como alimentos funcionais e/ou como fornecedoras de fitoquímicos com potencial preventivo e terapêutico é promissora e poderá constituir uma maisvalia e uma fonte extra de rendimento para os agricultores que se dediquem ao cultivo destas espécies., Revista de Ciências Agrárias, vol. 30 n.º 1 (2007)

This work has evaluated the influence of body weight (BW) on some plasma parameters of Alentejano pigs. Thirty castrated Alentejano pigs (~35 kg BW) were allocated in individual pens and fed a commercial diet (0.85x ad libitum). The pigs were slaughtered in groups (n=5) with an average BW of 42.2, 70.9, 80.2, 90.1, 100.5, and 110.2 kg. Plasma samples obtained at slaughter were analysed. Glucose and triacylglycerol levels decreased (P=0.05) with increasing BW, suggesting that during growth Alentejano pigs increased the use of glucose and triacylglycerols as energy and substrate for fat synthesis and deposition. Total cholesterol increased (P=0.05) with BW, due to an increase (P=0.05) in the low‑ density lipoprotein (LDL) cholesterol level, following the trends reported in ageing mammals. The Alentejano pig, with a high lipogenic activity, could be a valuable model for studies of the cholesterol metabolism.

This work aimed to evaluate the effects of body weight and rearing system on total lipids and cholesterol concentration of fresh m. semimembranosus and subcutaneous fat from Alentejano pigs.

Membro da Comissão Organizadora. Este curso decorreu na Universidade de Évora e foi creditado pela FELASA (Federation of European Laboratory Animal Science Associations) (nº 020/08) seguindo as suas recomendações para o treino de indivíduos habilitados a trabalhar com animais de laboratório, categoria C.

To study the effects of slaughter weight on growth and carcass characteristics of Alentejano Pig thirty pigs were used. With 20 kg Lw animals were allocated in individual pens at outdoor and fed a commercial diet (15% CP, 3100 Kcal DE) offered at 85% Ad Libitum. Pigs were sequentially slaughtered in groups (n=5) with an average BW of 40, 70, 80, 90, 100 and 110 kg. The growing period, feed intake, feed: gain and feed cost per Kg gain increased significantly (P

Partial and dynamically reconfigurable SRAM-based FPGAs (Field Programmable Gate Arrays) enable the implementation of reconfigurable systems hosting several applications simultaneously, which share the available resources according to the functional requirements that are present at any given moment. Time and space sharing strategies enabled the concept of virtual hardware, supporting the concurrent implementation of applications which would otherwise require far more complex resources. However, the performance of these applications (e.g. execution speed and reliability, activation delay) is directly influenced by the efficiency of the management strategies that allocate the logic space to the various functions that are waiting to be activated (each function requiring a specific amount of logic resources). Because the activation requests are in most cases not predictable, all resource allocation decisions have to be made online. The consequences of such working contexts are twofold: 􀂃 All FPGA resources must be tested regularly, to exclude malfunctioning due to the allocation of faulty elements. Since the process of launching / halting active functions takes place asynchronously at any given moment, an online concurrent test scheme is the only way of ensuring reliable system operation and predictable fault detection latency; 􀂃 As the resources are allocated to functions and later released, many small “islands” of free resources are created. If these areas become too small, they will be left unused due to routing restrictions. The defragmentation of the FPGA logic space must therefore be carried out regularly, to avoid the wasting of logic resources. This paper presents a non-intrusive solution for the concurrent replication of active logic blocks (i.e. logic blocks that are being used to implement part of an active function), transferring their functionality to fault-free resources that are available in the FPGA logic space. This replication scheme is then used as the core of an online concurrent test strategy that scans the complete FPGA, reusing the available 1149.1 test infrastructure to carry out a structural test of each logic block that has just been released. The overhead of the proposed solution, in terms of the number of configurable logic resources required for its implementation, as well as its performance (e.g. the resulting fault detection latency), are quantified. Further to the test aspects, an online concurrent defragmentation strategy based on the same replication scheme is also proposed. A rearrangement of the available logic space is carried out by selectively releasing active logic blocks, with the objective of enforcing the adjacency of those blocks that share the implementation of a common function, and the creation of wider pools of logic resources that may be used to implement new functions.

A study was conducted to determine the effects of ileo-rectal anastomosis (IR) and protein source on apparent digestibility in growing pigs. Intact animals fed casein diet had significantly higher CP and NDF digestibilities than IR pigs. Intact animals fed soybean diet presented significantly higher DM, OM, CP, NDF, ADF and GE digestibilities than IR pigs. Intact pigs fed casein diet presented significantly higher DM, OM, CP, NDF and GE, and lower TL digestibilities than those fed soybean diet. IR pigs fed casein diet had higher DM, OM, CP, NDF, ADF and GE digestibility values than the ones fed soybean diet.

A field plot study was conducted over three years, which compared six exotic grass species planted in July 1994, in terms of fry matter (DM) yield, crude protein and in vitro digestibility. The genotypes evaluated were Paspalum dilatatum, Paspalum urvillei, Pennisetum clandestinum, Pennisetum purpureum, Setaria sphacelata var. sericea x S. sphacelata var. splendida (Splenda) and Setaria splendida. Three of them, namely P. dilatatum, P. clandestinum, and Splenda established well ans rapidly after planting. Only by the third year was a good stand obtained with Setaria plendida and this genotype supplied only one harvest, in November 1996. Over all three years, total yield tended to be highest with Splenda, which produced 3126, 8438 and 14022 kh ha-1 of total DM yield for 1994, 1995 and 1996 respectively. Of the species which produced harvestable yield in every year, P. clandestinum had DM yields, but seemed to have the most consistent yields for the harvests in a year. This species showed good characteristics for permanent pastures due to its prostate growth habit and ease of spread over the soil. Forage nutritive value, as measured by crude protein concentration and in vitro dry matter digestibility, was greatest in the species which was one of the most difficult to establish: P. purpureum.

This study was done to evaluate the ensilability of the tropical grass Splenda (Setaria sphacelata var. sericea x S. sphacelata var. splendida) and the effect of several treatments on the characteristics and quality of the silage obtained. Irrigated grass was chopped ( 4 cm) and submitted to four treatments prior to ensiling: control treatment (chopped green grass) (T1); 24 h pre-wilted grass (T2); green grass with 85% formic acid (4 l ADD-F®/t); and green grass with Ecosyl® (7.5 g/t). Results showed that T4 was the best treatment. T1, T2 and T3 presented worse results, with higher values of pH, ammonia nitrogen, NDF and ADF and lower values of lactic acid and in vitro dry matter and organic matter digestibility.