RGS14 (414)-mediated prevention of an episodic memory loss: a study of molecular mechanism

A large proportion of human populations suffer memory impairments either caused by normal aging or afflicted by diverse neurological and neurodegenerative diseases. Memory enhancers and other drugs tested so far against memory loss have failed to produce therapeutic efficacy in clinical trials and thus, there is a need to find remedy for this mental disorder. In search for cure of memory loss, our laboratory discovered a robust memory enhancer called RGS14(414). A treatment in brain with its gene produces an enduring effect on memory that lasts for lifetime of rats. Therefore, current thesis work was designed to investigate whether RGS14(414) treatment can prevent memory loss and furthermore, explore through biological processes responsible for RGS-mediated memory enhancement. We found that RGS14(414) gene treatment prevented episodic memory loss in rodent models of normal aging and Alzheimer´s disease. A memory loss was observed in normal rats at 18 months of age; however, when they were treated with RGS14(414) gene at 3 months of age, they abrogated this deficit and their memory remained intact till the age of 22 months. In addition to normal aging rats, effect of memory enhancer treatment in mice model of Alzheimer´s disease (AD-mice) produced a similar effect. AD-mice subjected to treatment with RGS14(414) gene at the age of 2 months, a period when memory was intact, showed not only a prevention in memory loss observed at 4 months of age but also they were able to maintain normal memory after 6 months of the treatment. We posit that long-lasting effect on memory enhancement and prevention of memory loss mediated through RGS14(414) might be due to a permanent structural change caused by a surge in neuronal connections and enhanced neuronal remodeling, key processes for long-term memory formation. A neuronal arborization analysis of both pyramidal and non-pyramidal neurons in brain of RGS14(414)-treated rats exhibited robust rise in neurites outgrowth of both kind of cells, and an increment in number of branching from the apical dendrite of pyramidal neurons, reaching to almost three times of the control animals. To further understand of underlying mechanism by which RGS14(414) induces neuronal arborization, we investigated into neurotrophic factors. We observed that RGS14 treatment induces a selective increase in BDNF. Role of BDNF in neuronal arborization, as well as its implication in learning and memory processes is well described. In addition, our results showing a dynamic expression pattern of BDNF during ORM processing that overlapped with memory consolidation further support the idea of the implication of this neurotrophin in formation of long-term memory in RGS-animals. On the other hand, in studies of expression profiling of RGS-treated animals, we have demonstrated that 14-3-3ζ protein displays a coherent relationship to RGS-mediated ORM enhancement. Recent studies have demonstrated that the interaction of receptor for activated protein kinase 1 (RACK1) with 14-3-3ζ is essential for its nuclear translocation, where RACK1-14-3-3ζ complex binds at promotor IV region of BDNF and promotes an increase in BDNF gene transcription. These observations suggest that 14-3-3ζ might regulate the elevated level of BDNF seen in RGS14(414) gene treated animals. Therefore, it seems that RGS-mediated surge in 14-3-3ζ causes elevated BDNF synthesis needed for neuronal arborization and enhanced ORM. The prevention of memory loss might be mediated through a restoration in BDNF and 14-3-3ζ protein levels, which are significantly decreased in aging and Alzheimer’s disease. Additionally, our results demonstrate that RGS14(414) treatment could be a viable strategy against episodic memory loss

NIRS potential use for the determination of natural resources quality from dehesa (acorn and grass) in Montanera system for Iberian pigs.

NIRS technology has been used as an alternative to conventional methods to determinate the content of nutrients of acorns and grass from dehesa ecosystem. Dry matter (DM), crude fat (CF), crude protein (CP), starch, total phenolic compounds (TP), α-tocopherol, γ-tocopherol, fatty acids, neutral detergent fiber (NDF), total antioxidant activity (TAA) and total energy (TE) were determined by conventional methods for later development of NIRS predictive equations. The NIR spectrum of each sample was collected and for all studied parameters, a predictive model was obtained and external validated. Good prediction equations were obtained for moisture, crude fat, crude protein, total energy and γ-tocopherol in acorns samples, with high coefficients of correlation (1-VR) and low standard error of prediction (SEP) (1-VR=0.81, SEP=2.62; 1-VR=0.92, SEP=0.54; 1-VR=0.86, SEP=0.47; 1-VR=0.84, SEP=0.2; 1-VR=0.88, SEP=5.4, respectively) and crude protein, NDF, α-tocopherol and linolenic acid content in grass samples (1-VR=0.9, SEP=1.99; 1-VR=0.87, SEP=4.13; 1-VR=0.76, SEP=10.9; 1-VR=0.82, SEP=0.6, respectively). Therefore, these prediction models could be used to determinate the nutritional composition of Montanera natural resources

Computational analysis of a plant receptor interaction network

Trabajo fin de máster en Bioinformática y Biología ComputacionalIn all organisms, complex protein-protein interactions (PPI) networks control major biological functions yet studying their structural features presents a major analytical challenge. In plants, leucine-rich-repeat receptor kinases (LRR-RKs) are key in sensing and transmitting non-self as well as self-signals from the cell surface. As such, LRR-RKs have both developmental and immune functions that allow plants to make the most of their environments. In the model organism in plant molecular biology, Arabidopsis thaliana, most LRR-RKs are still represented by biochemically and genetically uncharacterized receptors. To fix this an LRR-based Cell Surface Interaction (CSI LRR ) network was obtained in 2018, a protein-protein interaction network of the extracellular domain of 170 LRR-RKs that contains 567 bidirectional interactions. Several network analyses have been performed with CSI LRR . However, these analyses have so far not considered the spatial and temporal expression of its proteins. Neither has it been characterized in detail the role of the extracellular domain (ECD) size in the network structure. Because of that, the objective of the present work is to continue with more in depth analyses with the CSI LRR network. This would provide important insights that will facilitate LRR-RKs function characterization. The first aim of this work is to test out the fit of the CSI LRR network to a scale-free topology. To accomplish that, the degree distribution of the CSI LRR network was compared with the degree distribution of the known network models of scale-free and random. Additionally, three network attack algorithms were implemented and applied to these two network models and the CSI LRR network to compare their behavior. However, since the CSI LRR interaction data comes from an in vitro screening, there is no direct evidence whether its protein-protein interactions occur inside the plant cells. To gain insight on how the network composition changes depending on the transcriptional regulation, the interaction data of the CSI LRR was integrated with 4 different RNA-Seq datasets related with the network biological functions. To automatize this task a Python script was written. Furthermore, it was evaluated the role of the LRR-RKs in the network structure depending on the size of their extracellular domain (large or small). For that, centrality parameters were measured, and size-targeted attacks performed. Finally, gene regulatory information was integrated into the CSI LRR to classify the different network proteins according to the function of the transcription factors that regulate its expression. The results were that CSI LRR fits a power law degree distribution and approximates a scale- free topology. Moreover, CSI LRR displays high resistance to random attacks and reduced resistance to hub/bottleneck-directed attacks, similarly to scale-free network model. Also, the integration of CSI LRR interaction data and RNA-Seq data suggests that the transcriptional regulation of the network is more relevant for developmental programs than for defense responses. Another result was that the LRR-RKs with a small ECD size have a major role in the maintenance of the CSI LRR integrity. Lastly, it was hypothesized that the integration of CSI LRR interaction data with predicted gene regulatory networks could shed light upon the functioning of growth-immunity signaling crosstalk

Surface topography of hydroxyapatite affects ROS17/2.8 cells response

Hydroxyapatite (HA) has been used in orthopedic, dental, and maxillofacial surgery as a bone substitute. The aim of this investigation was to study the effect of surface topography produced by the presence of microporosity on cell response, evaluating: cell attachment, cell morphology, cell proliferation, total protein content, and alkaline phosphatase (ALP) activity. HA discs with different percentages of microporosity (< 5%, 15%, and 30%) were confected by means of the combination of uniaxial powder pressing and different sintering conditions. ROS17/2.8 cells were cultured on HA discs. For the evaluation of attachment, cells were cultured for two hours. Cell morphology was evaluated after seven days. After seven and fourteen days, cell proliferation, total protein content, and ALP activity were measured. Data were compared by means of ANOVA and Duncan’s multiple range test, when appropriate. Cell attachment (p = 0.11) and total protein content (p = 0.31) were not affected by surface topography. Proliferation after 7 and 14 days (p = 0.0007 and p = 0.003, respectively), and ALP activity (p = 0.0007) were both significantly decreased by the most irregular surface (HA30). These results suggest that initial cell events were not affected by surface topography, while surfaces with more regular topography, as those present in HA with 15% or less of microporosity, favored intermediary and final events such as cell proliferation and ALP activity

O sistema dois-híbridos de Saccharomyces cerevisiae.

Variações e potencial para a análise funcional e montagem de redes de interações proteína-proteína

Utilización de la proteína dietaría por alevinos de la gamitana, Colossoma macropomum, alimentados con dietas isocalóricas

Un experimento fue conducido para evaluar los efectos de cinco niveles de proteína (25,27, 29, 31 y 33%) sobre el comportamiento productivo de alevinos de la gamitana, Colossoma macropomum Cuvier 1818, alimentados con dietas isocalóricas (2,7 kcal de ED/g). Los parámetros medidos fueron ganancia de peso (GP), conversión alimenticia (CA), proteína retenida (PR), razón de eficiencia proteica (REP) y energía retenida (ER). En la preparación de las dietas experimentales se utilizaron como fuentes de proteína harina de anchoveta y harina de torta de soya y como fuentes de energía maíz amarillo duro, subproducto de trigo y aceite de pescado. Se encontraron diferencias significativas (P&lt;0,05) entre tratamientos para los parámetros GP, PR, REP y ER. No se encontraron diferencias significativas para la CA. Los mejores rendimientos fueron obtenidos cuando las gamitanas fueron alimentadas con niveles dietarios de 25, 27 y 33% de proteína cruda. También se encontró que a medida que se elevó el nivel de proteína de la dieta, la REP decreció significativamente. Tomando en cuenta el costo de la proteína en la dieta, se concluye que la alimentación de la gamitana con niveles dietarios de 25 ó 27% de proteína cruda y 2,7 kcal de ED/g, garantizarán su exitoso crecimiento

In vivo evaluation of the protein quality of lyophilized egg white as substitute for casein in elaboration of an experimental diet based on AIN93.

El objetivo del estudio fue evaluar in vivo la calidad de la proteína de la clara de huevo liofilizada como sustituta de la caseína en la dieta AIN93. Se separaron ratones Wistar machos&nbsp;según las dietas: caseína (CS), clara de huevo (EW) y sin proteína (PF). Se evaluó el crecimiento somático y la ingesta de alimentos. Se midieron la relación de eficiencia de proteína, la relación neto de proteína, el perfil bioquímico, los pesos de los órganos y de los tejidos. La relación de eficacia de proteína muestra que EW presenta un alto valor biológico. Sin embargo, EW mostró un crecimiento menor que el CS. El peso relativo de los órganos y la creatinina se elevaron en el grupo EW. Por lo tanto, EW promueve la reducción del crecimiento, a pesar de la mayor ingesta.El objetivo del estudio fue evaluar in vivo la calidad de la proteína de la clara de huevo liofilizada como sustituta de la caseína en la dieta AIN93. Se separaron ratones Wistar machos&nbsp;según las dietas: caseína (CS), clara de huevo (EW) y sin proteína (PF). Se evaluó el crecimiento somático y la ingesta de alimentos. Se midieron la relación de eficiencia de proteína, la relación neto de proteína, el perfil bioquímico, los pesos de los órganos y de los tejidos. La relación de eficacia de proteína muestra que EW presenta un alto valor biológico. Sin embargo, EW mostró un crecimiento menor que el CS. El peso relativo de los órganos y la creatinina se elevaron en el grupo EW. Por lo tanto, EW promueve la reducción del crecimiento, a pesar de la mayor ingest

Avaliação in vivo da qualidade protéica do champignon do Brasil (Agaricus brasiliensis Wasser et al.).

Objetivo: O trabalho aqui descrito trata da avaliação de uma dieta experimental contendo Champingnon do Brasil (Agaricus brasiliensis) como fonte de proteína em um modelo experimental de ratos. Métodos: Para este propósito, foram selecionados 24 ratos Wistar machos, recém desmamados (21 dias) divididos em 3 grupos de 8 animais cada, que foram alimentados com uma dieta padrão de caseína, ou com uma dieta experimental de proteína de Agaricus brasiliensis ambas contendo 10% de proteína e isoenergéticas ou ainda, com uma dieta com muito baixo teor de proteína. O ensaio biológico foi realizado em 28 dias, ao longo dos quais se determinou a concentração de nitrogênio na urina e nas fezes, além dos cálculos do Quociente de Eficiência Alimentar (ganho de peso dividido pelo consumo de dieta), do Quociente de Eficiência Protéica (ganho de peso dividido pelo consumo de proteína), da Razão Protéica Líquida (ganho de peso corrigido dividido pelo consumo de proteína) e da Digestibilidade Verdadeira. Resultados: Os resultados demonstraram que quando o Champignon do Brasil foi utilizado como fonte exclusiva de proteína na dieta, os índices de qualidade protéica apresentaram-se baixos (Quociente de Eficiência Alimentar= 0,08, Quociente de Eficiência Protéica=0,92 e Razão Protéica Líquida=3,00), quando comparados com a dieta padrão caseína (Quociente de Eficiência Alimentar=0,30, Quociente de Eficiência Protéica=3,05 e Razão Protéica Líquida=4,21). Os índices obtidos para o grupo Agaricus mostraram-se comparáveis àqueles 536 | G.S. HENRIQUES et al. Revista de Nutrição Rev. Nutr., Campinas, 21(5):535-543, set./out., 2008 apresentados por alguns tipos de proteína vegetal e podem ser explicados por sua limitação em aminoácidos essenciais, notadamente a lisina e a leucina, respectivamente primeiro e segundo aminoácido limitante. Conclusão: Os dados apontam para a utilização da proteína do Agaricus brasiliensis como uma boa fonte para complementação protéica, quando combinada com outras culturas vegetais comuns na dieta típica brasileira

Expressão de proteína capsídica recombinante do Apple stem pitting virus e produção de antissoro policlonal.

O objetivo deste trabalho foi a expressão do gene da proteína capsidíca do ASPV em bactérias e a produção do antissoro policlonal contra a proteína expressada.Resumo