Ramos Mesonero, Alicia: Memoria de las presas de Franco

Review of: Ramos Mesonero, Alicia. Memoria de las presas de Franco. Madrid: Huerga y Fierro editores, 2012. 414 pp

Genetic factors involved in stroke susceptibility and in outcome at three months

Tese de doutoramento, Biologia (Genética), Universidade de Lisboa, Faculdade de Ciências, 2011Stroke is a significant cause of death and disability in developed countries. It is a multifactorial disease, resulting from the interplay between genes and well-known life-style/environmental risk factors. Numerous studies have attempted to identify the genetic risk factors predisposing to stroke, but few have investigated the genetic factors involved in stroke outcome. This work aimed at the identification of genes contributing to stroke and influencing patient’s outcome after three months. Four inflammatory genes (IL1B, IL6, MPO and TNF) and two genes involved in the nitric oxide metabolism (NOS1 and NOS3) were tested for association with stroke. The results suggest that the IL6 and MPO genes influence stroke susceptibility through independent effects and non-additive interactions. Furthermore, they provided novel evidence for the involvement of the NOS1 gene in stroke susceptibility. Several studies have shown the important impact of oxidative stress, inflammation, angiogenesis, neurogenesis, neurovascular damage and neurovascular remodeling for stroke-associated brain damage and/or stroke recovery. Association analyses were thus carried out to assess the role of candidate genes involved in inflammatory processes (IL1B, IL6, MPO and TNF) and oxidative stress (NOS1 and NOS3), as well as matrix metalloproteinase genes (MMP2 and MMP9) and growth factor genes (BDNF, FGF2 and VEGFA) in patient’s outcome at three months. MMP2 genetic variants were found associated with patient’s outcome, and the results also indicate that two epistatic interactions between the BDNF and FGF2 genes and between the FGF2 and VEGFA genes influence this phenotype. A genome-wide association study was performed in stroke outcome using DNA pooled samples to provide novel insights into the mechanisms involved in stroke recovery. The BBS9 and GLIS3 genes were found associated with patient’s outcome at three months. Taken together, these results suggest that stroke susceptibility and outcome are modulated by a combination of main gene effects and gene-gene interactions, independently of stroke risk factors and/or severity parameters, highlighting the complexity of mechanisms predisposing to stroke and influencing recovery afterwards.O Acidente Vascular Cerebral (AVC) é uma das principais causas de morte e incapacidade permanente nos países desenvolvidos, tendo importantes consequências económicas e sociais. O AVC é considerado uma doença complexa, que resulta de uma acção combinada entre genes e factores de risco ambientais ou de estilo de vida. Muitos estudos foram já levados a cabo com o intuito de identificar os factores genéticos de risco para o AVC, mas os resultados têm sido inconsistentes. A maioria desses estudos analisou o papel de genes candidatos; mais recentemente, foram também realizados estudos de associação ao nível de todo o genoma. Ao contrário da susceptibilidade, poucos estudos procuraram identificar quais os factores genéticos envolvidos na recuperação após um AVC. Este trabalho de doutoramento pretendeu assim identificar genes que contribuam para a susceptibilidade ao AVC e que influenciem o estado de incapacidade funcional (outcome) do doente ao fim de três meses de recuperação. Para isso, foram utilizadas duas estratégias distintas. A primeira estratégia consistiu na análise do papel de genes candidatos nos dois fenótipos referidos anteriormente, genes esses que tinham sido escolhidos com base na sua função, nos resultados de estudos de associação anteriores e/ou nos resultados de estudos com modelos animais de AVC, factores de risco ou condições médicas associadas. Foi ainda realizado um estudo de associação ao nível de todo o genoma com o objectivo de identificar factores genéticos que influenciem o outcome do doente. Nesta segunda estratégia não existia uma hipótese a priori relativamente ao papel de um determinado gene no fenótipo, correspondendo por isso a uma análise não enviesada. Vários estudos têm sugerido que a inflamação e o stress oxidativo desempenham um papel relevante na susceptibilidade ao AVC. Factores de risco já conhecidos, como aterosclerose, diabetes, obesidade e hipertensão, estão associados a um perfil inflamatório elevado. Além disso, é também conhecida a importância das enzimas sintase do óxido nítrico (NOS) para a aterosclerose e a regulação da pressão sanguínea. Atendendo a isso, foi testada a associação de quatro genes inflamatórios (IL1B, IL6, MPO e TNF) e de dois genes envolvidos no metabolismo do óxido nítrico (NOS1 e NOS3) com o risco de AVC. Foram encontradas associações de variantes genéticas nos genes inflamatórios IL6 e MPO com a doença, assim como de uma interacção epistática entre eles contribuindo para o risco. Isto sugere que os dois genes influenciam a susceptibilidade ao AVC através de efeitos independentes e de efeitos de interacção não aditivos. Os resultados mostram ainda uma associação entre o AVC e variantes genéticas no gene NOS1, que codifica uma das isoformas de NOS. A análise de genes candidatos sugere assim que efeitos independentes dos genes inflamatórios ou de stress oxidativo IL6, MPO e NOS1, e efeitos não aditivos resultantes de interacções entre os genes IL6 e MPO têm um impacto na susceptibilidade ao AVC. Estes resultados são compatíveis e reforçam as observações feitas em estudos in vitro e in vivo relativamente ao papel da inflamação e do stress oxidativo nesta doença. É igualmente reconhecido o importante impacto de stress oxidativo, inflamação, angiogénese, neurogénese, dano e remodelação neurovasculares na lesão cerebral associada ao AVC e/ou na recuperação funcional dos doentes. Assim, foram também levados a cabo estudos de associação para avaliar o papel de genes candidatos envolvidos em processos inflamatórios (IL1B, IL6, MPO e TNF) e de stress oxidativo (NOS1 e NOS3), assim como de genes das metaloproteinases da matriz (MMPs) (MMP2 and MMP9) e de genes de factores de crescimento (BDNF, FGF2 and VEGFA), no outcome do doente após três meses de recuperação. Relativamente ao gene MMP2, os resultados mostram a associação entre variantes genéticas deste gene e o outcome do doente após três meses de recuperação. Vários estudos anteriores demonstraram que a ruptura da barreira hemato-encefálica associada ao AVC está relacionada com a expressão e activação de MMPs, levando a hemorragia, edema e morte celular. No entanto, outros estudos sugerem que a actividade destas proteínas pode ser benéfica na angiogénese e na remodelação neurovascular em fases tardias de recuperação, o que poderá contribuir para a recuperação funcional do doente. Neste trabalho de doutoramento foram ainda identificadas duas interacções epistáticas entre os genes BDNF e FGF2 e entre os genes FGF2 e VEGFA em associação com o outcome do paciente. Estes três genes codificam factores de crescimento que são partilhados pelo sistema nervoso e vascular e que afectam a homeostasia e desenvolvimento dos dois sistemas. Os factores de crescimento têm importantes funções ao nível da angiogénese, neurogénese e protecção neuronal, influenciando o estado neurológico dos doentes e a recuperação dos mesmos após o AVC. Tendo em conta estes resultados, a análise de genes candidatos sugere que o gene MMP2 e interacções epistáticas entre os genes BDNF e FGF2, e entre os genes FGF2 e VEGFA têm um impacto no outcome do doente ao fim de três meses de recuperação. Estes resultados são também compatíveis com estudos in vitro e in vivo que tinham demonstrado previamente a importância das MMPs e dos factores de crescimento na recuperação após um AVC. Foi feito um rastreio genómico com o objectivo de identificar factores genéticos que influenciem o outcome do doente. Para isso foi testada a associação de mais de 250 mil polimorfismos, localizados ao longo de todo o genoma, com o outcome do doente. Uma das vantagens desta estratégia é a de possibilitar a descoberta de novos mecanismos envolvidos neste fenótipo. Para realizar este estudo de uma forma economicamente eficiente foram analisados conjuntos (pools) de amostras de DNA de doentes. Após a identificação dos marcadores mais importantes com base em quatro estratégias distintas, esses resultados foram validados por genotipagem individual. Este estudo permitiu a identificação de uma associação entre os genes BBS9 e GLIS3 e o outcome do doente. Sabe-se que pacientes com a síndrome Bardet-Biedl têm mutações no gene BBS9. A obesidade é uma das manifestações clínicas mais importantes desta síndrome e foi demonstrado que, após um AVC, os danos neurológicos e as lesões cerebrais são menores em ratinhos em regime de restrição calórica do que em ratinhos sem esta restrição, o que sugere que o excesso de calorias consumidas/obesidade poderá influenciar o outcome após AVC. Relativamente ao gene GLIS3, que codifica um factor de transcrição, tinham sido anteriormente reportadas associações deste gene com a diabetes. Esta doença é um factor de risco para o AVC e está também associada com maior risco de morte e incapacidade funcional de doentes quando o AVC ocorre. Apesar de não ser imediatamente perceptível qual o papel dos genes BBS9 e GLIS3 no outcome dos doentes, é possível que estes genes tenham uma influência indirecta nesse fenótipo através de um efeito na diabetes e obesidade. Em conclusão, os resultados obtidos durante este trabalho de doutoramento sugerem que tanto a susceptibilidade ao AVC como o outcome do doente são modulados por uma combinação de efeitos de genes independentes e de interacções entre genes. Isto indica que os mecanismos envolvidos na predisposição a esta doença e na recuperação posterior dos doentes poderão ser bastante complexos. É de salientar que, com este trabalho de doutoramento, o número de genes candidatos analisados, até ao momento, na área da genética do outcome praticamente duplicou. Além disso, pela primeira vez, foi realizado um estudo de associação ao nível de todo o genoma com o outcome do doente. No futuro, será desejável aumentar a dimensão da amostra, em especial dos doentes com informação sobre recuperação. Deverão, ainda, ser realizados estudos para identificar as variantes genéticas causais que estão na base das associações encontradas com a susceptibilidade ao AVC e com o outcome do doente

As mulheres à margem do direito medieval - transgressoras, infratoras e pecadoras

Com a presente Dissertação procurámos compreender que mulheres eram marginalizadas pelo Direito na Idade Média tardia portuguesa, e o porquê dessa marginalização. Para o efeito, procurámos aferir o padrão do feminino enquanto construção cultural e social medieva - a mulher honrada - no seu cotejo com a transgressora, segundo as categorias definidas pelo Direito. Quisemos, assim, determinar os comportamentos femininos passíveis de desvalor jurídico e de acarretarem uma punição pela transgressão. Sob o ponto de vista metodológico, a tipologia das infrações baseou-se no levantamento das diferentes fontes jurídico-documentais, com especial enfoque nos séculos XIV e XV, mormente nas Ordenações do Senhor Rey D. Affonso V, nas Ordenações d'el Rei D. Duarte e no Livro das Leis e Posturas, alicerçada em bibliografia especializada; Women at the margins of medieval law - transgressors, offenders and sinners Abstract With this Dissertation, we sought to understand which women were marginalized by Law in the late Portuguese Middle Ages, and the justification for this marginalization. For this purpose, we pursued to assess the feminine standard as a cultural and social construction - the honorable woman - when compared with the transgressor, according to the categories defined by Law. Thus, we sought to ascertain the feminine behaviors subject to legal devaluation and punishment for transgression. From the methodological point of view, the typology of the infractions was based on a survey of the different legal documentation sources, with special focus on the 14th and 15th centuries, notably the Ordenações do Senhor Rey D. Affonso V, the Ordenações d'el Rei D. Duarte and the Livro das Leis e Posturas, supported by specialized bibliography

Influence of Glutamine and Glutamate Supplementation in the Blood Levels of Horses

Background: The most abundant free amino acid in mammals is glutamine (GLN). Little research has focused on GLN supplementation for horses, but GLN levels in this species are known to decline after exercise and during lactation. Under physiological conditions, the body produces Gln in sufficient quantities for general metabolism, and a small part of this amino acid comes from dietary protein. Little research has so far focused on equine dietary supplementation with free glutamine or combined with other amino acids during catabolic states or in highly stressful situations. This research was conducted to evaluate the effects of equine dietary supplementation using a combination of glutamine and glutamate.  Materials, Methods & Results: The study involved four Arabian mares, not in training (~380 kg; ~12 years old) and four treatments (control, and inclusions of 1, 2 and 4% of GLN+GLU) in a Latin square model. A 7-day washout period was established between each phase. Fifty percent of the mares’ maintenance energy requirements came from concentrate and 50% from hay and grazing. The other 50% came from Tifton hay (Cynodon dactylon), which was supplied ad libitum. After 7 weeks of nutritional supplementation (once a day, in the morning). In the experimental model, the mares were distributed in a Latin square design comprised of four treatments: control (without inclusion) and inclusions of 1%, 2% and 4% of supplement (AminoGut®, Ajinomoto do Brazil), and four animals. Blood was collected in five stages (fasting, and 60, 120, 240 and 360 min after feeding) in each treatment. The blood samples were analyzed to determine GLN, GLU, urea, creatinine, uric acid, total plasma protein, hematocrit and glucose levels. Glutamine and Glutamate concentrations were analyzed using the enzymatic spectrophotometric method. The results were analyzed statistically using one- and two-way ANOVA and Tukey’s test with P set at 5%. The results indicated that GLN differed in both the group (P 0.05). Significant changes in GLN levels were also observed in the 4% inclusion treatment compared to all the treatments in the fasting phase and in the + 60 min and +240 min phases of the control group (P 0.05) during the period under analysis, and remained within the normal range for the species in their current stabling conditions. The mares presented no clinical problems nor did they change their feeding behavior during the supplementation period or on the days blood was collected.Discussion: Glutamine metabolism in horses has yet to be extensively studied.  However, it has been shown that, when supplied to horses in its free form, this amino acid causes Gln levels to rise rapidly within the first 90 min of the postprandial period.  This indicates that an extra amount of this amino acid may increase Gln blood levels despite intense degradation of enterocytes. This study found that supplementation with a combination of Gln+Glu can increase blood Gln levels after 6 h in the treatment involving 4% Gln+Glu included in the concentrate (P < 0.05) In conclusion, supplementation with GLN+GLU raised the mares’ GLN levels after 360 min when 4% of GLN+GLU was included in their diet. These results may be used to establish GLN supplementation models for horses.

Supplementation with nutraceuticals produces changes in working horse’s blood parameters but not in their body composition

Background:  :  :  : There is little scientific data about the effects of continuous supplementation with nutraceuticals over horse’s health and performance. Horse owners and veterinarians are still using supplements for horses without any kind of evaluations. At the Brazilian market is possible to buy hundreds of different products, but there is not private or state regulation over production and quality of these products. The absence of information about quality and efficacy of these products may contribute to mistakes in horse’s nutrition program. The objective of this research was to determine the effect of supplementation with a combination of nutraceuticals over some blood parameters (red cell count, total plasma proteins, free plasma glutamine and glutamate) and components of body composition in working horses. Materials, Methods & Results: There were 19 gelding horses, divided in two groups: nutraceutical (NUTR; n=10) and placebo (PLAC; n=9). Both groups received equivalent feeding regimens in regard to concentrate and grass, and did similar exercise program. NUTR group received their pellets with nutraceuticals, and PLAC group received a placebo during the morning meal. There is no visual difference between nutraceutical and placebo pellets. Body composition determination and blood collection were taken in three periods: pre-test, after 4 and 12 weeks of supplementation. Body composition was determined after evaluation of horse’s body mass by electronic scale associated with measurement of rump fat by ultrasound. Results showed that there were no significant changes in body composition during 12 weeks of supplementation in both groups, which had percentage of body fat around 13%. However, supplementation with nutraceutical produced significant changes (P7%) and free plasma glutamine (>20%) observed in NUTR group will contribute to horse’s performance since it is positive action on red blood indexes and plasma amino acid concentration, contributing to the well-being of this horse group. Increase in blood glutamine concentration observed in NUTR group without improvement of body composition was not expected because this amino acid is produced mainly by skeletal muscle. Increase in fat-free mass frequently is associated with improvement of blood glutamine concentration. Finally, initial elevation in few parameters in PLAC group at 4 weeks of supplementation probably was associated with improvement of general management practices. In conclusion, Supplementation with a nutraceutical combination, for 12 weeks, produces changes in the concentration of some blood parameters, but it didn’t modify the corporal composition. The elevation of those components can favor the transport of oxygen and nutrients in tissues leading to improvement in the horse performance

Glutamine and Glutamate Supplementation Increases the Levels of These Amino Acids in the Milk of Pasture-fed Mares

Background: L-Glutamine (Gln), the most abundant free alpha amino acid in the body, plays a major role in the transport of nitrogen and carbon between tissues, and is an important source of respiratory energy for intestinal and immune system cells. Mares lose lean body mass during lactation, when plasma and milk Gln levels change significantly. However, supplementation with Gln combined with other amino acids may not alter equine plasma Gln levels. The work reported here was designed to test the hypothesis that supplementation with a mixture of glutamine and glutamate (AminoGut) alters blood and milk free glutamine and glutamate levels in pasture-fed lactating mares.Materials, Methods & Results: This study involved 31 multiparous Quarter Horse mares, which were divided into three groups immediately postpartum, as follows: G-CON (n = 19); G-50 g supplemented with 50 g of Gln + Glu plus 200 g of concentrate (n = 6); and G-100 g, supplemented with 100 g of Gln + Glu plus 200 g of concentrate (n = 6). Blood and milk samples were collected on the day of parturition prior to supplementation, and monthly until weaning. The milk samples were used to analyze the Gln, Glu composition and levels, while the blood samples were used for further analysis of blood biomarkers. The results were analyzed by ANOVA and by Tukey’s test and the P value was set at 5%. The G-CON group showed a significant reduction of 11-35% in the mean blood glutamine levels from the first month postpartum and throughout lactation. In contrast, blood glutamine levels in groups G-50 g and G-100 g did not change significantly from parturition through 5 months of lactation. The supplemented groups showed no significant differences in blood variables such as protein, albumin, urea, creatinine, cholesterol, triglycerides and minerals. Free glutamine levels in milk did not change from parturition through the end of lactation in the G-CON group, but groups G-50 g and the G-100 g showed a marked rise in milk glutamine levels throughout the first three months of lactation (~3x), which remained high (~2x) until the foals were weaned (P > 0.05).Discussion: The results of this study indicate that Gln + Glu supplementation successfully increased Gln levels in mare milk in the first three months of lactation, and Glu levels in G-100 g in the first four months, without affecting the levels of these amino acids in the animals’ blood, which remained similar to data obtained at parturition. In fact, the Gln levels in both supplemented groups exceeded 1,000 mmol/mL throughout lactation, unlike those of the control group and of the samples obtained at parturition. Moreover, supplementation did not produce significant changes in blood biomarkers, including those pertaining to protein metabolism (urea, creatinine, uric acid, albumin and total proteins), indicating that the product used for supplementation did not interfere in these biomarkers, which remained within the normal physiological variations for the species. It was concluded that daily dietary supplementation with 50 g of a mixture of glutamine and glutamate produced an effect similar to supplementation with 100 g/day. Both supplementation protocols succeeded in raising glutamine levels in mare milk in the first three months of lactation, without interfering in blood biomarkers or milk composition. In view of the cost of the product, we recommend that the daily diet of mares during lactation be supplemented with 50 g of a mixture of Gln + Glu in order to produce the desired nutritional effects

Effects of L-Arginine Supplementation on Lactating Mares and the Development of Foals

Background: Most animal species are able to produce Arginine (Arg) under normal conditions. However, in some situations, its degradation can be higher than its production. For example, during a period of lactation or disease, there is an increase in the consumption of Arg. In this case, endogenous production is not enough for the animal’s demands. Indeed, Arg supplementation in animals has several benefits for the animal’s body, such as the increase of angiogenesis, improvements in immunity and the reproductive system, as well as the stimulation of lactogenesis. During the early phase of growth, a deficiency of Arg could cause a reduction in the growth rate and metabolic activity of animals. Therefore, this amino acid is considered essential in some phases of the life of animals. However, very few studies of the supplementation of this amino acid in horses have been carried out. The aim of the present study was to characterize the effects of supplementing lactating mares and their foals with Arg.Materials, Methods &amp; Results: Lactating mares (n = 10) were divided into two groups (control group: n = 3 / supplemented group: n = 7) and maintained exclusively under grazing. The supplemented group received 50 g of Arg during the lactation period. Samples of milk and blood from mares and blood from foals were collected at different phases of the lactation period. The following parameters were measured in milk: Glutamine (Gln); Glutamate (Glu); protein; fat; casein; lactose; urea and total solids. The following parameters were measured in blood: Gln; Glu; total plasmatic protein (TPP); albumin; urea; creatinine; uric acid; triglycerides; total cholesterol; calcium (C); phosphorous (P); magnesium (Mg) and ferrous (Fe). In addition, the biometric parameters of Wither Height (WH), Chest Perimeter (CP), Cannon Bone Circumference (CBC) and Fat Percentage (FP) of foals were obtained. A significant increase of Gln was observed in the milk in both groups (P &lt; 0.05). The highest concentration of Gln was detected in the third month of the lactation period in the supplemented group (~ 2.26 mmol/mL), and the control group (~ 1.91 mmol/mL) during the same period. Gln did not alter in the blood (P &gt; 0.05), although Glu was higher in the control group in the first month of the lactation period (~ 0.21 mmol/mL) (P &lt; 0.05). An increase in uric acid (~ 0.19 mmol/L) in both groups on the day of birth (P &lt; 0.05). In the supplemented group, increases in triglycerides (~ 0.60 mmol/L), Ca (~ 2.90 mmol/L) and Mg (~ 0.52 mmol/L) were observed in the first month of the lactation (P &lt; 0.05). At birth, foals exhibited high levels of urea (~ 4.67 mmol/L) and uric acid (~ 0.21 mmol/L), and low levels of P (~ 2.02 mmol/L) (P &lt; 0.05). The levels of Gln in the blood of foals remained between 0.50 and 0.70 mmol/mL throughout the lactation period (P &gt; 0.05). Even when they were added (Gln + Glu), no differences were observed (P &gt; 0.05). However, when the biometric parameters were analyzed, significant variations were detected in almost all characteristics (weight, WH, CP and CBC). In particular, the control group exhibited higher body mass and CP in the fifth month, when compared with the group of foals born from supplemented mares (P &lt; 0.05). The FP did not alter in either of the groups analyzed (P &gt; 0.05).Discussion: The results indicate that the supplementation of lactating mares with Arg produced few alterations in the parameters analyzed for both mares and foals. In addition, the supplementation did not produce side effects among the supplemented animals

A Protocol for Systematic Review

This work is financed by national funds through the FCT—Foundation for Science and Technology, I.P., under the project UIDB/04585/2020.Obstructive sleep apnea is a sleep disorder with a high prevalence in the world population. The mandibular advancement device is one of the options for treating obstructive sleep apnea. Neck computed tomography and drug-induced sleep endoscopy are complementary diagnostic tests that may help predict the effectiveness of mandibular advancement devices. This study aims to analyze the best method for predicting the effectiveness of mandibular advancement devices in the therapeutic approach to obstructive sleep apnea. PubMed, Embase, Cochrane Central Register of Controlled Trials (CENTRAL), and Web of Science Core Collection databases will be comprehensively searched. We will include randomized clinical trials, non-randomized prospective or retrospective clinical studies, case controls, cohort studies, and case series. Two authors will independently conduct data extraction and assess the literature quality of the studies. The analysis of the included literature will be conducted by Revman 5.3 software. The outcomes that will be analyzed are craniofacial characteristics, cephalometric assessments, site and type of obstruction of the upper airway, mean values of the apnea-hypopnea index, and SaO2 verified in the initial and follow-up polysomnography. This study will provide reliable, evidence-based support for the clinical application of mandibular advancement devices for obstructive sleep apnea.publishersversionpublishe