Comparative RNAseq analysis of backfat tissue from local pig breeds.

Alentejano (AL) and Bísaro (BI) are the main local pig breeds in Portugal, but have no information comparing their transcriptomic activity. AL belongs to the Iberian branch, presenting lower growth rates, precociously high adipogenic activity and higher levels of unsaturated fatty acids (FAs) while BI pig is from the Celtic group, sharing ancestors with higher growth rate and leaner commercial breeds. This work intended to explore the genome function of AL and BI to better understand the underlying physiological mechanisms associated with body fat accretion, lipid composition and meat quality. Dorsal subcutaneous fat (DSF) samples were collected from AL and BI fattening pigs, with ~150kg BW at slaughter. Total RNA was obtained and sequenced for transcriptome analysis. Bioinformatic analyses using three different tools (Cufflinks, EdgeR and DESeq2) were performed. A total of 367, 137 and 155 differentially expressed genes (DEGs) (q-value0.8) were found using the Cufflinks, EdgeR and DESeq2 pipelines, respectively, between AL and BI DSF samples. EdgeR and DESeq2 shared a total 121 DEGs (~71% overlap) while Cufflinks showed divergent results (2.7% overlap with EdgeR and 5.5% with DESeq2). A functional enrichment analysis of the candidate DEGs was performed using Ingenuity Pathway Analysis. Synthesis of lipid, depletion of glycogen, mass of organism and accumulation of oleic acid were revealed as main involved functions (p-value<0.05) though no directional activation state was observed (-2<Zscore<2). Potential upstream regulators that explain the obtained results such as TCF7L2 and RIPK2 were predicted to be activated and inhibited in AL, respectively. Moreover, 4 causal networks with RIT2, KL, FLCN and RIPK2 as master regulators were inhibited in AL while another with PPARGC1B was activated. These results present the first high-throughput transcriptomic data involving these local breeds and can help explain the metabolic differences that occur in the adipose tissue and shed light into specific meat quality traits

CASA-Mot technology: how results are affected by the frame rate and counting chamber

For over 30 years, CASA-Mot technology has been used for kinematic analysis of sperm motility in different mammalian species, but insufficient attention has been paid to the technical limitations of commercial computer-aided sperm analysis (CASA) systems. Counting chamber type and frame rate are two of the most important aspects to be taken into account. Counting chambers can be disposable or reusable, with different depths. In human semen analysis, reusable chambers with a depth of 10 mm are the most frequently used, whereas for most farm animal species it is more common to use disposable chambers with a depth of 20 mm. The frame rate was previously limited by the hardware, although changes in the number of images collected could lead to significant variations in some kinematic parameters, mainly in curvilinear velocity (VCL). A frame rate of 60 frames s(-1) is widely considered to be the minimum necessary for satisfactory results. However, the frame rate is species specific and must be defined in each experimental condition. In conclusion, we show that the optimal combination of frame rate and counting chamber type and depth should be defined for each species and experimental condition in order to obtain reliable results

Quantitative and qualitative analysis of psychosocial factors affecting women’s entrepreneurship

This work aims to clarify the psychosocial variables that lead women to undertake and those that prevent them from doing so. Two studies were conducted using a mixed methodology to compensate for the inherent weaknesses of using each approach. The first study was based on the collection of quantitative data using the GloPEW questionnaire with a sample of 296 people. The second study, of a qualitative nature, was carried out through focus groups with a sample of 26 people. The results show that self-efficacy and emotional intelligence are the main factors to develop to promote entrepreneurship among women. Although the data show statistical strength, it seems necessary to expand the sample and incorporate more profiles of female entrepreneurs, for example, with different levels of training, given the complexity and variety of intervening factors.info:eu-repo/semantics/publishedVersio

Evaluation of structural intervention in the Quartel das Esquadras, Almeida (Portugal)

The Quartel das Esquadras is an 18th century infantry barrack located within the limits of the bulwarked fortress of Almeida, in Portugal. An ongoing adaptive reuse project of the building aims to recover its full capacity and also its importance to the fortified village of Almeida, by implementing a variety of new uses. While adaptive reuse projects of historical buildings are a very recommendable solution for maintaining those constructions whose use has become obsolete, these projects demand careful study because they may introduce changes in the structural behaviour of the buildings to preserve. The architectural intervention results in the need of the structure to withstand new and diverse imposed loads, as well as inserting passages and communication systems to enable a proper contemporary use of the building, fulfilling architectural requirements and current building codes. As the current configuration of the structure will be altered, the main objective of this paper is to evaluate the suitability of some of the proposed structural interventions. Such evaluation requires a good understanding of the current state of the structure and the alterations proposed by the project. The current condition has been characterized following a multidisciplinary approach comprising historical research, visual inspection, non-destructive testing and structural analysis. The alterations proposed by the project have been described, identifying possible sources of major structural problems. A portion of the building particularly affected by the alterations has been selected to carry out structural analysis. A comparative safety assessment of the selected area in both current and altered condition has been done through finite element modelling and nonlinear static analysis, resulting in an identification of the weaker points against the new implemented loads and alterations. Finally, proposals for the implementation of the studied intervention, as well as recommendations for future research and analyses, have been given.This work was financed by FEDER funds through the Competitiveness Factors Operational Programme - COMPETE and by national funds through FCT - Foundation for Science and Technology within the scope of the project POCI-01-0145-FEDER-007633

Transcriptomic profiling of skeletal muscle reveals candidate genes influencing muscle growth and associated lipid composition in Portuguese local pig breeds

Gene expression is one of the main factors to influence meat quality by modulating fatty acid metabolism, composition, and deposition rates in muscle tissue. This study aimed to explore the transcriptomics of the Longissimus lumborum muscle in two local pig breeds with distinct genetic background using next-generation sequencing technology and Real-Time qPCR. RNA-seq yielded 49 differentially expressed genes between breeds, 34 overexpressed in the Alentejano (AL) and 15 in the Bísaro (BI) breed. Specific slow type myosin heavy chain components were associated with AL (MYH7) and BI (MYH3) pigs, while an overexpression of MAP3K14 in AL may be associated with their lower loin proportion, induced insulin resistance, and increased inflammatory response via NFkB activation. Overexpression of RUFY1 in AL pigs may explain the higher intramuscular (IMF) content via higher GLUT4 recruitment and consequently higher glucose uptake that can be stored as fat. Several candidate genes for lipid metabolism, excluded in the RNA-seq analysis due to low counts, such as ACLY, ADIPOQ, ELOVL6, LEP and ME1 were identified by qPCR as main gene factors defining the processes that influence meat composition and quality. These results agree with the fatter profile of the AL pig breed and adiponectin resistance can be postulated as responsible for the overexpression of MAP3K140s coding product NIK, failing to restore insulin sensitivity

Comparative Transcriptomic Analysis of Subcutaneous Adipose Tissue from Local Pig Breeds

When compared to modern lean-type breeds, Portuguese local Alentejano (AL) and Bísaro (BI) pig breeds present a high potential for subcutaneous and intramuscular fat (IMF) deposition which contributes for better meat quality. The aim of this work was to explore the genome function to better understand the underlying physiological mechanisms associated with body fat accretion. Dorsal subcutaneous fat samples were collected at slaughter from adult animals (n = 4 for each breed) with ~150 kg body weight. Total RNA was obtained and sequenced for transcriptome analysis using DESeq2. A total of 458 di erentially expressed (DE) genes (q-value < 0.05) were identified, with 263 overexpressed in AL and 195 in BI. Key genes involved in de novo fatty acid biosynthesis, elongation and desaturation were upregulated in AL such as ACLY, FASN, ME1, ELOVL6 and SCD. A functional enrichment analysis of the DE genes was performed using Ingenuity Pathway Analysis. Cholesterol synthesis is suggested to be higher in AL via SREBF2, SCAP and PPARG, while lipolytic activity may be more active in BI through GH and AMPK signalling. Increased signalling of CD40 together with the predicted activation of INSIG1 and INSIG2 in BI suggests that this breed is more sensitive to insulin whereas the AL is less sensitive like the Iberian breed

Evolution of crystalline orientations in the production of ferritic stainless steel

Ferritic stainless steel EN 1.4016 is used in a wide range of applications, the most common ones related to sheet forming. Several problems in the post-processing of these steels relates to their texture and anisotropy. Therefore, it is necessary to know the mechanisms of texture formation in the subsequent stages of metal manufacturing processes. EBSD has been demonstrated as a successful characterisation technique for this purpose. It is known that during re-crystallisation of Fe-Cr steels, deviations from the desired.-fibre texture promote a decrease of deep drawability. Additionally, a-fibre damages formability. Subsequent cold rolling and annealing can enhance the deep drawing properties of the steel sheet. In this research, a standard sample and a modified one with optimised settings as regard to chemical composition and manufacturing process, to improve the formability properties, are characterised. To analyse the preferred orientation and the type of main fibre present in the material, ODF and Aztec Reclassify Phase, to calculate the content of martensite, were used

Short- and Long-Term Prognosis of Patients With Takotsubo Syndrome Based on Different Triggers: Importance of the Physical Nature

Background Takotsubo syndrome (TTS) is an acute reversible heart condition initially believed to represent a benign pathology attributable to its self-limiting clinical course; however, little is known about its prognosis based on different triggers. This study compared short- and long-term outcomes between TTS based on different triggers, focusing on various physical triggering events. Methods and Results We analyzed patients with a definitive TTS diagnosis recruited for the Spanish National Registry on TTS (RETAKO [Registry on Takotsubo Syndrome]). Short- and long-term outcomes were compared between different groups according to triggering factors. A total of 939 patients were included. An emotional trigger was detected in 340 patients (36.2%), a physical trigger in 293 patients (31.2%), and none could be identified in 306 patients (32.6%). The main physical triggers observed were infections (30.7%), followed by surgical procedures (22.5%), physical activities (18.4%), episodes of severe hypoxia (18.4%), and neurological events (9.9%). TTS triggered by physical factors showed higher mortality in the short and long term, and within this group, patients whose physical trigger was hypoxia were those who had a worse prognosis, in addition to being triggered by physical factors, including age >70 years, diabetes mellitus, left ventricular eyection fraction <30% and shock on admission, and increased long-term mortality risk. Conclusions TTS triggered by physical factors could present a worse prognosis in terms of mortality. Under the TTS label, there could be as yet undiscovered very different clinical profiles, whose differentiation could lead to individual better management, and therefore the perception of TTS as having a benign prognosis should be generally ruled out

Predictive model of pheochromocytoma based on the imaging features of the adrenal tumours

The purpose of our study was to develop a predictive model to rule out pheochromocytoma among adrenal tumours, based on unenhanced computed tomography (CT) and/or magnetic resonance imaging (MRI) features. We performed a retrospective multicentre study of 1131 patients presenting with adrenal lesions including 163 subjects with histological confirmation of pheochromocytoma (PHEO), and 968 patients showing no clinical suspicion of pheochromocytoma in whom plasma and/or urinary metanephrines and/or catecholamines were within reference ranges (non-PHEO). We found that tumour size was significantly larger in PHEO than non-PHEO lesions (44.3 +/- 33.2 versus 20.6 +/- 9.2 mm respectively; P < 0.001). Mean unenhanced CT attenuation was higher in PHEO (52.4 +/- 43.1 versus 4.7 +/- 17.9HU; P < 0.001). High lipid content in CT was more frequent among non-PHEO (83.6% versus 3.8% respectively; P < 0.001); and this feature alone had 83.6% sensitivity and 96.2% specificity to rule out pheochromocytoma with an area under the receiver operating characteristics curve (AUC-ROC) of 0.899. The combination of high lipid content and tumour size improved the diagnostic accuracy (AUC-ROC 0.961, sensitivity 88.1% and specificity 92.3%). The probability of having a pheochromocytoma was 0.1% for adrenal lesions smaller than 20 mm showing high lipid content in CT. Ninety percent of non-PHEO presented loss of signal in the out of phase MRI sequence compared to 39.0% of PHEO (P < 0.001), but the specificity of this feature for the diagnosis of non-PHEO lesions low. In conclusion, our study suggests that sparing biochemical screening for pheochromocytoma might be reasonable in patients with adrenal lesions smaller than 20 mm showing high lipid content in the CT scan, if there are no typical signs and symptoms of pheochromocytoma