Utilização da metodologia WGCNA na construção de redes gênicas e identificação de mecanismos envolvidos na resposta de vacas à mastite.

Tendo em vista a importância da predição das funções biológicas de genes ainda não caracterizados e da identificação de redes gênicas, esse trabalho teve por objetivo implantar uma metodologia baseada em co-expressão para análise dos dados gerados nos experimentos de microarranjos da Rede Genômica Animal. Nesse trabalho, dados de um experimento com microarranjos da Embrapa Gado de Leite, realizado para se identificar genes e mecanismos biológicos envolvidos na resposta de vacas leiteiras a infecção por microorganismos causadores da mastite foram utilizados

Identification of mechanisms involved in mastitis response by means of gene network building.

Mastitis, an inflammation of the mammary gland, is the most prevalent and costly production disease in dairy herds worldwide. It is caused generally by bacteria and accounts for a significant decrease in milk production and quality. One promising approach to reduce problems caused by mastitis, in addition to sanitary care, is the selection of animals resistant to disease and the incorporation of this trait into the herds. Therefore, studies to better understand the mechanisms involved in animal response to this disease are essential to the proposition of new advances in area. In this context, the aim of this study was to identify groups of genes involved in cow response to mastitis infection, through gene network building from microarray data. Gene expression data from the GeneChipâ Bovine Genome Array (Affymetrix) hybridization with milk somatic cells samples from Holstein-Zebu crossbreed dairy cows, obtained before (B) and 24 hrs after (A) artificial infection with Staphylococcus agalactiae, were analyzed using a network building methodology based on gene co-expression. We used WGCNA (Weighted Gene Co-expression Analysis), a systems biology method for describing the correlation patterns among genes across microarray samples, that can be used for finding clusters (modules) of highly correlated genes to identify modules of co-expressed genes, which may correspond to functionally related genes. By comparing two networks (between contrasting data sets), conserved and non-conserved modules can be identified. This strategy, named differential network analysis, aims to identify genes groups that are both differentially expressed and differentially connected, and changes in connectivity may correspond to large-scale rewiring, in response to environmental changes and/or physiologic perturbations. Two microarray data sets, B (n=5) and A (n=5), were preprocessed using affy and gcrma R/Bioconductor packages. A filter was applied, which resulted in the use of only those transcripts present in all samples. Gene co-expression networks were identified separately for each group (B and A), by the R/WGCNA package. Gene networks were compared between the two groups, and non-conserved modules were uncovered from a correlation test of the connectivity values. Our analysis identified a total of 17 modules of co-expressed genes, three of them, designed by the colors grey (n=35), blue (n=37) and turquoise (n=192), non-conserved between the groups. Using Blast2GO for enrichment analysis, we find the molecular function Protein Binding overrepresented in all three modules. However, in despite of the same molecular function, each one of the modules showed distinct characteristcs. Genes of grey module (BTG3, CD3E, MBD1, CHIC2, PLXNA3, MOCS3, NEIL1, VPS45, BCL2) were related to apoptosis and antigen recognition. Genes of turquoise module were enriched in inflammation mediators, including known mastitis marker genes (FGL1, GJA1, F2RL1, PTPRF, S100A2, TGFB2). The blue one uncovered genes involved in cell division and inflammatory response (CD97, MAD2L1, ZFP106, CDKN2C, LOC514364, NOP14, PCBD1, LOC100139798, AP1S1, EDN1, IL1B, ANXA11). Our study identified some mechanisms (represented by gene modules) that have changed in cows in early response to mastitis infection. Further analysis are being carried out, based on these results, to advance the understanding of animals response to the disease, which can lead to identify the candidate genes that could be used in breeding programs.X-MEETING 2011

First cases of combined full robotic partial nephrectomy and colorectal resections: Results and new perspectives

Background: Nowadays the robotic platform is widespread in general surgery, urology, and gynecology. Combined surgery may represent an alternative to sequential procedures and it allows the treatment, at the same time, of coexisting lesions; in this perspective, full-robotic multiorgan surgery is starting to gain interest from surgeons worldwide. Methods: Between April and June 2019, two patients presenting with synchronous colorectal and kidney cancers underwent, respectively, full-robotic right colectomy with right partial nephrectomy and anterior rectal resection with left partial nephrectomy. Surgeries were performed by both the general surgery and urology team. Results: No intraoperative complications were registered and the postoperative course was uneventful in both cases. Conclusions: Combined multiple organ surgery with full robotic technique is safe and offers oncological adequate results. A multi-team surgical pre-planning is mandatory to reduce invasiveness and operative time. To the best of our knowledge, these are the first reports of full robotic partial nephrectomy combined with colorectal procedures

CT imaging findings of abdominopelvic vascular compression syndromes: what the radiologist needs to know

Abdominopelvic vascular compression syndromes include a variety of uncommon conditions characterized by either extrinsic compression of blood vessels by adjacent anatomical structures (i.e., median arcuate ligament syndrome, nutcracker syndrome, May-Thurner syndrome) or compression of hollow viscera by adjacent vessels (i.e., superior mesenteric artery syndrome, ureteropelvic junction obstruction, ureteral vascular compression syndromes, portal biliopathy). These syndromes can be unexpectedly diagnosed even in asymptomatic patients and the predisposing anatomic conditions can be incidentally discovered on imaging examinations performed for other indications, or they can manifest with atypical abdominal symptoms and acute complications, which may lead to significant morbidity if unrecognized. Although computed tomography (CT) is an accurate noninvasive technique for their detection, the diagnosis remains challenging due to the uncommon clinical presentation and often overlooked imaging features. Dynamic imaging may be performed in order to evaluate patients with inconstant symptoms manifesting in a specific position. The purposes of this paper are to review the CT imaging findings of abdominopelvic vascular compression syndromes, correlating with anatomical variants and to provide key features for the noninvasive imaging diagnosis

CT angiography for the assessment of EVAR complications: a pictorial review

Endovascular aneurysm repair (EVAR) is a minimally invasive treatment proposed as an alternative to open repair in patients with abdominal aortic aneurysms. EVAR consists in a stent-graft placement within the aorta in order to exclude the aneurysm from arterial circulation and reduce the risk of rupture. Knowledge of the various types of devices is mandatory because some stents/grafts are more frequently associated with complications. CT angiography is the gold standard diagnostic technique for preprocedural planning and postprocedural surveillance. EVAR needs long-term follow-up due to the high rate of complications. Complications can be divided in endograft device-related and systemic complications. The purpose of this article is to review the CT imaging findings of EVAR complications and the key features for the diagnosis

Beam-induced damage on diffractive hard X-ray optics

Beam-induced damage on diffractive hard X-ray optics is studied by means of X-ray diffraction and scanning electron microscopy

Corrole Isomers: Intrinsic Gas-phase Shapes Via Traveling Wave Ion Mobility Mass Spectrometry And Dissociation Chemistries Via Tandem Mass Spectrometry.

Corrole and four of its isomers with subtle structural changes promoted by exchange of nitrogen and carbon atoms in the corrole ring have been studied by traveling wave ion mobility mass spectrometry and collision induced dissociation experiments. Significant differences in shapes and charge distributions for their protonated molecules were found to lead to contrasting gas phase mobilities, most particularly for corrorin, the most confused isomer. Accordingly, corrorin was predicted by B3LYP/6-31g(d,p) and collisional cross section calculations to display the most compact tri-dimensional structure, whereas NCC4 and corrole were found to be the most planar isomers. Better resolution between the corrole isomers was achieved using the more polarizable and massive CO(2) as the drift gas. Sequential losses of HF molecules were found to dominate the dissociation chemistry of the protonated molecules of these corrole isomers, but their unique structures caused contrasting labilities towards CID, whereas NCC4 showed a peculiar and structurally diagnostic loss of NH(3), allowing its prompt differentiation from the other isomers.108396-40

Proteome of the phytopathogen Xanthomonas citri subsp. citri: a global expression profile

<p>Abstract</p> <p>Background</p> <p>Citrus canker is a disease caused by <it>Xantomonas citri </it>subsp.<it>citri (Xac)</it>, and has emerged as one of the major threats to the worldwide citrus crop because it affects all commercial citrus varieties, decreases the production and quality of the fruits and can spread rapidly in citrus growing areas. In this work, the first proteome of <it>Xac </it>was analyzed using two methodologies, two-dimensional liquid chromatography (2D LC) and tandem mass spectrometry (MS/MS).</p> <p>Results</p> <p>In order to gain insight into the metabolism of <it>Xac</it>, cells were grown on two different media (<b>NB </b>- Nutrient Broth and <b>TSE </b>- Tryptone Sucrose broth enriched with glutamic acid), and proteins were proteolyzed with trypsin and examined by 2D LC-MS/MS. Approximately 39% of all predicted proteins by annotation of <it>Xac </it>were identified with their component peptides unambiguously assigned to tandem mass spectra. The proteins, about 1,100, were distributed in all annotated functional categories.</p> <p>Conclusions</p> <p>This is the first proteomic reference map for the most aggressive strain of <it>Xanthomonas </it>pathogen of all orange varieties. The compilation of metabolic pathways involved with bacterial growth showed that <it>Xac </it>expresses a complete central and intermediary metabolism, replication, transcription and translation machineries and regulation factors, distinct membrane transporters (ABC, MFS and pumps) and receptors (MCP, TonB dependent and metabolites acquisition), two-component systems (sensor and regulatory components) and response regulators. These data corroborate the growth curve <it>in vitro </it>and are the first reports indicating that many of these genome annotated genes are translated into operative in <it>Xac</it>. This proteomic analysis also provided information regarding the influence of culture medium on growth and protein expression of <it>Xac</it>.</p

C7orf59/LAMTOR4 phosphorylation and structural flexibility modulate ragulator assembly

Ragulator is a pentamer composed of p18, MP1, p14, C7orf59, and hepatitis B virus X-interacting protein (HBXIP; LAMTOR 1-5) which acts as a lysosomal scaffold of the Rag GTPases in the amino acid sensitive branch of TORC1 signaling. Here, we present the crystal structure of human HBXIP-C7orf59 dimer (LAMTOR 4/5) at 2.9 angstrom and identify a phosphorylation site on C7orf59 which modulates its interaction with p18. Additionally, we demonstrate the requirement of HBXIP-C7orf59 to stabilize p18 and allow further binding of MP1-p14. The structure of the dimer revealed an unfolded N terminus in C7orf59 (residues 1-15) which was shown to be essential for p18 binding. Full-length p18 does not interact stably with MP1-p14 in the absence of HBXIP-C7orf59, but deletion of p18 residues 108-161 rescues MP1-p14 binding. C7orf59 was phosphorylated by protein kinase A (PKA) in vitro and mutation of the conserved Ser67 residue to aspartate prevented phosphorylation and negatively affected the C7orf59 interaction with p18 both in cell culture and in vitro. C7orf59 Ser67 was phosphorylated in human embryonic kidney 293T cells. PKA activation with forskolin induced dissociation of p18 from C7orf59, which was prevented by the PKA inhibitor H-89. Our results highlight the essential role of HBXIP-C7orf59 dimer as a nucleator of pentameric Ragulator and support a sequential model of Ragulator assembly in which HBXIP-C7orf59 binds and stabilizes p18 which allows subsequent binding of MP1-p149915891602CNPQ - Conselho Nacional de Desenvolvimento Científico e TecnológicoFAPESP – Fundação de Amparo à Pesquisa Do Estado De São Paulo2014/12445-0; 2017/21455-7; 2014/17264-3190174/2012-