Farinha de resídues da filetagem de tilápia em rações para alevines de piauçu (Leporinus macrocephalus)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Diversidade genética de espécies de Xanthomonas patogênicas a citros baseada em genes avr e leucine protein

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Escolha dos níveis nutricionais na determinação do nível-ótimo e no ajuste de modelos estatísticos utilizados em ensaios dose-resposta

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Positioning and number of nutritional levels in dose-response trials to estimate the optimal-level and the adjustment of the models

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Digestibilidade e exigência de aminoácidos para rã-touro

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeitos do LRP6 e Frizzled6 na diferenciação endotelial de DPSC

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

LRP1 modulates APP trafficking and APP metabolism within compartments of the secretory pathway. Increased AICD generation is ineffective in nuclear translocation and transcriptional activation

LRP1 modulates APP trafficking and metabolism within compartments of the secretory pathway The amyloid precursor protein (APP) is the parent protein to the amyloid beta peptide (Abeta) and is a central player in Alzheimer’s disease (AD) pathology. Abeta liberation depends on APP cleavage by beta- and gamma-secretases. To date, only a unilateral view of APP processing exists, excluding other proteins, which might be transported together and/or processed dependent on each other by the secretases described above. The low density lipoprotein receptor related protein 1 (LRP1) was shown to function as such a mediator of APP processing at multiple steps. Newly synthesized LRP1 can interact with APP, implying an interaction between these two proteins early in the secretory pathway. Therefore, we wanted to investigate whether LRP1 can mediate APP trafficking along the secretory pathway, and, if so, whether it affects APP processing. Indeed, we demonstrate that APP trafficking is strongly influenced by LRP1 transport through the endoplasmic reticulum (ER) and Golgi compartments. LRP1-constructs with ER- and Golgi-retention motifs (LRP-CT KKAA, LRP-CT KKFF) had the capacity to retard APP trafficking at the respective steps in the secretory pathway. Here, we provide evidence that APP metabolism occurs in close conjunction with LRP1 trafficking, highlighting a new role of lipoprotein receptors in neurodegenerative diseases. Increased AICD generation is ineffective in nuclear translocation and transcriptional activity A sequence of amyloid precursor protein (APP) cleavages gives rise to the APP intracellular domain (AICD) together with amyloid beta peptide (Abeta) and/or p3 fragment. One of the environmental factors identified favouring the accumulation of AICD appears to be a rise in intracellular pH. This accumulation is a result of an abrogated cleavage event and does not extend to other secretase substrates. AICD can activate the transcription of artificially expressed constructs and many downstream gene targets have been discussed. Here we further identified the metabolism and subcellular localization of the constructs used in this well documented gene reporter assay. We also co-examined the mechanistic lead up to the AICD accumulation and explored possible significances for its increased expression. We found that most of the AICD generated under pH neutralized conditions is likely that cleaved from C83. Furthermore, the AICD surplus is not transcriptionally active but rather remains membrane tethered and free in the cytosol where it interacts with Fe65. However, Fe65 is still essential in AICD mediated transcriptional transactivation although its exact role in this set of events is unclear.

Progettazione del layout di un nuovo stabilimento produttivo: il caso Fiorini Industries S.r.l.

Progettazione del nuovo layout per un nuovo stabilimento produttivo, in cui il gruppo Fiorini Industries S.r.l., in un'ottica di espansione e miglioramento dei parametri tecnici e dei flussi aziendali, si insedia lasciando la sua storica sede produttiva. Questo bisogno nasce dall'impossibilità di espansione dello stabilimento odierno e dalla necessità di ridurre i costi di movimentazione dei materiali durante il ciclo di produzione. La tesi si colloca in questo contesto di necessità di verificare le prestazioni del lay-out attuale, valutandone le problematiche e le criticità al fine di potere identificare delle valide soluzioni di layout per la nuova sede produttiva dal punto di vista tecnico-economico e ambientale. Il CAPITOLO1 è volto ad inquadrare il problema affrontato nella situazione odierna; mettendo in evidenza la rilevanza del lay-out. Viene poi spiegato in maniera approfondita l'approccio di progettazione seguito. Al CAPITOLO 2 spetta la presentazione dell'azienda, della sua storia, della sua gamma di prodotti e dei suoi reparti di lavorazione. L'inquadramento aziendale è necessario al fine di capire le esigenze aziendali di cui si dovrà tenere conto durante lo svolgimento del lavoro. Nel CAPITOLO 3 si procede con l'analisi della situazione attuale, recuperando i dati di input necessari alla determinazione del mix produttivo offerto al mercato, quindi con l'analisi dei cicli produttivi, dei flussi e delle risorse impiegate per le movimentazioni. Il CAPITOLO 4 illustra il nuovo polo industriale del gruppo e presenta le alternative di lay-out individuate, che vengono analizzate dal punto di vista economico per individuare il possibile saving economico annuo rispetto alla situazione attuale. Per completezza di informazione vengono presentate anche aree che non vengono progettate tramite l'analisi tecnico economica (Blocco 2 e Uffici). Il capitolo termina con la scelta del nuovo layout per la nuova sede, dove vengono studiati i saving annui in base ai mezzi di movimentazioni attuali e quelli implementabili. L'ultima parte dell'elaborato valuta le emissioni di anidride carbonica dovute alle movimentazioni interne e valuta la possibile riduzione di emissioni di CO2 con il nuovo layout.

Market Prices and Food Aid Local and Regional Procurement and Distribution: A Multi-Country Analysis

To date, no research has rigorously addressed the concern that local and regional procurement (LRP) of food aid could affect food prices and food price volatility in food aid source and recipient countries. We assemble spatially and temporally disaggregated data and estimate the relationship between food prices and their volatility and local food aid procurement and distribution across seven countries for several commodities. In most cases, LRP activities have no statistically significant relationship with either local price levels or food price volatility. The few exceptions underscore the importance of market monitoring. (C) 2013 Elsevier Ltd. All rights reserved.

The Timeliness and Cost-Effectiveness of the Local and Regional Procurement of Food Aid

Local and regional procurement (LRP) of food aid is often claimed to lead to quicker and more cost-effective response. We generate timeliness and cost-effectiveness estimates by comparing US-funded LRP activities in nine countries against in-kind, transoceanic food aid shipments from the US to the same countries during the same timeframe. Procuring food locally or distributing cash or vouchers results in a time savings of nearly 14 weeks, a 62 percent gain. Cost-effectiveness varies significantly by commodity type. Procuring grains locally saved over 50 percent, on average, while local procurement of processed commodities was not always cost-effective. (C) 2013 Elsevier Ltd. All rights reserved.

Tradeoffs or Synergies? Assessing Local and Regional Food Aid Procurement through Case Studies in Burkina Faso and Guatemala

We compare the impacts across a range of criteria of local and regional procurement (LRP) relative to transoceanic shipment of food aid in Burkina Faso and Guatemala. We find that neither instrument dominates the other across all criteria in either country, although LRP commonly performs at least as well as transoceanic shipment with respect to timeliness, cost, market price impacts, satisfying recipients' preferences, food quality and safety, and in benefiting smallholder suppliers. LRP is plainly a valuable food assistance tool, but its advantages and disadvantages must be carefully weighed, compared, and prioritized depending on the context and program objectives. (C) 2013 Elsevier Ltd. All rights reserved.

CD34-related coexpression of MDR1 and BCRP indicates a clinically resistant phenotype in patients with acute myeloid leukemia (AML) of older age

Clinical resistance to chemotherapy in acute myeloid leukemia (AML) is associated with the expression of the multidrug resistance (MDR) proteins P-glycoprotein, encoded by the MDR1/ABCB1 gene, multidrug resistant-related protein (MRP/ABCC1), the lung resistance-related protein (LRP), or major vault protein (MVP), and the breast cancer resistance protein (BCRP/ABCG2). The clinical value of MDR1, MRP1, LRP/MVP, and BCRP messenger RNA (mRNA) expression was prospectively studied in 154 newly diagnosed AML patients >or=60 years who were treated in a multicenter, randomized phase 3 trial. Expression of MDR1 and BCRP showed a negative whereas MRP1 and LRP showed a positive correlation with high white blood cell count (respectively, p < 0.05, p < 0.001, p < 0.001 and p < 0.001). Higher BCRP mRNA was associated with secondary AML (p < 0.05). MDR1 and BCRP mRNA were highly significantly associated (p < 0.001), as were MRP1 and LRP mRNA (p < 0.001) expression. Univariate regression analyses revealed that CD34 expression, increasing MDR1 mRNA as well as MDR1/BCRP coexpression, were associated with a lower complete response (CR) rate and with worse event-free survival and overall survival. When adjusted for other prognostic actors, only CD34-related MDR1/BCRP coexpression remained significantly associated with a lower CR rate (p = 0.03), thereby identifying a clinically resistant subgroup of elderly AML patients.

Mechanistic insights from the crystal structures of a feast/famine regulatory protein from Mycobacterium tuberculosis H37Rv

Rv3291c gene from Mycobacterium tuberculosis codes for a transcriptional regulator belonging to the (leucine responsive regulatory protein/regulator of asparigine synthase C gene product) Lrp/AsnC-family. We have identified a novel effectorbinding site from crystal structures of the apo protein, complexes with a variety of amino acid effectors, X-ray based ligand screening and qualitative fluorescence spectroscopy experiments. The new effector site is in addition to the structural characterization of another distinct site in the protein conserved in the related AsnC-family of regulators. The structures reveal that the ligandbinding loops of two crystallographically ndependent subunits adopt different conformations to generate two distinct effector-binding sites. A change in the conformation of the binding site loop 100–106 in the B subunit is apparently necessary for octameric association and also allows the loop to interact with a bound ligand in the newly identified effector-binding site. There are four sites of each kind in the octamer and the protein preferentially binds to aromatic amino acids. While amino acids like Phe, Tyr and Trp exhibit binding to only one site, His exhibits binding to both sites. Binding of Phe is accompanied by a conformational change of 3.7A ° in the 75–83 loop, which is advantageously positioned to control formation of higher oligomers. Taken together, the present studies suggest an elegant control mechanism for global transcription regulation involving binding of ligands to the two sites, individually or collectively.

Mechanisms regulating the p120-catenin/Kaiso pathway

The Wnt pathways contribute to many processes in cancer and developmental biology, with β-catenin being a key canonical component. P120-catenin, which is structurally similar to β-catenin, regulates the expression of certain Wnt target genes, relieving repression conferred by the POZ/ zinc-finger transcription factor Kaiso. In my first project, employing Xenopus embryos and mammalian cell lines, I found that the degradation machinery of the canonical Wnt pathway modulates p120-catenin protein stability, especially p120 isoform-1, through mechanisms shared with b-catenin. Exogenous expression of destruction-complex components such as GSK3b or Axin promotes p120-catenin degradation, and consequently, is able to rescue developmental phenotypes resulting from p120 over-expression during early Xenopus embryonic development. Conversely, as predicted, the in vivo depletion of either Axin or GSK3b coordinately increased p120 and b-catenin levels, while p120 levels decreased upon LRP5/6 depletion, which are positive modulators in the canonical Wnt pathway. At the primary sequence level, I resolved conserved GSK3b phosphorylation sites in p120’s (isoform 1) amino-terminal region. Point-mutagenesis of these residues inhibited the association of destruction complex proteins including those involved in ubiquitination, resulting in p120-catenin stabilization. Importantly, we found that two additional p120-catenin family members, ARVCF-catenin and d-catenin, in common with b-catenin and p120, associate with Axin, and are degraded in Axin’s presence. Thus, by similar means, it appears that canonical Wnt signals coordinately modulate multiple catenin proteins having roles in development and conceivably disease states. In my second project, I found that the Dyrk1A kinase exhibits a positive effect upon p120-catenin levels. That is, unlike the negative regulator GSK3b kinase, a candidate screen revealed that Dyrk1A kinase enhances p120-catenin protein levels via increased half-life. Dyrk1A is encoded by a gene located within the trisomy of chromosome 21, which contributes to mental retardation in Down Syndrome patients. I found that Dyrk1A expression results in increased p120 protein levels, and that Dyrk1A specifically associates with p120 as opposed to other p120-catenin family members or b-catenin. Consistently, Dyrk1A depletion in mammalian cell lines and Xenopus embryos decreased p120-catenin levels. I further confirmed that Dyrk overexpression and knock-down modulates both Siamois and Wnt11 gene expression in the expected manner based upon the resulting latered levels of p120-catenin. I determined that Dyrk expression rescues Kaiso depletion effects (gastrulation failure; increased endogenous Wnt11 expression), and vice versa. I then identified a putative Dyrk phosphorylation region within the N-terminus of p120-catenin, which may also be responsible for Dyrk1A association. I went on to make a phosphomimic mutant, which when over-expressed, had the predicted enhanced capacity to positively modulate endogenous Wnt11 and Siamois expression, and thereby generate gastrulation defects. Given that Dyrk1A modulates Siamois expression through stabilization of p120-catenin, I further observed that ectopic expression of Dyrk can positively influence b-catenin’s capacity to generate ectopic dorsal axes when ventrally expressed in early Xenopus embryos. Future work will investigate how Dyrk1A modulates the Wnt signaling pathway through p120-catenin, and possibly begin to address how dysfunction of Dyrk1A with respect to p120-catenin might relate to aspects of Down syndrome. In summary, the second phase of my graduate work appears to have revealed a novel aspect of Dyrk1A/p120-catenin action in embryonic development, with a functional linkage to canonical Wnt signaling. What I have identified as a “Dyrk1A/p120-catenin/Kaiso pathway” may conceivably assist in our larger understanding of the impact of Dyrk1A dosage imbalance in Down syndrome.