A Chemical and Enzymatic Approach to Study Site-Specific Sumoylation.

A variety of cellular pathways are regulated by protein modifications with ubiquitin-family proteins. SUMO, the Small Ubiquitin-like MOdifier, is covalently attached to lysine on target proteins via a cascade reaction catalyzed by E1, E2, and E3 enzymes. A major barrier to understanding the diverse regulatory roles of SUMO has been a lack of suitable methods to identify protein sumoylation sites. Here we developed a mass-spectrometry (MS) based approach combining chemical and enzymatic modifications to identify sumoylation sites. We applied this method to analyze the auto-sumoylation of the E1 enzyme in vitro and compared it to the GG-remnant method using Smt3-I96R as a substrate. We further examined the effect of smt3-I96R mutation in vivo and performed a proteome-wide analysis of protein sumoylation sites in Saccharomyces cerevisiae. To validate these findings, we confirmed several sumoylation sites of Aos1 and Uba2 in vivo. Together, these results demonstrate that our chemical and enzymatic method for identifying protein sumoylation sites provides a useful tool and that a combination of methods allows a detailed analysis of protein sumoylation sites

Shotguns vs Lasers: Identifying barriers and facilitators to scaling-up plant molecular farming for high-value health products.

Plant molecular farming (PMF) is a convenient and cost-effective way to produce high-value recombinant proteins that can be used in the production of a range of health products, from pharmaceutical therapeutics to cosmetic products. New plant breeding techniques (NPBTs) provide a means to enhance PMF systems more quickly and with greater precision than ever before. However, the feasibility, regulatory standing and social acceptability of both PMF and NPBTs are in question. This paper explores the perceptions of key stakeholders on two European Union (EU) Horizon 2020 programmes-Pharma-Factory and Newcotiana-towards the barriers and facilitators of PMF and NPBTs in Europe. One-on-one qualitative interviews were undertaken with N = 20 individuals involved in one or both of the two projects at 16 institutions in seven countries (Belgium, France, Germany, Italy, Israel, Spain and the UK). The findings indicate that the current EU regulatory environment and the perception of the public towards biotechnology are seen as the main barriers to scaling-up PMF and NPBTs. Competition from existing systems and the lack of plant-specific regulations likewise present challenges for PMF developing beyond its current niche. However, respondents felt that the communication of the benefits and purpose of NPBT PMF could provide a platform for improving the social acceptance of genetic modification. The importance of the media in this process was highlighted. This article also uses the multi-level perspective to explore the ways in which NPBTs are being legitimated by interested parties and the systemic factors that have shaped and are continuing to shape the development of PMF in Europe

Light-dark cycle synchronization of circadian rhythm in blind primates

BACKGROUND: Recently, several papers have shown that a small subset of retinal ganglion cells (RGCs), which project to the suprachiasmatic nucleus (SCN) and contain a new photopigment called melanopsin, are the photoreceptors involved in light-dark entrainment in rodents. In our primate colony, we found a couple of common marmosets (Callithrix jacchus) that had developed progressive and spontaneous visual deficiency, most likely because of retinal degeneration of cones and/or rods. In this study, we evaluated the photoresponsiveness of the circadian system of these blind marmosets. METHODS: Two blind and two normal marmosets were kept in cages with a controlled light-dark cycle (LD) to study photoentrainment, masking, and phase response to a dark pulse. RESULTS: Blind marmosets were entrained with the new LD cycle when light onsets were delayed and advanced by 6 hours. In constant light conditions, blind marmosets free-ran with a period of 23.2 hours, while normal animals free-ran with a period of 23.6 hours. All marmosets responded to dark pulses in the early subjective day with phase delays and with phase advances in the late subjective day. CONCLUSION: Our results demonstrate that light can synchronize circadian rhythms of blind marmosets and consequently, that this species could be a good primate model for circadian photoreception studies

A ADOLESCENTE E O FILHO DESNUTRIDO: CUIDADO CULTURAL

This study approaches the adolescent mothers of undernourished child family context being used the ethnographic method to contemplate on the cultural care accomplished by those mothers. It is noticed that the adolescent mothers accept of relatives and neighbors what is better for the care with the son. It is concluded by the need of an educational work that contextualize the care with undernourished child.Aborda o contexto cultural familiar da adolescente, mãe de criança desnutrida, utilizando-se o método etnográfico para refletir sobre o cuidado dessas mães para com seus filhos. Percebe-se que as mães adolescentes aceitam de familiares e vizinhos orientação sobre o que é melhor para o cuidado do filho. Concluise pela necessidade de um trabalho educativo que contextualize o cuidado da criança desnutrida em seus aspectos socio-culturais

d=3 Bosonic Vector Models Coupled to Chern-Simons Gauge Theories

We study three dimensional O(N)_k and U(N)_k Chern-Simons theories coupled to a scalar field in the fundamental representation, in the large N limit. For infinite k this is just the singlet sector of the O(N) (U(N)) vector model, which is conjectured to be dual to Vasiliev's higher spin gravity theory on AdS_4. For large k and N we obtain a parity-breaking deformation of this theory, controlled by the 't Hooft coupling lambda = 4 \pi N / k. For infinite N we argue (and show explicitly at two-loop order) that the theories with finite lambda are conformally invariant, and also have an exactly marginal (\phi^2)^3 deformation. For large but finite N and small 't Hooft coupling lambda, we show that there is still a line of fixed points parameterized by the 't Hooft coupling lambda. We show that, at infinite N, the interacting non-parity-invariant theory with finite lambda has the same spectrum of primary operators as the free theory, consisting of an infinite tower of conserved higher-spin currents and a scalar operator with scaling dimension \Delta=1; however, the correlation functions of these operators do depend on lambda. Our results suggest that there should exist a family of higher spin gravity theories, parameterized by lambda, and continuously connected to Vasiliev's theory. For finite N the higher spin currents are not conserved.Comment: 34 pages, 29 figures. v2: added reference

The ρ(1S,2S)\rho(1S,2S), ψ(1S,2S)\psi(1S,2S), Υ(1S,2S)\Upsilon(1S,2S) and ψt(1S,2S)\psi_t(1S,2S) mesons in a double pole QCD Sum Rule

We use the method of double pole QCD sum rule which is basically a fit with two exponentials of the correlation function, where we can extract the masses and decay constants of mesons as a function of the Borel mass. We apply this method to study the mesons: $\rho(1S,2S)$, $\psi(1S,2S)$, $\Upsilon(1S,2S)$ and $\psi_t(1S,2S)$. We also present predictions for the toponiuns masses $\psi_t(1S,2S)$ of m(1S)=357 GeV and m(2S)=374 GeV.Comment: 14 pages, 11 figures in Braz J Phys (2016

Automatic Detection of Malignant Masses in Digital Mammograms Based on a MCET-HHO Approach

Digital image processing techniques have become an important process within medical images. These techniques allow the improvement of the images in order to facilitate their interpretation for specialists. Within these are the segmentation methods, which help to divide the images by regions based on different approaches, in order to identify details that may be complex to distinguish initially. In this work, it is proposed the implementation of a multilevel threshold segmentation technique applied to mammography images, based on the Harris Hawks Optimization (HHO) algorithm, in order to identify regions of interest (ROIs) that contain malignant masses. The method of minimum cross entropy thresholding (MCET) is used to select the optimal threshold values for the segmentation. For the development of this work, four mammography images were used (all with presence of a malignant tumor), in their two views, craniocaudal (CC) and mediolateral oblique (MLO), obtained from the Digital Database for Screening Mammography (DDSM). Finally, the ROIs calculated were compared with the original ROIs of the database through a series of metrics, to evaluate the behavior of the algorithm. According to the results obtained, where it is shown that the agreement between the original ROIs and the calculated ROIs is significantly high, it is possible to conclude that the proposal of the MCET-HHO algorithm allows the automatic identification of ROIs containing malignant tumors in mammography images with significant accuracy.Digital image processing techniques have become an important process within medical images. These techniques allow the improvement of the images in order to facilitate their interpretation for specialists. Within these are the segmentation methods, which help to divide the images by regions based on different approaches, in order to identify details that may be complex to distinguish initially. In this work, it is proposed the implementation of a multilevel threshold segmentation technique applied to mammography images, based on the Harris Hawks Optimization (HHO) algorithm, in order to identify regions of interest (ROIs) that contain malignant masses. The method of minimum cross entropy thresholding (MCET) is used to select the optimal threshold values for the segmentation. For the development of this work, four mammography images were used (all with presence of a malignant tumor), in their two views, craniocaudal (CC) and mediolateral oblique (MLO), obtained from the Digital Database for Screening Mammography (DDSM). Finally, the ROIs calculated were compared with the original ROIs of the database through a series of metrics, to evaluate the behavior of the algorithm. According to the results obtained, where it is shown that the agreement between the original ROIs and the calculated ROIs is significantly high, it is possible to conclude that the proposal of the MCET-HHO algorithm allows the automatic identification of ROIs containing malignant tumors in mammography images with significant accuracy