Molecular cloning and sequence analysis of hamster CENP-A cDNA

BACKGROUND: The centromere is a specialized locus that mediates chromosome movement during mitosis and meiosis. This chromosomal domain comprises a uniquely packaged form of heterochromatin that acts as a nucleus for the assembly of the kinetochore a trilaminar proteinaceous structure on the surface of each chromatid at the primary constriction. Kinetochores mediate interactions with the spindle fibers of the mitotic apparatus. Centromere protein A (CENP-A) is a histone H3-like protein specifically located to the inner plate of kinetochore at active centromeres. CENP-A works as a component of specialized nucleosomes at centromeres bound to arrays of repeat satellite DNA. RESULTS: We have cloned the hamster homologue of human and mouse CENP-A. The cDNA isolated was found to contain an open reading frame encoding a polypeptide consisting of 129 amino acid residues with a C-terminal histone fold domain highly homologous to those of CENP-A and H3 sequences previously released. However, significant sequence divergence was found at the N-terminal region of hamster CENP-A that is five and eleven residues shorter than those of mouse and human respectively. Further, a human serine 7 residue, a target site for Aurora B kinase phosphorylation involved in the mechanism of cytokinesis, was not found in the hamster protein. A human autoepitope at the N-terminal region of CENP-A described in autoinmune diseases is not conserved in the hamster protein. CONCLUSIONS: We have cloned the hamster cDNA for the centromeric protein CENP-A. Significant differences on protein sequence were found at the N-terminal tail of hamster CENP-A in comparison with that of human and mouse. Our results show a high degree of evolutionary divergence of kinetochore CENP-A proteins in mammals. This is related to the high diverse nucleotide repeat sequences found at the centromere DNA among species and support a current centromere model for kinetochore function and structural plasticity

Resurrecting the Regulatory Properties of the Ostreococcus tauri ADP-Glucose Pyrophosphorylase Large Subunit

ADP-glucose pyrophosphorylase (ADP-Glc PPase) catalyzes the first committed step for the synthesis of glycogen in cyanobacteria and starch in green algae and plants. The enzyme from cyanobacteria is homotetrameric (α4), while that from green algae and plants is heterotetrameric (α2β2). These ADP-Glc PPases are allosterically regulated by 3-phosphoglycerate (3PGA, activator) and inorganic orthophosphate (Pi, inhibitor). Previous studies on the cyanobacterial and plant enzymes showed that 3PGA binds to two highly conserved Lys residues located in the C-terminal domain. We observed that both Lys residues are present in the small (α) subunit of the Ostreococcus tauri enzyme; however, one of these Lys residues is replaced by Arg in the large (β) subunit. In this work, we obtained the K443R and R466K mutants of the O. tauri small and large subunits, respectively, and co-expressed them together or with their corresponding wild type counterparts. Our results show that restoring the Lys residue in the large subunit enhanced 3PGA affinity, whereas introduction of an Arg residue in the small subunit reduced 3PGA affinity of the heterotetramers. Inhibition kinetics also showed that heterotetramers containing the K443R small subunit mutant were less sensitive to Pi inhibition, but only minor changes were observed for those containing the R466K large subunit mutant, suggesting a leading role of the small subunit for Pi inhibition of the heterotetramer. We conclude that, during evolution, the ADP-Glc PPase large subunit from green algae and plants acquired mutations in its regulatory site. The rationale for this could have been to accommodate sensitivity to particular metabolic needs of the cell or tissue

A Cross-Sectional Study of Prisoners in Mexico City Comparing Prevalence of Transmissible Infections and Chronic Diseases with That in the General Population.

ObjectivesTo describe patterns of transmissible infections, chronic illnesses, socio-demographic characteristics and risk behaviors in Mexico City prisons, including in comparison to the general population, to identify those currently needing healthcare and inform policy.Materials and methodsA cross-sectional study among 17,000 prisoners at 4 Mexico City prisons (June to December 2010). Participation was voluntary, confidential and based on informed consent. Participants were tested for HIV, Hepatitis B & C, syphilis, hypertension, obesity, and, if at risk, glucose and cholesterol. A subset completed a questionnaire on socio-demographic characteristics and risk behaviors. Positive results were delivered with counseling and treatment or referral.Results76.8% (15,517/20,196) of men and 92.9% (1,779/1,914) of women participated. Complete data sets were available for 98.8%. The following prevalence data were established for transmissible infections: HIV 0.7%; syphilis: Anti-TP+/VDRL+ 2.0%; Hepatitis B: HBcAb 2.8%, HBsAg 0.15%; Anti-HCV 3.2%. Obesity: 9.5% men, 33.8% women. Compared with national age- and sex-matched data, the relative prevalence was greater for HIV and syphilis among women, HIV and Hepatitis C in men, and all infections in younger participants. Obesity prevalence was similar for women and lower among male participants. The prevalence of previously diagnosed diabetes and hypertension was lower. Questionnaire data (1,934 men, 520 women) demonstrated lower educational levels, increased smoking and substance use compared to national data. High levels of non-sterile tattooing, physical abuse and histories of sexual violence were found.ConclusionThe study identified that health screening is acceptable to Mexico City prisoners and feasible on a large-scale. It demonstrated higher prevalence of HIV and other infections compared to national data, though low rates compared to international data. Individual participants benefited from earlier diagnosis, treatment and support. The data collected will also enable the formulation of improved policy for this vulnerable group

Genetic characterization of domestic pigs in the core zone of swine production of Argentina

Argentina is a small player in the global pork market, contributing only 0.7% of the total production. With increasing global demand for meat, there is an opportunity for countries with an agricultural profile to grow their pork production. However, there is a need to understand the current state of the pork production sector in all aspects to inform decision-making. The aim of this study was to genetically characterize pig herds from different production strata in the primary region for pork production in the country. For this purpose, phylogenetic and genetic variability analyses were performed using the mitochondrial control region marker (n=95 pig samples). Moreover, genotyping of ryr1 and PRKAG3 genes (n=108 pig samples) were performed to evaluate the frequency of deleterious alleles for meat quality traits in the region. The results showed high levels of genetic variability in the pig herds (Hd= 0.840 ± 0.031 and π= 0.010 ± 0.001), with a creole sow and Iberian lineage standing out in the phylogeny. The genotyping of the ryr1 marker revealed the presence of the deleterious t allele in all analyzed strata. However, the RN-allele of the PRKAG3 gene was detected only in the two lower strata. This study represents the first analysis of the phylogenetic relationships among domestic pigs from Argentina and provides an initial assessment of genetic variability in the region. Additionally, the results present, for the first time, the frequency of deleterious alleles for pig production in the productive core area, demonstrating their prevalence.Fil: Figueroa, Carlos Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia del Noroeste de la Provincia de Buenos Aires. Universidad Nacional del Noroeste de la Provincia de Buenos Aires. Centro de Investigaciones y Transferencia del Noroeste de la Provincia de Buenos Aires; ArgentinaFil: Mac Allister, Matias Exequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia del Noroeste de la Provincia de Buenos Aires. Universidad Nacional del Noroeste de la Provincia de Buenos Aires. Centro de Investigaciones y Transferencia del Noroeste de la Provincia de Buenos Aires; ArgentinaFil: Acosta, Diana Belén. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro de Investigaciones y Transferencia del Noroeste de la Provincia de Buenos Aires. Universidad Nacional del Noroeste de la Provincia de Buenos Aires. Centro de Investigaciones y Transferencia del Noroeste de la Provincia de Buenos Aires; ArgentinaFil: Fernández, G. P.. Universidad Nacional del Noroeste de la Provincia de Buenos Aires. Centro de Bioinvestigaciones (Sede Pergamino); ArgentinaFil: Merino, M. L.. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas; Argentin

Dynamic scaling exponents of copper electrodeposits from scanning force microscopy imaging : Influence of a thiourea additive on the kinetics of roughening and brightening

Changes in the topography of Cu electrodeposits grown on polycrystalline Cu substrates at low constant current density from still aqueous concentrated CuSO4 + H2SO4 solutions, at 298 K, were studied by scanning force microscopy (SFM) at different scale lengths (L) from the nanometer level upward. The dynamic scaling theory applied to SFM images leads to exponents α = 0.87 ± 0.06 and β = 0.63 ± 0.08, which are consistent with an interface growing under an unstable regime. For similar conditions, the addition of 1,3-diethyl-2-thiourea reduces the average crystal size (〈ds〉) of electrodeposits leading to scaling exponents α = 0.86 ± 0.06 and β = 0.24 ± 0.05 for L s⟩ and a logarithmic dependence for the spatial and temporal evolution of the interface for L > 3 μm and t → 0. In an additive-free plating bath, the unstable growth regime appears to be originated by enhanced electrodeposition at protrusions due to curvature effects and further sustained by the electric and concentration fields built up around the growing deposit. The presence of the additive hinders the development of instabilities driving the evolution of the growing interface to that predicted by the Edwards-Wilkinson growth model on the asymptotic limit.Instituto de Investigaciones Fisicoquímicas Teóricas y AplicadasFacultad de Ciencias Exacta

A short-term data based water consumption prediction approach

A smart water network consists of a large number of devices that measure a wide range of parameters present in distribution networks in an automatic and continuous way. Among these data, you can find the flow, pressure, or totalizer measurements that, when processed with appropriate algorithms, allow for leakage detection at an early stage. These algorithms are mainly based on water demand forecasting. Different approaches for the prediction of water demand are available in the literature. Although they present successful results at different levels, they have two main drawbacks: the inclusion of several seasonalities is quite cumbersome, and the fitting horizons are not very large. With the aim of solving these problems, we present the application of pattern similarity-based techniques to the water demand forecasting problem. The use of these techniques removes the need to determine the annual seasonality and, at the same time, extends the horizon of prediction to 24 h. The algorithm has been tested in the context of a real project for the detection and location of leaks at an early stage by means of demand forecasting, and good results were obtained, which are also presented in this paper.publishersversionpublishe

Deformation heterogeneity study of a 6061-T6 aluminum alloy processed by equal channel angular pressing

Among the severe plastic deformation techniques, the equal channel angular pressing (ECAP) has drastically improved the mechanical properties of the processed alloys. However, information regarding friction phenomenon, which modifies the deformation at the surface and the heterogeneity microstrain state produced by the process itself, is still scarce. In the present work, the deformation heterogeneity and the friction effect, at the surface in the bulk material of the 6061-T6 aluminum alloy processed by ECAP, is presented and discussed. The residual stress (RS) measurements were performed by means of X-Ray diffraction. By means of synchrotron diffraction, volumetric sections of the ECAPed samples were characterized. Finite element analysis showed a good agreement with the experimentally obtained residual stress and microhardness mapping results. The study also showed that the highest deformation zones were located at the outer parts of the deformed samples (top and bottom), while the inner zone showed strain oscillations of up to 49±2 MPa.Peer ReviewedPostprint (author's final draft

Dynamic scaling exponents of copper electrodeposits from scanning force microscopy imaging : Influence of a thiourea additive on the kinetics of roughening and brightening

Changes in the topography of Cu electrodeposits grown on polycrystalline Cu substrates at low constant current density from still aqueous concentrated CuSO4 + H2SO4 solutions, at 298 K, were studied by scanning force microscopy (SFM) at different scale lengths (L) from the nanometer level upward. The dynamic scaling theory applied to SFM images leads to exponents α = 0.87 ± 0.06 and β = 0.63 ± 0.08, which are consistent with an interface growing under an unstable regime. For similar conditions, the addition of 1,3-diethyl-2-thiourea reduces the average crystal size (〈ds〉) of electrodeposits leading to scaling exponents α = 0.86 ± 0.06 and β = 0.24 ± 0.05 for L s⟩ and a logarithmic dependence for the spatial and temporal evolution of the interface for L > 3 μm and t → 0. In an additive-free plating bath, the unstable growth regime appears to be originated by enhanced electrodeposition at protrusions due to curvature effects and further sustained by the electric and concentration fields built up around the growing deposit. The presence of the additive hinders the development of instabilities driving the evolution of the growing interface to that predicted by the Edwards-Wilkinson growth model on the asymptotic limit.Instituto de Investigaciones Fisicoquímicas Teóricas y AplicadasFacultad de Ciencias Exacta

The Crystal Structure of Nitrosomonas Europaea Sucrose Synthase Reveals Critical Conformational Changes and Insights into the Sucrose Metabolism in Prokaryotes

In this paper we report the first crystal structure of a prokaryotic sucrose synthase from the non-photosynthetic bacterium Nitrosomonas europaea. The obtained structure was in an open form, whereas the only other available structure from the plant Arabidopsis thaliana was in a closed conformation. Comparative structural analysis revealed a “hinge-latch” combination, which is critical to transition between the open and closed forms of the enzyme. The N. europaea sucrose synthase shares the same fold as the GT-B family of the retaining glycosyltransferases. In addition, a triad of conserved homologous catalytic residues in the family showed to be functionally critical in the N. europaea sucrose synthase (Arg567, Lys572, Glu663). This implies that sucrose synthase shares not only a common origin with the GT-B family, but also a similar catalytic mechanism. The enzyme preferred transferring glucose from ADP-glucose rather than UDP-glucose like the eukaryotic counterparts. This predicts that these prokaryotic organisms have a different sucrose metabolic scenario from plants. Nucleotide preference determines where the glucose moiety is targeted after sucrose is degraded