Interaction with calmodulin is important for the secretion of thimet oligopeptidase following stimulation

Thimet oligopeptidase (EC 3.4.24.15; EP24.15) was originally described as a neuropeptide-metabolizing enzyme, highly expressed in the brain, kidneys and neuroendocrine tissue. EP24.15 lacks a typical signal peptide sequence for entry into the secretory pathway and is secreted by cells via an unconventional and unknown mechanism. In this study, we identified a novel calcium-dependent interaction between EP24.15 and calmodulin, which is important for the stimulated, but not constitutive, secretion of EP24.15. We demonstrated that, in vitro, EP24.15 and calmodulin physically interact only in the presence of Ca(2+), with an estimated K(d) value of 0.52 mu m. Confocal microscopy confirmed that EP24.15 colocalizes with calmodulin in the cytosol of resting HEK293 cells. This colocalization markedly increases when cells are treated with either the calcium ionophore A23187 or the protein kinase A activator forskolin. Overexpression of calmodulin in HEK293 cells is sufficient to greatly increase the A23187-stimulated secretion of EP24.15, which can be inhibited by the calmodulin inhibitor calmidazolium. The specific inhibition of protein kinase A with KT5720 reduces the A23187-stimulated secretion of EP24.15 and inhibits the synergistic effects of forskolin with A23187. Treatment with calmidazolium and KT5720 nearly abolishes the stimulatory effects of A23187 on EP24.15 secretion. Together, these data suggest that the interaction between EP24.15 and calmodulin is regulated within cells and is important for the stimulated secretion of EP24.15 from HEK293 cells.

Peptidomic Analysis of Human Cell Lines

Peptides have been proposed to function in intracellular signaling within the cytosol. Although cytosolic peptides are considered to be highly unstable, a large number of peptides have been detected in mouse brain and other biological samples. In the present study, we evaluated the peptidome of three diverse cell lines: SH-SY5Y, MCF7, and HEIC293 cells. A comparison of the peptidomes revealed considerable overlap in the identity of the peptides found in each cell line. The majority of the observed peptides are not derived from the most abundant or least stable proteins in the cell, and approximately half of the cellular peptides correspond to the N- or C- termini of the precursor proteins. Cleavage site analysis revealed a preference for hydrophobic residues in the PI position. Quantitative peptidomic analysis indicated that the levels of most cellular peptides are not altered in response to elevated intracellular calcium, suggesting that calpain is not responsible for their production. The similarity of the peptidomes of the three cell lines and the lack of correlation with the predicted cellular degradome implies the selective formation or retention of these peptides, consistent with the hypothesis that they are functional in the cells.

Differential Antitumor Effects of IgG and IgM Monoclonal Antibodies and Their Synthetic Complementarity-Determining Regions Directed to New Targets of B16F10-Nex2 Melanoma Cells

Malignant melanoma has increased incidence worldwide and causes most skin cancer-related deaths. A few cell surface antigens that can be targets of antitumor immunotherapy have been characterized in melanoma. This is an expanding field because of the ineffectiveness of conventional cancer therapy for the metastatic form of melanoma. In the present work, antimelanoma monoclonal antibodies (mAbs) were raised against B16F10 cells (subclone Nex4, grown in murine serum), with novel specificities and antitumor effects in vitro and in vivo. MAb A4 (IgG2ak) recognizes a surface antigen on B16F10-Nex2 cells identified as protocadherin beta(13). It is cytotoxic in vitro and in vivo to B16F10-Nex2 cells as well as in vitro to human melanoma cell lines. MAb A4M (IgM) strongly reacted with nuclei of permeabilized murine tumor cells, recognizing histone 1. Although it is not cytotoxic in vitro, similarly with mAb A4, mAb A4M significantly reduced the number of lung nodules in mice challenged intravenously with B16F10-Nex2 cells. The V(H) CDR3 peptide from mAb A4 and V(L) CDR1 and CDR2 from mAb A4M showed significant cytotoxic activities in vitro, leading tumor cells to apoptosis. A cyclic peptide representing A4 CDR H3 competed with mAb A4 for binding to melanoma cells. MAb A4M CDRs L1 and L2 in addition to the antitumor effect also inhibited angiogenesis of human umbilical vein endothelial cells in vitro. As shown in the present work, mAbs A4 and A4M and selected CDR peptides are strong candidates to be developed as drugs for antitumor therapy for invasive melanoma.

Naturally Acquired Antibodies to Plasmodium vivax Duffy Binding Protein (DBP) in Rural Brazilian Amazon

Duffy binding protein (DBP), a leading malaria vaccine candidate, plays a critical role ill Plasmodium vivax erythrocyte invasion. Sixty-eight of 366 (18.6%) subjects had IgG anti-DBP antibodies by enzyme-linked immunosorbent assay (ELISA) in a community-based cross-sectional survey ill the Brazilian Amazon Basin. Despite Continuous exposure to low-level malaria transmission, the overall seroprevalence decreased to 9.0% when the Population was reexamined 12 months later. Antibodies from 16 of 50 (360%) Subjects who were ELISA-positive at the baseline were able to inhibit erythrocyte binding to at least one of two DBP variants tested. Most (13 of 16) of these subjects still had inhibitory antibodies when reevaluated 12 months later. Cumulative exposure to malaria was the strongest predictor of DBP seropositivity identified by Multiple logistic regression models in this population. The poor antibody recognition of DBP elicited by natural exposure to P. vivax in Amazonian populations represents a challenge to be addressed by vaccine development strategies.

Limited global diversity of the Plasmodium vivax merozoite surface protein 4 gene

Merozoite surface proteins (MSPs) of the malaria parasites are major candidates for vaccine development targeting asexual blood stages. However, the diverse antigenic repertoire of these antigens that induce strain-specific protective immunity in human is a major challenge for vaccine design and often determines the efficacy of a vaccine. Here we further assessed the genetic diversity of Plasmodium vivax MSP4 (PvMSP4) protein using 195 parasite samples collected mostly from Thailand, Indonesia and Brazil. Overall, PvMSP4 is highly conserved with only eight amino acid substitutions. The majority of the haplotype diversity was restricted to the two short tetrapeptide repeat arrays in exon 1 and 2, respectively. Selection and neutrality tests indicated that exon 1 and the entire coding region of PvMSP4 were under purifying selection. Despite the limited nucleotide polymorphism of PvMSP4, significant genetic differentiation among the three major parasite populations was detected. Moreover, microgeographical heterogeneity was also evident in the parasite populations from different endemic areas of Thailand. (C) 2009 Elsevier B.V. All rights reserved.

The magnetized steel and scintillator calorimeters of the MINOS experiment

The Main Injector Neutrino Oscillation Search (MINOS) experiment uses an accelerator-produced neutrino beam to perform precision measurements of the neutrino oscillation parameters in the ""atmospheric neutrino"" sector associated with muon neutrino disappearance. This long-baseline experiment measures neutrino interactions in Fermilab`s NuMI neutrino beam with a near detector at Fermilab and again 735 km downstream with a far detector in the Soudan Underground Laboratory in northern Minnesota. The two detectors are magnetized steel-scintillator tracking calorimeters. They are designed to be as similar as possible in order to ensure that differences in detector response have minimal impact on the comparisons of event rates, energy spectra and topologies that are essential to MINOS measurements of oscillation parameters. The design, construction, calibration and performance of the far and near detectors are described in this paper. (C) 2008 Elsevier B.V. All rights reserved.

Thermoelectric transport properties through a T-shaped single quantum dot

We study the thermopower, thermal conductance, electric conductance and the thermoelectric figure of merit for a gate-defined T-shaped single quantum dot (QD). The QD is solved in the limit of strong Coulombian repulsion U -> infinity, inside the dot, and the quantum wire is modeled on a tight-binding linear chain. We employ the X-boson approach for the Anderson impurity model to describe the localized level within the quantum dot. Our results are in qualitative agreement with recent experimental reports and other theoretical researches for the case of a quantum dot embedded into a conduction channel, employing analogies between the two systems. The results for the thermopower sign as a function of the gate voltage (associated with the quantum dot energy) are in agreement with a recent experimental result obtained for a suspended quantum dot. The thermoelectric figure of merit times temperature results indicates that, at low temperatures and in the crossover between the intermediate valence and Kondo regimes, the system might have practical applicability in the development of thermoelectric devices. (c) 2010 Elsevier B.V. All rights reserved.

A review of the tectonic evolution of the Sunsas belt, SW Amazonian Craton

The Sunsas-Aguapei province (1.20-0.95 Ga), SW Amazonian Craton, is a key area to study the heterogeneous effects of collisional events with Laurentia, which shows evidence of the Grenvillian and Sunsas orogens. The Sunsas orogen, characterized by an allochthonous collisional-type belt (1.11-1.00 Ga), is the youngest and southwestern most of the events recorded along the cratonic fringe. Its evolution occurred after a period of long quiescence and erosion of the already cratonized provinces (>1.30 Ga), that led to sedimentation of the Sunsas and Vibosi groups in a passive margin setting. The passive margin stage was roughly contemporary with intraplate tectonics that produced the Nova Brasilandia proto-oceanic basin (<1.21 Ga), the reactivation of the Ji-Parana shear zone network (1.18-1.12 Ga) and a system of aborted rifts that evolved to the Huanchaca-Aguapei basin (1.17-1.15 Ga). The Sunsas belt is comprised by the metamorphosed Sunsas and Vibosi sequences, the Rincon del Tigre mafic-ultramafic sill and granitic intrusive suites. The latter rocks yield epsilon(Nd(t)) signatures (-0.5 to -4.5) and geochemistry (S,1, A-types) suggesting their origin associated with a continental arc setting. The Sunsas belt evolution is marked by ""tectonic fronts"" with sinistral offsets that was active from c. 1.08 to 1.05 Ga, along the southern edge of the Paragua microcontinent where K/Ar ages (1.27-1.34 Ga) and the Huanchaca-Aguapei flat-lying cover attest to the earliest tectonic stability at the time of the orogen. The Sunsas dynamics is coeval with inboard crustal shortening, transpression and magmatism in the Nova Brasilandia belt (1.13-1.00 Ga). Conversely, the Aguapei aulacogen (0.96-0.91 Ga) and nearby shear zones (0.93-0.91 Ga) are the late tectonic offshoots over the cratonic margin. The post-tectonic to anorogenic stages took place after ca. 1.00 Ga, evidenced by the occurrences of intra-plate A-type granites, pegmatites, mafic dikes and sills, as well as of graben basins. Integrated interpretation of the available data related to the Sunsas orogen supports the idea that the main nucleus of Rodinia incorporated the terrains forming the SW corner of Amazonia and most of the Grenvillian margin, as a result of two independent collisional events, as indicated in the Amazon region by the Ji-Parana shear zone event and the Sunsas belt, respectively. (C) 2009 Elsevier Ltd. All rights reserved.

THE RIO APA CRATON IN MATO GROSSO DO SUL (BRAZIL) AND NORTHERN PARAGUAY: GEOCHRONOLOGICAL EVOLUTION, CORRELATIONS AND TECTONIC IMPLICATIONS FOR RODINIA AND GONDWANA

The Rio Apa cratonic fragment crops out in Mato Grosso do Sul State of Brazil and in northeastern Paraguay. It comprises Paleo-Mesoproterozoic medium grade metamorphic rocks, intruded by granitic rocks, and is covered by the Neoproterozoic deposits of the Corumbi and Itapocurni Groups. Eastward it is bound by the southern portion of the Paraguay belt. In this work, more than 100 isotopic determinations, including U-Pb SHRIMP zircon ages, Rb-Sr and Sm-Nd whole-rock determinations, as well as K-Ar and Ar-Ar mineral ages, were reassessed in order to obtain a complete picture of its regional geological history. The tectonic evolution of the Rio Apa Craton starts with the formation of a series of magmatic arc complexes. The oldest U-Pb SHRIMP zircon age comes from a banded gneiss collected in the northern part of the region, with an age of 1950 +/- 23 Ma. The large granitic intrusion of the Alumiador Batholith yielded a U-Pb zircon age of 1839 +/- 33 Ma, and from the southeastern part of the area two orthogneisses gave zircon U-Pb ages of 1774 +/- 26 Ma and 1721 +/- 25 Ma. These may be coeval with the Alto Terere metamorphic rocks of the northeastern corner, intruded in their turn by the Baia das Garcas granitic rocks, one of them yielding a zircon U-Pb age of 1754 +/- 49 Ma. The original magmatic protoliths of these rocks involved some crustal component, as indicated by the Sm-Nd TDm model ages, between 1.9 and 2.5 Ga. Regional Sr isotopic homogenization, associated with tectonic deformation and medium-grade metamorphism occurred at approximately 1670 Ma, as suggested by Rb-Sr whole rock reference isochrons. Finally, at 1300 Ma ago, the Ar work indicates that the Rio Apa Craton was affected by widespread regional heating, when the temperature probably exceeded 350 degrees C. Geographic distribution, age and isotopic signature of the fithotectonic units suggest the existence of a major suture separating two different tectonic domains, juxtaposed at about 1670 Ma. From that time on, the unified Rio Apa continental block behaved as one coherent and stable tectonic unit. It correlates well with the SW corner of the Amazonian Craton, where the medium-grade rocks of the Juruena-Rio Negro tectonic province, with ages between 1600 and 1780 Ma, were reworked at about 1300 Ma. Looking at the largest scale, the Rio Apa Craton is probably attached to the larger Amazonian Craton, and the actual configuration of southwestern South America is possibly due to a complex arrangement of allochthonous blocks such as the Arequipa, Antofalla and Pampia, with different sizes, that may have originated as disrupted parts of either Laurentia or Amazonia, and were trapped during later collisions of these continental masses.

The position of the Amazonian Craton in supercontinents

This paper examines the extensive regions of Proterozoic accretionary belts that either formed most of the Amazonian Craton, or are marginal to its southeastern border. Their overall geodynamic significance is considered taking into account the paleogeographic reconstruction of Columbia, Rodinia and Gondwana. Amazonia would be part of Columbia together With Laurentia, North China and Baltica, forming a continuous, continental landmass linked by the Paleo- to Mesoproterozoic mobile belts that constitute large portions of it. The Rodinia supercontinent was formed in the Mesoproterozoic by the agglutination of the existing cratonic fragments, such as Laurentia and Amazonia, during contemporary continental collisions worldwide. The available paleomagnetic data suggest that Laurentia and Amazonia remained attached until at least 600 Ma. Since all other cratonic units Surrounding Laurentia have already rifted away by that time, the separation between Amazonia and Laurentia marks the final break-up of Rodinia with the opening of the lapetus ocean. (C) 2009 International Association for Gondwana Research. Published by Elsevier B.V. All rights reserved.

Equicrestal and Subcrestal Dental Implants: A Histologic and Histomorphometric Evaluation of Nine Retrieved Human Implants

Background: Stability of pen-implant crestal bone plays a relevant role relative to the presence or absence of interdental papilla. Several factors can contribute to the crestal bone resorption observed around two-piece implants, such as the presence of a microgap at the level of the implant abutment junction, the type of connection between implant and prosthetic components, the implant positioning relative to the alveolar crest, and the interimplant distance. Subcrestal positioning of dental implants has been proposed to decrease the risk of exposure of the metal of the top of the implant or of the abutment margin, and to get enough space in a vertical dimension to create a harmoniously esthetic emergence profile. Methods: The present retrospective histologic study was performed to evaluate dental implants retrieved from human jaws that had been inserted in an equicrestal or subcrestal position. A total of nine implants were evaluated: five of these had been inserted in an equicrestal position, whereas the other four had been positioned subcrestally (1 to 3 mm). Results: In all subcrestally placed implants, preexisting and newly formed bone was found over the implant shoulder. In the equicrestal implants, crestal bone resorption (0.5 to 1.5 mm) was present around all implants. Conclusion: The subcrestal position of the implants resulted in bone located above the implant shoulder. J Periodontol 2011;82:708-715.

Osteocyte Density in the Peri-Implant Bone of Immediately Loaded and Submerged Dental Implants

Background: The role of osteocytes in bone structure and function remains partially unresolved. Their participation in mechanotransduction, i.e., the conversion of a physical stimulus into a cellular response, has been hypothesized. The present study was an evaluation of the osteocyte density in the peri-implant bone of immediately loaded and submerged dental implants. Methods: Fourteen male patients were included in the study; all of them were partially edentulous and needed a posterior mandibular restoration. Implants were inserted in these areas; half of the sample was loaded immediately (included in a fixed provisional prosthesis on the same day as implant surgery), whereas the other half was left to heal submerged. Fourteen implants (seven immediately loaded and seven unloaded) were retrieved with a trephine after a healing period of 8 weeks. The specimens were treated to obtain thin ground sections, and histomorphometry was used to evaluate the osteocyte index in the peri-implant bone. Results: A higher and statistically significant number of osteocytes was found in the peri-implant bone around immediately loaded implants (P=0.0081). A correlation between the percentage of bone-implant contact and osteocyte density was found for immediately loaded implants (P=0.0480) but not for submerged implants (P=0.2667). Conclusion: The higher number of osteocytes in the peri-implant bone around immediately loaded implants could be related to the functional adaptation required by the loading stimulus, which also explains the hypothesized involvement of the osteocytes in the maintenance of the bone matrix. J Periodontol 2009;80:499-504.

Health in Brazil 5 Violence and injuries in Brazil: the effect, progress made, and challenges ahead

Although there are signs of decline, homicides and traffic-related injuries and deaths in Brazil account for almost two-thirds of all deaths from external causes. In 2007, the homicide rate was 26.8 per 100 000 people and traffic-related mortality was 23.5 per 100 000. Domestic violence might not lead to as many deaths, but its share of violence-related morbidity is large. These are important public health problems that lead to enormous individual and collective costs. Young, black, and poor men are the main victims and perpetrators of community violence, whereas poor black women and children are the main victims of domestic violence. Regional differentials are also substantial. Besides the sociocultural determinants, much of the violence in Brazil has been associated with the misuse of alcohol and illicit drugs, and the wide availability of firearms. The high traffic-related morbidity and mortality in Brazil have been linked to the chosen model for the transport system that has given priority to roads and private-car use without offering adequate infrastructure. The system is often poorly equipped to deal with violations of traffic rules. In response to the major problems of violence and injuries, Brazil has greatly advanced in terms of legislation and action plans. The main challenge is to assess these advances to identify, extend, integrate, and continue the successful ones.

Socio-economic variables influence the prevalence of inadequate nutrient intake in Brazilian adolescents: results from a population-based survey

Objective: To estimate the prevalence of inadequate nutrient intake among adolescents and the association between socio-economic variables and nutritional status. Design: Cross-sectional study with a population-based sample. Settings: The usual nutrient intake distribution was estimated using the Iowa State University method. The Estimated Average Requirement cut-off point method was used to determine the proportion of adolescents with inadequate intake for each nutrient, according to sex, income, parental educational level and nutritional status. Subjects: Twenty-four-hour dietary recalls were applied in 525 male and female Brazilian adolescents aged 14-18 years. Results: The highest prevalence of inadequate nutrient intake was observed for vitamin E (99% in both sexes). For male and female adolescents, the prevalence of inadequate intake was: Mg, 89% and 84%; vitamin A, 78% and 71 %; vitamin C, 79% and 53%; and vitamin B(6), 21% and 33%, respectively. The prevalence of inadequate intake for niacin, thiamin, riboflavin, Se, Cu and vitamin B(12) was <15 %. Individuals in the lower income and lower parental educational level strata had the highest risk of having inadequate intake for P, riboflavin and vitamins A, B(6) and B(12). Compared with non-overweight individuals, overweight individuals had a higher risk of inadequate intake for Mg, vitamin A, P, thiamin and riboflavin. Conclusions: The present study found a high prevalence of inadequate intake of nutrients that are recognised as being protective against chronic diseases. Adolescents in the lower income and lower parental educational level strata were less likely to have their nutrient intake requirements met.

New perspectives on the synoptic and mesoscale structure of Hurricane Catarina

This work explores in detail synoptic and mesoscale features of Hurricane Catarina during its life cycle from a decaying baroclinic wave to a tropical depression that underwent tropical transition (TT) and finally to a Category 2 hurricane at landfall over Santa Catarina State coast, southern Brazil. This unique system caused 11 deaths mostly off the Brazilian coast and an estimated half billion dollars in damage in a matter of a few hours on 28 March 2004. Although the closest meteorological station available was tens of kilometres away from the eye, in situ meteorological measurements provided by a work-team sent to the area where the eye made landfall unequivocally reproduces the tropical signature with category 2 strength, adding to previous analysis where this data was not available. Further analyses are based mostly on remote sensing data available at the time of the event. A classic dipole blocking set synoptic conditions for Hurricane Catarina to develop, dynamically contributing to the low wind shear observed. On the other hand, on its westward transit, large scale subsidence limited its strength and vertical development. Catarina had relatively cool SST conditions, but this was mitigated by favourable air-sea fluxes leading to latent heat release-driven processes during the mature phase. The ocean`s dynamic topography also suggested the presence of nearby warm core rings which may have facilitated the transition and post-transition intensification. Since there were no records of such a system at least in the past 30 years and given that SSTs were generally below 26 degrees C and vertical shear was usually strong, despite all satellite data available, the system was initially classified as an extratropical cyclone. Here we hypothesise that this categorization was based oil inadequate regional scale model outputs which did not account for the importance of the latent heat fluxes over the ocean. Hurricane Catarina represents a dramatic event on weather systems in South America. It has attracted attention worldwide and poses questions as whether or not it is a symptom of global warming. (C) 2009 Elsevier B.V. All rights reserved.