The Immunomodulatory Action of Sialostatin L on Dendritic Cells Reveals Its Potential to Interfere with Autoimmunity

Sialostatin L (SialoL) is a secreted cysteine protease inhibitor identified in the salivary glands of the Lyme disease vector Ixodes scapularis. In this study, we reveal the mechanisms of SialoL immunomodulatory actions on the vertebrate host. LPS-induced maturation of dendritic cells from C57BL/6 mice was significantly reduced in the presence of SialoL. Although OVA degradation was not affected by the presence of SialoL in dendritic cell cultures, cathepsin S activity was partially inhibited, leading to an accumulation of a 10-kDa invariant chain intermediate in these cells. As a consequence, in vitro Ag-specific CD4(+) T cell proliferation was inhibited in a time-dependent manner by SialoL, and further studies engaging cathepsin S(-/-) or cathepsin L(-/-) dendritic cells confirmed that the immunomodulatory actions of SialoL are mediated by inhibition of cathepsin S. Moreover, mice treated with SialoL displayed decreased early T cell expansion and recall response upon antigenic stimulation. Finally, SialoL administration during the immunization phase of experimental autoimmune encephalomyelitis in mice significantly prevented disease symptoms, which was associated with impaired IFN-gamma and IL-17 production and specific T cell proliferation. These results illuminate the dual mechanism by which a human disease vector protein modulates vertebrate host immunity and reveals its potential in prevention of an autoimmune disease. The Journal of Immunology, 2009, 182: 7422-7429.

Interaction of Mycoplasma genitalium with host cells: evidence for nuclear localization

Mycoplasma genitalium (Mg) is a mollicute that causes a range of human urogenital infections. A hallmark of these bacteria is their ability to establish chronic infections that can persist despite completion of appropriate antibiotic therapies and intact and functional immune systems. Intimate adherence and surface colonization of mycoplasmas to host cells are important pathogenic features. However, their facultative intracellular nature is poorly understood, partly due to difficulties in developing and standardizing cellular interaction model systems. Here, we characterize growth and invasion properties of two Mg strains (G37 and 1019V). Mg G37 is a high-passage laboratory strain, while Mg 1019V is a low-passage isolate recovered from the cervix. The two strains diverge partially in gene sequences for adherence-related proteins and exhibit subtle variations in their axenic growth. However, with both strains and consistent with our previous studies, a subset of adherent Mg organisms invade host cells and exhibit perinuclear targeting. Remarkably, intranuclear localization of Mg proteins is observed, which occurred as early as 30 min after infection. Mg strains deficient in adherence were markedly reduced in their ability to invade and associate with perinuclear and nuclear sites.

Population genetic analysis of large sequence polymorphisms in Plasmodium falciparum blood-stage antigens

Plasmodium falciparum, the causative agent of human malaria, invades host erythrocytes using several proteins on the surface of the invasive merozoite, which have been proposed as potential vaccine candidates. Members of the multi-gene PfRh family are surface antigens that have been shown to play a central role in directing merozoites to alternative erythrocyte receptors for invasion. Recently, we identified a large structural polymorphism, a 0.58 Kb deletion, in the C-terminal region of the PfRh2b gene, present at a high frequency in parasite populations from Senegal. We hypothesize that this region is a target of humoral immunity. Here, by analyzing 371 P. falciparum isolates we show that this major allele is present at varying frequencies in different populations within Senegal, Africa, and throughout the world. For allelic dimorphisms in the asexual stage antigens, Msp-2 and EBA-175, we find minimal geographic differentiation among parasite populations from Senegal and other African localities, suggesting extensive gene flow among these populations and/or immune-mediated frequency-dependent balancing selection. In contrast, we observe a higher level of inter-population divergence (as measured by F(st)) for the PfRh2b deletion, similar to that observed for SNPs from the sexual stage Pfs45/48 loci, which is postulated to be under directional selection. We confirm that the region containing the PfRh2b polymorphism is a target of humoral immune responses by demonstrating antibody reactivity of endemic sera. Our analysis of inter-population divergence suggests that in contrast to the large allelic dimorphisms in EBA-175 and Msp-2, the presence or absence of the large PfRh2b deletion may not elicit frequency-dependent immune selection, but may be under positive immune selection, having important implications for the development of these proteins as vaccine candidates. (C) 2009 Elsevier B.V. All rights reserved.

Comparative genomics of the neglected human malaria parasite Plasmodium vivax

The human malaria parasite Plasmodium vivax is responsible for 25 - 40% of the similar to 515 million annual cases of malaria worldwide. Although seldom fatal, the parasite elicits severe and incapacitating clinical symptoms and often causes relapses months after a primary infection has cleared. Despite its importance as a major human pathogen, P. vivax is little studied because it cannot be propagated continuously in the laboratory except in non- human primates. We sequenced the genome of P. vivax to shed light on its distinctive biological features, and as a means to drive development of new drugs and vaccines. Here we describe the synteny and isochore structure of P. vivax chromosomes, and show that the parasite resembles other malaria parasites in gene content and metabolic potential, but possesses novel gene families and potential alternative invasion pathways not recognized previously. Completion of the P. vivax genome provides the scientific community with a valuable resource that can be used to advance investigation into this neglected species.

Endurance training activates AMP-activated protein kinase, increases expression of uncoupling protein 2 and reduces insulin secretion from rat pancreatic islets

Endurance exercise is known to enhance peripheral insulin sensitivity and reduce insulin secretion. However, it is unknown whether the latter effect is due to the reduction in plasma substrate availability or alterations in beta-cell secretory machinery. Here, we tested the hypothesis that endurance exercise reduces insulin secretion by altering the intracellular energy-sensitive AMP-activated kinase (AMPK) signaling pathway. Male Wistar rats were submitted to endurance protocol training one, three, or five times per week, over 8 weeks. After that, pancreatic islets were isolated, and glucose-induced insulin secretion (GIIS), glucose transporter 2 (GLUT2) protein content, total and phosphorylated calmodulin kinase kinase (CaMKII), and AMPK levels as well as peroxisome proliferator-activated receptor-gamma coactivator-1-alpha (PGC-1 alpha) and uncoupling protein 2 (UCP2) content were measured. After 8 weeks, chronic endurance exercise reduced GIIS in a dose-response manner proportionally to weekly exercise frequency. Contrariwise, increases in GLUT2 protein content, CaMKII and AMPK phosphorylation levels were observed. These alterations were accompanied by an increase in UCP2 content, probably mediated by an enhancement in PGC-1 alpha protein expression. In conclusion, chronic endurance exercise induces adaptations in beta-cells leading to a reduction in GIIS, probably by activating the AMPK signaling pathway. Journal of Endocrinology (2011) 208, 257-264

Analysis of Agonist and Antagonist Effects on Thyroid Hormone Receptor Conformation by Hydrogen/Deuterium Exchange

Thyroid hormone receptors (TRs) are ligand-gated transcription factors with critical roles in development and metabolism. Although x-ray structures of TR ligand-binding domains (LBDs) with agonists are available, comparable structures without ligand (apo-TR) or with antagonists are not. It remains important to understand apo-LBD conformation and the way that it rearranges with ligands to develop better TR pharmaceuticals. In this study, we conducted hydrogen/deuterium exchange on TR LBDs with or without agonist (T(3)) or antagonist (NH(3)). Both ligands reduce deuterium incorporation into LBD amide hydrogens, implying tighter overall folding of the domain. As predicted, mass spectroscopic analysis of individual proteolytic peptides after hydrogen/deuterium exchange reveals that ligand increases the degree of solvent protection of regions close to the buried ligand-binding pocket. However, there is also extensive ligand protection of other regions, including the dimer surface at H10-H11, providing evidence for allosteric communication between the ligand-binding pocket and distant interaction surfaces. Surprisingly, C-terminal activation helix H12, which is known to alter position with ligand, remains relatively protected from solvent in all conditions suggesting that it is packed against the LBD irrespective of the presence or type of ligand. T(3), but not NH(3), increases accessibility of the upper part of H3-H5 to solvent, and we propose that TR H12 interacts with this region in apo-TR and that this interaction is blocked by T(3) but not NH(3.) We present data from site-directed mutagenesis experiments and molecular dynamics simulations that lend support to this structural model of apo-TR and its ligand-dependent conformational changes. (Molecular Endocrinology 25: 15-31, 2011)

Actions of octocoral and tobacco cembranoids on nicotinic receptors

Nicotinic acetylcholine receptors (AChRs) are pentameric proteins that form agonist-gated cation channels through the plasma membrane. AChR agonists and antagonists are potential candidates for the treatment of neurodegenerative diseases. Cembranoids are naturally occurring diterpenoids that contain a 14-carbon ring. These diterpenoids interact with AChRs in complex ways: as irreversible inhibitors at the agonist sites, as noncompetitive inhibitors, or as positive modulators, but no cembranoid was ever shown to have agonistic activity on AChRs. The cembranoid eupalmerin acetate displays positive modulation of agonist-induced currents in the muscle-type AChR and in the related gamma-aminobutyric acid (GABA) type A receptor. Moreover, cembranoids display important biological effects, many of them mediated by nicotinic receptors. Cembranoids from tobacco are neuroprotective through a nicotinic anti-apoptotic mechanism preventing excitotoxic neuronal death which in part could result from anti-inflammatory properties of cembranoids. Moreover, tobacco cembranoids also have anti-inflammatory properties which could enhance their neuroprotective properties. Cembranoids from tobacco affect nicotine-related behavior: they increase the transient initial ataxia caused by first nicotine injection into naive rats and inhibit the expression of locomotor sensitization to repeated injections of nicotine. In addition, cembranoids are known to act as anti-tumor compounds. In conclusion, cembranoids provide a promising source of lead drugs for many clinical areas, including neuroprotection, smoking-cessation, and anti-cancer therapies. (C) 2009 Elsevier Ltd. All rights reserved.

Self-reported conflict management competence among nursing students on the point of graduating and registered nurses with professional experience

Objective: It has been shown that specific competence is necessary for preventing and managing conflicts in healthcare settings. The aim of this descriptive and correlation study was to investigate and compare the self-reported conflict management competence (CMC) of nursing students who were on the point of graduating (NSPGs), and the CMC of registered nurses (RNs) with professional experience. Methods: The data collection, which consisted of soliciting answers to items measuring CMC in the Nurse Professional Competence (NPC) Scale, was performed as a purposive selection of 11 higher education institutions (HEIs) in Sweden. Three CMC items from the NPC Scale were answered by a total of 569 nursing students who were on the point of graduating and 227 RN registered nurses with professional experience. Results: No significant differences between NSPGs and RNs were found, and both groups showed a similar score pattern, with the lowest score for the item: “How do you perceive your ability to develop the group and strengthen competence in conflict management and problem-solving, based on knowledge of group dynamics?”. RNs with long professional experience (>24 months) rated their overall CMC as significantly better than RNs with short (<24 months) professional experience did (p = .05). NSPGs who had experience of international studies during their nursing education reported higher CMC, compared with those who did not have this experience (p = .03). RNs who reported a high degree of utilisation of CMC during the previous month scored higher regarding self-reported overall CMC (p < .0001). Conclusions: Experience of international studies during nursing education, or long professional experience, resulted in higher self-reported CMC. Hence, the CMC items in the NPC Scale can be suitable for identifying self-reported conflict management competence among NSPGs and RNs

Aquatic macrophytes of Itaipu Reservoir, Brazil: Survey of species and ecological considerations

In a survey of the aquatic macrophytes of the Itaipu Reservoir, we identified 62 taxa in 25 families and 42 genera. The highest number taxa was observed for the emergent macrophytes (40 taxa). Reduced fluctuation in water level, increased light penetration, and sediment enrichment by nutrients and organic matter following the formation of the reservoir favored the appearance of a species-rich community of submerged macrophytes (23% of the taxa identified). The aquatic macrophytes were found mainly near the mouths of the main tributaries of the reservoir, in shallow area of depth less than 2 meters. In addition to the shallow depth, the greater nutrient input from the tributaries and relative protection from wind explained this distribution. Among the species found, Egeria najas merits mention for its occurrence in all localities sampled, with biomass values varying between 98 and 186 gDW/m2. Some potential nuisance species such as Eichhornia crassipes, Salvinia auriculata, and Pistia stratiotes also deserve attention, since they were also observed to be covering large areas of Itaipu Reservoir. © 1999, Paraná Institute of Technology Publication.

A new generation of standards: implications for the Caribbean and Latin America

Incluye Bibliografía

Modernization and new services of the Brazilian active control network

The Brazilian Network for Continuous Monitoring of GNSS - RBMC is a national network of continuously operating reference GNSS stations. Since its establishment in December of 1996, it has been playing an essential role for the maintenance and user access of the fundamental geodetic frame in the country. In order to provide better services for RBMC, the Brazilian Institute of Geography and Statistics - IBGE and the National Institute of Colonization and Land Reform - INCRA are both partners involved in the National Geospatial Framework Project - PIGN. This paper provides an overview of the recent modernization phases the RBMC network has undergone highlighting its future steps. These steps involve the installation of new equipment, provide real time data from a group of core stations and compute real-time DGPS corrections, based on CDGPS (The real-time Canada-Wide DGPS Service) (The Real-Time Canada-Wide DGPS Service. http://www.cdgps.com/ 2009a). In addition to this, a post-mission Precise Point Positioning (PPP) service has been established based on the current Geodetic Survey Division of NRCan (CSRS-PPP) service. This service is operational since April 2009 and is in large use in the country. All activities mentioned before are based on a cooperation signed at the end of 2004 with the University of New Brunswick, supported by the Canadian International Development Agency and the Brazilian Cooperation Agency. The Geodetic Survey Division of NRCan is also participating in this modernization effort under the same project. This infrastructure of 66 GNSS stations, the real time, post processing services and the potentiality of providing Wide Area DGPS corrections in the future show that the RBMC system is comparable to those available in USA and Europe. © Springer-Verlag Berlin Heidelberg 2012.

Comparative study of policies on technology and industry in the Caribbean and their effects on development

Includes bibliography

Causes and consequences of low rates of specialisation in science and technology in CDCC member countries

The importance of science and technology (S&T) in Small Island Developing States (SIDS) is clearly articulated in Chapter XI, paragraphs 57, 58, 61 and 62 of the Mauritius Strategy for the Further Implementation of the Programme of Action for Sustainable Development of Small Island Developing States (MSI). At the regional level, the Heads of Government of the Caribbean Community (CARICOM) noted the challenge that CARICOM member States face in competing in this new international economic environment in which the impact of scientific and technological change has created a knowledge-based global economy. Given the importance of S&T to development of Caribbean SIDS, the Economic Commission for Latin America and the Caribbean (ECLAC) Subregional Headquarters for the Caribbean embarked on a study to determine the causes and consequences of low rates of specialisation in S&T with a view to making recommendations for development of strategies for addressing these challenges. Data on postgraduate (Master of Science, Master of Philosophy and Doctor of Philosophy) enrolment and graduation in agriculture, engineering and the sciences from the three campuses of the University of the West Indies (UWI) as well as from the University of Technology in Jamaica and the University of Trinidad and Tobago (UTT) were examined and analysed. Face-to-face interviews were also held with key personnel from these institutions and a questionnaire was also served to individuals in key institutions. Results of the study revealed that although the number of students enrolled in higher degree programmes has increased in absolute terms, they are decreasing in relative terms. However, enrolment in agriculture has indeed declined while enrolment rates in engineering, although increasing, were not significantly high. Market forces have proved to be a main reason for this trend while facilities for the conduct and supervision of cutting-edge research, the disconnect between science and industry and societal labelling of scientists as “misfits” are also contributing to the situation. This has resulted in a reduced desire by students at all levels of the school system and faculty to be involved in S&T; lack of innovation; a better staffed private, as compared with public, sector; and poor remuneration in science-based employment. There also appears to be a gender bias in enrolment with more males than females being enrolled in engineering while the opposite is apparent in agriculture and the sciences. Recommendations for remedying this situation range from increasing investment in S&T, creating linkages between science and industry as well as with the international community, raising awareness of the value of S&T at all levels of the education system to informing policy to stimulate the science – innovation interface so as to promote intellectual property rights.

Lufaxin, a Novel Factor Xa Inhibitor From the Salivary Gland of the Sand Fly Lutzomyia longipalpis Blocks Protease-Activated Receptor 2 Activation and Inhibits Inflammation and Thrombosis In Vivo

Objective-Blood-sucking arthropods' salivary glands contain a remarkable diversity of antihemostatics. The aim of the present study was to identify the unique salivary anticoagulant of the sand fly Lutzomyia longipalpis, which remained elusive for decades. Methods and Results-Several L. longipalpis salivary proteins were expressed in human embryonic kidney 293 cells and screened for inhibition of blood coagulation. A novel 32.4-kDa molecule, named Lufaxin, was identified as a slow, tight, noncompetitive, and reversible inhibitor of factor Xa (FXa). Notably, Lufaxin's primary sequence does not share similarity to any physiological or salivary inhibitors of coagulation reported to date. Lufaxin is specific for FXa and does not interact with FX, Dansyl-Glu-Gly-Arg-FXa, or 15 other enzymes. In addition, Lufaxin blocks prothrombinase and increases both prothrombin time and activated partial thromboplastin time. Surface plasmon resonance experiments revealed that FXa binds Lufaxin with an equilibrium constant approximate to 3 nM, and isothermal titration calorimetry determined a stoichiometry of 1:1. Lufaxin also prevents protease-activated receptor 2 activation by FXa in the MDA-MB-231 cell line and abrogates edema formation triggered by injection of FXa in the paw of mice. Moreover, Lufaxin prevents FeCl3-induced carotid artery thrombus formation and prolongs activated partial thromboplastin time ex vivo, implying that it works as an anticoagulant in vivo. Finally, salivary gland of sand flies was found to inhibit FXa and to interact with the enzyme. Conclusion-Lufaxin belongs to a novel family of slow-tight FXa inhibitors, which display antithrombotic and anti-inflammatory activities. It is a useful tool to understand FXa structural features and its role in prohemostatic and proinflammatory events. (Arterioscler Thromb Vasc Biol. 2012;32:2185-2196.)

Influence of HLA-DRB-1 alleles on the production of antibody against CSP, MSP-1, AMA-1, and DBP in Brazilian individuals naturally infected with Plasmodium vivax

We evaluated the influence of allelic frequency of the human leukocyte antigen (HLA) -DRB1 on the acquisition of antibody response against malaria sporozoite and merozoite peptides in patients with Plasmodium vivax malaria acquired in endemic areas of Brazil. IgG antibodies were detected by enzyme-linked immunosorbent assay against four peptides of circumsporozoite protein (CSP) (amino, carboxyl, and VK210 and VK247 repeats) and peptides of merozoite surface protein 1 (MSP-1), apical membrane antigen 1 (AMA-1), and Duffy-binding protein (DBP). We found an association between HLA-DR3 and HLA-DR5 alleles and lack of antibody response to CSP amino terminal, as well as an association between HILA-DR3 and the highest antibody response to MSP1 (Pv200L). In conclusion, we suggest a potential regulatory role of the H1A-DRB1 alleles in the production of antibodies to a conserved region of P. vivax CSP and MSP1 in Brazilian population exposed to malaria. (C) 2011 Elsevier B.V. All rights reserved.