Coronavirus mRNA synthesis: identification of novel transcription initiation signals which are differentially regulated by different leader sequences.

The mRNA synthesis of mouse hepatitis virus (MHV) has been proposed to be the result of interaction between the leader RNA and the intergenic sites. Previously, we have identified a transcription initiation site (for mRNA 2-1), which is more efficiently transcribed by viruses containing two copies of UCUAA sequence in the leader RNA than by those with three copies. In this study, we have identified several sites which are regulated in the opposite way, namely, they are efficiently transcribed by the leader RNA with three UCUAA copies but not by those with two copies. These sites were characterized by primer extension and amplification by polymerase chain reaction. One of these sites is in the gene 3 region of a recombinant virus between A59 and JHM strains of MHV. Another is in the gene 2 region of MHV-1 strain. Both of these sites have a sequence similar to but different from the consensus transcription initiation signal (UCUAAUCUAUC and UUUAAUCUU, as opposed to UCUAAAC). These two novel intergenic sequences are not present in the genome of the JHM strain, consistent with the absence of these mRNAs in the JHM-infected cells. The discovery of this type of transcription initiation site provides additional evidence for the importance of the leader RNA in the transcription initiation of MHV mRNAs

High bioaccumulation of cadmium and other metals in Patagonian edible gastropods

High concentrations of metals are accumulated by edible marine resources, affecting human health. Marine gastropods have been commercially captured and consumed in Argentina for decades without official regulations or studies to guarantee their safety. In this study, metals (cadmium, aluminium, iron, zinc, copper and lead) were analysed for the first time in the edible marine gastropods Buccinanops globulosus, Adelomelon ancilla and Trophon geversianus and their surrounding sediments. Taking into account the maximum levels recommended by national and international regulations for food safety, we recommend consuming only the foot of these gastropod species. We recommend not consuming these gastropods from harbour areas due to concentrations of lead in sediments and tissues. Though the target hazard quotient (THQ) of cadmium was not high for the local community, it could be higher for other populations who consume molluscs more frequently, indicating a potential health risk of chronic exposure. Monitoring of these and other contaminants should be performed in order to ensure food safety of these marine resources.Fil: Primost, Monica Angelina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto de Biología de Organismos Marinos; ArgentinaFil: Gil, Mónica N.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Centro para el Estudio de Sistemas Marinos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico; ArgentinaFil: Bigatti, Gregorio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto de Biología de Organismos Marinos; Argentina. Universidad Nacional de la Patagonia; Argentin

Surface Plasmon Resonance kinetic analysis of the interaction between G-quadruplex nucleic acids and an anti-G-quadruplex monoclonal antibody

Background G-quadruplexes (G4s) are nucleic acids secondary structures formed in guanine-rich sequences. Anti-G4 antibodies represent a tool for the direct investigation of G4s in cells. Surface Plasmon Resonance (SPR) is a highly sensitive technology, suitable for assessing the affinity between biomolecules. We here aimed at improving the orientation of an anti-G4 antibody on the SPR sensor chip to optimize detection of binding antigens. Methods SPR was employed to characterize the anti-G4 antibody interaction with G4 and non-G4 oligonucleotides. Dextran-functionalized sensor chips were used both in covalent coupling and capturing procedures. Results The use of two leading molecule for orienting the antibody of interest allowed to improve its activity from completely non-functional to 65% active. The specificity of the anti-G4 antobody for G4 structures could thus be assessed with high sensitivity and reliability. Conclusions Optimization of the immobilization protocol for SPR biosensing, allowed us to determine the anti-G4 antibody affinity and specificity for G4 antigens with higher sensitivity with respect to other in vitro assays such as ELISA. Anti-G4 antibody specificity is a fundamental assumption for the future utilization of this kind of antibodies for monitoring G4s directly in cells. General significance The heterogeneous orientation of amine-coupling immobilized ligands is a general problem that often leads to partial or complete inactivation of the molecules. Here we describe a new strategy for improving ligand orientation: driving it from two sides. This principle can be virtually applied to every molecule that loses its activity or is poorly immobilized after standard coupling to the SPR chip surface

Biosynthesis, structure, and biological activities of envelope protein gp65 of murine coronavirus.

We have previously shown that gp65 (E3) is a virion structural protein which varies widely in quantity among different strains of mouse hepatitis virus (MHV). In this study, the biosynthetic pathway and possible biological activities of this protein were examined. The glycosylation of gp65 in virus-infected cells was inhibited by tunicamycin but not by monensin, suggesting that it contains an N-glycosidic linkage. Glycosylation is cotranslational and appears to be complete before the glycoprotein reaches the Golgi complex. Pulse-chase experiments showed that this protein decreased in size after 30 min of chase, suggesting that the carbohydrate chains of gp65 undergo trimming during its transport across the Golgi. This interpretation is supported by the endoglycosidase treatment of gp65, which showed that the peptide backbone of gp65 did not decrease in size after pulse-chase periods. This maturation pathway is distinct from that of the E1 or E2 glycoproteins. Partial endoglycosidase treatment indicated that gp65 contains 9 to 10 carbohydrate side chains; thus, almost all of the potential glycosylation sites of gp65 were glycosylated. In vitro translation studies coupled with protease digestion suggest that gp65 is an integral membrane protein. The presence of gp65 in the virion is correlated with the presence of an acetylesterase activity. No hemagglutinin activity was detected

Designing Leadership Development Program Curriculum: A Narrative Research Study

A high priority is placed on developing leadership skills because of its importance to organizational performance and growth. Since 2010, significant investment has been put into leadership programs by companies globally with the goal of producing more effective leaders. Despite the expenditure into leadership development, most training efforts fail to meet learners’ needs because they do not include participant perspectives in the content decision-making process, using outdated leadership theories as a framework for content development, or failing to evaluate training efforts. These failures result in a mismatch between the challenges leaders face in their roles and the content of a leadership program. Therefore, to successfully design programs that meet learners’ needs, an understanding of an audience’s challenges is critical. The purpose of this qualitative study was to understand the challenges leaders face in a global technology company. In this study, the researcher specifically used narrative inquiry to understand the stories of past participants in a leadership development program. The researcher collected data by retrospective analyses of open-ended internal surveys gathered by the study organization during the last 12 months for the purposes of identifying challenges leaders face in their leadership roles and understanding perceptions of how well a leadership program prepared them to respond to these challenges. The researcher intends the findings to help the organization identify to what extent the content of their current leadership program addressed the challenges of the participants

ENHANCING THE EFFECTIVENESS OF POVERTY REDUCTION INITIATIVE USING INFORMATION TECHNOLOGY

Poverty seems to have persisted in many developing countries like Nigeria despite various measures and strategiesthat have been adopted over the years to deal with it. In spite of these efforts majority of the populace is povertystricken. The problem is not introducing these initiatives but rather how effective these initiatives are in reducingpoverty. Many governments in Nigeria have introduced various strategies/initiatives to reduce poverty but thestrategies seem not to have been effective in dealing with the problem. There is need therefore to find a way ofenhancing the effectiveness of these initiatives. The paper is focused on how IT can be used to enhance theeffectiveness of the different poverty initiatives so as to reduce poverty to the barest minimum.Keywords: Information Technology, Poverty , Poverty Reduction Initiatives

Effect of Aerobic Activities on Promoting Healthy Fitness Standards of Freshman Students

Obesity is one of the greatest threats to child and adult health in the United States. Adolescents are engaging in sedentary behaviors, and their body fat composition is directly affected by a lack of exercise. Direct instruction was used as the framework to compare the results.https://scholarworks.waldenu.edu/archivedposters/1077/thumbnail.jp

A Cross-National Analysis of the Nutrition Habits of Hispanic Mothers and Daughters

The rates of obesity in Hispanic women increased significantly between 1994 and 2008 from 35.3% to 45.1% (National Center for Health Statistics, 2010). Poor nutritional habits and obesity have longterm negative health ramifications that warrant targeted efforts to stem this growing epidemic. Two articles are included in this portfolio. The first article discusses the state of the science in relation to cultural aspects of nutrition choices of Hispanic women. The aim of this article is to set the context for the cross-national study which is the focus of the second article. This study examines current eating habits, weight history, health perception, future time perspective, family dietary support, friend dietary support, and the health outcomes of BMI, waist circumference, and nutrition of 157 Hispanic daughters and mothers in Texas and Mexico. The differences between U.S. and Mexico cohorts are also examined in relation to their nutrition choices and their generational preferences. This work indicates that the younger generation appears to have nutrition habits more closely associated with their age cohort than their family unit. In the context of the surging epidemic of obesity in the Hispanic culture and with an awareness of the pivotal role played by the Hispanic woman in the health and nutrition choices of her family, this research project provides an initial dialogue regarding factors influencing the Hispanic population toward healthier eating habits and increased health promoting behaviors