Use of first nucleotide change technology to determine the frequency of factor V Leiden in a population of Australian blood donors

Activated protein C resistance (APCR), the most common risk factor for venous thrombosis, is the result of a G to A base substitution at nucleotide 1691 (R506Q) in the factor V gene. Current techniques to detect the factor V Leiden mutation, such as determination of restriction length polymorphisms, do not have the capacity to screen large numbers of samples in a rapid, cost- effective test. The aim of this study was to apply the first nucleotide change (FNC) technology, to the detection of the factor V Leiden mutation. After preliminary amplification of genomic DNA by polymerase chain reaction (PCR), an allele-specific primer was hybridised to the PCR product and extended using fluorescent terminating dideoxynucleotides which were detected by colorimetric assay. Using this ELISA-based assay, the prevalence of the factor V Leiden mutation was determined in an Australian blood donor population (n = 500). A total of 18 heterozygotes were identified (3.6%) and all of these were confirmed with conventional MnlI restriction digest. No homozygotes for the variant allele were detected. We conclude from this study that the frequency of 3.6% is compatible with others published for Caucasian populations. In addition, the FNC technology shows promise as the basis for a rapid, automated DNA based test for factor V Leiden.

Multiple analysis of three common genetic alterations associated with thrombophilia

We have previously reported the use of a novel mini-sequencing protocol for detection of the factor V Leiden variant, the first nucleotide change (FNC) technology. This technology is based on a single nucleotide extension of a primer, which is hybridized immediately adjacent to the site of mutation. The extended nucleotide that carries a reporter molecule (fluorescein) has the power to discriminate the genotype at the site of mutation. More recently, the prothrombin 20210 and thermolabile methylene tetrahydrofolate reductase (MTHFR) 677 variants have been identified as possible risk factors associated with thrombophilia. This study describes the use of the FNC technology in a combined assay to detect factor V, prothrombin and MTHFR variants in a population of Australian blood donors, and describes the objective numerical methodology used to determine genotype cut-off values for each genetic variation. Using FNC to test 500 normal blood donors, the incidence of Factor V Leiden was 3.6% (all heterozygous), that of prothrombin 20210 was 2.8% (all heterozygous) and that of MTHFR was 10% (homozygous). The combined FNC technology offers a simple, rapid, automatable DNA-based test for the detection of these three important mutations that are associated with familial thrombophilia. (C) 2000 Lippincott Williams and Wilkins.

Starting out in STEM : a study of young men and women in first year science, technology, engineering and mathematics courses

In late 2011, first year university students in science, technology, engineering and mathematics (STEM) courses across Australia were invited to participate in the international Interests and Recruitment in Science (IRIS) study. IRIS investigates the influences on young people's decisions to choose university STEM courses and their subsequent experiences of these courses. The study also has a particular focus on the motivations and experiences of young women in courses such as physics, IT and engineering given the low rates of female participation in these fields. Around 3500 students from 30 Australian universities contributed their views on the relative importance of various school and non-school influences on their decisions, as well as insights into their experiences of university STEM courses so far. It is hoped that their contributions will help improve recruitment, retention and gender equity in STEM higher education and careers.

Starting out in STEM : a study of young men and women in first year science, technology, engineering and mathematics courses

In late 2011, first year university students in science, technology, engineering and mathematics (STEM) courses across Australia were invited to participate in the international Interests and Recruitment in Science (IRIS) study. IRIS investigates the influences on young people's decisions to choose university STEM courses and their subsequent experiences of these courses. The study also has a particular focus on the motivations and experiences of young women in courses such as physics, IT and engineering given the low rates of female participation in these fields. Around 3500 students from 30 Australian universities contributed their views on the relative importance of various school and non-school influences on their decisions, as well as insights into their experiences of university STEM courses so far. It is hoped that their contributions will help improve recruitment, retention and gender equity in STEM higher education and careers.

Specificities of Caenorhabditis elegans and human hairpin binding proteins for the first nucleotide in the histone mRNA hairpin loop.

The 3' ends of animal replication-dependent histone mRNAs are formed by endonucleolytic cleavage of the primary transcripts downstream of a highly conserved RNA hairpin. The hairpin-binding protein (HBP) binds to this RNA element and is involved in histone RNA 3' processing. A minimal RNA-binding domain (RBD) of approximately 73 amino acids that has no similarity with other known RNA-binding motifs was identified in human HBP [Wang Z-F et al., Genes & Dev, 1996, 10:3028-3040]. The primary sequence identity between human and Caenorhabditis elegans RBDs is 55% compared to 38% for the full-length proteins. We analyzed whether differences between C. elegans and human HBP and hairpins are reflected in the specificity of RNA binding. The C. elegans HBP and its RBD recognize only their cognate RNA hairpins, whereas the human HBP or RBD can bind both the mammalian and the C. elegans hairpins. This selectivity of C. elegans HBP is mostly mediated by the first nucleotide in the loop, which is C in C. elegans and U in all other metazoans. By converting amino acids in the human RBD to the corresponding C. elegans residues at places where the latter deviates from the consensus, we could identify two amino acid segments that contribute to selectivity for the first nucleotide of the hairpin loop.

Climate change technology transfer to developing countries: evidence analysis and policy recommendations

Developing countries are experiencing unprecedented levels of economic growth. As a result, they will be responsible for most of the future growth in energy demand and greenhouse gas (GHG) emissions. Curbing GHG emissions in developing countries has become one of the cornerstones of a future international agreement under the United Nations Framework Convention for Climate Change (UNFCCC). However, setting caps for developing countries’ GHG emissions has encountered strong resistance in the current round of negotiations. Continued economic growth that allows poverty eradication is still the main priority for most developing countries, and caps are perceived as a constraint to future growth prospects. The development, transfer and use of low-carbon technologies have more positive connotations, and are seen as the potential path towards low-carbon development. So far, the success of the UNFCCC process in improving the levels of technology transfer (TT) to developing countries has been limited. This thesis analyses the causes for such limited success and seeks to improve on the understanding about what constitutes TT in the field of climate change, establish the factors that enable them in developing countries and determine which policies could be implemented to reinforce these factors. Despite the wide recognition of the importance of technology and knowledge transfer to developing countries in the climate change mitigation policy agenda, this issue has not received sufficient attention in academic research. Current definitions of climate change TT barely take into account the perspective of actors involved in actual climate change TT activities, while respective measurements do not bear in mind the diversity of channels through which these happen and the outputs and effects that they convey. Furthermore, the enabling factors for TT in non-BRIC (Brazil, Russia, India, China) developing countries have been seldom investigated, and policy recommendations to improve the level and quality of TTs to developing countries have not been adapted to the specific needs of highly heterogeneous countries, commonly denominated as “developing countries”. This thesis contributes to enriching the climate change TT debate from the perspective of a smaller emerging economy (Chile) and by undertaking a quantitative analysis of enabling factors for TT in a large sample of developing countries. Two methodological approaches are used to study climate change TT: comparative case study analysis and quantitative analysis. Comparative case studies analyse TT processes in ten cases based in Chile, all of which share the same economic, technological and policy frameworks, thus enabling us to draw conclusions on the enabling factors and obstacles operating in TT processes. The quantitative analysis uses three methodologies – principal component analysis, multiple regression analysis and cluster analysis – to assess the performance of developing countries in a number of enabling factors and the relationship between these factors and indicators of TT, as well as to create groups of developing countries with similar performances. The findings of this thesis are structured to provide responses to four main research questions: What constitutes technology transfer and how does it happen? Is it possible to measure technology transfer, and what are the main challenges in doing so? Which factors enable climate change technology transfer to developing countries? And how do different developing countries perform in these enabling factors, and how can differentiated policy priorities be defined accordingly? vi Resumen Los paises en desarrollo estan experimentando niveles de crecimiento economico sin precedentes. Como consecuencia, se espera que sean responsables de la mayor parte del futuro crecimiento global en demanda energetica y emisiones de Gases de Efecto de Invernadero (GEI). Reducir las emisiones de GEI en los paises en desarrollo es por tanto uno de los pilares de un futuro acuerdo internacional en el marco de la Convencion Marco de las Naciones Unidas para el Cambio Climatico (UNFCCC). La posibilidad de compromisos vinculantes de reduccion de emisiones de GEI ha sido rechazada por los paises en desarrollo, que perciben estos limites como frenos a su desarrollo economico y a su prioridad principal de erradicacion de la pobreza. El desarrollo, transferencia y uso de tecnologias bajas en carbono tiene connotaciones mas positivas y se percibe como la via hacia un crecimiento bajo en carbono. Hasta el momento, la UNFCCC ha tenido un exito limitado en la promocion de transferencias de tecnologia (TT) a paises en desarrollo. Esta tesis analiza las causas de este resultado y busca mejorar la comprension sobre que constituye transferencia de tecnologia en el area de cambio climatico, cuales son los factores que la facilitan en paises en desarrollo y que politicas podrian implementarse para reforzar dichos factores. A pesar del extendido reconocimiento sobre la importancia de la transferencia de tecnologia a paises en desarrollo en la agenda politica de cambio climatico, esta cuestion no ha sido suficientemente atendida por la investigacion existente. Las definiciones actuales de transferencia de tecnologia relacionada con la mitigacion del cambio climatico no tienen en cuenta la diversidad de canales por las que se manifiestan o los efectos que consiguen. Los factores facilitadores de TT en paises en desarrollo no BRIC (Brasil, Rusia, India y China) apenas han sido investigados, y las recomendaciones politicas para aumentar el nivel y la calidad de la TT no se han adaptado a las necesidades especificas de paises muy heterogeneos aglutinados bajo el denominado grupo de "paises en desarrollo". Esta tesis contribuye a enriquecer el debate sobre la TT de cambio climatico con la perspectiva de una economia emergente de pequeno tamano (Chile) y el analisis cuantitativo de factores que facilitan la TT en una amplia muestra de paises en desarrollo. Se utilizan dos metodologias para el estudio de la TT a paises en desarrollo: analisis comparativo de casos de estudio y analisis cuantitativo basado en metodos multivariantes. Los casos de estudio analizan procesos de TT en diez casos basados en Chile, para derivar conclusiones sobre los factores que facilitan u obstaculizan el proceso de transferencia. El analisis cuantitativo multivariante utiliza tres metodologias: regresion multiple, analisis de componentes principales y analisis cluster. Con dichas metodologias se busca analizar el posicionamiento de diversos paises en cuanto a factores que facilitan la TT; las relaciones entre dichos factores e indicadores de transferencia tecnologica; y crear grupos de paises con caracteristicas similares que podrian beneficiarse de politicas similares para la promocion de la transferencia de tecnologia. Los resultados de la tesis se estructuran en torno a cuatro preguntas de investigacion: .Que es la transferencia de tecnologia y como ocurre?; .Es posible medir la transferencia de tecnologias de bajo carbono?; .Que factores facilitan la transferencia de tecnologias de bajo carbono a paises en desarrollo? y .Como se puede agrupar a los paises en desarrollo en funcion de sus necesidades politicas para la promocion de la transferencia de tecnologias de bajo carbono?

Department of Energy : proposed budget in support of the President's Climate Change Technology Initiative : report to the Chairman, Committee on the Budget, House of Representatives /

"B-279612"--P. 1.

Utilizing Traditional Cognitive Measures of Academic Preparation to Predict First-Year Science, Technology, Engineering, and Mathematics (STEM) Majors' Success in Math and Science Courses

For the past several years, U.S. colleges and universities have faced increased pressure to improve retention and graduation rates. At the same time, educational institutions have placed a greater emphasis on the importance of enrolling more students in STEM (science, technology, engineering and mathematics) programs and producing more STEM graduates. The resulting problem faced by educators involves finding new ways to support the success of STEM majors, regardless of their pre-college academic preparation. The purpose of my research study involved utilizing first-year STEM majors’ math SAT scores, unweighted high school GPA, math placement test scores, and the highest level of math taken in high school to develop models for predicting those who were likely to pass their first math and science courses. In doing so, the study aimed to provide a strategy to address the challenge of improving the passing rates of those first-year students attempting STEM-related courses. The study sample included 1018 first-year STEM majors who had entered the same large, public, urban, Hispanic-serving, research university in the Southeastern U.S. between 2010 and 2012. The research design involved the use of hierarchical logistic regression to determine the significance of utilizing the four independent variables to develop models for predicting success in math and science. The resulting data indicated that the overall model of predictors (which included all four predictor variables) was statistically significant for predicting those students who passed their first math course and for predicting those students who passed their first science course. Individually, all four predictor variables were found to be statistically significant for predicting those who had passed math, with the unweighted high school GPA and the highest math taken in high school accounting for the largest amount of unique variance. Those two variables also improved the regression model’s percentage of correctly predicting that dependent variable. The only variable that was found to be statistically significant for predicting those who had passed science was the students’ unweighted high school GPA. Overall, the results of my study have been offered as my contribution to the literature on predicting first-year student success, especially within the STEM disciplines.

Rapid detection of SMARCB1 sequence variation using high resolution melting

Background : Rhabdoid tumors are rare cancers of early childhood arising in the kidney, central nervous system and other organs. The majority are caused by somatic inactivating mutations or deletions affecting the tumor suppressor locus SMARCB1 [OMIM 601607]. Germ-line SMARCB1 inactivation has been reported in association with rhabdoid tumor, epitheloid sarcoma and familial schwannomatosis, underscoring the importance of accurate mutation screening to ascertain recurrence and transmission risks. We describe a rapid and sensitive diagnostic screening method, using high resolution melting (HRM), for detecting sequence variations in SMARCB1. Methods : Amplicons, encompassing the nine coding exons of SMARCB1, flanking splice site sequences and the 5' and 3' UTR, were screened by both HRM and direct DNA sequencing to establish the reliability of HRM as a primary mutation screening tool. Reaction conditions were optimized with commercially available HRM mixes. Results : The false negative rate for detecting sequence variants by HRM in our sample series was zero. Nine amplicons out of a total of 140 (6.4%) showed variant melt profiles that were subsequently shown to be false positive. Overall nine distinct pathogenic SMARCB1 mutations were identified in a total of 19 possible rhabdoid tumors. Two tumors had two distinct mutations and two harbored SMARCB1 deletion. Other mutations were nonsense or frame-shifts. The detection sensitivity of the HRM screening method was influenced by both sequence context and specific nucleotide change and varied from 1: 4 to 1:1000 (variant to wild-type DNA). A novel method involving digital HRM, followed by re-sequencing, was used to confirm mutations in tumor specimens containing associated normal tissue. Conclusions : This is the first report describing SMARCB1 mutation screening using HRM. HRM is a rapid, sensitive and inexpensive screening technology that is likely to be widely adopted in diagnostic laboratories to facilitate whole gene mutation screening.

Development of an oligonucleotide-based SNP detection method on lateral flow strips using hexapet tages

Detection of point mutations or single nucleotide polymorphisms (SNPs) is important in relation to disease susceptibility or detection in pathogens of mutations determining drug resistance or host range. There is an emergent need for rapid detection methods amenable to point-of-care applications. The purpose of this study was to reduce to practice a novel method for SNP detection and to demonstrate that this technology can be used downstream of nucleic acid amplification. The authors used a model system to develop an oligonucleotide-based SNP detection system on nitrocellulose lateral flow strips. To optimize the assay they used cloned sequences of the herpes simplex virus-1 (HSV-1) DNA polymerase gene into which they introduced a point mutation. The assay system uses chimeric polymerase chain reaction (PCR) primers that incorporate hexameric repeat tags ("hexapet tags"). The chimeric sequences allow capture of amplified products to predefined positions on a lateral flow strip. These "hexapet" sequences have minimal cross-reactivity and allow specific hybridization-based capture of the PCR products at room temperature onto lateral flow strips that have been striped with complementary hexapet tags. The allele-specific amplification was carried out with both mutant and wild-type primer sets present in the PCR mix ("competitive" format). The resulting PCR products carried a hexapet tag that corresponded with either a wild-type or mutant sequence. The lateral flow strips are dropped into the PCR reaction tube, and mutant sequence and wild-type sequences diffuse along the strip and are captured at the corresponding position on the strip. A red line indicative of a positive reaction is visible after 1 minute. Unlike other systems that require separate reactions and strips for each target sequence, this system allows multiplex PCR reactions and multiplex detection on a single strip or other suitable substrates. Unambiguous visual discrimination of a point mutation under room temperature hybridization conditions was achieved with this model system in 10 minutes after PCR. The authors have developed a capture-based hybridization method for the detection and discrimination of HSV-1 DNA polymerase genes that contain a single nucleotide change. It has been demonstrated that the hexapet oligonucleotides can be adapted for hybridization on the lateral flow strip platform for discrimination of SNPs. This is the first step in demonstrating SNP detection on lateral flow using the hexapet oligonucleotide capture system. It is anticipated that this novel system can be widely used in point-of-care settings.

Managing Strategic Change

The essay - Managing Strategic Change – by K. Michael Haywood, Associate Professor, School of Hotel and Food Administration, University of Guelph, is initially characterized by Haywood as: “The ability to manage strategic change is critical for hospitality industry executives today. Executives must be capable of creating a vision of the future and implementing its direction. The author gives avenues for that management process.” “The effective management of strategic change is the major challenge confronting hospitality executives,” says Associate Professor Haywood. “Responding to a rapidly changing business environment and constantly evolving competitive threats and opportunities requires executives who can anticipate and plan for change.” According to Professor Haywood, the management of strategic change is a future imperative for hospitality executives. Implementing those changes will be even more difficult. “Survival and growth for many hospitality firms during the next decade will depend on the development of new strategic visions which can provide significant competitive advantages,” he says. “Strategies for managing costs and technology will be central to this task,” Haywood expands the thought. Haywood suggests two primary types of change hospitality executives should be aware of. First, is change that is anticipated, anticipatory change. Second, is the other more crucial type of change, strategic change in the face of crisis, or simply stated, reactive change. Professor Haywood describes the distinction between the two. In describing the approach that should be implemented in responding to an anticipatory change, Haywood says, “If time permits, and change is to be introduced gradually, pilots and trials should be run to assess the impact of the new strategy on the organization. These trials are used to create pockets of commitment throughout the corporation, build comfort levels with the new approach, and neutralize or win over potential opposition.” There are the obvious advantages to using an approach like the one described above, but there are disadvantages as well. Haywood discusses both. In addressing reactive change, Haywood offers that the process is a more - time is of the essence – condition, and that strong leadership and a firm hand on employee control is imperative. “Personal leadership, tough-mindedness, the willingness to ruthlessly abandon the familiar and the past, and the use of informal strategic levers are the hallmarks of sterling executive performance in such periods,” he says. “All these changes involve substantial technical, financial, and human risks,” Haywood wants you to know. “In order to make them, and still remain competitive, hospitality and travel-related corporations require executives capable of creating a vision of the future, able to sell that vision to their employees, and tough-minded enough to implement strategies to make the vision a reality.”

Make-buy decisions in the face of technological change: does industry clockspeed matter?

Some evidence in the area of make-buy decisions for new technologies suggests that it is a good idea for a company to pursue a fairly rigorous ''make'' policy in the early days of a potentially disruptive innovation. Other studies prescribe exactly the opposite, promoting instead a ''buy'' strategy. This paper seeks to bridge the gap between these perspectives by suggesting that both strategies are valid, but that they are most successfully applied in different market environments. The ''make'' prescription may be more suited to either extremely fast or extremely slow rates of technological change, while a ''buy'' strategy might be more appropriate in market sectors where technologies evolve at a medium pace. This paper highlights the importance of industry clockspeed and supplier relationships in make-buy decisions for new technologies, and puts forward two new hypotheses that require empirical testing.

In arrayed ranks array technology in the study of mesothelioma steven

Mesothelioma is a rare malignancy arising from mesothelial cells lining the pleura and peritoneum. Advances in modern technology have allowed the development of array based approaches to the study of disease allowing researchers the opportunity to study many genes or proteins in a high-throughput fashion. This review describes the current knowledge surrounding array based approaches with respect to mesothelioma research. © 2009 by the International Association for the Study of Lung Cancer.