Cloning and characterisation of two subtilisin-like protease genes from Neotyphodium lolii : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Molecular Genetics at Massey University

PCR amplification of Neotyphodium lolii genomic DNA with degenerate primers detected two different sequences with homology to subtilisin-like proteases. These two PCR products were used to screen a N. lolii Lp19 genomic library. The prt1 gene was isolated by screening the genomic library with the GH30 PCR product. This gene encodes a putative peptide of 434 amino acids that is most similar to subtilisin-like proteases from Aspergillus sp. The prt1 gene contained a single intron, which was in a position conserved with other fungal genes. 3'RACE was used to determine the polyadenylation site for the prt1 gene. Repetitive DNA was a feature of both the 3' untranslated region (UTR) and sequences downstream of the prtl gene. Within the 3' UTR, a complex microsatellite was found extending over 50 base pairs. Downstream of the gene, a minisatellite locus of 360 base pairs in size was found, consisting of 40 copies of a 9 base pair AT-rich repeat. Expression of prt1 was examined in cultures with various types of carbon and nitrogen sources. Although no conclusive results could be drawn, the type of carbon and nitrogen available did have some effect on prt1 expression. Repression of prt1 expression was only observed in media supplemented with sucrose and glutamate. A 500 bp fragment from the prt1 promoter was introduced into the vector pFunGus to create a translational fusion with gusA. This vector, pMM9, was transformed into Penicillium paxilli. Although transformation frequencies were low, the transformants obtained appeared to be stable for hygromycin resistance. Expression of GUS was observed in seven out of twelve of the stable transformants. This showed that the promoter fragment in pMM9 was sufficient for expression of GUS in a heterologous system. The prt2 gene was isolated by screening a genomic library with the GH3 PCR product. Partial sequence has been obtained for the prt2 gene. The prt2 gene contains at least three introns, the first of which is conserved with prt1. From the sequence obtained, prt2 encodes a peptide with strong similarity to subtilisin-like proteases from Metarhizium anisopliae, a fungal pathogen of insects

Senior Woman Administrator: The Definition, Challenges, Influence, and Perceptions

Diversity efforts implemented by the National Collegiate Athletic Association (NCAA) hope to improve the Association through the addition of multiple voices in athletics. Notably, the Senior Woman Administrator (SWA) designation is intended to encourage and promote the involvement of female administrators in meaningful ways in the decision-making process in intercollegiate athletics. This role, created under Article 4.02.4 of the NCAA constitution, is to be filled by the highest ranking female in each NCAA athletic department or member conference (Levick, 2002; Raphaely, 2003). Given the evolving definition and nature of the SWA designation and of female managerial roles (Eagly & Karau, 2002), there exists a question as to whether the SWA designation has provided the scope of decision-making and authority suggested in the NCAA definition of the designation. Research must show if SWAs are able to use their power and give different opinions. It must also uncover if there is still a need for the SWA role and if the title is still appropriate for this designation

Who does what now? How physics lab instruction impacts student behaviors

While laboratory instruction is a cornerstone of physics education, the impact of student behaviours in labs on retention, persistence in the field, and the formation of students' physics identity remains an open question. In this study, we performed in-lab observations of student actions over two semesters in two pedagogically different sections of the same introductory physics course. We used a cluster analysis to identify different categories of student behaviour and analyzed how they correlate with lab structure and gender. We find that, in lab structures which fostered collaborative group work and promoted decision making, there was a task division along gender lines with respect to laptop and equipment usage (and found no such divide among students in guided verification labs).Comment: 4 pages, 3 figures, 3 table

Determination of Calories in Food Via Adiabatic Bomb Calorimeter

The adiabatic bomb calorimeter has been an effective tool in facilitating heat transfer between molecules via combustion reaction. Heat is released from the substance combusted and transferred into another, usually water. The heat transferred is measured and the enthalpy of combustion determined for the combusted material. One such classic experiment is the determination of combustion enthalpy of sucrose, obtaining the calorimeter constant with benzoic acid. The results of an adiabatic combustion experiment can be taken one step further to determine the combustion enthalpy in kcal/g by a simple conversion calculation. Since food nutrition labels report calories per grams of serving (note: 1 food calorie 1 kcal), food can be combusted in an adiabatic bomb calorimeter and the results compared to its corresponding nutrition label. The food chosen for this experiment consisted of marshmallows and cheddar cheese

Increased TNF-α/IFN-γ/IL-2 and Decreased TNF-α/IFN-γ Production by Central Memory T Cells Are Associated with Protective Responses against Bovine Tuberculosis Following BCG Vaccination

Central memory T cells (Tcm) and polyfunctional CD4 T cell responses contribute to vaccine-elicited protection with both human and bovine tuberculosis (TB); however, their combined role in protective immunity to TB is unclear. To address this question, we evaluated polyfunctional cytokine responses by CD4 T cell effector / memory populations from bacille Calmette Guerin (BCG) vaccinated and non-vaccinated calves prior to and after aerosol challenge with virulent Mycobacterium bovis. Polyfunctional cytokine expression patterns in the response by Tcm, effector memory, and effector T cell subsets were similar between BCG-vaccinated and M. bovis-infected calves; only differing in magnitude (i.e., infected > vaccinated). BCG vaccination, however, did alter the kinetics of the ensuing response to virulent M. bovis infection. Early after challenge (three weeks post-infection), non-vaccinates had greater antigen-specific IFN-γ/TNF-α and lesser IFN-γ/TNF-α/IL-2 responses by Tcm cells than did vaccinated animals. Importantly, these differences were also associated with mycobacterial burden upon necropsy. Polyfunctional responses to ESAT-6:CFP10 (antigens not synthesized by BCG strains) were detected in memory subsets, as well as in effector cells, as early as three weeks after challenge. These findings suggest that cell fate divergence may occur early after antigen priming in the response to bovine TB and that memory and effector T cells may expand concurrently during the initial phase of the immune response. In summary, robust IFN-γ/TNF-α response by Tcm cells is associated with greater mycobacterial burden while IFN-γ/TNF-α/IL-2 response by Tcm cells are indicative of a protective response to bovine TB

A Social Model for Health Promotion for an Aging Population: Initial Evidence on the Experience Corps Model

This report evaluates whether a program for older volunteers, designed for both generativity and health promotion, leads to short-term improvements in multiple behavioral risk factors and positive effects on intermediary risk factors for disability and other morbidities. The Experience Corps® places older volunteers in public elementary schools in roles designed to meet schools\u27 needs and increase the social, physical, and cognitive activity of the volunteers. This article reports on a pilot randomized trial in Baltimore, Maryland. The 128 volunteers were 60-86 years old; 95% were African American. At follow-up of 4-8 months, physical activity, strength, people one could turn to for help, and cognitive activity increased significantly, and walking speed decreased significantly less, in participants compared to controls. In this pilot trial, physical, cognitive, and social activity increased, suggesting the potential for the Experience Corps to improve health for an aging population and simultaneously improve educational outcomes for children

Application of the Saffir-Simpson Hurricane Wind Scale to Assess Sand Dune Response to Tropical Storms

Over one-third of the Earth’s population resides or works within 200 km of the coast. The increasing threat of coastal hazards with predicted climate change will impact many global citizens. Coastal dune systems serve as a natural first line of defense against rising sea levels and coastal storms. This study investigated the volumetric changes of two dune systems on Isle of Palms, South Carolina, USA prior to and following Hurricanes Irma (2017) and Florence (2018), which impacted the island as tropical storms with different characteristics. Irma had relatively high significant wave heights and precipitation, resulting in an average 39% volumetric dune loss. During Florence, a storm where precipitation was low and winds were moderate, net volumetric dune loss averaged 3%. The primary driving force causing dune change during Irma was water (precipitation and storm surge), and during Florence, it was wind (aeolian transport). We suggest that the application of the Saffir-Simpson Hurricane Wind Scale classifications should be reconsidered because different geomorphic responses were measured, despite Irma and Florence both being designated as tropical storms. Site-specific pre- and post-storm studies of the dune morphology and site-specific meteorological measurements of the storm (wind characteristics, storm surge, precipitation) are critically needed

Impact of Experience Corps® Participation on Children’s Academic Achievement and School Behavior

This article reports on the impact of the Experience Corps® (EC) Baltimore program, an intergenerational, school-based program aimed at improving academic achievement and reducing disruptive school behavior in urban, elementary school students in Kindergarten through third grade (K-3). Teams of adult volunteers aged 60 and older were placed in public schools, serving 15 h or more per week, to perform meaningful and important roles to improve the educational outcomes of children and the health and well-being of volunteers. Findings indicate no significant impact of the EC program on standardized reading or mathematical achievement test scores among children in grades 1–3 exposed to the program. K-1st grade students in EC schools had fewer principal office referrals compared to K-1st grade students in matched control schools during their second year in the EC program; second graders in EC schools had fewer suspensions and expulsions than second graders in non-EC schools during their first year in the EC program. In general, both boys and girls appeared to benefit from the EC program in school behavior. The results suggest that a volunteer engagement program for older adults can be modestly effective for improving selective aspects of classroom behavior among elementary school students in under-resourced, urban schools, but there were no significant improvements in academic achievement. More work is needed to identify individual- and school-level factors that may help account for these results