Master of Science

thesisGaucher disease (GD) is an autosomal recessive disorder caused by deficiency of the lysosomal hydrolase glucocerebrosidase (GBA) (EC 3.2.1.45), required for the degradation of glycosphingolipids. In Gaucher disease, as a result of GBA deficiency, the body is unable to breakdown glucocerebroside, thus allowing the accumulation of glucosylceramide and glucosphingosine within the lysosomal cells. The clinical presentation of GD consists in a multisystem disorder, and its classification of GD into its three subtypes is dependent primarily on the presence or absence of neurological features. Gaucher disease type 1 is the most common inherited genetic disease affecting Ashkenazi Jews, with a frequency of approximately 1 in 850 individuals having Gaucher disease and 1 in 18 individuals being carriers. Once Gaucher disease is suspected based on clinical presentation, the first line of testing is measuring the glucocerebrosidase (GBA) enzyme activity. The GBA enzymatic assay has been developed as a part of this study by modifying the conditions described by Urban et al (2008). The process of validation was divided into individual steps identified to provide the most optimum conditions for the implementation of this assay. This assay uses the fluorometric methodology to measure the level of enzymatic activity. The fluorometric signal is directly proportional to the concentration of GBA activity in the individual being assessed. iv The evaluation of all components of the method validation included accuracy, precision, linearity, sample requirements, sample stability, and establishment of reference ranges. After the individual assessment of these components, it was concluded that the assay was successfully validated for the measurement of GBA activity, and it provides a valuable tool as the first line of testing for the diagnosis of Gaucher disease

Rejuvenation by cell reprogramming: A new horizon in gerontology

The discovery of animal cloning and subsequent development of cell reprogramming technology were quantum leaps as they led to the achievement of rejuvenation by cell reprogramming and the emerging view that aging is a reversible epigenetic process. Here, we will first summarize the experimental achievements over the last 7 years in cell and animal rejuvenation. Then, a comparison will be made between the principles of the cumulative DNA damage theory of aging and the basic facts underlying the epigenetic model of aging, including Horvath's epigenetic clock. The third part will apply both models to two natural processes, namely, the setting of the aging clock in the mammalian zygote and the changes in the aging clock along successive generations in mammals. The first study demonstrating that skin fibroblasts from healthy centenarians can be rejuvenated by cell reprogramming was published in 2011 and will be discussed in some detail. Other cell rejuvenation studies in old humans and rodents published afterwards will be very briefly mentioned. The only in vivo study reporting that a number of organs of old progeric mice can be rejuvenated by cyclic partial reprogramming will also be described in some detail. The cumulative DNA damage theory of aging postulates that as an animal ages, toxic reactive oxygen species generated as byproducts of the mitochondria during respiration induce a random and progressive damage in genes thus leading cells to a progressive functional decline. The epigenetic model of aging postulates that there are epigenetic marks of aging that increase with age, leading to a progressive derepression of DNA which in turn causes deregulated expression of genes that disrupt cell function. The cumulative DNA damage model of aging fails to explain the resetting of the aging clock at the time of conception as well as the continued vitality of species as millenia go by. In contrast, the epigenetic model of aging straightforwardly explains both biologic phenomena. A plausible initial application of rejuvenation in vivo would be preventing adult individuals from aging thus eliminating a major risk factor for end of life pathologies. Further, it may allow the gradual achievement of whole body rejuvenation.Fil: Goya, Rodolfo Gustavo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Lehmann, Marianne. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Chiavellini, Priscila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Canatelli Mallat, Martina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; ArgentinaFil: Hereñú, Claudia Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Farmacología Experimental de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Instituto de Farmacología Experimental de Córdoba; ArgentinaFil: Brown, Oscar Alfredo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner". Universidad Nacional de la Plata. Facultad de Ciencias Médicas. Instituto de Investigaciones Bioquímicas de La Plata "Prof. Dr. Rodolfo R. Brenner"; Argentin

VIBCO Bin Variation System

VIBCO vibrators has tasked the team in designing an internal bin fixture that aids in material flow for the food, pharmaceutical, and chemical industries. This bin fixture must be adaptable to various sized diameter bins in order to create a cost efficient product that does not require the customer to make any physical changes to their bin. Lastly the internal fixture must be capable of raising and lowering in a way that gives the customer the ability to control material flow. The team began research on competitor systems that are currently on the market in this industry. After completing this research, it was determined that all of the competitor systems require the customer to make physical changes to their bin in order to implement the system. This being said it was imperative that the team design an internal fixture that can be easily installed without requiring the customer to edit their bin. From here design specifications were determined and discussed with the VIBCO engineering team to ensure that all of the bases were covered. In order to complete this task, the team began with brainstorming in order to determine some basic design specifications that would guide our 90 concepts generated. After analyzing each of the 90 concepts and a critical design review, two concepts were created by integrating and incorporating many elements of the entire spectrum of concept solutions. From here the team furthered research on different materials that would be strong, light weight, cost efficient, and high-corrosive resistant. After determining the different materials design work was furthered and a Solidworks design of the full internal bin system was created. Before finalizing any Solidworks designs the team crosschecked once again the proposed design with the competitors to ensure that this had not been done before. After completing and simulating a model of the proposed design the team performed a financial analysis and determined we are well under competitor pricing once this design is in full production. All of our other design specifications were met but the team would continue to make improvements. Once the design had been approved by VIBCO, parts were ordered and the build portion of the project began. Testing of the system soon followed at VIBCO using a bin they had on site. With the testing and data collected, redesign and improvements were able to be made to the original prototype

CHOICES: A Family-based Childhood Obesity Intervention for Low Income Minority Children

The objective of this presentation is to discuss the effectiveness of a 6-week summer childhood obesity intervention for low-income minority children ages 10-12. Topics will include strategies and best practices for effective summer programming for youth. The target audience includes practitioners, educators, and researchers interested in childhood obesity interventions involving families and communities

The N-terminal PIN domain of the exosome subunit Rrp44 harbors endonuclease activity and tethers Rrp44 to the yeast core exosome

Nuclear and cytoplasmic forms of the yeast exosome share 10 components, of which only Rrp44/Dis3 is believed to possess 3′ exonuclease activity. We report that expression only of Rrp44 lacking 3′-exonuclease activity (Rrp44-exo) supports growth in S288c-related strains (BY4741). In BY4741, rrp44-exo was synthetic-lethal with loss of the cytoplasmic 5′-exonuclease Xrn1, indicating block of mRNA turnover, but not with loss of the nuclear 3′-exonuclease Rrp6. The RNA processing phenotype of rrp44-exo was milder than that seen on Rrp44 depletion, indicating that Rrp44-exo retains important functions. Recombinant Rrp44 was shown to possess manganese-dependent endonuclease activity in vitro that was abolished by four point mutations in the putative metal binding residues of its N-terminal PIN domain. Rrp44 lacking both exonuclease and endonuclease activity failed to support growth in strains depleted of endogenous Rrp44. Strains expressing Rrp44-exo and Rrp44-endo–exo exhibited different RNA processing patterns in vivo suggesting Rrp44-dependent endonucleolytic cleavages in the 5′-ETS and ITS2 regions of the pre-rRNA. Finally, the N-terminal PIN domain was shown to be necessary and sufficient for association with the core exosome, indicating its dual function as a nuclease and structural element

Professional development of teachers at Malta’s Giovanni Curmi Higher Secondary School : contributions from a stake-holding think tank

This paper synthesises a group of educators’ engagement with an action research project endorsed by the Council of Europe’s Pestalozzi Programme’s Action Research Group 2013 – 2014. Educators fulfilling duties at Giovanni Curmi Higher Secondary School (GCHSS) collaborated within the context of a think-tank by engaging in action research on professional development for teachers at Giovanni Curmi Higher Secondary School (PDGCHSS). Data that emerged from the foregoing research indicate that professional development (PD) should be grounded in, and not disconnected from, the school-context, and must be relevant to the teachers’ everyday teaching. In addition, knowledge gained from PD training should not be sporadic or disconnected, but transferable to everyday practice. A highly bureaucratized system and lack of resources, such as time, financial resources and technological skills, are detrimental to teachers’ motivation for PDpeer-reviewe

Quantitative sensory testing in children with sickle cell disease: additional insights and future possibilities.

Quantitative sensory testing (QST) is used in a variety of pain disorders to characterize pain and predict prognosis and response to specific therapies. In this study, we aimed to confirm results in the literature documenting altered QST thresholds in sickle cell disease (SCD) and assess the test-retest reliability of results over time. Fifty-seven SCD and 60 control subjects aged 8-20 years underwent heat and cold detection and pain threshold testing using a Medoc TSAII. Participants were tested at baseline and 3 months; SCD subjects were additionally tested at 6 months. An important facet of our study was the development and use of a novel QST modelling approach, allowing us to model all data together across modalities. We have not demonstrated significant differences in thermal thresholds between subjects with SCD and controls. Thermal thresholds were consistent over a 3- to 6-month period. Subjects on whom hydroxycarbamide (HC) was initiated shortly before or after baseline testing (new HC users) exhibited progressive decreases in thermal sensitivity from baseline to 6 months, suggesting that thermal testing may be sensitive to effective therapy to prevent vasoocclusive pain. These findings inform the use of QST as an endpoint in the evaluation of preventative pain therapies