Lack of evidence for decreased protein stability in the 2397 (Met) haplotype of the leucine rich repeat kinase 2 protein implicated in Parkinson’s disease

Missense mutations in the leucine rich repeat kinase 2 (LRRK2) gene are the leading genetic cause of autosomal dominant familial Parkinson’s disease. We previously reported that two mutations within the ROC domain, namely R1441C and A1442P, exhibit increased protein degradation leading to lowered steady state LRRK2 protein levels in HEK293 cells. More recently, the common WD40 domain LRRK2 haplotype, Met2397, which is a risk factor for Crohn’s disease, has been shown to lower steady state protein levels in HEK293 cells. In view of recent evidence implicating LRRK2 and inflame-mation in PD, we investigated the effects of Met2397 on LRRK2 expression, and compared them to the Thr2397 variant and other LRRK2 mutants. In this study, we transfected HEK293 cells with plasmid constructs encoding the different LRRK2 variants, and analyzed the resulting protein levels by Western blot and flow cytometry. Here we found that both the Met2397 and Thr2397 haplotypes yield similar levels of LRRK2 protein expression and do not appear to impact cell viability in HEK293 cells, compared to other LRRK mutants. Thus, we have concluded that the Met2397 haplotype is unlikely to play a role in LRRK2 mediated or idiopathic PD

Bactericidal Effects of Cold Plasma Technology on Geobacillus Stearothermophilus and Bacillus Cereus Microorganisms

Cold plasma is a state of matter that contains a large number of particles that are electrically charged. Plasmas generate chemically reactive species and ultraviolet radiation making them useful in decontamination applications (Kong & Laroussi, 2003). Research regarding the inactivation of gram-positive bacteria by cold plasma has been studied by Laroussi et al (2003); however, there is limited research regarding the germicidal effectiveness of cold plasma on Geobacillus stearothermophilus and Bacillus cereus microorganisms. The purpose of this study was to determine if cold plasma technology inactivates Geobacillus stearothermophilus and Bacillus cereus vegetative cells and spores. This study consisted of 981 samples; 762 experimental samples exposed to cold plasma at various times and 291 controls. Experimental samples were inoculated and exposed either directly or indirectly/remotely to cold plasma. After exposure the samples were incubated for 12 to 16 hours and colony forming units (CFU) were quantified. The percentage kill and log concentration reductions were calculated from the CFU counts. Data was analyzed using one-way ANOVA, Kruskal Wallis and Tukey\u27s tests at the .05 level. There was a statistically significant difference in the inactivation of Geobacillus stearothermophilus vegetative cells for indirect exposure (p=.0001), direct exposure (p=.0013), as well as for Bacillus cereus vegetative cells and spores (p=.0001). Exposure of Geobacillus stearothermophilus spores to cold plasma demonstrated no statistically significant differences in inactivation for indirect exposure (p=.7208) and direct exposure (p=.0835). Results indicate that cold plasma exposure significantly inactivated Geobacillus stearothermophilus (vegetative) and Bacillus cereus; however, Geobacillus stearothermophilus spores were not significantly inactivated. Funding for this project was provided by ADHA IOH

Cold Plasma Technology: Bactericidal Effects on Geobacillus Stearothermophilus and Bacillus Cereus Microorganisms

Introduction: Cold plasma, also known as Low Temperature Atmospheric Pressure Plasma (LTAPP) is a novel technology consisting of neutral and charged particles, including free radicals, which can be used to destroy or inactivate microorganisms. Research has been conducted regarding the effect of cold plasma on gram-positive bacteria; however, there is limited research regarding its ability to inactivate the spore-formers Geobacillus stearothermophilus and Bacillus cereus. Purpose: The purpose of this study was to determine if cold plasma inactivates G. stearothermophilus and B. cereus vegetative cells and spores. Methods: Nine hundred eighty-one samples were included in this study (762 experimental and 219 controls). Experimental samples were exposed indirectly or directly to cold plasma, before plating and incubating for 16 hours. Control samples were not exposed to cold plasma. The percentage-kill and cell number reductions were calculated from Colony Forming Units (CFU). Data were statistically analyzed at the .05 level using one-way ANOVA, Kruskal Wallis and Tukey\u27s tests. Results: There was a statistically significant difference in the inactivation of G. stearothermophilus vegetative cells receiving indirect and direct exposure (p=0.0001 and p=0.0013, respectively), as well as for B. cereus vegetative cells and spores (p=0.0001 for direct and indirect). There was no statistically significant difference in the inactivation of G. stearothermophilus spores receiving indirect exposure (p=0.7208) or direct exposure (p=0.0835). Conclusion: Results demonstrate that cold plasma exposure effectively kills G. stearothermophilus vegetative cells and B. cereus vegetative cells and spores; however, G. stearothermophilus spores were not significantly inactivated

The barriers to and facilitators of physical activity and sport for Oceania with Non-European, Non-Asian (ONENA) ancestry children and adolescents : A mixed studies systematic review

Background: Participation in sport and physical activity (PA) leads to better overall health, increased life expectancy, and decreased mortality rates across the lifespan; however, there may be a range of individual, family, and community factors that influence PA participation among ONENA children and adolescents residing in the 22 Pacific Island Countries and Territories (PICT) and Australia. This review aimed to synthesise existing quantitative and qualitative literature regarding barriers to and facilitators of PA and sport among ONENA youth. Methods: The literature was systematically searched to include studies reporting barriers to and facilitators of PA and sports participation among ONENA children and adolescents aged 0–18 years residing in the 22 PICT and Australia. Using a pre-established taxonomy based on the social-ecological model, a deductive analysis was performed. Quality appraisal was performed using the mixed methods appraisal tool. Results: Of 1388 articles, 14 studies were included, with 128 ONENA children and adolescent participants across the four qualitative studies; 156,581 ONENA children and adolescents across the seven quantitative studies; 801 parents, children, and adolescents in one quantitative study; and 642 parents in two quantitative studies. Of the 14 included studies, none were based in Australia and only 10 of the 22 PICT were reported as the participants’ residence: Palau, New Zealand, Tonga, Cook Islands, Kiribati, Samoa, Solomon Islands, Tuvalu, Vanuatu, and Fiji. Four studies reported barriers, and another four studies reported facilitators of PA and sport, with the remaining studies reporting both barriers and facilitators. Overall, there were more barriers reported (30 in total) than facilitators (27 in total). Conclusions: Research in this area is lacking, with ONENA youth living in Australia and 12 PICT not represented. Overall, there were a larger number of facilitators experienced at individual and interpersonal levels, while barriers were highest at the community level, with the policy level having facilitators and barriers equally represented. Programs that offer PA and sport participation options with embedded SDT-informed strategies for all family members; that are accessible through existing transport and related social, cultural, and physical infrastructure; and that are committed to communities through formal co-design partnerships are needed, to enhance the PA and sport participation of ONENA youth residing in PICT

Caffeine exposure induces browning features in adipose tissue in vitro and in vivo

Brown adipose tissue (BAT) is able to rapidly generate heat and metabolise macronutrients, such as glucose and lipids, through activation of mitochondrial uncoupling protein 1 (UCP1). Diet can modulate UCP1 function but the capacity of individual nutrients to promote the abundance and activity of UCP1 is not well established. Caffeine consumption has been associated with loss of body weight and increased energy expenditure, but whether it can activate UCP1 is unknown. This study examined the effect of caffeine on BAT thermogenesis in vitro and in vivo. Stem cell-derived adipocytes exposed to caffeine (1 mM) showed increased UCP1 protein abundance and cell metabolism with enhanced oxygen consumption and proton leak. These functional responses were associated with browning-like structural changes in mitochondrial and lipid droplet content. Caffeine also increased peroxisome proliferator-activated receptor gamma coactivator 1-alpha expression and mitochondrial biogenesis, together with a number of BAT selective and beige gene markers. In vivo, drinking coffee (but not water) stimulated the temperature of the supraclavicular region, which co-locates to the main region of BAT in adult humans, and is indicative of thermogenesis. Taken together, these results demonstrate that caffeine can promote BAT function at thermoneutrality and may have the potential to be used therapeutically in adult humans

Physiological, Biochemical, and Transcriptional Responses to Single and Combined Abiotic Stress in Stress-Tolerant and Stress-Sensitive Potato Genotypes

Potato production is often constrained by abiotic stresses such as drought and high temperatures which are often present in combination. In the present work, we aimed to identify key mechanisms and processes underlying single and combined abiotic stress tolerance by comparative analysis of tolerant and susceptible cultivars. Physiological data indicated that the cultivars Desiree and Unica were stress tolerant while Agria and Russett Burbank were stress susceptible. Abiotic stress caused a greater reduction of photosynthetic carbon assimilation in the susceptible cultivars which was associated with a lower leaf transpiration rate. Oxidative stress, as estimated by the accumulation of malondialdehyde was not induced by stress treatments in any of the genotypes with the exception of drought stress in Russett Burbank. Stress treatment resulted in increases in ascorbate peroxidase activity in all cultivars except Agria which increased catalase activity in response to stress. Transcript profiling highlighted a decrease in the abundance of transcripts encoding proteins associated with PSII light harvesting complex in stress tolerant cultivars. Furthermore, stress tolerant cultivars accumulated fewer transcripts encoding a type-1 metacaspase implicated in programmed cell death. Stress tolerant cultivars exhibited stronger expression of genes associated with plant growth and development, hormone metabolism and primary and secondary metabolism than stress susceptible cultivars. Metabolite profiling revealed accumulation of proline in all genotypes following drought stress that was partially suppressed in combined heat and drought. On the contrary, the sugar alcohols inositol and mannitol were strongly accumulated under heat and combined heat and drought stress while galactinol was most strongly accumulated under drought. Combined heat and drought also resulted in the accumulation of Valine, isoleucine, and lysine in all genotypes. These data indicate that single and multiple abiotic stress tolerance in potato is associated with a maintenance of CO2 assimilation and protection of PSII by a reduction of light harvesting capacity. The data further suggests that stress tolerant cultivars suppress cell death and maintain growth and development via fine tuning of hormone signaling, and primary and secondary metabolism. This study highlights potential targets for the development of stress tolerant potato cultivars

A member of the TERMINAL FLOWER1/CENTRORADIALIS gene family controls sprout growth in potato tubers

Potato tuber bud dormancy break followed by premature sprouting is a major commercial problem which results in quality losses and decreased tuber marketability. An approach to controlling premature tuber sprouting is to develop potato cultivars with a longer dormancy period and/or reduced rate of sprout growth. Our recent studies using a potato diploid population have identified several quantitative trait loci (QTLs) that are associated with tuber sprout growth. In the current study, we aim to characterize a candidate gene associated with one of the largest effect QTLs for rapid tuber sprout growth on potato chromosome 3. Underlying this QTL is a gene encoding a TERMINAL FLOWER 1/CENTRORADIALIS homologue (PGSC0003DMG400014322). Here, we use a transgenic approach to manipulate the expression level of the CEN family member in a potato tetraploid genotype (cv. Désirée). We demonstrate a clear effect of manipulation of StCEN expression, with decreased expression levels associated with an increased rate of sprout growth, and overexpressing lines showing a lower rate of sprout growth than controls. Associated with different levels of StCEN expression were different levels of abscisic acid and cytokinins, implying a role in controlling the levels of plant growth regulators in the apical meristem

Expression profiling of potato germplasm differentiated in quality traits leads to the identification of candidate flavour and texture genes

Quality traits such as flavour and texture are assuming a greater importance in crop breeding programmes. This study takes advantage of potato germplasm differentiated in tuber flavour and texture traits. A recently developed 44 000-element potato microarray was used to identify tuber gene expression profiles that correspond to differences in tuber flavour and texture as well as carotenoid content and dormancy characteristics. Gene expression was compared in two Solanum tuberosum group Phureja cultivars and two S. tuberosum group Tuberosum cultivars; 309 genes were significantly and consistently up-regulated in Phureja, whereas 555 genes were down-regulated. Approximately 46% of the genes in these lists can be identified from their annotation and amongst these are candidates that may underpin the Phureja/Tuberosum trait differences. For example, a clear difference in the cooked tuber volatile profile is the higher level of the sesquiterpene α-copaene in Phureja compared with Tuberosum. A sesquiterpene synthase gene was identified as being more highly expressed in Phureja tubers and its corresponding full-length cDNA was demonstrated to encode α-copaene synthase. Other potential ‘flavour genes’, identified from their differential expression profiles, include those encoding branched-chain amino acid aminotransferase and a ribonuclease suggesting a mechanism for 5′-ribonucleotide formation in potato tubers on cooking. Major differences in the expression levels of genes involved in cell wall biosynthesis (and potentially texture) were also identified, including genes encoding pectin acetylesterase, xyloglucan endotransglycosylase and pectin methylesterase. Other gene expression differences that may impact tuber carotenoid content and tuber life-cycle phenotypes are discussed

Fifteen new risk loci for coronary artery disease highlight arterial-wall-specific mechanisms

Coronary artery disease (CAD) is a leading cause of morbidity and mortality worldwide. Although 58 genomic regions have been associated with CAD thus far, most of the heritability is unexplained, indicating that additional susceptibility loci await identification. An efficient discovery strategy may be larger-scale evaluation of promising associations suggested by genome-wide association studies (GWAS). Hence, we genotyped 56,309 participants using a targeted gene array derived from earlier GWAS results and performed meta-analysis of results with 194,427 participants previously genotyped, totaling 88,192 CAD cases and 162,544 controls. We identified 25 new SNP-CAD associations (P < 5 × 10(-8), in fixed-effects meta-analysis) from 15 genomic regions, including SNPs in or near genes involved in cellular adhesion, leukocyte migration and atherosclerosis (PECAM1, rs1867624), coagulation and inflammation (PROCR, rs867186 (p.Ser219Gly)) and vascular smooth muscle cell differentiation (LMOD1, rs2820315). Correlation of these regions with cell-type-specific gene expression and plasma protein levels sheds light on potential disease mechanisms