Optimization of de novo 10-hydroxygeraniol production in Saccharomyces cerevisiae

Monoterpenes and their indole alkaloid derivates are plant secondary metabolites valuable to the pharmaceutical and industrial sectors for their bioactivities in humans, cats and insects. The biosynthetic pathway to strictosidine, the central monoterpene indole alkaloid scaffold, proceeds via an industrially relevant intermediate – nepetalactol. The oxidized counterpart of this monoterpene, nepetalactone, is known for its euphoric influence on cats but is also a potent insect repellent. Unfortunately, low in planta levels of these metabolites have resulted in high production costs and environmentally unfeasible cultivation strategies for an industrial product with mass market appeal. Engineering a high-producing nepetalactol or strictosidine yeast would benefit several commercial sectors, however recent studies attempting de novo strictosidine in S. cerevisiae only achieved trace titers (0.5 mg/L). Activity of geraniol-10-hydroxylase, a cytochrome P450, was an identified bottleneck in nepetalactol/strictosidine production. This thesis presents the application of a combination of strategies aimed at overcoming the geraniol-10-hydroxylase bottleneck and improving titres of 10-hydroxygeraniol. Geraniol-10-hydroxylase and cytochrome P450 reductase variants were explored to increase enzyme efficiency, wherein several reductases were shown to beneficially influence hydroxylation activity. We further demonstrated that a cytochrome P450-reductase fusion increased carbon flux towards 10-hydroxygeraniol by 1.7-fold. Accumulation of a heterologous metabolite, putatively identified as isopulegol, was observed, implying inefficient channeling of carbon into the heterologous pathway. Deletion of two native reductases, oye2 and oye3, resulted in a 4.6-fold increase in carbon channeled to 10-hydroxygeraniol, resulting in a final accumulation of 144 mg/L of 10-hydroxygeraniol. These pathway optimization steps will aid in the development of high yielding monoterpene S. cerevisiae strains

Perspectives et place des pères dans les services de protection de l’enfance et de la jeunesse

Parmi les nouveaux courants de recherche et les pratiques émergentes en travail social auprès des familles, se retrouve la question de la place des pères dans les familles et dans les interventions. Par ailleurs, cette reconceptualisation de la paternité, tant sur le plan théorique que pratique, s’accompagne de tensions entre différentes idéologies. Notamment, selon que l’on adhère à une définition plutôt conservatrice, féministe ou liée aux droits des pères, ces derniers sont appelés à jouer des rôles forts différents auprès des mères et des enfants. La façon dont les travailleurs sociaux se situent face à ces tendances induira différentes façons d’accompagner les familles et les pères. Il s’avère donc pertinent d’étudier les courants de pensée présents dans ce champ de façon, à savoir la place qu’ils réservent aux besoins des pères. Ultimement, cette réflexion pourrait susciter le développement de nouvelles approches d’intervention en travail social auprès des pères. Cette question est abordée dans cet article sous l’angle des services de protection de l’enfance et de la jeunesse

Research Priorities in Pediatric Palliative Care: A Delphi Study

Pediatric palliative care is increasingly recognized to be a specialized type of care requiring specific skills and knowledge, yet, as found in several countries, there is little available research evidence on which to base care. Objectives: The goal of the project was to achieve consensus among palliative care practitioners and researchers regarding the identification of pertinents lines of research. Method: A Delphi technique was used with an interdisciplinary panel (n=14–16) of researchers and frontline clinicians in pediatric palliative care in Canada. Results: Four priority research questions were identified: What matters most for patients and parents receiving pediatric palliative services? What are the bereavement needs of families in pediatric palliative care? What are the best practice standards in pain and symptom management? What are effective strategies to alleviate suffering at the end of life? Conclusions: These identified priorities will provide guidance and direction for research efforts in Canada, and may prove useful in providing optimal care to patients and families in pediatric palliative care

FOXD3 Regulates VISTA Expression in Melanoma.

Immune checkpoint inhibitors have improved patient survival in melanoma, but the innate resistance of many patients necessitates the investigation of alternative immune targets. Many immune checkpoint proteins lack proper characterization, including V-domain Ig suppressor of T cell activation (VISTA). VISTA expression on immune cells can suppress T cell activity; however, few studies have investigated its expression and regulation in cancer cells. In this study, we observe that VISTA is expressed in melanoma patient samples and cell lines. Tumor cell-specific expression of VISTA promotes tumor onset in vivo, associated with increased intratumoral T regulatory cells, and enhanced PDL-1 expression on tumor-infiltrating macrophages. VISTA transcript levels are regulated by the stemness factor Forkhead box D3 (FOXD3). BRAF inhibition upregulates FOXD3 and reduces VISTA expression. Overall, this study demonstrates melanoma cell expression of VISTA and its regulation by FOXD3, contributing to the rationale for therapeutic strategies that combine targeted inhibitors with immune checkpoint blockade

Multicenter clinical evaluation of Etest meropenem-vaborbactam (bioMérieux) for susceptibility testing of Enterobacterales (Enterobacteriaceae) and Pseudomonas aeruginosa

Meropenem-vaborbactam (MEV) is a novel carbapenem-beta-lactamase inhibitor combination antibiotic approved by the U.S. Food and Drug Administration (FDA) for treatment of complicated urinary tract infections, including pyelonephritis, in adults. In this study, we evaluated the performance of Etest MEV (bioMérieux, Marcy l\u27Etoile, France) compared to that of broth microdilution for 62

Current and Emerging Uses of Statins in Clinical Therapeutics: A Review

Statins, a class of cholesterol-lowering medications that inhibit 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase, are commonly administered to treat atherosclerotic cardiovascular disease. Statin use may expand considerably given its potential for treating an array of cholesterol-independent diseases. However, the lack of conclusive evidence supporting these emerging therapeutic uses of statins brings to the fore a number of unanswered questions including uncertainties regarding patient-to-patient variability in response to statins, the most appropriate statin to be used for the desired effect, and the efficacy of statins in treating cholesterol-independent diseases. In this review, the adverse effects, costs, and drug–drug and drug–food interactions associated with statin use are presented. Furthermore, we discuss the pleiotropic effects associated with statins with regard to the onset and progression of autoimmune and inflammatory diseases, cancer, neurodegenerative disorders, strokes, bacterial infections, and human immunodeficiency virus. Understanding these issues will improve the prognosis of patients who are administered statins and potentially expand our ability to treat a wide variety of diseases

SUMO is a pervasive regulator of meiosis

Protein modification by SUMO helps orchestrate the elaborate events of meiosis to faithfully produce haploid gametes. To date, only a handful of meiotic SUMO targets have been identified. Here, we delineate a multidimensional SUMO-modified meiotic proteome in budding yeast, identifying 2747 conjugation sites in 775 targets, and defining their relative levels and dynamics. Modified sites cluster in disordered regions and only a minority match consensus motifs. Target identities and modification dynamics imply that SUMOylation regulates all levels of chromosome organization and each step of meiotic prophase I. Execution-point analysis confirms these inferences, revealing functions for SUMO in S-phase, the initiation of recombination, chromosome synapsis and crossing over. K15-linked SUMO chains become prominent as chromosomes synapse and recombine, consistent with roles in these processes. SUMO also modifies ubiquitin, forming hybrid oligomers with potential to modulate ubiquitin signaling. We conclude that SUMO plays diverse and unanticipated roles in regulating meiotic chromosome metabolism

Affect labeling: a promising new neuroscience-based approach to treating combat-related PTSD in veterans

IntroductionA significant portion of individuals exposed to combat-related trauma will develop posttraumatic stress disorder (PTSD), a severe, debilitating disorder with adverse impacts on both mental and physical functioning. Current treatments are effective for many individuals, however, there is a need for new treatment approaches to improve outcomes in PTSD and address the many existing barriers to seeking or completing treatment.MethodsIn this open trial pilot study, we tested a novel, brief, computer-based intervention for PTSD utilizing “affect labeling” that was inspired by recent advances in neuroscience with U.S. veterans.ResultsAs expected, pre-intervention clinical and fMRI neuroimaging data indicated that U.S. veterans with combat-related PTSD (N = 20) had significantly higher PTSD symptoms, depression symptoms, and amygdala reactivity to trauma cues than trauma-exposed healthy control veterans (N = 20). Veterans with PTSD who completed the affect labeling intervention (N = 13) evidenced reduced PTSD symptoms and these reductions were correlated with reductions in amygdala reactivity.DiscussionResults from this initial proof-of-concept study are intriguing and suggest that affect labeling training offers significant potential as a novel, cost-effective, computer-based intervention for PTSD. Implications and next steps for further developing affect labeling interventions for PTSD are discussed.Clinical Trial Registrationhttps://clinicaltrials.gov/, identifier NCT05924399

Methicillin-Resistant Staphylococcus aureus ST9 in Pigs in Thailand

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) is an important nosocomial and community-associated pathogen. Recently, livestock-associated MRSA (LA-MRSA) has emerged and disseminated in Europe and North America and now constitutes a considerable zoonotic burden in humans with risk factors of pig exposure, whereas the extent of the livestock reservoir is relatively unknown on other continents. METHODOLOGY/PRINCIPAL FINDINGS: From March through April 2011, MRSA was identified in pigs from 3 out of 30 production holdings in Chang Mai Province, Thailand. Representative isolates were subjected to molecular characterization and antimicrobial susceptibility testing; all isolates had genotypic and phenotypic characteristics of LA-MRSA previously characterized in the region: they belonged to ST9, lacked the lukF-lukS genes encoding Panton-Valentine leukocidin, and were resistant to multiple non-β-lactam antimicrobials. However, unlike other Asian LA-MRSA-ST9 variants, they were spa type t337 and harbored a different staphylococcal cassette chromosome mec IX. CONCLUSIONS/SIGNIFICANCE: A novel MRSA-ST9 lineage has been established in the pig population of Thailand, which differs substantially from LA-MRSA lineages found in other areas of the continent. The emergence of novel LA-MRSA lineages in the animal agriculture setting is worrisome and poses a serious threat to global public health

2016 Research & Innovation Day Program

A one day showcase of applied research, social innovation, scholarship projects and activities.https://first.fanshawec.ca/cri_cripublications/1003/thumbnail.jp