Lipid droplet-organelle interactions: emerging roles in lipid metabolism.

Cellular homeostasis depends on the precisely coordinated use of lipids as fuels for energy production, building blocks for membrane biogenesis or chemical signals for intra-cellular and inter-cellular communication. Lipid droplets (LDs) are universally conserved dynamic organelles that can store and mobilize fatty acids and other lipid species for their multiple cellular roles. Increasing evidence suggests that contact zones between LDs and other organelles play important roles in the trafficking of lipids and in the regulation of lipid metabolism. Here we review recent advances regarding the nature and functional relevance of interactions between LDs and other organelles-particularly the endoplasmic reticulum (ER), LDs, mitochondria and vacuoles-that highlight their importance for lipid metabolism.This work was supported by an MRC Senior Non-Clinical Fellowship (to SS) and a Wellcome Trust Senior Clinical Fellowship (to DBS).This is the accepted manuscript. The final version is available at http://www.sciencedirect.com/science/article/pii/S0955067415000575

Spatial distribution of lipid droplets during starvation: Implications for lipophagy.

Survival during starvation depends largely on metabolic energy, which is stored in the form of neutral lipids in specialized organelles known as lipid droplets. The precursors for the synthesis of neutral lipids are also used for membrane biogenesis, which is required for cell growth and proliferation. Therefore cells must possess mechanisms to preferentially channel lipid precursors toward either membrane synthesis or lipid droplet storage, in response to nutrient status. How this partitioning is spatially regulated within the endoplasmic reticulum (ER) where lipid droplets co-localize, remains poorly understood. We have recently shown that at the onset of starvation lipid droplets concentrate at a perinuclear ER subdomain flanking the nucleus-vacuole junction (NVJ) and that this is crucial for maintaining proper nuclear shape and ER membrane organization. Here we show that disruption of the NVJ does not block the translocation and internalization of lipid droplets into the vacuole for their degradation, which takes place at later stages of starvation. We propose that alternative pathways of lipid droplet translocation from the ER to the vacuole may exist to enable stationary phase-induced lipophagy.This work was supported by the Medical Research Council (G0701446) and the Wellcome Trust (108042/Z/15/Z)

An Erg11 lanosterol 14-α-demethylase-Arv1 complex is required for Candida albicans virulence.

Azole resistant fungal infections remain a health problem for the immune compromised. Current therapies are limited due to rises in new resistance mechanisms. Therefore, it is important to identify new drug targets for drug discovery and novel therapeutics. Arv1 (are1 are2 required for viability 1) function is highly conserved between multiple pathogenic fungal species. Candida albicans (C. albicans) cells lacking CaArv1 are azole hypersusceptible and lack virulence. Saccharomyces cerevisiae (S. cerevisiae) Scarv1 cells are also azole hypersusceptible, a phenotype reversed by expression of CaArv1, indicating conservation in the molecular mechanism for azole susceptibility. To define the relationship between Arv1 function and azole susceptibility, we undertook a structure/function analysis of ScArv1. We identified several conserved amino acids within the ScArv1 homology domain (ScAhd) required for maintaining normal azole susceptibility. Erg11 lanosterol 14-α-demethylase is the rate-limiting enzyme in sterol biosynthesis and is the direct target of azole antifungals, so we used our ScArv1 mutants in order to explore the relationship between ScArv1 and ScErg11. Specific ScArv1 mutants ectopically expressed from a low copy plasmid were unable to restore normal azole susceptibility to Scarv1 cells and had reduced Erg11 protein levels. Erg11 protein stability depended on its ability to form a heterodimeric complex with Arv1. Complex formation was required for maintaining normal azole susceptibility. Scarv1 cells expressing orthologous CaArv1 mutants also had reduced CaErg11 levels, were unable to form a CaArv1-CaErg11 complex, and were azole hypersusceptible. Scarv1 cells expressing CaArv1 mutants unable to interact with CaErg11 could not sustain proper levels of the azole resistant CaErg11Y132F F145L protein. Caarv1/Caarv1 cells expressing CaArv1 mutants unable to interact with CaErg11 were found to lack virulence using a disseminated candidiasis mouse model. Expressing CaErg11Y132F F145L did not reverse the lack of virulence. We hypothesize that the role of Arv1 in Erg11-dependent azole resistance is to stabilize Erg11 protein level. Arv1 inhibition may represent an avenue for treating azole resistance

Lipid partitioning at the nuclear envelope controls membrane biogenesis.

Partitioning of lipid precursors between membranes and storage is crucial for cell growth, and its disruption underlies pathologies such as cancer, obesity, and type 2 diabetes. However, the mechanisms and signals that regulate this process are largely unknown. In yeast, lipid precursors are mainly used for phospholipid synthesis in nutrient-rich conditions in order to sustain rapid proliferation but are redirected to triacylglycerol (TAG) stored in lipid droplets during starvation. Here we investigate how cells reprogram lipid metabolism in the endoplasmic reticulum. We show that the conserved phosphatidate (PA) phosphatase Pah1, which generates diacylglycerol from PA, targets a nuclear membrane subdomain that is in contact with growing lipid droplets and mediates TAG synthesis. We find that cytosol acidification activates the master regulator of Pah1, the Nem1-Spo7 complex, thus linking Pah1 activity to cellular metabolic status. In the absence of TAG storage capacity, Pah1 still binds the nuclear membrane, but lipid precursors are redirected toward phospholipids, resulting in nuclear deformation and a proliferation of endoplasmic reticulum membrane. We propose that, in response to growth signals, activation of Pah1 at the nuclear envelope acts as a switch to control the balance between membrane biogenesis and lipid storage.This work was supported by grants from the Medical Research Council (G0701446) to S.S; a Wellcome Trust Strategic Award (100140) and equipment grant (093026) to the Cambridge Institute for Medical Research; the National Institutes of Health (GM050679) to G.M.C.; a ALW Open Program (822.02.014), DFG-NWO cooperation (DN82-303), SNSF Sinergia (CRSII3_154421) and ZonMW VICI (016.130.606) grants to F.R; and a PhD fellowship from the Fundação para a Ciência e a Tecnologia (FCT) to S.A.This is the final version of the article. It first appeared from the American Society for Cell Biology via http://dx.doi.org/10.1091/mbc.E15-03-017

Schoolbag weight carriage in Portuguese children and adolescents: a cross-sectional study comparing possible influencing factors

Background Schoolbags and the consequences of carrying them, particularly those associated with overload, are often studied as a health concern. Modifications in gait and posture were reported when children carried loads that corresponded to more than 10% of their body weight (BW). The aims of this study were to verify the load that is carried by Portuguese students and how it is influenced by factors such as school grade, school schedule, lunch site, physical education, sex and body mass index (BMI). Acquiring a more specific knowledge of the Portuguese context and understanding the influence of these factors may allow us to generate proposals to control them in ways that benefit students.MethodsThe load carried by students in the 5th grade (10.60.4years) and 9th grade (14.70.6years) were weighed with a luggage scale on all days of the week, resulting in 680 evaluations. Data related to the school day were also collected, such as the student's lunch site, how he or she got to school and his or her school schedule for that day. Individual height and weight were also assessed. Results The 5th grade students carried greater loads than the 9th grade students, resulting in a substantial difference relative to their BW. The school loads of the 5th grade students were mostly greater than 10% of their BWs. Girls tended to carry heavier loads than boys, and overweight students also tended to carry heavier loads. Students who could eat lunch at home carried less weight, and on physical education days, the total load carried increased, but the backpacks of the 5th grade students were lighter.Conclusions The results of the current study describe excessive schoolbag weight among Portuguese students and expound on some of the factors that influence it, which can help researchers and professionals design a solution to decrease children's schoolbag loads.Funds through FCT - Portuguese Foundation for Science and Technology (UID/DTP/04045/2019) - and the European Fund for regional development (FEDER) allocated by European Union through the COMPETE 2020 Programme (POCI-01-0145-FEDER-006969), and through the Project NanoSTIMA: Macro-to-Nano Human Sensing, Towards Integrated Multimodal Health Monitoring and Analytics (NORTE-01-0145-FEDER-000016), co-financed by Fundo Europeu de Desenvolvimento Regional (FEDER) -NORTE 2020

MOTIVOS DE ADESÃO, MANUTENÇÃO E RESULTADOS ALCANÇADOS ATRAVÉS DA PRÁTICA DE ATIVIDADES FÍSICAS EM ACADEMIAS

Este estudo teve como objetivo identificar e analisar os motivos que levam jovens e adultos a aderirem e manterem-se em programas regulares de atividades físicas oferecidos em duas academias. Para a coleta de dados desta pesquisa utilizou-se um questionário específico contendo perguntas fechadas, o qual foi aplicado a 120 sujeitos de ambos os sexos, faixa etária entre 15 e 32 anos, abrangendo adolescentes e adultos. Analisando os resultados, concluí-se que a maioria dos sujeitos analisados, possui tempo de prática considerado bom e a freqüência semanal adequada. Os motivos que levaram os grupos analisados a aderir e a manter a prática de atividades físicas em academias referem-se à estética corporal, aptidão física e outros, considerando que a pratica de atividades físicas contribuem para a melhora da qualidade de vida e da saúde do homem desde a infância, adolescência, na vida adulta e na maior idade

Forensic neuropsychological assessment: a review of its scope

Background Issues related to the field of mental health and justice require a multifactorial understanding of the possible causes of such issues. Objective To conduct an integrative literature review of controlled studies describing forensic neuropsychological assessment. Methods The articles were compiled and analyzed in two phases: 1) first, we retrieved all papers in PubMed by the keywords “Forensic Neuropsychology” and generated a growth curve for the subject and a cluster-based thematic distribution of publications. 2) We then conducted a curated analysis of all relevant papers indexed in Medline, PubMed and ISI, between 2000 and 2012. Results The evolution of the field during the last 15 years reveals an unstable growth pattern and three main thematic clusters. In terms of our curated analysis, a total of 390 articles were pre-selected, resulting in the selection of 44 fully-relevant studies, which comprise four main categories: cognitive damage in forensic psychiatric patients; imitation of cognitive damage; civil capacity, penal liability and violence risk; and validation of neuropsychological assessment tools. Discussion Two aspects appeared as the most relevant in this study: growth in the use of neuropsychological assessment as a diagnostic tool in the forensic context; and the necessity to enhance conformity in assessments

Impact of occlusion duration on the success rate and outcomes of percutaneous coronary intervention in chronic total occlusions

ABSTRACTBackgroundInitial studies have shown that old occlusions or those with indeterminate occlusion duration have been associated with percutaneous coronary intervention (PCI) failure and a worse prognosis. This study aimed to determine the impact of occlusion duration on the success and outcomes of contemporary PCI on chronic total occlusion (CTO).MethodsThe authors analyzed a retrospective cohort of consecutive patients submitted to PCI in CTO, who were compared according to the confirmed occlusion duration (COD) < 12 months, ≥ 12 months, or indeterminate occlusion duration (IOD).ResultsA total of 168 patients were treated, 122 (72.6%) with COD (80 < 12 months, 42 ≥ 12 months) and 46 (24.7%) with an IOD. Lesion extension was 17.0 ± 13.6mm, in 2.90 ± 0.58mm vessels, and the anterograde approach was used in 98.8% of cases. Angiographic success was attained in 79.2% of patients (80.0% vs. 73.8% vs. 82.6%; p = 0.73). The main cause of failure was the inability to cross the lesion with the guidewire (68.6%). Occlusion duration had no impact on in-hospital events (4.8% vs. 7.1% vs. 6.0%; p = 0.73), which were almost entirely explained by periprocedural myocardial infarction, or on late outcomes (18.8% vs. 7.1% vs. 15.3%; p = 0.23). At the multivariate analysis, lesion length ≥ 20mm (odds ratio - OR = 7.27; 95% confidence interval - 95% IC 1.94-29.1; p = 0.003), calcification (OR = 4.72; 95% CI 1.19-19.1; p = 0.02), and tortuosity of the occluded segment (OR = 15.98; 95% CI 2.18-144.7; p = 0.007) were predictors of failure.ConclusionsOcclusion duration was not associated with increased failure rate of the procedure or worse PCI outcomes in CTO