Loop-mediated isothermal amplification: Development, validation and application of simple and rapid assays for quantitative detection of species of Arcobacteraceae family- and species-specific Aliarcobacter faecis and Aliarcobacter lanthieri.

AIM: The family Arcobacteraceae formerly genus Arcobacter has recently been reclassified into six genera. Among nine species of the genus Aliarcobacter, Aliarcobacter faecis and Aliarcobacter lanthieri have been identified as emerging pathogens potentially cause health risks to humans and animals. This study was designed to develop/optimize, validate and apply Arcobacteraceae family- and two species-specific (A. faecis and A. lanthieri) loop-mediated isothermal amplification (LAMP) assays to rapidly detect and quantify total number of cells in various environmental niches. METHODS AND RESULTS: Three sets of LAMP primers were designed from conserved and variable regions of 16S rRNA (family-specific) and gyrB (species-specific) genes. Optimized Arcobacteraceae family-specific LAMP assay correctly amplified and detected 24 species, whereas species-specific LAMP assays detected A. faecis and A. lanthieri reference strains as well as 91 pure and mixed culture isolates recovered from aquatic and faecal sources. The specificity of LAMP amplification of A. faecis and A. lanthieri was further confirmed by restriction fragment length polymorphism analysis. Assay sensitivities were tested using variable DNA concentrations extracted from simulated target species cells in an autoclaved agricultural water sample by achieving a minimum detection limit of 10 cells mL-1 (10 fg). Direct DNA-based quantitative detection, from agricultural surface water, identified A. faecis (17%) and A. lanthieri (1%) at a low frequency compared to family-level (93%) with the concentration ranging from 2·1 × 101 to 2·2 × 105 cells 100 mL-1 . CONCLUSIONS: Overall, these three DNA-based rapid and cost-effective novel LAMP assays are sensitive and can be completed in less than 40 min. They have potential for on-site quantitative detection of species of family Arcobacteraceae, A. faecis and A. lanthieri in food, environmental and clinical matrices. SIGNIFICANCE AND IMPACT OF THE STUDY: The newly developed LAMP assays are specific, sensitive, accurate with higher reproducibility that have potential to facilitate in a less equipped lab setting and can help in early quantitative detection and rate of prevalence in environmental niches. The assays can be adopted in the diagnostic labs and epidemiological studies.

Managing asthma in adolescents and adults 2020 asthma guideline update From the National Asthma Education and Prevention Program

Asthma is a major public health problem worldwide and is associated with excess morbidity, mortality, and economic costs associated with lost productivity. The National Asthma Education and Prevention Program has released the 2020 Asthma Guideline Update with updated evidence-based recommendations for treatment of patients with asthma. To report updated recommendations for 6 topics for clinical management of adolescents and adults with asthma: (1) intermittent inhaled corticosteroids (ICSs); (2) add-on long-acting muscarinic antagonists; (3) fractional exhaled nitric oxide; (4) indoor allergen mitigation; (5) immunotherapy; and (6) bronchial thermoplasty. The National Heart, Lung, and Blood Advisory Council chose 6 topics to update the 2007 asthma guidelines based on results from a 2014 needs assessment. The Agency for Healthcare Research and Quality conducted systematic reviews of these 6 topics based on literature searches up to March-April 2017. Reviews were updated through October 2018 and used by an expert panel (n = 19) that included asthma content experts, primary care clinicians, dissemination and implementation experts, and health policy experts to develop 19 new recommendations using the GRADE method. The 17 recommendations for individuals aged 12 years or older are reported in this Special Communication. From 20 572 identified references, 475 were included in the 6 systematic reviews to form the evidence basis for these recommendations. Compared with the 2007 guideline, there was no recommended change in step 1 (intermittent asthma) therapy (as-needed short-acting ß2-agonists [SABAs] for rescue therapy). In step 2 (mild persistent asthma), either daily low-dose ICS plus as-needed SABA therapy or as-needed concomitant ICS and SABA therapy are recommended. Formoterol in combination with an ICS in a single inhaler (single maintenance and reliever therapy) is recommended as the preferred therapy for moderate persistent asthma in step 3 (low-dose ICS-formoterol therapy) and step 4 (medium-dose ICS-formoterol therapy) for both daily and as-needed therapy. A short-term increase in the ICS dose alone for worsening of asthma symptoms is not recommended. Add-on long-acting muscarinic antagonists are recommended in individuals whose asthma is not controlled by ICS-formoterol therapy for step 5 (moderate-severe persistent asthma). Fractional exhaled nitric oxide testing is recommended to assist in diagnosis and monitoring of symptoms, but not alone to diagnose or monitor asthma. Allergen mitigation is recommended only in individuals with exposure and relevant sensitivity or symptoms. When used, allergen mitigation should be allergen specific and include multiple allergen-specific mitigation strategies. Subcutaneous immunotherapy is recommended as an adjunct to standard pharmacotherapy for individuals with symptoms and sensitization to specific allergens. Sublingual immunotherapy is not recommended specifically for asthma. Bronchial thermoplasty is not recommended as part of standard care; if used, it should be part of an ongoing research effort. Asthma is a common disease with substantial human and economic costs globally. Although there is no cure or established means of prevention, effective treatment is available. Use of the recommendations in the 2020 Asthma Guideline Update should improve the health of individuals with asthma.

Mapping responsible conduct in the uncharted field of research-creation: A scoping review.

This scoping review addresses the issues of responsible conduct of research (RCR) that can arise in the practice of research-creation (RC), an emergent, interdisciplinary, and heterogeneous field at the interface of academic research and creative activities. Little is yet known about the nature and scope of RCR issues in RC, so our study examined three questions: (1) What are the specific issues in RC in relation to RCR? (2) How does the specificity of RC influence the understanding and practice of RCR? (3) What recommendations could help address the issues highlighted in the literature? To answer these questions, we conducted a scoping review of the academic literature (n = 181 texts) dealing with RCR in RC. We found that researcher-creators faced some very different RCR challenges in comparison with their colleagues in the rest of academia. Addressing these issues is important for both the RCR and RC communities in order to ensure that the rapid development of this field occurs in line with the norms of RCR which, nonetheless, should be adapted to respect the particularities of RC and allow its contributions to the academic world.

Outcomes of an early childhood obesity prevention program in a low-income community: a pilot, randomized trial.

BACKGROUND: Obesity is a major and worldwide health problem in children. OBJECTIVES: The Early Childhood Obesity Prevention Program is a multi-component, randomized, controlled trial of a pilot community-focused obesity prevention program for mother/newborn dyads. METHODS: Underserved, mother/newborn dyads were recruited to receive a standard home visitation program (Nurturing Families Network, NFN) or an enhanced program (NFN+) that incorporated behavioural change strategies (e.g., goal-setting, problem-solving) and focused on six obesity-associated behaviours (breastfeeding, juice/sugar-sweetened beverages, solids, infant sleep, TV/screen time and soothability) with linkages to community resources. Weight-for-length (WFL) z-score and maternal diet were secondary outcomes. RESULTS: Fifty-seven dyads were recruited and 47 fully eligible dyads were enrolled (NFN = 21, NFN+ = 26). Forty-one (87.2%) were assessed at 6 months and 34 (72.3%) at 12 months. Retention at 12 months was higher for NFN+ dyads (84.6% vs. 56.1%, p = 0.04). NFN+ mothers were more likely to continue breastfeeding at 6 and 12 months vs. NFN mothers (p = 0.03 and 0.003, respectively), and at 12 months, NFN+ infants had fewer nocturnal awakenings (p = 0.04). There were no differences in other primary outcome measures or in WFL z-score at 6 or 12 months. CONCLUSIONS: A multi-component behavioural intervention increased breastfeeding duration and decreased nocturnal awakenings in infants of low-income families.

Enhanced Single-tube Multiplex PCR Assay for Detection and Identification of Six Arcobacter Species.

AIM: A single-tube multiplex PCR (mPCR) assay was developed for rapid, sensitive and simultaneous detection and identification of six Arcobacter species including two new species, A. lanthieri and A. faecis, along with A. butzleri, A. cibarius, A. cryaerophilus and A. skirrowii on the basis of differences in the lengths of their PCR products. Previously designed monoplex, mPCR and RFLP assays do not detect or differentiate A. faecis and A. lanthieri from other closely related known Arcobacter spp. METHODS AND RESULTS: Primer pairs for each target species (except A. skirrowii) and mPCR protocol were newly designed and optimized using variable regions of housekeeping including cpn60, gyrA, gyrB and rpoB genes. The accuracy and specificity of the mPCR assay was assessed using DNA templates from six targets and 11 other Arcobacter spp. as well as 50 other bacterial reference species and strains. Tests on the DNA templates of target Arcobacter spp. were appropriately identified, whereas all 61 other DNA templates from other bacterial species and strains were not amplified. Sensitivity and specificity of the mPCR assay was 10 pg µl-1 of DNA concentration per target species. The optimized assay was further evaluated, validated and compared with other mPCR assays by testing Arcobacter cultures isolated from various faecal and water sources. CONCLUSIONS: Study results confirm that the newly developed mPCR assay is rapid, accurate, reliable, simple, and valuable for the simultaneous detection and routine diagnosis of six human- and animal-associated Arcobacter spp. SIGNIFICANCE AND IMPACT OF THE STUDY: The new mPCR assay is useful not only for pure but also mixed cultures. Moreover, it has the ability to rapidly detect six species which enhances the value of this technology for aetiological and epidemiological studies.

Implementing a routine flow cytometry assay for nucleated red blood cell counts in cord blood units.

INTRODUCTION: As required by standards organizations, Héma-Québec Cord Blood Bank performs enumeration of nucleated red blood cells (NRBCs) in cord blood units (CBUs). This study presents the validation and implementation approaches developed to transfer the routine NRBC enumeration from the manual blood film method to a flow cytometric assay. METHODS: The flow cytometry method was adapted from Tsuji (Cytometry, 37, 1999, 291). This assay was validated to assess the specificity, detection limit, repeatability, and reproducibility of the method, including interoperator and interlaboratory testing. Finally, postimplementation follow-up and adjustments were performed for CBU over a 7-month period. RESULTS: Blood film and flow cytometry NRBC enumerations showed a strong correlation (n = 40; Pearson's r correlation = 0.90). Validation was successful as exemplified by the correlation in interlaboratory testing (n = 30; r = 0.98). During implementation, our routine laboratory analyses revealed that CBU with low NRBC content (≤2%), representing 26% of all CBU tested, resulted in 15% of repeated reading and/or staining and was the principal source of nonconformity. Small adjustments in the standard operating procedures (SOPs), including a fixed 200-event setting in the NRBC gate for the second reading of the replicates, have completely solved this issue. CONCLUSION: Flow cytometric NRBC enumerations, now implemented in Héma-Québec Public Cord Blood Bank, is an improvement in the efficiency of our operations by integrating the count for NRBC into our flow cytometry platform.

Controlled Distribution and Clustering of Silver in Ag-DLC Nanocomposite Coatings Using a Hybrid Plasma Approach.

Incorporation of selected metallic elements into diamond-like carbon (DLC) has emerged as an innovative approach to add unique functional properties to DLC coatings, thus opening up a range of new potential applications in fields as diverse as sensors, tribology, and biomaterials. However, deposition by plasma techniques of metal-containing DLC coatings with well-defined structural properties and metal distribution is currently hindered by the limited understanding of their growth mechanisms. We report here a silver-incorporated diamond-like carbon coating (Ag-DLC) prepared in a hybrid plasma reactor which allowed independent control of the metal content and the carbon film structure and morphology. Morphological and chemical analyses of Ag-DLC films were performed by atomic force microscopy, scanning electron microscopy, and X-ray photoelectron spectroscopy. The vertical distribution of silver from the surface toward the coating bulk was found to be highly inhomogeneous due to top surface segregation and clustering of silver nanoparticles. Two plasma parameters, the sputtered Ag flux and ion energy, were shown to influence the spatial distribution of silver particles. On the basis of these findings, a mechanism for Ag-DLC growth by plasma was proposed.

Antibacterial Coatings: Challenges, Perspectives, and Opportunities.

Antibacterial coatings are rapidly emerging as a primary component of the global mitigation strategy of bacterial pathogens. Thanks to recent concurrent advances in materials science and biotechnology methodologies, and a growing understanding of environmental microbiology, an extensive variety of options are now available to design surfaces with antibacterial properties. However, progress towards a more widespread use in clinical settings crucially depends on addressing the key outstanding issues. We review release-based antibacterial coatings and focus on the challenges and opportunities presented by the latest generation of these materials. In particular, we highlight recent approaches aimed at controlling the release of antibacterial agents, imparting multi-functionality, and enhancing long-term stability.

Red light violations by adult pedestrians and other safety-related behaviors at signalized crosswalks.

To study human factors linked to red light violations, and more generally to safety-related behaviors at signalized crosswalks, the present study combines the collection of observational data with questionnaires answered by 422 French adult pedestrians. Thirteen behavioral indicators were extracted (12 before and while crossing, and red light violation), and the roles of several demographical, contextual and mobility-associated variables were examined. The results of the stepwise logistic regression analyses carried out on each of the 12 behavioral indicators observed before and while crossing revealed that gender had no major impact, but age did, with more cautious behaviors as pedestrians were older. The three contextual variables (group size, parked vehicles, and traffic density), as four mobility-associated variables (driving and walking experiences, self-reported crossing difficulties and falls in the street) were also found to be important factors in safety-related crossing behaviors. A wider logistic regression analysis, made specifically on red light violations with all behavioral indicators observed before and while crossings and the several demographical, contextual and mobility-associated variables put together, showed that red light violations were mostly affected by current situational factors (group size, parked vehicles) and particularly associated with some behavioral patterns (looking toward the traffic, the ground, the light, running and crossing diagonally). The overall results encourage the development of safer pedestrian infrastructures and engineering countermeasures.

Blood donation clusters in Québec, Canada (2003-2008): spatial variations according to sex and age.

BACKGROUND AND OBJECTIVES: The detection of spatial clusters of blood donation rate is an important issue, especially for targeting spatial units with significantly low rates, where it could be possible to increase the numbers of donors. The objective of this study is to detect spatial clusters of high or low blood donation rate in Québec according to sex and age of the donors. MATERIALS AND METHODS: Blood donation data were obtained from Héma-Québec over a period of 5 years. We aggregated these data for each of 101 municipalités regionales de comté (i.e. counties) for men, women and four age groups. To detect spatial high/low donation rate areas, we used the Kulldorff's scan statistics. Kappa coefficient was used to assess discordance between clusters obtained for the different groups (18-29, 30-39, 40-49, 50-59, 60-69 years old). T-test analyses were conducted to identify significant associations between spatial clusters and socio-economic variables. RESULTS: The results indicate the presence of several geographical areas with high or low blood donation rates for each group. The size, the location and the socio-demographic profiles of low/high clusters vary according to sex and age categories. CONCLUSION: The Kulldorff's scan statistics are an efficient tool to assess the blood donation performance across a country or even a specific region over a period of several years. In terms of strategic planning and monitoring, it can be used as a fully operational tool to target areas with significantly low rates (for all donors or specific demographic groups) in future blood donation campaigns.

Genome-wide association study of body mass index in 23 000 individuals with and without asthma.

BACKGROUND: Both asthma and obesity are complex disorders that are influenced by environmental and genetic factors. Shared genetic factors between asthma and obesity have been proposed to partly explain epidemiological findings of co-morbidity between these conditions. OBJECTIVE: To identify genetic variants that are associated with body mass index (BMI) in asthmatic children and adults, and to evaluate if there are differences between the genetics of BMI in asthmatics and healthy individuals. METHODS: In total, 19 studies contributed with genome-wide analysis study (GWAS) data from more than 23 000 individuals with predominantly European descent, of whom 8165 are asthmatics. RESULTS: We report associations between several DENND1B variants (P = 2.2 × 10(-7) for rs4915551) on chromosome 1q31 and BMI from a meta-analysis of GWAS data using 2691 asthmatic children (screening data). The top DENND1B single nucleotide polymorphisms(SNPs) were next evaluated in seven independent replication data sets comprising 2014 asthmatics, and rs4915551 was nominally replicated (P < 0.05) in two of the seven studies and of borderline significance in one (P = 0.059). However, strong evidence of effect heterogeneity was observed and overall, the association between rs4915551 and BMI was not significant in the total replication data set, P = 0.71. Using a random effects model, BMI was overall estimated to increase by 0.30 kg/m(2) (P = 0.01 for combined screening and replication data sets, N = 4705) per additional G allele of this DENND1BSNP. FTO was confirmed as an important gene for adult and childhood BMI regardless of asthma status. CONCLUSIONS AND CLINICAL RELEVANCE: DENND1B was recently identified as an asthma susceptibility gene in a GWAS on children, and here, we find evidence that DENND1B variants may also be associated with BMI in asthmatic children. However, the association was overall not replicated in the independent data sets and the heterogeneous effect of DENND1B points to complex associations with the studied diseases that deserve further study.

Dynamic modelling of protein and oxidative metabolisms simulates the pathogenesis of Parkinson's disease.

Research into Parkinson's disease (PD) is difficult and time consuming. It is a complex condition that develops over many decades in the human brain. For such apparently intractable diseases, mathematical models can offer an additional means of investigation. As a contribution to this process, the authors have developed an ordinary differential equation model of the most important cellular processes that have been associated with PD. The model describes the following processes: (i) cellular generation and scavenging of reactive oxygen species; (ii) the possible damage and removal of the protein -synuclein and, (iii) feedback interactions between damaged α-synuclein and reactive oxygen species. Simulation results show that the Parkinsonian condition, with elevated oxidative stress and misfolded α-synuclein accumulation, can be induced in the model by known PD risk factors such as ageing, exposure to toxins and genetic defects. The significant outcome of the paper is the demonstration that it is possible to reproduce in silico the multi-factorial interactions that characterise the pathogenesis of PD. As such, the model provides a systematic explanation of the variability and heterogeneity of PD and provides the basis for computational studies of further facets of this complex multi-factorial condition. [Includes supplementary material].

Feedback motif for the pathogenesis of Parkinson's disease.

Previous article on the integrative modelling of Parkinson's disease (PD) described a mathematical model with properties suggesting that PD pathogenesis is associated with a feedback-induced biochemical bistability. In this article, the authors show that the dynamics of the mathematical model can be extracted and distilled into an equivalent two-state feedback motif whose stability properties are controlled by multi-factorial combinations of risk factors and genetic mutations associated with PD. Based on this finding, the authors propose a principle for PD pathogenesis in the form of the switch-like transition of a bistable feedback process from 'healthy' homeostatic levels of reactive oxygen species and the protein α-synuclein, to an alternative 'disease' state in which concentrations of both molecules are stable at the damagingly high-levels associated with PD. The bistability is analysed using the rate curves and steady-state response characteristics of the feedback motif. In particular, the authors show how a bifurcation in the feedback motif marks the pathogenic moment at which the 'healthy' state is lost and the 'disease' state is initiated. Further analysis shows how known risks (such as: age, toxins and genetic predisposition) modify the stability characteristics of the feedback motif in a way that is compatible with known features of PD, and which explain properties such as: multi-factorial causality, variability in susceptibility and severity, multi-timescale progression and the special cases of familial Parkinson's and Parkinsonian symptoms induced purely by toxic stress.

Parametric sensitivity analysis and reduction of a detailed nutritional model of plant cell cultures.

A kinetic model of plant nutrition described by Cloutier et al. (Cloutier et al., 2008. Biotechnol Bioeng 99:189-200) is progressively simplified so as to obtain a predictive model that describes the evolution of the biomass and the extracellular and intracellular concentrations of three determining nutrients, that is, free intracellular nitrogen, phosphate, and carbohydrate compounds. Three techniques of global sensitivity analysis are successively applied to assess the model parameter influence and potential correlation. The resulting dynamic model is able to predict plant growth for the two most encountered plant bioprocesses, namely suspension cells and hairy roots.

Distribution of selected virulence genes and antibiotic resistance in Enterococcus species isolated from the South Nation River drainage basin, Ontario, Canada.

AIMS: Isolate and characterize water enterococci from the South Nation River drainage basin, an area dominated by agriculture. METHODS AND RESULTS: A total of 1558 enterococci were isolated from 204 water samples from the South Nation River obtained over a 3-year period. PCR was used to identify isolates to the species level and characterize them for carriage of 12 virulence determinants. Antibiotic resistance was evaluated phenotypically. Enterococcus faecalis (36·4%), Enterococcus faecium (9·3%) and Enterococcus durans (8·5%) were the major enterococci species isolated. Enterococci carrying more than two virulence determinants were more frequently detected in the summer (59·6%) than in other seasons (≤ 37·6%). Very few isolates (≤ 2·0%) were resistant to category I antibiotics ciprofloxacin and vancomycin. CONCLUSIONS: Comparison of major water enterococci species with major faecal enterococci species obtained from various host groups (human, domesticated mammals and birds, wildlife) in this drainage basin suggest that water enterococci may have varied faecal origins. The low level of antibiotic resistance among enterococci suggests that dispersion of antibiotic resistance via waterborne enterococci in this watershed is not significant. SIGNIFICANCE AND IMPACT OF THE STUDY: The data obtained in this study suggests that water enterococci in the SNR have a faecal origin and that their potential impact on public health regarding antibiotic resistance and virulence determinants is minimal.

hnRNP A1 regulates UV-induced NF-kappaB signalling through destabilization of cIAP1 mRNA.

cIAP1 is an important member of the inhibitor of apoptosis family of proteins and is involved in the regulation of the NF-kappaB-signalling pathway downstream of the TNF receptor. We report here that UV irradiation leads to downregulation of cIAP1 expression because of enhanced cIAP1 mRNA destabilization. An AU-rich element located within the 3' untranslated region of cIAP1 mRNA is sufficient to mediate cIAP1 mRNA instability. Furthermore, we have identified hnRNP A1 as a cIAP1 3'UTR-binding protein. hnRNP A1 is a primarily nuclear protein, but accumulates in the cytoplasm after exposure of cells to UV irradiation. Indeed, we find that hnRNP A1 enhances the destabilization of cIAP1 mRNA during UV irradiation. Moreover, siRNA-mediated knockdown of hnRNP A1 restores cIAP1 levels and prevents UV irradiation-induced activation of the NF-kappaB signal transduction pathway, suggesting that hnRNP A1 is an essential post-transcriptional modulator of cIAP1 expression, and thus cIAP1 activity.

A predictive nutritional model for plant cells and hairy roots.

A structured nutritional model is proposed to describe growth and nutritional behavior of Eschscholtzia californica suspension cells and Catharanthus roseus and Daucus carota hairy roots in in vitro culture. The model describes the cells specific growth rate from concentration of intracellular nutrients such as inorganic phosphate (Pi), nitrogen sources (NO(3) (-) and NH(4) (+)) and sugars. Two-level Michaelis-Menten kinetics are used to describe Pi and NO(3) (-) uptake and simple Michaelis-Menten kinetics for description of sugars uptake. Model parameters for each cell line were calibrated using data from batch cultures. The predictive capacity of the model was tested using data from medium exchange hairy root cultures. The model describes growth and nutritional behavior for the cell and hairy root lines. A sensitivity analysis was performed to identify critical model parameters and effect of initial conditions. The cell and hairy roots lines are also compared from their kinetic parameters. The kinetic model is efficient for describing and predicting growth and nutritional behaviors of suspension cells and hairy roots.

Relationship between antioxidant intakes and class I sarcopenia in elderly men and women.

BACKGROUND: The effect of nutritional intake on sarcopenia has been mostly examined in class II sarcopenia, i.e. when muscle mass has sufficiently decreased to induce a loss in physical capacity. Although this provides important information regarding the treatment of sarcopenia, it may not help highlight mechanisms involved at the very beginning of its development. HYPOTHESIS: We hypothesized that class I sarcopenia is associated with differences in antioxidant intakes (vitamins A, C, E and selenium) and status in healthy, older white men and women when physical activity and protein intake are taken into account. DESIGN: Fat-free mass and total appendicular skeletal muscle mass was determined by dual-energy X-ray absorptiometry in 50 healthy, older white men (n = 16) and women (n = 34) aged 60-75 yrs. Physical activity energy expenditure (PAEE) was determined using a Caltrac accelerometer over a 3-d period. Dietary protein and antioxidant intakes were estimated from a 3-d food record and serum total antioxidant activity (TAA) was measured by a ferrylmyoglobin- ABTS assay. RESULTS: The prevalence of class I sarcopenia was 23.5 % in women and 25.0 % in men; 12 participants were thus considered sarcopenic (4 men and 8 women) and 38 participants were considered nonsarcopenic (12 men and 26 women). Our results showed that PAEE, serum albumin concentrations, TAA, and the four antioxidants intake levels were similar between groups. On the other hand, our results showed that total protein intake was significantly higher (P < 0.01) in the non-sarcopenic group than in the sarcopenic group. Also, the number of Recommended Dietary Allowances (RDAs) reached for the antioxidant nutrients and protein intakes by the non-sarcopenic group was significantly higher (P < 0.01) than in the sarcopenic group. CONCLUSIONS: Although there were no significant differences between the sarcopenic and the non-sarcopenic group when antioxidant intakes were considered individually, we observed that the number of RDAs reached for antioxidant micronutrients and protein in healthy, older white men and women was lower in sarcopenic than nonsarcopenic individuals. Our results also suggest that a higher total dietary protein intake is associated with the preservation of muscle mass loss although both groups displayed values above actual RDAs. Obviously, prospective studies are needed to determine the minimum amount of protein in the diet needed to prevent class I sarcopenia and to examine the utility of antioxidant intake to combat the age-related loss in skeletal muscle mass.

Dynamic flux cartography of hairy roots primary metabolism.

A dynamic model for plant cell and hairy root primary metabolism is presented. The model includes nutrient uptake (Pi, sugars, nitrogen sources), the glycolysis and pentose phosphate pathways, the TCA cycle, amino acid biosynthesis, respiratory chain, biosynthesis of cell building blocks (structural hexoses, organic acids, lipids, and organic phosphated molecules). The energy shuttles (ATP, ADP) and cofactors (NAD/H, NADP/H) are also included. The model describes the kinetics of 44 biochemical reactions (fluxes) of the primary metabolism of plant cells and includes 41 biochemical species (metabolites, nutrients, biomass components). Multiple Michaelis-Menten type kinetics are used to describe biochemical reaction rates. Known regulatory phenomena on metabolic pathways are included using sigmoid switch functions. A visualization framework showing fluxes and metabolite concentrations over time is presented. The visualization of fluxes and metabolites is used to analyze simulation results from Catharanthus roseus hairy root 50 d batch cultures. The visualization of the metabolic system allows analyzing split ratios between pathways and flux time-variations. For carbon metabolism, the cells were observed to have relatively high and stable fluxes for the central carbon metabolism and low and variable fluxes for anabolic pathways. For phosphate metabolism, a very high free intracellular Pi turnover rate was observed with higher flux variations than for the carbon metabolism. Nitrogen metabolism also exhibited large flux variations. The potential uses of the model are also discussed.