527 research outputs found

    Glycosylation of hyperthermostable designer cellulosome components yields enhanced stability and cellulose hydrolysis

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    Biomass deconstruction remains integral for enabling second‐generation biofuel production at scale. However, several steps necessary to achieve significant solubilization of biomass, notably harsh pretreatment conditions, impose economic barriers to commercialization. By employing hyperthermostable cellulase machinery, biomass deconstruction can be made more efficient, leading to milder pretreatment conditions and ultimately lower production costs. The hyperthermophilic bacterium Caldicellulosiruptor bescii produces extremely active hyperthermostable cellulases, including the hyperactive multifunctional cellulase CbCel9A/Cel48A. Recombinant CbCel9A/Cel48A components have been previously produced in Escherichia coli and integrated into synthetic hyperthermophilic designer cellulosome complexes. Since then, glycosylation has been shown to be vital for the high activity and stability of CbCel9A/Cel48A. Here, we studied the impact of glycosylation on a hyperthermostable designer cellulosome system in which two of the cellulosomal components, the scaffoldin and the GH9 domain of CbCel9A/Cel48A, were glycosylated as a consequence of employing Ca. bescii as an expression host. Inclusion of the glycosylated components yielded an active cellulosome system that exhibited long‐term stability at 75 °C. The resulting glycosylated designer cellulosomes showed significantly greater synergistic activity compared to the enzymatic components alone, as well as higher thermostability than the analogous nonglycosylated designer cellulosomes. These results indicate that glycosylation can be used as an essential engineering tool to improve the properties of designer cellulosomes. Additionally, Ca. bescii was shown to be an attractive candidate for production of glycosylated designer cellulosome components, which may further promote the viability of this bacterium both as a cellulase expression host and as a potential consolidated bioprocessing platform organism

    Stability of Executive Functioning of Moderately-Late Preterm and Full-Term Born Children at Ages 11 and 19:The TRAILS Cohort Study

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    Moderately-late preterm-born children (MLPs, 32-36 weeks gestational age, GA) have poorer executive functioning (EF) at primary school age than full-term children (FTs). Evidence is lacking on their EF in adolescence, but for early preterm-born children, this has been shown to be much poorer. We, therefore, compared EF of MLPs and FTs at ages 11 and 19 and assessed development between these ages. We obtained data from TRAILS, a community-based prospective cohort study in the northern Netherlands, on 98 MLPs and 1832 FTs. We assessed EF by the Amsterdam Neuropsychological Tasks (ANT) at ages 11 and 19 years and computed gender-specific z-scores on reaction time and accuracy. We compared baseline speed, pattern search, working memory, sustained attention, inhibition, and attentional flexibility of MLPs and FTs crude, and adjusted for small-for-GA status, socioeconomic status, and estimated intelligence. MLPs and FTs performed similarly on all EF components at ages 11 and 19, except for the speed, but not the accuracy measure of attentional flexibility. This was slightly poorer for MLPs than FTs at age 19 (adjusted B 0.25; 95% confidence interval: 0.00 to 0.50; p = 0.047), but not at age 11 (adjusted B -0.02; -0.19 to 0.22; p = 0.87). Differences in EF between MLPs and FTs did not change significantly from age 11 to 19. MLPs had comparable EF on most components as FTs, with only attentional flexibility at age 19 developing slightly poorer for MLPs than for FTs. These findings suggest the effects of MLP birth on long-term EF to be small

    Comparing Outcomes for Community-Acquired Pneumonia Between Females and Males: Results from the University of Louisville Pneumonia Study

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    Introduction: Male sex is currently considered to be a risk factor for worsened community-acquired pneumonia (CAP) outcomes compared to female sex; hence, female sex equates to a lower score on the Pneumonia Severity Index. There is no recent update on sex-based outcomes of patients with CAP. The objective of this study was to compare the outcomes of CAP between females and males. Methods: This was a secondary analysis of the University of Louisville Pneumonia Study database. It was a prospective population-based cohort study of all hospitalized adults with CAP who were residents of Jefferson County in the city of Louisville, Kentucky. The study included data from June 1, 2014, to May 31, 2016, and data from October 1, 2016, to May 31, 2017. The study population was divided into two groups: females and males. Results: Female patients had a 13% lower mortality at one year compared to males (aHR 1.13 [95% CI 1.05–1.23], P=0.002). There was no significant difference in mortality between the two groups during hospitalization or at 30-day or six-month follow-up. The median time to discharge for both female and male patients hospitalized with CAP was five days (interquartile range [IQR] 3, 9 days). The median time to clinical stability for both female and male patients hospitalized with CAP was two days (IQR 1, 4 days). Conclusion: This study shows that female patients had significantly lower one-year mortality compared to males. There was no significant difference between females and males in time to clinical stability or length of stay. Further investigation is needed to examine whether risk factors associated with female and male sex predict outcomes among hospitalized patients due to CAP

    Differential loss of effector genes in three recently expanded pandemic clonal lineages of the rice blast fungus

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    BACKGROUND: Understanding the mechanisms and timescales of plant pathogen outbreaks requires a detailed genome-scale analysis of their population history. The fungus Magnaporthe (Syn. Pyricularia) oryzae-the causal agent of blast disease of cereals- is among the most destructive plant pathogens to world agriculture and a major threat to the production of rice, wheat, and other cereals. Although M. oryzae is a multihost pathogen that infects more than 50 species of cereals and grasses, all rice-infecting isolates belong to a single genetically defined lineage. Here, we combined the two largest genomic datasets to reconstruct the genetic history of the rice-infecting lineage of M. oryzae based on 131 isolates from 21 countries. RESULTS: The global population of the rice blast fungus consists mainly of three well-defined genetic groups and a diverse set of individuals. Multiple population genetic tests revealed that the rice-infecting lineage of the blast fungus probably originated from a recombining diverse group in Southeast Asia followed by three independent clonal expansions that took place over the last ~ 200 years. Patterns of allele sharing identified a subpopulation from the recombining diverse group that introgressed with one of the clonal lineages before its global expansion. Remarkably, the four genetic lineages of the rice blast fungus vary in the number and patterns of presence and absence of candidate effector genes. These genes encode secreted proteins that modulate plant defense and allow pathogen colonization. In particular, clonal lineages carry a reduced repertoire of effector genes compared with the diverse group, and specific combinations of presence and absence of effector genes define each of the pandemic clonal lineages. CONCLUSIONS: Our analyses reconstruct the genetic history of the rice-infecting lineage of M. oryzae revealing three clonal lineages associated with rice blast pandemics. Each of these lineages displays a specific pattern of presence and absence of effector genes that may have shaped their adaptation to the rice host and their evolutionary history

    Synergistic effect of methyljasmonate and cyclodextrin on stilbene biosynthesis pathway gene expression and resveratrol production in Monastrell grapevine cell cultures

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    <p>Abstract</p> <p>Background</p> <p>Plant cell cultures have been shown as feasible systems for the production of secondary metabolites, being the elicitation with biotic or abiotic stimuli the most efficient strategy to increase the production of those metabolites. Vitaceae phytoalexins constitute a group of molecules belonging to the stilbene family which are derivatives of the <it>trans</it>-resveratrol structure and are produced by plants and cell cultures as a response to biotic and abiotic stresses. The potential benefits of resveratrol on human health have made it one of the most thoroughly studied phytochemical molecules. The aim of this study was to evaluate the elicitor effect of both cyclodextrin (CD) and methyljasmonate (MeJA) on grapevine cell cultures by carrying out a quantitative analysis of their role on resveratrol production and on the expression of stilbene biosynthetic genes in <it>Vitis vinifera </it>cv Monastrell albino cell suspension cultures.</p> <p>Findings</p> <p>MeJA and CD significantly but transiently induced the expression of stilbene biosynthetic genes when independently used to treat grapevine cells. This expression correlated with resveratrol production in CD-treated cells but not in MeJA-treated cells, which growth was drastically affected. In the combined treatment of CD and MeJA cell growth was similarly affected, however resveratrol production was almost one order of magnitude higher, in correlation with maximum expression values for stilbene biosynthetic genes.</p> <p>Conclusion</p> <p>The effect of MeJA on cell division combined with a true and strong elicitor like CD could be responsible for the observed synergistic effect of both compounds on resveratrol production and on the expression of genes in the stilbene pathway.</p

    A generalized conformational energy function of DNA derived from molecular dynamics simulations

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    Proteins recognize DNA sequences by two different mechanisms. The first is direct readout, in which recognition is mediated by direct interactions between the protein and the DNA bases. The second is indirect readout, which is caused by the dependence of conformation and the deformability of the DNA structure on the sequence. Various energy functions have been proposed to evaluate the contribution of indirect readout to the free-energy changes in complex formations. We developed a new generalized energy function to estimate the dependence of the deformability of DNA on the sequence. This function was derived from molecular dynamics simulations previously conducted on B-DNA dodecamers, each of which had one possible tetramer sequence embedded at its center. By taking the logarithm of the probability distribution function (PDF) for the base-step parameters of the central base-pair step of the tetramer, its ability to distinguish the native sequence from random ones was superior to that with the previous method that approximated the energy function in harmonic form. From a comparison of the energy profiles calculated with these two methods, we found that the harmonic approximation caused significant errors in the conformational energies of the tetramers that adopted multiple stable conformations

    Glycosylation of hyperthermostable designer cellulosome components yields enhanced stability and cellulose hydrolysis

    Get PDF
    Biomass deconstruction remains integral for enabling second‐generation biofuel production at scale. However, several steps necessary to achieve significant solubilization of biomass, notably harsh pretreatment conditions, impose economic barriers to commercialization. By employing hyperthermostable cellulase machinery, biomass deconstruction can be made more efficient, leading to milder pretreatment conditions and ultimately lower production costs. The hyperthermophilic bacterium Caldicellulosiruptor bescii produces extremely active hyperthermostable cellulases, including the hyperactive multifunctional cellulase CbCel9A/Cel48A. Recombinant CbCel9A/Cel48A components have been previously produced in Escherichia coli and integrated into synthetic hyperthermophilic designer cellulosome complexes. Since then, glycosylation has been shown to be vital for the high activity and stability of CbCel9A/Cel48A. Here, we studied the impact of glycosylation on a hyperthermostable designer cellulosome system in which two of the cellulosomal components, the scaffoldin and the GH9 domain of CbCel9A/Cel48A, were glycosylated as a consequence of employing Ca. bescii as an expression host. Inclusion of the glycosylated components yielded an active cellulosome system that exhibited long‐term stability at 75 °C. The resulting glycosylated designer cellulosomes showed significantly greater synergistic activity compared to the enzymatic components alone, as well as higher thermostability than the analogous nonglycosylated designer cellulosomes. These results indicate that glycosylation can be used as an essential engineering tool to improve the properties of designer cellulosomes. Additionally, Ca. bescii was shown to be an attractive candidate for production of glycosylated designer cellulosome components, which may further promote the viability of this bacterium both as a cellulase expression host and as a potential consolidated bioprocessing platform organism
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