In vivo evolution of lactic acid hyper-tolerant Clostridium thermocellum

Lactic acid (LA) has several applications in the food, cosmetics and pharmaceutical industries, as well as in the production of biodegradable plastic polymers, namely polylactides. Industrial production of LA is essentially based on microbial fermentation. Recent reports have shown the potential of the cellulolytic bacterium Clostridium thermocellum for direct LA production from inexpensive lignocellulosic biomass. However, C. thermocellum is highly sensitive to acids and does not grow at pH \u3c 6.0. Improvement of LA tolerance of this microorganism is pivotal for its application in cost-efficient production of LA. In the present study, the LA tolerance of C. thermocellum strains LL345 (wild-type fermentation profile) and LL1111 (high LA yield) was increased by adaptive laboratory evolution. At large inoculum size (10 %), the maximum tolerated LA concentration of strain LL1111 was more than doubled, from 15 g/L to 35 g/L, while subcultures evolved from LL345 showed 50–85 % faster growth in medium containing 45 g/L LA. Gene mutations (pyruvate phosphate dikinase, histidine protein kinase/phosphorylase) possibly affecting carbohydrate and/or phosphate metabolism have been detected in most LA-adapted populations. Although improvement of LA tolerance may sometimes also enable higher LA production in microorganisms, C. thermocellum LA-adapted cultures showed a yield of LA, and generally of other organic acids, similar to or lower than parental strains. Based on its improved LA tolerance and LA titer similar to its parent strain (LL1111), mixed adapted culture LL1630 showed the highest performing phenotype and could serve as a framework for improving LA production by further metabolic engineering

A role for eisosomes in maintenance of plasma membrane phosphoinositide levels

The plasma membrane delineates the cell and mediates its communication and material exchange with the environment. Many processes of the plasma membrane occur through interactions of proteins with phosphatidylinositol(4,5)-bisphosphate (PI(4,5)P2), which is highly enriched in this membrane and is a key determinant of its identity. Eisosomes function in lateral organization of the plasma membrane, but the molecular function of their major protein subunits, the BAR domain–containing proteins Pil1 and Lsp1, is poorly understood. Here we show that eisosomes interact with the PI(4,5)P2 phosphatase Inp51/Sjl1, thereby recruiting it to the plasma membrane. Pil1 is essential for plasma membrane localization and function of Inp51 but not for the homologous phosphatidylinositol bisphosphate phosphatases Inp52/Sjl2 and Inp53/Sjl3. Consistent with this, absence of Pil1 increases total and available PI(4,5)P2 levels at the plasma membrane. On the basis of these findings, we propose a model in which the eisosomes function in maintaining PI(4,5)P2 levels by Inp51/Sjl1 recruitment

Driven vortices in 3D layered superconductors: Dynamical ordering along the c-axis

We study a 3D model of driven vortices in weakly coupled layered superconductors with strong pinning. Above the critical force $F_c$, we find a plastic flow regime in which pancakes in different layers are uncoupled, corresponding to a pancake gas. At a higher $F$, there is an ``smectic flow'' regime with short-range interlayer order, corresponding to an entangled line liquid. Later, the transverse displacements freeze and vortices become correlated along the c-axis, resulting in a transverse solid. Finally, at a force $F_s$ the longitudinal displacements freeze and we find a coherent solid of rigid lines.Comment: 4 pages, 3 postscript figure

A global transcriptional network connecting noncoding mutations to changes in tumor gene expression.

Although cancer genomes are replete with noncoding mutations, the effects of these mutations remain poorly characterized. Here we perform an integrative analysis of 930 tumor whole genomes and matched transcriptomes, identifying a network of 193 noncoding loci in which mutations disrupt target gene expression. These 'somatic eQTLs' (expression quantitative trait loci) are frequently mutated in specific cancer tissues, and the majority can be validated in an independent cohort of 3,382 tumors. Among these, we find that the effects of noncoding mutations on DAAM1, MTG2 and HYI transcription are recapitulated in multiple cancer cell lines and that increasing DAAM1 expression leads to invasive cell migration. Collectively, the noncoding loci converge on a set of core pathways, permitting a classification of tumors into pathway-based subtypes. The somatic eQTL network is disrupted in 88% of tumors, suggesting widespread impact of noncoding mutations in cancer

The Exometabolome of Clostridium Thermocellum Reveals Overflow Metabolism at High Cellulose Loading

BackgroundClostridium thermocellum is a model thermophilic organism for the production of biofuels from lignocellulosic substrates. The majority of publications studying the physiology of this organism use substrate concentrations of ≤10 g/L. However, industrially relevant concentrations of substrate start at 100 g/L carbohydrate, which corresponds to approximately 150 g/L solids. To gain insight into the physiology of fermentation of high substrate concentrations, we studied the growth on, and utilization of high concentrations of crystalline cellulose varying from 50 to 100 g/L by C. thermocellum.

The Exometabolome of Clostridium Thermocellum Reveals Overflow Metabolism at High Cellulose Loading

BackgroundClostridium thermocellum is a model thermophilic organism for the production of biofuels from lignocellulosic substrates. The majority of publications studying the physiology of this organism use substrate concentrations of ≤10 g/L. However, industrially relevant concentrations of substrate start at 100 g/L carbohydrate, which corresponds to approximately 150 g/L solids. To gain insight into the physiology of fermentation of high substrate concentrations, we studied the growth on, and utilization of high concentrations of crystalline cellulose varying from 50 to 100 g/L by C. thermocellum.

Semi-Classical Description of Antiproton Capture on Atomic Helium

A semi-classical, many-body atomic model incorporating a momentum-dependent Heisenberg core to stabilize atomic electrons is used to study antiproton capture on Helium. Details of the antiproton collisions leading to eventual capture are presented, including the energy and angular momentum states of incident antiprotons which result in capture via single or double electron ionization, i.e. into [He$^{++}\,\bar p$ or He$^{+}\,\bar p$], and the distribution of energy and angular momentum states following the Auger cascade. These final states are discussed in light of recently reported, anomalously long-lived antiproton states observed in liquid He.Comment: 15 pages, 9 figures may be obtained from authors, Revte

Sibling relationships and family functioning in siblings of early adolescents, adolescents and young adults with autism spectrum disorder

The purpose of the study was to investigate how family functioning (defined as the ability that family members hold to manage stressful events, and intimate and social relationships), the degree to which family members feel happy and fulfilled with each other (called family satisfaction), and the demographical characteristics of siblings (age and gender) impacted on sibling relationships. The Circumplex Model of Marital and Family Systems and Behavioral Systems constituted the theoretical frameworks that guided our study. Eighty-six typically developing adolescents and young adults having a sister or a brother with autism spectrum disorder were enrolled. Results indicated that the youngest age group (early adolescents) reported to engage more frequently in negative behaviors with their siblings with ASD than the two older age groups (middle adolescents and young adults). No significant differences were found among the three age groups regarding behaviors derived from attachment, caregiving and affiliative systems. Family satisfaction and age significantly predicted behaviors during sibling interactions. Suggestions on prevention and intervention programs were discussed in order to prevent parentification among typically developing siblings and decrease episodes of quarrels and overt conflicts between brothers and sisters with and without AS

Length of the weaning period affects postweaning growth, health, and carcass merit of ranch-direct beef calves weaned during the fall

Bovine respiratory disease (BRD) is the most economically devastating feedlot disease. Risk factors associated with incidence of BRD include (1) stress associated with maternal separation, (2) stress associated with introduction to an unfamiliar environment, (3) poor intake associated with introduction of novel feedstuffs into the animal\u27s diet, (4) exposure to novel pathogens upon transport to a feeding facility and commingling with unfamiliar cattle, (5) inappropriately administered respiratory disease vaccination programs, and (6) poor response to respiratory disease vaccination programs. Management practices that are collectively referred to as preconditioning are thought to minimize damage to the beef carcass from the BRD complex. Preconditioning management reduces the aforementioned risk factors for respiratory disease by (1) using a relatively long ranch-of-origin weaning period following maternal separation, (2) exposing calves to concentrate-type feedstuffs, and (3) producing heightened resistance to respiratory disease-causing organisms through a preweaning vaccination program. The effectiveness of such programs for preserving animal performance is highly touted by certain segments of the beef industry. Ranch-of-origin weaning periods of up to 60 days are suggested for preconditioning beef calves prior to sale; however, optimal length of the ranch-of-origin weaning period has not been determined experimentally. The objective of this study was to test the validity of beef industry assumptions about appropriate length of ranch-of-origin weaning periods for calves aged 160 to 220 days and weaned during the fall

Radial Construction of an Arterial Wall

SummarySome of the most serious diseases involve altered size and structure of the arterial wall. Elucidating how arterial walls are built could aid understanding of these diseases, but little is known about how concentric layers of muscle cells and the outer adventitial layer are assembled and patterned around endothelial tubes. Using histochemical, clonal, and genetic analysis in mice, here we show that the pulmonary artery wall is constructed radially, from the inside out, by two separate but coordinated processes. One is sequential induction of successive cell layers from surrounding mesenchyme. The other is controlled invasion of outer layers by inner layer cells through developmentally regulated cell reorientation and radial migration. We propose that a radial signal gradient controls these processes and provide evidence that PDGF-B and at least one other signal contribute. Modulation of such radial signaling pathways may underlie vessel-specific differences and pathological changes in arterial wall size and structure.Video Abstrac