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Background and Hypothesis Risk for cannabis use and schizophrenia is influenced in part by genetic factors, and there is evidence that genetic risk for schizophrenia is associated with subclinical psychotic-like experiences (PLEs). Few studies to date have examined whether genetic risk for schizophrenia is associated with cannabis-related PLEs. Study Design We tested whether measures of cannabis involvement and polygenic risk scores (PRS) for schizophrenia were associated with self-reported cannabis-related experiences in a sample ascertained for alcohol use disorders (AUDs), the Collaborative Study on the Genetics of Alcoholism (COGA). We analyzed 4832 subjects (3128 of European ancestry and 1704 of African ancestry; 42% female; 74% meeting lifetime criteria for an AUD). Study Results Cannabis use disorder (CUD) was prevalent in this analytic sample (70%), with 40% classified as mild, 25% as moderate, and 35% as severe. Polygenic risk for schizophrenia was positively associated with cannabis-related paranoia, feeling depressed or anhedonia, social withdrawal, and cognitive difficulties, even when controlling for duration of daily cannabis use, CUD, and age at first cannabis use. The schizophrenia PRS was most robustly associated with cannabis-related cognitive difficulties (β = 0.22, SE = 0.04, P = 5.2e−7). In an independent replication sample (N = 1446), associations between the schizophrenia PRS and cannabis-related experiences were in the expected direction and not statistically different in magnitude from those in the COGA sample. Conclusions Among individuals who regularly use cannabis, genetic liability for schizophrenia—even in those without clinical features—may increase the likelihood of reporting unusual experiences related to cannabis use.

Attempts to identify opioid users with increased risk of escalating to opioid use disorder (OUD) have had limited success. Retrospectively assessed subjective effects of initial opioid misuse were compared in a pilot sample of opioid misusers (nonmedical use ≤60 times lifetime) who had never met criteria for OUD (N = 14) and heroin-addicted individuals in treatment for OUD (N = 15). Relative to opioid misusers without a lifetime OUD diagnosis, individuals with OUD reported greater euphoria and other positive emotions, activation, pruritus, and internalizing symptoms. Consistent with these findings, proxy Addiction Research Center Inventory (ARCI) Amphetamine Group, and Morphine Benzedrine Group scale mean item scores were significantly higher in those with OUD. Replication was attempted in opioid misusers with (N = 25) and without OUD (N = 25) who were assessed as part of an ongoing genetic study. We observed similar significant between-group differences in individual subjective effect items and ARCI scale mean item scores in the replication sample. We, thus confirm findings from prior reports that retrospectively assessed subjective responses to initial opioid exposure differ significantly between opioid users who do, and do not, progress to OUD. Our report extends these findings in comparisons limited to opioid misusers. Additional research will be necessary to examine prospectively whether the assessment of subjective effects after initial use has predictive utility in the identification of individuals more likely to progress to OUD.

Opioid addiction (OA) has strong heritability, yet few genetic variant associations have been robustly identified. Only rs1799971, the A118G variant inOPRM1, has been identified as a genome-wide significant association with OA and independently replicated. We applied genomic structural equation modeling to conduct a GWAS of the new Genetics of Opioid Addiction Consortium (GENOA) data and published studies (Psychiatric Genomics Consortium, Million Veteran Program, and Partners Health), comprising 23,367 cases and effective sample size of 88,114 individuals of European ancestry. Genetic correlations among the various OA phenotypes were uniformly high (rg> 0.9). We observed the strongest evidence to date forOPRM1: lead SNP rs9478500 (p=2.56×10−9). Gene-based analyses identified novel genome-wide significant associations withPPP6CandFURIN. Variants within these loci appear to be pleiotropic for addiction and related traits.

Background Scheffersomyces stipitis is an important yeast species in the field of biorenewables due to its desired capacity for xylose utilization. It has been recognized that redox balance plays a critical role in S. stipitis due to the different cofactor preferences in xylose assimilation pathway. However, there has not been any systems level understanding on how the shift in redox balance contributes to the overall metabolic shift in S. stipitis to cope with reduced oxygen uptake. Genome-scale metabolic network models (GEMs) offer the opportunity to gain such systems level understanding; however, currently the two published GEMs for S. stipitis cannot be used for this purpose, as neither of them is able to capture the strain’s fermentative metabolism reasonably well due to their poor prediction of xylitol production, a key by-product under oxygen limited conditions. Results A system identification-based (SID-based) framework that we previously developed for GEM validation is expanded and applied to refine a published GEM for S. stipitis, iBB814. After the modified GEM, named iDH814, was validated using literature data, it is used to obtain genome-scale understanding on how redox cofactor shifts when cells respond to reduced oxygen supply. The SID-based framework for GEM analysis was applied to examine how the environmental perturbation (i.e., reduced oxygen supply) propagates through the metabolic network, and key reactions that contribute to the shifts of redox and metabolic state were identified. Finally, the findings obtained through GEM analysis were validated using transcriptomic data. Conclusions iDH814, the modified model, was shown to offer significantly improved performance in terms of matching available experimental results and better capturing available knowledge on the organism. More importantly, our analysis based on iDH814 provides the first genome-scale understanding on how redox balance in S. stipitis was shifted as a result of reduced oxygen supply. The systems level analysis identified the key contributors to the overall metabolic state shift, which were validated using transcriptomic data. The analysis confirmed that S. stipitis uses a concerted approach to cope with the stress associated with reduced oxygen supply, and the shift of reducing power from NADPH to NADH seems to be the center theme that directs the overall shift in metabolic states. Electronic supplementary material The online version of this article (10.1186/s12934-018-0983-y) contains supplementary material, which is available to authorized users.

Lipid production by oleaginous yeasts is optimal at high carbon-to-nitrogen ratios. In the current study, nitrogen and carbon consumption by Lipomyces starkeyi were directly measured in defined minimal media with nitrogen content and agitation rates as variables. Shake flask cultures with an initial C:N ratio of 72:1 cultivated at 200rpm resulted in a lipid output of 10g/L, content of 55%, yield of 0.170g/g, and productivity of 0.06g/L/h. All of these values decreased by ≈50-60% when the agitation rate was raised to 300rpm or when the C:N ratio was lowered to 24:1, demonstrating the importance of these parameters. Under all conditions, L. starkeyi cultures tolerated acidified media (pH≈2.6) without difficulty, and produced considerable amounts of alcohols; including ethanol, mannitol, arabitol, and 2,3-butanediol. L. starkeyi also produced lipids from a corn stover hydrolysate, showing its potential to produce biofuels from renewable agricultural feedstocks.

Yeasts are unicellular fungi that do not form fruiting bodies.Although the yeast lifestyle has evolved multiple times, mostknown species belong to the subphylum Saccharomycotina(syn. Hemiascomycota, hereafter yeasts). This diverse groupincludes the premier eukaryotic model system,Saccharomyces cerevisiae; the common human commensaland opportunistic pathogen, Candida albicans; and over1000 other known species (with more continuing to bediscovered). Yeasts are found in every biome and continent andare more genetically diverse than angiosperms or chordates.Ease of culture, simple life cycles, and small genomes (10?20 Mbp) have made yeasts exceptional models for moleculargenetics, biotechnology, and evolutionary genomics. Herewe discuss recent developments in understanding thegenomic underpinnings of the making of yeast biodiversity,comparing and contrasting natural and human-associatedevolutionary processes. Only a tiny fraction of yeastbiodiversity and metabolic capabilities has been tapped byindustry and science. Expanding the taxonomic breadth ofdeep genomic investigations will further illuminate how genomefunction evolves to encode their diverse metabolisms andecologies. Fil: Hittinger, Chris Todd. University Of Wisconsin; Estados Unidos. University Of Wisconsin; Estados Unidos Fil: Rokas, Antonis. Vanderbilt University; Estados Unidos Fil: Bai, Feng Yan. Chinese Academy Of Sciences; República de China Fil: Boekhout, Teun. Chinese Academy Of Sciences; República de China. CBS-KNAW Fungal Biodiversity Centre; Países Bajos. Second Military Medical University; China Fil: Gonçalves, Paula. Universidade Nova de Lisboa; Portugal Fil: Jeffries, Thomas W.. University Of Wisconsin; Estados Unidos Fil: Kominek, Jacek. University Of Wisconsin; Estados Unidos Fil: Lachance, Marc Andre. University of Western Ontario; Canadá Fil: Libkind Frati, Diego. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Patagonia Norte. Instituto de Investigación en Biodiversidad y Medioambiente; Argentina. Universidad Nacional del Comahue; Argentina Fil: Rosa, Carlos A.. Universidade Federal do Minas Gerais; Brasil Fil: Sampaio, José Paulo. Universidade Nova de Lisboa; Portugal Fil: Kurtzman, Cletus P.. United States Department of Agriculture; Argentina

Background Recovery has become a central concept in mental health service delivery, and several recovery-focused measures exist for adults. The concept's applicability to young people's mental health experience has been neglected, and no measures yet exist. Aim The aim of this work is to develop measures of recovery for use in specialist child and adolescent mental health services. Method On the basis of 21 semi-structured interviews, three recovery measures were devised, one for completion by the young person and two for completion by the parent/carer. Two parent/carer measures were devised in order to assess both their perspective on their child's recovery and their own recovery process. The questionnaires were administered to a UK sample of 47 young people (10–18 years old) with anxiety and depression and their parents, along with a measure used to routinely assess treatment progress and outcome and a measure of self-esteem. Results All three measures had high internal consistency (alpha ≥ 0.89). Young people's recovery scores were correlated negatively with scores on a measure used to routinely assess treatment progress and outcome (r = −0.75) and positively with self-esteem (r = 0.84). Parent and young persons' reports of the young person's recovery were positively correlated (r = 0.61). Parent report of the young person's recovery and of their own recovery process were positively correlated (r = 0.75). Conclusion The three measures have the potential to be used in mental health services to assess recovery processes in young people with mental health difficulties and correspondence with symptomatic improvement. The measures provide a novel way of capturing the parental/caregiver perspective on recovery and caregivers' own wellbeing. Copyright © 2014 John Wiley & Sons, Ltd. Key Practitioner Message •No tools exist to evaluate recovery-relevant processes in young people treated in specialist mental health services. •This study reports on the development and psychometric evaluation of three self-report recovery-relevant assessments for young people and their caregivers. •Findings indicate a high degree of correspondence between young person and caregiver reports of recovery in the former. •The recovery assessments correlate inversely with a standardized symptom-focused measure and positively with self-esteem.

This study employed cell recycling, batch adaptation, cell mating and high-throughput screening to select adapted Spathaspora passalidarum strains with improved fermentative ability. The most promising candidate YK208-E11 (E11) showed a 3-fold increase in specific fermentation rate compared to the parental strain and an ethanol yield greater than 0.45 g/g substrate while co-utilizing cellobiose, glucose and xylose. Further characterization showed that strain E11 also makes 40% less biomass compared to the parental strain when cultivated in rich media under aerobic conditions. A tetrazolium agar overlay assay in the presence of respiration inhibitors, including rotenone, antimycin A, KCN and salicylhydroxamic acid elucidated the nature of the mutational events. Results indicated that E11 has a deficiency in its respiration system that could contribute to its low cell yield. Strain E11 was subjected to whole genome sequencing and an ∼11 kb deletion was identified; the open reading frames absent in strain E11 code for proteins with predicted functions in respiration, cell division and the actin cytoskeleton, and may contribute to the observed physiology of the adapted strain. Results of the tetrazolium overlay also suggest that cultivation on xylose affects the respiration capacity in the wild-type strain, which could account for its faster fermentation of xylose as compared to glucose. These results support our previous finding that S. passalidarum has highly unusual physiological responses to xylose under oxygen limitation.

Biomass pretreatment is essential to overcome recalcitrance of lignocellulose for ethanol production. In the present study we pretreated giant reed (Arundo donax L.), a perennial, rhizomatous lignocellulosic grass with dilute oxalic acid. The effects of temperature (170–190 °C), acid loading (2–10% w/w) and reaction time (15–40 min) were handled as a single parameter, combined severity. We explored the change in hemicellulose, cellulose and lignin composition following pretreatment and glucan conversion after enzymatic hydrolysis of the solid residue. Two different yeast strains, Scheffersomyces (Pichia) stipitis CBS 6054, which is a native xylose and cellobiose fermenter, and Saccharomyces carlsbergensis FPL-450, which does not ferment xylose or cellobiose, were used along with commercial cellulolytic enzymes in simultaneous saccharification and fermentation (SSF). S. carlsbergensis attained a maximum ethanol concentration of 15.9 g/l after 48 h at pH 5.0, while S. stipitis, at the same condition, took 96 h to reach a similar ethanol value; increasing the pH to 6.0 reduced the S. stipitis lag phase and attained 18.0 g/l of ethanol within 72 h.

Estrogen has been shown to increase endothelial cell (EC) production of nitric oxide (NO) by the rapid activation of endothelial nitric oxide synthase (eNOS). To better understand the mechanisms by which estrogen acutely increases endothelial NO production, proteins of the mitogen activated protein kinase (MAPK) family, ERK, JNK, and p38 were examined to determine their potential role in this process. Bovine aortic endothelial cells (BAEC) were grown to confluence in phenol red-free DMEM F-12, supplemented with 10% fetal calf serum, on 35 cm2 dishes. After confluence, cells were starved for 12–24 hours and then exposed to 10 nM 17β-estradiol (E2) for 1–60 minutes. The cells were then lysed and proteins resolved by SDS-PAGE followed by transfer to nitrocellulose membranes. Membranes were then probed with anti-phosphospecific antibodies for ERK, JNK, and p38. Antibodies specific for total ERK and JNK were used to assure even protein loading. The level of ERK phosphorylation rapidly increased in biphasic pattern after E2 exposure: 1 min, 4.3 fold; 5 min, 1 fold; and 10 min, 7.3 fold (max). However, the level of JNK phosphorylation did not increase throughout the time course. The level of p38 phosphorylation increased rapidly to 3.6 fold at 10 minutes. We demonstrate that E2 rapidly activates ERK in a biphasic manner and p38 in a monophasic manner, but has no effect on the activity of JNK. Our data suggest that rapid eNOS activation by estrogen in vascular endothelial cells is predominantly mediated through ERK and p38 pathway. Rapid activation of ERK and p38 resulting in NO production may substantially contribute to the atheroprotective effect of estrogen.

This research examined cellulolytic effects of fungi and other microbes present in cured sausages on the strength and stability of regenerated cellulose casings (RCC) used in the sausage industry. Occasionally during the curing process, RCC would split or fail, thereby leading to loss of product. The fungus Penicillium sp. BT-F-1, which was isolated from fermented sausages, and other fungi, which were introduced to enable the curing process, produced small amounts of cellulases on RCC in both liquid and solid cultivations. During continued incubation for 15-60 days in solid substrate cultivation (SSC) on RCC support, the fungus Penicillium sp isolate BT-F-1 degraded the casings' dry weights by 15-50% and decreased their tensile strengths by ~75%. Similarly commercial cellulase(s) resulted in 20-50% degradation of RCC in 48 h. During incubation with Penicillium sp BT-F-1, the surface structure of RCC collapsed, resulting in loss of strength and stability of casings. The matrix of industrial RCC comprised 88-93% glucose polymer residues with 0.8-4% xylan impurities. Premature casing failure appeared to result from operating conditions in the manufacturing process that allowed xylan to build up in the extrusion bath. The sausage fungus Penicillium sp BT-F-1 produced xylanases to break down soft xylan pockets prior to slow cellulosic dissolution of RCC.

AMPA receptors mediate fast excitatory transmission in the brain. Neuronal AMPA receptors comprise GluA pore-forming principal subunits and can associate with multiple modulatory components, including transmembrane AMPA receptor regulatory proteins (TARPs) and CNIHs (cornichons). AMPA receptor potentiators and non-competitive antagonists represent potential targets for a variety of neuropsychiatric disorders. Previous studies showed that the AMPA receptor antagonist GYKI-53655 displaces binding of a potentiator from brain receptors but not from recombinant GluA subunits. Here, we asked whether AMPA receptor modulatory subunits might resolve this discrepancy. We find that the cerebellar TARP, stargazin (γ-2), enhances the binding affinity of the AMPA receptor potentiator [(3)H]-LY450295 and confers sensitivity to displacement by non-competitive antagonists. In cerebellar membranes from stargazer mice, [(3)H]-LY450295 binding is reduced and relatively resistant to displacement by non-competitive antagonists. Coexpression of AMPA receptors with CNIH-2, which is expressed in the hippocampus and at low levels in the cerebellar Purkinje neurons, confers partial sensitivity of [(3)H]-LY450295 potentiator binding to displacement by non-competitive antagonists. Autoradiography of [(3)H]-LY450295 binding to stargazer and γ-8-deficient mouse brain sections, demonstrates that TARPs regulate the pharmacology of allosteric AMPA potentiators and antagonists in the cerebellum and hippocampus, respectively. These studies demonstrate that accessory proteins define AMPA receptor pharmacology by functionally linking allosteric AMPA receptor potentiator and antagonist sites.

Corn stover that had been treated with vapor-phase diethyl oxalate released a mixture of mono- and oligosaccharides consisting mainly of xylose and glucose. Following overliming and neutralization, a D-xylulokinase mutant of Pichia stipitis, FPL-YS30 (xyl3-∆1), converted the stover hydrolysate into xylitol. This research examined the effects of phosphoric or gluconic acids used for neutralization and urea or ammonium sulfate used as nitrogen sources. Phosphoric acid improved color and removal of phenolic compounds. D-Gluconic acid enhanced cell growth. Ammonium sulfate increased cell yield and maximum specific cell growth rate independently of the acid used for neutralization. The highest xylitol yield (0.61 g(xylitol)/g(xylose)) and volumetric productivity (0.18 g(xylitol)/g(xylose )l) were obtained in hydrolysate neutralized with phosphoric acid. However, when urea was the nitrogen source the cell yield was less than half of that obtained with ammonium sulfate.

이 연구에서는 백합나무(Liriodendron tulipifera)를 옥살산으로 전처리한 시료로부터 에탄올 생산 가능성을 조사하고, response surface methodology(RSM)를 도입하여 전처리 조건을 분석하고자 한다. 산농도, 전처리 시간, 반응 온도를 조절하여 2(3) factorial central composite experimental design을 바탕으로 각기 다른 l5가지의 전처리 조건에서 시험하였다(central point에서 2반복). 전처리 후 고체 시료는 발효 균주인 Pichia stipitis를 사용하여 동시 당화발효로 에탄올 생산에 이용되었으며, 각각의 시료에서의 72시간에서의 에탄올 생산량(y,g/l)이 최대값으로, 종속변수로써 RSM에 적용되었다. 180℃에서 40분간 0.013 g/ g의 옥살산으로 처리한 시료가 가장 많은 양의 에탄올(9.7 g/l)을 생산하였으며, response surface methodology 분석에 따르면, 전처리 조건에서 온도 인자가 ethanol에 가장 큰 영향을 미치는 것으로 제시되었으며, 결과는 수식화되어 나타내었다.

In this study, we evaluated optimal conditions for ethanol production of the spruce hydrolysate (SH) obtained from diethyl oxalate pretreatment. Fermentable sugar concentration in SH was 29.04 g/l except arabinose. Monosaccharides obtained from the oligomer degradation were mainly mannose (39.26 g/l) and galactose (12.83 g/l). Concentration of 5-HMF and furfural which are inhibitors on ethanol fermentation were 0.09 g/l and 0.04 g/ respectively. Concentration of acetic acid and total phenolic compounds in SH were 1.4 g/l and 2.83 g/. Ethanol production using hydrolysate was 11.7 g/l at optimal pH 6.0 after 48 h. Specific ethanol production was 0.15 (g/(l*h)) at pH 5.0 and 5.5. while that was 0.24 (g/(l*h)) at pH 6.0. Specific ethanol production has difference depend on initial pH for fermentation. Ethanol production was 14.3 g/l after 48 h when xylanase 20 IU was added in SH for degradation of oligomer during fermentation. It implied that ethanol production increased by 22.2% compare with control (without xylanase).

Aqueous dilute acid pretreatments of corncob were conducted using cylindrical pressure vessels in an oil bath. Pretreatments were conducted in a temperature range of 160-190 °C with acid-solution-to-solid-corncob ratio of 2. The acid concentration (v/v) in the pretreatment solution was varied from 0% to 0.7%, depending on temperature. This gives acid charge on ovendry-weight corncob of 0-2.58%. It was found that optimal pretreatment temperature is between 160 and 170 °C based on total xylose and glucose yields and thermal energy consumption in pretreatment. At 170 °C and acid charge of 2.2% on cob, total glucose yield and xylose recovery were 97% and 75%, respectively, which resulted in an overall monomeric sugar recovery of about 88%. Xylose concentration in the hydrolysate was about 12%, with xylose-to-acetic-acid ratio of 8 and to furan (furfural and hydroxymethylfurfural) of about 15.

Efficient fermentation of hemicellulosic sugars is critical for the bioconversion of lignocellulosics to ethanol. Efficient sugar uptake through the heterologous expression of yeast and fungal xylose/glucose transporters can improve fermentation if other metabolic steps are not rate limiting. Rectification of cofactor imbalances through heterologous expression of fungal xylose isomerase or modification of cofactor requirements in the yeast oxidoreductase pathway can reduce xylitol production while increasing ethanol yields, but these changes often occur at the expense of xylose utilization rates. Genetic engineering and evolutionary adaptation to increase glycolytic flux coupled with transcriptomic and proteomic studies have identified targets for further modification, as have genomic and metabolic engineering studies in native xylose fermenting yeasts.

As part of our study of the old (~2.5 Gyr) open cluster NGC 6819 in the Kepler field, we present photometric (Kepler and ground-based BVRcIc) and spectroscopic observations of the detached eclipsing binary WOCS 24009 (Auner 665; KIC 5023948) with a short orbital period of 3.6 days. WOCS 24009 is a triple-lined system, and we verify that the brightest star is physically orbiting the eclipsing binary using radial velocities and eclipse timing variations. The eclipsing binary components have masses M_B =1.090+/-0.010 Msun and M_C =1.075+/-0.013 Msun, and radii R_B =1.095+/-0.007 Rsun and R_C =1.057+/-0.008 Rsun. The bright non-eclipsing star resides at the cluster turnoff, and ultimately its mass will directly constrain the turnoff mass: our preliminary determination is M_A =1.251+/-0.057 Msun. A careful examination of the light curves indicates that the fainter star in the eclipsing binary undergoes a very brief period of total eclipse, which enables us to precisely decompose the light of the three stars and place them in the color-magnitude diagram. We also present improved analysis of two previously discussed detached eclipsing stars in NGC 6819 (WOCS 40007 and WOCS 23009) en route to a combined determination of the cluster's distance modulus (m-M)_V=12.38+/-0.04. Because this paper significantly increases the number of measured stars in the cluster, we can better constrain the age of the color-magnitude diagram to be 2.21+/-0.10+/-0.20 Gyr. Additionally, using all measured eclipsing binary star masses and radii, we constrain the age to 2.38+/-0.05+/-0.22 Gyr. The quoted uncertainties are estimates of measurement and systematic uncertainties (due to model physics differences and metal content), respectively., Comment: accepted to AJ, 13 figures, eclipse timings and radial velocity data will be available with the journal article

We describe here a useful metabolic engineering tool, multiple-gene-promoter shuffling (MGPS), to optimize expression levels for multiple genes. This method approaches an optimized gene overexpression level by fusing promoters of various strengths to genes of interest for a particular pathway. Selection of these promoters is based on the expression levels of the native genes under the same physiological conditions intended for the application. MGPS was implemented in a yeast xylose fermentation mixture by shuffling the promoters for GND2 and HXK2 with the genes for transaldolase ( TAL1 ), transketolase ( TKL1 ), and pyruvate kinase ( PYK1 ) in the Saccharomyces cerevisiae strain FPL-YSX3. This host strain has integrated xylose-metabolizing genes, including xylose reductase, xylitol dehydrogenase, and xylulose kinase. The optimal expression levels for TAL1 , TKL1 , and PYK1 were identified by analysis of volumetric ethanol production by transformed cells. We found the optimal combination for ethanol production to be GND2 - TAL1 - HXK2 - TKL1 - HXK2 - PYK1 . The MGPS method could easily be adapted for other eukaryotic and prokaryotic organisms to optimize expression of genes for industrial fermentation.

TASK-1 (KCNK3) and TASK-3 (KCNK9) are members of the two-pore domain potassium channel family and form either homomeric or heteromeric open-rectifier (leak) channels. Recent evidence suggests that these channels contribute to the resting potential and input resistance in several neuron types, including hippocampal CA1 pyramidal cells. However, the evidence for TWIK-related acid-sensitive potassium (TASK)-like conductances in inhibitory interneurons is less clear, and mRNA expression has suggested that TASK channels are expressed in only a subpopulation of interneurons. Here we use immunocytochemistry to demonstrate prominent TASK-3 protein expression in both parvalbumin-positive- and a subpopulation of glutamic acid decarboxylase (GAD)67-positive interneurons. In addition, a TASK-like current (modulated by both pH and bupivacaine) was detected in 30–50% of CA1 stratum oriens interneurons of various morphological classes. In most neurons, basic shifts in pH had a larger effect on the TASK-like current than acidic, suggesting that the current is mediated by TASK-1/TASK-3 heterodimers. These data suggest that TASK-like conductances are more prevalent in inhibitory interneurons than previously supposed.

Technologies for the production of alternative fuels are receiving increased attention owing to concerns over the rising cost of petrol and global warming. One such technology under development is the use of yeasts for the commercial fermentation of xylose to ethanol. Several approaches have been employed to engineer xylose metabolism. These involve modeling, flux analysis, and expression analysis followed by the targeted deletion or altered expression of key genes. Expression analysis is increasingly being used to target rate-limiting steps. Quantitative metabolic models have also proved extremely useful: they can be calculated from stoichiometric balances or inferred from the labeling of intermediate metabolites. The recent determination of the genome sequence for P. stipitis is important, as its genome characteristics and regulatory patterns could serve as guides for further development in this natural xylose-fermenting yeast or in engineered Saccharomyces cerevisiae. Lastly, strain selection through mutagenesis, adaptive evolution or from nature can also be employed to further improve activity.

Pichia stipitis is widely studied for its capacity to ferment d-xylose to ethanol. Strain improvement has been facilitated by recent completion of the P. stipitis genome. P. stipitis uses CUG to code for serine rather than leucine, as is the case for the universal genetic code thereby limiting the availability of heterologous drug resistance markers for transformation. Development of a modified selectable marker for resistance to bleomycin (Sh ble) and efficient excision of the marker after integration (loxP/Cre) should facilitate functional genomics and metabolic engineering in this yeast. The Sh ble marker did not code for an active protein in P. stipitis until four CUG codons were mutagenized to TTG, which is properly translated as leucine in yeasts that use the alternative yeast nuclear genetic code. The 18 CTG codons in Cre were mutagenized in a similar manner and the system was used to delete XYL2. The resulting xyl2Δ mutant did not use xylose as a carbon source.

The fermentation of xylose is essential for the bioconversion of lignocellulose to fuels and chemicals, but wild-type strains of Saccharomyces cerevisiae do not metabolize xylose, so researchers have engineered xylose metabolism in this yeast. Glucose transporters mediate xylose uptake, but no transporter specific for xylose has yet been identified. Over-expressing genes for aldose (xylose) reductase, xylitol dehydrogenase and moderate levels of xylulokinase enable xylose assimilation and fermentation, but a balanced supply of NAD(P) and NAD(P)H must be maintained to avoid xylitol production. Reducing production of NADPH by blocking the oxidative pentose phosphate cycle can reduce xylitol formation, but this occurs at the expense of xylose assimilation. Respiration is critical for growth on xylose by both native xylose-fermenting yeasts and recombinant S, cerevisiae. Anaerobic growth by recombinant mutants has been reported. Reducing the respiration capacity of xylose-metabolizing yeasts increases ethanol production. Recently, two routes for arabinose metabolism have been engineered in S. cerevisiae and adapted strains of Pichia stipitis have been shown to ferment hydrolysates with ethanol yields of 0.45 g g(-1) sugar consumed, so commercialization seems feasible for some applications.

The lack of industrially suitable microorganisms for converting biomass into fuel ethanol has traditionally been cited as a major technical roadblock to developing a bioethanol industry. In the last two decades, numerous microorganisms have been engineered to selectively produce ethanol. Lignocellulosic biomass contains complex carbohydrates that necessitate utilizing microorganisms capable of fermenting sugars not fermentable by brewers' yeast. The most significant of these is xylose. The greatest successes have been in the engineering of Gram-negative bacteria: Escherichia coli, Klebsiella oxytoca, and Zymomonas mobilis. E. coli and K. oxytoca are naturally able to use a wide spectrum of sugars, and work has concentrated on engineering these strains to selectively produce ethanol. Z. mobilis produces ethanol at high yields, but ferments only glucose and fructose. Work on this organism has concentrated on introducing pathways for the fermentation of arabinose and xylose. The history of constructing these strains and current progress in refining them are detailed in this review.

High-throughput screening requires simple assays that give reliable quantitative results. A microplate assay was developed for reducing sugar analysis that uses a 2,2′-bicinchoninic-based protein reagent. Endo-1,4-β-d-xylanase activity against oat spelt xylan was detected at activities of 0.002 to 0.011 IU ml−1. The assay is linear for sugar concentrations from 0 to 86 μg ml−1 and can also be used to assay protein concentrations (0 to 143 μg ml−1) on the same plate. A variety of temperatures and pH conditions can be used and, after incubation, the assay requires only one detection reagent and one heating step.

d -Xylulokinase (XK) is essential for the metabolism of d -xylose in yeasts. However, overexpression of genes for XK, such as the Pichia stipitis XYL3 gene and the Saccharomyces cerevisiae XKS gene, can inhibit growth of S . cerevisiae on xylose. We varied the copy number and promoter strength of XYL3 or XKS1 to see how XK activity can affect xylose metabolism in S . cerevisiae . The S . cerevisiae genetic background included single integrated copies of P . stipitis XYL1 and XYL2 driven by the S . cerevisiae TDH1 promoter. Multicopy and single-copy constructs with either XYL3 or XKS1 , likewise under control of the TDH1 promoter, or with the native P . stipitis promoter were introduced into the recombinant S . cerevisiae . In vitro enzymatic activity of XK increased with copy number and promoter strength. Overexpression of XYL3 and XKS1 inhibited growth on xylose but did not affect growth on glucose even though XK activities were three times higher in glucose-grown cells. Growth inhibition increased and ethanol yields from xylose decreased with increasing XK activity. Uncontrolled XK expression in recombinant S . cerevisiae is inhibitory in a manner analogous to the substrate-accelerated cell death observed with an S . cerevisiae tps1 mutant during glucose metabolism. To bypass this effect, we transformed cells with a tunable expression vector containing XYL3 under the control of its native promoter into the FPL-YS1020 strain and screened the transformants for growth on, and ethanol production from, xylose. The selected transformant had approximately four copies of XYL3 per haploid genome and had moderate XK activity. It converted xylose into ethanol efficiently.

The glucose analog 2-deoxyglucose (2-DOG) has been used to obtain mutants depressed for pentose metabolism. Some researchers have used 2-DOG alone whereas others have used it in the presence of a glucose-repressible carbon source. We examined both methods and screened mutant strains for improved use of xylose in the presence of glucose. Pichia stipitis mutants selected for growth on d-xylose in the presence of 2-DOG used xylose from a 1∶1 glucose:xylose mixture more rapidly than did their parents. One of these mutants, FPL-DX26, completely consumed xylose in the presence of glucose and produced 33g/L ethanol in 45h from 80 g/L of this sugar mixture. Mutants selected for growth on 2-DOG alone did not show significant improvement. Selection for growth on d-xylose in the presence of 2-DOG has been useful in developing parental strains for further genetic manipulation.

Transformation of Pichia stipitis is required to advance genetic studies and development of xylose metabolism in this yeast. To this end, we used P. stipitis URA3 (PsURA3) to disrupt P. stipitis LEU2 in a P. stipitis ura3 mutant. A highly fermentative P. stipitis mutant (FPL-DX26) was selected for resistance to 5'-fluoroorotic acid to obtain P. stipitis FPL-UC7 (ura3-3). A URA3:lacZ "pop-out" cassette was constructed containing PsURA3 flanked by direct repeats from segments of the lacZ reading frame. The P. stipitis LEU2 gene (PsLEU2) was cloned from a P. stipitis CBS 6054 genomic library through homology to Saccharomyces cerevisiae LEU2, and a disruption cassette was constructed by replacing the PsLEU2 reading sequence with the PsURA3:lacZ cassette. FPL-UC7 (ura3-3) was transformed with the disruption cassette, and a site-specific integrant was identified by selecting for the Leu- Ura+ phenotype. The ura3 marker was recovered from this strain by plating cells onto 5'-fluoroorotate and screening for spontaneous URA3 deletion mutants. Excision of the flanked PsURA3 gene resulted in the Leu- Ura- phenotype. The double auxotrophs are stable and can be transformed at a high frequency by PsLEU2 or PsURA3 carried on autonomous-replication-sequence-based plasmids.

In Pichia stipitis , fermentative and pyruvate decarboxylase (PDC) activities increase with diminished oxygen rather than in response to fermentable sugars. To better characterize PDC expression and regulation, two genes for PDC ( PsPDC1 and PsPDC2 ) were cloned and sequenced from P. stipitis CBS 6054. Aside from Saccharomyces cerevisiae , from which three PDC genes have been characterized, P. stipitis is the only organism from which multiple genes for PDC have been identified and characterized. PsPDC1 and PsPDC2 have diverged almost as far from one another as they have from the next most closely related known yeast gene. PsPDC1 contains an open reading frame of 1,791 nucleotides encoding 597 amino acids. PsPDC2 contains a reading frame of 1,710 nucleotides encoding 570 amino acids. An 81-nucleotide segment in the middle of the β domain of PsPDC1 codes for a unique segment of 27 amino acids, which may play a role in allosteric regulation. The 5′ regions of both P. stipitis genes include two putative TATA elements that make them similar to the PDC genes from S. cerevisiae , Kluyveromyces marxianus , and Hanseniaspora uvarum.

International audience; Forebrain circuits rely upon a relatively small but remarkably diverse population of GABAergic interneurons to bind and entrain large principal cell assemblies for network synchronization and rhythmogenesis. Despite the high degree of heterogeneity across cortical interneurons, members of a given subtype typically exhibit homogeneous developmental origins, neuromodulatory response profiles, morphological characteristics, neurochemical signatures and electrical features. Here we report a surprising divergence among hippocampal oriens-lacunosum moleculare (O-LM) projecting interneurons that have hitherto been considered a homogeneous cell population. Combined immunocytochemical, anatomical and electrophysiological interrogation of Htr3a-GFP and Nkx2-1-cre:RCE mice revealed that O-LM cells parse into a caudal ganglionic eminence-derived subpopulation expressing 5-HT3A receptors (5-HT(3A)Rs) and a medial ganglionic eminence-derived subpopulation lacking 5-HT(3A)Rs. These two cohorts differentially participate in network oscillations, with 5-HT3AR-containing O-LM cell recruitment dictated by serotonergic tone. Thus, members of a seemingly uniform interneuron population can exhibit unique circuit functions and neuromodulatory properties dictated by disparate developmental origins.

Anxiety disorders are common after stroke. However, information on how to treat them with psychotherapy in this population is highly limited. Modified cognitive-behaviour therapy (CBT) has the potential to assist. Two cases of individuals treated with modified CBT for anxiety after stroke are presented. The modification was required in light of deficits in executive and memory function in one individual and in the context of communication difficulties in the other. The anxiety symptoms were treated over seven and nine sessions, respectively. Both participants improved following the intervention, and these improvements were maintained at 3 month follow-ups. Further case-series and randomised controlled designs are required to support and develop modified CBT for those with anxiety after stroke. © 2013 © 2013 The Author(s). Published by Taylor & Francis.

Production of xylitol by Candida boidinii NRRL Y-17213 occurs under conditions of an oxygen limitation. The extent to which substrate is converted to xylitol and its coproducts (ethanol, other polyols, acetic acid), and the relative flow rates of substrate to energetic and biosynthetic pathways is controlled by the degree of oxygen limitation. With decrease in oxygen concentration in the inlet gas, for a constant dilution rate of 0.05 1/h. the specific oxygen uptake rate decreased from 1.30 to 0.36 mmol/gh Xylitol was not produced at specific oxygen uptake rates above 0.91 mmol/gh. Upon shift to lower oxygen rates, specific xylitol production rate increased more rapidly than specific ethanol production rate

Xylanases are classified into two major families (10 or F and 11 or G) of glycosyl hydrolases. Both use ion pair catalytic mechanisms and both retain anomeric configuration following hydrolysis. Family 10 xylanases are larger, more complex and produce smaller oligosaccharides; Family 11 xylanases are more specific for xylan. Alkaline-active and extreme-thermophilic enzymes are of particular interest. Such xylanases are being commercialized for bleaching pulps and other applications.

Two broad frameworks have informed much of the burgeoning literature on the World War II American homefront. One involves the impact of the warthe issue of World War II as a "watershed," a major divide in modern American history. The other concerns the nature of the war-the idea of World War II as the "Good War" of national unity, virtue, and success. Whether or not the Second World War was a watershed event has been

Giant reed was evaluated for enzymatic hydrolysis and simultaneous saccharification and fermentation (SSF) using a commercial cellulase/β-glucosidase and Scheffersomyces ( Pichia ) stipitis CBS 6054 for ethanol production following dilute-oxalic acid pretreatment. A response surface methodology with two input parameters – severity factor (SF) and oxalic acid concentration (OA) – was employed to optimize both enzymatic hydrolysis and SSF. Xylan content after dilute-OA pretreatment decreased with increasing SF and OA; almost complete hydrolysis was observed when the harsher pretreatment conditions were used. Glucan and lignin content showed an opposite trend with respect to xylan content after dilute-OA pretreatment. Accordingly, enzymatic hydrolysis and ethanol production reached 95% of glucan conversion and 18 g l −1 (75.3% of the maximum theoretical ethanol yield), respectively, with the pretreatment condition 4.05 SF and 5% OA w/w. Dilute-OA mediated pretreatment of giant reed followed by coupled saccharification and fermentation can be considered a promising methodology for second generation bioethanol production.

ABacillus sp (V1-4) was isolated from hardwood kraft pulp. It was capable of growing in diluted kraft black liquor at pH 11.5 and produced 49 IU (μmol xylose min−1 ml−1) of xylanase when cultivated in alkaline medium at pH 9. Maximal enzyme activity was obtained by cultivation in a defined alkaline medium with 2% birchwood xylan and 1% corn steep liquor at pH 9, but high enzyme production was also obtained on wheat bran. The apparent pH optimum of the enzyme varied with the pH used for cultivation and the buffer system employed for enzyme assay. With cultivation at pH 10 and assays performed in glycine buffer, maximal activity was observed at pH 8.5; with phosphate buffer, maximal activity was between pH 6 and 7. The xylanase temperature optimum (at pH 7.0) was 55°C. In the absence of substrate, at pH 9.0, the enzyme was stable at 50°C for at least 30 min. Elecrophoretic analysis of the crude preparation showed one predominant xylanase with an alkaline pl. Biobleaching studies showed that the enzyme would brighten both hardwood and softwood kraft pulp and release chromophores at pH 7 and 9. Because kraft pulps are alkaline, this enzyme could be used for prebleaching with minimal pH adjustment.

Microbial endoxylanases reduce chemical demand when they are applied to pulps prior to bleaching sequences. However, their mechanism of action is not fully understood, and the criteria for determining which enzymes are most effective are not well known. We examined the effects of two types of well-characterized endoxylanases from Streptomyces sp. TUB B-12-2 on the bleachability of kraft softwood and hardwood pulps. xyl 1a is a low molecular mass, high pl xylanase that forms mainly xylotriose but no xylose from oat spelts xylan. xyl 3 is a high molecular mass, low pl xylanase that forms mainly xylose and xylobiose as end products. xyl 3 released more reducing sugars than did xyl 1a, but both enzymes released about the same amount of total sugars. High performance liquid chromatography analysis of product sugars from pulp showed that at pH 7.0, xyl 1a formed oligosaccharides with a high degree of polymerization, but at pH 9.0 the action patterns of the two enzymes were similar. xyl 3 released 12% more chromophores from hardwood and 30% more from softwood pulp than did xyl 1a. Both enzymes enhanced brightness while reducing chemical demand, but xyl 3 was more effective. Synergism was noted in the prebleaching of softwood but not hardwood pulp. Both enzymes were active at pH 9, and the presence of pulp increased thermal stability.

We present the discovery of the totally eclipsing long-period (P = 771.8 d) binary system WOCS 23009 in the old open cluster NGC 6819 that contains both an evolved star near central hydrogen exhaustion and a low-mass (0.45 Msun) star. This system was previously known to be a single-lined spectroscopic binary, but the discovery of an eclipse near apastron using data from the Kepler space telescope makes it clear that the system has an inclination that is very close to 90 degrees. Although the secondary star has not been identified in spectra, the mass of the primary star can be constrained using other eclipsing binaries in the cluster. The combination of total eclipses and a mass constraint for the primary star allows us to determine a reliable mass for the secondary star and radii for both stars, and to constrain the cluster age. Unlike well-measured stars of similar mass in field binaries, the low-mass secondary is not significantly inflated in radius compared to model predictions. The primary star characteristics, in combination with cluster photometry and masses from other cluster binaries, indicates a best age of 2.62+/-0.25 Gyr, although stellar model physics may introduce systematic uncertainties at the ~10% level. We find preliminary evidence that the asteroseismic predictions for red giant masses in this cluster are systematically too high by as much as 8%., 38 pages,11 figures, accepted for ApJ

Fermentation of cellulosic and hemicellulosic sugars from biomass could resolve food-versus-fuel conflicts inherent in the bioconversion of grains. However, the inability to coferment glucose and xylose is a major challenge to the economical use of lignocellulose as a feedstock. Simultaneous cofermentation of glucose, xylose, and cellobiose is problematic for most microbes because glucose represses utilization of the other saccharides. Surprisingly, the ascomycetous, beetle-associated yeast Spathaspora passalidarum , which ferments xylose and cellobiose natively, can also coferment these two sugars in the presence of 30 g/liter glucose. S. passalidarum simultaneously assimilates glucose and xylose aerobically, it simultaneously coferments glucose, cellobiose, and xylose with an ethanol yield of 0.42 g/g, and it has a specific ethanol production rate on xylose more than 3 times that of the corresponding rate on glucose. Moreover, an adapted strain of S. passalidarum produced 39 g/liter ethanol with a yield of 0.37 g/g sugars from a hardwood hydrolysate. Metabolome analysis of S. passalidarum before onset and during the fermentations of glucose and xylose showed that the flux of glycolytic intermediates is significantly higher on xylose than on glucose. The high affinity of its xylose reductase activities for NADH and xylose combined with allosteric activation of glycolysis probably accounts in part for its unusual capacities. These features make S. passalidarum very attractive for studying regulatory mechanisms enabling bioconversion of lignocellulosic materials by yeasts.

Xylose utilization is essential for the efficient conversion of lignocellulose to ethanol. The objective of this review is to trace the development of xylose-fermenting yeast strains from their discovery in 1980. following initial reports, screens of known yeast identified five species of interest: Candida shehatae, Candida tenuis, Pachysolen tannophilus, Pichia segobiensis , and Pichia stipitis, Candida shehatae strains can be deivided into three varieties. Pachysolen tannophilus and Pichia stipilis have been studied most extensively and have the best-understood genetic systems. Improved mutants of P. tannophilis have been obtained by selecting for an inability to oxidize ethanol ( eth ) and for rapid growth on xylitol and nitrate,. Improved P. stipitis mutants have been obtained by selecting for flocculation decreased utilization of glucose,a nd growth on noninductive carbon sources. Bacterial xylose isomerase has been cloned and expressed in S. cerevisiae and Schizosaccharomyces pombe , but he heterologous enzyme is inactive. Xylose reductase and xylitol dehydrogenase have been cloned from P. stipitis and expressed in Saccharomyces cerevisiae , giveing rise to transformant S. cerevisiae that grow on oxylose but that ferment it poorly. A transformation and expression system based on the URA3 marker has recently been developed for P. stipilitis so that contemporary genetic methods may be brough to bear on this organism.

Of the three primary phylogenetic domains--Archaea (archaebacteria), Bacteria (eubacteria), and Eucarya (eukaryotes)--Archaea is the least understood in terms of its diversity, physiologies, and ecological panorama. Although many species of Crenarchaeota (one of the two recognized archaeal kingdoms sensu Woese [Woese, C. R., Kandler, O. & Wheelis, M. L. (1990) Proc. Natl. Acad. Sci. USA 87, 4576-4579]) have been isolated, they constitute a relatively tight-knit cluster of lineages in phylogenetic analyses of rRNA sequences. It seemed possible that this limited diversity is merely apparent and reflects only a failure to culture organisms, not their absence. We report here phylogenetic characterization of many archaeal small subunit rRNA gene sequences obtained by polymerase chain reaction amplification of mixed population DNA extracted directly from sediment of a hot spring in Yellowstone National Park. This approach obviates the need for cultivation to identify organisms. The analyses document the existence not only of species belonging to well-characterized crenarchaeal genera or families but also of crenarchaeal species for which no close relatives have so far been found. The large number of distinct archaeal sequence types retrieved from this single hot spring was unexpected and demonstrates that Crenarchaeota is a much more diverse group than was previously suspected. The results have impact on our concepts of the phylogenetic organization of Archaea.

Pichia stipitis CBS 6054 ferments D-Xylose to ethanol in a medium containing corn steep liquor as the only source of nitrogen, amino acids, vitamins and other nutrients. The ethanol yield and fermentation rate compare favorably to those obtained with media containing more expensive sources of nitrogen, vitamins and amino acids. Corn steep liquor is a good source of nutrients that can support growth and fermentation activity of this xylose fermenting yeast.

Industrial biotechnology aims to develop robust microbial cell factories, such as Saccharomyces cerevisiae, to produce an array of added value chemicals presently dominated by petrochemical processes. Xylose is the second most abundant monosaccharide after glucose and the most prevalent pentose sugar found in lignocelluloses. Significant research efforts have focused on the metabolic engineering of S similar to cerevisiae for fast and efficient xylose utilization. This study aims to metabolically engineer S similar to cerevisiae, such that it can consume xylose as the exclusive substrate while maximizing carbon flux to biomass production. Such a platform may then be enhanced with complementary metabolic engineering strategies that couple biomass production with high value-added chemical. Saccharomyces cerevisiae, expressing xylose reductase, xylitol dehydrogenase and xylulose kinase, from the native xylose-metabolizing yeast Pichia stipitis, was constructed, followed by a directed evolution strategy to improve xylose utilization rates. The resulting S similar to cerevisiae strain was capable of rapid growth and fast xylose consumption producing only biomass and negligible amount of byproducts. Transcriptional profiling of this strain was employed to further elucidate the observed physiology confirms a strongly up-regulated glyoxylate pathway enabling respiratory metabolism. The resulting strain is a desirable platform for the industrial production of biomass-related products using xylose as a sole carbon source.

Although vastly outnumbered, inhibitory interneurons critically pace and synchronize excitatory principal cell populations to coordinate cortical information processing. Precision in this control relies upon a remarkable diversity of interneurons primarily determined during embryogenesis by genetic restriction of neuronal potential at the progenitor stage. Like their neocortical counterparts, hippocampal interneurons arise from medial and caudal ganglionic eminence (MGE and CGE) precursors. However, while studies of the early specification of neocortical interneurons are rapidly advancing, similar lineage analyses of hippocampal interneurons have lagged. A "hippocampocentric" investigation is necessary as several hippocampal interneuron subtypes remain poorly represented in the neocortical literature. Thus, we investigated the spatiotemporal origins of hippocampal interneurons using transgenic mice that specifically report MGE- and CGE-derived interneurons either constitutively or inducibly. We found that hippocampal interneurons are produced in two neurogenic waves between E9-E12 and E12-E16 from MGE and CGE, respectively, and invade the hippocampus by E14. In the mature hippocampus, CGE-derived interneurons primarily localize to superficial layers in strata lacunosum moleculare and deep radiatum, while MGE-derived interneurons readily populate all layers with preference for strata pyramidale and oriens. Combined molecular, anatomical, and electrophysiological interrogation of MGE/CGE-derived interneurons revealed that MGE produces parvalbumin-, somatostatin-, and nitric oxide synthase-expressing interneurons including fast-spiking basket, bistratified, axo-axonic, oriens-lacunosum moleculare, neurogliaform, and ivy cells. In contrast, CGE-derived interneurons contain cholecystokinin, calretinin, vasoactive intestinal peptide, and reelin including non-fast-spiking basket, Schaffer collateral-associated, mossy fiber-associated, trilaminar, and additional neurogliaform cells. Our findings provide a basic blueprint of the developmental origins of hippocampal interneuron diversity.

Cellulosic biomass is an abundant and underused substrate for biofuel production. The inability of many microbes to metabolize the pentose sugars abundant within hemicellulose creates specific challenges for microbial biofuel production from cellulosic material. Although engineered strains of Saccharomyces cerevisiae can use the pentose xylose, the fermentative capacity pales in comparison with glucose, limiting the economic feasibility of industrial fermentations. To better understand xylose utilization for subsequent microbial engineering, we sequenced the genomes of two xylose-fermenting, beetle-associated fungi, Spathaspora passalidarum and Candida tenuis . To identify genes involved in xylose metabolism, we applied a comparative genomic approach across 14 Ascomycete genomes, mapping phenotypes and genotypes onto the fungal phylogeny, and measured genomic expression across five Hemiascomycete species with different xylose-consumption phenotypes. This approach implicated many genes and processes involved in xylose assimilation. Several of these genes significantly improved xylose utilization when engineered into S. cerevisiae , demonstrating the power of comparative methods in rapidly identifying genes for biomass conversion while reflecting on fungal ecology.