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Endophenotypes for alcohol use disorder are well known and may reflect paternal exposure effects passed down to offspring via epigenetic mechanisms. Previously, we showed that paternal alcohol exposure prior to conception attenuates the acquisition of operant alcohol self-administration. We now test whether paternal alcohol exposure alters their offsprings' behavioral responses to alcohol (endophenotypes) and global DNA methylation levels in reward-related brain regions.Adult male rats were exposed to alcohol vapors or air for 6 weeks and mated with alcohol-naïve females 8 weeks later. Adult male and female offspring of the alcohol- and control-sired litters were tested on three behaviors 30 m after gavage with water or alcohol (1.5 g/kg): open field, elevated plus maze, and accelerating rotarod. Global DNA methylation levels in sperm, nucleus accumbens, and prefrontal cortex were examined in male sires and in another group of offspring.Alcohol-sired males showed less anxiety-like behavior in the elevated plus maze that was not affected by alcohol administration. By contrast, alcohol had anxiolytic effects in the open field in male offspring only with no paternal alcohol effect. Neither paternal alcohol exposure nor alcohol administration altered locomotor activity in either sex. Sex-specific effects of paternal alcohol exposure were seen in the rotarod test. Alcohol-sired male offspring showed blunted sensitivity to the alcohol's motor-impairing effects, whereas alcohol-sired female offspring showed enhanced sensitivity. Global DNA methylation was altered in the sperm of alcohol-exposed males, but no changes were seen in their offspring.Paternal alcohol exposure prior to conception has sex- and task-dependent effects on unconditioned behaviors in their offspring.

Amino acids and related targets are typically produced by well-characterized heterotrophs including Corynebacterium glutamicum and Escherichia coli. Cyanobacteria offer an opportunity to supplant these sugar-intensive processes by instead directly utilizing atmospheric CO2 and sunlight. Synechococcus elongatus UTEX 2973 (hereafter UTEX 2973) is a particularly promising photoautotrophic platform due to its fast growth rate. Here, we first engineered UTEX 2973 to overproduce l-lysine (hereafter lysine), after which both cadaverine and glutarate production were achieved through further pathway engineering. To facilitate metabolic engineering, the relative activities of a subset of previously uncharacterized promoters were investigated, in each case, while also comparing the effects of both chromosomal (from neutral site NS3) and episomal (from pAM4788) expressions. Using these parts, lysine overproduction in UTEX 2973 was engineered by introducing a feedback-resistant copy of aspartate kinase (encoded by lysCfbr) and a lysine exporter (encoded by ybjE), both from E. coli. While chromosomal expression resulted in lysine production up to just 325.3 ± 14.8 mg/L after 120 h, this was then increased to 556.3 ± 62.3 mg/L via plasmid-based expression, also surpassing prior reports of photoautotrophic lysine bioproduction. Lastly, additional products of interest were then targeted by modularly extending the lysine pathway to glutarate and cadaverine, two 5-carbon, bioplastic monomers. By this approach, glutarate has so far been produced at final titers reaching 67.5 ± 2.2 mg/L by 96 h, whereas cadaverine has been produced at up to 55.3 ± 6.7 mg/L. Overcoming pathway and/or transport bottlenecks, meanwhile, will be important to improving upon these initial outputs.

A novel dissolution method that allows for the total solvation of high-concentration, high-molecular-weight polyaniline (PANi) doped with (+)-camphor-10-sulfonic acid (CSA) is reported. Preparation of 12-16 wt % 65,000 Da PANi solutions in N,N-dimethylformamide is achievable using a simple one-pot method. Doped polyaniline solutions in common organic solvents were processed into nanofibers using a convenient single-nozzle electrospinning technique. The electrospinning of PANi-CSA into nanofibrous membranes generated substrates that were subsequently employed in colorimetric gas sensing. These substrates demonstrated linearity of resistivity upon exposure to 50-5500 ppm ammonia at ambient (50 ± 10% RH) and high (80% RH) humidity.

Microbial production of biopolymers represents a promising, sustainable alternative to current approaches for plastic production. Cyanophycin synthetase 1 (CphA1) produces cyanophycin - an attractive biopolymer consisting of a poly-L-aspartic acid backbone decorated with L-arginine side groups. In this work, a series of CphA1 enzymes from different bacteria were screened for heterologous cyanophycin production in engineeredEscherichia coli, from which it was found that CphA1 fromTatumella morbirosei(TmCphA1) was especially productive.TmCphA1 was capable of supporting up to ~2-fold greater yields of insoluble cyanophycin than any other tested CphA1 enzymes, including 10.8-times more than CphA1 fromSynechocystissp. PCC6308. Finally, using a bench-scale bioreactor, cyanophycin production byTmCphA1-expressingE. colireached up to 1.9 g per liter of culture by 48 h.

Carbon loss in the form of CO2 is an intrinsic and persistent challenge faced during conventional and advanced biofuel production from biomass feedstocks. Current mechanisms for increasing carbon conservation typically require the provision of reduced co-substrates as additional reducing equivalents. This need can be circumvented, however, by exploiting the natural heterogeneity of lignocellulosic sugars mixtures and strategically using specific fractions to drive complementary CO2 emitting vs. CO2 fixing pathways. As a demonstration of concept, a coculture-coproduction system was developed by pairing two catabolically orthogonal Escherichia coli strains; one converting glucose to ethanol (G2E) and the other xylose to succinate (X2S). 13C-labeling studies reveled that G2E + X2S cocultures were capable of recycling 24% of all evolved CO2 and achieved a carbon conservation efficiency of 77%; significantly higher than the 64% achieved when all sugars are instead converted to just ethanol. In addition to CO2 exchange, the latent exchange of pyruvate between strains was discovered, along with significant carbon rearrangement within X2S.

Here we describe a universal approach for plasmid-free genome engineering in cyanobacteria that exploits the polyploidy of their chromosomes as a natural counterselection system. Rather than being delivered via replicating plasmids, genes encoding for DNA modifying enzymes are instead integrated into essential genes on the chromosome by allelic exchange, as facilitated by antibiotic selection, a process that occurs readily and with only minor fitness defects. By virtue of the essentiality of these integration sites, full segregation is never achieved, with the strain instead remaining as a merodiploid so long as antibiotic selection is maintained. As a result, once the desired genome modification is complete, removal of antibiotic selection results in the gene encoding for the DNA modifying enzyme to then be promptly eliminated from the population. Proof of concept of this new and generalizable strategy is provided using two different site-specific recombination systems, CRE-lox and DRE-rox, in the fast-growing cyanobacterium Synechococcus sp. PCC 7002, as well as CRE-lox in the model cyanobacterium Synechocystis sp. PCC 6803. Reusability of the method, meanwhile, is demonstrated by constructing a high-CO2 requiring and markerless Δndh3 Δndh4 ΔbicA ΔsbtA mutant of Synechococcus sp. PCC 7002. Overall, this method enables the simple and efficient construction of stable and unmarked mutants in cyanobacteria without the need to develop additional shuttle vectors nor counterselection systems.

Drug addiction is a serious relapsing disease that has high costs to society and to the individual addicts. Treatment of these addictions is still in its nascency, with only a few examples of successful therapies. Therapeutic response depends upon genetic, biological, social, and environmental components. A role for genetic makeup in the response to treatment has been shown for several addiction pharmacotherapies with response to treatment based on individual genetic makeup. In this chapter, we will discuss the role of genetics in pharmacotherapies, specifically for cocaine, alcohol, and opioid dependences. The continued elucidation of the role of genetics should aid in the development of new treatments and increase the efficacy of existing treatments.

In 2017, a single adult Triatoma sanguisuga (LeConte) (Hemiptera: Reduviidae) was identified from Indian Cave State Park in Nemaha County, NE. The following summer, a single adult specimen was found in a 4-H insect collection at the Nebraska State Fair from Richardson County. A subsequent investigation found that these were collected from a residence in Richardson County and three more adults were collected in 2019. In 2020, the latter three kissing bugs were submitted to Public Health Command-Central’s DoD Food Analysis and Diagnostic Laboratory (DOD FADL) for confirmatory species identification and diagnostic testing for Trypanosoma cruzi, the causative agent of Chagas disease. One specimen tested positive for T. cruzi using a real-time dual-target PCR screen followed by confirmatory dual-target traditional PCR. Based on these findings, a survey plan was developed for Richardson and surrounding counties in southeast Nebraska. In July of 2020, two adult and seven nymphs (multiple instars) of T. sanguisuga were collected in Richardson County, one of which tested positive for T. cruzi. This is the first record of an established population of T. sanguisuga and T. cruzi-infected kissing bugs in Nebraska.

Robust systematic approaches for the metabolic engineering of cell factories remain elusive. The available models for predicting phenotypical responses and mechanisms are incomplete, particularly within the context of compound toxicity that can be a significant impediment to achieving high yields of a target product. This study describes a Multi-Omic Based Production Strain Improvement (MOBpsi) strategy that is distinguished by integrated time-resolved systems analyses of fed-batch fermentations. As a case study, MOBpsi was applied to improve the performance of an Escherichia coli cell factory producing the commodity chemical styrene. Styrene can be bio-manufactured from phenylalanine via an engineered pathway comprised of the enzymes phenylalanine ammonia lyase and ferulic acid decarboxylase. The toxicity, hydrophobicity, and volatility of styrene combine to make bio-production challenging. Previous attempts to create styrene tolerant E. coli strains by targeted genetic interventions have met with modest success. Application of MOBpsi identified new potential targets for improving performance, resulting in two host strains (E. coli NST74ΔaaeA and NST74ΔaaeA cpxPo) with increased styrene production. The best performing re-engineered chassis, NST74ΔaaeA cpxPo, produced ∼3 × more styrene and exhibited increased viability in fed-batch fermentations. Thus, this case study demonstrates the utility of MOBpsi as a systematic tool for improving the bio-manufacturing of toxic chemicals.

Over the last decade, the northeast Pacific experienced marine heatwaves that caused devastating marine ecological impacts with socioeconomic implications. Here we use two different attribution methods and show that forcing by elevated greenhouse gases levels has virtually certainly caused the multi-year persistent 2019–2021 marine heatwave. There is less than 1% chance that the 2019–2021 event with ~3 years duration and 1.6 ∘C intensity could have happened in the absence of greenhouse gases forcing. We further discover that the recent marine heatwaves are co-located with a systematically-forced outstanding warming pool, which we attribute to forcing by elevated greenhouse gases levels and the recent industrial aerosol-load decrease. The here-detected Pacific long-term warming pool is associated with a strengthening ridge of high-pressure system, which has recently emerged from the natural variability of climate system, indicating that they will provide favorable conditions over the northeast Pacific for even more severe marine heatwave events in the future.

Residue removal from maize (Zea mays) fields offers an opportunity to increase farmer profits, but potential tradeoffs for water dynamics and crop performance merit further evaluation. This study, established in 2014, compared the effects of two tillage practices (no-till and conventional) and two residue management practices (harvested vs. kept in place) on maize grain yields, water infiltration, evapotranspiration, and soil physical attributes. On average, maize grain yields under limited irrigation increased with residue retention by 1.1 Mg ha−1 year−1 between 2016 and 2018, but tillage had no significant effect. Total infiltration (over 30 min) was higher with residue retention. Neither tillage nor residue management had a significant impact on evapotranspiration during the vegetative growth stage. However, there was a significant residue by tillage interaction where vegetative evapotranspiration was reduced by no-till and residue retention. Conversely, penetrometer resistance was significantly reduced by both tillage and residue retention. Volumetric water content in the soil profile at planting was higher with residue retention. These results suggest that plots with residue removal would on average require 60 mm year−1 of additional irrigation to attain the same yields as fields with residue retention. In summary, our findings suggest that high rates of crop residue removal under limited irrigation in a semiarid environment can negatively affect water conservation and yields, and that tradeoffs surrounding residue export need to be fully considered in land management and policy decisions.

Over the last decade, the northeast Pacific experienced strong marine heatwaves (MHWs) that produced devastating marine ecological impacts and received major societal concerns. Here, we assess the link between the well-mixed greenhouse gas (GHG) forcing and the occurrence probabilities of the duration and intensity of the North Pacific MHWs. We investigate whether GHG forcing was necessary for the North Pacific MHWs to occur and whether it is a sufficient cause for such events to continue to repeatedly occur in the 21st Century. To begin with, we apply attribution technique on the long-term SST time series, and detect a region of systematically and externally-forced SST increase -- the long-term warming pool -- co-located with the past notably Blob-like SST anomalies. We further show that the anthropogenic signal has recently emerged from the natural variability of SST over the warming pool, which we attribute primarily to increased GHG concentrations, with anthropogenic aerosols playing a secondary role.After we demonstrate that the GHG forcing has a dominant influence on the base climate state in the region, we pursue an event attribution analysis for MHWs on a more localized region. Extreme event attribution analysis reveals that GHG forcing is a necessary, but not sufficient, causation for the multi-year persistent MHW events in the current climate, such as that happened in 2014/2015 and 2019/2020. However, the occurrence of the 2019/2020 (2014/2015) MHW was extremely unlikely in the absence of GHG forcing. Thus, as GHG emissions continue to firmly rise, it is very likely that GHG forcings will become a sufficient cause for events of the magnitude of the 2019/2020 record event.

Over the last decade, the northeast Pacific (NP) experienced strong marine heatwaves (MHWs) that produced devastating marine ecological impacts and received major societal concerns. Here, we assess the link between the well-mixed greenhouse gas (GHG) forcing and the occurrence probabilities of the duration and intensity of the NP MHWs. To begin with, we apply attribution technique on the SST time series, and detect a region of systematically and externally-forced SST increase -- the long-term warming pool -- co-located with the past notably Blob-like SST anomalies. The anthropogenic signal has recently emerged from the natural variability of SST over the warming pool, which we attribute primarily to increased GHG concentrations, with anthropogenic aerosols playing a secondary role. With extreme event attribution technique, we further show that GHG forcing is a necessary, but not a sufficient, causation for the multi-year persistent MHW events in the current climate, such as that happened in 2019/2020 over the warming pool. However, the occurrence of the 2019/2020 MHW was extremely unlikely in the absence of GHG forcing. Thus, as GHG emissions continue to firmly rise, it is very likely that GHG forcings will become a sufficient cause for events of the magnitude of the 2019/2020 record event.

In the Pacific Ocean, off-equatorial Rossby waves (RWs), initiated by atmosphere-ocean interaction, modulate the inter-annual variability of the thermocline. In this study, we explore the resulting potential gain in predictability of central tropical Pacific primary production, which in this region strongly depends on the supply of macronutrients from below the thermocline. We use a decadal prediction system based on the Max Planck Institute Earth system model to demonstrate that for the time period 1998–2014 properly initialized RWs explain an increase in predictability of net primary productivity (NPP) in the off-equatorial central tropical Pacific. We show that, for up to 5 years in advance, predictability of NPP derived from the decadal prediction system is significantly larger than that derived from persistence alone, or an uninitialized historical simulation. The predicted signal can be explained by the following mechanism: off-equatorial RWs are initiated in the eastern Pacific and travel towards the central tropical Pacific on a time scale of 2–6 years. On their arrival the RWs modify the depths of both thermocline and nutricline, which is fundamental to the availability of nutrients in the euphotic layer. Local upwelling transports nutrients from below the nutricline into the euphotic zone, effectively transferring the RW signal to the near-surface ocean. While we show that skillful prediction of central off-equatorial tropical Pacific NPP is possible, we open the door for establishing predictive systems for food web and ecosystem services in that region.

Amino acids and related targets are typically produced by well-characterized heterotrophs including

Background Brain imaging and genetics are fields acquiring data at increasing speed, but more information does not always result in a better understanding of the underlying biology. We developed the ProcessGeneLists (PGL) approach to use genetics and mRNA gene expression data to generate regions of interest for imaging studies. Methods We applied PGL to past suicide attempt (ATT): We averaged the mRNA expression levels of genes (n = 130) possibly associated with ATT (p ≤ 10−3 in a published genome-wide association study, GWAS) in each brain region studied in the Human Allen Brain Atlas (6 ex-vivo brains, 158 to 946 regions/brain have mRNA expression data) and compared that to the averaged mRNA expression levels of all other genes in each region in each brain in the atlas. Results PGL revealed 8 regions where “attempt-related genes” were differentially expressed (Wilcoxon test with Bonferroni correction 8.88−11 = −6) and dorsolateral prefrontal cortex (dlPFC pFWE Conclusion PGL uncovered a brain function/genotype interaction in ATT by using published GWAS data to inform imaging studies. This could inform individualized therapies in the future.

Phenol is an important petrochemical that is conventionally used as a precursor for synthesizing an array of plastics and fine chemicals. As an emerging alternative to its traditional petrochemical production, multiple enzyme pathways have been engineered to date to enable its renewable biosynthesis from biomass feedstocks, each incorporating unique enzyme chemistries and intermediate molecules. Leveraging all three of the unique phenol biosynthesis pathways reported to date, a series of synthetic ‘metabolic funnels’ was engineered, each with the goal of maximizing net precursor assimilation and flux towards phenol via the parallel co-expression of multiple distinct pathways within the same Escherichia coli host. By constructing and evaluating all possible binary and tertiary pathway combinations, one ‘funnel’ was ultimately identified, which supported enhanced phenol production relative to all three individual pathways by 16 to 69%. Further host engineering to increase endogenous precursor availability then allowed for 26% greater phenol production, reaching a final titer of 554 ± 19 mg/L and 28.8 ± 0.34 mg/g yield on glucose. Lastly, using a diphasic culture including dibutyl phthalate for in situ phenol extraction, final titers were further increased to a maximum of 812 ± 145 mg/L at a yield of 40.6 ± 7.2 mg/g. The demonstrated ‘funneling’ pathway holds similar promise in support of phenol production by other, non-E. coli hosts, while this general approach can be readily extended towards a diversity of other value-added bioproducts of interest.

Background: The α1 antagonist doxazosin reduces cocaine use in individuals with cocaine use disorder (CUD) through a functional polymorphism of the α1 adrenoreceptor. The regulatory role of the α1 ...

Pediatric patients are often sedated for magnetic resonance imaging (MRI) scans to ensure images are of diagnostic quality. However, access time for MRIs requiring sedation is often long due to high patient volumes and limited sedation resources. This study examined the effectiveness of an MRI Try Without sedation program to decrease the wait time for obtaining an MRI while simultaneously ensuring diagnostic-quality images. A retrospective chart review was performed on subjects who utilized the MRI Try Without program from April 2014 through June 2015 at a dedicated pediatric institution. Child life specialist preparations and access time (i.e. time from exam ordered to exam completed) were recorded in each patient’s electronic medical record. MRI images were evaluated for image quality by a pediatric neuroradiologist. A total of 134 patients participated in the MRI Try Without program (mean age: 6.9±1.7 years), all of whom received interventions from a child life specialist. The average number of days between when the order was placed and when the MRI was completed using the MRI Try Without program was 15.4±18.5 days, while the third-available appointment for sedation/anesthesia was 46.2 days (standard deviation [SD]±15.7 days). Nearly all patients received a “good” or “very good” image quality determination (87.3%) and only 5 (3.8%) patients were recommended for repeat examination for diagnostic-quality images. Utilization of an MRI Try Without sedation program, with child life specialist interventions, decreased the wait time for obtaining an MRI while still providing diagnostic-quality images.

Microbial biosynthesis of fuels and chemicals represents a promising route for their renewable production. Product toxicity, however, represents a common challenge limiting the efficacy of this approach. Integrated bioprocesses incorporating in situ product separation are poised to help address this intrinsic problem, but suffer their own unique shortcomings. To improve and expand the utility of this versatile bioprocessing strategy, recent innovations have focused on developing more effective separation materials and novel process configurations, as well as adapting designs to accommodate semi-continuous modes of operation. As a result, integrated bioprocesses are finding new applications to aid the biosynthesis of an ever-growing list of bioproducts. Emerging applications, meanwhile, are exploring the further expansion of such designs to interface microbial and chemical catalysts, leading to new and versatile routes for the one-pot synthesis of an even greater diversity of renewable products.

Fermentation of lignocellulosic sugar mixtures is often suboptimal due to inefficient xylose catabolism and sequential sugar utilization caused by carbon catabolite repression. Unlike in conventional applications employing a single engineered strain, the alternative development of synthetic microbial communities facilitates the execution of complex metabolic tasks by exploiting the unique community features, including modularity, division of labor, and facile tunability. A series of synthetic, catabolically orthogonal coculture systems were systematically engineered, as derived from either wild-type Escherichia coli W or ethanologenic LY180. Net catabolic activities were effectively balanced by simple tuning of the inoculum ratio between specialist strains, which enabled coutilization (98% of 100 g L-1 total sugars) of glucose-xylose mixtures (2:1 by mass) for both culture systems in simple batch fermentations. The engineered ethanologenic cocultures achieved ethanol titer (46 g L-1), productivity (488 mg L-1 h-1), and yield (∼90% of theoretical maximum), which were all significantly increased compared to LY180 monocultures.

OBJECTIVE: The use of synthetic cannabinoid (SC) products has become popular in recent years, but data regarding their impact on hospital stays are limited. The impact of SC and cannabis use on hospital length of stay and doses of antipsychotics at discharge was assessed in this study. METHOD: The sample consisted of inpatients with discharge diagnoses of bipolar disorder, schizophrenia, or other psychotic disorders. Medical records of patients with self-reported SC use and negative urine drug screens (SC group, n = 77), with cannabis use confirmed by urine drug screen (cannabis group, n = 248), and with no drug use confirmed by urine drug screen (no-drug group, n = 1,336) were examined retrospectively. RESULTS: Length of stay (mean [SD] days) significantly differed (p < .001) among the SC (8.29 [4.29]), cannabis (8.02 [5.21]), and no-drug groups (10.19 [9.08]). Antipsychotic doses (chlorpromazine milligram equivalent doses) also significantly differed (p = .002) among the SC (254.64 [253.63]), cannabis (219.16 [216.71]), and no-drug groups (294.79 [287.85]). Unadjusted and adjusted pairwise comparisons showed that the cannabis group had a shorter length of stay (p < .001) and received lower doses of antipsychotics (p = .003) than the no-drug group. SC users did not differ significantly from the other two groups in either length of stay or doses of antipsychotics. CONCLUSIONS: Our findings suggest that acute SC exposure is not predictive of a more prolonged time for response to antipsychotic medications or of a need for larger doses of these medications compared with cannabis users.

Arctic coastal erosion damages infrastructure, threatens coastal communities, and releases organic carbon from permafrost. However, the magnitude, timing and sensitivity of coastal erosion increase to global warming remain unknown. Here, we project the Arctic-mean erosion rate to roughly double by 2100 and very likely exceed its historical range of variability by mid-21st century. The sensitivity of erosion to warming also doubles, reaching 0.4-0.5 m year-1 oC-1 and 2.3-2.8 TgC year-1 oC-1 by the end of the century under moderate and high-emission scenarios. Our first 21st-century pan-Arctic coastal erosion rate projections should inform policy makers on coastal conservation and socioeconomic planning. Our organic carbon flux projections also lay out the path for future work to investigate the impact of Arctic coastal erosion on the changing Arctic Ocean, on its role as a global carbon sink, and on the permafrost-carbon feedback.

Familial transmission of alcohol use disorder reflects genetic and environmental factors. Paternal alcohol exposure may affect rodent offspring via epigenetic modifications transmitted through the male germ line. While such exposure alters alcohol sensitivity in mouse offspring, no studies examined if it impacts the development of operant alcohol self-administration in rats. We exposed male (sires) Wistar rats to chronic intermittent ethanol in vapour chambers (16 h/day; 5 days/week) or to air for 6 weeks. Eight weeks later, rats were mated with alcohol-naive females. Adult alcohol- and control-sired F1 offspring were assessed in acquisition of alcohol self-administration in which increasing alcohol concentrations (2.5%, 5% and 10%, v/v) were delivered after one lever press (fixed ratio 1 or FR1). Prior to alcohol sessions, rats were trained to lever press for food delivery under an FR1 schedule of reinforcement. DNA methylation levels of the brain derived neurotrophic factor (Bdnf) gene were measured in sperm, nucleus accumbens (NAc) and medial prefrontal cortex (mPFC) in sires and in offspring. Alcohol-exposed sires had lower Bdnf DNA methylation levels in NAc and greater methylation levels in mPFC. Although this pattern was not recapitulated in offspring, alcohol-sired offspring of both sexes did show aberrant Bdnf DNA methylation patterns compared to control-sired offspring. Alcohol-sired offspring self-administered less alcohol (5% and 10%) with no group differences in food responding. Results indicate that paternal alcohol exposure prior to conception protects against alcohol's initial reinforcing effects but the pattern of dysregulated Bdnf methylation in reward-related circuitry did not mimic changes seen in sires.

OBJECTIVE: Prior research demonstrated that the δ-opioid receptor (OPRD1) rs678849 variant influences opioid use in African Americans treated with methadone. We examined whether this variant moderated cocaine and opioid use in our clinical cohort of methadone and disulfiram treated recipients. METHODS: Cocaine and opioid co-dependent patients were stabilized for two weeks on methadone and subsequently randomized into groups treated with either methadone+placebo (n = 37) or methadone+disulfiram (250 mg/day, n = 33) for 12 weeks. RESULTS: A drop in cocaine positive urines was found in the OPRD1 CC genotype group compared to T-allele carrier patients treated with methadone+disulfiram (P < 0.0001), but not in the methadone+placebo group. No difference in opioid positive urines were found among each genotype group in either treatment group. CONCLUSION: These findings suggested that rs678849 genotype may predict treatment response of disulfiram for cocaine use in patients with co-occurring opioid and cocaine dependence.

The superfamily Trombiculoidea is a large and diverse group of acarines that comprises six families; of these, the families Trombiculidae and Leeuwenhoekiidae are characterized by larvae commonly known as chiggers that are parasites of terrestrial vertebrates, including humans, and some species are of medical importance as vectors of chigger-borne rickettsiosis (scrub typhus), caused by the rickettsia Orientia tsutsugamushi. This paper presents an annotated checklist of 3,013 generally accepted chigger species, together with their distribution by zoogeographic region, and a non-comprehensive list of synonyms. A total of 58 new combinations are proposed by transferring species to different genera, treating some subgenera as genera, or updating current generic names. The checklist updates earlier catalogs of the world chigger fauna, but because the literature on chigger taxonomy is voluminous and scattered, similar, regional monographs were utilized in constructing the classificatory foundation. This list may not contain every species, synonym or taxonomic rearrangement that has been published, but it reflects the most recent arrangement of chigger taxa. Distribution records are primarily drawn from collection sites listed in the original species descriptions, together with subsequently published geographical records.

Carbon loss in the form of CO

Current state-of-the-art decadal ensemble prediction systems are run with an ensemble size of 10 to 40 members, their retrospective forecasts of the past are used to assess the system's prediction skill. Here, we present an attempt for a large ensemble decadal prediction system for the time period 1960-today, with an ensemble size of 80 members, based on the low resolution version of the Max Planck Institute Earth system model (MPI-ESM-LR). The ensemble is forced with CMIP6 conditions and initialized every year in November through a weakly coupled assimilation using atmospheric reanalyses via nudging and observed oceanic temperature and salinity profiles via a 16-member ensemble Kalman filter. To generate ensemble members beyond 16, we use additional physical perturbations at stratospheric height. The analysis of our large ensemble prediction system presented here aims for answering two questions: (1) How does the ensemble mean deterministic prediction skill for global and North Atlantic key climate indices change with ensemble size? (2) How well may the 80-member ensemble serve as a basis for a robust statistical analysis of probabilities of extremes in the North Atlantic sector? Preliminary results for global and regional air surface temperature show that in terms of ensemble mean ACC and full ensemble CPRSS with reference data, the 80-member ensemble leads to similar prediction skill as the 16-member ensemble. This indicates that the additional ensemble members may lead to a better sampling of the distribution of model trajectories, paving the way for a more robust statistical probabilistic analysis.

When unprotected by sea-ice and exposed to the warm air and ocean waves, the Arctic coast erodes and releases organic carbon from permafrost to the surrounding ocean and atmosphere. This release is estimated to deliver similar amounts of organic carbon to the Arctic Ocean as all Arctic rivers combined, at the present-day climate. Depending on the degradation pathway of the eroded material, the erosion of the Arctic coast could represent a positive feedback loop in the climate system, to an extent still unknown. In addition, the organic carbon flux from Arctic coastal erosion is expected to increase in the future, mainly due to surface warming and sea-ice loss. In this work, we aim at addressing the following questions: How is Arctic coastal erosion projected to change in the future? How sensitive is Arctic coastal erosion to climate change?To address these questions, we use a 10-member ensemble of climate change simulations performed with the Max Planck Institute Earth System Model (MPI-ESM) for the Coupled Model Intercomparison Project phase 6 (CMIP6) to make projections of coastal erosion at a pan-Arctic scale. We use a semi-empirical approach to model Arctic coastal erosion, assuming a linear contribution of its thermal and mechanical drivers. The pan-Arctic carbon release due to coastal erosion is projected to increase from 6.9 ± 5.4 TgC/year (mean estimate ± two standard deviations from the distribution of uncertainties) during the historical period (mean over 1850 -1950) to between 13.1 ± 6.7 TgC/year and 17.2 ± 8.2 TgC/year in the period 2081-2100 following an intermediate (SSP2.4-5) and a high-end (SSP5.8-5) climate change scenario, respectively. The sensitivity of the organic carbon release from Arctic coastal erosion to climate warming is estimated to range from 1.52 TgC/year/K to 2.79 TgC/year/K depending on the scenario. Our results present the first projections of Arctic coastal erosion, combining observations and Earth system model (ESM) simulations. This allows us to make first-order estimates of sensitivity and feedback magnitudes between Arctic coastal erosion and climate change, which can lay out pathways for future coupled ESM simulations.

The aim of this secondary analysis was to identify prodynorphin (PDYN) genetic markers moderating the therapeutic response to treatment of cocaine dependence with buprenorphine/naloxone (Suboxone®; BUP).Cocaine-dependent participants (N = 302) were randomly assigned to a platform of injectable, extended-release naltrexone (XR-NTX) and one of three daily medication arms: 4 mg BUP (BUP4), 16 mg BUP (BUP16), or placebo (PLB) for 8 weeks (Parent Trial Registration: Protocol ID: NIDA-CTN-0048, Clinical Trials.gov ID: NCT01402492). DNA was obtained from 277 participants. Treatment response was determined from weeks 3 to 7 over each 1-week period by the number of cocaine-positive urines per total possible urines.In the cross-ancestry group, the PLB group had more cocaine-positive urines than the BUP16 group (P = 0.0021). The interactions of genetic variant × treatment were observed in the rs1022563 A-allele carrier group where the BUP16 group (N = 35) had fewer cocaine-positive urines (P = 0.0006) than did the PLB group (N = 26) and in the rs1997794 A-allele carrier group where the BUP16 group (N = 49) had fewer cocaine-positive urines (P = 0.0003) than did the PLB group (N = 58). No difference was observed in the rs1022563 GG or rs1997794 GG genotype groups between the BUP16 and PLB groups. In the African American-ancestry subgroup, only the rs1022563 A-allele carrier group was associated with treatment response.These results suggest that PDYN variants may identify patients who are best suited to treatment with XR-NTX plus buprenorphine for cocaine use disorder pharmacotherapy.

The global transcriptional response of Escherichia coli to styrene and the potential influence of the exposure source was determined by performing RNA sequencing (RNA-seq) analysis on both styrene-producing and styrene-exposed cells. Relative to an unexposed control, 1,574 and 755 genes showed significant changes in differential expression when styrene was exogenously added versus endogenously produced. Taken together, in both cases, styrene exposure appears to cause both cell envelope and DNA damage, to which cells respond by down-regulating key genes and pathways involved in DNA replication, protein production, and cell wall biogenesis. Among the most significantly up-regulated genes under both exposure modes were those involved in the phage shock protein response (e.g. pspABCDE/G), general stress regulators (e.g. marA, rpoH), and several membrane-altering genes (notably, bhsA, ompR, ldtC), whereas most efflux transporters were, surprisingly, unaffected. Subsequent studies performed with styrene addition, meanwhile, demonstrate how strains lacking ompR (involved in controlling outer membrane (OM) composition/osmoregulation) or any of tolQ, tolA, or tolR (all involved in OM constriction) each displayed over 40 per cent reduced growth relative to wild-type E. coli; differences pointing to the likely importance of these OM-associated processes in native styrene tolerance. Conversely, despite reducing the basal fitness of unexposed cells, overexpression of plsX (involved in phospholipid biosynthesis) led to 70 per cent greater growth when styrene exposed; further suggesting the importance of OM properties in controlling tolerance. Overall, the collective behaviors suggest that, regardless of its source, prolonged exposure to inhibitory styrene levels causes cells to shift from a 'growth mode' to 'survival mode', redistributing cellular resources to fuel native tolerance mechanisms.

Background The use of smartphones and multimedia messaging service (MMS) continues to increase in day to day orthopaedic clinical practice. However, there is limited evidence to support the safe utilisation of MMS. Objectives The aim of this study was to correlate the performance of MMS imaging to picture archiving and communication systems (PACS) imaging within the setting of diagnosis and management of ankle fractures. Methods The ankle fracture radiograph series of 82 consecutive patients were evaluated by five orthopaedic consultant specialists. A questionnaire regarding diagnosis and preferred management was completed separately for each patient using smartphone and PACS images. Statistical analysis was performed using Intraclass Correlation Coefficient (ICC). Results Ankle fracture diagnosis showed strong to excellent correlation both inter- and intraobserver MMS vs PACS when using the Weber (0.815, 0.988), Anatomical (0.858, 0.988), and AO classification systems (0.855, 0.985). MMS was less reliable than PACS in determining many management options. Conclusion The reliability of ankle fracture classification using MMS image viewing was not significantly different to interpretation on PACS workstations. Smartphone use in ankle fracture classification is supported by this study. Smartphone use was less accurate than PACS in devising management plans and future use should be limited to making only initial plans that must be corroberated with PACS and intraoperative findings prior to definitive fixation.

OBJECTIVES/HYPOTHESIS Internal auditory diverticula in adults have been found to exist independent of otosclerosis, and in the presence of otosclerosis. We sought to determine the prevalence of internal auditory canal (IAC) diverticula in a pediatric cohort, to assess whether IAC diverticula are a risk factor for hearing loss, and the co-occurrence of otic capsule hypoattenuation. STUDY DESIGN Retrospective review. METHODS A single-site retrospective review of high-resolution temporal bones computed tomography (CT) scans including the presence and size of diverticula and hypoattenuation of the otic capsule. Demographic, imaging, and audiometric data were collected and descriptively analyzed. Bivariate analysis of collected variables was conducted. Comparisons between sides in unilateral cases were also performed. RESULTS 16/600 (2.7%; 95% CI [2.0%, 3.4%]) were found to have IAC diverticula. Six were bilateral. Thirty-one patients (5.2%) were found to have hypoattenuation of the otic capsule. There were no coincident cases of IAC diverticulum and hypoattenuation of the otic capsule. There was no association between the presence of IAC diverticula and age (P = .13). In six patients with unilateral diverticula, pure tone average (P = .42), and word recognition (P = .27) scores were not significantly different when compared to the normal, contralateral side. CONCLUSIONS The prevalence of IAC diverticula in children is lower than the prevalence in adults. IAC diverticula in children likely represent congenital variants of temporal bone anatomy. Similar to adult populations, there is evidence that IAC diverticula in children are likely not an independent risk factor for hearing loss. LEVEL OF EVIDENCE 4 Laryngoscope, 131:E1683-E1687, 2021.

With the goal of expanding the diversity of tools available for controlling gene expression in cyanobacteria, the T7-RNA polymerase gene expression system from E. coli BL21(DE3) was adapted and systematically engineered for robust function Synechococcus sp. PCC 7002, a fast-growing saltwater strain. Expression of T7-RNA polymerase was controlled via LacI regulation, while functionality was optimized by both further tuning its expression level along with optimizing the translation initiation region of the expressed gene, in this case an enhanced YFP reporter. Under high CO2 conditions, the resulting system displayed a 60-fold dynamic range in expression levels. Furthermore, when maximally induced, T7-RNA polymerase-dependent protein production constituted up to two-thirds of total cellular protein content in Synechococcus sp. PCC 7002. Ultimately, however, this came at the cost of 40% reductions in both biomass and pigmentation levels. Taken together, the developed T7-RNA polymerase gene expression system is effective for controlling and achieving high-level expression of heterologous genes in Synechococcus sp. PCC 7002, making it a valuable tool for cyanobacterial research. KEY POINTS: • Promoter driving T7-RNA polymerase was optimized. • Up to 60-fold dynamic range in expression, depending on CO2 conditions. • Two-thirds of total protein is T7-RNA polymerase dependent.

The PII-like protein SbtB has been identified as a regulator of SbtA, which is one of the key bicarbonate transporters in cyanobacteria. While SbtB from Synechocystis sp. PCC 6803 has previously been shown to be a trimer, a new crystal form is reported here which crystallizes in what is thought to be a non-native tetramer in the crystal, with the C-terminus in an extended conformation. The crystal structure shows the formation of an intermolecular disulfide bond at Cys94 between SbtB monomers, which may stabilize this conformation in the crystal. This motivates the need for future studies to investigate the potential role that the oxidation and reduction of these cysteines may play in the activation and/or function of SbtB.