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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.

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.

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.

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.

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.

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.

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.

Although biological upgrading of lignocellulosic sugars represents a promising and sustainable route to bioplastics, diverse and variable feedstock compositions (e.g., glucose from the cellulose fraction and xylose from the hemicellulose fraction) present several complex challenges. Specifically, sugar mixtures are often incompletely metabolized due to carbon catabolite repression while composition variability further complicates the optimization of co-utilization rates. Benefiting from several unique features including division of labor, increased metabolic diversity, and modularity, synthetic microbial communities represent a promising platform with the potential to address persistent bioconversion challenges. In this work, two unique and catabolically orthogonal Escherichia coli co-cultures systems were developed and used to enhance the production of D-lactate and succinate (two bioplastic monomers) from glucose–xylose mixtures (100 g L–1 total sugars, 2:1 by mass). In both cases, glucose specialist strains were engineered by deleting xylR (encoding the xylose-specific transcriptional activator, XylR) to disable xylose catabolism, whereas xylose specialist strains were engineered by deleting several key components involved with glucose transport and phosphorylation systems (i.e., ptsI, ptsG, galP, glk) while also increasing xylose utilization by introducing specific xylR mutations. Optimization of initial population ratios between complementary sugar specialists proved a key design variable for each pair of strains. In both cases, ∼91% utilization of total sugars was achieved in mineral salt media by simple batch fermentation. High product titer (88 g L–1 D-lactate, 84 g L–1 succinate) and maximum productivity (2.5 g L–1 h–1 D-lactate, 1.3 g L–1 h–1 succinate) and product yield (0.97 g g-total sugar–1 for D-lactate, 0.95 g g-total sugar–1 for succinate) were also achieved.

Arctic coastal erosion experiences pronounced effects from ongoing climate change. The Laptev Sea figures among the Arctic regions with the most severe erosion rates. Here, we use unprecedentedly long records of almost 30 years of annual in-situ coastal erosion rate measurements from Bykovsky Peninsula and Muostakh Island to separate the main modes of variability, which we attribute to large-scale drivers. The first (lower-frequency) and second (higher-frequency) modes are associated with winter sea-ice cover in the Laptev Sea and with the Arctic Oscillation, respectively, which together account for (Formula presented.) of the total observed variance. Arctic coastal erosion has so far been neglected in Earth system models (ESMs). The proposed mechanisms set favorable conditions for coastal erosion at large scales (synoptic to planetary scales), compatible with those represented in modern ESMs. ©2020. American Geophysical Union. All Rights Reserved.

Irrigated corn (L.) in the Great Plains will be increasingly grown under limited irrigation management and greater water stress. Hybrids with drought genetics may decrease the impacts of water stress on yield. The objective of this experiment was to evaluate the effect of drought genetics on corn water stress, water use, grain yield, yield components, and water use efficiency under varying water availability. A 3-yr (2009–2011) study was conducted at Akron, CO in which corn water use, yield, and yield components were measured for isolines of three corn hybrids with and without drought genetics grown under three water treatments (fully irrigated, no irrigation until just prior to tasseling followed by full irrigation, and limited irrigation throughout the growing season). There was no yield loss from the drought genetics under non-water-stressed conditions. There was no effect of drought genetics on water use, yield, or water use efficiency. However, there was a 7% increase in kernels per plant in 2011 and a 5% increase in 1000-kernel weight in 2010 due to the presence of the drought gene. It does not appear that the non-commercial, experimental drought genetics tested in this experiment produced any consistent effect on mitigating water stress in corn or on corn water use and yield under varying levels and timings of water stress. Further research is needed in which consistent levels of water stress are imposed during reproductive and grain-filling growth stages to verify the effects of drought genetics on corn yield.

The winter wheat (Triticum aestivum L.)–fallow (WF) dryland production system employed in the Central Great Plains has evolved in the past 40 yr to include a diversity of other crops, with a reduction in fallow frequency. Wheat remains the base crop for essentially all cropping systems. Decisions to change a farming system benefit from information about average wheat yields, yield stability, and probabilities of obtaining a specified minimum wheat yield. The objective of this experiment was to quantify wheat yields, yield stability, and the probability of obtaining a specified minimum yield in eight dryland rotational systems varying in cropping intensity. The study was conducted over a 24-yr period at Akron, CO. Yield stability was characterized with six stability measures. The probability of obtaining a yield less than 1500 kg ha⁻¹ was also calculated for each rotation. Wheat yields were greatest in rotations where wheat followed a fallow period and least where wheat followed millet production. Rotations ranked from most stable to least stable wheat production (averaged over the six stability measures) were WF(NT), WF(CT), WMF, WCMP, WCM, WCMF, WM, and WCF. The probability of producing

Reported cases of Lyme disease in Nebraska have been assumed to be imported from other endemic areas. Previous surveillance efforts provided no evidence of established populations as only individual specimens of Ixodes scapularis (Say) had been collected. In the winter of 2018, adult I. scapularis were found on a dog at Two Rivers State Recreation Area, Douglas County, prompting tick collection at the site and nearby natural areas. In May 2019, all life stages of host-seeking I. scapularis were collected using dragging and flagging techniques in sites located near the Platte River in Douglas, Sarpy, and Saunders counties. This is the first documentation of established populations of I. scapularis in Nebraska.

Genetic variation in the serotonin transporter (SLC6A4) has been shown to moderate the acute subjective effects of cocaine. Methylation of the SLC6A4 gene is associated with decreased transcription of the serotonin transporter, leading to increased serotonin in the synapse. In this study, methylation of the SLC6A4 gene was investigated in the moderation of the subjective effects of cocaine. Non-treatment-seeking cocaine-dependent individuals (N = 53) were intravenously administered cocaine (40 mg) and saline in a randomized order. The subjective effects of cocaine were self-reported using a visual analog scale starting prior to the administration of cocaine (−15 min) or saline and up to 20 min after infusion. Participants were evaluated for methylation of the SLC6A4 promoter region and 5-HTTLPR genotype. A series of ANCOVAs for SLC6A4 methylation (high/low) were run for each of ten subjective and three cardiovascular effects controlling for age, sex [utilizing the sex-determining region Y protein (SRY)], and population structure (determined from ancestry informative markers and STRUCTURE software). Participants with SLC6A4 hypermethylation reported greater subjective response to cocaine for ‘depressed’ relative to participants with SLC6A4 hypomethylation (experiment-wise p =.002). These findings indicate that SLC6A4 methylation moderates the ‘depressed’ subjective effect of cocaine in non-treatment-seeking cocaine-dependent participants.

Reaction of [CoCp2][TpiPrMoOS(OAr)] [Cp = η5-cyclopentadienyl; TpiPr = hydrotris(3-isopropylpyrazol-1-yl)borate; OAr = phenolate or derivative thereof] with [Cu(NCMe)(Me3tcn)]BF4 (Me3tcn = 1,4,7-trimethyl-1,4,7-triazacyclononane) in MeCN at −30 °C results in the formation of red-brown/black, paramagnetic, μ-sulfido-Mo(V)/Cu(I) complexes, TpiPrMoO(OAr)(μ-S)Cu(Me3tcn). The complexes possess the MoO(μ-S)Cu core found in aerobic carbon monoxide dehydrogenases (CODHs) and exhibit X-band EPR spectra closely related to those of semi-reduced CODH, with giso ∼ 1.937, hyperfine coupling to 95,97Mo (aiso = 39–42 × 10−4 cm−1) and strong superhyperfine coupling to 63,65Cu (aiso = 34–63 × 10−4 cm−1). Anisotropic spectra exhibit monoclinic symmetry with g1 ∼ 1.996, g2 ∼ 1.944 and g3 ∼ 1.882, and nearly isotropic ACu values (75–90 × 10−4 cm−1). The X-ray structures of four derivatives (Ar = Ph, C6H4tBu-2, C6H4sBu-2, C6H4Ph-4) are reported and discussed along with that of the Ar = C6H3tBu2-3,5 derivative (communicated in C. Gourlay, D. J. Nielsen, J. M. White, S. Z. Knottenbelt, M. L. Kirk and C. G. Young, J. Am. Chem. Soc., 2006, 128, 2164). The complexes exhibit distorted octahedral oxo-Mo(V) and distorted tetrahedral Cu(I) centres bridged by a single bent μ-sulfido ligand, with Mo–S and Cu–S distances and Mo–S–Cu angles in the ranges 2.262–2.300 A, 2.111–2.134 A and 115.87–134.27°, respectively. The 2 t-butyl derivative adopts a unique phenolate conformation with OMo–O–Cα and OMo–S–Cu torsion angles of 92.7 and 21.1°, respectively, very different from those of the other structurally characterized derivatives (31–47 and 33–45°, respectively) and exhibits a relatively short Mo⋯Cu distance [3.752(2) A vs. 3.806(7)–4.040(2) A]. As well, the aCu value of this complex (34.3 × 10−4 cm−1) is much lower than the values observed for other members of the series (55–63 × 10−4 cm−1), supporting the hypothesis that the electronic structure of the MoO(μ-S)Cu core unit and the degree of intermetallic communication are strongly dependent on the geometry of the MoO(OR)(μ-S)Cu unit. The complexes participate in an electrochemically reversible Mo(VI)/Mo(V) redox couple and react with cyanide undergoing decupration and desulfurization reactions of the type observed for CODH.

Proso millet ( Panicum miliaceum L.) is a short-season, drought-tolerant C4 species capable of making use of limited available water supplies and is suitable for dryland crop rotations in the Central Great Plains. Previously published water use/yield production functions for proso millet have slopes lower than reported for other C4 species in this region. The objectives of this experiment were to determine the water-limited yield relationship for proso millet and to identify environmental factors that cause yields to be lower than predicted by the water-limited yield relationship. Water use and yield data were obtained from a long-term crop rotation experiment conducted under dryland conditions in northeast Colorado from 1995 to 2016. Stepwise linear regression analysis was used to determine important environmental factors influencing yield. The water-limited yield relationship had a slope consistent with other C4 species in this region (32.57 kg ha −1 per mm of water use). A relationship based on growing season water use, plant available soil water at planting, precipitation received from 12 to 18 August, number days in July and August with maximum temperature greater than 36°C, daily average wind run and maximum wind gust during the week before swathing explained 88% of yield variability. The results of this analysis suggest that closing the yield gap for proso millet production could likely result from efforts to breed for enhanced shattering resistance and heat tolerance and from production methods that improve precipitation storage efficiency during the non-crop period prior to millet planting and increase available soil water at millet planting.

Background and objectives Opioid use disorder (OUD) is a chronic disorder with relapse based on both desire for reinforcement (craving) and avoidance of withdrawal. The aversive aspect of dependence and relapse has been associated with a small brain structure called the habenula, which expresses large numbers of both opioid and nicotinic receptors. Additionally, opioid withdrawal symptoms can be induced in opioid-treated rodents by blocking not only opioid, but also nicotinic receptors. This receptor co-localization and cross-induction of withdrawal therefore might lead to genetic variation in the nicotinic receptor influencing development of human opioid dependence through its impact on the aversive components of opioid dependence. Methods We studied habenular resting state functional connectivity with related brain structures, specifically the striatum. We compared abstinent psychiatric patients who use opioids (N = 51) to psychiatric patients who do not (N = 254) to identify an endophenotype of opioid use that focused on withdrawal avoidance and aversion rather than the more commonly examined craving aspects of relapse. Results We found that habenula-striatal connectivity was stronger in opioid-using patients. Increased habenula-striatum connectivity was observed in opioid-using patients with the low risk rs16969968 GG genotype, but not in patients carrying the high risk AG or AA genotypes. Conclusions We propose that increased habenula-striatum functional connectivity may be modulated by the nicotinic receptor variant rs16969968 and may lead to increased opioid use. Scientific significance Our data uncovered a promising brain target for development of novel anti-addiction therapies and may help the development of personalized therapies against opioid abuse. (Am J Addict 2017;26:751-759).

Microbial systems have been widely studied and exploited through genetic engineering to address industrial needs and societal challenges. However, owing to their complexity, singular approaches often do not yield desired or optimal results, pushing researchers to explore combinatorial strategies. With advances in synthetic biology, various methods can readily be employed to generate large and comprehensive libraries. To serve as tractable tools, however, this capability necessitates the development of high-throughput screening (HTS) techniques to identify the best performing strain and/or those carrying the desired trait. Owing to their miniaturization, time efficiency, potential for automation, and so on, HTS enables comprehensive exploration of diverse experimental landscapes. Herein, we review the recent and novel HTS approaches and applications in the realm of microbial biotechnology.

The South Atlantic Convergence Zone (SACZ) plays a key role in the South American monsoon system (SAMS) precipitation regime, and accounts for around 25 of the rainfall volume over Southeast Brazil between October and April, on average, with peaks of 56 in March and 41 in January. Due to its often varying position and multi-variable structure, diagnosing or quantifying SACZ episodes may lie on subjective criteria. The present study provides a climatological characterization of different SACZ types, based on their position, and differences in dynamics analysed from 19 SAMS wet periods. A cycle in SACZ configurations is identified during the rainy season: northernmost (southernmost) episodes are more likely to occur during the onset/demise (peak) months of the SAMS. Objective SACZ indices are developed taking the principal components of the dynamics of the SACZ as explanatory variables, not including precipitation. The most sensitive threshold proposed for the identification of mean SACZ type yields simultaneous true positive rate of 86, false alarm rate of 28, true negative rate of 72 and false negative rate of 15, for example. The indices have potential to bring gains in predictability to SACZ-related precipitation in Southeast Brazil, apart from allowing the objective diagnosis of SACZ episodes. The indices are also correlated with the South American precipitation dipole and may be used to identify anomalous SAMS precipitation at longer time scales as well. © 2018, The Author(s).

Eleven triatomine species, the vector for Chagas disease, are endemic in the southern U.S. While traditionally thought to only occur in rural habitats and sylvatic transmission cycles, recent studies provide compounding evidence that triatomines could exist in urban habitats and domestic transmission cycles in Texas. We conducted a study of active and passive surveillance techniques over 3 years (2016-2018) in the City of Houston, Harris County, Texas to determine the presence of triatomines in this metroplex. Active surveillance methods uncovered Triatoma sanguisuga nymphs from two locations in downtown Houston city parks. We also documented the first Trypanosoma cruzi positive kissing bug collected in an urban environment of Harris County, Texas. Our findings provide evidence that triatomines can be found in heavily populated U.S. urban environments, and warrant public health support for expanded triatomine and Chagas disease surveillance in city settings.

Efficient microbial conversion of lignocellulose into valuable products is often hindered by the presence of furfural, a dehydration product of pentoses in hemicellulose sugar syrups derived from woody biomass. For a cost-effective lignocellulose microbial conversion, robust biocatalysts are needed that can tolerate toxic inhibitors while maintaining optimal metabolic activities. A comprehensive plasmid-based library encoding native multidrug resistance (MDR) efflux pumps, porins, and select exporters from Escherichia coli was screened for furfural tolerance in an ethanologenic E. coli strain. Small multidrug resistance (SMR) pumps, such as SugE and MdtJI, as well as a lactate/glycolate:H(+) symporter, LldP, conferred furfural tolerance in liquid culture tests. Expression of the SMR pump potentially increased furfural efflux and cellular viability upon furfural assault, suggesting novel activities for SMR pumps as furfural efflux proteins. Furthermore, induced expression of mdtJI enhanced ethanol fermentative production of LY180 in the presence of furfural or 5-hydroxymethylfurfural, further demonstrating the applications of SMR pumps. This work describes an effective approach to identify useful efflux systems with desired activities for nonnative toxic chemicals and provides a platform to further enhance furfural efflux by protein engineering and mutagenesis. IMPORTANCE Lignocellulosic biomass, especially agricultural residues, represents an important potential feedstock for microbial production of renewable fuels and chemicals. During the deconstruction of hemicellulose by thermochemical processes, side products that inhibit cell growth and production, such as furan aldehydes, are generated, limiting cost-effective lignocellulose conversion. Here, we developed a new approach to increase cellular tolerance by expressing multidrug resistance (MDR) pumps with putative efflux activities for furan aldehydes. The developed plasmid library and screening methods may facilitate new discoveries of MDR pumps for diverse toxic chemicals important for microbial conversion.

Background and Objectives There is significant variability in severity of neonatal abstinence syndrome (NAS) due to in utero opioid exposure. Our previous study identified single nucleotide polymorphisms (SNPs) in the prepronociceptin (PNOC) and catechol-O-methyltransferase (COMT) genes that were associated with differences in NAS outcomes. This study looks at the same SNPs in PNOC and COMT in an independent cohort in an attempt to replicate previous findings. Methods For the replication cohort, full-term opioid-exposed newborns and their mothers (n = 113 pairs) were studied. A DNA sample was obtained and genotyped for five SNPs in the PNOC and COMT genes. The association of each SNP with NAS outcomes (length of hospitalization, need for pharmacologic treatment, and total opioid days) was evaluated, with an experiment-wise significance level set at α

Epigenetic processes regulate gene expression independent of the DNA sequence and are increasingly being investigated as contributors to the development of behavioral disorders. Environmental insults, such as stress, diet, or toxin exposure, can affect epigenetic mechanisms, including chromatin remodeling, DNA methylation, and non-coding RNAs that, in turn, alter the organism's phenotype. In this review, we examine the literature, derived at both the preclinical (animal) and clinical (human) levels, on epigenetic alterations associated with anxiety disorders. Using animal models of anxiety, researchers have identified epigenetic changes in several limbic and cortical brain regions known to be involved in stress and emotion responses. Environmental manipulations have been imposed prior to conception, during prenatal or early postnatal periods, and at juvenile and adult ages. Time of perturbation differentially affects the epigenome and many changes are brain region-specific. Although some sex-dependent effects are reported in animal studies, more research employing both sexes is needed particularly given that females exhibit a disproportionate number of anxiety disorders. The human literature is in its infancy but does reveal some epigenetic associations with anxiety behaviors and disorders. In particular, effects in monoaminergic systems are seen in line with evidence from etiological and treatment research. Further, there is evidence that epigenetic changes may be inherited to affect subsequent generations. We speculate on how epigenetic processes may interact with genetic contributions to inform prevention and treatment strategies for those who are at risk for or have anxiety disorders.

Targeted delivery of anticancer therapeutics can potentially overcome the limitations associated with current chemotherapeutic regimens. Folate receptors are overexpressed in several cancers, including ovarian, triple‐negative breast and bladder cancers, making them attractive for targeted delivery of nucleic acid therapeutics to these tumors. This work describes the synthesis, characterization and evaluation of folic acid‐conjugated, aminoglycoside‐derived polymers for targeted delivery of transgenes to breast and bladder cancer cell lines. Transgene expression was significantly higher with FA‐conjugated aminoglycoside‐derived polymers than with Lipofectamine, and these polymers demonstrated minimal cytotoxicty. Competitive inhibition using free folic acid significantly reduced transgene expression efficacy of folate‐targeted polymers, suggesting a role for folate receptor‐mediated uptake. High efficacy FA‐targeted polymers were employed to deliver a plasmid expressing the TRAIL protein, which induced death in cancer cells. These results indicate that FA‐conjugated aminoglycoside‐derived polymers are promising for targeted delivery of nucleic acids to cancer cells that overexpress folate receptors.

Constructing microbial biocatalysts that produce biorenewables at economically viable yields and titers is often hampered by product toxicity. For production of short chain fatty acids, membrane damage is considered the primary mechanism of toxicity, particularly in regards to membrane integrity. Previous engineering efforts in Escherichia coli to increase membrane integrity, with the goal of increasing fatty acid tolerance and production, have had mixed results. Herein, a novel approach was used to reconstruct the E. coli membrane by enabling production of a novel membrane component. Specifically, trans unsaturated fatty acids (TUFA) were produced and incorporated into the membrane of E. coli MG1655 by expression of cis-trans isomerase (Cti) from Pseudomonas aeruginosa. While the engineered strain was found to have no increase in membrane integrity, a significant decrease in membrane fluidity was observed, meaning that membrane polarization and rigidity were increased by TUFA incorporation. As a result, tolerance to exogenously added octanoic acid and production of octanoic acid were both increased relative to the wild-type strain. This membrane engineering strategy to improve octanoic acid tolerance was found to require fine-tuning of TUFA abundance. Besides improving tolerance and production of carboxylic acids, TUFA production also enabled increased tolerance in E. coli to other bio-products, e.g. alcohols, organic acids, aromatic compounds, a variety of adverse industrial conditions, e.g. low pH, high temperature, and also elevated styrene production, another versatile bio-chemical product. TUFA permitted enhanced growth due to alleviation of bio-product toxicity, demonstrating the general effectiveness of this membrane engineering strategy towards improving strain robustness.

Fermentative production of styrene from glucose has been previously demonstrated in Escherichia coli. Here, we demonstrate the production of styrene from the sugars derived from lignocellulosic biomass depolymerized by fast pyrolysis. A previously engineered styrene-producing strain was further engineered for utilization of the anhydrosugar levoglucosan via expression of levoglucosan kinase. The resulting strain produced 240 ± 3 mg L−1 styrene from pure levoglucosan, similar to the 251 ± 3 mg L−1 produced from glucose. When provided at a concentration of 5 g L−1, pyrolytic sugars supported styrene production at titers similar to those from pure sugars, demonstrating the feasibility of producing this important industrial chemical from biomass-derived sugars. However, the toxicity of contaminant compounds in the biomass-derived sugars and styrene itself limit further gains in production. Styrene toxicity is generally believed to be due to membrane damage. Contrary to this prevailing wisdom, our quantitative assessment during challenge with up to 200 mg L−1 of exogenously provided styrene showed little change in membrane integrity; membrane disruption was observed only during styrene production. Membrane fluidity was also quantified during styrene production, but no changes were observed relative to the non-producing control strain. This observation that styrene production is much more damaging to the membrane integrity than challenge with exogenously supplied styrene provides insight into the mechanism of styrene toxicity and emphasizes the importance of verifying proposed toxicity mechanisms during production instead of relying upon results obtained during exogenous challenge.

Background and objectives Disulfiram has been beneficial in treating cocaine addiction in several studies. Patients with two SLC6A3 (DAT1) rs28363170 10-repeat alleles who have with genetically high dopamine transporter (DAT) levels may benefit from increased dopamine levels resulting from disulfiram treatment. Methods After stabilization for 2 weeks on methadone, 70 cocaine and opioid codependent patients were randomized into disulfiram and placebo groups for 12 weeks of treatment. We genotyped the SLC6A3 (DAT1) 40 bp 3'-untranslated region variable number tandem repeat variant and evaluated its role in moderating disulfiram efficacy for cocaine dependence. Results Among the 10,10-repeat genotype group, cocaine-positive urines dropped from 78% to 48% and from 80% to 75% among the 9-repeat carrier group in the disulfiram group (P = 0.0001, with an effect size of 0.09). No difference was observed in cocaine-positive urines in the placebo group between the 10,10-repeat genotype and the 9-allele carrier patients. Conclusions and scientific significance We found that patients with genetically higher DAT levels had better treatment outcomes with disulfiram pharmacotherapy of cocaine dependence than those with lower DAT levels. (Am J Addict 2019;28:311-317).

BACKGROUND: The α(1) adrenergic antagonist, doxazosin, has improved cocaine use disorder (CUD) presumably by blocking norepinephrine (NE) stimulation and reward from cocaine-induced NE increases. If the NE levels for release were lower, then doxazosin might more readily block this NE stimulation and be more effective. The NE available for release can be lower through a genetic polymorphism in dopamine β-hydroxylase (DBH) (C-1021T, rs1611115), which reduces DβH’s conversion of dopamine to NE. We hypothesize that doxazosin would be more effective in CUD patients who have these genetically lower DβH levels. METHODS: This 12-week, double-blind, randomized, placebo-controlled trial included 76 CUD patients: 49 with higher DβH levels from the DBH CC genotype, and 27 with lower DβH levels from T-allele carriers (CT or TT). Patients were randomized to doxazosin (8 mg/day, N=47) or placebo (N=29), and followed with thrice weekly urine toxicology and once weekly cognitive behavioral psychotherapy. RESULTS: Cocaine use was reduced at a higher rate among patients in the doxazosin than in the placebo arm. We found significantly lower cocaine use rates among patients carrying the T-allele (CT/TT) than the CC genotype. The percentage of cocaine positive urines was reduced by 41% from baseline in the CT/TT group with low DβH and NE levels, as compared to no net reduction in the CC genotype group with normal DβH and NE levels. CONCLUSIONS: The DBH polymorphism appears play an important role in CUD patients’ response to doxazosin treatment, supporting a pharmacogenetic association and potential application for personalized medicine.