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The dopamine transporter (DAT) is targeted in substance use disorders (SUDs), and "non-classical”" DAT inhibitors with low abuse potential are therapeutic candidates. Lobinaline, from Lobelia cardinalis, is an atypical DAT inhibitor lead. Chemical synthesis of lobinaline is challenging; thus, "target-directed evolution" was used for lead optimization. A target protein is expressed in plant cells, and a mutant cell population is selected under conditions where target protein functional inhibition confers a survival advantage. Surviving mutants are "mined" for the targeted activity. Applied to a mutant L. cardinalis cell population expressing the human DAT, we identified 20 mutants overproducing DAT inhibitors. Microanalysis prioritized novel lobinaline derivatives, and we first investigated the more water-soluble lobinaline N-oxide. It inhibited rat synaptosomal [3H]DA uptake with an IC50 similar to lobinaline. Against repeated DA microinjections into the rat striatum, lobinaline produced transient DA clearance reductions. In contrast, lobinaline N-oxide prolongingly increased DA peak amplitudes, particularly in the ventral striatum. Lobinaline N-oxide also produced complex changes in post-peak DA clearance inconsistent with simple DAT inhibition. This unusual DAT interaction may prove therapeutically useful for treating SUDs. This study demonstrates the value of target-directed evolution of plant cells for optimizing lead compounds difficult to synthesize chemically.

BackgroundIn gender-affirming medical care (GAMC), ethical challenges in decision-making are ubiquitous. These challenges are becoming more pressing due to exponentially increasing referrals, politico-legal contestation, and divergent normative views regarding decisional roles and models. Little is known, however, about what ethical challenges related to decision-making healthcare professionals (HCPs) themselves face in their daily work in GAMC and how these relate to, for example, the subjective nature of Gender Incongruence (GI), the multidisciplinary character of GAMC and the role HCPs play in assessing GI and eligibility for interventions. Given the relevance and urgency of these questions, we conducted a qualitative study among HCPs providing GAMC to transgender adults in the Netherlands.MethodsIn this qualitative research, we conducted 11 semi-structured interviews between May 2020 and February 2021 with HCPs (six mental health professionals, two HCPs in endocrinology, two in plastic surgery, and one in nursing) working in two distinct GAMC settings. We purposively sampled for professional background and years of experience in GAMC. We analyzed our interview data using thematic analysis. As some respondents were more inclined to speak about what should or ought to be done to arrive at good or right decision-making, we identified both ethical challenges and norms. Furthermore, in our analysis, we differentiated between respondents’ explicit and implicit ethical challenges and norms and ascertained the specific context in which these challenges emerged.ResultsRespondents’ ethical challenges and norms centered on (1) dividing and defining decisional roles and bounds, (2) negotiating decision-making in a (multidisciplinary) team, and (3) navigating various decision-making temporalities. These themes arose in the context of uncertainties regarding (1) GAMC’s guidelines, evidence, and outcomes, as well as (2) the boundaries and assessment of GI.ConclusionsThis interview study provides detailed empirical insight into both the explicit and implicit ethical challenges that HCPs experience and their ethical norms regarding decision-making. It also describes how uncertainties and (implicit) normativities concerning GAMC and GI pre-structure the moral environment in which these challenges and norms manifest. We provide normative reflections and recommendations on handling these ethical challenges in a way that is sensitive to the context in which they arise.

Over the past decades, great strides have been made to professionalize and increase access to transgender medicine. As the (biomedical) evidence base grows and conceptualizations regarding gender dysphoria/gender incongruence evolve, so too do ideas regarding what constitutes good treatment and decision-making in transgender healthcare. Against this background, differing care models arose, including the ‘Standards of Care’ and the so-called ‘Informed Consent Model’. In these care models, ethical notions and principles such as ‘decision-making’ and ‘autonomy’ are often referred to, but left unsubstantiated. This not only transpires into the consultation room where stakeholders are confronted with many different ethical challenges in decision-making, but also hampers a more explicit discussion of what good decision-making in transgender medicine should be comprised of. The aim of this paper is to make explicit the conceptual and normative assumptions regarding decision-making and client autonomy underpinning the ‘Standards of Care’ and ‘Informed Consent Model’ currently used in transgender care. Furthermore, we illustrate how this elucidation aids in better understanding stakeholders’ ethical challenges related to decision-making. Our ethical analysis lays bare how distinct normative ambiguities in both care models influence decision-making in practice and how foregrounding one normative model for decision-making is no moral panacea. We suggest that the first steps towards good decision-making in gender-affirming medical care are the acknowledgement of its inherent normative and moral dimensions and a shared, dialogical approach towards the decision-making process.

Lignin and lignans are natural products found in plant cell walls. Lignin research has historically focused on lignin degradation techniques in the hope of converting lignin into useful aromatic carbon feedstocks. In contrast, investigations of lignans existing as natural product dimers, have been focused on thier interesting biological activities. Many lignan compounds are chemically identical to dimers derived from lignin, and both lignin and lignan dimers can possess multiple chiral centers leading to observations of diastereomer pairs where one diastereomer exhibits the bulk of the activity. For example, the G-(β-O-4′)-G dimer was reported to have a pro-angiogenic activity with one diastereomer of the pair showing enhanced pro-angiogenic activity. Traditional analytical techniques such as nuclear magnetic resonance (NMR) can differentiate the diastereomer pairs of β-O-4 compounds; however, isolation of a pure sample is often required for analysis. This work was aimed at exploring the potential use of tandem mass spectrometry to differentiate diastereomer pairs in the β-O-4 dimer series. Each diastereomer pair in the nine-dimer series was separated by HPLC and interrogated by tandem mass spectrometry. To understand the chromatographic behavior of the diastereomer pair in the β-O-4 dimer series, three commercially available reverse phase HPLC columns were evaluated. A temperature programming experiment using water/acetonitrile isocratic elution showed that the chromatographic retention mechanism of these diastereomers was hydrophobically driven with analytes having more methoxy groups exhibiting larger ΔH0 and higher octanol-water partition coefficient values. Tandem mass spectrometry performed on each of the diastereomers produced fragment ions having different ion abundances. A mechanistic study based on the ion abundance of “sequence-specific ions” and “-48 ions” was used to assign a configuration to each of the pairs of diastereomers in the nine-dimer series.

Lactic acid bacteria (LAB) contamination during fuel ethanol fermentation can lead to significant economic loses. To circumvent this, fuel ethanol plants add antibiotics prophylactically, but their overuse has resulted in the emergence of antibiotic-resistant LAB strains. Lignin is a sustainable biopolymer that can be found as a waste product from lignocellulosic biorefineries. Technical lignins and their smaller phenolic subunits have been shown to exhibit broad-spectrum antimicrobial properties, but there is a lack of demonstrations of lignin derivatives with highly selective properties in the literature. Here, corn stover lignin from a biorefinery was oxidatively depolymerized using an environmentally benign organic oxidant, peracetic acid, into a bio-oil that has selective antimicrobial properties against LAB and not yeasts. The resulting bio-oil demonstrated up to 90% inhibition of commercially sampled LAB (including antibiotic-resistant strains) at 4 mg ml−1 with no inhibition against an industrial yeast strain. These antimicrobial properties of the bio-oil are attributed to larger unidentified lignin oligomers, compared to monolignols, that have a membrane damaging mode of action. Using the bio-oil (4 mg ml−1) during simultaneous saccharification and fermentation (SSF) of raw corn starch showed no inhibition of enzymatic activity, and in LAB contaminated fermentations the bio-oil treatments showed an 8% increase in ethanol yields at higher bacterial contamination ratios (l : 100 yeast to LAB, CFU per ml). This study illustrates the efficacy of using lignin bio-oil as an antibiotic replacement during fuel ethanol fermentation and demonstrates the highly selective antimicrobial properties of lignin oligomers, which creates a viable lignin valorization strategy for biorefineries.

Converting lignin to value-added products at high yields provides an avenue for making ethanol biorefineries more profitable while reducing the carbon footprint of products generally derived from petroleum. In this study, corn stover lignin was depolymerized by catalytic transfer hydrogenolysis (CTH) in supercritical ethanol with a Ru/C catalyst. The lignin-derived bio-oil was then sequentially extracted utilizing hexane, petroleum ether, chloroform, and ethyl acetate as solvents in order of less polar to polar, and the subsequent bio-oils were characterized using GPC, GC/MS, and HSQC NMR. Results show that the monomers in the bio-oil fractions contained primarily alkylated phenols, hydrogenated hydroxycinnamic acid derivatives, syringol and guaiacol-type lignins created from reductive cleavages of ether linkages, which were sequentially extracted into groups depending on the solvent polarity. The antimicrobial properties of the bio-oils were screened against Gram-positive (Bacillus subtilis, Lactobacillus amylovorus, and Staphylococcus epidermidis) and Gram-negative (Escherichia coli) bacteria and yeast (Saccharomyces cerevisiae) by examining microbial growth inhibition. Results show that CTH-derived bio-oils inhibited all tested organisms at concentrations less than 3 mg/mL. Total monomer concentration and the presence of specific monomers (i.e., syringyl propane) showed correlations to antimicrobial activity, likely due to cell death or membrane damage. This study provides insights into using sequential extraction to fractionate lignin-derived compounds and correlations between the properties of the extracted compounds and their antimicrobial activity.

Pyrolysis of hydrolytic lignin (HL) in newly designed, gas phase continuous droplet evaporation (CDE) and continuous atomization (CA) reactors was studied. The product distribution was strongly dep...

Structural elucidation of lignin degradation products is a requirement for successfully developing lignin valorization technology. Most of mass spectrometry-based techniques have utilized negative ion mode mass spectrometry for structural elucidation of lignin-derived compounds. Unfortunately, simple deprotonation can lead to in-source fragmentation and may not be suitable for condensed lignin structures without acidic moieties. Herein, we present a lithium cationization methodology for mass spectrometry sequencing of advanced lignin oligomers having β-β' and β-O-4' bonding motifs. To do so, two advanced lignin oligomers were first synthesized through a step-by-step synthetic route, and then subjected to two different ESI mass spectrometry techniques in positive ion mode using lithium cations for ionization. An orbitrap mass spectrometer was used to obtain exact mass information, and higher-energy collisional dissociation (HCD) was used to sequence the lignin model oligomers. Based on the sequence-specific fragment ions, sequence rules were proposed. Multi-stage (MS

Lignin derived from lignocellulosic biomass is the largest source of renewable bioaromatics present on earth and requires environmentally sustainable separation strategies to selectively obtain high-value degradation products. Applications of supramolecular interactions have the potential to isolate lignin compounds from biomass degradation fractions by the formation of variable inclusion complexes with cyclodextrins (CDs). CDs are commonly used as selective adsorbents for many applications and can capture guest molecules in their internal hydrophobic cavity. The strength of supramolecular interactions between CDs and lignin model compounds that represent potential lignocellulosic biomass degradation products can be characterized by assessing the thermodynamics of binding stability. Consequently, the inclusion interactions of β-CD and lignin model compounds G-(β-O-4')-G, G-(β-O-4')

Three-dimensional matrices of collagen type I (Col I) are widely used in tissue engineering applications for its abundance in many tissues, bioactivity with many cell types, and excellent biocompatibility. Inspired by the structural role of lignin in a plant tissue, we found that sodium lignosulfonate (SLS) and an alkali-extracted lignin from switchgrass (SG) increased the stiffness of Col I gels. SLS and SG enhanced the stiffness of Col I gels from 52 to 670 Pa and 52 to 320 Pa, respectively, and attenuated shear-thinning properties, with the formulation of 1.8 mg/mL Col I and 5.0 mg/mL SLS or SG. In 2D cultures, the cytotoxicity of collagen-SLS to adipose-derived stromal cells was not observed and the cell viability was maintained over 7 days in 3D cultures. Collagen-SLS composites did not elicit immunogenicity when compared to SLS-only groups. Our collagen-SLS composites present a case that exploits lignins as an enhancer of mechanical properties of Col I without adverse cytotoxicity and immunogenicity for

Surface guidance systems enable patient positioning and motion monitoring without using ionising radiation. Surface Guided Radiation Therapy (SGRT) has therefore been widely adopted in radiation therapy in recent years, but guidelines on workflows and specific quality assurance (QA) are lacking. This ESTROACROP guideline aims to give recommendations concerning SGRT roles and responsibilities and highlights common challenges and potential errors. Comprehensive guidelines for procurement, acceptance, commissioning, and QA of SGRT systems installed on computed tomography (CT) simulators, C-arm linacs, closed-bore linacs, and particle therapy treatment systems are presented that will help move to a consensus among SGRT users and facilitate a safe and efficient implementation and clinical application of SGRT. (c) 2022 The Author(s). Published by Elsevier B.V. Radiotherapy and Oncology xxx (2022) xxx-xxx This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

Successful strategies for the attachment of oligopeptides to mesoporous silica with pores large enough to load biomolecules should utilize the high surface area of pores to provide an accessible, protective environment. A two-step oligopeptide functionalization strategy is examined here using diazirine-based heterobifunctional linkers. Mesoporous silica nanoparticles (MSNPs) with average pore diameter of ~8 nm and surface area of ~730 m2/g were synthesized and amine-functionalized. Tetrapeptides Gly-Gly-Gly-Gly (GGGG) and Arg-Ser-Ser-Val (RSSV), and a peptide comprised of four copies of RSSV (4RSSV), were covalently attached via their N-terminus to the amine groups on the particle surface by a heterobifunctional linker, sulfo-succinimidyl 6-(4,4′-azipentanamido)hexanoate (sulfo-NHS-LC-diazirine, or SNLD). SNLD consists of an amine-reactive NHS ester group and UV-activable diazirine group, providing precise control over the sequence of attachment steps. Attachment efficiency of RSSV was measured using fluorescein isothiocyanate (FITC)-tagged RSSV (RSSV-FITC). TGA analysis shows similar efficiency (0.29, 0.31 and 0.26 mol peptide/mol amine, respectively) for 4G, RSSV and 4RSSV, suggesting a generalizable method of peptide conjugation. The technique developed here for the conjugation of peptides to MSNPs provides for their attachment in pores and can be translated to selective peptide-based separation and concentration of therapeutics from aqueous process and waste streams.

Sensitive, high-throughput methods for pharmacokinetic (PK) profiling are essential for potential therapeutics during critical stages of clinical trials. The application of a microfluidic capillary zone electrophoresis mass spectrometry (CZE-MS) method for PK profiling allows for rapid, sensitive and in-depth analysis of multiple samples within a short timeframe. Here, a CZE-MS approach for PK analysis was compared with a traditional UHPLC-MS approach when analyzing serum extracts from rats treated with a potential Alzheimer's disease therapeutic, BNC-1. Resulting PK data generated from both methods displayed statistical similarities. Additionally, the separation efficiency attributed to the use of the CZE-MS method provided substantial metabolic regulation data that was not apparent in the UHPLC-MS method. Additionally, the coupling of the CZE-MS method to the data processing software, MZmine2, was used to monitor changes in metabolism and observe putative BNC-1-derived metabolites. The ability to perform fast analyses without sacrificing sensitivity or metabolic information suggests that this CZE-MS method is ideal for metabolomics-inclusive, high-throughput PK profiling.

A study of the interaction between cell membranes and small molecules derived from lignin, a protective phenolic biopolymer found in vascular plants, is crucial for identifying their potential as pharmacological and toxicological agents. In this work, the interactions of model cell membranes [supported 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) lipid bilayers] are compared for three βO4 dimers of coniferyl alcohol (G lignin monomer): guaiacylglycerol guaiacol ester with a hydroxypropenyl (HOC3H4-) tail (G-βO4′-G), a truncated GG dimer without HOC3H4- (G-βO4′-truncG), and a benzylated GG dimer (benzG-βO4′-G). The uptake of the lignin dimers (per mass of lipid) and the energy dissipation (a measure of bilayer disorder) are higher for benzG-βO4′-G and G-βO4′-truncG than those for G-βO4′-G in the gel-phase DPPC bilayer, as measured using quartz crystal microbalance with dissipation (QCM-D). A similar uptake of G-βO4′-truncG is observed for a fluid-phase bilayer of 1,2-dioleoyl-sn-glycero-3-phosphocholine, suggesting that the effect of the bilayer phase on dimer uptake is minimal. The effects of increasing lignin dimer concentration are examined through an analysis of density profiles, potential of mean force curves, lipid order parameters, and bilayer area compressibilities (disorder) in the lipid bilayers obtained from molecular dynamics simulations. Dimer distributions and potentials of mean force indicate that the penetration into bilayers is higher for benzG-βO4′-G and G-βO4′-truncG than that for G-βO4′-G, consistent with the QCM-D results. Increased lipid tail disorder due to dimer penetration leads to a thinning and softening of the bilayers. Minor differences in the structure of lignin derivatives (such as truncating the hydroxypropenyl tail) have significant impacts on their ability to penetrate lipid bilayers.

High resolution differential scanning calorimetry (DSC) and molecular dynamics (MD) simulations were used to investigate the effect of three lignin dimers on the gel to fluid phase transition in DPPC lipid bilayers. The goal of this research is to begin to understand the partitioning of model lignin dimers into lipid bilayers and its effects on the gel to fluid transition temperature (Tm). The long-term objective is to establish structure-function relationships for well-defined lignin derivatives at biologically relevant surfaces. This work uses a newly synthesized guiacylglycerol guaiacol ester with a hydroxypropenyl (HOC3H4-) group resembling natural lignin (GG dimer), compared with a truncated GG dimer without the HOC3H4- and benzyl-modified GG dimers. The DSC results show that the dimer most like natural lignin (with a hydroxypropenyl tail) has log K = 2.72 ± 0.05, and MD simulations show that it associates with the headgroups of the lipid but does not penetrate strongly into the interior of the bilayer. Therefore, this dimer has little effect on the Tm value. In contrast, the truncated dimer, which has been used as a representative GG dimer in prior studies, partitions into the bilayer, as seen in MD simulations, and shifts Tm because of its increased lipophilicity (DSC log K = 3.45 ± 0.20). Similarly, modification of the natural GG dimer by benzylation of the phenol makes it lipophilic (DSC log K = 3.38 ± 0.28), causing it to partition into the bilayer, as seen in MD simulations and shift Tm. In MD, we capture the transition from gel to fluid phase by defining and analyzing a normalized deuterium order parameter averaged over all carbon atoms located in the middle of the lipid tails. In this way, the phase transition can be clearly observed and, importantly, MD results show the same trend of transition temperature shifts as the DSC results. Furthermore, we compare partition coefficients estimated from free energy profiles calculated in MD to those obtained from experiment and they are in qualitative agreement. The success at predicting the structural effects of lignin dimers on lipid bilayers suggests that MD simulations can be used in the future to screen the interactions of lignin oligomers and their derivatives with lipid bilayers.

Lignin is a crucial component of plant matter; however, it is also largely responsible for the recalcitrance of lignocellulosic biomass when subjected to pretreatment processes. Lignin is generated...

Mass spectrometric analysis of lignin for developing biomaterials requires advances of characterization techniques. Positive ion mass spectrometry of lignin model compounds using lithium has recently been explored as a viable alternative to current negative mode techniques. To date, little is known about the impact of lithium adduct ion formation on relative response factors of lignin and lignin decomposition products. In this contribution, we report estimates of lithium cation basicity for synthetic monolignols H, G and S using Cooks’ kinetic method on a linear quadrupole ion trap mass spectrometer. Optimized geometries and interaction energies have also been calculated by DFT methods to quantify the electrostatic cation coordination. Based on a combination of experimental and computational evidence, lithium appears to preferentially bind to the phenol and methoxy substituents on the aromatic ring of monolignols. The strength of this interaction increases with the number of methoxy substituents (S > G > H). This work serves as a basis of understanding for future work in developing lithium adducted lignin mass spectrometric analytical methods.

RATIONALE Effective analytical techniques are needed to characterize lignin products for the generation of renewable carbon sources. Application of matrix-assisted laser desorption/ionization (MALDI) in lignin analysis is limited because of poor ionization efficiency. In this study, we explored the potential of cationization along with a 2,5-dihydroxyacetophenone (DHAP) matrix to characterize model lignin oligomers. METHODS Synthesized lignin oligomers were analyzed using the developed MALDI method. Two matrix systems, DHAP and α-cyano-4-hydroxycinnamic acid (CHCA), and three cations (lithium, sodium, silver) were evaluated using a Bruker UltraFlextreme time-of-flight mass spectrometer. Instrumental parameters, cation concentration, matrix, sample concentrations, and sample spotting protocols were optimized for improved results. RESULTS The DHAP/Li+ combination was effective for dimer analysis as lithium adducts. Spectra from DHP and ferric chloride oligomers showed improved signal intensities up to decamers (m/z 1823 for the FeCl3 system) and provided insights into differences in the oligomerization mechanism. Spectra from a mixed DHP oligomer system containing H, G, and S units showed contributions from all monolignols within an oligomer level (e.g. tetramer level). CONCLUSIONS The DHAP/Li+ method presented in this work shows promise to be an effective analytical tool for lignin analysis by MALDI and may provide a tool to assess lignin break-down efforts facilitating renewable products from lignin.

The prospect of developing new bio-based products from lignin partially depends on the development of effective analytical techniques to characterize the end products from lignin degradation experiments. To date, the most utilized mass spectrometric technique for characterizing lignin oligomers has been negative ion mass spectrometry. Positive ion mass spectrometry remains a relatively unexplored approach for lignin sequencing. Here we report on the sequencing of lignin oligomers using lithium cationization positive ion electrospray followed by higher-energy collisional dissociation (HCD) tandem mass spectrometry on a high-resolution accurate-mass Orbitrap mass spectrometer. Thirteen synthetic lignin model oligomers containing β-O-4 and 4-O-α linkages were analyzed and sequenced based on their HCD tandem mass spectrometry fragmentation patterns. Proposed sequence-specific fragmentation pathways are provided. The method presented in this study provides a potential application of lithium cationization tandem mass spectrometry for the analysis of lignin degradation products.

Background The majority of hepatitis C virus (HCV) infections are found in low- and middle-income countries, which harbor many region-specific HCV subtypes. Nevertheless, direct-acting antiviral (DAA) trials have almost exclusively been conducted in high-income countries, where mainly epidemically spread HCV subtypes are present. Recently, several studies have demonstrated suboptimal DAA efficacy for certain nonepidemic subtypes, which could hamper global HCV elimination. Therefore, we aimed to evaluate DAA efficacy in patients treated for a nonepidemic HCV genotype infection in the Netherlands. Methods We performed a nationwide retrospective study including patients treated with interferon-free DAAs for an HCV genotype other than 1a/1b/2a/2b/3a/4a/4d. The genotype was determined by NS5B region phylogenetic analysis. The primary end point was SVR-12. If stored samples were available, NS5A and NS5B sequences were obtained for resistance-associated substitutions (RAS) evaluation. Results We included 160 patients, mainly infected with nonepidemic genotype 2 (41%) and 4 (31%) subtypes. Most patients were from Africa (45%) or South America (24%); 51 (32%) were cirrhotic. SVR-12 was achieved in 92% (140/152) of patients with available SVR-12 data. Only 73% (8/11) genotype 3–infected patients achieved SVR-12, the majority being genotype 3b patients with 63% (5/8) SVR. Regardless of SVR, all genotype 3b patients had 30K and 31M RAS. Conclusions The DAA efficacy we observed in most nonepidemic genotypes in the Netherlands seems reassuring. However, the low SVR-12 rate in subtype 3b infections is alarming, especially as it is common in several HCV-endemic countries. Alongside earlier results, our results indicate that a remaining challenge for global HCV elimination is confirming and monitoring DAA efficacy in nonepidemic genotypes.

Background Glaucomatous eyes often show strong intraocular pressure (IOP) fluctuations and individual measurements at different time points are necessary for personalized therapy. To survey IOP variations 48-hours diurnal and nocturnal IOP measurements were performed on two consecutive days. Aims of this study were to investigate the short-term repeatability of 48-hours measurements within one patient’s IOP profile and long-term repeatability between two separate IOP profiles of the same patient. Methods A retrospective cohort study was performed evaluating data of 90 glaucoma patients in a German university medical center between 2006 and 2013. All patients underwent two separate diurnal IOP profiles of 48 h. IOP was measured at 8 am, 2 pm, 6 pm, 9 pm using Goldmann applanation tonometry and at 12 midnight using Perkins tonometry in supine position on two consecutive days. Intraclass correlation coefficients (ICC) were calculated to evaluate agreement for the same time points (each time point agreement) and for consecutive measurements within the IOP profiles (between time point agreement). ICC ≤ 0.4 was defined as poor agreement, 0.4–0.75 as moderate and ≥ 0.75 as excellent. Differences between time points were investigated by Bland Altman plots. Results Each time point measurements of profile 1 showed moderate to excellent agreement (ICCs 0.62–0.93). There was a moderate to excellent agreement for measurements between time points of profile 1 (ICCs day one 0.57–0.86, day two 0.71–0.90). Profile 2 was performed at a median interval of 12.0 months (quartiles 11.0 to 21.0). Each time point agreements within profile 2 showed ICCs from 0.23 to 0.81. It showed moderate to excellent agreement for changes between time points (ICCs 0.53–0.94). Day two demonstrated ICCs from 0.74 to 0.88. Long term IOP repeatability (over both pressure profiles) showed moderate to good agreement (ICCs 0.39–0.67). Conclusions Short and long-term agreement of IOP measurements evaluated by diurnal IOP profiles is moderate to good. Due to mostly moderate agreements, which we believe represent IOP fluctuations, we conclude that it is necessary to perform 48-hours IOP profiles to gain a better overview of the individual IOP fluctuations.

The slow solar wind is typically characterized as having low Alfv��nicity. However, Parker Solar Probe (PSP) observed predominately Alfv��nic slow solar wind during several of its initial encounters. From its first encounter observations, about 55.3\% of the slow solar wind inside 0.25 au is highly Alfv��nic ($|��_C| > 0.7$) at current solar minimum, which is much higher than the fraction of quiet-Sun-associated highly Alfv��nic slow wind observed at solar maximum at 1 au. Intervals of slow solar wind with different Alfv��nicities seem to show similar plasma characteristics and temperature anisotropy distributions. Some low Alfv��nicity slow wind intervals even show high temperature anisotropies, because the slow wind may experience perpendicular heating as fast wind does when close to the Sun. This signature is confirmed by Wind spacecraft measurements as we track PSP observations to 1 au. Further, with nearly 15 years of Wind measurements, we find that the distributions of plasma characteristics, temperature anisotropy and helium abundance ratio ($N_��/N_p$) are similar in slow winds with different Alfv��nicities, but the distributions are different from those in the fast solar wind. Highly Alfv��nic slow solar wind contains both helium-rich ($N_��/N_p\sim0.045$) and helium-poor ($N_��/N_p\sim0.015$) populations, implying it may originate from multiple source regions. These results suggest that highly Alfv��nic slow solar wind shares similar temperature anisotropy and helium abundance properties with regular slow solar winds, and they thus should have multiple origins., submitted to ApJS, welcome comments

The need for renewable bioenergy sources has renewed interest in lignin chemistry; however, structural elucidation and characterization of lignin degradation products remain a challenge because of lack of effective analytical methods. The analysis of lignin oligomers has been accomplished by simple deprotonation of weakly acidic phenolic moieties using NaOH and analyzed in a negative ESI mass spectrometry. Although simple deprotonation works to produce excellent results for many types of lignin compounds, others can undergo extensive in-source fragmentation for certain bond types making structural elucidation more complicated. Herein, we present an alternative method for analyzing lignin model compounds using chloride adduct chemistry. In this study, nine β-O-4 dimers, an (4-Ο-α)(β-Ο-4) trimer, and a (β-O-4)(β-O-4) trimer were synthesized and analyzed using chloride adduct mass spectrometry in the negative mode using NH4Cl as the chloride source. Stable chloride adducted molecular ions were observed for a...

De Jeugdwet is voorlopig de laatste in een reeks wetten waarvan de eerste twee zijn: de Wet op de jeugdhulpverlening (1989) en de Wet op de jeugdzorg (2005). De gemiddelde levensduur van wetten op het terrein van de jeugdhulp was 8,2 jaar. Dat is kort. Momenteel zijn clienten, gemeenten en hulpverleners nog bezig de gevolgen van de invoering van de Jeugdwet in 2015 te verwerken. Toch is er sinds 1 oktober 2016 alweer een evaluatie gaande waarvan de uiteindelijke resultaten in 2018 bekend zullen worden gemaakt en die vermoedelijk niet zonder gevolgen zal blijven. De vraag is of dit niet te veel van het goede is. Het onderzoek wordt uitgevoerd in vijf deelstudies. Een van de studies betreft het wettelijke instrumentarium. Over dit instrumentarium gaat dit artikel. Dit artikel bevat het voorstel om een deel van de oorzaken van de korte levensduur van wetten op het terrein van de jeugdhulp weg te nemen door inhoudelijke doelen uit de Jeugdwet te halen. De wetenschappelijke basis is omstreden en/of de condities waaronder zij werken zijn onvoldoende gespecificeerd. Ook biedt de wet geen ruimte om zonder tijdrovende aanpassingen in te spelen op veranderde maatschappelijke omstandigheden.

The objectives of this study were to evaluate degradation of ergovaline in a tall fescue [ (Schreb.) Darbysh.] seed extract by rumen microbiota ex vivo and to identify specific bacteria capable of ergovaline degradation in vitro. Rumen cell suspensions were prepared by harvesting rumen fluid from fistulated wether goats ( = 3), straining, and differential centrifugation. Suspensions were dispensed into anaerobic tubes with added Trypticase with or without extract (∼10 μg kg ergovaline). Suspensions were incubated for 48 h at 39°C. Samples were collected at 0, 24, and 48 h for ergovaline analysis and enumeration of hyper-ammonia producing (HAB) and tryptophan-utilizing bacteria. Ergovaline values were analyzed by repeated measures using the mixed procedure of SAS. Enumeration data were log transformed for statistical analysis. When suspensions were incubated with extract, 11 to 15% of ergovaline disappearance was observed over 48 h ( = 0.02). After 24 h, suspensions with added extract had 10-fold less HAB than controls ( = 0.04), but treatments were similar by 48 h ( = 1.00). However, after 24 h and 48 h, suspensions with extract had 10-fold more tryptophan-utilizing bacteria ( < 0.01) that were later isolated and identified by their 16S RNA gene sequence as . The isolates and other known rumen pure cultures ( JB1, B159, HD4, B, F, MD1, SR) were evaluated for the ability to degrade ergovaline in vitro. Pure culture cell suspensions were incubated as described above and samples were taken at 0 and 48 h for ergovaline analysis. Data were analyzed using the ANOVA procedure of SAS. All HAB, including the isolates, tested degraded ergovaline (54 to 75%; < 0.05). B14 was also able to degrade ergovaline but to a lesser capacity (12%; < 0.05), but all other bacteria tested did not degrade ergovaline. The results of this study indicate which rumen bacteria may play an important role in ergovaline degradation and that microbiological strategies for controlling their activity could have ramifications for fescue toxicosis and other forms of ergotism in ruminants.

Electrospray ionization mass spectrometry ESI-MS is a powerful technique for the characterization of macromolecules and their noncovalent binding with guest ions. We herein evaluate the feasibility of using ESI-MS as a screening tool for predicting potentiometric selectivities of ionophores. Ion-selective electrodes based on the cyclic peptide, cyclosporin A, were developed, and their potentiometric selectivity pattern was evaluated. Optimized electrodes demonstrated near-Nernstian slopes with micromolar detection limits toward calcium. ESI-MS and ESI-MS/MS were employed to determine the relative association strengths of cyclosporin A with various cations. The observed MS intensities of ion-ionophore complexes correlate favorably with the potentiometric selectivity pattern that was demonstrated by cyclosporin-based electrodes. This correlation was found to hold true for other established ionophores, such as valinomycin and benzo-18-crown-6. Taken together, these experiments demonstrate that mass spectrometry could be used to predict the selectivity patterns of new ionophores for potentiometric and optical ion sensors. Further, this approach could be useful in screening mixtures or libraries of newly-synthesized compounds to identify selective ionophores.

Application of a microfluidic CE device for CZE-MS allows for fast, rapid, and in-depth analysis of large sample sets. The microfluidic CZE-MS device, the 908 Devices ZipChip, involves minimal sample preparation and is ideal for small cation analytes, such as alkaloids. Here, we evaluated the microfluidic device for the analysis of alkaloids from Lobelia cardinalis hairy root cultures. Extracts from wild-type, transgenic, and selected mutant plant cultures were analyzed and data batch processed using the mass spectral processing software MZmine2 and the statistical software Prism 8. In total 139 features were detected as baseline resolved peaks via the MZmine2 software optimized for the electrophoretic separations. Statistically significant differences in the relative abundance of the primary alkaloid lobinaline (C(27)H(34)N(2)), along with several putative “lobinaline-like” molecules were observed utilizing this approach. Additionally, a method for performing both targeted and untargeted tandem mass spectrometry experiments using the microfluidic device was developed and evaluated. Coupling data-processing software with CZE-MS data acquisition has enabled comprehensive metabolomic profiles from plant cell cultures to be constructed within a single working day.

Background Evaluation of clinical and functional results of a new extended depth of focus intraocular lens (EDOF-IOL). Methods Fourteen cataract patients (28 bilateral implantations) were assessed for uncorrected (UDVA) and corrected (CDVA) distance visual acuities; uncorrected (UNVA), distance-corrected (DCNVA) and best corrected (CNVA) near visual acuities; and uncorrected (UIVA) and distance-corrected (DCIVA) intermediate visual acuities - as well as binocular defocus curves. Photopic and mesopic contrast sensitivity was recorded. Reading acuity was evaluated using an electronic reading desk at fixed distances and at the patient’s preferred near and intermediate distances. Visual symptoms were assessed with a halo and glare simulator plus a patient questionnaire which also recorded quality of life. Results Median postoperative monocular UDVA was 0.13logMAR (range − 0.08 to 0.42logMAR), median CDVA was − 0.01logMAR (range − 0.20 to 0.22logMAR), median UIVA at 80 cm was − 0.05logMAR (range − 0.18 to 0.58logMAR) and median UNVA at 40 cm was 0.14logMAR (range − 0.10 to 0.64logMAR). Binocular uncorrected reading acuity was 0.10logMAR at 40 cm and 0.11logMAR at 80 cm. Patients preferred a median intermediate reading distance of 62.8 cm over the predetermined 80 cm, which allowed them to read smaller letter size but did not improve reading acuity. Patients reported a high rate of spectacle independence and satisfaction in everyday life and little to no dysphotopsia. Conclusion The Mini WELL Ready IOL provided good postoperative functional results at far and intermediate distances and improved the visual and reading acuity at reading distance. The lens caused little to no dysphotopsia. Trial registration The study protocol was registered at the German Clinical Trials Register: DRKS00007837 (Registered Date: March 9th, 2015).

Wat kunnen gemeenten als verantwoordelijke voor de jeugdhulp met behulp van hun regie doen om een systeem te creeren waarin de client centraal staat?

We present the case of a 56-year-old woman with moderate myopia and bilateral cataract who had cataract extraction and intraocular lens (IOL) implantation. Due to the patient's desire for spectacle independence, a trifocal IOL with toric correction for astigmatism was implanted. During the follow-up, it became obvious that the implanted IOL had rotated and tilted due to insufficient fixation in the large capsular bag of the myopic eye. An IOL explantation was therefore performed, and the original IOL was exchanged for a bifocal toric IOL with a larger overall diameter. Stable fixation of the IOL in the capsular bag was achieved, and after surgery in the second eye, the patient recovered good bilateral vision. This case illustrates the need for careful selection of IOL diameter and sizing even in patients with moderate myopia due to the potentially larger ocular dimensions in these patients.

The potential to impart surfaces with specific lignin-like properties (i.e. resistance to microbes) remains relatively unexplored due to the lack of well-defined lignin-derived small molecules and corresponding surface functionalization strategies. Here, allyl-modified guaiacyl β-O-4 eugenol (G-eug) lignin-derived dimer is synthesized and attached to mesoporous silica nanoparticles (MSNPs) via click chemistry. The ability of G-eug lignin-dimer functionalized particles to interact with and disrupt synthetic lipid bilayers is compared to that of eugenol, a known natural antimicrobial. Spherical MSNPs (∼150 nm diameter with 4.5 nm pores) were synthesized using surfactant templating. Post-synthesis thiol (SH) attachment was performed using (3-mercaptopropyl) trimethoxysilane and quantified by Ellman’s test. The resultant SH-MSNPs were conjugated with the G-eug dimers or eugenol by a thiol-ene reaction under ultraviolet light in the presence of a photo initiator. From thermogravimetric analysis (TGA), attachment densities of approximately 0.22 mmol eugenol/g particle and 0.13 mmol G-eug dimer/g particle were achieved. The interaction of the functionalized MSNPs with a phospholipid bilayers of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (representing model cell membranes) supported on gold surface was measured using Quartz Crystal Microbalance with Dissipation monitoring (QCM-D). Eugenol-grafted MSNPs in PBS (up to 1 mg/mL) associated with the bilayer and increased the mass adsorbed on the QCM-D sensor. In contrast, MSNPs functionalized with G-eug dimer show qualitatively different behavior, with more uptake and evidence of bilayer disruption at and above a particle concentration of 0.5 mg/mL. These results suggest that bio-inspired materials with conjugated lignin-derived small molecules can serve as a platform for novel antimicrobial coatings and therapeutic carriers.

In this paper, for the first time we develop and benchmark the performance of three down-converting Field Effect Transistor (FE T) resistive mixers at millimeter wave (mm W) frequencies employing $\pmb{0.15-\mu \mathrm{m}}$ enhancement (E)-mode Gallium Arsenide (GaAs) pseudomorphic high electron mobility transistors (pHEMTs), depletion (D)-mode GaAs pHEMTs, and D-mode Gallium Nitride (GaN) pHEMTs. Our experimental results at 27 GHz demonstrate that the E-mode mixer achieves the highest reported input 3rd order intercept point (IIP3) of 37.5dBm at mm W frequencies to the best of our knowledge. Also, operating at the same LO drive up to 20 dBm, the E-mode GaAs mixer impressively out performs both the D-mode GaAs and GaN mixers.

Under the umbrella of the European Network for Light Ion Therapy (ENLIGHT), the project on Union of Light Ion Centers in Europe (ULICE), which was funded by the European Commission (EC/FP7), was carried out from 2009 to 2014. Besides the two pillars on Transnational Access (TNA) and Networking Activities (NA), six work packages formed the pillar on Joint Research Activities (JRA). The current manuscript focuses on the objectives and results achieved within these research work packages: “Clinical Research Infrastructure”, “Biologically Based Expert System for Individualized Patient Allocation”, “Ion Therapy for Intra-Fractional Moving Targets”, “Adaptive Treatment Planning for Ion Radiotherapy”, “Carbon Ion Gantry”, “Common Database and Grid Infrastructures for Improving Access to Research Infrastructures”. The objectives and main achievements are summarized. References to either publications or open access deliverables from the five year project work are given. Overall, carbon ion radiotherapy is still not as mature as photon or proton radiotherapy. Achieved results and open questions are reflected and discussed in the context of the current status of carbon ion therapy and particle and photon beam therapy. Most research topics covered in the ULICE JRA pillar are topical. Future research activities can build upon these ULICE results. Together with the continuous increase in the number of particle therapy centers in the last years ULICE results and proposals may contribute to the further growth of the overall particle therapy field as foreseen with ENLIGHT and new joint initiatives such as the European Particle Therapy Network (EPTN) within the overall radiotherapy community.

BACKGROUND Although elevated serum levels of visinin-like protein 1 (VILIP-1), a neuron-specific calcium sensor protein, are associated with ischaemic stroke, only a single study has evaluated VILIP-1 as a biomarker of traumatic brain injury (TBI). The current proof-of-concept study was designed to determine whether serum VILIP-1 levels increase post-injury in a well-characterized rat unilateral cortical contusion model. METHODS Lateral flow devices (LFDs) rapidly (

Purpose To prospectively compare the diagnostic accuracy of controlled attenuation parameter (CAP) obtained with transient elastography and proton density fat fraction (PDFF) obtained with proton magnetic resonance (MR) spectroscopy with results of liver biopsy in a cohort of adult patients suspected of having nonalcoholic fatty liver disease (NAFLD). Materials and Methods The institutional review board approved this study. Informed consent was obtained from all patients. The authors evaluated 55 patients suspected of having NAFLD (40 men, 15 women). Patients had a median age of 52.3 years (interquartile range [IQR], 43.7-57.6 years) and a median body mass index of 27.8 kg/m2 (IQR, 26.0-33.1 kg/m2). CAP and PDFF measurements were obtained on the same day, within 27 days of biopsy (IQR, 7-44 days). CAP and PDFF were compared between steatosis grades by using the Jonckheere-Terpstra test. Diagnostic accuracies of CAP and PDFF for grading steatosis were assessed with receiver operating characteristic (ROC) analysis. Within-weeks reproducibility (CAP and PDFF) and within-session repeatability were assessed with linear regression analyses, intraclass correlation coefficients, and coefficients of variation. Results Steatosis grades at liver biopsy were distributed as follows: S0, five patients; S1, 24 patients; S2, 17 patients; and S3, nine patients. Both PDFF and CAP helped detect histologically proven steatosis (≥S1), but PDFF showed better diagnostic accuracy than CAP in terms of the area under the ROC curve (0.99 vs 0.77, respectively; P = .0334). PDFF, but not CAP, enabled the grading of steatosis (P < .0001). For within-weeks reproducibility, the intraclass correlation coefficient with PDFF was higher than that with CAP (0.95 vs 0.65, respectively; P = .0015); coefficients of variation were similar (19% vs 11%, P = .55). Within-session repeatability of CAP was good, with a coefficient of variation of 4.5%. Conclusion MR spectroscopy-derived PDFF is superior to CAP in detecting and grading liver steatosis in human NAFLD. © RSNA, 2017 Online supplemental material is available for this article.

Background Laboratory analysis and optical quality assessment of explanted hydrophilic intraocular lenses (IOLs) with clinically significant opacification after posterior lamellar keratoplasty (DMEK and DSAEK). Methods Thirteen opacified IOLs after posterior lamellar keratoplasty, 8 after descemet stripping automated endothelial keratoplasty (DSAEK), 3 after descemet membrane endothelial keratoplasty (DMEK) and 2 after both DSAEK and DMEK were analysed in our laboratory. Analyses included optical bench assessment for optical quality, light microscopy, scanning electron microscopy (SEM) and energy dispersive X-Ray spectroscopy (EDS). Results In all IOLs the opacification was caused by a thin layer of calciumphosphate that had accumulated underneath the anterior optical surface of the IOLs in the area spared by the pupil/anterior capsulorhexis. The calcifications lead to a significant deterioration of the modulation transfer function across all spatial frequencies of the affected IOLs. Conclusions The instillation of exogenous material such as air or gas into the anterior chamber increases the risk for opacification of hydrophilic IOLs irrespective of the manufacturer or the exact composition of the hydrophilic lens material. It is recommended to avoid the use of hydrophilic acrylic IOLs in patients with endothelial dystrophy that will likely require procedures involving the intracameral instillation of air or gas, such as DMEK or DS(A)EK.

A very wideband GaAs MMIC Schottky diode frequency doubler has been designed and tested; it has at least a 10–75 GHz output frequency range with indications it works to 90 GHz. Measured in a test fixture with 1.85 mm connectors over an output frequency of 10–67 GHz, it has 9 to 15 dB conversion loss, −15 to −25 dBc fundamental suppression, −22 to −35 dBc third harmonic suppression, and 15 to 20 dB input return loss, all including interconnect and connector losses. Additionally, calibrated wafer probe tests show it has under 16 dB conversion loss up to 75 GHz, and qualitative wafer probe tests indicates it works up to 90 GHz, in good agreement with simulated results. It is singly balanced with a broadside coupled 4:1 transmission line transformer that provides wideband impedance match and input ground return. This appears to be the widest operating bandwidth reported for a GaAs MMIC diode frequency doubler. A family of frequency doublers and mixers has been designed and built that use this new circuit topology.

In an attempt to mimic white-rot fungi lignin degradation via in vivo Fenton chemistry, solution phase Fenton chemistry (10 g biomass, 176 mmol hydrogen peroxide and 1.25 mmol Fe(2+) in 200 mL of water) was applied to four different biomass feedstocks. An enzymatic saccharification of Fenton pretreated biomass showed an average 212% increase relative to untreated control across all four feedstocks (P0.05, statistically significant). A microbial fermentation of the same Fenton pretreated biomass showed a threefold increase in gas production upon a sequential co-culture with Clostridium thermocellum and Clostridium beijerinckii. These results demonstrate the use of solution phase Fenton chemistry as a viable pretreatment method to make cellulose more bioavailable for microbial biofuel conversion.

Aims: FUsed in sarcoma (FUS) is a multifunctional DNA/RNA-binding protein that possesses diverse roles, such as RNA splicing, RNA transport, DNA repair, translation, and transcription. The network of enzymes and processes regulated by FUS is far from being fully described. In this study, we have focused on the mechanisms of FUS-regulated manganese superoxide dismutase (MnSOD) gene transcription. Results: Here we demonstrate that FUS is a component of the transcription complex that regulates the expression of MnSOD. Overexpression of FUS increased MnSOD expression in a dose-dependent manner and knockdown of FUS by siRNA led to the inhibition of MnSOD gene transcription. Reporter analyses, chromatin immunoprecipitation assay, electrophoretic mobility shift assay, affinity chromatography, and surface plasmon resonance analyses revealed the far upstream region of MnSOD promoter as an important target of FUS-mediated MnSOD transcription and confirmed that FUS binds to the MnSOD promoter and interacts with specificity protein 1 (Sp1). Importantly, overexpression of familial amyotropic lateral sclerosis (fALS)-linked R521G mutant FUS resulted in a significantly reduced level of MnSOD expression and activity, which is consistent with the decline in MnSOD activity observed in fibroblasts from fALS patients with the R521G mutation. R521G-mutant FUS abrogates MnSOD promoter-binding activity and interaction with Sp1. Innovation and Conclusion: This study identifies FUS as playing a critical role in MnSOD gene transcription and reveals a previously unrecognized relationship between MnSOD and mutant FUS in fALS. Antioxid. Redox Signal. 20, 1550–1566.

Production of acetic acid via gas-phase hydration of ketene by water (uncatalyzed and in the presence of an additional water molecule) was theoretically characterized using high-level coupled-cluster methods, followed by a two-dimensional master equation analysis to compute thermal reaction rate constants. The results show that the formation of acetic acid quite likely occurs in high-temperature combustion of biomass, but that the rate of formation should be negligible under ambient atmospheric conditions.

Herein we report the fast pyrolysis of the lignin monomers sinapyl and coniferyl alcohol, as well as mixtures of the two, at 650 °C using pyrolysis–GC/MS. The total ion chromatogram area % of certain marker pyrolysates for each alcohol were summed and used to calculate sinapyl:guaiacyl (S:G) ratios in the mixtures; these ratios were then plotted against the actual molar S:G ratios of the starting material. 13 coniferyl alcohol marker pyrolysates and 9 sinapyl alcohol marker pyrolysates provided acceptable linear correlation, whereas several other marker groups chosen did not correlate to the actual S:G ratio in the starting material. Results indicated that the demethoxylation of sinapyl alcohol and/or its pyrolysates occurs during pyrolysis at 650 °C; however, the amount of demethoxylated products generated is statistically insignificant. Having obtained the pyrolysis profile of the various S:G mixtures, marker pyrolysates for the calculation of the S:G ratio in lignin can be carefully selected according to unique samples. These marker groups can then be calibrated against known S:G ratios to provide analysis of the actual S:G ratio of lignin in biomass. For example, pyrolysates chosen from the pyrolysis of peach pit lignin were calibrated in order to determine the S:G ratio in peach pit lignin. The resulting S:G ratio was similar to that obtained from capillary electrophoresis of the products from KMnO 4 oxidation of the peach pit lignin.

Clostridium thermocellum has the ability to catabolize cellulosic biomass into ethanol, but acetic acid, lactic acid, carbon dioxide, and hydrogen gas (H(2)) are also produced. The effect of hydrogenase inhibitors (H(2), carbon monoxide (CO), and methyl viologen) on product selectivity was investigated. The anticipated effect of these hydrogenase inhibitors was to decrease acetate production. However, shifts to ethanol and lactate production are also observed as a function of cultivation conditions. When the sparge gas of cellobiose-limited chemostat cultures was switched from N(2) to H(2), acetate declined, and ethanol production increased 350%. In resting cell suspensions, lactate increased when H(2) or CO was the inhibitor or when the cells were held at elevated hyperbaric pressure (6.8 atm). In contrast, methyl-viologen-treated resting cells produced twice as much ethanol as the other treatments. The relationship of chemostat physiology to methyl viologen inhibition was revealed by glucose transport experiments, in which methyl viologen decreased the rate of glucose transport by 90%. C. thermocellum produces NAD(+) from NADH by H(2), lactate, and ethanol production. When the hydrogenases were inhibited, the latter two products increased. However, excess substrate availability causes fructose 1,6-diphosphate, the glycolytic intermediate that triggers lactate production, to increase. Compensatory ethanol production was observed when the chemostat fluid dilution rate or methyl viologen decreased substrate transport. This research highlights the complex effects of high concentrations of dissolved gases in fermentation, which are increasingly envisioned in microbial applications of H(2) production for the conversion of synthetic gases to chemicals.