Arabidopsis PROTODERMAL FACTOR2 binds lysophosphatidylcholines and transcriptionally regulates phospholipid metabolism.

Plant homeodomain leucine zipper IV (HD-Zip IV) transcription factors (TFs) contain an evolutionarily conserved steroidogenic acute regulatory protein (StAR)-related lipid transfer (START) domain. While the START domain is required for TF activity, its presumed role as a lipid sensor is not clear. Here we used tandem affinity purification from Arabidopsis cell cultures to demonstrate that PROTODERMAL FACTOR2 (PDF2), a representative member that controls epidermal differentiation, recruits lysophosphatidylcholines (LysoPCs) in a START-dependent manner. Microscale thermophoresis assays confirmed that a missense mutation in a predicted ligand contact site reduces lysophospholipid binding. We additionally found that PDF2 acts as a transcriptional regulator of phospholipid- and phosphate (Pi) starvation-related genes and binds to a palindromic octamer with consensus to a Pi response element. Phospholipid homeostasis and elongation growth were altered in pdf2 mutants according to Pi availability. Cycloheximide chase experiments revealed a role for START in maintaining protein levels, and Pi starvation resulted in enhanced protein destabilization, suggesting a mechanism by which lipid binding controls TF activity. We propose that the START domain serves as a molecular sensor for membrane phospholipid status in the epidermis. Our data provide insights toward understanding how the lipid metabolome integrates Pi availability with gene expression.

The START domain mediates Arabidopsis GLABRA2 dimerization and turnover independently of homeodomain DNA binding.

Class IV homeodomain leucine-zipper transcription factors (HD-Zip IV TFs) are key regulators of epidermal differentiation that are characterized by a DNA-binding HD in conjunction with a lipid-binding domain termed steroidogenic acute regulatory-related lipid transfer (START). Previous work established that the START domain of GLABRA2 (GL2), a HD-Zip IV member from Arabidopsis (Arabidopsis thaliana), is required for TF activity. Here, we addressed the functions and possible interactions of START and the HD in DNA binding, dimerization, and protein turnover. Deletion analysis of the HD and missense mutations of a conserved lysine (K146) resulted in phenotypic defects in leaf trichomes, root hairs, and seed mucilage, similar to those observed for START domain mutants, despite nuclear localization of the respective proteins. In vitro and in vivo experiments demonstrated that while HD mutations impair binding to target DNA, the START domain is dispensable for DNA binding. Vice versa, protein interaction assays revealed impaired GL2 dimerization for multiple alleles of START mutants, but not HD mutants. Using in vivo cycloheximide chase experiments, we provided evidence for the role of START, but not HD, in maintaining protein stability. This work advances our mechanistic understanding of HD-Zip TFs as multidomain regulators of epidermal development in plants.

Prevalence and incidence of hepatitis C infection amongst men who have sex with men in a population-based pre-exposure prophylaxis program in British Columbia, Canada.

BACKGROUND: Men who have sex with men (MSM) are at risk for sexually-transmitted hepatitis C (HCV). Evidence for HCV infection in the context of pre-exposure prophylaxis (PrEP) use in North America is limited. We sought to characterize baseline HCV prevalence and incidence in MSM receiving PrEP in British Columbia (BC), Canada. METHODS: We followed individuals in the BC PrEP program from January 2018 to August 2019. We evaluated baseline prevalence and incident seroconversions (newly positive HCV antibody). A multivariable logistic regression model was performed in MSM for factors associated with HCV prevalence at enrollment, including reported prior sexually transmitted infection (STI), HIV Incidence Risk Index for MSM score, PrEP use because of a partner living with HIV, and location of residence. RESULTS: The median age of the cohort was 33 years, 98.3% male, with 3058 person years (PY) of follow-up. Baseline HCV prevalence was 0.82% (31/3907 MSM enrollees) and HCV incidence (n = 3) was 0.15 per 100 PY (95% confidence interval [CI] 0.03-0.45). In multivariable analysis, initiating PrEP because of a partner living with HIV (adjusted odds ratio [aOR] 5.02; 95% CI 1.87-13.47) and prior STI (aOR 2.34; 95% CI 1.04-5.24) were associated with positive HCV status. CONCLUSIONS: Baseline HCV prevalence and incidence was low amongst MSM in a population-based PrEP program in BC, Canada. HCV was associated with bridging from populations living with HIV and evidence of a reported prior STI as a PrEP indicator condition amongst MSM. PrEP initiation may be an opportunity for linkage to HCV screening and treatment.

Sorption, coagulation, and biodegradation for graywater treatment.

Population growth and climate change are exacerbating water scarcity. Graywater recycling could reduce water demand but it is not commonly practiced because of high treatment costs. Biochar, an emerging low-cost alternative sorbent with potential environmental benefits for graywater treatment, was compared to activated carbon (AC) for removing dissolved organic carbon (DOC) from graywater. The impact of pretreatments (coagulation, biodegradation) were also evaluated. Among five biochars tested, a wood-based biochar was the most effective for graywater treatment, but AC removed more DOC. Sorption resulted in a greater percent removal of ultraviolet (UV) absorbance than DOC or free chlorine demand. Graywater regulations could not be met by sorption alone but could be met with pretreatment before sorption. After biodegradation, irrigation and toilet flushing treatment targets could be achieved with AC doses less than 0.7 g/L, while a biochar dose of about 1 g/L was needed to achieve the irrigation treatment targets. For DOC removal, alum coagulation at a dose of 30 mg/L was a less effective pretreatment than biodegradation. Pretreatment and sorption to decrease turbidity and increase UV transmittance could be effective for the potential use of UV disinfection, thus creating an effective graywater non-potable reuse approach.

Ti-Catalyzed Hydroamination for the Synthesis of Amine-Containing π-Conjugated Materials.

A series of conjugated enamines were prepared by Ti catalyzed anti-Markovnikov hydroamination. The synthetic route is efficient with yields of up to 94 % and the 100 % atom efficiency of the reaction means that these products are easily isolated and purified. Due to the extended conjugated system, the enamine tautomers were observed exclusively in both solid and solution phases, as determined by X-ray crystallography and NMR spectroscopy. These new conjugated molecules, with N incorporated into the backbone, show interesting photophysical properties including photo-luminescent quantum yields of up to 0.26. Notably, through the incorporation of B to give a donor-acceptor π-conjugated system, a redshift of approximately 100 nm is observed for the emission maximum along with the anticipated solvatochromic shifts.

Environmental Comparison of Biochar and Activated Carbon for Tertiary Wastewater Treatment.

Micropollutants in wastewater present environmental and human health challenges. Powdered activated carbon (PAC) can effectively remove organic micropollutants, but PAC production is energy intensive and expensive. Biochar adsorbents can cost less and sequester carbon; however, net benefits depend on biochar production conditions and treatment capabilities. Here, life cycle assessment was used to compare 10 environmental impacts from the production and use of wood biochar, biosolids biochar, and coal-derived PAC to remove sulfamethoxazole from wastewater. Moderate capacity wood biochar had environmental benefits in four categories (smog, global warming, respiratory effects, noncarcinogenics) linked to energy recovery and carbon sequestration, and environmental impacts worse than PAC in two categories (eutrophication, carcinogenics). Low capacity wood biochar had even larger benefits for global warming, respiratory effects, and noncarcinogenics, but exhibited worse impacts than PAC in five categories due to larger biochar dose requirements to reach the treatment objective. Biosolids biochar had the worst relative environmental performance due to energy use for biosolids drying and the need for supplemental adsorbent. Overall, moderate capacity wood biochar is an environmentally superior alternative to coal-based PAC for micropollutant removal from wastewater, and its use can offset a wastewater facility's carbon footprint.

Sources of per- and polyfluoroalkyl substances in an arid, urban, wastewater-dominated watershed.

Per- and polyfluoroalkyl substances (PFAS) enter surface waters from various sources such as wastewater treatment plants, fire-fighting sites, and PFAS-producing and PFAS-using industries. The Las Vegas Wash in Southern Nevada of the United States (U.S.) conveys wastewater effluent from the Las Vegas metropolitan area to Lake Mead, a drinking water source for millions of people in the U.S. Southwest. PFAS have previously been detected in the Las Vegas Wash, but PFAS sources were not identified. In this study, upstream wash tributaries, wastewater treatment effluents, and shallow groundwater wells were sampled in multiple campaigns during dry-weather conditions to investigate possible PFAS sources. Out of 19 PFAS, two short-chain PFAS-perfluoropentanoic acid (48 % of the total molar concentration) and perfluorohexanoic acid (32 %)-comprised the majority of PFAS loading measured in the Las Vegas Wash, followed by perfluorooctanoic acid (9 %). On a mass loading basis, the majority of total measured PFAS (approximately 90 %) and at least 48 % of each specific PFAS in the Las Vegas Wash likely entered via municipal wastewater effluents, of which the main source was likely residential wastewater. One of the drainage areas with a major civilian airport was identified as a potential source of relatively enriched perfluorosulfonic acids to a small wash tributary and shallow groundwater samples. Nonetheless, that tributary contributed at most 15 % of any specific PFAS to the mainstem of the Las Vegas Wash. Total PFAS concentrations were relatively low for the small tributary associated with an urban smaller airport and the lack of flow in the tributary channel immediately downgradient of an Air Force base indicates the smaller airport and base were unlikely significant PFAS sources to the Las Vegas Wash. Overall, this study demonstrated effective PFAS source investigation methodology and the importance of wastewater effluent as a PFAS environmental pathway.

Long-Term Statistical Process Monitoring of an Ultrafiltration Water Treatment Process.

As water treatment technology has improved, the amount of available process data has substantially increased, making real-time, data-driven fault detection a reality. One shortcoming of the fault detection literature is that methods are usually evaluated by comparing their performance on hand-picked, short-term case studies, which yields no insight into long-term performance. In this work, we first evaluate multiple statistical and machine learning approaches for detrending process data. Then, we evaluate the performance of a PCA-based fault detection approach, applied to the detrended data, to monitor influent water quality, filtrate quality, and membrane fouling of an ultrafiltration membrane system for indirect potable reuse. Based on two short case studies, the adaptive lasso detrending method is selected, and the performance of the multivariate approach is evaluated over more than a year. The method is tested for different sets of three critical tuning parameters, and we find that for long-term, autonomous monitoring to be successful, these parameters should be carefully evaluated. However, in comparison with industry standards of simpler, univariate monitoring or daily pressure decay tests, multivariate monitoring produces substantial benefits in long-term testing.

Occurrence of quantifiable and semi-quantifiable poly- and perfluoroalkyl substances in united states wastewater treatment plants.

Both quantifiable and semi-quantifiable poly- and perfluoroalkyl substances (PFAS) were evaluated in the influent, effluent, and biosolids of 38 wastewater treatment plants. PFAS were detected in all streams at all facilities. For the means of the sums of detected, quantifiable PFAS concentrations were 98 ± 28 ng/L, 80 ± 24 ng/L, and 160,000 ± 46,000 ng/kg (dry weight basis) in the influent, effluent, and biosolids (respectively). In the aqueous influent and effluent streams this quantifiable PFAS mass was typically associated with perfluoroalkyl acids (PFAAs). In contrast, quantifiable PFAS in the biosolids were primarily polyfluoroalkyl substances that potentially serve as precursors to the more recalcitrant PFAAs. Results of the total oxidizable precursor (TOP) assay on select influent and effluent samples showed that semi-quantified (or, unidentified) precursors accounted for a substantial portion (21 to 88%) of the fluorine mass compared to that associated with quantified PFAS, and that this fluorine precursor mass was not appreciably transformed to perfluoroalkyl acids within the WWTPs, as influent and effluent precursor concentrations via the TOP assay were statistically identical. Evaluation of semi-quantified PFAS, consistent with results of the TOP assay, showed the presence of several classes of precursors in the influent, effluent, and biosolids; perfluorophosphonic acids (PFPAs) and fluorotelomer phosphate diesters (di-PAPs) occurred in 100 and 92% of biosolid samples, respectively. Analysis of mass flows showed that, for both quantified (on a fluorine mass basis) and semi-quantified PFAS, the majority of PFAS exited WWTPs through the aqueous effluent compared to the biosolids stream. Overall, these results highlight the importance of semi-quantified PFAS precursors in WWTPs, and the need to further understand the impacts of their ultimate fate in the environment.

Using machine learning classification to detect simulated increases of de facto reuse and urban stormwater surges in surface water.

Water quality events such as increases in stormwater or wastewater effluent in drinking water sources pose hazards to drinking water consumers. Stormwater and wastewater effluent enter Lake Mead-an important drinking water source in the southwest USA-via the Las Vegas Wash. Previous studies have applied machine learning and online instruments to detect contamination in water distribution systems. However, alert systems at drinking water intakes would provide more time for corrective action. An array of online instruments measuring pH, conductivity, redox potential, turbidity, temperature, tryptophan-like fluorescence, UV absorbance (UVA254), TOC, and chlorophyll-a was fed raw water directly from Lake Mead. Wastewater effluent, dry weather Las Vegas Wash, and storm-impacted Las Vegas Wash samples were blended into the instrument inlets at known ratios to simulate three types of adverse water quality events. Data preprocessing was conducted to correct for diurnal patterns or instrument drift. Supervised machine learning was conducted using previously published models in R. Ninety-nine models were screened on the raw data. Eight high-performing models were evaluated in-depth and optimized. Weighted k-Nearest Neighbors, Single C5.0 Ruleset, Mixture Discriminant Analysis, and an ensemble of these three models had accuracy over 97% when assigning test set data among three classes (Normal, Event, or Maintenance). The ensemble detected all event types at the earliest timepoint and had one false positive that was not a lag error (i.e., consecutively following a true positive). Omitting Maintenance, the Adaboost model had over 99% test set accuracy and zero false positives that were not lag errors. Data preprocessing was beneficial, but the optimal methods were model-specific. All nine water quality variables were useful for most models, but UVA254 and turbidity were most important.

Chronic Hepatitis B Infection Among Preexposure Prophylaxis Users Enrolled in a Population-Based Program in British Columbia, Canada.

Initiation of human immunodeficiency virus preexposure prophylaxis (PrEP) medications will also treat hepatitis B infection (HBV). The prevalence of chronic HBV was 0.86% (n=41/4760) among enrollees in a provincial PrEP program in British Columbia, Canada. Overall, 46.3% lacked follow-up HBV DNA monitoring, underscoring the need for HBV-related education for PrEP prescribers.

Summation of disinfection by-product CHO cell relative toxicity indices: sampling bias, uncertainty, and a path forward.

The cyto- and genotoxic potencies of disinfection by-products (DBPs) have been evaluated in published literature by measuring the response of exposed Chinese hamster ovary cells. In recent publications, DBP concentrations divided by their individual toxicity indices are summed to predict the relative toxicity of a water sample. We hypothesized that the omission or inclusion of certain DBPs over others is equivalent to statistical sampling bias and may result in biased conclusions. To test this hypothesis, we removed or added actual or simulated DBP measurements to that of published studies which evaluated granular activated carbon as a treatment to reduce the relative toxicity of the effluent. In several examples, it was possible to overturn the conclusions (i.e., activated carbon is detrimental or beneficial in reducing toxicity) by preferentially including specific DBPs. In one example, removing measured haloacetaldehydes caused the predicted cytotoxicity of a treated sample to decrease by up to 47%, reversing the initial conclusion that activated carbon increased the toxicity of the water. We also discuss measurements of statistical error, which are rarely included in publications related to predicted toxicity, but strongly influence the outcomes. Finally, we discuss future research needs in the light of these and other concerns.

Making glue from seeds and gums: Working with plant-based polymers to introduce students to plant biochemistry.

Plants and plant products are key to the survival of life on earth. Despite this fact, the significance of plant biochemistry is often underrepresented in science curricula. We designed an innovative laboratory activity to engage students in learning about the biochemical properties of natural polymers produced by plants. The focus of the hands-on activity is on mucilages and gums, which contain complex polysaccharides that have applications in industry. The 1.5-h activity is organized into three laboratory exercises. It begins with a demonstration of the water absorption property of seed coat mucilage upon hydration of seeds from psyllium, a plant that is grown commercially for mucilage production. The second exercise involves microscopy of a variety of plant seeds stained with ruthenium red dye to visualize pectin polysaccharides of the seed mucilage. Students learn about phenotypic variation among plant species and how the seed coat mucilage is beneficial to keep seeds hydrated during germination. The third exercise highlights an industrial application of plant gums as adhesives. The students prepare edible glue made with gum arabic, a type of plant polymer from the dried exudate of the Acacia plant. This three-part activity has been implemented in conjunction with a Girls Researching Our World (GROW) summer workshop for sixth to eighth graders over a 4-year period. It may be adapted as a laboratory activity for students of all ages, for example, to enhance biochemistry education for high-school students or undergraduate non-majors. © 2019 International Union of Biochemistry and Molecular Biology, 47(4):468-475, 2019.