Dose-response Experiments Research Articles

A bioassay to determine Poa annua responses to indaziflam

Herbicide resistance within Poa annua is widespread in managed turfgrass systems. In 2020, a P. annua collection from a golf course in the southeastern United States was reported to be resistant to indaziflam as well as six other mode-of-action groups. This first report in 2020 suggests that turfgrass managers would benefit from a bioassay to screen other collections with putative indaziflam resistance. A dose-response experiment was conducted with ten concentrations of indaziflam (0, 250, 500, 667, 1000, 1143, 1333, 2000, 4500, and 9000 pM) in Gelrite® culture during 2021 and 2022. An herbicide-susceptible (S1) collection of P. annua, a resistant standard (Site 3A), and a collection with putative resistance to indaziflam (Site 18) were included in this experiment. Petri dishes were filled with 80 mL of Gelrite® (3.75 g L−1) containing technical grade (≥ 98%) indaziflam and rifampicin (1000 µg mL−1). Each plate was sealed with parafilm after placing 15 seeds of a single collection on the Gelrite® surface. At 14 days after seeding (DAS), the length of the radicle (mm) protruding from each seed was recorded with digital calipers. Indaziflam concentrations required to reduce root growth by 70% (EC70) were calculated via non-linear regression. Statistically significant differences were detected among P. annua collections with the EC70 values for the herbicide-susceptible collection measuring 708 pM [95% confidence interval (CI) = 656 to 764 pM] compared to 2130 pM (CI = 1770 to 2644 pM) for Site 3A and 4280 pM (CI = 3464 to 5442) for Site 18. Given that resistant collections exhibited longer root length in the absence of herbicide, confocal microscopy analysis was used to explore differences in root cell count among resistant and susceptible P. annua collections; however, few differences in cell count were detected. Overall, these findings indicate that a discriminatory dose of 708 pM (95% CI = 656 to 764 pM) can be used to differentiate among susceptible and resistant P. annua collections from field sites where poor control is observed following broadcast applications of indaziflam.

Designing microplate layouts using artificial intelligence

Microplates are indispensable in large-scale biomedical experiments but the physical location of samples and controls on the microplate can significantly affect the resulting data and quality metric values. We introduce a new method based on constraint programming for designing microplate layouts that reduces unwanted bias and limits the impact of batch effects after error correction and normalisation. We demonstrate that our method applied to dose-response experiments leads to more accurate regression curves and lower errors when estimating IC50/EC50, and for drug screening leads to increased precision, when compared to random layouts. It also reduces the risk of inflated scores from common microplate quality assessment metrics such as Z′ factor and SSMD. We make our method available via a suite of tools (PLAID) including a reference constraint model, a web application, and Python notebooks to evaluate and compare designs when planning microplate experiments.

Potential bio-herbicide from Streptomyces sp. KR0005 to control weeds in a horticultural field

ABSTRACT Herbicidal active metabolites obtained from actinomycetes provide valuable sources in the search for new candidates. They might be striking alternatives to synthetic herbicides. In the present study, actinobacteria strains were isolated from soil samples and their herbicidal activities were screened. In the screening, Streptomyces sp. strain KR0005 broth culture exhibited the highest phytotoxic activity. Thus, it was selected for further studies. The herbicidal active compound purified from this selected strain was identified as cycloheximide (MW, 281.; C15H23NO4) based on 1H-NMR, 13C-NMR, 2D-NMR, and high-resolution electrospray ionisation mass spectrometry analyses. Dose–response experiments revealed that the GR50 value of Digitaria ciliaris was 1.37 g L−1 to Streptomyces sp. strain KR0005 culture filtrate. In general, the inclusion of v/v 0.1% of LES270 (Alcohols, (C12-14), ethoxylated, mono ethers with sulphuric acid, sodium salts) and SF90 (Alcohols, C12-14-secondary, ethoxylated) surfactants resulted in a 14.3% decrease and a 24.1% increase in herbicidal activity, respectively, compared with Tween-20 (Polyoxyethylene sorbitan monolaurate). Mode of action experiments revealed that the KR0005 culture filtrate acted as both photosynthesis and cell membrane integrity inhibitors in a dose-dependent manner. However, further studies such as mutation and/or specific culture conditions are needed to increase the herbicidal activity of this selected strain.

Cathepsin-L Secreted by High-Quality Bovine Embryos Exerts an Embryotrophic Effect In Vitro.

While human in vitro embryo production is generally performed individually, animal models have shown that culturing embryos in groups improves blastocyst yield and quality. Paracrine embryotrophins could be responsible for this improved embryo development, but their identity remains largely unknown. We hypothesize that supplementation of embryotrophic proteins to a culture medium could be the key to improve individual embryo production. In this study, proteomics screening of culture media conditioned by bovine embryos revealed cathepsin-L as being secreted by both excellent- and good-quality embryos, while being absent in the medium conditioned by poor-quality embryos. The embryotrophic role of cathepsin-L was explored in vitro, whereby bovine zygotes were cultured individually for 8 days with or without cathepsin-L. Preliminary dose-response experiments pointed out 100 ng/mL as the optimal concentration of cathepsin-L in embryo culture medium. Supplementation of cathepsin-L to individual culture systems significantly improved blastocyst development and quality in terms of blastocoel formation at day 7, and the hatching ratio and apoptotic cell ratio at day 8, compared to the control. Taken together, cathepsin-L acts as an important embryotrophin by increasing embryo quality, and regulating blastulation and hatching in bovine in vitro embryo production.

Temperature influences glyphosate efficacy on glyphosate-resistant and -susceptible goosegrass (Eleusine indica).

Glyphosate has been widely used to control Eleusine indica and other weeds in South China for many years. Among the most troublesome weeds in South China, E. indica can remain alive all year round. However, the influence of temperature on glyphosate efficacy on E. indica, especially under days with fluctuating temperature, is unknown. This study evaluated the influence of two temperature regimes on glyphosate efficacy on glyphosate-resistant (R) and -susceptible (S) E. indica biotypes. Plants of the R and S biotypes were cultivated under two temperature regimes (high: 30°C/20°C day/night; low: 20°C/15°C day/night). Dose-response experiments showed improved efficacy of glyphosate at the low temperature compared with that at the high temperature for both biotypes. Based on the LD50 values, the R biotype was 8.9 times more resistant to glyphosate than the S biotype at the high temperature; however, the resistance index (R/S) decreased to 3.1 at the low temperature. At 4 days after glyphosate application, shikimic acid accumulation was greater at the low temperature than at the high temperature in plants of both biotypes, and the increase was higher in plants of the R biotype than in the S biotype. At a sublethal glyphosate dose (R: 400 g ai ha-1; S: 200 g ai ha-1), plants grown at the low temperature showed a strong decrease in leaf chlorophyll content and Fv/Fm value compared with those of plants grown at the high temperature and the untreated control. At 3 days after treatment, glyphosate absorption was similar between biotypes at the high temperature, but absorption decreased to 64.9% and 53.1% at the low temperature for the R and S biotypes, respectively. For both biotypes, glyphosate translocation from the leaf to the remainder of the plant was reduced at the low temperature compared with that at the high temperature. No differences in glyphosate translocation were observed between biotypes within each temperature regime. This is the first report on the effect of temperature on glyphosate efficacy on E. indica, and provides important insights for glyphosate application and resistance management.

Solid-Supported Membrane (SSM)-Based Electrophysiology Assays Using Surface Electrogenic Event Reader Technology (SURFE²R) in Early Drug Discovery.

This article presents detailed descriptions of procedures and troubleshooting tips for solid-supported membrane (SSM)-based electrophysiology assays (SURFE²R) to measure electrogenic solute carrier transporter proteins (SLCs) and assess the effects of compounds that modulate their activity. SURFE²R allows the use of the standard 96-well format, making it an ideal platform for tertiary assays in a drug-discovery campaign. The assays are performed with cell-line-derived membrane fractions or proteoliposomes containing the transporter of interest. Three main protocols are described for the isolation of membrane fractions from cell culture and the generation of proteoliposomes containing the transporter of interest. Additionally, detailed protocols for SURFE²R single concentration and dose-response experiments are included to measure the potencies of test compounds in stimulating or inhibiting transporter function (EC50 or IC50 values, respectively) and kinetic functional assays to calculate apparent affinity (kM ) and maximal velocity (Vmax ) of substrate uptake. © 2023 Sanofi. Current Protocols published by Wiley Periodicals LLC. PROTOCOL GROUP 1: Sample preparation for SSM-based electrophysiology assays Support Protocol 1: Production of cell batches Support Protocol 2: Simple isolation of cell membranes Alternate Protocol 1: Isolation of cell membranes with sucrose gradient pre-purification Support Protocol 3: Production and isolation of liposomes Support Protocol 4: Preparation of sensor with isolated cell membranes Alternate Protocol 2: Preparation of sensor with isolated proteoliposomes PROTOCOL GROUP 2: Determination of assay parameters for SSM-based electrophysiology assay Support Protocol 5: Assay with stable buffer Alternate Protocol 3: Assay with ion gradient Support Protocol 6: Determination of membrane/liposome concentration Support Protocol 7: Determination of substrate dependency kM PROTOCOL GROUP 3: Determination of advanced assay parameters for SSM-based electrophysiology assays Support Protocol 8: Assessment of ion concentration dependency Support Protocol 9: Assessment of pH dependency Support Protocol 10: Assessment of DMSO dependency Support Protocol 11: Assessment of signal stability with multiple activations PROTOCOL GROUP 4: Compound testing through SSM-based electrophysiology assays using SURFE²R apparatus Support Protocol 12: Assessment of signal specificity of a published inhibitor or unknown compound(s) Support Protocol 13: Compound wash-out Support Protocol 14: Statistical analysis.

Non-Target Site Resistance in Summer-Emerging Lolium rigidum and the Effect of Alternative Herbicides

Herbicide resistance is an important weed management issue. Glyphosate is the most dominant herbicide, which controls a broad spectrum of weeds, including grasses such as Lolium rigidum. Lolium rigidum is a major weed of winter crops in Australia that is developing glyphosate resistance in increasing numbers of populations and has been observed growing throughout summer in recent years. Three L. rigidum populations, one summer-emerging glyphosate-resistant (GR), one winter-emerging glyphosate-susceptible (GS), and one winter-emerging population with unknown resistance status (CC04), were analyzed for target-site resistance to glyphosate after confirming their resistance or susceptibility to glyphosate in a dose–response experiment. Population GR was obtained from plants that emerged in summer and contained plants that survived all rates of glyphosate applied (0 to 4560 g a.e. ha−1). It was found to be 6.1 and 4.4 times more resistant than population GS across two experiments. Population CC04 was identified as susceptible. Plants from each population were analyzed for the presence of target-site mutations in the conserved region of the EPSPS gene; however, no mutations were identified that could cause resistance, suggesting non-target-site resistance in population GR. The effectiveness of alternative herbicides was also analyzed for each population. Paraquat was the most effective herbicide, with 0% survival across all populations. The ACCase-inhibiting herbicide clethodim was also highly effective (0 to 8% survival across populations). Other ACCase-inhibiting herbicides, propaquizafop, haloxyfop, and pinoxaden, were effective at controlling the two susceptible populations, CC04 and GS (0 to 36% and 0 to 20% survival, respectively), but were only moderately effective for controlling GR (28 to 51% survival). Paraquat and clethodim may be alternative options for controlling GR populations of L. rigidum and could be effective for use in management programs to slow the development of GR populations.

Behavioural and Electrophysiological Response of Anastrepha fraterculus (Diptera: Tephritidae) to a γ-Lactone Synthetic Semiochemical.

Attractants are a powerful tool for pest management. The lack of specific attractants for the South American fruit fly, Anastrepha fraterculus, a complex of cryptic species of great economic importance in South America, makes it difficult to monitor the pest in the field. The γ-lactone male sex and aggregation pheromones of several Anastrepha species, naturally released in a 7:3 epianastrephin to anastrephin ratio, and a structurally related naturally occurring γ-lactone ((±)-trans-tetrahydroactinidiolide) with gem-dimethyl groups (dimethyl) at C(4), were evaluated as potential attractants of this species. Different age and mating conditions of A. fraterculus males and females were evaluated during electroantennography (EAG) and field cage experiments in which polymeric lures were deployed to contain 100 mg of attractant. Epianastrephin and dimethyl were EAG+ for all fly conditions, with epianastrephin eliciting the highest response for both sexes and immature flies showing greater responsiveness than mature flies. In the field cage experiments, immature flies were only attracted to leks; virgin females were attracted to leks, dimethyl, and both epianastrephin-anastrephin formulations (95 and 70 wt.% epianastrephin); mature-mated males were attracted to leks, dimethyl and 70 wt.% epianastrephin; and mature-mated females were only attracted to leks. Our bioassays showed a promising performance of the analog dimethyl since it elicited the same response as epianastrephin, requires fewer steps to synthesize, and contains one less chiral center than the natural pheromones. The attraction to leks was recorded for all mating conditions and ages of flies and suggests that air-borne volatiles of calling males contain cues that could act as sensory traps. The addition of any of these compounds in the synthetic attractants may result in a greater attraction and thus deserves further evaluation. Dose-response experiments will provide additional information to move a step forward and validate the results obtained in open-field conditions.

AI4DR: Development and implementation of an annotation system for high-throughput dose-response experiments

One of the common strategies to identify novel chemical matter in drug discovery consists in performing a High Throughput Screening (HTS). However, the large amount of data generated at the dose-response (DR) step of an HTS campaign requires a careful analysis to detect artifacts and correct erroneous datapoints before validating the experiments. This step which requires to review each DR experiment can be time consuming and prone to human errors or inconsistencies. AI4DR is a system that has been developed for the classification of DR curves based on a Convolutional Neural Network (CNN) acting on normalized images of the DR curves. AI4DR allows the annotation in minutes of thousands of curves among 14 categories to help the High Throughput Screening biologists in their analyses. Several categories are associated with active and inactive compounds, other categories correspond to features of interest such as the presence of noise, a weaker effect at high doses, or a suspiciously weak or strong slope at the inflexion point of the DR curves of actives. The classifier has been trained on an algorithmically generated dataset curated and refined by experts, tested using real screening campaigns and improved using thousands of annotations by experts. The solution is deployed using a MLFlow model server interfaced with the Genedata Screener data analysis software used by the end users. AI4DR improves the consistency, the robustness, and the speed of HTS data analysis as well as reducing the human effort to identify faster new medicines for patients.

Efficacy of Beauveria bassiana against adults of Prostephanus truncatus (Horn), Sitophilus zeamais Motschulsky and Teretrius nigrescens Lewis in stored maize

The larger grain borer, Prostephanus truncatus (Horn) and the maize weevil, Sitophilus zeamais Motschulsky continue to cause tremendous losses to stored maize. Research in the UK has identified Beauveria bassiana, IMI 389521 as a suitable control agent for grain storage pests in the UK. The pathogenicity of B. bassiana, IMI 389521 was evaluated against adult P. truncatus, S. zeamais and Teretrius nigrescens in Ghana. Fifty adults of each insect species were treated with 0.5 g dry conidia powder of this isolate at 8.65 × 108 conidia/g for 1 minute and mortality recorded daily for 14 days. The results indicated that B. bassiana, is pathogenic against P. truncatus and S. zeamais, inducing over 90% mortality by day 7. Teretrius nigrescens was, however less susceptible to the fungus with 30% mortality. To determine the most effective concentration of B. bassiana for the control of P. truncatus, a laboratory dose response experiment using four concentrations of B. bassiana (108–1011 cfu/kg maize) was also conducted. Maize grains (250 g) in separate jars were treated with the four concentrations of the product. Fifty adults of P. truncatus were placed into the jars containing the treated maize and mortality was assessed weekly for 3 weeks. The most effective dose was 1010 cfu/kg maize, which resulted in 96% and 100% mortality of P. truncatus after 2 and 3 weeks, respectively. This study shows that B. bassiana could effectively be integrated into bio-control programme of these two key pests of maize in Ghana after further field trials.

Role of angiotensin receptor blockers in the context of Alzheimer's disease.

As the world's population ages, the prevalence of age-related neurological disorders such as Alzheimer's disease (AD) is increasing. There is currently no treatment for Alzheimer's disease, and the few approved medications have a low success rate in lowering symptoms. As a result, several attempts are underway worldwide to identify new targets for the therapy of Alzheimer's disease. In preclinical studies of Alzheimer's disease, it was recently found that inhibition of angiotensin-converting enzyme (ACE) and blocking of the angiotensin II receptors reduce symptoms of neurodegeneration, Aβ plaque development, and tau hyperphosphorylation. Angiotensin II type I (AT1) blockers, such as telmisartan, candesartan, valsartan, and others, have a wide safety margin and are commonly used to treat hypertension. Renal and cardiovascular failures are reduced due to their vascular protective actions. Inhibition of AT1 receptors in the brain has a neuroprotective impact in humans, reducing the risk of stroke, increasing cognition, and slowing the progression of Alzheimer's disease. The review focuses on the mechanisms via which AT1 blockers may act beneficially in Alzheimer's disease. Although their effect is evident in preclinical studies, clinical trials, on the other hand, are in short supply to validate the strategy. More dose-response experiments with possible AT1 blockers and brain-targeted administration will be needed in the future.

The Contractile Response to Oxytocin in Non-pregnant Rat Uteri Is Modified After the First Pregnancy

During pregnancy, the uterus undergoes several modifications under the influence of hormonal and mechanical stimuli. We hypothesize that while most of these modifications are reverted during involution, some of the physiological properties of the uterus are permanently altered. To investigate this hypothesis, we conducted motility experiments to evaluate the contractility response of uterine tissue samples from non-pregnant virgin and proven breeder female rats to oxytocin (10−10 to 10−5 M). We found that the virgin tissue contracts more robustly than proven breeder tissue in the absence of oxytocin, yet with oxytocin, proven breeder samples displayed a significantly higher increase in activity. These results could depend on a more elevated expression of oxytocin receptor and/or on an alteration in the intracellular pathways affected by the activation of the oxytocin receptors. Here, we explored the impact of some structures involved in the management of intracellular calcium on the dose response to oxytocin recorded from virgin and proven breeder uterine strips. Specifically, we replicated the dose response experiments in low extracellular calcium (10 μM), in the presence of the intracellular calcium channel blocker ruthenium red (10 μM), and in the presence of the sarcoplasmic-endoplasmic reticulum calcium ATP-ase pump inhibitor, cyclopiazonic acid (10 μM). The results of these experiments suggest that also the expression of proteins that control intracellular calcium availability is affected by the experience of pregnancy. Molecular biology experiments will give us more detail on the magnitude of these expression changes.

Development of a double-antibody sandwich ELISA for rapidly quantitative detection of residual non-structural proteins in inactivated foot-and-mouth disease virus vaccines

Foot-and-mouth disease (FMD) is a highly contagious and economically devastating disease of cloven-hoofed animals. Vaccination and surveillance against non-structure protein (NSP) are the most efficacious and cost-effective strategy to control this disease. Therefore, vaccine purity control is vital for successful prevention. Currently, vaccine purity is tested by an in-vivo test that recommended in the World Organization for Animal Health (WOAH), but it is time consuming and costly. Herein, we develop a double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for quantitative detection of residual NSPs in inactivated FMD virus (FMDV) vaccines. In this assay, the monoclonal antibody 3A24 was selected as capture antibody and biotinylated 3B4B1 (Biotin-3B4B1) as detection antibody. A standard curve was developed using the NSP 3AB concentration versus OD value with the linear range of concentration of 2.5–160 ng/mL. The lowest limit of detection was 2.5 ng/mL. In addition, we determined 2.5 ng/mL of NSP as an acceptable threshold value of FMD vaccine purity using a dose-response experiment in cattle. The DAS-ELISA combined with the threshold value of FMD vaccine purity could provide a quick and simple tool for evaluation the antigenic purity of FMD vaccine during the manufacturing process.

Using Single-Species and Whole Community Stream Mesocosm Exposures for Identifying Major Ion Effects in Doses Mimicking Resource Extraction Wastewaters.

Wastewaters and leachates from various inland resource extraction activities contain high ionic concentrations and differ in ionic composition, which complicates the understanding and effective management of their relative risks to stream ecosystems. To this end, we conducted a stream mesocosm dose-response experiment using two dosing recipes prepared from industrial salts. One recipe was designed to generally reflect the major ion composition of deep well brines (DWB) produced from gas wells (primarily Na+, Ca2+, and Cl-) and the other, the major ion composition of mountaintop mining (MTM) leachates from coal extraction operations (using salts dissociating to Ca2+, Mg2+, Na+, SO42- and HCO3-)-both sources being extensive in the Central Appalachians of the USA. The recipes were dosed at environmentally relevant nominal concentrations of total dissolved solids (TDS) spanning 100 to 2000 mg/L for 43 d under continuous flow-through conditions. The colonizing native algal periphyton and benthic invertebrates comprising the mesocosm ecology were assessed with response sensitivity distributions (RSDs) and hazard concentrations (HCs) at the taxa, community (as assemblages), and system (as primary and secondary production) levels. Single-species toxicity tests were run with the same recipes. Dosing the MTM recipe resulted in a significant loss of secondary production and invertebrate taxa assemblages that diverged from the control at all concentrations tested. Comparatively, intermediate doses of the DWB recipe had little consequence or increased secondary production (for emergence only) and had assemblages less different from the control. Only the highest dose of the DWB recipe had a negative impact on certain ecologies. The MTM recipe appeared more toxic, but overall, for both types of resource extraction wastewaters, the mesocosm responses suggested significant changes in stream ecology would not be expected for specific conductivity below 300 µS/cm, a published aquatic life benchmark suggested for the region.

Oral administration of VDAC1-derived small molecule peptides increases circulating testosterone levels in male rats.

Cholesterol is the precursor of all steroid hormones, and the entry of cholesterol into the mitochondria is the rate-limiting step of steroidogenesis. Voltage-dependent anion channel (VDAC1) is an outer mitochondrial protein part of a multiprotein complex that imports cholesterol. We previously reported that intratesticular administration of a 25 amino acid peptide blocking the interaction between 14-3-3ϵ with VDAC1 increased circulating levels of testosterone. This fusion peptide was composed of a HIV-1 transactivator of transcription (TAT) protein transduction domain cell-penetrating peptide, a glycine linker, and amino acids 159-172 of VDAC1 (TV159-172). Here, we describe the development of a family of small molecules that increase circulating testosterone levels after an oral administration. We first characterized an animal model where TV159-172 was delivered subcutaneously. This subcutaneous model allowed us to study the interactions between TV159-172 and the hypothalamus-pituitary-gonadal axis (HPG) and identify the biologically active core of TV159-172. The core consisted of the tetrapeptide RVTQ, which we used as a platform to design synthetic peptide derivatives that can be administered orally. We developed a second animal model to test various derivatives of RVTQ and found 11 active compounds. Dose-response experiments identified 4 synthetic peptides that robustly increased androgen levels in a specific manner. We selected RdVTQ as the leading VDAC1-core derivative and profiled the response across the lifespan of Brown-Norway rats. In summary, we present the development of a new class of therapeutics that act within the HPG axis to increase testosterone levels specifically. This new class of small molecules self-regulates, preventing abuse.

Effect of 3-nitrooxypropanol on enteric methane emissions of feedlot cattle fed a tempered barley-based diet with canola oil.

A dose-response experiment was designed to examine the effect of 3-nitrooxypropanol (3-NOP) on methane (CH4) emissions, rumen function and performance of feedlot cattle fed a tempered barley-based diet with canola oil. Twenty Angus steers of initial body weight (BW) of 356 ± 14.4kg were allocated in a randomized complete block design. Initial BW was used as the blocking criterion. Cattle were housed in individual indoor pens for 112 d, including the first 21 d of adaptation followed by a 90-d finishing period when five different 3-NOP inclusion rates were compared: 0mg/kg DM (control), 50mg/kg DM, 75mg/kg DM, 100mg/kg DM and 125mg/kg DM. Daily CH4 production was measured on d 7 (last day of starter diet), 14 (last day of the first intermediate diet), and 21 (last day of the second intermediate diet) of the adaptation period and on d 28, 49, 70, 91 and 112 of the finisher period using open circuit respiration chambers. Rumen digesta samples were collected from each steer on the day prior to chamber measurement post-feeding, and pre-feeding on the day after the chamber measurement, for determination of rumen volatile fatty acids (VFA), ammonium-N, protozoa enumeration, pH and reduction potential. Dry matter intake (DMI) was recorded daily and BW weekly. Data were analyzed in a mixed model including period, 3-NOP dose and their interaction as fixed effects, and block as a random effect. Our results demonstrated both a linear and quadratic (decreasing rate of change) effect on CH4 production (g/d) and CH4 yield (g/kg DMI) as 3-NOP dose increased (P < 0.01). The achieved mitigation for CH4 yield in our study ranged from approximately 65.5% up to 87.6% relative to control steers fed a finishing feedlot diet. Our results revealed that 3-NOP dose did not alter rumen fermentation parameters such as ammonium-N, volatile fatty acids (VFA) concentration nor VFA molar proportions. Although this experimental design was not focused on the effect of 3-NOP dose on feedlot performance, no negative effects of any 3-NOP dose were detected on animal production parameters. Ultimately, the knowledge on the CH4 suppression pattern of 3-NOP may facilitate sustainable pathways for the feedlot industry to lower its carbon footprint.

Intervention study to inhibit the glucose metabolic remodeling in hepatocytes induced by high-selenium in vitro

To observe the effect of exogenous serine or glycine on the synthesis of selenoprotein and endogenous serine and the expression of metabolic enzymes in hepatocytes cultured with high-selenium in vitro and its dose-response relationship. The experiment was divided into two parts, namely a inhibition experiment and a dose-response experiment, using L02 cells as the intervention target. In the inhibition experiment, the blank control group, high-Se(SeMet) group, serine intervention group and high-Se+serine intervention group were set up. Both SeMet and serine were given at a level of 0.05 μmol/L, and the blank control group was given the same volumes of saline. In the dose-response experiment, the concentration of SeMet was 0.05 μmol/L, and the intervention concentration gradients of serine or glycine were 0, 0.05, 0.1, 0.5, 1, 5, 10, 50, 100 and 500 μmol/L. The expression of phosphoglycerate dehydrogenase(PHGDH)、serine hydroxymethyltransferase 1(SHMT1)、methylenetetrahydrofolate reductase(MTHFR)、selenoprotein P(SELENOP) and glutathione peroxidase 1(GPX1)was detected by Western Blot(WB). (1)In the inhibition experiment, compared with the blank control group, the expression of selenium proteins(GPX1 and SELENOP) in L02 cells of the other three groups were significantly increased(P&lt;0.05). Compared with the high expression of PHGDH in L02 cells of high-Se group, the expressions of PHGDH, SHMT1 and MTHFR in high-Se + serine group were significantly decreased(P&lt;0.05). (2) In the dose-response experiment, the expression of PHGDH enzyme in L02 cells gradually decreased with the increase of the concentration of exogenous serine or glycine, showing an obvious dose-dependent effect. In contrast, none of the other metabolic enzymes(SHMT1 and MTHFR) showed similar trends in protein expression. The upregulated expression of PHGDH, the key enzyme in the de novo synthesis pathway of serine in hepatocytes cultured with high-selenium can be inhibited feedback by exogenous serine or endogenous serine transformed from exogenous glycine directly.

Sensitivity of herbicide-resistant rigid ryegrass (Lolium rigidum) populations to cinmethylin, a new herbicide site of action

AbstractRigid ryegrass (Lolium rigidum Gaudin) is the most problematic weed in Australia, with evolved resistance to multiple herbicide sites of action. Selection pressure by cinmethylin (Group 30, a fatty-acid thioesterase inhibitor) has been limited, because few populations have been exposed to the herbicide since its introduction in 2019. In this study, we examined the sensitivity of L. rigidum populations to this new herbicide. From a screening of almost 500 field populations in 2020, 28 potentially resistant populations were further investigated in a dose–response experiment. Seedlings from five populations surviving treatments of 250 or 375 g ai ha−1 cinmethylin were grown to maturity and seeds were harvested. The level of resistance found among the five putative-resistant parental populations of L. rigidum was negligible. In one population, one round of selection with cinmethylin resulted in a 2-fold increase in the lethal dose causing 50% mortality in the progeny population, although this dose was still only one-sixth of the recommended field rate of cinmethylin. Having a unique site of action, cinmethylin is a viable preemergence herbicide option to control existing multiple-resistance populations of L. rigidum. Comprehensive field monitoring and recurrent selection studies under controlled environmental conditions are needed to better ascertain the risk of L. rigidum evolving a high level of resistance to cinmethylin, although current data suggest that this risk is relatively low.

Fate and PPARγ and STATs-driven effects of the mitochondrial complex I inhibitor tebufenpyrad in liver cells revealed with multi-omics.

The biological effects of the pesticide and mitochondrial complex I inhibitor tebufenpyrad (TEBU) on liver cells were investigated by combining proteomics and metabolomics. Both cell culture media and cellular lysates were analyzed in dose-response and kinetic experiments on the HepaRG cell line. Responses were compared with those obtained on primary human and rat hepatocytes. A multitude of phase I and II metabolites (>80) mainly common to HepaRG cells and primary hepatocytes and an increase in metabolization enzymes were observed. Synthesis of mitochondrion and oxidative phosphorylation complex constituents, fatty acid oxidation, and cellular uptake of lipids were induced to compensate for complex I inhibition and the decrease in ATP intracellular contents caused by TEBU. Secretion of the 20S circulating proteasome and overall inhibition of acute inflammation followed by IL-6 secretion in later stages were observed in HepaRG cells. These effects were associated with a decrease in STAT1 and STAT3 transcription factor abundances, but with different kinetics. Based on identified TEBU targets, docking experiments, and nuclear receptor reporter assays, we concluded that liver cell response to TEBU is mediated by its interaction with the PPARγ transcription factor.

Case Report of Avena sterilis subsp. sterilis ACCase Herbicide Resistance in Southern Spain

Wild oats are worldwide grassy weeds that cause substantial yield losses, particularly in winter cereal crops. In addition, wild oat herbicide resistant cases have increased; indeed, up to 52 cases have been registered. Despite this, no wild oat herbicide resistant cases have been described in Spain, where farmers and technicians have reported poor herbicide efficacy in sterile oats (Avena sterilis subsp. sterilis L.). A dose-response experiment was conducted comparing the behavior of two populations of A. sterilis from southern Spain to a susceptible population. These populations were collected from two commercial farms where a low efficacy of chemical control had been described. Clodinafop-propargyl and Pinoxaden were tested as active ingredients in the dose-response experiment. Additionally, an alternative herbicide, which consisted of a mixture of Mesosulfuron-methyl and Propoxycarbazone-Na, was also tested at a field dose. The two populations of A. sterilis studied provided a resistant factor higher than 10 for Clodinafop-propargyl and higher than 4 for Pinoxaden. A total control was achieved for plants treated with Mesosulfuron-methyl and Propoxycarbazone-Na.