Treatment of chickens with lactobacilli prior to challenge with Clostridium perfringens modifies innate responses and gut morphology.

Necrotic enteritis caused by Clostridium perfringens (CP), is a common enteric disease of poultry that has been previously controlled by in-feed antibiotics. However, due to the rapid emergence of antimicrobial resistance, alternatives to antibiotics such as probiotics have received considerable attention because of their immunomodulatory and intestinal health benefits. The present study investigated the effects of probiotic lactobacilli on gut histomorphology and intestinal innate responses in chickens. Day-old male broiler chickens were treated with 1 × 107 or 1 × 108 colony-forming units (CFU) of a lactobacilli cocktail on days 1, 7, 14, and 20 post-hatch, while control groups were not treated with lactobacilli. On day 21, birds in all groups (except the negative control) were challenged with 3 × 108 CFU of CP for 3 days. Intestinal tissue samples were collected before and after the CP challenge to assess gene expression and for histomorphological analysis. Lactobacilli treatment at a dose of 1 × 108 CFU conferred partial protection against NE by lowering lesion scores, increasing villus height in the ileum and reducing crypt depth in the jejunum. In addition, 1 × 108 CFU of lactobacilli enhanced the expression of Toll-like receptor (TLR) 2, interferon-gamma (IFN-γ), interleukin (IL)-10, IL-12, and IL-13 in both the jejunum and ileum at different timepoints and subsequently decreased the expression of transforming growth factor beta (TGF-ß) and IL-1ß post-CP challenge. In conclusion, the results indicate that treatment with lactobacilli mitigated NE in a dose-dependent manner via improvement of intestinal morphology and modulation of innate immune response in chickens.

In vitro generated antibodies guide thermostable ADDomer nanoparticle design for nasal vaccination and passive immunization against SARS-CoV-2.

Background: Due to COVID-19, pandemic preparedness emerges as a key imperative, necessitating new approaches to accelerate development of reagents against infectious pathogens. Methods: Here, we developed an integrated approach combining synthetic, computational and structural methods with in vitro antibody selection and in vivo immunization to design, produce and validate nature-inspired nanoparticle-based reagents against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Results: Our approach resulted in two innovations: (i) a thermostable nasal vaccine called ADDoCoV, displaying multiple copies of a SARS-CoV-2 receptor binding motif derived epitope and (ii) a multivalent nanoparticle superbinder, called Gigabody, against SARS-CoV-2 including immune-evasive variants of concern (VOCs). In vitro generated neutralizing nanobodies and electron cryo-microscopy established authenticity and accessibility of epitopes displayed by ADDoCoV. Gigabody comprising multimerized nanobodies prevented SARS-CoV-2 virion attachment with picomolar EC50. Vaccinating mice resulted in antibodies cross-reacting with VOCs including Delta and Omicron. Conclusion: Our study elucidates Adenovirus-derived dodecamer (ADDomer)-based nanoparticles for use in active and passive immunization and provides a blueprint for crafting reagents to combat respiratory viral infections.

Thoracic to lumbar vertebral column length and length ratios in miniature dachshunds with and without thoracolumbar intervertebral disc extrusion.

BACKGROUND: The chondrodystrophic body type predisposes miniature dachshunds to thoracolumbar intervertebral disc extrusion (IVDE). However, the relationship between thoracolumbar IVDE and the relative lengths of the thoracic and lumbar vertebral columns has not yet been evaluated. METHODS: This prospective multicentre study included 151 miniature dachshunds with (n = 47) and without (n = 104) thoracolumbar IVDE. All dogs had their thoracic and lumbar vertebral columns measured with a tape measure. Detailed descriptions were provided to facilitate consistent measurement. A thoracic to lumbar vertebral column ratio was calculated. Thoracolumbar IVDE was confirmed by magnetic resonance imaging or computed tomography. RESULTS: The thoracic to lumbar vertebral column length ratio and absolute thoracic vertebral column length were significantly smaller in miniature dachshunds with IVDE than in those without IVDE (p < 0.0001 for both). There were no significant differences in lumbar vertebral column length, age, sex or neuter status between the two groups. LIMITATIONS: The dogs without IVDE did not undergo a neurological examination and the thoracic and lumbar vertebral column measurements were not validated. CONCLUSIONS: The relative lengths of the thoracic and lumbar vertebral column segments could contribute to the development of thoracolumbar IVDE in miniature dachshunds. Further studies are needed to evaluate ideal thoracic to lumbar vertebral column length ratios in miniature dachshunds.

Activated Chicken Gamma Delta T Cells Are Involved in Protective Immunity against Marek's Disease.

Gamma delta (γδ) T cells play a significant role in the prevention of viral infection and tumor surveillance in mammals. Although the involvement of γδ T cells in Marek's disease virus (MDV) infection has been suggested, their detailed contribution to immunity against MDV or the progression of Marek's disease (MD) remains unknown. In the current study, T cell receptor (TCR)γδ-activated peripheral blood mononuclear cells (PBMCs) were infused into recipient chickens and their effects were examined in the context of tumor formation by MDV and immunity against MDV. We demonstrated that the adoptive transfer of TCRγδ-activated PBMCs reduced virus replication in the lungs and tumor incidence in MDV-challenged chickens. Infusion of TCRγδ-activated PBMCs induced IFN-γ-producing γδ T cells at 10 days post-infection (dpi), and degranulation activity in circulating γδ T cell and CD8α+ γδ T cells at 10 and 21 dpi in MDV-challenged chickens. Additionally, the upregulation of IFN-γ and granzyme A gene expression at 10 dpi was significant in the spleen of the TCRγδ-activated PBMCs-infused and MDV-challenged group compared to the control group. Taken together, our results revealed that TCRγδ stimulation promotes the effector function of chicken γδ T cells, and these effector γδ T cells may be involved in protection against MD.

Synthesis and pharmacological evaluation of newly detected synthetic cannabinoid receptor agonists AB-4CN-BUTICA, MMB-4CN-BUTINACA, MDMB-4F-BUTICA, MDMB-4F-BUTINACA and their analogs.

Synthetic cannabinoid receptor agonists (SCRAs) continue to make up a significant portion new psychoactive substances (NPS) detected and seized worldwide. Due to their often potent activation of central cannabinoid receptors in vivo, use of SCRAs can result in severe intoxication, in addition to other adverse health effects. Recent detections of AB-4CN-BUTICA, MMB-4CN-BUTINACA, MDMB-4F-BUTICA and MDMB-4F-BUTINACA mark a continuation in the appearance of SCRAs bearing novel tail substituents. The proactive characterization campaign described here has facilitated the detection of several new SCRAs in toxicological case work. Here we detail the synthesis, characterization, and pharmacological evaluation of recently detected SCRAs, as well as a systematic library of 32 compounds bearing head, tail, and core group combinations likely to appear in future. In vitro radioligand binding assays revealed most compounds showed moderate to high affinity at both CB1 (pK i = < 5 to 8.89 ± 0.09 M) and CB2 (pK i = 5.49 ± 0.03 to 9.92 ± 0.09 M) receptors. In vitro functional evaluation using a fluorescence-based membrane potential assay showed that most compounds were sub-micromolar to sub-nanomolar agonists at CB1 (pEC50 = < 5 to 9.48 ± 0.14 M) and CB2 (pEC50 = 5.92 ± 0.16 to 8.64 ± 0.15 M) receptors. An in silico receptor-ligand docking approach was utilized to rationalize binding trends for CB2 with respect to the tail substituent, and indicated that rigidity in this region (i.e., 4-cyanobutyl) was detrimental to affinity.

Projecting biodiversity benefits of conservation behavior-change programs.

Biodiversity loss is driven by human behavior, but there is uncertainty about the effectiveness of behavior-change programs in delivering benefits to biodiversity. To demonstrate their value, the biodiversity benefits and cost-effectiveness of behavior changes that directly or indirectly affect biodiversity need to be quantified. We adapted a structured decision-making prioritization tool to determine the potential biodiversity benefits of behavior changes. As a case study, we examined two hypothetical behavior-change programs--wildlife gardening and cat containment--by asking experts to consider the behaviors associated with these programs that directly and indirectly affect biodiversity. We assessed benefits to southern brown bandicoot (Isoodon obesulus) and superb fairy-wren (Malurus cyaneus) by eliciting from experts estimates of the probability of each species persisting in the landscape given a range of behavior-change scenarios in which uptake of the behaviors varied. We then compared these estimates to a business-as-usual scenario to determine the relative biodiversity benefit and cost-effectiveness of each scenario. Experts projected that the behavior-change programs would benefit biodiversity and that benefits would rise with increasing uptake of the target behaviors. Biodiversity benefits were also predicted to accrue through indirect behaviors, although experts disagreed about the magnitude of additional benefit provided. Scenarios that combined the two behavior-change programs were estimated to provide the greatest benefits to species and be most cost-effective. Our method could be used in other contexts and potentially at different scales and advances the use of prioritization tools to guide conservation behavior-change programs.

Proyección de los beneficios para la biodiversidad obtenidos de los programas de cambios de comportamiento de conservación Resumen La pérdida de la diversidad biológica es causada por el comportamiento humano, pero existe incertidumbre sobre la efectividad que tienen los programas de cambio de comportamiento para otorgar beneficios a la biodiversidad. Para demostrar el valor que poseen, los beneficios para la biodiversidad y la rentabilidad de los cambios de comportamiento que afectan directa o indirectamente a la biodiversidad necesitan ser cuantificados. Adaptamos una herramienta de priorización de toma de decisiones estructurada para determinar el potencial de los beneficios para la biodiversidad obtenidos de los cambios de comportamiento. Como estudio de caso, examinamos dos programas hipotéticos de cambio de comportamiento, la jardinería silvestre y la contención de gatos, mediante la petición a expertos de considerar los comportamientos asociados con estos programas que directa o indirectamente afectan a la biodiversidad. Evaluamos los beneficios para el bandicut café (Isoodon obeselus) y el reyezuelo supremo (Malurus cyaeneus) mediante la obtención de estimaciones de expertos de la probabilidad de que cada especie persista en el paisaje con una gama establecida de escenarios de cambios de comportamiento en los cuales la aceptación de los comportamientos varió. Después comparamos estas estimaciones con un escenario de situación normal para determinar el beneficio relativo para la biodiversidad y la rentabilidad de cada escenario. Los expertos proyectaron que los programas de cambio de comportamiento beneficiarían a la biodiversidad y que los beneficios aumentarían con la creciente aceptación de los comportamientos deseados. También se pronosticó que los beneficios se acumularían mediante comportamientos indirectos, aunque los expertos estuvieron en desacuerdo sobre la magnitud del beneficio adicional proporcionado. Se estimó que los escenarios que combinaron los dos programas de cambio de comportamiento proporcionarían el mayor beneficio para las especies y serían los más rentables. Nuestro método podría usarse en otros contextos y potencialmente a diferentes escalas y fomenta el uso de herramientas de priorización para orientar a los programas de cambios en el comportamiento de conservación.

Cannabichromene and Δ9-Tetrahydrocannabinolic Acid Identified as Lactate Dehydrogenase-A Inhibitors by in Silico and in Vitro Screening.

Cannabis sativa contains >120 phytocannabinoids, but our understanding of these compounds is limited. Determining the molecular modes of action of the phytocannabinoids may assist in their therapeutic development. Ligand-based virtual screening was used to suggest novel protein targets for phytocannabinoids. The similarity ensemble approach, a virtual screening tool, was applied to target identification for the phytocannabinoids as a class and predicted a possible interaction with the lactate dehydrogenase (LDH) family of enzymes. In order to evaluate this in silico prediction, a panel of 18 phytocannabinoids was screened against two LDH isozymes (LDHA and LDHB) in vitro. Cannabichromene (CBC) and Δ9-tetrahydrocannabinolic acid (Δ9-THCA) inhibited LDHA via a noncompetitive mode of inhibition with respect to pyruvate, with Ki values of 8.5 and 6.5 µM, respectively. In silico modeling was then used to predict the binding site for CBC and Δ9-THCA. Both were proposed to bind within the nicotinamide pocket, overlapping the binding site of the cofactor NADH, which is consistent with the noncompetitive modes of inhibition. Stemming from our in silico screen, CBC and Δ9-THCA were identified as inhibitors of LDHA, a novel molecular target that may contribute to their therapeutic effects.

Terpenoids Commonly Found in Cannabis sativa Do Not Modulate the Actions of Phytocannabinoids or Endocannabinoids on TRPA1 and TRPV1 Channels.

Introduction: Cannabis sativa produces hundreds of bioactive compounds, including cannabinoids and terpenoids. It has been proposed that cannabinoids act in synergy with terpenoids to produce the entourage effect, a concept used to explain the therapeutic benefits of medicinal cannabis. One molecular explanation for the entourage effect is that the terpenoids augment the actions of cannabinoids at their molecular drug targets in cells. We recently reported that terpenoids commonly found in cannabis do not influence the functional effects of Δ9-tetrahydrocannabinol (Δ9-THC) on cannabinoid 1 and cannabinoid 2 receptors. The present study aimed to extend on this research by examining whether terpenoids influence the effects of phytocannabinoids and endocannabinoids on human transient receptor potential ankyrin 1 (hTRPA1) and human transient receptor potential vanilloid 1 (hTRPV1) channels heterologously expressed in mammalian cells. Materials and Methods: The activity of terpenoids, phytocannabinoids, and endocannabinoids was assessed in inducible HEK Flp-In T-Rex cells transfected with hTRPA1 and hTRPV1 channels, respectively. Real-time changes in intracellular calcium ([Ca]i) were measured using the Calcium 5 dye and a FlexStation 3 plate reader. Results: α-pinene, ß-pinene, ß-caryophyllene, linalool, limonene, ß-myrcene or α-humulene did not affect [Ca]i in hTRPA1 and hTRPV1 overexpressing cells. Cinnamaldehyde (CA), Δ9-THC, and 2-arachidonoylglycerol (2-AG) activated TRPA1 receptors with high efficacy and similar potency (EC50s of ∼10 µM). Capsaicin and anandamide (AEA) activated TRPV1 receptors with an EC50 of 61 nM and 4.3 µM, respectively, but TRPV1 showed no response to Δ9-THC, cannabidiol, and other minor cannabinoids. Terpenoids did not significantly affect the responses of TRPA1 and TRPV1 receptors to submaximal and maximal concentrations of CA and Δ9-THC or the endocannabinoids AEA and 2-AG. Discussion: We could not find any evidence that the terpenoids tested here activate TRPA1 and TRPV1 channels or modulate their activation by Δ9-THC and other agonists, including endocannabinoids.

Mouth rinsing with a sweet solution increases energy expenditure and decreases appetite during 60 min of self-regulated walking exercise.

Carbohydrate mouth rinsing can improve endurance exercise performance and is most ergogenic when exercise is completed in the fasted state. This strategy may also be beneficial to increase exercise capacity and the energy deficit achieved during moderate-intensity exercise relevant to weight control when performed after an overnight fast. Eighteen healthy men (mean (SD); age, 23 (4) years; body mass index, 23.1 (2.4) kg·m-2) completed a familiarisation trial and 3 experimental trials. After an overnight fast, participants performed 60 min of treadmill walking at a speed that equated to a rating of perceived exertion of 13 ("fairly hard"). Participants manually adjusted the treadmill speed to maintain this exertion. Mouth rinses for the experimental trials contained either a 6.4% maltodextrin solution with sweetener (CHO), a taste-matched placebo (PLA), or water (WAT). Appetite ratings were collected using visual analogue scales and exercise energy expenditure and substrate oxidation were calculated from online gas analysis. Increased walking distance during CHO and PLA induced greater energy expenditure compared with WAT (mean difference (90% confidence interval); 79 (60) kJ, P = 0.035, d = 0.24; and 90 (63) kJ, P = 0.024, d = 0.27, respectively). Appetite area under the curve was lower in CHO and PLA than WAT (8 (6) mm, P = 0.042, d = 0.43; and 6 (8) mm, P = 0.201, d = 0.32, respectively). Carbohydrate oxidation was higher in CHO than PLA and WAT (7.3 (6.7) g, P = 0.078, d = 0.47; and 10.1 (6.5) g, P = 0.015, d = 0.81, respectively). This study provides novel evidence that mouth rinsing with a sweetened solution may promote a greater energy deficit during moderate-exertion walking exercise by increasing energy expenditure and decreasing appetite. A placebo effect may have contributed to these benefits.