Mode Of Action Research Articles

Assessment of the genotoxicity of tert-butyl alcohol in an in vivo thyroid comet assay.

Chronic exposure to high (20,000 ppm) concentrations of tert-butyl alcohol (TBA) in drinking water, equivalent to ~2100 mg/kg bodyweight per day, is associated with slight increases in the incidence of thyroid follicular cell adenomas and carcinomas in mice, with no other indications of carcinogenicity. In a recent toxicological review of TBA, the U.S. EPA determined that the genotoxic potential of TBA was inconclusive, largely based on non-standard studies such as in vitro comet assays. As such, the potential role of genotoxicity in the mode of action of thyroid tumors and therefore human relevance was considered uncertain. To address the potential role of genotoxicity in TBA-associated thyroid tumor formation, CD-1 mice were exposed up to a maximum tolerated dose of 1500 mg/kg-day via oral gavage for two consecutive days and DNA damage was assessed with the comet assay in the thyroid. Blood TBA levels were analyzed by headspace GC-MS to confirm systemic tissue exposure. At study termination, no significant increases (DNA breakage) or decreases (DNA crosslinks) in %DNA tail were observed in TBA exposed mice. In contrast, oral gavage of the positive control ethyl methanesulfonate significantly increased %DNA tail in the thyroid. These findings are consistent with most genotoxicity studies on TBA and provide mechanistic support for non-linear, threshold toxicity criteria for TBA. While the mode of action for the thyroid tumors remains unclear, linear low dose extrapolation methods for TBA appear more a matter of policy than science.

A new perspective on therapies involving B-cell depletion in autoimmune diseases.

It has been rediscovered in the last fifteen years that B-cells play an active role in autoimmune etiology rather than just being spectators. The clinical success of B-cell depletion therapies (BCDTs) has contributed to this. BCDTs, including those that target CD20, CD19, and BAFF, were first developed to eradicate malignant B-cells. These days, they treat autoimmune conditions like multiple sclerosis and systemic lupus erythematosus. Particular surprises have resulted from the use of BCDTs in autoimmune diseases. For example, even in cases where BCDT is used to treat the condition, its effects on antibody-secreting plasma cells and antibody levels are restricted, even though these cells are regarded to play a detrimental pathogenic role in autoimmune diseases. In this Review, we provide an update on our knowledge of the biology of B-cells, examine the outcomes of clinical studies employing BCDT for autoimmune reasons, talk about potential explanations for the drug's mode of action, and make predictions about future approaches to targeting B-cells other than depletion.

New Chalcogen-Functionalized Naphthoquinones: Design, Synthesis, and Evaluation, In Vitro and In Silico, against Squamous Cell Carcinoma.

Due to the growth in the number of patients and the complexity involved in anticancer therapies, new therapeutic approaches are urgent and necessary. In this context, compounds containing the selenium atom can be employed in developing new medicines due to their potential therapeutic efficacy and unique modes of action. Furthermore, tellurium, a previously unknown element, has emerged as a promising possibility in chalcogen-containing compounds. In this study, 13 target compounds (9a-i, 10a-c, and 11) were effectively synthesized as potential anticancer agents, employing a CuI-catalyzed Csp-chalcogen bond formation procedure. The developed methodology yielded excellent results, ranging from 30 to 85%, and the compounds were carefully characterized. Eight of these compounds showed promise as potential therapeutic drugs due to their high yields and remarkable selectivity against SCC-9 cells (squamous cell carcinoma). Compound 10a, in particular, demonstrated exceptional selectivity, making it an excellent choice for cancer cell targeting while sparing healthy cells. Furthermore, complementing in silico and molecular docking studies shed light on their physical features and putative modes of action. This research highlights the potential of these compounds in anticancer treatments and lays the way for future drug development efforts.

Mycobactin analogue interacting with siderophore efflux-pump protein: insights from molecular dynamics simulations and whole-cell assays

IntroductionIn response to continued public health emergency of antimicrobial resistance (AMR), a significant key strategy is the discovery of novel mycobacterial efflux-pump inhibitors (EPIs) as potential adjuvants in combination drug therapy. Interest in identifying new chemotypes which could potentially synergize with the existing antibiotics and can be deployed as part of a combination therapy. This strategy could delay the emergence of resistance to existing antibiotics and increase their efficacy against resistant strains of mycobacterial species. In recent decades, notable approaches have been accounted for EPI development and have resulted in the discovery of several EPIs including SQ109 and AU1235. In context, to accelerate newer EPIs with novel mode of action here we have discussed mycobactin analogues and highlighted in silico binding orientation with siderophore efflux-pump proteins MmpL4/5.Methods3-(2-hydroxyphenyl)-5-(aryl)-pyrazoline series was investigated for whole-cell efflux-pump inhibitory activity against Mycobacterium smegmatis and Mycobacterium abscessus. Machine learning and molecular dynamics were performed to construct a MmpL4/5 complex embedded in a lipid bilayer to identify the putative binding site and to predict ligand-protein binding energetics. Furthermore, the identified HIT compound was investigated in synergistic assay with bedaquiline.ResultsCompound Il, 2-(5-(4-fluorophenyl)-4,5-dihydro-1H-pyrazol-3-yl)phenol, was identified as the most potent efflux pump inhibitor against M. smegmatis in whole-cell efflux-pump investigation. Followed HIT Il employed against M. abscessus for efflux-pump inhibition investigations and notable whole-cell efflux-pump inhibitory profile has been observed. The theoretical investigations predicted compound Il to be selective towards MmpL4, with significant hydrogen bonding and π-π stacking interactions effectively blocking a critical Asp-Tyr dyad interaction network necessary for proton translocation. Compound Il with bedaquiline highlighted an additive profile against the M. abscessus pathogen.ConclusionsMD simulations and whole-cell assays are indicating potential development of compound Il as an adjunct to the existing therapeutic regimen against mycobacterial infections.

Endothall and 2,4-D Activity in Milfoil Hybrid (Myriophyllum spicatum × M. sibiricum) when Applied Alone and in Combination

Abstract Eurasian watermilfoil (Myriophyllum spicatum L.) is an invasive aquatic plant that can hybridize with the native Northern watermilfoil (Myriophyllum sibiricum Kom.). These milfoil hybrids (M. spicatum × M. sibiricum) are becoming more prevalent in many lakes where the invasive and the native milfoil co-occur. Hybrid plants are more vigorous than either parent with a faster growth rate and lower sensitivity to some herbicides. The aquatic herbicides, endothall and 2,4-D, provide two effective modes of action (MOA) for management of the hybrids. For more than a decade, these two herbicides have been used in combination as an effective control option and a resistance management strategy. How this combination impacts herbicide movement and efficacy is unknown. Therefore, the objective of this research was to determine the activity of endothall and 2,4-D combined compared to activity applied alone. Absorption and translocation of endothall, 2,4-D and the combination was determined in hybrid plants over a 96-h time course. Endothall bioaccumulation was not impacted when these herbicides were applied in combination; however, 2,4-D accumulation increased by 80%, relative to when 2,4-D was applied alone. Endothall translocation from shoots to roots decreased by almost 50% when applied in combination with 2,4-D (alone = 16.7% ± 2.6; combination = 9.2% ± 1.2). Shoot-to-root translocation of 2,4-D also decreased when the two herbicides were applied in combination (24.8% ± 2.6 when applied alone to only 3.93% ± 0.4 when in the presence of endothall). This research demonstrates that combining herbicides can significantly impact herbicide activity in plants. Future research is needed to determine if this reduced translocation negatively impacts operational effectiveness when these herbicides applied in combination.

Evaluation of in vitro antioxidant activities, total phenolic and elemental contents of common herbs and spices (Moringa oleifera leaves, Allium sativum (Garlic) and Momordica charantia (ejinrin) leaves) in South-West Nigeria

Abstract Natural remedies are being widely utilized in some disease’s conditions including cancer, diabetes, neurodegenerative diseases, atherosclerosis hypertension and other cardiovascular diseases. The therapeutic intervention of medicinal plants and spices cannot be overemphasized in the management and control of diseases but the exact modes of action of these herbs and plants have not been fully elucidated. This research work was designed to study the expression of selected elements, zinc (Zn), iron (Fe), copper (Cu), selenium (Se), lead (Pb) and cadmium (Cd); the phytochemical and the in vitro antioxidant properties of ethanolic extracts of Moringa oleifera leaves, Allium sativum (Garlic) and Momordica charantia (ejinrin) leaves for their potential involvement in the prevention and management of cardiovascular diseases and cancer through different chemical methods. In vitro antioxidant properties were assayed by investigating 2,2-diphenyl-2-picryl-hydrazyl (DPPH) free radicals scavenging potentials and Ferric Reducing Antioxidant Potential (FRAP); total phenolic content was determined by using Folin-Ciocalteu assay and the elemental contents of these extracts was investigated through the use of Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES). All tests were run in triplicates and analysed. Our results revealed that the ethanolic extracts of M. oleifera leaves, A. sativum (Garlic) and M. charantia (ejinrin) leaves possess significant antioxidant activities and express important and beneficial elements (in marginal and trace amounts) and phytochemicals. These results suggest that the medicinal attributes of these plants may be linked to the radical scavenging abilities, beneficial elements and phytochemicals expression of their ethanolic extracts which may likely be a good direction in the area of drug discovery and development.

How Much Potential Do Nucleoside Analogs Offer to Combat Human Corona Viruses?

Nucleoside analogs (NAs) have been extensively examined as plausible antiviral agents in recent years, in particular since the outbreak of the global pandemic of COVID-19 in 2019. In this review, the structures and antiviral properties of over 450 NAs are collected according to the type of virus, namely SARS-CoV, SARS-CoV-2, MERS-CoV, HCoV-OC43, HCoV-229E, and HCoV-NL63. The activity of the NAs against HCoV-related enzymes is also presented. Selected studies dealing with the mode of action of the NAs are discussed in detail. The repurposing of known NAs appears to be the most extensively investigated scientific approach towards efficacious anti-HCoV agents. The recently reported de novo-designed NAs seem to open up additional approaches to new drug candidates.

Design, Synthesis, Antifungal Activity, and Action Mechanism of Pyrazole-4-carboxamide Derivatives Containing Oxime Ether Active Fragment As Succinate Dehydrogenase Inhibitors.

The dearomatization at the hydrophobic tail of the boscalid was carried out to construct a series of novel pyrazole-4-carboxamide derivatives containing an oxime ether fragment. By using fungicide-likeness analyses and virtual screening, 24 target compounds with theoretical strong inhibitory effects against fungal succinate dehydrogenase (SDH) were designed and synthesized. Antifungal bioassays showed that the target compound E1 could selectively inhibit the in vitro growth of R. solani, with the EC50 value of 1.1 μg/mL that was superior to that of the agricultural fungicide boscalid (2.2 μg/mL). The observations by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) demonstrated that E1 could reduce mycelial density and significantly increase the mitochondrial number in mycelia cytoplasm, which was similar to the phenomenon treated with boscalid. Enzyme activity assay showed that the E1 had the significant inhibitory effect against the SDH from R. solani, with the IC50 value of 3.3 μM that was superior to that of boscalid (7.9 μM). The mode of action of the target compound E1 with SDH was further analyzed by molecular docking and molecular dynamics simulation studies. Among them, the number of hydrogen bonds was significantly more in the SDH-E1 complex than that in the SDH-boscalid complex. This research on the dearomatization strategy of the benzene ring for constructing pyrazole-4-carboxamides containing an oxime ether fragment provides a unique thought to design new antifungal drugs targeting SDH.

Thermal proteome profiling reveals insight to antiproliferative and pro-apoptotic effects of Lagunamide A in the modulation of DNA damage repair.

Lagunamide A is a biologically active natural product with a yet unidentified molecular mode of action. Cellular studies revealed that lagunamide A is a potent inhibitor of cancer cell proliferation, promotes apoptosis and causes mitochondrial dysfunction. To decipher the cellular mechanism responsible for these effects, we utilized thermal protein profiling (TPP) and identified EYA3 as a stabilized protein in cells upon lagunamide A treatment. EYA3, involved in the DNA damage repair process, was functionally investigated via siRNA based knockdown studies and corresponding effects of lagunamide A on DNA repair were confirmed. Furthermore, we showed that lagunamide A sensitized tumor cells to treatment with the drug doxorubicin highlighting a putative therapeutic strategy.

Screening and In Silico Analyses of the Yeast Saccharomyces cerevisiae Σ1278b Bank Mutants Using Citral as a Natural Antimicrobial.

The antimicrobial function of citral, one of the main compounds of the essential oils (EO) of the Citrus genus, and widely used by the food industry toward spoilage yeast, was previously proven. In this study, the possible mode of action of citral against yeast cells was evaluated by using a global deletome approach. Firstly, the suitability of Saccharomyces cerevisiae Σ1278b to serve as model yeast was assessed by determining its sensitivity to citral (MIC = 0.5 μL/mL). Subsequently, the complete library of Σ1278b haploid mutants deleted in 4019 non-essential genes was screened to identify potential molecular targets of citral. Finally, the deleted genes in the 590 mutants showing increased citral resistance was analyzed with an in-silico approach (Gene Ontology). The significantly enriched GO Terms were "cytoplasm", "vacuole", and "mitochondrion" (cellular components); "catalytic activity" (molecular function); "pseudohyphal growth" (biological process). For molecular function, resistant mutants were grouped into thiosulfate sulfur transferase activity, transferase activity, and oxidoreductase activity; for cellular components, resistant mutants were grouped as: cytoplasm, intracellular organelle, membrane-bounded organelle, mitochondrion, organelle membrane, and vacuole; and finally, with regard to biological process, deleted genes were grouped as: pseudohyphal growth, mitochondrion organization, lipid metabolic process, DNA recombination and repair, and proteolysis. Interestingly, many identified genes were associated with the cellular response to oxidative stress and ROS scavenging. These findings have important implications for the development of citral-based antimicrobials and the elucidation of its mechanism of action.

Regulatory Framework for Drug-Device Combination Products in the United States, Europe, and Korea.

Combination products (CPs) combine two or more product types such as drugs, devices, and/or biological products for increased safety and clinical effectiveness. The emergence of innovative CPs poses new challenges for regulatory agencies in assigning jurisdiction for premarket review and oversight. In US, the 1990 Safe Medical Devices Act defines and provides classification criteria for CPs, and the US government has developed a regulatory process through multiple acts, including the 21st Century Cures Act of 2016. Meanwhile, regulators in the European Union (EU) and the Republic of Korea have recently recognized the importance of premarket pathways for CPs. The European Medicines Agency (EMA) has issued guidelines and explanations on compliance issues related to drug-device CPs under MDR. EMA doesn't have the definitions of CPs, but uses the term drug-device combination products (drug-device CPs). CPs are categorized as drugs or medical devices, which follow their relevant regulatory framework. The Ministry of Food and Drug Safety (MFDS) in Korea has legal definitions of CPs under the Notice of the MFDS. CPs are designated as drugs or medical devices according to their primary mode of actions (PMOA) and follow regulatory processes through the framework of drugs or medical devices. This study aims to comprehensively summarize the regulatory oversight of CPs by analyzing the regulatory policies and legal frameworks in the US, the EU, and Korea. The regulatory challenges encountered when developing CPs are also discussed. With specific emphasis on the combination of drugs and devices, this study provides in-depth insights into the regulatory landscape, shedding light on the unique challenges associated with the development of CPs for this particular intersection of drugs and devices.

Suppression of Hop Downy Mildew as Influenced by the Timing of Selected Fungicides

Downy mildew of hop is a disease that is controlled by the implementation of cultural practices along with the application of fungicides, as there are few cultivars with high levels of resistance to this disease. We conducted studies in western Oregon to elucidate the effect of using synthetic fungicides of various modes of action early versus late in disease management programs when rotated with a copper-based fungicide. When analyzed over the 4 years of this study, there were independent effects from both the specific fungicide mode of action and application timing on disease control. The use of cymoxanil + copper hydroxide or mandipropamid in a program reduced the area under the disease progress curve by 66.3 or 59.7 units (17.7 or 19.6%), respectively, as compared with phosphorous acid. Using synthetic fungicides in the first two applications in a program provided better disease control than using the same fungicides in the third and fourth applications, reducing the area under the disease progress curve by 47.8 units (14.4%). By simply applying modes of action found to be more efficacious earlier in the growing season, the severity of downy mildew was reduced as much as 32% without increasing the total number of applications made.

Buparvaquone Induces Ultrastructural and Physiological Alterations Leading to Mitochondrial Dysfunction and Caspase-Independent Apoptotic Cell Death in Leishmania donovani.

Leishmaniasis is a neglected tropical disease (endemic in 99 countries) caused by parasitic protozoa of the genus Leishmania. As treatment options are limited, there is an unmet need for new drugs. The hydroxynaphthoquinone class of compounds demonstrates broad-spectrum activity against protozoan parasites. Buparvaquone (BPQ), a member of this class, is the only drug licensed for the treatment of theileriosis. BPQ has shown promising antileishmanial activity but its mode of action is largely unknown. The aim of this study was to evaluate the ultrastructural and physiological effects of BPQ for elucidating the mechanisms underlying the in vitro antiproliferative activity in Leishmania donovani. Transmission and scanning electron microscopy analyses of BPQ-treated parasites revealed ultrastructural effects characteristic of apoptosis-like cell death, which include alterations in the nucleus, mitochondrion, kinetoplast, flagella, and the flagellar pocket. Using flow cytometry, laser scanning confocal microscopy, and fluorometry, we found that BPQ induced caspase-independent apoptosis-like cell death by losing plasma membrane phospholipid asymmetry and cell cycle arrest at sub-G0/G1 phase. Depolarization of the mitochondrial membrane leads to the generation of oxidative stress and impaired ATP synthesis followed by disruption of intracellular calcium homeostasis. Collectively, these findings provide valuable mechanistic insights and demonstrate BPQ's potential for development as an antileishmanial agent.

Centipeda minima (L.) A. Braun & Asch. and its representative active compound alleviate DSS-induced ulcerative colitis via inhibition of NLRP3 inflammasome activation and regulation of gut microbiota

Ulcerative colitis (UC) is a prevalent chronic inflammatory bowel disease (IBD), which is related to the occurrence of inflammation and intestinal flora disorder. Centipeda minima (L.) A. Braun & Asch. (CM for short hereafter) is commonly used to treat IBD in folk medicine. However, the anti-UC mechanisms of CM and its representative active compound(s) are not yet fully understood. Here, we sought to investigate the mode of action and the underlying mechanism of CM as an anti-UC agent, as well as to identify its potential bioactive compound(s). Results showed that 50% ethanol extract of CM (CME) significantly improved the pathological morphology and reduced the protein levels of NLRP3 both in DSS-induced UC mice model and lipopolysaccharide (LPS)-stimulated RAW264.7 cell model. Additionally, it decreased the release of proinflammatory cytokines including TNF-α, IL-18 and IL-1β. These results suggest that the anti-UC effect of CM is at least partially attributed to the inhibition of NLRP3 inflammasome activation. Furthermore, luteolin, a representative active compound in CM, exhibited significant improvement of the pathological changes, reduced the release the of proinflammatory cytokines, downregulated NLRP3 inflammasome activation, enhanced intestinal barrier, and modulated gut microbiota in vivo. This study provides a molecular rationale for the traditional use of CM in treating UC, and offers a pharmacological basis for the development of modern anti-UC agents derived from CM.

99 Phytochemicals as medical growth promoter replacement on growth performance in wean to finisher pigs

Abstract Stress can disturb the ecological balance of the microbiota residing in the GI tract of piglets leading to dysbiosis. This unhealthy imbalance causes the piglets to be more vulnerable to deleterious effects induced by opportunistic pathogens. Antibiotics and increased zinc levels have often been used as dietary supplements to help minimize these stressors in pigs. Emerged evidence suggests that phytochemicals can promote growth by boosting nutrient digestibility, immune cell activity, and possessing bactericidal characteristics. We evaluated two phytochemicals formulations as safer and more environmentally friendly dietary alternatives for postweaning pigs. We used a total of 240 weaned piglets [average body weight (BW): 6.26 ± 0.36 kg] which were blocked (n = 8) by BW and then randomly assigned to 1 of 5 dietary treatments. Treatments were negative control (NC), Mecadox 10 (Phibro Animal Health Corp., Teaneck, NJ) + ZnO positive control (PC), Mecadox + phytochemical 1 (Lippia origanoides; Phyt1+AB), phytochemical 1 alone (Phyt1), and phytochemical 2 (Lippia origanoides + Rosmarinus officinalis + Sodium humates; Phyt2). The pigs were fed a 7-phase feeding regime: Nursery phase (NP) 1, 2, and 3, Grower phase 1 and 2 (Grower), and Finisher phase 1 and 2 (Finisher). The antibiotic and ZnO were withdrawn at the end of NP3, while Phyt1 and Phyt2 remained in feed throughout the entire study period. Data were analyzed using Mixed procedure of SAS (Cary, NC) with treatments as the main effect and BW blocks as the random effect. Pen was the experimental unit. Supplemental phytochemicals (Phyt1+AB, Phyt1, Phyt2) had greater average daily gain (ADG) and gain to feed ratio (G:F) during NP1 when compared with the pigs fed NC, but were still less than pigs fed PC (P < 0.0001). Pigs fed PC maintained a greater ADG during nursery overall than NC, Phyt1, and Phyt2 with pigs fed Phyt1+AB as intermediate (P < 0.0001). Phyt1 exerted an adverse effect (Phyt1+AB and Phyt1) on ADG during relatively mild stress in Grower phase (Grower, P = 0.0146), and resulted in the lighter BW at the end of Grower than pigs fed PC (P = 0.031). Notably, the pigs experienced summer heat stress during the Finisher phase, of which, the presence of phytochemicals minimized. Pigs fed Phyt2 had a greater ADG during Finisher than pigs fed NC with pigs fed PC, Phyt1+AB, and Phyt1 as intermediates (Finisher, P = 0.0419), while pigs fed Phyt1+AB and Phyt2 had greater G:F when compared with pigs fed PC and NC with pigs fed Phyt1 as intermediate (Finisher, P = 0.0032). Collectively, we demonstrated that phytochemical 2 can be a safer medication diet replacement to help pigs sustain weaning and heat stress. The mode of action and the optimum concentrations of these phytochemicals to achieve maximum benefits need further investigation.

233 Inclusion of a multispecies fungal feed additive in forage-based diets fed to beef cattle: Effects on growth performance and ruminal fermentation

Abstract The objective was to evaluate the effect of adding a multispecies fungal feed additive (MFA; Aspergillus flavus, Aspergillus niger, Aspergillus oryzae) to forage-based diets fed to beef cattle on growth performance and ruminal fermentation. Experiment 1 evaluated the effect of supplementing Angus × SimAngus-crossbred steers [n = 80; initial body weight (BW) = 370 ± 44 kg] with varying doses of an MFA in a randomized complete block design. Steers were blocked by BW and housed in 8 feedlot pens (randomly assigned 2 pens per treatment) where a hay-based diet was fed for 122 d. The hay-based diet was offered with the inclusion of the MFA at 0.02, 0.04, and 0.08 % of the diet dry matter (DM), or without the inclusion of the MFA (CON). Steers were weighed at 28-d intervals. For experiment 2, ruminally cannulated Angus × SimAngus cows (n = 4; initial BW = 569 ± 21 kg) were used in a crossover design with two 21-d study periods to evaluate the effect of dietary inclusion of an MFA on in situ digestion and rumen metabolites. Treatments consisted of a forage-based diet with (WFA) or without (CON) the inclusion of the MFA. In situ digestion was evaluated on d 5, 10, and 20 throughout each period. In-situ bags were incubated in the rumen for 0 to 36 h, sampling every 4 h. On d 20, samples of ruminal fluid were collected to analyze short-chain fatty acids (SCFA) concentration. All data were analyzed using the PROC MIXED procedure of SAS. For experiment 1, a polynomial contrast was used. For experiment 2, the interaction between period and treatment was not significant (P > 0.45). In Experiment 1, the final BW, average daily gain (ADG), and gain to feed ratio (G:F) changed cubically (P ≤ 0.04; Table 1) as dietary inclusion of the MFA increased. Steers supplemented with the MFA at a rate of 0.02% of the diet DM were heavier at the end of the 122-d study period and had the greatest ADG and G:F among treatments. No differences between treatments (P ≥ 0.37) were observed for dry matter intake. In Experiment 2, no treatment × time interactions and treatment effects were detected (P ≥ 0.17; Table 2) for the in-situ disappearance rate and degradation kinetics of the diet DM, neutral detergent fiber, and acid detergent fiber. In addition, MFA supplementation did not affect (P ≥ 0.13; Table 3) the SCFA profile and the total SCFA content in the rumen fluid. Results from these studies indicate that dietary inclusion of an MFA might be an effective alternative to increase the feed efficiency of beef cattle-fed forage-based diets. However, the precise mode of action warrants further investigations.

126 A nutritional solution based on calcidiol (25-OH-D3, HyD) and triterpenoid affects muscle protein synthesis pathways in grower-finisher pigs

Abstract To boost profitability in pig production, it is important to achieve increased lean growth rates and maximize efficiency, especially in grower-finisher pigs where feed intakes are high. Molecular research, especially the study of gene expression, is a proven tool for understanding the precise mechanisms behind muscle tissue growth, including a recent focus on pathways such as the mammalian target of rapamycin (mTOR). In the current work, gene expression analyses were used to study the mode of action of the combination of calcidiol (25-OH-D3, HyD) and triterpenoid, which has shown significant growth and efficiency improvements in grower-finisher pigs (McCormack et al., 2024). Three experimental diets were evaluated for 7 wk; Positive control: 0.7% total Ca, 0.36% total P, vit D3 at 1500IU/kg feed (PC), negative control: PC diet + 30% reduced Ca and 10% reduced P (NC), and NC diet with 1,500IU D3 in calcidiol form (25-OH-D3, HyD) and triterpenoid (NC+). At 19 wk of age, 16 animals per treatment were sacrificed and tissue samples were taken from the longissimus muscle and preserved in RNA later solution. The RNA was extracted, quality control checked, and mRNA sequenced using the Illumina platform with 150 base pair paired-end sequencing and 30M read depth. The sequenced data were aligned to the Sus scrofa reference genome (Sscrofa11.1) using Kallisto. Data were normalized and analyzed for differentially expressed genes (DEGs) between treatments using DeSeq2. Genes were assigned to biological pathways using Biofractal’s customisation of the Reactome pathway database. For statistical analysis, RNA integrity number and treatment were included in the model. In the PC compared with the NC, there were 71 DEGs using a false discovery rate (FDR) of < 0.10. Altered pathways included the inhibition of cellular stress pathways and increased cell turnover in the muscle of PC compared with NC pigs. In the NC+ compared with the NC, there were 155 DEGs (FDR < 0.10). The most significantly upregulated gene in the NC+ treatment was adenylate cyclase (FDR < 0.001), an upstream regulator of several pathways related to energy metabolism and muscle protein synthesis. This mechanism was confirmed in the pathway analysis, which found that the NC+ treatment activated PKA-mediated phosphorylation of CREB, a pathway that inhibits muscle atrophy. Furthermore, the NC+ treatment activated several pathways related to muscle growth, including the mammalian target of rapamycin complex 1 (mTORC1). In this study the addition of calcidiol (25-OH-D3, HyD) and triterpenoid to the diet was found to specifically affect molecular pathways in the muscle of grower-finisher pigs involved in both the inhibition of muscle breakdown as well as activating muscle growth.

Antifungal Activity, Mode of Action, and Cytotoxicity of 4-chlorobenzyl p-coumarate: a Promising New Molecule.

Fungal infections represent a serious health problem worldwide. The study evaluated the antifungal activity of 4-chlorobenzyl p-coumarate, an unprecedented semi-synthetic molecule. Docking molecular and assay experiments were conducted to determine the Minimum Inhibitory Concentration (MIC) and Minimum Fungicidal Concentration (MFC), mode of action, effect on growth, fungal death kinetics, drug association, effects on biofilm, micromorphology, and against human keratinocytes. The investigation included 16 strains ofCandidaspp, includingC. albicans,C. krusei,C. glabrata,C. tropicalis,C. dubliniensis,C. lusitaniae,C. utilis,C. rugosa,C. guilhermondi, andC. parapsilosis. Docking analysis predicted affinity between the molecule and all tested targets. MIC and MFC values ranged from 3.9 µg/mL (13.54 µM) to 62.5 µg/mL (217.01 µM), indicating a probable effect on the plasma membrane. The molecule inhibited growth from the first hour of testing. Association with nystatin proved to be indifferent. All concentrations of the molecule reduced fungal biofilm. The compound altered fungal micromorphology. The tested compound exhibited an IC50 of 7.90 ± 0.40 µg/mL (27.45 ± 1.42 µM) for keratinocytes. 4-chlorobenzyl p-coumarate showed strong fungicidal effects, likely through its action on the plasma membrane and alteration of fungal micromorphology, and mildly cytotoxic to human keratinocytes.

"Thank You for Sharing this Fantastic Performance"

YouTube is one of the most important music media in the world today. Musical practice is reflected there in a variety of forms, from sound recordings to music videos to tutorials. The media environment that YouTube creates as a platform inscribes itself – to some degree – in the actions of those who use it. This is also the case with the YouTube drummer phenomenon, which this article examines in more detail in terms of the interplay between persona construction, music-making, media presentation, and transmedia content concatenation. The thesis is pursued that the practice of YouTube drummers creates a singular interference space of conventions, modes of action, and social roles, which in some places has novelty value with regard to music-related persona constructions and their implementation in platform-based audiovisual media, and which transcends media boundaries. The specificity of the persona construction essentially results from the mediating position of the drum-playing actors: they are not only performers who, like social media influencers, push into the light of publicity, but at the same time musical experts and instructors, quasi-curatorial music communicators who select, prepare, and present music of the past, and intermediaries of the corporeality of popular music. The analyses presented are based on exploratory investigations of the YouTube drummer field itself and a case study.

Gold Nanoparticle Synthesis and its Mode of Action as an Effective Therapy against Breast Carcinoma: Analysis of Traditional Methods and their Comparison with New Methods

Abstract: Breast cancer is one of the most widespread and lethal malignancies afflicting females globally. The global incidence of breast cancer is on the rise, with an anticipated 4.4 million cases by 2070. Notably, breast cancer constitutes approximately 25% of all cancer diagnoses. In the realm of biomedical sciences, nanotechnology has ushered in a transformative era, particularly in cancer therapy and diagnostics. Nanoparticles offer the capability to tailor specific sizes, with elevated surface-to-volume ratios proving advantageous for drug distribution by ensuring a substantial medicines loading volume. Gold nanoparticles exhibit a remarkable selectivity for cancer cells, primarily attributed to the heightened permeability and retaining effects. This study aims to assess the effectiveness of gold nanoparticles in breast cancer therapy, comparing them with traditional techniques from previous research and elucidating their mechanism of action. Data for analysis were sourced from various platforms, including Web of Science and Google Scholar, comprising previously published information. Analysis revealed that historical studies employed conventional techniques for breast cancer therapy. However, contemporary approaches now favor nanomedicine, incorporating diverse drugs and delivery systems, surpassing traditional methods. In summary, nanomedicine emerges as an exceptionally effective mode of breast cancer treatment when juxtaposed with conventional approaches. Previous research also underscores the efficacy of gold nanoparticles as a viable treatment modality. Currently, a spectrum of nanomedicines, including gold nanoparticles, serves as a promising avenue for suppressing tumor development in breast cancer patients