Potential Mode Of Action Research Articles

Ferrocenyl β-Diketonate Compounds: Extended Ring Systems for Improved Anticancer Activity.

A library of ferrocenyl β-diketonate compounds with varying degrees of aromatic functionality have been synthesized and fully characterized. This includes cyclic voltammetry and the analysis of four new structures by single crystal X-ray diffraction. The compounds cytotoxic potential has been determined by MTT screening against pancreatic carcinoma (MIA PaCa-2), ovarian adenocarcinoma (A2780), breast adenocarcinomas (MDA-MB-231 and MCF-7) and normal epithelial retinal (ARPE-19). The compounds show a general trend, where increasing the number of aromatic rings in the molecule yields an increase in cytotoxicity and follows the trend anthracenyl > naphthyl > phenyl > methyl. The compounds are particularly sensitive to the triple negative cancer cell line MDA-MB-231, and the potential modes of action have been studied by production of reactive oxygen species using fluorescence microscopy and cell morphology using Scanning Electron Microscopy. All assays highlight the ferrocenyl β-diketonate with an anthracenyl substituent to be the lead compound in this library. The decomposition was also observed within cells, to give a cytotoxic fluorescent molecule, which has been visualized by confocal microscopy.

Disruption of tRNA biogenesis enhances proteostatic resilience, improves later-life health, and promotes longevity.

tRNAs are evolutionarily ancient molecular decoders essential for protein translation. In eukaryotes, tRNAs and other short, noncoding RNAs are transcribed by RNA polymerase (Pol) III, an enzyme that promotes ageing in yeast, worms, and flies. Here, we show that a partial reduction in Pol III activity specifically disrupts tRNA levels. This effect is conserved across worms, flies, and mice, where computational models indicate that it impacts mRNA decoding. In all 3 species, reduced Pol III activity increases proteostatic resilience. In worms, it activates the unfolded protein response (UPR) and direct disruption of tRNA metabolism is sufficient to recapitulate this. In flies, decreasing Pol III's transcriptional initiation on tRNA genes by a loss-of-function in the TFIIIC transcription factor robustly extends lifespan, improves proteostatic resilience and recapitulates the broad-spectrum benefits to late-life health seen following partial Pol III inhibition. We provide evidence that a partial reduction in Pol III activity impacts translation, quantitatively or qualitatively, in both worms and flies, indicating a potential mode of action. Our work demonstrates a conserved and previously unappreciated role of tRNAs in animal ageing.

Synthesis, Antimicrobial and Antibiofilm Activities of Silver(I) Complexes with N-Alkylbenzimidazole Derivatives and Their Molecular Docking Study

This study investigates the antimicrobial and antibiofilm potential of three silver(I) complexes derived from N-alkylbenzimidazole derivatives: bis[(Z)-1-styryl-benzimidazole]silver(I) nitrate (4), bis[(E)-1-styryl-benzimidazole]silver(I) nitrate (5), and bis(1-cinnamyl-benzimidazole)silver(I) nitrate (6). Detailed synthesis and characterization of these complexes are followed by assessments of their efficacy against Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aeruginosa, Escherichia coli, and Candida albicans. Among the tested compounds, complex 6 exhibited the highest antimicrobial activity, with a minimal inhibitory concentration of 6 µmol/L against P. aeruginosa, and significant antibiofilm activity, achieving 67.72% inhibition at a concentration of 3 µmol/L. These interesting findings led us to conduct a molecular docking study to understand the mechanisms of action by investigating the interactions between the silver(I) complexes and some key protein targets involved in bacterial and fungal biofilm formation, including (p)ppGpp synthetase/hydrolases, FtsZ proteins, and pyruvate kinases. This comprehensive approach, combining experimental and computational analyses derived from N-alkylbenzimidazole derivatives, particularly complex 6, which exhibits remarkable efficacy against various pathogens, reveals promising therapeutic applications of these silver(I) complexes and advances our understanding of their potential mode of action against biofilm-associated infections.

Oxidative Stress-induced Hormonal Disruption in Male Reproduction.

Research into the impacts of oxidative stress (OS), and hormonal balance on reproductive potential has increased over the last 40 years possibly due to rising male infertility. Decreased antioxidant levels and increased OS in tissues result from hormonal imbalance, which in turn leads to male infertility. Increased reactive oxygen species (ROS) generation in seminal plasma has been linked to many lifestyle factors such as alcohol and tobacco use, toxicant exposure, obesity, varicocele, stress, and aging. This article provides an overview of the crosslink between OS and gonadal hormone disruption, as well as a potential mode of action in male infertility. Disrupting the equilibrium between ROS generation and the antioxidant defense mechanism in the male reproductive system may affect key hormonal regulators of male reproductive activities. Unchecked ROS production may cause direct injury on reproductive tissues or could disrupt normal regulatory mechanisms of the hypothalamic-pituitary-gonadal (HPG) axis and its interaction with other endocrine axes, both of which have negative effects on male reproductive health and can lead to male infertility.

Network pharmacology, molecular docking, and in vitro study on Aspilia pluriseta against prostate cancer

BackgroundCurrent prostate cancer treatments are associated with life-threatening side effects, prompting the search for effective and safer alternatives. Aspilia pluriseta Schweinf. ex Engl. has previously shown anticancer activity in lung and liver cancer cell lines. This study investigated its potential for prostate cancer.MethodsA crude extract of A. pluriseta root was prepared using dichloromethane/methanol (1:1 v/v) and partitioned into hexane, ethyl acetate, and water fractions. The MTT assay was used to assess the antiproliferative activity of the fractions. The active fractions were tested at 6.25–200 µg/ml on human prostate cancer DU-145 cells and non-cancerous Vero E6 cells. Qualitative phytochemical and gas chromatography-mass spectrometry (GC-MS) analyses were conducted to identify chemical compounds. Network pharmacology was employed to predict molecular targets and modes of action of the identified chemical compounds, with subsequent validation through molecular docking and real-time PCR.ResultsActive extracts included crude dichloromethane/methanol, hexane, and ethyl acetate fractions, inhibiting DU-145 cell proliferation with IC50 values of 16.94, 20.06, and 24.14 µg/ml, respectively. Selectivity indices were determined to be 6.04 (crude), 3.62 (hexane), and 6.68 (ethyl acetate). Identified phytochemicals comprised phenols, terpenoids, flavonoids, tannins, sterols, and saponins. GC-MS analysis revealed seventy-nine (79) compounds, with seven (7) meeting ideal drug candidate parameters; their hub gene targets included MAPK3, MAPK1, IL6, TP53, ESR1, PTGS2, MMP9, MDM2, AR, and MAP2K1, implicating regulation of PI3K/Akt, MAPK, and p53 signaling pathways as potential modes of action. Core compounds such as 1-heneicosanol, lanosterol, andrographolide, and retinoic acid exhibited strong binding activities, particularly lanosterol with MAPK21 (-9.7 kcal/mol), ESR1 (-8.9 kcal/mol), and MAPK3 (-8.8 kcal/mol). Treatment with A. pluriseta downregulated AR expression and upregulated p53, while also downregulating CDK1 and BCL-2 and upregulating caspase-3.ConclusionsA. pluriseta extracts inhibited DU-145 cell growth without causing cellular toxicity, suggesting great potential for development as an anti-prostate cancer agent. However, further in vitro and in vivo experiments are recommended.

‘Microscopic engineering vehicles’ for plants under stress combination

The combination of stresses induced by climate change poses significant risks to crop production. We propose using ‘microscopic engineering vehicles’ in pesticides to mitigate biotic and abiotic stresses on plants. We discuss their customization and potential mode of action, which may help in addressing the crises caused by stress combination.

Naringin and chloroquine combination mitigates chloroquine-resistant parasite-induced malaria pathogenesis by attenuating the inflammatory response

BackgroundMalaria, characterised by inflammation and multi-organ complications, needs novel chemotherapeutics due to the rise of drug-resistant malaria parasites, which is a serious health issue. Naringin (NGN), a flavanone glycoside (naringenin 7-O-neohesperidose), has a broad spectrum of pharmacological activities but its effect against malaria, alone and in combination, was not deeply investigated. PurposeTo assess the pharmacological efficacy of NGN alone and in combination with chloroquine (CQ) against a Plasmodium strain resistant to CQ and to elucidate its potential mode of action. MethodsThe anti-inflammatory potential of NGN was assessed in mouse microglial cells stimulated with hemozoin by analyzing inflammatory cytokines production. The anti-plasmodial potential of NGN was subsequently tested alone and in combination with CQ against the K1 strain of Plasmodium using the fixed ratio combination method. Further, we evaluated NGN's antimalarial efficacy against the CQ-resistant Plasmodium yoelii nigeriensis N67 strain (P. yoelii), both alone and in combination with CQ, by measuring parasitemia and survival rates. To comprehend the impact of NGN on malaria-induced inflammation in mice, we measured pro-inflammatory cytokines elevated by activated NF-кB signalling. These findings were supported by mRNA and immunohistochemical analyses of malaria-infected mice's liver and brain tissues. ResultsOur study demonstrated that NGN displayed anti-plasmodial activity, which was further augmented when combined with CQ. At 50 µM, NGN significantly reduced the elevation of pro-inflammatory cytokines in synthetic hemozoin-stimulated microglial cells. Compared to P. yoelii-infected mice, NGN (12.5 mg kg-1) significantly reduced parasitemia in mice, resulting in a survival period of up to 13 days. Survival improved by up to 20 days when NGN and CQ were given in combination. NGN, as revealed by immunohistochemical examination of brain and liver tissues, interfered with the NF-кB pathway, potentially reducing the elevation of pro-inflammatory cytokines (TNF-α, IL-1β, IL-18, IFN-γ, and IL-6). This was supported by the overexpression of inflammation-regulatory genes (TGFβ, Nrf2, HO-1, and iNOS) and the downregulation of inflammation-stimulating genes (NF-κB, NLRP3, and caspase-1). Histopathological analysis demonstrated the potential of NGN to restore liver and brain tissues to normal. The substantial decrease in the expression and production of ICAM-1 protein in the brain tissue implies the beneficial effects of NGN, pointing towards its potential for mitigating brain pathology. ConclusionThe findings of this study revealed NGN as a promising drug-like candidate for the management of CQ-resistant parasite-induced malaria pathogenesis for adjunctive therapy in combination with standard antimalarial drugs through its modulation of the NF-κB-mediated inflammation.

Small GTPase PvARFR2 interacts with cytosolic ABA receptor kinase 3 to enhance alkali tolerance in switchgrass.

Soil alkalization has become a serious problem that limits plant growth through osmotic stress, ionic imbalance, and oxidative stress. Understanding how plants resist alkali stress has practical implications for alkaline-land utilization. In this study, we identified a small GTPase, PvARFR2 (ADP ribosylation factors related 2), that positively regulates alkali tolerance in switchgrass (Panicum virgatum) and uncovered its potential mode of action. Overexpressing PvARFR2 in switchgrass and Arabidopsis (Arabidopsis thaliana) conferred transformant tolerance to alkali stress, demonstrated by alleviated leaf wilting, less oxidative injury, and a lower Na+/K+ ratio under alkali conditions. Conversely, switchgrass PvARFR2-RNAi and its homolog mutant atgb1 in Arabidopsis displayed alkali sensitives. Transcriptome sequencing analysis showed that cytosolic abscisic acid (ABA) receptor kinase PvCARK3 transcript levels were higher in PvARFR2 overexpression lines compared to the controls and were strongly induced by alkali treatment in shoots and roots. Phenotyping analysis revealed that PvCARK3-OE × atgb1 lines were sensitive to alkali similar to the Arabidopsis atgb1 mutant, indicating that PvARFR2/AtGB1 functions in the same pathway as PvCARK3 under alkaline stress conditions. Application of ABA on PvARFR2-OE and PvCARK3-OE switchgrass transformants resulted in ABA sensitivity. Moreover, we determined that PvARFR2 physically interacts with PvCARK3 in vitro and in vivo. Our results indicate that a small GTPase, PvARFR2, positively responds to alkali stress by interacting with the cytosolic ABA receptor kinase PvCARK3, connecting the alkaline stress response to ABA signaling.

Circ_BBS9 as an early diagnostic biomarker for lung adenocarcinoma: direct interaction with IFIT3 in the modulation of tumor immune microenvironment.

Introduction: Circular RNAs (circRNAs) have been identified as significant contributors to the development and advancement of cancer. The objective of this study was to examine the expression and clinical implications of circRNA circ_BBS9 in lung adenocarcinoma (LUAD), as well as its potential modes of action. The expression of Circ_BBS9 was examined in tissues and cell lines of LUAD through the utilization of microarray profiling, quantitative real-time polymerase chain reaction (qRT-PCR), and western blot analysis. In this study, we assessed the impact of circ_BBS9 on the proliferation of LUAD cells, as well as its influence on ferroptosis and tumor formation. To analyze these effects, we employed CCK-8 assays and ferroptosis assays. The identification of proteins that interact with Circ_BBS9 was achieved through the utilization of RNA pull-down and mass spectrometry techniques. A putative regulatory network comprising circ_BBS9, miR-7150, and IFIT3 was established using bioinformatics study. The investigation also encompassed the examination of the correlation between the expression of IFIT3 and the invasion of immune cells. Circ_BBS9 was significantly downregulated in LUAD tissues and cell lines. Low circ_BBS9 expression correlated with poor prognosis. Functional experiments showed that circ_BBS9 overexpression inhibited LUAD cell proliferation and promoted ferroptosis in vitro and suppressed tumor growth in vivo. Mechanistically, circ_BBS9 was found to directly interact with IFIT3 and regulate its expression by acting as a sponge for miR-7150. Additionally, IFIT3 expression correlated positively with immune infiltration in LUAD. Circ_BBS9 has been identified as a tumor suppressor in lung adenocarcinoma (LUAD) and holds promise as a diagnostic biomarker. The potential mechanism of action involves the modulation of ferroptosis and the immunological microenvironment through direct interaction with IFIT3 and competitive binding to miR-7150. The aforementioned findings offer new perspectives on the pathophysiology of LUAD and highlight circ_BBS9 as a potentially valuable target for therapeutic interventions.

Bioprospecting of microbial agents and their metabolites as potential inhibitors of Phytophthora cinnamomi, the causal agent of avocado root rot

Phytophthora Root Rot (PRR) is one of the economically important diseases infecting avocado causing substantial yield losses worldwide. In this study, the root rot pathogen infecting avocado has been characterized and potential bioagents targeting PRR have been identified through molecular approaches. Field surveys revealed varying degrees of disease severity (54 %–17.5 %) and incidence (50 %–10 %) in Western Ghats of Tamil Nadu, India. The pathogen was isolated and identified as Phytophthora spp. based on morphology which depicted typical coralloid-type growth patterns. Besides, phylogenetic analysis with the ITS, ras-related protein and COX genes showed closed similarity with P. cinnamomi. The isolates showed significant variation in pathogenicity, with PC(TKI)-10 exhibiting the highest severity (70 %) developing typical symptoms. In vitro efficacy assays through dual-plate and volatiles revealed potential inhibitory effects of Trichoderma harzianum T (MP)-7 (81.11 %) and Bacillus amyloliquefaciens B (TI)-3 (76.67 %) against P. cinnamomi, and Scanning Electron Microscopy (SEM) provided insights into the antagonistic mechanisms, revealing physical interactions and enzymatic lysis of the pathogen by T (MP)-7 and B (TI)-3. Furthermore, Gas Chromatography-Mass Spectrometry (GC-MS) analysis identified bioactive compounds in the crude extracts of these bioagents, elucidating their metabolic pathways and potential modes of action against P. cinnamomi. Our findings highlights the importance of P. cinnamomi in avocado and the mechanism of fungal and bacterial bioagents in the sustainable management of the disease.

A novel technique for inducing ripening and controlling fungal growth in green bananas using warm tangerine oil vapor and its possible modes of action

The study aimed to develop a novel technique for inducing uniform ripening and preventing fungal growth in green bananas, thereby producing high-quality raw materials for various banana-derived products and reducing industrial fruit losses. Tangerine oil (TO) vapor at concentrations of 100, 200, and 400 μl was introduced into a 1-L chamber containing green bananas. The TO was then heated to 50 °C using a heat pad for 10 min, allowing the vapor to release inside the chamber. The chamber with green bananas was held at 30 °C to allow contact with the TO vapor for 12 h. Subsequently, the bananas were kept at 30 ± 2 °C for 10 days to ripen. The experiment was compared to acetylene and control treatments. TO was also tested to inhibit the growth of Lasiodiplodia theobromae (the cause of crown rot disease in bananas) during storage, and the assessment of banana ripening quality was conducted. The potential mode of action in both ripening acceleration and fungal control was investigated. Results showed that bananas treated with TO vapor at 200 μl and acetylene exhibited an earlier climacteric rise within five days compared to the control (10 days). TO vapor (≥200 μl) demonstrated efficacy against L. theobromae on agar and on the banana surface. TO (200 μl) contributed to improved banana ripening quality, as evidenced by reduced firmness and pH, increased total soluble solids (TSS) and titratable acidity (TA) content, and the absence of mold disease on the surface after ten days. Regarding the mode of action, the peel morphology of bananas treated with TO revealed a large pore size during ripening, affirming the efficiency of TO in accelerating banana ripening, contributing to the acceleration of chlorophyll reduction and the transition to a yellow banana. Limonene, a key component, facilitated cell wall penetration, causing cell membrane leakage in fungi. This finding confirms that implementing this technique for controlling banana ripening in the industry could lead to superior-quality bananas with various beneficial product applications.

Antimicrobial Activity and Mechanisms of Punicalagin against Vibrio parahaemolyticus.

This study sought to explore the antimicrobial activity of punicalagin against V. parahaemolyticus and its potential modes of action. V. parahaemolyticus ATCC 17802 and RIMD 2210633Sm were exposed to punicalagin, and the energy production, membrane potential, and envelope permeability, as well as the interaction with cell biomolecules, were measured using a variety of fluorescent probes combined with electrophoresis and Raman spectroscopy. Punicalagin treatment disrupted the envelope integrity and induced a decrease in intracellular ATP and pH. The uptake of 1-N-phenyl-naphtylamine (NPN) demonstrated that punicalagin weakened the outer membrane. Punicalagin damaged the cytoplasmic membrane, as indicated by the membrane depolarization and the leakage of intracellular potassium ions, proteins, and nucleic acids. Electronic microscopy observation visualized the cell damage caused by punicalagin. Further, gel electrophoresis coupled with the Raman spectrum assay revealed that punicalagin affected the protein expression of V. parahaemolyticus, and there was no effect on the integrity of genomic DNA. Therefore, the cell envelope and proteins of V. parahaemolyticus were the assailable targets of punicalagin treatment. These findings suggested that punicalagin may be promising as a natural bacteriostatic agent to control the growth of V. parahaemolyticus.

Antimicrobial activity of rambutan peel colouring agent containing cardamom oil in shrimp paste and its potential mode of action

SummaryRambutan peel (RP), an agricultural by‐product from rambutan trading, contains the red pigment anthocyanin. This research explores developing a colouring agent from RP, utilising cardamom oil (CM) to enhance colour stability, improve antimicrobial functionality, and create a new high‐value red colouring agent from fruit waste. A colouring agent was produced by placing 50 g of RP inside a 1 L glass box, and varying amounts of CM (5 μL, 25 μL, 50 μL, and 100 μL) on filter paper were placed inside the box lid. The box was sealed and left closed for 8 h to allow CM vapour to release onto the surface of RP before being dried at 30 °C and ground into powder. The antibacterial impact of CM‐treated RP on Staphylococcus aureus was assessed using the nutrient broth. The CM‐treated RP powder was used as a food colouring agent in shrimp paste to evaluate the shelf life of shrimp paste during storage at 30 °C. Results revealed that treated RP with 25 μL of CM showed the best red colour appearance in the dried powder. RP with 25 μL of CM exhibited time killing against S. aureus (approximately 4 log10cfu g−1) after being added to the medium for 6 h, disrupting the cell membrane integrity of S. aureus and leading to cell lysis. RP treated with 25 μL of CM also prevented darkening in shrimp paste, maintaining higher values for L* and a*, resulting in increased lightness and a more red appearance. It also inhibited pathogenic bacteria, extending shelf life, and preserving colour stability for 14 days, compared to the control's 7 days. These insights provide valuable guidance for the sustainable utilisation of agricultural waste in the food colouring industry.

Exploring the modulatory influence on the antimalarial activity of amodiaquine using scaffold hybridisation with ferrocene integration

Amodiaquine (AQ) is a potent antimalarial drug used in combination with artesunate as part of artemisinin-based combination therapies (ACTs) for malarial treatment. Due to the rising emergence of resistant malaria parasites, some of which have been reported for ACT, the usefulness of AQ as an efficacious therapeutic drug is threatened. Employing the organometallic hybridisation approach, which has been shown to restore the antimalarial activity of chloroquine in the form of an organometallic hybrid clinical candidate ferroquine (FQ), the present study utilises this strategy to modulate the biological performance of AQ by incorporating ferrocene. Presently, we have conceptualised ferrocenyl AQ derivatives and have developed facile, practical routes for their synthesis. A tailored library of AQ derivatives was assembled and their antimalarial activity evaluated against chemosensitive (NF54) and multidrug-resistant (K1) strains of the malaria parasite, Plasmodium falciparum. The compounds generally showed enhanced or comparable activities to those of the reference clinical drugs chloroquine and AQ, against both strains, with higher selectivity for the sensitive phenotype, mostly in the double-digit nanomolar IC50 range. Moreover, representative compounds from this series show the potential to block malaria transmission by inhibiting the growth of stage II/III and V gametocytes in vitro. Preliminary mechanistic insights also revealed hemozoin inhibition as a potential mode of action.

Exogenous hydrogen sulphide promotes plant flowering through the Arabidopsis splicing factor AtU2AF65a.

Alternative splicing (AS) is an important regulatory mode at the post-transcriptional level, through which many flowering genes regulate floral transition by producing multiple transcripts, and splicing factors have essential roles in this process. Hydrogen sulphide (H2S) is a newly found gasotransmitter that has critical physiological roles in plants, and one of its potential modes of action is via persulfidation of target proteins at specific cysteine sites. Previously, it has been shown that both the splicing factor AtU2AF65a and H2S are involved in the regulation of plant flowering. This study found that, in Arabidopsis, the promoting effect of H2S on flowering was abolished in atu2af65a-4 mutants. Transcriptome analyses showed that when AtU2AF65a contained mutations, the regulatory function of H2S during the AS of many flowering genes (including SPA1, LUH, LUG and MAF3) was inhibited. The persulfidation assay showed that AtU2AF65a can be persulfidated by H2S, and the RNA immunoprecipitation data indicated that H2S could alter the binding affinity of AtU2AF65a to the precursor messenger RNA of the above-mentioned flowering genes. Overall, our results suggest that H2S may regulate the AS of flowering-related genes through persulfidation of splicing factor AtU2AF65a and thus lead to early flowering in plants.

Review and Perspectives on Bifidobacterium lactis for Infants' and Children's Health.

The influence of microbiota dysbiosis in early life is increasingly recognized as a risk factor for the development of several chronic diseases later in life, including an increased risk of asthma, eczema, allergies, obesity, and neurodevelopmental disorders. The question whether the potential lifelong consequences of early life dysbiosis could be mitigated by restoring microbiota composition remains unresolved. However, the current evidence base suggests that protecting the normal development of the microbiome during this critical developmental window could represent a valuable public health strategy to curb the incidence of chronic and lifestyle-related diseases. Probiotic Bifidobacteria are likely candidates for this purpose in newborns and infants considering the natural dominance of this genus on microbiota composition in early life. Moreover, the most frequently reported microbiota composition alteration in association with newborn and infant diseases, including necrotizing enterocolitis and diarrhea, is a reduction in Bifidobacteria levels. Several studies have assessed the effects of B. animalis subsp. lactis strains in newborns and infants, but recent expert opinions recommend analyzing their efficacy at the strain-specific level. Hence, using the B94 strain as an example, this review summarizes the clinical evidence available in infants and children in various indications, discussing the safety and potential modes of actions while providing perspectives on the concept of "non-infant-type" probiotics for infants' health.

Maternal impacts of pre-conceptional glyphosate exposure

Maternal glyphosate (GLY) impacts remain unclear despite associations between urinary GLY and birth outcomes. Whether maternal pre-conceptional GLY exposure would have phenotypic and molecular impacts in the dam and offspring was tested. Female C57BL6 mice (6 wk) were exposed to saline (CT; n = 20) or GLY (2 mg/kg; n = 20) per os five d per week for 20 wk. Females were housed with males and on gestation day (GD) 14, divided into: CT non-pregnant (CNP), CT pregnant (CP), GLY non-pregnant (GNP), GLY pregnant (GP). Another cohort (CT; n = 10 or GLY; n = 10) completed three pregnancy rounds and pregnancy index (PI), number of pups per litter and pups surviving to postnatal day (PND) 5 calculated. The PI in GLY mice was higher in breeding rounds 1 and 2, but lower in round 3. Pregnancy increased (P ≤ 0.1) GD14 liver and ovary weight. Spleen weight was increased (P < 0.05) in GP relative to GNP mice. No offspring phenotypic impacts were observed. Approximately six months after cessation of exposure, secondary follicle number was reduced (P < 0.05) by pre-conceptional GLY exposure. The ovarian proteome analyzed by LC-MS/MS was altered (P < 0.05) by pregnancy (49 increased, 43 decreased) and GLY exposure (non-pregnant: 75 increased, 22 decreased, pregnant: 27 increased, 29 decreased; aged dams: 60 increased, 98 decreased) with several histone proteins being altered. These findings support ovarian transient and persistent impacts of GLY exposure and identify pathways as potential modes of action.

Hydronidone ameliorates liver fibrosis by inhibiting activation of hepatic stellate cells via Smad7-mediated degradation of TGFβRI.

Liver fibrosis is a wound-healing reaction that eventually leads to cirrhosis. Hydronidone is a new pyridine derivative with the potential to treat liver fibrosis. In this study, we explored the antifibrotic effects of hydronidone and its potential mode of action. The anti-hepatic fibrosis effects of hydronidone were studied in carbon tetrachloride (CCl4 )- and 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC)- induced animal liver fibrosis. The antifibrotic mechanisms of hydronidone were investigated in hepatic stellate cells (HSCs). The antifibrotic effect of hydronidone was further tested after Smad7 knockdown in HSCs in mouse models of fibrosis. In animal models, hydronidone attenuated liver damage and collagen accumulation, and reduced the expression of fibrosis-related genes. Hydronidone decreased the expression of fibrotic genes in HSCs. Impressively, hydronidone significantly upregulated Smad7 expression and promoted the degradation of transforming growth factor β receptor I (TGFβRI) in HSCs and thus inhibited the TGFβ-Smad signalling pathway. Specific knockdown of Smad7 in HSCs in vivo blocked the antifibrotic effect of hydronidone. Hydronidone ameliorates liver fibrosis by inhibiting HSCs activation via Smad7-mediated TGFβRI degradation. Hydronidone is a potential drug candidate for the treatment of liver fibrosis.

Biological Control Mechanisms of Bacillus cabrialesii subsp. tritici TSO2T against Fusarium languescens, the Causal Agent of Wilt in Jalapeño Peppers

Jalapeño peppers (Capsicum annuum var. Jalapeño) represent one of the most important crops in Mexico. However, many plant diseases, such as wilt caused by strains of the genus Fusarium, reduce its yield. A sustainable alternative to control diseases is the use of biological control agents (BCAs), for example, beneficial microorganisms such as strains of the genus Bacillus. This study aims to analyze the potential use of B. cabrialesii subsp. tritici TSO2T as a BCA and elucidate its potential modes of action against Fusarium strains causing wilt in Jalapeño peppers. For this, symptomatic samples were collected in a commercial field in the Yaqui Valley, Mexico. Six Fusarium isolates were morphologically and molecularly characterized. After pathogenicity tests, F. languescens CE2 was found to be pathogenic. In screening assays for biocontrol bacteria, strain TSO2T, which was isolated from soil in a wheat commercial field under an organic production system and preserved in the Culture Collection of Native Soil and Endophytic Microorganisms (COLMENA), had the best biocontrol effect against CE2, and its cell-free filtrate reduced mycelial growth by 30.95%. Genome mining (antiSMASH) of strain TSO2T allows us to identify gene clusters associated with biocontrol, such as fengycin, surfactin, bacillibactin, bacilysin, bacillaene, subtilosin A, and sporulation killing factor, which codify to antimicrobial metabolites and are associated with swarming motility of the studied beneficial strain. These results demonstrate the effectiveness of B. cabrialesii subsp. tritici TSO2T as a potential BCA for the control of Fusarium wilt, through competition and a complex of antifungal metabolites, which still need to be validated through metabolomic analysis. This study highlights the importance of the use of integrative genomic and bioactivity-guided methodologies in understanding biocontrol pathosystems.

Mode of action of antimicrobial agents albofungins in eradicating penicillin- and cephalosporin-resistant Vibrio parahaemolyticus biofilm.

Albofungin is a promising broad-spectrum antimicrobial compound against multidrug-resistant bacteria. In the present study, we further investigated albofungin's biofilm eradication activity and its potential mode of action against drug-resistant Vibrio parahaemolyticus. Among all derivatives, albofungin exhibited the best antibiofilm and antibacterial activity with rapid killing effects at 0.12 µg mL-1. Confocal microscopy observation exhibited that albofungin disrupted V. parahaemolyticus biofilms by killing or dispersing biofilm cells. Meanwhile, scanning electron microscope and fluorescent staining experiments demonstrated that albofungin rapidly destroyed the integrity and permeability of the bacterial cell membrane. Moreover, this study revealed an antibiofilm mechanism of albofungin involving inhibition of peptidoglycan biosynthesis, flagella assembly pathways, and secretion system proteins in V. parahaemolyticus by quantitative proteomics and validation experiments. Our results highlighted albofungin's mechanism of action in planktonic cells and biofilms and suggested further development and potential applications of albofungin for treating infections caused by penicillins-and-cephalosporins-resistant V. parahaemolyticus. IMPORTANCE Infections caused by multidrug-resistant bacteria, as well as a scarcity of new antibiotics, have become a major health threat worldwide. To tackle the demand for new and effective treatments, we investigated the mechanism of action of albofungin, a natural product derived from Streptomyces, which exhibits potent antimicrobial activity against multidrug-resistant bacteria. Albofungin showed potent biofilm eradication activity against penicillins-and-cephalosporins-resistant Vibrio parahaemolyticus, which expresses a novel metallo-β-lactamase and, thus, reduces their sensitivity to various antibiotics. We observed membrane disruption and permeation mechanisms in planktonic cells and biofilms after albofungin treatment, while albofungin had a weak interaction with bacterial DNA. Moreover, the antibiofilm mechanism of albofungin included inhibition of peptidoglycan biosynthesis, flagellar assembly pathways, and secretion system proteins. Our finding suggested potential applications of albofungin as an antibacterial and antibiofilm therapeutic agent.