We describe the androgen receptor (AR) agonistic/antagonistic effects of 140 veterinary drugs regulated in Republic of Korea, by setting maximum residue limits. It was conducted using two in vitro test guidelines of the Organization for Economic Cooperation and Development (OECD)—the AR-EcoScreen AR transactivation (TA) assay and the 22Rv1/MMTV_GR-KO AR TA assay. These were performed alongside the AR binding affinity assay to confirm whether their AR agonistic/antagonistic effects are based on the binding affinity to AR. Prior to conducting the AR TA assay, the proficiency test was passed the proficiency performance criterion for the AR agonist and AR antagonist assays. Among the veterinary drugs tested, four veterinary drugs (dexamethasone, trenbolone, altrenogest, and nandrolone) and six veterinary drugs (cymiazole, dexamethasone, zeranol, phenothiazine, bromopropylate, and isoeugenol) were determined as AR agonist and AR antagonist, respectively in both in vitro AR TA assays. Zeranol exhibited weak AR agonistic effects with a PC₁₀ value only in the 22Rv1/MMTV_GR-KO AR TA assay. Regarding changing the AR agonistic/antagonistic effects through metabolism, the AR antagonistic activities of zeranol, phenothiazine, and isoeugenol decreased significantly in the presence of phase I + II enzymes.These data indicate that various veterinary drugs could have the potential to disrupt AR-mediated human endocrine system. Furthermore, this is the first report providing information on AR agonistic/antagonistic effects of veterinary drugs using in vitro OECD AR TA assays.

C₆₀ fullerene (C₆₀) is a nano-pollutant that can damage the respiratory system. Eugenol exhibits significant anti-inflammatory and antioxidant properties. We aimed to investigate the time course of C₆₀ emulsion-induced pulmonary and spermatic harms, as well as the effect of eugenol on C₆₀ emulsion toxicity. The first group of mice (protocol 1) received intratracheally C₆₀ emulsion (1.0 mg/kg BW) or vehicle and were tested at 12, 24, 72 and 96 h (F groups) thereafter. The second group of mice (protocol 2) received intratracheally C₆₀ emulsion or vehicle, 1 h later were gavaged with eugenol (150 mg/kg) or vehicle, and experiments were done 24 h after instillation. Lung mechanics, morphology, redox markers, cytokines and epididymal spermatozoa were analyzed. Protocol 1: Tissue damping (G) and elastance (H) were significantly higher in F24 than in others groups, except for H in F72. Morphological and inflammatory parameters were worst at 24 h and subsequently declined until 96 h, whereas redox and spermatic parameters worsened over the whole period. Eugenol eliminated the increase in G, H, cellularity, and cytokines, attenuated oxidative stress induced by C60 exposure, but had no effect on sperm. Hence, exposure to C₆₀ emulsion deteriorated lung morphofunctional, redox and inflammatory characteristics and increased the risk of infertility. Furthermore, eugenol avoided those changes, but did not prevent sperm damage.

This study aimed to evaluate possible synergistic interactions on antimicrobial and antioxidant efficacy of clove and cinnamon oil components in combination and characterization of compounds responsible for synergistic interactions using TLC bioautography followed by checkerboard titration, isobologram analysis, and spectrometric characterization. Among the combinations tested, cinnamaldehyde from cinnamon oil and eugenol from clove oil in combination showed a synergistic antimicrobial interaction against foodborne microbes Listeria monocytogenes (fractional inhibitory concentration index (FICI): 0.31), Salmonella typhimurium (FICI: 0.41), and Aspergillus niger (FICI: 0.48), and synergistic antioxidant efficacy (combination index: 0.78) in in vitro model. Cinnamaldehyde/eugenol blend did not show any cytotoxic effect (IC₅₀ > 1000 μg/ml) in human normal keratinocyte cell line. The results provide evidence that the cinnamaldehyde/eugenol blend may help in designing a more potent novel natural antimicrobial and antioxidant agent in food and pharmaceutical industries.

The novelty of the present research is conducting a new method in the systemic resistance of plant diseases by using distinct marine extracts. The ability of two octopus extracts to reduce the wilt disease caused by Fusarium oxysporum was observed. The applied methods are soaked roots (SR) and foliar shoots (FS). The antioxidant enzyme activities, percent disease index (PDI), and growth parameters were measured. In vitro antifungal potential of the octopus extracts against F. oxysporum was examined. The obtained result shows that SR extracts reduced PDI. Additionally, all the tested treatments promoted the growth and photosynthetic pigments of the infected plants. SR (in ethanolic extracts) was the most prominent inducer which offered a high advancement in the total soluble protein contents. Also, SR (in methanolic extracts) was the most suitable inducer which provided a very necessary development not only in the total phenol but also in the peroxidase (POD) and polyphenol oxidase (PPO) activities. GC-MS investigation of the octopus extracts exhibited that the compounds which possess antifungal activity were furoscrobiculin B and/or eugenol. They demonstrated a notable antifungal potential against F. oxysporum with a maximum activity of 38.5 and 12.7 mm ZOI after the treatment with the ethanolic and methanolic extract, respectively. FTIR results illustrated the functional group of the compound responsible for the antifungal activity. Additionally, an atomic absorption result reveals that there are traces of metals detected such as Pb, Ag, Cu, Zn, and Mg. The antifungal activity was decreased as the concentrations were reduced. Accordingly, the present extracts may be used as the vital agents in the agricultural field to restrain the plant pathogenic fungi, especially F. oxysporum from a proliferation.

Biological methods offer eco-friendly and cost-effective alternatives for the synthesis of silver nanoparticles (AgNPs). The present study highlights a green process where AgNPs were synthesized and optimized by using silver nitrate (AgNO₃) and the aqueous extract of Piper betle (Pbet) leaf as the reducing and capping agent. The stable and optimized process for the synthesis of Pbet-AgNPs was exposure of reaction mixture into the sunlight for 40 min, pH 9.0, and 2 mM AgNO₃ using 1:4 diluted Pbet leaf aqueous extract. The optimized Pbet-AgNPs were characterized by UV–visible spectroscopy, high-resolution field emission scanning electron microscopy (FE-SEM), X-ray diffractometry (XRD), and Fourier-transform infrared spectroscopy (FTIR). The prepared Pbet-AgNPs were spherical in shape with size in the range of 6–14 nm. These nanoparticles were stable for 6 months in aqueous solution at room temperature under dark conditions. The biogenic synthesized Pbet-AgNPs are found to have significant antifungal activity against plant pathogenic fungi, Alternaria brassicae and Fusarium solani. Synthesized Pbet-AgNPs potentially reduced the fungal growth in a dose-dependent manner. Microscopic observation of treated mycelium showed that Pbet-AgNPs could disrupt the mycelium cell wall and induce cellular permeability. Protein leakage assay supports these findings. Overall, this study revealed the efficacy of green synthesized AgNPs to control the plant fungal pathogens. Pbet leaves are a rich source of phenolic biomolecule(s). It was hypothesized that these biomolecule(s) mediated metal reduction reactions. In this context, the present work investigates the phytobiomolecule(s) of the aqueous extract of Pbet leaves using high-resolution liquid chromatography–mass spectroscopy (HR-LCMS) method. The analysis revealed that eugenol, chavicol, and hydroxychavicol were present in the Pbet aqueous extract.

Aging represents the accumulation of progressive changes in a human being over time and can cover physical, psychological, and social changes. It is an oxidative stress-associated process that progresses with age. The antioxidant activity of either eugenol (EU) or carvacrol (CAR) for aging in rats induced by D-gal for 42 days was investigated in the current study using 10 and 20 mg of EU/kg/day/orally, while CAR was supplemented by 40 and 80 mg /kg/day/orally. Biochemical, mRNA expression, and histopathological assessments of brain samples evaluated the oxidative alterations induced by D-gal and the protective role of EU and CAR. Results showed that D-gal was causing oxidative alternation of the brain that was recognized via upregulation of p53 and p21 mRNA expression levels, as aging markers and Bax mRNA expression level, as an apoptotic marker. Also, the results observed alterations in the levels of biochemical markers as creatine phosphokinase (CPK) and triacylglycerol (TAG), besides, enhancement of brain antioxidant capacity. Finally, these results compared with the groups treated with EU and CAR to observe that the EU and CAR potentially attenuate these aging-related oxidative alterations in a dose-dependent manner. Finally, we can conclude that EU and CAR supplementations are considered promising natural protective compounds that could delay aging and maintain health.

This study aims to determine the maximum concentration and the long-term effects after exposure to a bio-insecticide with active ingredients eugenol and azadirachtin on the survival rate, immunity, and growth of Nile tilapia. The method used in this study was experimental, using a completely randomized design (CRD) with six treatments and three replications. Fishes were exposed to eugenol and azadirachtin at concentrations 10, 20, 30, 40, and 50% of LC50 value for 14 days, followed by 14 days of maintenance to see the effect on growth. The results showed that 66 mg.L-1 treatment was a concentration that did not interfere with the survival rate of Nile tilapia, which was 86.7%. The number of leukocytes increased on the third day by the highest increase in 66 mg.L-1 treatment at 12.01 × 104 cells.mm-3. Meanwhile, erythrocytes decreased, with the highest decrease in 66 mg.L-1 treatment at 1.13 × 106 cells.mm-3. The average growth rate in fish slowed down with increasing concentrations of exposure, with the lowest average growth in length and absolute weight in the 66 mg.L-1 treatment was 0.57 cm and 1.68 g.

Titanium dioxide nanoparticles (TiO₂ NPs) are widely used in food, edible dyes, and other commercial products. Human exposure to TiO₂ NPs has raised concerns regarding their toxic potential. Various studies have evaluated the TiO₂ NPs-induced toxicity, oxidative damage to the cellular components, and genotoxicity. In the present study, we examined whether co-treatment with the dietary antioxidant eugenol can attenuate or protect against TiO₂ NPs-induced toxicity. We exposed the adult male Wistar rats to TiO₂ NPs (150 mg/kg body weight) by intraperitoneal injection (i.p.) either alone or as co-treatment with eugenol (1-10 mg/kg body weight) once a day for 14 days. The untreated rats were supplied saline and served as control. Titanium (Ti) accumulation in various tissues was analyzed by inductively coupled plasma mass spectrometry. Serum levels of liver and kidney biomarkers and oxidative stress markers in the liver, kidney, and spleen were determined. A significant increase in hydrogen peroxide level confirmed that oxidative stress occurred in these tissues. TiO₂ NPs induced oxidation of lipids, and decreased glutathione level and antioxidant enzyme activity in the kidney, liver, and spleen of treated rats. TiO₂ NPs also increased the serum levels of alanine aminotransferase, alkaline phosphatase, aspartate aminotransferase, albumin, and total cholesterol and decreased the blood urea nitrogen, uric acid, and total bilirubin in serum, which indicates oxidative damage to the liver and kidney. In eugenol and TiO₂ NPs co-treated rats, all these changes were mitigated. Single-cell gel electrophoresis (comet assay) of lymphocytes showed longer comet tail length in TiO₂ NPs-treated groups, indicating DNA damage while tail length was reduced in eugenol and TiO₂ NPs co-treated groups. Thus, it seems that eugenol can be used as a chemoprotective agent against TiO₂ NPs-induced toxicity.

Plutella xylostella L. is a cosmopolitan pest of wild and cultivated crucifer vegetables worldwide. It has developed resistance to almost all commercial chemicals, making them one of the most problematic field pests in China. The natural plant extracts and essential oils (EOs) could be a safe alternative for agricultural pests. The development and production of EOs decrease the negative effects of synthetic chemicals. In the present study, the fumigation activity of 8 pure monoterpenes against P. xylostella was evaluated. Results from fumigation tests revealed that 8 tested compounds exhibited various degrees of toxicity against adults of the diamondback moth. Cuminaldehyde was the most toxic compound based on the 12-h LC₅₀ (0.17 mg/L) and 24-h LC₅₀ (0.12 mg/L) values, respectively. Also for larvae and eggs, cuminaldehyde was the most toxic compound. The 12-h LC₅₀ value for cuminaldehyde to 1st, 2nd, and 3rd instar larvae was 0.10 mg/L, 0.12 mg/L, and 0.55 mg/L, respectively. The 24-h LC₅₀ value for the different instar larvae was 0.07 mg/L, 0.09 mg/L, and 0.35 mg/L, respectively. The 24-h LC₅₀ value for eggs (endpoint hatching rate) was 1.95 mg/L for cuminaldehyde, followed by carvacrol and eugenol (2.05 mg/L and 2.31 mg/L, respectively). Cuminaldehyde was very friendly to the larvae and adults of Harmonia axyridis and did not cause any mortality. Our results indicated that cuminaldehyde had potential insecticidal activity against P. xylostella and could be utilized in the novel biological pesticide development.

Cleome amblyocarpa Barr. and Murb is a medicinal plant widespread in North Africa and widely used in Tunisia to treat diabetes and colic. The non-volatile (polyphenols, flavonoids, tannins, and flavonols) and volatile compounds (GC-MS) of C. amblyocarpa leaves and stems have been studied. The antioxidant, antimicrobial, analgesic, and cytotoxic activities of hydroalcoholic extracts of C. amblyocarpa leaves and stems were also investigated. The major volatile components were β-caryophyllene (46.9%), eugenol (25.6%), ethyl 3-methylpentanoate (16.2%), 7-epi-silphiperfol-5-ene (11.0%), and α-copaene (7.0%). The antioxidant activity has been evaluated using various in vitro assays, such as DPPH free radical scavenging activity, iron-chelating capability, and ability to inhibit lipid peroxidation (TBARS). The antibacterial and antifungal effectiveness of leaves and stems parts of Cleome amblyocarpa were investigated by means of the disc diffusion and microdilution techniques. The in vitro cytotoxicity of the hydroalcoholic extract of C. amblyocarpa on A549 and H1299 lung adenocarcinoma cells was determined using the crystal violet assay. The acute toxicity of the extracts on Swiss albino mice at the doses of 3000, 1500, and 500 mg/kg body weight was evaluated. The analgesic effect of leaves and stems extract was also determined by means of the acetic acid induced writhing test. The results indicated that the leaves have higher phenols, and flavonoids contents and potential antioxidant, antimicrobial, and anticancer activities in comparison to stem. In addition, the aerial part of C. amblyocarpa did not cause signs of toxicity or death in animals at doses up to 3000 mg/kg and have a significant analgesic activity.