Investigating the Impact of First-Line Anti-Tuberculosis Drugs Encapsulated in a Eugenol-Based Nanoemulsion on Human Serum Albumin.

BACKGROUND: Eugenol exhibits broad-spectrum antibacterial and anti-inflammatory properties. However, cytotoxicity at high concentrations limits the full utilization of eugenol-based drug complexes. Formulations of multidrug-loaded eugenol-based nanoemulsions have reduced cytotoxicity; however, it remains crucial to understand how these eugenol complexes interact with primary human carrier proteins to design and develop therapeutic alternatives. Consequently, this study primarily aims to investigate the impact on Human Serum Albumin (HSA) when it interacts with eugenol-based complexes loaded with first-line anti-tuberculosis drugs. METHODS: This study used various spectroscopic such as UV-visible spectroscopy, Fluorescence spectroscopy, Fourier-transform infrared spectroscopy and computational methods such as molecular docking and 100 ns molecular simulation to understand the impact of eugenol-based first-line anti-tuberculosis drug-loaded nanoemulsions on HSA structure. RESULTS: The binding of the HSA protein and eugenol-based complexes was studied using UV-visible spectroscopic analysis. Minor changes in the fluorophores of the protein further confirmed binding upon interaction with the complexes. The Fourier-transform infrared spectra showed no significant changes in protein structure upon interaction with eugenol-based multidrug-loaded nanoemulsions, suggesting that this complex is safe for internal administration. Unlike eugenol or first-line anti-tuberculosis alone, molecular docking revealed the strength of the binding interactions between the complexes and the protein through hydrogen bonds. The docked complexes were subjected to a 100 ns molecular dynamics simulation, which strongly supported the conclusion that the structure and stability of the protein were not compromised by the interaction. CONCLUSIONS: From the results we could comprehend that the eugenol (EUG)-drug complex showed greater stability in HSA protein structure when compared to HSA interacting with isoniazid (INH), rifampicin (RIF), pyrazinamide (PYR), or ethambutol (ETH) alone or with EUG alone. Thus, inferring the potential of EUG-based drug-loaded formulations for a safer and efficient therapeutic use.

Synergistic Effect between Eugenol and 1,8-Cineole on Anesthesia in Guppy Fish (Poecilia reticulata).

This study aimed to evaluate the synergistic effect between eugenol and 1,8-cineole on anesthesia in female guppy fish (Poecilia reticulata). Experiment I evaluated the concentrations of 0, 12.5, 25, 50, and 75 mg/L of eugenol and 0, 100, 200, 300, and 400 mg/L of 1,8-cineole for times of induction and recovery from anesthesia. Experiment II divided fish into 16 study groups, combining eugenol and 1,8-cineole in pairs at varying concentrations, based on the dosage of the chemicals in experiment I. The results of the anesthesia showed that eugenol induced fish anesthesia at concentrations of 50 and 70 mg/L, with durations of 256.5 and 171.5 s, respectively. In contrast, 1,8-cineole did not induce fish anesthesia. In combination, using eugenol at 12.5 mg/L along with 1,8-cineole at 400 mg/L resulted in fish anesthesia at a time of 224.5 s. Increasing the eugenol concentration to 25 mg/L, combined with 1,8-cineole at 300 and 400 mg/L, induced fish anesthesia at times of 259.0 and 230.5 s, respectively. For treatments with eugenol at 50 mg/L combined with 1,8-cineole at 100 to 400 mg/L, fish exhibited anesthesia at times of 189.5, 181.5, 166.0, and 157.5 s. In the case of eugenol at 75 mg/L, fish showed anesthesia at times of 175.5, 156.5, 140.5, and 121.5 s, respectively. The testing results revealed that 1,8-cineole as a single treatment could not induce fish anesthesia. However, when supplementing 1,8-cineole in formulations containing eugenol, fish exhibited a significantly faster induction of anesthesia (p < 0.05). Furthermore, all fish that underwent anesthesia were able to fully recover without any mortality. However, the shorter anesthesia duration resulted in a significantly prolonged recovery time. In conclusion, eugenol and 1,8-cineole work better together as anesthetics than when used separately, and demonstrated the safety of using these anesthetic agents on guppy fish.

Eugenol-Loaded Transethosomal Gel for Improved Skin Delivery and Treatment of Atopic Dermatitis.

Atopic dermatitis is a skin condition characterized by lichenification (thickening and increased skin marking), eczematous lesions, dry skin, itching, and pruritus. Eugenol is an aromatic polyphenolic compound that has attracted the attention of researchers due to its anti-inflammatory, anti-oxidant, and anti-cancer properties. The primary goal of the present study was to develop and evaluate eugenol-loaded transethosomes for the treatment of AD. Eugenol-loaded transethosomes were formulated using the ethanol injection method and subsequently subjected to particle size analysis, zeta potential, entrapment efficiency, deformability index, and HRTEM analysis. Transethosomal gel was prepared by direct-dispersion method by using Carbopol 940®. Results showed transethosomes to be lipid bilayer structures with acceptable size, and high entrapment efficiency. Transethosomal formulation showed shear-thinning behavior. Eugenol-loaded transethosomal gel was significantly able to enhance the retention of the drug in the skin. Transethosomal gel was significantly able to reduce Ear thickness, DLC, TLC, and IL-6 levels in mice model of AD. These results indicate that the eugenol-loaded transethosomal gel could be a promising carrier for the topical administration of eugenol for the treatment of AD.

Pickering nanoemulsion loaded with eugenol contributed to the improvement of konjac glucomannan film performance.

Konjac glucomannan (KGM) is becoming a very potential food packaging material due to its good film-forming properties and stability. However, KGM film has several shortcomings such as low mechanical strength, strong water absorption, and poor self-antibacterial performance, which limits its application. Therefore, in order to enhance the mechanical and functional properties of KGM film, this study prepared Pickering nanoemulsion loaded with eugenol and added it to the KGM matrix to explore the improvement effect of Pickering nanoemulsion on KGM film properties. Compared to pure KGM film and eugenol directly added film, the mechanical strength of Pickering-KGM film was significantly improved due to the establishment of ample hydrogen bonding interactions between the ß-cyclodextrin inclusion complex system and KGM. Pickering-KGM film had significant antioxidant capacity than pure KGM film and eugenol directly added KGM film (eugenol-KGM film) (~3.21 times better than KGM film, ~0.51 times better than eugenol-KGM film). In terms of antibacterial activity, Pickering-KGM film had good inhibitory effect on Escherichia coli, Staphylococcus aureus, and Candida albicans, and raspberry preservation experiment showed that the shelf life of the Pickering-KGM film could be extended to about 6 days. To sum up, this study developed a novel means to improve the film performance and provide a new insight for the development and application of food packaging film.

Exploring promising minor natural phenolic compounds in neuroprotection-related preclinical models.

Neurodegenerative diseases, such as Alzheimer's disease and Parkinson's disease, are characterised by the progressive loss of specific neuronal cell populations due to multifactorial factors, including neurochemical and immunological disturbances. Consequently, patients can develop cognitive, motor and behavioural dysfunctions, which lead to impairments in their quality of life. Over the years, studies have reported on the neuroprotective properties inherent in phenolic compounds. Therefore, this review highlights the most recent scientific findings regarding phenolic compounds as promising neuroprotective molecules against neurodegenerative diseases.

Interaction of eugenol-based anti-tuberculosis nanoemulsion with bovine serum albumin: A spectroscopic study including Rifampicin, Isoniazid, Pyrazinamide, and Ethambutol.

Tuberculosis (TB), a deadly infectious disease, is primarily caused by the bacterium Mycobacterium tuberculosis. The misuse of antibiotics has led to the development of drug resistance, prompting researchers to explore new technologies to combat multidrug-resistant Tuberculosis (MDR TB). Phospholipid-based nanotherapeutics, such as nanoemulsions, are gaining traction as they enhance drug solubility, stability, and bioavailability. Our study focuses on the interaction between Bovine Serum Albumin (BSA) and a drug-loaded nanoemulsion based on Eugenol. This nanoemulsion incorporates Eugenol, Clove, cinnamon oil, and first-line anti-tuberculosis drugs like Rifampicin, Isoniazid, Pyrazinamide, and Ethambutol. The primary objective is to assess the biosafety profile of the nanoemulsion upon interaction with BSA. We employed Fluorescence, UV-visible, and Fourier Transform Infrared Spectroscopy (FTIR) to analyze this interaction. UV-visible spectroscopy detected changes in hydrophobicity due to structural alterations in BSA near the tryptophan residue, leading to the formation of ground-state complexes. Fluorescence spectroscopy demonstrated that the nanoemulsion effectively quenched fluorescence originating from tryptophan and tyrosine residues. Studies using synchronous and three-dimensional spectroscopy point to a potential modification of the aromatic environment of BSA by the nanoemulsion. Resonance light scattering spectra indicated the formation of large aggregates due to the interaction with the nanoemulsion. The second derivative FTIR spectra showed an increase in the magnitude of secondary structure bands, suggesting a conformational shift. This research has significant pharmacological implications for developing safer, more targeted drug delivery systems. The information obtained from the interaction of the nanoemulsion with the blood carrier protein is vital for the future development of superior carriers with minimal adverse effects on patients. It is crucial to remember that conformational changes brought on by drug-ligand complexes attaching to carrier proteins may have negative consequences. Therefore, this study enhances the in vitro evaluation of potential adverse effects of the nanoemulsion on serum proteins.

Prophylactic and therapeutic inhalation of two essential oils ameliorates scopolamine-induced cognitive impairment in mice.

Clover and lemongrass essential oils of contrasting composition, at three concentration levels (1%, 5%, 10%), were administrated via prophylactic and therapeutic inhalation to scopolamine-treated mice. Chemical analysis showed that clover oil was dominant in eugenol (47.69%) and lemongrass free of eugenol but mainly containing monoterpenoids of comparable proportions. Animal behavioural and brain biochemical tests showed that injection of scopolamine caused memory and learning deficit in mice while prophylactic and therapeutic inhalation of two oils at moderate to high concentrations all obviously reversed the cognitive impairment via inhibiting acetylcholinesterase activities, oxidation and inflammation. Lemongrass essential oil with diverse monoterpenoids can be as effective as or a little bit more potent than eugenol-rich clover essential oil possibly due to the synergistic effect of various monoterpenoids. These findings implied that sniffing of such aroma recipes could be a promising complementary approach for the mitigation of Alzheimer's disease-related cognitive impairment.

Evaluation of compressive strength, microhardness and solubility of zinc-oxide eugenol cement reinforced with E-glass fibers.

BACKGROUND: Zinc-oxide eugenol (ZOE) cements are among the most used temporary materials in dentistry. Although ZOE has advantages over other temporary fillers, its mechanical strength is weaker, so researchers are working to improve it. E-glass fibers have emerged as promising reinforcing fibers in recent years due to their strong mechanical behavior, adequate bonding, and acceptable aesthetics. OBJECTIVES: To evaluate and compare the compressive strength, surface microhardness, and solubility of the ZOE and those reinforced with 10 wt.% E-glass fibers. METHODS: A total of 60 ZEO specimens were prepared; 30 specimens were reinforced with 10 wt.% E-glass fibers, considered modified ZOE. The characterization of the E-glass fibers was performed by XRF, SEM, and PSD. The compressive strength, surface microhardness, and solubility were evaluated. Independent sample t-tests were used to statistically assess the data and compare mean values (P ≤ 0.05). RESULTS: The results revealed that the modified ZOE showed a significantly higher mean value of compressive strength and surface microhardness while having a significantly lower mean value of solubility compared to unmodified ZOE (P ≤ 0.05). CONCLUSION: The modified ZOE with 10 wt.% E-glass fibers had the opportunity to be used as permanent filling materials.

Paeonia mascula subsp. russoi (biv.) Cullen & Heywood: the chemical composition of the aerial parts essential oils of two different populations collected in Sicily (Italy).

In the present study, the chemical composition of the essential oil from aerial parts of two populations of Paeonia mascula subsp. russoi, collected in Sicily, was evaluated by GC-MS. No previously phytochemical investigation has been reported for this subspecies. The main components of the essential oil of the population with pink flowers were salicylaldehyde (34.31%), nonanal (16.95%) and 2-hexenal (10.17%), whereas essential oil of the population with white flowers, was shown to be rich of myrtanal (14.14%), eugenol (14.02%) and salicylaldehyde (12.21%). Furthermore, a complete literature review, not present in literature, on the composition of the essential oils of all the other taxa of Paeonia, studied so far, was performed. PCA and HCA analyses of the composition of essential oils obtained from the aerial parts were also carried out.

On the antimicrobial properties and endurance of eugenol and 2-phenylphenol functionalized sol-gel coatings.

Preventing microbiological surface contamination in public spaces is nowadays of high priority. The proliferation of a microbial infection may arise through air, water, or direct contact with infected surfaces. Chemical sanitization is one of the most effective approaches to avoid the proliferation of microorganisms. However, extended contact with chemicals for cleaning purposes such as chlorine, hydrogen peroxide or ethanol may lead to long-term diseases as well as drowsiness or respiratory issues, not to mention environmental issues associated to their use. As a potentially safer alternative, in the present work, the efficacy and endurance of the antimicrobial activity of different sol-gel coatings were studied, where one or two biocides were added to the coating matrix resulting on active groups exposed on the surface. Specifically, the coating formulations were synthesized by the sol-gel method. Using the alkoxide route with acid catalysis a hybrid silica-titania-methacrylate matrix was obtained where aromatic liquid eugenol was added with a double function: as a complexing agent for the chelation of the reaction precursor titanium isopropoxide, and as a biocide. In addition, 2-Phenylphenol, ECHA approved biocide, has also been incorporated to the coating matrix. The antibacterial effect of these coatings was confirmed on Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (Escherichia coli). Additionally, the coatings were non cyto-toxic and displayed virucidal activity. The coating chemical composition was characterized by 29Si NMR, and ATR-FTIR. Furthermore, the thickness and the mechanical properties were characterized by profilometry and nanoindentation, respectively. Finally, the durability of the coatings was studied with tribology tests. Overall, our data support the efficacy of the tested sol-gel coatings and suggest that added features may be required to improve endurance of the antimicrobial effects on operational conditions.

Role of marination, natural antimicrobial compounds, and packaging on microbiota during storage of chicken tawook.

The aim of this study was to investigate the antimicrobial effect of marination, natural antimicrobials, and packaging on the microbial population of chicken tawook during storage at 4°C. Chicken meat was cut into 10 g cubes and marinated. The chicken was then mixed individually with 0.5% or 1% (w/v) vanillin (VA), ß-resorcylic acid (BR), or eugenol (EU), and stored under aerobic (AP) or vacuum (VP) packing at 4°C for 7 d. The marinade decreased microbial growth as monitored by total plate count, yeast and mold, lactic acid bacteria, and Pseudomonas spp. by about 1 log cfu/g under AP. The combination of marinade and antimicrobials under AP and VP decreased growth of spoilage-causing microorganisms by 1.5 to 4.8 and 2.3 to 4.6 log cfu/g, respectively. Change in pH in VP meat was less than 0.5 in all treated samples including the control. Marination decreased the lightness of the meat (L*) and significantly (p < 0.05) increased the redness (A*) and yellowness (B*). Overall acceptability was highest for marinated samples with 0.5% BR.

Cardioprotective Effect of Eugenol Against Cd-Induced Inflammation, Oxidative Stress, and Dyslipidemia in Male Rats: An In Vivo and Molecular Docking Study.

Cadmium, a highly toxic heavy metal, can cause severe damage to several vital organs including the kidney, liver, and brain. Many of the natural compounds found in aromatic plants have beneficial pharmacological properties. Eugenol is one such compound reported to have anti-inflammatory and antioxidant properties. The aim of this study is to investigate whether eugenol, a natural compound found in aromatic plants known for its anti-inflammatory and antioxidant properties, can mitigate the detrimental effects of cadmium exposure on cardiac inflammation, oxidative stress, and dyslipidemia. Male albino rats were subjected to randomization into four groups, each comprising six animals, to investigate the potential of eugenol in mitigating cadmium-induced toxicity. All groups received oral gavage treatment for 21 days. Following the treatment regimen, cardiac tissue specimens were collected for analysis. The assessment of cardiac antioxidant status entailed the determination of enzymatic activities including catalase, SOD, GST, and GPx. Additionally, levels of lipid peroxidation, reduced glutathione, protein carbonyl oxidation, and thiol levels were quantified in the cardiac tissue samples. To evaluate cardiac damage, marker enzymes such as LDH and CK-MB were measured. Furthermore, the inflammatory response in the cardiac tissue induced by cadmium exposure was assessed through the quantification of NO, TNF-α, and IL-6 levels. Additionally, molecular docking and dynamics studies were conducted utilizing autodock and GLIDE methodologies. Cadmium administration markedly enhanced the activities of LDH and CK-MB, prominent cardiac markers. Furthermore, cadmium treatment also demonstrated a significant decrease in the reduced glutathione levels and antioxidant enzyme activities. Significant elevation of the inflammatory markers was also observed in the cadmium-treated group. Eugenol treatment effectively ameliorates cadmium-induced biochemical changes. This study underscores the potent anti-inflammatory and antioxidant attributes of eugenol. Co-administration of eugenol alongside cadmium exhibited remarkable protective efficacy against cadmium-induced cardio-toxicity. Eugenol demonstrated the capability to reinstate the cellular redox equilibrium of rats subjected to cadmium treatment to levels akin to those of the normal control group.

Development of broad-spectrum immunoassay with monoclonal antibody to detect five eugenols and study of their molecular recognition mechanism.

In this study, three eugenol fragment-containing haptens were synthesized, and a monoclonal antibody (mAb) selective for five commonly-found eugenol compounds (EUGs, i.e., eugenol, isoeugenol, methyl eugenol, methyl isoeugenol, and acetyl isoeugenol) was obtained. Based on this mAb, a broad-spectrum indirect competitive ELISA for high-throughput detection of five EUGs was developed. The detection limits for eugenol, isoeugenol, methyl eugenol, methyl isoeugenol and acetyl isoeugenol in both tilapia and shrimp samples were 25.3/ 50.6 µg/kg, 0.075/0.15 µg/kg, 0.48/0.96 µg/kg, 0.16/0.32 µg/kg, and 18.16/36.32 µg/kg, respectively. The recoveries for five EUGs ranged from 80.4 to 114.0 % with a coefficient of variation less than 11.5 %. Moreover, homology modelling and molecular docking were conducted to elucidate the interactions mechanism of mAb-EUGs. The work provides a promising tool for high-throughput screening of EUGs in aquatic products, which can serve as a benchmark for designing haptens and developing immunoassays for other small molecules.

Protective role of Eugenol against the destructive effects of lead on conditioned fear memory in male rats with post-traumatic stress disorder-related behavioral traits.

Introduction: Post-traumatic stress disorder (PTSD) is a consequence of living in today's stressful society. Patients have difficulty forgetting traumatic events. lead pollution has many effects on the nervous system, one of which is memory and learning disorders. The herbal medicine Eugenol has a beneficial effect on memory. Aim: This study aims to investigate the protective effect of Eugenol on lead-induced memory impairments in stressed rats. Methods: In the first experiment, the animals were divided into three groups: SPS+Saline, SPS+Pb, and naïve. The SPS+Saline, SPS+Pb groups received normal saline and lead through gavage for 21 days, while the sham group remained untreated. Rats were subjected to the modified single prolonged stress model. Memory tests were conducted one week later, evaluating freezing levels in three consecutive tests over three days. In the second experiment, rats were divided into a SPS+Pb+Saline and three treatment groups. The SPS+Pb+Saline group received daily saline injections, while the other groups received different doses of Eugenol (25, 50, and 100 mg/kg). Memory tests similar to the first experiment were conducted. Results: The results showed significantly higher immobility levels in the SPS+Saline and SPS+Pb groups compared to the sham. Additionally, the SPS+Pb group had a significant higher immobility compared to the SPS+Saline group. In the second experiment, the SPS+Pb+EU 25 group showed a significant lower freezing compared to the SPS+Pb+Saline group. Additionally, freezing in the SPS+Pb+EU 50 and SPS+Pb+EU 100 groups was significantly higher than in the SPS+Pb+EU 25 group. The SPS+Pb+EU 50 group showed a significant higher freezing compared to the SPS+Pb+Saline group. Conclusion: lead acetate exacerbated memory impairments in stressed rats and Eugenol, particularly at a dose of 25 mg/kg, improved these impairments. Therefore, Eugenol has the potential to partially reduce the negative effects of lead on memory in individuals with PTSD.

Molecular Docking Analysis of Hydroxyclavicol and Eugenol From Betel Leaves Against Outer Membrane Protein (OmpH) of Dialister pneumosintes.

Introduction Dialister pneumosintes is an obligate anaerobic non-spore-forming Gram-negative bacilli. As a part of polymicrobial film, the activated virulence factor causes oral diseases like gingivitis and periodontitis. Decreased susceptibility of clinical strains of D. pneumosintes to different antibiotics including piperacillin and metronidazole raises concerns. There has been significant interest in the utility of plant phytocompounds as potent antibacterial agents.  Aim The study aimed to look at the potential of two phytocompounds, eugenol and hydroxychavicol, for their ability to inhibit outer membrane protein (OmpH) of D. pneumosintes using computational tools. Results The study showed effective inhibition of the OmpH of D. pneumosintes by both eugenol and hydroxychavicol. The high probability to be active (Pa) value indicated the probability of true positive for the tested compounds for their predicted biological activity. There was strong reciprocity between the drug-likeliness and its binding affinity for the target protein, indicating an inhibitory nature. Conclusion The tested phytocompounds hydroxychavicol and eugenol showed potential inhibition of the OmpH protein of D. pneumosintes indicating its potential use as inhibitory compounds of the pathogen and future directions for the treatment of periodontitis and gingivitis.

Morphometric parameters of dental pulp in immature teeth in a sheep model after mechanical pulp exposure and restoration with reinforced zinc oxide-eugenol.

Background: The aim of the study was to investigate the morphometric parameters of dental pulp in open apices immature teeth in a sheep model after mechanical pulp exposure and restoration with reinforced zinc oxide-eugenol. Materials and Methods: In this experimental study, a total of 12 immature mandibular central incisors from six adult male sheep, weighing 30-40 kg and with the age of 1 year old with Merino race were examined. After anesthesia, the pulps of the teeth in the case group were mechanically exposed and then were restored with reinforced zinc oxide-eugenol and amalgam. In the control group, the teeth remained intact. The animals were sacrificed at intervals of 2, 4, 6, and 8 weeks (E2, E4, E6, and E8) in the case and 2 and 8 weeks (C2 and C8) in the control groups. Then, their teeth were removed with the surrounding supporting tissues and alveolar bones. Tissue processing and staining were done, and the sections were examined under a light microscope. The Kruskal-Wallis and Mann-Whitney U tests were used to analyze the data and compare the changes between the two groups. P < 0.05 was considered statistically significant. Results: In response to mechanical exposure, reparative or tertiary dentin was formed, and its thickness increased during the time of the study. The thickness of the odontoblastic layer in the E4 group was the highest amount. The pulp chamber diameter in the C2 group was significantly larger than the other groups, and the diameter of the apical foramen in the E8 was decreased significantly compared to the controls (P < 0.05). Conclusion: In response to mechanical exposure and restoration with reinforced zinc oxide-eugenol, some morphometric parameters of the dental pulp changed significantly in the sheep model compared to the controls.

Antimicrobial Activity of Syzygium aromaticum Essential Oil in Human Health Treatment.

The use of natural compounds to prevent and treat infective diseases is increasing its importance, especially in the case of multidrug-resistant (MDR) microorganisms-mediated infections. The drug resistance phenomenon is today a global problem, so it is important to have available substances able to counteract MDR infections. Syzygium aromaticum (L.) Merr. & L.M. Perry (commonly called clove) is a spice characterized by several biological properties. Clove essential oil (EO) consists of numerous active molecules, being eugenol as the principal component; however, other compounds that synergize with each other are responsible for the biological properties of the EO. S. aromaticum is traditionally used for bowel and stomach disorders, cold and flu, oral hygiene, tooth decay, and for its analgesic action. Its EO has shown antioxidant, antimicrobial, anti-inflammatory, neuro-protective, anti-stress, anticancer, and anti-nociceptive activities. This review aims to investigate the role of E. S. aromaticum EO in the counteraction of MDR microorganisms responsible for human disorders, diseases, or infections, such as Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhi, Candida albicans, Giardia lamblia, Streptococcus mutans, Porphyromonas gingivalis, and Klebsiella pneumoniae. This study might orient clinical researchers on future therapeutic uses of S. aromaticum EO in the prevention and treatment of infectious diseases.

Eugenol Possesses Colitis Protective Effects: Impacts on the TLR4/MyD88/NF-[Formula: see text]B Pathway, Intestinal Epithelial Barrier, and Macrophage Polarization.

Eugenol (EU) has been shown to ameliorate experimental colitis due to its anti-oxidant and anti-inflammatory bioactivities. In this study, DSS-induced acute colitis was established and applied to clarify the regulation efficacy of EU on intestinal barrier impairment and macrophage polarization imbalance along with the inflammatory response. Besides, the adjusting effect of EU on macrophages was further investigated in vitro. The results confirmed that EU intervention alleviated DSS-induced colitis through methods such as restraining weight loss and colonic shortening and decreasing DAI scores. Microscopic observation manifested that EU maintained the intestinal barrier integrity in line with the mucus barrier and tight junction protection. Furthermore, EU intervention significantly suppressed the activation of TLR4/MyD88/NF-[Formula: see text]B signaling pathways and pro-inflammatory cytokines gene expressions, while enhancing the expressions of anti-inflammatory cytokines. Simultaneously, WB and FCM analyses of the CD86 and CD206 showed that EU could regulate the DSS-induced macrophage polarization imbalance. Overall, our data further elucidated the mechanism of EU's defensive effect on experimental colitis, which is relevant to the protective efficacy of intestinal barriers, inhibition of oxidative stress and excessive inflammatory response, and reprogramming of macrophage polarization. Hence, this study may facilitate a better understanding of the protective action of the EU against UC.

Effects of Aspirin Eugenol Ester on Liver Oxidative Damage and Energy Metabolism in Immune-Stressed Broilers.

The aim of this study was to investigate the effects of aspirin eugenol ester (AEE) on liver oxidative damage and energy metabolism in immune-stressed broilers. In total, 312 broilers were divided into 4 groups (saline, LPS, SAEE, and LAEE). Broilers in the saline and LPS groups were fed a basal diet; the SAEE and LAEE groups had an added 0.01% AEE in their diet. Broilers in the LPS and LAEE groups were injected with lipopolysaccharides, while the saline and SAEE groups were injected with saline. Results showed that AEE increased the body weight, average daily gain, and average daily feed intake, as well as decreasing the feed conversion ratio of immune-stressed broilers. AEE protects against oxidative damage in immune-stressed broiler livers by elevating the total antioxidant capacity, superoxide dismutase activity, and glutathione S-transferase alpha 3 (GSTA3) and glutaredoxin 2 (GLRX2) expression, while decreasing malondialdehyde content. AEE lessened inflammation by reducing prostaglandin-F2α production and prostaglandin-endoperoxide synthase 2 (PTGS2) and interleukin-1beta (IL-1ß) expression. AEE decreased oxidative phosphorylation rates by increasing succinic acid levels and lowering both adenosine diphosphate (ADP) levels and ceroid lipofuscinosis neuronal 5 (CLN5) expression. AEE modulated the metabolism of phenylalanine, tyrosine, lipids, and cholesterol by reducing the phenyllactate and L-arogenate levels, lowering dopachrome tautomerase (DCT) and apolipoprotein A4 (APOA4) expression, and increasing phenylpyruvic acid and dopa decarboxylase (DDC) expression. In summary, AEE can effectively alleviate liver oxidative damage and energy metabolism disorders in immune-stressed broilers.

In silico and in vivo evaluation of the anti-cryptosporidial activity of eugenol.

Background: Cryptosporidiosis is an opportunistic parasitic disease widely distributed worldwide. Although Cryptosporidium sp. causes asymptomatic infection in healthy people, it may lead to severe illness in immunocompromised individuals. Limited effective therapeutic alternatives are available against cryptosporidiosis in this category of patients. So, there is an urgent need for therapeutic alternatives for cryptosporidiosis. Recently, the potential uses of Eugenol (EUG) have been considered a promising novel treatment for bacterial and parasitic infections. Consequently, it is suggested to investigate the effect of EUG as an option for the treatment of cryptosporidiosis. Materials and methods: The in silico bioinformatics analysis was used to predict and determine the binding affinities and intermolecular interactions of EUG and Nitazoxanide (NTZ) toward several Cryptosporidium parvum (C. parvum) lowa II target proteins. For animal study, five groups of immunosuppressed Swiss albino mice (10 mice each) were used. Group I was left uninfected (control), and four groups were infected with 1,000 oocysts of Cryptosporidium sp. The first infected group was left untreated. The remaining three infected groups received NTZ, EUG, and EUG + NTZ, respectively, on the 6th day post-infection (dpi). All mice were sacrificed 30 dpi. The efficacy of the used formulas was assessed by counting the number of C. parvum oocysts excreted in stool of infected mice, histopathological examination of the ileum and liver tissues and determination of the expression of iNOS in the ileum of mice in different animal groups. Results: treatment with EUG resulted in a significant reduction in the number of oocysts secreted in stool when compared to infected untreated mice. In addition, oocyst excretion was significantly reduced in mice received a combination therapy of EUG and NTZ when compared with those received NTZ alone. EUG succeeded in reverting the histopathological alterations induced by Cryptosporidium infection either alone or in combination with NTZ. Moreover, mice received EUG showed marked reduction of the expression of iNOS in ileal tissues. Conclusion: Based on the results, the present study signified a basis for utilizing EUG as an affordable, safe, and alternative therapy combined with NTZ in the management of cryptosporidiosis.