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Chicken meat and its derivatives are easily alterable. They are a nutritionally healthy food, and their consumption has seen a remarkable increase worldwide in recent years. At the same time, consumer demand for the use of natural products to control microbial growth is increasing. In this context, the antimicrobial capacity of a commercial extract of the lemon verbena (Lippia citriodora) plant, (LCE) was tested in binary combination with gallic acid or octyl gallate against two strains of lactic acid bacteria (LAB) of meat origin: Carnobacterium divergens ATCC 35677 and Leuconostoc carnosum ATCC 49367. First, the antimicrobial potential was evaluated by the checkerboard microdilution method at the optimal growth temperature of each and at 4 °C, pH 5.7 and 6.7, in culture medium. Octyl gallate was the most effective antimicrobial against the two bacteria under all study conditions. At 4 °C, the combination of LCE with octyl gallate had a similar antimicrobial effect on the two LAB, being bactericidal at pH 6.7. In chicken breast, this effective combination was tested in normal or modified atmosphere and refrigerated (4-8 °C) for 9 days. LCE + OG in modified atmosphere reduced the different microbial groups studied, including the lactic acid bacteria as the main microorganisms responsible for the spoilage of fresh meat. Further research could pave the way for the development of novel strategies contributing to the technological stability, security, and functional properties of chicken meat.

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Lemon verbena infusions are widely appreciated due to their agreeable lemony flavor and medicinal properties. In this study, the antioxidant potential, phenolic profile, and free amino acid profile of lemon verbena infusions from different commercial brands were studied. Characterization by UHPLC-QTOF-HRMS allowed the identification of 34 phenolics. The free amino acid profile (by RP-HPLC-FLD) was assessed for the first time, allowing the quantification of 16 amino acids. Furthermore, the infusions showed high antioxidant activity by different assays (ferric reducing antioxidant power, DPPH• scavenging, and oxygen radical absorbance capacity assays), which in turn were significantly correlated with total phenolics and total flavonoid contents. Notwithstanding, phenylalanine seemed to have also an impact on the antioxidant activity of the infusions, with significant correlations found. Finally, significant differences were found in all the evaluated parameters for one of the four commercial brands herein studied, which was possibly related to the different geographical origins of this sample. Overall, these lemon verbena infusions proved to be rich in a huge variety of bioactive compounds that can provide therapeutic potential.

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Objective: Acrylamide (ACR) neurotoxicity is induced by different mechanisms such as oxidative stress and apoptosis. Scientific researchs have indicated the antioxidative properties of Lippia citriodora. The protective effect of L. citriodora aqueous and ethanolic extracts on ACR-induced neurotoxicity was investigated. Materials and methods: Male Wistar rats were randomly divided into 13 groups: (1) control, (2) ACR (50 mg/kg, i.p.), (3-6) ACR+aqueous extract (12.5, 25, 50, and 100 mg/kg, i.p.), (7-10) ACR+ethanolic extract (12.5, 25, 50, and 100 mg/kg, i.p.), (11) aqueous extract (100 mg/kg), (12) ethanolic extract (100 mg/kg), and (13) ACR+Vitamin E (200 mg/kg, every other day, i.p.). After 11 days, gait score, MDA, and GSH levels in brain cortical tissue were measured. In the in vitro test, the viability of PC12 cells (using MTT test), the amount of reactive oxygen species (ROS; using DCFH-DA method), and the protein levels of Bax, Bcl2 and caspase 3 (by western blotting) were measured. Results: In the in vitro study, the IC50 for the treatment of PC 12 cells with ACR after 24 hr was 6 mM. ACR decreased cell viability, but increased ROS level, Bax/Bcl-2 ratio, and caspase-3 protein level. Pre-treatment by L. citriodora extracts (15-120 µg/ml) ameliorated the toxic effects of ACR on PC12 cells. In the in vivo experiment, ACR-induced movement disorders increased MDA but decreased GSH content. The extracts of L. citriodora improved ACR toxic effects. Conclusion: Aqueous and ethanolic extracts of L. citriodora were found to reduce ACR-induced neurotoxicity via inhibiting oxidative stress and apoptosis.

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ABSTRACT: The antimicrobial activity of a commercial Lippia citriodora extract (LCE) at 2,500 µg mL-1 (maximum sensory acceptable level) and vanillin (MIC and 2 MIC) alone and in combination were analyzed in four strains of Escherichia coli (two nonverotoxigenic and two verotoxigenic) in cation-adjusted Mueller-Hinton medium and in Piel de Sapo melon juice (MJ) stored under refrigeration (4°C) for 7 days. The bacterial counts of the four strains together in untreated samples were higher (≈4 log CFU/mL) in cation-adjusted Mueller-Hinton medium than in stored MJ. LCE showed higher antimicrobial activity in MJ than in standard culture broth, but vanillin showed a higher effect in broth. The verotoxigenic strain E. coli O146:H stx2 was the most sensitive to LCE in refrigerated MJ. Combinations of vanillin (at MIC and 2 MIC) with LCE were very effective in reducing E. coli counts either in broth or in refrigerated MJ to undetectable levels. Bactericidal effects were observed for the combinations in all strains in broth and in MJ. Also, these combinations showed an antimicrobial synergistic effect after day 3 of storage in MJ in three of the bacterial strains tested. These results indicate that the combination of LCE (at maximum sensory acceptable levels) and vanillin (at low concentrations) could be considered as a promising natural antimicrobial agent to inhibit verotoxigenic E. coli growth in refrigerated MJ and improve its quality.

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INTRODUCTION: Polyphenols have shown capacity to improve appetite sensation, leading to enhanced control of body weight. However, despite being related with hunger-related hormones, metabolic and mechanism are not well known. METHODS: The effectiveness of a nutraceutical composed of extract to Lippia citriodora and Hibiscus sabdarrifa (Lc-Hs) for controlling satiety and hunger was analyzed in a cross-over, placebo-controlled (Pla) clinical intervention. The study was divided in two 60-day periods separated by 30-day length wash-out period. At the end of each period, overweight and obese subjects (n = 33; age = 33.76 ± 12.23; BMI = 28.20 kg/m2 ± 2.47; fat mass 30.65 ± 8.39%; both sexes were proposed to eat an ad-libitum meal. Meanwhile, appetite sensation was determined by visual analog scales at different times. Moreover, blood extraction was performed to determine biochemical parameters (lipid and glucidic profile and safety parameters) and to evaluate hunger-related hormones (insulin, leptin, ghrelin, adiponectin, GLP-1 and peptide YY). RESULTS: A decrease in appetite sensation was observed in Lc-Hs treatment, showing higher satiety quotient (Pla = 3.36 ± 2.33%mm/kcal; Lc-Hs = 5.53 ± 2.91%mm/kcal; p < 0.0001). Area under the curve was higher in Pla compared to Lc-Hs during the test, from baseline to minute 240 (240 (Pla 9136.65 ± 2261.46% x min-1; Lc-Hs 8279.73 ± 2745.71% x min-1; p < 0.014). Energy consumption was lower for subjects treated with Lc-Hs (774.44 ± 247.77 kcal) compared to those treated with Pla (849.52 ± 246.54 kcal) (p < 0.004). Leptin values varied from baseline (Pla 12.36 ± 1.98 ng/mL; Lc-Hs 13.13 ± 1.99 ng/mL) to the end of the study (Pla 12.60 ± 2.02 ng/mL; Lc-Hs 12.06 ± 2.05 ng/mL; p < 0.047). GLP-1 values varied (p < 0.001) in Lc-Hs treatment from baseline (4.34 ± 0.49 ng/mL) to the end of the study (3.23 ± 0.52 ng/mL). CONCLUSION: The supplementation with the Lc-Hs extract decreases appetite sensation in overweight and obese population, reducing calorie intake after an ad-libitum meal. Due to variation on hunger-related hormones and the relationship between satiety feeling, it would be interesting to develop future research focused on the variation of the hormones themselves.

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OBJECTIVE: Coronary artery disease (CAD) is among the most prevalent diseases in the world, and its severity is usually assessed through coronary angiography (CA). Anxiety is the most prevalent problem before angiography. The aim of this study was to evaluate Crocus sativus (saffron) and Lippia citriodora Kunth (lippia) extracts and saffron-lippia extract combination on anxiety among the candidates for CA. MATERIALS AND METHODS: This double-blind randomized placebo-controlled trial in four month was conducted on 120 angiography candidates in Valiasr hospital, Birjand, Iran. The random allocation was doing through block randomization to saffron, lippia, saffron-lippia, and placebo groups; four hours before angiography, participants in these groups respectively received a single dose of saffron extract capsule (40 mg), lippia extract capsule (40 mg), saffron-lippia extract capsule (20 mg saffron and 20 mg lippia), and lactulose capsule (40 mg). Participants' anxiety was assessed before, thirty minutes after, and three hours after the intervention via Spielberger State-Trait Anxiety Inventory. RESULTS: The mean scores of state, trait, and total anxiety in the saffron group reduced significantly (p<0.05), while the mean scores in the other groups did not change significantly, except for the mean score of trait anxiety in the saffron-lippia group which decreased significantly (p=0.05). Therefore, after the intervention, the mean scores of state, trait, and total anxiety in the saffron group were significantly lower than the other groups (all, p<0.05). CONCLUSION: This study suggests that the oral use of a single-dose of 40 mg saffron extract is effective in alleviating anxiety among the candidates for CA.

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Lippia citriodora is a flowering plant cultivated for its lemon-scented leaves and used in folk medicine for the preparation of tea for the alleviation of symptoms of gastrointestinal disorders, cold, and asthma. The oil extracted from the plant leaves was shown to possess antioxidant potential and to exert antiproliferative activity against breast cancer. The aim of this study was to further investigate potential antitumor effects of L. citriodora oil (LCO) on breast cancer. The in vitro antiproliferative activity of LCO was examined against murine DA3 breast cancer cells by the sulforhodamine B assay. We further explored the LCO's pro-apoptotic potential with the Annexin-PI method. The LCO's anti-migratory effect was assessed by the wound-healing assay. LCO was found to inhibit the growth of DA3 cells in vitro, attenuate their migration, and induce apoptosis. Finally, oral administration of LCO for 14 days in mice inhibited by 55% the size of developing tumors in the DA3 murine tumor model. Noteworthy, in the tumor tissue of LCO-treated mice the apoptotic marker cleaved caspase-3 was elevated, while a reduced protein expression of survivin was observed. These results indicate that LCO, as a source of bioactive compounds, has a very interesting nutraceutical potential.

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Hypertension is an important factor of cardiovascular diseases and contributes to their negative consequences including mortality. The World Health Organization estimated that 54% of strokes and 47% of cases of ischemic heart illness are related to high blood pressure. Recently, Hibiscus sabdariffa (HS) and Lippia citriodora (LC) have attracted scientific interest, and they are recognized for their high content of polyphenols as these may prevent several disease factors, such as hypertension. The aim of the present study is to determine if supplementation with an HS-LC blend (MetabolAid®) may be effective for the treatment of type 1 hypertensive sedentary populations. A total of 80 type 1 hypertensive subjects of both sexes were included in the study and were treated with placebo or the HS-LC extract, and both groups were treated over 84 days. The blood pressure (diastolic, systolic, and pulse pressure) was measured throughout the day, for each of the days of the study duration and determined using Ambulatory Blood Pressure Monitoring (ABPM). Physical activity was determined throughout the study to ensure similar conditions related to exercise. The results showed the capacity for reducing the blood pressure parameters in the case of the HS-LC extract. The daily consumption of the HS-LC extract but not the placebo over 84 days was able to reduce the daytime parameters related to blood pressure. The most remarkable results were observed in the measurements performed during the daytime, especially in the systolic blood pressure showing statistically significant variation.

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Pressurized liquid extraction was performed to obtain a phytocomplex from Lippia citriodora leaves rich in bioactive compounds. The extract was loaded in phospholipid vesicles to improve its protective effect against oxidative stress in the intestine. The phytochemicals were identified and quantified by HPLC-ESI-TOF-MS. The extract was incorporated in liposomes and penetration enhancer-containing vesicles (PEVs) modified with glucidex, a dextrin, and a biopolymer obtained from Chimaera monstrosa. The PEVs were smaller than liposomes (~150 vs 370 nm) and more stable, according to accelerated aging tests. The integrity of the vesicles in acidic or neutral pH and high ionic strength or in milk whey was assessed. The cytocompatibility of the formulations and their ability to protect Caco-2 cells against oxidative stress were confirmed in vitro and compared with two commercial extracts of L. citriodora. The results confirmed the suitability of formulations to be used in functional foods to protect the intestine from oxidative stress.

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AIM: The study was conducted to assess the antidiabetic and antihyperlipidemic effects of Lippia citriodora. BACKGROUND: Lippia citriodora Kunth (Verbenaceae) is a medicinal plant with many traditional uses. OBJECTIVE: The goal of this study was to investigate the antidiabetic activity of aqueous extract of Lippia citriodora and its capacity to improve the lipid profile in diabetic rats. METHODS: Normal and diabetic rats were treated orally for 15 days by the leaves aqueous extract of Lippia citriodora (LCLAE) (100 mg/kg). The whole glucose utilization and liver architecture were also evaluated. The in vitro antioxidant activity of LCLAE was also assessed. RESULTS: LCLAE lowered blood glucose and lipid levels in all groups. Moreover, LCLAE ameliorated the liver structure and exhibited antioxidant activity. CONCLUSION: In conclusion, this study demonstrates the hypoglycemic and antihyperlipidemic effects of LCLAE in rats.

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Phenolic compounds have shown to have a high bioactive potential against various pathologies, postulating as an interesting alternative to manage some diseases. In this sense, both Lippia citriodora and Hibiscus sabdariffa are two botanical sources with a demonstrated high bioactive potential, in which their antioxidant capacity stands out. In this work, the optimization of the extraction conditions for the recovery of phytochemicals from L. citriodora leaves and H. sabdariffa calyces has been carried out using Response Surface Methodologies (RSM) considering their total polar compounds measured by HPLC-ESI-TOF/MS and Folin-Ciocalteu assay, and its antioxidant capacity evaluated by Ferric Reducing Antioxidant Power (FRAP) and Trolox Equivalent Antioxidant Capacity (TEAC) assays. The results showed that to maximize the antioxidant capacity in H. sabdariffa, a moderate temperature and high ethanol percentage are needed, while a low temperature and a high percentage of ethanol are needed in L. citriodora. In addition, with the results obtained in the multiple response analysis, it is possible to affirm the importance of this type of analysis to develop functional ingredients, taking into account both total content of phenolic compounds and their bioactivity. Furthermore, as confirmed in this study, these analyses can be extrapolated in different techniques and in different matrices, with phenolic compounds from different families being important to develop new high added value products for food, pharmaceutical or cosmetic industries.

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Lippia citriodora has been demonstrated to have a wide variety of phytochemicals which provide benefits to human health acting as antioxidants or anti-obesogenics. In this study, these phytochemicals were recovered using a microwave-assisted technology and applying optimal conditions and microencapsulated using spray drying. In this study, two different carbohydrates, maltodextrin (MD) and inulin (IN), were compared as carriers in the encapsulation procedure. The spray drying process was optimized by using a response surface methodology (RSM) based on a central composite design 22, where air inlet temperature and the sample:encapsulating agent ratio (S:EA) were selected as independent variables. Both designs were analyzed equally to evaluate differences between each carrying agent on polar compounds' encapsulation (process yield (Y%), encapsulation efficiency (EE%) and recovery of compounds (R%)) during the spray drying. The EE% and R% of each polar compound was monitored by High Performance Liquid Chromatography coupled to Time-of-Flight mass spectrometer by electrospray interface (HPLC-ESI-TOF-MS). The results showed that the use of IN as a carrier increased the powder recovered and the recovery of polar compounds after the spray dry process, whereas MD achieved a higher encapsulation efficiency.

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The aim of this work was to optimize the recovery of phenolic compounds from Lippia citriodora using supercritical fluid extraction (SFE). To achieve this goal, response surface methodology based on a 23 central composite design was used to evaluate the effects of the following experimental factors: temperature, pressure and co-solvent percentage. The effects of these variables on the extraction yield and total polar compound contents were evaluated. With respect to the phytochemical composition, an exhaustive individual phenolic compound quantitation was carried out by HPLC-ESI-TOF-MS to analyze the functional ingredients produced by this system design. To the best of our knowledge, this is the first time that a standardized supercritical fluid process has been developed to obtain functional ingredients based on phenolic compounds from L. citriodora in which the individual compound concentration was monitored over the different SFE conditions. The results enabled the establishment of the optimal technical parameters for developing functional ingredients and revealed the main factors that should be included in the extraction process control. This functional food ingredient design could be used as a control system to be applied in nutraceutical and functional food production industry.

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SCOPE: Obesity is characterized by a dysfunction in the adipose tissue and an inflammatory subclinical state leading to insulin resistance and increased risk of cardiovascular diseases. It is also associated with intestinal dysbiosis that contributes to inflammation development. Lippia citriodora (LCE) contains high levels of polyphenolpropanoids and has shown promising results in obesity. The aim of this study is to investigate a well-characterized extract of LCE in a model of metabolic syndrome in mice, focusing on its effects on metabolic tissues, endothelial dysfunction, and microbiome. METHODS: Mice are fed a high fat diet (HFD) for six weeks and treated daily with LCE (1, 10, and 25 mg kg-1 ). Glucose and lipid metabolism is investigated. The inflammatory state in the metabolic tissues and the intestinal microbiota composition are characterized, as well as the endothelium-dependent vasodilator response to acetylcholine. RESULTS: LCE reduces fat accumulation and improves plasma glycemic and lipid profiles, as well as the inflammatory process and vascular dysfunction. Moreover, LCE lessens intestinal dysbiosis, as it reduces the Firmicutes/Bacteroidetes ratio and increases Akkermansia abundance in comparison with untreated HFD mice. CONCLUSION: The antiobesity therapeutic properties of LCE are most probably mediated by the synergic effects of its bioactive compounds.

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Phytochemicals from Lippia citriodora leaves were extracted by applying an innovative technology based on the use of microwaves, which represents an alternative method to extract bioactive substances. The obtained extract was incorporated into phospholipid vesicles in order to promote the antioxidant effect of the bioactive molecules present in L. citriodora extract. The extract was analyzed by High Performance Liquid Chromatography coupled to Time-Of-Flight mass spectrometer by electrospray (HPLC-ESI-TOF-MS) and different phytochemicals were detected and quantified. The whole extract was incorporated in liposomes, glycerosomes (liposomes modified with glycerol) and propylene glycol-containing vesicles (PG-PEVs). Moreover, a biopolymer obtained from fish by-product, that is Thunnus albacares skin, was added to improve the bioactivity of the formulations. The in vitro biocompatibility and the antioxidant efficacy of the extract in solution or loaded in the vesicles were tested in primary mouse embryonic fibroblasts (3T3). The results showed the superior bioactivity of the vesicle formulations over the aqueous solution of the extract, which points to an interesting strategy for the treatment of skin disorders.

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: The location and quantity of body fat determine the health risks, limiting people with obesity. Recently, polyphenols have attracted the attention of the scientific community because of their potential use for the reduction of obesity. A proprietary formula comprised of a blend of Lippia citriodora and Hibiscus sabdariffa has been recognized for its high content of polyphenols, powerful antioxidant molecules that may prevent weight gain and could be helpful for the treatment of obesity, as proven previously by in vivo models. The aim of the present study is to determine if the supplementation with Lippia citriodora and Hibiscus sabdariffa is useful for the treatment of obesity and/or weight control in subjects without a controlled diet. The intake of the extract for 84 days reduced body weight, the body mass index, and the fat mass measured with both bioimpedance and densitometry. This decrease in fat mass was observed to a greater extent, being significant, in the fat mass of the trunk (chest and torso).

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OBJECTIVES: Regarding Lemon verbena gastroprotective effects, we investigated the protective effects of Lemon verbena extracts on reducing gastric ulcer induced by indomethacin. MATERIALS AND METHODS: Rats received aqueous and ethanolic extracts of Lemon verbena (50, 100, and 200 mg/kg), zileuton (100 mg/kg), montelukast (10 mg/kg), or 1% Tween 80 in presence or absence of indomethacin (100 mg/kg). RESULTS: Indomethacin produced stomach ulcer and increased neutrophils percentage and MDA level compared with the control group (P<0.001). Co-administration of indomethacin and zileuton, montelukast and ethanolic (200 mg/kg) (P<0.001), aqueous extract (200 mg/kg) (P<0.05) reduced ulcer compared with the indomethacin group (P<0.001). Ethanolic extracts (100 and 200 mg/kg) and aqueous extract (200 mg/kg) reduced the MDA level (P<0.001). Ethanolic (50, 100, and 200 mg/kg) and aqueous extracts (200 mg/kg) significantly decreased neutrophils percentage compared with the indomethacin group (P<0.001). CONCLUSION: Aqueous and particularly ethanolic extracts of Lemon verbena have protective effects on indomethacin-induced gastric ulcers.

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Lippia citriodora (LC) represents a complex plant-derived source of polyphenols and iridoids that has shown beneficial properties against obesity-related metabolic disorders. The complete extract and its major compound, verbascoside, have shown AMPK-activating capacity in cell and animal models. In this work, we aimed to elucidate the contribution of the different compounds present in the LC extract on the AMPK activation capacity of the whole extract. Semipreparative reversed-phase high-performance liquid chromatography coupled to electrospray ionization time-of-flight mass spectrometry (RP-HPLC-ESI-TOF-MS) was used to identify the major compounds with bioassay-guided fractionation in an adipocyte cell model for the measurement of AMPK activity. Twenty-two compounds were identified and purified almost to homogeneity in 16 fractions, and three compounds, namely verbascoside, luteolin-7-diglucuronide and loganic acid, showed the highest AMPK-activating capacity. The synergy study using the checkerboard and fractional inhibitory concentration index (FICI) methods exhibited synergistic behavior between loganic acid and luteolin-7-diglucuronide. Molecular docking experiments revealed that these three compounds might act as direct agonists of AMPK, binding to the AMP binding sites of the gamma subunit and/or the different sites of the interaction zones between the gamma and beta subunits. Although our findings conclude that the bioactivity of the extract is mainly due to verbascoside, the synergy found between loganic acid and luteolin-7-diglucuronide deserves further research aimed to develop optimized combinations of polyphenols as a new nutritional strategy against obesity-related metabolic disorders.

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Greener way of synthesizing nanoparticles has emerged as a substitute method, as it is ecological and cost effectual. Numerous efforts have been completed for green synthesis of silver oxide nanoparticles (Ag2O NPs) by various plant extracts. Current work disclosed the green combustion synthesis of Ag2O NPs by using Lippia citriodora plant powder. Furthermore, photocatalytic properties of Ag2O NPs were studied on acid orange 8(AO8) dye was assessed under UV light irradiation. The catalyst shows good photocatalytic activity (PCA) for the degradation of AO8 dye, NPs synthesized by Lippia citriodora powder shows high percentage of degradation. The Ag2O NPs act as excellent antibacterial against S. Aureus and antifungal activity against A. Aureus. Further wound healing studies in excision skin wound model in albino wistar rats showed the effective wound healing activity of Ag2O NPs incorporated hydrogels compared to untreated and plant extract treatments. The majority upshot of this research will be recommended that biologically synthesized Ag2O from Lippia citriodora plant powder has more valuable against various disease-causing pathogens and hence could be useful for developing wound dressing agents for nursing care.

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Lippia citriodora ethanolic extract (VEE) and verbascoside (Vs), a phenypropanoid glycoside, have been demonstrated to exert relaxant and anxiolytic properties. However, the molecular mechanisms behind their effects are still unclear. In this work, we studied the effects and action mechanisms of VEE and Vs in vivo and in vitro, on human neurotypic SH-SY5Y cells.TST was conducted on mice treated orally with VEE (25, 50 and 100 mg/Kg), Vs (2.5 and 5 mg/Kg), Bupropion (20 mg/Kg) and Milli-Q water. Higher dose of VEE-treated mice showed an increase of immobility time compared to control groups, indicating an induction of relaxation. This effect was found to be induced by regulation of genes playing key roles in calcium homeostasis (calcium channels), cyclic AMP (cAMP) production and energy metabolism. On the other hand, low doses of VEE and Vs showed an antidepressant-like effect and was confirmed by serotonin, noradrenalin, dopamine and BDNF expressions. Finally, VEE and Vsenhancedcell viability, mitochondrial activity and calcium uptake in vitro confirming in vivo findings. Our results showed induction of relaxation and antidepressant-like effects depending on the administered dose of VEE and Vs, through modulation of cAMP and calcium.

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