Attenuation of cannabis withdrawal symptoms by Prosopis farcta extract, its luteolin and melatonin in mice: Involvement of brain-derived neurotrophic factor and dopamine.

The aim of this study was the identification of luteolin in Prosopis farcta extract (PFE) and melatonin to evaluate its effect on THC withdrawal syndrome in mice. Luteolin was identified by high-performance liquid chromatography (HPCL). Signs of toxicity of mice in PFE and luteolin were monitored for LD50 calculation. The behavioral symptoms of THC withdrawal (stereotypies, ambulation, and inactivity time) induced by the rimonabant challenge were illustrated in THC-dependent mice receiving PFE, luteolin, and melatonin. The expression of mature BDNF (mBDNF) was evaluated by Western blot analysis. The dopamine concentrations were measured using HPLC. PFE and luteolin LD50 were 650 and 220 mg/kg, respectively. PFE (300 mg/kg), all doses of luteolin, and melatonin increased significantly the mBDNF expression and decreased the dopamine concentration. The findings suggest that PFE, luteolin, and melatonin are mighty in reducing the signs of THC withdrawal. It seems these effects were due to a decrease in dopamine concentration level and an increase in mBDNF protein expression in mice brains.

Biological Activities of Zinc Oxide Nanoparticles Green Synthesized Using the Aqueous Extract of Dracocephalum kotschyi Boiss.

BACKGROUND: Dracocephalum kotschyi Boiss. is known as a native medicinal plant of Iran. OBJECTIVE: In this study, aqueous extract of D. kotschyi was used to synthesize ZnO-NPs. To produce ZnO-NPs, aerial parts of D. kotschyi were powdered and then macerated for obtaining aqueous extract, after that, aqueous extract was used to reduse zinc nitrate to ZnO-NPs. METHODS: To confirm nanoparticles synthesis, SEM, TEM, UV-Vis, FTIR, and XRD were used. The synthesized ZnO-NPs were studied for antimicrobial activities by microdilution method for calculating MIC and MBC. Analysis of ZnO-NPs confirmed successful synthesis by extract of D. kotschyi. RESULTS: The sizes of ZnO-NPs were estimated 50-200 nm in diameter. Antibacterial and antifungal experiments showed potent activities against Staphylococos aureus, Pseudomonas aeruginosa and Candida albicans. The results of the studies showed that the nanoparticles synthesized with the aqueous extract of D. kotschyi have a much greater antimicrobial effect than the aqueous extract of D. kotschyi and zinc nanoparticles, each alone (MIC values 3.7 to 7.5 mg/ml). CONCLUSION: The noteworthy point is that the inhibitory rate of synthesized zinc oxide nanoparticles is higher compared to broad-spectrum antibiotics, such as chloramphenicol (MIC values 15 mg/ml). Determining the therapeutic and toxic dose of this product for humans requires further investigation and clinical trials.

Immune-enhancing activity of Astragalus maximus extract for inhibiting the Toxoplasma gondii infection: experimental research.

Background: This study aimed to evaluate the in-vitro anti-Toxoplasma effects and cytotoxicity effects of Astragalus maximus chloroformic extract (AMCE) on the T. gondii Rh strain. Methods: In-vitro effects of AMCE (2-64 µg/ml) on tachyzoites were measured by MTT assay for 48 h. The effects of AMCE on infectivity rate and intracellular parasites into macrophage cells (J774-A1) were evaluated. The Griess reaction assay and quantitative real-time PCR were used to determine the nitric oxide (NO) and the mRNA expression levels of IFN-γ and iNOS in infected J774-A1 macrophage cells. Results: The mortality rate of the parasites significantly (P<0.001) increased in a dose-dependent manner with an IC50 value of 9.85 µg/ml. The rate of infection and the mean number of intracellular tachyzoites in macrophage cells were significantly reduced (P<0.001) after exposure of the macrophage cells to AMCE. The mRNA expression levels of IFN-γ, iNOS, and NO production in macrophage cells after treatment with the AMCE were increased, especially at the concentration of ½ IC50 and IC50 (P<0.001). Conclusion: The findings of the current in vitro investigation revealed favorable anti-Toxoplasma effects of AMCE against tachyzoites and intracellular forms of T. gondii. Despite the fact that the accurate anti-Toxoplasma mechanisms of AMCE are not clear, our results showed that triggering NO production and cellular immunity can be considered as the main mechanisms of action of AMCE for controlling and eliminating T. gondii. However, further surveys are mandatory to assess the efficacy and safety of AMCE in an animal model and its accurate mechanisms of action.

Antileishmanial, cellular mechanisms, and cytotoxic effects of green synthesized zinc nanoparticles alone and in combined with glucantime against Leishmania major infection.

BACKGROUND: We decided to investigate the antileishmanial, cellular mechanisms, and cytotoxic effects of green synthesized Zinc nanoparticles (ZnNPs) alone and combined with glucantime against Leishmania major infection. METHODS: The effect of green synthesized ZnNP on L. major amastigote was studied through macrophage cells. The mRNA expression level of iNOS and IFN-γ followed by the exposure of J774-A1 macrophage cells to ZnNPs was assessed by Real-time PCR. The Caspase-3-like activity of promastigotes exposed to ZnNPs was studied. Effects of ZnNPs alone and combined with glucantime (MA) were studied on cutaneous leishmaniasis in BALB/c mice. RESULTS: ZnNPs displayed the spherical shape with sizes ranging from 30 to 80 nm. The obtained IC50 values for ZnNPs, MA, and ZnNPs + MA were 43.2, 26.3, and 12.6 µg/mL, respectively; indicating the synergistic effects of ZnNPs in combination with MA. CL lesions had completely improved in the mice received with ZnNPs in combination with MA. The mRNA expression level of iNOS, TNF-α, and IFN-γ was dose-dependently (p < 0.01) upregulated; whereas it was downregulated in IL-10. ZnNPs markedly stimulated the caspase-3 activation with no significant toxicity on normal cells. CONCLUSION: Based on these in vitro and in vivo results, green synthesized ZnNPs, mainly along with MA, showed that has the potential to be introduced as a new drug for CL therapy. Triggering of NO production, and inhibition of infectivity rate are revealed as mechanisms of action ZnNPs on L. major. But, supplementary investigations are necessary to clear the efficacy and safety of these agents.

Green Synthesis, Characterization, and Antiparasitic Effects of Gold Nanoparticles against Echinococcus granulosus Protoscoleces.

Echinococcosis, or hydatidosis, is one of the most important zoonotic diseases, which is initiated by the larval stage in the clasts of Echinococcus granulosus. For the treatment of hydatidosis, surgery is still the preferred method and the first line of treatment for symptomatic patients. Unfortunately, most of the scolicidal agents that are injected inside cysts during hydatid cyst surgery have side effects, including leaking out of the cyst and adverse effects on the living tissue of the host, such as necrosis of liver cells, which limits their use. This work was carried out to study the lethal effect of green synthesized gold nanoparticles (Au-NCs) against hydatid cyst protoscoleces. Au-NCs were green synthesized using the Saturja khuzestanica extract. Au-NCs were characterized by UV-visible absorbance assay, electron microscopy analysis, X-ray diffraction (XRD), and Fourier transform infrared (FTIR) spectroscopy. Scolicidal properties of Au-NCs (1-5 mg/mL) were studied against protoscoleces for 10-60 min. The effect of Au-NCs on the expression level of the caspase-3 gene as well as the ultrastructural examination was studied by real-time PCR and scanning electron microscopy (SEM). The cytotoxicity of Au-NCs on hepatocellular carcinoma (HepG2) and normal embryonic kidney (HEK293) cell lines was also studied by the cell viability assay. The obtained Au-NCs are cubes and have an average size of 20-30 nm. The highest scolicidal efficacy was observed at 5 mg/mL with 100% mortality after 20 min of treatment for hydatid cyst protoscoleces. In ex vivo, Au-NCs required more incubation time, indicating more protoscolicidal effects. Au-NCs markedly upregulated the gene level of caspase-3 in protoscoleces; whereas they changed the ultra-structure of protoscoleces by weakening and disintegrating the cell wall, wrinkles, and protrusions due to the formation of blebs. We showed the effective in vitro and ex vivo scolicidal effects of Au-NCs against hydatid cyst protoscoleces by provoking the apoptosis process of caspase-3 activation and changing the ultrastructure of protoscoleces with no significant cytotoxicity against human normal cells. However, additional studies should be conducted to determine the possible harmful side effects and accurate efficacy.

Leishmanicidal and immunomodulatory activities of the formononetin (a natural isoflavone) against Leishmania tropica.

OBJECTIVE: This work aimed to examine the leishmanicidal, cellular mechanisms and cytotoxicity effects of formononetin (FMN), a natural isoflavone, against Leishmania tropica. We used the MTT assay to determine the leishmanicidal effects of FMN against promastigotes and its cytotoxicity effects on J774-A1 macrophage cells. The Griess reaction assay and quantitative real-time PCR were used to determine the nitric oxide (NO) and the mRNA expression levels of IFN-γ and iNOS in infected J774-A1 macrophage cells. RESULTS: FMN significantly (P < 0.001) decreased the viability and number of promastigotes and amastigotes forms. The 50% inhibitory concentrations value for FMN and glucantime was 9.3 and 14.3 µM for promastigote and amastigote, respectively. We found that the macrophages exposed with FMN especially at concentrations of 1/2 IC50 and IC50 significantly activated the NO release and the mRNA expression levels of IFN-γ, iNOS. The findings of the current research showed the favorable antileishmanial effects formononetin, a natural isoflavone, against various stages of L. tropica through inhibition of infectivity rate of macrophage cells and triggering the NO production and cellular immunity. However, supplementary works are essential to evaluate the ability and safety of FMN in animal model before use in the clinical phase.

High potency of magnetic iron oxide nanoparticles covered by piroctone olamine against cystic echinococcosis.

This study examined the synthesis of magnetic iron oxide nanoparticles coated with PO (FOMNPsP) and assessed their in vitro, ex vivo, and in vivo effects against cystic echinococcosis. The FOMNPsP was synthesized through the alkalization of iron ions in a deoxygenated form. In vitro and ex vivo protoscolicidal effects of FOMNPsP (100-400 µg/mL) were evaluated on hydatid cyst protoscoleces by the eosin exclusion test for 10-60 min. The effect of FOMNPsP on caspase-3 gene expression and exterior ultra-structural of protoscoleces was assessed by real-time PCR and scanning electron microscopy (SEM), respectively. In vivo effects were assessed by evaluating the number, size, and weight of hydatid cysts among infected mice. The FOMNPsP size was < 55 nm, and the most frequent particles were in the 15-20 nm range. In vitro and ex vivo assays revealed that the highest protoscolicidal effect was observed at 400 µg/mL with 100% lethality. After exposure of protoscoleces with FOMNPsP, the level of gene expression of caspase-3 was dose-dependently increased (p < 0.05). By SEM, the FOMNPsP-treated protoscoleces showed wrinkles and bulges resulting from the formation of blebs. FOMNPsP significantly decreased (p < 0.01) the mean number, size, and weight of the hydatid cyst. FOMNPsP revealed the potent protoscolicidal traits through disrupting the cell wall and apoptosis induction. The results also indicated the promising effect of FOMNPsP in controlling hydatid cysts in the animal model. Although FOMNPsP is safe for human normal cells, more investigations are required to clarify its toxicity and precise mechanisms of action.

In-vivo and in vitro assessments of the radioprotective potential natural and chemical compounds: a review.

PURPOSE: The study of the radioactive role of natural and chemical substances on human and animal studies has been the subject of research by some researchers. Therefore, the review of some of the past and current studies conducted in this field, can provide helpful information to elucidate of the importance of radioprotective components in reducing radiation exposure side effects. METHODS: The authors search for keywords including In vitro, In vivo, Radioprotective, Ionizing radiation, and Vitamin in ScienceDirect, Scopus, Pubmed, and Google Scholar databases to access previously published articles and search for more reference articles on the role of radioprotective materials from natural and chemical compounds. RESULTS: Radiation exposure can produce reactive oxygen species (ROS) in the body, however most of which are eliminated by the body's natural mechanisms, but when the body's antioxidant systems do not have enough ability to neutralize free radicals, oxidative stress occurs, which causes damage to DNA and body tissues. Therefore, it is necessary use of alternative substances that reduce and inhibit free radicals. CONCLUSION: In general, recommended that antioxidant component(s) can be protect tissue damages in humans or animals, due to the their ability to scavenge free radicals generated by ionizing radiation.

Anti-helminthic effects and cellular mechanisms of Astragalus ecbatanus extract against Echinococcus granulosus protoscoleces.

We aimed to investigate the in vitro and ex vivo anti-helminthic effects of Astragalus ecbatanus chloroform extract (AECE) as well as its cellular mechanisms against Echinococcus granulosus protoscoleces. In vitro and ex vivo anti-helminthic effects of AECE on E. granulosus protoscoleces were evaluated through eosin exclusion test. Effects of AECE on induction of the caspase-3 like activity, the permeability of plasma membrane were also determined. The effects of AECE against the human hepatocyte carcinoma (HepG2) and human embryonic kidney (HEK293T) cells were determined by MTT ([3-(4.5-dimethylthiazol-2-yl)-2.5-diphenyl tetrazolium bromide)]) test. The findings revealed that AECE especially at 45 mg/mL displayed potent in vitro and ex vivo anti-helminthic effects against E. granulosus protoscoleces (p < 0.001). After treatment of protoscoleces with AECE, the caspase-3 enzyme activity and the rate of permeability of plasma membrane was dose dependently (P < 0.001) increased. The 50% cytotoxic concentration (CC50) value of AECE for normal and cancer cells lines 4.62 mg/mL and 1.89 mg/mL, respectively. The findings of the current study revealed the favorable in vitro and ex vivo anti-helminthic effects of AECE against protoscoleces of E. granulosus. However, further surveys in animal model are required to elucidate different aspects of this extract before use in clinical phases.

Chemical composition, antileishmanial, and cytotoxic effects Ferula macrecolea essential oil against Leishmania tropica.

Background: The current study was aimed to evaluate the chemical composition, as well as antileishmanial and cytotoxic effects of the essential oil of Ferula macrecolea and its main compound, terpinolene, against promastigotes and amastigotes of Leishmania tropica. Methods: The chemical composition of the essential oil was analyzed by a gas chromatograph connected to a mass spectrometer (GC/MS). The MTT (3-(4.5-dimethylthiazol-2-yl) - 2.5-diphenyl tetrazolium bromide) assay was used to study the effects of the essential oil and terpinolene against promastigotes while the macrophage model was used for evaluating the effect of F. macrecolea essential oil against amastigotes of L. tropica as well as assessing cytotoxicity. The Griess reaction assay was employed to study the nitric oxide (NO) produced by treating macrophage cells with the essential oil and terpinolene. Furthermore, the effect of the essential oil and terpinolene on plasma membrane permeability and inhibition of infection in macrophages was evaluated. Results: The main compounds were terpinolene (77.72%), n-nonanal (4.47%), and linalool (4.35%), respectively. The 50% inhibitory concentrations (IC50) of the essential oil, terpinolene, and glucantime against promastigotes were 27.6, 11.6, and 32.8 µg/mL, respectively; however, their IC50 values against amastigotes were 42.3, 19.6, and 56.9 µg/mL, respectively. The 50% cytotoxic concentrations of the essential oil, terpinolene, and glucantime were 471.3, 207.3, and 1165.3 µg/mL, respectively. The production of NO in macrophage cells after treatment with the essential oil and terpinolene was increased in a dose-dependent manner (p < 0.001). The results revealed that by increasing the concentration of the essential oil and terpinolene, the permeability of the parasites' plasma membrane was significantly changed (p < 0.001). The pre-incubation of Leishmania parasites with F. macrecolea essential oil and terpinolene significantly declined the rate of cell infection by 74.8% and 79.4%, respectively (p < 0.001). Conclusion: The results of the present study indicated that F. macrecolea essential oil, especially its main compound, i.e., terpinolene, has a potent antiparasitic effect on the promastigote and amastigote stages of L. tropica. Considering the advantages of medicinal plant products over their chemical counterparts, it is suggested that in the continuation of this study, the effect of F. macrecolea essential oil, especially terpinolene, on laboratory animals, and in case of high efficiency, in humans be evaluated.

Ceratonia siliqua L improved cryodamage of asthenozoospermic specimens: An experimental study.

Background: Sperm freezing is an important procedure in assisted reproductive technology. Freezing results in physical and chemical changes in the sperm. Ceratonia siliqua L (C.siliqua) is a tree that has antioxidant properties. Objective: This study aimed to investigate the effect of different concentrations of C.siliqua in a freezing medium on semen parameters, and some biochemical parameters in asthenozoospermic specimens. Materials and Methods: Forty asthenozoospermic specimens (semen specimens with motility < 32%) were obtained from men aged between 20-40 yr according to the World Health Organization criteria. Each sample was divided into 6 groups: I) fresh, II) control, III) 5, IV) 10, V) 20, and VI) 30 µg/ml C.siliqua extract were added to a freezing medium respectively. Then sperm parameters, malondialdehyde, total antioxidant capacity, reactive oxygen species, and sperm DNA assay were evaluated using related protocols after thawing. Results: Data analysis shows that sperm parameter, and total antioxidant capacity level increased at a concentration of 20 µg/ml of C.siliqua extract compared to the other concentrations of C.siliqua extract after cryopreservation and thawing (p < 0.001). Also, the sperm DNA fragmentation assay, reactive oxygen species, and malondialdehyde levels were significantly reduced by adding 20 µg/ml of C.siliqua extract to the sperm freezing medium compared to the other treated groups after cryopreservation (p < 0.001). Conclusion: C.siliqua extract significantly improved sperm parameters after cryopreservation and thawing in asthenozoospermic specimens, and the greatest impact was observed at the 20 µg/ml C.siliqua L extract concentration (p < 0.001).

Evaluation of Phytochemical, Antioxidant and Antibacterial Activity on Astragalus Chrysostachys Boiss. Roots.

Astragalus is a well-known genus in Leguminosae family that represented more than 800 species growing in Iran. Nevertheless, there are a few reports on Astragalus plants endemic to Iran. The roots of Astragalus plants are rich in saponins, flavonoids and polysaccharides that possess various pharmacological activities. In the present study, chemical components, antioxidant and antibacterial activity of Astragalus chrysostachys Boiss. roots were evaluated. For determination of phytochemicals in Astragalus chrysostachys Boiss. roots, total hydroalcoholic extract was fractionated with ethyl acetate and n-butanol. Ethyl acetate extract as a flavonoid rich extract was analyzed using vacuum liquid chromatography and preparative TLC and consequently a major flavonoid was isolated. The structure of the obtained compound was elucidated with 1D and 2D NMR experiments. Additionally, the essential oil of the roots was analyzed by GC-MS. Antioxidant activity of all extracts was evaluated by different assays. Moreover, antibacterial activities of the extracts were also investigated against 2 Gram-positive and 2 Gram-negative bacteria using Micro-dilution Broth method. Apigenin-6, 8-di-C-glucoside was detected in ethyl acetate extract for the first time in genus Astragalus. In addition, m-tolualdehyde, acetophenone, croweacin were found to be characteristics of the volatile oil of roots. Ethyl acetate extracts revealed notable antioxidant activity in DPPH scavenging assay with IC50 value of 14.6 µg/mL. Evaluation of antibacterial activity on the tested extracts showed mild activity against Gram-positive bacteria. Since there have been no reports on Astragalus chrysostachys Boiss. to date, the present data might be promising for application of this plant derivatives in phytotherapeutic practice.

Chemical composition, antioxidant activity and antibacterial effect of essential oil of the aerial parts of Salvia sclareoides.

OBJECTIVE: To determine the chemical composition, antioxidant activity and antibacterial properties of essential oils of the aerial parts of Salvia sclareoides (S. sclareoides). METHODS: The essential oil of the areal parts of S. sclareoides was obtained by using hydrodistillation method and the composition of the volatile components analyzed by gas chromatography method coupled with mass spectrometry detector. The antimicrobial capacity of the essential oil of S. sclareoides was investigated by microdilution technique. The antioxidant activities were determined employing inhibition of 2,2-diphenyl-1-picrylhydrazyl hydrate radical method. RESULTS: Mass spectra were searched against mass spectrometry databases and sixty components were recognized. Non-terpenoids (41.6%) and sesquiterpenes (39.7%) were determined as the main components of the essential oil. The main identified components were, linalool (27.6%), trans-caryophyllene (16.6%), beta.-trans-ocimene (11.831%), germacrene-D (10%), bicyclogermacrene (3.3%) and caryophyllene oxide (2.8%). Two monoterpens (13.2%) and three oxygenated sesquiterpenes (5.5%) were also obtained from the essential oil of the S. sclareoides CONCLUSIONS: Results indicated that essential oil of S. sclareoides includes rather higher proportions of non-terpenoid and sesquiterpenes compounds with good antioxidant and antibacterial properties.

Plasma stable, pH-sensitive fusogenic polymer-modified liposomes: A promising carrier for mitoxantrone.

pH-sensitive liposomes are designed to undergo acid-triggered destabilization. In the present study, we prepared polymer-modified, plasma stable, pH-sensitive fusogenic mitoxantrone liposomes to increase efficacy and selectivity on cancer cell lines. Conventional liposomes were prepared using cholesterol and dipalmitoyl-sn-glycero-3-phosphatidylethanolamine. Dioleoylphosphatidylethanolamine and a cholesteryl derivative, poly(monomethylitaconate)-co-poly(N,N-dimethylaminoethyl methacrylate) (PMMI-co-PDMAEMA), were used for the preparation of pH-sensitive fusogenic liposomes. Using polyethylene glycol (PEG)-poly(monomethylitaconate)-CholC6 (PEG-PMMI-CholC6) copolymers instead of cholesterol introduced pH-sensitive and plasma stability properties simultaneously in prepared liposomes. All formulations were prepared by thin film hydration method and subsequently, pH-sensitivity and stability in human serum were evaluated. The ability of pH-sensitive fusogenic liposomes to enhance the mitoxantrone cytotoxicity and selectivity in cancerous cell lines was assessed in vitro compared to normal cell line using human breast cancer cell line (MCF-7), human prostate cancer cell line (PC-3), and human umbilical vein endothelial cells line. Results revealed that both PMMI-co-PDMAEMA and PEG-PMMI-CholC6-based formulations showed pH-sensitive property and were found to rapidly release mitoxantrone under mildly acidic conditions. Nevertheless, only the PEG-PMMI-CholC6-based liposomes preserved pH-sensitivity after incubation in plasma. Mitoxantrone loaded-pH-sensitive fusogenic liposomes exhibited a higher cytotoxicity than the control conventional liposomes on MCF-7 and PC-3 cell lines. On the contrary, both pH-sensitive fusogenic liposomes showed lower cytotoxic effect on human umbilical vein endothelial cell line. Plasma stable, pH-sensitive fusogenic liposomes are promising carriers for enhancing the efficiency and selectivity, besides reduction of the side effects of anticancer agents.