Comparison effects of Ferula gummosa essential oil and Beta-pinene Alginate nanoparticles on human melanoma and breast cancer cells proliferation and apoptotic index in short term normobaric hyperoxic model.

BACKGROUND: Breast cancer is the most common cancer among women, and melanoma is the most dreadful type of skin cancer. Due to the side effects of chemotherapy drugs, the development of new herbal nano-medicines has been considered. METHODS: This study first investigated the chemical composition of Ferula gummosa essential oil using GC-MS analysis; ß-pinene, with 61.57%, was the major compound. Next, alginate nanoparticles containing ß-pinene and the essential oil with particle sizes of 174 ± 7 and 137 ± 6 nm were prepared. Meanwhile, their zeta potentials were 12.4 ± 0.7 and 28.1 ± 1 mV. Besides, the successful loading of ß-pinene and the essential oil in nanoparticles was confirmed using ATR-FTIR analysis. After that, their effects on viability and apoptotic index of human melanoma and breast cancer cells were investigated in normoxia and normobaric hyperoxia (NBO) conditions. RESULTS: The best efficacy on A-375 and MDA-MB-231 cells was achieved by alginate nanoparticles containing the EO at hyperoxic and normoxia conditions; IC50 76 and 104 µg/mL. Besides, it affected apoptosis-involved genes; as Bax/Bcl-2 ratio was higher than 1, conditions for induction of apoptosis were obtained. Higher sensitivity was observed in the A-375 cell line treated with Alg-EO in the NBO model. CONCLUSIONS: Alginate nanoparticles containing F. gummosa EO could be considered for further investigation in anticancer studies. Also, it may be expected that NBO can be a new strategy for delaying cancer progression and improving nanotherapy efficacy.

Tissue engineering studies in male infertility disorder.

Infertility is an important issue among couples worldwide which is caused by a variety of complex diseases. Male infertility is a problem in 7% of all men. In vitro spermatogenesis (IVS) is the experimental approach that has been developed for mimicking seminiferous tubules-like functional structures in vitro. Currently, various researchers are interested in finding and developing a microenvironmental condition or a bioartificial testis applied for fertility restoration via gamete production in vitro. The tissue engineering (TE) has developed new approaches to treat male fertility preservation through development of functional male germ cells. This makes TE a possible future strategy for restoration of male fertility. Although 3D culture systems supply the perception of the effect of cellular interactions in the process of spermatogenesis, formation of a native gradient of autocrine/paracrine factors in 3D culture systems have not been considered. These results collectively suggest that maintaining the microenvironment of testicular cells even in the form of a 3D-culture system is crucial in achieving spermatogenesis ex vivo. It is also possible to engineer the testicular structures using biomaterials to provide a supporting scaffold for somatic and stem cells. The insemination of these cells with GFs is possible for temporally and spatially adjusted release to mimic the microenvironment of the in situ seminiferous epithelium. This review focuses on recent studies and advances in the application of TE strategies to cell-tissue culture on synthetic or natural scaffolds supplemented with growth factors.

Probiotic bacteria alleviate chlorpyrifos-induced rat testicular and renal toxicity: A possible mechanism based on antioxidant and anti-inflammatory activity.

Chlorpyrifos (CPF) has caused many potential toxicities in nontarget organisms. Fewer studies have been conducted on the effects of lactic acid bacteria (LAB) in mitigating tissue damage induced by CPF in vivo. Therefore, we investigated CPF renal and testicular toxicity and the alleviating effect of probiotic lactobacilli, based on antioxidant and anti-inflammatory activity, on induced toxicity in an animal model. Biochemical assays showed that CPF induced oxidative stress along with a change in superoxide dismutase (SOD) and catalase (CAT) activity in a tissue-dependent manner. After treatment with CPF, testicular and renal levels of TNF-α were significantly reduced and enhanced, respectively, compared to the control group. The probiotic treatment restored renal and testicular TNF-α levels and modulated and blocked the increasing effect of CPF on renal IL-1ß levels. Testicular IL-1ß levels in the probiotic-treated and CPF groups demonstrated similar values. Exposure to CPF significantly induced renal histopathological damage that, of course, was completely inhibited by treatment with Lactobacillus casei and the LAB mixture. In summary, CPF showed significant toxicological effects on oxidative stress and the inflammation rate in CPF-exposed rats. Therefore, supplementation with probiotic bacteria may alleviate CPF renal toxicity and mitigate its oxidative stress and inflammation effects.

Evaluation of the topical gel and oral administration of Punica Granatum Var Pleniflora on oral mucositis induced by 5-Fluorouracil in golden hamsters.

BACKGROUND: Oral mucositis (OM), an acute inflammation of the oral cavity, is a common complication in patients undergoing invasive myeloblastic chemotherapy or radiation therapy. 5-fluorouracil (5-FU) is one of the most effective therapeutic drugs, but one of the common side effects of 5-FU administration is OM. Unfortunately, no suitable treatment has been found, so far to control its side effects. Studies showed that herbal medicine like Punica granatum var pleniflora (PGP) has medicinal properties such as anti-inflammatory and antibacterial and can be an alternative for the treatment of fungal infection. Accordingly, we decided to investigate the therapeutic effect of PGP in the treatment of OM caused by 5-FU in golden hamsters. METHODS: Sixty male golden hamsters were divided into six main group. Chemotherapy with 5-FU at dose of 60 mg/kg was performed at a ten-day duration. Then, cheek pouches of the hamsters were scratched with an 18-gauge sterile needle to induce oral mucositis in animals. On the twelfth day, as a day of intensification of OM, treatment with PGP including topical gel with concentrations of 5% and 10% and oral administration of hydro-alcoholic extract with doses of 125 mg/kg and 250 mg/kg for three- and five-day therapeutic duration were separately started. Finally, samples of cheek pouches in hamsters were collected on 14th and 17th days and histopathologic score (HPS), malondialdehyde (MDA), and myeloperoxidase (MPO) levels were assayed. RESULTS: A significant (p < 0.05) decrease in histopathologic score was observed in G10%-, P125-treated groups in comparison to the Ctrl group. Our data showed that treatment with G10% is more potent than P125-treated group. In contrast, histopathologic score in G10%, P125, and P250 treated groups demonstrated almost similar values On the 17th day. However, the levels of MDA and MPO in the treatment groups were enhanced compared with control group (p < 0.05). CONCLUSIONS: It is possible that PGP can play protective role in the healing of tissue damage caused by chemotherapy with 5-FU due to the presence of its natural compounds and antioxidant properties.

Attenuating NF-κB/VCAM-1 Expression in the Middle Cerebral Artery Occlusion Model by Viola Odorata: Protection Against Ischemia-Reperfusion Injury in Rats.

Introduction: The death of neurons and cerebral edema are the main consequences of stroke. However, inflammatory processes play key roles in aggravating cerebral damage following a stroke. This study aimed to investigate the effects of Viola odorata extract (VOE) on the infarct volume (IV), neurologic deficits (ND), and the expression of NF-κB and VCAM-1 in the Middle Cerebral Artery Occlusion (MCAO) model. Methods: The animals were randomly separated into 6 groups: (1) control group, (2) vehicle-treated group, (3) MCAO group, (4) VOE25 group, (5) VOE50 group, and (6) VOE75 group (n= 12). VOE (25, 50, and 75 mg/kg) and distilled water were administered daily for 30 days. Two hours after the last gavage, the rats were exposed to MCAO for 60 minutes. Twenty-four hours later, IV, ND, and NF-κB/VCAM-1 expressions were evaluated. Results: Viola odorata extract exhibited excellent neuroprotective effects by reducing IV (mainly in the core and subcortex areas), and induced downregulation of NF-κB and VCAM-1 expression. Conclusion: Viola odorata could also activate intracellular pathways, reducing the expression of NF-κB and VCAM-1. It is useful for developing a novel medical herb for treating cerebral ischemia. Highlights: A stroke occurs when the blood supply to a part of the brain is interrupted or reduced.Viola odorata extract (VOE) reduced the infarct volumes (IV) in rats' brains 24 h after middle cerebral artery occlusion (MCAO).VOE may decrease IV in the MCAO model by downregulating the NF-κB/VCAM-1 expression. Plain Language Summary: A stroke is a medical condition in which poor blood flow to the brain results in cell death. After an interruption of blood flow, energy stores are rapidly depleted, and complex cellular cascades induce excitotoxic cell death. So far, no effective and specific treatment has been suggested for the stroke. In this study, the neuroprotective effects of the viola odorata extract (VOE) were screened. The V. odorata extract exhibited excellent neuroprotective effects by reducing infarct volume and inducing downregulation of NF-κB/VCAM-1 expression. This finding suggests that V. odorata may be useful for developing a novel medical herb for treating cerebral ischemia.

Nanoemulsion and Nanogel Containing Eucalyptus globulus Essential Oil; Larvicidal Activity and Antibacterial Properties.

Eucalyptus globulus essential oil (EGEO) possesses many biological effects such as antibacterial, antifungal, and insecticide properties. In the current study, the chemical composition of EGEO was first investigated using GC-MS analysis. Then, a nanoemulsion and nanogel containing EGEO (EGEO-nanoemulsion and EGEO-nanogel) were prepared. After that, the successful loading of EGEO was confirmed using ATR-FTIR analysis. EGEO-nanoemulsion and EGEO-nanogel with LC50 values of 27 and 32 µg/mL showed promising efficacies against Anopheles stephensi larvae. Besides, the efficacy of EGEO-nanogel (IC50 187 µg/mL) was significantly more potent than EGEO-nanoemulsion (IC50 3732 µg/mL) against Staphylococcus aureus. However, no significant difference was observed in the efficacy of EGEO-nanoemulsion and EGEO-nanogel against Pseudomonas aeruginosa. Natural components, straightforward preparation, and proper efficacy are some of the advantages of EGEO-nanogel; it could be considered for further consideration against other pathogens and mosquito larvae.

Anti-inflammatory and anti-nociceptive effects of Cinnamon and Clove essential oils nanogels: an in vivo study.

BACKGROUND: Cinnamon (Cinnamomum zeylanicum) and Clove (Syzygium aromaticum) essential oils are two medicinally important plant-derived substances with a wide range of biological properties. Besides, nanoemulsion-based gels have been widely used to increase topical drug delivery and effectiveness. METHODS: This study aimed to explore the anti-inflammatory effect (paw edema test) and the anti-nociceptive effect (hot plate and formalin test) of nanoemulsion-based gels containing the essential oils in the animal model. Cinnamon and Clove essential oils nanoemulsions with droplet sizes of 28 ± 6 nm and 12 ± 3 nm were first prepared. By adding carboxymethylcellulose (3.5% w/v), the nanoemulsions were then gelified. Finally, the nanogels were characterized by ATR-FTIR analysis and were used as topical pre-treatment before induction of inflammation or pain in acute and chronic analgesic experimental studies. RESULTS: The paw edema and formalin findings showed that the nanogels formulations possess significant anti-nociceptive and anti-inflammatory effects. CONCLUSION: The prepared nanogels could be considered as analgesic drugs for inhibiting the inflammation and pain of diseases.

Subacute toxicity of chlorpyrifos on histopathological damages, antioxidant activity, and pro-inflammatory cytokines in the rat model.

Chlorpyrifos (CPF) is an extensively used organophosphorus pesticide for agricultural, industrial, and domestic purposes. Previous studies have reported the adverse effects of CPF, such as intoxication incidents, endocrine disruption, cardiovascular diseases, as well as histopathological and oxidative damage. The aims of the present study were to elucidate short time subacute toxicity of CPF in male rats. Sprague-Dawley male rats (n = 32) were divided into four groups (n = 8) and received CPF as 3.25 mg/kg body weight (b.w) (Group A), 6.75 mg/kg b.w (Group B), 13.5 mg/kg b.w (Group C), and corn oil (control or Group D) daily via gavage for 15 days. The rats were sacrificed and oxidative damages, pro-inflammatory cytokines (TNF-α, IL-1ß), and histopathological changes were determined in the lung, liver, kidney, heart, and testis tissues as well as plasma. According to our result, administration of CPF caused a significant increase in malondialdehid level and catalase activity while a significant decrease in superoxide dismutase activity in all tissues. In addition, a significant decrease in TNF-α observed in all tissues and plasma duo to the CPF. Histopathological evaluation of CPF-treated samples revealed a dose-dependent tissue toxicity in the liver, heart, lung, and kidney with less sensitivity of testicular and kidney tissues. These results suggest the potential of CPF in inducing oxidative stress at low doses and short duration time with similar trends in different tissues. As well as, due to the effects of CPF on some pro-inflammatory mediators, more comprehensive studies are recommended.

Psoralidin exerts anti-tumor, anti-angiogenic, and immunostimulatory activities in 4T1 tumor-bearing balb/c mice.

BACKGROUND: Psoralidin as a compound of the Psoralea corylifolia seeds exhibited several anti-cancer potentials in various cancers. MATERIALS AND METHODS: In this study, 4T1 tumor-bearing Balb/c mice were treated by intraperitoneal administration of Psoralidin, and Paraffin, as a control group to investigate anti-tumor, anti-angiogenic, and immunostimulatory activities in breast cancer. Body weight and tumor volume measurement were performed. Hematoxylin and Eosin (H&E) staining as well as immunohistochemistry for Ki-67, CD31 and VEGF markers were conducted. In addition, ELISA assay was performed for evaluating the serum level of IFN-γ and IL-4. Moreover, real time assay was performed to evaluate the expression of angiogenesis and immunostimulatory related genes. RESULTS: There were no significant changes in the body weight of all animal groups. The anti-cancer effects of Psoralidin were significantly observed after 24 days of the last treatment, confirmed by smaller tumor volume and also H&E staining. The expression level of Ki-67, CD31 and VEGF were significantly decreased in tumor tissues of the Psoralidin-treated group in comparison with Paraffin-treated group. Moreover, there was a significant reduction in the serum level of IL-4 in tumor-bearing mice after Psoralidin treatment while the serum level of IFN-γ was significantly augmented in all groups. Moreover, the reduction in expression of VEGF-a and IL-1ß was observed. Interestingly Psoralidin treatment led to expression increase of FOXp3. CONCLUSIONS: Psoralidin shows the anti-cancer potential in an animal model of breast cancer; however, further studies are recommended to elucidate its mechanisms of action.

Chitosan nanoparticles containing limonene and limonene-rich essential oils: potential phytotherapy agents for the treatment of melanoma and breast cancers.

BACKGROUND: Melanoma and breast cancers are two common cancers worldwide. Due to the side effects of chemotherapy drugs and the occurring resistance against them, the development of green drugs has been received more attention. METHODS: The anticancer effects of three essential oils from the Citrus family and their identified major constituents (limonene) were first investigated against melanoma and breast cancer cell lines (A-375 and MDA-MB-468). By preparing chitosan nanoparticles containing them, an attempt was then made to improve their effectiveness. RESULTS: Chitosan nanoparticles containing Citrus sinensis and Citrus limon essential oils with IC50s of 0.03 and 0.124 µg/mL on A-375 cells, and 23.65 and 40.32 µg/mL on MDA-MB-468 showed distinct anticancer efficacies. CONCLUSION: The prepared formulations could thus be considered as green anticancer agents in complementary medicine and therapies.

Protective effect of alamandine on doxorubicin­induced nephrotoxicity in rats.

BACKGROUND: This study aimed to evaluate the protective effects of alamandine, a new member of the angiotensin family, against doxorubicin (DOX)-induced nephrotoxicity in rats. METHODS: Rats were intraperitoneally injected with DOX (3.750 mg/kg/week) to reach a total cumulative dose of 15 mg/kg by day 35. Alamandine (50 µg/kg/day) was administered to the rats via mini-osmotic pumps for 42 days. At the end of the experiment, rats were placed in the metabolic cages for 24 h so that their water intake and urine output could be measured. After scarification, the rats' serum and kidney tissues were collected, and biochemical, histopathological, and immunohistochemical studies were carried out. RESULTS: DOX administration yielded increases in pro-inflammatory cytokines, including interleukin (IL)-1ß and IL-6, pro-fibrotic proteins transforming growth factor-ß (TGF-ß), pro-inflammatory transcription factor nuclear kappa B (NF-κB), kidney malondialdehyde (MDA), creatinine clearance, blood urea nitrogen (BUN), and water intake. On the other hand, the DOX-treated group exhibited decreased renal superoxide dismutase (SOD), renal glutathione peroxidase (GPx) activity, and urinary output. Alamandine co-therapy decreased these effects, as confirmed by histopathology and immunohistochemical analysis. CONCLUSIONS: The results suggest that alamandine can prevent nephrotoxicity induced by DOX in rats.

Alamandine significantly reduces doxorubicin-induced cardiotoxicity in rats.

Doxorubicin (DOX) is an anthracycline antibiotic. Despite its unwanted side effects, it has been successfully used in tumor therapy. Given that oxidative stress and inflammatory factors are essential to cardiotoxicity caused by DOX, we assumed that alamandine, which enhances endogenous antioxidants and has anti-inflammatory effects, may prevent DOX-induced cardiotoxicity. Rats received DOX (3.75 mg/kg) i.p on days 14, 21, 28, and 35 (total cumulative dose = 15 mg/kg) and alamandine (50 µg/kg/day) via mini-osmotic pumps for 42 days. At the end of the 42-day period, we evaluated hemodynamic parameters, electrocardiogram, cardiac troponin I (cTnI), superoxidase dismutase (SOD), total antioxidant capacity (TAC), malondialdehyde (MDA), inflammatory cytokines (tumor necrosis factor-α (TNF-α), IL-1ß, NF-κB), apoptosis markers (caspase 3), and histopathology of haemotoxylin- and eosin-stained cardiac muscle fibers were evaluated. DOX significantly increased QT, corrected QT (QTc), and RR intervals. Alamandine co-therapy prevented ECG changes. Alamandine administration restored DOX-induced disruptions in the cardiac muscle architecture and vascular congestion. Alamandine co-therapy also alleviated other effects of DOX, including cardiac contractility, decreased systolic and diastolic blood pressure, and increased left ventricular end-diastolic pressure. Moreover, alamandine co-therapy substantially decreased the elevation of oxidative stress markers, inflammatory cytokines, and caspase 3 in DOX-treated rats. The results suggest that alamandine reduced DOX-induced cardiotoxicity via antioxidant, anti-inflammatory, and anti-apoptotic activities.

Inhibitory Effect of Viola odorata Extract on Tumor Growth and Metastasis in 4T1 Breast Cancer Model.

Viola odorata as a medical herb is used in liver disorders and relieving cancer pain. In the present study, the cytotoxic, antioxidant, and anti-metastatic properties of Viola odorata hydro-alcoholic extract (VOE) were investigated in 4T1 breast cancer model. After treatment of 4T1 breast cancer cells with VOE, cell viability was measured by MTT assay. The implanted mice were treated with different concentration of VOE (50, 150 and 250 mg/kg) for 21 days. Levels of lactate dehydrogenase (LDH), γ -glutamyl transferase (GGT), alkaline phosphatase (ALP), carcinoembryonic antigen (CEA) and cancer antigen 15-3(CA15-3) in serum, and also catalase (CAT) and superoxide dismutase (SOD) activities in tumor tissue were measured. Metastatic rate was investigated in liver, spleen and lung tissues. VOE decreased cell viability of 4T1 cells, significantly. VOE significantly inhibited the cell proliferation, but not vasculature in the tumors that revealed by immunohistochemical analysis for Ki-67 and CD31 expression, respectively. VOE increased the Bax/Bcl-2 ratio in VOE250-treated group compared to control group. Serum analysis showed that treatment with 250 mg/kg of VOE significantly reduced LDH (not ALP and GGT) levels compared to controls. No linear correlation was found between the values of CEA and CA15-3 with tumor size. The rate of CAT activity was increased in VOE250-treated rats whereas, CAT and SOD activities were reduced in VOE50 group. VOE250 significantly decreased the metastatic rate in liver and lung compared to the other doses of VOE. Consequently, Viola odorata has cytotoxic effects on 4T1 cells and affects antioxidant activity and metastasis in breast cancer.

Mitochondrial DNA variation, genetic structure and demographic history of Iranian populations.

In order to survey the evolutionary history and impact of historical events on the genetic structure of Iranian people, the HV2 region of 141 mtDNA sequences related to six Iranian populations were analyzed. Slight and non-significant FST distances among the Central-western Persian speaking populations of Iran testify to the common origin of these populations from one proto-population. Mismatch distribution suggests that this proto-Iranian population started to colonize Iran about 30000 years ago which is almost consistent with the timing of arrival and colonization of western Asia by the anatomically modern human. Star-like haplotype network structures, significant and negative Tajima's D (D=-2.08, P<0.05) and unimodal mismatch distributions support the genetic effects of this expansion. Iranian populations presented mtDNA lineages that clearly belong to the European gene pool (i.e. H and U), while the Mashhad population was characterized by the presence of eastern and central Asian mtDNA lineages (i.e. M, B and D). Furthermore, the low diversity (h=0.428) observed in Mashhad may indicated the presence of inbreeding, drift or bottleneck events. The application of Monmonier's maximum differences algorithm revealed a geographic zone of genetic discontinuity between the Arab people of Khuzestan and rest of Iranian populations. Geographical factors, in cooperation with cultural/linguistic differences, are the main reasons for this differentiation. The lack of a sharp geographical or ethno-linguistic structure for mtDNA HV2 sequence diversity was statistically supported by AMOVA and Mantel (r=0.19, P<0.05) tests.

Effect of interaction between testosterone and morphine on serum ghrelin concentration in sheep fed on different dietary energy levels.

BACKGROUND: Ghrelin plays an important role in the regulation of food intake and body weight. It also decreases testosterone and opioid secretion. OBJECTIVES: The goal of the present study was to investigate the effect of testosterone, morphine or simultaneous injection of testosterone and morphine on mean serum ghrelin concentration in sheep. MATERIALS AND METHODS: Ten sheep were divided into two groups (n = 5 in each group), they were fed with either 50 % or 100 % of their dietary energy needs for 10 days. Body weight was measured on the 1st and 10th day of the experiment. Animals in both groups received testosterone (60 µg/kg), morphine (0.15 mg/kg), or a simultaneous infusion of testosterone (60 µg/kg) and morphine (0.15 mg/kg), on the 8th, 9th, or 10th day of the experiment respectively. Blood samples were collected before and 2 hours after the infusions. Ghrelin concentration was determined by RIA (radio immunoassay). RESULTS: In the 50 % group, ghrelin concentrations increased significantly on the 8th day of the experiment, compared to the 1st day (P < 0.05). While in the 100 % group, no significant change was observed. In both groups the animals' body weight did not increase significantly on the 10th day compared to the 1st day. Testosterone significantly increased ghrelin levels after injection compared to before infusion, in both groups (P < 0.05). Morphine increased ghrelin concentration in both groups, but this increase was not statistically significant. Simultaneous injection of testosterone and morphine together, significantly increased ghrelin concentration following injection compared to before infusion, in both groups (P < 0.05). CONCLUSIONS: There is a direct correlation between food restriction, testosterone and ghrelin concentration in ruminants. However, a simultaneous injection of testosterone and morphine did not exert an additive effect on ghrelin secretion.