Effects of selegiline, a monoamine oxidase B inhibitor, on differentiation of P19 embryonal carcinoma stem cells, into neuron-like cells

Selegiline, the irreversible inhibitor of monoamine oxidase B (MAO-B), is currently used to treat Parkinson's disease. However, the mechanism of action of selegiline is complex and cannot be explained solely by its MAO-B inhibitory action. It stimulates gene expression, as well as expression of a number of mRNAs or proteins in nerve and glial cells. Direct neuroprotective and antiapoptotic actions of selegiline have previously been observed in vitro. Previous studies showed that selegiline can induce neuronal phenotype in cultured bone marrow stem cells and embryonic stem cells. Embryonal carcinoma (EC) cells are developmentaly pluripotene cells which can be differentiated into all cell types under the appropriate conditions. The present study was carried out to examine the effects of selegiline on undifferentiated P19 EC cells. The results showed that selegiline treatment had a dramatic effect on neuronal morphology. It induced the differentiation of EC cells into neuron-like cells in a concentration-dependent manner. The peak response was in a dose of selegiline significantly lower than required for MAO-B inhibition. The differentiated cells were immunoreactive for neuron-specific proteins, synaptophysin, and beta-III tubulin. Stem cell therapy has been considered as an ideal option for the treatment of neurodegenerative diseases. Generation of neurons from stem cells could serve as a source for potential cell therapy. This study suggests the potential use of combined selegiline and stem cell therapy to improve deficits in neurodegenerative diseases

The Inhibitory Effects of Eucalyptus Extract on Herpes Simplex Virus-1 Replication in Baby Hamster Kidney Cells

Background and Objectives: In recent years, following the increasing of drug resistant strain of viruses, there has been an increasing interest in the use of natural substances with antiviral activity and low side effects. One of these herbal medicines, Eucaliptus, has shown some therapeutic effects including antibacterial and antiviral activities. Therefore, this study aimed to determine the minimum inhibitory concentration of hydroalchoholic extract of Eucaliptus on Herpes simplex virus-1 (HSV-1) in vitro. Methods: In this experimental study, the hydroalchoholic extract of Eucalyptus leaves was prepared using 70% ethanol through maceration method. Baby Hamster Kidney (BHK) cells were grown in monolayer culture with Dulbecco's modified Eagle's medium (DMEM) supplemented with 5% fetal calf serum and plated onto 48-well culture plates. 50% cytotoxic concentration (CC50%) of the extract on BHK cells was determined, and subsequently 50% inhibitory concentration (IC50%) of the herbal extract on replication of HSV-1 both in extracellular and intracellular cases was assessed. Results: Based on Probit analysis, CC50% of the extract was 0.650mg/ml. Significant relationships between the concentration of the extract and cell death in the cell studied were shown using the Probit model (p<0.01). IC50s of the extract on the virus before cellular attachment and after entering the cells were 456.82µg/ml and 180.75µg/ml, respectively. Based on the model, with increasing the extract concentration, the percentage of inhibition of cytopathic effect (CPE) in both stages was increased (p<0.05). Conclusion: Based on the findings of this study, hydroalchoholic extract of Eucaliptus could be probably an appropriate anti herpetic herbal medicine

The inhibitory effects of myrtle (Myrtus communis) extract on Herpes simplex virus-1 replication in Baby Hamster Kidney cells

زمینه و هدف: در سال های اخیر با افزایش سویه های مقاوم به دارو در انواع ویروس ها، یافتن مواد طبیعی با خواص ضد ویروسی که دارای اثرات جانبی کمتری نیز باشند مورد توجه محققین قرار گرفته است. گیاه مورد دارای اثرات مختلف درمانی از جمله اثر ضد باکتریایی و ضد قارچی می باشد. لذا این مطالعه با هدف تعیین حداقل غلظت بازدارندگی عصاره هیدروالکلی مورد روی ویروس هرپس سیمپلکس ویروس در کشت سلول صورت گرفته است. روش بررسی: در این مطالعه تجربی عصاره هیدروالکلی برگ های مورد به روش خیساندن با اتانول 70 تهیه شد. سلول(Baby Hamster Kidney) BHK در محیط کشت حاوی 5 سرم جنین گوساله در میکروپلیت های 48 خانه ای کشت گردید. پس از تعیین 50CC (Cytotoxic Concentration) عصاره بر روی سلول های BHK، اثر ممانعت کنندگی (50Inhibitory concentration= IC) آن بر تکثیر ویروس هرپس سیمپلکس نوع 1 (HSV1) در دومرحله جلوگیری از اتصال و تکثیر بعد از اتصال ارزیابی گردید. داده ها به کمک آزمون آماری پروبیت در هر مرحله محاسبه گردید. یافته ها: عصاره 96/4 میلی گرم بر میلی لیتر تعیین گردید. مدل پروبیت ارتباط معنی داری بین غلظت عصاره مورد و مرگ سلول ها نشان داد (001/0

Efficacy of Myrtus communis L. to inactivate the hydatid cyst protoscoleces

Purpose: The present study aims to investigate the scolicidal effects of Myrtus communis L. essential oil against protoscoleces of hydatid cysts and also its toxicity in mice model. Materials and Methods: Protoscoleces were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations of the essential oil (12.5–100 μl/ml) were used for 5–30 min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining). Moreover, 48 male NMRI mice were used to determine the acute and sub-acute toxicity of M. communis essential oil. One-way ANOVA with Tukey’s post-hoc test was used to assess differences between experimental groups. Results: Findings of the present study demonstrated that the M. communis essential oil at the concentration of 100 μl/ml after 5 min of exposure killed 100% protoscoleces. Similarly, the mean mortality rate of protoscoleces after 10 min of exposure to concentration of 50 μl/ml was 100%. However, lower concentrations (12.5 and 25 μl/ml) of M. communis essential oil provoked a delayed protoscolicidal effects. The LD50 values of intraperitoneal injection of the M. communis essential oil was 2.23 mL/kg body wt. No significant difference (p > .05) was observed in the clinical chemistry and hematological parameters following oral administrations of M. communis essential oil at the doses 0.05, 0.1, 0.2, and 0.4 mL/kg for 14 days. Conclusion: The results showed potent scolicidal activity of M. communis with no significant toxicity, which might be used as a natural scolicidal agent in hydatid cyst surgery

Evaluation of lentiviral vector-based green fluorescent protein expression in human gastric cancer cell line evaluation of expression lentivirus vector-based GFP in AGS

Background and aims : Human immunodeficiency virus type 1 (HIV-1) based-lentivirus vector is one of the most promising viral vectors for gene delivery in different cell lines including gastric cell lines. Therefore, the aim of this study was to produce a lentivirus vector for transduction and expression of green fluorescent protein (GFP) in human gastric cancer cell line, AGS. Materials and Methods: In this piece of work, Escherichia coli HB101 was transformed with plasmids psPAX2, pTD, and pMD2.G, following the purification of which their DNA was extracted along with their quantity and quality evaluated to be used in the next experiments. Subsequently, to produce the vector, the packaging cells were transfected with the plasmids and the vector containing supernatant was collected and purified using ultracentrifuge. ELISA was used to confirm the construction of the vector. Fluorescent microscopy and flow cytometry were used to check the expression of GFP in the cell line and to calculate the percentage of GFP expression, respectively. Results: In this study, the results of ELISA confirmed the construction of the plasmid used in this study. AGS cells were infected with viruses produced to detect the viral activity and GFP expression was evaluated by fluorescence microscopy and flow cytometry after 72 hours. Based on the results of flow cytometry, GFP was expressed in over 90% of transduced AGS cells. Conclusion: The results of this study showed that lentiviral vector is a highly efficient vector for expression of GFP gene in AGS cell line. Keywords: Lentivirus-based vector, Transfection, Transduction, GFP, AG

Efficacy of Myrtus communis L. to Inactivate the Hydatid Cyst Protoscoleces

Purpose: The present study aims to investigate the scolicidal effects of Myrtus communis L. essential oil against protoscoleces of hydatid cysts and also its toxicity in mice model. Materials and Methods: Protoscoleces were aseptically aspirated from sheep livers having hydatid cysts. Various concentrations of the essential oil (12.5–100 μl/ml) were used for 5–30 min. Viability of protoscoleces was confirmed using eosin exclusion test (0.1% eosin staining). Moreover, 48 male NMRI mice were used to determine the acute and sub-acute toxicity of M. communis essential oil. One-way ANOVA with Tukey’s post-hoc test was used to assess differences between experimental groups. Results: Findings of the present study demonstrated that the M. communis essential oil at the concentration of 100 μl/ml after 5 min of exposure killed 100% protoscoleces. Similarly, the mean mortality rate of protoscoleces after 10 min of exposure to concentration of 50 μl/ml was 100%. However, lower concentrations (12.5 and 25 μl/ml) of M. communis essential oil provoked a delayed protoscolicidal effects. The LD50 values of intraperitoneal injection of the M. communis essential oil was 2.23 mL/kg body wt. No significant difference (p > .05) was observed in the clinical chemistry and hematological parameters following oral administrations of M. communis essential oil at the doses 0.05, 0.1, 0.2, and 0.4 mL/kg for 14 days. Conclusion: The results showed potent scolicidal activity of M. communis with no significant toxicity, which might be used as a natural scolicidal agent in hydatid cyst surgery

Screening of three common mtDNA mutations among subjects with autosomal recessive non-syndromic hearing loss in Sistan va Baluchestan province, Iran

Background: Non-syndromic hearing loss may be induced by mutations in both nuclear and mitochondrial genes. Mutations in mtDNA are present in less than 1% of the children with pre-lingual deafness but are more prevalent later. Most of the molecular defects responsible for mitochondrial disorder, associated with hearing loss may be induced by mutations in the 12SrRNA and tRNA genes. This aim of this study was to investigate the frequency of three common mtDNA mutations including A1555G, A3243G and A7445G in a cohort of autosomal recessive non-syndromic hearing loss (ARNSHL) subjects in Sistan va Baluchestan province. Material and Methods: In this descriptive- experimental based study, a total of 110. ARNSHL subjects from Sistan va Baluchestan province were investigated for three common mtDNA mutations using PCR-RFLP procedure. The possible mutations were confirmed by direct sequencing. Results: None of the A1555G and A7445G mutations were detected in this study. However, we found one sample to carry A3243G mutation (0.9%). Moreover abolishing a MTTL1 restriction site close to A3243G mutation revealed a G3316A allelic variant in 0.9% of patients studied. Conclusion: This study showed that mtDNA mutations are responsible for less than 1% of pre-lingual ARNSHL associated subjects. The present study will improve the genetic counseling of hearing impaired patients in Sistan va Baluchestan province, Iran

Astaxanthin ameliorates inflammation, oxidative stress, and reproductive outcomes in endometriosis patients undergoing assisted reproduction: A randomized, triple-blind placebo-controlled clinical trial

PurposeIn a randomized, triple-blind, placebo-controlled clinical trial (RCT) including 50 infertile women with endometriosis candidate for assisted reproductive techniques (ART), we studied the effect of Astaxanthin (AST) on pro-inflammatory cytokines, oxidative stress (OS) markers, and early pregnancy outcomes.MethodsBefore and after 12 weeks of AST treatment (6 mg per day), blood serum and follicular fluid (FF) samples were collected from 50 infertile women with endometriosis stage III/IV undergoing ART. Pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) and OS markers (malondialdehyde [MDA], superoxide dismutase [SOD], catalase [CAT], and total antioxidant capacity [TAC]) were measured in the serum and FF. ART outcomes were also compared between the groups.ResultsIncreased serum levels of TAC (398.661 ± 57.686 vs. 364.746 ± 51.569; P = 0.004) and SOD (13.458 ± 7.276 vs. 9.040 ± 5.155; P = 0.010) were observed after AST therapy in the treatment group. Furthermore, serum MDA (14.619 ± 2.505 vs. 15.939 ± 1.512; P = 0.031) decreased significantly following antioxidant treatment. In addition, significantly lower serum levels of IL-1β (4.515 ± 0.907 vs. 6.8760 ± 0.8478; P = 0.000), IL-6 (5.516 ± 0.646 vs. 5.0543 ± 0.709; P = 0.024) and TNF-α (2.520 ± 0.525 vs. 2.968 ± 0.548; P = 0.038) were observed after AST treatment. In addition, AST supplementation led to an improved number of oocytes retrieved (14.60 ± 7.79 vs. 9.84 ± 6.44; P = 0.043), number of mature (MII) oocytes (10.48 ± 6.665 vs. 6.72 ± 4.3; P = 0.041), and high-quality embryos (4.52 ± 2.41 vs. 2.72 ± 2.40; P = 0.024).ConclusionAST pretreatment can modulate inflammation and OS in endometriosis-induced infertile patients. ART outcomes also improved after 12 weeks of AST therapy. Our results suggest that AST can be a potential therapeutic target for infertile patients with endometriosis undergoing ART

Effect of Screw Access Channel Filling Method and Cement Type on Retention of Implant-Supported Fixed Restorations

Introduction: There is limited data on the factors affecting the retention of cemented fixed prostheses to implant abutment. The aim of this study was to evaluate the effect of screw access channel filling method and cement type on retention of implant-supported fixed restorations.  Materials and Methods: In this experimental study, 40 implant analogs were mounted in autopolymerizing acrylic resin blocks, and two-piece titanium abutments were placed in each implant analog. Twenty abutment samples were completely filled with silicone, and 20 other samples were filled partially. In each of the study groups, Temp Bond® eugenol-containing temporary cement was used for 10 samples, while in another 10 samples non-eugenol temporary cements were utilized. Prior to the retention test, samples were placed in the rmocycling machine with 1000 cycles for 24 h. Each sample was stretched using a Universal Pull-out Test Machine with a force of 5000 N. The required load for removing the crown was recorded. The data was analyzed USING two-way ANOVA and least square difference (α=0.05). Results: Among the four groups, the highest retention rate was observed in the group of partial screw access channel filling with eugenol cement. Also, the rate of retention in the group of complete screw access channel filling with non-eugenol cement was significantly lower than in any other group. A significant difference was observed between all the groups except for the groups of complete screw access channel filling with eugenol cement and partial screw access channel filling with non-eugenol cement (P=0.27). Conclusion: The mean rate of retention in partial access cavity filling group was greater than that of the complete access cavity filling group; moreover, this rate was higher in the eugenol cement group than the non-eugenol cement group