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

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

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