Gene selection using pyramid gravitational search algorithm.

Genetics play a prominent role in the development and progression of malignant neoplasms. Identification of the relevant genes is a high-dimensional data processing problem. Pyramid gravitational search algorithm (PGSA), a hybrid method in which the number of genes is cyclically reduced is proposed to conquer the curse of dimensionality. PGSA consists of two elements, a filter and a wrapper method (inspired by the gravitational search algorithm) which iterates through cycles. The genes selected in each cycle are passed on to the subsequent cycles to further reduce the dimension. PGSA tries to maximize the classification accuracy using the most informative genes while reducing the number of genes. Results are reported on a multi-class microarray gene expression dataset for breast cancer. Several feature selection algorithms have been implemented to have a fair comparison. The PGSA ranked first in terms of accuracy (84.5%) with 73 genes. To check if the selected genes are meaningful in terms of patient's survival and response to therapy, protein-protein interaction network analysis has been applied on the genes. An interesting pattern was emerged when examining the genetic network. HSP90AA1, PTK2 and SRC genes were amongst the top-rated bottleneck genes, and DNA damage, cell adhesion and migration pathways are highly enriched in the network.

Metformin is a Novel Suppressor for Vimentin in Human Gastric Cancer Cell Line.

Vimentin, an intermediate filament of mesenchymal cells, is upregulated in epithelial-mesenchymal transition (EMT) and has a main role in cancer metastasis. As a new strategy to control metastatic outgrowth, EMT markers are generally inhibited using some drugs or specific siRNA. In this study, AGS gastric cancer cells were treated with metformin and vimentin-specific siRNA (vim-siRNA) for 48 h. The impact of metformin and vim-siRNA on vimentin downregulation in AGS cells were analyzed by quantitative PCR and Western blot. Following treatment with metformin and vim-siRNA, cell motility, migration and invasion abilities of AGS cells were also analyzed. The results showed that inhibition of vimentin due to metformin was comparable with the vim-siRNA. Furthermore, wound-healing and invasion assays showed a significant decrease in migration and invasion of AGS cells following metformin and vim-siRNA treatment. Our finding for the first time indicated that metformin can be an alternative to specific siRNA for inhibition of vimentin expression and migration of AGS cell line. Taken together, our data indicates that the use of metformin might have a priority to siRNA for inhibition of gastric cancer cell behaviors siRNA is more unstable and expensive than metformin, and needs special vehicles and delivery strategies for efficient transfection of cells. Further in vivo studies can reveal metformin's potential in inhibition of EMT and metastasis of cancer cells.

Effects of hydrogen peroxide, doxorubicin and ultraviolet irradiation on senescence of human dental pulp stem cells.

OBJECTIVE: The objective of this study was to evaluate the ability of three distinct agents on the induction of senescence in human dental pulp stem cells (DPSCs). DESIGN: DPSCs from three separate donors were treated with H2O2, doxorubicin and ultraviolet (UV) irradiation. The response of the cells to the three agents was assayed by specific staining for SA-ßGal, RT-qPCR and flow cytometry. RESULTS: The results showed that incubation with 100 µM H2O2 and 20 nM Doxorubicin for seven days led to senescence in all donors' cells equally. Interestingly, UV irradiation for just one minute was sufficient to induce senescence in the cells. The SA-ßGal positive senescent cells were arrested in G1 phase and their S phase was significantly reduced as analyzed by flow cytometry. Significant increment in p21 and BTG1 expression and decrement in CCND1 expression also confirmed the cells have been arrested and get senescent via p53-p21 pathway. CONCLUSION: All three agents successfully triggered senescence in the cells. There was no significant difference in the capacity of the three donor's cells for senescence. To avoid premature senescence in stem cell in vitro, it is recommended to avoid unnecessary exposure of the cell to fluorescent and UV light. Moreover, to prevent ROS production, we recommend using a separate incubator with low oxygen content for cell culture, if possible.

Cytotoxic and anti-proliferative effects of Rosa beggeriana Schrenk extracts on human liver and breast cancer cells.

OBJECTIVE: Rosa beggeriana Schrenk has been consumed in Iranian traditional medicine. In contrary to its close species (e.g. R. canina), there is no data on its medicinal properties. Therefore, we explored possible cytotoxic effects of R. beggeriana against two cancer cell lines. MATERIALS AND METHODS: The cytotoxic and anti-proliferative effects of R. beggeriana ethanolic and aqueous extracts on human liver cancer cells (LCLPI 11), breast cancer cells (MCF-7) and fibroblast-like cells (HSkMC) were evaluated by MTT, BrdU and TUNEL assays. RESULTS: Following 48 h, IC50 values for LCL-PI11 and MCF-7 cells were found to be 3.9 and 4.2 µg/mL for aqueous extract, and 2.3 and 2.7 µg/mL for ethanolic extract, respectively.BrdU assay data verified the MTT results and showed that both extracts inhibit cell proliferation as much as 5-fluorouracil does (p<0.05). The ethanolic extract had a more marked inhibitory effect compared to the aqueous extract (p<0.05). Besides both extracts were less effective against HSKMC cells compared to other cells lines.TUNEL assay results demonstrated that following 48 h, the aqueous extract induced about 19 and 24% apoptotic death in the LCL-PI 11 and MCF-7 cells, respectively (p<0.05). While at the same time, the ethanolic extract was more potent and caused about 83 and 91% death in the LCL-PI 11 and MCF-7 cells, respectively (p<0.05). CONCLUSION: These data indicate that both extracts have anti-proliferative and pro-apoptotic activities on these two cancer cell lines and these effects were more pronounced then their activities against normal cells. Also, the ethanolic extract was more potent than the aqueous extract. Further researches are necessary for finding and isolating effective anticancer ingredient of R. beggeriana.

Controlled release of lawsone from polycaprolactone/gelatin electrospun nano fibers for skin tissue regeneration.

In this study, we aimed to evaluate the role of electrospun nanofibers containing lawsone (2-hydroxy-1,4-naphthoquinone) in wound healing. Different concentrations of lawsone (0.5%, 1%, 1.5%) were electrospun in polycaprolactone-gelatin (PCL/Gel) polymers in core-shell architecture. Nanofibers were characterized using scanning electron microscopy (SEM), Transmission electron microscopy (TEM). Coaxial electrospinning of PCL/Gel/lawsone (PCL/Gel/Law) scaffolds prolonged the release of lawsone over a period of 20 days. In vitro bioactivity of fibers on human gingiva fibroblast cells (HGF) was evaluated by MTT assay. The PCL/Gel/Law 1% increased cell attachment and proliferation significantly. Additionally, the in vitro gene expression of transforming growth factor ß (TGF-B1), collagen (COL1) and epidermal growth factor (EGF) was monitored using RT-qPCR technique, which there was significant increase in TGF-B1 and COL1 gene expression in PCL/Gel/Law 0.5% and 1% mats. In vivo wound healing activity of lawsone loaded scaffolds in rat wound model revealed that the PCL/Gel/Law 1% had the highest impact on healing by increasing re-epithelialization of the wound after 14 days. It can be inferred that lawsone 1% incorporation in the core of PCL/Gel electrospun nanofibers has excellent characteristics and can be used as wound dressing patch in medicine.

In-Vitro Anti-Proliferative and Pro-Apoptotic Properties of Sutureja Khuzestanica on Human Breast Cancer Cell Line (MCF-7) and Its Synergic Effects with Anticancer Drug Vincristine.

Satureja khuzestanica Jamzad (Marzeh Khuzestani in Persian) is an endemic plant that is widely distributed in the southern part of Iran. Despite the number of papers published on this plant, no one has focused on its anticancer effects. Therefore, the present study was designed to investigate the selective cytotoxic and anti-proliferative properties of satureja khuzestanica total extract (SKE). MCF-7 human breast cancer cells were used in this study. Cytotoxicity of the extract was determined using MTT and neutral red assaysafter 24 h treatment period. Biochemical markers of apoptosis (caspase-3, Bax, Bcl-2) and cell proliferation (cyclin D1) were evaluated by immunoblotting. Vincristine was used to evaluate the synergic effect of extract with an anticancer drug. The data showed that treatment of cells with SKE (150 and 200 µg/mL for 24 h) significantly reduced cell viability, activated caspase 3 and increased Bax/Bcl2 ratio. In addition, cyclin D1 expression was significantly decreased in the SKE-treated cells. In addition, concomitant treatment of the MCF-7 cells with SKE and vincristine produced a potent anti-proliferative and pro-apoptotic effect compared to extract or drug alone. In conclusion, satureja extract has a potential anti-cancer property against human breast cancer cells and its combination with chemotherapeutic agent vincristine may induce cell death effectively and be a potent modality to treat this type of cancer.

Long noncoding RNA VIM-AS1 promotes colorectal cancer progression and metastasis by inducing EMT.

Emerging evidence indicates that lncRNAs play crucial roles in the initiation and progression of various malignant tumors. VIM-AS1 RNA is an lncRNA that transcribes from a shared bidirectional promoter with vimentin mRNA and its function in cancer cells is largely unknown. This study assessed the clinical significance of VIM-AS1 expression in colorectal cancer (CRC). We found that the VIM-AS1 transcript was significantly upregulated in high-grade, lymph node metastasis and vascular invasion tumors. Loss-of-function experiments revealed that the downregulation of VIM-AS1 could inhibit tumor cell proliferation by inducing apoptosis, cellular senescence and arresting the cell cycle. Moreover, the obtained data demonstrated that VIM-AS1 might play a crucial role in cell migration as well as the epithelial to mesenchymal transition (EMT) of CRC cells. Collectively, for the first time, our data provide novel evidence for the biological and clinical significance of VIM-AS1 expression in CRC. Further, the findings of this study suggest that VIM-AS1 promotes tumor growth and metastasis by inducing EMT in CRC cells and could be considered as a novel tumor marker with probable value in diagnosis and CRC treatment.

Cytotoxic effects of Fritillaria imperialis L. extracts on human liver cancer cells, breast cancer cells and fibroblast-like cells.

In this study, we evaluated the cytotoxic effects of Fritillaria imperialis L. aqueous and ethanolic extracts on human liver cancer cells (LCL-PI 11), breast adenocarcinoma cells (MCF-7) and fibroblast-like cells (HSkMC). MTT, BrdU and TUNEL assays were used to detect cytotoxicity, proliferation rate and apoptotic death. Following 48h, the IC50 values for LCL-PI11 and MCF-7 cells were 4.2 and 3.9µg/mL of aqueous extract, and 1.7 and 1.3µg/mL of ethanolic extract, respectively, which was comparable to that of 5-FU. BrdU assay data verified that both extracts inhibited cell proliferation more preferentially on the two cancer cells. Exposure of LCL-PI 11 and MCF-7 cells to 2.4µg/mL of aqueous extract for 24h resulted in 29% and 32% apoptotic death. Surprisingly the ethanolic extract killed nearly all of the cells at 1.6µg/mL concentration. Collectively, our data indicated that both extracts have cytotoxic, cytostatic and pro-apoptotic activities against the two cancer cell lines. The ethanolic extract was more potent than the aqueous extract and the fibroblast cell was found to be less influenced than both cancer cell lines. Further researches are necessary for chemical characterization and in vivo evaluation especially in animal models to identify the effective anticancer ingredient of F. imperialis.

Metformin inhibits gastric cancer cells metastatic traits through suppression of epithelial-mesenchymal transition in a glucose-independent manner.

Epithelial-mesenchymal transition (EMT), which is mainly recognized by upregulation of mesenchymal markers and movement of cells, is a critical stage occurred during embryo development and spreading cancerous cells. Metformin is an antidiabetic drug used in treatment of type 2 diabetes. EMT inhibitory effect of metformin has been studied in several cancers; however, it remains unknown in gastric cancer. The aim of the present study was to investigate the metformin effects on inhibition of EMT-related genes as well as migration and invasion of AGS gastric cancer cell line. Moreover, to study the effect of glucose on metformin-mediated EMT inhibition, all experiments were performed in two glucose levels, similar to non-fasting blood sugar (7.8 mM) and hyperglycemic (17.5 mM) conditions. The results showed reduction of mesenchymal markers, including vimentin and ß-catenin, and induction of epithelial marker, E-cadherin, by metformin in both glucose concentrations. Furthermore, wound-healing and invasion assays showed a significant decrease in cell migration and invasion after metformin treatment in both glucose levels. In conclusion, our results indicated that metformin strongly inhibited EMT of gastric cancer cells in conditions mimicking normo and hyperglycemic blood sugar.

Differential Expression of Hox and Notch Genes in Larval and Adult Stages of Echinococcus granulosus.

This investigation aimed to evaluate the differential expression of HoxB7 and notch genes in different developmental stages of Echinococcus granulosus sensu stricto. The expression of HoxB7 gene was observed at all developmental stages. Nevertheless, significant fold differences in the expression level was documented in the juvenile worm with 3 or more proglottids, the germinal layer from infected sheep, and the adult worm from an experimentally infected dog. The notch gene was expressed at all developmental stages of E. granulosus; however, the fold difference was significantly increased at the microcysts in monophasic culture medium and the germinal layer of infected sheep in comparison with other stages. The findings demonstrated that the 2 aforementioned genes evaluated in the present study were differentially expressed at different developmental stages of the parasite and may contribute to some important biological processes of E. granulosus.

Trimethylamine-N-Oxide Treatment Induces Changes in the ATP-Binding Cassette Transporter A1 and Scavenger Receptor A1 in Murine Macrophage J774A.1 cells.

BACKGROUND: Recently, trimethylamine N-oxide was introduced as a risk factor for atherosclerosis in terms of helping foam cell formation and worsening atherosclerosis complications. The present study was performed to investigate whether/how trimethylamine N-oxide is involved in regulation of ATP-binding cassette transporter A1 and scavenger receptor A1 in macrophages at both mRNA and protein levels. METHODS: Murine macrophage J774A.1 cells were treated with micromolar concentrations of trimethylamine N-oxide and 4-phenylbutyric acid, a chemical chaperon, for different time intervals. Tunicamycin was also used as a control for induction of endoplasmic reticulum stress. RESULTS: Similar to tunicamycin, trimethylamine N-oxide increased scavenger receptor A1 in all treatment periods, whereas ATP-binding cassette transporter A1 was only reduced 24 h post-treatment with trimethylamine N-oxide at both mRNA and protein levels. In contrast, 4-phenylbutyric acid failed to induce such changes in either scavenger receptor A1 or ATP-binding cassette transporter A1. CONCLUSIONS: The results of this study, in agreement with previous studies, confirm the mechanistic role of trimethylamine N-oxide in the upregulation of scavenger receptor A1, which potentially can promote its proatherogenic role. The results also showed downregulation of ATP-binding cassette transporter A1 in trimethylamine N-oxide treated macrophages which may indicate another possible proatherosclerotic mechanism for foam cell formation.

Isolation and evaluation of dental pulp stem cells from teeth with advanced periodontal disease.

INTRODUCTION: Successful isolation of mesenchymal stem cells from waste tissues might be extremely promising for developing stem cell-based therapies. This study aimed to explore whether cells retrieved from teeth extracted due to advanced periodontal disease present mesenchymal stem cell-like properties. METHODS: Pulp cells were isolated from 15 intact molars and 15 teeth with advanced periodontal disease. Cell proliferation and markers of mesenchymal stem cells were evaluated. RESULTS: Based on the RT-PCR and agarose gel electrophoresis, nucleostemin, Oct-4 and jmj2c, but not Nanog, were expressed in undifferentiated mesenchymal stem cells of both groups. Interestingly, diseased pulp exhibited higher gene expressions although it was not statistically significant. The average percentage of BrdU positive cells in the diseased group (84.4%, n = 5) was significantly higher than that of the control group (65.4%, n = 5) (t-test, P = 0.001). CONCLUSION: Our results indicate the successful isolation of mesenchymal stem cells from the pulp tissue of hopeless periodontally involved teeth.

A modified efficient method for dental pulp stem cell isolation.

BACKGROUND: Dental pulp stem cells can be used in regenerative endodontic therapy. The aim of this study was to introduce an efficient method for dental pulp stem cells isolation. MATERIALS AND METHODS: In this in-vitro study, 60 extracted human third molars were split and pulp tissue was extracted. Dental pulp stem cells were isolated by the following three different methods: (1) digestion of pulp by collagenase/dispase enzyme and culture of the released cells; (2) outgrowth of the cells by culture of undigested pulp pieces; (3) digestion of pulp tissue pieces and fixing them. The cells were cultured in minimum essential medium alpha modification (αMEM) medium supplemented with 20% fetal bovine serum(FBS) in humid 37°C incubator with 5% CO 2. The markers of stem cells were studied by reverse transcriptase polymerase chain reaction (PCR). The student t-test was used for comparing the means of independent groups. P <0.05 was considered as significant. RESULTS: The results indicated that by the first method a few cell colonies with homogenous morphology were detectable after 4 days, while in the outgrowth method more time was needed (10-12 days) to allow sufficient numbers of heterogeneous phenotype stem cells to migrate out of tissue. Interestingly, with the improved third method, we obtained stem cells successfully with about 60% efficiency after 2 days. The results of RT-PCR suggested the expression of Nanog, Oct-4, and Nucleostemin markers in the isolated cells from dental pulps. CONCLUSION: This study proposes a new method with high efficacy to obtain dental pulp stem cells in a short time.

Proapoptotic and Antiproliferative Effects of Thymus caramanicus on Human Breast Cancer Cell Line (MCF-7) and Its Interaction with Anticancer Drug Vincristine.

Thymus caramanicus Jalas is one of the species of thymus that grows in the wild in different regions of Iran. Traditionally, leaves of this plant are used in the treatment of diabetes, arthritis, and cancerous situation. Therefore, the present study was designed to investigate the selective cytotoxic and antiproliferative properties of Thymus caramanicus extract (TCE). MCF-7 human breast cancer cells were used in this study. Cytotoxicity of the extract was determined using MTT and neutral red assays. Biochemical markers of apoptosis (caspase 3, Bax, and Bcl-2) and cell proliferation (cyclin D1) were evaluated by immunoblotting. Vincristine was used as anticancer control drug in extract combination therapy. The data showed that incubation of cells with TCE (200 and 250 µ g/mL) significantly increased cell damage, activated caspase 3 and Bax/Bcl2 ratio. In addition, cyclin D1 was significantly decreased in TCE-treated cells. Furthermore, concomitant treatment of cells with extract and anticancer drug produced a significant cytotoxic effect as compared to extract or drugs alone. In conclusion, thymus extract has a potential proapoptotic/antiproliferative property against human breast cancer cells and its combination with chemotherapeutic agent vincristine may induce cell death effectively and be a potent modality to treat this type of cancer.

Isolation of copper oxide (CuO) nanoparticles resistant Pseudomonas strains from soil and investigation on possible mechanism for resistance.

The present study deals with isolation and characterization of copper oxide nanoparticles resistant Pseudomonas strains that were isolated from the soil collected from mining and refining sites of Sarcheshmeh copper mine in the Kerman Province of Iran. The three isolates were selected based on high level of copper oxide nanoparticles (CuO NPs) resistance. The isolates were authentically identified as Pseudomonas fluorescens CuO-1, Pseudomonas fluorescens CuO-2 and Pseudomonas sp. CuO-3 by morphological, biochemical and 16S rDNA gene sequencing analysis. The growth pattern of these isolates with all the studied CuO NPs concentrations was similar to that of control (without CuO NPs) indicating that CuO NPs would not affect the growth of isolated strains. A reduction in the amount of exopolysaccharides was observed after CuO NPs-P. fluorescens CuO-1 culture supernatant interaction. The Fourier transform infrared spectroscopy (FT-IR) peaks for the exopolysaccharides extracted from the bacterial culture supernatant and the interacted CuO NPs were almost similar. The exopolysaccharide capping of the CuO NPs was confirmed by FT-IR and X-ray diffraction analysis. The study of bacterial exopolysaccharides capped CuO NPs with E. coli PTCC 1338 and S. aureus PTCC 1113 showed less toxicity compared to uncoated CuO NPs. Our study suggests that the capping of nanoparticles by bacterially produced exopolysaccharides serve as the probable mechanism of tolerance.

Human Dental Pulp Stem Cells Differentiate into Oligodendrocyte Progenitors Using the Expression of Olig2 Transcription Factor.

The helix-loop-helix transcription factor Olig2 is essential for lineage determination of oligodendrocytes. Differentiation of stem cells into oligodendrocytes and transplanting them is a novel strategy for the repair of different demyelination diseases. Dental pulp stem cells (DPSCs) are of great interest in regenerative medicine due to their potential for repairing damaged tissues. In this study, DPSCs were isolated from human third molars and transfected with the human Olig2 gene as a differentiation inducer for the oligodendrogenic pathway. Following the differentiation procedure, the expression of Sox2, NG2, PDGFRα, Nestin, MBP, Olig2, Oct4, glial fibrillary acidic protein and A2B5 as stage-specific markers was studied by real-time RT-qPCR, immunocytochemistry and Western blot analysis. The cells were transplanted into a mouse model of local sciatic damage by lysolecithin as a model for demyelination. Oligodendrocyte progenitor cells (OPCs) actively remyelinated and recovered the lysolecithin-induced damages in the sciatic nerve as revealed by treadmill exercise, the von Frey filament test and hind paw withdrawal in response to a thermal stimulus. Recovery of behavioral reflexes occurred 2-6 weeks after OPC transplantation. The results demonstrate that the expression of Olig2 in DPSCs reduces the expression of stem cell markers and induces the development of oligodendrocyte progenitors as revealed by the emergence of oligodendrocyte markers. DPSCs could be programmed into oligodendrocyte progenitors and considered as a simple and valuable source for the cell therapy of neurodegenerative diseases.

Inhibition of 6-hydroxydopamine-induced PC12 cell apoptosis by olive (Olea europaea L.) leaf extract is performed by its main component oleuropein.

Parkinson disease (PD) is the most common progressive neurodegenerative disorder characterized by progressive death of midbrain dopaminergic neurons. Most neurodegenerative disease treatments are, at present, palliative. However, some natural herbal products have been shown to rescue neurons from death and apoptosis in some of neurodegenerative diseases. Not only Olea europaea L. olive oil, but also the leaves of this plant have been used for medical purposes. Olive leaf extract (OLE) is being used by people as a drink across the world and as an integral ingredient in their desire to maintain and improve their health. Here, we investigated the effects of OLE and its main phenolic component oleuropein on 6-hydroxydopamine (6-OHDA)-induced toxicity in rat adrenal pheochromocytoma (PC12) cells as an in vitro model of PD. Cell damage was induced by 150 µM 6-OHDA. The cell survival rate was examined by MTT assay. Generation of intra-cellular reactive oxygen species (ROS) was studied using fluorescence spectrophotometry. Immunoblotting and DNA analysis were also employed to determine the levels of biochemical markers of apoptosis in the cells. The data showed that 6-OHDA could decrease the viability of the cells. In addition, intra-cellular ROS, activated caspase 3, Bax/Bcl-2 ratio, as well as DNA fragmentation were significantly increased in 6-OHDA-treated cells. Incubation of cells with OLE (400 and 600 µg/mL) and oleuropein (20 and 25 µg/mL) could decrease cell damage and reduce biochemical markers of cell death. The results suggest that OLE and oleuropein have anti-oxidant protective effects against 6-OHDA-induced PC12 cell damage. The protective effects of OLE and oleuropein are correlative with their anti-oxidative and anti-apoptotic properties and suggest their therapeutic potential in the treatment of PD.

Mutation Analysis of TP53 Tumor Suppressor Gene in Colorectal Cancer in Patients from Iran (Kerman Province).

OBJECTIVES: P53 is an important tumor suppressor, which is mutated in later stages of many cancers and leads to resistance to chemotherapy. The aim of this study was to reveal mutations of TP53 in colorectal cancer in Kerman province. MATERIALS AND METHODS: A total of Forty-three colon cancer specimens as paraffin block or fresh tissues, which passed stage IIIA, were selected. Three exons 5, 7 and 8 of P53 were amplified by PCR technique and sequenced directly. RESULTS: The results showed two deletions at codon 140 and 142 in one tumor sample. GAT→AAT mutation at codon 184, and CGG→TGG mutation at codon 248 were seen in some tumor samples. Some mutations were also observed in middle of intron 7 in some tumor or normal tissues. CONCLUSION: Some of those patients with mutation in P53 gene had metastasis in other organs. Therefore, genetic test before chemotherapy is helpful for successful treatment.

Expression analysis and purification of human recombinant tissue type plasminogen activator (rt-PA) from transgenic tobacco plants.

Recombinant tissue-type plasminogen activator (rt-PA) has been produced in different hosts. In this research, transgenic tobacco was selected for production of human rt-PA. Transgenic plants were analyzed by polymerase chain reaction (PCR) and reverse-transcription (RT)-PCR. The protein was extracted by Lysine Sepharose chromatography column and was further purified by HiTrap desalting column. The function of eluted protein was analyzed on zymography gel. The results showed that the 1.7-kb cDNA of tissue-type plasminogen activator (t-PA) (as well as a shortened 650-bp transcript of t-PA) has been expressed in transgenic plants. The anticipated 63-kD protein band and an additional 53-kD protein were observed in transgenic plants. Finally, zymography assay revealed that the purified rt-PA has anticipated appropriate activity comparable to a positive control drug (Alteplase). On the whole, we can say that transgenic tobacco is a good alternative host for production of t-PA.

NGF and BDNF expression drop off in neurally differentiated bone marrow stromal stem cells.

Bone marrow stromal stem cells (BMSC) express two neurotrophins nerve growth factor (NGF) and brain derived growth factor (BDNF) constitutively and can be differentiated into neuronal-like cells and used to treat neural injuries and diseases. The neurotrophins are required for repair of neural tissues. However, it is not evident whether these cells supply the sufficient amounts of the functional growth factors following neuronal differentiation. This study investigates the expression of NGF, BDNF and their processing enzymes Prohormone convertases (PC) Furin, PC5 and PC6 by Real-time RT-PCR during neural differentiation of rat BMSC. The results showed that all inspected processing enzymes are expressed in the cells. The expression of NGF, BDNF and PC5 decreases following differentiation. In addition, BMSCs express Survivin, an anti-apoptotic gene; however, the differentiated cells reduce its expression similar to two neurotrophins, which could make them susceptible to apoptotic death.