Effects of inoculation with four mycorrhizal species on seed phenolic and fatty acids of sesame plants grown under different irrigation regimes.

This study evaluated the interaction effects of irrigation level (well-watered and water stress conditions) and inoculation by different mycorrhizal species (non-inoculated, Funneliformis mosseae, Rhizophagus irregularis, Claroideoglomus claroideum, and Glomus fasciculatum) on mycorrhizal colonization, antioxidant activity, seed yield and oil quality of two sesame cultivars (Yekta and Naz). Water deficit decreased mycorrhizal colonization, seed yield and oil concentration but increased antioxidant activity and seed total phenol and flavonoid concentrations. However, mycorrhizal inoculation increased antioxidant activity, seed yield, oil concentration and total phenolic and flavonoids. The lowest reduction by water stress and the highest increase by inoculation in seed yield were observed in Naz plants inoculated by Cl. claroideum. Principal component analysis showed the highest differentiation effect of water stress compared to mycorrhizal inoculation on both cultivars, indicating the relative sensitivity of the two cultivars to water deficit. However, the application of different species of mycorrhizal fungi versus the non-inoculation conditions was somewhat discriminative. In terms of fatty acids, in most cases, water stress increased oleic, palmitic and stearic acids and decreased linoleic and linolenic acids but inoculation increased oleic and linoleic acids and decreased linolenic, palmitic and stearic acids. Regarding phenolic and flavonoids components, the contents of chlorogenic and caffeic acids were increased by water stress but no consistent trend was noted in response to water stress for the other compounds. Mycorrhizal inoculation generally decreased chlorogenic acid but increased gallic, caffeic, p-coumaric, and ferulic acids. In conclusion, the results of the present study may help to increase the level of valuable compounds in sesame for further pharmaceutical purposes under water stress conditions and mycorrhizal symbiosis.

Downregulation of Stemness Genes and Induction of Necrosis in Rat LA7 Cancer Stem Cells Induced Tumors Treated with Starved Fibroblasts Culture Supernatant.

BACKGROUND: Stem cell differentiation therapy is a promising strategy in cancer treatment. we show that protein cocktail prepared from serum starved fibroblasts has therapeutic potential based on this strategy. METHODS: The condition medium was prepared from foreskin isolated fibroblasts and analyzed by Liquid chromatography electrospray ionization mass spectrometry-mass spectrometry (LC-ESI-MS/MS). LA7 mammary gland cancer stem cells originated tumors were induced in Sprague Dawley rats. The rats treated subcutaneously with DMEM (group A), condition medium (group B), or normal saline (group C) once daily for 7 days. Then the tumors were removed and divided into the two parts, one part was used to quantify gene expression by stem-loop RT-qPCR assay and the other part was used for Hematoxylin & Eosin (H & E), Giemsa, and immunohistochemistry (IHC) staining. RESULTS: All induced tumors appeared as sarcomatoid carcinoma (SC). Immunohistochemistry staining confirmed this conclusion by recognizing the tumor as Ki67+, cytokeratin+, vimentine+, and estrogen receptor negative SC. RT-qPCR analysis revealed that Oct4-, Sox-2, Nanog- gene expression was much reduced in the condition medium treated tumors versus proper controls (p< 0.05). Tissue necrosis was more prevalent in this group while tumors volume was diminished almost by 40%. The LC-ESI-MS/MS analysis unrevealed the stemness reducing and the cell death inducing proteins such as, pigment epithelium-derived factor (PEDF), insulin like growth factor binding protein-5 (IGFBP-5) and -7 (IGFBP-7) in the condition medium. CONCLUSION: This study showed that the substances released from starved human fibroblasts were able to down-regulate the stemness-related genes and induce necrosis in LA7 derived tumors.

In-situ electro-generation and activation of hydrogen peroxide using a CuFeNLDH-CNTs modified graphite cathode for degradation of cefazolin.

The modified multifunctional electrodes for electro-Fenton (EF) process are suggested to be promising cathodes for in situ electro-generation and activation of H2O2 to produce hydroxyl radicals (•OH). However, heterogeneous EF process still faces the challenges of limited catalytic activity and releasing of massive amounts of transition metals to the solution after removal of organic pollutants. The main aim of the present investigation was to prepare a cathode containing carbon nanotubes (CNTs) and CuFe nano-layered double hydroxide (NLDH) for degradation and mineralization of cefazolin antibiotic through electro-Fenton process. Structural and electrochemical analyses demonstrated that CuFeNLDH-CNTs nanocomposite was successfully incorporated on the surface of graphite cathode. Due to the increased formation of •OH in the reactor, the incorporation of CNTs into NLDH matrix with a catalyst loading of 0.1 g substantially improved the degradation efficiency of cefazolin (89.9%) in comparison with CNTs-coated (28.7%) and bare graphite cathode (22.8%) within 100 min. In the presence of 15 mM of ethanol, the degradation efficiency of cefazolin was remarkably decreased to 43.7% by the process, indicating the major role of •OH in the destruction of target molecules. Acidic conditions favored the degradation efficiency of cefazolin by the modified EF process. Mineralization efficiency of the bio-refractory compound was obtained to be 70.1% in terms of chemical oxygen demand (COD) analysis after 300 min. The gas chromatography-mass spectroscopy (GC-MS) analysis was also implemented to identify the intermediate byproducts generated during the degradation of cefazolin in the CuFeNLDH-CNTs/EF reactor.

Targeting LA7 breast cancer stem cells of rat through repressing the genes of stemness-related transcription factors using three different biological fluids.

Down-regulation of stemness genes expression is important in differentiation therapy against cancer stem cells (CSCs). The aim of this study was to evaluate the Oct4 , Sox2, Nanog, and C-myc expression in rat breast cancer stem cells (LA7) which treated with human ovarian follicular fluid (FF), replicative senescent fibroblast culture supernatant (P14), and 16 h serum starved fibroblast supernatant (16 h-SFS). The cells were exposed to these biological fluids for 24 h, 72 h, and 7 days. Stem-loop RT-qPCR assay was used to quantify the expression of above mentioned genes. Results showed that FF had the least cytotoxic effect on the LA7 cells. Except for Nanog gene, exposure of LA7 cell line to 16 h-SFS and P14 decreased significantly expression of the three other genes after 24 h (P < 0.05). Nanog and Sox2 genes expression was also decreased in LA7 cells which have been already treated with FF for 24 h. Moreover, compared to the control solution, the expression of Oct4 increased significantly after 7 days exposure to FF (P < 0.05). Annexin V-PE /7-AAD-, acridine orange/ethidium bromide staining and doubling time assays revealed apoptosis and necrosis induction by these biological fluids in LA7 cells. Moreover, in an in vitro model of metastasis assay, i.e., scratch test, these fluids exhibited anti-LA7 migration activity which culminated in 16 h-SFS treated cells. Generally, this study showed that FF, 16 h-SFS, and P14 have positive effects on down-regulation of Nanog, Oct4, Sox2 and C-myc expression, and consequently can increase the differentiation of breast cancer stem cells. For the first time, this study provided some evidence indicating that some biological fluids have potential to differentiate the CSCs, show anti- survival, growth-, and cell migration activity.

Anticancer properties of chitosan against osteosarcoma, breast cancer and cervical cancer cell lines.

BACKGROUND: Cancer is still the most common cause of morbidity in the world. Chitosan, a commonly used natural polymer, is consisted of different molecular weight with different biological activities.The purpose of this study was to determine cytotoxicity effect of high molecular weight (HMWC) and low molecular weight of chitosan (LMWC) on three cancerous cell lines MCF-7, HeLa and Saos-2 with different histological origin. METHODS: The anticancer property of two types of chitosan on three cancerous cell lines and human fibroblast as normal cell line, was evaluated by cytotoxic activity including their apoptosis induction properties. Chitosan solutions 2% (w/v) were prepared. The cells were treated by different concentration of chitosan and viability was determined by MTT assay after 24, 48 and 72 h .Also the mode of cell death-apoptosis vs necrosis ,was determined by Annexin V staining assay and analyzed by flow cytometry. RESULTS: While both types of chitosan were effective in inhibiting cell proliferation of three cancerous cell lines, fibroblast cells showed somehow more compatibility with chitosan. Despite of a significant decrease in all 3 cell lines viability, up to 90%, but we didn't see a concentration dependent difference between both types of chitosan (HMWC and LMWC) in their cytotoxic effects. Flow cytometry analysis showed necrosis more observable with MCF7 while the apoptosis pattern of death was more in Saos-2 and HeLa. Also, higher viability with both types of chitosan was seen in fibroblast as normal cells. CONCLUSION: While chitosan is compatible with normal diploid fibroblast cells, it shows anticancerous effect against 3 cancerous cell lines. Furthermore, it seems that the molecular weight of chitosan does not affect its anticancerous property.

Template-free microspheres decorated with Cu-Fe-NLDH for catalytic removal of gentamicin in heterogeneous electro-Fenton process.

Nano-layered double hydroxide (NLDH) decorated with Fe and Cu was applied as a novel heterogeneous catalyst for catalytic degradation of gentamicin by the electro-Fenton (EF) process. The EF process was equipped with graphite plate under aeration to electrochemically generate hydrogen peroxide in the solution. The characterization analyses confirmed the suitable structure of as-synthesized Cu-Fe-NLDH to be acted as catalyst for treating the target pollutant. The comparative study showed the highest removal efficiency of 91.3% when the Cu-Fe-NLDH-equipped EF process was applied in comparison with the Fenton (50%) and the electro-oxidation alone (25.6%). The acidic pHs favored the degradation of gentamicin. Increasing the current resulted in the enhanced degradation of gentamicin, while the excessive electrolyte concentration (0.1 mol/L) and catalyst dosage (1.5 g/L) led to the tangible drop in the reactor performance. At a specified reaction time, the injection of O3 gas enhanced the efficiency of the Cu-Fe-NLDH-equipped EF process. The presence of ethanol led to more suppressing effect than benzoquinone, indicating the dominant role of OH radical in the degradation of gentamicin compared with other free radical species such as O2- radical. Only 10% drop in the degradation efficiency of gentamicin was observed within 10 operational runs. The mineralization efficiency of about 77% was achieved after 300 min in terms of chemical oxygen demand (COD) removal. The intermediate byproducts generated during the destructive removal of gentamicin were also identified.

Comparison of Germ Cell Gene Expressions in Spontaneous Monolayer versus Embryoid Body Differentiation of Mouse Embryonic Stem Cells toward Germ Cells.

BACKGROUND: Genetic and morphologic similarities between mouse embryonic stem cells (ESCs) and primordial germ cells (PGCs) make it difficult to distinguish differentiation of these two cell types in vitro. Using specific GC markers expressed in low level or even not expressed in ESCs- can help recognize differentiated cells in vitro. We attempted to differentiate the mouse ESCs into GC-like cells spontaneously in monolayer and EB culture method. MATERIALS AND METHODS: In this experimental study, we attempted to differentiate ESCs, Oct4-GFP OG2, into GC-like cells (GCLCs) spontaneously in two different ways, including: i. Spontaneous differentiation of ESCs in monolayer culture as (SP) and ii. Spontaneous differentiation of ESCs using embryoid body (EB) culture method as (EB+SP). During culture, expression level of four GC specific genes (Fkbp6, Mov10l1, Riken and Tex13) and Mvh, Scp3, Stra8, Oct4 were evaluated. RESULTS: In both groups, Mov10l1 was down-regulated (P=0.3), while Tex13 and Riken were up-regulated (P=0.3 and P=0.04, respectively). Fkbp6 and Stra8 were decreased in EB+SP and they were increased in SP group, while no significant difference was determined between them (P=0.1, P=0.07). Additionally, in SP group, gene expression of Mvh and Scp3 were up-regulated and they had significant differences compared to EB+SP group (P=0.00 and P=0.01, respectively). Oct4 was down-regulated in the both groups. Flow-cytometry analysis showed that mean number of Mvh-positive cells in the SP group was significantly greater compared to ESCs, EB+SP and EB7 groups (P=0.00, P=0.01, and P=0.3, respectively). CONCLUSION: These findings showed that ESCs were differentiated into GCLCs in both group. But spontaneous differentiation of ESCs into GCLCs in SP group (monolayer culture) compared to EB+SP (EB culture methods) has more ability to express GCs markers.

Association study of M235T and A-6G polymorphisms in angiotensinogen gene with risk of developing preeclampsia in Iranian population.

OBJECTIVE: Preeclampsia (PE) is a life-threatening complication of pregnancy that accounts for 12% of all maternal deaths worldwide. The aim of this study is to investigate the relationships between the polymorphisms of angiotensinogen (AGT) gene and preeclampsia. MATERIAL AND METHODS: In this study, 240 unrelated preeclampsia patients and 178 normotensive women were examined. Genomic DNA was extracted then we assessed M235T(C/T) and A-6G polymorphisms of the AGT gene. Genotyping of M235T and A-6G polymorphisms were performed using SSP-PCR and MS-PCR, respectively. RESULTS: A significant protective association was observed between A-6G G allele, A-6G A/G heterozygote genotype (OR = 0.6, p = 0.007 and OR = 0.6, p = 0.04) against PE. Furthermore, it was shown that two copies of A-6G A allele would increase PE risk (OR: 0.62, p = 0.04). Our results did not show a significant association for M235T polymorphism and PE. However, the combinations of A-6G A/A genotype and M235T T/C genotype (OR = 0.4, p = 0.02) and also A-6G A/G genotype and M235T T/C genotype (OR = 0.5, p = 0.04) in controls represented a significant protective association against PE. CONCLUSION: According to the existence of significant correlation between two candidate polymorphisms, A-6G and M235T polymorphisms, with PE disease in our study, they may be considered as valuable factors in susceptibility to PE disease in Iranian women.

Upregulation of miR-371-373 cluster, a human embryonic stem cell specific microRNA cluster, in esophageal squamous cell carcinoma.

AIMS: Esophageal squamous cell carcinoma (ESCC) is the most common subtype of esophageal cancer in Iran. MicroRNAs (miRNAs) are a class of noncoding RNAs that are found to be involved in different processes and can play a role in tumorigenesis and result in cancer. MiR-371, miR-372, and miR-373 are a gene cluster that is located in the region of the human chromosome of 19q13.4. They are specifically expressed in human embryonic stem cells (ESCs) and involved in the maintenance of the stemness features through regulating the expression of certain key genes and signaling pathways. The present study investigated the potential expression of miR-371-373 cluster in tumor and nontumor tissues of ESCC. MATERIALS AND METHODS: The expression level of miR-371-373 cluster was analyzed in paraffin-embedded tissues of tumor and tumor margin in 36 patients with ESCC. Total RNA was isolated and the miR-371-373 clusters were quantified with quantitative real-time-polymerase chain reaction expression analysis. Computed tomography analysis (2-ΔΔCT) and t-test were used to determine the relationship between the characteristics of the tumor and nontumor tissues. Statistically, P value of <0.05 were considered significant. Data analysis was performed using SPSS 16. RESULTS: We provided miR-371, miR-372, and miR-373 upregulation evidence significantly with 14.36, 26.9, and 21.1-fold in esophageal cancer cells compared with their adjacent normal cells (P < 0.05), respectively. In addition, evaluation of these genes expression in various grades didn't show a significant difference. CONCLUSION: Our findings support the hypothesis that these miRNAs might play a role in tumorigenesis in esophageal cancer.

Cytochrome P-450 1B1 Leu432Val Polymorphism Does Not Show Association With Breast Cancer in Northern Iranian Women With a History of Infertility.

The Cytochrome P-4501B1 (CYP1B1) Leu432Val polymorphism has been previously shown to be associated with some types of cancer and affects CYP1B1-mediated metabolism of various infertility drugs. To establish the frequency of CYP1B1 Leu432Val polymorphism among women with a history of infertility drug use, we studied the genotypes of 147 patients with breast cancer with a history of infertility and 150 cancer-free, infertile women (control group) in Northern Iran. A polymerase chain reaction-based restriction fragment length polymorphism assay was used to detect GG (Val/Val), CG (Leu/Val), and CC (Leu/Leu) genotype frequencies, which did not vary significantly between the 2 patient groups (P = .847). We established for the first time that the incidence of CYP1B1 Leu432Val polymorphism is 46.6% among women with infertility history and breast cancer in Northern Iran. Finally, our results do not show any significant association between CYP1B1 Leu432Val polymorphism and breast cancer in infertile women in this region, who have also received infertility treatment.

Detection of Toxoplasma gondii DNA in Malignant Breast Tissues in Breast Cancer Patients.

Breast cancer is the most prevalent malignancy in women throughout the world. Similar to other cancers, a strong relationship between breast cancer and environmental factors such as infectious agents has been reported. Toxoplasma gondii is a protozoan parasite which may play a role in cancer induction. The present study aimed to investigate a possible association between a history of T. gondii infection and breast cancer by detecting T. gondii DNA in malignant and non-malignant breast and lymph nodes tissues from breast cancer patients with latent toxoplasmosis. Formalin-fixed, paraffin-embedded (FFPE) tissue blocks from malignant/non-malignant breast and lymph nodes were obtained from twenty-nine breast cancer patients who were positive for anti-Toxoplasma antibodies (IgG). FFPE tissue blocks were deparaffinized using hot water method, and DNA was extracted. A conventional PCR analysis was performed to amplify partial regions of T. gondii B1 and REP-529 genes. Ninety-three samples from 29 patients were examined. All patients were negative for anti-T. gondii antibodies (IgM). T. gondii DNA was detected in 3 (10.3%) patients by PCR analysis of either B1 or REP-529 genes. These include two malignant breast and one normal lymph node samples. Sequence analysis of these genes showed a good similarity with previously published B1 and REP-529 sequences of T. gondii in NCBI GenBank. This study did not find any association between T. gondii infection and breast cancer. Furthermore, it is the first molecular identification of T. gondii in FFPE tissue samples obtained from breast cancer patients.

Interleukin-1 alpha variation is associated with the risk of developing preeclampsia.

OBJECTIVE: Preeclampsia is a syndrome that affects 5% of all pregnancies, producing substantial maternal and prenatal morbidity and mortality. Several studies have reported that cytokine genes are associated with the persistence of preeclampsia or the severity of the disease. The aim of this study is to investigate the relationships between the polymorphisms of interleukin-1 alpha-889 (IL-1A) gene and preeclampsia. METHOD: Genomic DNA was extracted from the peripheral blood of 305 patients with preeclampsia and 325 normal controls from Sayyad Shirazi Hospital of Golestan University. Then subjected to SSP-PCR amplification. STATA software and the chi square test were used for statistic calculations. RESULTS: The frequencies of IL-1A -889 genotypes C/C, T/T and C/T in preeclampsia cases were 34.8%, 8.2%, 57% and in controls were 20.9%, 7.6% and 71.3% respectively. There was a significant 1.5 fold excess frequency in genotype C/C in cases (CI=1.44-3.07, OR=2.1, P=0.0001). There was a significant difference in the frequencies of alleles or genotypes in IL-1A promoter regions between patients with preeclampsia and the control group. Turkomans showed the highest frequency of the C allele and Sistanies had the lowest frequency of the C allele in preeclampsia compared to control groups (CI=1.5-3.9, OR=2.48, P=0.0001). CONCLUSION: Our findings suggest that the IL-1A-899C/C genotype and C allele are associated with susceptibility to preeclampsia.