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The flowering process in plants proceeds through the induction of an inflorescence meristem triggered by several pathways. Many of the genes associated with these pathways encode transcription factors of the MADS domain family. The MADS-domain transcription factor APETALA1 (AP1) is a key regulator of flower development. The first step to understand the molecular mechanisms under the function of each gene in a plant is identification, sequencing and phylogeny analysis of that gene. For this purpose, total RNA was isolated from flower bud of garden cress (Lepidium sativum L.) and was used for cDNA synthesis. The specific primers were designed based on nucleotide sequence alignment of AP1 homologus genes from plants of the same family Brassicaceae and were used in RT-PCR. After observing its electrophoretic pattern and ensuring the quality of PCR product, the amplicon was sent for sequencing. After receiving the results of sequencing, the sequence examined with BLAST, MUSCLE, Gene Runner and MEGA6 softwares. The results indicated amplification of 787 nucleotides fragment that named LsAP1 and was recorded by accession number KP070728 in NCBI database. The studies show high similarity and overlapping of gene bank sequences with LsAP1 illative protein. According to these results, LsAP1 may play a similar role as AP1 in flower induction and could act as a flower meristem identity gene in Lepidium sativum L.

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Plant peroxidase (EC: 1.11.1.7) a heme-containing protein which is widely used in plants, microorganisms and animals. This two - substrate enzyme, catalyze the hydrogen peroxide into water with   oxidation of many organic and inorganic substrates that all of them can be used to measure enzyme activity. Although it’s specific substrate is hydrogen peroxide. Calcium and at least four disulfide bonds in the protein structure lead the formation and strength of three-dimensional structure of the molecule. Plant peroxidase has several roles including, involvement in lignin biosynthesis, auxin metabolism, cell growth, cell wall cross linking and respond to environmental stress. So peroxidase, considered as a good point to pursue the cell deal with stress factors such as oxidative stress. These days according to produce the pure samples of this molecule, peroxidase also used in ligand-protein studies in pharmaceutical research. So in this brief overview, in addition to introducing plant peroxidase we have had a brief look to measure the enzyme activity, the number of isoenzymes in a cell and the ensuing conformational changes of peroxidase.

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In this study, the cytotoxic effects of aqueous and ethanolic extracts of Sedum album L. on human stomach cancer cell line (AGS) and breast cancer cell line (MCF-7) were evaluated by MTT, BrdU and TUNEL assays. The results demonstrated that both extracts had antiproliferative and apoptotic effects in a dose-dependent manner. The MTT assay data revealed that the AGS cell underwent more cytotoxicity in comparison with the MCF-7 cell. It also revealed that ethanolic extract was more potent than aqueous extract. The BrdU assay results showed that the proliferation of AGS and MCF-7 cells was reduced to 50% and 43%, respectively, at the highest concentration of the aqueous extract. In addition, the ethanolic extract reduced the proliferation of AGS and MCF-7 cells to 75% and 60%, respectively. The AGS and MCF-7 cells underwent 52% and 12% apoptotic death upon treatment by the ethanolic extract as TUNEL assay showed. The aqueous extract induced 28% and 25% apoptosis in the AGS and MCF-7 cells, respectively. Both inhibition of proliferation and induction of apoptosis are desirable strategies for cancer treatment among researchers. Identification of S. album compounds and analyzing their effects in animal model of cancer can help us with understanding its anti-cancer properties.
 

 

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In recent years using of silver nanoparticles due to its unique properties was increased. Medical plant, the Achillea biebersteinii is rich in anti-cancers compounds. The aim of this study was to investigate the cytotoxicity effects of AgNPs synthesized using Achillea biebersteinii extract on human ovarian cancer A2780 cells. Cytotoxic effects of AgNPs with MTT test was performed at 48 hours in concentration of 2, 4, 6, 8, 16 and 32 µg/ml. To study the cell death induced by AgNPs DAPI, acridine orange (AO)/ Propidium iodide (PI) staining and Annexin v/ Propidium iodide assay and activation of caspase 3 and 9 were assessed. Results: The results showed that the synthesized AgNPs decreased cell viability dose dependently, calculated inhibitory concentration of 50 % (IC50) was 4 µg/ml.  The results from AO / PI, DAPI staining, Annexin V / PI showed that in treated cells the percentage of apoptotic cells compared with control increased. In addition, the treated cells showed increased activation of caspase 3/9. Hence silver nanoparticles induced cell death through the caspase dependent pathway. The results suggest that AgNPs synthesized using Achillea biebersteinii   extract exert their cytotoxic effect by inducing apoptosis.

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Catechin, epicatechin gallate (ECG) and quercetin, as bioactive flavonoids, have been shown to possess anticarcinogenic effects. Ceramide plays an important role in killing tumor cells. Accordingly, the aim of this study was to clarify the involvement of these compounds in ceramide metabolism in A549 cancerous cell line. Spectrophotometer, cell culture and HPLC methods were used. Cell viability index showed different potential of cytotoxicity effect for each of the studied agents, among which ECG was more potent. This index decreased significantly over 100 to 250 µM concentrations of treatment with respect to control. Cell treatments also caused considerable increase in ceramide level within cells in a dose-dependent manner. Sphingomyelinase activity increased significantly in treatment with quercetin and catechin. There was significant inhibition in acid ceramidase activity of cell extract in response to each of the three compounds, particularly over 100 µM in comparison with control. Data also showed no significant variation in glycosyl ceramide synthase activity in treated cells with quercetin, whereas the activity decreased significantly by Catechin and/or ECG. It is our conviction that different effects on ceramide metabolism enzymes may be related to various chemical groups on the common structure of the studied compounds. Due to structure-function relationship, these compounds had different effects on ceramide generation. Elevation in ceramide content in A549 cancer cell line induced apoptosis, which led to anti-cancerous effects, as observed in this study.

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Yarrowia lipolytica, as a good cell factory to speed up the production of plant pharmaceutical components, has been considered to be one of the most important and attractive micro-organisms in recent years, due to its high secretion capacity, limited glycosylation, large range of genetic markers and molecular tools. Naringenin, as a central core of flavonoids production, plays important roles both in plants and in the treatment of different types of human diseases. For this purpose, specific naringenin biosynthesis genes from different origins were selected and introduced after comparative expression profiling in Y. lipolytica. This research indicated that chs plays the main role in the production of naringenin, so the increase copy number of this gene in each construct was investigated. The HPLC results confirmed that the construct with 5 copy numbers of chs resulted in 7.14 fold increase of naringenin extracellular titer to 90.16 mg/L in shake flask cultures. The results reported in this study demonstrated that sufficient knowledge of genes involved in the specific biosynthesis pathway, synthetic gene pathway and using Y. lipolytica as a capable and cheap host could help bioengineers to produce significant amounts of pharmaceutical components.
 
 

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Superparamagnetic iron oxide nanoparticles (SPIONs) have made extensive advances in nanotechnology. The unique properties of these particles have expanded their application in various fields, including medicine. One of these applications is non-invasive analysis for cell tracking. However, the possibility of toxicity in cells is reported by these nanoparticles. Due to the fact that cellular damage caused by iron oxide nanoparticles is concentration-dependent, the determination of the appropriate  concentration of iron oxide nanoparticles is very important to prevent cell damage or cell death due to apoptosis. The aim of this study was to find a concentration of SPIONs which does not result in apoptosis. Therefore, the effects of different concentrations of iron oxide nanoparticles on cell survival were investigated, and the their effects on increased gene expression involved in apoptosis (p53) in human amniotic membrane derived mesenchymal stem cells (hAMSCs) were evaluated. First, stem cells were extracted from human amniotic membrane tissue and cultured. To demonstrate the multipotent characteristic of hAMSCs, these cells were differentiated into adipose, bone, and chondrocyte cell lines. Then, the viability of the cells treated with different concentrations of iron oxide nanoparticles (200, 150, 100, 50, 0 μg / ml) over a period of 24 and 48 hours was evaluated by MTT method. The effect of the concentrations of 0, 100,150 and 200 μg / ml of nanoparticles after 24 hours in hAMSCs was investigated for the expression of p53 gene by Real-Time PCR. hAMSCs were spindle-shaped in a two-dimensional culture. Flow cytometry examination of surface markers revealed that these cells were able to express CD 29, CD90 and CD105 but they were unable to express CD34 and CD45. The results of the multi-potency assay of hAMSCs showed that these cells were capable of being differentiated into adipocyte, bone and chondrocyte cell lines. Iron oxide nanoparticles had no significant effect on cell survival at the concentrations of 50 and 100 μg / ml in 24 hours. However, cell viability decreased significantly after the concentration of 150 μg / ml (42 ± 1.4%, p<0. 001. The results of Real-Time PCR  analysis showed that the expression of p53 gene significantly increased at concentrations of 150 (2.4±0.1, P < 0. 001) and 200 μg / ml (4.1 ± 0.11, P < 0. 001). According to the results, the nanoparticles used in this study were appropriate at concentrations ≥ 100 μg / ml for cell tracking.

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Tetanus is caused by the toxin secreted by Clostridium tetani. Due to the rapid infection with this bacterium, it is so important to investigate the tetanus immunity of people. Therefore, electrochemical biosensors, as one of the most effective tools in this regard, have demanded characteristics such as being fast, simple, cost-effective and portable. However, their detection sensitivity is not sufficient. Hereon, silver enhancement of gold-nanoparticles was proposed for the improvement of detection. Hence, the current study applied gold-nanoparticles as label, following with silver enhancement, to investigate the yes/no electrochemical detection of anti-tetanus toxoid antibodies in the indirect immunoassay utilizing glassy-carbon electrodes modified by carbon nanotubes. The analytical procedure consists of the reactions of the tetanus toxoid with the antibody at electrode, so that followed by the interaction of gold-labeled secondary antibody and then silver enhancement process. In this study, the cyclic-voltammeter variation and difference of gold to silver signal based on silver ions fluctuations were also investigated. The results indicated that ∆E_p increased from 0.24 V before silver enhancement reaction to 0.57 V after the silver enhancement. The results also demonstrated that after silver enhancement, current significantly increased and current plot at E^c_p transferred to positive potentials and at E^a_p moved to negative potentials. In conclusion, this method increases the detection sensitivity and can simply use to other bio-molecules detection.
 
 

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Exosomes are secreted by different types of cells and known as biological packages. Exosomes have significant role in intercellular communications and involved in the development and progression of various diseases such as cancer. Inhibin B and anti-mullerian hormone (AMH) are markers of granulosa cell tumors (GCT) and due to the role of exosomes in the progression of cancer, in this experimental study, the effect of exosomes derived from human ovarian cancer cells on the secretion of Inhibin B and antimullerian hormone (AMH) by granulosa cells was investigated. First, A2780 human ovarian cancer cells were cultured, then the supernatant was collected to extract the exosomes by ultracentrifugation and subsequently, the extracted exosomes were checked out using dynamic light scattering (DLS) and Scanning electron microscopy (SEM). In addition, granulosa cells were isolated and cultured from the ovaries immature female Balb / C mice and treated with 25 μg/ml of exosomes derived from the ovarian cancer cell-line. Inhibin B and AMH hormones levels were then measured. The results showed the significant (P<0/05) increase of the level of inhibin B and AMH hormones in the treated cells in comparison with the control group. According to the results, exosomes increased the secretion of Inhibin B and AMH hormones and seems to be effective in the proliferation of granulosa cell tumors.
 
 

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Nanotechnology-based targeting drug delivery systems have a considerable potential in medicine. Therefore, the present study aimed to designe, synthesise and characterize a nanodrug with beta lactoglobulin coating including oxali-palladium with and without folate and to compare their anti-cancer effects. The physicochemical properties of nanocapsules were studied by Dynamic light scattering (DLS), atomic force microscopy (AFM) and scanning electron microscopy (SEM). Finally, the anticancer activities of nanodrugs were investigated against human colorectal cancer cell line of HCT116 by MTT and flowcytometry methods. The results of Dynamic light scattering (DLS), scanning electron microscopy (SEM) and atomic force microscopy (AFM) showed that the average size of nanocapsules with folate were less than 40 nm. Cytotoxicity results proved the dose- and time-dependent antiproliferation and anicancer activities of nanocapsules (with folate) against HCT116. Finally, it could be concluded that folate increase anticancer activity of nanodrugs and it might be considered as a new candidate in the design and synthesis of new drugs in cancer treatment.


 

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Lung carcinoma is the second most common type of cancer. Inefficiency of the current treatments and the undesirable side effects of chemotherapy drugs made the know-how of the treatment important. The purpose of this study is to investigate the synergic effect of curcumin and Cisplatin in comparison with the sole application of each treatment on Calu-6 cell line, an epithelial cell line of human lung carcinoma, and the expression of Cdc42 gene. The viability of Calu-6 was examined after 24- or 48-hour treatment with doses of 0.5 to 8 µg/ml of curcumin, 0.1 to 50 µg/ml of cisplatin and combined doses of curcumin and Cisplatin by MTT assay. To measure apoptosis and the expression of Cdc42 gene, flow cytometry and Real-Time PCR were utilized. Decrease of cell viability and induction of cell death were observed in the cells treated with 0.67 µg/ml of curcumin and 1.7 µg/ml of cisplatin (the lowest effective dose) and the combined treatment with the same doses of each drug after 24-hour treatments. The maximum rates of early and late apoptosis were related to treatment with curcumin and the combined treatment. The gene expression analysis results indicated that both Curcumin and Cisplatin decrease the expression of Cdc42 gene, moreover, their co-administration showed synergic effects. Therefore, Curcumin could be an appropriate option for complementary administration with other chemotherapy agents in order to reduce their efficient dose, and to reduce their side effects.
 
 

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This study aimed to investigate the polymorphism of matrix metalloproteinase -3 (MMP-3) gene and its expression in the serum of infertile female patients received in vitro fertilization and embryo transfer (IVF-ET). To do so, 100 women with unsuccessful IVF-ET (IVF^–) and 100 women with successful IVF-ET procedure and clinical pregnancy (IVF⁺) were included. Genetic polymorphism and serum concentration of MMP3 were investigated by ARMS-PCR and ELISA, respectively. The results showed no significant association between MMP-3 gene polymorphism and IVF-ET outcome among the two groups studied. However, a significant decrease in the concentration of MMP-3 serum in the IVF^– group was observed in comparison with the IVF⁺ group (P=0.000002). Moreover, we showed that the serum MMP-3 levels in CC, AC and AA genotypes in the IVF^– group were 33, 65.33 and 86 ng/ml, respectively. In conclusion, while there is no significant difference between MMP-3 promoter polymorphism and IVF-ET outcome between the IVF⁺ and IVF^- groups, a significant decrease in MMP-3 serum levels in IVF^- group was seen as compared with the IVF⁺ group. It could be also suggested that the CC genotype is associated with a decreased level of MMP-3 serum concentration and may be associated with IVF-ET failure.
 

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Peroxidase catalyzes different oxidation of substrates using hydrogen peroxide, a reactive oxygen specie (ROS). ROS, at low concentrations, act as messenger to regulate intracellular signaling, whereas, at high concentrations, they can overcome the immune system by creating oxidative stress. Some common beverages such as coffee, tea and soft drinks contain high levels of xanthine alkaloids including theophylline and theobromine. In this study, the effect of theophylline and theobromine on peroxidase activity was kinetically studied by measuring the absorption of 4-aminoantipyrine, oxidized in the presence and absence of theophylline and theobromine at 510 nm for 3 minutes. The results showed that theobromine and theophylline acted as inhibitors with IC₅₀ of 0.50 and 0.55 mM, respectively. K_m and V_max values showed that both compounds are non-competitive inhibitors. The values of K_i were calculated as 0.03 and 0.045 mM for theobromine and theophylline, respectively. Lower values of K_i and IC₅₀ for theobromine compared to theophylline indicates that theobromine has a higher inhibition strength and binding tendency to the enzyme-substrate complex. Hence, it is concluded that theobromine has a stronger inhibitory effect on POD activity.
 
 

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Human serum albumin is one of the most important blood proteins that has the ability to bind a wide range of compounds and different drugs. Hence, knowing how drugs bind to albumin is crucial to understand their pharmacokinetics and pharmacodynamic properties. The binding of drugs to protein affects the drug's excretion, distribution and interaction in the target tissues. Nicotinamide (NA) is a safe and inexpensive medical supplement that used to prevent and treat vitamin B3 deficiency. In this research, the molecular mechanism of the interaction between nicotinamide and human serum albumin was studied by the utilization of spectroscopic and molecular docking methods. The effects of temperature, acidic/basic pHs, metal ions, urea, and glucose on the interaction between nicotinamide and human serum albumin were also investigated. The spectroscopic studies indicated that the interaction between nicotinamide and human serum albumin is mainly controled by hydrophobic forces and the interaction is spontaneous. The number of binding site and binding constant is 1 and 4.6×10⁴ (L/mol), respectively, which were increased in the presence of glucose. The presence of metallic ions and basic pH decreased the binding constant of nicotinamide to albumin. The obtained results indicated that nicotinamide tend to binds to the similar sites wherever the molecules with acidic moieties bind. The results could be helpful to interpret the mechanisms of actions of nicotinamide in the various physiological phenomena in the human body.

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This study aimed to identify PGP and MRPA genes in clinical isolates of Leishmania. The genes of pgpa (MRPA) and mdr1 (PGP) are involved in the drug resistance, their products act as dependent transporters of ATP (ABC Transporter) in the reflux of drugs from the cytosol to the outer space of the cell. Hence, 40 volunteers with leishmaniasis were randomly selected. Firstly, Amastigotes were examined under a light microscope, then inoculated into NNN-specific biphasic culture medium. Deoxy ribonucleic acids were extracted by phenol-chloroform method and were determined by ITS-specific primers. Then the frequency of two pumps involved in "drug resistance" was investigated by PCR. In this study, the mdr1 gene, which had previously been shown to be present in the in vitro resistant strains, was shown to have a higher frequency of pgpas, which could be due to the presence of MDR. It transports the drug from the inner layers of the lipid bilayer membrane to the outer layers, reducing the concentration of the drug inside the cell and causing drug resistance, while the MRPA pump is in the membrane of the cell organelles.

 

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Cisplatin, as a chemotherapy drug, causes serious side effects in the advanced stages of the cancer. Recently, Artemisia has been considered for its bioactive compounds, anti-proliferative and anti-inflammatory effects. The aim of this study was to evaluate the anti-cancer and anti-metastatic effects of the methanolic extract of aerial organs of Artemisia and cisplatin, either alone or in combination, in human ovarian cancer cell line A2780. The viability of A2780 cells after treatment with Artemisia extract, cisplatin and their combination was evaluated by MTT assay and the alterations in the morphology of the cell nuclei were examined by DAPI staining. The induction of apoptosis was assessed by Annexin V test, cell migration and changes in expression levels of apoptotic genes (Bax and P53) and metastasis (MMP2 and MMP9) using real-time PCR. MTT test data showed that Artemisia extract, cisplatin and their combination decreased the viability of ovarian cancer cells. DAPI and Annexin V indicated the DNA fragmentation and increased percentage of cellular apoptosis in comparison with the control group. The migration and real-time PCR data showed a decline in thr cell invasion and expression of genes involved in metastasis (MMP2 and MMP9) in cancer cells while the expression of apoptotic genes (Bax and P53) was increased in the treated groups. The results of this study showed that while both Artemisia extract and cisplatin posses anti-proliferative effect, apoptotic and suitable anti-metastatic effects on their own in A2780 cell line, their combination have synergic effects and posses those desired properties in lower concentration of cisplatin, which can reduce the side effects of cisplatin in cancer treatment.

 

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Pancreatic cancer is one of the most deadly and aggressive cancers; Fluorouracil induces apoptosis and cell cycle arrest in cancer cells. In the present study; the effect of Fluorouracil on different stages of the cell cycle and the expression of genes involved in the internal pathway of apoptosis in the AsPC-1 cell line (human pancreatic cancer) were investigated. In order to do so, MTT assay was used to evaluate the cytotoxic effect of Fluorouracil on AsPC-1 cell proliferation; The type of induced cell death and cell cycle changes were investigated by flow cytometry; changes in the expression level of genes (BAX, Bcl-2, APAF-1, Caspase-3, Caspase-9, p53, p21) were examined by Real-time PCR. Quantitative data were analyzed at the significant level of (p<0.05). The MTT assay results showed that Fluorouracil decreased AsPC-1 cell proliferation in a concentration-dependent manner. The results of flow cytometry analysis showed that increased percentage of apoptotic cells in the treated cells; Fluorouracil induces S phase cell cycle arrest in AsPC-1 cells and reduced distribution in the G1 phase. The Real-time PCR results in treated cells showed an increase in the expression of genes in the mitochondrial apoptotic pathway as well as genes effective in regulating the cell cycle. Fluorouracil reduces cell proliferation and induces apoptosis by increasing the expression of genes involved in the Intrinsic apoptotic pathway in AsPC-1 cells; Fluorouracil also caused cell cycle arrest in these cells by regulating the (p53, p21) genes.
 

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It has been found that the second isoform of COX enzyme known as COX-2 plays an important role in inflammation and rheumatoid arthritis and osteoarthritis. Thus, designing COX-2 inhibitors to treat inflammation is among the most important goals of researchers. In this study, the inhibitory effect of 3 new imidazole derivatives on COX-2 was evaluated by in silico approach. Molecular docking was done using Autodock Vina and the best binding mode of inhibitors was used as input of molecular dynamics (MD) simulation. MD was performed using Gromacs software for 120 ns. Then, structural and thermodynamic analyzes (ΔG_binding) and prediction of physicochemical properties were performed. RMSD data showed the compounds reached a good equilibrium and had favorable stability during simulation. Also, the RMSF showed that due to binding of inhibitors, the fluctuations of complexes decreased and the active site residues had the lowest amount. Rg, SASA and DSSP analysis showed that the protein structure did not change significantly. It was also found that Ser530 and Tyr355 residues play a more effective role in hydrogen bond formation. Physicochemical parameters determined the good drug-likeness properties for all compounds. Structural and thermodynamic analyzes (MM-PBSA) and IC₅₀ data indicate the favorable inhibitory effect of compound 5b.