parvaneh maghami

Gonadotropin hormone-releasing hormone (GnRH) is a peptide involved in mammals’ fertility and may also serve as a candidate target for cancer immunotherapy. Immunonsterilization is known as a proper alternative to surgical castration and has been advocated by European countries in recent years. Immunization with GnRH can effectively inhibit the secretion of gonadotropins and cause infertility in both genders of mammals. In this study, a recombinant trimer of GnRH is designed and expressed in a prokaryotic system.  A construct containing GnRH trimer was designed and prepared using gene synthesis. A cloning site was embedded and connected to the GnRH using a linker to further clone any protein of interest. The construct was subcloned into a pET-32a+ plasmid vector. The recombinant vector was transferred to BL21, an Escherichia coli strain, and the expression was induced using isopropyl β- d-1-thiogalactopyranoside (IPTG). The cell lysate was prepared using lysis buffer and nickel affinity chromatography purification. The GnRH expression was evaluated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis after protein purification. The cloning was verified using a polymerase chain reaction (PCR) followed by sequencing the recombinant vector. The result of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis verified the recombinant protein’s expression and the purification process’s function. The GnRH was properly cloned and expressed in BL21. The results also verified the reliability of the purification process. The construct design allows the researchers to express another peptide sequence attached to the GnRH using the embedded linker to improve the stability and antigenicity. A stable recombinant GnRH would be applied in immunocastration and cancer immunotherapies.

L-Asparaginase converts L-asparagine to L-aspartic acid and causes cancer cells to starve. The main idea of the current study was to improve the biochemical properties of this enzyme using immobilization onto modified magnetic nano-particles (NPs). To this end, Fe3O4 NPs were synthesized, coated with an Au shell, and conjugated with cysteine. The formation of NPs and core-shell structures and their morphology were confirmed using Fourier Transform Infrared spectroscopy (FTIR), Energy Dispersive X-Ray (EDX), VU-Vis, Scanning Electron Microscopy (SEM), and Transmission Electron Microscopy (TEM). Also, Circular Dichroism (CD) and fluorescence spectroscopy were employed for the analysis of the secondary and tertiary structures of the immobilized L-ASNase. The alterations in the kinetic parameters of the immobilized enzyme were analyzed using a Lineweaver-Burk plot. The results of instrumental chemistry analysis confirmed the formation of NPs and core-shell structure, and cysteine binding with the core-shell. Based on CD and fluorescence results, no significant changes were observed in the secondary and tertiary structures of the immobilized enzyme compared to the free one. Kinetic parameters of the immobilized enzyme improved compared to the free enzyme so that Km decreased from 4.43±0.05 to 3.75±0.12 mM and Vmax increased from 187.23±11 to 224.78±16 μM min-1mg-1. Also, the stability of the immobilized enzyme improved with acidic and alkaline pH values compared to the free one at temperatures higher than 50 0C. In addition, the reusability of the immobilized enzyme was superior to the free enzyme, with the immobilized enzyme maintaining 72% of its activity after 15 cycles of catalytic reaction. The immobilized enzyme showed an 86% residual activity after 120 min incubation with trypsin, which was higher than the free enzyme (37%). According to the results of this study, immobilization of L-ASNase onto magnetic NPs can be an efficient strategy to enhance the biochemical properties of this enzyme.

Adsorption of protein on inorganic surfaces leads to structural and functional changes that depend on the nature of the adsorbed protein and the physicochemical properties of inorganic surfaces. Magnetic nanoparticles (MNPs) with biocompatible coatings are the only FDA-approved nanostructured materials with various applications in medical sciences and unique properties have become a high potential material. Study of the bioimmunity of MNPs requires detailed knowledge of the interaction of MNPs with biological molecules, especially proteins. The present study, Comparison of the effects of functionalized MNPs by mesoporous silica and L-lysine (NH2-Si@Fe3O4) on structural changes of egg white lysozyme protein (HEWL).

Structural properties and physicochemical characteristics of functionalized MNPs by FTIR, XRD, TEM, FESEM, VSM and DLS analyzes and Protein-nanoparticle interaction was performed using intrinsic fluorescence techniques, ANS test, thioflavin T (ThT) and ATR-FTIR and protein thermal aggregation. Structure change of HEWL on interaction with MNPs was studied.

Investigation of the interaction of HEWL and MNPs with the help of intrinsic fluorescence showed that the binding of proteins to MNPs is influenced by the type of ligand attached to MNPs. Screening thermal aggregation of HEWL in the presence of MNPs revealed the effective role of NH2-Si@Fe3O4 in inhibiting thermal aggregation of HEWL.

Comparison of nanoparticle performance with functional group and without functional group showed that functionalized nanoparticles had better performance; therefore, it is important to consider the environmental, health and safety aspects in the early stages of nanoparticle use. This report provides an integrated picture of protein-MNPs interaction and the change in protein structure after binding to MNPs.

Primordial germ cell (PGCs) lines are a source of a highly specialized type of cells, characteristically oocytes,during female germline development in vivo. The oocyte growth begins in the transition from the primary follicle. It isassociated with dynamic changes in gene expression, but the gene-regulating signals and transcription factors that control oocyte growth remain unknown. We aim to investigate the differentiation potential of mouse bone marrow mesenchymal stem cells (mMSCs) into female germ-like cells by testing several signals and transcription factors. In this experimental study, mMSCs were extracted from mice femur bone using the flushingtechnique. The cluster-differentiation (CD) of superficial mesenchymal markers was determined with flow cytometric analysis. We applied a set of transcription factors including retinoic acid (RA), titanium nanotubes (TNTs), and fibrin such as TNT-coated fibrin (F+TNT) formation and (RA+F+TNT) induction, and investigated the changes in gene, MVH/ DDX4, expression and functional screening using an in vitro mouse oocyte development condition. Germ cell markers expression, (MVH / DDX4), was analyzed with Immunocytochemistry staining, quantitative transcription-polymerase chain reaction (RT-qPCR) analysis, and Western blots. The expression of CD was confirmed by flow cytometry. The phase determination of the TNTs and F+TNT were confirmed using x-ray diffraction (XRD) and scanning electron microscope (SEM), respectively. Remarkably, applying these transcription factors quickly induced pluripotent stem cells into oocyte-like cells that were sufficient to generate female germlike cells, growth, and maturation from mMSCs differentiation. These transcription factors formed oocyte-like cells specification of stem cells, epigenetic reprogramming, or meiosis and indicate that oocyte meiosis initiation and oocyte growth are not separable from the previous epigenetic reprogramming in stem cells in vitro. Results suggested several transcription factors may apply for arranging oocyte-like cell growth and supplies an alternative source of in vitro maturation (IVM).

In this study the effect of C. colocynthis extract and phycocyanin on the growth of colon cancer cells, and the expression of Caspase 3 and Bax were investigated.Materials &

colon cancer cells (HT29) were treated with different concentrations of C. colocynthis and phycocyanin extracts for 24, 48 & 72 hours. The percentage of cell survival was then measured by MTT assay. Expression of Caspase 3 and Bax genes was investigated by real-time PCR. At the end results were analyzed by SPSS software.

Significant differences were observed in the average percentage of dead cells with 2 to 30 g/μL concentration of extract, phycocyanin, extract + phycocyanin after 24, 48 and 72 hours (p<0.05). The combination of extract and phycocyanin as well as phycocyanin alone showed stronger inhibitory effects on growth of cancer cells compared to extract (p <0.05). The expression of Bax gene was significantly increased by treatment of combination of extract and phycocyanin (2.55-fold) and also C. colocynthis extract alone (1.67-fold) (p <0.01). In addition, the combination of extract and phycocyanin and Abujahl watermelon extract significantly increased the expression of caspase 3 gene (2.15 and 1.75), respectively (p <0.01).

The anticancer effect of Citrullus Colocynthis extract, as well as phycocyanin can be applied by increase the expression of Bax and caspase 3 genes and as a result, apoptosis induction in cancer cells.

Many of traditional infertility treatment methods due to lack of gamete are ineffective.  Differentiation of stem cells to gamete germ cells may be a good choice for producing gametes for treatment of infertility in clinique. Bone marrow mesenchymal stem cells (BMMSc) were obtained from femoral bone of adult NMRI mice by flushing method. Nano tube particles of titanium (TNT), retinoic acid and UV beam 254 ± 20 nm frequencies were used for differentiation of MSc to gamete germ cells. Ultrastructure of TNT was studied with FEM SE electron microscope. Fibrin coating of Nano tubes was evaluated by SEM. Nature and character of stem cells were measured using cluster– differentiation (CD) by flow cytometry confirmed by CD90, and CD31 markers.  Finally, MVH marker was used for assessment of differentiation of MSc to gamete germ cells with immunohistochemistry assay. Statistical data was analyzed by ANOVA and Mann-Whitney tests. Electron microscopic pictures by FE SEM showed homogenous ultrastructure of titanium nanotubes in 10-15 nm diameters. Coating of TNT by fibrin was confirmed by SEM. MTT assay showed most survival rate of MSc cells in 50 µg/ml concentration of TNT. By CD90 and CD31 markers, nature of the stem cells was controlled by flow cytometry. Finally MVH marker expression showed differentiation of MSc to gamete germ cells in treated as compared to control groups and was confirmed by immunohistochemistry. Application of TNT fibrin coated and UV beam can increase differentiation rate of BMMSc to gamete germ cells significantly as a clinical protocol.

The aim of this study was to investigate the effect of Cartap hydrochloride on the structure and function of hemoglobin to determine the molecular mechanism of toxicity of toxin. In this research, the effect of Cartap on the structure and aggregation of human hemoglobin was investigated by UV-visible spectroscopy, thermal spectroscopy, florescent spectroscopy and Attenuated total reflection Fourier transform infrared (ATR‐FTIR) spectroscopy in both titration and incubation conditions and in different concentrations of toxic. Based on UV-visible spectroscopy, results revealed that absorption spectra were changed by adding different concentrations of toxin. Analysis of the levels of oxy-deoxy and met hemoglobin in presence of various concentration of Cartap showed a decrease in active hemoglobin species. Moreover, the aggregation and hemolysis of red blood cells increased in presence of Cartap. In florescent spectroscopy minor degradation of Heme happened and in the second structure of hemoglobin has not altered in presence of toxin.

Antioxidants are of great importance because they can protect the body from oxidation agents. Reactive oxygen species (ROS) are constantly producing as a result of metabolic processes. However, environmental factors such as methyl tert-butyl ether (MTBE) can enhance oxidation stress. MTBE is a widely used fuel oxygenation liquid that has been shown to cause potential cancer. The use of antioxidants may help to reduce the oxidation condition caused by MTBE. Selenium is an essential element with antioxidant property. In this study, the interaction between Cyt C and MTBE has been investigated in the absence and presence of Se. Molecular level examinations are extremely important to investigate the structural change of proteins by MTBE. In this research, molecular behaviour of Cyt c as an important model of protein, in the presence and absence of MTBE and Se is detected by biophysical methods such as UV-vis, fluorescence and FTIR spectroscopy, chemiluminescence and molecular docking. The results showed that MTBE is able to alter Cyt c tertiary structure by ROS production with no change in secondary structure and Se could not protect the protein from structural perturbation. Molecular docking was also confirmed by the prominent interaction of Cyt c with MTBE and Se at a different site. Then Se can cause the structural change of Cyt c.

According to the formation and evolution of life along with static magnetic fields,the permanent exposure has given adaptive ability to beings. Therapeutic magnetism is one of the branches of complementary medicine which uses  the low intensity and non-harmful magnetic fields to the body. By studying in infertile couples (20% male factor), the only cause of infertility and in 50% of cases it is considered as an intermediate factor. One of the influential factors in infertility in men is sperm. In the present study, normal specimens in the magnetic field under the intensities of 1,6 and 12 millitesla and at 1,3 and 5 h intervals. Sperm movement rate was evaluated by CASA, as well as sperm viability, by eosin staining of necrosin and morphology by staining Papanicula. The results of this step on normal sperm showed a significant reduction in the sperm movement ,which  that was not affected by the field. Morphological studies also show that sperm motility is not affected by magnetic field.. the survival rate of sperm  was affected by the magnetic field was significantly reduced, and the sperm morphology remained unchanged

Colorectal Cancer is one of the most common malignancies and is one of the mojor causes of mortality. Todayes, researchers pay attention to use of herbal extracts, including phycocyanin and Citrus colocynthis extract for treatment of colon cancer. In this study the synergic effect of these compoundson colon cancer cells growth and expression of caspase-8 and Bcl2 genes was assessed. After treatment of human colon cancer cells (HT-29) in differnet times (24, 48 & 72h) and with different concentrations of aqueous and phycocyanin watermelon extract and also combination of aqueous extract and phycocyanin, percentage of live cells was calculated using MTT method. Then expression of caspase-8 and Bcl2 genes were measured by real-time PCR. The highest inhibition of growth of cancer cells was observed after treatment with high concentration of both compounds. In addition, significant differences were observed between the different treatment times with low concentration (1 and 2 g/ml). The aqueous extractand phycocyanin reduced the expression of Bcl2 gene by about 10.27 and 5.22, respectively. However, the combination of extract and phycocyanin was associated with a decrease of 7.31% in expression of this gene. The extract increased the expression of Caspase-8 gene by 2.3 fold. Citrus colocynthis extract induces apoptosis by strongly increasing the expression of caspase-8 gene. Additionally, the phycocyanin and the combination of extract and phycocyanin strongly decreased the expression of Bcl2 gene. Phycocyanin and extract and Phycocyanin combination had stronger anti-cancer effects than extract alone.

Benzene is one of the major pollutants in the air and a chemical compound of the aromatic hydrocarbon group, which is widely used in the industry. Exposure to benzene can lead to biologic damages, especially cancer, as it has been reported in many studies of benzene mortality. In this study, we examined the interaction between benzene and lysozyme enzyme which plays a major role in non-specific immunity in the body and also is a natural antibiotic. Structural changes resulting from the interaction of benzene and egg white lysozyme by spectroscopic methods showed that the protein 3D structure changed in the presence of different concentrations of benzene. Changes in the protein function will induce multiple consequences in the biological phenomena at the cellular levels. Lysozyme extrinsic fluorescence was also indicated a significant increase in the presence of benzene. The results of the chemiluminescence method suggested that, with increasing benzene concentration, the production of free radicals was increased. Therefore, the destructive effect of benzene on the lysozyme structure is due to the production of reactive oxygen radicals. Because of the relationship between structure and function in proteins, benzene plays a major role in its function by changing the protein structure. By measuring the activity of enzyme in the presence of different concentrations of benzene, the lysozyme function was reduced.

Extensive use of methyl tert-butyl ether (MTBE) has raised significant threats to the environment through pollution of environmental resources including ground waters. This compound could accumulate in the blood stream through inhalation of contaminated air since MTBE has a high affinity for blood proteins. The interaction of blood proteins such as human hemoglobin (Hb) with MTBE results in conformational and likely functional changes. The main mechanism for harmful effects of MTBE on Hb is through production of reactive oxygen species (ROS). In this regard, the present work was proposed to study the possible antioxidant potential of two dietary antioxidant agents, curcumin and caffeine, on the reduction of MTBE damage on Hb. Different spectroscopic methods including fluorescence, UV-Vis, circular dichroism, chemiluminescence, and molecular docking were used to study the interactions of curcumin and caffeine with Hb in the presence of MTBE. Our results showed caffeine could decrease the aggregation and ROS effects of MTBE on Hb. However, in the presence of curcumin the MTBE mediated aggregation of Hb was enhanced. These opposing effects of curcumin and caffeine as antioxidants were mainly contributed to the high iron chelating activity of curcumin. Thus, the complex formation between curcumin and heme further enhanced ROS production capability of MTBE.

Amigdalin (also known as laetrile or vitamin B17) a cyanogenic compounds, is widely distributed in plants, especially in the rosaceous plant seed, for example, apricot, peach, cherry, etc. It is a natural product that owns antitumor activity, less side effects and relatively low priced. Numerous studies have documented that amygdalin has antitussive and antiasthmatic effects, as well as an effects on the digestive system. Moreover, the pharmacological effects also include inhibition of renal interstitial fibrosis, prevention of pulmonary fibrosis, resistance to hyperoxia induced lung injury, immune suppression, immune regulation, antitumor, anti inflammatory and antiulcer. Different scientist tried to investigate the pharmacological activity, toxicity and antitumor effect in recent years, providing new insights for the development of new anticancer drugs, new targets searching and natural antitumor mechanism. In this paper summarizes published reports including different positive and negative utilization aspects of amigdalin and gives a better understanding of the known