دلاورشهباززاده

In recent years, the consumption of unhealthy, high-fat, and high-carbohydrate foods has led to an increase in obesity metabolic disorders, and cardiovascular diseases. This study aimed to investigate the effect of orally administering a ginger extract on serum lipid profiles, obesity factors, and cortisol and testosterone hormones in obese rats induced by a high-carbohydrate and high-fat diet. Twenty-one rats were divided into three groups. The first group was given a standard rat diet, the second group received a high-fat diet over 12 weeks, and the third group received a high-fat diet for 12 weeks with daily administration of ginger extract (100 mg) for six weeks. The intake of food and water, as well as weight gain, were measured. Additionally, plasma glucose, lipid profiles, and serum hormone levels were assessed biochemically. The results showed that the administration of ginger extract led to decreased food intake, weight ‎gain, glucose level, insulin level, and Homeostatic Model Assessment for Insulin Resistance (HOMA-IR) compared to obese ‎rats receiving a high-carbohydrate diet. Furthermore, cortisol levels remained unchanged in the ‎supplement-receiving group, and the increased testosterone in the test rat showed a significant ‎difference compared to the control rat. This study confirms that ginger extract could combat ‎obesity and related health problems by influencing metabolic processes and hormone levels, thus ‎helping to prevent the adverse effects of a high-carbohydrate diet.‎ ‎

Non‑small cell lung cancer (NSCLC) patients have a frequently poor prognosis and current therapies have large side effects. Numerous studies have been focused on the anti-cancer effects of melittin. Concerning the anti-cancer effects of melittin, this study aimed to evaluate melittin's anti-cancer effects on Calu-3 lung cancer cell line.

Cytotoxicity of melittin on Calu-3 and MRC5 cells was assessed using MTT assay. Real-Time PCR was used to determine expression of apoptotic and pro-apoptotic BAX, BCL2, and CASP3 genes. Furthermore, a wound healing assay was performed to compare the inhibition effects of melittin on the migration of interested cells.

IC50 values of melittin for Calu-3 and MRC5 were respectively 1.67μg/mL and 3.44μg/mL after 24h. Expression levels of BAX and CASP3 increased ‘2.82 and 4.31’ and ‘1.58 and 4.07’ fold in Calu-3 and MRC5, whereas BCL2 gene expression decreased 0.64 and 0.35-fold in the mentioned cell lines. It is demonstrated that cell migration is inhibited by melittin.

Melittin has an anti-cancer effect on Calu-3 lung cancer cell line. Furthermore, melittin induces apoptosis and inhibits cell migration of lung cancer cells. Melittin increases the apoptotic gene expression level and eventually causes the cells to move toward cell death. In addition, comparing the effects of melittin on Calu-3 cancer cells and MRC5 normal cells, it can be concluded that melittin has inhibitory and apoptotic effects on cancer cells at lower doses.

Skin cancer is the most common cancer and very high risk of metastasis of cancer. In recent years, proteins derived from natural sources such as poison animals are considered toxic. The aim of this study was purified Iranian cobra venom using chromatography & fractions obtained by examining toxicity and anti-tumor effect on skin cancer cell line.

B16F10 melanoma cells in RPMI 1640 medium with 10% FBS were cultured. Iranian cobra venom gel filtration chromatography and ion exchange was purified. Molecular weight fractions were analyzed by SDS-PAGE. The effect of anti-adhesion and cytotoxicity of the active fractions using trypan blue staining and MTT was measured on B16F10 cells were cultured and data were analyzed using student t-test.

Six fractions obtained of gel filtration chromatography. The isolated fractions were evaluated anti-adhesion effect and fraction F3 for further purification by ion exchange chromatography was nominated and 4 fractions were separated. The fraction with a molecular weight of about 47 kDa (F2) was nominated. Totally toxicity was calculated as 49% and 8% on skin cancer cell line and HEK-293, respectively (P≤ 0.05).

In this study, cytotoxic effect on skin cancer cells B16F10 showed that Iranian cobra venom have a protein with highly valued medicinal properties and could be an option for future research against skin cancer.

The fish's primary immune system in epidermis produces the mucosal layer as a surplus mechanism to cover the surface of the fish's body against infections. In addition to its physical functions such as reducing water friction and protecting against abrasion, it contains biologically active constituents, which is the first line of defense against pathogens. During the past years, the antimicrobial activity of various fish mucus has been studied. Due to the lack of information on sturgeon, in the present study, some properties of proteins in the epidermal mucus (Huso huso) as one of the native species of the southern coast of the Caspian Sea were evaluated. In this study, mucus was directly collected from fish surface in three different weight classes (0.4 , 4 and 40 kg) and some properties such as total protein concentration, chromatogram (RP-HPLC) and molecular mass profile of mucus proteins in three weight classes were compared. Antimicrobial activity of mucus was evaluated by disk diffusion, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The crude samples of different weights showed no significant difference in protein concentration (1.02 ± 0.03, 1.04 ± 0.01, 1.07 ± 0.02 µg/µl, respectively) but HPLC fraction No.4 in the third weight (40kg) showed a significant higher concentration in comparison to others (α=0.05). SDS-PAGE results revealed that the mucus samples contain peptides and proteins ranging from 5 kDa to 245 kDa with no considerable differences. All chromatograms showed similar numbers of fractions at similar retention time while the area percentage of fractions underwent significant differences in fractions 2, 4 and 9 (α=0.05). Disk diffusion showed the same antibacterial activity against A. hydrophila while group 1 (0.4 kg) had a significantly lower inhibition zone against S. iniae. All the samples possessed the same antimicrobial activity in MIC and MBC assays (MIC48=MBC48=25 µg). The current study revealed that all selected weight groups had the same properties in HPLC chromatogram and SDS-PAGE. Comparison of antimicrobial activity in MIC and MBC tests showed no significant difference between groups while in the disk diffusion test, there were significant differences in the antibacterial activity of weight groups exposed to S.iniaei, which could refer to the test competency. In conclusion, it can be claimed that in terms of protection (non-specific immunity), the first weight group (i.e. ≈ 0.4 kg) reached an evolutionary level as well as higher weights did (i.e. ≈ 4 and 40 kg).

Among Iranian venomous snakes, the most important groups causing envenomation are Naja Naja Oxiana, Echis,Vipera albicornuta, Vipera  latifii Mertens, pseudocerastes  peersicus and Vipera lebetina. Many researchers believe natural snake venom toxins are containing several pharmacologically active components  that could be of  potential therapeutic  value. In the  past, studies have  shown that  some neurotoxic fractions of snake  venom interfere with some infectious and non-infectious diseases such as cancer. we purified snake venom of V. lebetina by fast protein liquid chromatography (FPLC) using Sephacryl S-200 hr column. The fractions collected and evaluated by SDS-PAGE analysis.  The cytotoxicity effect of crude venom and fractions on Vero cells were demonstrated using 3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and adhesion assay. The aim of this study is to investigate the effects of Macrovipera  lebetina (one of the native snakes of many parts of iran) , in order to create an appropriate animal cell culture model to evaluate the effects of protein complexes on diseases such as viral infections.

Malaria is one of the most important parasitic diseases and one of the important health issues especially in tropical and subtropical areas of the world. .The importance of this disease is due to its high prevalence, significant mortality, as well as drug resistance and, side effects of current drugs in treatment. Venom is a complex mixture of active pharmaceutical ingredients. The purpose of this study was to investigate the anti-Plasmodium falciparum activity of the purified fraction of Iranian cobra snake venom by Real -time PCR.

After Lyophilized venom was purified for determining  inhibitory concentration 50% against Plasmodium falciparum 3D7 in vitro, in different dilution plates, different fractions obtained from Iranian cobra snake venom were adjacent to Plasmodium falciparum in the ring stage. The degree of parasitemia was determined. Finally, the effective fraction with antiplasmodial properties was identified.

The results showed that the most effective fraction had IC50 0.026 μg/mL on P. falciparum in vitro (p<0.001) .

The results were promising considering the anti-Plasmodium effect of the active fraction of Iranian cobra venom in the mentioned concentration and motivated the continuation of advanced research in this field.

Lung cancer is one of the most common cancer. Despite numerous therapies, chemotherapy is still the primary treatment . however Most patients, develop severe side effects. This has led to efforts to obtain new drugs from natural sources. The aim of this study was to investigate the cytotoxic effect of fractions obtained from Iranian cobra venom on A549 lung cancer cell line.

First, snake venom was purified by gel filtration method and the toxicity effect of the fractions obtained was evaluated by MTT assay on lung cancer cells and the selected fraction was purified by ion exchange method. The toxicity effect of the selected fraction was investigated on normal cells, the cell proliferation test was performed too and finally Real Time PCR test was performed on treated cells.

Fraction 6 of the gel filtration chromatography had the greatest cytotoxicity effect and was selected for further purification by ion exchange chromatography. Fraction 5 had the highest cytotoxicity effect. The toxicity of the selected fraction was significantly reduced on normal cells and the results of the cell proliferation test were consistent with the MTT results. Finally, Real Time PCR test was showed a significant increase in expression caspase 9 gene on treated cells.

Fraction 5 obtained from two stages of purification of Iranian cobra venom has cytotoxic effect on lung cancer cell line A549 and is a suitable option for drug studies of natural origin.

Biofilm is accumulation of microorganisms that are attached to a surface and they are coated with extracellular polymeric materials. Sometimes biofilm can be considered as a strategy that some microorganisms use to prevent harmful effects which are maintained in the natural environment and the host body. The purpose of this study was to evaluate antibiofilm effect of melittin peptide on clinical isolates of Pseudomonas aeruginosa isolated from nosocomial burn infections.

In this study, the rate of biofilm reduction and biodegradation kinetics of melittin on Pseudomonas aeruginosa biofilm were measured by microtiter plate crystal violet assay. The lack of growth in the MHA (Molar Hinton Agar) medium and decreasing in wavelength indicates the effect of melittin peptide on Pseudomonas aeruginosa biofilm.

The results showed that melittin was able to degrade the biofilm within 2 hours and killed the embedded bacteria. Melittin inhibited or eliminated all bacteria at the amount of 4 and 8 μg while at 50 μg biofilm layer was destroyed and all bacteria within the biofilm were killed after 24 and 48 h.

The results of this study are valuable while they increased the hope for treatment of pseudomonas associated infections in third degree burns patients.

One of the most dangerous endemic scorpions of Iran is Hemiscorpius lepturus that its venom is very toxic and known as responsible for most of scorpion sting dependent deaths in Iran. There are four Metalloproteinase inhibitors in the venom gland transcriptome of this scorpion (HLMetInhibit1-4). Metalloproteinase inhibitors are involved in some of the critical cells activity including the stability and surviving of ECM. This study aimed to investigate the structure and functions of a pair of these molecules and subsequently their cloning and expression for the first time.

Bioinformatics analysis and molecular docking on sequences of HLMetInhibit1 and HLMetInhibit2 performed (with the MG764541 and MG764542 NCBI accession numbers). The recombinant plasmids (pET-22b- HLMetInhibit 1,2) designed and synthesized, then transformed into the E. coli BL21 bacterial host. The colony-PCR and electrophoresis on one percent agarose gel performed and protein expression induced by different concentrations of IPTG at different times. Then SDS-PAGE and staining by coomassie brilliant blue and western blotting performed.

The highest binding affinity predicted for HLMetInhibit1 by molecular docking results. Gene cloning performed. Highest protein expression detected after 5 hours and with 1 mM concentration of IPTG. SDS-PAGE and western blotting confirmed the protein expression correctness for HLMetInhibit1 and HLMetInhibit2 with 26 and 17 KD of molecular weights.

Gene cloning and protein expression of Hemiscorpius lepturus metalloproteinase inhibitors 1 and 2 as the novel proteins that have not been studied so far, is approachable and it is hoped that after further in vitro and in vivo investigations, they will be the suitable candidates for research in the Various therapeutic fields.

Breast cancer is one of the most related cancer deaths among women. The scorpion venom consists of various peptides and proteins with cytotoxic, apoptotic and growth inhibitory effects on various cancers. Studying the cytotoxic effect Hemiscorpius lepturus scorpion venom on T47D breast cancer cell line was the main aim of this study.

The T47D cells were cultured as monolayer in RPMI1640 medium. The toxicity of the scorpion venom on T47D cell line was evaluated by MTT assay and the IC50 was calculated. The HEK293 cell line was used as control cell line.

The MTT assay results showed the time and dose-dependent inhibitory effect of H. lepturus venom on T47D cell line. However, such effect was not observed on HEK293 cell line in identical dose.

The achieved results indicate the anticancer effect of H. lepturus venom on T47D breast cancer cell line.

Vascular endothelial growth factor (VEGF) plays an important role in development of new blood vessel and angiogenesis. Human VEGF121 is smallest member of VEGF family. Production of active and correct form of VEGF is the most challenging issue.
Here we described a method for optimization of refolding of VEGF121 which was expressed in bacterial host. Enzyme linked immunosorbent assay and proliferation assay of human endothelial cells was performed to monitor refolding and functional assay of VEGF121.
Using described method; VEGF was in correct fold and detected by antibody in ELISA. Furthermore, VEGF stimulated proliferation of human endothelial cells in dose-dependent manner.
Refolded VEGF has potential for stem cell differentiation.

Cancer is a complex disease characterized by uncontrolled cell proliferation. It is a main global public health problem. Chemotherapy is the dominant method in the treatment of cancer and patients often show tumors that become resistant to the drug, leading to serious side effects and death. Brain cancer is about 31% of all cancers detected among children. Scorpion venoms are a source of useful biologically active compounds with anti-tumor effects. The purpose of this research, was purification of the fractions from the venom of Iranian scorpion, Hemiscorpius lepturus by gel filtration chromatography and evaluation of its anticancer activity on brain cancer cell line .
Scorpion venom was prepaired and after concentration determination, the venom fractions were purified by gel filtration chromatography using sephadex G50 and deried in a freeze dryer system. The molecular weight ranges of the fractions were detected in SDS-PAGE. The anticancer effect of each fraction was examined using different protein concentrations on U-87 MG cell line.
According to data obtained from gel filtration, seven FPLC fractions were collected. As the fraction No.4 was contained peptide fraction used for evaluation of anticancer activity. The anticancer effect of the peptide fraction on brain cancer cells was 47% in 0.31µg. No toxicity was observed on with fibroblast cells at this concentration.
Iranian scoprion venom had significant activity on brain cancer cell lines and no toxicity was shown on control cells. The use of natural compounds as potential anti-cancer agents that are derived from venomous animals could be assisted to fighting against cancer. This study is the first report regarding to existence of an anticancer fraction that is effective on glioblastoma cancer cell line derived fron the venon of Iranian scrorpion, H. lepturus.

Numerous proteins, peptides, and chemical agents in the venom of venomous marine animals are potentially useful biologically active molecules with pharmacological properties. The main goal of this research was to study the cytotoxic effects isolated crude venom from the Persian Gulf sea anemone, Stichodactyla haddoni against Breast, and Human embryonic kidney cell lines in vitro through using MTT assay. Samples of S. haddoni were collected from coastal waters of Lark island Persian Gulf the south of Iran. The extraction of Aqueous from tentacle was performed. The cell line was cultured in complete tissue culture medium. These cells were exposing in presence of different Serial dilution 100 to 0.78 μg of venom extract in duration time 24 hour. Analysis of statistical for cytotoxic activity of crude venom on cell lines this study showed that activity was similar together in almost all doses(Pvalue

Cancer is a life threatening disease determined by uncontrolled differentiation and proliferation of cells. The cells escape apoptosis would be led to metastasis. Chemotherapy is the conventional method in treatment of cancer but the tumor often will be resistant to chemotherapy regimen leading to crucial side effects. Prevalence of lung cancer is 19.4% of all cancers. Many bio-toxins are useful source for biologically active compounds with anti-tumor activity. This study was aimed to finding an anti-cancer agent from the venom of Iranian snake viper, Macrovipera lebetina. The venom fractionated and anti-adhesion activity was examined. Candidate fraction selected and subjected to anion exchange chromatography to further fractionation. Subsequently, anti-adhesion and toxic activity determined on lung cancer line.
Mateials and Viper venom prepared and separated by gel filtration chromatography using sephacryl S200.The concentration of fractions determined and anti-adhesion effect of the fractions evaluated on TC-1 cell line. Then, the active fraction was purified by anion exchange chromatography using MonoQ resin. Anti-adhesion activity and toxicity of resultant fraction determined on the cancer cells again. Toxicity of candidate fraction controlled on fibroblast cell line.
Based on results six fractions were collected. Fraction 5 was active on cell lines and selected for anion exchange chromatography .Four anionic and two cationic fractions showed in the chromatogram and collected for anticancer activity. Fraction 3 has the most anti-adhesion activity. Totally anti-adhesion and toxicity on cancer cell line calculated as 40%. This fraction showed about 5% toxicity on fibroblasts.
An anti-adhesion fraction was detected in the venom of Iranian viper snake, Macrovipera lebetina. Toxicity of candidate fraction on cancer cells was less than anti-adhesion activity. Less toxicity was showed on fibroblast cells. Natural anti-cancer compounds derived from venomous animals can help fighting against cancer. It is the first report of an anti-adhesion fraction from an Iranian Viper with less toxicity on normal cells. Protein sequencing and in vivo study in mouse model of lung cancer would be suggested.

For many years, antibiotic resistant infections including septicemia, burn infection, and peritonitis have been reported from many hospitals and still no antibiotic guarantees comprehensive treatment. Antimicrobial peptides are one of suggested solutions to overcome the problem. The goal of this study was tracing for an antimicrobial peptide in the venom of Iranian cobra snake, Naja naja oxiana and evaluation of its toxicity on human Red Blood Cells as well.
Snake venom was firstly fractionated using Gel filtration chromatography. The resulted fractions controlled for antibacterial activity against E. coli and Staphylococcus aureus. The fractions responsible for antibacterial activity were fractionated by Reverse Phase- HPLC. All major peaks were collected and controlled for activity against E. coli. Positive fractions selected to examination of activity against Pseudomonas aeruginosa (ATCC27853), Acinetobacter baumannii (ATCCBAA747), and Staphylococcus aureus (ATCC29213). Toxicity of candidate antibacterial fraction was evaluated by in vitro hemolysis assay.
The weight of proteins and peptides approximately ranged from 7KDa to 170 KDa. According to results, 8 fractions were obtained gel filtration chromatography. Among 8 fractions only 3 fractions were peptide and had the most area percent. Three fractions, F6, F7, and F8, selected for determination of MIC against E. coli and S. aureus. Two fractions had antibacterial activity at 50µg for both of examined bacteria. As the amount of fraction F7 was greater than F6, it was candidated for further fractionation by HPLC. 12 peaks were eluted and collected from column. Among the fractions, F4, F9, F10, and F12 were more effective against E. coli. Fraction 12 had the most activity as compared to the others. The weight of F12 approximately estimated as an 8KDa peptide. The MIC for F12 against E. coli, Staphylococcus aureus, and Pseudomonas aeruginosa were determined at 6.25, 25, and 50µg respectively.
We could find a bactericidal peptide. It is the first report for antibacterial activity of Iranian cobra snake venom. This peptide did not show any direct or indirect hemolytic activity. Natural compounds like antimicrobial peptides derived from venomous animals could be an effective solution to deal with multidrug resistant pathogen.

Breast cancer is the leading cause of cancer-related death in women. Many biotoxins like snake venom as a source of useful biologically active compounds are potential anti-cancer agents. Venom is a complex mixture of active pharmaceutical ingredients. The purpose of this study was investigation of anticancer activity of the fractions obtained from the venom of Iranian cobra,Naja naja oxiana, by gelfiltration and anion exchange chromatography on breast cancer cell line.
Breast cancer cell line (4T1) was cultured in RPMI with 10% fetal bovine serum. Iranian cobra venom purified by gel filtration and anion exchange chromatography. The concentrations of these fractions were determined by BCA method and their molecular weight measured by SDS-PAGE. Cytotoxicity of the fractions on cancer and normal cell lines was evaluated with MTT assay.
Based on the results obtained from gel filtration chromatography, 6 fractions were separated. Fraction F3 had a protein with molecular weight of 50 kDa that is equivalent to the molecular weight of disinterring. Therefore, fraction F3 selected for further purification by anion exchange chromatography. Four fractions were isolated by anion exchange chromatography. Fraction F2 had anticancer activity by MTT and Anti-adhesion assays and determined as active anti-cancer fraction. Molecular weight of fraction F2 was 47 kDa. The results of MTT assay showed 45 and 8 percent cytotoxicity on 4T1 and normal cell lines(HEK293) respectively.
The active fraction of Iranian Cobra venom had cytotoxic effects on breast cancer cells with very low toxicity on normal cells. According to results, this fraction had anti-adhesion and cytotoxic activity can be a good candidate for more purification and characterization.

Infections caused by Pseudomonas aeruginosa have been reported from many hospitals، and according to its opportunistic nature، Pseudomonas aeruginosa invades to immunocompromised patients and controlling the following infection is so hard. Antibiotics misusage beside complex mutation mechanisms in pseudomonas genus led to resistance against many antibiotics. During the past decade، tracing for natural antimicrobial peptide is more considered. Among them، melittin has been extracted from honey bee venom and its antibacterial activity is being examined. The main goal of this study was to evaluate antibacterial activity of melittin against clinical isolates of Pseudomonas aeruginosa. 48 Clinical strains were present from previous study. Honey bee venom was prepared using electrical stimulation and the quality of venom was confirmed by SDS-PAGE. Melittin was isolated from the venom using a linear gradient of acetonitrile and C18 column by Reverse Phase-High Performance Chromatography (RP-HPLC) Technique. Minimal Inhibition and Bactericidal concentration of melittin were examined on clinical isolates of Pseudomonas aeruginosa. Honey bee venom consisted of twenty distinct fractions in which melittin was the major one. MIC50 for melittin against all; mucoid، and non-mucoid sensitive isolates، were 12. 5، 6. 25، and 12. 5 μg respectively. Among mucoid and non-mucoid strains، 94. 73 and 86. 2 % were sensitive to melittin respectively. MBC50 for melittin against total، mucoid and non-mucoid isolates were 50، 37. 5، and 50 μg respectively. Totally، 89. 58 % were sensitive to melittin and 10. 41 % were not inhibited. The results obtained from MIC and MBC tests showed that melittin had significant inhibitory and bactericidal effect on clinical isolates of pseudomonas aeruginosa. According to the results، the less amount of melittin can suppress the growth of mucoid strain of pseudomonas. These strains due to existence of polysaccharide layers around bacteria have less permeability to antibiotics and expose more resistance. This study would be of great value to the treatment of cystic fibrosis patients with pseudomonas infections.

Prostate cancer is one of the most common cancers diagnosed in developed countries. Many studies have confirmed that the nm23 gene suppresses metastasis in different types of cancers. The effects of imatinib as the first member of tyrosine kinases inhibitors were showed in research and treatment of solid tumours. The aim of the current study was to investigate the effect of imatinib on cell viability and Suppressor metastasis nm23 gene expression in prostate cancer cell line. In this study, Prostate cancer (PC-3) human cell line was treated with various concentrations of imatinib for 48hours. Cell viability was assessed using MTT assay and the IC50 value was determined. We extracted RNA molecules by using RNX solution, after which cDNA was synthesized. The precise primers for the nm23 and GAPDH genes were designed by specific software. Then, the quantity of nm23 compared to GAPDH gene (reference gene) was analyzed using real-time PCR. Imatinib exerted an inhibitory effect on the viability of metastatic PC-3 cells. The calculated nm23/GAPDH gene expression ratio were 1.62±0.02 (p<0.01) in 21/33mM concentration of imatinib at 48h. The result of this study was shown that imatinib can inhibit metastasis by up regulating nm23 gene expression in Prostate cancer PC-3 cell line.

Background and Hirudin is a 65-66 amino acids polypeptide which is secreted as ananticoagulant compound from salivary glands of medical leech. This drug is a very potent inhibitor of thrombin and is so effective for arterial and venous thrombosis prevention. Therefore, it can compete with heparin. The aim of this study was to add a pelB signal peptide to pET-22b plasmid and to investigate the expression of recombinant hirudin in E.coli. At first, the 66 nucleotic sequence of hirudin's gene was obtained and entered to a powerful vector (pET-22b). N_terminal His-tag was used for protein purification. We inserted pelB signal sequence at the begining of the gene to secrete protein to periplasmic region and culture medium. The expression of the target protein by origami (DE3) strain was then measured. Finally, the target protein was measured in periplasmic region, cytoplasm and culture medium. The results showed that more protein was expressed in the periplasmic space and a small amount of protein secreted into the culture medium. Using the vectors capable of transferring the proteins to the periplasmic region, that is very favorable space for the formation of disulfide bonds and properly folding of the target protein, could decrease the production of inclusion bodies and increase the production of soluble andactive proteins.

Bacterial peritonitis is one of the nosocomial infections that is due to direct invasion of bacteria to peritoneal membrane. Resistance to antibiotic is of great significance in this disease and could be led to morbidity and mortality of patients. During the past decade, tracing for natural antimicrobial peptide is more considered. Among them, melittin has been extracted from honey bee venom and its antibacterial activity is being examined. The main goal of this study was isolation of melittin from honey bee venom and evaluation of its antibacterial activity against the agents of bacterial peritonitis. Honey bee venom prepared using electrical stimulation and the quality of venom confirmed by SDS-PAGE. Melittin isolated from the venom using a linear gradient of acetonitrile and C18 column by Reverse Phase-High Performance Chromatography (RP-HPLC). Minimal Inhibition and Bactericidal concentration for melittin examined on Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. Honey bee venom composed of twenty distinct fraction in which melittin was the major one. Melittin inhibited Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa growth at 0.39, 6.25, and 12.5 μg and was bactericide at 1.56, 25, and >50 μg respectively. Melittin specifically invade the corresponding bacteria and induce significant inhibitory and bactericidal activity against the main agents of bacterial peritonitis. Complementary studies in animal model would be overcome bacterial drug resistance issue specifically in bacterial peritonitis.

Cervical cancer is the second prevalent cancer in developing countries and the sixth prevalent cancer in USA. Since conventional treatment methods are associated with detrimental side effects، searching for new drugs using natural ingredients is very important. Previous studies have shown that melittin (main component of honey bee venom) has anticancer properties along with the effect on cell membrane and activation of apoptosis. In this study، inhibitory effects of melittin on the viability and proliferation of cervical cancer cell line (HeLa) was investigated.

Melittin was purified from honeybee venom using reversed-phase HPLC method. Then، biological activity of melittin was examined by hemolytic activity analysis on the red blood cells. In order to investigate whether melittin inhibits proliferation of HeLa cell، MTT assay was performed. HeLa cells were plated in a 96-well plate and treated with serially diluted concentrations of melittin for 12 and 24 hours. The viability of the cells was measured via MTT assay at 540nm.

Melittin showed a strong hemolytic activity (HD50=0. 5 µg/ml) which can be reduced by FBS (HD50=2 µg/ml). Results of MTT assay indicated that melittin shows cytotoxic effect on cervical cancer cells with IC50 = 1. 2 ug/ml at 12h incubation period.

In this study، biological activity of melittin and inhibitory effect of FBS on hemolysis were determined via hemolytic activity analysis. MTT assay indicated that melittin induced cytotoxic effects in a dose dependent manner on cervical cancer cells and it also revealed dependence on incubation time as well.

Ritalin has high tendency to be abused. It has been the main indication to control attention deficit hyperactivity disorder. The college students may seek for it to improve their memory، decrease the need for sleep (especially during exams)، which at least partially، can be related to serotonergic system. Therefore، it seems worthy to evaluate the effect of Ritalin intake on mature brain. There are many studies on Ritalin effect on developing brain، but only few studies on adults are available. This study was undertaken to find Ritalin effect on serotonin transporter (SERT) density in medial frontal cortex (MFC) of mature rat. Thirty male Wistar rats were used in the study. Rats were assigned into five groups (n = 6 per group): one control، two Ritalin and two vehicle groups. Twelve rats received Ritalin (20 mg/kg/twice a day) orally for eleven continuous days. After one week of withdrawal and another two weeks of rest، in order to evaluate short-term effects of Ritalin، six rats were sacrificed. Another six rats were studied to detect the long-term effects of Ritalin; therefore، they were sacrificed 12 weeks after the previous group. The immunohistochemistry was performed to evaluate the results. Immunohistochemistry studies showed a higher density of SERT in both 2 and 12 weeks after withdrawal from Ritalin intake in MFC of rat and there was no significant difference between these two groups. Our findings demonstrated both short- and long-term effects of Ritalin on frontal serotonergic system after withdrawal period.

Identification of venomous species of Persian Gulf cone snails and characterization of venom composition and their features is so important from the point of danger for underwater divers as well as medical importance. In this research, the specimens of Conus textile were collected off Larak Island from depth of 7 m. The collected samples were transferred to laboratory alive and were stored at -700C. The venom´s ducts were separated and homogenized with deionized water. The mixture centrifuged at 13000 rpm for 15 minutes at 4 ̊C. Supernatant was considered as extracted venom and stored at -20 C after lyophylization. The entity of venom determined by SDS-PAGE and RP-HPLC methods used to investigate the extracted venom and system to determine protein pattern. The results obtained with SDS-PAGE showed that proteins and peptides of venom were ranged between 6 to 250KD. Chromatogram of the venom demonstrated more than 44 large and small fractions.