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Background: COVID-19 now is a serious concern for the world healthcare system. This study aimed to investigate possible therapeutic effect of colchicine and phenolic monoterpenes accompanied by standard care of treatment (SCT) in patients diagnosed with COVID-19. Methods: In this randomized controlled parallel clinical trial, a total number of 179 (of 200) patients with confirmed COVID-19 were enrolled according to the inclusion and exclusion criteria. The patients were allocated by simple randomization method into two groups control (receiving SCT with 71 patients) and intervention (receiving SCT plus colchicine and phenolic monoterpenes with 107 patients). The mortality ratio during hospitalization as well as a 2-week follow-up, ICU admission rate, and hospitalization duration were assessed as main outcomes. Results: The mortality ratio was 0.9% (1/108) and 8.45% (6/71) in the intervention and the control groups (p-value = 0.035) respectively, these ratios after a 14-day follow-up were 1.85% (2/108), and 9.85 (7/71) respectively (p-value = 0.031). Also, the ICU admission was significantly lower (p-value = 0.006) in the intervention group 2/108 (1.85%) compared with controls 10/71 (14.08%). Moreover, the duration of hospitalization followed a similar pattern to ICU admission with 4.17 ± 1.34 vs. 6.39 ± 2.59 days in the intervention and control groups respectively (p-value< 0.001). Furthermore, no significant side effect was found between the groups. Conclusion: According to the results, the combination of colchicine plus phenolic monoterpenes could be an additive treatment for the SCT. The authors strongly recommend further trials on this combination with other SCTs.

ObjectiveBreast cancer is one of the major causes of mortality among women. Due to many side effects of the existingchemotherapeutic agents, the research of anti-cancer drugs, including natural products, is still a big challenge. Here,we investigated the effects of colchicine on apoptosis of two breast cancer cell lines ( human MCF-7 and mouse 4T1).Materials and MethodsIn this experimental study, we evaluated the apoptotic effects of colchicine on (MCF-7) and(4T1), as well as a human cancer-associated fibroblast cell line as a control group. Extraction and chromatographictechniques were applied to isolate colchicine from Colchicum autumnale L. To compare the isolated colchicine withpure standard colchicine, we used the H-NMR technique. The methyl thiazolyl tetrazolium (MTT) assay, quantitativereverse transcriptase-polymerase chain reaction, Western blotting and annexin V/PI staining were used to evaluate theapoptotic effects of the isolated and standard colchicine.ResultsSimilar to standard colchicine, the isolated colchicine inhibited cell proliferation significantly in cancer celllines. Colchine inhibited proliferation and induced apoptosis on a dose-dependent manner. The medicine modified theexpression of genes-related to apoptosis by up-regulation of P53 ,BAX, CASPASE-3, -9 and down-regulation of BCL-2gene, which led to an increase in the BAX/BCL-2 ratio.ConclusionWe showed that isolated colchicine from Colchicum autumnale and pure standard colchicines modulatethe expression levels of several genes and therefore exerting their anticancer effects on both human (MCF-7) andmouse (4T1) breast cancer cells. Based on these results, we suggest that colchicine can be a potential candidate forprevention and treatment of breast cancer.

Introduction: Further development of magnetic-based detection techniques could be of significant use in increasing the sensitivity of detection and quantification of hepatitis B virus (HBV) infection. The present work addresses the fabrication and characterization of a new bio-nano composite based on the immobilization of goat anti-HBsAg antibody on modified core-shell magnetic nanoparticles (NPs) by (3-aminopropyl) triethoxysilane (APTES), named Fe3O4@SiO2/NH2, and magnetic NPs modified by chitosan (Fe3O4@CS). Methods: At the first step, Fe3O4 was modified with the silica and APTES (Fe3O4@SiO2/NH2) and chitosan (Fe3O4@CS) separately. The goat anti-HBsAg antibody was activated by two different protocols: Sodium periodate and EDC-NHS. Then the resulted composites were conjugated with activated goat anti-HBsAg IgG. An external magnet collected Bio-super magnetic NPs (BSMNPs) and the remained solution was analyzed by the Bradford method to check the amount of attached antibody to the surface of BSMNPs. Results: The findings indicated that activation of antibodies by sodium periodate method 15-17 µg antibody immobilized on 1 mg of super magnetic nanoparticles (SMNPs). However, in the EDC-NHS method, 8-10 µg of antibody was conjugated with 1 mg of SMNPs. The resulting bio-magnetic NPs were applied for interaction with the HBsAg target using enzyme-linked immunosorbent assay (ELISA). About 1 µg antigen attached to 1 mg SMNPs, which demonstrated that the fabricated materials are applicable in the detection scope of HBsAg. Conclusion: In the present study, we developed new antibody-conjugated magnetic NPs for the detection of HBsAg using an efficient conjugation strategy. The results demonstrated that the binding capacity of Fe3O4@SiO2/NH2 was comparable with commercially available products. Our designed method for conjugating anti-HBsAg antibody to a magnetic nanoparticle opens the way to produce a high capacity of magnetic NPs.

AIM: To assess the effectiveness, safety, and cost-effectiveness of the Argus II in treatment of the retinitis pigmentosa (RP) patients. METHODS: The ProQuest, Web of Science, EMBASE, MEDLINE (via PubMed) were searched using combinations of the keywords of Argus, safety, effectiveness, bionic eye, retinal prosthesis, and RP through March 2018. The retrieved records were screened and then assessed for eligibility. RESULTS: Totally 926 records were retrieved from the searched databases and finally 12 studies included. The RP patients showed improvements in visual function after receiving the prosthesis, compared to the time before the prosthesis or the time it was off. This was measured by square localization, direction of motion, and grating visual acuity tests. No major adverse effect was reported for the Argus II prosthesis itself and/or the surgery to implement it, but the most frequently reported items were hypotony, and conjunctival dehiscence. The incremental cost-effectiveness ratio (ICER) was calculated to be €14603 per quality-adjusted life year (QALY) in UK and $207 616 per QALY in Canada. CONCLUSION: The available evidence shows that the Argus II prosthesis in RP patients is effective in improvement of their visual function. Some minor adverse effects are reported for the prosthesis. The cost-effectiveness studies show that the technology is cost-effective only at high levels of willingness-to-pay.

Over recent years, nosocomial infections, and the morbidity and mortality associated with pathogenic bacteria have dramatically increased due to antibiotic resistance and imposed significant burdens on the global health system. Critical shortage of effective therapeutics against multidrug-resistant bacteria highlights the need for development of novel antibiotics. Actinobacteria are well-known sources of natural bioactive compounds, especially antibiotics. Nearly two-thirds of the antibiotics on the market have actinobacterial origins. Endophytic actinobacteria residing within plants contribute to the plant growth and survival by producing plethora of secondary metabolites. Therefore, isolation, cultivation, and identification of new strains, as well as their potential to produce antimicrobial compounds, are of great importance. Lack of published research in this field highlights the importance of this review in Iran. The aim of this review was to present the latest methods for the isolation and identification of endophytic actinobacteria and introducing relevant databases. We also studied the most recent isolated strains, chemical structure of 51 newly identified secondary metabolites, and their potential medical-pharmaceutical applications. This study revealed that endophytic actinobacteria are prolific sources of bioactive secondary metabolites with high levels of structural diversity and potent pharmaceutical and medicinal applications.

Background: Atherosclerosis is a chronic inflammation that interferes with blood arteries functions due to the accumulation of low density lipids and cholesterol. Objective: To investigate the effect of aqueous extract and saponin fraction of Tribulus terrestris L. (TT) on the proteome and expression of intracellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and E-selectin in the human umbilical vein endothelial cells (HUVEC) and human bone marrow endothelial cell (HBMEC) lines. Methods: Two cell lines were cultured and induced with lipopolysaccharide (LPS). The primed cells were then treated with aqueous extract and saponin fraction of TT. The protein profile of the endothelial cells was assessed under normal and LPS-induced conditions using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and 2D gel electrophoresis (2-DE). The levels of VCAM-1, ICAM-1, and E-selectin were estimated by use of western blotting. Results: LPS-induced HUVECs and HBMECs were shown to significantly increase the expression of ICAM-1, VCAM-1, and E-selectin in comparison to control groups. Our findings revealed that TT extract resulted in significantly more reduced levels of proteom (80 spots) as well as all the three mentioned proteins compared with the effect of saponin fraction alone. Conclusion: TT extract and its saponin fraction exerted anti-inflammatory effects on HUVEC and HBMEC lines and reduced the expression of ICAM-1, VCAM-1, and E-selectin. However, the anti-inflammatory effect of aqueous extract was greater than that of saponin fraction. Therefore, TT could be considered as a potential candidate for the treatment or prevention of atherosclerosis.

Background: Breast cancer is the most common cause of cancer-related death in women worldwide. Therefore, there is an urget need to identify and develop therapeutic strategies against this deadly disease. This study is the first to investigate the effects of Hemolymph Serum of Potamon persicum Crab (HSPPC) on MCF-7 and MDA-231 breast cancer cell lines. Materials and Methods: LDH and MTT assays were performed on MCF-7 and MDA-231 breast cancer cell lines as well as human umbilical vein endothelial cells (HUVEC) to determine the cytotoxic and antiproliferative activity of the HSPPC at different concentrations. Further, the apoptosis inducing action of the hemolymph serum was determined by TUNEL (terminal deoxynucleotidyl transferasemediated dUTP nick end labeling) and cell death assay. Results: The IC50 values of HSPPC for MCF-7 and MDA-231 cell lines were 960±0.369 and 850±1.422 μg/mL, respectively. The growth of both MCF-7 and MDA-231 cell lines were significantly (P

Background: Chondroitin sulfates (CS) is an important glycosaminoglycan’s which is found in sturgeon fishes which are present in cartilage of these fishes. This compounds, in addition to application in food and cosmetic industries, they have clinical significant in arthritis treatment and wound healing. Materials and Methods: In present study, the chondroitin sulfate extraction was performed by using pepsin and trypsin digestions with 12 and 18 h interval times followed by cationic salt, cetylprydinium chloride (CPC) treatment. MTT assay at different incubation time (24, 48 and 72 h) was used in order to study of Chondroitin sulfate effect on induction of proliferation of fibroblasts isolated from human skin. In order to determine the proper time and enzymatic digestion, after temperature treatment of samples using pepsin and trypsin enzymes, the SDS-PAGE electrophoresis was used. And the FT-IR analysis was performed to characterize the type of CS. Results: The maximum yield of enzymatic digestion was observed for trypsin digestion at 18h, according to electrophoresis patterns. the amount of CS yield was estimated at 4.76% in this situation. FT-IR analysis revealed that, the CS belongs to the C4S type. Chondroitin sulfates extracting from cartilage showed concentration-dependent incremental impact on fibroblast cell proliferation so that the dose-response results have shown 167% proliferation at 10 µg/ml dose of CS compared to the control in fibroblast cells. Conclusion: Extracted CS has a positive stimulatory effect on fibroblast cells proliferation in a dose dependent manner. So it can be used for wound healing and some fibroblast related diseases.

In the physiological study of stem remobilization of assimilates during grain filling, mutant genetic materials are a valuable tool. Two advanced mutant lines of bread wheat (T-67-60 and T-65-7-1) along with their wild type (Tabasi Cv.) were planted at two moisture conditions (normal and 30-40% of field capacity) as a factorial experiment based on a completely randomized design with three replications. Drought treatment initiated at full heading stage (Zadoks 60) and sampling done at 5 steps (0, 7, 14, 21 and 28 days after anthesis). Based on the results remobilization of stem and its efficiency in the mutant lines was significantly more than wild type, under terminal drought stress. The reasons for this were higher capability of the sink and early senescence in the mutant lines compared to the wild type. In consequence of changes in chlorophyll content and relative gene expression of key photosynthetic Rubisco (Rubisco large and small subunits and Rubisco activase), in the genotypes during grain filling seems degradation of chloroplasts and photosynthetic system occurred caused by senescence drought stress-induced in the mutant lines more strongly than in the wild type, and because of senescence is stimulating remobilization of assimilates from stem to grain, remobilization of stem and its efficiency in mutant lines were more than wild type. Also, wild type further with continuation of current photosynthesis in the leaves has acted for the grain filling.

Context: Ellagic acid (EA) is a natural phenol antioxidant with various therapeutic activities. However, the efficacy of EA has not been examined in neuropathologic conditions. Objective: In vivo neuroprotective effects of EA on cuprizone (cup)-induced demyelination were evaluated. Material and methods: C57BL/6 J mice were fed with chow containing 0.2% cup for 4 weeks to induce oligodendrocytes (OLGs) depletion predominantly in the corpus callosum (CC). EA was administered at different doses (40 or 80 mg/kg body weight/day/i.p.) from the first day of cup diet. Oligodendrocytes apoptosis [TUNEL assay and myelin oligodendrocyte glycoprotein (MOG+)/caspase-3+ cells), gliosis (H&E staining, glial fibrillary acidic protein (GFAP+) and macrophage-3 (Mac-3+) cells) and inflammatory markers (interleukin 17 (IL-17), interleukin 11 (IL-11) and stromal cell-derived factor 1 α (SDF-1α) or CXCL12] during cup intoxication were examined. Results: High dose of EA (EA-80) increased mature oligodendrocytes population (MOG+ cells, p

Background: Formation of secondary structure such as DNA hairpins or loops may influence molecular genetics methods and PCR based approaches necessary for genetic engineering, in addition to gene regulation. Materials and Methods: A polymerase chain reaction with splice overlap extension (SOE-PCR) was used to create fully synthetic 1F5 chimeric anti-CD20 heavy- and light-chain genes. The chimeric genes were cloned into the pCR-Blunt II-TOPO vector following by cloning into the pBudCE4.1 expression vector. Prediction of secondary structure was performed with the Vienna RNAfold webserver. PCR and sequencing across the predicted secondary structure of chimeric 1F5 heavy-chain gene was performed with multiple protocols for standard and GC-rich templates. Results: In our attempt to design vectors aimed to generate mouse-human chimeric antibody against CD20 (1F5), we found that the coding sequence of 1F5 chimeric heavy-chain gene constructed by SOE-PCR was resistant to polymerase during both PCR and sequencing reactions. Furthermore, we were also unable to analysis some positive transformants by restriction enzyme digestion. Encountering such difficulties to identify the cloned anti-CD20 chimeric heavy-chain gene, we found that the chimeric heavy-chain sequence is highly GC-rich and predicted to form a stable secondary structure. Conclusion: In conclusion, for the first time, we reported several difficulties with production of therapeutic chimeric 1F5 anti-CD20 antibody due to a predicted hairpin cluster correlates with barriers to PCR, sequencing and possibly restriction analysis. Our findings provide a probable note for researchers experiencing technical difficulties with construction of chimeric anti-CD20 antibody 1F5 gene vectors and also with other genes and molecular biology techniques requiring PCR-based method or restriction enzyme analysis.

Angiogenesis has essential role in growth and metastasis of tumors. Development of therapies aimed to suppress angiogenesis using medicinal plants is one of the effective approaches for prevention/treatment of cancer. The current study was performed to investigate in vitro anti-angiogenic effect of Teucrium Polium (TP) extract and its fractions. The aerial part of Teucrium Polium was powdered and extracted with 50% ethanol. The extract was fractionated in to aqueous (AQ), n-butanol (BU), ethyl acetate (EA) and n-hexane (HE) fractions. Anti-angiogenic effect of TP was examined on human umbilical vein endothelial cells (HUVECs) in three-dimensional collagen matrix. The endothelial cells form capillary-like branches that can be visualized using phase contrast microscope and the number of tube-like structures can be quantified as a measure of in vitro angiogenesis. Furthermore, anti-proliferative and vascular endothelial growth factor(VEGF )suppressive effect of TP as important factors in the process of angiogenesis were assessed using3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT)and quantitative ELISA, respectively. Based on our findings, among the TP fractions, EA fraction showed the highest inhibitory activity on angiogenesis. This fraction with IC50: 68 µg/mL, inhibited angiogenesis at 60 µg/mL. The crude extract and other fractions of TP inhibited angiogenesis in a dose-dependent manner at doses higher concentrations than EA fraction, significantly.TP extract and EA fraction were able to inhibit proliferation of HUVEC and inhibited VEGF secretion in a dose dependent manner. The ethyl acetate fraction at 60 µg/ml inhibited VEGF secretion perfectly. Our data indicated that ethyl acetate fraction of Teucrium Polium could be a potential candidate for the prevention of angiogenesis in cancer and other related disorders. However, this suggestion needs more quantitative and in vivo investigations for confirmation.

Objective(s):Multi-epitopic protein vaccines and direction of vaccine delivery to dendritic cells (DCs) are promising approaches for enhancing immune responses against mutable pathogens. Escherichia coli is current host for expression of recombinant proteins, and it is important to optimize expression condition. The aim of this study was the optimization of multi-epitopic HIV-1 tat/pol/gag/env recombinant protein (HIVtop4) expression by E. coli and conjugation of purified protein to anti DEC-205 monoclonal antibody as candidate vaccine. Materials and Methods: In this study, expression was induced in BL21 (DE3) E. coli cells by optimization of induction condition, post induction incubation time, temperature and culture medium formula. Some culture mediums were used for cell culture, and isopropyl-beta-D-thiogalactopyranoside was used for induction of expression. Protein was purified by Ni-NTA column chromatography and confirmed against anti-His antibody in western-blotting. To exploit DCs properties for immunization purposes, recombinant protein chemically coupled to αDEC-205 monoclonal antibody and confirmed against anti-His antibody in western-blotting. Results: The optimum condition for expression was 1 mM IPTG during 4 hr cultures in 2XYT medium, and final protein produced in soluble form. Conjugation of purified protein to αDEC-205 antibody resulted in smears of protein: antibodies conjugate in different molecular weights. [AGA1] . Conclusion: The best cultivation condition for production of HIVtop4 protein is induction by 1 mM IPTG during 4 hr in 2XYT medium. [AGA2] The final concentration of purified protein was 500 µg/ml.

The follicular fluid (FF) is produced during folliculogenesis and contains a variety of proteins that play important roles in follicle development and oocyte maturation. Age-related infertility is usually considered as a problem that can be solved by assisted reproduction technology. Therefore, the identification of novel biomarkers that are linked to reproductive aging is the subject of this study. FF was obtained from healthy younger (20–32 years old) and older (38–42 years old) women undergoing intracytoplasmic sperm injection (ICSI) due to male factor infertility. The FF was analyzed by two-dimensional gel electrophoresis (2-DE). The power of two-dimensional gel electrophoresis and the identification of proteins were exploited using matrix-assisted laser desorption-ionization time-of-flight/time-of-flight (MALDI-TOF-TOF) mass spectrometry. Twenty three protein spots showed reproducible and significant changes in the aged compared to the young group. Of these, 19 protein spots could be identified using MALDI-TOF-TOF-MS. As a result of MASCOT search, five unique downregulated proteins were identified in the older group. These were identified as serotransferrin, hemopexin precursor, complement C3, C4 and kininogen. A number of protein markers were found that may help develop diagnostic methods of infertility.

Introduction & Objective: Ficin is a member of plant cystein proteases that is abundant in fig. This enzyme has many pharmacological and industrial uses. In the present study, the enzyme was purified by a simple procedure and its proteolytic activity was assayed on several plant and animal proteins. Materials & Methods: Ficin was extracted from unripe fig, precipitated by ammonium sulfate and purified using ion-exchange chromatography on a Carboxymethyl Sepharose column. Proteolytic activities of the purified enzyme were determined in 4 buffering conditions on casein, alpha lactalbumin, beta lactoglobulin and gelatin proteins. Results: Purified enzymes include two bands with molecular mass of 24 and 26 KDa. Results of proteolytic activity showed that ficin can digest casein. It has moderate hydrolytic activity on beta lactoglobulin and gelatin but ficin can not hydrolyze alpha lactalbumin. Conclusion: It seems ficin has selective effects on some proteins so it can be a good candi-date for digestion of casein and making related drugs.

Background: Primary culture takes place following the cell isolation from tissues. Isolation and culture of melanocytes based on their roll in the protection of body against hazardous sun rays, production of skin, cornea and hair color is really important. This study was done to set isolation, culture and proliferation of melanocytes from children foreskin and adult eyelashes, and also comparison of two types of melanocyte culture medium. Methods: Human foreskin and eyelash samples were used for melanocyte isolation and culture. After isolation of epidermis from dermis, epidermis cell suspensions were prepared by enzymatic digestion. The isolated cells were cultured in two melanocyte selective culture media. Immunocytochemistary and reverse transcription-polymerase chain reaction (RT-PCR) assays were used for confirmation of isolated and cultured melanocytes. Results: Our results indicated that isolated melanocyte cultured in the selective medium without phorbol esters is better than the melanocytes cultured in selective medium cont-aining phorbol esters not only morphologically but also physiologically and from the aspect of cell adhesion. In addition, the results showed that isolated melanocyte from adult eyelashes are more dendritic than melanocytes isolated from children foreskin. Conversely, our results indicated that the number of cell passages in melanocyte isolat-ed from foreskin is more than melanocytes isolated from adult eyelashes. Conclusion: Melanocytes cultured in selective medium containing convenient growth factors in absence of phorbol esters show more native physiological and adhesive properties. In addition, melanocyte isolated from younger tissues such as foreskin have better proliferative and sub-culturing properties so we suggest isolation and culture of younger tissues.

Objective: Garlic (Allium sativum) has anti-inflammatory, anti-mutagenesis, and immunomodulatory properties that modulate anti-tumor immunity and inhibit tumor growth. In this study we have examined the effect of a protein fraction isolated from fresh garlic on anti-tumor response and intra-tumor lymphocyte infiltration.Materials and Methods: In this experimental study a protein fraction was purified from fresh garlic bulbs using ultra-filtration, followed by chromatofocusing, and SDS-PAGE analysis. Anti-tumor activity was assessed by intra-tumor injection of the protein fraction and garlic extract, itself, into groups of 5 mice each. The percentage of peripheral blood and intra-tumor CD4+ and CD8+ cells were assessed by flow cytometry. Unpaired student’s t test using the SPSS program was applied for all statistical analyses.Results: Garlic extract included different type of proteins with different molecular weight. One of protein’s fraction was immunomodeulator and was composed of three single polypeptides, with molecular masses of ~10-13 kDa and different isoelectric points (pI). These molecules augmented the delayed type hypersensitivity (DTH) response compared to the control group. Intra-tumor injection of the fraction provoked a significant increase in the CD8+ subpopulation of T-lymphocytes, as well as a decrease in tumor size. The fraction increased peripheral blood CD8+ T-lymphocytes in treated animals.Conclusion: The data confirms that protein fractions purified from fresh garlic bulbs augment CD8+ T-cell infiltration into the tumor site, inhibiting tumor growth more efficiently than garlic extract. These fi ndings provide a basis for further investigations on the purified polypeptide as a useful candidate for immunomodulation and tumor treatment.

The goal of this article is to help develop, disseminate, and evaluate resources that can be used to practice and teach EBM for urology residents and continuing education of urologists to reduce the gap between research and clinical practice. Urology departments should build capacity for residents to shape the future of quality and safety in healthcare through translating evidence into practice. Cutting edge approaches require knowing how to teach Evidence-based urology, to make Bio-statistics easy to understanding and how to lead improvement at every level. The authors shared their experience about ′what works′ in a surgical department to building an Evidence-based environment and high quality of cares.

The quality of the evidence is a keystone in the understanding of Evidence Based Medicine. Randomized controlled trials (RCTs) rank first among the research designs providing clinical evidence. Knowing about the design of clinical trials is not only the cornerstone of clinical research, but also is a requirement for any clinician who wants to learn about new findings of clinical research in his/her field. Many clinicians have good understanding as well as some misunderstandings about the design of clinical trials. This article is going to provide some crucial comments to be considered in conducting RCTs in order to help in production of better evidence for future of urology research through RCTs

Angiogenesis, the formation of new blood vessels, is a key process in cancer developmentand metastasis. In this study, the anti-angiogenic and anti-proliferative potentials of Ficuscarica latex extract have been investigated using human umbilical vein endothelial cells(HUVECs).Different doses of latex extract were prepared and added to a three-dimensional culture ofHUVEC in a collagen matrix. After 3-5 days of treatment, the anti-angiogenic effects of theextracts were monitored microscopically. For the anti-proliferation assay, different dosesof the extracts were examined on HUVECs.The results clearly indicated that latex extract could inhibit proliferation and capillary tubeformation of HUVECs in a dose-dependent manner at the range of 100-400 μg/ml. In addition,the extract was not cytotoxic up to 450 μg/ml as assessed by trypan blue and lactatedehydrogenase (LDH) cytotoxicity assays.It is concluded that latex extracts of F. carica contain strong anti-angiogenic andanti-proliferative activities. Our data indicates that latex extract could be a candidateas a potential agent for the prevention of angiogenesis in cancer and other chronicdisorders.

Objective: The significant factor contributing to the distant invasion of cancer cells is theability of tumors to produce large numbers of new blood vessels, known as angiogenesis.Many natural products inhibit angiogenesis. Herein, ethanol extract of Salvia officinalis(SO) has been analyzed for its anti-angiogenic, anti-proliferation and anti-migration activities.Materials and Methods: The anti-angiogenic effect of the SO extract was evaluated onchicken chorioallantoic membrane (CAM) neovascularisation model, microscopically. Theinhibitory effect of the extract on human umbilical vein endothelial cells (HUVECs) migrationwas tested on the wound-healing model with an inverted microscope. In addition,SO extract was screened for its possible anti-proliferative effects by separately countingHUVECs, Wehi and K562 cells with cell counter against their control wells.Results: So extract exhibited a significant inhibitory activity in CAM assay in a dose dependentmanner. CAM angiogenesis was gradually prevented to from at 100 μg/ml of SOextract, but completely inhibited to form at 200 μg/ml. After human umbilical vein endothelialcells (HUVECs) were suppressed by dose-dependent SO extract, their migrations weredetected by wound healing model, yet they were unable to show a dose response effecton proliferation of the different cells (50-200 μg/ml). As observing in this study, SOextractcould inhibit proliferation of the different cells at the concentrations above 200 μg/ml withouttoxic effect on the cells in doses ranged from 0-500 μg/ml.Conclusion: These findings indicated that SO extract might be a promising candidate foranti-angiogenic treatment.

Angiogenesis, the development of new blood vessels from existing vasculature, is essentialin physiological processes such as growth and development, wound healing andreproduction. It is also involved in pathological conditions such as tumor growth, metastases,and certain chronic diseases. Angiogenesis is dependent on a delicate equilibriumbetween endogenous angiogenic and antiangiogenic factors. However, under pathologicalconditions, this tight regulation becomes lost which can result in the formation of thedifferent diseases, including corneal neovascularization, endometriosis, obesity, atherosclerosis,diabetic retinopathy, psoriasis and cancer. In general, the process of angiogenesisis a multi-factorial and highly structured sequence of cellular events comprising migration,proliferation and differentiation of endothelial cells and finally vascular formation,maturation and remodelling. Due to the critical role of angiogenesis in physiological andpathological conditions, scientists have designed various experimental models to assayangiogenesis in vitro, ex vivo and in vivo. Currently, these experimental models are usedby many researchers for several applications such as discovering angiogenic and antiangiogenicagents, and thereby for future therapeutic applications.

Abstract Plant nonspecific lipid transfer proteins (nsLTPs) are divided into nsLTP1 and nsLTP2. The existence of an internal hydrophobic cavity, is a typical characteristic of nsLTPs that serves as the binding site for lipid substrates. In this communication a simple, rapid and low-cost alternative method was developed for purification of nsLTP2 from rice paddy. After extracting, final supernatant was loaded on CM-Sepharose column, which had previously equilibrated with 0.05 M Tris-HCl buffer, pH 8. Bounded proteins were separated using a linear gradient of 0-0.5 M NaCl. Solution of separated proteins was dialyzed and applied on a Phenyl-Sepharose column which previously equilibrated with Tris-HCl 0.05 M, ammonium sulfate 1.5 M, EDTA 0.005 M and NaHSO3 0.3%, pH 8.4. Tris-Tricin SDS-PAGE of separated proteins, obtained from ionexchange column, showed multiple bands in the range of 2-20 kDa. Further purification using hydrophobic column resulted in single band of nsLTP2 at about 7 kDa, reflecting a purified sample in the gel.

Objective: Angiogenesis is a key process in the promotion of cancer and its metastasis.Many natural health products inhibit angiogenesis. Because of the lack of molecular studieson anti-tumor and anti-angiogenic effects of shallot (Allium ascalonicum); except a few clinicalstudies on other shallot properties, such as the anti-proliferative effect of shallot chloroformicextract on two tumor cell lines, the present study focuses on the anti-angiogenic effect ofaqueous shallot extract using an aorta ring model.Materials and Methods: Aortic rings were obtained by cross-sectioning, at 1-2 mm intervals,the thoracic aorta of 4-8 weeks old Wistar male rats and cultured them in a thin drop oftype I collagen gel. After 3 days of culturing and first sprouting, the extract of A. ascalonicum(from 25 to 800 μg/ml) was added to cultures. The results of anti-angiogenic activity wereinvestigated by microscope. The cytotoxicity of extract at different doses on HUVECs wasmeasured by trypan blue assay.Results: The results showed that the shallot extract has suitable anti-angiogenic effect in arange of 50 to 800 μg/ml, but in 25 μg/ml, the extract has no considerable effect. In addition,a tangible cytotoxic effect on endothelial cells at the above mentioned doses was observed.Conclusion: Our study showed that aqueous extract of A. ascalonicum bulbs has noticeableanti-angiogenic activity without toxic effect on the cells in doses that ranged from 50-800 μg/ml. Therefore, A. ascalonicum can be a potential candidate for further investigations used inangiogenesis-related pathologic conditions.

Introduction & Objective: Egg white contains four high-quantity proteins which have numerous applications. In this research, a simple and efficient method for the purification of those proteins was designed and performed based on ion exchange chromatography. Materials & Methods: In this experimental study egg white was initially separated from insoluble substances by acidic pH. The resulting extract was isolated after two steps of ion exchange chromatography using CM-Sepharose and DEAE-Sepharose columns, respectively. Purification degree and yield of each fraction were analyzed by electrophoresis densitometry. Results: The results showed that purification degrees of ovalbumin, ovotransferrin, ovomucoid and lysozyme were 97, 98, 85 and 99 percent and their yields were 98, 98 95 and 99 percent, respectively. Conclusion: High yields, reproducibility and feasibility on low or high scales are considered as the strengths of this method.

(Received 22 Aug, 2008; Accepted 24 Sep, 2008) Abstract Background and purpose: In spite of the increasing progress in tumor treatment by current methods like surgery, chemotherapy and etc, medical sciences are unable to treat tumors. In this respect, immunology has opened a new window for tumor treatment; nowadays tumor immunotherapy is an accepted strategy for treatment of some tumors at least in some animal models. The goal of this study is the evaluation of immunotherapy using gp96- tumor peptide complex and its combination with naloxon as an opioid receptor antagonist to achieve of cellular immunity against tumorsMaterials and methods: In this study firstly, gp96 – tumor peptide complexes were purified from WEHI164 cells line using srivastava method. In the next stage, the mice, made tumoric befor by the injection of tumor cells, then were divided in to four groups. Control group were injected by PBS, test group1 were injected by naloxon, test group2 were injected by gp96 – tumor peptide complex and test group3 were injected by combination of naloxon and gp96 – tumor peptide complex. To evaluation the efficacy of vaccination, after several days, tumor volume was recorded; then the mice were killed and the spleanic cells were extracted in sterile condition. MTT test was done for cells proliferation study. Supernatant of cultured cells were collected and assayed by ELISA kits for measuring IL-4 and IFN-γ.Results: Result of protein purification had showen , purified gp96 Isoform has Molecular Weight of 66 kilo dalton.Results of tumor volume had shown that, there is no significant difference between test and control groups. Results of MTT test had shown that, there is no significant difference between test and control groups. IL-4 assay study had showed that, there is no significant difference between test group1, group2 and control group but test group3 has significantly decreased in IL-4 amount when compared with control group. Results of IFN-γ assay showed that, there is no significant difference between test group1 and control group, but test group2 and group3 has significantly increased in IFN-γ amount when compared with control group.Conclusion: It can be concluded from this study is that, prophylactic immunotherapy of tumor by combination of gp96-tumor peptide complex and naloxon, can increase IFN-γ, and, probably in a higher dosage, it may stimulate immune system more to become more potent to even decrease tumor volume. J Mazand Univ Med Sci 2008; 18(66): 1-9 (Persian)

Introduction & Objective: Isolation of cells from different tissues rely on proteolytic enzymes mainly collagenases that selectively digest collagen fibers of extra-cellular matrix. It is important to find new and proper collagenases from plant sources. In the present research actinidin, a cysteine protease abundant in Kiwifruit, was used to isolate and culture of rat hepatocytes. Materials & Methods: Different concentrations of actinidin was used to isolate rat hepatocytes by one or two-step perfusion method. The viability of the separated cells was examined by the trypan blue test. The isolated rat hepatocytes were cultured on collagen coated plates in William´s E medium. The morphology of hepatocytes was examined microscopically after staining with the Papanicolaou method. Results: Actinidin in the concentration of 0.4 mg/ml in two-step perfusion method properly isolated hepatocytes from rat liver. The viability of isolated hepatocytes that successfully cultured in collagen coated plates was 90-95 percent. Conclusion: These results showed that actinidin is a proper protease for isolation of hepatocytes. In addition, purification of this enzyme is simpler than the collagenases.

Western blotting or immunoblotting commonly use for study of reaction between antigens and antibodies. Denaturation of many proteins in immunoblotting can affect greatly the reactivity of antibodies and outcome of the procedure. In this study proteins of Brucella abortus (S19) was extracted by a mild method and reaction of the extracted proteins with serum of infected human and goat and immunized rabbit compared by affinity chromatography and immunoblotting. Gamma globulin (mostly IgG) fraction of the sera was precipitated by half saturation of ammonium sulfate and linked to activated sepharose 4B. The extracted proteins were loaded on the affinity column. Attached proteins was eluted by low pH and analyzed by SDS-PAGE. Reaction of the total extract and eluted fractions with IgG fraction of sera was evaluated by Western blotting. Upon the results of affinity chromatography and immunoblotting , Brucella proteins can be classified in four groups: 1- The proteins that adsorbed to the affinity column and react with IgG in westernblotting. 2- Proteins that react with IgG in native state but no in denatured state. 3- Proteins that do not react with IgG in native state but react in denatured state. 4- Proteins that do not react with IgG in native and denatured state.

Natural products are excellent resources for finding lead structures for the development of chemotherapeutic agents. Coumarins are a class of natural compounds found in a variety of plants. In this study, we evaluated the cytotoxic potential of coumarins isolated from Prangos ferulacea (L.) Lindl. in PC3, SKNMC, and H1299 (p53 null) human carcinoma cell lines. Osthole proved to be an outstanding potent cytotoxic agent especially against PC3 cells. Isoimperatorin exhibited moderate inhibitory effect against SKNMC and PC3 cell lines. Oxypeucedanin and braylin did not display any cytotoxic activity. In the next set of experiments, the apoptotic potentials of osthole and isoimperatorin were investigated. Induction of apoptosis by isoimperatorin was accompanied by an increase in activation of caspase-3, -8, and -9 in SKNMC cells and caspase-3 and -9 in PC3 cells. Moreover, isoimperatorin induced apoptosis by upregulating Bax and Smac/DIABLO genes in PC3 and SKNMC cells. Osthole induced apoptosis by downregulating antiapoptotic Bcl-2 in only PC3 cells and upregulating the proapoptotic genes Bax and Smac/DIABLO in PC3, SKNMC, and H1299 cells. The effects of osthole on H1299 cells are important because the loss of p53 has been associated with poor clinical prognosis in cancer treatment.