Synthesis, and in-vitro cytotoxicity studies of a series of triazene derivatives on human cancer cell lines

Compounds containing triazene ring structure are cytotoxic agents and clinically used as antitumor alkylating agents. In this study, a series of triazene derivatives holding alkyl and aryl moieties were synthesized and proved to be potent cytotoxic agents in-vitro particularly against eight cancer cell lines [PCS, HT29, Hela, HL60, Jurkat, K562, MCF7, HepG2] and a non-cancerous cell line [HUVEC]. The cytotoxic activity was assessed using two methods, LDH assay, and trypan blue exclusion. Some of the triazene derivatives showed cytotoxic activity more than temozolomide [TMZ] as the reference drug. The synthesized triazenes showed marked cytotoxicity effects on all eight cancer cell lines. Among the compounds synthesized, l,3-bis [2-ethoxyphenyl] triazene C had unique efficacy and selectivity so that it had IC[50] between 0.560-3.33 microM on cancer cell lines and 12.61 microM on normal cell line [HUVEC]. l-[4-nitrophenyl]-3-[2-hydroxyethyl] triazene E shows weaker effect on cancer cell lines than the other compounds having 1C[50] between 3-15.54 microM

[Primary culture of human skin melanocyte and comparison of culture in the presence and absence of phorbol ester]

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. 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. Our results indicated that isolated melanocyte cultured in the selective medium without phorbol esters is better than the melanocytes cultured in selective medium containing 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 isolated from foreskin is more than melanocytes isolated from adult eyelashes. 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.

Purified protein fraction of garlic extract modulates cellular immune response against breast transplanted tumors in BALB/c mice model

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. 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. Un-paired student's t test using the SPSS program was applied for all statistical analyses. 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. Intratumor 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. 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 findings provide a basis for further investigations on the purified polypeptide as a useful candidate for immunomodulation and tumor treatment

Study of the mechanisms and causes related to angiogenesis

Angiogenesis, the process of new blood vessels formation from existing ones, is important for the normal body growth and development. Angiogenesis is dependent on a delicate equilibrium between endogenous angiogenic and anti-angiogenic factors. If this equilibrium is disturbed, different diseases such as corneal neovascularization, endometriosis, obesity, atherosclerosis, diabetic retinopathy, psoriasis, tumor growth and metastasis may ensue. In general, angiogenesis process is under the influence of several factors and consists of cellular events such as migration, proliferation and differentiation of endothelial cells and finally formation, maturation and remodeling of vessels. Hence, angiogenesis inhibition as a useful measure in conventional therapies such as chemotherapy and radiation has attracted scientists' attention studying on these issues. In recent years, the angiogenesis control has been considered as a novel idea for control and treatment of angiogenesis-dependent disorders especially tumor growth and metastasis. Because of its importance in causing these disorders, we discuss various aspects of angiogenesis, its mechanisms and causes and also the related studies in this review article

[Angiogenesis and tumor]

Angiogenesis, the process of new blood vessel formation from existing ones, plays an important role in the physiologic circumstances such as embryonic development, placenta formation, and wound healing. It is also crucial to progress of pathogenic processes of a variety of disorders, including tumor growth and metastasis. In general, angiogenesis process is a multi-factorial and highly structured sequence of cellular events comprising migration, proliferation and differentiation of endothelial cells and finally vascular formation, maturation and remodeling.Thereby, angiogenesis inhibition as a helping agent to conventional therapies such as chemotherapy and radiation has attracted the scientists' attentions studying in this field

Anti tumor activity of salvia officinalis is due to its anti-angiogenic, anti-migratory and anti-proliferative effects

The significant factor contributing to the distant invasion of cancer cells is the ability 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. The anti-angiogenic effect of the SO extract was evaluated on chicken chorioallantoic membrane [CAM] neovascularisation model, microscopically. The inhibitory effect of the extract on human umbilical vein endothelial cells [HUVECs] migration was tested on the wound-healing model with an inverted microscope. In addition, SO extract was screened for its possible anti-proliferative effects by separately counting HUVECs, Wehi and K562 cells with cell counter against their control wells, So extract exhibited a significant inhibitory activity in CAM assay in a dose dependent manner. CAM angiogenesis was gradually prevented to from at 100 micro g/ml of SO extract, but completely inhibited to form at 200 micro g/ml. After human umbilical vein endothelial cells [HUVECs] were suppressed by dose-dependent SO extract, their migrations were detected by wound healing model, yet they were unable to show a dose response effect on proliferation of the different cells [50-200 micro g/ml]. As observing in this study, SOextract could inhibit proliferation of the different cells at the concentrations above 200 micro g/ml without toxic effect on the cells in doses ranged from 0-500 micro g/ml. These findings indicated that SO extract might be a promising candidate for anti-angiogenic treatment

Anti-angiogenic activity of ficus carica latex extract on human umbilical vein endothelial cells

Angiogenesis, the formation of new blood vessels, is a key process in cancer development and metastasis. In this study, the anti-angiogenic and anti-proiiferative potentials of Ficus carica 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 of HUVEC in a collagen matrix. After 3-5 days of treatment, the anti-angiogenic effects of the extracts were monitored microscopically. For the anti-proliferation assay, different doses of the extracts were examined on HUVECs. The results clearly indicated that latex extract could inhibit proliferation and capillary tube formation of HUVECs in a dose-dependent manner at the range of 100-400 micro g/ml. In addition, the extract was not cytotoxic up to 450 micro g/ml as assessed by trypan blue and lactate dehydrogenase [LDH] cytotoxicity assays. It is concluded that latex extracts of F. carica contain strong anti-angiogenic and anti-proliferative activities. Our data indicates that latex extract could be a candidate as a potential agent for the prevention of angiogenesis in cancer and other chronic disorders

Femtosecond laser versus mechanical microkeratome in thin-flap laser in situ keratomileusis [Lasik] for correction of refractive errors an evidence-based effectiveness and cost analysis

To compare the efficacy and cost-effectiveness of Femtosecond laser versus mechanical Microkeratome corneal flap creation in correction of refractive errors. In this review, a comprehensive search of Medline, SCOPUS, Cochrane, TRIP database, supplemented by HTA and economic databases was performed. We searched for randomized controlled trials [RCTs] of Femtosecond laser which included mechanical Microkeratome in other arm. The quality of the retrieved studies was appraised by two independent reviewers and appropriate articles were finalized. A total of 1142 articles were identified, of which, 1059 were excluded after review of the titles and abstracts and 83 articles remained. Systematic reviews and RCTs were evaluated through CASP international worksheet. Eventually, 61 titles were excluded, leaving 22 articles to be reviewed. Safety: There was no individual evidence to cover all safety components about Femtosecond laser, but in summary, this modality seems a safe method for corneal flap creation. Effectiveness: No statistically significant difference was shown in visual acuity and refractive errors. The important secondary end point of this review was diffuse Lamellar keratitis in 17% of the Femtosecond group versus 5% in mechanical Microkeratome. Inflammation was low-grade and improved during the first 3 months of follow-up period with a low dose medication without corneal scarring. The two groups was comparable in all clinical outcomes including Unorrected Visual Acuity [UCVA], Best Special Corrected Visual Acuity [BSCVA], manifest refraction, wave front aberrometry, Schirmer test, and Tear Break up time [TBUT]. Cost Analysis: Results showed that marginal cost incurred due to Femtosecond technology adoption may vary from 27 to 117 _ [resulted from sensitivity analysis]. It is clear that additional cost may be a small proportion of LASIK procedure total cost. Although Femtosecond flap creation is a modern method with a good quality of corneal flap, but, there is no high-quality evidence to show superiority of Femtosecond laser in clinical outcomes. Although the efficacy and cost of the systems is almost equal, traditional method still remains as the standard approach

Holmium laser prostate enucleation [HOLEP] versus trans-urethral resection of prostate [TURP] in treatment of symptomatic prostatic enlargement; a health technology assessment

Our aim was to compare the cost effectiveness of holmium laser prostate enucleation [HOLEP] versus trans-urethral resection of prostate. We searched all available databases for any controlled trials comparing HOLEP and TURP from January 2000 to February 2009. Two independent reviewers studied and appraised the selected evidences. Then, effectiveness and cost effectiveness of HOLEP was evaluated. We identified four randomized controlled trials and one systematic review according to the inclusion criteria. Most of the studies had moderate quality of evidence with limited sample sizes. Overall success rate of HOLEP was comparable with TURP; but, some secondary outcomes such as pick flow rate twelve months after the surgery was better in HOLEP. A comparison between the original costs and those obtained from sensitivity analysis showed that the cost parameters were sensitive to the number of the patients treated. Increasing the number of the patients from 200 to 300 changed the study's results in favor of the new techniques. Since the holmium and thulium laser sets are sensitive to the number of the patients and multipurpose, they potentially can be applied for stone fragmentation. Thus, utilization of these equipments will divide the costs between two groups of the services. In economic terms, these properties lower overhead costs and justify the purchasing of these equipments

[Evaluation of cell proliferation and interleukine-4 and interferon gamma level measurement in Balb/c with experimental fibrosarcoma tumor by combination of Gp96-tumor peptide and naloxan]

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 tumors. 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 before 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- gamma. 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-gamma assay showed that, there is no significant difference between test group1 and control group, but test group2 and group3 has significantly increased in IFN- gamma amount when compared with control group. 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- gamma, and, probably in a higher dosage, it may stimulate immune system more to become more potent to even decrease tumor volume

Evaluation of cellular immune response against purified antigen 85 in patients with tuberculosis

Tuberculosis [TB] remains an important health problem throughout the world. Despite its significance in public health, mechanisms of protective immunity against Mycobacyerium Tuberculosis in humans have not yet been understood. To evaluate cell mediated immune response against purified Ag 85, PPD and Phytohemagglutinin [PHA] in patients with tuberculosis and healthy tuberculin positive and negative individuals. Thirty patients with tuberculosis and 60 healthy tuberculin skin test positive and negative volunteers were participated in this study. Cell mediated immunity was assessed by measuring [[3]H]-thymidine uptake and detection of IFN-gamma in the culture supernatant using commercial ELISA test. In the present study, we showed that IFN-gamma production and cell proliferation response to Ag 85 were significantly higher in tuberculin positive than tuberculin negative individuals [P<0.01]. Among tuberculous patients, IFN-gamma production and cell proliferative responses to Ag 85 was significantly lower in contrast to healthy tuberculin positive individuals [P<0.01]. In addition, IFN- gamma response in patients with cavitary tuberculosis was lower than patients without cavitation [P<0.05]. Based on the higher cell mediated immune responses to Ag 85 in healthy tuberculin positive volunteers compared to patients [especially with advanced disease], purified Ag 85 can be used as a sensitive marker for analysis of immune responses in tuberculosis