[Cloning, gene expression analysis, and phylogenic relationship of dbat gene from Iranian endemic yew [Taxus baccata L.]]

Taxol is one of the most important anticancer agents that its supply mainly depends on biological processes. 10 - deacetyl baccatin III- 0- acetyl transferase [dbat] gene is one of the key genes in Taxol biosynthetic pathway and it is expected that overexpression of this gene would result in an increased production of taxol in biological systems. Investigation of dbat gene characteristics from Iranian endemic yew, Gene expression analysis and Construction of overexpression vector for this gene. First, sequence of the gene was cloned and then, sequence characteristics identified with Bioinformatics analysis. Gene expression analysis was performed under methyl jasmonate elicitation in several time-courses. Finally in order to construct overexpression vector, the sequence was cloned under the control of the CaMV35S promoter in to pCAMBIAI304. Comparison of sequence similarities indicated that this sequence is most similar to Taxus x hunnewelliana. There was one unique amino acid substitution and investigating substituted amino acid revealed that, this replacement can not change overall characteristics of the enzyme and thus it would result in a functional enzyme. Analysis of gene expression showed that in response to methyl jasmonate, gene expression levels increased to maximum rate up to 24 times at 12 hours after elicitation. Finally gene insertion in suitable site of overexpression vector, confirmed by digest with restriction enzymes. Elicitation of shoot cuttings of Taxus would be a suitable alternative for Taxiis cell cultures in gene expression analysis. Simplicity, feasibility, and deletion of time-consuming processes of cell culture are its main advantages

Association between MDR1 C3435T gene polymorphism and acute lymphoblastic leukemia [ALL] in iranian population

P-glycoprotein [P-gp], an ATP-dependent efflux pump, is a membrane protein encoded by MDR1 gene. P-gp has an important role in protection of the cell against xenobiotics and toxic compounds. Recently, a silent C3435T polymorphism in exon 26 of MDR1 has been reported to be associated with a decreased expression of P-gp in TT genotypes carriers compared with CC genotypes carriers. To evaluate the distribution of allelic variants of C3435T MDR1 in a group of healthy population in Iran and find the association between MDR1 C3435T polymorphism and the incidence of ALL, 126 patients with ALL and 139 healthy controls were included in our study and their MDR1 polymorphisms were detected by PCRRFLP assay. 71.9% of the healthy people had 3435TC genotype, 15.8% had 3435TT genotype and 12.2% had 3435CC genotype. Also, the frequency of T allele was 51.8% and C allele 48.2%. The mutant homozygous TT and TC genotypes were found to be associated with the incidence of ALL [OR=1.96 for TT genotype and OR=0.53 for TC genotype]. MDR1 C3435T polymorphism may contribute to the incidence of ALL. TT genotypes carriers are more at risk of developing ALL than other genotypes carriers

In vitro protective effects of Satureja hortensis L. essential oil and ethanolic extract on lymphocytes DNA

DNA damage and oxidative stress are widely recognized as major factors in many degenerative diseases and aging. The protective properties of Satureja hortensis L. on the rat lymphocytes DNA lesions were tested. Lymphocytes were isolated from blood samples taken from healthy rats. DNA breaks and resistance to H[2]O[2]-induced damage were measured with the comet assay. Rat lymphocytes were incubated in S. hortensis ethanolic extract [SHE] [0.05, 0.1, 0.5, 1 and 2.5 mg/ml], essential oil [SHEO] [0.05, 0.1, 0.5, 1 and 2.5 micro l/ml], H[2]O[2] [50, 100 and 200 micro M], a combination of H[2]O[2] [200 micro M] with either SHE [1, 2.5 mg/ml] or SHEO [1, 2.5 micro l/ml] at 4°C for 30 minutes. The extent of DNA migration was measured using a single-cell microgel electrophoresis technique under alkaline conditions. Treatment of rat lymphocytes with SHE or SHEO resulted in significant reduction of H[2]O[2]-induced DNA damage compared to controls. SHE exhibited a significant [p<0.01] inhibitory effect on oxidative DNA damage at 2.5 mg/ml. SHEO [1 and 2.5 micro l/ml] also showed significant inhibitory effects [p <0.01] on H[2]O[2] induced chromosomal damage. Both the ethanolic extract and the essential oil of the plant were able to reverse the oxidative damage on rat lymphocytes induced by hydrogen peroxide

[ in vitro evaluation of antimicrobial activity of NaCIO and chlorhexidine as intracanal irrigants on streptococcus faecalis]

Today most of endodontic treatments are successful and just a low percentage of them may lead to failure. This failure may be due to remaining necrotic and infected agents in root canal system which is because of the complex anatomy of root canal and the rooms not available during mechanical and chemical cleaning. So use of chemical solutions for bacterial disinfection and necrotic agent removal is recommended. The aim of this study was to compare NaCIO Chlorhexidine and normal saline solutions in eradication of S. faecalis as a resistant bacteria in routine endodontic treatments. In this experimental and In vitro study 50 single canal human extracted teeth were selected we made conventional access cavities on all of them. Then, the teeth were divided into five equal groups randomly. Next, they were sterilized in autoclave. After that, we injected S. faecalis in root canals and put them in 37°C incubator for 36 hours. After bacterial growth a culture was prepared from each tooth to determine bacterial growth rate. Finally endodontic treatment was done on the teeth. Duration, method of filling and type of files were the same in groups but the type of chemical solution was different, [group 1: Saline, group 2: Chlorhexidine, group 3: NaCIO 2.5%, group 4: NaCIO 1%, group 5: NaCIO 5.25%]. Another culture was prepared after treatment. At last the number of colony forming units before and after treatment was compared with each other. The data were analyzed by one-way ANOVA. The results showed that there was a significant difference among NaCIO [2.5%, 5.25] and other solutions in decreasing the number of bacteria in root canals after instrumentation. Normal saline, NaCIO 1% and Chlorhexidine solutions developed similar antimicrobial activities. Considering the results of this study, when we have limitations using NaCIO, we Can use Normal Saline as a safe material. In necrotic teeth, use of NaCIO 2.5% for better removal of organisms is recommended

[Anti-tumor evaluation of adenosine and cyclohexyladenosine [A1 receptor selective agonist] using potato disc assays]

In this study, adenosine and cyclohexyladenosine [CHA] were tested for possible anti-tumor activity using potato disc assays. MIC of adenosine and CHA were determined using microplate method. In potato disc assay, discs were cut of potato with specific diameter and transferred on 1.5% agar under a laminar air cabinet. 50 ?l of a mixture containing suspension of Agrobacterium tumefaciens and the solution of agents were inoculated on potato discs in plates. The plates were incubated in 25 °C for 21 days until the tumors were counted. Adenosine and CHA did not show any anti-bacterial activity. The IC50 values of adenosine and CHA against tumor were 93.78 and 19.56 microM, respectively. DPCPX [A1 selective adenosine receptor antagonist] inhibited the anti-tumor activity of adenosine and CHA, while itself did not show any effect against potato disc tumors. This study indicates that adenosine and CHA have anti-tumor activity, and this effect is mediated via A1 receptor exists in potato cell or A. tumefaciens

Evaluation of Leishmanicidal effect of Euphorbia myrisinites extract by in vitro antileishmanial assay using promastigote of Leishmania.major

In this study, the antileishmanial effect of different extracts of Euphorbia myrsinites aerial part was evaluated on promastigotes of Leishmania major in vitro. Dried and ground aerial parts of the plant were extracted using either maceration in 80% ethanol or soxhlet in methanol. Then 5 different concentrations of each extract, one positive control, one negative control, and one solvent control were prepared and placed in a 24-well plate containing 40, 000 parasites/well. The extract concentrations were 0.06, 0.12, 0.25, 0.5 and 1 mg/ml. Amphotricin B [0.5 mg/ml] was used as positive control while negative control contained only culture medium. The plate was incubated at 25°C for six days, and the amount of parasites in each well was determined on days 2, 4, and 6 of experiment microscopically using Neubar chamber. It was observed that amphotricin B and both macerated and soxhlet extracts at concentration of 1 mg/ml killed all parasites. Lower doses exhibited a dose-dependent antileishmanial activity. The ECso for macerated and soxhlet extracts in DMSO was between 0.5 and 0.25 mg/ml. The control solvents had no significant effect on the L. major promastigotes. These results indicated that both macerated and soxhlet extracts of E. myrsinites have favorable leishmanicidal activity

[Anti-tumer and cytotoxic evaluation of crocus sativus L. Stigma and petal extracts using brine shrimp and potato disc assays]

A growing body of research has confirmed that saffron stigma extract and its main constituents have anticancer properties. The aim of the present study was to evaluate the possible cytotoxic and anti-tumor activities of the ethanolic extracts of Crocus sativus stigma and petal using simple brine shrimp and potato disc assays. The MIC of stigma and petal extracts were determined using microplate method. In brine shrimp assay, for each concentration, three wells containing 10 larva were tested. After 24h, the number of dead larva were counted. In potato disc assay, discs were cut of potato with specific diameter and transferred onto 1.5% agar under a laminar air cabinet. 50 micro1 of a mixture containing suspension of Agrobacterium tumefaciens and the solution of stigma and petal extracts were inoculated on potato discs in plates. The plates were incubated at 25°C for 21 days, and the tumors were counted. The MIC of stigma extract was 10 mg/ml. The Petal extract did not show any antibacterial activity in the of concentration range 1-16 mg/ml. The IC50 values of stigma and petal extracts against tumor were 5.3 and 10.8 mg/ml, respectively. The LC50 values of stigma and petal extracts against brine shrimp were 1678.7 and 1356.8 ppm, respectively. This study indicated that the ethanolic extracts of saffron stigma and petal have anti-tumor activity and the stigma extract has more potent anti-tumor efects than the petal extract but with less toxicity