The Open Chemical and Biomedical Methods Journal最新文献

{"title":"siRNA-Lipoplex-Mediated Bcl-2 and Bcl-xL Gene Silencing Induces Apoptosis in MCF-7 Human Breast Carcinoma Cells","authors":"Assaf Vestin, E. Khazanov, D. Avni, V. Sergeyev, Y. Barenholz, Y. Sidi, E. Yakobson","doi":"10.2174/1875038900801010028","DOIUrl":"https://doi.org/10.2174/1875038900801010028","url":null,"abstract":"Bcl-2 family genes play a central role in cell apoptosis and cell proliferation, and are implicated in the pathol- ogy of many malignancies. We explored different ways of introducing siRNA duplexes into cells, comparing \"naked\" with lipoplex- and polyplex-based formulations in order to decrease the level of the Bcl-2 and Bcl-xL proteins. Our results show that siRNA binds efficiently to all cationic liposomes used. Upon binding, siRNA reduces the zeta potential of the particles, although in most cases they remain positively charged. 70% of MCF-7 cells took up fluorescently-labeled siRNA after 24 h. All siRNA sequences caused growth inhibition of cells, with variable efficiency in a dose-dependent manner. Significant decreases in Bcl-2 and Bcl-xL proteins were caused by two siRNA sequences. Both caused signifi- cant growth inhibition in concentrations as low as 100 nM. These two siRNAs caused the greatest increase in caspase-7 activity and DNA fragmentation level. Addition of CaCl 2 as a transfection enhancer resulted in marked increase of growth inhibition and Bcl-2 gene suppression by siRNA. Our lipoplexes containing siRNA showed equal or superior efficacy in comparison with commercial siRNA transfection kits. Efficiency of cell growth inhibition per RNA molecule using siRNA was found to be twenty fold higher than by a well-established Bcl-2 antisense oligonucleotide (ODN) molecule af- ter optimization of ODN delivery to cells in culture. This study indicates the potential for efficient delivery of siRNA for treatment of various malignancies.","PeriodicalId":302199,"journal":{"name":"The Open Chemical and Biomedical Methods Journal","volume":"18 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"117145657","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A Simple Sample Preparation with HPLC-UV Method for Estimation of Amlodipine from Plasma: Application to Bioequivalence Study","authors":"S. Meyyanathan, S. Muralidharan, S.Rajan, K. Gopal, B. Suresh","doi":"10.2174/1875038900801010022","DOIUrl":"https://doi.org/10.2174/1875038900801010022","url":null,"abstract":"A simple, rapid and selective method was developed for the estimation of amlodipine from human plasma. The method involves a simple protein precipitation techniques using nifedipine as internal standard. Chromatographic separa- tion was carried out on a reverse phase C18 column using mixture of 50 mM potassium di hydrogen ortho phosphate (pH 7.5) and acetonitrile (60:40, v/v) at a flow rate of 1.0 mL/min with UV detection at 239 nm. The retention time of am- lodipine and internal standard were 4.12 and 8.31min, respectively. The method was validated and found to be linear in the range of 0.5-50.0 ng/mL. An open, randomized, two-treatment, two period, single dose crossover, bioequivalence study in 24 fasting, healthy, male, volunteers was conducted. After dosing, serial blood samples were collected for the pe- riod of 168.0 h. Various pharmacokinetic parameters including AUC0-t, AUC0-�, Cmax, Tmax, T1/2, and elimination rate constant (Kel) were determined from plasma concentration of both formulations of test (Amlodipine 5 mg tablets) and ref- erence (Amlodipine 5 mg tablets). Log transformed values were compared by analysis of variance (ANOVA) followed by classical 90% confidence interval for Cmax, AUC0-t and AUC0-� and was found to be within the range. These results indi- cated that the analytical method was linear, precise and accurate. Test and reference formulation were found to be bioe- quivalent.","PeriodicalId":302199,"journal":{"name":"The Open Chemical and Biomedical Methods Journal","volume":"3 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"122586907","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"HPLC assessment and multivariate predictability of serum retinol and α-tocopherol concentrations in adult female subjects","authors":"A. Jaworowska, G. Bazylak","doi":"10.2174/1875038900801010011","DOIUrl":"https://doi.org/10.2174/1875038900801010011","url":null,"abstract":"Antioxidant vitamins have been reported to protect against a variety of human malignances and multiple chronic degenerative diseases therefore it is important to understand factors that influence their blood levels. The pre- sent study was conducted to verify association of serum retinol and � -tocopherol levels with obesity, and to assess predic- tors of their serum concentrations in representative sample population of overweight/obese (n = 51) and normal weight (n = 26) apparently healthy adult female subjects recruited from typical urban area in Poland. Anthropometric measurements were taken from all participants who also completed a questionnaire on selected lifestyle factors. The serum concentra- tions of retinol and � -tocopherol were measured by fully validated Chromsystems diagnostic kit employing isocratic RP- HPLC with switched wavelength UV detection. Intake of energy, fat, vitamin A and E and alcohol consumption were es- timated by seven daily dietary records. Multivariate linear regression models were fitted in order to estimate the predictors of serum retinol and � -tocopherol concentration. There were no statistically significant differences in the average se- rum levels of retinol and � -tocopherol between overweight/obese and normal weight adult female subjects. Serum � - tocopherol concentrations were positively correlated with serum total cholesterol level (TChol) and body mass index (BMI), but inversely with total energy intake and past dieting behaviour. The TChol and total energy intake were identi- fied as predictors of serum retinol levels. Intakes of fat and vitamin E and A, age, serum triglyceride concentration, smok- ing, alcohol consumption and physical activity were unrelated to serum levels of retinol or � -tocopherol. The main finding of this study is that obesity is not associated with decreased serum retinol and � -tocopherol levels. In addition, these re- sults indicated that dietary intake of vitamin A and E are poor predictors for serum retinol or � -tocopherol concentra- tions. Serum levels of both these vitamins are primarily influenced by TChol, obesity, total energy intake and past dieting behaviour.","PeriodicalId":302199,"journal":{"name":"The Open Chemical and Biomedical Methods Journal","volume":"16 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"116851016","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"HPLC Analysis of Trans-Resveratrol in Human Plasma After Red Wine Consumption","authors":"L. L. Corre, A. Léger-Enreille, N. Chalabi, L. Delort, Y. Bignon, D. Bernard-Gallon","doi":"10.2174/1875038900801010007","DOIUrl":"https://doi.org/10.2174/1875038900801010007","url":null,"abstract":"Trans-resveratrol (t-RES), a phenolic compound produced by several plants and present in wine, has been re- ported to be a potential chemopreventive agent for cardiovascular, cancer and neurodegenerative pathologies. Thus, un- derstanding the plasma level in vivo of trans-resveratrol is the prerequisite to evaluate its potential health impact. Bioavailability studies mainly in animals or in humans using the pure compound at very high doses were performed. The objective of this present study was to detected trans-resveratrol in human plasma from two subjects who consumed 600 mL of red wine over 40 min. Plasma analyses were performed by HPLC and obtained results indicated the absence of t- RES in subject plasma at any time with limit of detection of 5 ng/mL. In conclusion, this study suggests that t-RES from red wine is poorly bioavailable and even an important red wine consumption does not make it possible to obtain detect- able plasma concentrations of t-RES.","PeriodicalId":302199,"journal":{"name":"The Open Chemical and Biomedical Methods Journal","volume":"23 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-06-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134242389","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Use of the Real Time Polymerase Chain Reaction Method to Establish a Tightly Regulated Ecdysone Inducible System in Mammalian Cells","authors":"R. Lai, H. Aung, T. Walsh, R. Barnard","doi":"10.2174/1875038900801010001","DOIUrl":"https://doi.org/10.2174/1875038900801010001","url":null,"abstract":"Introduction: The ecdysone inducible system potentially allows the study of conditional expression of the ex- ogenous reporter genes that may be cell lethal or alter the phenotype during the selection of transfectants. The system re- lies on two independent transfections of plasmids named pVgRXR and pIND. Disruption of the regulatory element within the plasmid during stable integration can result in silenced or high background expression of the exogenous reporter gene. Previous studies (1) have reported a transient luciferase reporter assay to screen the cell lines stably transfected with pVgRXR plasmid. However, there is no suitable method to screen the subsequent pIND transfection. In this study, we demonstrate a real time polymerase chain reaction (PCR) strategy to screen for background expression problems associ- ated with the ecdysone expression system and to simultaneously allow discrimination between the products of endoge- nously expressed and transfected genes. Method: Two screening methods were applied sequentially in order to establish a functional ecdysone expression system. Firstly, the HCT116/VgRXR#8 (a human colon cancer cell line stably transfected with pVGRXR plasmid) was estab- lished by utilising the previously reported luciferase reporter assay (1). Finally, the functional ecdysone system (HCT116/VgRXR#8/mutant p53) was established by a real time PCR screening strategy. This PCR based screening method for the exogenous reporter gene was made possible by utilizing the unique BGH polyA tail from the exogenous reporter gene. Result: Even when the same parental cell line (HCT116/VgRXR#8) was used in the subsequent transfection, background expression was still a common phenomenon. This can be monitored by efficient and sensitive real time PCR. Further- more, the primers designed in this study have high specificity for the exogenous reporter gene. Conclusion: The combined use of luciferase and real time PCR methods was necessary to enable the establishment of a tightly regulated ecdysone inducible system in mammalian cells.","PeriodicalId":302199,"journal":{"name":"The Open Chemical and Biomedical Methods Journal","volume":"19 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2008-06-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"126780903","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}