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71.
Agha-Mohammadi S O'Malley M Etemad A Wang Z Xiao X Lotze MT 《The journal of gene medicine》2004,6(7):817-828
BACKGROUND: The tetracycline-regulatable system is one of the most valuable tools for controlling gene expression. In its current form, however, the system is less than ideal for in vivo or gene therapy uses due to difficulties in set-up procedures, high basal leakiness, and unpredictable delivery and efficiency. METHODS: To address these issues, we have devised a second generation of tetracycline-regulated promoters (TREs). The second-generation TRE (SG-TRE) contains a shortened cytomegalovirus (CMV) minimal promoter together with eight tet operator sequences positioned in an optimized manner upstream of the TATA box. This construct displays far greater reduction in basal leakiness than maximal transgene expression. Conversely, maximal transgene expression is increased to a greater degree than basal leakiness by post-translational stabilization with bovine growth hormone poly A. RESULTS: In transient studies, the SG-TRE displays over 100 000-fold regulation efficiency in HeLa cells at 1:1 ratio of transactivator to reporter plasmid in the Tet-Off system. This novel promoter achieves a regulation efficiency 500- to 1000-fold higher than that of the original TRE (P(hCMV*-1)) in HeLa cells by displaying undetectable levels of basal leakiness without compromised maximal expression. In other cell lines, the SG-TRE proves to be more efficient than the original P(hCMV*-1) in a cell-dependent manner. Furthermore, the SG-TRE preserves its enhanced regulation efficiency and its reduced basal leakiness in the context of a single positive feedback regulatory vector that presents ease of delivery of the system for use in vivo. Finally, in vivo, the biological function of granulocyte-macrophage colony stimulating factor is tightly regulated in the context of SG-TRE delivered via adeno-associated viruses. 相似文献
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Mohammadi S Normand JP Turcotte R Voisine P Mathieu P Dagenais F 《Innovations (Philadelphia, Pa.)》2007,2(1):48-50
A 76-year-old man with an ascending arch and proximal descending aortic aneurysm underwent a complex aortic replacement through a sternotomy with ligation of a right aberrant subclavian artery (RASA) distal to the right vertebral artery. The second-stage procedure was performed with a stent-graft deployed within the elephant trunk. At 6- and 12-month follow-up, the RASA was opacified by the patent right vertebral artery. Under ultrasound guidance, the patient's RASA stump was occluded by coils. Management of an RASA during complex hybrid stent-graft procedures is discussed. 相似文献
75.
Janghorbani M Amini M Willett WC Mehdi Gouya M Delavari A Alikhani S Mahdavi A 《Obesity (Silver Spring, Md.)》2007,15(11):2797-2808
Objective: The goal was to estimate the prevalence of overweight, obesity, underweight, and abdominal obesity among the adult population of Iran. Research Methods and Procedures: A nationwide cross‐sectional survey was conducted from December 2004 to February 2005. The selection was conducted by stratified probability cluster sampling through household family members in Iran. Weight, height, and waist circumference (WC) of 89,404 men and women 15 to 65 years of age (mean, 39.2 years) were measured. The criteria for underweight, normal‐weight, overweight, and Class I, II, and III obesity were BMI <18.5, 18.5 to 24.9, 25 to 29.9, 30 to 34.9, 35 to 39.9, and ≥40 (kg/m2), respectively. Abdominal obesity was defined as WC ≥102 cm in men and ≥88 cm in women. Results: The age‐adjusted means for BMI and WC were 24.6 kg/m2 in men and 26.5 kg/m2 in women and 86.6 cm in men and 89.6 cm in women, respectively. The age‐adjusted prevalence of overweight or obesity (BMI ≥25) was 42.8% in men and 57.0% in women; 11.1% of men and 25.2% of women were obese (BMI ≥30), while 6.3% of men and 5.2% of women were underweight. Age, low physical activity, low educational attainment, marriage, and residence in urban areas were strongly associated with obesity. Abdominal obesity was more common among women than men (54.5% vs. 12.9%) and greater with older age. Discussion: Excess body weight appears to be common in Iran. More women than men present with overweight and abdominal obesity. Prevention and treatment strategies are urgently needed to address the health burden of obesity. 相似文献
76.
Salehi Mina Moieni Ahmad Safaie Naser Farhadi Siamak 《Journal of industrial microbiology & biotechnology》2019,46(5):613-623
Journal of Industrial Microbiology & Biotechnology - Paclitaxel is a main impressive chemotherapeutic agent with unique mode of action and broad-spectrum activity against cancers. Hazel... 相似文献
77.
lambda-Repressor oligomerization kinetics at high concentrations using fluorescence correlation spectroscopy in zero-mode waveguides
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Fluorescence correlation spectroscopy (FCS) has demonstrated its utility for measuring transport properties and kinetics at low fluorophore concentrations. In this article, we demonstrate that simple optical nanostructures, known as zero-mode waveguides, can be used to significantly reduce the FCS observation volume. This, in turn, allows FCS to be applied to solutions with significantly higher fluorophore concentrations. We derive an empirical FCS model accounting for one-dimensional diffusion in a finite tube with a simple exponential observation profile. This technique is used to measure the oligomerization of the bacteriophage lambda repressor protein at micromolar concentrations. The results agree with previous studies utilizing conventional techniques. Additionally, we demonstrate that the zero-mode waveguides can be used to assay biological activity by measuring changes in diffusion constant as a result of ligand binding. 相似文献
78.
Lai F Khojasteh-Bakht SC 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2005,814(2):225-232
In vitro metabolic stability studies are performed routinely in drug discovery to determine the rate of metabolism as well as the metabolic fate of compounds. These studies are labor intensive, involving incubation of the compound with a biological matrix, sampling at various time points, stopping the reaction, and sample preparation for analysis. All of these steps involve manual pipetting in the conventional method. An automated method for in vitro metabolism studies is reported here. The method reduces the time and manual labor required and has other advantages, such as better reproducibility and unattended operation. This method utilizes an autosampler custom configured with cooling and incubation capabilities. The autosampler is programmed to directly inject incubation samples at set time points onto an online extraction column. The extracted sample then enters an analytical column for separation and ultimately the mass spectrometer for detection. The injection has the dual function of stopping the reaction and starting the analysis on the LC-MS. This method was used for the metabolic stability study of a prodrug in plasma and liver S9 fractions of five different species. The stability data from the automated method were similar to those obtained using the conventional method. The potential for this method to increase throughput of metabolic stability studies in drug discovery is demonstrated. 相似文献
79.
Zhaohe Luo Kenneth Neil Mertens Siamak Bagheri Hilal Aydin Yoshihito Takano Kazumi Matsuoka 《欧洲藻类学杂志》2016,51(2):188-202
Species belonging to the dinophyte genus Scrippsiella are frequently reported in marine waters, but information on their distribution in brackish environments is limited. Here we describe a new species, S. plana, through incubation of non-calcified cysts from sediments collected in the South China Sea and Caspian Sea. The vegetative cells consist of a conical epitheca and a rounded hypotheca with the plate formula of Po, X, 4′, 3a, 7′′, 5C+t, 5S, 5′′′, 2′′′′. It differs from other Scrippsiella species by its flattened body in dorsoventral view and a small first anterior intercalary (1a) plate (half the size of plate 3a). Scrippsiella plana strains from the South China Sea and Caspian Sea share identical internal transcribed spacer (ITS) sequences, and show phenotypic plasticity and local adaptation in growth rate at various salinities, consistent with the environments in which they originated. In addition, two strains of S. spinifera were obtained by incubating ellipsoid cysts with calcareous spines from sediments collected along the Turkish and Hawaiian coast. They also share identical ITS sequences and differ from Duboscquodinium collinii (a parasite of tintinnids) only at two base pair positions (in the ITS2 region). Molecular phylogeny based on ITS and large subunit ribosomal DNA (LSU rDNA) sequences revealed that S. plana was nested within the Calciodinellum (CAL) clade and S. spinifera within the S. trochoidea (STR) clade. The phylogenetic position of ‘Peridinium’ wisconsinense is reported for the first time, which supports multiple transitions of the Peridiniales to freshwater. 相似文献
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Laura N. Bull Jenneke A. Juijn Mira Liao Michiel J. T. van Eijk Richard J. Sinke Nicole L. Stricker Joseph A. DeYoung Victoria E. H. Carlton Siamak Baharloo L. W. J. Klomp Daiki Abukawa David E. Barton Nathan M. Bass Billy Bourke Brendan Drumm Irena Jankowska Piero Lovisetto Shirley McQuaid Joanna Pawlowska Yusaku Tazawa Erica Villa Niels Tygstrup Ruud Berger Alexander S. Knisely Roderick H. J. Houwen N. B. Freimer 《Human genetics》1999,104(3):241-248
Loci for two inherited liver diseases, benign recurrent intrahepatic cholestasis (BRIC) and progressive familial intrahepatic
cholestasis type 1 (PFIC1), have previously been mapped to 18q21 by a search for shared haplotypes in patients in two isolated
populations. This paper describes the use of further haplotype evaluation with a larger sample of patients for both disorders,
drawn from several different populations. Our assessment places both loci in the same interval of less than 1 cM and has led
to the discovery of the PFIC1/BRIC gene, FIC1; this discovery permits retrospective examination of the general utility of haplotype evaluation and highlights possible
caveats regarding this method of genetic mapping.
Received: 21 September 1998 / Accepted: 29 December 1998 相似文献