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311.
Škalamera D Ranall MV Wilson BM Leo P Purdon AS Hyde C Nourbakhsh E Grimmond SM Barry SC Gabrielli B Gonda TJ 《PloS one》2011,6(5):e20057
In response to the growing need for functional analysis of the human genome, we have developed a platform for high-throughput functional screening of genes overexpressed from lentiviral vectors. Protein-coding human open reading frames (ORFs) from the Mammalian Gene Collection were transferred into lentiviral expression vector using the highly efficient Gateway recombination cloning. Target ORFs were inserted into the vector downstream of a constitutive promoter and upstream of an IRES controlled GFP reporter, so that their transfection, transduction and expression could be monitored by fluorescence. The expression plasmids and viral packaging plasmids were combined and transfected into 293T cells to produce virus, which was then used to transduce the screening cell line. We have optimised the transfection and transduction procedures so that they can be performed using robotic liquid handling systems in arrayed 96-well microplate, one-gene-per-well format, without the need to concentrate the viral supernatant. Since lentiviruses can infect both dividing and non-dividing cells, this system can be used to overexpress human ORFs in a broad spectrum of experimental contexts. We tested the platform in a 1990 gene pilot screen for genes that can increase proliferation of the non-tumorigenic mammary epithelial cell line MCF-10A after removal of growth factors. Transduced cells were labelled with the nucleoside analogue 5-ethynyl-2'-deoxyuridine (EdU) to detect cells progressing through S phase. Hits were identified using high-content imaging and statistical analysis and confirmed with vectors using two different promoters (CMV and EF1α). The screen demonstrates the reliability, versatility and utility of our screening platform, and identifies novel cell cycle/proliferative activities for a number of genes. 相似文献
312.
Saadi Aram Dalir-Naghadeh Bahram Asri-Rezaei Siamak Anassori Ehsan 《Biological trace element research》2020,194(2):401-409
Biological Trace Element Research - Unlike in human medicine, information on the platelet selenium (Se) concentration and glutathione peroxidase (GPx) activity, as potential biomarkers of Se... 相似文献
313.
Zheng Ting Zhang Kekun Sadeghnezhad Ehsan Jiu Songtao Zhu Xudong Dong Tianyu Liu Zhongjie Guan Le Jia Haifeng Fang Jinggui 《Molecular biology reports》2020,47(10):7349-7363
Molecular Biology Reports - Chitinases (Chi), an important resistance-related protein, act against fungal pathogens by catalyzing the fungal cell wall, whereas are involved in different biological... 相似文献
314.
Behnam Honarvar Neda Odoomi Mohsen Moghadami Parvin Afsar Kazerooni Alireza Hassanabadi Parvin Zare Dolatabadi Ehsan Farzanfar Kamran Bagheri Lankarani 《PloS one》2013,8(12)
Objective
Iran has the highest rate of opiate use worldwide. However, most opiate users are not screened for hepatitis virus infections. This study aimed to provide accurate, detailed data on the size of the opiate user population at risk of developing these infections.Method
This seroprevalence study was conducted in the city of Shiraz, southern Iran. All participants were screened for HBV, HCV and HIV infection. The data were analyzed with SPSS.Result
Among 569 participants, 233 (40.9%) were injection drug users (IDU), 369 (64.8%) were heterosexual, 84 (14.7%) were bisexual and 15 (2.6%) were homosexual. One hundred nine (19.1%) were HCV antibody-positive, 18 (3.1%) were HBS antigen-positive, 72 (12.6%) were HBc antibody-positive and 23 (4%) were HIV-positive. Among IDU compared to non-IDU, positivity rates for HBS antigen (5.5 vs 1.4%), HBc antibody (22.7 vs 5.6%), HCV antibody (40.3 vs 4.4%) and HIV (7.7 vs 1.4%) were higher (P < 0.05). Most patients with HBV (80.7%) and HCV infection (83.4%) were HIV-negative. In the cumulative analysis, only history of imprisonment was a statistically significant determinant of infection by HCV or HBV in opiate users.Conclusion
The current policy of screening only HIV-positive drug users for HBV and HCV in Iran misses most cases of HBV and HCV infection. We therefore recommend urgent revision of the nationwide protocol by the Ministry of Health in Iran to implement routine screening of all opiate users and especially IDU for these viruses, regardless of their HIV status. 相似文献315.
Gerardo A. Morfini Daryl A. Bosco Hannah Brown Rodolfo Gatto Agnieszka Kaminska Yuyu Song Linda Molla Lisa Baker M. Natalia Marangoni Sarah Berth Ehsan Tavassoli Carolina Bagnato Ashutosh Tiwari Lawrence J. Hayward Gustavo F. Pigino D. Martin Watterson Chun-Fang Huang Gary Banker Robert H. Brown Jr Scott T. Brady 《PloS one》2013,8(6)
Dying-back degeneration of motor neuron axons represents an established feature of familial amyotrophic lateral sclerosis (FALS) associated with superoxide dismutase 1 (SOD1) mutations, but axon-autonomous effects of pathogenic SOD1 remained undefined. Characteristics of motor neurons affected in FALS include abnormal kinase activation, aberrant neurofilament phosphorylation, and fast axonal transport (FAT) deficits, but functional relationships among these pathogenic events were unclear. Experiments in isolated squid axoplasm reveal that FALS-related SOD1 mutant polypeptides inhibit FAT through a mechanism involving a p38 mitogen activated protein kinase pathway. Mutant SOD1 activated neuronal p38 in mouse spinal cord, neuroblastoma cells and squid axoplasm. Active p38 MAP kinase phosphorylated kinesin-1, and this phosphorylation event inhibited kinesin-1. Finally, vesicle motility assays revealed previously unrecognized, isoform-specific effects of p38 on FAT. Axon-autonomous activation of the p38 pathway represents a novel gain of toxic function for FALS-linked SOD1 proteins consistent with the dying-back pattern of neurodegeneration characteristic of ALS. 相似文献
316.
Alvandi Amirhooshang Farajzadeh Ahmad Ghaforian Borojerdnia Mehri Jelodar Abbass Aryan Ehsan Gholipour Abolfazl Masjedizadeh Abdolrahim Makvandi Manoochehr 《World journal of microbiology & biotechnology》2011,27(4):969-974
UreB is one of the urease subunits of Helicobacter pylori and can be used as an excellent antigen candidate for H. pylori vaccination. Easy access to highly purified UreB protein, facilitate advances in therapeutic or preventive strategies. To
achieve a simplified purification procedure, the present report represents a novel method of producing recombinant urease
subunit B extracellularly. ureB gene from 26,695 standard strain was amplified by PCR and cloned into pET-26b(+) expression vector. UreB was expressed as
a soluble, N-terminal pelB and C-terminal hexahistidine-tagged fusion protein (UreB-6His) and secreted into the periplasmic
space of Escherichia coli. Expression of the recombinant UreB in E. coli BL21 (DE3) was induced by isopropylthio-β-d-galactoside (IPTG). Expression of UreB was confirmed by sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE)
and western blot analysis using anti-His monoclonal antibody. UreB-6His protein was extracted from the periplasm by osmotic
shock treatment and was purified in one step by Nickel affinity chromatography. In conclusion, the present protocol is easier
to perform; more time effective and low cost than earlier methods. 相似文献
317.
Ehsan Ebrahimi Mohammad Saeed Heydarnejad Masoud Sattari Ali Bani Parnaz Kashefi 《Acta ethologica》2013,16(2):77-82
Although there exists a large body of knowledge on the ecology, physiology, and nutrition of great sturgeon, time–place learning has thus far not been examined in this species and other sturgeons. The aim of the present study is to evaluate whether individual or grouped juvenile great sturgeons are able to associate time and place of food delivery when examined under light and dark conditions. During a 30-day period, food was made available to the sturgeons twice a day or twice a night, in the first time in one side of the tank and in the second time in the opposite side of the tank. Food was withheld on test days to assess time–place learning by the fish. The results of the present study indicate that, at both the group and individual levels, great sturgeons are not able to clearly demonstrate time–place learning under the conditions examined. 相似文献
318.
Negar Firouzabadi Nader Tajik Ehsan Bahramali Hooman Bakhshandeh Mohsen Maadani Massoumeh Shafiei 《Molecular biology reports》2013,40(8):4959-4965
Etiological factors for coronary artery disease (CAD) involve a wide range of gene and environmental interactions. One of the systems being implicated in the pathophysiology of CAD is the renin-angiotensin system (RAS). However, the genetic polymorphisms of this system have not been widely studied in Iranian patients diagnosed with CAD. The aim of this study was to assess the relationship between six gene polymorphisms of RAS components and CAD in a sample of Iranian population. A total of 374 participants were enrolled in a case/control study. The presence of CAD was determined by coronary angiography. Genotyping of six RAS gene polymorphisms was performed using a modified PCR–RFLP method. Our results revealed, for the first time, a significant independent association of angiotensin-converting enzyme (ACE) A-240T polymorphism and incidence of CAD among Iranian women (P = 0.005, OR = 20.4, 95 % CI = 2.49–41.2). There has also been a significant difference in genotype distribution of ACE A-240T (P = 0.008) and angiotensin II receptor type 2 C3123A polymorphism (P = 0.032) in Iranian female participants. In conclusion, TT genotype of ACE A-240T seems to be a genetic risk factor for CAD in Iranian women. 相似文献
319.
Abdolreza Ardeshirylajimi Peyman Dinarvand Ehsan Seyedjafari Lida Langroudi Fatemeh Jamshidi Adegani Masoud Soleimani 《Cell and tissue research》2013,354(3):849-860
Tissue engineering with a combination of stem cells and nanofibrous scaffolds has attracted interest with regard to bone regeneration applications. In the present study, human induced pluripotent stem cells (iPSCs) were cultured on polymeric nanofibrous polyethersulfone (PES) with and without plasma treatment. The capacity of PES and plasma-treated PES (Plasma-PES) scaffolds to support the proliferation and osteogenic differentiation of iPSCs was investigated by MTT assay and for common osteogenic markers such as alkaline phosphatase activity, calcium mineral deposition and bone-related genes. Plasma-PES scaffolds with or without iPSCs were subsequently used to evaluate bone regeneration of critical-size defects in the rat by digital mammography, multislice spiral-computed tomography imaging and histological analysis. The results of in vitro analysis showed that plasma treatment significantly enhanced iPSC proliferation and osteogenesis. After 8 weeks of iPSC-loaded Plasma-PES implantation, no mortality or complication was observed in animals or at the site of surgery. Imaging analysis revealed more extensive bone reconstruction in rats receiving nanofibers compared with untreated control groups. Moreover, Plasma-PES seeded with iPSCs induced the highest regeneration of bone defects among all groups. These findings were confirmed by histological staining. Affective osseointegration was observed in implanted scaffolds. Thus, plasma-treated nanofibrous scaffolds are suitable tissue-engineered matrices for supporting the proliferation and osteogenic differentiation of iPSCs and might also be appropriate for the reconstruction of bone defects. 相似文献
320.