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Cariappa R Heath-Monnig E Furesz TC Kamath SG Smith CH 《The Journal of membrane biology》2002,186(1):23-30
Our laboratory has recently identified and cloned three cationic amino-acid transporters of human placenta. We have now examined the plasma membrane domain localization and functional expression of one of these transporters, hCAT-1, in a polarized epithelial cell line (MDCK). To facilitate identification of expressed protein we first transferred the hCAT-1 cDNA to a vector with C-terminal green fluorescent protein (GFP). The resultant hCAT-1-CT-GFP fusion protein stimulated L-[3H] lysine uptake in Xenopus oocytes. In confluent monolayers of stably transfected cells grown on porous nitrocellulose filters, saturable uptake of L-[3H] lysine from the basolateral surface was stimulated 7-fold over that of untransfected cells. Concentration-dependence studies in Na+-free medium at pH 7.4 demonstrated a Km of approximately 68 +/- 13 microM and a Vmax of 970 +/-170 pmol/mg protein/min. Uptake from the apical plasma membrane surface was negligible in both transfected and untransfected cells. Consistent with these results, confocal microscopy of confluent monolayers of hCAT-1-CT-GFP-expressing cells revealed localization of the transporter solely on the basolateral domain of the cell. This is apparently the first report of a cultured polarized epithelial cell model for stable expression of a cationic amino-acid transporter. It has the potential to aid in the identification of targeting signals for transport protein localization. 相似文献
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Fuhai Li Xiaobo Zhou Jinmin Zhu Jinwen Ma Xudong Huang Stephen TC Wong 《BMC biotechnology》2007,7(1):66
Background
High content screening (HCS)-based image analysis is becoming an important and widely used research tool. Capitalizing this technology, ample cellular information can be extracted from the high content cellular images. In this study, an automated, reliable and quantitative cellular image analysis system developed in house has been employed to quantify the toxic responses of human H4 neuroglioma cells exposed to metal oxide nanoparticles. This system has been proved to be an essential tool in our study. 相似文献26.
TL Kieft EM Murphy DL Haldeman PS Amy BN Bjornstad EV McDonald DB Ringelberg DC White J Stair RP Griffiths TC Gsell WE Holben DR Boone 《Microbial ecology》1998,36(3):336-348
Abstract Two chronosequences of unsaturated, buried loess sediments, ranging in age from <10,000 years to >1 million years, were investigated to reconstruct patterns of microbial ecological succession that have occurred since sediment burial. The relative importance of microbial transport and survival to succession was inferred from sediment ages, porewater ages, patterns of abundance (measured by direct counts, counts of culturable cells, and total phospholipid fatty acids), activities (measured by radiotracer and enzyme assays), and community composition (measured by phospholipid fatty acid patterns and Biolog substrate usage). Core samples were collected at two sites 40 km apart in the Palouse region of eastern Washington State, near the towns of Washtucna and Winona. The Washtucna site was flooded multiple times during the Pleistocene by glacial outburst floods; the Winona site elevation is above flood stage. Sediments at the Washtucna site were collected from near surface to 14.9 m depth, where the sediment age was approximately 250 ka and the porewater age was 3700 years; sample intervals at the Winona site ranged from near surface to 38 m (sediment age: approximately 1 Ma; porewater age: 1200 years). Microbial abundance and activities declined with depth at both sites; however, even the deepest, oldest sediments showed evidence of viable microorganisms. Same-age sediments had equal quantities of microorganisms, but different community types. Differences in community makeup between the two sites can be attributed to differences in groundwater recharge and paleoflooding. Estimates of the microbial community age can be constrained by porewater and sediment ages. In the shallower sediments (<9 m at Washtucna, <12 m at Winona), the microbial communities are likely similar in age to the groundwater; thus, microbial succession has been influenced by recent transport of microorganisms from the surface. In the deeper sediments, the populations may be considerably older than the porewater ages, since microbial transport is severely restricted in unsaturated sediments. This is particularly true at the Winona site, which was never flooded. 相似文献
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Two cases of subacute sclerosing panencephalitis treated with 5-bromo-2-deoxyuridine (BUDR) are reported. Both cases had a classical presentation with motor and mental deterioration, myoclonic jerks, paretic colloidal gold curve in CSF and periodic bursts of the EEG. The diagnosis was confirmed by brain biopsy with light and electron microscopic findings. Immunological studies revealed markedly elevated serum and CSF measles antibodies on serial determinations by the virus neutralization, complement fixation and hemagglutination inhibition techniques. Both cases were treated with 100 mg./kg./day of BUDR, intravenously, for five days, and Case 2 received a second course of treatment. Only minimal side effects were experienced from the use of BUDR; clinical symptoms showed sustained improvement in Case 1 and no further deterioration in Case 2. Both patients survived for more than 16 months. More extensive controlled trials with antiviral agents for the treatment of subacute sclerosing panencephalitis appear to be justified. 相似文献
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Hong Zhao Jinmin Zhu Kemi Cui Xiaoyin Xu Megan O'Brien Kelvin K Wong Santosh Kesari Weiming Xia Stephen TC Wong 《Cancer cell international》2009,9(1):15
Background
Cancer and Alzheimer's disease (AD) are two seemingly distinct diseases and rarely occur simultaneously in patients. To explore molecular determinants differentiating pathogenic routes towards AD or cancer, we investigate the role of amyloid β protein (Aβ) on multiple tumor cell lines that are stably expressing luciferase (human glioblastoma U87; human breast adenocarcinoma MDA-MB231; and mouse melanoma B16F). 相似文献30.
Tobamoviruses, mostly isolated from solanaceous plants, may represent
ancient virus lineages that have codiverged with their hosts. Recently
completed nucleotide sequences of six nonsolanaceous tobamoviruses allowed
assessment of the codivergence hypothesis and support a third subgroup
within tobamoviruses. The genomic sequences of 12 tobamoviruses and the
partial sequences of 11 others have been analyzed. Comparisons of the
predicted protein sequences revealed three clusters of tobamoviruses,
corresponding to those infecting solanaceous species (subgroup 1), those
infecting cucurbits and legumes (subgroup 2), and those infecting
crucifers. The orchid-infecting odontoglossum ringspot tobamovirus was
associated with subgroup 1 genomes by its coat and movement protein
sequences, but with the crucifer-pathogenic tobamoviruses by the remainder
of its genome, suggesting that it is the progeny of a recombinant. For four
of five genomic regions, subgroup 1 and 3 genomes were equidistant from a
subgroup 2 genome chosen for comparison, suggesting uniform rates of
evolution. A phylogenetic tree of plant families based on the tobamoviruses
they harbor was congruent with that based on rubisco sequences but had a
different root, suggesting that codivergence was tempered by rare events of
viruses of one family colonizing another family. The proposed subgroup 3
viruses probably have an origin of virion assembly in the movement protein
gene, a large (25-codon) overlap of movement and coat protein open reading
frames, and a comparably shorter genome. Codon-position- dependent base
compositions and codon prevalences suggested that the coat protein frame of
the overlap region was ancestral. Bootstrapped parsimony analysis of the
nucleotides in the overlap region and of the sequences translated from the
-1 frame (the subgroup 3 movement protein frame) of this region produced
trees inconsistent with those deduced from other regions. The results are
consistent with a model in which a no or short overlap organization was
ancestral. Despite encoding of subgroup 2 and 3 movement protein C-termini
by nonhomologous nucleotides, weak similarities between their amino acid
sequences suggested convergent sequence evolution.
相似文献