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311.
Anna L. Trifillis Myong Won Kahng 《In vitro cellular & developmental biology. Plant》1990,26(5):441-446
Summary We have developed an in vitro model of human papillary collecting duct cells isolated from cadaver kidneys using methods similar
to those we previously reported for the isolation of human proximal tubule cells. To date we have isolated papillary collecting
duct cells from 100 normal human kidneys. Papillae were dissected and digested in Cellgro containing 400 U/ml collagenase.
Cells were plated on fibronectin-coated culture flasks at a density of 104 live cells/ml in Cellgro supplemented with insulin and 10% fetal bovine serum. Confluent monolayers, which were able to withstand
600 mOsm for 8 h, were obtained within 10 to 15 d. Cells of primary isolates and first passages exhibited epithelial cell ultrastructure
including cell junctions, microvilli, and cilia. A dark-brown reaction product was observed in these cells when stained by
the immunoperoxidase method with peroxidase-labeled peanut lectin (Arachis hypogaea), which binds specifically to human distal tubule and collecting duct cells. These cells were negative for Factor-VIII (a
marker for endothelial cells) and γ-glutamyltransferase (a marker for proximal tubule cells). High activities of the glycolytic
enzyme pyruvate kinase and arginine vasopressin-stimulated cAMP production in these cells are consistent with a distal nephron
origin. The results indicate that human collecting duct cells can be isolated and cultured to provide an in vitro system to
probe pathogenetic mechanisms of potential nephrotoxins.
Part of this work was presented at a Symposium of the Center for Alternatives to Animal Testing, April 4–5, 1989, Johns Hopkins
Medical Institutions, Baltimore, MD 21205.
This work was supported in part by grants R01-AI24179, PO1-A804393 for the Public Health Service, U.S. Department of Health
and Human Services, and by a grant from the National Kidney Foundation, Baltimore, MD affiliate. 相似文献
312.
Cellular immunity elicited by human immunodeficiency virus type 1/ simian immunodeficiency virus DNA vaccination does not augment the sterile protection afforded by passive infusion of neutralizing antibodies 下载免费PDF全文
Mascola JR Lewis MG VanCott TC Stiegler G Katinger H Seaman M Beaudry K Barouch DH Korioth-Schmitz B Krivulka G Sambor A Welcher B Douek DC Montefiori DC Shiver JW Poignard P Burton DR Letvin NL 《Journal of virology》2003,77(19):10348-10356
High levels of infused anti-human immunodeficiency virus type 1 (HIV-1) neutralizing monoclonal antibodies (MAbs) can completely protect macaque monkeys against mucosal chimeric simian-human immunodeficiency virus (SHIV) infection. Antibody levels below the protective threshold do not prevent infection but can substantially reduce plasma viremia. To assess if HIV-1/SIV-specific cellular immunity could combine with antibodies to produce sterile protection, we studied the effect of a suboptimal infusion of anti-HIV-1 neutralizing antibodies in macaques with active cellular immunity induced by interleukin-2 (IL-2)-adjuvanted DNA immunization. Twenty female macaques were divided into four groups: (i). DNA immunization plus irrelevant antibody, (ii). DNA immunization plus infusion of neutralizing MAbs 2F5 and 2G12, (iii). sham DNA plus 2F5 and 2G12, and (iv). sham DNA plus irrelevant antibody. DNA-immunized monkeys developed CD4 and CD8 T-cell responses as measured by epitope-specific tetramer staining and by pooled peptide ELISPOT assays for gamma interferon-secreting cells. After vaginal challenge, DNA-immunized animals that received irrelevant antibody became SHIV infected but displayed lower plasma viremia than control animals. Complete protection against SHIV challenge occurred in three animals that received sham DNA plus MAbs 2F5 and 2G12 and in two animals that received the DNA vaccine plus MAbs 2F5 and 2G12. Thus, although DNA immunization produced robust HIV-specific T-cell responses, we were unable to demonstrate that these responses contributed to the sterile protection mediated by passive infusion of neutralizing antibodies. These data suggest that although effector T cells can limit viral replication, they are not able to assist humoral immunity to prevent the establishment of initial infection. 相似文献
313.
Selection of low-molecular-mass trypsin and chymotrypsin inhibitors based on the binding loop of CMTI-III using combinatorial chemistry methods 总被引:1,自引:0,他引:1
Kaźmierczak K Zabłotna E Jaśkiewicz A Miecznikowska H Rolka K 《Biochemical and biophysical research communications》2003,310(3):811-815
Using a combinatorial chemistry approach, a decapaptide library containing the N-terminal fragment of trypsin inhibitor CMTI-III was synthesized by the solid-phase method. The peptide library was screened for trypsin and chymotrypsin inhibitory activity applying the iterative method in solution. Two decapeptides were selected and resynthesized for each enzyme. The association equilibrium constants ((1.1+/-0.2)x10(8) and (7.3+/-1.6)x10(7)) determined for peptides with trypsin inhibitory activity indicate that they are 3-4-fold less active than the CMTI inhibitors. On the other hand, they are significantly more effective as compared with the starting sequence. Two peptides selected as chymotrypsin inhibitors displayed about 10 times higher activity (1.7+/-0.4)x10(7) and (1.1+/-0.2)x10(7), respectively) than those monosubstituted in position P(1) of the CMTI-III analogue. Considering low molecular weight of peptides selected and the lack of conformational constraints in their structures, the results are promising. They are good templates as starting sequences for further selection of small, peptidomimetic proteinase inhibitors. 相似文献
314.
Pluripotency of embryonic stem cells 总被引:2,自引:0,他引:2
Yamanaka S Li J Kania G Elliott S Wersto RP Van Eyk J Wobus AM Boheler KR 《Cell and tissue research》2008,331(1):5-22
315.
316.
Anna Adamska‐Bartłomiejczyk Attila Borics Csaba Tömböly Szabolcs Dvorácskó Marek Lisowski Alicja Kluczyk Grzegorz Wołczański Justyna Piekielna‐Ciesielska Anna Janecka 《Journal of peptide science》2017,23(12):864-870
Three novel morphiceptin analogs, in which Pro in position 2 and/or 4 was replaced by cis‐4‐aminoproline connected with the preceding amino acid through the primary amino group, were synthesized. The opioid receptor affinities, functional assay results, enzymatic degradation studies and experimental and in silico structural analysis of such analogs are presented. Copyright © 2017 European Peptide Society and John Wiley & Sons, Ltd. 相似文献
317.
Jesper H. Andersen Jacob Carstensen Daniel J. Conley Karsten Dromph Vivi Fleming‐Lehtinen Bo G. Gustafsson Alf B. Josefson Alf Norkko Anna Villnäs Ciarán Murray 《Biological reviews of the Cambridge Philosophical Society》2017,92(1):135-149
Much of the Baltic Sea is currently classified as ‘affected by eutrophication’. The causes for this are twofold. First, current levels of nutrient inputs (nitrogen and phosphorus) from human activities exceed the natural processing capacity with an accumulation of nutrients in the Baltic Sea over the last 50–100 years. Secondly, the Baltic Sea is naturally susceptible to nutrient enrichment due to a combination of long retention times and stratification restricting ventilation of deep waters. Here, based on a unique data set collated from research activities and long‐term monitoring programs, we report on the temporal and spatial trends of eutrophication status for the open Baltic Sea over a 112‐year period using the HELCOM Eutrophication Assessment Tool (HEAT 3.0). Further, we analyse variation in the confidence of the eutrophication status assessment based on a systematic quantitative approach using coefficients of variation in the observations. The classifications in our assessment indicate that the first signs of eutrophication emerged in the mid‐1950s and the central parts of the Baltic Sea changed from being unaffected by eutrophication to being affected. We document improvements in eutrophication status that are direct consequences of long‐term efforts to reduce the inputs of nutrients. The reductions in both nitrogen and phosphorus loads have led to large‐scale alleviation of eutrophication and to a healthier Baltic Sea. Reduced confidence in our assessment is seen more recently due to reductions in the scope of monitoring programs. Our study sets a baseline for implementation of the ecosystem‐based management strategies and policies currently in place including the EU Marine Strategy Framework Directives and the HELCOM Baltic Sea Action Plan. 相似文献
318.
Eric Akoko Eliot A. Atekwana Anna M. Cruse Loago Molwalefhe Wellington R. L. Masamba 《Biogeochemistry》2013,114(1-3):359-380
The Okavango River, in semi-arid northwestern Botswana, flows for over 400 km in a pristine wetland developed on a large (>22,000 km2) alluvial fan (Okavango Delta). An annual flood pulse inundates the floodplains of the wetlands and travels across the Delta in 4–6 months. In this study, we assess the effects of long hydraulic residence time, variable hydrologic interaction between river–floodplain–wetland and evapotranspiration on carbon cycling. We measured dissolved inorganic carbon (DIC) concentrations and stable carbon isotopes of DIC (δ13CDIC) from river water when the Delta was not flooded (low water) and during flooding (high water). During low water, the average DIC concentration was 31 % higher and the δ13CDIC 2.1 ‰ more enriched compared to high water. In the lower Delta with seasonally flooded wetlands, the average DIC concentration increased by 70 % during low water and by 331 % during high water compared to the Panhandle with permanently flooded wetlands. The increasing DIC concentration downriver is mostly due to evapoconcentration from transpiration and evaporation with increased transit time. The average δ13CDIC between low and high water decreased by 3.7 ‰ in the permanently flooded reaches compared to an increase of 1.6 ‰ in the seasonally flooded reaches. The lower δ13CDIC during high water in the permanently flooded reaches suggest that DIC influx from the floodplain-wetland affects river’s DIC cycling. In contrast, higher river channel elevations relative to the wetlands along seasonal flooded reaches limit hydrologic interaction and DIC cycling occurs mostly by water column processes and river-atmospheric exchange. We conclude that river-wetlands interaction and evapoconcentration are important factors controlling carbon cycling in the Okavango Delta. 相似文献
319.
Felicia Carotenuto Laura Teodori Anna Maria Maccari Luciano Delbono Giuseppe Orlando Paolo Di Nardo 《Journal of cellular and molecular medicine》2020,24(5):2704-2716
Regenerative therapies including stem cell treatments hold promise to allow curing patients affected by severe cardiac muscle diseases. However, the clinical efficacy of stem cell therapy remains elusive, so far. The two key roadblocks that still need to be overcome are the poor cell engraftment into the injured myocardium and the limited knowledge of the ideal mixture of bioactive factors to be locally delivered for restoring heart function. Thus, therapeutic strategies for cardiac repair are directed to increase the retention and functional integration of transplanted cells in the damaged myocardium or to enhance the endogenous repair mechanisms through cell-free therapies. In this context, biomaterial-based technologies and tissue engineering approaches have the potential to dramatically impact cardiac translational medicine. This review intends to offer some consideration on the cell-based and cell-free cardiac therapies, their limitations and the possible future developments. 相似文献
320.
Sylvia Ullrich Anna Münch Stephanie Neumann Elisabeth Kremmer J?rg Tatzelt Stefan F. Lichtenthaler 《The Journal of biological chemistry》2010,285(27):20664-20674
Ectodomain shedding of the amyloid precursor protein (APP) by the two proteases α- and β-secretase is a key regulatory event in the generation of the Alzheimer disease amyloid β peptide (Aβ). At present, little is known about the cellular mechanisms that control APP shedding and Aβ generation. Here, we identified a novel protein, transmembrane protein 59 (TMEM59), as a new modulator of APP shedding. TMEM59 was found to be a ubiquitously expressed, Golgi-localized protein. TMEM59 transfection inhibited complex N- and O-glycosylation of APP in cultured cells. Additionally, TMEM59 induced APP retention in the Golgi and inhibited Aβ generation as well as APP cleavage by α- and β-secretase cleavage, which occur at the plasma membrane and in the endosomes, respectively. Moreover, TMEM59 inhibited the complex N-glycosylation of the prion protein, suggesting a more general modulation of Golgi glycosylation reactions. Importantly, TMEM59 did not affect the secretion of soluble proteins or the α-secretase like shedding of tumor necrosis factor α, demonstrating that TMEM59 did not disturb the general Golgi function. The phenotype of TMEM59 transfection on APP glycosylation and shedding was similar to the one observed in cells lacking conserved oligomeric Golgi (COG) proteins COG1 and COG2. Both proteins are required for normal localization and activity of Golgi glycosylation enzymes. In summary, this study shows that TMEM59 expression modulates complex N- and O-glycosylation and suggests that TMEM59 affects APP shedding by reducing access of APP to the cellular compartments, where it is normally cleaved by α- and β-secretase. 相似文献