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Vujčić M Lazić M Milenković M Sladić D Radulović S Filipović N Anđelković K 《Journal of biochemical and molecular toxicology》2011,25(3):175-182
Organometallic Cd(II) compounds have recently attracted attention for their anticancer activity. The interaction of the dinuclear complex of Cd(II) with the condensation product of 2-acetylpyridine and malonic acid dihydrazide, N',N'(2) -bis[(1E)-1-(2-pyridyl)ethylidene]propanedihydrazide (Cd(II)H(2) L), with calf thymus DNA (CT-DNA) was monitored by blue shift in UV-vis spectra of the complex. The binding constant of Cd(II)H(2) L complex with CT-DNA was determined (K(B) = 1.8 × 10(4) M(-1) ) and was indicative of minor groove binding. Agarose gel electrophoretic changes in mobility of supercoiled and circular forms of pBR322 and pUC18 plasmids in the presence of the complex suggest that conformational changes in the plasmids occur upon binding of the Cd(II)H(2) L complex. The Cd(II)H(2) L complex induced perturbation of the cell cycle phase distribution and an increase in the percentage of cells in the sub-G1 phase of human cervical cancer HeLa cell line and murine melanoma B16 cell line. Immunoblotting analysis showed the overexpression of Bcl-2 protein with the Cd(II)H(2) L complex. 相似文献
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Exploration of the function and organization of the yeast early secretory pathway through an epistatic miniarray profile 总被引:23,自引:0,他引:23
Schuldiner M Collins SR Thompson NJ Denic V Bhamidipati A Punna T Ihmels J Andrews B Boone C Greenblatt JF Weissman JS Krogan NJ 《Cell》2005,123(3):507-519
We present a strategy for generating and analyzing comprehensive genetic-interaction maps, termed E-MAPs (epistatic miniarray profiles), comprising quantitative measures of aggravating or alleviating interactions between gene pairs. Crucial to the interpretation of E-MAPs is their high-density nature made possible by focusing on logically connected gene subsets and including essential genes. Described here is the analysis of an E-MAP of genes acting in the yeast early secretory pathway. Hierarchical clustering, together with novel analytical strategies and experimental verification, revealed or clarified the role of many proteins involved in extensively studied processes such as sphingolipid metabolism and retention of HDEL proteins. At a broader level, analysis of the E-MAP delineated pathway organization and components of physical complexes and illustrated the interconnection between the various secretory processes. Extension of this strategy to other logically connected gene subsets in yeast and higher eukaryotes should provide critical insights into the functional/organizational principles of biological systems. 相似文献
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ER-associated degradation (ERAD) of glycoproteins depends on dual recognition of protein misfolding and remodeling of the substrate's N-linked glycans. After recognition, substrates are retrotranslocated to the cytosol for proteasomal degradation. To explore the directionality of this process, we fused a highly stable protein, DHFR, to the N or C terminus of the soluble ERAD substrate CPY* in yeast. Degradation of the C-terminal CPY*-DHFR fusion is markedly slowed and is accompanied by DHFR release in the ER lumen. Thus, folded lumenal domains can impede protein retrotranslocation. The ER lumenal protein Yos9p is required for both release of DHFR and degradation of multiple ERAD substrates. Yos9p forms a complex with substrates and has a sugar binding pocket that is essential for its ERAD function. Nonetheless, substrate recognition persists even when the sugar binding site is mutated or CPY* is unglycosylated. These and other considerations suggest that Yos9p plays a critical role in the bipartite recognition of terminally misfolded glycoproteins. 相似文献
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Valentina Slavevska-Stamenkovi? Momir Paunovi? Stoe Smiljkov Traj?e Stafilov Dana Preli? Milica Ristovska Zoran Ga?i? Ana Atanackovi? 《Biologia》2012,67(6):1129-1142
In the present study, we analysed spatial and temporal heterogeneity of the limnological characteristics to provide more detailed information about the processes taking place within Mantovo Reservoir (Republic of Macedonia). The relationship between principal macroinvertebrate species and environmental variables was analysed in order to explore factors that dominantly affect community distribution pattern. Unlike the most reservoirs, strong longitudinal gradient for suspended organic matter and nutrients (total phosphorous, nitrates and nitrites) along the reservoir doesn??t exist. However, the process of thermal stratification has a strong influence on the metabolism and structure of the Mantovo ecosystem, which can be demonstrated by the vertical and longitudinal distribution of dissolved oxygen (DO), CO2, pH and metals concentrations. Canonical Correspondence Analysis (CCA) indicated that the main factors controlling spatial distribution of Limnodrilus hoffmeisteri and Chironomus plumosus group were temperature, dissolved oxygen and manganese, including sulphates for C. plumous group. Chaoborus crystallinus showed opposite distribution pattern. Cladotanytarsus mancus group was strongly associated with shallower part (littoral and sublittoral) of Mantovo Reservoir characterized by favourable oxygen condition. None of the environmental variables included in CCA showed any relationship with density of Procladius sp. 相似文献
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Stojakovic M Salazar-Fontana LI Tatari-Calderone Z Badovinac VP Santori FR Kovalovsky D Sant'Angelo D Harty JT Vukmanovic S 《Journal of immunology (Baltimore, Md. : 1950)》2008,181(10):6770-6778
Central tolerance plays a significant role in preventing autoimmune diseases by eliminating T cells with high and intermediate avidity for self. To determine the manner of setting the threshold for deletion, we created a unique transgenic mouse strain with a diverse T cell population and globally increased TCR avidity for self-peptide/MHC complexes. Despite the adaptations aimed at reducing T cell reactivity (reduced TCR levels and increased levels of TCR signaling inhibitor CD5), transgenic mice displayed more severe experimental allergic encephalomyelitis and lupus. The numbers and activity of natural (CD4(+)CD25(+)) regulatory T cells were not altered. These findings demonstrate that the threshold for deletion is adaptable, allowing survival of T cells with higher avidity when TCR avidity is globally increased. 相似文献
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Nestorović N Manojlović-Stojanoski M Ristić N Sekulić M Sosić-Jurjević B Filipović B Milosević V 《Histochemistry and cell biology》2008,130(4):699-708
The effects of multiple somatostatin (SRIH-14) administration on the pituitary-ovarian axis were examined in peripubertal rats. Female Wistar rats received subcutaneously, two daily doses of 20 mug SRIH-14 per 100 g body weight (b.w.) for five consecutive days (from the 33rd to the 37th day of life). Follicle-stimulating (FSH), luteinizing (LH) and somatotropic (GH) cells were examined by the peroxidase-anti-peroxidase immunocytochemical method. Changes in cell volumes, volume densities and number per unit area (mm(2)) of FSH-, LH- and GH-immunoreactive cells were evaluated by stereology and morphometry. Serum FSH and LH levels were determined by RIA. Ovaries were analyzed by simple point counting of follicles. The volumes and volume densities of FSH-, LH- and GH-immunoreactive cells were significantly decreased while their numbers per mm(2) remained unchanged. SRIH-14 induced a significant decrease in serum FSH and LH levels. In the ovary, SRIH-14 induced an increase in the number of primordial follicles, followed by a reduction in the number of small healthy growing follicles and absence of preovulatory follicles. The number of atretic follicles was unchanged. We concluded that treatment with SRIH-14 during the peripubertal period markedly inhibited pituitary FSH, LH and GH cells. In the ovary, SRIH-14 acted by inhibiting folliculogenesis without affecting atretic processes. 相似文献
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Boris Sakic Marcella P. A. Cooper Sarah E. Taylor Milica Stojanovic Bosa Zagorac Minesh Kapadia 《Journal of visualized experiments : JoVE》2015,(98)
Due to rapid advances in genetic engineering, small rodents have become the preferred subjects in many disciplines of biomedical research. In studies of chronic CNS disorders, there is an increasing demand for murine models with high validity at the behavioral level. However, multiple pathogenic mechanisms and complex functional deficits often impose challenges to reliably measure and interpret behavior of chronically sick mice. Therefore, the assessment of peripheral pathology and a behavioral profile at several time points using a battery of tests are required. Video-tracking, behavioral spectroscopy, and remote acquisition of physiological measures are emerging technologies that allow for comprehensive, accurate, and unbiased behavioral analysis in a home-base-like setting. This report describes a refined phenotyping protocol, which includes a custom-made monitoring apparatus (Integrated Behavioral Station, INBEST) that focuses on prolonged measurements of basic functional outputs, such as spontaneous activity, food/water intake and motivated behavior in a relatively stress-free environment. Technical and conceptual improvements in INBEST design may further promote reproducibility and standardization of behavioral studies. 相似文献