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991.
Distribution of anticancer antibiotic daunomycin in the rat heart and kidney revealed by immunocytochemistry using monoclonal antibodies 总被引:3,自引:3,他引:0
Two monoclonal antibodies (ADM-1-11 and 79-31 mAbs) were raised against daunomycin (DM) conjugated to bovine serum albumin
via the cross-linker N-(gamma-maleimidobutyryloxy)succinimide. The monoclonal antibodies (mAbs) specifically detected DM as well as its analogs
doxorubicin and epirubicin, but did not react with other anticancer antibiotics, including pepleomycin, mitomycin C, and actinomycin
D. The mAbs reacted strongly with glutaraldehyde-conjugated DM in an enzyme linked immunosorbent assay (ELISA) used as a model
system for immunocytochemistry as well as in appropriately pretreated sections of tissues from animals injected with DM. No
staining occurred in tissues from uninjected animals. In order to perform DM ICC a number of tissue treatment conditions critical
to the detection of low molecular weight substances were employed. Uptake of DM was studied in rats after a single i.v. or
i.p. administration of the drug. In the heart, accumulation of DM occurred in nuclei and in the cytoplasm. In the kidney,
DM immunoreactivity accumulated in all segments of the nephron except for the proximal tubules. Since the proximal tubules
are known to be where a variety of transport systems including P-glycoprotein (Pgp) and organic anion-transporting polypeptides
(OATPs) in drug interactions occur, the absence of DM accumulation in these segments may reflect a transport phenomenon depending
upon such transporters. The availability of methods to study sites of accumulation of DM offers possibilities for understanding
toxic side effects of this drug on the heart and kidney. Moreover, the immunocytochemical methodology developed may prove
useful for the localization of other low molecular weight drugs that can be fixed in situ by glutaraldehyde. 相似文献
992.
Improved immunocytochemical detection of daunomycin 总被引:3,自引:3,他引:0
Improved immunocytochemical (ICC) detection of the anthracycline anticancer antibiotic daunomycin (DM) has been achieved by
use of hydrogen peroxide oxidation prior to ICC staining for DM. The new method greatly enhanced the localization of DM accumulation
in cardiac, smooth and skeletal muscle of rats after a single i.v. dose of the drug. DM accumulated in the nuclei as well
as in the sarcoplasm, where it occurred in the form of small granules, which were particularly evident in cardiac muscle cells.
The distribution of the granules coincided with that of mitochondria. Uptake of DM in nuclei and mitochondria of heart muscle
cells may help to improve our understanding of the cardiac toxicity of DM and related anthracyclin antibiotics. A number of
ELISA tests were carried out in order to elucidate the mechanims of H2O2−assisted antigen retrieval. A possible mechanism is that DM is reduced and converted to its semiquinone and/or hydroquinone
derivative in vivo. Oxidation by hydrogen peroxide acts to convert these derivatives back to the native antigen. The improved
ICC methodology using oxidation to recreate native antigens from reduced metabolites may be helpful also with respect to the
localization of other drugs. 相似文献
993.
Li J Brieher WM Scimone ML Kang SJ Zhu H Yin H von Andrian UH Mitchison T Yuan J 《Nature cell biology》2007,9(3):276-286
Coordinated regulation of cell migration, cytokine maturation and apoptosis is critical in inflammatory responses. Caspases, a family of cysteine proteases, are known to regulate cytokine maturation and apoptosis. Here, we show that caspase-11, a mammalian pro-inflammatory caspase, regulates cell migration during inflammation. Caspase-11-deficient lymphocytes exhibit a cell-autonomous migration defect in vitro and in vivo. We demonstrate that caspase-11 interacts physically and functionally with actin interacting protein 1 (Aip1), an activator of cofilin-mediated actin depolymerization. The caspase-recruitment domain (CARD) of caspase-11 interacts with the carboxy-terminal WD40 propeller domain of Aip1 to promote cofilin-mediated actin depolymerization. Cells with Aip1 or caspase-11 deficiency exhibit defects in actin dynamics. Using in vitro actin depolymerization assays, we found that caspase-11 and Aip1 work cooperatively to promote cofilin-mediated actin depolymerization. These data demonstrate a novel cell autonomous caspase-mediated mechanism that regulates actin dynamics and mammalian cell migration distinct from the receptor mediated Rho-Rac-Cdc42 pathway. 相似文献
994.
Directional migration is important in wound healing by epithelial cells. Recent studies have shown that polarity proteins such as mammalian Partitioning-defective 6 (Par6), atypical protein kinase C (aPKC) and mammalian Discs large 1 (Dlg1) are crucial not only for epithelial apico-basal polarity, but also for directional movement. Here, we show that the protein associated with Lin seven 1 (PALS1)-associated tight junction protein (PATJ), another evolutionarily conserved polarity protein, is also required for directional migration by using a wound-induced migration assay. In addition, we found that aPKC and Par3 localize to the leading edge during migration of epithelia and that PATJ regulates their localization. Furthermore, our results show that microtubule-organizing centre orientation is disrupted in PATJ RNA interference (RNAi) MDCKII (Madin-Darby canine kidney II) cells during migration. Together, our data indicate that PATJ controls directional migration by regulating the localization of aPKC and Par3 to the leading edge. The migration defect in PATJ RNAi cells seems to be due to the disorganization of the microtubule network induced by mislocalization of polarity proteins. 相似文献
995.
996.
Goang-Won Cho Seung Min Shin Hyun Kee Kim Seon-Ah Ha Sanghee Kim Joo-Hee Yoon Soo Young Hur Tae Eung Kim Jin Woo Kim 《BMC cell biology》2007,8(1):50
Background
The Human cervical cancer oncogene (HCCR-1) has been isolated as a human oncoprotein, and has shown strong tumorigenic features. Its potential role in tumorigenesis may result from a negative regulation of the p53 tumor suppressor gene. 相似文献997.
Sung Hun Youn Hyun Jung Kim Tae Han Kim Chul Soo Shin 《Journal of Molecular Catalysis .B, Enzymatic》2007,46(1-4):26-31
Acylation reactions of naringin with palmitic acid were performed by a lipase after formation of highly concentrated homogeneous solutions. Their initial naringin concentration was 840–950 mM, which is 20–60 times greater than that in organic solvent media. The overall productivity of highly concentrated solutions was more than 15 times greater than those of organic phase media. The addition of DMSO (20–40%, w/w) to substrate mixtures lowered the melting temperature of a naringin–palmitic acid mixture (1:1 molar ratio) to about 40 °C. Reactions at 80 °C apparently followed Michaelis–Menten kinetics despite extremely high substrate concentrations. As the temperature increased from 60 °C to 80 °C, the apparent viscosity of the highly concentrated solution decreased remarkably from 4.31 Pa s to 0.063 Pa s. An activation energy of 7.65 kcal/mol obtained in a range of 60–75 °C suggests a diffusion-control. On the other hand, an activation energy of 17.09 kcal/mol in a range of 75–90 °C indicates a reaction-control. The highest product conversion yield of 33% (mol/mol) was obtained in a 10 h reaction at 80 °C. Addition of activated molecular sieves to the highly concentrated solution increased the product conversion yield by 7% (mol/mol), suggesting that the original equilibrium was disrupted by removing water and then a new equilibrium was reached. 相似文献
998.
Kang S Kim EY Bahn YJ Chung JW Lee do H Park SG Yoon TS Park BC Bae KH 《Cellular & molecular biology letters》2007,12(1):139-147
Oxidative stress has been implicated in the pathogenesis of neuronal degenerative diseases. It is also widely known that oxidative
stress induces mitogen-activated protein kinase (MAPK) signaling cascades. In this study, we used proteomic analysis to investigate
the role of the MAPK pathway in oxidative stress-induced neuronal cell death. The results demonstrated that several proteins,
including eukaryotic translation elongation factor 2 (eEF2) and enolase I, showed a differential expression pattern during
the neuronal cell death process, and this was MAPK pathway dependent. Several chaperone and cytoskeletal proteins including
heat shock protein 70, calreticulin, vimentin, prolyl 4-hydroxylase β polypeptide, and transgelin 2 were up-or down-regulated,
despite their expressions not depending on the MAPK pathway. These findings strongly suggest that the expressions of proteins
which play protective roles are independent of the MAPK pathway. On the other hand, eEF2 and enolase I may be the downstream
targets of the MAPK pathway. 相似文献
999.
Shin M Kadowaki T Iwata J Kawakubo T Yamaguchi N Takii R Tsukuba T Yamamoto K 《Biological chemistry》2007,388(11):1173-1181
Cathepsin E (CE) is an intracellular aspartic proteinase implicated in various physiological and pathological processes, yet its actual roles in vivo remain elusive. To assess the physiological significance of CE expression in tumor cells, human CE was stably expressed in human prostate carcinoma ALVA101 cells expressing very little CE activity. Tumor growth in nude mice with xenografted ALVA101/hCE cells was slower than with control ALVA101/mock cells. Angiogenesis antibody array and ELISA assay showed that this was partly due to the increased expression of some antiangiogenic molecules including interleukin 12 and endostatin in tumors induced by CE expression. In vitro studies also demonstrated that, among the cathepsins tested, CE most efficiently generated endostatin from the non-collagenous fragment of human collagen XVIII at mild acidic pH. Histological examination revealed that tumors formed by ALVA101/hCE cells were partitioned by well-developed membranous structures and covered with thickened, well-stratified hypodermal tissues. In addition, both the number and extent of activation of tumor-infiltrating macrophages were more profound in ALVA101/hCE compared to ALVA101/mock tumors. The chemotactic response of macrophages to ALVA101/hCE cells was also higher than that to ALVA/mock cells. These results thus indicate that CE expression in tumor cells induces tumor growth arrest via inhibition of angiogenesis and enhanced immune responses. 相似文献
1000.
During endochondral ossification, a process that accounts for the majority of bone formation in vertebrates, hypertrophic chondrocytes display a greater susceptibility to apoptosis when compared to proliferating chondrocytes. However, the molecular mechanisms underlying this phenomenon remain unclear. Nkx3.2, a member of the NK class of homeoproteins, is initially expressed in chondrogenic precursor cells, and later, during cartilage maturation, its expression is restricted to proliferating chondrocytes. Here, we show that the nuclear factor kappa B (NF-kappaB) pathway is required for chondrocyte viability and that Nkx3.2 supports chondrocyte survival by constitutively activating RelA. Although signal-dependent NF-kappaB activation has been intensively studied, ligand-independent NF-kappaB activation is poorly understood. The data presented here support a novel ligand-independent mechanism of NF-kappaB activation, whereby Nkx3.2 recruits the RelA-IkappaBalpha heteromeric complex into the nucleus by direct protein-protein interactions and activates RelA through proteasome-dependent IkappaBalpha degradation in the nucleus. Furthermore, we demonstrate that stage-specific NF-kappaB activation, mediated by Nkx3.2, regulates chondrocyte viability during cartilage maturation. 相似文献