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Caco 2 PC12

Abigail K Elias, Denis Scanlon, Ian F Musgrave, John A Carver
Semen-derived enhancer of viral infection (SEVI) is the term given to the amyloid fibrils formed by a 39-amino acid fragment (PAP248-286) of prostatic acidic phosphatase (PAP) found in human semen. SEVI enhances human immunodeficiency virus (HIV) infectivity by four to five orders of magnitude (Münch et al., 2007). Here, we show by various biophysical techniques including Thioflavin T fluorescence, circular dichroism spectroscopy and transmission electron microscopy that fragments encompassing the central region of SEVI, i...
September 2014: Biochimica et Biophysica Acta
Cathy W C Bi, Wilson K W Luk, María-Letizia Campanari, Yuen H Liu, Li Xu, Kei M Lau, Miranda L Xu, Roy C Y Choi, Javier Sáez-Valero, Karl W K Tsim
Acetylcholinesterase (AChE) is encoded by a single gene, and the alternative splicing at the 3' end produces different isoforms, including tailed (AChET), read-through (AChER), and hydrophobic (AChEH). Different forms of this enzyme exist in different cell types. Each AChE form has been proposed to have unique function, and all of them could be found in same cell type. Thus, the splicing process of different AChE forms remains unclear. Here, we aimed to establish a quantification method in measuring the absolute amount of each AChE splicing variants within a cell type...
July 2014: Journal of Molecular Neuroscience: MN
Wei Zhou, Kun Ming Qin, Jin Jun Shan, Wen Zheng Ju, Shi Jia Liu, Bao Chang Cai, Liu Qing Di
Forsythoside A (FTA), one of the main active ingredients in weeping forsythia extract, possesses strong antibacterial, antioxidant and antiviral effects, and its content was about 8% of totally, higher largely than that of other ingredients, but the absolute bioavailability orally was approximately 0.5%, which is significant low influencing clinical efficacies of its oral preparations. In the present study, in vitro Caco-2 cell, in situ single-pass intestinal perfusion and in vivo pharmacokinetics study were performed to investigate the effects of absorption enhancers based on tight junctions: sodium caprate and water-soluble chitosan on the intestinal absorption of FTA, and the eventual mucosal epithelial damage resulted from absorption enhancers was evaluated by MTT test, measurement of total amount of protein and the activity of LDH and morphology observation, respectively...
December 15, 2012: Phytomedicine: International Journal of Phytotherapy and Phytopharmacology
Bing-Liang Ma, Yue-Ming Ma, Rong Shi, Tian-Ming Wang, Ning Zhang, Chang-Hong Wang, Yang Yang
AIM OF THE STUDY: Rhizoma Coptidis (Huanglian) is a widely used Traditional Chinese Medicine. However, it causes human as well as animal toxicities. In this study, we aimed to ascertain the toxic constituents in Rhizoma Coptidis. MATERIALS AND METHODS: The acute toxicity of both the total extract and the alkaloid-rich extract of Rhizoma Coptidis were tested in mice. The dose related tissue concentration of the Rhizoma Coptidis alkaloids in mice was determined using high performance liquid chromatography with ultraviolet detection...
March 24, 2010: Journal of Ethnopharmacology
H Satsu, T Yokoyama, N Ogawa, Y Fujiwara-Hatano, M Shimizu
The interaction between intestinal epithelial cells andperipheral neuronal cells were examined using an invitro coculture system. Two cell lines, Caco-2 and PC12, were usedfor this experiment as an intestinal epithelial and entericneuronal cell model, respectively. By coculturing with fullydifferentiated Caco-2 cells, the neurite outgrowth was inducedin PC12 cells. This neurite outgrowth in PC12 was blocked byanti-nerve growth factor (NGF) polyclonal antibodies,suggesting that the neurite outgrowth in PC12 during thecoculture with Caco-2 cells was due to NGF secreted fromCaco-2 cells...
January 2001: Cytotechnology
Cameron P Bracken, Anthony O Fedele, Sarah Linke, Wiltiana Balrak, Karolina Lisy, Murray L Whitelaw, Daniel J Peet
The hypoxia-inducible factor (HIF)-1alpha and HIF-2alpha are closely related, key transcriptional regulators of the hypoxic response, countering a low oxygen situation with the up-regulation of target genes associated with numerous processes, including vascularization and glycolysis. This involves a dual mechanism of control through both stabilization and transactivation, regulated via prolyl and asparaginyl hydroxylation. Despite high similarity with respect to protein sequence and activation pathway, a growing number of physiological and mechanistic differences between HIF-1alpha and HIF-2alpha are being reported...
August 11, 2006: Journal of Biological Chemistry
Shiro Okumura, Hiroyuki Saitoh, Tomoyuki Ishikawa, Naoya Wasano, Satoko Yamashita, Ken-Ichi Kusumoto, Tetsuyuki Akao, Eiichi Mizuki, Michio Ohba, Kuniyo Inouye
Parasporal inclusion proteins produced by Bacillus thuringiensis strain A1470 exhibit strong cytotoxicity against human leukemic T cells when activated by protease treatment. One of the cytotoxic proteins was separated by anion exchange chromatography and gel filtration chromatography and designated Cry45Aa. Its gene was then expressed in recombinant Escherichia coli, in which the Cry45Aa precursor was accumulated in an inclusion body. It was solubilized in sodium carbonate buffer and processed with proteinase K, and cytotoxic activities of the protein against various mammalian cell lines were evaluated using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide assay...
August 10, 2005: Journal of Agricultural and Food Chemistry
Hiroko Isoda, Terence P N Talorete, Junkyu Han, Shuichi Oka, Yukuo Abe, Yuhei Inamori
Organophosphorous pesticides are currently widely used in China to help boost agricultural production. However, these pesticides pose various threats to organisms, including humans, and are thus a cause of concern. Five organophosphorous pesticides, monocrotophos, omethoate, parathion-methyl, phoxim and dichlorvos, were examined for their effects on mammalian cell lines to determine their potential impact on physiological functions in vivo. Results show an increased proliferation of MCF-7 cells treated with 0...
2005: Environmental Sciences: An International Journal of Environmental Physiology and Toxicology
G Ronzitti, F Callegari, C Malaguti, G P Rossini
Yessotoxins (YTXs) are algal toxins that can be accumulated in edible molluscs. YTX treatment of MCF-7 breast cancer cells causes the accumulation of a 100 kDa fragment of E-cadherin, which we have named ECRA(100). A relative decrease in the concentrations of intact E-cadherin did not accompany the accumulation of ECRA(100) in cytosoluble extracts of MCF-7 cells on the first day of YTX treatment, but a collapse of the E-cadherin system was detected after 2-5 days of treatment with the toxin. An analysis of the general structure of ECRA(100) revealed that it consists of an E-cadherin fragment lacking the intracellular domain of the protein...
March 8, 2004: British Journal of Cancer
Gian Luigi Russo, Elio De Nisco, Gabriella Fiore, Paola Di Donato, Marco d'Ischia, Anna Palumbo
The melanin-free ink of the cephalopod Sepia officinalis is shown to contain a heat labile proteinaceous component toxic to a variety of cell lines, including PC12 cells. Gel filtration chromatography indicated that the toxic component was concentrated in those fractions eluted at a molecular weight higher than 100 kDa and exhibiting the highest tyrosinase activity. SDS-PAGE analysis of the active fractions displayed a single major band migrating at an approximate molecular weight of 100 kDa, identical with that of the single tyrosinase band in the melanin-free ink...
August 22, 2003: Biochemical and Biophysical Research Communications
Hideo Satsu, Tatsuya Yokoyama, Nobumasa Ogawa, Yoko Fujiwara-Hatano, Makoto Shimizu
The effect of neuronal cells on the functional properties of intestinal epithelial cells was examined by using an in vitro coculture system. Two cell lines, Caco-2 and PC12, were respectively used as intestinal epithelial and enteric neuronal cell models. Coculture of differentiated Caco-2 cells with PC12 caused a significant decrease in the transepithelial electrical resistance (TER) value of the Caco-2 monolayer. The permeability to lucifer yellow (LY) was also significantly increased, suggesting that the tight junction (TJ) of the Caco-2 monolayers was modulated by coculturing with PC12...
June 2003: Bioscience, Biotechnology, and Biochemistry
Michael J Haas, Shant A Parseghian, Raj M Sajid, Arshag D Mooradian
The thyroid hormone responsive protein (THRP) is a novel gene product that remains responsive to thyroid hormone in the cerebral cortex of adult rats. The biological effects of THRP are currently unknown. Since thyroid hormones (TH) are known to cause cell death in primary neuronal cultures, the effect of exogenous THRP expression on PC12 cell viability was investigated. Co-transfection of the THRP expression plasmid with the selectable marker pSV2neo resulted in a lower number of surviving PC12 cells compared to transfection with pSV2neo and the empty vector, pSVL...
May 2003: Experimental Brain Research. Experimentelle Hirnforschung. Expérimentation Cérébrale
Hongju Wu, Joanne E Nash, Pedro Zamorano, Craig C Garner
SAP97 is a modular protein composed of three PDZ domains, an SH3 domain, and a guanylate kinase-like domain. It has been implicated functionally in the assembly and structural stability of synaptic junctions as well as in the trafficking, recruitment, and localization of specific ion channels and neurotransmitter receptors. The N terminus of SAP97 (S97N) has been shown to play a key role in the selection of binding partners and the localization of SAP97 at adhesion sites, as well as the clustering of ion channels in heterologous cells...
August 23, 2002: Journal of Biological Chemistry
Serge Moskalenko, Dale O Henry, Carine Rosse, Gladys Mirey, Jacques H Camonis, Michael A White
Delivery of cytoplasmic vesicles to discrete plasma-membrane domains is critical for establishing and maintaining cell polarity, neurite differentiation and regulated exocytosis. The exocyst is a multisubunit complex required for vectorial targeting of a subset of secretory vesicles. Mechanisms that regulate the activity of this complex in mammals are unknown. Here we show that Sec5, an integral component of the exocyst, is a direct target for activated Ral GTPases. Ral GTPases regulate targeting of basolateral proteins in epithelial cells, secretagogue-dependent exocytosis in neuroendocrine cells and assembly of exocyst complexes...
January 2002: Nature Cell Biology
T Yanagiya, N Imura, S Enomoto, Y Kondo, S Himeno
Cadmium is a hazardous heavy metal existing ubiquitously in the environment, but the mechanism of cadmium transport into mammalian cells has been poorly understood. Recently, we have established a cadmium-resistant cell line (Cd-rB5) from immortalized metallothionein-null mouse cells, and found that Cd-rB5 cells exhibited a marked decrease in cadmium uptake. To investigate the mechanism of altered uptake of cadmium in Cd-rB5 cells, incorporation of various metals was determined simultaneously using a multitracer technique...
March 2000: Journal of Pharmacology and Experimental Therapeutics
C Mertens, C Kuhn, W W Franke
Using antibodies and recombinant DNA techniques, we have identified plakophilin 2, a novel desmosomal plaque protein of M(r) 100,000 (estimated from SDS-PAGE), which is a member of the arm-repeat family of proteins and can occur in two splice forms (2a and 2b) because of the insertion of a 44 amino acid (aa)-encoding exon. In its aa sequence (837 and 881 aa, calculated pIs: 9.33 and 9.38, mol wts 92,750 and 97,410 kD), it is conspicuously related to the 80-kD plakophilin 1, with which it shares a central region of 9 repeats of the arm-motif, preceeded by a long head region and followed by a very short (11 aa) carboxy-terminal sequence...
November 1996: Journal of Cell Biology
G W Varilek, G A Neil, W P Bishop, J Lin, N J Pantazis
Nerve growth factor (NGF) exists in the gut of adult rats. The cells responsible for NGF synthesis in the gut remain unknown. IEC-6 and Caco-2 cells, established cell culture models of intestinal epithelial cells, were studied to determine whether intestinal epithelial cells, were studied to determine whether they synthesize and release NGF. Conditioned media from both IEC-6 and Caco-2 cells stimulated neurite outgrowth in both rat pheochromocytoma (PC-12) cells and sensory neurons derived from embryonic chick dorsal root ganglia (DRG)...
September 1995: American Journal of Physiology
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