Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We tested and characterized the therapeutic value of round window membrane-delivered (RWM) d-JNKI-1 peptide (Bonny et al., 2001) against sound trauma-induced hearing loss. Morphological characteristics of sound-damaged hair cell nuclei labeled by Hoechst staining show that apoptosis is the predominant mode of cell death after sound trauma. Analysis of the events occurring after sound trauma demonstrates that c-Jun N-terminal kinase (JNK)/stress-activated protein kinase activates a mitochondrial cell death pathway (i.e., activation of Bax, release of cytochrome c, activation of procaspases, and cleavage of fodrin). Fluorescein isothiocyanate (FITC)-conjugated d-JNKI-1 peptide applied onto an intact cochlear RWM diffuses through this membrane and penetrates cochlear tissues with the exception of the stria vascularis. A time sequence of fluorescence measurements demonstrates that FITC-labeled d-JNKI-1 remains in cochlear tissues for as long as 3 weeks. In addition to blocking JNK-mediated activation of a mitochondrial cell death pathway, RWM-delivered d-JNKI-1 prevents hair cell death and development of a permanent shift in hearing threshold that is caused by sound trauma in a dose-dependent manner (EC50 = 2.05 microM). The therapeutic window for protection of the cochlea from sound trauma with RWM delivery of d-JNKI-1 extended out to 12 h after sound exposure. These results show that the mitogen-activated protein kinase/JNK signaling pathway plays a crucial role in sound trauma-initiated hair cell death. Blocking this signaling pathway with RWM delivery of d-JNKI-1 may have significant therapeutic value as a therapeutic intervention to protect the human cochlea from the effects of sound trauma.
Mol Pharmacol 2007 Mar
PMID:Inhibition of the c-Jun N-terminal kinase-mediated mitochondrial cell death pathway restores auditory function in sound-exposed animals. 1713 89

The Comet assay was used to investigate the potential of the biotoxin microcystin (MC) to induce DNA damage in the freshwater zebra mussel, Dreissena polymorpha. Mussels maintained in the laboratory were fed daily, over a 21-day period, with one of four strains of the cyanobacterium, Microcystis aeruginosa. Three of the strains produced different profiles of MC toxin, while the fourth strain did not produce MCs. The mussels were sampled at 0, 7, 14, and 21 days by withdrawing haemocytes from their adductor muscle. In addition, a positive control was performed by exposing a subsample of the mussels to water containing cadmium chloride (CdCl(2)). Cell viability, measured with the Fluorescein Diacetate/Ethidium Bromide test, indicated that the MC concentrations, to which the mussels were exposed, were not cytotoxic to the haemocytes. The Comet assay performed on the haemocytes indicated that exposure to CdCl(2) produced a dose-responsive increase in DNA damage, demonstrating that mussel haemocytes were sensitive to DNA-damaging agents. DNA damage, measured as percentage tail DNA (%tDNA), was observed in mussels exposed to the three toxic Microcystis strains, but not in mussels exposed to the nontoxic strain. Toxin analysis of the cyanobacterial cultures confirmed that the three MC-producing strains exhibit different toxin profiles, with the two MC variants detected being MC-LF and MC-LR. Furthermore, the DNA damage that was observed appeared to be strain-specific, with high doses of MC-LF being associated with a higher level of genotoxicity than low concentrations of MC-LR. High levels of MC-LF also seemed to induce relatively more persistent DNA damage than small quantities of MC-LR. This study is the first to demonstrate that in vivo exposure to MC-producing strains of cyanobacteria induces DNA damage in the haemocytes of zebra mussels and confirms the sublethal toxicity of these toxins.
Environ Mol Mutagen 2007 Jan
PMID:In vivo exposure to microcystins induces DNA damage in the haemocytes of the zebra mussel, Dreissena polymorpha, as measured with the comet assay. 1716 7

Bladder cancer is one of the most common tumors of the genitourinary tract. Here, we use phage display to identify a peptide that targets bladder tumor cells. A phage library containing random peptides was screened for binding to cells from human bladder tumor xenografts. Phage clones were further selected for binding to a bladder tumor cell line in culture. Six clones displaying the consensus sequence CXNXDXR(X)/(R)C showed selective binding to cells from primary human bladder cancer tissue. Of these, the CSNRDARRC sequence was selected for further study as a synthetic peptide. Fluorescein-conjugated CSNRDARRC peptide selectively bound to frozen sections of human bladder tumor tissue, whereas only negligible binding to normal bladder tissue was observed. When the fluorescent peptide was introduced into the bladder lumen, in a carcinogen-induced rat tumor model, it selectively bound to tumor epithelium. Moreover, when the peptide was intravenously injected into the tail vein, it homed to the bladder tumor but was not detectable in normal bladder and control organs. Next, we examined whether the peptide can detect tumor cells in urine. The fluorescent peptide bound to cultured bladder tumor cells but not to other types of tumor cell lines. Moreover, it bound to urinary cells of patients with bladder cancer, while showing little binding to urinary cells of patients with inflammation or healthy individuals. The CSNRDARRC peptide may be useful as a targeting moiety for selective delivery of therapeutics and as a diagnostic probe for the detection of bladder cancer.
Mol Cancer Res 2007 Jan
PMID:Targeting bladder tumor cells in vivo and in the urine with a peptide identified by phage display. 1725 43

Critical illness myopathy (CIM) causes significant morbidity. In this study, we investigated the effect of repeated mild hypoxia on the skeletal muscle inflammation. Sprague-Dawley rats anesthetized with 2% inhaled isoflurane were divided into two groups (n = 6 each), normoxia and hypoxia (12.5% for 12 min followed by 35% for 12 min, at which point the cycle was repeated for three times). We measured the tissue oxygen tension and perfusion (simultaneously) in hind limb skeletal muscle. Inflammation in skeletal muscle was assessed by light microcopy (Hematoxylin-Eosin staining) and apoptosis (Fluorescein-FragEL DNA fragmentation detection) and expressed as percent normoxia. Compared to the control group, hypoxia significantly (P < 0.001) altered histomorphometrics. Similarly, DNA fragmentation analysis revealed that hypoxia significantly (P < 0.001) induced apoptosis. We conclude that after a mild but repeated hypoxic insult there is marked histological alterations and induced apoptosis in skeletal muscle. We postulate that variable periods of hypoxia in the critically ill may be playing a role in the etiology of CIM.
Mol Cell Biochem 2007 Aug
PMID:Periods of systemic partial hypoxia induces apoptosis and inflammation in rat skeletal muscle. 1732 3

Eosin is a probe for the Na pump nucleotide site. In contrast to previous studies examining eosin effects on Na only ATPase, we examined Na,K-ATPase- and K-activated pNPPase activity in red blood cell membranes and purified renal Na,K-ATPase. At saturating ATP (3 mM) the eosin IC(50) for Na pump inhibition was 19 microM. Increasing ATP concentrations (0.2-2.5 mM) did not overcome eosin-induced inhibition, thus eosin is a mixed-type inhibitor of ATPase activity. To test if eosin can bind to the high-affinity ATP site, purified Na,K-ATPase was labeled with 20 microM FITC. With increasing eosin concentrations (0.1 microM-10 microM) the incorporation of FITC into the ATP site significantly decreases suggesting that eosin prevents FITC reaction at the high-affinity ATP site. Eosin was a more potent inhibitor of K-activated phosphatase activity than of Na,K-ATPase activity. At 5 mM pNPP the eosin IC(50) for Na pump inhibition was 3.8+/-0.23 microM. Increasing pNPP concentrations (0.45-14.5 mM) did not overcome eosin-induced inhibition, thus eosin is a mixed-type inhibitor of pNPPase activity. These results can be fit by a model in which eosin and ATP bind only to the nucleotide site; in some pump conformations, this site is rigid and the binding is mutually exclusive and in other conformations, the site is flexible and able to accommodate both eosin and ATP (or pNPP). Interestingly, eosin inhibition of pNPPase became competitive after the addition of C(12)E(8) (0.1%) but the inhibition of ATPase remained mixed.
Blood Cells Mol Dis
PMID:Kinetic characterization of Na,K-ATPase inhibition by Eosin. 1733 59

Resveratrol (R-3), a trihydroxy trans-stilbene from grape, inhibits multistage carcinogenesis in animal models. Here we report that 3,5,4'-trimethoxystilbene (MR-3), the permethylated derivative of R-3 was more potent against the growth of human cancer cells (HT-29, PC-3, COLO 205) with estimated IC(50) values of 81.31,42.71, and 6.25 microM, respectively. We further observed that MR-3 induced apoptosis in COLO 205 cells through modulation of mitochondrial functions regulated by reactive oxygen species (ROS). ROS generation occurs in the early stages of MR-3-induced apoptosis, preceding cytochrome-c release, caspase activation, and DNA fragmentation. Significant therapeutic effects were demonstrated in vivo by treating severe combined immune deficiency (SCID) mice bearing COLO 205 tumor xenografts with MR-3 (50 mg/kg ip). Assays on DNA fragmentation and caspase activation were performed and demonstrated that apoptosis occurred in tumor tissues treated with MR-3. The appearance of apoptotic cells, as shown by Hematoxylin and Eosin (H&E) staining, and an increase in p21 and decrease in proliferating cell nuclear antigen (PCNA) protein by immuno-histochemistry were observed in tumor tissues under MR-3 treatment. Our study identifies the novel mechanisms of the antitumor effects of MR-3 and indicates that these results may have significant applications for cancer chemotherapy.
Mol Carcinog 2008 Mar
PMID:Antitumor activity of 3,5,4'-trimethoxystilbene in COLO 205 cells and xenografts in SCID mice. 1808 28

In this work the adsorption process of Fluorescein (dye with aril-methane group) as a function of pH on three different adsorbents: goethite, Co-goethite, and magnetite has been studied experimentally and theoretically. FTIR and Raman spectroscopy have been performed in an attempt to confirm the structure of surface complexes formed by sorption of the Fluorescein to different iron oxides. Typical anionic adsorption behaviour was observed for this dye onto goethite and Co-goethite whereas the adsorption level was practically constant in the range of pH studied when the adsorbent was magnetite. The diffuse layer model was employed to fit the experimental results. The surface complexes proposed from the adsorption data were in agreement with the patterns obtained from FTIR and Raman spectroscopy. The surface structure of the oxides affects the adsorption process and the final adsorbed amount at the equilibrium. Our model of diffuse double layer with the addendum of the effect of hydrophobic forces fits well the adsorption data of Fluorescein on iron oxides at different pH in the studied range. At lower pH electrostatic forces by ligand-exchange are predominant. In the range of pH 9-11 hydrophobic forces are managing the Fluorescein adsorption on the iron oxides, with the formation of outer-sphere complexes through van der Waals/hydrophobic forces. It is interesting that in the three iron oxides studied, the adsorbed amount in this range is similar.
Spectrochim Acta A Mol Biomol Spectrosc 2008 Nov 15
PMID:Removal of Fluorescein using different iron oxides as adsorbents: effect of pH. 1830 23

A pathogenesis-related (PR) protein from Theobroma cacao (TcPR-10) was identified from a cacao-Moniliophthora perniciosa interaction cDNA library. Nucleotide and amino acid sequences showed homology with other PR-10 proteins having P loop motif and Betv1 domain. Recombinant TcPR-10 showed in vitro and in vivo ribonuclease activity, and antifungal activity against the basidiomycete cacao pathogen M. perniciosa and the yeast Saccharomyces cerevisiae. Fluorescein isothiocyanate-labeled TcPR-10 was internalized by M. perniciosa hyphae and S. cerevisiae cells and inhibited growth of both fungi. Energy and temperature-dependent internalization of the TcPR-10 suggested an active importation into the fungal cells. Chronical exposure to TcPR-10 of 29 yeast mutants with single gene defects in DNA repair, general membrane transport, metal transport, and antioxidant defenses was tested. Two yeast mutants were hyperresistant compared with their respective isogenic wild type: ctr3Delta mutant, lacking the high-affinity plasma membrane copper transporter and mac1Delta, the copper-sensing transcription factor involved in regulation of high-affinity copper transport. Acute exposure of exponentially growing yeast cells revealed that TcPR-10 resistance is also enhanced in the Snq2 export permease-lacking mutant which has reduced intracellular presence of TcPR-10.
Mol Plant Microbe Interact 2009 Jan
PMID:High-affinity copper transport and Snq2 export permease of saccharomyces cerevisiae modulate cytotoxicity of PR-10 from Theobroma cacao. 1906 1

Increased expression of tissue factor (TF) has been associated with invasive forms of breast cancer. Conversely, the loss of estrogen receptor alpha (ERalpha) is associated with increased cell invasiveness. We have examined the influence of exogenous truncated recombinant TF (rTF) on ERalpha expression and cell invasiveness and investigated the mechanism of rTF signaling. The influence of rTF on ERalpha expression in MCF-7 and T47D cell lines was investigated using reverse transcription-PCR and ELISA. Cell invasion was measured using Boyden chamber-based invasion assays. Additionally, the interaction of fluorescein-labeled rTF with the surface of MCF-7 cells and particularly with beta(1)-integrin was examined. Treatment of cells with rTF resulted in the down-regulation of ERalpha mRNA and protein over 24 h, which required beta(1)-integrin and involved the mitogen-activated protein kinase pathway but did not require PAR2 activation. The addition of rTF reduced estradiol-mediated cell proliferation as well as increased cell invasiveness requiring both PAR2 and beta(1)-integrin activation. Fluorescein-labeled rTF was shown to bind to the surface of MCF-7 cells within 5 min and peaked at 15 min. The bound rTF colocalized with cellular beta(1)-integrin and was disrupted in the presence of excess unlabeled rTF and an anti-beta(1) polyclonal antibody. Finally, affinity purification of beta(1)-integrin using rTF-conjugated agarose showed a requirement for the presence of divalent cations but not factor VIIa. The results indicate that rTF is capable of down-regulating ERalpha expression in breast cancer cells, resulting in decreases in estrogen-mediated cell proliferation and increased invasiveness. Furthermore, the mechanisms by which rTF induces these changes involve both PAR2 and beta(1)-integrin.
Mol Cancer Res 2008 Dec
PMID:Influence of exogenous tissue factor on estrogen receptor alpha expression in breast cancer cells: involvement of beta1-integrin, PAR2, and mitogen-activated protein kinase activation. 1907 26

Fluorescein (HFin) emitted strong and stable room temperature phosphorescence (RTP) on filter paper after set at 50 degrees C for 10 min using Li(+) as the ion perturber. HFin existed as Fin(-) when the pH value was in the range of 5.45-7.36. Fin(-) could react with [Cu(BPY)(2)](2+) (BPY: alpha,alpha-bipyridyl) to produce ion association complex [Cu(BPY)(2)](2+).[(Fin)(2)](2-), which could enhance the RTP signal of Hfin. In the presence of bovine serum albumin (BSA), the -COOH group of Fin(-) in the [Cu(BPY)(2)](2+).[(Fin)(2)](2-) could react with the -NH(2) group of BSA to form the ion association complex [Cu(BPY)(2)](2+).[(Fin-BSA)(2)](2-), which contained -CO-NH- bond. This complex could sharply enhance the RTP signal of Hfin and the Delta I(p) was directly proportional to the content of BSA. According to the facts above, a new solid substrate-room temperature phosphorimetry (SS-RTP) for the determination of trace protein had been established using the ion association complex [Cu(BPY)(2)](2+).[(Fin)(2)](2-)as a phosphorescent probe. This method had wide linear range (0.40 x 10(-9)-280 x 10(-9)mg l(-1)), high sensitivity (the detection limit (LD) was 1.4 x 10(-10)m gl(-1)), good precision (RSD: 3.4-4.9%) and high selectivity (the allowed concentration of coexistent ions or coexistent materials was high). It had been applied to the determination of the content of protein in 10 kinds of real samples, and the result agreed well with pyrocatechol violet-Mo (VI) method (P.V.M.M.), which indicated it had high accuracy. Meanwhile, reaction mechanism for the determination of trace protein with [Cu(BPY)(2)](2+).[(Fin)(2)](2-) phosphorescent probe was also discussed. The academic thought of this research could not only be used to develop many kinds of ion association complex phosphorescent probes, but also provided a new way to promote the sensitivity of SS-RTP.
Spectrochim Acta A Mol Biomol Spectrosc 2009 Sep 01
PMID:Solid substrate-room temperature phosphorimetry for the determination of trace protein using ion association complex [Cu(BPY)2]2+ . [(Fin)2]2- as a phosphorescent probe. 1945 14


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