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71.
Tian-Yi?Chi George?G?Chen Lok-Kee?Ho Paul?BS?LaiEmail author 《Cancer cell international》2005,5(1):27
p53 is important in the development of hepatocellular carcinoma (HCC) and in therapeutic approaches, but the mechanism whereby
it inhibits HCC growth is still unclear. The aim of the present study was to establish a HCC cell system in which p53 levels
can be regulated. Full-length wild-type p53 cDNA obtained by PCR was cloned into a retroviral response vector controlled by
the tetracycline responsive element (RevTRE-p53). The regulatory vectors RevTet-Off and RevTRE-p53 were transfected into a
packaging cell line, PT67. Hep3B cells in which the p53 gene was deleted were infected with RevTet-Off viral particles from
the PT67. Three G418-resistant cell clones with high luciferase expression and low background were infected with RevTRE-p53.
By screening dozens of RevTRE-p53-infected clones with hygromycin we identified the one with the highest expression of p53
and the lowest background after doxycycline treatment. The results showed that p53 expression in this cell clone could be
simply turned on or off by removing or adding doxycycline. Furthermore, it was found that the level of p53 protein was negatively
and sensitively related to the doxycycline concentration. In conclusion, we have established a HCC cell line in which p53
expression can be switched on or off and regulated in a dose- and time-dependent manner. 相似文献
72.
We used simulated data to investigate a number of properties of maximum-
likelihood (ML) phylogenetic tree estimation for the case of four taxa.
Simulated data were generated under a broad range of conditions, including
wide variation in branch lengths, differences in the ratio of transition
and transversion substitutions, and the absence of presence of
gamma-distributed site-to-site rate variation. Data were analyzed in the ML
framework with two different substitution models, and we compared the
ability of the two models to reconstruct the correct topology. Although
both models were inconsistent for some branch-length combinations in the
presence of site-to-site variation, the models were efficient predictors of
topology under most simulation conditions. We also examined the performance
of the likelihood ratio (LR) test for significant positive interior branch
length. This test was found to be misleading under many simulation
conditions, rejecting too often under some simulation conditions. Under the
null hypothesis of zero length internal branch, LR statistics are assumed
to be asymptotically distributed chi 2(1); with limited data, the
distribution of LR statistics under the null hypothesis varies from chi
2(1).
相似文献
73.
Cavada BS Moreno FB da Rocha BA de Azevedo WF Castellón RE Goersch GV Nagano CS de Souza EP Nascimento KS Radis-Baptista G Delatorre P Leroy Y Toyama MH Pinto VP Sampaio AH Barettino D Debray H Calvete JJ Sanz L 《The FEBS journal》2006,273(17):3962-3974
Parkia platycephala lectin 2 was purified from Parkia platycephala (Leguminosae, Mimosoideae) seeds by affinity chromatography and RP-HPLC. Equilibrium sedimentation and MS showed that Parkia platycephala lectin 2 is a nonglycosylated monomeric protein of molecular mass 29 407+/-15 Da, which contains six cysteine residues engaged in the formation of three intramolecular disulfide bonds. Parkia platycephala lectin 2 agglutinated rabbit erythrocytes, and this activity was specifically inhibited by N-acetylglucosamine. In addition, Parkia platycephala lectin 2 hydrolyzed beta(1-4) glycosidic bonds linking 2-acetoamido-2-deoxy-beta-D-glucopyranose units in chitin. The full-length amino acid sequence of Parkia platycephala lectin 2, determined by N-terminal sequencing and cDNA cloning, and its three-dimensional structure, established by X-ray crystallography at 1.75 A resolution, showed that Parkia platycephala lectin 2 is homologous to endochitinases of the glycosyl hydrolase family 18, which share the (betaalpha)8 barrel topology harboring the catalytic residues Asp125, Glu127, and Tyr182. 相似文献
74.
Kyria Santiago Nascimento Francisco Lucas Faustino do Nascimento Mayara Torquato Lima Silva Camila Bezerra Nobre Cleane Gomes Moreira Luiz André Cavalcante Brizeno Edson Lopes da Ponte Ana Maria Sampaio Assreuy Benildo Sousa Cavada 《Journal of molecular recognition : JMR》2016,29(6):248-252
Andira anthelmia (tribe Dalbergieae), a plant from Brazilian Amazon, possesses a seed lectin that was purified by affinity chromatography in sepharose–mannose. This novel Dalbergieae lectin, named AAL, agglutinated rabbit erythrocytes treated with trypsin. The hemagglutinating activity of AAL was maintained after incubation at a wide range of temperature (40 to 70 °C) and pH, was shown to be dependent on divalent cations, and was inhibited by d ‐mannose and d ‐sucrose. AAL showed an electrophoretic profile in sodium dodecyl sulfate–polyacrylamide gel electrophoresis similar to other lectins of the tribe Dalbergieae, presenting a double band of molecular weight with approximately 20 kDa and other minor bands of 17, 15, and 13 kDa, being the smaller fragment glycosylated. AAL injected by intravenous route in mice showed antinociceptive activity in two behavioral tests (writhing and formalin). In the writhing test induced by acetic acid, AAL showed inhibitory effect at 0.01 mg/kg (68%), 0.1 mg/kg (46%) and 1 mg/kg (74%). In the formalin test, AAL (0.1 mg/kg) inhibited by 48% the licking time in the inflammatory phase, an effect that was recovered by the lectin association with mannose. In conclusion, AAL presents analgesic effect involving the lectin domain via peripheral mechanisms of inflammatory nociception. This activity highlights the importance of lectins as tools to be used for understanding the interaction of protein–carbohydrate in processes associated to inflammatory pain. Copyright © 2015 John Wiley & Sons, Ltd. 相似文献
75.
Martínez CR Chanway CP Albertini AV Figueiredo MV Sampaio AH Castellon RR Cavada BS Lima-Filho JL 《FEMS microbiology letters》2004,238(1):17-22
We studied the effect of a lectin isolated from seeds of the legume Vatairea macrocarpa on net H+ efflux in Rhizobium tropici, a bacterium capable of nodulating legume Phaseolus vulgaris. V. macrocarpa lectin (VML) was observed to temporarily stimulate the specific net H(+) efflux in R. tropici. When VML was present at 32 microg ml(-1), with or without 2 microM galactose (Gal), a specific net efflux >2.4 pM H+(min)(-1) mg dry biomass(-1) was induced. There was no detectable net H+ efflux when bovine serum albumin (16 microg ml(-1)) was tested. Addition of 16 microgVMLml(-1) resulted in a 700% increase of the extracellular Na+ concentration. The soluble proteins in the supernatant containing VML extract indicate a maximum immobilization of +/-10 microgVMLml(-1), with a minimum of 36,600 dimers or 8500 larger aggregates of VML binding in each bacterium. Our data suggest that VML activates Rhizobium as a bioenergetic substrate molecule, resulting in potential alterations of the external bacterial membrane. 相似文献
76.
77.
Dam TK Gerken TA Cavada BS Nascimento KS Moura TR Brewer CF 《The Journal of biological chemistry》2007,282(38):28256-28263
Isothermal titration microcalorimetry (ITC) and hemagglutination inhibition measurements demonstrate that a chemically and enzymatically prepared form of porcine submaxillary mucin that possesses a molecular mass of approximately 10(6) daltons and approximately 2300 alpha-GalNAc residues (Tn-PSM) binds to the soybean agglutinin (SBA) with a K(d) of 0.2 nm, which is approximately 10(6)-fold enhanced affinity relative to GalNAcalpha1-O-Ser (Tn), the pancarcinoma carbohydrate antigen. The enzymatically derived 81 amino acid tandem repeat domain of Tn-PSM containing approximately 23 alpha-GalNAc residues binds with approximately 10(3)-fold enhanced affinity, while the enzymatically derived 38/40 amino acid cleavage product(s) of Tn-PSM containing approximately 11-12 alpha-GalNAc residues shows approximately 10(2)-fold enhanced affinity. A natural carbohydrate decorated form of PSM (Fd-PSM) containing 40% of the core 1 blood group type A tetrasaccharide, and 58% peptide-linked GalNAcalpha1-O-Ser/Thr residues, with 45% of the peptide-linked alpha-GalNAc residues linked alpha-(2,6) to N-glycolylneuraminic acid, shows approximately 10(4) enhanced affinity for SBA. Vatairea macrocarpa lectin (VML), which is also a GalNAc binding lectin, displays a similar pattern of binding to the four forms of PSM, although there are quantitative differences in its affinities as compared with SBA. The higher affinities of SBA and VML for Tn-PSM relative to Fd-PSM indicate the importance of carbohydrate composition and epitope density of mucins on their affinities for lectins. The higher affinities of SBA and VML for Tn-PSM relative to its two shorter chain analogs demonstrate that the length of a mucin polypeptide and hence total carbohydrate valence determines the affinities of the three Tn-PSM analogs. The results suggest a binding model in which lectin molecules "bind and jump" from alpha-GalNAc residue to alpha-GalNAc residue along the polypeptide chain of Tn-PSM before dissociating. The complete thermodynamic binding parameters for these mucins including their binding stoichiometries are presented. The results have important implications for the biological activities of mucins including those expressing the Tn cancer antigen. 相似文献
78.
Lewis Landsberg MD Louis J. Aronne MD Lawrence J. Beilin MB BS MD MA Valerie Burke MD Leon I. Igel MD Donald Lloyd‐Jones MD ScM James Sowers MD 《Obesity (Silver Spring, Md.)》2013,21(1):8-24
In light of the worldwide epidemic of obesity, and in recognition of hypertension as a major factor in the cardiovascular morbidity and mortality associated with obesity, The Obesity Society and The American Society of Hypertension agreed to jointly sponsor a position paper on obesity‐related hypertension to be published jointly in the journals of each society. The purpose is to inform the members of both societies, as well as practicing clinicians, with a timely review of the association between obesity and high blood pressure, the risk that this association entails, and the options for rational, evidenced‐based treatment. The position paper is divided into six sections plus a summary as follows: pathophysiology, epidemiology and cardiovascular risk, the metabolic syndrome, lifestyle management in prevention and treatment, pharmacologic treatment of hypertension in the obese, and the medical and surgical treatment of obesity in obese hypertensive patients. Obesity (2012) 相似文献
79.
Teixeira DI Melo LM Gadelha CA Cunha RM Bloch C Rádis-Baptista G Cavada BS Freitas VJ 《Genetics and molecular research : GMR》2006,5(1):79-87
Mammalian seminal plasma contains among others, proteins called spermadhesins, which are the major proteins of boar and stallion seminal plasma. These proteins appear to be involved in capacitation and sperm-egg interaction. Previously, we reported the presence of a protein related to spermadhesins in goat seminal plasma. In the present study, we have further characterized this protein, and we propose ion-exchange chromatography to isolate this seminal protein. Semen was obtained from four adult Saanen bucks. Seminal plasma was pooled, dialyzed against distilled water and freeze-dried. Lyophilized proteins were loaded onto an ion-exchange chromatography column. Dialyzed-lyophilized proteins from the main peak of DEAE-Sephacel were applied to a C2/C18 column coupled to an RP-HPLC system, and the eluted proteins were lyophilized for electrophoresis. The N-terminal was sequenced and amino acid sequence similarity was determined using CLUSTAL W. Additionally, proteins from DEAE-Sephacel chromatography step were dialyzed and submitted to a heparin-Sepharose high-performance liquid chromatography. Goat seminal plasma after ion-exchange chromatography yielded 6.47 +/- 0.63 mg (mean +/- SEM) of the major retained fraction. The protein was designated BSFP (buck seminal fluid protein). BSFP exhibited N-terminal sequence homology to boar, stallion and bull spermadhesins. BSFP showed no heparin-binding capabilities. These results together with our previous data indicate that goat seminal plasma contains a protein that is structurally related to proteins of the spermadhesin family. Finally, this protein can be efficiently isolated by ion-exchange and reverse-phase chromatography. 相似文献
80.
Antônia Sâmia F. do Nascimento Ana Cláudia S. Gondim João B. Cajazeiras Jorge Luis A. Correia Alana de F. Pires Kyria S. do Nascimento André Luis C. da Silva Celso S. Nagano Ana Maria S. Assreuy Benildo S. Cavada 《Journal of molecular recognition : JMR》2012,25(12):657-664
A lectin from seeds of Dioclea lasiocarpa (DLL) was purified in a single step by affinity chromatography in a Sephadex G‐50 column. DLL haemagglutinated rabbit erythrocytes showing stability even after 1 h of exposure to a different pH values (optimal between pH 6.0 and 8.0) but was inhibited after incubation with d ‐mannose and d ‐glucose. The pure protein possessed a molecular weight of 25 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis and 25,410Da by mass spectrometry. The results analyzed by the software SELCON 3 indicate that β‐sheet secondary structures are predominant in DLL (approximately 40.2% antiparallel β‐sheet, 4.6% parallel β‐sheet, 7.2% α‐helices, 17.3% turns, and 28.7% unordered structures). Mechanical activity of isolated aorta from rat measured by cumulative concentration curves of DLL, performed at the contraction plateau induced by phenylephrine in either endothelium‐intact or denuded aorta. DLL (IC50 = 34.12 ± 3.46 µg/ml) relaxed precontracted endothelized aortic rings by 34.61 ± 9.06%, 55.19 ± 11.9%, and 81.33 ± 14.35%, respectively, at 10 µg/ml (initial concentration), 30 µg/ml, and 100 µg/ml (maximum effect). All effects occurred via interaction with lectin domains and participation of nitric oxide. Copyright © 2012 John Wiley & Sons, Ltd. 相似文献