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991.
992.
Liberman C Takagi M Cabrera-Crespo J Sbrogio-Almeida ME Dias WO Leite LC Gonçalves VM 《Journal of industrial microbiology & biotechnology》2008,35(11):1441-1445
The high cost of the available pneumococcal conjugated vaccines has been an obstacle in implementing vaccination programs
for children in developing countries. As an alternative, Malley et al. proposed a vaccine consisting of inactivated whole-cells
of unencapsulated S. pneumoniae, which provides serotype-independent protection and involves lower production costs. Although the pneumococcus has been extensively
studied, little research has focused on its large-scale culture, thus implying a lack of knowledge of process parameters,
which in turn are essential for its successful industrial production. The strain Rx1Al− eryR was originally cultured in Todd–Hewitt medium (THY), which is normally used for pneumococcus isolation, but is unsuitable
for human vaccine preparations. The purposes of this study were to compare the strains Rx1Al− eryR and kanR, develop a new medium, and generate new data parameters for scaling-up the process. In static flasks, cell densities were
higher for eryR than kanR. In contrast, the optical density (OD) of the former decreased immediately after reaching the stationary phase, and the OD
of the latter remained stable. The strain Rx1Al− kanR was cultivated in bioreactors with medium based on either acid-hydrolyzed casein (AHC) or enzymatically hydrolyzed soybean
meal (EHS). Biomass production in EHS was 2.5 times higher than in AHC, and about ten times higher than in THY. The process
developed for growing the strain Rx1Al− kanR in pH-controlled bioreactors was shown to be satisfactory to this fastidious bacterium. The new culture conditions using
this animal-free medium may allow the production of the pneumococcal whole-cell vaccine. 相似文献
993.
Augustyniak M Babczyńska A Migula P Wilczek G Łaszczyca P Kafel A Augustyniak M 《Comparative biochemistry and physiology. Toxicology & pharmacology : CBP》2005,141(4):412-419
We studied how an exposure to an additional stressing factor-dimethoate, might affect detoxifying ability of grasshoppers collected at 5 meadow sites located along a heavy metal pollution gradient. Activities of esterases and enzymes linked with glutathione (GSH) metabolism were assayed 24 h after topical treatment with 0.32 microg dimethoate per insect. Inhibition of acetylcholinesterase (AChE) reaches nearly 50% of the value stated in untreated insects, without significant site-dependent differences. The pesticide also caused a significant decrease in activities of glutathione peroxidase (GPx) followed by a decrease in GSH levels in grasshoppers from all assayed groups, demonstrating high sensitivity of glutathione-dependent metabolism to the additional stressing factor. In the case of glutathione reductase (GR) and carboxylesterases (CarE) the fall of activity was shown especially in insects from less polluted meadows and the reference site. Glutathione reductase (GR) activity in individuals treated with dimethoate did not decrease only in insects from the most contaminated site I. This might suggest the trade-off mechanisms adapting grasshoppers to life in seriously polluted environments. 相似文献
994.
Different signaling in pig anterior pituitary cells by GnRH and its complexes with copper and nickel
Kochman K Blitek A Kaczmarek M Gajewska A Siawrys G Counis R Ziecik AJ 《Neuro endocrinology letters》2005,26(4):377-382
Gonadotropin releasing hormone (GnRH) is an essential factor in the regulation of synthesis and release of pituitary gonadotropins. After binding to specific receptors and coupling with G proteins, it triggers the intracellular signaling involving the synthesis of inositol phosphates and diacylglycerol. Previously we have showed that certain metal complexes with GnRH, i.e. copper (Cu-GnRH) and nickel (Ni-GnRH) are able to bind to the GnRH receptors. The intracellular signalling of these complexes, however, has not been yet elucidated. In this experiment, the ability of the Cu-GnRH and Ni-GnRH complexes to modulate cAMP synthesis and phosphoinositols formation in the pig anterior pituitary cells in vitro was studied. The native GnRH and its metal complexes stimulated the luteinizing hormone (LH) release, but only the effect of Cu-GnRH was found to be a dose-dependent. The metal complexes did not significantly influence inositol phosphates accumulation, while their effect on cAMP synthesis was significantly more potent than that of GnRH alone. We conclude that the Cu-GnRH and Ni-GnRH complexes increase LH release in the porcine pituitary cells although their intracellular signaling is different from that of the native GnRH. It seems that metal complexes with GnRH deserve more attention in further studies. 相似文献
995.
Nakamura K Kodaka M El-Mehasseb IM Gajewska A Okuno H Ochwanowska E Witek B Kozłowski H Kochman K 《Neuro endocrinology letters》2005,26(3):247-252
MALDI-TOF mass spectrometry, 1H NMR spectrometry, the continuous variation method and molecular modeling by MM3 calculation confirmed our earlier studies showing that gonadotropin-releasing hormone (GnRH) forms complex with copper(II) ion with the binding ratio 1:1. The copper(II) complex formed at physiological pH has a square planar configuration and GnRH complexes with nickel(II) and cobalt(II) ions are less stable than that of copper(II). 相似文献
996.
Biliński T Kwolek M Sas E Krynicka M Koziol S Owsiak-Teleon A Krzepilko A Bartosz G 《BioFactors (Oxford, England)》2005,24(1-4):59-65
A novel test for the identification of genes involved in aldehyde metabolism is proposed, based on detection of altered sensitivity of the yeast to corresponding alcohols, metabolic precursors of the aldehydes. This attitude enabled to an unexpected detection increased sensitivity of mutants devoid of CuZn-superoxide dismutase (CuZnSOD) to allyl alcohol (precursor of acrolein) and nonenol. We interpret this finding as due to inactivation of some important element of aldehyde detoxification by increased flux of superoxide in DeltaCuZnSOD mutants. 相似文献
997.
Meneses-Marcel A Marrero-Ponce Y Machado-Tugores Y Montero-Torres A Pereira DM Escario JA Nogal-Ruiz JJ Ochoa C Arán VJ Martínez-Fernández AR García Sánchez RN 《Bioorganic & medicinal chemistry letters》2005,15(17):3838-3843
A computational (virtual) screening test to identify potential trichomonacidals has been developed. Molecular structures of trichomonacidal and non-trichomonacidal drugs were represented using stochastic and non-stochastic atom-based quadratic indices and a linear discrimination analysis (LDA) was trained to classify molecules regarding their antiprotozoan activity. Validation tests revealed that our LDA-QSAR models recognize at least 88.24% of trichomonacidal lead-like compounds and suggest using this methodology in virtual screening protocols. These classification functions were then applied to find new lead antitrichomonal compounds. In this connection, the biological assays of eight compounds, selected by computational screening using the present models, give good results (87.50% of good classification). In general, most of the compounds showed high activity against Trichomonas vaginalis at the concentration of 100 microg/ml and low cytotoxicity to this concentration. In particular, two heterocyclic derivatives (VA7-67 and VA7-69) maintained their efficacy at 10 microg/ml with an important trichomonacidal activity (100.00% of reduction), but it is remarkable that the compound VA7-67 did not show cytotoxic effects in macrophage cultivations. This result opens a door to a virtual study considering a higher variability of the structural core already evaluated, as well as of other chemicals not included in this study. 相似文献
998.
Castell A Johansson P Unge T Jones TA Bäckbro K 《Protein science : a publication of the Protein Society》2005,14(7):1850-1862
The Mycobacterium tuberculosis genome contains about 4000 genes, of which approximately a third code for proteins of unknown function or are classified as conserved hypothetical proteins. We have determined the three-dimensional structure of one of these, the rv0216 gene product, which has been shown to be essential for M. tuberculosis growth in vivo. The structure exhibits the greatest similarity to bacterial and eukaryotic hydratases that catalyse the R-specific hydration of 2-enoyl coenzyme A. However, only part of the catalytic machinery is conserved in Rv0216 and it showed no activity for the substrate crotonyl-CoA. The structure of Rv0216 allows us to assign new functional annotations to a family of seven other M. tuberculosis proteins, a number if which are essential for bacterial survival during infection and growth. 相似文献
999.
Meng G Grabiec A Vallon M Ebe B Hampel S Bessler W Wagner H Kirschning CJ 《The Journal of biological chemistry》2003,278(41):39822-39829
Toll-like receptors (TLRs) mediate microbial pattern recognition in vertebrates. A broad variety of agonists has been attributed to TLR2 and three TLRs, TLR4, TLR2, and TLR5, have been demonstrated to bind microbial products. Distinct agonists might interact with different subdomains of the TLR2 extracellular domain. The TLR2 extracellular domain sequence includes 10 canonical leucine-rich repeat (LRR) motifs and 8-10 additional and potentially functionally relevant LRR-like motifs. Thus, the transfection of TLR2 LRR/LRR-like motif deletion constructs in human embryonic kidney 293 cells and primary TLR2-deficient mouse fibroblasts was performed for analysis of the role of the regarding domains in specific pattern recognition. Preparations applied as agonists were highly purified soluble peptidoglycan, lipoteichoic acid, outer surface protein A from Borrelia burgdorferi, synthetic mycoplasmal macrophage-activating lipoprotein-2, tripalmitoyl-cysteinyl-seryl-(lysyl)3-lysine (P3CSK4), dipalmitoyl-CSK4 (P2-CSK4), and monopalmitoyl-CSK4 (PCSK4) as well as lipopolysaccharide and inactivated bacteria. We found that a block of the N-terminal seven LRR/LRR-like motifs was not involved in TLR2-mediated cell activation by P3CSK4 and P2CSK4 ligands mimicking triacylated and diacylated bacterial polypeptides, respectively. In contrast, the integrity of the TLR2 holoprotein was compulsory for effective cellular recognition of other TLR2 agonists applied, including PCSK4. The formation of a functionally relevant subdomain by a region including the N-terminal seven LRR/LRR-like motifs rather than by single LRRs is suggested by our results. They further imply that TLR2 contains multiple binding domains for ligands that may contribute to the characterization of its promiscuous molecular pattern recognition. 相似文献
1000.
Vannelli TA Dykman A Ortiz de Montellano PR 《The Journal of biological chemistry》2002,277(15):12824-12829
Ethionamide (ETA), a prodrug that must undergo metabolic activation to exert its cytotoxic effects, is a second line drug against tuberculosis, a disease that infects more than a third of the world's population. It has been proposed, on the basis of genetic experiments, that ETA is activated in Mycobacterium tuberculosis by the protein encoded by the gene Rv3854c (DeBarber, A. E., Mdluli, K., Bosman, M., Bekker, L.-G., and Barry, C. E., III (2000) Proc. Natl. Acad. Sci. U. S. A. 97, 9677-9682; Baulard, A. R., Betts, J. C., Engohang-Ndong, J., Quan, S., McAdam, R. A., Brennan, P. J., Locht, C., and Besra, G. S. (2000) J. Biol. Chem. 275, 28326-28331). We report here the expression, purification, and characterization of the protein encoded by this gene. Our results establish that the enzyme (EtaA) is an FAD-containing enzyme that oxidizes ETA to the corresponding S-oxide. The S-oxide, which has a similar biological activity as ETA, is further oxidized by EtaA to 2-ethyl-4-amidopyridine, presumably via the unstable doubly oxidized sulfinic acid intermediate. This flavoenzyme also oxidizes thiacetazone, thiobenzamide, and isothionicotinamide and thus is probably responsible, as suggested by the observation of crossover resistance, for the oxidative activation of other thioamide antitubercular drugs. 相似文献