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341.
Rodrigo Lira de Oliveira Marcos Fellipe da Silva Attilio Converti Tatiana Souza Porto 《Biotechnology progress》2019,35(6):e2879
This study reports on the biochemical characterization as well as the kinetic and thermodynamic study of Aspergillus tamarii URM4634 β-fructofuranosidase (FFase) with transfructosylating activity. Conditions for FFase activity were optimized by means of a central composite rotational design using pH and temperature as the independent variables, while residual activity tests carried out in the temperature range of 45–65°C enabled us to investigate FFase thermostability and estimate the kinetic and thermodynamic parameters of enzyme denaturation. Optimal conditions for sucrose hydrolysis and fructosyl transfer catalyzed by crude FFase were 50°C, and pH 6.0 and 7.4, respectively. The thermodynamic properties of irreversible enzyme inactivation were found to be activation energy of 293.1 kJ mol−1, and activation enthalpy, entropy, and Gibbs free energy in the ranges 290.3–290.4 kJ mol−1, 568.7–571.0 J mol−1 K−1, and 97.9–108.8 kJ mol−1, respectively. The results obtained in this study point out satisfactory enzyme activity and thermostability at temperatures commonly used for industrial fructo-oligosaccharide (FOS) synthesis; therefore, this novel FFase appears to be a promising biocatalyst with great potential for long-term FOS synthesis and invert sugar production. To the best of our knowledge, this is the first report on kinetic and thermodynamic parameters of an A. tamarii FFase. 相似文献
342.
Thaís Helena Gasparoto Narciso Almeida Vieira Vinicius Carvalho Porto Ana Paula Campanelli Vanessa Soares Lara 《Immunity & ageing : I & A》2009,6(1):1-12
Background
Immunosenescence is an age-associated disorder occurring primarily in T cell compartments, including altered subset composition, functions, and activation. In women, evidence implicates diminished estrogen in the postmenopausal period as a contributing factor to diminished T cell responsiveness. Since hypoestrogenism is present in postmenopausal women, our objective focused on whether T cell activation, defined as signalling molecule expressions and activation, and function, identified as IL-2 production, were affected by low estrogen. 相似文献343.
Porto FB Perrault I Hicks D Rozet JM Hanoteau N Hanein S Kaplan J Sahel JA 《The journal of gene medicine》2002,4(4):390-396
Background
Leber's congenital amaurosis (LCA) encompasses the most precocious and severe forms of inherited retinal dystrophy, displaying very significant visual handicap at or soon after birth 1 . Among the currently identified mutations, alterations in the gene coding for retinal pigment epithelium 65‐kDa protein (RPE65) lead to LCA2 2 . Existing animal models for LCA2 (RPE65‐/‐ null mice 3 and naturally occurring RPE65‐/‐ Briard dogs 4 ) exhibit near normal retinal histology at birth, although no recordable photofunction can be detected. Structural degeneration in both cases occurs with delayed onset, cone death generally preceding that of rods.Methods
We obtained retinal tissue from a voluntarily aborted embryo of an LCA2 carrier in order to compare histopathology and immunohistochemistry with age‐matched normal foetal retina.Results
Compared to normal retinas, affected retina displayed cell loss and thinning of the outer nuclear (photoreceptor) layer, decreased immunoreactivity for key phototransduction proteins, and aberrant synaptic and inner retinal organisation. The gene mutation abolished detectable expression of RPE65 within the retinal pigment epithelium (RPE) of affected eyes, and ultrastructural examination revealed the presence of lipid and vesicular inclusions not seen in normal RPE. In addition, mutant eyes demonstrated thickening, detachment and collagen fibril disorganisation in the underlying Bruch's membrane, and the choroid was distended and abnormally vascularised, in comparison with controls.Conclusions
Such data contrast with the late‐onset ocular changes observed in animal models, indicating caution should be exercised when inferring human retinal pathophysiology from information based on other species. Copyright © 2002 John Wiley & Sons, Ltd.344.
Soares PA Nascimento CO Porto TS Correia MT Porto AL Carneiro-da-Cunha MG 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》2011,879(5-6):457-460
A PEG/citrate aqueous two-phase system was tested in the partition of commercial Concanavalin A (Con A) and subsequently applied to the extraction and purification of Con A from the crude extract of Canavalia ensiformis seeds. Con A was successfully extracted to the bottom phase of a system composed of 22% (w/w) PEG8000 and 12% (w/w) citrate at pH 6.0. The obtained purification factor was 11.5 without any loss in the hemagglutinating activity. The purity of extracted lectin was confirmed by SDS-PAGE analysis. 相似文献
345.
Luís C. Ricci Joo V. Comasseto Leandro H. Andrade Marina Capelari Quezia B. Cass Andr L.M. Porto 《Enzyme and microbial technology》2005,36(7):937-946
White-rot Basidiomycetes promoted the oxidation of aromatic pro-chiral sulfides into sulfoxides with good enantioselectivity, conversion and a small production of sulfones. The reactions were carried out using whole cells of Irpex lacteus, Pycnoporus sanguineus, Trichaptum byssogenum, Trametes rigida, Trametes versicolor and Trametes villosa. The enantioselectivity for all the aryl alkyl sulfoxides was in favor of (S)-enantiomers. Oxidation of (phenylpropyl)sulfide produced (S)-(phenylpropyl)sulfoxide with high enantiomeric excess (e.e. ≥99%) by all the Basidiomycetes employed. Basidiomycetes isolated from Brazilian biomes were used as biocatalysts in the oxidation reaction. 相似文献
346.
Veronica Schmitz Rhana Berto da Silva Prata Mayara Garcia de Mattos Barbosa Mayara Abud Mendes Sheila Santos Brand?o Thaís Porto Amadeu Luciana Silva Rodrigues Helen Ferreira Fabrício da Mota Ramalho Costa Jessica Brand?o dos Santos Fabiana dos Santos Pacheco Alice de Miranda Machado José Augusto da Costa Nery Mariana de Andrea Hacker Anna Maria Sales Roberta Olmo Pinheiro Euzenir Nunes Sarno 《PLoS neglected tropical diseases》2016,10(8)
Erythema Nodosum Leprosum (ENL) is an immune reaction in leprosy that aggravates the patient´s clinical condition. ENL presents systemic symptoms of an acute infectious syndrome with high leukocytosis and intense malaise clinically similar to sepsis. The treatment of ENL patients requires immunosuppression and thus needs to be early and efficient to prevent both disabilities and permanent nerve damage. Some patients experience multiple episodes of ENL and prolonged use of immunosuppressive drugs may lead to serious adverse effects. Thalidomide treatment is extremely effective at ameliorating ENL symptoms. Several mechanisms have been proposed to explain the efficacy of thalidomide in ENL, including the inhibition of TNF production. Given its teratogenicity, thalidomide is prohibitive for women of childbearing age. A rational search for molecular targets during ENL episodes is essential to better understand the disease mechanisms involved, which may also lead to the discovery of new drugs and diagnostic tests. Previous studies have demonstrated that IFN-γ and GM-CSF, involved in the induction of CD64 expression, increase during ENL. The aim of the present study was to investigate CD64 expression during ENL and whether thalidomide treatment modulated its expression. Leprosy patients were allocated to one of five groups: (1) Lepromatous leprosy, (2) Borderline leprosy, (3) Reversal reaction, (4) ENL, and (5) ENL 7 days after thalidomide treatment. The present study demonstrated that CD64 mRNA and protein were expressed in ENL lesions and that thalidomide treatment reduced CD64 expression and neutrophil infiltrates—a hallmark of ENL. We also showed that ENL blood neutrophils exclusively expressed CD64 on the cell surface and that thalidomide diminished overall expression. Patient classification based on clinical symptoms found that severe ENL presented high levels of neutrophil CD64. Collectively, these data revealed that ENL neutrophils express CD64, presumably contributing to the immunopathogenesis of the disease. 相似文献
347.
An exception to the generally conservative nature of plastid gene evolution
is the gene coding for the beta" subunit of RNA polymerase, rpoC2. Previous
work by others has shown that maize and rice have an insertion in the
coding region of rpoC2, relative to spinach and tobacco. To assess the
distribution of this extra coding sequence, we surveyed a broad
phylogenetic sample comprising 55 species from 17 angiosperm families by
using Southern hybridization. The extra coding sequence is restricted to
the grasses (Poaceae). DNA sequence analysis of 11 species from all five
subfamilies within the grass family demonstrates that the extra sequence in
the coding region of rpoC2 is a repetitive array that exhibits more than a
twofold increase in nucleotide substitution, as well as a large number of
insertion/deletion events, relative to the adjacent flanking sequences. The
structure of the array suggests that slipped-strand mispairing causes the
repeated motifs and adds to the mechanisms through which the coding
sequence of plastid genes are known to evolve. Phylogenetic analyses based
on the sequence data from grass species support several relationships
previously suggested by morphological work, but they are ambiguous about
broad relationships within the family.
相似文献
348.
Y Novik LM Ryan DG Haller R Asbury JP Dutcher A Schutt 《Cancer immunology, immunotherapy : CII》1999,16(4):261-266
The study was a Phase II randomized study to evaluate the efficacy of new agents for the treatment of advanced gastric carcinoma.
Patients were randomized to receive single agent chemotherapy with mitoxantrone, etoposide, aclacinomycin-A or spirogermanium.
The patients were stratified by prior use of chemotherapy, prior doxorubicin use and ECOG performance status. Patients with
a history of cardiac disease or prior doxorubicin exceeding a dose of 400 mg/m2 were restrictively randomized to sopirogermanium or etoposide only. One hundred and fourteen patients were registered for
the study. Among 98 evaluable patients there were only two partial responses (both in the etoposide arm), and one complete
response in the mitoxantrone arm. The median survival on the study was 3.3 months. One hundred and six patients were analyzable
for toxicity. There were four treatment-related deaths and four life-threatening toxicities. Because of low response rates
and relatively high toxicities the studied compounds were not deemed worth further investigation for advanced gastric cancer. 相似文献
349.
We review and further develop an analytical model that describes how thermodynamic constraints on the stability of the native state influence protein evolution in a site-specific manner. To this end, we represent both protein sequences and protein structures as vectors: structures are represented by the principal eigenvector (PE) of the protein contact matrix, a quantity that resembles closely the effective connectivity of each site; sequences are represented through the "interactivity" of each amino acid type, using novel parameters that are correlated with hydropathy scales. These interactivity parameters are more strongly correlated than the other hydropathy scales that we examine with: (1) the change upon mutations of the unfolding free energy of proteins with two-states thermodynamics; (2) genomic properties as the genome-size and the genome-wide GC content; (3) the main eigenvectors of the substitution matrices. The evolutionary average of the interactivity vector correlates very strongly with the PE of a protein structure. Using this result, we derive an analytic expression for site-specific distributions of amino acids across protein families in the form of Boltzmann distributions whose "inverse temperature" is a function of the PE component. We show that our predictions are in agreement with site-specific amino acid distributions obtained from the Protein Data Bank, and we determine the mutational model that best fits the observed site-specific amino acid distributions. Interestingly, the optimal model almost minimizes the rate at which deleterious mutations are eliminated by natural selection. 相似文献
350.