47 resultados para folded cascode


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This paper provides an insight to the trade-off between settling time and power consumption in regulated current mirrors as building parts in micropower current-switching D/A converters. The regulation-loop frequency characteristic is obtained and difficulties to impose a dominant-pole condition to the resulting 2nd-order system are evaluated. Raising pole frequencies in micropower circuits, while meeting consumption requirements, is basically limited by parasitic capacitances. For such cases, an alternative is to impose a twin-pole condition in which design constraints are somewhat relieved and settling slightly improved. Relationships between pole frequencies, transistor geometry and bias are established and design guidelines for regulated current mirrors founded. By placing loop-transistors in either weak or strong inversion, small (W/L) ratios are allowed and stray capacitances reduced. Simulated waveforms suggest a good agreement with theory. The proposed approach applied to the design of a micropower current-mode D/A converter improves both simulated and experimental settling performance.

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A minimalist representation of protein structures using a Go- like potential for interactions is implemented to investigate the mechanisms of the domain swapping of p13suc1, a protein that exists in two native conformations: a monomer and a domain- swapped dimer formed by the exchange of a beta- strand. Inspired by experimental studies which showed a similarity of the transition states for folding of the monomer and the dimer, in this study we justify this similarity in molecular descriptions. When intermediates are populated in the simulations, formation of a domain- swapped dimer initiates from the ensemble of unfolded monomers, given by the fact that the dimer formation occurs at the folding/ unfolding temperature of the monomer ( T-f). It is also shown that transitions, leading to a dimer, involve the presence of two intermediates, one of them has a dimeric form and the other is monomeric; the latter is much more populated than the former. However, at temperatures lower than T-f, the population of intermediates decreases. It is argued that the two folded forms may coexist in absence of intermediates at a temperature much lower than T-f. Computational simulations enable us to find a mechanism, `` lock- and- dock'', for domain swapping of p13suc1. To explore the route toward dimer formation, the folding of unstructured monomers must be retarded by first locking one of the free ends of each chain. Then, the other free termini could follow and dock at particular regions, where most intrachain contacts are formed, and thus de. ne the transition states of the dimer. The simulations also showed that a decrease in the maximum distance between monomers increased their stability, which is explained based on confinement arguments. Although the simulations are based on models extracted from the native structure of the monomer and the dimer of p13suc1, the mechanism of the domain- swapping process could be general, not only for p13suc1.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Xylella fastidiosa is an important pathogen bacterium transmitted by xylem-feedings leafhoppers that colonizes the xylem of plants and causes diseases on several important crops including citrus variegated chlorosis (CVC) in orange and lime trees. Glutathione-S-transferases (GST) form a group of multifunctional isoenzymes that catalyzes both glutathione (GSH)-dependent conjugation and reduction reactions involved in the cellular detoxification of xenobiotic and endobiotic compounds. GSTs are the major detoxification enzymes found in the intracellular space and mainly in the cytosol from prokaryotes to mammals, and may be involved in the regulation of stress-activated signals by suppressing apoptosis signal-regulating kinase 1. In this study, we describe the cloning of the glutathione-S-transferase from X. fastidiosa into pET-28a(+) vector, its expression in Escherichia coli, purification and initial structural characterization. The purification of recombinant xfGST (rxfGST) to near homogeneity was achieved using affinity chromatography and size-exclusion chromatography (SEC). SEC demonstrated that rxfGST is a homodimer in solution. The secondary and tertiary structures of recombinant protein were analyzed by circular dichroism and fluorescence spectroscopy, respectively. The enzyme was assayed for activity and the results taken together indicated that rxfGST is a stable molecule, correctly folded, and highly active. Several members of the GST family have been extensively studied. However, xfGST is part of a less-studied subfamily which yet has not been structurally and biochemically characterized. In addition, these studies should provide a useful basis for future studies and biotechnological approaches of rxfGST. (C) 2008 Elsevier B.V. All rights reserved.

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BACKGROUND. Stroma plays an essential role in glandular function in different systems. In the prostate, it is responsible for the development and maintenance of the differentiated state of the epithelium. The marked reduction in the epithelial compartment of the prostate gland following castration is followed by a similarly important reorganization of the stroma. In this work, we characterized the reorganization of collagen fibers in the ventral prostate of castrated rats. METHODS. Histochemical tests and immunohistochemistry for type I and III collagens plus confocal microscopy of triple-labeled (collagen III, actin, and DNA) tissue sections were employed. RESULTS. We showed that collagen fibers are composed of type I and type III collagens and that they are progressively concentrated around the epithelial structures (ducts and acini) and become increasingly undulated and folded. Double-labeling of collagen fibers and F-actin demonstrated that smooth muscle cells (SMC) are intimately associated with collagen fibers. CONCLUSIONS. The results demonstrated a marked reorganization of the collagen fibers, and suggest an active role of the SMC in the reorganization of the fibrillar components of the stroma. (C) 2000 Wiley-Liss, Inc.

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A low-voltage, low-power four-quadrant analog multiplier with optimized current-efficiency is presented. Its core corresponds to a pseudodifferential cascode, gain-boosting triode-transconductor. According to a low-voltage 1.2μm CMOS n-well process, operand differential-amplitudes are 1.0Vpp and 0.32Vpp for a 1.3V-supply. Common-mode voltages are properly chosen to maximize current-efficiency to 58%. Total quiescent dissipation is 260μW. A range of PSPICE simulation supports theoretical analysis. Excellent linearity is observed on dc characteristic. Assuming a ±0.5% mismatch on (W/L) and VTH THD at full-scale is 0.93% and 1.42%, for output frequencies of 1MHz and 10MHz, respectively.

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The cells of secretory region of the salivary glands of Pachycondyla (=Neoponera) villosa at the time of enzyme production presents the basal cellular membranes profusely folded and the intercellular junctional membranes present a few enlarged spaces. The rough endoplasmic reticulum and the Golgi bodies shift from being flat and small vesicular cisternae to enlarged vesicular cisternae according to the cell physiological state and characterize an asynchronic cell cycle. Enzymes are released into the lumen by microapocrine secretion. The stage of silk production is detected after a behavioral act, when the nurse worker separates the mature larva. At this time, the salivary gland cells present only one physiological state (synchronized secretory cycle): this state was characterized by basal cellular membrane poorly folded, intercellular junctions presenting some small spaces, rough endoplasmic reticulum compounded by flat cistenae, enlarged Golgi bodies with fibrous material inside and a few secretory vesicles containing silk, which undergo exocytosis. The silk in the lumen shows 2 forms: tactoid and flocculent material.

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N-Terminally and internally labeled analogues of the hormones angiotensin (AII, DRVYIHPF) and bradykinin (BK, RPPGFSPFR) were synthesized containing the paramagnetic amino acid 2,2,6,6-tetramethylpiperidine-1-oxyl-4-amino-4- carboxylic acid (TOAC). TOAC replaced Asp 1 (TOAC 1-AII) and Val 3 (TOAC 3-AII) in AII and was inserted prior to Arg 1 (TOAC 0-BK) and replacing Pro 3 (TOAC 3-BK) in BK. The peptide conformational properties were examined as a function of trifluoroethanol (TFE) content and pH. Electron paramagnetic resonance spectra were sensitive to both variables and showed that internally labeled analogues yielded rotational correlation times (TC) considerably larger than N-terminally labeled ones, evincing the greater freedom of motion of the N-terminus. In TFE, τ C increased due to viscosity effects. Calculation of τ Cpeptide/τ CTOAC ratios indicated that the peptides acquired more folded conformations. Circular dichroism spectra showed that, except for TOAC 1-AII in TFE, the N-terminally labeled analogues displayed a conformational behavior similar to that of the parent peptides. In contrast, under all conditions, the TOAC 3 derivatives acquired more restricted conformations. Fluorescence spectra of All and its derivatives were especially sensitive to the ionization of Tyr 4. Fluorescence quenching by the nitroxide moiety was much more pronounced for TOAC 3-AII The conformational behavior of the TOAC derivatives bears excellent correlation with their biological activity, since, while the N-terminally labeled peptides were partially active, their internally labeled counterparts were inactive [Nakaie, C. R., et al., Peptides 2002, 23, 65-70]. The data demonstrate that insertion of TOAC in the middle of the peptide chain induces conformational restrictions that lead to loss of backbone flexibility, not allowing the peptides to acquire their receptor-bound conformation. © 2004 Wiley Periodicals, Inc.

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Samples from 9 llamas (28 through 36 weeks of gestation) were collected and fixed in 4% buffered paraformaldehyde (light microscopy) and in 2.5% buffered glutaraldehyde (transmission and scanning electron microscopy). The material was processed in paraplast and slides (5μm) were stained with HE, PAS, Masson-Trichrome, acid phosphatase and Perl's. The uteroferrin was immunolocalized. The results show that llama placenta is chorioallantoic, diffuse, folded and epitheliochorial, and the fetus is covered with an epidermal membrane. The trophoblast cells have variable morphology: cubic, rounded and triangular cells, with cytoplasm containing PAS-positive granules. Binucleated cells with large cytoplasm and rounded nuclei, as well as giant trophoblastic cells with multiple nuclei were also observed. Numerous blood vessels were observed beneath the cells of the uterine epithelium and around the chorionic subdivided branches. Glandular activity was shown by PAS, Perl's, and acid phosphatase positive reactions in the cytoplasm and glandular lumen, and by immunolocalization of the uteroferrin in the glandular epithelium. The uterine glands open in spaces formed by the areoles, which are filled by PAS-positive material. The llama fetus was covered by the epidermal membrane, composed of stratified epithelium, with up to seven layers of mono-, bi- or trinucleated cells. The high level of maternal and fetal vascularization surfaces indicates an intense exchange of substances across both surfaces. The metabolic activity shown in the uterine glands suggests an adaptation of the gestation to the high altitudes of the natural habitat of this species.

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The Neoproterozoic Granitic Province from the southeast of Mato Grosso and Mato Grosso do Sul region is constituted by seven distinctive granitic bodies, emplaced in the epimetamorphic rocks of the Cuiabá Group and related to the evolution of Paraguai Folded Belt. The Northern portion crops out in Mato Grosso State and is constituted by São Vicente, Araguaiana and Lajinha batholiths; the Southern portion crops out in the Mato Grosso do Sul State and is represented the Sonora, Coxim Rio Negro and Taboco massifs. The structural evolution is evidenced by the presence of three deformational phases, the first one is characterized by axial planar foliation (S 1), parallel or sub parallel to S 0, with attitude N60E/70NW; the second constitutes the most important phase for the tectonic arrangement and is defined by a slaty and/or crenulation cleavages (S 2), with attitude N10W/30SW and contact metamorphism associated to the emplacement of granitic bodies, the third phase is characterized by a weak retrometamorphic cleavage (S 3) with attitude N10E/ 80NW, that evolves local and gradually to strike-slip shear zones. The geological data suggest that this province was formed syn- to post tectonic to the D2 deformational set associated to the end of the collisional event in the Southeastern of the Amazonian Craton.

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Rio Apa Massif crops out in the Mato Grosso do Sul state and corresponds to the southeastern portion of the Amazonian Craton dominantly Paleoproterozoic in age. Rio Apa Complex is oldest and it is composed mainly by migmatitic orthogneisses, beyond amphybolites, tonalities and granodiorite. Alto Tererê Group is composed by schists, biotitemuscovite gneisses and micaceous quartzites generally rich in garnets, beyond metabasic rocks of low amphibolite facies. The Amoguijá Group is constituted by Alumiador Intrusive Suite, which is represented by a sieno to monzogranitic batholith and Serra da Bocaina Volcanic Suite composed of volcanoclastic rocks of alkali riolites to monzoriolites compositions and pyroclastic products. Overlaying towards East and South occurs Neoproterozoic metasedimentary rocks from the Paraguai Folded Belt (Cuiabá, Corumbá and Jacadigo Groups - Urucum Formation). Structural-metamorphic framewok is identified by five deformational phases but the actual tectonic and metamorphic structure shows the superposed tectonic array of the Paraguai Folded Belt. Rocks from Rio Apa Complex, Alto Tererê Group and Amoguijá Group record an older structural evolution defined by (Dn-1 and Dn). The deformational phases (Dn+1 and Dn+2) are visible mainly in rocks of Paraguai Folded Belt beyond the last deformation (Dn+3) that imprints all sequences.

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Neurospora crassa has been widely used as a model organism and contributed to the development of biochemistry and molecular biology by allowing the identification of many metabolic pathways and mechanisms responsible for gene regulation. Nuclear proteins are synthesized in the cytoplasm and need to be translocated to the nucleus to exert their functions which the importin-α receptor has a key role for the classical nuclear import pathway. In an attempt to get structural information of the nuclear transport process in N. crassa, we present herein the cloning, expression, purification and structural studies with N-terminally truncated IMPα from N. crassa (IMPα-Nc). Circular dichroism analysis revealed that the IMPα-Nc obtained is correctly folded and presents a high structural conservation compared to other importins-α. Dynamic light scattering, analytical size-exclusion chromatography experiments and molecular dynamics simulations indicated that the IMPα-Nc unbound to any ligand may present low stability in solution. The IMPα-Nc theoretical model displayed high similarity of its inner concave surface, which binds the cargo proteins containing the nuclear localization sequences, among IMPα from different species. However, the presence of non-conserved amino acids relatively close to the NLS binding region may influence the binding specificity of IMPα-Nc to cargo proteins. Copyright © 2012 Bentham Science Publishers. All Rights Reserved.

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Background: Cimetidine, histamine H2 receptors antagonist, has caused adverse effects on the male hormones and reproductive tract due to its antiandrogenic effect. In the testes, peritubular myoid cells and muscle vascular cells death has been associated to seminiferous tubules and testicular microvascularization damages, respectively. Either androgen or histamine H2 receptors have been detected in the mucosa and smooth muscular layer of vas deferens. Thus, the effect of cimetidine on this androgen and histamine-dependent muscular duct was morphologically evaluated.Methods: The animals from cimetidine group (CMTG; n=5) received intraperitoneal injections of 100 mg/kg b.w. of cimetidine for 50 days; the control group (CG) received saline solution. The distal portions of vas deferens were fixed in formaldehyde and embedded in paraffin. Massońs trichrome-stained sections were subjected to morphological and the following morphometrical analyzes: epithelial perimeter and area of the smooth muscular layer. TUNEL (Terminal deoxynucleotidyl-transferase mediated dUTP Nick End Labeling) method, NF-kB (nuclear factor kappa B) and AR (androgen receptors) immunohistochemical detection were also carried out. The birefringent collagen of the muscular layer was quantified in picrosirius red-stained sections under polarized light. The muscular layer was also evaluated under Transmission Electron Microscopy (TEM).Results: In CMTG, the mucosa of vas deferens was intensely folded; the epithelial cells showed numerous pyknotic nuclei and the epithelial perimeter and the area of the muscular layer decreased significantly. Numerous TUNEL-labeled nuclei were found either in the epithelial cells, mainly basal cells, or in the smooth muscle cells which also showed typical features of apoptosis under TEM. While an enhanced NF-kB immunoexpression was found in the cytoplasm of muscle cells, a weak AR immunolabeling was detected in these cells. In CMTG, no significant difference was observed in the birefringent collagen content of the muscular layer in comparison to CG.Conclusions: Cimetidine induces significant damages in the epithelium; a possible antiandrogenic effect on the basal cells turnover should be considered. The cimetidine-induced muscle cells apoptosis confirms the susceptibility of these cells to this drug. The parallelism between enhanced cytoplasmic NF-kB immunolabeling in the damaged muscular tissue and muscle cell apoptosis suggests that this drug may avoid the translocation of NF-kB to the nucleus and interfere in the control of NF-kB-mediated smooth muscle cell apoptosis. The decreased immunoexpression of ARs verified in the damaged muscular tissue reinforces this possibility. © 2013 Koshimizu et al.; licensee BioMed Central Ltd.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)