Biophysical Characterization of Interactions Involving Importin-α during Nuclear Import

Proteins containing the classical nuclear localization sequences (NLSs) are imported into the nucleus by the importin-α/β heterodimer. Importin-α contains the NLS binding site, whereas importin-β mediates the translocation through the nuclear pore. We characterized the interactions involving importin-α during nuclear import using a combination of biophysical techniques (biosensor, crystallography, sedimentation equilibrium, electrophoresis, and circular dichroism). Importin-α is shown to exist in a monomeric autoinhibited state (association with NLSs undetectable by biosensor). Association with importin-β (stoichiometry, 1:1; K D = 1.1 × 10 -8 M) increases the affinity for NLSs; the importin-α/β complex binds representative monopartite NLS (simian virus 40 large T-antigen) and bipartite NLS (nucleoplasmin) with affinities (K D = 3.5 × 10 -8 M and 4.8 × 10 -8 M, respectively) comparable with those of a truncated importin-α lacking the autoinhibitory domain (T-antigen NLS, K D = 1.7 × 10 -8 M; nucleoplasmin NLS, K D = 1.4 × 10 -8 M). The autoinhibitory domain (as a separate peptide) binds the truncated importin-α, and the crystal structure of the complex resembles the structure of full-length importin-α. Our results support the model of regulation of nuclear import mediated by the intrasteric autoregulatory sequence of importin-α and provide a quantitative description of the binding and regulatory steps during nuclear import.

An hydrochemical answer to a legislative demand involving health risk related to water quality

Effluents and surface waters around an area involved with the inking of tissues at Itatiba municipality, São Paulo State, Brazil, were chemically analyzed with the purpose of evaluating the influence on the water quality of the chemicals released, as well to provide answers to legislative requirements related to the São Paulo State Register 997 published on 31 May 1976.

Spectrophotometric flow-injection determination of sulphite in white wines involving gas diffusion through a concentric tubular membrane

A flow-injection system is proposed for the spectrophotometric determination of sulphite in white wines. The method involves analyte conversion to SO2, gas diffusion through a Teflon® semi-permeable membrane, collection into an alkaline stream (pH 8), reaction with Malachite green (MG) and monitoring at 620 nm. With a concentric tubular membrane, the system design was simplified. Influence of reagent concentrations, pH of donor and acceptor streams, temperature, timing, surfactant addition and presence of potential interfering species of the wine matrix were investigated. A pronounced (ca. 100%) enhancement in sensitivity was noted by adding cetylpyridinium chloride (CPC). The proposed system is robust and baseline drift is not observed during 4 h operating periods. Only 400 μL of sample and 0.32 mg MG are required per determination. The system handles 30 samples per hour, yielding precise results (r.s.d. < 0.015 for 1.0 - 20.0 mg L-1 SO2) in agreement with those obtained by an alternative procedure.

Thermally developing forced convection of non-Newtonian fluids inside elliptical ducts

Laminar-forced convection inside tubes of various cross-section shapes is of interest in the design of a low Reynolds number heat exchanger apparatus. Heat transfer to thermally developing, hydrodynamically developed forced convection inside tubes of simple geometries such as a circular tube, parallel plate, or annular duct has been well studied in the literature and documented in various books, but for elliptical duct there are not much work done. The main assumptions used in this work are a non-Newtonian fluid, laminar flow, constant physical properties, and negligible axial heat diffusion (high Peclet number). Most of the previous research in elliptical ducts deal mainly with aspects of fully developed laminar flow forced convection, such as velocity profile, maximum velocity, pressure drop, and heat transfer quantities. In this work, we examine heat transfer in a hydrodynamically developed, thermally developing laminar forced convection flow of fluid inside an elliptical tube under a second kind of a boundary condition. To solve the thermally developing problem, we use the generalized integral transform technique (GITT), also known as Sturm-Liouville transform. Actually, such an integral transform is a generalization of the finite Fourier transform, where the sine and cosine functions are replaced by more general sets of orthogonal functions. The axes are algebraically transformed from the Cartesian coordinate system to the elliptical coordinate system in order to avoid the irregular shape of the elliptical duct wall. The GITT is then applied to transform and solve the problem and to obtain the once unknown temperature field. Afterward, it is possible to compute and present the quantities of practical interest, such as the bulk fluid temperature, the local Nusselt number, and the average Nusselt number for various cross-section aspect ratios.

Energy transfer and frequency upconversion involving triads of Pr3+ ions in (Pr3+, Gd3+) doped fluoroindate glass

Blue and ultraviolet luminescence in (Pr3+, Gd3+) doped fluoroindate glass is studied for excitation in the red region (≈590 nm). Frequency upconversion (UC) is observed due to energy transfer (ET) among three Pr3+ ions initially excited to the D21 state corresponding to the ET process D21 + D21 + D21 → S01 + H53 + H53. Additionally, UC luminescence from states P 72 6 and I 72 6 of Gd3+ is observed for an excitation wavelength resonant with transitions of the Pr3+ ions. The characterization of the luminescence signals allowed to determine ET rate among the Pr3+ ions and provides evidence of interconfigurational ET between Gd3+ and Pr3+ ions. © 2006 American Institute of Physics.

Occurrence of Actinobacillus actinomycetemcomitans in patients with chronic periodontitis, aggressive periodontitis, healthy subjects and children with gingivitis in two cities of the state of São Paulo, Brazil

The aim of this study was to determine the frequency of isolation of Actinobacillus actinomycetemcomitans (Aa) in 100 patients with chronic periodontitis, 14 patients with aggressive periodontitis, 142 pre-school children with gingivitis and 134 periodontally healthy subjects. Samples of subgingival plaque were taken using sterilized paper points introduced into periodontal pockets or gingival crevice for 60 seconds and inoculated on TSBV agar, which was incubated under anaerobiosis at 37°C, for 4 days. Microbial identification was performed through biochemical methods and morphocellular and morphocolonial analysis. Aa was detected in 40.3% of healthy subjects, 68% of patients with chronic periodontitis, 92.86% of patients with aggressive periodontitis and 40.14% of children with gingivitis. The rate of recovery of Aa in the tested human groups proved to be higher than previously reported and in agreement with participation of this facultative anaerobe as a member of native microbiota of the periodontium and its relation with aggressive and chronic periodontitis in Brazil.

Extravasation mucocele involving the ventral surface of the tongue (glands of Blandin-Nuhn)

Background. Mucocele is a lesion that involves the salivary glands and respective current ducts caused mainly by traumas in the affected area. Two different histological forms can be found: extravasation phenomenon and mucus-retention cyst where the former is the most frequently observed involving minor salivary glands such as the glands present in the anterior portion of the ventral surface of the tongue (glands of Blandin-Nuhn). Case Report. This report describes a large lesion involving the ventral surface of the tongue that was definitively diagnosed by histological examination as extravasation mucocele. Conclusion. Important concepts are reviewed to help clinicians correctly diagnose and treat this pathology. © 2006 The Authors.

Propriedades mecânicas e de corrosão de dois aços inoxidáveis austeníticos utilizados na fabricação de implantes ortopédicos

ASTM F 138 austenitic stainless steel is extensively used as an orthopedic implant material. However, some aspects, such as low strength in the annealed condition and susceptibility to localized corrosion, limit wider use of this kind of steel. Recently, a high-nitrogen austenitic stainless steel, specified in the standard ISO 5832-9, has been indicated as an alternative to ASTM F 138 steel for more severe loading and permanent application inside the human body. In this work, microstructure, mechanical properties, corrosion resistance and fatigue behavior of both steels were determined and compared. ISO 5832-9 steel displayed better mechanical and corrosion behaviors than did ASTM F 138 steel The combination of these features lead ISO steel to enhanced fatigue performance in both neutral and aggressive environments. Analyzed were the role of nitrogen in solid solution, combined with niobium in the Z-phase, and the factors that led to superior ISO 5832-9 properties.

Biological aspects of Amblyomma brasiliense (Acari: Ixodidae) under laboratory conditions

Although Amblyomma brasiliense Aragão 1908 has been reported as one of the most aggressive ticks to humans in Brazil, information about the biology of this tick species is virtually inexistent. This work reports data on the life cycle of A. brasiliense fed on rabbits and pigs and maintained in an incubator at 20°C, 90% RH and 12 h of light for off-host development. Tick yield of adult females fed on pigs and rabbits was 81.2% and 58.3%, respectively. Females fed on pigs had mean engorgement weight of 862.3 mg and egg mass of 208 mg, while females fed on rabbits had mean engorgement weight of 606.1 mg and egg mass of 160 mg; these values did not differ statistically between host species. Feeding period of female ticks fed on pigs (10 days) was significantly shorter than that on rabbits (17 days). Mean preoviposition period was slightly longer (35.9 days) for ticks fed on pigs than on rabbits (30 days). The minimum incubation period of eggs of ticks from both host species was similar and over 100 days. Egg production efficiency was low for females fed on both hosts (less than 30% and 20% for ticks from pigs and rabbits, respectively). More than 55% of larvae and 79% of nymphs fed on rabbits, set free inside the feeding chambers, engorged successfully. These ticks attained an engorgement weight of 1.3 and 18.2 mg, respectively, and fed for approximately 5 days. The minimum pre-molt period was 30 days for engorged larvae and over 44 days for nymphs. Molting success was low, less than 50% in the case of larvae and less than 20% for nymphs. Further studies are required to better determine the off-host requirements of this tick species. © 2008 Springer Science+Business Media B.V.

Committees for ethics in research involving human subjects

In Brazil since October 1996 there have been guidelines for research involving human subjects. Now human subjects know when their treatment is part of research. Deceit is no longer tolerated. But is not enough to say we offer an explanation to the potential subject and we offer a choice before he or she is confronted with an informed consent form. As in all professional activity, scientific investigation needs social controls. In Brazil, the ultimate responsibility of an investigation lies on the investigator, but in every institution where research is carried out there is a Committee for Ethics in Research. All Committees are subordinated to the National Commission of Ethics in Research, which is submitted to the Brazilian Institute of Health. During 2005 around 17,000 protocols involving 700,000 human subjects were revised by 475 Committees distributed all over the country. Approximately 7,000 people are now working in these Committees.

Sinalização visual e biologia reprodutiva de Dendropsophus werneri (Anura: Hylidae) em área de Mata Atlântica no estado do Paraná, Brasil

We studied the reproductive biology and visual signaling of Dendropsophus werneri, whose distribution is limited to the Atlantic Rain Forest. The fieldwork was carried out in the Estação 2 do IAPAR, municipality of Morretes, state of Paraná, Brazil, from August 2006 to March 2007. Additional information on reproduction was gathered in the Reserva Natural Salto Morato, municipality of Guaraqueçaba, state of Paraná, Brazil, from September 2006 to March 2007. Males were smaller than females. Males called throughout all the study period in Morretes, but were active only during three months in Guaraqueçaba; males called from low vegetation along the edge of temporary ponds in open areas. The visual signaling was observed in two contexts: (1) aggressive behaviors between two males and (2) during the reproduction, by amplectant males. In the territorial behavior, males exhibited both aggressive and mixed calls, visual signaling as well as physical combats. We also recorded satellite behavior in four males. The mean egg number per clutch was 244 ± 32 eggs, varying between 188 and 310 eggs. We observed two reproductive modes: Mode 1 and Mode 24. In the present study, Dendropsophus werneri showed elaborated social interactions involving visual signaling, territorial behavior, mating bahavior with tactile stimuli, and two different reproductive modes, demonstrating its complex reproductive biology.

Dental trauma involving root fracture and periodontal ligament injury: A 10-year retrospective study

The purpose of this retrospective study was to analyze the cases of traumatic dental injuries involving root fracture and/or periodontal ligament injury (except avulsion) treated at the Discipline of Integrated Clinic, School of Dentistry of Araçatuba, São Paulo State University (UNESP), Brazil, from January 1992 to December 2002. Clinical and radiographic records from 161 patients with 287 traumatized teeth that had sustained root fracture and/or injuries to the periodontal ligament were examined. The results of this survey revealed that subluxation (25.09%) was the most common type of periodontal ligament injury, followed by extrusive luxation (19.86%). There was a predominance of young male patients and most of them did not present systemic alterations. Among the etiologic factors, the most frequent causes were falls and bicycle accidents. Injuries on extraoral soft tissues were mostly laceration and abrasion, while gingival and lip mucosa lacerations prevailed on intraoral soft tissues injuries. Radiographically, the most common finding was an increase of the periodontal ligament space. The most commonly performed treatment was root canal therapy. Within the limits of this study, it can be concluded that traumatic dental injuries occur more frequently in young male individuals, due to falls and bicycle accidents. Subluxation was the most common type of periodontal ligament injury. Root canal therapy was the type of treatment most commonly planned and performed.

Effects of the density, aviary type, breed, and age on the behaviors of laying hens kept in cages

An experiment was carried out in order to investigate the behaviors of laying hens due to the environmental factors of: density inside of the cage, aviary type, breed, and age. The experiment was configured as a factorial 4x2x2x2 study, with treatments being four different ages, two different breeds, two different cage densities, and two different aviaries. The birds' behaviors were recorded using video cameras installed in the cages, using samples of 15 minutes recorded from 12 PM to 4 PM. The observed behaviors, frequency and duration of behaviors (measured in seconds) were identified and noted related to each bird. The study was initiated in March 2007, during four non-consecutive weeks. The observed behaviors were: opening wings, stretching, threatening, ruffling feathers, drinking water, aggressive pecking, eating, running, lying down, stretching head out of the cage, preening, mounting, prostrating, and doing nothing (inactivity). Due to the non-normality of the data recorded, the Kruskal-Wallis statistical test of the MINITAB Statistical Software® was used to compare the medians of the variables. For breed factor, only the durations of the eating presented significant differences (p-value< 0.05). For cage density, there was a significant median difference (p-value< 0.05) for almost all behaviors observed. The average length of time of behaviors was higher for the lowest cage density. However, the frequency of behaviors was lmerfor the lowest cage density. The frequency of the behaviors to preen feathers, to lie down, to drink water and to stretch the head were higher in the aviary, where the groups of birds were smaller. The observed behaviors were particularly affected by experimental factors cage density, and aviary type, which directly affects the available space for each bird.

Destroying horseshoes via heterodimensional cycles: Generating bifurcations inside homoclinic classes

In this paper, we propose a model for the destruction of three-dimensional horseshoes via heterodimensional cycles. This model yields some new dynamical features. Among other things, it provides examples of homoclinic classes properly contained in other classes and it is a model of a new sort of heteroclinic bifurcations we call generating. © 2008 Cambridge University Press.