DOES THE CHRONOTYPE CLASSIFICATION NEED TO BE UPDATED? PRELIMINARY FINDINGS

Traditionally, chronotype classification is based on the Morningness-Eveningness Questionnaire (MEQ). It is implicit in the classification that intermediate individuals get intermediate scores to most of the MEQ questions. However, a small group of individuals has a different pattern of answers. In some questions, they answer as ""morning-types"" and in some others they answer as ""evening-types,"" resulting in an intermediate total score. ""Evening-type"" and ""Morning-type"" answers were set as A(1) and A(4), respectively. Intermediate answers were set as A(2) and A(3). The following algorithm was applied: Bimodality Index = (Sigma A(1) x Sigma A(4))(2) - (Sigma A(2) x Sigma A(3))(2). Neither-types that had positive bimodality scores were classified as bimodal. If our hypothesis is validated by objective data, an update of chronotype classification will be required. (Author correspondence: brunojm@ymail.com)

Electrical Stimulation During Gait Promotes Increase of Muscle Cross-sectional Area in Quadriplegics: A Preliminary Study

Increases in muscular cross-sectional area (CSA) occur in quadriplegics after training, but the effects of neuromuscular electrical stimulation (NMES) along with training are unknown. Thus, we addressed two questions: (1) Does NMES during treadmill gait training increase the quadriceps CSA in complete quadriplegics?; and (2) Is treadmill gait training alone enough to observe an increase in CSA? Fifteen quadriplegics were divided into gait (n = 8) and control (n = 7) groups. The gait group performed training with NMES for 6 months twice a week for 20 minutes each time. After 6 months of traditional therapy, the control group received the same gait training protocol but without NMES for an additional 6 months. Axial images of the thigh were acquired at the beginning of the study, at 6 months (for both groups), and at 12 months for the control group to determine the average quadriceps CSA. After 6 months, there was an increase of CSA in the gait group (from 49.8 +/- A 9.4 cm(2) to 57.3 +/- A 10.3 cm(2)), but not in the control group (from 43.6 +/- A 7.6 cm(2) to 41.8 +/- A 8.4 cm(2)). After another 6 months of gait without NMES in the control group, the CSA did not change (from 41.8 +/- A 8.4 cm(2) to 41.7 +/- A 7.9 cm(2)). The increase in quadriceps CSA after gait training in patients with chronic complete quadriplegia appears associated with NMES.

Inflammation markers in healthy and periodontitis patients: a preliminary data screening

Advances in diagnostic research are moving towards methods whereby the periodontal risk can be identified and quantified by objective measures using biomarkers. Patients with periodontitis may have elevated circulating levels of specific inflammatory markers that can be correlated to the severity of the disease. The purpose of this study was to evaluate whether differences in the serum levels of inflammatory biomarkers are differentially expressed in healthy and periodontitis patients. Twenty-five patients (8 healthy patients and 17 chronic periodontitis patients) were enrolled in the study. A 15 mL blood sample was used for identification of the inflammatory markers, with a human inflammatory flow cytometry multiplex assay. Among 24 assessed cytokines, only 3 (RANTES, MIG and Eotaxin) were statistically different between groups (p<0.05). In conclusion, some of the selected markers of inflammation are differentially expressed in healthy and periodontitis patients. Cytokine profile analysis may be further explored to distinguish the periodontitis patients from the ones free of disease and also to be used as a measure of risk. The present data, however, are limited and larger sample size studies are required to validate the findings of the specific biomarkers.

Successful transplant of mesenchymal stem cells in induced osteonecrosis of the ovine femoral head: preliminary results

PURPOSE: Evaluate the bone tissue recovery following transplantation of ovine mesenchymal stem cells (MSC) from bone marrow and human immature dental-pulp stem cells (hIDPSC) in ovine model of induced osteonecrosis of femoral head (ONFH). METHODS: Eight sheep were divided in three experimental groups. First group was composed by four animals with ONFH induced by ethanol through central decompression (CD), for control group without any treatment. The second and third group were compose by two animals, six weeks after ONFH induction received transplantation of heterologous ovine MSC (CD + oMSC), and hIDPSC (CD + hIDPSC), respectively. In both experiments the cells were transplanted without application of any type of immunosupression protocol. RESULTS: Our data indicate that both cell types used in experiments were able to proliferate within injured site providing bone tissue recovery. The histological results obtained from CD+hIDPSC suggested that the bone regeneration in such animals was better than that observed in CD animals. CONCLUSION: Mesenchymal stem cell transplant in induced ovine osteonecrosis of femoral head by central decompression technique is safe, and apparently favors bone regeneration of damaged tissues.

Alveolar osseous defect in rat for cell therapy: preliminary report

PURPOSE: To study were to reproduce an alveolar bone defect model in Wistar rats to be used for testing the efficacy of stem cell therapies. Additionally, we also aimed to determine the osteogenesis process of this osseous defect in the 1 month period post-surgery. METHODS: The animals were randomly divided into two groups of 7 animals each. A gingivobuccal incision was made, and a bone defect of 28 mm² of area was performed in the alveolar region. Animals were killed at 2 weeks after surgery (n=7) and 4 weeks after surgery (n=7). RESULTS: The average area of the alveolar defect at time point of 2 weeks was 22.27 ± 1.31 mm² and the average area of alveolar defect at time point of 4 weeks was 9.03 ± 1.17 mm². The average amount of bone formation at time point of 2 weeks was 5.73 ± 1.31 mm² and the average amount of bone formation at time point of 4 weeks was 19 ± 1.17 mm². Statistically significant differences between the amount of bone formation at 2 weeks and 4 weeks after surgery were seen (p=0.003). CONCLUSION: The highest rate of ossification occurred mostly from 2 to 4 weeks after surgery. This observation suggests that 4 weeks after the bone defect creation should be a satisfactory timing to assess the potential of bone inductive stem cells to accelerate bone regeneration in Wistar rats.

A Classificação Internacional de Funcionalidade, Incapacidade e Saúde e a AIDS: uma proposta de core set

O advento da terapia anti-retroviral de alta potência (HAART) alterou a história natural da aids, diminuindo sua mortalidade e a incidência de doenças oportunistas e aumentando a esperança de vida das pessoas vivendo com aids.Como uma doença crônica, outras questões passam a ser relevantes, entre elas a adesão ao tratamento, seus efeitos adversos e a qualidade de vida das pessoas nessa condição. A CIF constitui um instrumento adequado para identificar as características da funcionalidade, do ambiente e condições pessoais que interferem na qualidade de vida. Instrumentos para a sua aplicação, core sets, têm sido desenvolvidos para várias condições de saúde. Com o objetivo de propor um core set para aids, foram desenvolvidas duas etapas preliminares do modelo proposto para a construção desses instrumentos. A primeira etapa, de revisão sistemática buscou no MEDLINE artigos com descritores HAART e qualidade de vida, publicados em inglês, de 2000 a 2004. Foram selecionados 31 estudos que resultou em 87 conceitos dos quais 66 puderam ser identificados como categorias da CIF. Estas formaram as perguntas da entrevista aplicada em 42 voluntários, pacientes de um centro de referência para DST e Aids de São Paulo. Entre as condições mais freqüentemente associadas ao tratamento, estão às mudanças na imagem corporal, conseqüência da lipodistrofia, apontada em 84 por cento dos estudos e em 93 por cento das entrevistas. Alterações das funções digestivas, das relações íntimas, e das funções sexuais foram condições importantes identificadas no estudo. As duas etapas definiram 40 categorias da CIF como proposta preliminar de um core set para pacientes com aids

Preliminary biocompatibility investigation of magnetic albumin nanosphere designed as a potential versatile drug delivery system

Background: The magnetic albumin nanosphere (MAN), encapsulating maghemite nanoparticles, was designed as a magnetic drug delivery system (MDDS) able to perform a variety of biomedical applications. It is noteworthy that MAN was efficient in treating Ehrlich's tumors by the magnetohyperthermia procedure. Methods and materials: In this study, several nanotoxicity tests were systematically carried out in mice from 30 minutes until 30 days after MAN injection to investigate their biocompatibility status. Cytometry analysis, viability tests, micronucleus assay, and histological analysis were performed. Results: Cytometry analysis and viability tests revealed MAN promotes only slight and temporary alterations in the frequency of both leukocyte populations and viable peritoneal cells, respectively. Micronucleus assay showed absolutely no genotoxicity or cytotoxicity effects and histological analysis showed no alterations or even nanoparticle clusters in several investigated organs but, interestingly, revealed the presence of MAN clusters in the central nervous system (CNS). Conclusion: The results showed that MAN has desirable in vivo biocompatibility, presenting potential for use as a MDDS, especially in CNS disease therapy.

Some questions about the Metropolitan Areas in Buenos Aires, Sao Paulo and Santiago regions

Among the numerous problems that are common to the Latin-American metropolises, such as the deep socio-spatial segregation, the impressive territorial fragmentation and the real estate valorisation that overvalues some territories, whilst it depreciates others, we have chosen to focus on the management of the metropolitan regions. That question clearly indicates that due to the great current changes of the economical restructuring - a process that strengthened the capitalist logic of social development - the traditional form of thinking urban planning has found its limits. Consequently, this issue of metropolitan management shows the need to look for new ways of metropolitan administration that can answer to the The main metropolitan regions of South America: Buenos Aires, Sao Paulo and Santiago form our references to characterize the recent changes from a territorial point of view on the one hand; and relative to the new determinations of the metropolis on the other hand. This leads us to discuss the challenges that metropolitan management face in a scenario of governability fragmentation.

Effect of Nd:YAG Laser and Acidulated Phosphate Fluoride on Bovine and Human Enamel Submitted to Erosion/Abrasion or Erosion Only: An in Vitro Preliminary Study

Objective: The aim of the present in vitro study was to evaluate, using two different methodologies, the effectiveness of pulsed Nd:YAG laser irradiation associated with topical acidulated phosphate fluoride (APF) for preventing enamel erosion and structure loss under regimes of erosion and abrasion or erosion only. Background Data: An increased incidence of noncarious lesions (erosion and abrasion) has been observed, consequently new preventative therapies have been proposed. Materials and Methods: Two different methodologies were performed. For the first, 100 bovine crowns were submitted to four different treatments (n = 25): no treatment (control), 4 min application of APF, Nd:YAG laser irradiation (1 W, 100 mJ, 10 Hz, 141.5 J/cm(2)), and Nd:YAG laser irradiation+4 min of APF. After the specimens were exposed to citric acid (2% w/v; 30 min), they were submitted to 5000 brushing cycles. Specimen mass was measured before and after the treatments. For the second methodology, 20 human crowns were embedded in acrylic resin and cut surfaces were exposed and polished. The specimens were divided into four groups (n = 10): no treatment (control), APF for 4 min, Nd:YAG laser irradiation (1 W, 100 mJ, 10 Hz, 125 J/cm(2)), and Nd:YAG laser irradiation+APF. The samples were then immersed in citric acid (2% w/v; 90 min). Vickers hardness was obtained before and after the treatments. Results: The Nd:YAG laser irradiation+APF (bovine and human enamel) was more effective and yielded statistically significant results for surface microhardness and enamel wear. Conclusion: Nd:YAG laser irradiation associated with APF reduced bovine enamel wear and human enamel softening when samples were submitted to a regime of erosion and abrasion or erosion only in vitro.

Expression, crystallization and preliminary crystallographic analysis of PilZ(XAC1133) from Xanthomonas axonopodis pv. citri

Proteins containing PilZ domains are widespread in Gram-negative bacteria and have recently been shown to be involved in the control of biofilm formation, adherence, aggregation, virulence-factor production and motility. Furthermore, some PilZ domains have recently been shown to bind the second messenger bis(3'-> 5') cyclic diGMP. Here, the cloning, expression, purification and crystallization of PilZ(XAC1133), a protein consisting of a single PilZ domain from Xanthomonas axonopodis pv. citri, is reported. The closest PilZ(XAC1133) homologues in Pseudomonas aeruginosa and Neisseria meningitidis control type IV pilus function. Recombinant PilZ(XAC1133) containing selenomethionine was crystallized in space group P6(1). The unit-cell parameters were a = 62.125, b = 62.125, c = 83.543 angstrom. These crystals diffracted to 1.85 angstrom resolution and a MAD data set was collected at a synchrotron source. The calculated Matthews coefficient suggested the presence of two PilZ(XAC1133) molecules in the asymmetric unit.

Crystallization and preliminary X-ray analysis of LipL32 from Leptospira interrogans serovar Copenhageni

LipL32 is a major surface protein that is expressed during infection by pathogenic Leptospira. Here, the crystallization of recombinant LipL32(21-272), which corresponds to the mature LipL32 protein minus its N-terminal lipid-anchored cysteine residue, is described. Selenomethionine-labelled LipL32(21-272) crystals diffracted to 2.25 angstrom resolution at a synchrotron source. The space group was P3(1)21 or P3(2)21 and the unit-cell parameters were a = b = 126.7, c = 96.0 angstrom.

Crystallization and preliminary X-ray diffraction analysis of a glutathione S-transferase from Xylella fastidiosa

Glutathione S-transferases (GSTs) form a group of multifunctional isoenzymes that catalyze the glutathione-dependent conjugation and reduction reactions involved in the cellular detoxification of xenobiotic and endobiotic compounds. GST from Xylella fastidiosa (xfGST) was overexpressed in Escherichia coli and purified by conventional affinity chromatography. In this study, the crystallization and preliminary X-ray analysis of xfGST is described. The purified protein was crystallized by the vapour-diffusion method, producing crystals that belonged to the triclinic space group P1. The unit-cell parameters were a = 47.73, b = 87.73, c = 90.74 angstrom, alpha = 63.45, beta = 80.66, gamma = 94.55 degrees. xfGST crystals diffracted to 2.23 angstrom resolution on a rotating-anode X-ray source.

Crystallization and preliminary structural analysis of the giant haemoglobin from Glossoscolex paulistus at 3.2 angstrom

Glossoscolex paulistus is a free-living earthworm encountered in south-east Brazil. Its oxygen transport requirements are undertaken by a giant extracellular haemoglobin, or erythrocruorin (HbGp), which has an approximate molecular mass of 3.6 MDa and, by analogy with its homologue from Lumbricus terrestris (HbLt), is believed to be composed of a total of 180 polypeptide chains. In the present work the full 3.6 MDa particle in its cyanomet state was purified and crystallized using sodium citrate or PEG8000 as precipitant. The crystals contain one-quarter of the full particle in the asymmetric unit of the I222 cell and have parameters of a = 270.8 angstrom, b = 320.3 angstrom and c = 332.4 angstrom. Diffraction data were collected to 3.15 angstrom using synchrotron radiation on beamline X29A at the Brookhaven National Laboratory and represent the highest resolution data described to date for similar erythrocruorins. The structure was solved by molecular replacement using a search model corresponding to one-twelfth of its homologue from HbLt. This revealed that HbGp belongs to the type I class of erythrocruorins and provided an interpretable initial electron density map in which many features including the haem groups and disulfide bonds could be identified.

Purification, crystallization and preliminary crystallographic analysis of the catalytic domain of the extracellular cellulase CBHI from Trichoderma harzianum

The filamentous fungus Trichoderma harzianum has a considerable cellulolytic activity that is mediated by a complex of enzymes which are essential for the hydrolysis of microcrystalline cellulose. These enzymes were produced by the induction of T. harzianum with microcrystalline cellulose (Avicel) under submerged fermentation in a bioreactor. The catalytic core domain (CCD) of cellobiohydrolase I (CBHI) was purified from the extracellular extracts and submitted to robotic crystallization. Diffraction-quality CBHI CCD crystals were grown and an X-ray diffraction data set was collected under cryogenic conditions using a synchrotron-radiation source.

Protein preparation, crystallization and preliminary X-ray analysis of Trypanosoma cruzi nucleoside diphosphate kinase 1

The flagellated protozoan parasite Trypanosoma cruzi is the aetiological agent of Chagas disease. Nucleoside diphosphate kinases (NDPKs) are enzymes that are involved in energy management and nucleoside balance in the cell. T. cruzi TcNDPK1, a canonical isoform, was overexpressed in Escherichia coli as an N-terminally poly-His-tagged fusion protein and crystallized. Crystals grew after 72 h in 0.2 M MgCl(2), 20% PEG 3350. Data were collected to 3.5 angstrom resolution using synchrotron X-ray radiation at the National Synchrotron Light Laboratory (Campinas, Brazil). The crystals belonged to the trigonal space group P3, with unit-cell parameters a = b = 127.84, c = 275.49 angstrom. Structure determination is under way and will provide relevant information that may lead to the first step in rational drug design for the treatment of Chagas disease.