Synthesis of Phthalocyanines-ALA Conjugates: Water-Soluble Compounds with Low Aggregation

Syntheses of two water-soluble phthalocyanines (Pc) containing 5-aminolevulinic acid (ALA) linked to the core structure are described. These compounds were prepared by using original functionalizations, and they present remarkable structural and photophysical features, indicating that they could be applied to photodynamic therapy (PDT).

One-step reverse transcriptase polymerase chain reaction for the diagnosis of respiratory syncytial virus in children

Human respiratory syncytial virus (HRSV) is the main cause of acute lower respiratory tract infections in infants and children. Rapid diagnosis is required to permit appropriate care and treatment and to avoid unnecessary antibiotic use. Reverse transcriptase (RT-PCR) and indirect immunofluorescence assay (IFA) methods have been considered important tools for virus detection due to their high sensitivity and specificity. In order to maximize use-simplicity and minimize the risk of sample cross-contamination inherent in two-step techniques, a RT-PCR method using only a single tube to detect HRSV in clinical samples was developed. Nasopharyngeal aspirates from 226 patients with acute respiratory illness, ranging from infants to 5 years old, were collected at the University Hospital of the University of Sao Paulo (HU-USP), and tested using IFA, one-step RT-PCR, and semi-nested RT-PCR. One hundred and two (45.1%) samples were positive by at least one of the three methods, and 75 (33.2%) were positive by all methods: 92 (40.7%) were positive by one-step RT-PCR, 84 (37.2%) by IFA, and 96 (42.5%) by the semi-nested RT-PCR technique. One-step RT-PCR was shown to be fast, sensitive, and specific for RSV diagnosis, without the added inconvenience and risk of false positive results associated with semi-nested PCR. The combined use of these two methods enhances HRSV detection. (C) 2007 Elsevier B.V. All rights reserved.

Fluxes of CO(2) above a plantation of Eucalyptus in southeast Brazil

Eddy-covariance measurements of net ecosystem exchange of CO(2) (NEE) and estimates of gross ecosystem productivity (GEP) and ecosystem respiration (R(E)) were obtained in a 2-4 year old Eucalyptus plantation during two years with very different winter rainfall In the first (drier) year the annual NEE GEP and RE were lower than the sums in the second (normal) year and conversely the total respiratory costs of assimilated carbon were higher in the dry year than in the normal year Although the net primary production (NPP) in the first year was 23% lower than that of the second year the decrease in the carbon use efficiency (CUE = NPP/GEP) was 11% and autotrophic respiration utilized more resources in the first dry year than in the second normal year The time variations in NEE were followed by NPP because in these young Eucalyptus plantations NEE is very largely dominated by NPP and heterotrophic respiration plays only a relatively minor role During the dry season a pronounced hysteresis was observed in the relationship between NEE and photosynthetically active radiation and NEE fluxes were inversely proportional to humidity saturation deficit values greater than 0 8 kPa Nighttime fluxes of CO(2) during calm conditions when the friction velocity (u) was below the threshold (0 25 ms(-1)) were estimated based on a Q(10) temperature-dependence relationship adjusted separately for different classes of soil moisture content which regulated the temperature sensitivity of ecosystem respiration (C) 2010 Elsevier B V All rights reserved

The energy and water balance of a Eucalyptus plantation in southeast Brazil

The eddy covariance method was used to measure energy and water balance of a plantation of Eucalyptus (grandis x urophylla) hybrids over a 2 year period. The average daily evaporation rates were 5.4 (+/- 2.0) mm day(-1) in summer, but fell to 1.2 (+/- 0.3) mm day(-1) in winter. In contrast, the sensible heat flux was relatively low in summer but dominated the energy balance in winter. Evaporation accounted for 80% and 26% of the available energy, in summer and winter respectively. The annual evaporation was 82% (1124 mm) and 96% (1235 mm) of the annual rainfall recorded during the first and second year, respectively. Daily average canopy and aerodynamic conductance to water vapour were in the summer 51.9 (+/- 38.4) mm s(-1) 84.1 (+/- 25.6) mm s(-1), respectively; and in the winter 6.0 (+/- 10.5) mm s(-1) and 111.6 (+/- 24.6) mm s(-1), respectively. (C) 2010 Elsevier B.V. All rights reserved.

Downward longwave radiation estimates for clear and all-sky conditions in the Sertozinho region of So Paulo, Brazil

The accurate estimate of the surface longwave fluxes contribution is important for the calculation of the surface radiation budget, which in turn controls all the components of the surface energy budget, such as evaporation and the sensible heat fluxes. This study evaluates the performance of the various downward longwave radiation parameterizations for clear and all-sky days applied to the Sertozinho region in So Paulo, Brazil. Equations have been adjusted to the observations of longwave radiation. The adjusted equations were evaluated for every hour throughout the day and the results showed good fits for most of the day, except near dawn and sunset, followed by nighttime. The seasonal variation was studied by comparing the dry period against the rainy period in the dataset. The least square linear regressions resulted in coefficients equal to the coefficients found for the complete period, both in the dry period and in the rainy period. It is expected that the best fit equation to the observed data for this site be used to produce estimates in other regions of the State of So Paulo, where such information is not available.

Cladistic analysis of the Stygninae and description of a new species of Protimesius Roewer, 1913 (Opiliones: Stygnidae)

A new species of Stygnidae is described from the state of Bahia, Brazil. Protimesius bahiensis sp. nov. can be distinguished from the remaining species of the genus by the combination of: male femur IV unarmed and cylindrical; male patella IV with a row of large dorsal acute tubercles, increasing in size distally and male tibia IV with one mesodistal tubercle; ventral plate of the penis with three pairs of distal curved setae and one pair of intermediate setae, smaller than the rest. A cladistic analysis of the subfamily is presented. Stygninae is divided in two groups of genera: (Ricstygnus, Stygnus, Sickesia), with a wide distribution and (Pickeliana (Protimesius (Phareus (Stenophareus (Auranus (Verrucastygnus, Stenostygnoides)))))), associated to the Guiana Shield, Amazon basin and Northeastern Brazil. The monophyly of Protimesius is supported by the apex of pedipalpal tibia sockets bifid (homoplastically present in Verrucastygnus and Stenostygnoides) and by the presence of scopulae with non-spatulated hairs.

A new species of Cycloramphus Tschudi (Anura: Cycloramphidae) from the Parque Nacional da Serra dos Orgaos, Southeastern Brazil

We report here the discovery of a new species of frog associated to the open areas of the highlands of the Parque Nacional da Serra dos Orgaos. The new species, Cycloramphus organensis is characterized by a unique skin texture, medium size ( maximum male and female SVL 26.4 mm and 33.3 mm respectively), dorsal surfaces uniformly brick red colored, uniformly areolate skin on dorsum, pupil horizontal, iris with a menisc on upper margin; no fleshy tubercles on eyelid, tympanic annulus concealed beneath skin, macroglands not visible externally, fingers and toes without fringes and webs; supernumerary palmar and plantar tubercles absent, nuptial spines absent. Despite the presence of an iris menisc, a character shared by frogs of both genera Cycloramphus and Zachaenus Cope, the combination of morphological characters is so unique that the allocation of the species to any of these genera remains ambiguous. Consequently, we used additional molecular-based phylogenetic analyses to ascertain the position of the new taxon. The new species proved to be embedded within the genus Cycloramphus.

The non-coding RNA BC1 is down-regulated in the hippocampus of Wistar Audiogenic Rat (WAR) strain after audiogenic kindling

The aim of this study was to identify molecular pathways involved in audiogenic seizures in the epilepsy-prone Wistar Audiogenic Rat (WAR). For this, we used a suppression-subtractive hybridization (SSH) library from the hippocampus of WARs coupled to microarray comparative gene expression analysis, followed by Northern blot validation of individual genes. We discovered that the levels of the non-protein coding (npc) RNA BC1 were significantly reduced in the hippocampus of WARs submitted to repeated audiogenic seizures (audiogenic kindling) when compared to Wistar resistant rats and to both naive WARs and Wistars. By quantitative in situ hybridization, we verified lower levels of BC1 RNA in the GD-hilus and significant signal ratio reduction in the stratum radiatum and stratum pyramidale of hippocampal CA3 subfield of audiogenic kindled animals. Functional results recently obtained in a BC1-/- mouse model and our current data are supportive of a potential disruption in signaling pathways, upstream of BC1, associated with the seizure susceptibility of WARs. (C) 2010 Elsevier B.V. All rights reserved.

Increased Expression of GluR2-flip in the Hippocampus of the Wistar Audiogenic Rat Strain After Acute and Kindled Seizures

The Wistar Audiogenic Rat (WAR) is an epileptic-prone strain developed by genetic selection from a Wistar progenitor based on the pattern of behavioral response to sound stimulation. Chronic acoustic stimulation protocols of WARs (audiogenic kindling) generate limbic epileptogenesis, confirmed by ictal semiology, amygdale, and hippocampal EEG, accompanied by hippocampal and amygdala cell loss, as well as neurogenesis in the dentate gyrus (DG). In an effort to identify genes involved in molecular mechanisms underlying epileptic process, we used suppression-subtractive hybridization to construct normalized cDNA library enriched for transcripts expressed in the hippocampus of WARs. The most represented gene among the 133 clones sequenced was the ionotropic glutamate receptor subunit II (GluR2), a member of the a-amino-3-hydroxy-5-methyl-4-isoxazoleopropionic acid (AMPA) receptor. Although semiquantitative RT-PCR analysis shows that the hippocampal levels of the GluR2 subunits do not differ between naive WARs and their Wistar counterparts, we observed that the expression of the transcript encoding the splice-variant GluR2-flip is increased in the hippocampus of WARs submitted to both acute and kindled audiogenic seizures. Moreover, using in situ hybridization, we verified upregulation of GluR2-flip mainly in the CA1 region, among the hippocampal subfields of audiogenic kindled WARs. Our findings on differential upregulation of GluR2-flip isoform in the hippocampus of WARs displaying audiogenic seizures is original and agree with and extend previous immunohistochemical for GluR2 data obtained in the Chinese P77PMC audiogenic rat strain, reinforcing the association of limbic AMPA alterations with epileptic seizures. (C) 2009 Wiley-Liss, Inc.

Glucose metabolism by lymphocytes, macrophages, and tumor cells from Walker 256 tumor-bearing rats supplemented with fish oil for one generation

Here we investigated the effect of lifelong supplementation of the diet with coconut fat (CO, rich in saturated fatty acids) or fish oil (170, rich in n-3 polyunsaturated fatty acids) on tumor growth and lactate production from glucose in Walker 256 tumor cells, peritoneal macrophages, spleen, and gut-associated lymphocytes. Female Wistar rats were supplemented with CO or FO prior to mating and then throughout pregnancy and gestation and then the male offspring were supplemented from weaning until 90 days of age. Then they were inoculated subcutaneously with Walker 256 tumor cells. Tumor weight at 14 days in control rats (those fed standard chow) and CO supplemented was approximately 30 g. Supplementation of the diet with FO significantly reduced tumor growth by 76%. Lactate production (nmol h(-1) mg(-1) protein) from glucose by Walker 256 cells in the group fed regular chow (W) was 381.8 +/- 14.9. Supplementation with coconut fat (WCO) caused a significant reduction in lactate production by 1.6-fold and with fish oil (WFO) by 3.8-fold. Spleen lymphocytes obtained from W and WCO groups had markedly increased lactate production (553 +/- 70 and 635 +/- 150) when compared to non-tumor-bearing rats (similar to 260 +/- 30). FO supplementation reduced significantly the lactate production (297 +/- 50). Gut-associated lymphocytes obtained from W and WCO groups increased lactate production markedly (280 +/- 31 and 276 +/- 25) when compared to non-tumor-bearing rats (similar to 90 +/- 18). FO supplementation reduced significantly the lactate production (168 +/- 14). Lactate production by peritoneal macrophages was increased by tumor burden but there was no difference between the groups fed the various diets. Lifelong consumption of FO protects against tumor growth and modifies glucose metabolism in Walker tumor cells and lymphocytes but not in macrophages. Copyright (C) 2008 John Wiley & Sons, Ltd.

Unusual Presentation of Brain Aspergillosis in Chronic Granulomatous Disease

Aspergillus is a frequently observed pathogen in patients with chronic granulomatous disease. We report on a patient with chronic granulomatous disease and severe brain aspergillosis with an unusual presentation and favorable course. We discuss the impact of this infection on morbidity and mortality, adequate therapeutic management, and the need to investigate a possible fungal infection, despite nonspecific signs. (C) 2010 by Elsevier Inc. All rights reserved.

Autolytic Mycobacterium leprae Hsp65 fragments may act as biological markers for autoimmune diseases

Investigating the proteolytic activity of the recombinant Mycobacterium leprae Heat Shock Protein of 65 kDa (rHsp65), chaperonin 2 (cpn2), we observed that it displays high instability. The fragmentation process starts at the C-terminus followed by progressive degradation of the N-terminus, which leads to a stable fragment comprising the middle region of the molecule. Urea was able to prevent autolysis, probably due to its denaturing action, while EDTA increased degradation levels indicating the need for metal ions. Peptides originated from autolysis were purified and analyzed by mass spectrometry, generating a continuous map. Since the bacteria and mammalian Hsp60 are known to be targets of the immune response and have been implicated in autoimmune diseases and chronic inflammation, the in vivo effect of rHsp65 peptides was evaluated in the spontaneous Systemic Lupus Erythematosus (SLE) model developed by the (NZB/NZW)F(1) mouse hybrids, and their individual anti-rHsp65 IgG2a/IgG1 antibody titer ratio was determined. The results showed orientation toward a T(H)1 responsiveness, and the treatment with the rHsp65 peptides diminished the environmental variance of the survival time of treated animals. These results outline the fact that environmental factors may also act through the modified stability expression of Heat Shock Proteins intervening during autoimmune processes. (C) 2011 Elsevier Ltd. All rights reserved.

Strategies to Optimize Biosensors Based on Impedance Spectroscopy to Detect Phytic Acid Using Layer-by-Layer Films

Impedance spectroscopy has been proven a powerful tool for reaching high sensitivity in sensor arrays made with nanostructured films in the so-called electronic tongue systems, whose distinguishing ability may be enhanced with sensing units capable of molecular recognition. In this study we show that for optimized sensors and bio-sensors the dielectric relaxation processes involved in impedance measurements should also be considered, in addition to an adequate choice of sensing materials. We used sensing units made from layer-by-layer (LbL) films with alternating layers of the polyeletrolytes, poly(allylamine) hydrochloride (PAH) and poly(vinyl sulfonate) (PVS), or LbL films of PAH alternated with layers of the enzyme phytase, all adsorbed on gold interdigitate electrodes. Surprisingly, the detection of phytic acid was as effective in the PVS/PAH sensing system as with the PAH/phytase system, in spite of the specific interactions of the latter. This was attributed to the dependence of the relaxation processes on nonspecific interactions such as electrostatic cross-linking and possibly on the distinct film architecture as the phytase layers were found to grow as columns on the LbL film, in contrast to the molecularly thin PAH/PVS films. Using projection techniques, we were able to detect phytic acid at the micromolar level with either of the sensing units in a data analysis procedure that allows for further optimization.

Use of Information Visualization Methods Eliminating Cross Talk in Multiple Sensing Units Investigated for a Light-Addressable Potentiometric Sensor

The integration of nanostructured films containing biomolecules and silicon-based technologies is a promising direction for reaching miniaturized biosensors that exhibit high sensitivity and selectivity. A challenge, however, is to avoid cross talk among sensing units in an array with multiple sensors located on a small area. In this letter, we describe an array of 16 sensing units, of a light-addressable potentiometric sensor (LAPS), which was made with layer-by-Layer (LbL) films of a poly(amidomine) dendrimer (PAMAM) and single-walled carbon nanotubes (SWNTs), coated with a layer of the enzyme penicillinase. A visual inspection of the data from constant-current measurements with liquid samples containing distinct concentrations of penicillin, glucose, or a buffer indicated a possible cross talk between units that contained penicillinase and those that did not. With the use of multidimensional data projection techniques, normally employed in information Visualization methods, we managed to distinguish the results from the modified LAPS, even in cases where the units were adjacent to each other. Furthermore, the plots generated with the interactive document map (IDMAP) projection technique enabled the distinction of the different concentrations of penicillin, from 5 mmol L(-1) down to 0.5 mmol L(-1). Data visualization also confirmed the enhanced performance of the sensing units containing carbon nanotubes, consistent with the analysis of results for LAPS sensors. The use of visual analytics, as with projection methods, may be essential to handle a large amount of data generated in multiple sensor arrays to achieve high performance in miniaturized systems.

Poly-bagging predictors for classification modelling for credit scoring

Credit scoring modelling comprises one of the leading formal tools for supporting the granting of credit. Its core objective consists of the generation of a score by means of which potential clients can be listed in the order of the probability of default. A critical factor is whether a credit scoring model is accurate enough in order to provide correct classification of the client as a good or bad payer. In this context the concept of bootstraping aggregating (bagging) arises. The basic idea is to generate multiple classifiers by obtaining the predicted values from the fitted models to several replicated datasets and then combining them into a single predictive classification in order to improve the classification accuracy. In this paper we propose a new bagging-type variant procedure, which we call poly-bagging, consisting of combining predictors over a succession of resamplings. The study is derived by credit scoring modelling. The proposed poly-bagging procedure was applied to some different artificial datasets and to a real granting of credit dataset up to three successions of resamplings. We observed better classification accuracy for the two-bagged and the three-bagged models for all considered setups. These results lead to a strong indication that the poly-bagging approach may promote improvement on the modelling performance measures, while keeping a flexible and straightforward bagging-type structure easy to implement. (C) 2011 Elsevier Ltd. All rights reserved.