Phenotypic and functional evaluations of peripheral blood monocytes from chronic-form paracoccidioidomycosis patients before and after treatment

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

Serological Diagnosis of Paracoccidioidomycosis: High Rate of Inter-laboratorial Variability among Medical Mycology Reference Centers

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

Análise funcional do gene da proteína 14-3-3 de Paracoccidioides brasiliensis

A paracoccidioidomicose (PCM) é micose profunda causada pelo fungo dimórfico Paracoccidioides brasiliensis (Pb), endêmica na América Latina, principalmente no Brasil. A capacidade de P. brasiliensis de não só provocar doença humana, mas também de causar micose com grande variedade de manifestações clínicas, desde formas localizadas até doença disseminada, evoluindo para letalidade, depende provavelmente da virulência do fungo, da habilidade deste em interagir com as estruturas superficiais do hospedeiro e invadi-las, e da resposta imunológica deste último. O sucesso da colonização dos tecidos do hospedeiro pelo fungo é um evento complexo, geralmente envolvendo um ligante codificado pelo patógeno (adesinas) e um receptor da célula. Uma adesina de 30 kDa de P. brasiliensis, ligante de laminina, foi caracterizada através de seqüenciamento de aminoácidos, mostrando que esta é uma proteína 14-3-3 envolvida na adesão deste fungo às células epiteliais. Estas formam uma família de proteínas diméricas, ácidas e estão presentes em múltiplas isoformas em muitos organismos eucariotos. Com o intuito de se estudar sua funcionalidade em P. brasiliensis, pretendeu-se clonar, caracterizar e utilizar como hospedeiro do gene desta proteína a levedura Saccharomyces cerevisiae. Para tanto, as seqüências gênicas da adesina 14-3-3 de isolados de P. brasiliensis obtida pela clonagem do cDNA em vetores bacterianos foram utilizadas para obtenção de um homólogo funcional em S. cerevisiae. A capacidade do gene da 14-3-3 de P. brasiliensis ser um homólogo funcional, aderir e invadir as células epiteliais tratadas deve ser avaliada utilizando o modelo pré-existente de culturas celulares in vitro de linhagens humanas. Assim, neste estudo além da clonagem, expressão da 14-3-3 de Pb e obtenção do homólogo funcional do gene desta proteína em S. cerevisiae, foi iniciada... (Resumo completo, clicar acesso eletrônico abaixo)

Adesão de Paracoccidioides brasiliensis e outros fungos anemófilos a teias de aranhas: implicações para a identificação de nichos microbianos no ambiente

The dimorphic pathogenic fungus, Paracoccidioides brasiliensis (Pb), is the etiological agent of paracoccidioidomycosis (PCM), the most important systemic mycosis in Latin America. While the yeast phase can be isolated from patients affected by paracoccidioidomycosis, dogs and naturally infected armadillos; several elements related to the ecology of the saprophytic phase of the pathogen, which is responsible for the production of infective propagules, are poorly understood, hampering the adoption of preventive measures. The demonstration of the high incidence of Pb infection in the 9- banded armadillo, Dasypus novemcinctus, has opened new perspectives for the identification of the pathogen’s habitat. At the opening of the armadillos’ burrows, spider webs are commonly found. The objective of this study was to evaluate the presence of Pb in spider webs samples related to the habitat of armadillos. Spider web samples were collected at Lageado Farm, Botucatu/SP and prepared for microscopic, molecular and mycological analyses. Microscopic analysis showed that different fungi were closely attached to spider web samples. Nested-PCR reaction showed positive amplification for Pb in 4 samples, with identity confirmed by amplicon sequencing. Fungal colonies also included members of Aspergillus, Blastobotrys, Penicillium, Candida, and Sporothrix genera, which are related to opportunistic disease and primary infections of great medical importance. In vitro adhesion tests of mycelia and yeast form of Pb to the spider webs were also performed, in order to analyze the possible physical attraction between fungal cells and the spider web protein network. The results showed a clear adherence of fungal particles to spider webs. In the current literature, there are no studies reporting adhesive properties of microorganisms to spider webs... (Complete abstract click electronic access below)

Análise funcional da proteína 14-3-3 de Paracoccidioides brasiliensis e prospecção de alvos terapêuticos inibidores da interação de P. brasiliensis à pneumócitos

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

Identificação de Neutrophil Extracellular Traps em lesões tegumentares de pacientes com Paracoccidiodomicose e análise da formação de NETs in vitro por neutrófilos humanos desafiados com Paracoccidioides brasiliensis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Caracterização e identificação de moléculas de superfície presentes em Paracoccidioides spp

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

A method for retrieving endodontic or atypical non-endodontic separated instruments from the root canal: a report of two cases

Aim: This clinical report presents a new method for retrieving separated instruments from the root canal with minimally invasive procedures. Background: The presence of separated instrument in root canal may interfere in the endodontic treatment prognosis. There are several recommended methods to retrieve separated instruments, but some are difficult in clinically practice. Case report: This study describes two cases of separated instrument removal from the root canal using a stainless-steel prepared needle associated with a K-file. Case 1 presented a fractured gutta-percha condenser within the mandibular second premolar, it was separated during incorrect intracanal medication calcium hydroxide placement. Case 2 had a fractured sewing needle within the upper central incisor that the patient used to remove food debris from the root canal. After cervical preparation, the fractured instruments were fitted inside a prepared needle and then an endodontic instrument (#25 K-file) was adapted with clockwise turning motion between the needle inner wall and the fragment. Conclusion: The endodontic or atypical nonendodontic separated instrument may be easily pull on of the root canal using a single and low cost device. Clinical significance: The methods for retrieving separated instruments from root canal are difficult and destructive procedures. The present case describes a simple method to solve this problem.

Ability of different methods to fill retrograde cavities with MTA.

The Mineral Trioxide Aggregate (MTA) has excellent biological property. However, its consistency makes it difficult to be inserted into retrograde cavities. Objective:To evaluate the ability of different methods to fill retrograde cavities with MTA. Material and methods: Root canals of thirty single-rooted resin teeth were prepared and filled. After the cut of 3 mm short of apical third, retrograde cavities with 3 mm deep were prepared using an ultrasound device and retrotips (CVD, São José dos Campos, SP, Brazil). The retrograde preparation was evaluate by using an operative microscope (D.F. Vasconcellos, São Paulo, SP, Brazil). The teeth were randomly divided into three groups (n = 10), according to the method: 1) condenser (Trinity, São Paulo, SP, Brazil); 2) MTA applicator (Angelus, Londrina, Brazil) + condenser; 3) condenser associated with ultrasound (CVD, São José dos Campos, SP, Brazil). After the filling of retrograde cavities with white MTA (Angelus, Londrina, Brazil), teeth were radiographed using a digital system (Kodak RVG 6000, Rochester, NY, USA). The images were analyzed by UTHSCSA Image Tool 3.0 software. The percentage of filling was calculated by the proportion between the total area of retrograde cavity and the filled area. The radiographic density mean of each third of retrograde cavity filled with MTA was measured by using the histogram tool of the software. The results were submitted to ANOVA and Tukey tests, with 5% of significance. Results: There was no difference in percentage of filling among the groups (p > 0.05) (approximately 85%). By comparing the thirds, the condenser and MTA applicator groups showed higher density for apical and middle third than cervical third (p < 0.05). The ultrasound group presented similar density among the thirds. Conclusion: The filling ability was similar for the studied methods. Ultrasound promoted better distribution of MTA in retrograde cavity, but did not increase the density of material.

Estudo da migração de células peritoneais na paracoccidioidomicose experimental murina: avalaição do efeito da estimulaçao de células peritoneais por lectina sobra a evolução da lesão paracoccidióidica

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Paracoccidioides brasiliensis interferes on dendritic cells maturation by inhibiting PGE(2) production

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

Suplementos múltiplos para novilhas de corte a pasto no período seco: características nutricionais

The objective was to evaluate levels of multiple supplements supplied to beef heifers in low supply of pasture on intake and nutrients digestibility, pH and concentration of ruminal ammonia nitrogen, nitrogen balance, synthesis and microbial efficiency. Five Nellore heifers with initial body weight of 344.00 +/- 21.10 kg and average initial age of 22 months were used. The following supplements were evaluated as fed to the animals: control (mineral supplement) and supplementation levels of 2, 4, 6 and 8 kg/animal/day, composing the following percentages of body weight (BW): 0.55, 1.11, 1.66 and 2.22%, respectively. The experimental design was a Latin square with five replications and five supplements. Intakes of total dry matter, crude protein (CP) and total digestible nutrients, and total apparent digestibility of dry matter, crude protein, total carbohydrates and no fiber carbohydrates increased linearly (P <0.0001) and dry matter intake of forage decreased (P <0.0001) with supplementation levels. Supplementation strategies promoted reduction on ruminal pH (P = 0.0343) and increased concentration of ruminal ammonia nitrogen (NH3-N) (P = 0.0004) before the supply of supplements, and after, this NH3-N presented quadratic behavior (P = 0.0023). Nitrogen intake, excretion of urea in urine and nitrogen excretion in feces increased linearly (P <0.0001) according to supplementation levels. The supply of high levels of multiple supplements reduces forage intake and presents itself as an alternative to cattle maintained in a systems with low supply of pasture in the dry season.

Importance of adhesins in virulence of Paracoccidioides spp.

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

Recovery Tests of an Hybrid Current Limiter Composed of a SFCL in Parallel to an Air-Core Power Reactor

The resistive-type superconducting fault current limiters (RSFCL) prototypes using YBCO-coated conductors have shown current limitation for medium voltage class applications for acting time up to 80 ms. By connecting an air-core reactor in parallel with the RSFCL, thus making an hybrid current limiter, one can extend the acting time for up to 1 s. In this work, we report the performance of a hybrid current limiter subjected to an AC peak fault current of 2 kA during 1 s for which within the first 80 ms the SFCL limits the current concurrently with the air-core reactor, and for the remaining 920 ms, only the air-core reactor limits the current. In order to evaluate the actual conditions for subsequent reconnection of RSFCL to the power grid, the hybrid fault current limiter was tested varying the time interval for recovery from 900 ms and 1.2 s, followed again by the concurrent operation of the hybrid limiter during 1 s (SFCL during 80 ms). From this evaluation test, the recovery time can be measured and compared using the voltage peak generated in superconducting module from the first and second fault test. The recovery time was also determined through the pulsed current method (PCM) on short-length sample test. The results showed that the fault current was limited from 1.9 kA down to 514 A after 1 cycle of 60 Hz frequency, with recovery time lower than 1.2 s for two subsequent fault current tests.

Evaluation of the double agar gel immunodiffusion test and of the enzyme-linked immunosorbent assay in the diagnosis and follow-up of patients with chronic pulmonary aspergillosis

Coordenação de Aperfeiçoamento do Pessoal de Nível Superior (CAPES)