The ""single-section"" Golgi method adapted for formalin-fixed human brain and light microscopy

The Golgi method has been used for over a century to describe the general morphology of neurons in the nervous system of different species. The ""single-section"" Golgi method of Gabbott and Somogyi (1984) and the modifications made by Izzo et al. (1987) are able to produce consistent results. Here, we describe procedures to show cortical and subcortical neurons of human brains immersed in formalin for months or even years. The tissue was sliced with a vibratome, post-fixed in a combination of paraformaldehyde and picric acid in phosphate buffer, followed by osmium tetroxide and potassium dicromate, ""sandwiched"" between cover slips, and immersed in silver nitrate. The whole procedure takes between 5 and 11 days to achieve good results. The Golgi method has its characteristic pitfalls but, with this procedure, neurons and glia appear well-impregnated, allowing qualitative and quantitative studies under light microscopy. This contribution adds to the basic techniques for the study of human nervous tissue with the same advantages described for the ""single-section"" Golgi method in other species; it is easy and fast, requires minimal equipment, and provides consistent results. (C) 2010 Elsevier B.V. All rights reserved.

A technique for preserving pigmentation in some capsalid monogeneans for taxonomic purposes

A technique is described to preserve the pigment found in the bodies and the intestine of some brightly coloured and darkly pigmented benedeniine capsalid monogeneans. Previous studies of these pigmented capsalids have proven difficult because the pigmentation usually disappears when the worms are fixed using preservatives containing concentrations of formalin over 5% and/or ethanol, acetic acid, chromic acid, picric acid and mercuric chloride. The technique developed here uses a fixative comprising glycerol, acetone and formalin (GAF). After fixation under light coverslip compression for three minutes, specimens are transferred to absolute acetone for three minutes and cleared in a mixture of nine parts cedar wood oil and one part absolute acetone before mounting in Canada balsam. Processing must be carried out quickly, as these chemicals will cause the pigments to fade if the specimens are exposed to them for too long. Pigmented benedeniines processed using this technique retain the distribution, intensity and colour observed in live worms. The colour and distribution of pigmentation in monogeneans may be of taxonomic importance and this technique aids preparation of whole-mounts suitable for registration as type-material.

Síntese, aplicação e avaliação da toxicidade aguda de complexo orgânico à base de európio proposto como marcador fotoluminescente para a Identificação de resíduos de tiro

Um complexo de alta fotoluminescência é proposto como marcador óptico para a identificação de resíduos de tiro (GSR). O marcador é o complexo [Eu(PIC)3(NMK)3], de fórmula molecular Eu(C6H2N3O7)3.(C7H13NO)3, que apresenta o íon Eu3+ e os ligantes ácido pícrico (PIC) e n-metil-Ɛ-caprolactama (NMK). Foi realizada a caracterização quimicamente através de espectroscopia de emissão, espectroscopia de infravermelho com transformada de Fourier (FTIR), termogravimetria e análise térmica diferencial (TG/DTA), e espectrometria de massas com ionização por eletrospray e ressonância ciclotrônica de íons por transformada de Fourier (ESI-FT-ICR MS), e, em seguida, foram adicionadas diferentes massas do complexo a munições convencionais (de 2 a 50 mg por cartucho). Após os tiros, o GSR marcado foi visualmente e quimicamente detectado por irradiação UV (ʎ = 395 nm) e ESI-FT-ICR MS, respectivamente. Os resultados mostraram uma fotoluminescência eficiente e duradoura, sendo facilmente visível sobre a superfície do alvo, no ambiente, no cartucho deflagrado, na arma de fogo, e sobre as mãos e braços do atirador quando utilizada massa a partir de 25 mg do marcador em cartuchos .38 e 50 mg em cartuchos .40. Sua toxicidade aguda também foi avaliada empregando-se o Protocolo 423 da Organização para a Cooperação e Desenvolvimento Econômico (OECD) e apresentou DL50 de 1000 mg.kg-1, sendo classificado como de categoria 4 na escala do Sistema Globalmente Harmonizado de Classificação e Rotulagem de Produtos Químicos (GHS), considerado, portanto, de média toxicidade. O composto mostrou ser menos tóxico do que os componentes inorgânicos de munições convencionais (em especial o Pb), justificando o seu emprego como marcador de GSR.

Substituted P-Phenylene Ethynylene trimers as fluorescent sensors for nitroaromatic explosives

New sensory materials based on p-phenylene ethynylene trimers integrating calix[4]arene receptors (CALIX-PET) and tert-butylphenol (TBP-PET) moieties have been synthesized and their sensitivity and selectivity for the detection of nitroaromatic compounds (NACs) such as nitrobenzene (NB), 2,4-dinitrotoluene (2,4-DNT), 2,4,6-trinitrotoluene (TNT) and picric acid (PA) investigated in fluid phase and solid-state. It was found that both fluorophores displayed high sensitivities toward NACs detection in solution as evaluated by the Stern-Volmer formalism. For all the tested explosives, the ratio of fluorescence intensities (F-0/F) is a linear function of the quencher concentration only after appropriate correction of fluorescence quenching data for inner-filter effects. The quenching efficiencies for CALIX-PET and TBP-PET follow the order PA >> TNT > DNT > NB, which correlate well with the quenchers electron affinities as evaluated from their LUMOs energies thereby suggesting a photoinduced electron transfer as the dominant mechanism of fluorescence quenching. The selectivity of these sensors was checked against exemplar interferents possessing differentiated electronic properties (benzoic acid, 2,4-dichlorophenol and benzoquinone) and reduced quenching activity was detected. The quenching efficiencies and response times of the two fluorophores in the solid-state toward NB, 2,4-DNT and TNT vapors were evaluated through steady-state fluorescence quenching experiments with the materials dispersed in polymeric matrices or as neat films. The most significant fluorescence quenching responses were achieved for drop-casted films of TBP-PET upon exposure to nitroaromatics.

Novos polímeros conjugados baseados em calixarenos contendo fontes quirais de origem natural: síntese, propriedades e aplicações sensoriais

Trabalho Final de Mestrado para obtenção do grau de Mestre em Engenharia Química e Biológica

Caracterização mecânica e microestrutural de um aço baixo carbono microligado com estrutura multifásica

Neste trabalho foi realizada a caracterização mecânica e microestrutural de um aço microligado com estrutura multifásica. Foi aplicado tratamento térmico pré-determinado, objetivando a formação de uma microestrutura multifásica no material. Na caracterização microestrutural foram utilizados ataques químicos à base de metabissulfito de sódio e ácido pícrico, enquanto a caracterização mecânica foi realizada através de ensaios de tração. Os resultados demonstram o elevado potencial dos aços multifásicos em aplicações que necessitem de valores superiores de resistência e ductilidade, pois tanto para temperatura isotérmica de 400ºC quanto para 350ºC houve um ganho no limite de resistência à tração ficando em torno de 786MPa e 773MPa respectivamente, representando um aumento de 15,5% e 13,6% com relação ao material fornecido.

Observações clínicas, anatomopatológicas e tratamento de seis casos de dermovilite exsudativa vegetante crônica (cancro da ranilha) de cqüinos

Chronic hypertrophic pododermatitis cases in six horses of different breeds, aging 14 months to 19 years are described. The lesion begun with a infiltrative tissue in the frog and sole regions of the hoof, characterized by a fast and disorganized growth, with a papillary aspect, white colored in the roots and dark on the extremity, with a necrotic secretion and an extremely fetid odor. Microscopically, an exuberant epidermic proliferative tissue was observed, intermingled with little connective tissue. The horses were divided into two treatment groups. In the first group, including three young mares and a foal, showing lesions in only one limb, a surgical resection of the invasive mass was performed, followed by cauterization of the remaining edges and subsequent daily local application of antiseptic substances. In three of these horses, recurrence of the initial lesion occurred, with fast growth of hyperplasic tissue, affecting almost all the frog and half of the sole. Two horses developed contraction deformities of the hoof. In the second group, one male and one female, each with lesions in two limbs, after surgical debridement of the tissue, the animals received daily applications of picric acid 5%, associated to local use of oxitetracyclin. Although one of these cases required a second surgical intervention for removal of the mass, the horses showed after a period of two to three months total absence of the infiltrative tissue. The use of local picric acid 5% and oxitetracyclin associated to previous surgical debridement showed to be more efficient than the use of antiseptic substances in the treatment of chronic hypertrophic pododermatitis.

Free radical scavenging activity of Kielmeyera variabilis (Clusiaceae)

As part of our ongoing research on antioxidant agents from Brazilian flora, we screened the free radical scavenging activity of two extracts and eight fractions of Kielmeyera variabilis (Clusiaceae) using DPPH (2,2-diphenyl-1- picrylhydrazyl-hydrate) and ABTS+ [2,2'-azinobis(3- ethylenebenzothiazoline-6-sulfonic acid)] colorimetric assays. The ethyl acetate and n-butanol fractions of the leaves of K. variabilis displayed the strongest activity (IC50 of 3.5 ± 0.3 and 4.4 ± 0.2 μg mL -1 for DPPH and 6.6 ± 0.4 and 3.1 ± 0.1 μg mL -1 for ABTS+, respectively). Chromatographic fractionation of the most potent fractions led to identification of three flavonols with previously described antioxidant activity, quercitrin (1), quercetin-3-O- glucoside (3), and quercetin-3-O-galactoside (4), and of one biflavone, podocarpusflavone A (2). This is the first time that the presence of these flavonoids in Kielmeyera variabilis has been reported. © 2013 by the authors.

Transporte placentário via cavéola na placenta de bovinos clonados e transgênicos

Pós-graduação em Ciência e Tecnologia Animal - FEIS

Caracterização das proteínas caveolinas -1 e -2 na placenta de conceptos bovinos clonados transgênicos

The transgenic application of green fluorescent protein (GFP) as fetal cell marker on cattle cloned placenta could provide an exclusive model for studying the morphologic and immunologic maternal-fetal interactions, providing information about its mapping, distinguishing the fetal from maternal cells. This model will have direct application, mainly because these animals present problems during its development. With this model's support, we intend to verify the substances transport between mother and fetus during endocytosis, through the immunolocalization of protein named caveolae. For these, we used 06 cloned bovine and 30 cattle samples of artificial insemination (AI) with 90 days of pregnancy, which had been their development interrupted by humanitarian slaughter of the recipient and recovery of the pregnant uterus. We collected the placentome and the chorion. A part of the samples was cut and fixed, by immersion, on a solution containing 4% of parafomaldehyde or 10% of formaldehyde on a sodium phosphate buffer (PBS), at 0,1 M pH 7.4, Zamboni solution (4% of paraformaldehyde, 15% of picric acid, on sodium phosphate buffer 0,1 M pH 7.4), metacarn (60% of metanol, 30% of chloroform, and 10% glacial acetic acid), for morphologic and immunohistochemistry verification for caveolinas proteins -1 and -2 (CAV -1 and CAV-2). The caveolins -1 were found in fetal and maternal villi, but its strongest staining was observed in the endometrial stroma. The caveolins -2 had positive staining in trophoblast and chorioallantoic membrane, and specifically in giant trophoblastic binucleated cell. Therefore the results were compared between cloned cattle and from AI or natural mating, for assisting on detection of the reason of many placental alterations, embryonic losses, spontaneous abortion, post-natal mortality and large offspring syndrome on laboratory-manipulated animals. The result suggests that the proteins caveolins -1 and -2 (CAV-1 and CAV-2) are part of the caveolae composition and important structures related to the molecule transfer to the fetus, nourish it through endocytosis and pinocytosis.

Determinação simultânea de biomarcadores de função renal em urina utilizando dispositivo analítico microfluídico em papel

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Avaliação farmacognóstica qualitativa das folhas do vegetal Bambusa textilis

Introduction: Bamboos belong to the family Graminae, Bambusoidae subfamily, represented by about 1,250 species worldwide. Originally employed in construction and power, are currently the subject of investigations related to its therapeutic properties in neoplasms. The main species used for therapeutic purposes based on popular knowledge are: Phyllostachys nigra; Bambusa breiflora; tuldoides Bambusa textilis and Bambusa. The literature on the therapeutic action of bamboo species is scarce, but recent studies report a promising effect in the treatment of cancer and other chronic diseases. Objective: This study aimed to evaluate qualitatively the phytochemical composition of plant extract obtained from the leaves of the bamboo species Bambusa textilis, comparing this composition from vegetable leaves with 18 and 24 months of age. Methodology: After collecting plant leaves with 18 or 24 months, they were identified and submitted to drying and milling. For qualitative analysis of its components were employed methods of macroscopic evaluation (mucilage), method of benzoin (resin), reaction Shinoda (flavonoids), reaction with gelatin solution (tannins), boiling and foaming (saponins) and jobs of reactive Wagner, Bertrand, Dragendorff, Mayer, picric acid and tannic acid (alkaloids) (Biavatti; MILK, 2007). Results: in the youngest leaves were found positive for alkaloids, flavonoids, resins and saponins. In the leaves of the plant with 24 months were found only alkaloids flavonoids and resins. Conclusion: the composition of vegetable substances were found associated with a significant therapeutic potential difference and the phytochemical composition in comparison with plant leaves 18 or 24 months. Additional studies are needed to quantify these components, as well as the clarification of its action in the fight against cancer and chronic diseases.

Development of an enzymeless electroanalytical method for the indirect detection of creatinine in urine samples

The aim of this study is to develop a new enzymeless electroanalytical method for the indirect quantification of creatinine from urine sample. This method is based on the electrochemical monitoring of picrate anion reduction at a glassy carbon electrode in an alkaline medium before and after it has reacted with creatinine (Jaffe's reaction). By using the differential pulse voltammetry technique under the optimum experimental conditions (step potential, amplitude potential, reaction time, and temperature), a linear analytical curve was obtained for concentrations of creatinine ranging from 1 to 80 mu mol L-1, with a detection limit of 380 nmol L-1. This proposed method was used to measure creatinine in human urine without the interference of most common organic species normally present in biological fluids (e.g., uric acid, ascorbic acid, glucose, and phosphocreatinine). The results obtained using urine samples were highly similar to the results obtained using the reference spectrophotometric method (at a 95% confidence level). (C) 2012 Elsevier B.V. All rights reserved.

The Crosslinking Degree Controls The Mechanical, Rheological, And Swelling Properties Of Hyaluronic Acid Microparticles.

Viscosupplements, used for treating joint and cartilage diseases, restore the rheological properties of synovial fluid, regulate joint homeostasis and act as scaffolds for cell growth and tissue regeneration. Most viscosupplements are hydrogels composed of hyaluronic acid (HA) microparticles suspended in fluid HA. These microparticles are crosslinked with chemicals to assure their stability against enzyme degradation and to prolong the action of the viscosupplement. However, the crosslinking also modifies the mechanical, swelling and rheological properties of the HA microparticle hydrogels, with consequences on the effectiveness of the application. The aim of this study is to correlate the crosslinking degree (CD) with these properties to achieve modulation of HA/DVS microparticles through CD control. Because divinyl sulfone (DVS) is the usual crosslinker of HA in viscosupplements, we examined the effects of CD by preparing HA microparticles at 1:1, 2:1, 3:1, and 5:1 HA/DVS mass ratios. The CD was calculated from inductively coupled plasma spectrometry data. HA microparticles were previously sized to a mean diameter of 87.5 µm. Higher CD increased the viscoelasticity and the extrusion force and reduced the swelling of the HA microparticle hydrogels, which also showed Newtonian pseudoplastic behavior and were classified as covalent weak. The hydrogels were not cytotoxic to fibroblasts according to an MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay. © 2014 Wiley Periodicals, Inc. J Biomed Mater Res Part A, 2014.

Geochemistry Of Acid Mine Drainage From A Coal Mining Area And Processes Controlling Metal Attenuation In Stream Waters, Southern Brazil.

Acid drainage influence on the water and sediment quality was investigated in a coal mining area (southern Brazil). Mine drainage showed pH between 3.2 and 4.6 and elevated concentrations of sulfate, As and metals, of which, Fe, Mn and Zn exceeded the limits for the emission of effluents stated in the Brazilian legislation. Arsenic also exceeded the limit, but only slightly. Groundwater monitoring wells from active mines and tailings piles showed pH interval and chemical concentrations similar to those of mine drainage. However, the river and ground water samples of municipal public water supplies revealed a pH range from 7.2 to 7.5 and low chemical concentrations, although Cd concentration slightly exceeded the limit adopted by Brazilian legislation for groundwater. In general, surface waters showed large pH range (6 to 10.8), and changes caused by acid drainage in the chemical composition of these waters were not very significant. Locally, acid drainage seemed to have dissolved carbonate rocks present in the local stratigraphic sequence, attenuating the dispersion of metals and As. Stream sediments presented anomalies of these elements, which were strongly dependent on the proximity of tailings piles and abandoned mines. We found that precipitation processes in sediments and the dilution of dissolved phases were responsible for the attenuation of the concentrations of the metals and As in the acid drainage and river water mixing zone. In general, a larger influence of mining activities on the chemical composition of the surface waters and sediments was observed when enrichment factors in relation to regional background levels were used.