Leishmanicidal activity of the venoms of the Scorpions Brotheas amazonicus and Tityus metuendus.

Leishmaniasis is a vector-transmitted zoonosis caused by different species of the genus Leishmania, with a wide clinical spectrum. It is a public health problem aggravated by a series of limitations regarding treatment. In the search for new therapeutic alternatives, scorpion venoms are a source of multifunctional molecules that act against the natural resistance of pathogens. This work evaluated the antileishmanial potential of Brotheas amazonicus and Tityus metuendus venoms against the promastigote forms of Leishmania amazonensis e Leishmania guyanensis. The venoms of B. amazonicus and T. metuendus were evaluated for their constituents using Fourier Transform Infrared (FTIR). Growth inhibition and death of promastigotes were evaluated in the presence of diferente crude venom concentrations (100 µg/mL, 50 µg/mL, 10 µg/mL, 1 µg/mL) after one hour of incubation at 25 °C. The FTIR spectra of both venoms exhibited bands in approximate regions, revealing that both exhibit similar functional groups. Crude venom from both scorpion species showed similar or superior leishmanicidal effects to the standart drug N-methylglucamine antimoniate. At the highest concentration of 100 µg/mL, cultures of L. guyanensis treated with the venom of B. amazonicus showed the highest mortality percentages, above 28%, while T. metuendus venom showed the highest activity against L. amazonensis, with mortality above 7%. This preliminar study demonstrates that B. amazonicus and T. metuendus venoms can be important tools in the search for new drugs Against leishmaniasis. Next step involves evaluating the activity against the amastigote forms and purifying the venom proteins in order to identify the best anti-leishmania candidates.

Expressão gênica em ovócitos suínos de diferentes classificações morfológicas / Gene expression in swine oocytes in different morphological classifications

A produção in vitro de embriões suínos tem alcançado resultados insatisfatórios: ovócitos maturados in vivo produzem uma porcentagem maior de embriões em relação aos maturados in vitro. O sucesso da maturação in vitro está diretamente relacionado com a competência ovocitária. Somente ovócitos competentes são capazes de serem fecundados e terem desenvolvimento embrionário normal. A competência ovocitária pode ser avaliada por vários parâmetros. Recentemente têm sido utilizados como parâmetro os estudos da expressão de genes associados com a competência. O presente trabalho teve por objetivo avaliar diferenças na expressão dos genes BMP15, RYBP, MATER e ZAR1 em ovócitos imaturos de diferentes classes morfológicas, sendo elas: 1, 2, 3 e 4, com a finalidade de proporcionar importantes marcadores moleculares relacionados com a capacidade ovocitária. O RNA total dos ovócitos foi extraído e utilizado como molde para a síntese da primeira fita de cDNA. Os resultados da expressão gênica foram analisados utilizando-se modelo misto, considerando os dados de expressão gênica variável dependente e as classes ovocitárias variáveis independentes. Os genes BMP15, ZAR1 e RYBP apresentaram expressão semelhante nas classes ovocitárias 1, 2 e 3; somente a categoria 4 diferiu na expressão desses genes (P<0,05). O gene MATER foi expresso de forma semelhante em todas as classes ovocitárias estudadas (P>0,05). A técnica de RT-qPCR foi eficiente para detecção desses transcritos em ovócitos de diferentes classes. No entanto, para melhor entendimento do envolvimento desses transcritos na aquisição da competência ovocitária, são necessários mais estudos avaliando ovócitos de diferentes classes morfológicas, em diferentes fases de desenvolvimento, e implicação de outros genes envolvidos com a competência ovocitária.

The in vitro production of pig embryos has achieved unsatisfactory results; in vivo matured oocytes produce a higher percentage of embryos compared to in vitro maturation. The success of in vitro maturation is directly related to oocyte competence. Only competent oocytes are capable of being fertilized and have normal embryonic development. The oocyte competence can be assessed using several parameters. Recently these parameters have been used for gene expression studies associated with competence. This work aimed to evaluate differences in gene expression BMP15, RYBP, MATER, ZAR1 as endogenous control and the constitutive gene GAPDH in immature oocytes of different morphological classes which are: 1, 2, 3 and 4, in order to provide significant molecular markers linked to the ability of development. Oocytes Total RNA was extracted and used as a template for synthesis of the first cDNA strand. The results of gene expression were analyzed using a mixed model, considering the dependent gene expression data and independent ovocitary variable classes. The genes BMP15, RYBP ZAR1 and showed similar ovocitary expression in classes 1, 2 and 3 differ only in category 4 in their expression (P<0.05). The MATER gene was similarly expressed in all ovocitary classes studied (P>0.05). The RTQ-PCR technique was effective for detection of these transcripts in oocytes from different classes. However, for better understanding of the involvement of these transcripts in the acquisition of oocyte competence more studies are needed to evaluate different morphological classes of oocytes at different stages of development and the implication of other genes involved in oocyte competence.

Ultrassonografia testicular em caprinos jovens da raça Alpina / Testicular ultrasonography in young Alpine goats

Objetivou-se com este trabalho observar as mudanças do parênquima testicular de acordo com o fotoperíodo e com a faixa etária dos animais por meio da ecotextura testicular e a viabilidade do uso do ultrassom na avaliação testicular. Foram utilizados 4 animais desde o 6° até o 13° mês de idade. Os exames ecográficos foram realizados a cada 15 dias para avaliar a ecotextura testicular. O parênquima testicular mostrou-se moderadamente ecogênico e homogêneo. Foi observado comportamento semelhante na ecotextura testicular durante o período de avaliação, sendo maior no 8º mês de idade, quando o volume testicular também foi maior; tais mudanças foram relacionadas com o fotoperíodo da região. A avaliação ultrassonográfica dos testículos mostrou-se um método eficiente, capaz de detectar as mudanças no parênquima testicular ao longo do tempo, e como ferramenta alternativa para diferenciação de condições normais e patológicas em caprinos...

The purpose of this study was to observe changes in the testicular parenchyma of male goats. Changes were observed in terms of goat age, evaluating the echotexture and ultrasound viability in the testicles. Four animals were used, ranging from six to 13 months of age. The echographic examinations were performed every 15 days to assess echotexture of the testicle. The testicular parenchyma appeared moderately echogenic and homogeneous. Similar results were observed in testicular echotexture during the evaluation period. The highest results appearing during the eighth month of age, when the testicular volume was also highest; such changes were related to the regional photoperiod. The ultrasound evaluation of the testicles was shown to be an effective method, ultimately capable of detecting changes in testicular parenchyma over time, being an alternative to differentiation normal and pathological conditions in goats...

Efeito da temperatura das amostras de leite na concentração de cálcio solúvel e de beta-caseína: interferência no teste de estabilidade frente ao etanol / Effect of milk sample temperature on the concentration of soluble calcium and beta-casein: interference with the ethanol stability test

O objetivo deste trabalho foi avaliar a variação do perfil proteico e do cálcio solúvel na coagulação do leite pelo etanol nas temperaturas de 4ºC, 10ºC, 15ºC e 20ºC. Amostras de leite de 61 animais foram avaliadas quanto à estabilidade ao etanol nas concentrações de 66 a 92 por cento (v/v) nas temperaturas de 4ºC, 10ºC, 15ºC e 20ºC. Três amostras, após 24 horas de armazenamento a 4ºC, foram ultracentrifugadas em quadruplicata (40.000 x g) a 4ºC e a 20ºC, respectivamente, por 60 minutos. Em seguida, o sobrenadante foi retirado e submetido à análise do cálcio solúvel pela técnica via úmida (digestão nitroperclórica) e leitura em espectrofotômetro de absorção atômica. O perfil proteico foi analisado pela técnica de eletroforese capilar empregando kit específico para determinação proteica. Os resultados mostraram uma correlação positiva entre o aumento da temperatura das amostras e a estabilidade do leite frente às diferentes concentrações de etanol. A porcentagem de cálcio solúvel no sobrenadante após ultracentrifugação foi maior nas amostras tratadas a 4ºC (P<0,05). As amostras ultracentrifugadas na temperatura de 4ºC apresentaram quantidades superiores de β-caseína no sobrenadante em comparação com as amostras tratadas a 20ºC. O abaixamento da temperatura favoreceu a migração da β-caseína e do cálcio coloidal para a fase solúvel do leite, o que possivelmente favoreceu o aumento da instabilidade das amostras no teste do etanol. Os resultados sugerem que a temperatura ideal para a realização de teste de estabilidade do leite frente ao etanol deveria ser de 21ºC...

The aim of this study was to evaluate the variation in protein profile and soluble calcium in milk coagulation by ethanol at 4ºC, 10ºC, 15ºC and 20ºC. Milk samples from 61 dairy cows were evaluated for stability of ethanol concentrations from 66 to 92 percent (v/v) at temperatures of 4°C, 10°C, 15°C and 20°C. Three samples were ultracentrifuged (40,000 x g) after 24 hours of storage at 4°C and 20°C, respectively, for 60 minutes. Their supernatants were removed and subjected to analyses of soluble calcium through nitro-perchloric digestion and atomic absorption spectrophotometry. The protein profiles were determined by capillary electrophoresis using a specific kit for protein determination. The results showed a positive correlation between the increase in temperature of the samples and the stability of milk against various concentrations of ethanol. The percentage of soluble calcium in the supernatant after centrifugation was higher in samples treated at 4°C (P<0.05). The samples ultracentrifuged at 4°C showed higher amounts of β-casein in the supernatant compared with samples stored at 20°C. The lowering of the temperature favored the migration of β-casein and colloidal calcium to the soluble phase of milk, which may also have favored the instability of milk in the ethanol test. According to the results, the milk sample temperature for the ethanol stability test should be 21ºC...

Non-specific effects of the MEK inhibitors PD098,059 and U0126 on glutamate release from hippocampal synaptosomes.

In order to investigate a role for the extracellular-signal-regulated kinases 1 and 2 (ERK1/2) on hippocampal neurotransmitter release, we studied the effect of commonly used MEK (mitogen-activated protein kinase [MAPK]/ERK kinase) inhibitors, PD098,059 and U0126, on depolarization-induced glutamate release. PD098,059 inhibited glutamate release from hippocampal synaptosomes stimulated with 15 mM KCl in a concentration-dependent manner. At the same range of concentrations, PD098,059 inhibited basal and KCl-stimulated ERK1/2 phosphorylation. U0126, however, did not significantly affect KCl-evoked glutamate release at concentrations shown to inhibit ERK activity. Nonetheless, U0126 unspecifically potentiated depolarization-induced Ca2+-independent glutamate release, which masked a small dose-dependent inhibitory effect on the Ca2+-dependent release. PD098,059 reduced the [Ca2+]i response to KCl by partially inhibiting Ca2+ entry through N- and P-/Q-type voltage-gated Ca2+ channels, whereas U0126 did not affect depolarization-induced Ca2+ influx. To overcome the unspecific effect of PD098,059 on Ca2+ entry, we studied the effect of both MEK inhibitors on glutamate release stimulated by a Ca2+ ionophore. PD098,029 and U0126 showed a small dose-dependent inhibitory effect on ionomycin-induced glutamate release, at concentrations shown to inhibit ionomycin-stimulated ERK phosphorylation. These findings uncover new unspecific actions for both MEK inhibitors and suggest a minor role for ERK in modulating glutamate release in the hippocampus.

Role of Pasteurella granulomatis and Dermatobia hominis in the etiology of lechiguana in cattle.

Attempts were made to reproduce bovine lechiguana, a disease associated with Dermatobia hominis and Pasteurella granulomatis infections. Suspensions of Pasteurella granulomatis were mixed with each of the following: saponin, oil adjuvant, ground Dermatobia hominis, or 5% mucin. Each preparation was inoculated into 6 cattle. Twelve more cattle, 6 of which received dexamethasone, were inoculated with bacterial suspension alone. Abscesses but no lechiguana was produced in all 36 cattle. After abscess regression, 12 cattle were reinoculated with a suspension of mouse-passed P. granulomatis. Only abscesses were produced. The intralymphatic inoculation of P. granulomatis in 6 cattle did not produce the disease. Eleven cattle infected naturally with D. hominis had lesions containing dead larvae. These lesions were inoculated with P. granulomatis. Nine cattle were experimentally infected with larvae of D. hominis that had been contaminated with the bacteria. No lechiguana lesions were produced in these 20 cattle. Six cattle with severe natural D. hominis infection were inoculated in the larval lesions with P. granulomatis. One developed lesions indistinguishable from those of natural lechiguana. The lesions regressed after treatment with chloramphenicol. D. hominis larvae and exudate from lesions caused by the fly were collected from 7 cattle on 3 farms and examined bacteriologically. P. granulomatis was isolated from the larvae and the exudate of a healthy calf from a farm where lechiguana had never been observed. These results suggest that P. granulomatis has a causal role in lechiguana, and that D. hominis may be a carrier of the bacterium. These observations suggest that lechiguana occurs when severe D. hominis lesions are infected with P. granulomatis. The apparent long incubation period, the negative results obtained in the other experiments, and also the infrequent occurrence of the natural disease suggest that lechiguana is a disease for which Koch's postulates are not easily fulfilled.

Efeitos da radiaçäo soft-laser (diodo) sobre o processo de cicatrizaçäo cutânea em felinos / The effect of soft-laser (diodes) radiation on wound healing in cats

Verificou-se o valor do emprego do laser diodo Ga-As-SHLD para auxiliar o processo de cicatrizaçäo cutânea em felinos. Utilizaram-se 21 animais que foram submetidos à ovariohisterectomia, 9 pertencendo ao grupo controle e 12 submetidos à laserterapia. Os exames anátomopatológicos foram realizados nos dias 2, 4 e 8 de pós-operatório, juntamente com avaliaçäo macroscópica. Concluiu-se que a aplicaçäo do laser diodo Ga-As-SHLD auxiliou o processo cicatricial da ferida cirúrgica por aumento de resistência à tensäo

[Utilization of the Kato-Katz-method (thick-smear technique) the diagnosis of Isospora (author's transl)]. / Utilização do método de Kato-Katz para o diagnóstico de Isospora; resultados preliminares.

Performing 10475 stool examination using the KATO-KATZ, HOFFMAN-PONS-JANER and direct examination (after agitation) methods, the author showed the best results with KATO-KATZ method for the diagnosis of Isospora hominis and Isospora belli. Among the 10475 stools examined 392 (3.74%) were positive for I. hominis and 55 (0.52%) for I. belli with the KATO-KATZ method; with the use of the others methods only 28 (0.26%) were positive for I. hominis and 6 (0.06% for I. belli.