Bioresponsive nanostructured systems for sustained naltrexone release and treatment of alcohol use disorder: Development and biological evaluation.

In this study, microemulsions capable of transforming into nanostructured hexagonal phase gels in vivo upon uptake of biological fluids for naltrexone prolonged release were investigated as a strategy for management of alcohol use disorder (AUD). Microemulsions were prepared using monoolein, tricaprylin, water and propylene glycol; after preliminary characterization, one formulation was selected, which contained 55% of monoolein-tricaprylin (M-55). This microemulsion displayed size below 200 nm and Newtonian rheological behavior. Liquid crystalline gels formed in vitro upon 8 h of contact with water following a second order kinetics. After 120 h, <50% of naltrexone was released in vitro independently on drug loading (5 or 10%). In vivo, gels formed within 24 h of M-55 subcutaneous administration, and persisted locally for over 30 days providing slow release of the fluorescent marker Alexa fluor compared to a solution. Using the conditioned place preference paradigm, a test used to measure drug's rewarding effects, a single dose of M-55 containing 5% naltrexone reduced the time spent in the ethanol-paired compartment by 1.8-fold compared to saline; this effect was similar to that obtained with daily naltrexone injections, demonstrating the formulation efficacy and its ability to reduce dosing frequency. A more robust effect was observed following administration of M-55 containing 10% of naltrexone, which was compatible with aversion. These results support M-55 as a platform for sustained release of drugs that can be further explored for management of AUD to reduce dosing frequency and aid treatment adherence.

Perfil eletroforético do colostro de ovelhas suplementadas com propileno glicol e cobalto associado à vitamina B12 no final da gestação / Electrophoretic profile of the colostrum of ewes supplemented with propylene glycol and cobalt associated with vitamin B12 in late pregnancy

Este trabalho teve por objetivo avaliar o proteinograma do colostro de ovelhas submetidas a administração de propileno glicol e de cobalto associado à vitamina B12 no final da gestação. Dezoito ovelhas da raça Santa Inês, prenhas e com idade variando entre 18 meses a cinco anos foram distribuídas, por amostragem probabilística em três grupos experimentais, aproximadamente 30 dias antes da data prevista para o parto. No Grupo 1 (G1/n=6) foram administrados 30mL de propileno glicol P.A. via oral diariamente; no Grupo 2 (G2/n=6) foi administrado 1mg de cloreto de cobalto em solução a 1% via oral diariamente e 2mg de vitamina B12, via intramuscular semanalmente e no Grupo 3 (G3/n=6): grupo controle. Logo após o parto procedeu-se a colheita de 30mL de colostro, que foram acondicionados em recipientes apropriados e encaminhados ao laboratório. Após homogeneização, adicionou-se a cada 1.000µL de colostro, 75µL de solução de renina, que foi mantido em banho-maria a 37ºC por aproximadamente 20 minutos e centrifugado a 21.000G durante dez minutos em centrífuga refrigerada. Posteriormente, a fração intermediária, correspondente ao soro do colostro, foi aliquotada e mantida em ultrafreezer a -80oC para posterior determinação das proteínas. A determinação da proteína total do soro colostral foi realizada empregando-se reagente comercial. A separação das proteínas foi realizada utilizando-se a técnica de eletroforese em gel de poliacrilamida contendo dodecil sulfato de sódio (SDS-PAGE). Foram identificadas as proteínas IgA, lactoferrina, albumina, IgG de cadeia pesada (IgGCP), ß-caseína, IgG de cadeia leve (IgGCL), ß-lactoglobulina and α-lactoalbumina, não havendo influência da administração dos suplementos na fase final da gestação sobre as concentrações protéicas do colostro.(AU)

This study aimed to evaluate the proteinogram of the colostrum of ewes submitted to administration of propylene glycol and cobalt associated with vitamin B12 in late pregnancy. Eighteen pregnant Santa Inês ewes 18 months to 5 years old were distributed by probabilistic sampling into three experimental groups, about 30 days before the expected delivery date. In group 1 (G1/n=6), daily oral doses of 30ml propylene glycol PA were administered; Group 2 (G2/n=2) received a daily oral dosage of 1mg cobalt chloride in 1% solution and 2mg of vitamin B12 intramuscularly once a week, and Group 3 (G3/n=6) was the control group. Soon after delivery 30mL of colostrum was harvested from each ewe, which were stored in appropriate containers and sent to the laboratory. After homogenization, we added to each 1000µL of colostrum 75µL solution of rennin, which was kept in a water bath at 37°C for about 20 minutes and centrifuged at 21.000G for 10 minutes in a refrigerated centrifuge. Later, the intermediate fraction, corresponding to colostrum whey, was aliquoted and kept in a -80oC ultrafreezer for subsequent determination of proteins. The determination of the total colostral protein whey was performed using a commercial reagent, observing the linearity test for colostrum. The separation of proteins was performed using the technique of electrophoresis on polyacrylamide gel containing sodium dodecyl sulfate (SDS-PAGE). Lactoferrin, IgA, albumin, IgG heavy chain (IgGCP), ß-casein, IgG light chain (IgGCL), ß-lactoglobulin and α-lactalbumin proteins were identified. There was no influence of the administration of supplements during late pregnancy on the concentration of proteins identified in the colostrum of the ewes.(AU)

Avaliação in vitro da atividade fotoprotetora de extratos vegetais do cerrado de Mato Grosso / In vitro sunscreen activity evaluation of plants Iextracts from Mato Grosso cerrado

O cerrado brasileiro é um bioma detentor de grande diversidade biológica. No entanto, são escassas as pesquisas de espécies vegetais, especialmente do cerrado mato-grossense, com potencial para serem utilizadas como filtros solares naturais. O objetivo desta pesquisa foi estudar o potencial fotoprotetor de espécies de diferentes famílias (Apocynaceae, Lythraceae, Oxalidaceae) do cerrado da região do Rio Manso, Chapada dos Guimarães - MT. A absorbância dos extratos etanólicos secos foram medidas em diferentes concentrações entre os comprimentos de onda de 260 a 400nm para verificar a absorção nas regiões ultravioleta A e B (UVA e UVB). As plantas que apresentaram absorbância na região estudada foram submetidas a uma análise fitoquímica qualitativa preliminar para determinar a presença de polifenóis e alcalóides, constituintes característicos de plantas que absorvem a radiação UV. Os extratos etanólicos secos que apresentaram absorção em UVB foram submetidos ao teste de determinação in vitro do Fator de Proteção Solar (FPS) desenvolvido por Mansur. M. Velame apresentou absorção na região UVB com absorbância máxima em 318nm, enquanto que a L. pacari e O. hirsutissima apresentaram absorbância na região UVA. Na concentração utilizada e padronizada, nenhuma das espécies apresentou FPS >2, sendo assim não podem ser consideradas plantas com potencial fotoprotetor.

The Brazilian savanna is a holding biome of large biological diversity. However, the researches of plants species are scarce, especially at the Mato Grosso's savanna; which have potential to be used as natural sunscreen. The objective of this research was to study the photoprotector potential of several species (Apocynaceae, Lythraceae, Oxalidaceae) from the savanna's region at the Manso River, Chapada dos Guimarães - MT. The absorbance of dry ethanolic extracts were measured in different concentrations, between waves from 260nm until 400 nm in length. Just to check the absorption in the A and B ultraviolet regions (UVA and UVB). The plants that presented absorbance by the studied area were submitted to a preliminary qualitative phytochemical analysis to determine if there are polyphenols and alkaloids inside, because they are typical constituents of plants that absorber the UV radiation. The dry ethanolic extracts, that presented absorption in UVB, were submitted to a in vitro Sun Protection Factor (SPF) determination test, developed by Mansur. M.velame presented absorption in the UVB region with maximal absorbance at 318 nm, while L. pacari and O. hirsutissima presented absorbance in the UVA region. At the used and standardized concentration, no species presented SPF > 2, so they cannot be considered plants with photoprotector potential.

Aspectos clínicos e características do leite em ovelhas com mastite induzida experimentalmente com Staphylococcus aureus / Clinical aspects and characteristics of the milk in sheep with mastitis experimentally induced with Staphylococcus aureus

Este trabalho teve por objetivo estudar os aspectos clínicos e as características físico-químicas do leite em ovelhas com mastite induzida experimentalmente com Staphylococcus aureus. Foram utilizados dez animais da raça Santa Inês, com peso médio de 30 kg, fêmeas, primíparas recém-paridas, mantidos em apriscos e clinicamente sadios. Após se estabelecer os padrões de normalidade para as variáveis estudadas, os animais foram inoculados experimentalmente numa mama com uma cepa de S. aureus, empregando-se o inóculo de 1,0x10(4)ufc/ml, enquanto a outra serviu como controle. As observações clínicas e laboratoriais foram realizadas nos intervalos de 12, 24, 36, 48, 60, 72, 84, 96, 108, 120, 132, 168, 180, 288 e 336 horas após a inoculação do agente etiológico (PI). Todos os animais apresentaram manifestações clínicas sistêmicas e nas glândulas inoculadas, observadas com mais intensidade a partir de 24 horas após a inoculação. Alterações significativas (P<0,05) na produção e nos componentes físico-químicos do leite em relação aos seus controles ocorreram, reduzindo o volume e gordura. O pH, o teor de cloretos e a CCS atingiram índices muito elevados (P<0,05), acompanhados da reação na prova do CMT. O exame bacteriológico revelou a presença do S. aureus durante a fase de infecção. Após a instituição do tratamento nas ovelhas às 36 h PI, um animal morreu 48h PI; nas demais ocorreu a recuperação clínica, no entanto não houve o restabelecimento fisiológico das mamas inoculadas, que perderam a sua funcionalidade.

The objective was to study the clinical aspects and the physical-chemical characteristics of the milk in sheep with mastitis experimentally induced with Staphylococcus aureus. For such, were used 10 Santa Inês primiparity ewes, weighing 30 kg, clinically healthy and housed in a stall. After establishing the normality patterns for the studied variables, the animals were inoculated into one teat of the udder with an inoculum of 1.0x10(4)ufc/ml of S. aureus, while the other gland served as control. The clinical observations were accomplished in intervals of 12, 24, 36, 48, 60, 72, 84, 96, 108, 120, 132, 168, 180, 288 and 336 hours after the inoculation of the agent (PI). All the animals presented clinical systemic manifestations, and in the inoculated glands with more intensity from 24 hours on after the inoculation. There were significant alterations (P<0.05) in the production and physical-chemical composition of the milk in relation to the controls, with reduction of volume and fat. The pH, the chlorates and CCS reached very high indexes, accompanied by the CMT test reaction. The bacteriological exam revealed the presence of S. aureus during the phase of infection. After the treatment of the sheep 36 hours PI, one animal came to death 48h PI, and the others showed clinical recovery; however there was no physiologic re-establishment of the inoculated udders which lost their function.