Formation of Lipofuscin-Like Autofluorescent Granules in the Retinal Pigment Epithelium Requires Lysosome Dysfunction

PURPOSE: We aim to characterize the pathways required for autofluorescent granule (AFG) formation by RPE cells using cultured monolayers. METHODS: We fed RPE monolayers in culture with a single pulse of photoreceptor outer segments (POS). After 24 hours the cells started accumulating AFGs that were comparable to lipofuscin in vivo. Using this model, we used a variety of light and electron microscopical techniques, flow cytometry and Western blot to analyze the formation of AFGs. We also generated a mutant RPE line lacking cathepsin D by gene editing. RESULTS: AFGs seem to derive from incompletely digested POS-containing phagosomes and after 3 days are surrounded by a single membrane positive for lysosome markers. We show by various methods that lysosome-phagosome fusion is required for AFG formation, and that impairment of lysosomal pH or catalytic activity, particularly cathepsin D activity, enhances AF accumulation. CONCLUSIONS: We conclude that lysosomal dysfunction results in incomplete POS degradation and enhanced AFG accumulation

In Vitro and In Vivo Investigation of the Efficacy of Arylimidamide DB1831 and Its Mesylated Salt Form - DB1965 - against Trypanosoma cruzi Infection

Chagas disease is caused by infection with the intracellular protozoan parasite Trypanosoma cruzi. At present, nifurtimox and benznidazole, both compounds developed empirically over four decades ago, represent the chemotherapeutic arsenal for treating this highly neglected disease. However, both drugs present variable efficacy depending on the geographical area and the occurrence of natural resistance, and are poorly effective against the later chronic stage. As a part of a search for new therapeutic opportunities to treat chagasic patients, pre-clinical studies were performed to characterize the activity of a novel arylimidamide (AIA - DB1831 (hydrochloride salt) and DB1965 (mesylate salt)) against T.cruzi. These AIAs displayed a high trypanocidal effect in vitro against both relevant forms in mammalian hosts, exhibiting a high selectivity index and a very high efficacy (IC50 value/48 h of 5–40 nM) against intracellular parasites. DB1965 shows high activity in vivo in acute experimental models (mouse) of T.cruzi, showing a similar effect to benznidazole (Bz) when compared under a scheme of 10 daily consecutive doses with 12.5 mg/kg. Although no parasitological cure was observed after treating with 20 daily consecutive doses, a combined dosage of DB1965 (5 mg/kg) with Bz (50 mg/kg) resulted in parasitaemia clearance and 100% animal survival. In summary, our present data confirmed that aryimidamides represent promising new chemical entities against T.cruzi in therapeutic schemes using the AIA alone or in combination with other drugs, like benznidazole

Conduta nutricional de bailarinos adolescentes de ambos os sexos

A balanced diet is a major factor for dancers because of their contribution to improving the body composition and physical fitness of this audience. The objective of the research was to evaluate the nutritional behavior of macro- and micronutrients in the diet of classic adolescent dancers. Nine male dancers of 16.4 ± 1.5 years old and female 14.5 ± 0.9 years old were evaluated. The fat percentage showed values of 8.8 ± 1.3% and 8.7 ± 2.2% for boys and girls in this order. The food consumption of the dancers was recorded in the semi-quantitative food frequency questionnaire. Boys rarely consumed milks and derivatives 28.8%, vegetables and fruits 70%, beverages 35.7% and products diet or light 100% where p˂0.05. Girls consumed snacks and canned once a week 66.7% and rarely drinks 84.6% and diet light products 92.9% where p˂0.05. Macronutrient intake was determined through a three-day dietary recall. The percentage of carbohydrate in the diet of the dancers was 57.8 ± 16.1%, of the dancers 59.7 ± 5.5%. For the protein the value for the dancers was 18 ± 6.2% and 17 ± 2.9 for the dancers. Considering lipids, the ingested numbers were 19.6 ± 7.6% and 21.7 ± 4.7% in this order for boys and girls where, p≥0.05. In conclusion, the dancers of the present study showed a feeding inadequacy, not consistent with the daily nutrient requirements for their dance modality.Uma alimentação balanceada constitui um fator primordial para os bailarinos, devido a sua contribuição para melhorar a composição corporal e aptidão física deste público. O objetivo da pesquisa foi avaliar a conduta nutricional de macro- e micronutrientes na dieta de bailarinos clássicos adolescentes. Foram avaliados nove bailarinos, do sexo masculino com 16,4 ± 1,5 anos e feminino 14,5 ± 0,9 anos. O percentual de gordura mostrou valores de 8,8 ± 1,3% e 8,7 ± 2,2% para os meninos e meninas nesta ordem. O consumo alimentar dos bailarinos, foi registrado no questionário semi-quantitativo de frequência alimentar. Os meninos raramente consumiam leites e derivados 28,8%, hortaliças e frutas 70%, bebidas 35,7% e produtos diet ou light 100% onde p˂0,05. As meninas consumiam petiscos e enlatados 1 vez por semana 66,7% e raramente bebidas 84,6% e produtos diet light 92,9% onde p˂0,05. A ingesta de macronutrientes foi determinada por intermédio de um recordatório alimentar de três dias. O percentual de carboidrato ingerido na dieta dos bailarinos foi de 57,8 ± 16,1% e das bailarinas de 59,7 ± 5,5%. Para as proteínas o valor para os dançarinos foi de 18 ± 6,2% e de 17 ± 2,9 para as dançarinas. Sobre os lipídios os numerários ingeridos foram de 19,6 ± 7,6% e 21,7 ± 4,7% nesta ordem para meninos e meninas onde, p≥0,05. Em conclusão, os bailarinos do presente estudo, mostraram uma inadequação alimentar, não coerentes com as necessidades diárias de nutrientes, para sua modalidade de dança. ABSTRACT Nutritional conduct of adolescent dancers of both sexA balanced diet is a major factor for dancers because of their contribution to improving the body composition and physical fitness of this audience. The objective of the research was to evaluate the nutritional behavior of macro- and micronutrients in the diet of classic adolescent dancers. Nine male dancers of 16.4 ± 1.5 years old and female 14.5 ± 0.9 years old were evaluated. The fat percentage showed values of 8.8 ± 1.3% and 8.7 ± 2.2% for boys and girls in this order. The food consumption of the dancers was recorded in the semi-quantitative food frequency questionnaire. Boys rarely consumed milks and derivatives 28.8%, vegetables and fruits 70%, beverages 35.7% and products diet or light 100% where p˂0.05. Girls consumed snacks and canned once a week 66.7% and rarely drinks 84.6% and diet light products 92.9% where p˂0.05. Macronutrient intake was determined through a three-day dietary recall. The percentage of carbohydrate in the diet of the dancers was 57.8 ± 16.1%, of the dancers 59.7 ± 5.5%. For the protein the value for the dancers was 18 ± 6.2% and 17 ± 2.9 for the dancers. Considering lipids, the ingested numbers were 19.6 ± 7.6% and 21.7 ± 4.7% in this order for boys and girls where, p≥0.05. In conclusion, the dancers of the present study showed a feeding inadequacy, not consistent with the daily nutrient requirements for their dance modality

Assessing the reliability of retrospective reports of adverse childhood experiences among adolescents with documented childhood maltreatment

The literature suggests that childhood maltreatment is related to a higher probability of developing psychopathology and disease in adulthood. However, some authors have questioned the reliability of self-reports of maltreatment, suggesting that psychopathology at the time of evaluation affects self-reports. We evaluated the reliability of the self-reports of 79 young adults who were identified in childhood by Child Protective Services by comparing two moments of evaluation. Psychological and physical symptoms were tested to evaluate their interference with the reports. We found good to excellent agreement, with no significant correlation between the changes in self-reported experiences and the changes in physical and psychological symptoms, suggesting that the reliability of reports is not related to the health state at the time of the report