RESUMO
Objective: To investigate the effects of RCCS simulated microgravity on the metabolism of the human keratinocyte cell line HaCaT. Methods: The rotary cell culture system (RCCS) was used to simulate the microgravity environment, and HaCaT cells were cultured in vitro and divided randomly into simulated microgravity group (SMG) and normal gravity group (NG). The two group HaCaT cells were collected respectively after 1 d, 2 d and 3 d culture, and the samples were analyzed by LC/MS metabolomics. The differential metabolites between the SMG and NG cells were identified with partial least squares discriminant analysis ((O)PLS-DA), and the data were input into the KEGG database for the construction and functional analysis of metabolic pathways. Results: Comparing to NG cells, after 1 d culture, there were 74 different metabolites in SMG cells, among which 16 were up-regulated and 58 were down-regulated; after 2 d culture, there were 89 different metabolites, among which 15 were up-regulated and 74 down-regulated; after 3d culture, there were 100 different metabolites, of which 23 were up-regulated and 77 were down-regulated. The differentially expressed 49 metabolites (VIP>1 and P<0.05) after 3 d were set as target metabolites, within which the sphingosine, glutamate, and docosapentaenoic acid were down-regulated, and dehydrated sorbitol was up-regulated. KEGG analysis indicated that the metabolic pathways involved were amino acid metabolism, lipid metabolism, cell proliferation and apoptosis, substance transport, catabolism, and signal transduction. Conclusion: RCCS simulated microgravity may have significant impacts on keratinocyte metabolism mainly involving metabolites such as sphingolipids and glutamate as well as the related signaling pathways.
RESUMO
A method of high performance liquid chromatography coupled with time-of-flight mass spectrometry was used to detect the endogenous metabolites changes in plasma of normal rats, rats of dampness stagnancy due to spleen deficiency and Astragalus flavone component intervention rats. Metabolism map of rat plasma was obtained and the mechanism of Astragalus flavonoids on dampness stagnancy due to spleen deficiency was studied. Rat model with dampness stagnancy due to spleen deficiency was established by high fat and low protein diet plus load swimming. Liquid chromatography tandem mass spectrometry was used for the analysis of rat plasma sample, and 0.05％ formic acid water with 0.05％ formic acid acetonitrile as the mobile phase was applied in gradient elution with Halo C18 chromatographic column. In this study, partial least squares discriminant analysis and variance analysis were used to screen the potential biomarkers, it was found that the metabolic profile of the Astragalus flavonoids was different from that of the model group, which was close to that of the normal group. A total of 11 potential biomarkers were identified, including glycerol phospholipids, sphingolipids, amino acids, and so on. The metabolic pathways of biomarkers including three tricarboxylic acid cycle, glycerol phospholipid metabolism, fatty acid metabolism, amino acid metabolism and so on, which mainly related to energy metabolism and fat metabolism in the body. Related indexes of rats with syndrome of spleen deficiency of water and dampness were significantly callback after Astragalus flavone intervention, including macro indicators such as body weight, independent activities and micro indicators such as metabolic markers, blood lipids and others. The result showed that Astragalus flavonoids played the role of strengthening the spleen and draining the water mainly through regulating the energy metabolism, fat metabolism and so on.
RESUMO
La enfermedad de Pompe (EP), también conocida como déficit de maltasa ácida y glucogenosis tipo II, es una enfermedad por depósito de glucógeno consecuencia del déficit de la enzima alfa glicosidasa ácida lisosomal (GAA), no obstante, alteraciones en los mecanismos de autofagia también determinan la patogenia de la enfermedad. Sus manifestaciones son principalmente a nivel del músculo esquelético y cardíaco aunque también hay compromiso extramuscular. La misma es de evolución progresiva y con un patrón de herencia autosómico recesivo. Existen dos formas clínicas principales que son la EP infantil, con o sin miocardiopatía y la forma de inicio tardío. El diagnóstico se realiza demostrando la actividad descendida de la GAA con la técnica de la gota de sangre seca, complementando con un segundo análisis de la actividad enzimática o análisis genético. El tratamiento disponible actualmente es la terapia de remplazo enzimático con GAA recombinante humana (rhGAA). Se describe el primer caso de Enfermedad de Pompe en el adulto en Uruguay confirmado genéticamente.
Pompe disease in Uruguay. The first case in an adult genetically confirmed. Pompe disease, also known as type II glycogenosis, is a progressive autosomal recesive glycogen storage disease caused by a deficiency of lysosomal acid-a-glucosidase (GAA), primarily in skeletal and cardiac muscle, and by defects in autophagy, with an age of onset ranging from infancy through adulthood. In adult onset the tipical presentation is with a limb girdle dystrophy pattern (that engaged especially the hip girdle) or dyspnea secondary to diaphragm weakness. A simple blood-based assay to measure the level of α-glucosidase activity, like dried blood spot test, is the optimal initial test and is needed a second test to confirm the disease (alpha glucosidasa activity in cultured fibroblast or muscle tissue or by genetic testing). An early diagnosis of Pompe disease will improve patient outcomes as care standards including enzyme replacement therapy can be applied and complications can be anticipated. The introduction of enzyme replacement therapy for GAA deficiency using recombinant human GAA (rhGAA) changed the course of the disease with an improvement especially in infantile forms but also in late onset Pompe disease. We describe our first case of adult onset Pompe disease in Uruguay.
RESUMO
The purpose of this study was to diagnose and characterize the leaf ergastic substance of Oenocarpus bacaba Mart., O. distichus Mart., O. mapora H. Karst. and O. minor Mart. by light and scanning electron microscopy, histochemical analyses and physical microanalyses. Transverse and longitudinal sections as well as maceration were made in fixed botanical material. The histochemical analyses were made in non-fixed botanical material, using specific tests for mucilage, starch and silica. An Energy Dispersive Spectroscopy (EDS) detector was used for the physical microanalyses. In the Oenocarpus Mart. species studied, the leaf ergastic substances corresponded to amorphous mucilage, simple-type polyhedrical starch grain and opaline silica in the shape of elliptical and spherical, globular of pointed surface bodies, both with silicon dioxide in the correct concentration. The microscopic observations, histochemical analyses and physical microanalyses permitted us to diagnose, characterize and elucidate the structure and ultrastructure of the ergastic substances occurring in the leaves of the analyzed Oenocarpus Mart. taxa.
O presente estudo teve por objetivo diagnosticar e caracterizar as substâncias ergásticas foliares de Oenocarpus bacaba Mart., O. distichus Mart., O. mapora H. Karst. e O. minor Mart. através de microscopias óptica e eletrônica de varredura, análises histoquímicas e microanálises físicas. Secções transversais e longitudinais, assim como maceração foram realizadas em material botânico fixado. As análises histoquímicas foram empregadas em material botânico in natura, seguindo-se protocolos específicos para mucilagem, amido e sílica. Microanálises físicas foram feitas com Energy Dispersive Spectroscopy (EDS) detector. Nas espécies estudadas de Oenocarpus Mart., as substâncias ergásticas foliares correspondem a mucilagem amorfa; grãos de amido poliédricos do tipo simples e sílica opalina sob a forma de corpos elípticos e esféricos-globosos de superfície espiculada, ambos com elevado teor de dióxido de silício. As observações microscópicas, os testes histoquímicos e as microanálises físicas permitiram diagnosticar, caracterizar e elucidar a estrutura e ultra-estrutura das substâncias ergásticas ocorrentes nas folhas dos táxons analisados de Oenocarpus Mart.