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1.
J Cell Physiol ; 234(8): 13659-13679, 2019 08.
Artículo en Inglés | MEDLINE | ID: mdl-30637720

RESUMEN

Osteosarcomas are bone tumors that frequently metastasize to the lung. Aberrant expression of the transcription factor, runt-related transcription factor 2 (RUNX2), is a key pathological feature in osteosarcoma and associated with loss of p53 and miR-34 expression. Elevated RUNX2 may transcriptionally activate genes mediating tumor progression and metastasis, including the RUNX2 target gene osteopontin (OPN/SPP1). This gene encodes a secreted matricellular protein produced by osteoblasts to regulate bone matrix remodeling and tissue calcification. Here we investigated whether and how the RUNX2/OPN axis regulates lung metastasis of osteosarcoma. Importantly, RUNX2 depletion attenuates lung metastasis of osteosarcoma cells in vivo. Using next-generation RNA-sequencing, protein-based assays, as well as the loss- and gain-of-function approaches in selected osteosarcoma cell lines, we show that osteopontin messenger RNA levels closely correlate with RUNX2 expression and that RUNX2 controls the levels of secreted osteopontin. Elevated osteopontin levels promote heterotypic cell-cell adhesion of osteosarcoma cells to human pulmonary microvascular endothelial cells, but not in the presence of neutralizing antibodies. Collectively, these findings indicate that the RUNX2/OPN axis regulates the ability of osteosarcoma cells to attach to pulmonary endothelial cells as a key step in metastasis of osteosarcoma cells to the lung.


Asunto(s)
Neoplasias Óseas/metabolismo , Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , Regulación Neoplásica de la Expresión Génica/fisiología , Invasividad Neoplásica , Osteopontina/metabolismo , Osteosarcoma/metabolismo , Animales , Neoplasias Óseas/patología , Adhesión Celular/fisiología , Línea Celular Tumoral , Células Endoteliales/metabolismo , Xenoinjertos , Humanos , Pulmón/metabolismo , Pulmón/patología , Neoplasias Pulmonares/secundario , Ratones , Ratones Endogámicos NOD , Ratones SCID , Osteosarcoma/secundario
2.
J Cell Biochem ; 119(10): 8204-8219, 2018 11.
Artículo en Inglés | MEDLINE | ID: mdl-29923217

RESUMEN

Osteoblast differentiation is controlled by transcription factor RUNX2 which temporally activates or represses several bone-related genes, including those encoding extracellular matrix proteins or factors that control cell-cell, and cell-matrix interactions. Cell-cell communication in the many skeletal pericellular micro-niches is critical for bone development and involves paracrine secretion of growth factors and morphogens. This paracrine signaling is in part regulated by "A Disintegrin And Metalloproteinase" (ADAM) proteins. These cell membrane-associated metalloproteinases support proteolytic release ("shedding") of protein ectodomains residing at the cell surface. We analyzed microarray and RNA-sequencing data for Adam genes and show that Adam17, Adam10, and Adam9 are stimulated during BMP2 mediated induction of osteogenic differentiation and are robustly expressed in human osteoblastic cells. ADAM17, which was initially identified as a tumor necrosis factor alpha (TNFα) converting enzyme also called (TACE), regulates TNFα-signaling pathway, which inhibits osteoblast differentiation. We demonstrate that Adam17 expression is suppressed by RUNX2 during osteoblast differentiation through the proximal Adam17 promoter region (-0.4 kb) containing two functional RUNX2 binding motifs. Adam17 downregulation during osteoblast differentiation is paralleled by increased RUNX2 expression, cytoplasmic-nuclear translocation and enhanced binding to the Adam17 proximal promoter. Forced expression of Adam17 reduces Runx2 and Alpl expression, indicating that Adam17 may negatively modulate osteoblast differentiation. These findings suggest a novel regulatory mechanism involving a reciprocal Runx2-Adam17 negative feedback loop to regulate progression through osteoblast differentiation. Our results suggest that RUNX2 may control paracrine signaling through regulation of ectodomain shedding at the cell surface of osteoblasts by directly suppressing Adam17 expression.


Asunto(s)
Proteína ADAM17/genética , Proteína Morfogenética Ósea 2/genética , Subunidad alfa 1 del Factor de Unión al Sitio Principal/genética , Retroalimentación Fisiológica , Osteoblastos/metabolismo , Osteogénesis/genética , Proteínas ADAM/genética , Proteínas ADAM/metabolismo , Proteína ADAM10/genética , Proteína ADAM10/metabolismo , Proteína ADAM17/metabolismo , Fosfatasa Alcalina/genética , Fosfatasa Alcalina/metabolismo , Secretasas de la Proteína Precursora del Amiloide/genética , Secretasas de la Proteína Precursora del Amiloide/metabolismo , Animales , Sitios de Unión , Proteína Morfogenética Ósea 2/metabolismo , Diferenciación Celular , Línea Celular , Línea Celular Tumoral , Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Humanos , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Ratones , Osteoblastos/citología , Comunicación Paracrina/genética , Regiones Promotoras Genéticas , Unión Proteica , Ratas , Transducción de Señal , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismo
3.
J Cell Biochem ; 118(2): 351-360, 2017 02.
Artículo en Inglés | MEDLINE | ID: mdl-27356893

RESUMEN

Osteosarcomas are the most prevalent bone tumors in pediatric patients, but can also occur later in life. Bone tumors have the potential to metastasize to lung and occasionally other vital organs. To understand how osteosarcoma cells interact with their micro-environment to support bone tumor progression and metastasis, we analyzed secreted proteins and exosomes from three human osteosarcoma cell lines. Exosome isolation was validated by transmission electron microscopy (TEM) and immuno-blotting for characteristic biomarkers (CD63, CD9, and CD81). Exosomal and soluble proteins (less than 100 kDa) were identified by mass spectrometry analysis using nanoLC-MS/MS and classified by functional gene ontology clustering. We identified a secretome set of >3,000 proteins for both fractions, and detected proteins that are either common or unique among the three osteosarcoma cell lines. Protein ontology comparison of proteomes from exosomes and exosome-free fractions revealed differences in the enrichment of functional categories associated with different biological processes, including those related to tumor progression (i.e., angiogenesis, cell adhesion, and cell migration). The secretome characteristics of osteosarcoma cells are consistent with the pathological properties of tumor cells with metastatic potential. J. Cell. Biochem. 118: 351-360, 2017. © 2016 Wiley Periodicals, Inc.


Asunto(s)
Antígenos CD/metabolismo , Biomarcadores de Tumor/metabolismo , Neoplasias Óseas/metabolismo , Exosomas/metabolismo , Proteínas de Neoplasias/metabolismo , Osteosarcoma/metabolismo , Línea Celular Tumoral , Humanos
4.
J Cell Biochem ; 118(11): 3662-3674, 2017 11.
Artículo en Inglés | MEDLINE | ID: mdl-28370561

RESUMEN

Osteosarcoma is the most common malignant bone tumor in children and adolescents. Metastasis and poor responsiveness to chemotherapy in osteosarcoma correlates with over-expression of the runt-related transcription factor RUNX2, which normally plays a key role in osteogenic lineage commitment, osteoblast differentiation, and bone formation. Furthermore, WNT/ß-catenin signaling is over-activated in osteosarcoma and promotes tumor progression. Importantly, the WNT/ß-catenin pathway normally activates RUNX2 gene expression during osteogenic lineage commitment. Therefore, we examined whether the WNT/ß-catenin pathway controls the tumor-related elevation of RUNX2 expression in osteosarcoma. We analyzed protein levels and nuclear localization of ß-catenin and RUNX2 in a panel of human osteosarcoma cell lines (SAOS, MG63, U2OS, HOS, G292, and 143B). In all six cell lines, ß-catenin and RUNX2 are expressed to different degrees and localized in the nucleus and/or cytoplasm. SAOS cells have the highest levels of RUNX2 protein that is localized in the nucleus, while MG63 cells have the lowest RUNX2 levels which is mostly localized in the cytoplasm. Levels of ß-catenin and RUNX2 protein are enhanced in HOS, G292, and 143B cells after treatment with the GSK3ß inhibitor SB216763. Furthermore, small interfering RNA (siRNA)-mediated depletion of ß-catenin inhibits RUNX2 expression in G292 cells. Thus, WNT/ß-catenin activation is required for RUNX2 expression in at least some osteosarcoma cell types, where RUNX2 is known to promote expression of metastasis related genes. J. Cell. Biochem. 118: 3662-3674, 2017. © 2017 Wiley Periodicals, Inc.


Asunto(s)
Neoplasias Óseas/metabolismo , Subunidad alfa 1 del Factor de Unión al Sitio Principal/biosíntesis , Proteínas de Neoplasias/biosíntesis , Osteosarcoma/metabolismo , Vía de Señalización Wnt , Neoplasias Óseas/genética , Neoplasias Óseas/patología , Línea Celular Tumoral , Subunidad alfa 1 del Factor de Unión al Sitio Principal/genética , Regulación Neoplásica de la Expresión Génica , Humanos , Metástasis de la Neoplasia , Proteínas de Neoplasias/genética , Osteosarcoma/genética , Osteosarcoma/patología
5.
Biol Res ; 49(1): 28, 2016 Jun 03.
Artículo en Inglés | MEDLINE | ID: mdl-27255980

RESUMEN

BACKGROUND: Tear desiccation on a glass surface followed by transmitted-light microscopy has served as diagnostic test for dry eye. Four distinctive morphological domains (zones I, II, III and transition band) have been recently recognized in tear microdesiccates. Physicochemical dissimilarities among those domains hamper comprehensive microscopic examination of tear microdesiccates. Optimal observation conditions of entire tear microdesiccates are now investigated. One-µl aliquots of tear collected from individual healthy eyes were dried at ambient conditions on microscope slides. Tear microdesiccates were examined by combining low-magnification objective lenses with transmitted-light microscopy (brightfield, phase contrasts Ph1,2,3 and darkfield). RESULTS: Fern-like structures (zones II and III) were visible with all illumination methods excepting brightfield. Zone I was the microdesiccate domain displaying the most noticeable illumination-dependent variations, namely transparent band delimited by an outer rim (Ph1, Ph2), homogeneous compactly built structure (brightfield) or invisible domain (darkfield, Ph3). Intermediate positions of the condenser (BF/Ph1, Ph1/Ph2) showed a structured roughly cylindrical zone I. The transition band also varied from invisibility (brightfield) to a well-defined domain comprising interwoven filamentous elements (phase contrasts, darkfield). CONCLUSIONS: Imaging of entire tear microdesiccates by transmitted-light microscopy depends upon illumination. A more comprehensive description of tear microdesiccates can be achieved by combining illumination methods.


Asunto(s)
Desecación/métodos , Síndromes de Ojo Seco/diagnóstico por imagen , Microscopía de Contraste de Fase/métodos , Lágrimas/diagnóstico por imagen , Adolescente , Adulto , Femenino , Humanos , Luz , Iluminación , Masculino , Valores de Referencia , Reproducibilidad de los Resultados , Propiedades de Superficie , Lágrimas/metabolismo , Adulto Joven
6.
Food Technol Biotechnol ; 54(4): 462-467, 2016 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-28115904

RESUMEN

Fruits and vegetables are considered a good source of polyphenols and antioxidant capacities which are beneficial in protecting the human body against damage induced by reactive species. The objective of this work is to conduct an assessment of the polyphenol content and antioxidant activities of different fruit (kiwi, pear, green apple, raspberry, blackberry, strawberry and blueberry) and vegetable (pumpkin, green and red pepper) extracts using both chemical extraction and a modified in vitro digestive enzymatic extraction in order to compare results. Polyphenol content and antioxidant capacity of different fruits, vegetables and fruit juices were determined by Folin-Ciocalteu and FRAP methods, respectively. It was observed that polyphenol content expressed as gallic acid equivalents of extracts obtained with the two extraction methods was significantly (p<0.05) different (on average 310.3 and 231.8 mg per 100 g of fresh sample in enzymatic and methanolic extracts, respectively). Antioxidant capacity was also significantly (p<0.05) different in the extracts obtained by the two methods, with higher values in enzymatic extracts (1.91 mmol of Fe2+ per 100 g of fresh sample). Analyses of apple samples with and without skin also revealed important differences related to methodology and composition. Additionally, the original enzymatic extraction method was improved to avoid interferences caused by the presence of protein residues in the extract.

7.
Biol Res ; 47: 25, 2014 Jun 04.
Artículo en Inglés | MEDLINE | ID: mdl-25026954

RESUMEN

BACKGROUND: Fern-like crystalloids form when a microvolume of tear is allowed to dry out at ambient conditions on a glass surface. Presence of crystalloids in tear "microdesiccates" is used to evaluate patients with Dry-Eye disease. This study aims to examine morphologically the desiccation process of normal tear fluid and to identify changes associated with accelerated tear evaporation. Tear microdesiccates from healthy (Non-Dry Eye) and Dry Eye subjects were produced at ambient conditions. Microdesiccate formation was monitored continuously by dark-field video microscopy. Additionally, accelerated desiccation of tear samples from healthy subjects was conducted under controlled experimental conditions. Particular morphological domains of tear microdesiccates and their progressive appearance during desiccation were compared. RESULTS: In normal tear microdesiccates, four distinctive morphological domains (zones I, II, III and transition band) were recognized. Stepwise formation of those domains is now described. Experimentally accelerated desiccation resulted in marked changes in some of those zones, particularly involving either disappearance or size reduction of fern-like crystalloids of zones II and III. Tear microdesiccates from Dry Eye subjects may also display those differences and be the expression of a more synchronous formation of microdesiccate domains. CONCLUSION: Morphological characteristics of tear microdesiccates can provide insights into the relative rate of tear evaporation.


Asunto(s)
Desecación , Vidrio , Lágrimas/química , Adulto , Cristalización , Síndromes de Ojo Seco/diagnóstico , Humanos , Hidrodinámica , Microscopía por Video , Persona de Mediana Edad
8.
Biol Res ; 46(3): 299-305, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-24346079

RESUMEN

Desiccation of human tears on glass surfaces results in fern-like crystalloids. This phenomenon has been associated with tear normality (Tear Ferning Test, TFT) and is used as a diagnostic aid to evaluate patients with Dry-Eye disease. However, TFT is focused on the assessment of only a minor fraction of desiccated tear samples and considers only the relative abundance and density of fern-like crystalloids. The aim of this study was to characterize morphologically entire desiccated micro volumes of tears from healthy donors. Tear samples were collected from 23 healthy young adult volunteers. Tear aliquots (1-3 µL) were allowed to dry on glass surfaces under ambient conditions of temperature (15-25°C) and relative humidity (40-45%). Dry samples were analyzed by dark-field microscopy. Morphometric data were acquired with Image J software. Tear volume was positively correlated with both area and time of desiccation. Morphological features of multiple microdesiccates produced from a single subject displayed striking similarities whereas tear microdesiccates from different healthy subjects displayed consistent differences but shared a common general design. This design may be mostly represented by the occurrence of four distinctive zones, named as zones I, II, III and Transition band. The main features of these zones are described.


Asunto(s)
Cristalización , Lágrimas/química , Adolescente , Desecación/métodos , Femenino , Humanos , Masculino , Adulto Joven
9.
Cogn Neurodyn ; 13(4): 409-416, 2019 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-31354885

RESUMEN

In this work we present a group of theoretical models for reaction times arising from simple-choice task tests. In particular, we argue for the inclusion of a shifted version of the Gamma distribution as a theoretical model based on a mathematical result on first hitting times. We contrast the goodness-of-fit of those models with the Ex-Gaussian distribution, using data from recently published experiments. The evidence of the results obtained highlights the convenience of proposing theoretical models for reaction times instead of models acting exclusively as quantitative distribution measurements.

10.
Gene ; 710: 246-257, 2019 Aug 20.
Artículo en Inglés | MEDLINE | ID: mdl-31176732

RESUMEN

Osteosarcoma is the most common primary bone tumor during childhood and adolescence. Several reports have presented data on serum biomarkers for osteosarcoma, but few reports have analyzed circulating microRNAs (miRNAs). In this study, we used next generation miRNA sequencing to examine miRNAs isolated from microvesicle-depleted extracellular vesicles (EVs) derived from six different human osteosarcoma or osteoblastic cell lines with different degrees of metastatic potential (i.e., SAOS2, MG63, HOS, 143B, U2OS and hFOB1.19). EVs from each cell line contain on average ~300 miRNAs, and ~70 of these miRNAs are present at very high levels (i.e., >1000 reads per million). The most prominent miRNAs are miR-21-5p, miR-143-3p, miR-148a-3p and 181a-5p, which are enriched between 3 and 100 fold and relatively abundant in EVs derived from metastatic SAOS2 cells compared to non-metastatic MG63 cells. Gene ontology analysis of predicted targets reveals that miRNAs present in EVs may regulate the metastatic potential of osteosarcoma cell lines by potentially inhibiting a network of genes (e.g., MAPK1, NRAS, FRS2, PRCKE, BCL2 and QKI) involved in apoptosis and/or cell adhesion. Our data indicate that osteosarcoma cell lines may selectively package miRNAs as molecular cargo of EVs that could function as paracrine agents to modulate the tumor micro-environment.


Asunto(s)
Neoplasias Óseas/genética , Vesículas Extracelulares/genética , Secuenciación de Nucleótidos de Alto Rendimiento/métodos , MicroARNs/genética , Osteosarcoma/genética , Apoptosis , Adhesión Celular , Línea Celular Tumoral , Redes Reguladoras de Genes , Humanos , Metástasis de la Neoplasia , Análisis de Secuencia de ARN/métodos
11.
Cornea ; 36(2): 229-235, 2017 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-28060072

RESUMEN

PURPOSE: Morphological features of tear microdesiccates on glass surfaces have been associated with tear fluid status. Tear-film lipids play a critical role in the pathophysiology of some ocular surface disorders. Tear microdesiccates display 4 distinctive morphological domains (zones I, II, III, and transition band). In this study, we investigated the lipid location in tear microdesiccates. METHODS: Tear from individual healthy eyes (assessed by symptoms, signs, and slit-lamp examination) was collected using absorbing minisponges. One-µL aliquots were allowed to dry under ambient conditions on microscope slides. Tear microdesiccates were examined by various transmitted light microscopy methods. Tear lipids were located both by partition experiments using 2 lipophilic dyes (Oil red O and Nile blue A) mixed with tear fluid under conditions preserving morphological features of microdesiccates and by assessing the effect of 2 solvents markedly differing in polarity (water and ethanol) on the morphology of particular domains of preformed microdesiccates. RESULTS: During desiccation, both Nile blue A and Oil red O became preferentially located in the outermost domain of tear microdesiccates (zone I) without affecting the formation of major fern-like crystalloids (zones II and III). Low volumes of water drastically affected fern-like crystalloids, whereas the gross morphology of zone I was maintained. Contrarily, ethanol, a less polar solvent, was a fixative for fern-like crystalloids, although it markedly affected the bulk of zone I by extracting liquid droplets out of microdesiccates and visibilizing some filamentous subcomponents. CONCLUSIONS: Zone I is a hydrophobic domain, whereas zones II and III are highly hydrophilic domains of tear microdesiccates. Zone I represents a lipid-rich structure.


Asunto(s)
Lípidos/análisis , Lágrimas/química , Adolescente , Compuestos Azo/farmacología , Colorantes/farmacología , Desecación , Femenino , Voluntarios Sanos , Humanos , Masculino , Oxazinas/farmacología , Lágrimas/efectos de los fármacos , Adulto Joven
12.
Arch Latinoam Nutr ; 56(4): 361-5, 2006 Dec.
Artículo en Español | MEDLINE | ID: mdl-17425182

RESUMEN

The high prevalence of non transmissible chronic diseases (NCD) related to food consumption had increased the studies conducted to investigate the relationship between diet and health. A smaller incidence of NCD, with food patterns with high consumption of fruits and vegetables has been observed and chemical compounds of these foods have been one of the main subjects of the actual research in the reaqltion between food consumption and health. The effect of vegetable foods has been attributed to various nutrients and bioactive compounds with antioxidant activity. In order to determine the antioxidant capacity of vegetable foods cultivated in Chile, natural fruits and vegetables were analyzed according to the FRAP (ferric reducing activity power) method, reading to the 4 minutes. In vegetables, the values were between 0.002 and 1.91 milimoles of Fe/l00 g for cooked carrot and red pepper respectively. The values of the fruits ranged between 0.02 milimoles of Fe/100 g for the cucumber and 12.32 for maqui, the berries studies showed values between 3.10 for strawberry and 3.55 for wild blackberry. Lemmon and quince with 0.25 and 0.23 respectively are located in the intermediate level and the lowest values within the fruits corresponded to apple (fuji variety) and peaches.


Asunto(s)
Antioxidantes/análisis , Flavonoides/análisis , Frutas/química , Fenoles/análisis , Verduras/química , Chile , Conducta Alimentaria , Conservación de Alimentos , Humanos , Oxidación-Reducción , Polifenoles
13.
Appl Physiol Nutr Metab ; 37(5): 997-1002, 2012 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-22871100

RESUMEN

The aim of this study was to assess red blood cell glutathione from insulin-sensitive and insulin-resistant individuals before and after an oral glucose dose. Fifteen healthy, young (24 ± 5 years), nonobese (23 ± 2 kg·m⁻²), insulin-sensitive (ISI composite = 6.0 ± 1.2) individuals and 14 healthy, young (22 ± 2 years), nonobese (24 ± 2 kg·m⁻²), insulin-resistant (ISI composite = 2.7 ± 1.1) individuals received a 75 g oral glucose dose. Blood samples were drawn before and for 2 h after glucose ingestion for red blood cell glutathione and serum glucose and insulin concentrations. Glycemia before and after glucose ingestion was similar between groups (p = 0.17), which suggest that hyperinsulinemia compensated impaired insulin sensitivity. Red blood cell total (p = 0.81), reduced (p = 0.79), and oxidized (p = 0.88) glutathione concentrations were similar between groups under fasting and postprandial conditions. However, in response to glucose, increases in total and reduced glutathione concentrations were found at the end of the 2 h assessment period in both groups (p < 0.05). Direct associations between postprandial glucose response and red blood cell total (r = 0.52; p < 0.05) and oxidized (r = 0.61; p = 0.02) glutathione concentrations were observed only in insulin-sensitive subjects. In conclusion, healthy individuals differing in their degree of insulin resistance showed similar red blood cell glutathione concentrations under non-glucose- and glucose-stimulated conditions.


Asunto(s)
Eritrocitos/metabolismo , Glutatión/sangre , Resistencia a la Insulina , Adulto , Glucemia/análisis , Femenino , Prueba de Tolerancia a la Glucosa , Glutatión/metabolismo , Humanos , Hiperglucemia/sangre , Hiperglucemia/fisiopatología , Insulina/sangre , Masculino , Oxidación-Reducción , Periodo Posprandial , Índice de Severidad de la Enfermedad , Adulto Joven
14.
Biol. Res ; 49: 1-10, 2016. ilus, tab
Artículo en Inglés | LILACS | ID: biblio-950855

RESUMEN

BACKGROUND: Tear desiccation on a glass surface followed by transmitted-light microscopy has served as diagnostic test for dry eye. Four distinctive morphological domains (zones I, II, III and transition band) have been recently recognized in tear microdesiccates. Physicochemical dissimilarities among those domains hamper comprehensive microscopic examination of tear microdesiccates. Optimal observation conditions of entire tear microdesiccates are now investigated. One-µl aliquots of tear collected from individual healthy eyes were dried at ambient conditions on microscope slides. Tear microdesiccates were examined by combining low-magnification objective lenses with transmitted-light microscopy (brightfield, phase contrasts Ph1,2,3 and darkfield. RESULTS: Fern-like structures (zones II and III) were visible with all illumination methods excepting brightfield. Zone I was the microdesiccate domain displaying the most noticeable illumination-dependent variations, namely transparent band delimited by an outer rim (Ph1, Ph2), homogeneous compactly built structure (brightfield) or invisible domain (darkfield, Ph3). Intermediate positions of the condenser (BF/Ph1, Ph1/Ph2) showed a structured roughly cylindrical zone I. The transition band also varied from invisibility (brightfield) to a well-defined domain comprising interwoven filamentous elements (phase contrasts, darkfield. CONCLUSIONS: Imaging of entire tear microdesiccates by transmitted-light microscopy depends upon illumination. A more comprehensive description of tear microdesiccates can be achieved by combining illumination methods.


Asunto(s)
Humanos , Masculino , Femenino , Adolescente , Adulto , Adulto Joven , Lágrimas/diagnóstico por imagen , Síndromes de Ojo Seco/diagnóstico por imagen , Microscopía de Contraste de Fase/métodos , Desecación/métodos , Valores de Referencia , Propiedades de Superficie , Lágrimas/metabolismo , Iluminación , Reproducibilidad de los Resultados , Luz
15.
Biol. Res ; 47: 1-10, 2014. ilus, graf
Artículo en Inglés | LILACS | ID: biblio-950721

RESUMEN

BACKGROUND: Fern-like crystalloids form when a microvolume of tear is allowed to dry out at ambient conditions on a glass surface. Presence of crystalloids in tear "microdesiccates" is used to evaluate patients with Dry-Eye disease. This study aims to examine morphologically the desiccation process of normal tear fluid and to identify changes associated with accelerated tear evaporation. Tear microdesiccates from healthy (Non-Dry Eye) and Dry Eye subjects were produced at ambient conditions. Microdesiccate formation was monitored continuously by dark-field video microscopy. Additionally, accelerated desiccation of tear samples from healthy subjects was conducted under controlled experimental conditions. Particular morphological domains of tear microdesiccates and their progressive appearance during desiccation were compared. RESULTS: In normal tear microdesiccates, four distinctive morphological domains (zones I, II, III and transition band) were recognized. Stepwise formation of those domains is now described. Experimentally accelerated desiccation resulted in marked changes in some of those zones, particularly involving either disappearance or size reduction of fern-like crystalloids of zones II and III. Tear microdesiccates from Dry Eye subjects may also display those differences and be the expression of a more synchronous formation of microdesiccate domains. CONCLUSION: Morphological characteristics of tear microdesiccates can provide insights into the relative rate of tear evaporation.


Asunto(s)
Humanos , Adulto , Persona de Mediana Edad , Lágrimas/química , Desecación , Vidrio , Síndromes de Ojo Seco/diagnóstico , Microscopía por Video , Cristalización , Hidrodinámica
16.
Biol. Res ; 46(3): 299-305, 2013. ilus, graf, tab
Artículo en Inglés | LILACS | ID: lil-692198

RESUMEN

Desiccation of human tears on glass surfaces results in fern-like crystalloids. This phenomenon has been associated with tear normality (Tear Ferning Test, TFT) and is used as a diagnostic aid to evaluate patients with Dry-Eye disease. However, TFT is focused on the assessment of only a minor fraction of desiccated tear samples and considers only the relative abundance and density of fern-like crystalloids. The aim of this study was to characterize morphologically entire desiccated micro volumes of tears from healthy donors. Tear samples were collected from 23 healthy young adult volunteers. Tear aliquots (1-3 μL) were allowed to dry on glass surfaces under ambient conditions of temperature (15-25°C) and relative humidity (40-45%). Dry samples were analyzed by dark-field microscopy. Morphometric data were acquired with Image J software. Tear volume was positively correlated with both area and time of desiccation. Morphological features of multiple microdesiccates produced from a single subject displayed striking similarities whereas tear microdesiccates from different healthy subjects displayed consistent differences but shared a common general design. This design may be mostly represented by the occurrence of four distinctive zones, named as zones I, II, III and Transition band. The main features of these zones are described.


Asunto(s)
Adolescente , Femenino , Humanos , Masculino , Adulto Joven , Cristalización , Lágrimas/química , Desecación/métodos
17.
Int J Food Sci Nutr ; 54(2): 119-26, 2003 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-12701368

RESUMEN

A study was performed to examine the rate of digestion of available carbohydrate in legumes and its mixtures with cereals, prepared as commonly eaten. The legumes and cereals studied were lentil (Lens sculenta), pea (Pisum sativum), bean (Phaseolus vulgaris, var tortola), rice (Oryza sativa) and spaghetti. Foods were purchased at the city market. Total starch content and the carbohydrate digestion rates were determined using the enzymatic method proposed by Englyst et al. Total starch levels ranged from 7.78 g/100 g in cooked flour bean to 20.6 g/100 g in a bean-spaghetti dish, and dietary fiber contents ranged from 2.4 g/100 g in a cooked 70:30 lentil-rice mixture to 5.26 g/100 g in a cooked whole bean. The rapid digestion rate carbohydrates showed values from 4.8 in the bean soup to 8.9 in the bean-spaghetti combination. The same results show, expressed as rapid available glucose (RAG), the amount of rapid carbohydrate/100 g food or meal as eaten, and as the starch digestion index (SDI), the percentage of rapid carbohydrate digestion rate in relation to the total amount of carbohydrate. The RAG values ranged between 5.0 for cooked beans and 10 for cooked beans and spaghetti, and the SDI ranged between 40 for cooked pea flour and 62 for cooked bean flour. Legumes prepared as soup showed a higher rapid digestion rate than legumes prepared as whole grain. The bean-spaghetti based-meal and the lentil-based meal showed glycemic index mean and standard deviation values of 76.8 +/- 43.4 and 49.3 +/- 29.5, RAG values of 7.0 and 6.0, and SDI values of 57 and 54, respectively. The knowledge of the importance of the carbohydrate digestion rates in human health in increasing, and probably will soon be used in the development of the food pyramid. The foods with a moderate fraction of rapid digestion rate, such as legumes, should be included in the base of the pyramid.


Asunto(s)
Carbohidratos de la Dieta/metabolismo , Digestión/fisiología , Fabaceae/química , Adulto , Glucemia/análisis , Grano Comestible , Manipulación de Alimentos , Índice Glucémico , Humanos , Masculino , Almidón
18.
Nutrire Rev. Soc. Bras. Aliment. Nutr ; 24: 71-83, dez. 2002. tab
Artículo en Portugués | LILACS | ID: lil-356354

RESUMEN

O EDTA é utilizado como aditivo em diferentes alimentos. Atualmente não existe informação sobre o consumo de EDTA em populações de diferentes países da América Latina. O objetivo do presente estudo foi avaliar de forma descritiva o consumo de EDTA em escolares brasileiros, chilenos e mexicanos. Um total de 677 crianças (355 brasileiros, 204 mexicanos e 118 chilenos) entre 6 e 13 anos de idade, de escolas privadas, participaram do estudo. A ingestão do aditivo foi avaliada no Brasil e no Chile através de um recordatório de 24 horas e no México utilizando um questionário de freqüência alimentar. No Brasil, a principal fonte de EDTA foi a margarina (30 por cento), seguida da maionese (24 por cento) e molhos para saladas (23 por cento)...


Asunto(s)
Humanos , Masculino , Femenino , Niño , Adolescente , Nutrición del Niño , Ácido Edético , Aditivos Alimentarios , Ingestión de Alimentos
19.
Rev. chil. nutr ; 15(2): 89-92, ago. 1987. tab
Artículo en Español | LILACS | ID: lil-55035

RESUMEN

Se presenta un análisis acerca de los factores dietéticos que influyen sobre los valores de la relación proteína-energía dietética (P%) y en las formas más racionales de su aplicación. El análisis realizado en el presente trabajo permite concluir que para lograr una aplicación e interpretación adecuada del P%: se deben ajustar sus valores a un valor de la relación G/P de 2,5 de necesario considerar la ingesta energética de los individuos que consumen la dieta, y el P% no se debe aplicar a la evaluación nutricional de las preparaciones o dietas tal como se consumen


Asunto(s)
Humanos , Encuestas sobre Dietas , Necesidad Energética , Proteínas en la Dieta/análisis , Guatemala
20.
Arch. latinoam. nutr ; 46(1): 42-6, mar. 1996. tab
Artículo en Español | LILACS | ID: lil-213177

RESUMEN

La necesidad de un incremento de la ingesta de fibra dietética (FD) justifica el estudio de las algas como una fuente importante de ella. El objetivo del trabajo fue estudiar en ratas, el efecto de la FD de las algas, sobre el peso de las deposiciones y digestibilidad aparente de energía, proteínas y fibra. Las algas estudiadas fueron el cochayuyo (fronda) y ulte (parte basal) (Durvilleae antarctica), luche verde (Ulva lactuca) y luche rojo, (Porphyra columbina) cocidos e ebullición y secados a 55ºC bajo corriente de aire. Ratas machos Wistar de 25 días de edad fueron alimentadas durante un período experimental de 29 días con dietas de caseína con inclusión de 10 por ciento de FD de las algas. Como grupo control se utilizó una dieta sin fibra. La FD de las algas varió entre 58,2 g/100 g a 75,6 g/100 (peso seco), del cual 37,9 por ciento a 52,4 por ciento fueron soluble. En relación al control: a) Se encontró disminución significativa de la digestibilidad de proteínas y deposiciones. La digestibilidad de la FD varió entre 21.1 por ciento a 43.1 por ciento. Se demuestra que las algas constituyen una excelente fuente de FD, que puede ser beneficiosa para prevenir o tratar enfermedades derivadas de su carencia


Asunto(s)
Ratas , Chlorophyta , Fibras de la Dieta/estadística & datos numéricos , Rhodophyta , Algas Marinas
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