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The key aspect of neonatal meningitis is related to the ability of pathogens to invade the blood-brain barrier (BBB) and to penetrate the central nervous system. In the present study we show that, in an in vitro model of BBB, on the basis of co-culturing primary bovine brain endothelial cells (BBEC) and primary bovine retinal pericytes (BRPC), Escherichia coli infection determines changes of transendothelial electrical resistance (TEER) and permeability (Pe) to sodium fluorescein. In the co-culture model, within BBEC, bacteria are able to stimulate cytosolic and Ca(2+)-independent phospholipase A2 (cPLA2 and iPLA2 ) enzyme activities. In supernatants of E. coli-stimulated co-cultures, an increase in prostaglandins (PGE2) and VEGF production in comparison with untreated co-cultures were found. Incubation with E. coli in presence of AACOCF3 or BEL caused a decrease of PGE2 and VEGF release. SEM and TEM images of BBEC and BRPC showed E. coli adhesion to BBEC and BRPC but only in BBEC the invasion occurs. VEGFR-1 but not VEGFR-2 blockade by the specific antibody reduced E. coli invasion in BBEC. In our model of BBB infection, a significant loss of BRPC was observed. Following VEGFR-1, but not VEGFR-2 blockade, or in presence of AACOCF3 or BEL, elevated TEER values, reduced permeability and BRPC loss were found. These data suggest that VEGFR-1 negatively regulates BRPC survival and its blockade protects the barrier integrity. PGs and VEGF could exert a biological effect on BBB, probably by BRPC coverage ablation, thus increasing BBB permeability. Our results show the role played by the BBEC as well as BRPC during a bacterial attack on BBB. A better understanding of the mechanisms by which E. coli enter the nervous system and how bacteria alter the communication between endothelial cells and pericytes may provide exciting new insight for clinical intervention.
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Barrera Hematoencefálica/microbiología , Barrera Hematoencefálica/fisiopatología , Endotelio Vascular/patología , Infecciones por Escherichia coli/fisiopatología , Escherichia coli/patogenicidad , Pericitos/patología , Receptor 1 de Factores de Crecimiento Endotelial Vascular/fisiología , Animales , Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/farmacología , Adhesión Bacteriana/fisiología , Barrera Hematoencefálica/patología , Bovinos , Permeabilidad de la Membrana Celular/fisiología , Supervivencia Celular/fisiología , Células Cultivadas , Técnicas de Cocultivo , Dinoprostona/metabolismo , Modelos Animales de Enfermedad , Impedancia Eléctrica , Endotelio Vascular/microbiología , Escherichia coli/fisiología , Infecciones por Escherichia coli/metabolismo , Infecciones por Escherichia coli/patología , Fluoresceína , Colorantes Fluorescentes , Pericitos/microbiología , Fosfolipasas A2/metabolismo , Receptor 1 de Factores de Crecimiento Endotelial Vascular/efectos de los fármacos , Receptor 1 de Factores de Crecimiento Endotelial Vascular/inmunologíaRESUMEN
Most world countries are experiencing a remarkable aging process. Meanwhile, 50 million people are affected by Alzheimer's disease (AD) and related dementia and there is an increasing trend in the incidence of these major health problems. In order to address these, the increasing evidence suggesting the protective effect of dietary interventions against cognitive decline during aging may suggest a response to this challenge. There are nutrients with a neuroprotective effect. However, Western diets are poor in healthy n-3 polyunsaturated fatty acids (n-3 PUFAs), such as docosahexaenoic acid (DHA), iodine (I), and other nutrients that may protect against cognitive aging. Given DHA richness in chub mackerel (Scomber colias), high vitamin B9 levels in quinoa (Chenopodium quinoa), and I abundance in the seaweed Saccorhiza polyschides, a functional hamburger rich in these nutrients by using these ingredients was developed and its formulation was optimized in preliminary testing. The effects of culinary treatment (steaming, roasting, and grilling vs. raw) and digestion on bioaccessibility were evaluated. The hamburgers had high levels of n-3 PUFAs in the range of 42.0-46.4% and low levels of n-6 PUFAs (6.6-6.9%), resulting in high n-3/n-6 ratios (>6). Bioaccessibility studies showed that the hamburgers could provide the daily requirements of eicosapentaenoic acid (EPA) + DHA with 19.6 g raw, 18.6 g steamed, 18.9 g roasted, or 15.1 g grilled hamburgers. Polyphenol enrichment by the seaweed and antioxidant activity were limited. The hamburgers contained high levels of Se and I at 48-61 µg/100 g ww and 221-255 µg/100 g ww, respectively. Selenium (Se) and I bioaccessibility levels were 70-85% and 57-70%, respectively, which can be considered high levels. Nonetheless, for reaching dietary requirements, considering the influence of culinary treatment and bioaccessibility, 152.2-184.2 g would be necessary to ensure daily Se requirements and 92.0-118.1 g for I needs.
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Chub mackerel (Scomber colias) is an underutilised fish abundant in the North Atlantic and a nutritious food with high docosahexaenoic acid (DHA), selenium (Se), and vitamin B12 contents, affected by seasonality. Hence, the bioaccessibility of fatty acid (FA), Se, and vitamin B12 in chub mackerel was studied. Daily consumptions of 51 g, 35 g, and 14 g of March, June, and October chub mackerel, respectively, would ensure EPA + DHA requirements. Vitamin B12 requirements would be met by daily consumptions of 15-18 g of chub mackerel. For Se, larger amounts would be needed, 126-133 g/day. For FA, bioaccessibility often exceeded 80 %. Additionally, Se and vitamin B12 bioaccessibilities were 91-95 % and 77-83 %, respectively. Thus, bioaccessibility-based consumption frequencies for meeting dietary requirements are similar to those above, being the highest consumption frequencies associated to Se requirements, 137-146 g/day. Chub mackerel proved to be an excellent source of bioaccessible essential nutrients.
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Cyprinidae , Perciformes , Selenio , Animales , Ácidos Grasos , Vitamina B 12RESUMEN
[This corrects the article DOI: 10.3389/fnut.2022.925771.].
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Objective: Describing the availability and nutritional composition of the most commonly available street foods in Ashgabat, Turkmenistan. Methods: One hundred sixty-one street food vending sites (six public markets) were assessed, through a collection of data on vending sites' characteristics and food availability, and samples of commonly available foods (21 homemade; 11 industrial), for chemical analysis. Results: Fruit, beverages, and food other than fruit were available in 6.8, 29.2, and 91.9% of all vending sites, respectively. Regarding the latter, 52.7% of the vending sites sold only homemade products (main dishes, snacks, cakes, biscuits and pastries, bread, ice-cream chocolate and confectionery, savory pastries and sandwiches), 37.2% only industrial (ice-cream, chocolate and confectionery, cakes, biscuits and pastries, snacks, bread and savory pastries) and 10.1% both. Homemade foods presented significantly higher total fat [homemade 11.6 g (range 6.6-19.4 g); industrial 6.2 g (range 4.0-8.6 g), p = 0.001], monounsaturated, polyunsaturated and trans-fat, and sodium and potassium content per serving. Industrial wafers presented the highest mean saturated (11.8 g/serving) and trans-fat (2.32 g/serving) content. Homemade hamburgers presented the highest mean sodium content (1889 mg/serving). Conclusions: Strategies to encourage the production and sales of healthier street foods, especially homemade, are needed to promote healthier urban food environments in urban Turkmenistan.
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Bebidas , Bocadillos , Valor Nutritivo , Sodio , TurkmenistánRESUMEN
Street food makes a significant contribution to the diet of many dwellers in low- and middle-income countries and its trade is a well-developed activity in the central Asian region. However, data on its purchase and nutritional value is still scarce. This study aimed to describe street food purchasing patterns in central Asia, according to time and place of purchase. A multicentre cross-sectional study was conducted in 2016/2017 in the main urban areas of four central Asian countries: Dushanbe (Tajikistan), Bishkek (Kyrgyzstan), Ashgabat (Turkmenistan) and Almaty (Kazakhstan). Street food markets (n = 34) and vending sites (n = 390) were selected by random and systematic sampling procedures. Data on the purchased foods and beverages were collected by direct observation. Time and geographic location of the purchases was registered, and their nutritional composition was estimated. A total of 714 customers, who bought 852 foods, were observed. Customers' influx, buying rate and purchase of industrial food were higher in city centers compared to the outskirts (median: 4.0 vs. 2.0 customers/10 min, p < 0.001; 5.0 vs. 2.0 food items/10 min, p < 0.001; 36.2 vs. 28.7%, p = 0.004). Tea, coffee, bread and savory pastries were most frequently purchased in the early morning, bread, main dishes and savory pastries during lunchtime, and industrial products in the mid-morning and mid-afternoon periods. Energy and macronutrient density was highest at 11:00-12:00 and lowest at 09:00-10:00. Purchases were smaller but more energy-dense in city centers, and higher in saturated and trans-fat in the peripheries. This work provides an overview of the street food buying habits in these cities, which in turn reflect local food culture. These findings from the main urban areas of four low- and middle-income countries which are currently under nutrition transition can be useful when designing public health interventions customized to the specificities of these food environments and their customers.
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Glioma is characterized by an active production of proangiogenic molecules. We observed that conditioned medium (CM) from C6 glioma significantly enhanced proliferation and migration of immortalized rat brain GP8.3 endothelial cells (ECs) and primary bovine brain microvascular ECs. The glioma CM effect was significantly reduced by cytosolic (cPLA(2)) and Ca(++)-independent (iPLA(2)) phospholipase A(2), cyclooxygenase-2, and protein kinase inhibitors. In GP8.3 ECs, cPLA(2) and iPLA(2) enzyme activities and phosphorylation of cPLA(2), significantly stimulated after 24h CM co-incubation, were attenuated by PLA(2), PI3-K, MEK-1, and ERK1/2 inhibitors. By confocal microscopy, in glioma CM-stimulated ECs, enhancement of fluorescence signals for phospho-cPLA(2), phospho-ERK1/2, phospho-PKCα, COX-2, and iPLA(2) was in parallel observed. Electroporation of anti-iPLA(2) and cPLA(2) antibodies and siRNAs directed against iPLA(2) and cPLA(2) significantly inhibited cell proliferation and migration. Incubation of CM- or VEGF peptide-stimulated ECs with antibodies against VEGF or VEGFR-1/-2 receptors strongly reduced mitotic rate, cell migration, and phospho-cPLA(2) and iPLA(2) protein levels. The findings suggest that PLA(2) activities are involved in stimulating EC migration and proliferation in the presence of glioma CM and that cPLA(2) is positively regulated upstream by PI3-K, PKCα, and ERK1/2 signal cascades. Our work provides new insights in understanding EC metabolism and signaling during tumor angiogenesis.
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Encéfalo/citología , Células Endoteliales/enzimología , Glioma/metabolismo , Neovascularización Patológica/metabolismo , Fosfolipasas A2 Calcio-Independiente/metabolismo , Fosfolipasas A2 Citosólicas/metabolismo , Animales , Anticuerpos/administración & dosificación , Anticuerpos/inmunología , Anticuerpos/farmacología , Bovinos , Línea Celular Transformada , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Medios de Cultivo Condicionados/farmacología , Ciclooxigenasa 1/metabolismo , Ciclooxigenasa 2/metabolismo , Electroporación , Células Endoteliales/citología , Células Endoteliales/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Expresión Génica/efectos de los fármacos , Expresión Génica/genética , Glioma/irrigación sanguínea , Espacio Intracelular/metabolismo , Proteínas de la Membrana/metabolismo , Proteína Quinasa 1 Activada por Mitógenos/antagonistas & inhibidores , Proteína Quinasa 1 Activada por Mitógenos/metabolismo , Proteína Quinasa 3 Activada por Mitógenos/antagonistas & inhibidores , Proteína Quinasa 3 Activada por Mitógenos/metabolismo , Modelos Biológicos , Fosfolipasas A2 Calcio-Independiente/antagonistas & inhibidores , Fosfolipasas A2 Calcio-Independiente/genética , Fosfolipasas A2 Calcio-Independiente/inmunología , Fosfolipasas A2 Citosólicas/antagonistas & inhibidores , Fosfolipasas A2 Citosólicas/genética , Fosfolipasas A2 Citosólicas/inmunología , Fosforilación/efectos de los fármacos , Proteína Quinasa C-alfa/antagonistas & inhibidores , Proteína Quinasa C-alfa/metabolismo , Inhibidores de Proteínas Quinasas/farmacología , ARN Interferente Pequeño/genética , Ratas , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Factor A de Crecimiento Endotelial Vascular/antagonistas & inhibidores , Factor A de Crecimiento Endotelial Vascular/inmunología , Factor A de Crecimiento Endotelial Vascular/farmacologíaRESUMEN
This study aimed to describe street food purchases in cities from Central Asia, considering customers' characteristics and the nutritional composition of the foods and beverages. Cross-sectional studies were conducted in 2016/2017 in Dushanbe (Tajikistan), Bishkek (Kyrgyzstan), Ashgabat (Turkmenistan) and Almaty (Kazakhstan). Direct observation was used to collect data on the purchases made by street food customers, selected by random and systematic sampling. Nutritional composition was estimated using data from chemical analyses, food composition tables or food labels. A total of 714 customers (56.6% females, 55.5% aged ≥35 years, 23.3% overweight/obese) were observed, who bought 852 foods and beverages, the most frequent being savoury pastries/snacks (23.2%), main dishes (19.0%), sweet pastries/confectionery (17.9%), tea/coffee (11.3%) and soft drinks/juices (9.8%). Fruit was the least purchased food (1.1%). Nearly one-third of customers purchased industrial food items (31.9%). The median energy content of a street food purchase was 529 kcal/serving. Saturated and trans-fat median contents were 4.7 g/serving and 0.36 g/serving (21.4% and 16.5% of maximum daily intake recommendations, respectively). Median sodium and potassium contents were 745 mg/serving (37.3% of maximum recommendation) and 304 mg/serving (8.7% of minimum recommendation), respectively. In general, the purchases observed presented high contents of energy, saturated-fat, trans-fat and sodium, and low levels of potassium. Policies towards the improvement of these urban food environments should be encouraged.
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Alimentos , Población Urbana , Adulto , Asia , Bebidas , Peso Corporal , Estudios Transversales , Femenino , Humanos , Masculino , Valor NutritivoRESUMEN
The seasonal variation in chemical and lipid composition of chub mackerel (Scomber colias) was evaluated monthly over one year (proximate chemical composition and lipid profile: fatty acid (FA) and lipid classes distribution). Significant seasonal changes regarding fat content were noticed (1.3-10.3 g/100 g), with the lowest fat content obtained in February (during spawning period), and the highest in September. Regarding the FA profile, the main fluctuations were recorded in saturated (SFA) and polyunsaturated fatty acids (PUFA). The highest SFA content was registered between March and August (25.3-32.3%). PUFA (the most abundant group) reached its maximum percentual content between December and February (60.9 and 66.9%, respectively). In absolute terms, PUFA attained 5352.6 mg/100 g edible part in September, where 2473.8 mg/100 g of docosahexaenoic acid (DHA, C22:6n-3), representing 46.2% of total PUFA. DHA lowest level was 519.8 mg/100 g, registered in low-fat chub mackerel. Together DHA and EPA (eicosapentaenoic acid, C20:5n-3) represented 75% of the total PUFA and 84% of n-3 PUFA. Triacylglycerols (TAG) with 82.2-92.1% of total lipid content) and phospholipids (4.4-8.4%) were the main lipid classes. Polar lipid fraction (phospholipids), was predominantly constituted by PUFA (68.6-74.5%), mainly DHA (45.2-55.1%), with the highest percentage recorded in low-fat chub mackerel. High relative contents of PUFA (36.6-49.1%) were also found in TAG. Having into account the data obtained, chub mackerel is a privileged source of DHA even in a lean species whereby its consumption should be recommended as part a healthy dietary regime.
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Lípidos/análisis , Lípidos/química , Valor Nutritivo , Perciformes , Estaciones del Año , Animales , Ácidos Grasos/análisis , Proteínas de Peces/análisis , Análisis de los Alimentos , Humanos , Perciformes/metabolismo , Fosfolípidos/análisis , Triglicéridos/análisisRESUMEN
Quantification of the specific folate vitamers to estimate total folate in foods is not standardized. A collaborative study, including eight European laboratories, was conducted in order to determine the repeatability and reproducibility of the method for folate quantification in foods using the plant-origin γ-glutamyl hydrolase as part of the extraction procedure. The seven food samples analyzed represent the food groups; fruits, vegetables, dairy products, legumes, offal, fish, and fortified infant formula. The homogenization step was included, and six folate vitamers were analyzed using LC-MS/MS. Total folate content, expressed as folic acid equivalent, was 17-490 µg/100 g in all samples. Horwitz ratio values were within the acceptable range (0.60-1.94), except for fish. The results for fortified infant formula, a certified reference material (NIST 1869), confirmed the trueness of the method. The collaborative study is part of a standardization project within the Nordic Committee on Food Analysis (NMKL).
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Fraccionamiento Químico/métodos , Ácido Fólico/análisis , Análisis de los Alimentos/métodos , Espectrometría de Masas en Tándem/métodos , Animales , Cromatografía Liquida/métodos , Cromatografía Liquida/normas , Productos Lácteos/análisis , Grano Comestible/química , Productos Pesqueros/análisis , Análisis de los Alimentos/normas , Alimentos Fortificados/análisis , Frutas/química , Humanos , Lactante , Fórmulas Infantiles/análisis , Reproducibilidad de los Resultados , Espectrometría de Masas en Tándem/normas , Verduras/químicaRESUMEN
We provide novel evidence that human melanoma cell lines (M10, M14, SK-MEL28, SK-MEL93, 243MEL, 1074MEL, OCM-1, and COLO38) expressed, at mRNA and protein levels, either Ca(2+)-independent phospholipase A(2) (iPLA(2)) or cytosolic phospholipase A(2) (cPLA(2)) and its phosphorylated form. Normal human melanocytes contained the lowest levels of both PLA(2)s. Cyclooxygenase-1 and -2 (COX-1 and COX-2) were also expressed in cultured tumor cells as measured by Western blots. The most pronounced overexpression of iPLA(2) and COX-1 was found in two melanoma-derived cells, M14 and COLO38. Normal human melanocytes and the M10 melanoma cell line displayed no COX-2 expression. Using subcellular fractionation, Western blot and confocal microcopy analyses, in paradigmatic SK-MEL28 and SK-MEL93 cells we showed that iPLA(2), COX-1 and even cPLA(2) were equally located in the cytosol, membrane structures and perinuclear region while COX-2 was preferentially associated with the cytosol. Specific inhibitors of these three enzymes significantly reduced the basal proliferation rate either in melanocytes or in melanoma cell lines. These results, coupled with the inhibition of the cell proliferation by electroporation of melanoma cells with cPLA(2) or COX-2 antibodies, demonstrate that a possible correlation between PLA(2)-COX expression and tumor cell proliferation in the melanocytic system does exist. In addition, the high expression level of both PLA(2)s and COXs suggests that eicosanoids modulate cell proliferation and tumor invasiveness.
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Melanocitos/metabolismo , Fosfolipasas A2 Calcio-Independiente/metabolismo , Fosfolipasas A2/metabolismo , Prostaglandina-Endoperóxido Sintasas/metabolismo , Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/farmacología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Ciclooxigenasa 1/genética , Ciclooxigenasa 1/metabolismo , Ciclooxigenasa 2/inmunología , Ciclooxigenasa 2/metabolismo , Inhibidores de la Ciclooxigenasa 2/farmacología , Citosol/metabolismo , Inhibidores Enzimáticos/farmacología , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Fosfolipasas A2 Grupo II/genética , Fosfolipasas A2 Grupo II/metabolismo , Humanos , MAP Quinasa Quinasa 1/antagonistas & inhibidores , Melanocitos/citología , Melanocitos/enzimología , Melanoma/enzimología , Melanoma/genética , Melanoma/patología , Orgánulos/metabolismo , Inhibidores de las Quinasa Fosfoinosítidos-3 , Inhibidores de Fosfolipasa A2 , Fosfolipasas A2/genética , Fosfolipasas A2 Calcio-Independiente/antagonistas & inhibidores , Fosfolipasas A2 Calcio-Independiente/genética , Fosforilación/efectos de los fármacos , Prostaglandina-Endoperóxido Sintasas/genética , Prostaglandina-Endoperóxido Sintasas/inmunología , Inhibidores de Proteínas Quinasas/farmacologíaRESUMEN
The largely undefined signal transduction mechanisms and cross-talk between human melanoma cell (HMC) lines and brain endothelial cells (ECs) involved in tumor cell interaction and adhesion were investigated. In immortalized rat brain GP8.3 EC cultures, conditioned media (CM) prepared from SK-MEL28 and OCM-1 melanoma cells significantly enhanced arachidonic acid release, cytosolic phospholipase A(2) (cPLA(2)) and Ca(+)-independent phospholipase A(2) (iPLA(2)) specific activities, and cell growth by 24 h. Inhibitors such as wortmannin and LY294002 (vs. PI3 kinase activity), AACOCF(3), (vs. cPLA(2) and iPLA(2)), PD98059 (vs. ERK1/2 activity) and NS-398 (vs. cyclooxygenase-2 activity, COX-2) were all able to block cell proliferation and motility determined using a scratch wound healing assay in melanoma CMs-stimulated EC monolayers. These media also support the enhanced cell proliferation of primary ECs derived from rat brain (BBEC). Electroporation of anti-cPLA(2) antibody into ECs markedly inhibited the EC proliferation in response to CMs. With both CMs, phosphorylation of cPLA(2), PKCalpha, ERK1/2, protein and mRNA expression of cPLA(2) and iPLA(2), and COX-2 protein expression were significantly stimulated after 24 h coincubation, and attenuated by specific inhibitors. By confocal microscopy, activation of cPLA(2), ERK1/2, PKCalpha and COX-2 in perinuclear and membrane regions of ECs grown in CM-stimulated cultures were clearly observed. Thus MEK-PKCalpha-ERK1/2 and PI3-K/Akt survival pathways are activated in EC cultures during the interaction with CM from both melanoma cell lines, providing new insight in understanding EC metabolism and signaling. These pathways represent potential therapeutic targets to inhibit or enhance tumor angiogenesis.
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Células Endoteliales/citología , Melanoma/metabolismo , Neovascularización Patológica/enzimología , Fosfolipasas A2/metabolismo , Proteína Quinasa C-alfa/metabolismo , Anticuerpos Bloqueadores/farmacología , Encéfalo/irrigación sanguínea , Línea Celular Transformada , Línea Celular Tumoral , Membrana Celular/efectos de los fármacos , Membrana Celular/enzimología , Movimiento Celular , Proliferación Celular/efectos de los fármacos , Medios de Cultivo Condicionados/farmacología , Electroporación , Células Endoteliales/fisiología , Endotelio Vascular/citología , Humanos , Sistema de Señalización de MAP Quinasas , Proteína Quinasa C-alfa/genética , Proteína Quinasa C-alfa/inmunología , Transducción de SeñalRESUMEN
Despite being highly appreciated and consumed, the nutritional value of Chaceon maritae from Namibe (Angola) had never been studied. In the present work, edible tissues (muscle, ovaries, and hepatopancreas) of boiled female C. maritae caught off Namibe coast in two distinct seasons were analyzed in terms of proximate chemical composition (fat, ash, protein, and moisture), fatty acid and amino acid profiles, cholesterol, essential minerals (macro and trace) and toxic elements. Results showed that, in both seasons, C. maritae muscle was a valuable source of protein, essential amino acids, polyunsaturated fatty acids, and essential elements, especially zinc, selenium, iodine, and copper. Ovaries and hepatopancreas are also good sources of protein, but were richer in fat, particularly when caught in October. Ovarian fat is rich in polyunsaturated fatty acids and that of hepatopancreas has higher values of monounsaturated and saturated fatty acids. Hepatopancreas and ovaries are also good sources of copper and, especially ovaries, of zinc. Moreover, in both seasons, all the edible tissues of C. maritae analyzed presented very low contents of heavy metals (mercury, cadmium, lead, and arsenic).
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Industrial cooking of common octopus (Octopus vulgaris) under well-established procedures is advantageous for current consumers, which demand healthy and convenient food. This work aimed to evaluate the effect of industrial water boiling, without the addition of salt, on the nutritional profile of common octopus. True retentions (TRs) were calculated for essential nutrients and toxic elements. After boiling, the moisture content decreased, resulting in a concentration of other constituents (protein, fat, fatty acids, majority of amino acids, phosphorus, zinc, and iodine). High TRs were obtained for some essential nutrients: 90.2% (eicosapentaenoic acid, EPA), 89.1% (docosahexaenoic acid, DHA), ≥74.6% (indispensable amino acids, IAA), and 86.8% (iodine). In both raw and boiled octopus, polyunsaturated fatty acids (252.2 and 425.1 mg/100 g), leucine (940.1 and 1613.4 mg/100 g), glutamate (1971.5 and 3257.1 mg/100 g), sodium (393.3 and 332.5 mg/100 g), and zinc (12.6 and 16.6 mg/kg) were, respectively, the most abundant fatty acids, IAA, dispensable amino acids, macro, and micro elements. Cadmium, lead, and mercury levels found in boiled octopus were 0.02, 0.10, and 0.08 mg/kg, respectively. The consumption of 150 g (usual portion) of boiled octopus is advisable because it contributes to significant daily intakes of EPA+DHA (>100%), selenium (75.6%), and iodine (12.4%), and 25% of the daily adequate intake of sodium for adults.
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This study focused on the use of a new system, an alginate|Æ-poly-l-lysine|alginate|chitosan microcapsule (APACM), able to immobilize a folate-producing probiotic, Lactococcus lactis ssp. cremoris (LLC), which provides a new approach to the utilization of capsules and probiotics for in situ production of vitamins. LLC is able to produce 95.25±26µg·L-1 of folate, during 10h, and was encapsulated in the APACM. APACM proved its capacity to protect LLC against the harsh conditions of a simulated digestion maintaining a viable concentration of 6logCFU·mL-1of LLC. A nutrients exchange capacity test, was performed using Lactobacillus plantarum UM7, a high lactic acid producer was used here to avoid false negative results. The production and release of 2g·L-1 of lactic acid was achieved through encapsulation of L. plantarum, after 20h. The adhesion of APACM to epithelial cells was also quantified, yielding 38% and 33% of capsules adhered to HT-29 cells and Caco-2 cells, respectively.
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PURPOSE: To investigate the molecular mechanisms of the antiangiogenic activity of UPARANT, an antagonist of the urokinase-type plasminogen activator receptor (uPAR), on primary human retinal endothelial cells (HREC) as a model of in vitro angiogenesis. METHODS: The antiangiogenic activity of UPARANT was evaluated on endothelial cell migration, invasion, and tube formation. Human REC were further analyzed for viability, transendothelial electrical resistance (TEER), and tight junction (TJ) expression at the protein and mRNA levels. Vascular endothelial growth factor-related signaling molecules were also analyzed by Western and northern blots. RESULTS: UPARANT inhibited in a dose-dependent fashion HREC motility, invasion, and tube formation stimulated by VEGF-A, in a range of doses (1-100 nM) that had no effect on cell viability and proliferation. UPARANT also prevented the loss of permeability induced by VEGF-A, restoring normal TEER values and TJ protein expression. At the molecular level, UPARANT inhibited VEGFR-2 and STAT3 phosphorylation, thus decreasing VEGF and hypoxia-inducible factor 1-alpha expression, finally resulting in decreased activation of MEK/ERK, JNK, p38, and AKT signaling proteins. CONCLUSIONS: These findings indicate that UPARANT exerts its antiangiogenic effects through the inhibition of the downstream signaling activated by angiogenic factors such as VEGF-A.
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Células Endoteliales/metabolismo , Lectinas de Unión a Manosa/metabolismo , Glicoproteínas de Membrana/metabolismo , Receptores de Superficie Celular/metabolismo , Retina/patología , Neovascularización Retiniana/metabolismo , Western Blotting , Recuento de Células , Movimiento Celular , Proliferación Celular , Células Cultivadas , Células Endoteliales/patología , Humanos , Retina/metabolismo , Neovascularización Retiniana/patología , Transducción de Señal , Receptor 2 de Factores de Crecimiento Endotelial Vascular/metabolismoRESUMEN
Primary solid tumors originate close to pre-existing tissue vasculature, initially growing along such tissue blood vessels, and this phenomenon is important for the metastatic potential which frequently occurs in highly vascularized tissues. Unfortunately, preclinic and clinic anti-angiogenic approaches have not been very successful, and multiple factors have been found to contribute to toxicity and tumor resistance. Moreover, tumors can highlight intrinsic or acquired resistances, or show adaptation to the VEGF-targeted therapies. Furthermore, different mechanisms of vascularization, activation of alternative signaling pathways, and increased tumor aggressiveness make this context even more complex. On the other hand, it has to be considered that the transitional restoration of normal, not fenestrated, microvessels allows the drug to reach the tumor and act with the maximum efficiency. However, these effects are time-limited and different, depending on the various types of cancer, and clearly define a specific "normalization window." So, new horizons in the therapeutic approaches consist on the treatment of the tumor with pro- (instead of anti-) angiogenic therapies, which could strengthen a network of well-structured blood vessels that facilitate the transport of the drug.
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It is suspected that microbial infections take part in the pathogenesis of diabetes mellitus type 1 (T1DM). Glucose-induced insulin secretion is accompanied by the release of free arachidonic acid (AA) mainly by cytosolic- and calcium independent phospholipases A2 (cPLA2 and iPLA2). Insulinoma cell line (INS-1E) was infected with E. coli isolated from the blood culture of a patient with sepsis. Invasion assay, Scanning Electron Microscopy and Transmission Electron Microscopy demonstrated the capacity of E. coli to enter cells, which was reduced by PLA2 inhibitors. Glucose-induced insulin secretion was significantly increased after acute infection (8h) but significantly decreased after chronic infection (72h). PLA2 activities, cPLA2, iPLA2, phospho-cPLA2, and COX-2 expressions were increased after acute and, even more, after chronic E. coli infection. The silencing of the two isoforms of PLA2s, with specific cPLA2- or iPLA2-siRNAs, reduced insulin secretion after acute infection and determined a rise in insulin release after chronic infection. Prostaglandins E2 (PGE2) production was significantly elevated in INS-1E after long-term E. coli infection and the restored insulin secretion in presence of L798106, a specific EP3 antagonist, and NS-398, a COX-2 inhibitor, and the reduction of insulin secretion in presence of sulprostone, a specific EP3 agonist, revealed their involvement in the effects triggered by bacterial infection. The results obtained demonstrated that cPLA2 and iPLA2 play a key role in insulin secretion process after E. coli infection. The high concentration of AA released is transformed into PGE2, which could be responsible for the reduced insulin secretion.
Asunto(s)
Calcio/metabolismo , Citosol/metabolismo , Dinoprostona/metabolismo , Escherichia coli/metabolismo , Insulina/metabolismo , Fosfolipasas A2/metabolismo , Línea Celular , Activación Enzimática , Humanos , Secreción de InsulinaRESUMEN
Inhibitors of PARP-1(Poly(ADP-ribose) polymerase-1) act by competing with NAD(+), the enzyme physiological substrate, which play a protective role in many pathological conditions characterized by PARP-1 overactivation. It has been shown that PARP-1 also promotes tumor growth and progression through its DNA repair activity. Since angiogenesis is an essential requirement for these activities, we sought to determine whether PARP inhibition might affect rat brain microvascular endothelial cells (GP8.3) migration, stimulated by C6-glioma conditioned medium (CM). Through wound-healing experiments and MTT analysis, we demonstrated that PARP-1 inhibitor PJ-34 [N-(6-Oxo-5,6-dihydrophenanthridin-2-yl)-N,N-dimethylacetamide] abolishes the migratory response of GP8.3 cells and reduces their viability. PARP-1 also acts in a DNA independent way within the Extracellular-Regulated-Kinase (ERK) signaling cascade, which regulates cell proliferation and differentiation. By western analysis and confocal laser scanning microscopy (LSM), we analyzed the effects of PJ-34 on PARP-1 expression, phospho-ERK and phospho-Elk-1 activation. The effect of MEK (mitogen-activated-protein-kinase-kinase) inhibitor PD98059 (2-(2-Amino-3-methoxyphenyl)-4 H-1-benzopyran-4-one) on PARP-1 expression in unstimulated and in CM-stimulated GP8.3 cells was analyzed by RT-PCR. PARP-1 expression and phospho-ERK activation were significantly reduced by treatment of GP8.3 cells with PJ-34 or PD98059. By LSM, we further demonstrated that PARP-1 and phospho-ERK are coexpressed and share the same subcellular localization in GP8.3 cells, in the cytoplasm as well as in nucleoplasm. Based on these data, we propose that PARP-1 and phospho-ERK interact in the cytosol and then translocate to the nucleus, where they trigger a proliferative response. We also propose that PARP-1 inhibition blocks CM-induced endothelial migration by interfering with ERK signal-transduction pathway.
Asunto(s)
Neoplasias Encefálicas/metabolismo , Encéfalo/irrigación sanguínea , Medios de Cultivo Condicionados/metabolismo , Células Endoteliales/efectos de los fármacos , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Glioma/metabolismo , Microvasos/efectos de los fármacos , Comunicación Paracrina/efectos de los fármacos , Fenantrenos/farmacología , Inhibidores de Poli(ADP-Ribosa) Polimerasas/farmacología , Poli(ADP-Ribosa) Polimerasas/metabolismo , Transporte Activo de Núcleo Celular/efectos de los fármacos , Animales , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Células Endoteliales/enzimología , Quinasas MAP Reguladas por Señal Extracelular/antagonistas & inhibidores , Microvasos/enzimología , Fosforilación , Poli(ADP-Ribosa) Polimerasa-1 , Poli(ADP-Ribosa) Polimerasas/genética , Unión Proteica , Inhibidores de Proteínas Quinasas/farmacología , Ratas , Transducción de Señal/efectos de los fármacos , Proteína Elk-1 con Dominio ets/metabolismoRESUMEN
Retinoblastoma is the most common intraocular tumour in children. In view of understanding the molecular mechanisms through which angiogenic switch on happens in the early phases of reciprocal interaction between tumour and cells constituting retinal microvessel, Transwell co-cultures constituted by human retinal endothelial cells (HREC), pericytes (HRPC), and human retinoblastoma cell line Y-79 were performed. Y-79 enhanced HREC proliferation, reduced by the introduction of HRPC in triple culture. In HREC/HRPC cultures, TGF-ß in media increased, decreasing in triple cultures. High VEGF levels in triple cultures witnessed the establishment of a strongly in vitro angiogenic environment. Y-79 induced in HREC an increase in c- and iPLA2, phospho-cPLA2, inducible COX-2 protein expressions, PLA2 activities and prostaglandin E2 (PGE2) release. These effects were attenuated when HRPC were introduced in triple culture. Moreover, antibody silencing of TGF-ß demonstrated a strong correlation between the signalling pathway triggered by TGF-ß of pericytal origin and the phospholipase activation and the modulation of PGE2 release. Inhibiting VEGFA effect, the HRPC loss in triple culture decreased, showing its modulatory effect on their survival. Relying on the data here presented, sustaining the pericytal survival in a tumour retinal environment could ensure the integrity of microvessels and the TGF-ß supply, essential for controlling aberrant endothelial pruning and angiogenesis.