Beneficios de los espacios verdes y actividad física en el bienestar y salud de las personas / Benefits of green spaces and physical activity for the well-being and health of people

BACKGROUND: Physical environment can influence well-being, quality of life and health and population health in several ways. Exposure to green spaces improves the physical and mental health of individuals. Chile has privileged conditions for outdoor activities that could benefit millions of people. However, a small percentage of the Chilean population is exposed to green spaces in the recommended amounts to promote health. AIM: To describe the benefits of green spaces on physical and mental health and their relationship with the practice of physical activity. MATERIAL AND METHODS: Review of the scientific literature in English in the Web of Science (WoS) electronic database published between 2006-2019. RESULTS: In addition to the direct benefits of green spaces, there are synergistic effects of performing physical activities in these environments such as feelings of good health, satisfaction with life and pleasure, increased physiological relaxation, positive emotions, mental well-being, restoration of attention, decreased perceived stress and a reduction negative affections. CONCLUSIONS: This review supports strategies for improving access to green spaces in urban areas coupled with physical activity promotion in these settings. Health and urban planning stakeholders should consider these aspects in future programs.

[Benefits of green spaces and physical activity for the well-being and health of people]. / Beneficios de los espacios verdes y actividad física en el bienestar y salud de las personas.

BACKGROUND: Physical environment can influence well-being, quality of life and health and population health in several ways. Exposure to green spaces improves the physical and mental health of individuals. Chile has privileged conditions for outdoor activities that could benefit millions of people. However, a small percentage of the Chilean population is exposed to green spaces in the recommended amounts to promote health. AIM: To describe the benefits of green spaces on physical and mental health and their relationship with the practice of physical activity. MATERIAL AND METHODS: Review of the scientific literature in English in the Web of Science (WoS) electronic database published between 2006-2019. RESULTS: In addition to the direct benefits of green spaces, there are synergistic effects of performing physical activities in these environments such as feelings of good health, satisfaction with life and pleasure, increased physiological relaxation, positive emotions, mental well-being, restoration of attention, decreased perceived stress and a reduction negative affections. CONCLUSIONS: This review supports strategies for improving access to green spaces in urban areas coupled with physical activity promotion in these settings. Health and urban planning stakeholders should consider these aspects in future programs.

Associations Between Movement Behaviors and Emotional Changes in Toddlers and Preschoolers During Early Stages of the COVID-19 Pandemic in Chile.

Background: There is limited evidence about emotional and behavioral responses in toddlers and preschoolers during the novel coronavirus (COVID-19) pandemic, particularly in Latin America. Objective: To assess associations between changes in movement behaviors (physical activity, screen time and sleeping) and emotional changes in toddlers and preschoolers during early stages of the pandemic in Chile. Methods: A cross-sectional study conducted from March 30th to April 27th, 2020. Main caregivers of 1- to 5-year-old children living in Chile answered an online survey that included questions about sociodemographic characteristics, changes in the child's emotions and behaviors, movement behaviors and caregivers' stress during the pandemic. Multiple linear regressions were used to assess the association between different factors and emotional changes in toddlers and preschoolers. Results: In total, 1727 caregivers provided complete data on emotional changes for children aged 2.9 ± 1.36 years old, 47.9% girls. A large proportion of toddlers and preschoolers in Chile experienced emotional and behavioral changes. Most caregivers reported that children "were more affectionate" (78.9%), "more restless" (65.1%), and 'more frustrated' (54.1%) compared with pre-pandemic times. Apart from changes in movement behaviors, factors such as child age, caregivers' age and stress, and residential area (urban/rural) were consistently associated with changes in emotions and behaviors. Conclusion: The pandemic substantially affected the emotions and behaviors of toddlers and preschoolers in Chile. The findings suggest that supportive actions for caregivers may have a positive impact not only on adults but also on children. Mental health promotion programs should consider multilevel approaches in which the promotion of movement behaviors and support for caregivers should be essential pieces for future responses.

Cuidados enfermeros que modifican conductas alimentarias en adolescentes, etapa inicial de anorexia / Nursing cares that modify eating behaviors in adolescents, first stage of anorexia / Cuidados de enfermagem que mudam comportamentos alimentares em adolescentes, etapa inicial da anorexia

Introducción: La anorexia nerviosa es un trastorno de la conducta alimentaria que afecta al individuo fisiológica y psicológicamente. El profesional de enfermería y equipo multidisciplinar debe promocionar, prevenir y rehabilitar para modificar estas conductas y reducir su progresión. Metodología: Revisión integrativa de la literatura. Palabras clave utilizadas Atención de Enfermería, Jóvenes, Anorexia, Conducta alimentaria trastorno de la Ingesta Alimentaria Evitativa/Restrictiva, Tratamiento multimodal y Habilidades de afrontamiento. Se incluyeron artículos en español, inglés y portugués de cualquier nacionalidad y diseño metodológico publicados entre 2014 y 2019 indexados en bases de datos ClinicalKey, Scielo, CINAHL y metabuscador Google académico. Se filtraron por lectura de título, resumen y texto completo, se clasificaron según el nivel de evidencia y grado de recomendación; finalmente se ejecutó lectura crítica mediante las escalas Amstar, Consort y Strobe.. Resultados: Se incluyeron 34 artículos y se construyeron cuatro temáticas: cuidados de enfermería que modifican la conducta alimentaria, factores que influyen en el cambio de la conducta alimentaria, impacto de los cuidados de enfermería y consecuencias de la no aplicación de los cuidados de enfermería. Conclusión: Las intervenciones de enfermería son fundamentales para la recuperación de los adolescentes en etapa inicial de anorexia ya que reducen la progresión de la enfermedad y favorece la expresión de sentimientos.

Introduction: Anorexia nervosa is an eating disorder that affects the individual physiologically and psychologically. The nursing professional, along with a multidisciplinary team must promote, prevent and rehabilitate the individual, to modify these behaviors and reduce its progression. Methodology: Integrative review of the literature. Keywords used: nursing care, young people, anorexia, eating behavior, avoidant/restrictive food intake disorders, multimodal treatment and methodological design. Articles in English, Spanish and Portuguese were included, from any nationality and methodological design. The articles selected were published between 2014 and 2019 and were indexed to the databases ClinicalKey, Scielo, CINAHL, and Google Scholar. These were filtered by reading the title, abstract and complete text; and were classified according to their level of evidence and grades of recommendation. Finally, a critical reading was made using the tools, AMSTAR, CONSORT and STROBE. Results: 34 articles were included, and four themes were built: nursing care to modify eating habits, factors that include change in eating behaviors, nursing care impact and consequences of not applying nursing care. Conclusion: The nursing interventions are fundamental for the recovery of adolescents in the first stage of anorexia since these reduce the progression of the disease and support expressing their feelings.

Introdução: A anorexia nervosa é um transtorno do comportamento alimentar que afeta ao sujeito fisiologica e psicologicamente. O profissional de enfermagem junto com a equipe multiprofissional deve promover, prever e reabilitar para mudar essas condutas e reduzir o progresso. Métodos: Revisão integrativa da literatura. Palavras-chave empregadas: cuidados de enfermagem, adolescente, anorexia, comportamento alimentar, transtorno da evitação ou restrição da ingestão de alimentos, terapia combinada, adaptação psicológica. Incluíram-se artigos publicados em espanhol, inglês e português com qualquer nacionalidade ou metodologia. Foram selecionadas as publicações entre 2014-2019 disponíveis na ClinicalKey, Scielo, CINAHL e no Google Scholar. Foram filtrados pela leitura do título, o resumo e o texto completo; classificaram-se segundo o nível de evidência e o grau de recomendação. Finalmente, efetivou-se uma leitura crítica empregando as escalas Amstar, Consort e Strobe. Resultados: Incluíram-se 34 artigos e construíram-se quatro temas: cuidados de enfermagem que mudam os comportamentos alimentares; fatores que influenciam a mudança de comportamento alimentar; impacto nos cuidados de enfermagem; e, consequências da ausência de cuidados de enfermagem. Conclusão: As intervenções de enfermagem são fundamentais para a recuperação dos adolescentes na etapa inicial da anorexia, devido à redução do progresso do transtorno, além de favorecer a expressão dos seus sentimentos.

Palmitate decreases migration and proliferation and increases oxidative stress and inflammation in smooth muscle cells: role of the Nrf2 signaling pathway.

Fatty acids are essential to cell functionality and may exert diverging vascular effects including migration, proliferation, oxidative stress, and inflammation. This study examined the effect of palmitate on human coronary artery smooth muscle cell (HCASMC) function. An in vitro wound-healing assay indicated that palmitate decreased HCASMC migration in dose- and time-dependent manners. Furthermore, bromodeoxyuridine incorporation assays indicated that palmitate decreased HCASMC proliferation in a dose-response manner. Palmitate also increased reactive oxygen species formation, malondialdehyde content, and intracellular lipid droplets accompanied with increased fatty acid binding protein 4 expression. Moreover, palmitate induced gene expression (monocyte chemoattractant protein 1, matrix metalloproteinase-2, IL-1ß, IL-6, IL-8, and TNF-α) and intracellular protein content (plasminogen activator inhibitor-1 and urokinase plasminogen activator) of inflammatory mediators. Finally, we showed that palmitate activates the transcription factor Nrf2 and the upstream kinases ERK1/2 and Akt in HCASMCs. The inhibitor of Nrf2, trigonelline, significantly attenuated palmitate-induced HCASMC expression of the Nrf2 target gene NQO1. These findings indicate that palmitate might be critically related to HCASMC function by slowing cell migration and proliferation and inducing lipid-laden cells, oxidative stress, and inflammation in part by activation of the Nrf2 transcription factor. Palmitate's activation of proinflammatory Nrf2 signaling may represent a novel mechanism mediating the proatherogenic actions of saturated fatty acids.

Food additives, contaminants and other minor components: effects on human gut microbiota-a review

Gut bacteria play an important role in several metabolic processes and human diseases, such as obesity and accompanying co-morbidities, such as fatty liver disease, insulin resistance/diabetes, and cardiovascular events. Among other factors, dietary patterns, probiotics, prebiotics, synbiotics, antibiotics, and non-dietary factors, such as stress, age, exercise, and climatic conditions, can dramatically impact the human gut microbiota equilibrium and diversity. However, the effect of minor food constituents, including food additives and trace contaminants, on human gut microbiota has received less attention. Consequently, the present review aimed to provide an objective perspective of the current knowledge regarding the impacts of minor food constituents on human gut microbiota and consequently, on human health

Influence of food consumption patterns and Galician lifestyle on human gut microbiota

The proportion of different microbial populations in the human gut is an important factor that in recent years has been linked to obesity and numerous metabolic diseases. Because there are many factors that can affect the composition of human gut microbiota, it is of interest to have information about what is the composition of the gut microbiota in different populations in order to better understand the possibilities for improving nutritional management. A group of 31 volunteers were selected according to established inclusion and exclusion criteria and were asked about their diet history, lifestyle patterns, and adherence to the Southern European Atlantic Diet. Fecal samples were taken and subsequently analyzed by real-time PCR. The results indicated different dietary patterns for subjects who consumed a higher amount of fruits, vegetables, legumes, and fish and a lower amount of bakery foods and precooked foods and snacks compared to Spanish consumption data. Most participants showed intermediate or high adherence to Southern European Atlantic Diet, and an analysis of gut microbiota showed high numbers of total bacteria and Actinobacteria, as well as high amounts of bacteria belonging to the genera Lactobacillus spp. and Bifidobacterium spp. A subsequent statistical comparison also revealed differences in gut microbiota depending on the subject's body weight, age, or degree of adherence to the Southern European Atlantic Diet

Food additives, contaminants and other minor components: effects on human gut microbiota-a review.

Gut bacteria play an important role in several metabolic processes and human diseases, such as obesity and accompanying co-morbidities, such as fatty liver disease, insulin resistance/diabetes, and cardiovascular events. Among other factors, dietary patterns, probiotics, prebiotics, synbiotics, antibiotics, and non-dietary factors, such as stress, age, exercise, and climatic conditions, can dramatically impact the human gut microbiota equilibrium and diversity. However, the effect of minor food constituents, including food additives and trace contaminants, on human gut microbiota has received less attention. Consequently, the present review aimed to provide an objective perspective of the current knowledge regarding the impacts of minor food constituents on human gut microbiota and consequently, on human health.

Influence of food consumption patterns and Galician lifestyle on human gut microbiota.

The proportion of different microbial populations in the human gut is an important factor that in recent years has been linked to obesity and numerous metabolic diseases. Because there are many factors that can affect the composition of human gut microbiota, it is of interest to have information about what is the composition of the gut microbiota in different populations in order to better understand the possibilities for improving nutritional management. A group of 31 volunteers were selected according to established inclusion and exclusion criteria and were asked about their diet history, lifestyle patterns, and adherence to the Southern European Atlantic Diet. Fecal samples were taken and subsequently analyzed by real-time PCR. The results indicated different dietary patterns for subjects who consumed a higher amount of fruits, vegetables, legumes, and fish and a lower amount of bakery foods and precooked foods and snacks compared to Spanish consumption data. Most participants showed intermediate or high adherence to Southern European Atlantic Diet, and an analysis of gut microbiota showed high numbers of total bacteria and Actinobacteria, as well as high amounts of bacteria belonging to the genera Lactobacillus spp. and Bifidobacterium spp. A subsequent statistical comparison also revealed differences in gut microbiota depending on the subject's body weight, age, or degree of adherence to the Southern European Atlantic Diet.

Exogenous FABP4 increases breast cancer cell proliferation and activates the expression of fatty acid transport proteins.

Adipose tissue plays an important role in tumor progression, because it provides nutrients and adipokines to proliferating cells. Fatty acid binding protein 4 (FABP4) is a key adipokine for fatty acid transport. In metabolic pathologies, plasma levels of FABP4 are increased. However, the role of this circulating protein is unknown. Recent studies have demonstrated that FABP4 might have a role in tumor progression, but the molecular mechanisms involved are still unclear. In this study, we analysed the role of eFABP4 (exogenous FABP4) in breast cancer progression. MCF-7 and MDA-MB-231 breast cancer cells did not express substantial levels of FABP4 protein, but intracellular FABP4 levels increased after eFABP4 incubation. Moreover, eFABP4 enhanced the proliferation of these breast cancer cells but did not have any effect on MCF-7 and MDA-MB-231 cell migration. Additionally, eFABP4 induced the AKT and MAPK signaling cascades in breast cancer cells, and the inhibition of these pathways reduced the eFBAP4-mediated cell proliferation. Interestingly, eFABP4 treatment in MCF-7 cells increased levels of the transcription factor FoxM1 and the fatty acid transport proteins CD36 and FABP5. In summary, we showed that eFABP4 plays a key role in tumor proliferation and activates the expression of fatty acid transport proteins in MCF-7 breast cancer cells. © 2016 Wiley Periodicals, Inc.

A Confirmatory Method Based on HPLC-MS/MS for the Detection and Quantification of Residue of Tetracyclines in Nonmedicated Feed.

The Commission Regulation 574/2011/EC set up maximum levels of coccidiostats and histomonostats in nonmedicated feed as a consequence of carry-over during manufacturing. Carry-over takes place from medicated to nonmedicated feed during feed production. Similar contamination could also occur for other pharmaceuticals such as tetracyclines, a group of antibiotics commonly employed in food production animal. The objective of this work is to present a simple and fast method for the simultaneous detection of four tetracyclines (chlortetracycline, doxycycline, oxytetracycline, and tetracycline) in nontarget feed at a µg/kg level. Validation of the method was performed according to the guideline included in the Commission Decision 2002/657/EC for official method. The validated method was successfully applied to 50 feed samples collected from different milk farms and 25 samples obtained from feed manufacturers. While oxytetracycline was the tetracycline most frequently detected, chlortetracycline was the analyte measured at the highest concentration 15.14 mg/Kg. From 75 nonmedicated feed analysed 15% resulted to be positive for the presence of one tetracycline.

Exogenous FABP4 induces endoplasmic reticulum stress in HepG2 liver cells.

BACKGROUND AND AIMS: Fatty acid binding protein 4 (FABP4) is an intracellular fatty acid (FA) carrier protein that is, in part, secreted into circulation. Circulating FABP4 levels are increased in obesity, diabetes and other insulin resistance (IR) diseases. FAs contribute to IR by promoting endoplasmic reticulum stress (ER stress) and altering the insulin signaling pathway. The effect of FABP4 on ER stress in the liver is not known. The aim of this study was to investigate whether exogenous FABP4 (eFABP4) is involved in the lipid-induced ER stress in the liver. METHODS: HepG2 cells were cultured with eFABP4 (40 ng/ml) with or without linoleic acid (LA, 200 µM) for 18 h. The expression of ER stress-related markers was determined by Western blotting (ATF6, EIF2α, IRE1 and ubiquitin) and real-time PCR (ATF6, CHOP, EIF2α and IRE1). Apoptosis was studied by flow cytometry using Annexin V-FITC and propidium iodide staining. RESULTS: eFABP4 increased the ER stress markers ATF6 and IRE1 in HepG2 cells. This effect led to insulin resistance mediated by changes in AKT and JNK phosphorylation. Furthermore, eFABP4 significantly induced both apoptosis, as assessed by flow cytometry, and CHOP expression, without affecting necrosis and ubiquitination. The presence of LA increased the ER stress response induced by eFABP4. CONCLUSIONS: eFABP4, per se, induces ER stress and potentiates the effect of LA in HepG2 cells, suggesting that FABP4 could be a link between obesity-associated metabolic abnormalities and hepatic IR mechanisms.

New insights into circulating FABP4: Interaction with cytokeratin 1 on endothelial cell membranes.

Fatty acid-binding protein 4 (FABP4) is an adipose tissue-secreted adipokine that is involved in the regulation of energetic metabolism and inflammation. Increased levels of circulating FABP4 have been detected in individuals with cardiovascular risk factors. Recent studies have demonstrated that FABP4 has a direct effect on peripheral tissues, specifically promoting vascular dysfunction; however, its mechanism of action is unknown. The objective of this work was to assess the specific interactions between exogenous FABP4 and the plasma membranes of endothelial cells. Immunofluorescence assays showed that exogenous FABP4 localized along the plasma membranes of human umbilical vein endothelial cells (HUVECs), interacting specifically with plasma membrane proteins. Anti-FABP4 immunoblotting revealed two covalent protein complexes containing FABP4 and its putative receptor; these complexes were approximately 108 kDa and 77 kDa in size. Proteomics and mass spectrometry experiments revealed that cytokeratin 1 (CK1) was the FABP4-binding protein. An anti-CK1 immunoblot confirmed the presence of CK1. FABP4-CK1 complexes were also detected in HAECs, HCASMCs, HepG2 cells and THP-1 cells. Pharmacological FABP4 inhibition by BMS309403 results in a slight decrease in the formation of these complexes, indicating that fatty acids may play a role in FABP4 functionality. In addition, we demonstrated that exogenous FABP4 crosses the plasma membrane to enter the cytoplasm and nucleus in HUVECs. These findings indicate that exogenous FABP4 interacts with plasma membrane proteins, specifically CK1. These data contribute to our current knowledge regarding the mechanism of action of circulating FABP4.

Interacción de FABP4 con proteínas de membrana de células endoteliales / Interaction of FABP4 with plasma membrane proteins of endothelial cells

Introducción: Fatty acid-binding protein 4 (FABP4) es una adipocina secretada por el tejido adiposo implicada en la regulación del metabolismo energético y la inflamación. FABP4 circulante se asocia con obesidad, dislipidemia aterogénica y síndrome metabólico. Estudios recientes muestran una asociación entre FABP4 circulante y disfunción endotelial, aunque se desconoce cómo se produce esta. El objetivo de este trabajo es estudiar la interacción entre FABP4 con las proteínas de la membrana citoplasmática en células endoteliales. Metodología: Se incubaron células HUVEC con y sin FABP4 (100 ng/ml) durante 5 min. La inmunolocalización de FABP4 se estudió mediante microscopia confocal. Para estudiar las interacciones de FABP4 con las proteínas de membrana de las células HUVEC se diseñó una estrategia que combina incubaciones con o sin 6XHistidine-tag FABP4 (FABP4-His) (100 ng/ml), cross-linking con formaldehído, extracción de proteínas de membrana y western blot. Resultados: Los resultados muestran que FABP4 colocaliza con CD31, una proteína utilizada como marcador de membrana citoplasmática. Además se observan diferentes patrones de western blot en función de la incubación con o sin FABP4-His. El inmunoblot revela la existencia de 3 complejos proteicos de aproximadamente 108, 77 y 33 kDa formados por FABP4 exógena y su posible receptor/es. Discusión: Los resultados obtenidos apoyan la existencia de un complejo proteico capaz de unir FABP4 a las células endoteliales mediante una unión específica. Además, nos permiten avanzar en el conocimiento de los efectos moleculares de FABP4 y, en caso de confirmarse, podrían utilizarse como diana terapéutica para prevenir enfermedades cardiovasculares

Introduction: Fatty acid binding protein (FABP4) is an adipose tissue-secreted adipokine implicated in the regulation of the energetic metabolism and inflammation. High levels of circulating FABP4 have been described in people with obesity, atherogenic dyslipidemia, diabetes and metabolic syndrome. Recent studies have demonstrated that FABP4 could have a direct effect on peripheral tissues and, specifically, on vascular function. It is still unknown how the interaction between FABP4 and the endothelial cells is produced to prompt these effects on vascular function. The objective of this work is studying the interaction between FABP4 and the plasma membrane proteins of endothelial cells. Methodology: HUVEC cells were incubated with and without FABP4 (100 ng/ml) for 5 minutes. Immunolocalization of FABP4 was studied by confocal microscopy. The results showed that FABP4 colocalizates with CD31, a membrane protein marker. A strategy which combines 6XHistidine-tag FABP4 (FABP4-His), incubations with or without FABP4-His (100 ng/ml), formaldehyde cross-linking, cellular membrane protein extraction and western blot, was designed to study the FABP4 interactions with membrane proteins of HUVECs. Results: The results showed different western blot profiles depending of the incubation with or without FABP4-His. The immunoblot revelead three covalent protein complexes of about 108, 77 and 33 kDa containing FAPB4 and its putative receptor. Discussion: The existence of a specific binding protein complex able to bind FABP4 to endothelial cells is supported by these results. The obtained results will permit us advance in the molecular knowledge of FABP4 effects as well as use this protein and its receptor as therapeutic target to prevent cardiovascular

Egg and egg-derived foods: effects on human health and use as functional foods.

Eggs are sources of protein, fats and micronutrients that play an important role in basic nutrition. However, eggs are traditionally associated with adverse factors in human health, mainly due to their cholesterol content. Nowadays, however, it is known that the response of cholesterol in human serum levels to dietary cholesterol consumption depends on several factors, such as ethnicity, genetic makeup, hormonal factors and the nutritional status of the consumer. Additionally, in recent decades, there has been an increasing demand for functional foods, which is expected to continue to increase in the future, owing to their capacity to decrease the risks of some diseases and socio-demographic factors such as the increase in life expectancy. This work offers a brief overview of the advantages and disadvantages of egg consumption and the potential market of functional eggs, and it explores the possibilities of the development of functional eggs by technological methods.

[Interaction of FABP4 with plasma membrane proteins of endothelial cells]. / Interacción de FABP4 con proteínas de membrana de células endoteliales.

INTRODUCTION: Fatty acid binding protein (FABP4) is an adipose tissue-secreted adipokine implicated in the regulation of the energetic metabolism and inflammation. High levels of circulating FABP4 have been described in people with obesity, atherogenic dyslipidemia, diabetes and metabolic syndrome. Recent studies have demonstrated that FABP4 could have a direct effect on peripheral tissues and, specifically, on vascular function. It is still unknown how the interaction between FABP4 and the endothelial cells is produced to prompt these effects on vascular function. The objective of this work is studying the interaction between FABP4 and the plasma membrane proteins of endothelial cells. METHODOLOGY: HUVEC cells were incubated with and without FABP4 (100 ng/ml) for 5 minutes. Immunolocalization of FABP4 was studied by confocal microscopy. The results showed that FABP4 colocalizates with CD31, a membrane protein marker. A strategy which combines 6XHistidine-tag FABP4 (FABP4-His), incubations with or without FABP4-His (100 ng/ml), formaldehyde cross-linking, cellular membrane protein extraction and western blot, was designed to study the FABP4 interactions with membrane proteins of HUVECs. RESULTS: The results showed different western blot profiles depending of the incubation with or without FABP4-His. The immunoblot revelead three covalent protein complexes of about 108, 77 and 33 kDa containing FAPB4 and its putative receptor. DISCUSSION: The existence of a specific binding protein complex able to bind FABP4 to endothelial cells is supported by these results. The obtained results will permit us advance in the molecular knowledge of FABP4 effects as well as use this protein and its receptor as therapeutic target to prevent cardiovascular.

FABP4 induces vascular smooth muscle cell proliferation and migration through a MAPK-dependent pathway.

PURPOSE: The migration and proliferation of vascular smooth muscle cells play crucial roles in the development of atherosclerotic lesions. This study examined the effects of fatty acid binding protein 4 (FABP4), an adipokine that is associated with cardiovascular risk, endothelial dysfunction and proinflammatory effects, on the migration and proliferation of human coronary artery smooth muscle cells (HCASMCs). METHODS AND RESULTS: A DNA 5-bromo-2'-deoxy-uridine (BrdU) incorporation assay indicated that FABP4 significantly induced the dose-dependent proliferation of HCASMCs with a maximum stimulatory effect at 120 ng/ml (13% vs. unstimulated cells, p<0.05). An anti-FABP4 antibody (40 ng/ml) significantly inhibited the induced cell proliferation, demonstrating the specificity of the FABP4 proliferative effect. FABP4 significantly induced HCASMC migration in a dose-dependent manner with an initial effect at 60 ng/ml (12% vs. unstimulated cells, p<0.05). Time-course studies demonstrated that FABP4 significantly increased cell migration compared with unstimulated cells from 4 h (23%vs. 17%, p<0.05) to 12 h (74%vs. 59%, p<0.05). Pretreatment with LY-294002 (5 µM) and PD98059 (10 µM) blocked the FABP4-induced proliferation and migration of HCASMCs, suggesting the activation of a kinase pathway. On a molecular level, we observed an up-regulation of the MAPK pathway without activation of Akt. We found that FABP4 induced the active forms of the nuclear transcription factors c-jun and c-myc, which are regulated by MAPK cascades, and increased the expression of the downstream genes cyclin D1 and MMP2, CCL2, and fibulin 4 and 5, which are involved in cell cycle regulation and cell migration. CONCLUSIONS: These findings indicate a direct effect of FABP4 on the migration and proliferation of HCASMCs, suggesting a role for this adipokine in vascular remodelling. Taken together, these results demonstrate that the FABP4-induced DNA synthesis and cell migration are mediated primarily through a MAPK-dependent pathway that activates the transcription factors c-jun and c-myc in HCASMCs.

Recovering Streptococcus mutans over restorations by the tray technique: a randomized clinical study

AIM: To correlate the tray technique for isolation and counting of Streptococcus mutans over dental restoration with the toothpick technique. METHODS: Forty 18-50-year-old patients of both genders with high cariogenic risk were selected. Inclusion criteria were: occlusal restoration evaluated as alpha, according to the Ryge's criteria (examiner' Cohen-Kappa 0.8) and 2 years old maximum age. Each patient provided a saliva sample (control group S) and two samples of dental biofilm from amalgam or resin-based composite restorations in the same patient, one obtained with the toothpick technique (group TT), and the other from homologous tooth restoration with the tray technique (group TR). This method involves only a direct impression on the occlusal surfaces by a section of a fluoride tray with trypticase yeast extract cysteine sucrose with bacitracin (TYCSB) agar previously prepared and solidified. The samples were incubated at 37 ºC for 48 h and the colony-forming units (CFU) were counted. The results were statistically analyzed with SPSS software using the linear regression method at 95% confidence level. RESULTS: TR was able to isolate S. mutans, and there was a significant positive correlation (r=0.95) with TT. There was no significant correlation between TR and control group S (r=0.47). CONCLUSIONS: TR was able to isolate and count S. mutans from the dental biofilm of dental restorations with a significant correlation with the TT group.

Fatty acid-binding protein 4 impairs the insulin-dependent nitric oxide pathway in vascular endothelial cells.

BACKGROUND: Recent studies have shown that fatty acid-binding protein 4 (FABP4) plasma levels are associated with impaired endothelial function in type 2 diabetes (T2D). In this work, we analysed the effect of FABP4 on the insulin-mediated nitric oxide (NO) production by endothelial cells in vitro. METHODS: In human umbilical vascular endothelial cells (HUVECs), we measured the effects of FABP4 on the insulin-mediated endothelial nitric oxide synthase (eNOS) expression and activation and on NO production. We also explored the impact of exogenous FABP4 on the insulin-signalling pathway (insulin receptor substrate 1 (IRS1) and Akt). RESULTS: We found that eNOS expression and activation and NO production are significantly inhibited by exogenous FABP4 in HUVECs. FABP4 induced an alteration of the insulin-mediated eNOS pathway by inhibiting IRS1 and Akt activation. These results suggest that FABP4 induces endothelial dysfunction by inhibiting the activation of the insulin-signalling pathway resulting in decreased eNOS activation and NO production. CONCLUSION: These findings provide a mechanistic linkage between FABP4 and impaired endothelial function in diabetes, which leads to an increased cardiovascular risk.