Retinoic Acid Increases Fatty Acid Oxidation and Irisin Expression in Skeletal Muscle Cells and Impacts Irisin In Vivo.

BACKGROUND/AIMS: All-trans retinoic acid (ATRA) has protective effects against obesity and metabolic syndrome. We here aimed to gain further insight into the interaction of ATRA with skeletal muscle metabolism and secretory activity as important players in metabolic health. METHODS: Cultured murine C2C12 myocytes were used to study direct effects of ATRA on cellular fatty acid oxidation (FAO) rate (using radioactively-labelled palmitate), glucose uptake (using radioactively-labelled 2-deoxy-D-glucose), triacylglycerol levels (by an enzymatic method), and the expression of genes related to FAO and glucose utilization (by RT-real time PCR). We also studied selected myokine production (using ELISA and immunohistochemistry) in ATRA-treated myocytes and intact mice. RESULTS: Exposure of C2C12 myocytes to ATRA led to increased fatty acid consumption and decreased cellular triacylglycerol levels without affecting glucose uptake, and induced the expression of the myokine irisin at the mRNA and secreted protein level in a dose-response manner. ATRA stimulatory effects on FAO-related genes and the Fndc5 gene (encoding irisin) were reproduced by agonists of peroxisome proliferator-activated receptor ß/δ and retinoid X receptors, but not of retinoic acid receptors, and were partially blocked by an AMP-dependent protein kinase inhibitor. Circulating irisin levels were increased by 5-fold in ATRA-treated mice, linked to increased Fndc5 transcription in liver and adipose tissues, rather than skeletal muscle. Immunohistochemistry analysis of FNDC5 suggested that ATRA treatment enhances the release of FNDC5/irisin from skeletal muscle and the liver and its accumulation in interscapular brown and inguinal white adipose depots. CONCLUSION: These results provide new mechanistic insights on how ATRA globally stimulates FAO and enhances irisin secretion, thereby contributing to leaning effects and improved metabolic status.

Synergistic Effects of a Mixture of Glycosaminoglycans to Inhibit Adipogenesis and Enhance Chondrocyte Features in Multipotent Cells.

BACKGROUND/AIMS: Multipotent mesenchymal stem cells affect homeostasis of adipose and joint tissues. Factors influencing their differentiation fate are of interest for both obesity and joint problems. We studied the impact of a mixture of glycosaminoglycans (GAGs) (hyaluronic acid: dermatan sulfate 1:0.25, w/w) used in an oral supplement for joint discomfort (Oralvisc™) on the differentiation fate of multipotent cells. METHODS: Primary mouse embryo fibroblasts (MEFs) were used as a model system. Post-confluent monolayer MEF cultures non-stimulated or hormonally stimulated to adipogenesis were chronically exposed to the GAGs mixture, its individual components or vehicle. The appearance of lipid laden cells, lipid accumulation and expression of selected genes at the mRNA and protein level was assessed. RESULTS: Exposure to the GAGs mixture synergistically suppressed spontaneous adipogenesis and induced the expression of cartilage extracellular matrix proteins, aggrecan core protein, decorin and cartilage oligomeric matrix protein. Hormonally-induced adipogenesis in the presence of the GAGs mixture resulted in decreased adipogenic differentiation, down-regulation of adipogenic/lipogenic factors and genes for insulin resistance-related adipokines (resistin and retinol binding protein 4), and up-regulation of oxidative metabolism-related genes. Adipogenesis in the presence of dermatan sulfate, the minor component of the mixture, was not impaired but resulted in smaller lipid droplets and the induction of a more complete brown adipocyte-related transcriptional program in the cells in the adipose state. CONCLUSIONS: The Oralvisc™ GAGs mixture can tip the adipogenic/chondrogenic fate balance of multipotent cells away from adipogenesis while favoring chondrocyte related gene expression. The mixture and its dermatan sulfate component also have modulatory effects of interest on hormonally-induced adipogenesis and on metabolic and secretory capabilities of adipose cells.

ß-Carotene during the suckling period is absorbed intact and induces retinoic acid dependent responses similar to preformed vitamin A in intestine and liver, but not adipose tissue of young rats.

SCOPE: We studied ß-carotene (BC) absorption and metabolism and compared BC and retinyl palmitate (RE) for their impact on white adipose tissue (WAT) development in suckling rats. METHODS AND RESULTS: Rat pups received daily orally from days 1-20 of life either the vehicle or vitamin A (approx. ×3 that ingested daily from maternal milk) in the form of BC or RE. Intact BC was found in serum and liver of BC-supplemented rats. Both BC and RE supplementation increased retinoic acid mediated transcriptional responses in intestine (on Isx and Bco1) and the liver (on Cyp26a1 and Cpt1a). In contrast, responses in WAT were dependent on the vitamin A source: WAT of BC-supplemented rats, like WAT of control rats, was enriched in larger adipocytes with increased adipogenic markers (peroxisome proliferator-activated receptor γ and downstream genes) and reduced markers of proliferative status (proliferating cell nuclear antigen) compared to WAT of RE-supplemented rats. CONCLUSION: BC is partly absorbed intact by suckling rats, which resembles the situation in humans and suggests that suckling rats may be an appropriate animal model to study BC uptake, metabolism and biological activity, particularly in infants. Vitamin A supplementation with BC or RE in early life differentially affects WAT and may thus entail different outcomes regarding adiposity programming.

Molecular players at the intersection of obesity and osteoarthritis.

Obesity and degenerative joint disease (osteoarthritis, OA) are two multifactorial pathologies that are becoming major medical issues with the aging of the world population. The relationship of OA with obesity is complex, involving both biomechanical and metabolic links. Dysregulated production of adipose tissue-derived inflammatory mediators, hyperlipidemia, and increased systemic oxidative stress are conditions frequently associated with obesity that may favor joint degeneration. In addition, it is remarkable that many regulatory factors have been implicated in the development, maintenance and function of both adipose tissues and cartilage and other articular joint tissues. Disturbances in these factors may underlie additional links between obesity and OA. In this review, molecular players at the intersection of adipose tissue and joint cell biology - including differentiation signals and transcription factors, extracellular matrix components and remodelers, joint cell- and adipose tissue cell-derived mediators (cytokines, adipokines), hypoxia inducible transcription factors, lipids, advanced glycation end products and miRNAs - are reviewed, with emphasis on their dysregulation in obesity and OA. Knowledge of these factors may illuminate a novel, adipocentric avenue for the pathogenesis and therapy of OA and other joint diseases.

Distinct effects of oleic acid and its trans-isomer elaidic acid on the expression of myokines and adipokines in cell models.

Trans-fatty acids (TFA) and cis-monounsaturated fat appear to exert detrimental and beneficial effects, respectively, on glucose metabolism and insulin sensitivity. Adipose tissue and skeletal muscle are a source of signalling proteins (adipokines and myokines), some of which have been related to the control of insulin sensitivity. Here, we investigated the possible differential effects of elaidic acid (EA; trans-9-18 : 1) - the major component in industrially produced TFA - and oleic acid (OA; cis-9-18 : 1) - its cis-isomer naturally present in food - on cellular glucose uptake and the expression of selected myokines and adipokines using cell models. Differentiated C2C12 myotubes and 3T3-L1 adipocytes were pretreated with the vehicle (control cells) or fatty acids for 24 h, after which basal and insulin-stimulated 2-deoxyglucose uptake and the expression of selected signalling proteins were measured. In C2C12 myotubes, pretreatment with OA, but not with EA, led to increased insulin-stimulated 2-deoxyglucose uptake and IL-6 expression levels, while pretreatment with EA, but not with OA, led to reduced IL-15 mRNA levels and increased TNF-α expression levels. In 3T3-L1 adipocytes, exposure to OA, but not to EA, resulted in reduced resistin gene expression and increased adiponectin gene expression. The results show evidence of distinct, direct effects of OA and EA on muscle glucose uptake and the expression of target myokines and adipokines, thus suggesting novel mechanisms by which cis- and trans-monounsaturated fat may differentially affect systemic functions.

Unusual case of osteosynthesis failure in a hip fracture: side-plate pull-out.

Osteoporotic hip fractures are common in our setting. Poor bone quality favors complications of the osteosynthesis procedures used to treat these patients. Lag-screw cut-out through the femoral head is not uncommon (2%), but pull-out of side plate screws is very unusual. We present the case of a patient with a stable osteoporotic fracture treated by osteosynthesis using a four-hole plate, who presented with a pull-out following a low-energy fall.

Fracture of the unresurfaced patella after total knee arthroplasty: a report of two cases.

Patellar fractures are unusual in total knee arthroplasty without patellar resurfacing. We present 2 such cases that occurred within postoperative 2 months and were managed conservatively. Both patients had their knee function preserved.

All-trans retinoic acid decreases murine adipose retinol binding protein 4 production.

BACKGROUND/AIMS: Adipose-derived retinol binding protein 4 (RBP4) might contribute to the development of insulin resistance, and therefore further knowledge of factors regulating it is of interest. Retinoic acid, the acid form of vitamin A, affects the expression of several adipokines related to insulin sensitivity in mice. Here, we sought to investigate its impact on adipose RBP4 production. METHODS: Changes in RBP4 expression were analyzed in adipose tissues and liver of mice treated in vivo with all-trans retinoic acid (ATRA), and in 3T3-L1 adipocytes and adipocytes derived from mouse embryonic fibroblasts exposed to ATRA. RESULTS: ATRA treatment in mice increased insulin sensitivity as assessed by the homeostatic model assessment for insulin resistance, and led to a reduction of RBP4 mRNA and protein levels in adipose tissues, a reduction of RBP4 protein but not RBP4 mRNA levels in the liver, and a marked increase in circulating RBP4 protein levels. In adipocyte cell models, ATRA down-regulated RBP4 mRNA levels in a dose-dependent manner: this effect was reproduced by retinaldehyde and retinoid receptors agonists, and correlated with a reduced accumulation of RBP4 protein in the culture medium. CONCLUSION: These results reveal a selective effect of ATRA inhibiting RBP4 expression specifically in adipocytes, and reinforce the concept that vitamin A vitamers may affect insulin sensitivity through effects on adipokine production.

[Clinical implications and prognostic significance of the study on the circadian variation of heart rate variability in patients with severe pulmonary hypertension]. / Implicaciones clínicas y pronósticas del estudio circadiano de la modulación simpático-vagal de la variabilidad de la frecuencia cardiaca en pacientes con hipertensión arterial pulmonar grave.

BACKGROUND: A reduction of heart rate variability (HRV) is currently considered an independent risk factor for morbidity, mortality and severity of severalcardiac disease, however, the dynamic sympathovagal modulation on HRV during 24 hr in primary pulmonary hypertension (PPH) had not been described. METHODS: 24 hr Holter monitoring (HA) were recorded in 32 patients (mean age 34, +/-12, 90% female) with severe primary pulmonary hypertension (mean pulmonary pressure, 90:t:12 mm Hg), and in 34 patients (mean age 36 +/-14, 60% female) with Eisenmenger syndrome (ES) secondary to septal ventricular defect or atent ductus arteriosus. A control group (n=44) paired for age, gender and arterial pulmonary pressure was included. HRV time and spectral parameters (mean, SDNN, SDANN, rMSSD, PNN50, LF, HF and LF/HF ratio) were analyzed during three periods: 24 hr; day (8-22:00), night (23-07:00) and also every hour of recording at 5 min-intervals). After detection of sympatho-vagal balance 15 patients were randomized, Treprostinil (prostaglandin) was administered to 6 patients and subcutaneous placebo to 9. RESULTS: HRV frequency parameters during 24 hr HM were significantly different among groups. LF/HF (day) 5.9:1:12.5:1:1P.001 and LF/HF night) 2.8:tlvs.1.5:l:.8.034. Sympathovagal modulation on 24 hr HRV showed that heart rate circadian rhythm is clearly altered in both PPH and ES, but the sympathetic tone in PPH is higher at l 24 hr. (p < .05), after administering treprostinil a recovery of sympathovagal balance was observed CONCLUSIONS: Autonomic cardiac disturbance is clearly present in PPH and ES. The circadian rhythm of HRV is first lost due to an increase of sympathetic tone. These changes may be markers of autonomic disbalance that favor the development of arrhythmias and sudden death. The sympathovagal balance in PPH could be considered an important risk marker.

Implicaciones clínicas y pronósticas del estudio circadiano de la modulación simpático-vagal de la variabilidad de la frecuencia cardiaca en pacientes con hipertensión arterial pulmonar grave / Clinical implications and prognostic significance of the study on the circadian variation of heart rate variability in patients with severe pulmonary hypertension

Introducción: La reducción en la variabilidad de la frecuencia cardiaca ha sido identificada como factor de riesgo en enfermedad cardiovascular, pero su descripción en hipertensión arterial pulmonar severa se desconoce. Material y métodos: Se estudiaron pacientes con hipertensión arterial pulmonar grave, 32 con hipertensión pulmonar primaria, 34 con hipertensión pulmonar secundaria a cardiopatía congénita (Eisenmenger) y 44 sujetos control sin evidencia de enfermedad. La evaluación del registro ambulatorio de la frecuencia cardiaca se realizó por métodos convencionales. El análisis espectral y la relación a baja y alta frecuencia se realizó utilizando el método de Fourier. Comparaciones entre día y noche se realizó entre los grupos. Después de conocer el perfil circadiano, 15 pacientes con hipertensión pulmonar fueron seleccionados para recibir tratamiento al azar con Treprostinil (Prostaglandina) o placebo por vía subcutánea. Posteriormente (3 meses) se analizaron nuevamente los parámetros de variabilidad de frecuencia cardiaca y de hemodinámica para conocer el impacto de dicha terapéutica. Resultados: Se detectó un estado franco de hipertonía simpática en el grupo de hipertensión pulmonar, sobre todo en los pacientes con hipertensión pulmonar primaria. El efecto de Treprostinil fue claramente asociado con disminución del tono simpático y un aumento de la capacidad física. Conclusiones: Los pacientes con hipertensión arterial pulmonar, cursan con equilibrio simpático-vagal alterado sobre todo durante el día. Hay pérdida del ritmo circadiano. Dichos trastornos pueden ser reversibles con la aplicación de treprostinil. El equilibrio simpáticovagal de la frecuencia cardiaca es un instrumento no invasivo que permite estratificar mejor al paciente con hipertensión arterial pulmonar grave.

BACKGROUND: A reduction of heart rate variability (HRV) is currently considered an independent risk factor for morbidity, mortality and severity of severalcardiac disease, however, the dynamic sympathovagal modulation on HRV during 24 hr in primary pulmonary hypertension (PPH) had not been described. METHODS: 24 hr Holter monitoring (HA) were recorded in 32 patients (mean age 34, +/-12, 90% female) with severe primary pulmonary hypertension (mean pulmonary pressure, 90:t:12 mm Hg), and in 34 patients (mean age 36 +/-14, 60% female) with Eisenmenger syndrome (ES) secondary to septal ventricular defect or atent ductus arteriosus. A control group (n=44) paired for age, gender and arterial pulmonary pressure was included. HRV time and spectral parameters (mean, SDNN, SDANN, rMSSD, PNN50, LF, HF and LF/HF ratio) were analyzed during three periods: 24 hr; day (8-22:00), night (23-07:00) and also every hour of recording at 5 min-intervals). After detection of sympatho-vagal balance 15 patients were randomized, Treprostinil (prostaglandin) was administered to 6 patients and subcutaneous placebo to 9. RESULTS: HRV frequency parameters during 24 hr HM were significantly different among groups. LF/HF (day) 5.9:1:12.5:1:1P.001 and LF/HF night) 2.8:tlvs.1.5:l:.8.034. Sympathovagal modulation on 24 hr HRV showed that heart rate circadian rhythm is clearly altered in both PPH and ES, but the sympathetic tone in PPH is higher at l 24 hr. (p < .05), after administering treprostinil a recovery of sympathovagal balance was observed CONCLUSIONS: Autonomic cardiac disturbance is clearly present in PPH and ES. The circadian rhythm of HRV is first lost due to an increase of sympathetic tone. These changes may be markers of autonomic disbalance that favor the development of arrhythmias and sudden death. The sympathovagal balance in PPH could be considered an important risk marker.

[Evaluation of coronary artery disease by myocardial perfusion imaging in women]. / Evaluación de enfermedad arterial coronaria con imágenes de perfusión miocárdica en mujeres.

UNLABELLED: The early diagnosis of coronary artery disease continues to be a challenge in women because non invasive diagnostic tests to evaluate ischemia, such as stress test are less accurate in this group of patients. The aim of the study was to assess the value of myocardial perfusion imaging with SPECT for the detection of ischemia. METHODS: We studied 151 consecutive women who were assessed by SPECT with Tc-99m Sestamibi, stress test and coronary angiography. Sensitivity and specificity was calculated for each test. RESULTS: The prevalence of angiographycally significant coronary artery disease was 71.5%. The sensitivity and specificity of myocardial perfusion imaging for the detection of ischemia was 91.6% and 87.9% respectively. The sensitivity and specificity of stress test was 43% and 96.1%. Accuracy of myocardial perfusion imaging was higher in the group of patients older than 50 years of age, in whom severe coronary artery disease was more prevalent. CONCLUSIONS: The result of this study suggest that myocardial perfusion imaging is a very useful tool for detection of ischemia in women with CAD, especially for those over 50 years of age, being superior to other non invasive tests as stress test.

Evaluación de enfermedad arterial coronaria con imágenes de perfusión miocárdica en mujeres / Evaluation of coronary artery disease by myocardial perfusion imaging in women

El diagnóstico precoz de la enfermedad arterial coronaria constituye un reto en las mujeres, en quienes los métodos diagnósticos no invasivos para evaluar isquemia tales como prueba de esfuerzo tienen menor precisión. El objetivo del estudio fue conocer la utilidad de las imágenes de perfusión miocárdica con SPECT para la evaluación de isquemia en mujeres. Métodos: Se estudiaron 151 mujeres a las cuales se les realizaron imágenes de perfusión miocárdica con SPECT y Tc-99m- Sestamibi, electrocardiografía de esfuerzo, y angiografía. Se evaluó la sensibilidad y especificidad de cada prueba. Resultados: La prevalencia de enfermedad arterial coronaria angiográficamente significativa fue de 71.5%. La sensibilidad y especificidad de las imágenes de SPECT para la detección de isquemia fue de 91.6% y 87.9%. La sensibilidad y especificidad de la electrocardiografía de esfuerzo fue de 43% y 96.1%. La precisión de las imágenes de SPECT es mayor en pacientes mayores de 50 años, en las cuales se encontró mayor prevalencia y severidad de la enfermedad arterial coronaria. Conclusiones: Las imágenes de perfusión miocárdica con SPECT son de gran utilidad para la detección de isquemia en mujeres con enfermedad arterial coronaria, sobre todo en mayores de 50 años, siendo superior a la electrocardiografía de esfuerzo.

The early diagnosis of coronary artery disease continues to be a challenge in women because non invasive diagnostic tests to evaluate ischemia, such as stress test are less accurate in this group of patients. The aim of the study was to assess the value of myocardial perfusion imaging with SPECT for the detection of ischemia. Methods: We studied 151 consecutive women who were assessed by SPECT with Tc-99m Sestamibi, stress test and coronary angiography. Sensitivity and specificity was calculated for each test. Results: The prevalence of angiographycally significant coronary artery disease was 71.5%. The sensitivity and specificity of myocardial perfusion imaging for the detection of ischemia was 91.6% and 87.9% respectively. The sensitivity and specificity of stress test was 43% and 96.1%. Accuracy of myocardial perfusion imaging was higher in the group of patients older than 50 years of age, in whom severe coronary artery disease was more prevalent. Conclusions: The result of this study suggest that myocardial perfusion imaging is a very useful tool for detection of ischemia in women with CAD, especially for those over 50 years of age, being superiorto other non invasive tests as stress test.

Atrial septal defect in adults > or =40 years old: negative impact of low arterial oxygen saturation.

BACKGROUND: Although good prognosis and clinical long-term outcome have been commonly reported in minimally symptomatic adult patients with ASD, this information has been based on studies with a relatively small number of adult patients. We studied unoperated patients aged over 40 years to define the patterns of presentation, anatomical characteristics, outcome and predictive factors for free-event survival of major cardiovascular and pulmonary events. METHODS AND RESULTS: Two-hundred survivors of atrial septal defect aged over 40-yr attended from 1985 to 1998 were reviewed and followed-up from 1.6 to 22 years. Patients were classified in three groups according to age at entry: Group 1, between 40 and 49; Group 2; 50 and 59; and Group 3, > or =60 years old. The mean age at presentation was 48.8+/-9.2 years, and the most common clinical presentations were arrhythmia and dyspnea (51.4%). There were 37 (18.5%) events: 7 heart failure-related, 5 sudden death, 13 severe pulmonary infections, 5 embolisms, and 4 strokes. According to Cox's regression analysis, predictors of primary end point included age group at presentation (hazard ratio 1.71, 95% confidence limits 1.16 to 2.54), and either pulmonary hypertension (mean pulmonary pressure >35 mmHg; hazard ratio=0.65 (4.6, confidence limits 2.2 to 9.5) or, arterial oxygen saturation <80% (hazard ratio 1.71, 95% confidence limits 1.16 to 2.54). CONCLUSIONS: This study supports that long term outcome of patients aged >40 years with unoperated ASD is importantly determined by the mPAP (>35 mmHg), SaO2% (_80) and the age at diagnosis. Nevertheless we identified an inverse association between the mPAP level and SaO2% (interaction). The event-free survival expectancy may be estimated using the age at diagnosis and either SaO2% or mPAP. This prognostic stratification based on pathophysiological principles, may help in making decisions for therapeutic interventions. SaO2% should always be measured as a part of the initial clinical approach of those patients with atrial septal defect aged over 40 years.

Anatomic three-dimensional echocardiographic correlation of bicuspid aortic valve.

This study was undertaken to verify the echocardiographic characteristics of bicuspid aortic valve (AV) using 3-dimensional transesophageal echocardiography by comparing the findings with anatomic examination of autopsy specimens from carriers of this condition. Three-dimensional reconstructions of transesophageal echocardiograms were performed on 14 patients with bicuspid AV, and 20 autopsy specimens of bicuspid AVs were analyzed. Echocardiographic images and autopsy material were correlated. Two variants of bicuspid aorta were identified. In group I the AV had 2 leaflets. This group included 9 (9/14) 3-dimensional echocardiographic studies and 13 (13/20) necropsies. In group II 3 sigmoid leaflets had originally developed and 2 underwent dysplastic fusion, resulting in functionally bicuspid valves. Five (5/14) echocardiographic studies and 7 (7/20) anatomic specimens fell into this category. There was a clear correspondence between anatomic and echocardiographic findings, which leads to the conclusion that 3-dimensional echocardiography is a technique that reliably defines the morphological details of bicuspid AV with the precision of anatomopathologic examination.