Predictive value of the advanced lipoprotein profile and glycated proteins on diabetic retinopathy

This study aimed to assess whether the advanced characteristics of serum lipoprotein subclasses could better predict the risk of developing diabetic retinopathy (DR) and its severity compared to other established risk factors in subjects with type 1 (T1D) and type 2 (T2D) diabetes. This observational, cross-sectional substudy analyzed DR-related data from 309 T1D and 264 T2D subjects. The advanced lipoprotein and glycoprotein profile was determined by nuclear magnetic resonance (NMR) spectroscopy (Liposcale test). NMR analysis of lipoproteins revealed that T1D subjects with DR showed standard non-HDL particles, despite higher IDL lipid concentrations. Notably, IDL lipids were elevated in T1D subjects with worsened DR. VLDL and LDL were smaller, whereas HDL triglycerides were increased in DR compared with non-DR. On the other hand, the T2D subjects with DR showed altered characteristics in the LDL fraction, mainly revealed by a significant decrease in smaller LDL and a reduction in LDL-C. Moreover, the glycoprotein profile did not reveal significant changes among DR groups, regardless of the type of diabetes. However, lipoprotein characteristics and glycoproteins unveiled by NMR analysis did not improve the predictive value of conventional lipids or other traditional, well-established biomarkers of DR in our cohorts

Anti-obesity sodium tungstate treatment triggers axonal and glial plasticity in hypothalamic feeding centers

Objective: This study aims at exploring the effects of sodium tungstate treatment on hypothalamic plasticity, which is known to have an important role in the control of energy metabolism. Methods: Adult lean and high-fat diet-induced obese mice were orally treated with sodium tungstate. Arcuate and paraventricular nuclei and lateral hypothalamus were separated and subjected to proteomic analysis by DIGE and mass spectrometry. Immunohistochemistry and in vivo magnetic resonance imaging were also performed. Results: Sodium tungstate treatment reduced body weight gain, food intake, and blood glucose and triglyceride levels. These effects were associated with transcriptional and functional changes in the hypothalamus. Proteomic analysis revealed that sodium tungstate modified the expression levels of proteins involved in cell morphology, axonal growth, and tissue remodeling, such as actin, CRMP2 and neurofilaments, and of proteins related to energy metabolism. Moreover, immunohistochemistry studies confirmed results for some targets and further revealed tungstate-dependent regulation of SNAP25 and HPC-1 proteins, suggesting an effect on synaptogenesis as well. Functional test for cell activity based on c-fos- positive cell counting also suggested that sodium tungstate modified hypothalamic basal activity. Finally, in vivo magnetic resonance imaging showed that tungstate treatment can affect neuronal organization in the hypothalamus. Conclusions: Altogether, these results suggest that sodium tungstate regulates proteins involved in axonal and glial plasticity. The fact that sodium tungstate could modulate hypothalamic plasticity and networks in adulthood makes it a possible and interesting therapeutic strategy not only for obesity management, but also for other neurodegenerative illnesses like Alzheimer's disease

Predictive Value of the Advanced Lipoprotein Profile and Glycated Proteins on Diabetic Retinopathy

This study aimed to assess whether the advanced characteristics of serum lipoprotein subclasses could better predict the risk of developing diabetic retinopathy (DR) and its severity compared to other established risk factors in subjects with type 1 (T1D) and type 2 (T2D) diabetes. This observational, cross-sectional substudy analyzed DR-related data from 309 T1D and 264 T2D subjects. The advanced lipoprotein and glycoprotein profile was determined by nuclear magnetic resonance (NMR) spectroscopy (Liposcale test). NMR analysis of lipoproteins revealed that T1D subjects with DR showed standard non-HDL particles, despite higher IDL lipid concentrations. Notably, IDL lipids were elevated in T1D subjects with worsened DR. VLDL and LDL were smaller, whereas HDL triglycerides were increased in DR compared with non-DR. On the other hand, the T2D subjects with DR showed altered characteristics in the LDL fraction, mainly revealed by a significant decrease in smaller LDL and a reduction in LDL-C. Moreover, the glycoprotein profile did not reveal significant changes among DR groups, regardless of the type of diabetes. However, lipoprotein characteristics and glycoproteins unveiled by NMR analysis did not improve the predictive value of conventional lipids or other traditional, well-established biomarkers of DR in our cohorts.This work was funded by the Ministerio de Sanidad y Consumo, Instituto de Salud Carlos III (Madrid, Spain), grant PI15/0625 (to D.M. and E.C.), PI17/01362 (to N.A.), and PI17/00232 (to J.J.), FEDER “Una manera de hacer Europa,” and by Fundació La Marató de TV3 2016 (303/C/2016) (201602.30.31) (to N.A. and J.J.). J.J. was a recipient of a Miguel Servet Type 2 contract (CPII18/00004; ISCIII). This research was supported by CIBER-Consorcio Centro de Investigación Biomédica en Red-CIBERDEM (CB15/00071), Instituto de Salud Carlos III, Ministerio de Ciencia e Innovación. Institut de Recerca de l’Hospital de la Santa Creu i Sant Pau is accredited by the Generalitat de Catalunya as Centre de Recerca de Catalunya (CERCA)

Predictive Value of the Advanced Lipoprotein Profile and Glycated Proteins on Diabetic Retinopathy

Fundació La Marató de TV3 2016This study aimed to assess whether the advanced characteristics of serum lipoprotein subclasses could better predict the risk of developing diabetic retinopathy (DR) and its severity compared to other established risk factors in subjects with type 1 (T1D) and type 2 (T2D) diabetes. This observational, cross-sectional substudy analyzed DR-related data from 309 T1D and 264 T2D subjects. The advanced lipoprotein and glycoprotein profile was determined by nuclear magnetic resonance (NMR) spectroscopy (Liposcale test). NMR analysis of lipoproteins revealed that T1D subjects with DR showed standard non-HDL particles, despite higher IDL lipid concentrations. Notably, IDL lipids were elevated in T1D subjects with worsened DR. VLDL and LDL were smaller, whereas HDL triglycerides were increased in DR compared with non-DR. On the other hand, the T2D subjects with DR showed altered characteristics in the LDL fraction, mainly revealed by a significant decrease in smaller LDL and a reduction in LDL-C. Moreover, the glycoprotein profile did not reveal significant changes among DR groups, regardless of the type of diabetes. However, lipoprotein characteristics and glycoproteins unveiled by NMR analysis did not improve the predictive value of conventional lipids or other traditional, well-established biomarkers of DR in our cohorts

Efecte Hipotalàmic del tungstat de sodi i la seva relació amb la vida de la leptina: una possible teràpia antiobesitat

[cat] L'obesitat s'ha convertit en un problema sanitari important a nivell mundial, i és important poder-hi trobar solucions. El tungstat, tal i com han demostrat els diversos estudis desenvolupats en el nostre laboratori, és un fàrmac amb un gran potencial en el tractament de l'obesitat. Tot i haver-se descrit àmpliament els diversos efectes del tungstat en diferents models experimentals, el mecanisme d'acció n'és encara desconegut. Els resultats preliminars indiquen que la leptina i la seva via de senyalització juguen un paper important en les accions del tungstat sobre l'homeòstasi energètica. Donat que l'hipotàlem és una de les dianes més importants de la leptina i que és des d'aquí on aquesta hormona regula el balanç energètic, la nostra hipòtesi de treball és que el tungstat desenvolupa part de les seves accions a aquest nivell per poder regular el guany de pes corporal i la ingesta dels animals tractats. Amb la intenció de veure si el tungstat tenia efectes centrals directes, es va administrar una injecció via intraperitoneal de tungstat de sodi a rates wistar, i se'n van mesurar els nivells detectables a líquid céfaloraquidi mitjançant espectrometria de masses. Els resultats van mostrar que el tungstat passa la barrera hematoencefàlica. El segon punt va ser veure que quan el tungstat s'administra per via central (injecció intracerebroventricular al tercer ventricle de l'hipotàlem), dóna lloc a la reducció del guany de pes i la ingesta dels animals tractats, i també incrementa els nivells de fosforilació de les principals proteïnes de la via de la leptina: JAK2, AKT i ERK1/2, mostrant que el tungstat té efectes centrals directes. In vitro, es va tractar la línia cel·lular hipotalàmica N29 amb tungstat 100µM o leptina 1nM. El tungstat va incrementar la fosforilació de les proteïnes JAK2, ERK1/2 i AKT, però l'activació de cada una d'aquestes cinases no depenia de cap de les altres, mostrant que els seus efectes són independents. A més, el tractament amb tungstat regula l'expressió gènica de c-myc a través de la seva acció sobre la via JAK/STAT, i l'expressió de c-fos i Agrp a través de l'acció sobre la via ERK1/2, amb dos efectes altre cop independents. Es van realitzar estudis de proteòmica, comparant els nuclis hipotalàmics ARC, PVN i LHA entre animals prims i obesos, tractats i no tractats amb tungstat per via oral. Els resultats van mostrar que el tractament amb aquesta molècula altera l'expressió de diferents proteïnes, moltes d'elles involucrades amb la plasticitat neuronal i el creixement axonal, suggerint un rol important de la molècula en aquestes accions. Tenint en compte que en l'obesitat, les connexions neuronals entre els diferents nuclis hipotalàmics són defectives, aquest resultat és molt prometedor. Tots els resultats exposats suggereixen que el tungstat de sodi actua, en alguns aspectes, com un mimètic de la leptina a nivell neuronal, modulant la via de transducció del senyal de l'hormona i possiblement, modulant la plasticitat neuronal. Aquests efectes obririen noves vies pel tractament de l'obesitat.[eng] Sodium tungstate is an antiobesity drug targeting peripheral tissues, but also modulates hypothalamic gene expression when orally administered, raising the possibility of a direct effect of sodium tungstate in the central nervous system. To assess whether sodium tungstate could have direct central effects, first, we administered a single intraperitoneal injection of sodium tungstate to wistar rats and its levels were measured in cerebrospinal fluid through mass spectrometry. Sodium tungstate crossed the blood brain barrier, reaching a concentration of 1.31±0.07 mg/l in cerebrospinal fluid 30 min after ip injection. Second, when centrally administered (a single intracerebroventricular injection into the third ventricle) sodium tungstate decreased body weight gain and food intake and increased the phosphorylation state of the main kinases and proteins involved in leptin signalling. In vitro, N29/4 neural cells were treated with 100µM sodium tungstate or 1nM leptin. Sodium tungstate increased the phosphorylation of janus kinase-2 (JAK2), extracellular signal-regulated kinase-1/2 (ERK1/2) and protein kinase-B (AKT), but the activation of each kinase did not depend on each other. Thus, it regulated c-myc gene expression through the JAK2/STAT system and c-fos and agrp gene expression through the ERK1/2 pathway simultaneously and independently. Proteomic studies were also performed, comparing the ARC, PVN and LHA hypothalamic nuclei from lean, obese and treated with sodium tungstate (orally) or not treated animals. The results show that treatment alters many different proteins, some of them involved in neuronal plasticity and axonal growth. All these data demonstrate that sodium tungstate had direct neural effects activating the leptin signalling pathway and increasing the activity of several kinases involved in the leptin signalling system in an independent way. Sodium tungstate also alters the expression of different proteins involved in neuronal plasticity, making it a suitable candidate to repair the neuronal connections altered because of the obesity. All these results present sodium tungstate as a suitable and promising candidate for improving central leptin sensitivity and managing obesity

Anti-obesity sodium tungstate treatment triggers axonal and glial plasticity in hypothalamic feeding centers.

International audienceOBJECTIVE: This study aims at exploring the effects of sodium tungstate treatment on hypothalamic plasticity, which is known to have an important role in the control of energy metabolism. METHODS: Adult lean and high-fat diet-induced obese mice were orally treated with sodium tungstate. Arcuate and paraventricular nuclei and lateral hypothalamus were separated and subjected to proteomic analysis by DIGE and mass spectrometry. Immunohistochemistry and in vivo magnetic resonance imaging were also performed. RESULTS: Sodium tungstate treatment reduced body weight gain, food intake, and blood glucose and triglyceride levels. These effects were associated with transcriptional and functional changes in the hypothalamus. Proteomic analysis revealed that sodium tungstate modified the expression levels of proteins involved in cell morphology, axonal growth, and tissue remodeling, such as actin, CRMP2 and neurofilaments, and of proteins related to energy metabolism. Moreover, immunohistochemistry studies confirmed results for some targets and further revealed tungstate-dependent regulation of SNAP25 and HPC-1 proteins, suggesting an effect on synaptogenesis as well. Functional test for cell activity based on c-fos-positive cell counting also suggested that sodium tungstate modified hypothalamic basal activity. Finally, in vivo magnetic resonance imaging showed that tungstate treatment can affect neuronal organization in the hypothalamus. CONCLUSIONS: Altogether, these results suggest that sodium tungstate regulates proteins involved in axonal and glial plasticity. The fact that sodium tungstate could modulate hypothalamic plasticity and networks in adulthood makes it a possible and interesting therapeutic strategy not only for obesity management, but also for other neurodegenerative illnesses like Alzheimer's disease

Anti-obesity sodium tungstate treatment triggers axonal and glial plasticity in hypothalamic feeding centers

Objective: This study aims at exploring the effects of sodium tungstate treatment on hypothalamic plasticity, which is known to have an important role in the control of energy metabolism. Methods: Adult lean and high-fat diet-induced obese mice were orally treated with sodium tungstate. Arcuate and paraventricular nuclei and lateral hypothalamus were separated and subjected to proteomic analysis by DIGE and mass spectrometry. Immunohistochemistry and in vivo magnetic resonance imaging were also performed. Results: Sodium tungstate treatment reduced body weight gain, food intake, and blood glucose and triglyceride levels. These effects were associated with transcriptional and functional changes in the hypothalamus. Proteomic analysis revealed that sodium tungstate modified the expression levels of proteins involved in cell morphology, axonal growth, and tissue remodeling, such as actin, CRMP2 and neurofilaments, and of proteins related to energy metabolism. Moreover, immunohistochemistry studies confirmed results for some targets and further revealed tungstate-dependent regulation of SNAP25 and HPC-1 proteins, suggesting an effect on synaptogenesis as well. Functional test for cell activity based on c-fos- positive cell counting also suggested that sodium tungstate modified hypothalamic basal activity. Finally, in vivo magnetic resonance imaging showed that tungstate treatment can affect neuronal organization in the hypothalamus. Conclusions: Altogether, these results suggest that sodium tungstate regulates proteins involved in axonal and glial plasticity. The fact that sodium tungstate could modulate hypothalamic plasticity and networks in adulthood makes it a possible and interesting therapeutic strategy not only for obesity management, but also for other neurodegenerative illnesses like Alzheimer's disease

<i>In vivo</i> sodium tungstate effects.

<p>Mice were treated with 2 g/l of sodium tungstate in water for 14 days. Body weight gain (A) and food intake (B) were measured for the duration of treatment, and blood glucose (C) and TG (D) were measured at the beginning and at the end of treatment. Open squares correspond to untreated lean mice (UL) (n = 6), black squares to treated lean mice (TL) (n = 6), open circles to untreated obese mice (UO) (n = 6), and black circles to treated obese mice (TO) (n = 6). In the bar graph, white and black bars represent untreated and treated animals, respectively. *<i>p</i><0.05; **<i>p</i><0.01; ***<i>p</i><0.001 vs. lean mice. ⁺⁺⁺<i>p</i><0.001 treated lean vs. untreated lean mice. ^#<i>p</i><0.05; ###p<0.001 treated obese vs. untreated obese mice. Data are the mean±SD.</p

Tungstate targets identified by proteomics in different hypothalamic nuclei.

a<p>Spot numbers corresponding to 2D images for each type of nuclei (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039087#pone-0039087-g002" target="_blank">Figure 2</a>).</p>b<p>Mice phenotypes: L  =  lean. O  =  obese.</p>c<p>Ratios of protein expression levels calculated using DeCyder software as the fold change in normalized spot volume comparing tungstate treated and non treated animals (Student’s <i>t</i> test based on the log of the ratio of the treated group to the control group).</p

Sodium tungstate treatment significantly modulates expression of proteins involved in neuronal plasticity.

<p>Brains from mice fed with a standard diet were sliced following 14 days of tungstate treatment and were prepared for immunoreactivity assays. (A) c-fos antibody reveals an increase in c-fos immunoreactive neurons after tungstate treatment, (B) as does the GFAP antibody. (C) The β-tubulin antibody and (D) HPC-1 show a decrease in immunoreactive neurons for these proteins. (E) NeuroD-1 immunoreactivity and (F) SNAP25 immunoreactivity increase after treatment. Scale bar  = 50 µM. White and black bars represent untreated and treated animals, respectively. **<i>p</i><0.01; ***<i>p</i><0.001. Data are the mean±SD.</p