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Benign Prostatic Hyperplasia (BPH) is the result of excessive cellullar proliferation in the prostatic transition zone, that compresses the urethra, causing the symptoms of the disease. Evidence supports that atherogenic environment could contribute to the development and progression of BPH, increasing its incidence and aggressiveness. The BPH hyperproliferative state requires coordinated communication between the different components that maintain a permissive microenvironment. Thus, extracellular vesicles (EVs) have become relevant as intercellular communication regulators in multiple processes. It has been described that oxidized-LDL molecule (OxLDL) would be involved in numerous signaling pathways; including signaling by EVs promoting cellullar proliferation. EVs, may contain diverse biomolecules that confer them functions in cellular communication. In addition, it has been reported that EVs derived from different cell types are capable to mediate proliferative and inflammatory effects. In this context, we proposed as objectives: 1) to evaluate the effect of OxLDL, simulating an atherogenic state, on cell proliferation in primary cultures of human prostate stromal cells (HPSC) from patient samples (n=8) from the Sanatorio Allende of Córdoba; 2) to isolate EVs from primary cultures by differential ultracentrifugation and analyze their production and release into the medium in treatments with OxLDL vs. vehicle; 3) to morphologically characterize EVs by transmission electron microscopy (TEM) and confirm the identity and presence of exosomes, through CD63 immuno-staining using colloidal gold. It was observed that OxLDL (20μM) produced a significant increase in cell proliferation rate compared to vehicle. EVs were obtained by differential ultracentrifugations (2k, 10k, 150k pellets) and visualized by TEM using negative staining. HPSC from patients with BPH showed a very low frequency of EVs released, with OxLDL inducing a 10-fold increase, especially in the 15-20nm fraction (p<0.001). Ultrastructurally, these EVs exhibited a spherical and concave appearance, compatible with exosomes. Therefore, they were positively verified by CD63 immunostaining. Finally, we conclude that OxLDL would favor cell proliferation, increase and release of EVs, which would participate in cell communication and maintenance of the permissive environment, propitious to progression and increased aggressiveness of BPH.La Hiperplasia Prostática Benigna (HPB) surge a consecuencia de la proliferación celular excesiva de la zona de transición prostática que comprime la uretra, provocando la sintomatología propia de la enfermedad. Evidencias sostienen que el entorno aterogénico podrían contribuir al desarrollo y progresión de la HPB, aumentando su incidencia y agresividad. El estado hiperproliferativo de la HPB requiere una coordinada comunicación entre los diferentes componentes que mantienen un microambiente permisivo. Así, las vesículas extracelulares (EVs) han tomado relevancia como reguladores de comunicación intercelular en múltiples procesos. Se ha descrito que la molécula de LDL-oxidada (OxLDL) estaría involucrada en diversas vías de señalización; incluida la señalización por EVs promoviendo la proliferación celular. Las EVs pueden contener diversas biomoléculas que les confieren funciones en comunicación intercelular y, además, se reportaron efectos proliferativos e inflamatorios mediados por EVs derivadas de distintos tipos celulares. Es este contexto, propusimos como objetivos: 1) evaluar el efecto de OxLDL, simulando un estado aterogénico, sobre la proliferación celular en cultivos primarios de células estromales prostáticas humanas (HPSC) de muestras de pacientes (n=8) del Sanatorio Allende de Córdoba; 2) aislar EVs de cultivos primarios mediante ultracentrifugaciones diferenciales y analizar su producción y liberación al medio en tratamientos con OxLDL vs. vehículo; 3) caracterizar morfológicamente EVs mediante microscopía electrónica de transmisión (TEM) y confirmar la identidad y presencia de exosomas, a través de inmuno-marcación de CD63 utilizando oro coloidal. Se observó que OxLDL (20μM) produjo un aumento significativo en la tasa de proliferación celular respecto al vehículo. EVs fueron obtenidas mediante utracentrifugaciones diferenciales (pellets 2k, 10k, 150k) y visualizadas por TEM utilizando tinción negativa. Las HPSC de pacientes con HPB mostraron una frecuencia muy baja de EVs liberadas, con OxLDL induciendo un aumento de 10 veces, especialmente en la fracción 15-20nm (p<0,001). Ultraestructuralmente, estas EVs exhibieron una apariencia esférica y cóncava, compatible con exosomas; luego fueron verificados positivamente por inmuno-marcación de CD63. Finalmente, podemos concluir que OxLDL favorecería la proliferación celular, incremento y liberación de EVs, las cuales participarían en comunicación celular y mantención del ambiente permisivo propicio para la progresión y aumento de agresividad de la HPB.

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Benign prostatic hyperplasia (BPH) is characterized by uncontrolled proliferation of mainly stromal elements, arise from a conjunction of multiple physiopathogenic factors. Among them, the metabolic syndrome components are strongly correlated to the risk of suffering from BPH and its aggressiveness, although cellular mechanisms are slightly understood. This study aimed to evaluate in vitro the proliferative effect of Oxidized-LDL (OxLDL) and the possible inhibitory role of the main therapy used for metabolic syndrome in stromal cells primary cultures derived from patients with BPH. In this sense, prostatic stromal cells from 3 patients with BPH were isolated, cultured and replicated in MCDB with 10% BFS, and thereafter frozen at -80°C for subsequent protocols. The cells were stimulated for 24 hours with OxLDL (20μM), atorvastatin (20μM and 2,5μM), metformin (10mM and 2 mM) or combination of both inhibitors, using vehicles as controls. Cell proliferation was determined by the resazurin technique and cell count. Thus, OxLDL induced a remarkable cell proliferation increase (p<0.001), with uneven effects when treated with metformin and atorvastatin. Though atorvastatin promoted a mild inhibition on OxLDL cell proliferation, it was not statistically significant. On the contrary, the metformin anti-proliferative effects were statistically significant in both doses tested (p <0.01 vs OxLDL). The combination of metformin+atorvastatin, both at low and high doses, did not improve the anti-proliferative effect of metformin alone on cell proliferation induced by OxLDL. These results confirm the pathogenic effect of OxLDL, the resulting molecule in dyslipidemic contexts, in abnormal prostate growth and suggest a direct beneficial metformin action on prostatic stromal proliferation of BPH, in the dyslipidemic environment of metabolic syndrome.La Hiperplasia Prostática Benigna (HPB) se caracteriza por la proliferación descontrolada de elementos principalmente estromales, resultado de la conjunción de múltiples factores fisiopatogénicos. Entre ellos, los componentes del síndrome metabólico se correlacionan fuertemente al riesgo de padecer HPB y a la agresividad de la misma, aunque los mecanismos celulares están poco esclarecidos. Nuestro objetivo fue evaluar in vitro el efecto proliferativo de la LDL oxidada (OxLDL) y el posible rol inhibitorio de las terapias usadas para el síndrome metabólico en cultivos primarios de células estromales provenientes de pacientes con HPB. Las células estromales prostáticas se aislaron de 3 pacientes con HPB, se cultivaron y replicaron en medio MCDB con 10%SFB y se congelaron a -80ºC para protocolos posteriores. Las células se estimularon por 24 horas con OxLDL (20μM), atorvastatina (20μM y 2,5μM), metformina (10mM y 2mM) o combinaciones de ambos inhibidores, usando vehículos como controles, determinándose la proliferación celular mediante la técnica de resazurina y por conteo celular. OxLDL indujo un marcado incremento en la proliferación celular (p<0.001), observándose efectos dispares cuando se trató con metformina y atorvastatina. Si bien atorvastatina promovió una leve disminución en la proliferación inducida por OxLDL, no resultó significativa estadísticamente. Por el contrario, la acción anti-proliferativa de metformina resultó notable, en ambas dosis ensayadas (p<0.01 vs. OxLDL). La combinación de metformina+atorvastatina, tanto a dosis bajas como altas, no mejoró la acción anti-proliferativa de metformina sola sobre la proliferación celular inducida por OxLDL. Estos resultados confirman el efecto patogénico de la OxLDL (molécula resultante de los contextos dislipidémicos) sobre el crecimiento anómalo de la próstata y sugieren una acción benéfica directa de metformina sobre la proliferación prostática estromal de la HPB en el entorno dislipidémico del síndrome metabólico.

Human Mas-related G protein-coupled receptors-X1 induce chemokine receptor 2 expression in rat dorsal root ganglia neurons and release of chemokine ligand 2 from the human LAD-2 mast cell line

Primate-specific Mas-related G protein-coupled receptors-X1 (MRGPR-X1) are highly enriched in dorsal root ganglia (DRG) neurons and induce acute pain. Herein, we analyzed effects of MRGPR-X1 on serum response factors (SRF) or nuclear factors of activated T cells (NFAT), which control expression of various markers of chronic pain. Using HEK293, DRG neuron-derived F11 cells and cultured rat DRG neurons recombinantly expressing human MRGPR-X1, we found activation of a SRF reporter gene construct and induction of the early growth response protein-1 via extracellular signal-regulated kinases-1/2 known to play a significant role in the development of inflammatory pain. Furthermore, we observed MRGPR-X1-induced up-regulation of the chemokine receptor 2 (CCR2) via NFAT, which is considered as a key event in the onset of neuropathic pain and, so far, has not yet been described for any endogenous neuropeptide. Up-regulation of CCR2 is often associated with increased release of its endogenous agonist chemokine ligand 2 (CCL2). We also found MRGPR-X1-promoted release of CCL2 in a human connective tissue mast cell line endogenously expressing MRGPR-X1. Thus, we provide first evidence to suggest that MRGPR-X1 induce expression of chronic pain markers in DRG neurons and propose a so far unidentified signaling circuit that enhances chemokine signaling by acting on two distinct yet functionally co-operating cell types. Given the important role of chemokine signaling in pain chronification, we propose that interruption of this signaling circuit might be a promising new strategy to alleviate chemokine-promoted pain

Protective role of vitamin B6 (PLP) against DNA damage in Drosophila models of type 2 diabetes

Growing evidence shows that improper intake of vitamin B6 increases cancer risk and several studies indicate that diabetic patients have a higher risk of developing tumors. We previously demonstrated that in Drosophila the deficiency of Pyridoxal 5' phosphate (PLP), the active form of vitamin B6, causes chromosome aberrations (CABs), one of cancer prerequisites, and increases hemolymph glucose content. Starting from these data we asked if it was possible to provide a link between the aforementioned studies. Thus, we tested the effect of low PLP levels on DNA integrity in diabetic cells. To this aim we generated two Drosophila models of type 2 diabetes, the first by impairing insulin signaling and the second by rearing flies in high sugar diet. We showed that glucose treatment induced CABs in diabetic individuals but not in controls. More interestingly, PLP deficiency caused high frequencies of CABs in both diabetic models demonstrating that hyperglycemia, combined to reduced PLP level, impairs DNA integrity. PLP-depleted diabetic cells accumulated Advanced Glycation End products (AGEs) that largely contribute to CABs as α-lipoic acid, an AGE inhibitor, rescued not only AGEs but also CABs. These data, extrapolated to humans, indicate that low PLP levels, impacting on DNA integrity, may be considered one of the possible links between diabetes and cancer

Externalizing behavior in early childhood and body mass index from age 2 to 12 years: longitudinal analyses of a prospective cohort study

Background: Some evidence suggests that obesity and behavior problems are related in children, but studies have been conflicting and have rarely included children under age 4. An association between behavior problems in early childhood and risk for obesity could suggest that a common set of factors contribute to both. Our research objectives were to determine the extent to which externalizing behavior in early childhood is related to body mass index (BMI) in early childhood and through age 12, and to evaluate whether these associations differ by sex and race. Methods: Data from the NICHD Study of Early Child Care and Youth Development were analyzed. Externalizing behaviors at 24 months were assessed by mothers using the Child Behavior Checklist. BMI was calculated from measured height and weight assessed 7 times between age 2 and 12 years. Linear mixed effects models were used to assess associations between 24 month externalizing behavior and BMI from 2 to 12 years, calculate predicted differences in BMI, and evaluate effect modification. Results: Externalizing behavior at 24 months was associated with a higher BMI at 24 months and through age 12. Results from a linear mixed effects model, controlling for confounding variables and internalizing behavior, predicted a difference in BMI of approximately 3/4 of a unit at 24 months of age comparing children with high levels of externalizing behavior to children with low levels of externalizing behavior. There was some evidence of effect modification by race; among white children, the average BMI difference remained stable through age 12, but it doubled to 1.5 BMI units among children who were black or another race. Conclusions: Our analyses suggest that externalizing behaviors in early childhood are associated with children's weight status early in childhood and throughout the elementary school years, though the magnitude of the effect is modest.https://doi.org/10.1186/1471-2431-10-4

Metabolic profiling of sourdough fermented wheat and rye bread

Sourdough fermentation by lactic acid bacteria is commonly used in bread baking, affecting several attributes of the final product. We analyzed whole-grain wheat and rye breads and doughs prepared with baker's yeast or a sourdough starter including Candida milleri, Lactobacillus brevis and Lactobacillus plantarum using non-targeted metabolic profiling utilizing LC-QTOF-MS. The aim was to determine the fermentation-induced changes in metabolites potentially contributing to the health-promoting properties of whole-grain wheat and rye. Overall, we identified 118 compounds with significantly increased levels in sourdough, including branched-chain amino acids (BCAAs) and their metabolites, small peptides with high proportion of BCAAs, microbial metabolites of phenolic acids and several other potentially bioactive compounds. We also identified 69 compounds with significantly decreased levels, including phenolic acid precursors, nucleosides, and nucleobases. Intensive sourdough fermentation had a higher impact on the metabolite profile of whole-grain rye compared to milder whole-grain wheat sourdough fermentation. We hypothesize that the increased amount of BCAAs and potentially bioactive small peptides may contribute to the insulin response of rye bread, and in more general, the overall protective effect against T2DM and CVD.Peer reviewe

Inadequate glucose control in type 2 diabetes is associated with impaired lung function and systemic inflammation: a cross-sectional study

<p>Abstract</p> <p>Background</p> <p>Inadequate glucose control may be simultaneously associated with inflammation and decreased lung function in type 2 diabetes. We evaluated if lung function is worse in patients with inadequate glucose control, and if inflammatory markers are simultaneously increased in these subjects.</p> <p>Methods</p> <p>Subjects were selected at the Colombian Diabetes Association Center in Bogotá. Pulmonary function tests were performed and mean residual values were obtained for forced expiratory volume (FEV₁₎, forced vital capacity (FVC) and FEV₁/FVC, with predicted values based on those derived by Hankinson et al. for Mexican-Americans. Multiple least-squares regression was used to adjust for differences in known determinants of lung function. We measured blood levels of glycosylated hemoglobin (HBA_1c), interleukin 6 (IL-6), tumor necrosis factor (TNF-α), fibrinogen, ferritin, and C-reactive protein (C-RP).</p> <p>Results</p> <p>495 diabetic patients were studied, out of which 352 had inadequate control (HBA_1c> 7%). After adjusting for known determinants of lung function, those with inadequate control had lower FEV₁(-75.4 mL, IC95%: -92, -59; P < 0.0001) and FVC (-121 mL, IC95%: -134, -108; P < 0,0001) mean residuals, and higher FEV₁/FVC (0.013%, IC95%: 0.009, 0.018, P < 0.0001) residuals than those with adequate control, as well as increased levels of all inflammatory markers (P < 0.05), with the exception of IL-6.</p> <p>Conclusions</p> <p>Subjects with type 2 diabetes and inadequate control had lower FVC and FEV₁than predicted and than those of subjects with adequate control. It is postulated that poorer pulmonary function may be associated with increased levels of inflammatory mediators.</p

Suppression of charged particle production at large transverse momentum in central Pb-Pb collisions at sNN=2.76\sqrt{s_{\rm NN}} = 2.76 TeV

Inclusive transverse momentum spectra of primary charged particles in Pb-Pb collisions at $\sqrt{s_{_{\rm NN}}}$ = 2.76 TeV have been measured by the ALICE Collaboration at the LHC. The data are presented for central and peripheral collisions, corresponding to 0-5% and 70-80% of the hadronic Pb-Pb cross section. The measured charged particle spectra in $|\eta|<0.8$ and $0.3 < p_T < 20$ GeV/$c$ are compared to the expectation in pp collisions at the same $\sqrt{s_{\rm NN}}$, scaled by the number of underlying nucleon-nucleon collisions. The comparison is expressed in terms of the nuclear modification factor $R_{\rm AA}$. The result indicates only weak medium effects ($R_{\rm AA} \approx$ 0.7) in peripheral collisions. In central collisions, $R_{\rm AA}$ reaches a minimum of about 0.14 at $p_{\rm T}=6$-7GeV/$c$ and increases significantly at larger $p_{\rm T}$. The measured suppression of high-$p_{\rm T}$ particles is stronger than that observed at lower collision energies, indicating that a very dense medium is formed in central Pb-Pb collisions at the LHC.Comment: 15 pages, 5 captioned figures, 3 tables, authors from page 10, published version, figures at http://aliceinfo.cern.ch/ArtSubmission/node/98

Two-pion Bose-Einstein correlations in central Pb-Pb collisions at sNN\sqrt{s_{\rm NN}} = 2.76 TeV

The first measurement of two-pion Bose-Einstein correlations in central Pb-Pb collisions at $\sqrt{s_{\rm NN}} = 2.76$ TeV at the Large Hadron Collider is presented. We observe a growing trend with energy now not only for the longitudinal and the outward but also for the sideward pion source radius. The pion homogeneity volume and the decoupling time are significantly larger than those measured at RHIC.Comment: 17 pages, 5 captioned figures, 1 table, authors from page 12, published version, figures at http://aliceinfo.cern.ch/ArtSubmission/node/388