RESUMEN
AIMS: Improving the composition of circulating fatty acids (FA) leads to a reduction in cardiovascular diseases (CVD) in high-risk individuals. The membrane fluidity of red blood cells (RBC), which reflects circulating FA status, may be a valid biomarker of cardiovascular (CV) risk in type 2 diabetes (T2D). METHODS: Red blood cell membrane fluidity, quantified as general polarization (GP), was assessed in 234 subjects with T2D, 86 with prior major CVD. Based on GP distribution, a cut-off of .445 was used to divide the study cohort into two groups: the first with higher GP, called GEL, and the second, defined as lower GP (LGP). Lipidomic analysis was performed to evaluate FA composition of RBC membranes. RESULTS: Although with comparable CV risk factors, the LGP group had a greater percentage of patients with major CVD than the GEL group (40% vs 24%, respectively, p < .05). Moreover, in a logistic regression analysis, a lower GP value was independently associated with the presence of macrovascular complications. Lipidomic analysis showed a clear shift of LGP membranes towards a pro-inflammatory condition due to higher content of arachidonic acid and increased omega 6/omega 3 index. CONCLUSIONS: Increased membrane fluidity is associated with a higher CV risk in subjects with T2D. If confirmed in prospective studies, membrane fluidity could be a new biomarker for residual CV risk assessment in T2D.
Asunto(s)
Enfermedades Cardiovasculares , Diabetes Mellitus Tipo 2 , Humanos , Membrana Eritrocítica/metabolismo , Fluidez de la Membrana , Estudios Prospectivos , Factores de Riesgo , Eritrocitos/metabolismo , Ácidos Grasos/metabolismo , Factores de Riesgo de Enfermedad Cardiaca , Biomarcadores/metabolismoRESUMEN
Transmembrane semaphorins are signaling molecules, controlling axonal wiring and embryo development, which are increasingly implicated in human diseases. Semaphorin 6C (Sema6C) is a poorly understood family member and its functional role is still unclear. Upon targeting Sema6C expression in a range of cancer cells, we observed dramatic growth suppression, decreased ERK phosphorylation, upregulation of cell cycle inhibitor proteins p21, p27 and p53, and the onset of cell senescence, associated with activation of autophagy. These data are consistent with a fundamental requirement for Sema6C to support viability and growth in cancer cells. Mechanistically, we unveiled a novel signaling pathway elicited by Sema6C, and dependent on its intracellular domain, mediated by tyrosine kinases c-Abl and Focal Adhesion Kinase (FAK). Sema6C was found in complex with c-Abl, and induced its phosphorylation, which in turn led to FAK activation, independent of cell-matrix adhesion. Sema6C-induced FAK activity was furthermore responsible for increased nuclear localization of YAP transcriptional regulator. Moreover, Sema6C conferred YAP signaling-dependent long-term cancer cell survival upon nutrient deprivation. In conclusion, our findings demonstrate that Sema6C elicits a cancer promoting-signaling pathway sustaining cell viability and self-renewal, independent of growth factors and nutrients availability.
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Neoplasias , Transducción de Señal , Humanos , Proteína-Tirosina Quinasas de Adhesión Focal/metabolismo , Supervivencia Celular , Quinasa 1 de Adhesión Focal/genética , Quinasa 1 de Adhesión Focal/metabolismo , Fosforilación , Proteínas de Ciclo Celular/metabolismo , Neoplasias/genéticaRESUMEN
Seminoma is the most common testicular cancer. Pituitary tumor-transforming gene 1 (PTTG1) is a securin showing oncogenic activity in several tumors. We previously demonstrated that nuclear PTTG1 promotes seminoma tumor invasion through its transcriptional activity on matrix metalloproteinase 2 (MMP-2) and E-cadherin (CDH1). We wondered if specific interactors could affect its subcellular distribution. To this aim, we investigated the PTTG1 interactome in seminoma cell lines showing different PTTG1 nuclear levels correlated with invasive properties. A proteomic approach upon PTTG1 immunoprecipitation uncovered new specific securin interactors. Western blot, confocal microscopy, cytoplasmic/nuclear fractionation, sphere-forming assay, and Atlas database interrogation were performed to validate the proteomic results and to investigate the interplay between PTTG1 and newly uncovered partners. We observed that spectrin beta-chain (SPTBN1) and PTTG1 were cofactors, with SPTBN1 anchoring the securin in the cytoplasm. SPTBN1 downregulation determined PTTG1 nuclear translocation, promoting its invasive capability. Moreover, a PTTG1 deletion mutant lacking SPTBN1 binding was strongly localized in the nucleus. The Atlas database revealed that seminomas that contained higher nuclear PTTG1 levels showed significantly lower SPTBN1 levels in comparison to non-seminomas. In human seminoma specimens, we found a strong PTTG1/SPTBN1 colocalization that decreases in areas with nuclear PTTG1 distribution. Overall, these results suggest that SPTBN1, along with PTTG1, is a potential prognostic factor useful in the clinical management of seminoma.
Asunto(s)
Seminoma , Neoplasias Testiculares , Humanos , Masculino , Línea Celular Tumoral , Citoplasma/metabolismo , Regulación Neoplásica de la Expresión Génica , Metaloproteinasa 2 de la Matriz/metabolismo , Proteómica , Securina/genética , Securina/metabolismo , Seminoma/genética , Espectrina/genética , Neoplasias Testiculares/genéticaRESUMEN
VO2max index has a significant impact on overall health. Its estimation through wearables notifies the user of his level of fitness but cannot provide a detailed analysis of the time intervals in which heartbeat dynamics are changed and/or fatigue is emerging. Here, we developed a multiple modality biosignal processing method to investigate running sessions to characterize in real time heartbeat dynamics in response to external energy demand. We isolated dynamic regimes whose fraction increases with the VO2max and with the emergence of neuromuscular fatigue. This analysis can be extremely valuable by providing personalized feedback about the user's fitness level improvement that can be realized by developing personalized exercise plans aimed to target a contextual increase in the dynamic regime fraction related to VO2max increase, at the expense of the dynamic regime fraction related to the emergence of fatigue. These strategies can ultimately result in the reduction in cardiovascular risk.
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Ejercicio Físico , Carrera , Análisis por Conglomerados , Corazón , Frecuencia Cardíaca , Consumo de Oxígeno , Aptitud Física/fisiologíaRESUMEN
In this work, we will investigate if red blood cell (RBC) membrane fluidity, influenced by several hyperglycemia-induced pathways, could provide a complementary index of HbA1c to monitor the development of type 2 diabetes mellitus (T2DM)-related macroangiopathic complications such as Peripheral Artery Disease (PAD). The contextual liquid crystalline (LC) domain spatial organization in the membrane was analysed to investigate the phase dynamics of the transition. Twenty-seven patients with long-duration T2DM were recruited and classified in DM, including 12 non-PAD patients, and DM + PAD, including 15 patients in any stage of PAD. Mean values of RBC generalized polarization (GP), representative of membrane fluidity, together with spatial organization of LC domains were compared between the two groups; p-values < 0.05 were considered statistically significant. Although comparable for anthropometric characteristics, duration of diabetes, and HbA1c, RBC membranes of PAD patients were found to be significantly more fluid (GP: 0.501 ± 0.026) than non-PAD patients (GP: 0.519 ± 0.007). These alterations were shown to be triggered by changes in both LC microdomain composition and distribution. We found a decrease in Feret diameter from 0.245 ± 0.281 µm in DM to 0.183 ± 0.124 µm in DM + PAD, and an increase in circularity. Altered RBC membrane fluidity is correlated to a spatial reconfiguration of LC domains, which, by possibly altering metabolic function, are associated with the development of T2DM-related macroangiopathic complications.
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Diabetes Mellitus Tipo 2 , Enfermedad Arterial Periférica , Diabetes Mellitus Tipo 2/complicaciones , Eritrocitos/metabolismo , Hemoglobina Glucada/metabolismo , Humanos , Fluidez de la Membrana , Enfermedad Arterial Periférica/complicacionesRESUMEN
The non-orthotopic expression of olfactory receptors (ORs) includes the male reproductive system, and in particular spermatozoa; their active ligands could be essential to sperm chemotaxis and chemical sperm-oocyte communication. OR51E2 expression has been previously reported on sperm cells' surface. It has been demonstrated in different cellular models that olfactory receptor 51E2 (OR51E2) binds volatile short-chain fatty acids (SCFAs) as specific ligands. In the present research, we make use of Western blot, confocal microscopy colocalization analysis, and the calcium-release assay to demonstrate the activation of sperm cells through OR51E2 upon SCFAs stimulus. Moreover, we perform a novel modified swim-up assay to study the involvement of OR51E2/SCFAs in sperm migration. Taking advantage of computer-assisted sperm analysis (CASA system), we determine the kinematics parameters of sperm cells migrating towards SCFAs-enriched medium, revealing that these ligands are able to promote a more linear sperm-cell orientation. Finally, we obtain SCFAs by mass spectrometry in cervico-vaginal mucus and show for the first time that a direct incubation between cervical mucus and sperm cells could promote their activation. This study can shed light on the possible function of chemosensory receptors in successful reproduction activity, laying the foundation for the development of new strategies for the treatment of infertile individuals.
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Neuronas Receptoras Olfatorias , Receptores Odorantes , Femenino , Masculino , Animales , Receptores Odorantes/metabolismo , Semen/metabolismo , Espermatozoides/metabolismo , Ácidos Grasos Volátiles , Neuronas Receptoras Olfatorias/metabolismoRESUMEN
BACKGROUND: A high level of glycosylated haemoglobin (HbA1c), which is a nonenzymatic glycosylation product, is correlated with an increased risk of developing microangiopathic complications in Diabetes Mellitus (DM). Erythrocyte membrane fluidity could provide a complementary index to monitor the development of complications since it is influenced by several hyperglycaemia-induced pathways and other independent risk factors. MATERIALS AND METHODS: 15 healthy controls and 33 patients with long-duration (≥20 years) type 1 Diabetes Mellitus (T1DM) were recruited. Diabetic subjects were classified into two groups: T1DM, constituted by 14 nonretinopathic patients, and T1DM + RD, constituted by 19 patients in any stage of diabetic retinopathy. Red blood cells (RBC) were incubated with the fluorescent Laurdan probe and median values of Generalized Polarization (GP), representative of membrane fluidity, were compared between the two groups. Baseline characteristics among groups have been compared with Student's t test or ANOVA. Values of P < .05 were considered statistically significant. RESULTS: All the participants were comparable for age, Body Mass Index (BMI), creatinine and lipid profile. The duration of diabetes was similar for T1DM (34.4 ± 7.8 years) and T1DM + RD (32.8 ± 7.5 years) subjects as well as values of HbA1c: (55.6 ± 8.1) mmol/mol for T1DM and (61.2 ± 11.0) mmol/mol for T1DM + RD, respectively. Erythrocyte plasmatic membranes of RD patients were found to be more fluid (GP: 0.40 ± 0.04) than non-RD patients (GP: 0.43 ± 0.03) with a statistically significant difference (P = .035). CONCLUSIONS: Altered erythrocyte membrane fluidity may therefore represent a marker of retinopathy in T1DM patients as a result of post-translational modifications of multifactorial aetiology (nonenzymatic glycosylation of proteins, generation of reactive oxygen species, lipid peroxidation).
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Diabetes Mellitus Tipo 1/sangre , Retinopatía Diabética/sangre , Membrana Eritrocítica/metabolismo , Eritrocitos/metabolismo , Fluidez de la Membrana/fisiología , Adulto , Biomarcadores , Estudios de Casos y Controles , Diabetes Mellitus Tipo 1/complicaciones , Retinopatía Diabética/etiología , Membrana Eritrocítica/fisiología , Femenino , Hemoglobina Glucada/metabolismo , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Free fatty acids are essential structural components of the cell, and their intracellular distribution and effects on membrane organelles have crucial roles in regulating the metabolism, development, and cell cycle of most cell types. Here we engineered novel fluorescent, polarity-sensitive fatty acid derivatives, with the fatty acid aliphatic chain of increasing length (from 12 to 18 carbons). As in the laurdan probe, the lipophilic acyl tail is connected to the environmentally sensitive dimethylaminonaphthalene moiety. The fluorescence lifetime imaging analysis allowed us to monitor the intracellular distribution of the free fatty acids within the cell, and to simultaneously examine how the fluidity and the microviscosity of the membrane environment influence their localization. Each of these probes can thus be used to investigate the membrane fluidity regulation of the correspondent fatty acid intracellular distribution. We observed that, in PC-12 cells, fluorescent sensitive fatty acid derivatives with increased chain length compartmentalize more preferentially in the fluid regions, characterized by a low microviscosity. Moreover, fatty acid derivatives with the longest chain compartmentalize in lipid droplets and lysosomes with characteristic lifetimes, thus making these probes a promising tool for monitoring lipophagy and related events.
Asunto(s)
Ácidos Grasos/metabolismo , Colorantes Fluorescentes/metabolismo , Espacio Intracelular/metabolismo , Fluidez de la Membrana , 2-Naftilamina/análogos & derivados , 2-Naftilamina/química , Animales , Fluorescencia , Lauratos/química , Lisosomas/metabolismo , Células PC12 , Ratas , Solventes , ViscosidadRESUMEN
BACKGROUND: Deregulation of the tricarboxylic acid cycle (TCA) due to mutations in specific enzymes or defective aerobic metabolism is associated with tumour growth. Aconitase 2 (ACO2) participates in the TCA cycle by converting citrate to isocitrate, but no evident demonstrations of its involvement in cancer metabolism have been provided so far. METHODS: Biochemical assays coupled with molecular biology, in silico, and cellular tools were applied to circumstantiate the impact of ACO2 in the breast cancer cell line MCF-7 metabolism. Fluorescence lifetime imaging microscopy (FLIM) of NADH was used to corroborate the changes in bioenergetics. RESULTS: We showed that ACO2 levels are decreased in breast cancer cell lines and human tumour biopsies. We generated ACO2- overexpressing MCF-7 cells and employed comparative analyses to identify metabolic adaptations. We found that increased ACO2 expression impairs cell proliferation and commits cells to redirect pyruvate to mitochondria, which weakens Warburg-like bioenergetic features. We also demonstrated that the enhancement of oxidative metabolism was supported by mitochondrial biogenesis and FoxO1-mediated autophagy/mitophagy that sustains the increased ROS burst. CONCLUSIONS: This work identifies ACO2 as a relevant gene in cancer metabolic rewiring of MCF-7 cells, promoting a different utilisation of pyruvate and revealing the potential metabolic vulnerability of ACO2-associated malignancies.
Asunto(s)
Aconitato Hidratasa/genética , Neoplasias de la Mama/genética , Proteína Forkhead Box O1/genética , Estrés Oxidativo/efectos de los fármacos , Aconitato Hidratasa/antagonistas & inhibidores , Autofagia/efectos de los fármacos , Autofagia/genética , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/patología , Proliferación Celular/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Femenino , Humanos , Células MCF-7 , Mitocondrias/efectos de los fármacos , Mitocondrias/genética , Mutación , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Fragile X-related disorders are due to a dynamic mutation of the CGG repeat at the 5' UTR of the FMR1 gene, coding for the RNA-binding protein FMRP. As the CGG sequence expands from premutation (PM, 56-200 CGGs) to full mutation (> 200 CGGs), FMRP synthesis decreases until it is practically abolished in fragile X syndrome (FXS) patients, mainly due to FMR1 methylation. Cells from rare individuals with no intellectual disability and carriers of an unmethylated full mutation (UFM) produce slightly elevated levels of FMR1-mRNA and relatively low levels of FMRP, like in PM carriers. With the aim of clarifying how UFM cells differ from CTRL and FXS cells, a comparative proteomic approach was undertaken, from which emerged an overexpression of SOD2 in UFM cells, also confirmed in PM but not in FXS. The SOD2-mRNA bound to FMRP in UFM more than in the other cell types. The high SOD2 levels in UFM and PM cells correlated with lower levels of superoxide and reactive oxygen species (ROS), and with morphological anomalies and depolarization of the mitochondrial membrane detected through confocal microscopy. The same effect was observed in CTRL and FXS after treatment with MC2791, causing SOD2 overexpression. These mitochondrial phenotypes reverted after knock-down with siRNA against SOD2-mRNA and FMR1-mRNA in UFM and PM. Overall, these data suggest that in PM and UFM carriers, which have high levels of FMR1 transcription and may develop FXTAS, SOD2 overexpression helps to maintain low levels of both superoxide and ROS with signs of mitochondrial degradation.
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Ataxia/patología , Metilación de ADN , Proteína de la Discapacidad Intelectual del Síndrome del Cromosoma X Frágil/metabolismo , Síndrome del Cromosoma X Frágil/patología , Mitocondrias/patología , Proteínas Mitocondriales/metabolismo , Mutación , Proteoma/análisis , Temblor/patología , Ataxia/genética , Ataxia/metabolismo , Estudios de Casos y Controles , Células Cultivadas , Fibroblastos/metabolismo , Fibroblastos/patología , Proteína de la Discapacidad Intelectual del Síndrome del Cromosoma X Frágil/genética , Síndrome del Cromosoma X Frágil/genética , Síndrome del Cromosoma X Frágil/metabolismo , Humanos , Masculino , Mitocondrias/metabolismo , Proteínas Mitocondriales/genética , ARN Interferente Pequeño/genética , Superóxido Dismutasa/antagonistas & inhibidores , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismo , Temblor/genética , Temblor/metabolismoRESUMEN
Palmitic acid metabolism involves delta-9 and delta-6 desaturase enzymes forming palmitoleic acid (9cis-16:1; n-7 series) and sapienic acid (6cis-16:1; n-10 series), respectively. The corresponding biological consequences and lipidomic research on these positional monounsaturated fatty acid (MUFA) isomers are under development. Furthermore, sapienic acid can bring to the de novo synthesis of the n-10 polyunsaturated fatty acid (PUFA) sebaleic acid (5cis,8cis-18:2), but such transformations in cancer cells are not known. The model of Caco-2 cell line was used to monitor sapienic acid supplementation (150 and 300 µM) and provide evidence of the formation of n-10 fatty acids as well as their incorporation at levels of membrane phospholipids and triglycerides. Comparison with palmitoleic and palmitic acids evidenced that lipid remodelling was influenced by the type of fatty acid and positional isomer, with an increase of 8cis-18:1, n-10 PUFA and a decrease of saturated fats in case of sapienic acid. Cholesteryl esters were formed only in cases with sapienic acid. Sapienic acid was the less toxic among the tested fatty acids, showing the highest EC50s and inducing death only in 75% of cells at the highest concentration tested. Two-photon fluorescent microscopy with Laurdan as a fluorescent dye provided information on membrane fluidity, highlighting that sapienic acid increases the distribution of fluid regions, probably connected with the formation of 8cis-18:1 and the n-10 PUFA in cell lipidome. Our results bring evidence for MUFA positional isomers and de novo PUFA synthesis for developing lipidomic analysis and cancer research.
Asunto(s)
Neoplasias del Colon/metabolismo , Ácidos Grasos Omega-3/metabolismo , Ácidos Palmíticos/metabolismo , Fosfolípidos/química , Células CACO-2 , Membrana Celular/química , Membrana Celular/metabolismo , Ésteres del Colesterol/biosíntesis , Ésteres del Colesterol/química , Ésteres del Colesterol/metabolismo , Neoplasias del Colon/química , Neoplasias del Colon/patología , Ácidos Grasos Monoinsaturados/química , Ácidos Grasos Monoinsaturados/farmacología , Ácidos Grasos Omega-3/biosíntesis , Humanos , Ácidos Linoleicos/química , Ácidos Linoleicos/metabolismo , Ácidos Linoleicos/farmacología , Linoleoil-CoA Desaturasa/química , Microscopía Fluorescente , Ácido Palmítico/química , Ácido Palmítico/metabolismo , Ácidos Palmíticos/química , Ácidos Palmíticos/farmacología , Fosfolípidos/biosíntesisRESUMEN
Modern omics techniques reveal molecular structures and cellular networks of tissues and cells in unprecedented detail. Recent advances in single cell analysis have further revolutionized all disciplines in cellular and molecular biology. These methods have also been employed in current investigations on the structure and function of insulin secreting beta cells under normal and pathological conditions that lead to an impaired glucose tolerance and type 2 diabetes. Proteomic and transcriptomic analyses have pointed to significant alterations in protein expression and function in beta cells exposed to diabetes like conditions (e.g., high glucose and/or saturated fatty acids levels). These nutritional overload stressful conditions are often defined as glucolipotoxic due to the progressive damage they cause to the cells. Our recent studies on the rat insulinoma-derived INS-1E beta cell line point to differential effects of such conditions in the phospholipid bilayers in beta cells. This review focuses on confocal microscopy-based detection of these profound alterations in the plasma membrane and membranes of insulin granules and lipid droplets in single beta cells under such nutritional load conditions.
Asunto(s)
Diabetes Mellitus Tipo 2/metabolismo , Ácidos Grasos/metabolismo , Intolerancia a la Glucosa/metabolismo , Glucosa/metabolismo , Células Secretoras de Insulina/metabolismo , Insulina/metabolismo , Animales , Línea Celular Tumoral , Membrana Celular/química , Membrana Celular/metabolismo , Gránulos Citoplasmáticos/metabolismo , Gránulos Citoplasmáticos/patología , Diabetes Mellitus Tipo 2/fisiopatología , Glucosa/farmacología , Intolerancia a la Glucosa/fisiopatología , Humanos , Células Secretoras de Insulina/química , Células Secretoras de Insulina/patología , Gotas Lipídicas/metabolismo , Gotas Lipídicas/patología , Metabolismo de los Lípidos , Lipidómica/métodos , Fosfolípidos/metabolismo , Ratas , Análisis de la Célula IndividualRESUMEN
Organisms store fatty acids in triacylglycerols in the form of lipid droplets, or hydrolyze triacylglycerols in response to energetic demands via activation of lipolytic or storage pathways. These pathways are complex sets of sequential reactions that are finely regulated in different cell types. Here we present a high spatial and temporal resolution-based method for the quantification of the turnover of fatty acids into triglycerides in live cells without introducing sample preparation artifacts. We performed confocal spectral imaging of intracellular micropolarity in cultured insulin secreting beta cells to detect micropolarity variations as they occur in time and at different pixels of microscope images. Acquired data are then analyzed in the framework of the spectral phasors technique. The method furnishes a metabolic parameter, which quantitatively assesses fatty acids - triacylglycerols turnover and the activation of lipolysis and storage pathways. Moreover, it provides a polarity profile, which represents the contribution of hyperpolar, polar and non-polar classes of lipids. These three different classes can be visualized on the image at a submicrometer resolution, revealing the spatial localization of lipids in cells under physiological and pathological settings. This new method allows for a fine-tuned, real-time visualization of the turnover of fatty acids into triglycerides in live cells with submicrometric resolution. It also detects imbalances between lipid storage and usage, which may lead to metabolic disorders within living cells and organisms.
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Polaridad Celular , Microscopía Intravital/métodos , Lípidos/análisis , Lipólisis , Células 3T3-L1 , Animales , Microscopía Intravital/instrumentación , Ratones , Microscopía Confocal/instrumentación , Microscopía Confocal/métodos , Análisis Espectral/instrumentación , Análisis Espectral/métodosRESUMEN
BACKGROUND: Noise exposure impairs outer hair cells (OHCs). The common basis for OHC dysfunction and loss by acoustic over-stimulation is represented by reactive oxygen species (ROS) overload that may affect the membrane structural organization through generation of lipid peroxidation. METHODS: Here we investigated in OHC different functional zones the mechanisms linking metabolic functional state (NAD(P)H intracellular distribution) to the generation of lipid peroxides and to the physical state of membranes by two photon fluorescence microscopy. RESULTS: In OHCs of control animals, a more oxidized NAD(P)H redox state is associated to a less fluid plasma membrane structure. Acoustic trauma induces a topologically differentiated NAD(P)H oxidation in OHC rows, which is damped between 1 and 6h. Peroxidation occurs after ~4h from noise insult, while ROS are produced in the first 0.2h and damage cells for a period of time after noise exposure has ended (~7.5h) when a decrease of fluidity of OHC plasma membrane occurs. OHCs belonging to inner rows, characterized by a lower metabolic activity with respect to other rows, show less severe metabolic impairment. CONCLUSIONS: Our data indicate that plasma membrane fluidity is related to NAD(P)H redox state and lipid peroxidation in hair cells. GENERAL SIGNIFICANCE: Our results could pave the way for therapeutic intervention targeting the onset of redox umbalance.
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Membrana Celular/metabolismo , Células Ciliadas Auditivas Externas/metabolismo , Fluidez de la Membrana , Ruido/efectos adversos , Animales , Oído Externo/metabolismo , Oído Externo/patología , Células Ciliadas Auditivas Externas/patología , Pérdida Auditiva Provocada por Ruido/metabolismo , Pérdida Auditiva Provocada por Ruido/fisiopatología , Peroxidación de Lípido , NADP/metabolismo , Especies Reactivas de Oxígeno/metabolismoRESUMEN
We have generated unique asymmetric liposomes with phosphatidylserine (PS) distributed at the outer membrane surface to resemble apoptotic bodies and phosphatidic acid (PA) at the inner layer as a strategy to enhance innate antimycobacterial activity in phagocytes while limiting the inflammatory response. Results show that these apoptotic body-like liposomes carrying PA (ABL/PA) (i) are more efficiently internalized by human macrophages than by nonprofessional phagocytes, (ii) induce cytosolic Ca(2+) influx, (iii) promote Ca(2+)-dependent maturation of phagolysosomes containing Mycobacterium tuberculosis (MTB), (iv) induce Ca(2+)-dependent reactive oxygen species (ROS) production, (v) inhibit intracellular mycobacterial growth in differentiated THP-1 cells as well as in type-1 and -2 human macrophages, and (vi) down-regulate tumor necrosis factor (TNF)-α, interleukin (IL)-12, IL-1ß, IL-18, and IL-23 and up-regulate transforming growth factor (TGF)-ß without altering IL-10, IL-27, and IL-6 mRNA expression. Also, ABL/PA promoted intracellular killing of M. tuberculosis in bronchoalveolar lavage cells from patients with active pulmonary tuberculosis. Furthermore, the treatment of MTB-infected mice with ABL/PA, in combination or not with isoniazid (INH), dramatically reduced lung and, to a lesser extent, liver and spleen mycobacterial loads, with a concomitant 10-fold reduction of serum TNF-α, IL-1ß, and IFN-γ compared with that in untreated mice. Altogether, these results suggest that apoptotic body-like liposomes may be used as a Janus-faced immunotherapeutic platform to deliver polar secondary lipid messengers, such as PA, into phagocytes to improve and recover phagolysosome biogenesis and pathogen killing while limiting the inflammatory response.
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Liposomas/farmacología , Macrófagos/inmunología , Macrófagos/microbiología , Mycobacterium tuberculosis/inmunología , Tuberculosis Pulmonar/tratamiento farmacológico , Tuberculosis Pulmonar/inmunología , Adulto , Animales , Antituberculosos/farmacología , Apoptosis/inmunología , Líquido del Lavado Bronquioalveolar/citología , Líquido del Lavado Bronquioalveolar/inmunología , Calcio/metabolismo , Línea Celular Tumoral , Modelos Animales de Enfermedad , Humanos , Inmunidad Innata/inmunología , Isoniazida/farmacología , Leucemia Monocítica Aguda , Liposomas/inmunología , Liposomas/metabolismo , Macrófagos/citología , Masculino , Ratones , Ratones Endogámicos BALB C , Persona de Mediana Edad , Fagocitosis/inmunología , Fosfatidilserinas/metabolismo , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Cell motility involves a number of strategies that cells use in order to seek nutrients, escape danger, and fulfill morphogenetic roles. Here we present a methodology to quantify morphological changes and their relationship with signaling events from time-lapse imaging microscopy experiments, in order to characterize physiological and pathological processes. To this aim, the stationary spatial pattern of signaling events is determined through an intracellular fluorescent probe, and it is related with the frequency and entity of morphodynamic events, which are in turn quantified through a stochastic approach: two pseudoimages are obtained from a time series of moving cells that describe the probability that a pixel belongs to the cell, and the probability that a pixel is subject to a dynamic event. The simultaneous construction of these maps permits visualization of hot spots of dynamic events, i.e., zones of formation of membrane protrusions and retractions and their relationship with the signaling events reported by the specific probe employed. The method is tested on spontaneous movement of cells, trasfected with redox-sensitive yellow fluorescent protein, in which the distribution of the hot spots and its change upon expression of constitutively active Rac (V12-Rac), is related to the distribution of oxidized spots.
Asunto(s)
Ensayos de Migración Celular/métodos , Movimiento Celular , Microscopía Fluorescente/métodos , Transducción de Señal , Animales , Línea Celular Tumoral , Ratones , Coloración y Etiquetado , Imagen de Lapso de TiempoRESUMEN
The rising obesity epidemic requires effective and sustainable weight loss intervention strategies that take into account both of individual preferences and environmental impact. This study aims to develop and evaluate the effectiveness of an innovative digital biohacking approach for dietary modifications in promoting sustainable weight loss and reducing carbon footprint impact. A pilot study was conducted involving four participants who monitored their weight, diet, and activities over the course of a year. Data on food consumption, carbon footprint impact, calorie intake, macronutrient composition, weight, and energy expenditure were collected. A digital replica of the metabolism based on nutritional information, the Personalized Metabolic Avatar (PMA), was used to simulate weight changes, plan, and execute the digital biohacking approach to dietary interventions. The dietary modifications suggested by the digital biohacking approach resulted in an average daily calorie reduction of 236.78 kcal (14.24%) and a 15.12% reduction in carbon footprint impact (-736.48 gCO2eq) per participant. Digital biohacking simulations using PMA showed significant differences in weight change compared to actual recorded data, indicating effective weight reduction with the digital biohacking diet. Additionally, linear regression analysis on real data revealed a significant correlation between adherence to the suggested diet and weight loss. In conclusion, the digital biohacking recommendations provide a personalized and sustainable approach to weight loss, simultaneously reducing calorie intake and minimizing the carbon footprint impact. This approach shows promise in combating obesity while considering both individual preferences and environmental sustainability.
Asunto(s)
Huella de Carbono , Ingestión de Energía , Obesidad , Pérdida de Peso , Humanos , Proyectos Piloto , Masculino , Femenino , Obesidad/dietoterapia , Adulto , Metabolismo Energético , Persona de Mediana Edad , Dieta Reductora/métodos , Dieta/métodosRESUMEN
The objective of this review was to critically examine existing digital applications, tailored for use by citizens and professionals, to provide diet monitoring, diet planning, and precision nutrition. We sought to identify the strengths and weaknesses of such digital applications, while exploring their potential contributions to enhancing public health, and discussed potential developmental pathways. Nutrition is a critical aspect of maintaining good health, with an unhealthy diet being one of the primary risk factors for chronic diseases, such as obesity, diabetes, and cardiovascular disease. Tracking and monitoring one's diet has been shown to help improve health and weight management. However, this task can be complex and time-consuming, often leading to frustration and a lack of adherence to dietary recommendations. Digital applications for diet monitoring, diet generation, and precision nutrition offer the promise of better health outcomes. Data on current nutrition-based digital tools was collected from pertinent literature and software providers. These digital tools have been designed for particular user groups: citizens, nutritionists, and physicians and researchers employing genetics and epigenetics tools. The applications were evaluated in terms of their key functionalities, strengths, and limitations. The analysis primarily concentrated on artificial intelligence algorithms and devices intended to streamline the collection and organization of nutrition data. Furthermore, an exploration was conducted of potential future advancements in this field. Digital applications designed for the use of citizens allow diet self-monitoring, and they can be an effective tool for weight and diabetes management, while digital precision nutrition solutions for professionals can provide scalability, personalized recommendations for patients, and a means of providing ongoing diet support. The limitations in using these digital applications include data accuracy, accessibility, and affordability, and further research and development are required. The integration of artificial intelligence, machine learning, and blockchain technology holds promise for improving the performance, security, and privacy of digital precision nutrition interventions. Multidisciplinarity is crucial for evidence-based and accessible solutions. Digital applications for diet monitoring and precision nutrition have the potential to revolutionize nutrition and health. These tools can make it easier for individuals to control their diets, help nutritionists provide better care, and enable physicians to offer personalized treatment.
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Polyphenols are a class of natural compounds that act as antioxidants, neutralising harmful free radicals that would damage cells and increase the risk of diseases such as cancer, diabetes and heart disease. They also reduce inflammation, which is thought to be at the root of many chronic diseases. We are investigating the photoprotective effects of punicalagin, a type of polyphenolic compound mainly found in pomegranates, against UVA-induced damage in human skin fibroblasts. Punicalagin increases cell viability and reduces the high levels of ROS generated by photooxidative stress through its ability to modulate the Nrf2 transcriptional pathway. Interestingly, activation of the Nrf2 pathway results in an increase in reduced glutathione, NADH, and subsequently protects mitochondrial respiratory capacity. Integrating molecular and imaging approaches, our results demonstrate a potential cytoprotective effect of punicalagin against UVA-induced skin damage through an anti-apoptotic mechanism.
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OBJECTIVE: Maintaining plasma glucose homeostasis is vital for mammalian survival, but the masticatory function, which influences glucose regulation, has, to our knowledge, been overlooked. RESEARCH METHODS AND PROCEDURES: In this study, we investigated the relationship between the glycemic response curve and chewing performance in a group of 8 individuals who consumed 80 g of apple. A device called "Chewing" utilizing electromyographic (EMG) technology quantitatively assesses chewing pattern, while glycemic response is analyzed using continuous glucose monitoring. We assessed chewing pattern characterizing chewing time (tchew), number of bites (nchew), work (w), power (wr), and chewing cycles (tcyc). Moreover, we measured the principal features of the glycemic response curve, including the area under the curve (α) and the mean time to reach the glycemic peak (tmean). We used linear regression models to examine the correlations between these variables. RESULTS: tchew, nchew, and wr were correlated with α (R2 = â 0.44, â â Pâ < â 0.05â for tchew and nchew, Pâ < â 0.001 for wr), and tmean was correlated withâ tchew (R2â = â 0.25, â Pâ < â 0.05). These findings suggest that increasing chewing time and power, while reducing the number of chews, resulted in a wider glycemic curve and an earlier attainment of the glycemic peak. CONCLUSIONS: These results emphasize the influence of proper chewing techniques on blood sugar levels. Implementing correct chewing habits could serve as an additional approach to managing the glycemic curve, particularly for individuals with diabetes.