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OBJECTIVES: A timely diagnosis is imperative for curing cancer. However, in patients with rheumatic musculoskeletal diseases (RMDs) or paraneoplastic syndromes, misleading symptoms frequently delay cancer diagnosis. As metabolic remodelling characterises both cancer and RMD, we analysed if a metabolic signature can indicate paraneoplasia (PN) or reveal concomitant cancer in patients with RMD. METHODS: Metabolic alterations in the sera of rheumatoid arthritis (RA) patients with (n=56) or without (n=52) a history of invasive cancer were quantified by nuclear magnetic resonance analysis. Metabolites indicative of cancer were determined by multivariable regression analyses. Two independent RA and spondyloarthritis (SpA) cohorts with or without a history of invasive cancer were used for blinded validation. Samples from patients with active cancer or cancer treatment, pulmonary and lymphoid type cancers, paraneoplastic syndromes, non-invasive (NI) precancerous lesions and non-melanoma skin cancer and systemic lupus erythematosus and samples prior to the development of malignancy were used to test the model performance. RESULTS: Based on the concentrations of acetate, creatine, glycine, formate and the lipid ratio L1/L6, a diagnostic model yielded a high sensitivity and specificity for cancer diagnosis with AUC=0.995 in the model cohort, AUC=0.940 in the blinded RA validation cohort and AUC=0.928 in the mixed RA/SpA cohort. It was equally capable of identifying cancer in patients with PN. The model was insensitive to common demographic or clinical confounders or the presence of NI malignancy like non-melanoma skin cancer. CONCLUSIONS: This new set of metabolic markers reliably predicts the presence of cancer in arthritis or PN patients with high sensitivity and specificity and has the potential to facilitate a rapid and correct diagnosis of malignancy.
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Artritis Reumatoide , Metaboloma , Neoplasias , Síndromes Paraneoplásicos , Humanos , Masculino , Femenino , Persona de Mediana Edad , Artritis Reumatoide/sangre , Artritis Reumatoide/complicaciones , Artritis Reumatoide/diagnóstico , Neoplasias/sangre , Neoplasias/complicaciones , Síndromes Paraneoplásicos/sangre , Síndromes Paraneoplásicos/diagnóstico , Anciano , Adulto , Enfermedades Reumáticas/sangre , Enfermedades Reumáticas/complicaciones , Sensibilidad y Especificidad , Biomarcadores de Tumor/sangreRESUMEN
BACKGROUND: Infertility is a major health issue, affecting 15% of reproductive-age couples with male factors contributing to 50% of cases. Asthenozoospermia (AS), or low sperm motility, is a common cause of male infertility with complex aetiology, involving genetic and metabolic alterations, inflammation and oxidative stress. However, the molecular mechanisms behind low motility are unclear. In this study, we used a metabolomics approach to identify metabolic biomarkers and pathways involved in sperm motility. METHODS: We compared the metabolome and lipidome of spermatozoa of men with normozoospermia (n = 44) and AS (n = 22) using untargeted LC-MS and the metabolome of seminal fluid using 1H-NMR. Additionally, we evaluated the seminal fluid redox status to assess the oxidative stress in the ejaculate. RESULTS: We identified 112 metabolites and 209 lipids in spermatozoa and 27 metabolites in the seminal fluid of normozoospermic and asthenozoospermic men. PCA analysis of the spermatozoa's metabolomics and lipidomics data showed a clear separation between groups. Spermatozoa of asthenozoospermic men presented lower levels of several amino acids, and increased levels of energetic substrates and lysophospholipids. However, the metabolome and redox status of the seminal fluid was not altered inAS. CONCLUSIONS: Our results indicate impaired metabolic pathways associated with redox homeostasis and amino acid, energy and lipid metabolism in AS. Taken together, these findings suggest that the metabolome and lipidome of human spermatozoa are key factors influencing their motility and that oxidative stress exposure during spermatogenesis or sperm maturation may be in the aetiology of decreased motility in AS.
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BACKGROUND: Pedal arch interrogation by ultrasound through systolic acceleration time measurement (Pedal Acceleration Time [PAT]) is a recently described technique that offers valuable insights into foot hemodynamics. Previous studies have demonstrated a correlation between PAT and the ankle-brachial index and PAT has been proposed as a prognostic tool for wound healing in patients with peripheral artery disease. This study aims to assess the correlation between PAT and Transcutaneous Pressure of Oxygen (TcPO2) in the diabetic foot population. METHODS: In this single-center cross-sectional study PAT and TcPO2 were measured in diabetic patients with ischemic foot ulcers. The assessment of wound status was performed between the 12th and 16th week after the first evaluation. The primary outcome was to assess the correlation between the values obtained for PAT and TcPO2. The secondary outcome was to establish a cut-off value for wound healing. RESULTS: Sixty limbs with ischemic foot ulcers were evaluated. The results showed a correlation between PAT and TcPO2 [F (1.52) = 16.928; R2 = 0.246; P < 0.001]. The receiver operating characteristics curve analysis showed an optimal cut-off value at PAT >186 ms for predicting a nonhealing ulcer with a 96% specificity (area under the curve = 0.774; confidence interval 0.648-0.872; P < 0.001). CONCLUSIONS: PAT showed a significant correlation with TcPO2 with high specificity to identify nonhealing foot ulcers due to insufficient foot perfusion with a potential prognostic value in the diabetic foot population.
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Monitoreo de Gas Sanguíneo Transcutáneo , Pie Diabético , Valor Predictivo de las Pruebas , Cicatrización de Heridas , Humanos , Pie Diabético/fisiopatología , Pie Diabético/diagnóstico , Pie Diabético/terapia , Masculino , Femenino , Estudios Transversales , Anciano , Persona de Mediana Edad , Factores de Tiempo , Pie/irrigación sanguínea , Oxígeno/sangre , Flujo Sanguíneo Regional , Isquemia/fisiopatología , Isquemia/diagnóstico por imagen , Isquemia/diagnóstico , Isquemia/terapia , HemodinámicaRESUMEN
Foot ulceration and infection is associated with a substantial increase in morbidity and mortality in patients with diabetes. We present a clinical case of recurrent diabetic foot infection with an atypical clinical evolution. A 58-year-old male patient with type 1 diabetes and a history of bilateral Charcot foot neuroarthropathy was followed at our Diabetic Foot Clinic for an unhealed plantar foot ulcer for >1.5 years with recurrent episodes of infection. He was admitted to hospital due to foot ulcer reinfection with sepsis and ipsilateral lower limb cellulitis. The foot infection was found to be associated with an underlying abscess in the anterior compartment of the leg, with a cutaneous fistulous course with extensive alterations of an inflammatory nature. Exudate from the lesion was drained and tissue biopsied, revealing Serratia marcescens and Klebsiella oxytoca with dystrophic calcification (DC). Surgical excision of dystrophic tissue with debridement of the fistulous tracts was performed. The excised material corroborated the presence of fibroadipose connective tissue with marked DC, as well as areas of mixed inflammation compatible with a chronic infectious aetiology. Targeted long-term antibiotic therapy was implemented, for a total of six weeks, with a favourable clinical evolution and complete closure of the lesion at the final follow-up. DC results from calcium deposition in degenerated tissues without evidence of systemic mineral imbalance and is a potential cause of non-healing ulcers. Few cases of DC have been reported in diabetic foot patients and its treatment remains challenging and controversial. A longer follow-up period is necessary to verify the effectiveness of our approach.
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Calcinosis , Diabetes Mellitus , Pie Diabético , Sepsis , Enfermedades de la Piel , Masculino , Humanos , Persona de Mediana Edad , Pie Diabético/complicaciones , Pierna , Absceso , Calcinosis/complicacionesRESUMEN
In arterial hypertension, the dysregulation of several metabolic pathways is closely associated with chronic immune imbalance and inflammation progression. With time, these disturbances lead to the development of progressive disease and end-organ involvement. However, the influence of cholecalciferol on metabolic pathways as a possible mechanism of its immunomodulatory activity in obesity-related hypertension is not known. In a phase 2, randomized, single-center, 24-week trial, we evaluated, as a secondary outcome, the serum metabolome of 36 age- and gender-matched adults with obesity-related hypertension and vitamin D deficiency, before and after supplementation with cholecalciferol therapy along with routine medication. The defined endpoint was the assessment of circulating metabolites using a nuclear magnetic resonance-based metabolomics approach. Univariate and multivariate analyses were used to evaluate the systemic metabolic alterations caused by cholecalciferol. In comparison with normotensive controls, hypertensive patients presented overall decreased expression of several amino acids (p < 0.05), including amino acids with ketogenic and glucogenic properties as well as aromatic amino acids. Following cholecalciferol supplementation, increases were observed in glutamine (p < 0.001) and histidine levels (p < 0.05), with several other amino acids remaining unaffected. Glucose (p < 0.05) and acetate (p < 0.05) decreased after 24 weeks in the group taking the supplement, and changes in the saturation of fatty acids (p < 0.05) were also observed, suggesting a role of liposoluble vitamin D in lipid metabolism. Long-term cholecalciferol supplementation in chronically obese and overweight hypertensives induced changes in the blood serum metabolome, which reflected systemic metabolism and may have fostered a new microenvironment for cell proliferation and biology. Of note, the increased availability of glutamine may be relevant for the proliferation of different T-cell subsets.
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Hipertensión , Deficiencia de Vitamina D , Adulto , Humanos , Colecalciferol/farmacología , Colecalciferol/uso terapéutico , Glutamina/uso terapéutico , Glucosa/uso terapéutico , Vitamina D/uso terapéutico , Obesidad/complicaciones , Obesidad/tratamiento farmacológico , Suplementos Dietéticos , Deficiencia de Vitamina D/complicaciones , Hipertensión/complicaciones , Hipertensión/tratamiento farmacológico , Aminoácidos/metabolismo , Método Doble CiegoRESUMEN
OBJECTIVES: Spirituality, emotional intelligence, and palliative care (PC) knowledge have a positive and direct influence on self-efficacy and on perception of preparation and ability to provide end-of-life (EOL) care. The aim of this work is to propose a conceptual model that relates spirituality, emotional intelligence, PC knowledge, self-efficacy, and the preparation and ability to provide EOL care by doctors and nurses. METHODS: Quantitative, exploratory, descriptive, and inferential study applied to doctors and nurses in a hospital in the north of Portugal, between May and July 2022. The data collection instrument includes a questionnaire. The relationships between latent variables were evaluated using structural equation models by the partial least squares method using the Smart PLS 3.0 software. It was obtained the previous authorization of the ethics committee. RESULTS: The results (n = 380) indicate that self-efficacy, spirituality, and PC knowledge have a positive influence on the ability to provide EOL care. Emotional intelligence and spirituality have a direct and positive effect on self-efficacy. There is no direct influence of emotional intelligence on the ability to provide EOL care, but emotional intelligence has an indirect effect mediated by self-efficacy. SIGNIFICANCE OF RESULTS: Spirituality, self-efficacy, and emotional intelligence are very important for the ability of doctors and nurses to provide EOL care. The identification of predictive factors of the ability to provide EOL care and the determination of the relationship between them can improve the provision of EOL care, reduction of health costs, timely and early referral of people to PC, and increase life quality.
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AIM: The aim of the study was to evaluate the prevalence of periodontitis in dentate people between 2011 and 2020. MATERIALS AND METHODS: PUBMED, Web of Science, and LILACS were searched up to and including December 2021. Epidemiological studies reporting the prevalence of periodontitis conducted between 2011 and 2020 were eligible for inclusion in this review. Studies were grouped according to the case definition of confidence as confident (Centers for Disease Control [CDC] AAP 2012; CDC/AAP 2007; and Armitage 1999) and non-confident (community periodontal index of 3 or 4, periodontal pocket depth >4 mm, and clinical attachment level ≥1 mm). Random effects meta-analyses with double arcsine transformation were conducted. Sensitivity subgroup and meta-regression analyses explored the effect of confounding variables on the overall estimates. RESULTS: A total 55 studies were included. The results showed a significant difference, with confident case definitions (61.6%) reporting nearly twice the prevalence as non-confident classifications (38.5%). Estimates using confident periodontal case definitions showed a pooled prevalence of periodontitis of 61.6%, comprising 17 different countries. Estimates reporting using the CDC/AAP 2012 case definition presented the highest estimate (68.1%) and the CDC/AAP 2007 presented the lowest (48.8%). Age was a relevant confounding variable, as older participants (≥65 years) had the highest pooled estimate (79.3%). CONCLUSION: Between 2011 and 2020, periodontitis in dentate adults was estimated to be around 62% and severe periodontitis 23.6%. These results show an unusually high prevalence of periodontitis compared to the previous estimates from 1990 to 2010.
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Periodontitis , Adulto , Humanos , Anciano , Prevalencia , Periodontitis/epidemiología , Estudios Epidemiológicos , Índice Periodontal , Bolsa PeriodontalRESUMEN
OBJECTIVES: Rheumatic immune-related adverse events (irAE) such as (poly)arthritis in patients undergoing immune checkpoint inhibitor (ICI) treatment pose a major clinical challenge. ICI therapy improves CD8+ T cell (CD8) function, but CD8 contributes to chronic inflammation in autoimmune arthritis (AA). Thus, we investigated whether immune functional and metabolic changes in CD8 explain the development of musculoskeletal irAE in ICI-treated patients. METHODS: Peripheral CD8 obtained from ICI-treated patients with and without arthritis irAEs and from AA patients with and without a history of malignancy were stimulated in media containing 13C-labelled glucose with and without tofacitinib or infliximab. Changes in metabolism, immune-mediator release, expression of effector cell-surface molecules and inhibition of tumour cell growth were quantified. RESULTS: CD8 from patients with irAE showed significantly lower frequency and expression of cell-surface molecule characteristic for activation, effector-functions, homing, exhaustion and apoptosis and reduced release of cytotoxic and proinflammatory immune mediators compared with CD8 from ICI patients who did not develop irAE. This was accompanied by a higher glycolytic rate and ATP production. Gene-expression analysis of pre-ICI-treated CD8 revealed several differentially expressed transcripts in patients who later developed arthritis irAEs. In vitro tofacitinib or infliximab treatment did not significantly change the immune-metabolic profile nor the capacity to release cytolytic mediators that inhibit the growth of the human lung cancer cell line H838. CONCLUSIONS: Our study shows that CD8 from ICI-treated patients who develop a musculoskeletal irAE has a distinct immune-effector and metabolic profile from those that remain irAE free. This specific irAE profile overlaps with the one observed in CD8 from AA patients and may prove useful for novel therapeutic strategies to manage ICI-induced irAEs.
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PURPOSE: De novo lipogenesis (DNL) is critical for cell growth and maintenance, and acetyl-CoA precursors can be derived from different substrates. We developed a 13 C NMR analysis of lipid extracts from cultured microglia cells administered with [U-13 C]glucose that informs overall lipogenic activity as well as the contribution of glucose to lipogenic acetyl-CoA. METHODS: BV-2 microglial cell line cultured with glucose and glutamine was provided with [U-13 C]glucose and unlabeled glutamine for 24 h and studied in either the presence or absence of lipopolysaccharide (LPS). Cells were then extracted for lipids and the crude lipid fraction was analyzed by 13 C NMR. 13 C-isotopomer signals in the fatty acid ω - 1 and ω - 2 signals representing consecutive or non-consecutive enrichment of the fatty acid chain by [1,2-13 C2 ]acetyl-CoA were quantified and applied to a probabilistic model of acetyl-CoA precursor and fatty acid enrichment. RESULTS: Glucose contributed 72 ± 2% of lipogenic acetyl-CoA while DNL from all sources accounted for 16 ± 2% of lipid turnover. With LPS, there was a significant decrease in glucose contribution (59 ± 4%, p < 0.05) while DNL was unchanged (11 ± 3%). CONCLUSIONS: A simple 13 C NMR analysis of the crude lipid fractions of BV-2 cells administered with [U-13 C]glucose informs DNL activity and the contribution of glucose to the acetyl-CoA precursors. While DNL was preserved in the presence of LPS, there was redirection of lipogenic acetyl-CoA sources from glucose to other substrates. Thus, in the present article, we describe a novel and simple 13 C NMR analysis approach to disclose the overall lipogenic activity and substrate contribution to DNL, suitable for evaluating DNL rates in cell cultures.
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Espectroscopía de Resonancia Magnética con Carbono-13/métodos , Lipogénesis , Microglía/metabolismo , Acetilcoenzima A/metabolismo , Animales , Células Cultivadas , Glucosa/metabolismo , RatonesRESUMEN
BACKGROUND AND OBJECTIVES: Randomized trials for mechanical thrombectomy (MT) excluded patients with ischemic strokes due to isolated posterior cerebral artery occlusion (IPCAO), and there is no evidence for best acute treatment strategy in these patients. We aimed to assess the effectiveness and safety of MT in acute IPCAO. METHODS: We retrospectively analyzed consecutive patients with acute stroke due to IPCAO submitted to MT and/or intravenous thrombolysis (IVT), between 2015-2019. Effectiveness outcomes (recanalization rate, first-pass effect, NIHSS 24h improvement and 3-month Modified Ranking Scale - mRS) and safety outcomes (complications, symptomatic intracranial hemorrhage (SICH) and 3-month mortality) were described and compared between groups. RESULTS: A total of 38 patients were included, 25 underwent MT and 13 had IVT alone. Successful and complete recanalization were achieved in 68% and 52% of MT patients, respectively. NIHSS improvement at 24h was found in 56% of MT patients versus 30.8% of patients submitted to IVT alone (OR [95% CI]=2.86 [0.69-11.82]) and excellent functional outcome at 3 months (mRS≤1) was achieved in 54.2% of MT patients versus 38.5% in the IVT group (OR [95% CI]=1.60 [0.41-6.32]). Complications occurred in 3 (12%) procedures and there were no SICH. Mortality at 3 months was 20% in the MT group and 15.4% in patients submitted to IVT alone. CONCLUSIONS: Our results reflect a real-world scenario in a single center and seem to support the recently growing literature showing that MT is a feasible and safe treatment in IPCAO, with favorable effectiveness.
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Arteriopatías Oclusivas , Trombolisis Mecánica , Arteria Cerebral Posterior , Enfermedad Aguda , Arteriopatías Oclusivas/complicaciones , Arteriopatías Oclusivas/terapia , Humanos , Trombolisis Mecánica/efectos adversos , Estudios Retrospectivos , Accidente Cerebrovascular/etiología , Resultado del TratamientoRESUMEN
A 59-year-old female patient was diagnosed with Whipple's disease (WD) after several months of constitutional complaints and adenopathies that were initially misinterpreted as sarcoidosis. Initial treatment included doxycycline, hydroxychloroquine and prednisolone, which was suspended due to long-term clinical stability. Four months after prednisolone suspension, the patient presented with right periorbital oedema and erythema. Ophthalmological examination revealed restricted eye movements. A computed tomography (CT) scan demonstrated signs of myositis. The patient was treated with anti-inflammatory and antibiotic drugs, that induced remission of the orbitopathy. During the following two years, she presented three relapses, affecting both the right or the left eyes. The last episode was also associated with systemic corticosteroid tapering. Orbitopathy is a rare form of WD presentation and the diagnosis of this condition may be challenging. As the clinical spectrum may range from an incidentaloma to a severe compressive neuropathy, proper recognition and management of WD orbitopathy is essential.
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Oftalmopatía de Graves , Enfermedad de Whipple , Antibacterianos/uso terapéutico , Doxiciclina , Femenino , Oftalmopatía de Graves/tratamiento farmacológico , Humanos , Persona de Mediana Edad , Tropheryma , Enfermedad de Whipple/diagnóstico , Enfermedad de Whipple/tratamiento farmacológicoRESUMEN
BACKGROUND: One of the most serious complications of diabetes mellitus (DM) is a diabetic foot ulcer (DFU), with lower extremity amputation (LEA). AIMS: This study aims to explore the role of anxiety and depression on mortality, reamputation and healing, after a LEA due to DFU. METHODS: A sample of 149 patients with DFU who underwent LEA answered the Hospital Anxiety and Depression Scale and a sociodemographic and clinical questionnaire. This is a longitudinal and multicenter study with four assessment moments that used Cox proportional hazards models adjusted for demographic and clinical variables. RESULTS: Rate of mortality, reamputation and healing, 10 months after LEA were 9.4%, 27.5% and 61.7%, respectively. Anxiety, at baseline, was negatively associated with healing. However, depression was not an independent predictor of mortality. None of the psychological factors was associated with reamputation. CONCLUSION: Results highlight the significant contribution of anxiety symptoms at pre-surgery, to healing after a LEA. Suggestions for psychological interventions are made.
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Amputación Quirúrgica , Pie Diabético , Humanos , Pie Diabético/cirugía , Ansiedad , Trastornos de Ansiedad , Factores de RiesgoRESUMEN
AIM: To evaluate the risk factors / predictors of tooth loss in patients with periodontitis who underwent periodontal therapy and long-term periodontal maintenance (PM). MATERIAL AND METHODS: PubMed, CENTRAL, EMBASE, Web of Science, LILACS and Scholar were searched up to and including September 2020. Studies limited to periodontitis patients who underwent active periodontal therapy (APT) and followed a regular PM programme with 5 years follow-up minimum were eligible for inclusion in this review. Studies were included if they reported data on tooth loss during PM. Random effects meta-analyses of number of tooth loss per patient per year were conducted. RESULTS: Thirty-six papers regarding thirty-three studies were included in this review, with three prospective 30 retrospective trials. Subgroup meta-analysis showed no differences between prospective and retrospective studies, with an average of 0.1 tooth loss per year per patient (p < 0.001). Maxillary and molar teeth were more susceptible to be extracted during long-term PM. Baseline characteristics (smoking, diabetes mellitus, cardiovascular disease, being male and teeth with furcation lesions) showed no significance as predictor of tooth loss through meta-regression. The percentage of tooth loss due to periodontal reasons ranged from 0.45% to 14.4%. The individual outcomes in each study evidenced different patient-related factors (age and smoking) and tooth-related factors (i.e. tooth type and location) were associated with tooth loss during PM. CONCLUSION: The majority of patients undergoing long-term PM have not lost teeth. On average, long-term PM effectively causes the loss of 1 tooth per patient every 10 years. Additional prospective trials may confirm these results.
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Periodontitis , Pérdida de Diente , Estudios de Seguimiento , Humanos , Masculino , Diente Molar , Estudios Prospectivos , Estudios Retrospectivos , Pérdida de Diente/epidemiología , Pérdida de Diente/etiologíaRESUMEN
Bile acids (BA) have shown promising effects in animal models of obesity. However, the said effects are thought to rely on a thermogenic effect, which is questionably present in humans. A previous work has shown that the BA chenodeoxycholic acid (CDCA) can revert obesity and accelerate metabolism in animal and cell culture models. Thus, the aim of this study was to understand if this obesity reduction is indeed thermogenically-dependent. A CRISPR/Cas9 model of TGR5 (BA receptor) knockdown in 3T3-L1 adipocytes was developed to diminish thermogenic effects. Various parameters were assessed, including mitochondrial bioenergetics by Seahorse flux analysis, oxidative stress and membrane potential by fluorometry, intermediary metabolism by NMR, protein content assessment by Western Blot, gene expression by qPCR, and confocal microscopy evaluation of mitophagy. CDCA was still capable, for the most part, of reversing the harmful effects of cellular obesity, elevating mitophagy and leading to the reduction of harmed mitochondria within the cells, boosting mitochondrial activity, and thus energy consumption. In summary, CDCA has a non-thermogenic, obesity reducing capacity that hinges on a healthy mitochondrial population, explaining at least some of these effects and opening avenues of human treatment for metabolic diseases.
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Fármacos Antiobesidad/farmacología , Sistemas CRISPR-Cas , Ácido Quenodesoxicólico/farmacología , Mitocondrias/metabolismo , Obesidad/tratamiento farmacológico , Receptores Acoplados a Proteínas G/deficiencia , Células 3T3-L1 , Animales , Técnicas de Silenciamiento del Gen , Ratones , Mitocondrias/genética , Obesidad/genética , Obesidad/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Termogénesis/efectos de los fármacos , Termogénesis/genéticaRESUMEN
Energy homeostasis is crucial for all physiological processes. Thus, when there is low energy intake, negative health effects may arise, including in reproductive function. We propose to study whether caloric restriction (CR) changes testicular metabolic profile and ultimately sperm quality. Male Wistar rats (n = 12) were randomized into a CR group fed with 30% fewer calories than weight-matched, ad libitum-fed animals (control group). Circulating hormonal profile, testicular glucagon-like peptide-1 (GLP-1), ghrelin and leptin receptors expression, and sperm parameters were analyzed. Testicular metabolite abundance and glycolysis-related enzymes were studied by NMR and Western blot, respectively. Oxidative stress markers were analyzed in testicular tissue and spermatozoa. Expressions of mitochondrial complexes and mitochondrial biogenesis in testes were determined. CR induced changes in body weight along with altered GLP-1, ghrelin, and leptin circulating levels. In testes, CR led to changes in receptor expression that followed those of the hormone levels; modified testicular metabolome, particularly amino acid content; and decreased oxidative stress-induced damage in testis and spermatozoa, although sperm head defects increased. In sum, CR induced changes in body weight, altering circulating hormonal profile and testicular metabolome and increasing sperm head defects. Ultimately, our data highlight that conditions of CR may compromise male fertility.
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Restricción Calórica , Ghrelina/metabolismo , Péptido 1 Similar al Glucagón/metabolismo , Leptina/metabolismo , Espermatozoides/metabolismo , Testículo/metabolismo , Animales , Western Blotting , Masculino , Metaboloma , Mitocondrias/metabolismo , Biogénesis de Organelos , Estrés Oxidativo , Espectroscopía de Protones por Resonancia Magnética , Distribución Aleatoria , Ratas , Ratas Wistar , Receptores de Leptina/metabolismo , Análisis de Semen , Cabeza del Espermatozoide/patología , Espermatozoides/patologíaRESUMEN
OBJECTIVES: The differential diagnosis of seronegative rheumatoid arthritis (negRA) and psoriasis arthritis (PsA) is often difficult due to the similarity of symptoms and the unavailability of reliable clinical markers. Since chronic inflammation induces major changes in the serum metabolome and lipidome, we tested whether differences in serum metabolites and lipids could aid in improving the differential diagnosis of these diseases. METHODS: Sera from negRA and PsA patients with established diagnosis were collected to build a biomarker-discovery cohort and a blinded validation cohort. Samples were analysed by proton nuclear magnetic resonance. Metabolite concentrations were calculated from the spectra and used to select the variables to build a multivariate diagnostic model. RESULTS: Univariate analysis demonstrated differences in serological concentrations of amino acids: alanine, threonine, leucine, phenylalanine and valine; organic compounds: acetate, creatine, lactate and choline; and lipid ratios L3/L1, L5/L1 and L6/L1, but yielded area under the curve (AUC) values lower than 70%, indicating poor specificity and sensitivity. A multivariate diagnostic model that included age, gender, the concentrations of alanine, succinate and creatine phosphate and the lipid ratios L2/L1, L5/L1 and L6/L1 improved the sensitivity and specificity of the diagnosis with an AUC of 84.5%. Using this biomarker model, 71% of patients from a blinded validation cohort were correctly classified. CONCLUSIONS: PsA and negRA have distinct serum metabolomic and lipidomic signatures that can be used as biomarkers to discriminate between them. After validation in larger multiethnic cohorts this diagnostic model may become a valuable tool for a definite diagnosis of negRA or PsA patients.
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Artritis Psoriásica/sangre , Artritis Reumatoide/sangre , Acetatos/sangre , Adulto , Anciano , Anciano de 80 o más Años , Alanina/sangre , Aminoácidos/sangre , Artritis Psoriásica/diagnóstico , Artritis Reumatoide/diagnóstico , Artritis Reumatoide/inmunología , Colina/sangre , Creatina/sangre , Diagnóstico Diferencial , Femenino , Humanos , Ácido Láctico/sangre , Lipidómica , Lípidos/sangre , Masculino , Metaboloma , Metabolómica , Persona de Mediana Edad , Fosfocreatina/sangre , Espectroscopía de Protones por Resonancia Magnética , Ácido Succínico/sangreRESUMEN
Mitoxantrone (MTX) is used to treat several types of cancers and to improve neurological disability in multiple sclerosis. Unfortunately, cardiotoxicity is a severe and common adverse effect in MTX-treated patients. Herein, we aimed to study early and late mechanisms of MTX-induced cardiotoxicity using murine HL-1 cardiomyocytes. Cells were exposed to MTX (0.1, 1 or 10 µM) during short (2, 4, 6, or 12 h) or longer incubation periods (24 or 48 h). At earlier time points, (6 and 12 h) cytotoxicity was already observed for 1 and 10 µM MTX. Proteomic analysis of total protein extracts found 14 proteins with higher expression and 26 with lower expression in the cells exposed for 12 h to MTX (pH gradients 4-7 and 6-11). Of note, the expression of the regulatory protein 14-3-3 protein epsilon was increased by a factor of two and three, after exposure to 1 and 10 µM MTX, respectively. At earlier time-points, 10 µM MTX increased intracellular ATP levels, while decreasing media lactate levels. At later stages (24 and 48 h), MTX-induced cytotoxicity was concentration and time-dependent, according to the MTT reduction and lactate dehydrogenase leakage assays, while caspase-9, -8 and -3 activities increased at 24 h. Regarding cellular redox status, total glutathione increased in 1 µM MTX (24 h), and that increase was dependent on gamma-glutamylcysteine synthetase activity. Meanwhile, for both 1 and 10 µM MTX, oxidized glutathione was significantly higher than control at 48 h. Moreover, MTX was able to significantly decrease proteasomal chymotrypsin-like activity in a concentration and time-independent manner. In summary, MTX significantly altered proteomic, energetic and oxidative stress homeostasis in cardiomyocytes at clinically relevant concentrations and our data clearly demonstrate that MTX causes early cardiotoxicity that needs further study.
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Metabolismo Energético/efectos de los fármacos , Cardiopatías/inducido químicamente , Mitoxantrona/toxicidad , Miocitos Cardíacos/efectos de los fármacos , Complejo de la Endopetidasa Proteasomal/metabolismo , Proteoma , Proteómica , Animales , Proteínas Reguladoras de la Apoptosis/metabolismo , Cardiotoxicidad , Línea Celular , Relación Dosis-Respuesta a Droga , Cardiopatías/metabolismo , Ratones , Miocitos Cardíacos/metabolismo , Estrés Oxidativo/efectos de los fármacos , Carbonilación Proteica , Factores de TiempoRESUMEN
BACKGROUND: Mechanical thrombectomy is the standard treatment in acute ischemic stroke due to large vessel occlusion, but there is limited evidence about its efficacy in very old patients. We sought to analyse safety and effectiveness of mechanical thrombectomy in nonagenarian versus octagenarian patients. METHODS: We included consecutive patients with acute ischemic stroke due to large vessel occlusion subjected to mechanical thrombectomy, during 29 months in a tertiary center. Patients were divided into two sub-groups, according to age: 80-89 and >90 years old. Recanalization, complications, functional outcome and mortality at discharge and at 3 months were compared. Multivariable analysis was performed to identify independent predictors of functional outcome at 3 months of follow-up, assessed by the modified Rankin Scale. RESULTS: A total of 128 octogenarians (88.9%) and 16 nonagenarians (11.1%) met the inclusion criteria. Successful revascularization was achieved in 87.5% of octagenarians and in 81.3% of nonagenarians (pâ¯=â¯0.486). Symptomatic hemorrhage occurred in 3.1% and 6.3% of younger and older patients, respectively (pâ¯=â¯0.520). Cerebral edema occured in 35.2% of octagenarians versus 25.0% of nonagenarians (pâ¯=â¯0.419). Functional independence (mRS ≤ 2) at 3 months was achieved in 28 (22.6%) and 5 (31.3%) of octagenarians and nonagenarians, respectively (pâ¯=â¯0.445). Mortality at 3 months was not significantly higher in nonagenarians (37.5%) versus octagenarians (33.9%, pâ¯=â¯0.773). CONCLUSIONS: No significant diferences were found in functional outcome, mortality, recanalization and complication rates between octagenarians and nonagenarians submitted to mechanical thrombectomy, underlining that patients should not be excluded from mechanical thrombectomy based on age alone.
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Isquemia Encefálica/terapia , Accidente Cerebrovascular/terapia , Trombectomía , Factores de Edad , Anciano de 80 o más Años , Isquemia Encefálica/diagnóstico , Isquemia Encefálica/mortalidad , Isquemia Encefálica/fisiopatología , Toma de Decisiones Clínicas , Evaluación de la Discapacidad , Femenino , Mortalidad Hospitalaria , Humanos , Masculino , Alta del Paciente , Selección de Paciente , Recuperación de la Función , Estudios Retrospectivos , Medición de Riesgo , Factores de Riesgo , Accidente Cerebrovascular/diagnóstico , Accidente Cerebrovascular/mortalidad , Accidente Cerebrovascular/fisiopatología , Trombectomía/efectos adversos , Trombectomía/mortalidad , Factores de Tiempo , Resultado del TratamientoRESUMEN
Obesity incidence has pandemic proportions and is expected to increase even further. Glucagon-like peptide-1 (GLP-1) based therapies are well-established pharmacological resources for obesity treatment. GLP-1 regulates energy and glucose homeostasis, which are also crucial for spermatogenesis. Herein, we studied the GLP-1 effects in human Sertoli cells (hSCs) metabolism and mitochondrial function. hSCs were cultured in absence or exposed to increasing doses of GLP-1 mimicking physiological post-prandial (0.01â¯nM) levels or equivalent to pharmacological levels (1 and 100â¯nM) used for obesity treatment. We identified GLP-1 receptor in hSCs. Consumption/production of extracellular metabolites were assessed, as well as protein levels or activities of glycolysis-related enzymes and transporters. Mitochondrial membrane potential and oxidative damage were evaluated. Glucose consumption decreased, while lactate production increased in hSCs exposed to 0.01 and 1â¯nM GLP-1. Though lactate dehydrogenase (LDH) protein decreased after exposure to 100â¯nM GLP-1 its activity increased in hSCs exposed to the same concentration of GLP-1. Mitochondrial membrane potential decreased in hSCs exposed to 100â¯nM of GLP-1, while formation of carbonyl groups was decreased in those cells. Those effects were followed by an increase in p-mammalian target of rapamycin (mTOR) Ser(2448). Overall, the lowest concentrations of GLP-1 increased the efficiency of glucose conversion to lactate, while GLP-1 concentration of 100â¯nM induces mTOR phosphorylation, decreases mitochondrial membrane potential and oxidative damage. GLP-1 regulates testicular energy homeostasis and pharmacological use of GLP-1 analogues could be valuable to counteract the negative impact of obesity in male reproductive function.
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Péptido 1 Similar al Glucagón/farmacología , Células de Sertoli/efectos de los fármacos , Células Cultivadas , Metabolismo Energético/efectos de los fármacos , Receptor del Péptido 1 Similar al Glucagón/fisiología , Glucosa/metabolismo , Humanos , Ácido Láctico/metabolismo , Masculino , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Células de Sertoli/fisiologíaRESUMEN
INTRODUCTION: Feed optimization is a key step to the environmental and economic sustainability of aquaculture, especially for carnivorous species. Plant-derived ingredients can contribute to reduce costs and nitrogenous effluents while sparing wild fish stocks. However, the metabolic use of carbohydrates from vegetable sources by carnivorous fish is still not completely understood. OBJECTIVES: We aimed to study the effects of diets with carbohydrates of different digestibilities, gelatinized starch (DS) and raw starch (RS), in the muscle metabolome of European seabass (Dicentrarchus labrax). METHODS: We followed an NMR-metabolomics approach, using two sample preparation procedures, the intact muscle (HRMAS) and the aqueous muscle extracts (1H NMR), to compare the variations in muscle metabolome between the two diets. RESULTS: In muscle, multivariate analysis revealed similar metabolome shifts for DS and RS diets, when compared with the control diet. HRMAS of intact muscle, which included both hydrophobic and hydrophilic metabolites, showed increased lipid in DS-fed fish by univariate analysis. Regardless of the nature of the starch, increased glycine and phenylalanine, and decreased proline were observed when compared to the Ctr diet. Combined univariate analysis of intact muscle and aqueous extracts indicated specific diet related changes in lipid and amino acid metabolism, consistent with increased dietary carbohydrate supplementation. CONCLUSIONS: Due to differential sample processing, outputs differ in detail but provide complementary information. After tracing nutritional alterations by profiling fillet components, DS seems to be the most promising alternative to fishmeal-based diets in aquaculture. This approach should be reproducible for other farmed fish species and provide valuable information on nutritional and organoleptic properties of the final product.