Study of the relationship of bacterial metabolites Trimethylamine N-oxide and Trimethylamine levels with cardiovascular disease and renal function in HIV-infected patients

Tesis de la Universidad Complutense de Madrid, Facultad de Medicina, Departamento de Medicina, leída el 23/11/2018Los avances en la terapia antirretroviral para el tratamiento de la infección por VIH han supuesto un antes y un después en el manejo clínico y la calidad de vida de los pacientes, consiguiendo prolongadas supervivencias libres de enfermedad en la mayor parte de los pacientes que viven con VIH (PWIH) (Duprez, 2009). Actualmente la enfermedad cardiovascular {ECV) es una de las principales causas de mortalidad en PWIH presentando mayor riesgo que la población general. Los factores de riesgo tradicionales son responsables de una parte importante pero no son los únicos, también lo son la inflamación crónica causada por el VIH, la inmunodeficiencia y la posible acción directa de ciertos antirretrovirales (GEAM, 2014). Es conocido que algunos metabolitos generados por la microbiota intestinal son proaterogénicos y que en PWIH, incluyendo a los que controlan la replicación viral con TAR, la microbiota intestinal es muy distinta de la población no infectada. La disbiosis constante puede producir un impacto en la actividad inmunológica del intestino originando la difusión de bacterias e inflamación sistémica (Ufnal, 2015). Los metabolitos de la fosfatidilcolina en la dieta son esenciales para ciertas funciones biológicas pero también parece que poseen cierto potencial aterogénico. La fosfatidilcolina dietética es una fuente importante de colina, que se metaboliza por la microbiota intestinal a trimetilamina {TMA). En el hígado, la TMA se oxida a trimetilamina-N-óxido (TMAO) (Tang, 2014; Ufnal, 2015). La evidencia generada en población no VIH durante los últimos años sugiere que concentraciones plasmáticas de TMAO elevadas podrían tener un papel como un nuevo marcador de un mayor riesgo cardiovascular. Estudios clínicos apuntan hacia una correlación positiva entre elevados niveles plasmáticos de TMAO, no con el TMA, y un aumento del riesgo cardiovascular, independientemente de los factores de riesgo tradicionales. Una baja tasa de filtrado glomerular junto a otros factores de riesgo, como son la hipertensión y la diabetes mellitus, fueron habituales en el cuartil superior de niveles plasmáticos de TMAO. (Tang, 2014; Ufnal, 2015). Los niveles de TMAO también se han relacionado con enfermedades renales. Concentraciones elevadas de TMAO se han asociado con una disminución de la función renal en pacientes con ERC normalizándose tras el trasplante renal. Los niveles de TMAO correlacionaron un aumento de la inflamación sistémica y fueron un predictor independiente de la mortalidad en pacientes con ERC 3-5 (Missailidis, 2016)...The advances in antiretroviral therapy for HIV infection have been a befare and after in the clinical management and quality of life of HIV patients, achieving prolongad disease-free survival (Duprez , 2009). Currently, cardiovascular disease (CD) is one of the main causes of mortality in PLHIV, presenting a higher risk than general population. The traditional risk factors are responsible for a relevant part but are not the only ones, also are the chronicinflammation caused by HIV, immunodeficiency and the possible direct action of certain antiretroviral drugs (GEAM, 2014). lt is known that sorne metabolites generated by the intestinal microbiota are proatherogenic and that in PLHIV the intestinal microbiota is very different from the non-infectad population, including those patients well controlled with TAR. Continuous dysbiosis can produce an impact on the immunological activity of the intestina, causing bacteria! diffusion and systemic inflammation (Ufnal, 2015). Metabolites of diet phosphatidylcholine are essential for certain biological functions but also appear to have certain atherogenic activity. Dietary phosphatidylcholine is an important source of choline, which is metabolized by intestinal microbiota to trimethylamine (TMA). In the liver, the TMA is oxidized to trimethylamine-N-oxide (TMAO) (Tang, 2014; Ufnal, 2015). The evidence generated in the non-HIV population during the last years suggests that high plasma concentration of TMAO may have a role as a new marker of increased cardiovascular risk. Clinical trials indicate a positiva correlation between elevated plasma TMAO levels, not TMA, and increased cardiovascular risk, regardless of traditional risk factors. A low glomerular filtration rate together with other risk factors, such as hypertension and diabetes mellitus, were common in the highest quartile of plasma TMAO levels (Tang, 2014, Ufnal, 2015). Variations in TMAO levels have also been relatad to kidney disease. Elevated TMAO levels were strongly associated with decrease of renal function in CKD and were normaliza after renal transplantation. TMAO levels correlatas with increased systemic inflammation and is an independent predictor of mortality in CKD 3-5 patients (Missailidis, 2016)...Depto. de MedicinaFac. de MedicinaTRUEunpu

Novel Oleanolic and Maslinic Acids derivatives as a promising treatment against bacterial biofilm in nosocomial infections: An in Vitro and in Vivo study.

Oleanolic acid (OA) and maslinic acid (MA) are pentacyclic triterpenic compounds that abound in industrial olive oil waste. These compounds have renowned antimicrobial properties and lack cytotoxicity in eukaryotic cells as well as resistance mechanisms in bacteria. Despite these advantages, their antimicrobial activity has only been tested in vitro, and derivatives improving this activity have not been reported. In this work, a set of 14 OA and MA C-28 amide derivatives have been synthesized. Two of these derivatives, MA-HDA and OA-HDA, increase the in vitro antimicrobial activity of the parent compounds while reducing their toxicity in most of the Gram-positive bacteria tested, including a methicillin-resistant Staphylococcus aureus-MRSA. MA-HDA also shows an enhanced in vivo efficacy in a Galleria mellonella invertebrate animal model of infection. A preliminary attempt to elucidate their mechanism of action revealed that these compounds are able to penetrate and damage the bacterial cell membrane. More significantly, their capacity to reduce antibiofilm formation in catheters has also been demonstrated in two sets of conditions: a static and a more challenged continuous-flow S. aureus biofilm.This study was partially supported by grants from the Ministerio de Economiá , Industria y Competitividad, MINECO, and Agencia Estatal de Investigación, AEI, Spain, cofunded by Fondo Europeo de Desarrollo Regional, FEDER, European Union (BIO2015−63557-R, FIS2017−85954-R and RTI2018−098573-B-100)

Different associations of intentional and non-intentional non-adherence behaviors with patient experience with healthcare and patient beliefs in medications: a survey of patients with chronic conditions

[Abstract] Purpose: To investigate relationships between intentional and non-intentional non-adherence behaviors and patient experience with healthcare and beliefs in medications. Patients and methods: This is a post hoc analysis of a cross-sectional anonymous survey distributed between May and September 2017 to patients with rheumatic disease, inflammatory bowel disease, HIV infection or diabetes mellitus from outpatient and primary care clinics in Spain. Patients answered five questions about non-adherence behaviors and completed questionnaires on their experience with healthcare (IEXPAC: Instrument to Evaluate the EXperience of PAtients with Chronic diseases) and beliefs about medicines (BMQ: Beliefs About Medicines Questionnaire). Results: Among 1530 respondents, 53% showed ≥1 non-adherence behavior; 35% had ≥1 non-intentional non-adherence behavior, and 33% had ≥1 intentional non-adherence behavior. Patients with HIV infection had the lowest frequency of intentional non-adherence behaviors. Non-intentional non-adherence was associated with patient beliefs (inversely with BMQ overall score) and patient experiences (inversely with IEXPAC Factor 3 sub-score, self-management). Intentional non-adherence was strongly associated with beliefs scores (directly with BMQ concerns and inversely with BMQ necessity sub-score) and inversely associated with HIV infection. Conclusion: The different associations of intentional and non-intentional non-adherence behaviors found in this study help to understand how patient experiences and beliefs influence medical non-adherence, and in the development of strategies for reducing non-adherence

The Influence of Patient Experience with Healthcare on the Health-Related Quality of Life of People Living with HIV: An Observational Cross-Sectional Survey

[Abstract] Introduction Patient experience is central to the quality of healthcare delivery, showing positive associations with several outcome measures. The main objectives of this study are to analyze the influence of patient experience on the health-related quality of life in people living with HIV and the role played by treatment complexity and clinical care. Methods We conducted a cross-sectional survey with 467 patients with HIV. We used the Instrument for Evaluation of the Experience of Chronic Patients and the Health-related Quality of Life Questionnaire (EQ-5D-5L). We analyzed a predictive model through the partial least squares (PLS) method. Results The patient self-management scores showed the highest positive relationship with the patient’s health-related quality of life (β = 0.24, β = 0.32, p  50 years old (p < 0.05). Conclusions Patient experience mainly influenced the health-related quality of life of older people living with HIV. The treatment and clinical care complexity played an important role in degrading the patients' experience and their quality of life. More integrated care would benefit the health-related quality of life of people living with HIV

N-ACETYLCYSTEINE DOES NOT IMPROVE SPERM MOTILITY OF LIDIA BULL AFTER PROLONGED EPIDIDYMAL STORAGE / LA N-ACETILCISTEINA NO MEJORA LA MOTILIDAD ESPERMÁTICA EN TOROS DE LIDIA LUEGO DEL ALMACENAMIENTO EPIDIDIMARIO PROLONGADO

The Lidia bovine breed is considered a hallmark of Spanish cattle industry. Assisted reproductive techniques like cryopreservation of epididymal spermatozoa could be considered as an important tool to obtain more offspring and store its genetics. As these bulls are not selected by their reproductive performance or sperm freezability, the quality of their ejaculates is poor and addition of antioxidants prior cryopreservation could exert beneficial effects on the post-thaw sperm quality. The aim of this study was to evaluate the effect of the supplementing a tris-fructose-egg yolk based freezing extender with1 mMand2.5 mMof N-acetylcysteine to sperm recovered from epididymis stored at4ºCfor 24, 48, 72 or 96 hours prior cryopreservation. Motility values and sperm kinematic parameters were compared against control (epididymis stored for 24 hours and no antioxidant addition). Our results showed that N-acetylcysteine addition did not improve sperm motility parameters at any of the time points or dosages tested. In addition, storage of bullfight epididymis up to 96 hours did not significantly affect sperm kinematic parameters or total and progressive motility.RESUMENLa raza de Lidia es considerada una insignia de la industria ganadera española. Las tecnologías de reproducción asistida como la criopreservación de espermatozoides epididimarios podría ser considerara como una herramienta importante para obtener más crías y conservar su genética. Ya que estos toros no son seleccionados por su desempeño reproductivo o congelabilidad espermática, la calidad de sus eyaculados es pobre y la adición de antioxidantes antes de la criopreservación podría tener efectos beneficiosos sobre la calidad del semen descongelado. El propósito de este estudio fue evaluar el efecto de la suplementación del medio a base de tris-fructosa y yema de huevo con 1 mM y 2,5 mM de N-acetilcisteína a espermatozoides recuperados de epidídimos almacenados a 4°C por 24, 48, 72 o 96 horas antes de la criopreservación. Los valores de movilidad y los parámetros de cinética espermática fueron comparados con el control (epidídimos almacenados por 24 horas y sin la adición de antioxidantes). Nuestros resultados muestran que la adición de N-acetilcisteína no mejora los parámetros de motilidad espermática en ninguno de los momentos o dosis evaluadas. Además, el almacenamiento de epidídimos refrigerados hasta 96 horas no afecta significativamente los parámetros de cinética espermática o la movilidad total o progresiva.

Outlining adequate protocols for Lidia bull epididymal storage and sperm cryopreservation: use of glycerol, dimethylformamide and N-acetylcysteine

P. 1-10The Lidia bovine breed is an important hallmark of the Spanish cattle industry. Bulls are selected based upon aggressiveness and epididymal sperm cryopreservation is the way to obtain and store their genetics. There are not specifically designed protocols yet to perform Lidia bull sperm cryopreservation. The present study aimed to determine if a tris-fructose-citrate-egg yolk (20% v/v; TFY) extender supplemented with 7% glycerol (TFY1) or 3.5% glycerol plus 3.5% dimethylformamide (DMF; TFY2) are suitable media for cryopreservation of epididymal Lidia bull sperm. Moreover, the effect of N-acetylcysteine (NAC), a potent antioxidant, was evaluated. The epididymis were stored at 4°C for 24, 48, 72 or 96 h, and both freezing media were tested as such or supplemented with 1 or 2.5 mM of NAC. Our data demonstrated that post-thaw viability was well maintained (TFY1: 50.8% ± 1.9 at 24 h and 52.4% ± 0.8 at 96 h and TFY2: 52.6% ± 1.6 at 24 h and 56.1% ± 1.8 at 96 h; mean % ± SEM; p>0.05) as also were total and progressive sperm motility, high mitochondrial membrane potential, ROS production, DNA status and acrosomal intactness of Lidia bull sperm up to 96 h of epididymal storage, all extender variations being similar (p>0.05). In conclusion, the use of TFY medium supplemented either with 7% glycerol alone or the combination of 3.5% glycerol and 3.5% DMF were equally safe choices for epididymal Lidia bull sperm cryopreservation, and NAC addition did not significantly improve sperm post-thaw quality.S

Outlining adequate protocols for Lidia bull epididymal storage and sperm cry-opreservation: use of glycerol, dimethylformamide and N-acetylcysteine

La raza bovina Lidia es un sello distintivo importante de la industria ganadera española. Los toros se seleccionan en función de la agresividad y la crioconservación de espermatozoides epididimales es la forma de obtener y almacenar su genética. Todavía no hay protocolos diseñados específicamente para realizar la crioconservación de espermatozoides Lidia bull. El presente estudio tuvo como objetivo determinar si un medio extensor de tris-fructosa-citrato-huevo (20% v / v; TFY) suplementado con 7% de glicerol (TFY1) o 3,5% de glicerol más 3,5% de dimetilformamida (DMF; TFY2) son medios adecuados Para la crioconservación de epididimales Lidia esperma de toro. Además, se evaluó el efecto de la N-acetilcisteína (NAC), un potente antioxidante. Los epidídimos se almacenaron a 4 ° C durante 24, 48, 72 o 96 h, y ambos medios de congelación se probaron como tales o se complementaron con 1 o 2,5 mM de NAC. Nuestros datos demostraron que la viabilidad posterior a la descongelación se mantuvo bien (TFY1: 50.8% ± 1.9 a las 24 h y 52.4% ± 0.8 a las 96 hy TFY2: 52.6% ± 1.6 a las 24 h y 56.1% ± 1.8 a las 96 h; media% ± SEM; p> 0.05) al igual que la motilidad espermática total y progresiva, el potencial de membrana mitocondrial alto, la producción de ROS, el estado del ADN y la intactidad acrosomal del esperma de toro Lidia hasta 96 h de almacenamiento de epidídimo, todas las variaciones del extensor son similares (p> 0.05 ). En conclusión, el uso de medio TFY suplementado con un 7% de glicerol solo o la combinación de un 3,5% de glicerol y un 3,5% de DMF fue una elección igualmente segura para la crioconservación de espermatozoides Lidia epididimales, y la adición de NAC no mejoró significativamente la calidad del esperma después del deshielo.The Lidia bovine breed is an important hallmark of the Spanish cattle industry. Bulls are selected based upon aggressiveness and epididymal sperm cryopreservation is the way to obtain and store their genetics. There are not specifically designed protocols yet to perform Lidia bull sperm cryopreservation. The present study aimed to determine if a tris-fructose-citrate-egg yolk (20% v/v; TFY) extender supplemented with 7% glycerol (TFY1) or 3.5% glycerol plus 3.5% dimethylformamide (DMF; TFY2) are suitable media for cryopreservation of epididymal Lidia bull sperm. Moreover, the effect of N-acetylcysteine (NAC), a potent antioxidant, was evaluated. The epididymis were stored at 4°C for 24, 48, 72 or 96 h, and both freezing media were tested as such or supplemented with 1 or 2.5 mM of NAC. Our data demonstrated that post-thaw viability was well maintained (TFY1: 50.8% ± 1.9 at 24 h and 52.4% ± 0.8 at 96 h and TFY2: 52.6% ± 1.6 at 24 h and 56.1% ± 1.8 at 96 h; mean % ± SEM; p>0.05) as also were total and progressive sperm motility, high mitochondrial membrane potential, ROS production, DNA status and acrosomal intactness of Lidia bull sperm up to 96 h of epididymal storage, all extender variations being similar (p>0.05). In conclusion, the use of TFY medium supplemented either with 7% glycerol alone or the combination of 3.5% glycerol and 3.5% DMF were equally safe choices for epididymal Lidia bull sperm cryopreservation, and NAC addition did not significantly improve sperm post-thaw quality.• Ministerio de Economía, Industria y Competitividad. Proyecto ITC-2015-1342 • Ministerio de Economía y Competitividad. Beca Juan de la Cierva IJCI- 2014-19428, para Beatriz Macías García • Centro de Cirugía de Mínima Invasión Jesús Usón. Beca para Elvira Matilla Pinto • Ministerio de Economía, Industria y Competitividad y Fondos FEDER. Proyecto AGL2015-73249-JIN (I+D+i) • Fundação para a Ciência e a Tecnologia (Portugal)/ESF/Ministerio de Ciencia, Tecnología y Educación Superior. Beca SFRH/BPD/85532/2012, para Lauro González Fernández • Ministerio de Educación, Cultura y Deportes. Beca FPU-014/03449, para Violeta Calle GuisadopeerReviewe

Healthcare experience and their relationship with demographic, disease and healthcare-related variables: a cross-sectional survey of patients with chronic diseases using the IEXPAC scale.

[Abstract] BACKGROUND: Patient experience is acknowledged as a principal aspect of quality healthcare delivery, and it has implications with regard to outcomes. OBJECTIVES: Our objective was to evaluate the healthcare experience of patients with chronic diseases to identify patient-perceived healthcare gaps and to assess the influence of demographic and healthcare-related variables on patient experiences. METHODS: A cross-sectional survey was delivered to adult patients with chronic diseases: diabetes mellitus (DM), human immunodeficiency virus (HIV) infection, inflammatory bowel disease (IBD) or rheumatic diseases. Patient experiences were assessed with the Instrument for Evaluation of the Experience of Chronic Patients (IEXPAC) questionnaire, with possible scores ranging from 0 (worst) to 10 (best experience). RESULTS: Of the 2474 patients handed the survey, 1618 returned it (response rate 65.4%). Patients identified gaps in healthcare related mainly to access to reliable information and services, interaction with other patients and continuity of healthcare after hospital discharge. The mean ± standard deviation (SD) IEXPAC score was 6.0 ± 1.9 and was higher for patients with HIV (6.6 ± 1.7) than for those with rheumatic disease (5.5 ± 2.0), IBD (5.9 ± 2.0) or DM (5.9 ± 1.9) (p < 0.001). In multivariate models, better overall IEXPAC experience was associated with follow-up by the same physician, follow-up by a nurse, receiving healthcare support from others and treatment with subcutaneous or intravenous drugs. The multivariate model that confirmed patients with HIV or DM had better experience than did those with rheumatic diseases. CONCLUSIONS: Through IEXPAC, patients identified aspects for healthcare quality improvements and circumstances associated with better experience, which may permit greater redirection of healthcare toward patient-centered goals while facilitating improvements in social care and long-term healthcare quality

Fecal Metabolome and Bacterial Composition in Severe Obesity:Impact of Diet and Bariatric Surgery

The aim of this study was to monitor the impact of a preoperative low-calorie diet and bariatric surgery on the bacterial gut microbiota composition and functionality in severe obesity and to compare sleeve gastrectomy (SG) versus Roux-en-Y gastric bypass (RYGB). The study also aimed to incorporate big data analysis for the omics results and machine learning by a Lasso-based analysis to detect the potential markers for excess weight loss. Forty patients who underwent bariatric surgery were recruited (14 underwent SG, and 26 underwent RYGB). Each participant contributed 4 fecal samples (baseline, post-diet, 1 month after surgery and 3 months after surgery). The bacterial composition was determined by 16S rDNA massive sequencing using MiSeq (Illumina). Metabolic signatures associated to fecal concentrations of short-chain fatty acids, amino acids, biogenic amines, gamma-aminobutyric acid and ammonium were determined by gas and liquid chromatography. Orange 3 software was employed to correlate the variables, and a Lasso analysis was employed to predict the weight loss at the baseline samples. A correlation between Bacillota (formerly Firmicutes) abundance and excess weight was observed only for the highest body mass indexes. The low-calorie diet had little impact on composition and targeted metabolic activity. RYGB had a deeper impact on bacterial composition and putrefactive metabolism than SG, although the excess weight loss was comparable in the two groups. Significantly higher ammonium concentrations were detected in the feces of the RYGB group. We detected individual signatures of composition and functionality, rather than a gut microbiota characteristic of severe obesity, with opposing tendencies for almost all measured variables in the two surgical approaches. The gut microbiota of the baseline samples was not useful for predicting excess weight loss after the bariatric process

PiRNA-associated proteins and retrotransposons are differentially expressed in murine testis and ovary of aryl hydrocarbon receptor deficient mice

Previous studies suggested that the aryl hydrocarbon receptor (AhR) contributes to mice reproduction and fertility. However, the mechanisms involved remain mostly unknown. Retrotransposon silencing by Piwi-interacting RNAs (piRNAs) is essential for germ cell maturation and, remarkably, AhR has been identified as a regulator of murine B1-SINE retrotransposons. Here, using littermate AhR+/+ and AhR-/- mice, we report that AhR regulates the general course of spermatogenesis and oogenesis by a mechanism likely to be associated with piRNA-associated proteins, piRNAs and retrotransposons. piRNA-associated proteins MVH and Miwi are upregulated in leptotene to pachytene spermatocytes with a more precocious timing in AhR-/- than in AhR+/+ testes. piRNAs and transcripts from B1-SINE, LINE-1 and IAP retrotransposons increased at these meiotic stages in AhR-null testes. Moreover, B1-SINE transcripts colocalize with MVH and Miwi in leptonema and pachynema spermatocytes. Unexpectedly, AhR-/- males have increased sperm counts, higher sperm functionality and enhanced fertility than AhR+/+ mice. In contrast, piRNA-associated proteins and B1-SINE and IAP-derived transcripts are reduced in adult AhR-/- ovaries. Accordingly, AhR-null female mice have lower numbers of follicles when compared with AhR+/+ mice. Thus, AhR deficiency differentially affects testis and ovary development possibly by a process involving piRNA-associated proteins, piRNAs and transposable elements.Trabajo financiado por: Ministerio de Ciencia e Innovación. Ayuda BFU2011-22678 para Pedro María Fernández Salguero Ministerio de Economía y Competitividad. Ayuda SAF2014-51813-R para Pedro María Fernández Salguero Junta de Extremadura. Ayuda GR15008, para Pedro María Fernández Salguero Red Temática de Investigación Cooperativa en Cáncer (RTICC), Instituto Carlos III y Ministerio de Economía y Competitividad (RD12/0036/0032). Trabajos de laboratorio de Pedro María Fernández Salguero Ministerio de Economía y Competitividad. Ayuda AGL2013-43211-R, para Fernando Juan Peña Vega Red Temática de Investigación Cooperativa en Cáncer (RTICC), Instituto Carlos III. Ayuda para Eva María Rico Leo Ministerio de Economía y Competitividad. Ayuda para Francisco Javier González Rico Junta de Extremadura. Ayuda para Eva María Barrasa Ardila Ministerio de Educación, Cultura y Deportes. Beca de Formación de Personal de Investigación, para Nuria Moreno Marín Ministerio de Educación, Cultura y Deportes. Beca FPU13/03991, de Formación de Profesorado Universitario, para Patricia Martín Muñoz Ministerio de Economía y Competitividad. Beca Juan de la Cierva IJCI-2014-21671, para Cristina Ortega Ferrusola Ministerio de Economía y Competitividad, Beca BFU2014-59307-R, para Alberto M. Pendás y Elena Llano Cuadro MEIONet, Junta de Castilla y León y el Programa de Fondeos FEDER, de la Unión EuropeapeerReviewe