The Proof-of-Concept of MBA121, a Tacrine-Ferulic Acid Hybrid, for Alzheimer's Disease Therapy.

Great effort has been devoted to the synthesis of novel multi-target directed tacrine derivatives in the search of new treatments for Alzheimer's disease (AD). Herein we describe the proof of concept of MBA121, a compound designed as a tacrine-ferulic acid hybrid, and its potential use in the therapy of AD. MBA121 shows good ß-amyloid (Aß) anti-aggregation properties, selective inhibition of human butyrylcholinesterase, good neuroprotection against toxic insults, such as Aß1-40, Aß1-42, and H2O2, and promising ADMET properties that support translational developments. A passive avoidance task in mice with experimentally induced amnesia was carried out, MBA121 being able to significantly decrease scopolamine-induced learning deficits. In addition, MBA121 reduced the Aß plaque burden in the cerebral cortex and hippocampus in APPswe/PS1ΔE9 transgenic male mice. Our in vivo results relate its bioavailability with the therapeutic response, demonstrating that MBA121 is a promising agent to treat the cognitive decline and neurodegeneration underlying AD.

Genotype FBN1/phenotype relationship in a cohort of patients with Marfan syndrome.

Marfan syndrome (MFS) is a systemic connective tissue disorder caused by mutations in the fibrillin-1 (FBN1) gene, and cardiovascular involvement is the leading cause of mortality. We sought to examine the genotype/phenotype realtionship in 61 consecutive patients with a phenotype and genotype compatible with MFS. The FBN1 gene was analyzed by massive sequencing using a hybridization capture-based target enrichment custom panel. Forty-three different variants of FBN1 were identified, of which 17 have not been previously reported. The causal variants of MFS were grouped into mutations resulting in haploinsufficiency (HI group; 23 patients) and mutations producing a dominant-negative effect (DN group; 38 patients). Patient information was collected from electronic medical records and clinical evaluation. While no significant differences were found between the two groups, the HI group included more cases with aortic dissection and occurring at a younger age that the DN group (34.7% vs. 15.8%; p = 0.160). Irrespective of the mutation group, males presented with a higher probability of aortic involvement (4-fold higher risk than females) and aortic dissections events occurred at younger ages. Patients with DN variants carrying a cysteine substitution had a higher incidence of ectopia lentis.

A dimensional reduction approach to modulate the core ruminal microbiome associated with methane emissions via selective breeding.

The rumen is a complex microbial system of substantial importance in terms of greenhouse gas emissions and feed efficiency. This study proposes combining metagenomic and host genomic data for selective breeding of the cow hologenome toward reduced methane emissions. We analyzed nanopore long reads from the rumen metagenome of 437 Holstein cows from 14 commercial herds in 4 northern regions in Spain. After filtering, data were treated as compositional. The large complexity of the rumen microbiota was aggregated, through principal component analysis (PCA), into few principal components (PC) that were used as proxies of the core metagenome. The PCA allowed us to condense the huge and fuzzy taxonomical and functional information from the metagenome into a few PC. Bivariate animal models were applied using these PC and methane production as phenotypes. The variability condensed in these PC is controlled by the cow genome, with heritability estimates for the first PC of ~0.30 at all taxonomic levels, with a large probability (>83%) of the posterior distribution being >0.20 and with the 95% highest posterior density interval (95%HPD) not containing zero. Most genetic correlation estimates between PC1 and methane were large (≥0.70), with most of the posterior distribution (>82%) being >0.50 and with its 95%HPD not containing zero. Enteric methane production was positively associated with relative abundance of eukaryotes (protozoa and fungi) through the first component of the PCA at phylum, class, order, family, and genus. Nanopore long reads allowed the characterization of the core rumen metagenome using whole-metagenome sequencing, and the purposed aggregated variables could be used in animal breeding programs to reduce methane emissions in future generations.

Mycobacterium abscessus ventriculoperitoneal shunt infection and review of the literature.

A 30-year-old man with history of neonatal hydrocephalus requiring ventriculoperitoneal shunt placement presented with Mycobacterium abscessus shunt infection despite no shunt manipulation over 10 years prior to presentation. Cure was not achieved until complete removal of all CNS shunt foreign body was performed despite initial adequate antimicrobial therapy.

Fungal and ciliate protozoa are the main rumen microbes associated with methane emissions in dairy cattle.

BACKGROUND: Mitigating the effects of global warming has become the main challenge for humanity in recent decades. Livestock farming contributes to greenhouse gas emissions, with an important output of methane from enteric fermentation processes, mostly in ruminants. Because ruminal microbiota is directly involved in digestive fermentation processes and methane biosynthesis, understanding the ecological relationships between rumen microorganisms and their active metabolic pathways is essential for reducing emissions. This study analysed whole rumen metagenome using long reads and considering its compositional nature in order to disentangle the role of rumen microbes in methane emissions. RESULTS: The ß-diversity analyses suggested a subtle association between methane production and overall microbiota composition (0.01 < R2 < 0.02). Differential abundance analysis identified 36 genera and 279 KEGGs as significantly associated with methane production (Padj < 0.05). Those genera associated with high methane production were Eukaryota from Alveolata and Fungi clades, while Bacteria were associated with low methane emissions. The genus-level association network showed 2 clusters grouping Eukaryota and Bacteria, respectively. Regarding microbial gene functions, 41 KEGGs were found to be differentially abundant between low- and high-emission animals and were mainly involved in metabolic pathways. No KEGGs included in the methane metabolism pathway (ko00680) were detected as associated with high methane emissions. The KEGG network showed 3 clusters grouping KEGGs associated with high emissions, low emissions, and not differentially abundant in either. A deeper analysis of the differentially abundant KEGGs revealed that genes related with anaerobic respiration through nitrate degradation were more abundant in low-emission animals. CONCLUSIONS: Methane emissions are largely associated with the relative abundance of ciliates and fungi. The role of nitrate electron acceptors can be particularly important because this respiration mechanism directly competes with methanogenesis. Whole metagenome sequencing is necessary to jointly consider the relative abundance of Bacteria, Archaea, and Eukaryota in the statistical analyses. Nutritional and genetic strategies to reduce CH4 emissions should focus on reducing the relative abundance of Alveolata and Fungi in the rumen. This experiment has generated the largest ONT ruminal metagenomic dataset currently available.

Influence of age at first calving in a continuous calving season on productive, functional, and economic performance in a Blonde d'Aquitaine beef population.

The lifetime production of 7,655 cows with known age at first calving and a total of 27,118 parity records from 301 purebred Blonde d'Aquitaine herds were used to demonstrate the economic benefits of 2 yr of age at first calving. Ages at first calving ranged from 20 to 48 mo, and cows were divided into 5 calving groups, starting with early calving from age 20 to 27 mo up to late calving from age 40 to 48 mo. The information was gathered into 2 data sets, one for only primiparous cows and the second for all cows. The traits analyzed in this study were grouped as functional, linear type, and production traits. Functional traits were calving interval, calving ease, and number of calvings. Skeletal, muscle, and functional appraisal were included as linear type traits. The production traits studied were BW and weaning weight, carcass growth, and conformation of the offspring. The only significant traits found in primiparous cows were late age at first calving, which resulted in heavier BW calves, and early age at first calving, which resulted in calves with greater carcass conformation scores. Age at first calving was found to be significant only in its effect on BW and the number of calvings over a cow's lifetime, with lighter calves for early age at first calving. Heritability for age at first calving was 0.17. Genetic correlation of age at first calving with direct calving ease was positive (0.27) and that with maternal calving ease was negative (-0.39). Age at first calving showed a negative genetic correlation with lifetime number of calvings (-0.29) and a positive correlation with calving interval (0.14). Correlations with production and type traits were low, except for skeletal development (-0.29). Based on phenotypic and genetic analysis, there is a tendency for early-calving cows to produce a greater lifetime number of calves with less muscle but good carcass growth. Age at first calving affected the number of heifers in the herd, replacement rate, and number of animals slaughtered each year. Shortening the age at first calving from 3 to 2 yr led to a reduction of heifer feeding cost of US$21.24 (17.7€), a reduction of production cost of $26.52 (22.1€), and a profit increase of $25.80 (21.50€) per slaughtered animal per year over lifetime cow production.

Hydrogen Sulfide Improves Cardiomyocyte Function in a Cardiac Arrest Model.

BACKGROUND Cardioplegic arrest is a common procedure for many types of cardiac surgery, and different formulations have been proposed to enhance its cardio-protective effect. Hydrogen sulfide is an important signaling molecule that has cardio-protective properties. We therefore studied the cardio-protective effect of hydrogen sulfide in cardiac cell culture and its potential therapeutic use in combination with cardioplegia formulations. MATERIAL AND METHODS We added hydrogen sulfide donor GYY4137 to HL-1 cells to study its protective effect in nutrient starved conditions. In addition, we tested the potential use of GYY4137 when it is added into two different cardioplegia formulations: Cardi-Braun® solution and del Nido solution in an ex vivo Langendorff perfused rat hearts model. RESULTS We observed that eight-hour pre-treatment with GYY4137 significantly suppressed apoptosis in nutrient-starved HL-1 cells (28% less compared to untreated cells; p<0.05), maintained ATP content, and reduced protein synthesis. In ex vivo experiments, Cardi-Braun® and del Nido cardioplegia solutions supplemented with GYY4137 significantly reduced the pro-apoptotic protein caspase-3 content and preserved ATP content. Furthermore, GYY4137 supplemented cardioplegia solutions decreased the S-(5-adenosyl)-L-methionine/S-(adenosyl)-L-homocysteine ratio, reducing the oxidative stress in cardiac tissue. Finally, heart beating analysis revealed the preservation of the inter-beat interval and the heart rate in del Nido cardioplegia solution supplemented with GYY4137. CONCLUSIONS GYY4137 preconditioning preserved energetic state during starved conditions, attenuating the cardiomyocytes apoptosis in vitro. The addition of GYY4137 to cardioplegia solutions prevented apoptosis, ATP consumption, and oxidative stress in perfused rat hearts, restoring its electrophysiological status after cardiac arrest. These findings suggested that GYY4137 sulfide donor may improve the cardioplegia solution performance during cardiac surgery.

[Trends in coronary artery bypass surgery: changes in the profile of the surgical patient]. / Tendencias en cirugía coronaria: cambios en el perfil del paciente quirúrgico.

INTRODUCTION AND OBJECTIVES: In the last few years, the percentage of high-risk patients proceeding to coronary artery bypass surgery has increased. The most common risk factors are older age and the presence of comorbid complaints. We carried out a retrospective study to confirm this new risk profile and to evaluate its impact on surgical results. PATIENTS AND METHOD: We analyzed the changing risk profile of 1360 patients who underwent coronary artery bypass surgery in our hospital between 1993 and 2001, divided into three historical cohorts: 1993-1996, 1997-1999 and 2000-2001. The main factors associated with morbidity and mortality were analyzed by logistic regression analysis. The introduction of new operative techniques, such as off-pump surgery and arterial grafting, was also evaluated. RESULTS: The patients' risk profile worsened over time: patients were older, comorbid complaints were more common, and ventricular function was poorer. EuroSCORE figures reflected this trend: estimated mortality in the three historical cohorts was 2.0%, 4.0% and 4.2%, respectively (P<.001). However, risk-adjusted mortality, at 3.7%, 2.7% and 1.5%, respectively, decreased (P<.05), and combined overall morbidity and mortality remained stable, at 16.7%, 16.4% and 13.8%, respectively, (P<.39). There was a non-significant tendency for arterial grafting and off-pump surgery to reduce in-hospital morbidity and mortality. CONCLUSIONS: The risk profile of patients undergoing surgery has worsened as their mean age has increased and as comorbid complaints have become more prevalent. However, there has been no simultaneous increase in risk-adjusted mortality. The potential benefits of new surgical advances such as off-pump surgery and multiple arterial grafting must be corroborated by future studies.

[Right atrial myxoma in a patient with antiphospholipid antibodies]. / Mixoma auricular derecho en paciente con anticuerpos antifosfolipídicos positivos.

A 28 year-old asymptomatic male with previously detected elevation of antiphospholipid antibodies (aPL) had a right atrial mass diagnosed by transthoracic echocardiography. Although the morphological characteristics were highly suggestive of myxoma, due to the presence of aPL elevation it had to be differentiated from atrial thrombus. The patient underwent surgery and a 3 x 4 cm mass with its large base attached to the interatrial septum was removed. Histopathological examination confirmed that it was a myxoma. On follow-up, the patient was asymptomatic and antiphospholipid antibodies became undetectable.

[Advantages of off-pump coronary bypass surgery in high-risk patients]. / Ventajas de la revascularización miocárdica sin circulación extracorpórea en pacientes de riesgo.

INTRODUCTION AND OBJECTIVES: Myocardial revascularization without cardiopulmonary bypass has been shown to reduce operative morbi-mortality. We report our recent experience with this novel technique in order to evaluate its theoretical advantages in comparison with conventional surgery. PATIENTS AND METHODS: This retrospective analysis included 547 consecutive patients undergoing isolated myocardial revascularization from December 1997 through November 2000. One hundred twenty-one off-pump patients were compared to 426 undergoing cardiopulmonary bypass. Logistic regression analysis was performed to find predictors of mortality, transfusion, postoperative atrial fibrillation and length of hospital stay. RESULTS: Off-pump patients were at greater risk: they were older, with a lower ejection fraction and a higher prevalence of unstable angina, heart failure and associated comorbidity. Off-pump surgery reduced transfusions (1 1 vs 1,9 2 blood units; p < 0.0001) and postoperative hospital stays (8.9 5 vs 11,3 7 days; p < 0.001). The off-pump group showed a trend toward reduced morbidity but the technique did not decrease hospital mortality. Cardiopulmonary bypass was an independent predictor of blood transfusion and longer hospital stay. Short-term follow-up revealed no significant differences in recurring angina or patency rates. CONCLUSIONS: Off-pump coronary bypass surgery is a good option in high-risk patients because it reduces the incidence of perioperative transfusion and the length of hospitalization. Furthermore, it showed a trend toward reduced morbidity. Mortality was not significantly higher in spite of the higher risk of the patients. Long-term longitudinal follow-up is mandatory to assess the true effectiveness of this technique.

[Aortic valve replacement in patients over 70 years old: determinants of early death]. / Recambio valvular aórtico en pacientes mayores de 70 años: determinantes de mortalidad temprana.

INTRODUCTION AND OBJECTIVES: There is controversy regarding the risk factors associated with early death in geriatric patients undergoing aortic valve replacement. We analyzed the risks in these patients and established an accurate model for predicting in-hospital mortality. PATIENTS AND METHOD: Univariate and multivariate analyses were made of the risk factors associated with early death in a group of 129 patients older than 70 years who underwent aortic valve replacement (May 1994-June 2001). The variables obtained by multivariate logistic regression were combined to produce an equation for the prediction of early death. The equation was tested using a receiver operating characteristic curve. RESULTS: Univariate analysis identified four factors related to early death: NYHA III-IV (p < 0.0001), ejection fraction < 40% (p < 0.05), aortic regurgitation (p < 0.05) and high left ventricular mass index (p < 0.05). Multivariate analysis revealed three independent risk factors: NYHA III-IV (p < 0.01), aortic regurgitation (p < 0.05), and small body surface area (p < 0.05). A lower mortality was observed in patients with a larger body surface area (0% for > 1.90 m2, 20% for < 1.40 m2). The estimated mortality with the predictive model was 7.06%, which was similar to the observed mortality of 7.80% (area under the ROC curve 0.87) and better than estimates obtained with the EuroSCORE (6.5%; area under the ROC curve 0.56). CONCLUSIONS: Risk factors associated with early death after aortic valve replacement in geriatric patients include functional status, aortic regurgitation, and small body surface area. Our model based on these factors accurately predicted operative mortality in our patients. Gender, prosthesis size, and pump time were not identified as risk factors.

Triple cardiac rupture.

Left ventricular free wall rupture and acute ischaemic mitral regurgitation are nowadays rare, but still potentially lethal mechanical complications after acute myocardial infarction. We report a case of a sequential left ventricular free wall rupture, anterolateral papillary muscle disruption, secondary severe mitral regurgitation and subsequent posteromedial papillary muscle head rupture in a single patient during the same ischaemic episode after myocardial infarction, and their related successful surgical procedures and management until discharge. Prompt bedside diagnosis and emergent consecutive surgical procedures, as well as temporary left ventricular assistance, were crucial in the survival of this patient.

A comparison of electrospun polymers reveals poly(3-hydroxybutyrate) fiber as a superior scaffold for cardiac repair.

The development of biomaterials for myocardial tissue engineering requires a careful assessment of their performance with regards to functionality and biocompatibility, including the immune response. Poly(3-hydroxybutyrate) (PHB), poly(e-caprolactone) (PCL), silk, poly-lactic acid (PLA), and polyamide (PA) scaffolds were generated by electrospinning, and cell compatibility in vitro, and immune response and cardiac function in vitro and in vivo were compared with a noncrosslinked collagen membrane (Col) control material. Results showed that cell adhesion and growth of mesenchymal stem cells, cardiomyocytes, and cardiac fibroblasts in vitro was dependent on the polymer substrate, with PHB and PCL polymers permitting the greatest adhesion/growth of cells. Additionally, polymer substrates triggered unique expression profiles of anti- and pro-inflammatory cytokines in human peripheral blood mononuclear cells. Implantation of PCL, silk, PLA, and PA patches on the epicardial surface of healthy rats induced a classical foreign body reaction pattern, with encapsulation of polymer fibers and induction of the nonspecific immune response, whereas Col and PHB patches were progressively degraded. When implanted on infarcted rat heart, Col, PCL, and PHB reduced negative remodeling, but only PHB induced significant angiogenesis. Importantly, Col and PHB modified the inflammatory response to an M2 macrophage phenotype in cardiac tissue, indicating a more beneficial reparative process and remodeling. Collectively, these results identify PHB as a superior substrate for cardiac repair.

Cardiac differentiation is driven by NKX2.5 and GATA4 nuclear translocation in tissue-specific mesenchymal stem cells.

Myocardial infarction is a major public health problem that causes significant mortality despite recent advances in its prevention and treatment. Therefore, approaches based on adult stem cells represent a promising alternative to conventional therapies for this life-threatening condition. Mesenchymal stem cells (MSCs) are self-renewing pluripotent cells that have been isolated from multiple tissues and differentiate to various cell types. Here we have analyzed the capacity of MSCs from human bone marrow (BMSC), adipose tissue (ATSC), and dental pulp (DPSC) to differentiate to cells with a cardiac phenotype. Differentiation of MSCs was induced by long-term co-culture with neonatal rat cardiomyocytes (CMs). Shortly after the establishment of MSC-CM co-cultures, expression of connexin 43 and the cardiac-specific markers troponin I, beta-myosin heavy chain, atrial natriuretic peptide, and alpha-sarcomeric actinin was detected in BMSCs, ATSCs, and DPSCs. Expression of differentiation markers increased over time in the co-cultures, reaching the highest levels at 4 weeks. Translocation of the transcription factors NKX2.5 and GATA4 to the nucleus was observed in all three cultures of MSCs during the differentiation process; moreover, nuclear localization of NKX2.5 and GATA4 correlated with expression of alpha-sarcomeric actinin. These changes were accompanied by an increase in myofibril organization in the resulting CM-like cells as analyzed by electron microscopy. Thus, our results provide novel information regarding the differentiation of tissue-specific MSCs to cardiomyocytes and support the potential use of MSCs in cell-based cardiac therapies.

Purification process development for HER1 extracellular domain as a potential therapeutic vaccine.

HER1 is a tumor associated antigen emerging as an attractive target for cancer therapy. In the present study we demonstrated for first time that HER1 extracellular domain can be purified by a downstream process at pilot scale based on immunoaffinity chromatography from bioreactor supernatant of HEK 293 transfectomes. Filtered supernatant was applied to CNBr-activated Sepharose CL-4B with monoclonal antibody anti-human EGF immobilized, followed by three additional chromatographic polishing steps. HER1 extracellular domain was obtained with high purity (>95%), low DNA content, and biological activity.

[Instability from the distal radioulnar joint: reconstruction and evolution of dorsal triangular fibrocartilage complex ligament]. / Inestabilidad radiocubital distal: reconstrucción del ligamento dorsal del fibrocartílago triangular y su evolución.

Dorsal and palmar radio ulnar ligaments are part of the triangular fibrocartilage complex and are the principal stabilizer of this articulation. The distal radio ulnar joint provides not only pronation and supination to the forearm, but also it's basic for strength. The objective of this study is to evaluate the functional results of the dorsal radio ulnar joint ligament reconstruction in patient with distal radio ulnar instability. This is a prospective, longitudinal, experimental and auto-control assay. Functional preoperative data was analyzed, proper surgical technique and another findings. The patients were evaluated with a functional exploration, comparative dynamometry and DASH evaluation, they were also asked about their personal satisfaction. Between 1997 and 2004, this treatment was practiced in eleven patients (2 men and 9 women) with distal radio ulnar joint instability. With a mean age of 34.1 years. The dominant had was involved in 91% of patients. The mean complaint was pain for all patients, weakness in 9 patients and instability in 7. The mean follow up was 2 years 4 months, with a range from 6 months to 8 years. The patients refer totally satisfied, 8 of them, partially satisfied 2 and one patient insatisfied.

Virtual vascular endoscopy for acute aortic dissection.

Virtual endoscopy of the aorta is a new three-dimensional reconstruction method from multislice computed tomography or magnetic resonance that offers a virtual navigation through the aorta, and the possibility of having a new preoperative endoluminal vision. We present a case of subacute aortic dissection with a preoperative virtual endoscopy of the aorta.

Biatrial radiofrequency ablation for atrial fibrillation: epicardial and endocardial surgical approach.

Radiofrequency energy applied by means of surgical probes permits the ablation of atrial fibrillation (AF). This study presents our initial experience on 55 consecutive cardiac patients with permanent AF with radiofrequency ablation through biatrial epicardial and endocardial surgical approach. At discharge, 8.1% of the patients had persistent AF-atrial flutter. Hospital incidence of arrhythmias were, 9% of paroxysmal atrial fibrillation, 10.9% of atrial flutter, and 34.5% of persistent atrial fibrillation. After a mean follow-up of 7 months, 83.6% patients have recovered sinus rhythm, and echocardiographic biatrial contraction was re-established in 73%. Biatrial radiofrequency ablation applied from the endocardium and the epicardium has achieved satisfactory results, without increasing the surgical risk.