Application of a digital PCR method for WT1 to myeloid neoplasms in CR and deep ELN WT1 molecular response (< 10 copies).

Bone marrow WT1 mRNA levels assessed by the ELN method are useful to establish prognostic correlations in myeloid malignancies treated with chemotherapy or hematopoietic stem cell transplantation (HCT). Those patients with WT1 levels below ten copies have a good outcome. However, some of these patients relapse. To further characterize this group of cases, we applied a new and sensitive digital (ddPCR) WT1 method. A consecutive series of 49 patients with treated myeloid malignancies and with an ELN WT1 quantitation of < 10 copies were included in the study. All cases (47 AML and 2 MDS) have received intensive chemotherapy or HCT. One to four micrograms of total RNA were retrotranscribed to obtain ≥ 10,000 ABL1 copies using the ELN protocol. Only those cases with a good quality cDNA were used in the ddPCR WT1 test. The ddPCR Gene Expression WT1 Assay of Bio-Rad© was used to perform the PCR amplification, and the microdroplets were quantified in the Bio-Rad's QX200 droplet reader. Eighteen patients showed a negative WT1 ddPCR assay (0 copies/µl), whereas 31 cases were positive (results ranged from 1 to 15.2 copies/µl). Survival analysis showed statistically significant differences in terms of OS between both groups, 83 ± 8% vs. 46 ± 9% (p = 0.024). A statistically significant correlation was also found between ddPCRWT1 results and CD123+ cell number detected by flow cytometry (p = 0.024). Larger series of patients tested with the current ddPCRWT1 method will solve whether it could be used to stratify patients with myeloid malignancies achieving deep WT1 molecular response (< 10 copies).

Phase Diagram of Fatty Acid Langmuir Monolayers from Rheological Measurements.

Langmuir monolayers of fatty acids and alcohols are two-dimensional systems with a rich equilibrium phase diagram. We have explored the temperature and surface-pressure-dependent shear response of monolayers formed by fatty acids of different chain lengths and a fatty alcohol. This has been accomplished with an interfacial shear rheometer utilizing magnetic tweezers and equipped with a refined temperature control and acquisition system. Our rheological results have allowed us to draw a phase diagram from the viscoelastic properties of these 2-D systems and show new phenomena that strongly depend on temperature: the existence of a maximum in viscosity at the L2' phase, the behavior of the elastic modulus to the storage modulus ratio at the L2 phase, and the increase or decrease in viscosity at the L2-LS phase transition. In addition, we unambiguously show that the LS phase displays a counterintuitive behavior in which the loss modulus increases with temperature. We demonstrate, through isothermal surface pressure sweeps and isobaric temperature sweeps, that the exponential dependence of the loss modulus on temperature at the LS phase appears for all hydrophobic tail lengths studied and for both acid and alcohol head groups.

Magnetic microwire probes for the magnetic rod interfacial stress rheometer.

The magnetic needle interfacial shear rheometer is a valuable tool for the study of the mechanical properties of thin fluid films or monolayers. However, it is difficult to differentiate the interfacial and subphase contributions to the drag on the needle. In principle, the problem can be addressed by decreasing the needle diameter, which decreases the bulk contribution while the interfacial contribution remains essentially the same. Here we show the results obtained when using a new type of needle, that of magnetic microwires with diameter approximately 10 times thinner than for commercial needles. We show that the lower inertia of the microwires calls for a new calibration procedure. We propose such a new calibration procedure based on the flow field solution around the needle introduced in refs 1 and 2. By measuring thin silicone oil films with well-controlled interfacial viscosities as well as eicosanol (C20) and pentadecanoic acid (PDA, C15) Langmuir monolayers, we show that the new calibration method works well for standard needles as well as for the microwire probes. Moreover, we show that the analysis of the force terms contributing to the force on the needle helps to ascertain whether the measurements obtained are reliable for given surface shear viscosity values. We also show that the microwire probes have at least a 10-fold-lower resolution limit, allowing one to measure interfacial viscosities as low as 10(-7) N·m/s.

[Can cough peak flow be used to assess cough capacity in severe obstructive respiratory diseases?] / ¿Se puede utilizar la determinación del pico flujo de tos para valorar la capacidad tusígena en enfermedades respiratorias obstructivas graves?

INTRODUCTION: Cough capacity is assessed by measuring cough peak flow (CPF). However, this assessment could be altered by obstructive airway diseases. The aim was to assess measurement of cough capacity by CPF in patients with chronic obstructive pulmonary disease (COPD), as well as correlations with pulmonary, respiratory muscle, and oropharyngeal function. METHODS: Patients with COPD, and with neuromuscular disease, were selected as well as healthy subjects who had undergone respiratory muscle strength measurement in a healthcare setting. From this population, respiratory function values and lung and oropharyngeal muscle function were analysed. A subgroup of COPD patients underwent a videofluoroscopic swallow study. RESULTS: Three hundred and seven subjects were included (59.3% COPD, 38.4% neuromuscular diseases, and 2.3% healthy). CPF was found to be statistically significantly decreased in the COPD group compared to both the healthy and neuromuscular disease groups. Of the COPD patients, 70% had a pathological decrease in CPF. There was only a direct correlation between CPF with the degree of bronchial obstruction and expiratory muscle strength. No alteration of inspiratory or oropharyngeal muscle function was found. CONCLUSIONS: The use of CPF in COPD patients does not reflect cough capacity as it is influenced by the degree of bronchial obstruction. Therefore, new diagnostic tests to measure cough capacity should be considered, especially in patients with coexisting neuromuscular diseases and severe bronchial obstructive disease.

Phase behavior of dense colloidal binary monolayers.

In this work, we study how structures develop on 2D dense binary colloidal monolayers as a function of the relative concentration of small/large particles. Translational and orientational distribution functions have been used to monitor the continuous phase transition through a detailed characterization of the global and local order. We have observed how a gradual enhancement in the number of particles of different sizes leads to a continuous vitrification process and how homogeneous binary glasses form in equimolar mixtures. Also, we have performed a simple calculation that relates the structures found to the pair dipolar potential, allowing the forecast of local structures in other arbitrary binary mixtures. Finally, we have corroborated the goodness of the binary systems as a glass-forming model by comparing the established scenario with the structural features found in partially aggregated monolayers.

Aggregation and disaggregation dynamics of sedimented and charged superparamagnetic micro-particles in water suspension.

This article presents results on the aggregation and disaggregation kinetics on a 1 µm diameter charged superparamagnetic particles dispersed in water under a constant uniaxial magnetic field in experiments with salt (KCl) added to the suspension in order to observe the behaviour of the system when the electrical properties of the particles have been screened. These particles have an electric charge and are confined between two separated 100 µm thick quartz windows, and sediment near the charged bottom wall. The electrostatic interactions that take place in this experimental setup may affect the micro-structure and colloidal stability of the suspension and thus, the dynamics of aggregation and disaggregation.

Rheology of poly(methyl methacrylate) Langmuir monolayers: percolation transition to a soft glasslike system.

An experimental study of the equilibrium properties and of the surface rheology of Langmuir monolayers of poly(methyl methacrylate) (PMMA) at the air/water interface has been carried out as a function of polymer concentration (Γ) and molecular weight (M(w)). Dilational and shear complex elasticity moduli covering a frequency range from 10(-3) to 0.2 Hz have been discussed. It was found that the air∕water interface behaves as a poor solvent for PMMA monolayers, thus suggesting that the polymer coils take collapsed soft-disks (pancakes) shape at the interface. The equilibrium and dynamic results suggest a fluid-to-soft-glass transition as the polymer concentration increases above a critical packing fraction at constant temperature. This two-dimensional transition is in agreement with results previously discussed for the dilational rheology of poly(4-hydroxystyrene) [F. Monroy, F. Ortega, R. G. Rubio, H. Ritacco, and D. Langevin, J. Chem. Phys. 95, 056103 (2005)]. Furthermore, the Γ-dependence of the relaxation dynamics of the monolayers suggests that the gel state may be considered as a fragile soft glass.

Gentamycin inhibits the growth of Malassezia pachydermatis in culture.

BACKGROUND: Malassezia pachydermatis is a yeast of importance in both veterinary and human medicine. AIMS: To know if M. pachydermatis grow on mycological media with high concentrations of gentamycin. METHODS: Twenty M. pachydermatis strains were streaked on Sabouraud Dextrose Agar plates with different concentrations of gentamycin. RESULTS: All isolates were inhibited when high concentrations of gentamycin were added. CONCLUSIONS: The use of plates with high concentrations of gentamycin can lead to some important misdiagnoses: firstly, false-negative cultures, and secondly, an erroneous classification of M. pachydermatis as a lipid-dependent species. Moreover, all of this could be useful in two therapeutic fields: i) in animals, topical gentamycin could be an efficacious treatment for a disease such as external otitis in dogs; ii) in humans, we hypothesize that gentamycin could be regarded as a possible therapy ("antibiotic-lock") for catheter-associated Malassezia spp. infections.

[The peritoneal kinetic study performed with hypertonic glucose permits better evaluation of UF capacity and determination of sieving of sodium]. / Las cinéticas con glucosa hipertónica permiten identificar mejor el fallo de ultrafiltración. ¿Qué aporta el cribado de sodio?

INTRODUCTION: The use of solutions containing hypertonic glucose (3.86%/4.25%) has been postulated as the method of choice for study the peritoneal function, and permits a better evaluation of the ultrafiltration (UF) capacity. OBJECTIVE: The aim of our study was to analyze the UF capacity and its relation with the peritoneal permeability and sieving of sodium, performing the peritoneal kinetic study with hypertonic glucose solutions. PATIENTS AND METHODS: We performed 184 peritoneal kinetic studies with hypertonic glucose solutions in stable patients on peritoneal dialysis (PD), with a mean time on PD of 16 +/- 22 months. We measured the mass transfer coefficient of creatinine (CrMTC), dialysate to plasma ratio of creatinine (D/PCr), UF capacity and sieving of sodium at 60 minutes (difNa60). RESULTS: The mean values were: CrMTC: 9.1 +/- 4.5 ml/min, D/PCr: 0.71 +/- 0.09, UF 759 +/- 233 ml/4 h and difNa60: 4.7 +/- 2.3. The best multivariate model that predicts the UF capacity included: difNa60, CrMTC, age and time on PD (r = 0.57; p > 0.0001). In patients with UF lower than 600 ml/4 h (Percentil 25) the correlation between UF and CrMTC was lost, but remains the correlation with difNa60 (r = 0.48). The patients with previous peritonitis (n = 38) showed no differences in UF, CrMTC or D/Pcr, but the had lower difNa60 (3.7 +/- 2.8 vs. 4.9 +/- 2.1; p = 0.002) than the remaining patients. CONCLUSIONS: The peritoneal kinetic study performed with hypertonic glucose allows to standardize the UF capacity and by determination of sieving of sodium, the early detection of water transport alterations, before the UF capacity and small solutes permeability alteration develops.

Nafcillin degradation by heterogeneous electro-Fenton process using Fe, Cu and Fe/Cu nanoparticles.

Heterogeneous electro-Fenton (HEF) is as an alternative to the conventional electro-Fenton (EF) process. HEF uses a solid phase catalyst, whereas EF employs a solubilized one. This implies that in HEF, material can be recovered through a simple separation process such as filtration or magnetic separation in HEF. HEF also has the advantage of not requires a previous pH adjustment, which facilitates working in a higher pH range. In this work, Fe, Cu and Fe/Cu bimetallic nanoparticles (Fe/Cu NPs) were synthesized, characterized and used for the degradation of Nafcillin (NAF). The effect of the adsorption and the anodic oxidation (AO-H2O2) process was tested to assess their influence on HEF. NAF adsorption did not exceed 24% of antibiotic removal and the AO-H2O2 process eliminated the total NAF after 240 min of electrolysis. Through the HEF process, the antibiotic was completely removed using Fe/Cu NPs after 7.0 min of electrolysis, while these NPs, mineralization reached 41% after 240 min. In this case, NAF degradation occurs mainly due to the generation of hydroxyl radicals in the BDD electrode, and the Fenton reaction with Fe and Cu NPs. The main organic intermediates produced during the degradation of NAF by HEF were identified allowing the proposal of degradation pathway. Finally, the antibiotic was also completely eliminated from a wastewater from slaughterhouse after 15 min of treatment by HEF and using Fe/Cu bimetallic NPs.

Effects of hTERT on genomic instability caused by either metal or radiation or combined exposure.

Genomic instability is considered to be an important component in carcinogenesis. It can be caused by low-dose exposure to agents, which appear to act through induction of stress-response pathways related to oxidative stress. These agents have been studied mostly in the radiation field but evidence is accumulating that chemicals, especially heavy metals such as Cr (VI), can also act in the same manner. Previous work showed that metal ions could initiate long-term genomic instability in human primary fibroblasts and this phenomenon was regulated by telomerase. The aim of this study was to examine the difference in clonogenic survival and cytogenetic damage after exposure to Cr (VI) and radiation both singly and in combination in normal human fibroblasts (hTERT- cells) and engineered human fibroblasts, infected with a retrovirus carrying a cDNA encoding hTERT, which rendered these cells telomerase positive and replicatively immortal (hTERT+ cells). Cr (VI) induced genomic instability in hTERT- cells but not in hTERT+ cells, whereas radiation induced genomic instability in hTERT+ cells and to a lesser extent in hTERT- cells. Combined exposure caused genomic instability in both types of cells. However, this genomic instability was more pronounced in hTERT- cells after radiation followed by Cr (VI) and more pronounced in hTERT+ cells after Cr (VI) followed by radiation. Moreover, the biological effects provoked by combined exposure of Cr (VI) and radiation also led to a synergistic action in both types of cells, compared to either Cr (VI) treatment only or radiation exposure only. This study suggests that telomerase can prevent genomic instability caused by Cr (VI), but not by radiation. Furthermore, genomic instability may be prevented by telomerase when cells are exposed to radiation and then Cr (VI) but not after exposure to Cr (VI) and then radiation.

A Spanish Society joint SECO and SEEDO approach to the Post-operative management of the patients undergoing surgery for obesity.

PURPOSE: Bariatric surgery is the method of choice for the management or treatment of obesity. Bariatric surgery brings about several physiological changes in the body and is associated with set of complications. The aim of this study is to provide guidelines on post bariatric surgery management based on consensus by the Spanish society for Obesity Surgery (Sociedad Española de Cirugía de la Obesidad) (SECO) and the Spanish Society for the Study of Obesity (Sociedad Española para el Estudio de la Obesidad) (SEEDO). METHOD: The boards proposed seven experts from each society. The experts provided the evidence and a grade of recommendation on the selected topics based on systematic reviews/meta-analysis. A list of clinical practical recommendations levels of evidence and grades of these recommendations was derived from the consensus statements from the members of these societies. RESULTS: Seventeen topics related to post-operative management were reviewed after bariatric surgery. The experts came with 47 recommendations and statements. The mean number of persons voting at each statement was 54 (range 36-76). CONCLUSION: In this consensus, we have designed a set of guidelines to be followed while managing patients after bariatric surgery. Expertise and knowledge of the clinicians are required to convey suitable considerations to the post-bariatric patients. There should also be extensive follow-up plans for the bariatric surgery patients.

Green tea reduces LDL oxidability and improves vascular function.

OBJECTIVE: Several different epidemiological studies have examined the association between the consumption of tea and coronary heart disease. Some, though not all, support the view that tea or flavonoids reduce the risk of cardiovascular heart disease. The aim of this study was to determine the short-to medium-term effect of a green tea extract on vascular function and lipid peroxidation as compared with placebo. METHODS: The study was undertaken with 14 healthy women, none of whom were receiving any medical treatment. Measurements were made of antibodies and immune complexes by ELISA, endothelial dependent vascular function by Doppler ultrasound, and the concentration of oxidized LDL by TBARS. RESULTS: The mean diameter of the brachial artery following the post-compression hyperaemia phase rose significantly (p < 0.0001) after treatment with green tea extract. Flow-mediated brachial artery vasodilation ranged from 5.68% for the placebo phase to 11.98% after the green tea extract (p = 0.02). The consumption of green tea extract was associated with a significant 37.4% reduction in the concentration of oxidized LDL (TBARS) (p = 0.017). The levels of anti-oxidized LDL IgM antibodies fell significantly after treatment (p = 0.002). CONCLUSION: This study found that consumption of green tea extract by women for five weeks produced modifications in vascular function and an important decrease in serum oxidizability.

Design and validation of a scale to assess preferences of type 2 diabetic patients towards different nutritional supplements.

OBJECTIVES: To design and validate a scale to evaluate preferences of type 2 diabetic patients towards nutritional supplements (Madrid scale) and to discover those taste attributes that are more discriminating. CONTEXT: ambulatory patients with type 2 diabetes mellitus. MATERIALS AND METHODS: 18 controls and 106 type 2 diabetic patients received 2 of the 7 stimuli studied (6 nutritional supplements and a differential salty stimulus) and then completed both scales and a criterion question. Two weeks later, 30 diabetic patients received a retest. The psychometric properties of the Madrid scale were studied and the relative importance of each stimuli attribute was assessed. RESULTS: Feasibility: The Madrid scale consists of 8 questions and is completed in less than five minutes; Dimensionality: A single dimension which explains 45.1% of the variance. Reliability: Cronbach's , 0.806; intraclass correlation coefficient, 0.835 (95% confidence interval: 0.653-0.922). Concurrent validity: Correlation indexes of the corrected total score with the criterion question and the Modified Wine-Tasting Scale, 0.731 (p < 0.0005) and 0.774 (p < 0.0005), respectively. The scale discriminated between subjects younger and older than 75 years and between supplements and the differential stimulus. Preferences: Glucerna SR chocolate, Glucerna SR strawberry, Glucerna SR vanilla, Diasip vanilla, Clinutren vanilla and Resource diabet vanilla. CONCLUSION: The Madrid scale has adequate psychometric properties for its use in research and daily clinical practice.

Relevance of the pyramidal syndrome in amyotrophic lateral sclerosis. / Relevancia del síndrome piramidal en la esclerosis lateral amiotrófica.

INTRODUCTION: Pyramidal signs (hyperreflexia, spasticity, Babinski sign) are essential for the diagnosis of amyotrophic lateral sclerosis (ALS). However, these signs are not always present at onset and may vary over time, besides which their role in disease evolution is controversial. Our goal was to describe which pyramidal signs were present and how they evolved in a cohort of patients with ALS, as well as their role in prognosis. METHODS: Retrospective analysis of prospectively collected patients diagnosed with ALS in our centre from 1990 to 2015. RESULTS: Of a total of 130 patients with ALS, 34 (26.1%) patients showed no pyramidal signs at the first visit while 15 (11.5%) had a complete pyramidal syndrome. Of those patients without initial pyramidal signs, mean time of appearance of the first signs was 4.5 months. Babinski sign was positive in 64 (49.2%) patients, hyperreflexia in 90 (69.2%) and 22 (16.9%) patients had spasticity. Pyramidal signs tended to remain unchanged over time, although they seem to appear at later stages or even disappear with time in some patients. We found no association between survival and the presence of changes to pyramidal signs, although decreased spasticity was associated with greater clinical deterioration (ALSFR scale) (P<.001). CONCLUSION: A quarter of patients with ALS initially showed no pyramidal signs and in some cases they even disappear over time. These data support the need for tools that assess the pyramidal tract.