An improved protocol for bacteria identification by MALDI-TOF MS directly from positive blood cultures.

FASTinov® developed a rapid antimicrobial susceptibility test that includes the purification of a bacterial suspension directly from positive blood cultures (BC). In order to streamline laboratory workflow, the use of the bacterial suspension obtained through FASTinov® sample prep was tested for identification (ID) by matrix absorption laser deionization-time of flight mass spectrometry (MALDI-TOF MS) (Bruker) in 364 positive BC, and its accuracy assessed comparing with the MALDI-TOF MS ID of the next-day subcultured colonies. FASTinov sample prep was highly reliable for rapid ID directly from BC with proportion of agreement of 94.9% for Gram-positive and 96.3% for Gram-negative bacteria.

Monoclonal antibody against Nile tilapia (Oreochromis niloticus) IgM heavy chain: A valuable tool for detection and quantification of IgM and IgM+ cells.

Nile tilapia (Oreochromis niloticus) is a freshwater fish, which is extensively cultivated worldwide and constitutes one of the model species for the study of fish immunology. Monoclonal antibodies are very advantageous molecular tools for studying teleost immune system. Specifically, monoclonal antibodies that react with immunoglobulins are used successfully in the study of the humoral immune response of several fish species. In the present study, we produced and characterized a monoclonal antibody against tilapia IgM heavy chain using a peptide-based strategy. The peptide sequence was selected from the surface-exposed region between CH3-CH4 domains. The specificity of the polyclonal serum and the hybridoma culture supernatant obtained by immunization with the peptide conjugated to keyhole limpet hemocyanin were evaluated by western blotting, both showing reactivity against tilapia serum IgM. The purified mAb was able to recognize secreted IgM by western blotting and ELISA and membrane IgM by flow cytometry. We also demonstrated that the antibody doesn't cross-react with a recombinant IgT fragment. This tool allowed us to study for the first time the stimulation of mucosal immunity after Pituitary Adenylate Cyclase Activating Polypeptide administration. Overall, the results demonstrated the utility of this mAb to characterize humoral immune response in O. niloticus.

d-Amino acid substitutions and dimerization increase the biological activity and stability of an IL-15 antagonist peptide.

Interleukin (IL)-15 plays an important role in several inflammatory diseases. We have previously identified an IL-15 antagonist called P8 peptide, which binds specifically to IL-15 receptor alpha subunit. However, the P8 peptide rapidly degraded by proteases, limiting its therapeutic application. Thus, we replaced each P8 peptide l-amino acid by its corresponding d-isomers. First, we determined the biological activity of the resulting peptides in a proliferation assay by using CTLL-2 cells. The substitution of l-Ala by d-Ala ([A4a]P8 peptide) increased the inhibitory effect of the P8 peptide in CTLL-2 cells in five-fold. In addition to that, the [A4a]P8 peptide dimer showed the most inhibitory effect. To protect the [A4a]P8 peptide and its dimer against exopeptidase activity, we acetylated the N-terminal of these peptides. At least a three-fold reduction in antagonist activity of acetylated peptides was exhibited. However, the substitution of the N-terminal l-Lys residue of [A4a]P8 peptide and its dimer by d-Lys ([K1k;A4a]P8 peptide) did not affect the antagonist effect of the aforementioned peptides. The [K1k;A4a]P8 peptide dimer was stable to the degradation of trypsin, chymotrypsin, and pepsin up until 48 min. Also, the safety and immunogenicity studies in healthy BALB/c mice demonstrated that the administration of this peptide did not affect the clinical parameters of the animals nor generated antipeptide antibodies. Our findings reveal that two distinct d-amino acid substitutions and dimerization increase the biological activity and stability of P8 peptide. The resulting peptide constitutes a novel IL-15 antagonist with potential applicability in inflammatory diseases.

Low-Cost Ultrasonic Range Improvements for an Assistive Device.

To achieve optimal mobility, visually impaired people have to deal with obstacle detection and avoidance challenges. Aside from the broadly adopted white cane, electronic aids have been developed. However, available electronic devices are not extensively used due to their complexity and price. As an effort to improve the existing ones, this work presents the design of a low-cost aid for blind people. A standard low-cost HC-SRF04 ultrasonic range is modified by adding phase modulation in the ultrasonic pulses, allowing it to detect the origin of emission, thus discriminating if the echo pulses come from the same device and avoiding false echoes due to interference from other sources. This improves accuracy and security in areas where different ultrasonic sensors are working simultaneously. The final device, based on users and trainers feedback for the design, works with the user's own mobile phone, easing utilization and lowering manufacturing costs. The device was tested with a set of twenty blind persons carrying out a travel experiment and satisfaction survey. The main results showed a change in total involuntary contacts with unknown obstacles and high user satisfaction. Hence, we conclude that the device can fill a gap in the mobility aids and reduce feelings of insecurity amongst the blind.

A Non-Intrusive Approach for Indoor Occupancy Detection in Smart Environments.

Smart Environments try to adapt their conditions focusing on the detection, localisation, and identification of people to improve their comfort. It is common to use different sensors, actuators, and analytic techniques in this kind of environments to process data from the surroundings and actuate accordingly. In this research, a solution to improve the user's experience in Smart Environments based on information obtained from indoor areas, following a non-intrusive approach, is proposed. We used Machine Learning techniques to determine occupants and estimate the number of persons in a specific indoor space. The solution proposed was tested in a real scenario using a prototype system, integrated by nodes and sensors, specifically designed and developed to gather the environmental data of interest. The results obtained demonstrate that with the developed system it is possible to obtain, process, and store environmental information. Additionally, the analysis performed over the gathered data using Machine Learning and pattern recognition mechanisms shows that it is possible to determine the occupancy of indoor environments.

Noisy nonlinear dynamics of vesicles in flow.

We present a model for the dynamics of fluid vesicles in linear flow which consistently includes thermal fluctuations and nonlinear coupling between different modes. At the transition between tank treading and tumbling, we predict a trembling motion which is at odds with the known deterministic motions and for which thermal noise is strongly amplified. In particular, highly asymmetric shapes are observed even though the deterministic flow only allows for axisymmetric ones. Our results explain quantitatively recent experimental observations [Levant and Steinberg, Phys. Rev. Lett. 109, 268103 (2012)].

Monoclonal antibody generated against Nile tilapia (Oreochromis niloticus) IgT heavy chain using a peptide-based strategy.

Teleost IgT/Z plays a principal role in the defense mechanisms against infectious agents in the mucosal compartments and in systemic immunity. Previously, Nile tilapia (Oreochromis niloticus) IgT was discovered and characterized at transcription level. In this work, we generated a monoclonal antibody (mAb) that specifically recognized the Nile tilapia IgT. BALB/c mice were immunized with three synthetic peptides conjugated to KLH. The sequences of these peptides derived from the constant region of the Nile tilapia IgT heavy chain. ELISA and Western blotting confirmed the specificity of the polyclonal sera and the culture supernatant from a positive hybridoma clone. We observed immunoreactivity against a recombinant IgT fragment and native IgT in skin mucus. The anti-IgT mAb did not cross-react with purified tilapia IgM. Direct ELISA analysis allowed the quantification of skin mucus IgM and IgT concentrations. Flow cytometry analysis revealed differences in the percentage of IgT+ B cell populations between juveniles and adults in peripheral blood, head kidney and spleen lymphocytes and among the tissues analyzed. For further validation of the anti-IgT mAb utility, a recombinant vaccine candidate against sea lice (TT-P0 Ls) was injected into juvenile tilapia. Direct ELISA results revealed a differential secretion of skin mucus IgT and IgM after immunostimulation. In addition, the percentages of IgT+ B cells were determined at 7 days after booster and ex-vivo stimulation by flow cytometry. This mAb constitutes an important immunological tool to study the biological function and structural characteristics of tilapia IgT.

Thermodynamics of genuine nonequilibrium states under feedback control.

For genuine nonequilibrium states that even at fixed external control parameter exhibit dissipation, we extend the Hatano-Sasa equality to processes with feedback control. The resulting bound on the maximal extractable work is substantially sharper than what would follow from applying the Sagawa-Ueda equality to transitions involving such states. For repeated measurements at short enough intervals, the power thus extracted can even exceed the average cost of driving as demonstrated explicitly with a simple, analytically solvable example.

Antioxidants from barley husks impregnated in films of low-density polyethylene and their effect over lipid deterioration of frozen cod (Gadus morhua).

BACKGROUND: The changes in quality of cod fillets packaged in films with and without antioxidants during 12 months of frozen storage at - 20 °C were investigated in the present study. The following parameters were determined in order to study lipid hydrolysis and primary and secondary lipid oxidation in the samples during frozen storage: peroxide value, conjugated dienes, conjugated triene hydroperoxides, free fatty acids, totox value, thiobarbituric acid-reactive substances and p-anisidine value. RESULTS: Films containing antioxidants isolated from barley husks were effective in slowing down lipid hydrolysis and primary and secondary lipid oxidation processes. Secondary lipid oxidation reached maximum values in the 12th month of storage in control samples and samples packaged with antioxidant-containing film. Maximum lipid hydrolysis and lipid oxidation values for control cod samples were significantly higher than the maximum values found in samples packaged with antioxidant-containing film. CONCLUSION: The results confirm the efficacy of natural antioxidants derived from barley husks in slowing down lipid hydrolysis and increasing the oxidative stability of cod flesh. They also demonstrate the potential usefulness of natural antioxidants extracted from barley husks in the development of active packaging films for food preservation.

Validation of an eBAT as a mobility aid for blind people.

One of the challenges faced by blind persons to achieve optimal mobility is the detection and avoidance of obstacles located in their travel path. Besides the widely used white cane, alternative or complementary devices have been developed, such as electronic aids that provide feedback about the environment. However, the devices available have been unable to provide an optimal solution with widespread acceptance, motivating the present work. The eBAT (electronic Buzzer for Autonomous Travel) is designed to offer optimal protection and employs the user's own mobile phone for easier use and reduced manufacturing costs. For this work, a group of 25 blind individuals was used to validate the eBAT based on the single-subject with reversal method (ABA study). The results show a significant decrease in the number of involuntary contacts in an unknown travel path between the first phase of the study, which did not involve the eBAT, and the second, where it was used. When the device was again removed in the third phase, the number of contacts rose. We may therefore conclude that the eBAT fills an important gap in mobility aids for blind people, yielding a clear benefit by reducing the participants' feeling of insecurity.

Improving agricultural knowledge management: The AgTrials experience.

Background: Opportunities to use data and information to address challenges in international agricultural research and development are expanding rapidly. The use of agricultural trial and evaluation data has enormous potential to improve crops and management practices. However, for a number of reasons, this potential has yet to be realized. This paper reports on the experience of the AgTrials initiative, an effort to build an online database of agricultural trials applying principles of interoperability and open access. Methods: Our analysis evaluates what worked and what did not work in the development of the AgTrials information resource. We analyzed data on our users and their interaction with the platform. We also surveyed our users to gauge their perceptions of the utility of the online database. Results: The study revealed barriers to participation and impediments to interaction, opportunities for improving agricultural knowledge management and a large potential for the use of trial and evaluation data.  Conclusions: Technical and logistical mechanisms for developing interoperable online databases are well advanced.  More effort will be needed to advance organizational and institutional work for these types of databases to realize their potential.

Incubation of boar spermatozoa in viscous media by addition of methylcellulose improves sperm quality and penetration rates during in vitro fertilization.

This work was designed to study whether viscous media can improve the in vitro sperm functionality in pigs by using methylcellulose as a thickener. Viscosity of porcine oviductal fluid (POF) was compared with culture medium (Tyrode's) supplemented with methylcellulose (MET 0, 0.5 and 1% w/v). Spermatozoa were incubated in the different media (0, 1 and 2 h) and sperm motion parameters, lipid membrane disorder, plasma membrane integrity and reactive oxygen species (ROS) formation were assessed. Fertilization results were assessed i) preincubating spermatozoa in the viscous media followed by gamete coculture in a non-viscous medium; and ii) gamete coculture in the viscous media. Viscosity of POF from early luteal phase was higher than late follicular phase. Medium without methylcellulose presented constant viscosity with increased shear rate, while viscosity of the POF and media with methylcellulose was reduced by increased shear rates. Methylcellulose improved sperm linearity, straightness and the proportion of fast-linear spermatozoa. Moreover, methylcellulose increased the rate of viable spermatozoa with intact acrosome and low lipid disorder, reducing the ROS generation. Preincubation in viscous media increased the penetration rate and the mean number of spermatozoa bound to the zona pellucida (both with 0.5 and 1% MET) and reduced monospermy with 1% MET. On the other hand fertilization in the viscous media reduced penetration rate and increased monospermy. The efficiency of the IVF system was not improved with the use of viscous media. The results show the relevance of increasing viscosity thus making the in vitro media more comparable to physiological conditions.

Fluid vesicles in flow.

We review the dynamical behavior of giant fluid vesicles in various types of external hydrodynamic flow. The interplay between stresses arising from membrane elasticity, hydrodynamic flows, and the ever present thermal fluctuations leads to a rich phenomenology. In linear flows with both rotational and elongational components, the properties of the tank-treading and tumbling motions are now well described by theoretical and numerical models. At the transition between these two regimes, strong shape deformations and amplification of thermal fluctuations generate a new regime called trembling. In this regime, the vesicle orientation oscillates quasi-periodically around the flow direction while asymmetric deformations occur. For strong enough flows, small-wavelength deformations like wrinkles are observed, similar to what happens in a suddenly reversed elongational flow. In steady elongational flow, vesicles with large excess areas deform into dumbbells at large flow rates and pearling occurs for even stronger flows. In capillary flows with parabolic flow profile, single vesicles migrate towards the center of the channel, where they adopt symmetric shapes, for two reasons. First, walls exert a hydrodynamic lift force which pushes them away. Second, shear stresses are minimal at the tip of the flow. However, symmetry is broken for vesicles with large excess areas, which flow off-center and deform asymmetrically. In suspensions, hydrodynamic interactions between vesicles add up to these two effects, making it challenging to deduce rheological properties from the dynamics of individual vesicles. Further investigations of vesicles and similar objects and their suspensions in steady or time-dependent flow will shed light on phenomena such as blood flow.

Effect of thermal noise on vesicles and capsules in shear flow.

We add thermal noise consistently to reduced models of undeformable vesicles and capsules in shear flow and derive analytically the corresponding stochastic equations of motion. We calculate the steady-state probability distribution function and construct the corresponding phase diagrams for the different dynamical regimes. For fluid vesicles, we predict that at small shear rates thermal fluctuations induce a tumbling motion for any viscosity contrast. For elastic capsules, due to thermal mixing, an intermittent regime appears in regions where deterministic models predict only pure tank treading or tumbling.

Dieta cetogénica en el tratamiento de la epilepsia refractaria en niños / Ketogenic diet in the treatment of the refractory epilepsy in children

Se realizó un estudio descriptivo con el objetivo de caracterizar la dieta cetogénica como una alternativa en el tratamiento de la epilepsia refractaria en niños para mejorar su calidad de vida, con la consiguiente disminución de su costo económico. La epilepsia catastrófica o refractaria es un problema social de salud que la padecen pacientes pediátricos, generalmente en los primeros días de vida, de difícil control y con múltiples crisis pese al empleo de fármacos antiepilépticos en dosis tóxica, e incluye diferentes síndromes y enfermedades con características propias. Esta dieta ofrece una esperanza para aquellos pacientes que se resisten al tratamiento con medicamentos y su solución representa un valor social porque su beneficio supera el costo.

A descriptive study with objective to characterize the ketogenic diet as alternative in the treatment of the refractory epilepsy in children for elevating their quality of life, with the consequent decreasing of its economic cost, was carried out. The catastrophic or refractory epilepsy is one health social problem that the pediatric patient suffers, generally in the first days of life, the hard control and with numerous crisis despite of the use of antiepileptic drugs in toxic doses, and include different syndromes and diseases with own characteristics. This diet offers one hope, and its solution represents a social value because the benefit overcomes the cost for those patients that resist the drug treatment.