Métodos de transducción usados en biosensores: amperometría y fluorescencia

RESUMEN: Los biosensores tienen aplicación en una gran variedad de campos, incluyendo el análisis ambiental, biomedicina, biodefensa, alimentación y agricultura, entre otros. En este tipo de sensores, un material biológico (conocido como biomediador) reacciona con el analito y un sistema de transducción apropiado transforma dicha reacción en una señal eléctrica que puede ser procesada, almacenada o transmitida usando sistemas electrónicos. En este artículo, se describen dos métodos de transducción usados en aplicaciones bio-sensoriales: la amperometria que consiste en la medida del transferimento electrónico (corriente) del biomediador y fluorescencia que es basada en la medida de la luz re-emitida. Se enfatiza en el diseño electrónico (selección de componentes, topologia de los circuitos, problemas comunes y soluciones). Estos diseños han sido utilizados en el desarrollo de instrumentos comerciales para biosensores, caracterizados por bajos costes de producción y portabilidad.ABSTRACT: Biosensor devices have applications in a variety of fields as environmental analysis, biomedical, bio-defense, food and agriculture. On this kind of sensors, a biological material (known as biomediator) reacts with target analytes and an appropriated transduction system converts that reaction to an electrical signal that can be processed, saved and transmitted by using electronic systems. In this article, two transduction methods used for biosensing applications are described: amperometry that is based on the measurement of the electron transfer occurring inside the biomediator and fluorescence, that is based on the measurement of the re-emitted light. Emphasis has been done on the electronics design, including component selection, useful circuit topologies and common problems and solutions. Electronics has been validated for the development of biosensor-based instruments characterized by low production costs and portability

Regeneration of plants from embryogenic callus-derived protoplasts of Garganega and Sangiovese grapevine (Vitis vinifera L.) cultivars

Protoplasts are useful research tools for basic and applied plant science, but the regeneration of whole plants from protoplasts is challenging in most of agronomically important crops, including grapevine (Vitis vinifera L.). Here we describe an efficient protocol for the induction of embryogenic callus, the isolation of protoplasts, and the regeneration of whole grapevine plants in two Italian grapevine cultivars. Embryogenic callus was induced successfully from stamens collected from immature flowers. Isolated protoplasts were tested to confirm their viability and then cultivated using the disc-culture method, at a density of 1\u2009 7\u2009105 protoplasts/mL in solid Nitsch\u2019s medium supplemented with 2 mg/L 1-naphthaleneacetic acid and 0.5 mg/L 6-benzylaminopurine. After 3\u20134 months, the protoplasts of both cultivars regenerated with similar efficiency into cotyledonal-stage somatic embryos. The somatic embryos were transferred to solid Nitsch\u2019s medium supplemented with 30 g/L sucrose and 2 g/L gellan gum, and were maintained in the dark for 4 weeks. This step was necessary for the embryo to complete germination, allowing subsequent shoot elongation in response to light on a medium with 4 \ub5M 6-benzylaminopurine. Then root elongation occurred after transferring on a medium with 0.5 \ub5M 1-naphthaleneacetic. After\u2009~\u20096 months from the isolation of protoplasts, normal plants were regenerated, which were moved to the greenhouse. The protoplasts could also be transfected using the polyethylene glycol method, as confirmed using a plasmid carrying the yellow florescent protein marker gene. The new method is therefore compatible with biotechnological applications such as gene transfer and genome editing

Constraints on planetary tidal dissipation from a detailed study of Kepler 91b

Context. With the detection of thousands of exoplanets, characterising their dynamical evolution in detail represents a key step in the understanding of their formation. Studying the dissipation of tides occurring both in the host star and in the planets is of great relevance in order to investigate the distribution of the angular momentum occurring among the objects populating the system and to studying the evolution of the orbital parameters. From a theoretical point of view, the dissipation of tides throughout a body may be studied by relying on the so-called phase or time-lag equilibrium tides model in which the reduced tidal quality factor Q'p, or equivalently the product between the love number and the time lag (k2DeltaT), describe how efficiently tides are dissipated within the perturbed body. Constraining these factors by looking at the current configuration of the exoplanetary system is extremely challenging, and simulations accounting for the evolution of the system as a whole might help to shed some light on the mechanisms governing this process. Aims. We aim to constrain the tidal dissipation factors of hot-Jupiter-like planets by studying the orbital evolution of Kepler-91b. Methods. We firstly carried out a detailed asteroseismc characterisation of Kepler-91 and computed a dedicated stellar model using both classical and astereoseismic constraints. We then coupled the evolution of the star to the one of the planets by means of our orbital evolution code and studied the evolution of the system by accounting for tides dissipated both in the planet and in the host star. Results. We found that the maximum value for k2DeltaT (or equivalently the minimum value for Q'p) determining the efficiency of equilibrium tides dissipation occurring within Kepler-91b is 0.4 pm 0.25 s (4.5+5.8 * 10^5).Comment: accepted for publication in Astronomy & Astrophysic

High prevalence of anti-hepatitis e virus antibodies among blood donors in central Italy, february to march 2014

Prevalence of anti-hepatitis E virus (HEV) antibodies is highly variable in developed countries, which seems partly due to differences in assay sensitivity. Using validated sensitive assays, we tested 313 blood donors attending a hospital transfusion unit in central Italy in January and February 2014 for anti-HEV IgG and IgM and HEV RNA. Data on HEV exposure were collected from all donors. Overall anti-HEV IgG prevalence was 49% (153/313). Eating raw dried pig-liver sausage was the only independent predictor of HEV infection (adjusted prevalence rate ratio = 2.14; 95% confidence interval: 1.23–3.74). Three donors were positive for either anti-HEV IgM (n = 2; 0.6%) or HEV RNA (n = 2; 0.6%); they were completely asymptomatic, without alanine aminotransferase (ALT) abnormalities. Of the two HEV RNA-positive donors (both harbouring genotype 3), one was anti-HEV IgG- and IgM-positive, the other was anti-HEV IgG- and IgM-negative. The third donor was positive for anti-HEV IgG and IgM but HEV RNA-negative. HEV infection is therefore hyperendemic among blood donors (80% men 18–64 years-old) from central Italy and associated with local dietary habits. Nearly 1% of donors have acute or recent infection, implying potential transmission to blood recipients. Neither ALT nor anti-HEV IgM testing seems useful to prevent transfusion-transmitted HEV infection. © 2016, European Centre for Disease Prevention and Control

The Mitochondrial A10398G Polymorphism, Interaction with Alcohol Consumption, and Breast Cancer Risk

Polymorphisms in the mitochondrial genome are hypothesized to be associated with risk of various diseases, including cancer. However, there has been conflicting evidence for associations between a common polymorphism in the mitochondrial genome (A10398G, G10398A in some prior reports) and breast cancer risk. Reactive oxygen species, a by-product of mitochondrial energy production, can lead to oxidative stress and DNA damage in both the mitochondria and their cells. Alcohol consumption, which may also lead to oxidative stress, is associated with breast cancer risk. Therefore, we hypothesized that polymorphisms in the mitochondrial genome interact with alcohol consumption to alter breast cancer risk. We genotyped the A10398G polymorphism in a case-control study nested within the Nurses' Health Study (NHS, 1,561 cases, 2,209 controls). We observed an interaction between alcohol consumption (yes/no) and A10398G on breast cancer risk (p-int = 0.03). The risk associated with alcohol consumption was limited to carriers of the 10398G allele (Odds Ratio 1.52, 95% Confidence Interval 1.10–2.08 comparing drinkers to non-drinkers). However, we were unable to replicate these findings in the Women's Health Study (WHS, 678 cases, 669 controls), although the power to detect this interaction in the WHS was low (power = 0.57). Further examination of this interaction, such as sufficiently powered epidemiological studies of cancer risk or associations with biomarkers of oxidative stress, may provide further evidence for GxE interactions between the A10398G mitochondrial polymorphism and alcohol consumption on breast cancer risk

Failure regime in (1+1) dimensions in fibrous materials

In this paper, we introduce a model for fracture in fibrous materials that takes into account the rupture height of the fibers, in contrast with previous models. Thus, we obtain the profile of the fracture and calculate its roughness, defined as the variance around the mean height. We investigate the relationship between the fracture roughness and the fracture toughness.Comment: 4 pages, 4 figures.eps, Revte

Is the Salmonella contamination of swine carcasses at slaughter related to the Salmonella load in caecum?

The aim of this study was to examine the relationship between the load of Salmonella spp. in caeca and the carcass contamination in an Italian slaughterhouse. The sampling scheme was designed to be representative of the pigs slaughtered in a day and to estimate a 12% prevalence of pigs highly contaminated by Salmonella spp. (HCP, cecal load ≥3log). Environmental swabs were taken before slaughter. Cecal contents and carcass swabs were collected from the same pig. Salmonella MPN were estimated according to ISO6579- 2:2012/A1 and ISO7218:2007/E. The overall Salmonella prevalence were 34.64% and 7.19% for ceca and carcasses respectively, with S. Derby and S. 4,[5],12:i:- being the prevalent serotypes. The HCP prevalence was 11.44%. 7/59 environmental swabs tested positive; when the same serotype was isolated from the environment and from carcasses, the samples were excluded from further analysis. Statistical analysis was performed to investigate the relationship between Salmonella spp. loads in the cecum and contamination of the carcass of the same pig and the prevalence of HCP and the contamination of carcasses on the same day. For this purpose, the days were classified as “high prevalence days” depending on the proportion of caeca resulted positive (≥36%) and as “high load” days depending on the prevalence of HCP (≥10%). A correlation between the contamination of carcasses and the cecal Salmonella loads of the same animal was found (Spearman’s correlation coefficient: 0.2254; p-value=0.0001). No correlation was found between the contamination of carcasses and the categorization of the day of sampling as “high prevalence day”. Conversely, a correlation was found between the contamination of carcasses and the “high load” category of the sampling day (Wilcoxon test, p=0.0011). Notably, not the prevalence of pigs carrying Salmonella spp. but the prevalence of highly contaminated pigs was shown to be related to the contamination of carcasses

Genetic heterogeneity of porcine enteric caliciviruses identified from diarrhoeic piglets

Enteric caliciviruses (noroviruses and sapoviruses) are responsible for the majority of non-bacterial gastroenteritis in humans of all age groups. Analysis of the polymerase and capsid genes has provided evidence for a huge genetic diversity, but the understanding of their ecology is limited. In this study, we investigated the presence of porcine enteric caliciviruses in the faeces of piglets with diarrhoea. A total of 209 samples from 118 herds were analyszd and calicivirus RNA was detected by RT-PCR in 68 sample (32.5%) and in 46 herds (38.9%), alone or in mixed infection with group A and C rotaviruses. Sequence and phylogenetic analysis of the calicivirus-positive samples characterized the majority as genogroup III (GGIII) sapoviruses. Unclassified caliciviruses, distantly related to the representatives of the other sapovirus genogroups, were identified in five herds, while one outbreak was associated with a porcine sapovirus related genetically to human GGII and GGIV sapovirus strains. By converse, norovirus strains were not detected. Altogether, these data suggest the epidemiological relevance of porcine enteric caliciviruses and suggest a role in the etiology of piglets diarrhoe