Fluorescence in situ hybridization for detection of classical propionibacteria with specific 16S rRNA-targeted probes and its application to enumeration in Gruyère cheese

The classical or dairy propionibacteria have well-documented industrial applications and have been proposed for probiotic applications. Given their industrial importance it is necessary to employ fast and reliable techniques to monitor the growth during products elaboration, industrial fermentations or the intestinal transit. Therefore, the aim of this investigation was to design oligonucleotide probes targeting the 16S rRNA of dairy propionibacteria and optimise the fluorescence in situ hybridization (FISH) protocol to detect these bacteria. Two specific probes were in silico designed to detect Propionibacterium freudenreichii and P. jensenii, named Pfr435 and Pj446 respectively. The FISH protocol was optimised for the hybridisation of propionibacteria cells with the universal probe Eub338 and the designed probes. These probes were assayed in situ for their specificity to hybridise species of propionibacteria by observation using fluorescence microscopy and results were compared with the probe Pap446 previously designed for P. acidipropionici. Probes Pap446, Pfr435 and Pj446 were also evaluated by fluorescence spectrophotometry to assess the influence of cells physiological state during growth in batch culture in the fluorescence intensity. The maximum fluorescence intensity was observed at the onset of the stationary phase of growth and was then reduced. However, changes on the cells permeability did not reduce the efficiency of 16S rRNA hybridisation with the fluorescence-labelled probes. Propionibacteria counts obtained by FISH and plate count methods were compared in a commercial Gruyère cheese. The results showed that this method can be used as a rapid technique for the enumeration of these bacteria in cheese samples.     

Reliability of automatic eddy-current equipment with a rotating magnetic field

This paper presents a method for estimating the reliability of eddy-current nondestructive inspection by using the rotating magnetic field method. The results of the inspection are considered as memory-less stochastic processes, with the possibility of representation in a bi-dimensional space: ‘good product’ and ‘bad product’. The general formulae are calculated by using the master equation for the time behaviour of the probabilities, the numerical data used for results estimation being obtained by means of an automatic eddy current installation with rotating magnetic field. The samples used had 20 μm, 40 μm and 98 μm deep artificial faults, and the control speed ranged between 0.05 and 1 m s⁻¹.     

A holonic architecture for the global road transportation system

In recent years the development of automated traffic systems has been gaining increasing attention, and a substantial amount of effort has been invested in trying to find a solution to problems associated to road transport. Among these problems are road accidents caused by human-related factors, such as tiredness, loss of control, a slow reaction time, limited field of view, etc. Another transport-related problem is that of loss of time which may be caused by a slow driving speed due to weather conditions, road conditions, visibility, traffic congestion, etc. In this paper, we present a distributed architecture and the underline distributed algorithm for solving the global road transportation system, which is being developed by several European Universities.     

Synthesis, characterization, and Ni(II) ion sorption properties of poly(styrene-co-divinylbenzene) functionalized with aminophosphonic acid groups

In this study, novel chelating resins containing aminophosphonic acid groups grafted on poly(styrene-co-divinylbenzene) (1 % or 15 % DVB) were obtained by “one-pot” reactions. The phosphorus contents were determined in order to calculate the degree of functionalization with pendant aminophosphonic acid groups. The fraction of repetitive units functionalized with aminophosphonic acid groups was determined by accepting the statistical structure of the repetitive unit of initial and final copolymer. These materials were characterized by FT-IR spectroscopy, EDX, and TGA. Adsorption properties of aminophosphonic acids grafted on poly(styrene-co-divinylbenzene) were examined for the removal of Ni(II) ions from aqueous solutions using the batch equilibrium procedure. By applying the kinetic models to the experimental data it was found that the adsorption of Ni(II) on the studied chelating resins is described by the pseudo-second-order kinetic model. For all the studied systems, Langmuir isotherm model provided an excellent fit of the equilibrium adsorption data.     

Effects of diet on casein and fatty acid profiles of milk from goats differing in genotype for αS1-casein synthesis

This study investigated the interactions between nutrition and the genotype at α_S1-CN loci (CSN1S1) in goats, evaluating the impact of fresh forage-based diets and an energy supplement on the casein and fatty acid (FA) profiles of milk from Girgentana goats. Twelve goats were selected for having the same genotype at the α_S2-CN, β-CN, and κ-CN loci and differing in the CSN1S1 genotype: homozygous for strong alleles (AA) or heterozygous for strong and weak alleles (AF). Goats of each genotype were divided into three groups and, according to a 3 × 3 Latin square design, fed ad libitum three diets: sulla fresh forage (SFF), SFF plus 800 g/day of barley (SFB), and mixed hay plus 800 g/day of barley (MHB). The SFB diet led to higher-energy intake and milk yield. The energy-supplemented diets (SFB, MHB) reduced milk fat and urea and increased coagulation time. The fresh forage diets (SFF, SFB) increased dry matter (DM) and crude protein (CP) intake and milk β-CN. Diet had a more pronounced effect than CSN1S1 genotype on milk FA profile, which was healthier from goats fed the SFF diet, due to the higher content of rumenic acid, polyunsaturated, and omega-3 FAs. The AA milk had longer coagulation time and higher curd firmness, higher short- and medium-chain FAs (SMFA), and lower oleic acid than AF milk. Significant diet by genotype interactions indicated the higher milk yield of AA goats than AF goats with the higher-energy SFB diet and the lower synthesis of SMFA in AF than in AA goats with the SFF diet.     

Molecular detection of pathogens in water - The pros and cons of molecular techniques

Pollution of water by sewage and run-off from farms produces a serious public health problem in many countries. Viruses, along with bacteria and protozoa in the intestine or in urine are shed and transported through the sewer system. Even in highly industrialized countries, pathogens, including viruses, are prevalent throughout the environment. Molecular methods are used to monitor viral, bacterial, and protozoan pathogens, and to track pathogen- and source-specific markers in the environment. Molecular techniques, specifically polymerase chain reaction-based methods, provide sensitive, rapid, and quantitative analytical tools with which to study such pathogens, including new or emerging strains. These techniques are used to evaluate the microbiological quality of food and water, and to assess the efficiency of virus removal in drinking and wastewater treatment plants. The range of methods available for the application of molecular techniques has increased, and the costs involved have fallen. These developments have allowed the potential standardization and automation of certain techniques. In some cases they facilitate the identification, genotyping, enumeration, viability assessment, and source-tracking of human and animal contamination. Additionally, recent improvements in detection technologies have allowed the simultaneous detection of multiple targets in a single assay. However, the molecular techniques available today and those under development require further refinement in order to be standardized and applicable to a diversity of matrices. Water disinfection treatments may have an effect on the viability of pathogens and the numbers obtained by molecular techniques may overestimate the quantification of infectious microorganisms. The pros and cons of molecular techniques for the detection and quantification of pathogens in water are discussed.     

All-in-One: Multifunctional Hydrogel Accelerates Oxidative Diabetic Wound Healing through Timed-Release of Exosome and Fibroblast Growth Factor

The treatment of diabetic wounds remains a major challenge in clinical practice, with chronic wounds characterized by multiple drug-resistant bacterial infections, angiopathy, and oxidative damage to the microenvironment. Herein, a novel in situ injectable HA@MnO₂/FGF-2/Exos hydrogel is introduced for improving diabetic wound healing. Through a simple local injection, this hydrogel is able to form a protective barrier covering the wound, providing rapid hemostasis and long-term antibacterial protection. The MnO₂/ε-PL nanosheet is able to catalyze the excess H₂O₂ produced in the wound, converting it to O₂, thus not only eliminating the harmful effects of H₂O₂ but also providing O₂ for wound healing. Moreover, the release of M2-derived Exosomes (M2 Exos) and FGF-2 growth factor stimulates angiogenesis and epithelization, respectively. These in vivo and in vitro results demonstrate accelerated healing of diabetic wounds with the use of the HA@MnO₂/FGF-2/Exos hydrogel, presenting a viable strategy for chronic diabetic wound repair.     

A location-based policy-specification language for mobile devices

The dramatic rise in mobile applications has greatly increased threats to the security and privacy of users. Security mechanisms on mobile devices are currently limited, so users need more expressive ways to ensure that downloaded mobile applications do not act maliciously. Policy-specification languages were created for this purpose; they allow the enforcement of user-defined policies on third-party applications. We have implemented LoPSiL, a location-based policy-specification language for mobile devices. This article describes LoPSiL’s design and implementation, several example policies, and experiments that demonstrate LoPSiL’s viability for enforcing policies on mobile devices.     

The Globus Replica Location Service: Design and Experience

Distributed computing systems employ replication to improve overall system robustness, scalability, and performance. A Replica Location Service (RLS) offers a mechanism to maintain and provide information about physical locations of replicas. This paper defines a design framework for RLSs that supports a variety of deployment options. We describe the RLS implementation that is distributed with the Globus Toolkit and is in production use in several Grid deployments. Features of our modular implementation include the use of soft-state protocols to populate a distributed index and Bloom filter compression to reduce overheads for distribution of index information. Our performance evaluation demonstrates that the RLS implementation scales well for individual servers with millions of entries and up to 100 clients. We describe the characteristics of existing RLS deployments and discuss how RLS has been integrated with higher-level data management services.