Occurrence of Listeria spp. in critical control points and the environment of Minas Frescal cheese processing

Critical control points (CCPs) associated with Minas Frescal cheese (a Brazilian soft white cheese, eaten fresh) processing in two dairy factories were determined using flow diagrams and microbiological tests for detection of Listeria monocytogenes and other species of Listeria. A total of 218 samples were collected along the production line and environment. The CCPs identified were reception of raw milk, pasteurization, coagulation and storage. Thirteen samples were positive for Listeria; 9 samples were Listeria innocua, 2 were Listeria grayi and 2 were L. monocytogenes. In factory A, Listeria was found in 50% of raw milk samples, 33.3% of curd samples, 16.7% of pasteurized milk samples, 16.7% of cheese samples and 25% of rubber pipes used to transport the whey. The microorganism was not obtained from environmental samples in this plant. In factory B, Listeria was found in one sample of raw milk (16.7%) and in three samples of environment (17.6%) and L. monocytogenes was obtained from raw milk (16.7%) and the floor of the cheese refrigeration room (14.3%). Two serotypes, 4b and 1/2a, were observed among the strains of L. monocytogenes isolated, both which are frequently involved in outbreaks of food-borne listeriosis and sporadic cases of the disease all over the world.     

Insights to the Structural Basis for the Stereospecificity of the Escherichia coli Phytase,AppA

AppA, the Escherichia coli periplasmic phytase of clade 2 of the histidine phosphatase (HP2) family, has been well-characterized and successfully engineered for use as an animal feed supplement. AppA is a 1D-6-phytase and highly stereospecific but transiently accumulates 1D-myo-Ins(2,3,4,5)P₄ and other lower phosphorylated intermediates. If this bottleneck in liberation of orthophosphate is to be obviated through protein engineering, an explanation of its rather rigid preference for the initial site and subsequent cleavage of phytic acid is required. To help explain this behaviour, the role of the catalytic proton donor residue in determining AppA stereospecificity was investigated. Four variants were generated by site-directed mutagenesis of the active site HDT amino acid sequence motif containing the catalytic proton donor, D304. The identity and position of the prospective proton donor residue was found to strongly influence stereospecificity. While the wild-type enzyme has a strong preference for 1D-6-phytase activity, a marked reduction in stereospecificity was observed for a D304E variant, while a proton donor-less mutant (D304A) displayed exclusive 1D-1/3-phytase activity. High-resolution X-ray crystal structures of complexes of the mutants with a non-hydrolysable substrate analogue inhibitor point to a crucial role played by D304 in stereospecificity by influencing the size and polarity of specificity pockets A and B. Taken together, these results provide the first evidence for the involvement of the proton donor residue in determining the stereospecificity of HP2 phytases and prepares the ground for structure-informed engineering studies targeting the production of animal feed enzymes capable of the efficient and complete dephosphorylation of dietary phytic acid.     

Polyurethane fibrous membranes tailored by rotary jet spinning for tissue engineering applications

Polymeric membranes have gained popularity as fibrous structures for tissue regeneration. This research focuses on the rotary jet spinning (RJS) process combined with a polymer as a strategy for designing membranes. To this end, RJS-polyurethane (RJS-PU) membranes with different microstructures were produced. Considering the effects of solution properties on fiber production, the viscosity of PU solutions was evaluated. Membrane morphology was studied based on scanning electron microscopy and 2D fast Fourier transform analysis. The chemical and thermal properties were characterized by Fourier-transform infrared spectroscopy and thermogravimetric analysis, respectively. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and Live/Dead cell assays were performed to determine the material cytotoxicity by assessment of the profile of proliferation and cell viability. The results indicated that the combination of PU and RJS was an effective one for the production of fibrous structures for tissue engineering applications, demonstrating good compatibility with the cultured osteoblastic cell line. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020 , 137, 48455.     

Comparative Analysis of Phenolic Composition of Six Commercially Available Chamomile (Matricaria chamomilla L.) Extracts: Potential Biological Implications

Several phytochemical-containing herbal extracts are increasingly marketed as health-promoting products. In particular, chamomile (Matricaria recutita L.) is well known for its anti-inflammatory, analgesic, and antitumor properties. Here, we evaluated differences in chemical composition among six commercially available products and their potential impact on biological activity in human immortalized colonocytes. Our investigation encompassed: (i) preparation of dry extracts and yield evaluation; (ii) qualitative and quantitative analysis of phenol content; (iii) modulation of redox state; and (iv) bioavailability of main bioactive compounds. We demonstrated that apparently identical products showed huge heterogeneity, in terms of yield extraction, chemical composition, and antioxidant effects. All samples contained high amounts of flavonoids and cinnamic acid derivatives, but differentially concentrated in the six extracts. Depending on polyphenol content, chamomile samples possessed variable antioxidant potential, in terms of decreased radical generation and increased reduced glutathione levels. The observed effects might be ascribed to flavones (apigenin, luteolin, and their glycones) highly represented in the six extracts. Nonetheless, chamomile extracts exerted cytotoxic effects at high concentrations, suggesting that a herbal medicine is not always safe. In conclusion, due to the complexity and variability of plant matrices, studies evaluating effectiveness of chamomile should always be accompanied by preliminary characterization of phytochemical composition.     

The physical and chemical composition of the lower mantle

This article reviews some of the recent advances made within the field of mineral physics. In order to link the observed seismic and density structures of the lower mantle with a particular mineral composition, knowledge of the thermodynamic properties of the candidate materials is required. Determining which compositional model best matches the observed data is difficult because of the wide variety of possible mineral structures and compositions. State-of-the-art experimental and analytical techniques have pushed forward our knowledge of mineral physics, yet certain properties, such as the elastic properties of lower mantle minerals at high pressures and temperatures, are difficult to determine experimentally and remain elusive. Fortunately, computational techniques are now sufficiently advanced to enable the prediction of these properties in a self-consistent manner, but more results are required.A fundamental question is whether or not the upper and lower mantles are mixing. Traditional models that involve chemically separate upper and lower mantles cannot yet be ruled out despite recent conflicting seismological evidence showing that subducting slabs penetrate deep into the lower mantle and that chemically distinct layers are, therefore, unlikely.Recent seismic tomography studies giving three-dimensional models of the seismic wave velocities in the Earth also base their interpretations on the thermodynamic properties of minerals. These studies reveal heterogeneous velocity and density anomalies in the lower mantle, which are difficult to reconcile with mineral physics data.     

Electrochemical Immuno-Vascular Solid State Point of Care Testing （POCT）

In this work, it is proposed a POCT and innovative method of immunoassay for the detection of C-reactive protein and IgG, using Amperometry coupled to solid state kit, connected to a micro-flow system with comparable sensitivity to a high sensitivity CRP ELISA （hsCRP） and IgG ELISA, with 1-3 min turnaround time to result. Samples of CRP （0 to 250 ng·mL-1） and IgG or diluted spiked human serum are injected through a solid state polymeric kit, micro-flow sensor channels of a SWCT SPE nano-modified biosensor. Preparing two kits immuno-assays, in the same micro-column, built on oxirane groups of polymeric bead surface, with biological support to CRP and IgG biomarkers recognition, in a real time scheme, at the end of analyte injection the initial rate of change in current intensity I/A was proportional to CRP respectively IgG concentration, with low detection limit （LOD） of 0.1 ng·mL-1. It was revealed that CRP/IgG concentrations in serum that might be expected in both normal and pathological conditions can be detected in a real-time-efficient, multi-immunoassay with solid state detection kit technology with determined CRP/lgG concentrations in close agreement with those determined using a commercially available high sensitivity EL1SA.     

A distributed framework for multi-risk assessment of natural hazards used to model the effects of forest fire on hydrology and sediment yield

Within the European Commission-funded MEDIGRID project, Grid computing technology is used to integrate various natural hazard models and data sets, maintained independently at different centres in Europe, into a single system, accessible to users over the internet. Each centre forms a process (application) or data storage node and has been fitted with the Globus toolkit, which provides the distributed computing environment functionality that is required for the system set up. In addition, several Grid data management components were developed to allow the system to operate on different computing platforms. Access to the data and application management services is enabled through a Grid Portal. A series of portlets enable users to access the system, providing a personalised interface to the Grid. Integration of the individual models required them to be modified as web services, so as to be run remotely over the internet. As the models have different data characteristics, a common data format was adopted for creating harmonised data sets and allowing the exchange of data between the models. As an example, the Fire Spread Engine model is used to derive a map of areas that have been burnt by fire. This forms an input to the SHETRAN hydrology, soil erosion and landslide model, which in turn could provide data for other models such as vegetation regeneration. The use of the system is demonstrated for a site in south-west Spain where a large forest fire occurred on 2 August 2003. The MEDIGRID system marks an advance in the integration of independently constructed models to provide improved hazard assessment technology.