Different Involvement of Vimentin during Invasion by Listeria monocytogenes at the Blood–Brain and the Blood–Cerebrospinal Fluid Barriers In Vitro

The human central nervous system (CNS) is separated from the blood by distinct cellular barriers, including the blood–brain barrier (BBB) and the blood–cerebrospinal fluid (CFS) barrier (BCSFB). Whereas at the center of the BBB are the endothelial cells of the brain capillaries, the BCSFB is formed by the epithelium of the choroid plexus. Invasion of cells of either the BBB or the BCSFB is a potential first step during CNS entry by the Gram-positive bacterium Listeria monocytogenes (Lm). Lm possesses several virulence factors mediating host cell entry, such as the internalin protein family—including internalin (InlA), which binds E-cadherin (Ecad) on the surface of target cells, and internalin B (InlB)—interacting with the host cell receptor tyrosine kinase Met. A further family member is internalin (InlF), which targets the intermediate filament protein vimentin. Whereas InlF has been shown to play a role during brain invasion at the BBB, its function during infection at the BCSFB is not known. We use human brain microvascular endothelial cells (HBMEC) and human choroid plexus epithelial papilloma (HIBCPP) cells to investigate the roles of InlF and vimentin during CNS invasion by Lm. Whereas HBMEC present intracellular and surface vimentin (besides Met), HIBCPP cells do not express vimentin (except Met and Ecad). Treatment with the surface vimentin modulator withaferin A (WitA) inhibited invasion of Lm into HBMEC, but not HIBCPP cells. Invasion of Lm into HBMEC and HIBCPP cells is, however, independent of InlF, since a deletion mutant of Lm lacking InlF did not display reduced invasion rates.     

The Assessment of Activity of Antiseptic Agents against Biofilm of Staphylococcus aureus Measured with the Use of Processed Microscopic Images

Staphylococcal biofilms are major causative factors of non-healing wound infections. Their treatment algorithms recommend the use of locally applied antiseptic agents to counteract the spread of infection. The efficacy of antiseptics against biofilm is assessed in vitro by a set of standard quantitative and semi-quantitative methods. The development of software for image processing additionally allowed for the obtainment of quantitative data from microscopic images of biofilm dyed with propidium iodine and SYTO-9 reagents, differentiating dead cells from live ones. In this work, the method of assessment of the impact of antiseptic agents on staphylococcal biofilm in vitro, based on biofilms’ processed images, was proposed and scrutinized with regard to clinically relevant antiseptics, polyhexanide, povidone–iodine and hypochlorite. The standard quantitative culturing method was applied to validate the obtained data from processed images. The results indicated significantly higher activity of polyhexanide and povidone–iodine than hypochlorite against staphylococcal biofilm. Taking into account the fact that in vitro results of the efficacy of antiseptic agents against staphylococcal biofilm are frequently applied to back up their use in hospitals and ambulatory units, our work should be considered an important tool; providing reliable, quantitative data in this regard.     

Aldehyde Accumulation in Aged Alcoholic Beer: Addressing Acetaldehyde Impacts on Upper Aerodigestive Tract Cancer Risks

Aldehydes, particularly acetaldehyde, are carcinogenic molecules and their concentrations in foodstuffs should be controlled to avoid upper aerodigestive tract (UADT) and liver cancers. Highly reactive, acetaldehyde forms DNA and protein adducts, impairing physiological functions and leading to the development of pathological conditions. The consumption of aged beer, outside of the ethanol metabolism, exposes habitual drinkers to this carcinogen, whose concentrations can be over-increased due to post-brewing chemical and biochemical reactions. Storage-related changes are a challenge faced by the brewing industry, impacting volatile compound formation and triggering flavor instability. Aldehydes are among the volatile compounds formed during beer aging, recognized as off-flavor compounds. To track and understand aldehyde formation through multiple pathways during beer storage, consequent changes in flavor but particularly quality losses and harmful compound formation, this systematic review reunited data on volatile compound profiles through gas chromatography analyses from 2011 to 2021. Conditions to avoid flavor instability and successful methods for reducing beer staling, and consequent acetaldehyde accumulation, were raised by exploring the dynamic conversion between free and bound-state aldehydes. Future research should focus on implementing sensory analyses to investigate whether adding aldehyde-binding agents, e.g., cysteine and bisulfite, would contribute to consumer acceptance, restore beer flavor, and minimize acetaldehyde-related health damage.     

Applications of Metabolomics in Calcium Metabolism Disorders in Humans

The pathogenesis of the disorders of calcium metabolism is not fully understood. This review discusses the studies in which metabolomics was applied in this area. Indeed, metabolomics could play an essential role in discovering biomarkers and elucidating pathological mechanisms. Despite the limited bibliography, the present review highlights the potential of metabolomics in identifying the biomarkers of some of the most common endocrine disorders, such as primary hyperparathyroidism (PHPT), secondary hyperparathyroidism (SHPT), calcium deficiency, osteoporosis and vitamin D supplementation. Metabolites related to above-mentioned diseorders were grouped into specific classes and mapped into metabolic pathways. Furthermore, disturbed metabolic pathways can open up new directions for the in-depth exploration of the basic mechanisms of these diseases at the molecular level.     

Application Level Fault Tolerance in Heterogeneous Networks of Workstations

We have explored methods for checkpointing and restarting processes within the distributed object migration environment (Dome), a C++ library of data parallel objects that are automatically distributed over heterogeneous networks of workstations (NOWs). System level checkpointing methods, although transparent to the user, were rejected because they lack support for heterogeneity. We have implemented application level checkpointing which places the checkpoint and restart mechanisms within Dome's C++ objects. Application level checkpointing has been implemented with a library-based technique for the programmer and a more transparent preprocessor-based technique. Dome's implementation of checkpointing successfully checkpoints and restarts processes on different numbers of machines and different architectures. Results from executing Dome programs across a NOW with realistic failure rates have been experimentally determined and are compared with theoretical results. The overhead of checkpointing is found to be low, while providing substantial decreases in expected runtime on realistic systems.     

Issues in the Reconstruction of Gene Order Evolution

As genomes evolve over hundreds of millions years, the chromosomes become rearranged, with segments of some chromosomes inverted, while other chromosomes reciprocally exchange chunks from their ends. These rearrangements lead to the scrambling of the elements of one genome with respect to another descended from a common ancestor. Multidisciplinary work undertakes to mathematically model these processes and to develop statistical analyses and mathematical algorithms to understand the scrambling in the chromo...     

Approximation Algorithms for <Emphasis Type="Italic">k</Emphasis>-hurdle Problems

The polynomial-time solvable k-hurdle problem is a natural generalization of the classical s-t minimum cut problem where we must select a minimum-cost subset S of the edges of a graph such that |p∩S|≥k for every s-t path p. In this paper, we describe a set of approximation algorithms for “k-hurdle” variants of the NP-hard multiway cut and multicut problems. For the k-hurdle multiway cut problem with r terminals, we give two results, the first being a pseudo-approximation algorithm that outputs a (k−1)-hurdle solution whose cost is at most that of an optimal solution for k hurdles. Secondly, we provide a 2(1-\frac1r)2(1-\frac{1}{r})-approximation algorithm based on rounding the solution of a linear program, for which we give a simple randomized half-integrality proof that works for both edge and vertex k-hurdle multiway cuts that generalizes the half-integrality results of Garg et al. for the vertex multiway cut problem. We also describe an approximation-preserving reduction from vertex cover as evidence that it may be difficult to achieve a better approximation ratio than 2(1-\frac1r)2(1-\frac{1}{r}). For the k-hurdle multicut problem in an n-vertex graph, we provide an algorithm that, for any constant ε>0, outputs a ⌈(1−ε)k⌉-hurdle solution of cost at most O(log n) times that of an optimal k-hurdle solution, and we obtain a 2-approximation algorithm for trees.