Replication-Deficient Lymphocytic Choriomeningitis Virus-Vectored Vaccine Candidate for the Induction of T Cell Immunity against Mycobacterium tuberculosis

Mycobacterium tuberculosis (Mtb) represents a major burden to global health, and refined vaccines are needed. Replication-deficient lymphocytic choriomeningitis virus (rLCMV)-based vaccine vectors against cytomegalovirus have proven safe for human use and elicited robust T cell responses in a large proportion of vaccine recipients. Here, we developed an rLCMV vaccine expressing the Mtb antigens TB10.4 and Ag85B. In mice, rLCMV elicited high frequencies of polyfunctional Mtb-specific CD8 and CD4 T cell responses. CD8 but not CD4 T cells were efficiently boosted upon vector re-vaccination. High-frequency responses were also observed in neonatally vaccinated mice, and co-administration of rLCMV with Expanded Program of Immunization (EPI) vaccines did not result in substantial reciprocal interference. Importantly, rLCMV immunization significantly reduced the lung Mtb burden upon aerosol challenge, resulting in improved lung ventilation. Protection was associated with increased CD8 T cell recruitment but reduced CD4 T cell infiltration upon Mtb challenge. When combining rLCMV with BCG vaccination in a heterologous prime-boost regimen, responses to the rLCMV-encoded Mtb antigens were further augmented, but protection was not significantly different from rLCMV or BCG vaccination alone. This work suggests that rLCMV may show utility for neonatal and/or adult vaccination efforts against pulmonary tuberculosis.     

The Combination of Intestinal Alkaline Phosphatase Treatment with Moderate Physical Activity Alleviates the Severity of Experimental Colitis in Obese Mice via Modulation of Gut Microbiota,Attenuation of Proinflammatory Cytokines,Oxidative Stress Biomarkers and DNA Oxidative Damage in Colonic Mucosa

Inflammatory bowel diseases (IBD) are commonly considered as Crohn’s disease and ulcerative colitis, but the possibility that the alterations in gut microbiota and oxidative stress may affect the course of experimental colitis in obese physically exercising mice treated with the intestinal alkaline phosphatase (IAP) has been little elucidated. Mice fed a high-fat-diet (HFD) or normal diet (ND) for 14 weeks were randomly assigned to exercise on spinning wheels (SW) for 7 weeks and treated with IAP followed by intrarectal administration of TNBS. The disease activity index (DAI), grip muscle strength test, oxidative stress biomarkers (MDA, SOD, GSH), DNA damage (8-OHdG), the plasma levels of cytokines IL-2, IL-6, IL-10, IL-12p70, IL-17a, TNF-α, MCP-1 and leptin were assessed, and the stool composition of the intestinal microbiota was determined by next generation sequencing (NGS). The TNBS-induced colitis was worsened in obese sedentary mice as manifested by severe colonic damage, an increase in DAI, oxidative stress biomarkers, DNA damage and decreased muscle strength. The longer running distance and weight loss was observed in mice given IAP or subjected to IAP + SW compared to sedentary ones. Less heterogeneous microbial composition was noticed in sedentary obese colitis mice and this effect disappeared in IAP + SW mice. Absence of Alistipes, lower proportion of Turicibacter, Proteobacteria and Faecalibacterium, an increase in Firmicutes and Clostridium, a decrease in oxidative stress biomarkers, 8-OHdG content and proinflammatory cytokines were observed in IAP + SW mice. IAP supplementation in combination with moderate physical activity attenuates the severity of murine colitis complicated by obesity through a mechanism involving the downregulation of the intestinal cytokine/chemokine network and oxidative stress, the modulation of the gut microbiota and an improvement of muscle strength.     

Sprouted wheat grain with ferritin overexpression as a potential source of iron for cereal product fortification

Grains of winter wheat (Korweta variety) were partially germinated in an abiotic stress condition (0–15 mM FeSO₄ solutions) in order to accumulate iron by ferritin overexpression. The physical, chemical, biochemical properties and technological parameters of the obtained material as well as their resistance to an insect storage pest—granary weevil—were analyzed. Sprouted wheat grains with overexpression of ferritin contain 52 times more iron, more than 50 % in form of ferritin. The contents of reducing sugars, crude protein, fiber and ash as well as amylolytic activity increased significantly in sprouted wheat grain, while starch, fat, wet gluten, falling number and inhibition activity against granary weevil α-amylase decreased. Germinated wheat grain enriched with iron strongly affected granary weevil developmental parameters. Despite the fact that technological parameters of the product were deteriorated, it is worth to consider this material application as an iron supplement for cereal products, when the application of soybean is impossible.     

Freshness and quality attributes of cold stored Atlantic bonito (Sarda sarda) gravad

Quality and safety parameters of Atlantic bonito gravad during 42 days of storage at 7 ± 1 °C were analysed using monitoring sensory quality, microbial contamination, nucleotide degradation products, biogenic amines (BA), trimethylamine (TMA) and thiobarbituric acid reactive substances (TBARS). The shelf‐life of vacuum packed Atlantic bonito gravad was found to be 28 days. Total viable count (TVC) did not exceed 6 log CFU g^?1. Malonodialdehyde (MDA) content did not surpass 10 mg kg^?1. Nine biogenic amines were detected. Histamine level at the end of the storage was 355.3 mg kg^?1. TMA increased from 53.9 to 619.1 mg kg^?1. Among indicators based on the products of nucleotide degradation, the H value is best correlated with the sensory assessment. It can be concluded that the storage temperature of bonito gravad must be below 7 °C to prevent the threat to safety and quality posed by biogenic amines, especially histamine.     

Functional analysis of a rickettsial OmpA homology domain of Shigella flexneri icsA

Shigella flexneri is a gram-negative bacterium that causes diarrhea and dysentery by invasion and spread through the colonic epithelium. Bacteria spread by assembling actin and other cytoskeletal proteins of the host into "actin tails" at the bacterial pole; actin tail assembly provides the force required to move bacteria through the cell cytoplasm and into adjacent cells. The 120-kDa S. flexneri outer membrane protein IcsA is essential for actin assembly. IcsA is anchored in the outer membrane by a carboxy-terminal domain (the beta domain), such that the amino-terminal 706 amino acid residues (the alpha domain) are exposed on the exterior of the bacillus. The alpha domain is therefore likely to contain the domains that are important to interactions with host factors. We identify and characterize a domain of IcsA within the alpha domain that bears significant sequence similarity to two repeated domains of rickettsial OmpA, which has been implicated in rickettsial actin tail formation. Strains of S. flexneri and Escherichia coli that carry derivatives of IcsA containing deletions within this domain display loss of actin recruitment and increased accessibility to IcsA-specific antibody on the surface of intracytoplasmic bacteria. However, site-directed mutagenesis of charged residues within this domain results in actin assembly that is indistinguishable from that of the wild type, and in vitro competition of a polypeptide of this domain fused to glutathione S-transferase did not alter the motility of the wild-type construct. Taken together, our data suggest that the rickettsial homology domain of IcsA is required for the proper conformation of IcsA and that its disruption leads to loss of interactions of other IcsA domains within the amino terminus with host cytoskeletal proteins.     

Factors affecting the potentiometric response of all-solid-state solvent polymeric membrane calcium-selective electrode for low-level measurements

An all-solid-state calcium-selective electrode was constructed with poly(pyrrole) solid-contact doped with calcium complexing ligand Tiron. The potentiometric response of this sensor can have a linear range down to 10(-)(9) M with a slope close to Nernstian and detection limit equal to 10(-)(9.6). The effects of pH and the activity of the interfering ion in the conditioning solution on the potentiometric behavior of the constructed sensors were examined. Potential stability, reproducibility, and impedance studies were performed. The selectivity of the constructed electrode is better than that of the conventional calcium-selective electrode with internal filling solution of 10(-)(2) M CaCl(2) and comparable to that of the best liquid-contact electrodes.     

Strong Negation and Equivalence in the Safe Belief Semantics

The safe belief semantics uses intermediate logics to definean extension of answer sets to all propositional formulas, butonly considering one kind of negation. In this work we extendsafe beliefs adding the strong negation connective. The mainfeature of our extension is that strong negation can occur beforeany formula, and not only at the atomic level. We give resultsconcerning the relation between strong negation extensions ofintermediate logics and safe beliefs and consider the way inwhich strong negation can be eliminated from any formula whilepreserving its semantics. We also propose two new notions ofequivalence: substitution equivalence and contextualized equivalence.We prove that they are both more general than strong equivalenceand, for propositional formulas where strong negation may occurat the non-atomic level, substitution equivalence captures anotion of equivalence that cannot be captured by strong equivalencealone.