In vitro screening of probiotics and synbiotics according to anti-inflammatory and anti-proliferative effects

There is emerging evidence of the efficiency of probiotic, prebiotic and synbiotic treatments in inflammatory bowel diseases (IBDs) and one of their long-term complications, colorectal cancer (CRC). In this study, various strains of probiotic lactic acid bacteria, prebiotic glucooligosaccharides (GOS) or a synbiotic combination of the two were screened for anti-inflammatory and anti-proliferative effects in different in vitro models in the context of such diseases. To mimic IBD response to Gram negative bacteria, HT-29 cells were sensitised to inflammatory response to lipopolysaccharide (LPS) by IFNγ which increased expression of TLR4, the LPS biosensor, and were then treated by probiotics, prebiotics and synbiotics. Secreted IL-8 and activated NF-κB were monitored as inflammation biomarkers. A selection of active strains were then subjected to a second inflammatory cell culture model consisting of inflammatory activated transgenic Caco-2 cells transfected by a reporter gene under the control of NF-κB inducible promoter. Quantification of reporter gene expression allowed us to demonstrate some probiotic inhibitory properties or to confirm such characteristics in two different models. Proliferation of cancerous HT-29 cells was monitored by XTT assay. Only three probiotic strains induced a proliferation decrease, but with a lack of reproducibility. Binary or ternary probiotic associations, complemented or not by prebiotic GOS, significantly decreased proliferation, especially with a synbiotic association of Bifidobacterium breve, Lactococcus lactis and oligoalternan, a GOS. This combination was selected for the following experiments. We showed the involvement of both bacterial and carbohydrate compounds of this synbiotic in the observed effect by dose range tests. We demonstrated that this decrease in proliferation may be due to an induction of a differentiated phenotype, as shown by the up-regulation of intestinal alkaline phosphatase, a biomarker of differentiation, monitored by real-time RT-PCR in HT-29 cells treated by the selected synbiotics. Thus, this study demonstrates the ability of probiotics to exert anti-inflammatory effects and shows some anti-proliferative characteristics for a specific synbiotics. These products should be further evaluated in animal models to confirm the in vitro results.     

Food processing increases casein resistance to simulated infant digestion

The objective of this study was to determine whether processing could modify the resistance of casein (CN) to digestion in infants. A range of different dairy matrices was manufactured from raw milk in a pilot plant and subjected to in vitro digestion using an infant gut model. Digestion products were identified using MS and immunochemical techniques. Results obtained showed that CNs were able to resist digestion, particularly κ‐ and αs₂‐CN. Resistant areas were identified and corresponded to fragments hydrophobic at pH 3.0 (gastric conditions) and/or carrying post‐translational modifications (phosphorylation and glycosylation). Milk processing led to differences in peptide patterns and heat treatment of milk tended to increase the number of peptides found in digested samples. This highlights the likely impact of milk processing on the allergenic potential of CNs.     

Analysis of Integrin αIIb Subunit Dynamics Reveals Long-Range Effects of Missense Mutations on Calf Domains

Integrin α_IIbβ₃, a glycoprotein complex expressed at the platelet surface, is involved in platelet aggregation and contributes to primary haemostasis. Several integrin α_IIbβ₃ polymorphisms prevent the aggregation that causes haemorrhagic syndromes, such as Glanzmann thrombasthenia (GT). Access to 3D structure allows understanding the structural effects of polymorphisms related to GT. In a previous analysis using Molecular Dynamics (MD) simulations of α_IIb Calf-1 domain structure, it was observed that GT associated with single amino acid variation affects distant loops, but not the mutated position. In this study, experiments are extended to Calf-1, Thigh, and Calf-2 domains. Two loops in Calf-2 are unstructured and therefore are modelled expertly using biophysical restraints. Surprisingly, MD revealed the presence of rigid zones in these loops. Detailed analysis with structural alphabet, the Proteins Blocks (PBs), allowed observing local changes in highly flexible regions. The variant P741R located at C-terminal of Calf-1 revealed that the Calf-2 presence did not affect the results obtained with isolated Calf-1 domain. Simulations for Calf-1 + Calf-2, and Thigh + Calf-1 variant systems are designed to comprehend the impact of five single amino acid variations in these domains. Distant conformational changes are observed, thus highlighting the potential role of allostery in the structural basis of GT.     

COVID-19 Pandemic: Escape of Pathogenic Variants and MHC Evolution

We propose a new hypothesis that explains the maintenance and evolution of MHC polymorphism. It is based on two phenomena: the constitution of the repertoire of naive T lymphocytes and the evolution of the pathogen and its impact on the immune memory of T lymphocytes. Concerning the latter, pathogen evolution will have a different impact on reinfection depending on the MHC allomorph. If a mutation occurs in a given region, in the case of MHC allotypes, which do not recognize the peptide in this region, the mutation will have no impact on the memory repertoire. In the case where the MHC allomorph binds to the ancestral peptides and not to the mutated peptide, that individual will have a higher chance of being reinfected. This difference in fitness will lead to a variation of the allele frequency in the next generation. Data from the SARS-CoV-2 pandemic already support a significant part of this hypothesis and following up on these data may enable it to be confirmed. This hypothesis could explain why some individuals after vaccination respond less well than others to variants and leads to predict the probability of reinfection after a first infection depending upon the variant and the HLA allomorph.     

A replenishment policy based on joint optimization in a downstream pharmaceutical supply chain: centralized vs. decentralized replenishment

This paper considers a number of problems in a pharmaceutical downstream supply chain under specific constraints related to pharmaceutical products (e.g., their expiry date and regulations) and to inventory control (e.g., low prices of products, gift products or prices near zero, shortages prohibited but excess inventory allowed). As traditional models of inventory control and replenishment cannot optimize the total cost of the system, it is very important to consider the transportation cost as well. In this paper, we take into account some of these constraints and propose two models for multi-product replenishment policies, namely centralized and decentralized models. We seek to identify the best quantity and period of replenishment of products for a joint optimization of inventory and transportation costs. The proposed models can be applied to a specific family of products with a stable demand and high turnover rate, low prices (or gift products for some of them), and without any shortage. These two models are compared and the global approach is illustrated by a numerical example taken from a real-case study.     

The evolution of the exhaust diesel soot adsorptive properties after NO2 exposure and according to their SOF content

Several samples of exhaust diesel soot are investigated by inverse gas chromatography and linear solvation energy relationship (LSER) modelling according to their soluble organic fraction content and their time of exposure in oxidative conditions. The results demonstrate the evolution of the adsorptive properties of the studied materials towards volatile compounds during the oxidation under NO₂.     

Incorporating a Polyethyleneglycol Linker to Enhance the Hydrophilicity of Mitochondria-Targeted Triphenylphosphonium Constructs

The targeting of bioactive molecules and probes to mitochondria can be achieved by coupling to the lipophilic triphenyl phosphonium (TPP) cation, which accumulates several hundred-fold within mitochondria in response to the mitochondrial membrane potential (Δψ_m). Typically, a simple alkane links the TPP to its “cargo”, increasing overall hydrophobicity. As it would be beneficial to enhance the water solubility of mitochondria-targeted compounds we explored the effects of replacing the alkyl linker with a polyethylene glycol (PEG). We found that the use of PEG led to compounds that were readily taken up by isolated mitochondria and by mitochondria inside cells. Within mitochondria the PEG linker greatly decreased adsorption of the TPP constructs to the matrix-facing face of the mitochondrial inner membrane. These findings will allow the distribution of mitochondria-targeted TPP compounds within mitochondria to be fine-tuned.     

Effects of the amount of fillers and of the crosslink density on the mechanical behavior of carbon‐black filled styrene butadiene rubbers

Several carbon‐black filled styrene‐butadiene rubbers are subjected to monotonic uniaxial tension tests in order to investigate the effects of the amount of fillers and of the crosslink density on their mechanical properties. The Young modulus, the volume changes associated with material damage and the stretch to failure are extracted and discussed. Results compare well to the literature results when exist and quantitative analysis are proposed when possible. Results show that filled rubbers are not incompressible when submitted to uniaxial tension tests and their volume changes are strongly dependent of the amount of fillers but are unaffected by the crosslink density. The latter shows strong impact on the filled rubbers stretch to failure but more interestingly this impact is comparable to what is encountered in unfilled rubbers. The stretch to failure is improved by the addition of fillers with an optimum for material filled around 30 phr. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci., 2013     

Novel nucleophilic/basic and acidic organocatalysts for reaction between poorly reactive diisocyanate and diols

Original basic or acidic organic compounds derived from guanidine or phenyl phosphonic acid were specifically designed and tested as new catalysts for the bulk synthesis of polyurethane prepolymers from a precursor system with particularly low reactivity (secondary alcohol + aliphatic isocyanate at low temperature). Both families showed interesting catalytic activities at 60–80 °C, but must nevertheless be used in much higher amounts (1 mol%, i.e. between 0.15 and 0.50 wt%) than traditional metal-based catalysts. The efficiency of guanidine derivatives seems to be related to their nucleophilicity, whereas that of phosphonic acid derivatives depends on their acidity. However, the solubility of the considered species in the reactive medium also plays a major role. The water/alcohol selectivity of the catalysts, especially at room temperature, was then examined as an additional criterion. Guanidines are not selective and favor the reaction of isocyanate groups with water as much as that with alcohols. Phenyl phosphonic acid derivatives are more selective, and particularly pentafluorophenyl phosphonic acid displays a remarkable catalytic activity together with an acceptable selectivity and could represent an interesting and safer alternative to toxic tin and mercury derivatives for many industrial polyurethanes.     

Asymmetric synthesis and biological activity of nor-α-tocopherol, a new vitamin E analogue

The vitamin E analogues (2R,4′R,8′R)‐nor‐α‐tocopherol (94 % de) and (2RS,4′R,8′R)‐nor‐α‐tocopherol have been synthesized from (all R)‐hexahydrofarnesol and phytol, respectively. According to in vitro experiments with murine macrophages nor‐α‐tocopherol is an anti‐inflammatory compound more potent than α‐tocopherol.