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Egg white peptides (EWPs) can be effectively used to alleviate and treat inflammatory diseases due to their anti-oxidation, anti-inflammation, and microbiota regulation capabilities. A dextran sodium sulfate (DSS)-induced colitis model was used to clarify the regulatory effects of EWPs on colitis. Forty-three peptide sequences were identified from EWPs using LC-MS/MS. The results demonstrated that EWPs decreased the levels of pro-inflammatory cytokines and the extent of crypt damage in a dose-dependent manner. 16S rRNA gene sequencing results indicated that 200 mg/kg EWPs significantly increased the relative abundance of beneficial bacteria Lactobacillus and Candidatus Saccharimonas, and reduced the relative abundance of pathogenic bacteria Ruminiclostridium and Akkermansia. In addition, the degree of correlation between pro-inflammatory cytokines and microbiota was as follows: interleukin (IL)-1 beta > IL-8 > IL-6 > tumor necrosis factor-alpha To summarize, EWPs contributed to the alleviation of colitis symptoms and the intestinal injury through anti-inflammatory effects, repair of intestinal mucosa, and modulation of gut microbiota.

De intracellulaire omgeving verschilt van het extracellulaire milieu door de hoge dichtheid in macromoleculen. Deze hoge dichtheid aan macromoleculen, macromoleculaire crowding genoemd, kan worden gekwantificeerd met intracellulaire eiwit-gebaseerde sensoren. De mate van crowding wordt bepaald door Förster resonance energy transfer (FRET) tussen twee verschillende fluorescente eiwitten aan de sensor. In dit proefschrift helder ik het mechanisme van crowding sensing op, waarbij duidelijk wordt dat de sensoren zich gedragen als polymeren en dat hun compressie gerelateerd is aan hun grootte en de crowderconcentratie. In cellen is de compressie eveneens afhankelijk van de structuur van de linkerregio in de sensor, wat een belangrijk verschil vormt tussen in vitro en in vivo experimenten. Ik heb een afwijking geïdentificeerd vanwege de langzame maturatie van fluorescente eiwitten en dit opgelost door de expressie van de sensor onder een constitutieve promotor te plaatsen. Door crowding te volgen in Escherichia coli hebben we ontdekt hoe crowding verandert in cellen. Eveneens hebben we de eerste sensoren ontwikkeld om de ionische sterkte in levende cellen te bepalen. Deze sensoren om crowding en ionische sterkte te bepalen stellen ons in staat de fysicochemische aspecten in de cel te begrijpen en dienen als eerste stap om de rol van fysisch-chemische parameters op de biochemische organisatie en fysiologie van de levende cel op te helderen., The high density of macromolecules makes the intracellular environment markedly different from bulk solution. It was recently shown that the high density, also described as macromolecular crowding, can be quantified with protein-based sensors inside cells. These sensors are based on Förster resonance energy transfer (FRET). In this thesis, I reveal the crowding sensing mechanism, showing that the sensors behave as polymers and their compression scales with their size and crowder concentration. In cells, compression also depends on the structure of the sensor linker region, which shows an important difference between experiments in buffer versus in the cell. I identify an artifact caused by slow maturation of the fluorescent proteins, and resolve this by sensor expression under a constitutive promoter. By tracking the crowding changes in Escherichia coli, we reveal how crowding changes in cells. Finally, we developed the first sensors to determine the ionic strength in living cells. These sensors for the crowding and ionic strength will allow us to deepen our understanding of the physicochemical aspects in the cell and serve as a stepping-stone to elucidate the role of physical chemical parameters on the biochemical organization and well-being of the living cell.