Dental caries is a biofilm-related disease, widely perceived to be caused by oral ecological imbalance when cariogenic/aciduric bacteria obtain an ecological advantage. Compared with planktonic bacteria, dental plaques are difficult to remove under extracellular polymeric substance protection. In this study, the effect of caffeic acid phenethyl ester (CAPE) on a preformed cariogenic multi-species biofilm was evaluated, which was comprised of cariogenic bacteria (Streptococcus mutans), commensal bacteria (Streptococcus gordonii), and a pioneer colonizer (Actinomyces naeslundii). Our result revealed that treatment with 0.08 mg/mL CAPE reduced live S. mutans in the preformed multi-species biofilm while not significantly changing the quantification of live S. gordonii. CAPE significantly reduced the production of lactic acid, extracellular polysaccharide, and extracellular DNA and made the biofilm looser. Moreover, CAPE could promote the H2O2 production of S. gordonii and inhibit the expression of SMU.150 encoding mutacin to modulate the interaction among species in biofilms. Overall, our results suggested that CAPE could inhibit the cariogenic properties and change the microbial composition of the multi-species biofilms, indicating its application potential in dental caries prevention and management.

• MeSH
• biofilmy MeSH
• lidé MeSH
• matrix extracelulárních polymerních látek metabolismus   MeSH
• peroxid vodíku * metabolismus   MeSH
• Streptococcus mutans metabolismus   MeSH
• zubní kaz * prevence a kontrola   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

The Arabidopsis EH proteins (AtEH1/Pan1 and AtEH2/Pan1) are components of the endocytic TPLATE complex (TPC) which is essential for endocytosis. Both proteins are homologues of the yeast ARP2/3 complex activator, Pan1p. Here, we show that these proteins are also involved in actin cytoskeleton regulated autophagy. Both AtEH/Pan1 proteins localise to the plasma membrane and autophagosomes. Upon induction of autophagy, AtEH/Pan1 proteins recruit TPC and AP-2 subunits, clathrin, actin and ARP2/3 proteins to autophagosomes. Increased expression of AtEH/Pan1 proteins boosts autophagosome formation, suggesting independent and redundant pathways for actin-mediated autophagy in plants. Moreover, AtEHs/Pan1-regulated autophagosomes associate with ER-PM contact sites (EPCS) where AtEH1/Pan1 interacts with VAP27-1. Knock-down expression of either AtEH1/Pan1 or VAP27-1 makes plants more susceptible to nutrient depleted conditions, indicating that the autophagy pathway is perturbed. In conclusion, we identify the existence of an autophagy-dependent pathway in plants to degrade endocytic components, starting at the EPCS through the interaction among AtEH/Pan1, actin cytoskeleton and the EPCS resident protein VAP27-1.

• MeSH
• aktiny metabolismus   MeSH
• Arabidopsis metabolismus   ultrastruktura   MeSH
• autofagie MeSH
• autofagozomy metabolismus   ultrastruktura   MeSH
• biologické modely MeSH
• buněčná membrána metabolismus   ultrastruktura   MeSH
• endocytóza * MeSH
• endoplazmatické retikulum metabolismus   ultrastruktura   MeSH
• fylogeneze MeSH
• komplex proteinů 2-3 souvisejících s aktinem metabolismus   MeSH
• mikrofilamenta metabolismus   MeSH
• mikrofilamentové proteiny metabolismus   MeSH
• proteiny huseníčku metabolismus   MeSH
• Saccharomyces cerevisiae - proteiny metabolismus   MeSH
• vazba proteinů MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Citrulline ureidase (CTU, EC3.5.1.20) degrades citrulline into ornithine, carbon dioxide, and ammonia. Here, we present the report on expression of recombinant CTU in Escherichia coli. The soluble and active recombinant CTU was expressed in the periplasmic space with the vector pET-22b and the His-tagged CTU was purified with Ni-Affinity Chromatography. The yield of soluble recombinant protein was significantly increased when 1% sorbitol was supplemented in medium. By using phenylisothiocyanate (PITC) pre-column derivatization HPLC, the enzyme activity of recombinant CTU was determined via measuring of the substrate citrulline and the corresponding products. Our results could be useful in the study of CTU biochemical characteristics, enzymatic preparation of ornithine and development of an enzymatic detection method of citrulline.

• MeSH
• Bacteria MeSH
• bakteriologické techniky metody   využití   MeSH
• chromatografie afinitní metody   využití   MeSH
• citrulin * izolace a purifikace   metabolismus   MeSH
• enzymatické testy metody   využití   MeSH
• Escherichia coli enzymologie   imunologie   izolace a purifikace   MeSH
• Francisella tularensis * enzymologie   izolace a purifikace   metabolismus   MeSH
• hydrolasy izolace a purifikace   MeSH
• ornithin * izolace a purifikace   metabolismus   MeSH
• rekombinantní proteiny genetika   izolace a purifikace   metabolismus   MeSH
• sorbitol diagnostické užití   MeSH
• statistika jako téma MeSH
• tularemie diagnóza   etiologie   mikrobiologie   MeSH
• ureasa izolace a purifikace   metabolismus   MeSH
• vysokoúčinná kapalinová chromatografie metody   využití   MeSH
• Publikační typ
• práce podpořená grantem MeSH