Enzymatic synthesis of phenolic acid glucosyl esters to test activities on cholangiocarcinoma cells.

While glycoside hydrolase family 1 (GH1) enzymes mostly catalyze hydrolysis reactions, rice Os9BGlu31 preferentially catalyzes transglycosylation to transfer a glucosyl moiety to another aglycone moiety to form a new glycosylated compound through a retaining mechanism. In this study, Os9BGlu31 was used to synthesize eight phenolic acid glucosyl esters, which were evaluated for activities in cholangiocarcinoma cells. The transglycosylation products of Os9BGlu31 wild type and its mutant variants were detected, produced on a milligram scale, and purified, and their structures were characterized by NMR spectroscopy. The transglycosylation products were evaluated by antioxidant and anti-proliferative assays, followed by an anti-migration assay for the selected phenolic acid glucosyl ester. Os9BGlu31 mutants produced higher yield and activity than wild-type enzymes on phenolic acids to produce phenolic acid glucosyl esters. Among these, gallic acid glucosyl ester (ß-glucogallin) had the highest antioxidant activity and anti-proliferative activity in cholangiocarcinoma cells. It also inhibited the migration of cholangiocarcinoma cells. Our study demonstrated that rice Os9BGlu31 transglucosidase is a promising enzyme for glycosylation of bioactive compounds in one-step reactions and provides evidence that ß-glucogallin inhibits cell proliferation and migration of cholangiocarcinoma cells. KEY POINTS: • Os9BGlu31 transglucosidases produced phenolic acid glucosyl esters for bioactivity testing. • Phenolic acid glucosyl esters were tested for cytotoxicity in cholangiocarcinoma cells. • ß-Glucogallin displayed the highest inhibition of cholangiocarcinoma cell growth.

Semi-synthesis of phenolic-amides and their cytotoxicity against THP-1, HeLa, HepG2 and MCF-7 cell lines.

In the present study, we derivatized several hydroxycinnamic and hydroxybenzoic acids to phenolic amides (PAMs) via one step BOP mediated amide coupling reactions. Fifteen PAMs were synthesized in >40% yields and were screened for their cytotoxic activities against four cancer cell lines: THP-1 (leukaemia), HeLa (cervical), HepG2 (liver), and MCF-7 (breast), in comparison to 5-flurouracil (5-FU). Four amides showed IC50 ranging from 5 to 55 µM against all four cell lines. In contrast, tetradecyl-gallic-amide (13) affected only THP-1 leukaemia cells with IC50 of 3.08 µM. The activities of these compounds support the promise of phenolic amides as anticancer agents.

Diversity of Sub-Gingival Fluids Microbiota Compositions in Periodontitis and Rheumatoid Arthritis Patients: A Case-Control Study

Abstract Objective: To determine the diversity of microbiota in the sub-gingival fluids from periodontitis and rheumatoid arthritis (RA) patients and to know the relationship of microbiota abnormalities between periodontitis and RA. Material and Methods: Samples were collected from sub-gingival fluids of nine participants by swabbing technique. The participants were divided into 3 groups, namely the control (not periodontitis and RA), periodontitis, and RA. The total number of bacteria was calculated by using direct culture in serial dilution by applying the total plate count method. Isolation of microbiota was conducted by pour plates and incubated in an anaerobic jar. The bacterial diversity was performed by Simpson index and DNA Isolation. Results: There were differences in the concentration of sub-gingival fluids bacteria in the control group with the periodontitis and RA group. The microbiota composition profiles were different for each group. The bacterial isolate 4, 5 and 22 were found in all groups, while isolate 14 was assumed to be related to the metabolic pathway in microbiota abnormalities associated with disease progression. Conclusion: The Simpson index has a value >0.61 with high bacterial dominance; however, the diversity of microbiota is at a low level of diversity. We assumed that isolate 14 was associated with development of RA disease.