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Untitled Prezi

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Alessandro Prestes

on 6 August 2013

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During digestion (with pancreatin and pepsin)
Results indicate:
In the cells (HTC116, MOLT-4 and HeLLaS3):
Cytotoxicity of alpha-dicarbonyl compounds submitted to in vitro simulated digestion process
Materials and Methods
Alessandra Amoroso, Giovanni Maga, Maria Daglia
On DNA enzymes (DNA pol beta and DNA pol lambida):
Institute of Molecular Genetics, IGM-CNR, via Abbiategrasso 207, 27100 Pavia, Italy

Department of Drug Sciences, University of Pavia, via Taramelli 12, 27100 Pavia, Italy

Food Chemistry 140 (2013) 654–659
Impact Factor: 3.334
In this study the interactions between a mixture of glyoxal, methylglyoxal and 2,3-butanedione and the digestive enzymes (pepsin and pancreatin) were studied. The results showed that during gastroduodenal digestion a-DCs react with digestive enzymes to produce carbonylated proteins. Moreover, undigested and digested a-DC cytotoxicity against human cells, as well as their ability to inhibit the function of human enzymes responsible for DNA repair were shown.
a-Dicarbonyl compounds (a-DCs), such as glyoxal, methylglyoxal and 2,3-butanedione, are highly reactive substances occurring in thermally treated and fermented foods, that may react with amino and sulphydryl groups of side chains of proteins to form Maillard reaction end products, inducing a negative impact on the digestibility and on nutritional value of protein. In recent years the role of food derived a-DCs in gastroduodenal tract is under investigation to understand whether excess consumption of such dietary compounds might be a risk for human health.
glyoxal (GO), methylglyoxal (MGO), 2,3-butanedione (2,3-BD)
submitted to in vitro simulated gastroduodenal
digestion (following the method described by Ames et al. (1999) with some modifications).
after gastroduodenal digestion: RP-HPLC-DAD method evaluated the concentration of GO, MGO and 2,3-BD
RP-HPLC-DAD method validation
Quantification of GO, MGO, and 2,3-BD was performed following ICH procedures (2012).


limit of detection
Determination of digestive enzyme (pepsin and pancreatin) carbonylation by GO, MGO and 2,3-BD
determined by derivatising the protein carbonyl adducts with 2,4-dinitrophenylhidrazine (DNPH) as reported by Dong, Banaich, and O’Brien (2010)
Enzymatic assays
Human DNA polymerase beta and lambda activity
Cell culture and viability assays
MOLT-4 (a human early T cell leukaemia cell line), HeLaS3 (a human epithelial carcinoma cell line) and HCT116 (a human colon cancer cell line) were used.
alpha-DCs react with digestive enzymes to produce carbonylated proteins
undigested and digested alpha-DC cytotoxicity was observed (decreases the cell viability)
inhibit the function of human enzymes responsible for DNA repair
A total of 18 a-DCs has been identified so far from different food matrices,
2,3-butanedione (2,3-BD)
Alpha-dicarbonyl compounds (naturally in foods)
Alpha-DC after gastroduodenal digestion??
HPLC to quantification of MG, GO and 2,3-BD
After digestion, MG, GO and 2,3-BD are present in medium
Measurements to method validation
Alpha-DC increase the protein carbonilation
increase in EC50
increase in EC50
increase in EC50
decrease in EC50
decrease in EC50
decrease in EC50
less resistance
high resistance
intermediate resistance
Full transcript