Carbohydrate-protein interactions investigated on plastic chips statically coated with hydrophobically modified hydroxyethylcellulose.

We developed a novel method for rapid screening of carbohydrate-protein interactions using poly(methyl methacrylate) (PMMA) channels statically coated with hydrophobically modified hydroxyethylcellulose (HM-HEC). We found that a self-assembled monolayer (SAM) of HM-HEC on a PMMA surface intact by water allows rapid and reproducible separations of glycan samples using a 20 mM phosphate without HM-HEC. The underlying mechanism for dynamic and static coatings on the PMMA surface is discussed. Simultaneous analysis of the molecular interaction between a complex mixture of carbohydrates from alpha1-acid glycoprotein and proteins has been successfully achieved in PMMA channels statically coated with a SAM of HM-HEC.

No association between Helicobacter pylori seropositivity and ornithine decarboxylase (ODC) A317G polymorphism, and no modification by NAD(P)H:qinone oxidoreductase 1 (NQO1) C609T.

Our previous epidemiologic study reported that NAD(P)H:quinone oxidoreductase 1 (NQO1) 609C/C with full enzyme activity was a high risk genotype for Helicobacter pylori (H. pylori) seropositivity. Since NQO1 stabilizes ornithine decarboxylase (ODC), which attenuates the innate immune response through elevated polyamines, ODC functional polymorphisms may also influence H. pylori seropositivity. This study aimed to examine the association with ODC A317G polymorphism, as well as the modification by NQO1 C609T. The two polymorphisms were determined by polymerase chain reaction with confronting two-pair primers (PCR-CTPP) among 465 health checkup examinees in Nagoya. The ODC A317G genotype frequency was 35.9% for A/A, 49.3% for A/G, and 14.8% for G/G. The sex-age-adjusted odds ratio (OR) of the ODC gene for H. pylori seropositivity was not significant (OR = 1.09 for G/A and OR = 1.02 for G/G, relative to A/A). Among subjects with any NQO1 genotype, no association was observed between the ODC ploymorphism and H. pylori seropositivity. Results of the present study did not support the hypothesis that the different genetic traits in the ODC-polyamine pathway are associated with susceptibility to persistent H. pylori infection. The higher frequency of the ODC 317A allele in the Japanese population than that in the Caucasian population is firstly reported. The genetic traits through the ODC-polyamine pathway will be further investigated.

Eradication rate of Helicobacter pylori according to genotypes of CYP2C19, IL-1B, and TNF-A.

BACKGROUND: Lansoprazole, amoxicillin, and clarithromycin are commonly used drugs for eradication of Helicobacter pylori (H. pylori). A few studies reported that the eradication rate was influenced by the functional polymorphism of CYP2C19, whose product metabolizes proton pomp inhibitors including lansoprazole. METHODS: This study examined the eradication rate among 67 participants in the polymorphism study who visited Daiko Medical Center, Nagoya University from July 2004 to October 2005. The participants aged 20 to 69 years were classified into three group according to CYP2C19 genotype; rapid metabolizers (RM) with *1*1 genotype, intermediate metabolizers (IM) with *1*2 or *1*3 genotype, and poor metabolizers (PM) with *2*2, *2*3, or *3*3 genotype. For the genotype classification, G681A (681G for *1 and 681A for *2) and G636A (636G for *1 and 636A for *3) were genotyped by PCR with confronting two-pair primers (PCR-CTPP). They were also genotyped for IL-1B T-31C and TNF-A T-1031C by a duplex PCR-CTPP. RESULTS: The eradication rate was 70.0% for RM, 93.9% for IM, and 85.7% for PM. The difference in the rate between RM and IM+PM was statistically significant (p=0.025). The eradication rate was highest for those with IL-1B -31CC; the p value was marginal among the whole subjects (chi2=3.78, p=0.05) and not significant among the RM group (chi2=1.60, p=0.21). The genotypes of TNF-A T-1031C had no associations with the eradication rate. But among the RM group, the odd ratio (OR) of the TNF-A CT for the eradication rate relative to TT was marginally reduced (OR=0.05, 95% confidence interval, 0.002-1.19). CONCLUSIONS: The present study confirmed the low eradication rate for RM. The reproduced finding provides evidence that the CYP2C19 genotype is useful to predict the success of the treatment. For the RM group, alternative regimens expected to be with a higher eradication rate will be recommended, especially to those with the TNF-A -1031C allele.

Influence of the pH on separating DNA by high-speed microchip electrophoresis.

Various factors are critical in resolving DNA molecules at high speed, including the separation medium, concentration, composition, and pH of the buffer, as well as the electric field strength. To this study, considered the composition of a buffer and the difference in the pH, while paying attention to whether the separation ability changes in the microchip electrophoresis of DNA. DNA separation was particularly affected by both the buffer composition and the pH. Under the optimal microchip electrophoresis conditions that were determined in this study, an improved resolution of a wider range of DNA fragment sizes was achieved. Moreover, the total separation time decreased from 240 s to 100 s. Thus, by simplifying and improving the DNA electrophoresis in the microchip, this technique is now widely applicable to several different scientific fields.