ABSTRACT
Forty-four female students with a tendency for constipation (mean age, 20.2±3.3 y) were asked to consume 30 g test pickles daily for 2 wk and were divided into 3 groups: viable-cell intake subjects (n=14, 3.0×105 colony-forming units of viable LAB (lactic acid bacteria) cells per sample), dead-cells intake subjects (n=15, viable cells were heat sterilized), and placebo-intake subjects (n=15, LAB removed from the pickles). γ-Aminobutyric acid content of 75.1±3.2 mg per sample was noted, with no marked difference between samples containing viable and dead cells. Natural killer (NK)-cell activity (% specific lysis) in serum from dead-cell intake subjects was 37.5±17.0% before the start of the test-food intake and 47.7±20.1% after intake, indicating statistically significant effects (p<0.01). However, viable-cell intake and placebo intake subjects showed no statistically significant difference. The number of days with bowel movements significantly increased from 3.8±1.5 to 4.9±1.8 d in the dead-cell intake group, whereas a slight change from 4.6±1.5 to 5.1±1.7 d was observed in the viable-cell intake group. Additionally, the feeling of incomplete evacuation fell and a refreshed feeling increased among the subjects with constipation. Thus, marked enhancement of NK-cell activity and improved bowel symptoms were observed in subjects consuming pickles containing dead LAB cells.
Subject(s)
Constipation/diet therapy , Food, Preserved/microbiology , Functional Food/microbiology , Immunomodulation , Intestines/physiopathology , Levilactobacillus brevis/growth & development , Vegetables/microbiology , Adult , Constipation/immunology , Constipation/microbiology , Constipation/physiopathology , Diet/ethnology , Feces/chemistry , Female , Humans , Intestines/immunology , Intestines/microbiology , Japan , Killer Cells, Natural/immunology , Levilactobacillus brevis/immunology , Levilactobacillus brevis/isolation & purification , Levilactobacillus brevis/metabolism , Microbial Viability , Odorants , Severity of Illness Index , Sterilization , Young Adult , gamma-Aminobutyric Acid/analysisABSTRACT
Owing to the increasing importance of genomic information, obtaining genomic DNA easily from biological specimens has become more and more important. This article proposes an efficient method for obtaining genomic DNA from nail clippings. Nail clippings can be easily obtained, are thermostable and easy to transport, and have low infectivity. The drawback of their use, however, has been the difficulty of extracting genomic material from them. We have overcome this obstacle using the protease solution obtained from Cucumis melo. The keratinolytic activity of the protease solution was 1.78-fold higher than that of proteinase K, which is commonly used to degrade keratin. With the protease solution, three times more DNA was extracted than when proteinase K was used. In order to verify the integrity of the extracted DNA, genotype analysis on 170 subjects was performed by both PCR-RFLP and Real Time PCR. The results of the genotyping showed that the extracted DNA was suitable for genotyping analysis. In conclusion, we have developed an efficient extraction method for using nail clippings as a genome source and a research tool in molecular epidemiology, medical diagnostics, and forensic science.
Subject(s)
Cucumis melo/enzymology , DNA/isolation & purification , Molecular Biology/methods , Nails/chemistry , Peptide Hydrolases/metabolism , Alleles , Deoxyribonucleases/antagonists & inhibitors , Edetic Acid/pharmacology , Endopeptidase K/metabolism , Female , Fruit/enzymology , Genome, Human/genetics , Humans , Hydrogen-Ion Concentration/drug effects , Keratins/metabolism , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Solutions , Temperature , Young AdultABSTRACT
This is the first report of glycoside hydrolase family 43 beta-xylosidase from Aspergillus oryzae. To characterize this enzyme, the recombinant enzyme was expressed in Escherichia coli. Unlike known beta-xylosidases from fungal origins, the enzyme did not show substrate ambiguity and was stable at alkaline pH.
