• Title/Summary/Keyword: HPLC analysis

Search Result 2,671, Processing Time 0.03 seconds

Preparation of Selenium-enriched Bifidobacterium Longum and its Effect on Tumor Growth and Immune Function of Tumor-Bearing Mice

  • Yin, Yan;Wang, Rong-Rong;Wang, Yan;Wang, Jian-Jun;Xu, Gen-Xing
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.15 no.8
    • /
    • pp.3681-3686
    • /
    • 2014
  • In this study, we demonstrated selenium (Se) accumulation in Bifidobacterium longum strain (B. longum) and evaluated the effect of Se-enriched B. longum (Se-B. longum) on tumor growth and immune function in tumor-bearing mice. Analysis using high-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP-MS) revealed that more than 99% of Se in Se-B. longum was organic, the main component of which was selenomethionine (SeMet). In the in vivo experiments, tumor-bearing mice (n=8) were orally administrated with different doses of Se-B. longum alone or combined with cyclophosphamide (CTX). The results showed that the middle and high dose of Se-B. longum significantly inhibited tumor growth. When Se-B. longum and CTX were combined, the antitumor effect was significantly enhanced and the survival time of tumor-bearing mice (n=12) was prolonged. Furthermore, compared with CTX alone, the combination of Se-B. longum and CTX stimulated the activity of natural killer (NK) cells and T lymphocytes, increasing the levels of interleukin-2 (IL-2) and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and the leukocyte count of H22 tumor-bearing mice (n=12).

Identification of Essential Histidines in Cyclodextrin Glycosyltransferase Isoform 1 from Paenibacillus sp. A11

  • Kaulpiboon, Jarunee;Pongsawasdi, Piamsook
    • BMB Reports
    • /
    • v.36 no.4
    • /
    • pp.409-416
    • /
    • 2003
  • The isoform 1 of cyclodextrin glycosyltransferase (CGTase, EC 2.4.1.19) from Paenibacillus sp. A11 was purified by a preparative gel electrophoresis. The importance of histidine, tryptophan, tyrosine, and carboxylic amino acids for isoform 1 activity is suggested by the modification of the isoform 1 with various group-specific reagents. Activity loss, when incubated with diethylpyrocarbonate (DEP), a histidine modifying reagent, could be protected by adding 25 mM methyl-$\beta$-cyclodextrin substrate prior to the modification. Inactivation kinetics of isoform 1 with DEP resulted in second-order rate constants ($k_{inactivation}$) of $29.5\;M^{-1}s^{-1}$. The specificity of the DEP-modified reaction for the histidine residue was shown by the correlation between the loss of isoform activity and the increase in the absorbance at 246 nm of N-carbethoxyhistidine. The number of histidines that were modified by DEP in the absence and presence of a protective substrate was estimated from the increase in the absorbance using a specific extinction coefficient of N-carbethoxyhistidine of $3,200\;M^{-1}cm^{-1}$. It was discovered that methyl-$\beta$-CD protected per mole of isoform 1, two histidine residues from the modification by DEP. To localize essential histidines, the native, the DEP-modified, and the protected forms of isoform 1 were digested by trypsin. The resulting peptides were separated by HPLC. The peptides of interest were those with $R_t$ 11.34 and 40.93 min. The molecular masses of the two peptides were 5,732 and 2,540 daltons, respectively. When the data from the peptide analysis were checked with the sequence of CGTase, then His-140 and His-327 were identified as essential histidines in the active site of isoform 1.

Breast Cancer Association with CYP1A2 Activity and Gene Polymorphisms - a Preliminary Case-control Study in Tunisia

  • Ayari, I;Arnaud, MJ;Mani, A;Pavanello, S;Saguem, S
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.16 no.8
    • /
    • pp.3559-3563
    • /
    • 2015
  • The aim of the present study was to evaluate the relative contribution of CYP1A2 isoforms (-3860 G/A, -2467T/delT and -163C/A) in control subjects and breast cancer patients to the metabolism of caffeine in human liver. Restriction fragment length polymorphism analysis of PCR-amplified Fragments (PCR-RFLP) was used for the genotyping of CYP1A2 SNPs and HPLC allowed the phenotyping through the measurement of CYP1A2 activity using the 17X + 13X + 37X/137X urinary metabolite ratio (CMR) and plasma caffeine half life (T1/2). The CYP1A2 -3860A genotype was associated with a decreased risk of breast cancer. In contrast, distributions of the CYP1A2 -2467T/delT or -2467delT/delT and -163A/C or A/A genotypes among breast cancer patients and controls were similar. When the genotype and phenotype relationship was measured by comparing the mean CMR ratios and caffeine half life within the genotype groups between subjects and breast cancer patients, there were no significant differences except for -3860 A, most of them being homozygous for the -3860 G/G SNP and had a significant higher mean CMR ratio and half life than those with -3860 G/A (P=0.02). The results of this preliminary study show a significant association between CP1A2 -3860 G variant and CYP1A2 phenotype which must be confirmed by further large-size case-control studies.

Phytoplankton Distribution in the Eastern Part of the Yellow Sea by the Formation of Tidal Front and Upwelling during Summer (황해 동부 해역에서 하계에 조석전선과 용승에 의한 식물플랑크톤군집 분포)

  • Lee, Young-Ju;Choi, Joong-Ki;Shon, Jae-Kyoung
    • Ocean and Polar Research
    • /
    • v.34 no.2
    • /
    • pp.111-123
    • /
    • 2012
  • To understand the phytoplankton community in the eastern part of the Yellow Sea (EYS), in the summer, field survey was conducted at 25 stations in June 2009, and water samples were analyzed using a epifluorescence microscopy, flow cytometry and HPLC method. The EYS could be divided into four areas by a cluster analysis, using phytoplankton group abundances: coastal mixing area, Anma-do area, transition water, and the central Yellow Sea. In the coastal mixing area, water column was well mixed vertically, and phytoplankton was dominated by diatoms, chrysophytes, dinoflagellates and nanoflagellates, showing high abundance ($>10^5\;cells\;l^{-1}$). In Anma-do coastal waters characterized by high dominance of dinoflagellates, high phytoplankton abundance and biomass separated from other coastal mixing area. The southeastern upwelling area was expanded from Jin-do to Heuksan-do, by a tidal mixing and coastal upwelling in the southern area of Manjae-do, and phytoplankton was dominated by benthic diatoms, nanoflagellates and Synechococcus group in this area. Phytoplankton abundance and biomass dominated by pico- and nanophytoplankton were low values in the transition waters and the central Yellow Sea. In the surface of the central Yellow Sea, high dominance of photosynthetic pigments, 19'-hexanoyloxyfucoxanthin and zeaxanthin implies that haptophytes and cyanobacteria could be the dominant group during the summer. These results indicate that the phytoplankton communities in the EYS were significantly affected by the formation of tidal front, thermal stratification, and coastal upwelling showing the differences of physical and chemical characteristics during the summer.

The Relationship between Virginiae Butanolide C(VB-C) and Receptor in Virginiamycin Production (Virginiamycin 생산유도에 관여하는 Virginiae Butanolide C(VB-C) 및 Receptor의 상관관계)

  • Kim, Hyun-Soo;Hyun, Ji-Sook;Yu, Tae-Shick
    • Microbiology and Biotechnology Letters
    • /
    • v.24 no.1
    • /
    • pp.59-66
    • /
    • 1996
  • Virginiae butanolide C(VB-C) is one of the butyrolactone autoregulators, which triggers the productin of virginiamycin in Streptomyces virginiae. To further understand the mechanism of virginiamycin induction, we isolated three mutants from S. virginiae by N-methyl-N'-nitrosoguanidine (NTG) treatment. The characteristics of the three mutants were confirmed as follows: the mutant No. 1 delayed the production of the VB-C, receptor and antibiotics; the mutant No.3 hyperproduced receptor; the mutant No.4 failed to produce the VB-C. The addition of synthetic VB-C couldn't induce the production of antibiotics in the mutant No.1 due to delayed production of receptor, could provoke the production of larger amount of antibiotics than parental wild type strain in the mutant No.3 due to the presence of large amount of receptor, and could induce production of very small amount of antibiotics in the mutant No.4 due to the absence of VB-C. Antimicrobial spectrum and HPLC analysis of the mutant No.1 and No.3 suggested that the VB-C might have a specific ability to induce the production of virginiamycin M and S. These results imply that the VB-C has an ability to trigger the production of virginiamycin under receptor existence in S. virginiae.

  • PDF

Vitamin A Nutritional Status Determined by Assessing Dietary Intake and Serum Retinol Level among Elderly Adults Living in Asan (아산시 거주 노인들의 비타민 A 섭취 실태 및 레티놀 영양상태)

  • Kwon, Young-Suk;Kim, Hee-Seon
    • Korean Journal of Human Ecology
    • /
    • v.13 no.1
    • /
    • pp.75-84
    • /
    • 2004
  • Insufficient dietary intake of vitamin A is one of the major nutritional problems for elderly adults in some parts of Korea. The objective of this study was to determine the vitamin A nutritional status of elderly adults in Asan, Korea by assessing the dietary intake and serum retinol concentration. Five hundred twenty four subjects (218 male and 306 female) over 65 years were recruited from city of Asan. Each subject was interviewed to assess the intake of vitamin A using a 24hr recall method and data were analysed from computer-aided nutrient analysis program. Blood samples after 12hr fasting were collected for serum retinol concentration and reverse phased HPLC with UV detector used. The results showed that subjects did not consume the sufficient amount of energy (82-85% of Korean RDA for male and 77-79% RDA for female) and vitamin A (59% RDA for male and 50% RDA for female). Range for retinol intake was 0 to $4342\;{\mu}g$ a day while that of beta-carotene was 65 to $31595\;{\mu}g$. Serum retinol concentrations were within a normal range for both male ($80\;{\mu}g/dl$) and female ($67\;{\mu}g/dl$) subjects. Many subjects (n=342) consumed less than 50% RDA of vitamin A. However, if retinol intake was high (> $37\;{\mu}g$), even with less than 50% RDA of vitamin A intake, serum retinol concentration was high ($75\;{\mu}g/dl$). Subjects showed normal serum retinol status even with low vitamin A intake. The results suggested that optimal intake ratio of dietary retinol and carotenoid is important to maintain an appropriate serum retinol concentration.

  • PDF

Separation and Characteristics of ADH and ALDH Activators in Fermented Lycii fructus Extract (발효 전후 구기자열매 추출물의 ADH 및 ALDH 활성인자 분리 및 특성)

  • Choi, Ji-Hyun;Kang, Dong-Hee;Kim, Hyun-Soo
    • KSBB Journal
    • /
    • v.31 no.2
    • /
    • pp.134-143
    • /
    • 2016
  • Lycii fructus has been traditionally used as a preventive and therapeutic medicine to treat enervation and diverse chronic diseases. In this study, we investigated whether fermentation of Lycii fructus extract (LE) increases the enzymatic activity of the alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). The fermentation of LE by Bacillus subtilis subsp. subtilis and Saccharomyces cerevisiae IFO 2376 was shown to increase the enzymatic activity of ADH and ALDH. TLC analysis of LE and fermented LE (FLE) showed that ADH and ALDH activities increased in different spots. Fraction No. 66 of LE and fraction No. 68 of FLE by Silica gel chromatography showed increased ADH activity of 129.1% and 148.9%, respectively. Fractions No. 128 of LE and FLE by Silica gel chromatography showed increased ALDH activity of 134.1% and 148.1%, respectively. The fraction No. 68 of FLE obtained by HPLC showed new peaks at $R_t$ 11.938min, $R_t$ 22.072min and $R_t$ 28.842min, indicating that ADH activity was increased. The LE and FLE fractions with the greatest increases in ADH activity peaked at the same time ($R_t$ 13min),whereas the LE and FLE fractions with the greatest increases in ALDH activity peaked at different times ($R_t$ 16.307min and $R_t$ 36.640min, respectively).

Changes of Methyl trans-cinnamate Levels During Fruit-body Development in Tricholoma matsutake

  • Lee, Wi Young;Park, Youngki;Ahn, Jin Kwon;Ka, Kang Hyeon
    • Journal of Korean Society of Forest Science
    • /
    • v.94 no.5 s.162
    • /
    • pp.330-333
    • /
    • 2005
  • The relationship between the contents of methyl trans-cinnamate and the ratio of DP/DS (diameter of pileus / diameter of stalk) in the fruit-body of Tricholoma matsutake during its development was investigated. The stages of development were divided as follows: stage A is less than 1, stage B is from 1 to less than 2, stage C is from 2 to less than 3, and stage D is more than 3 of the values of DP/DS. The contents of methyl trans-cinnamate in pileus and stalk of pine mushroom ranged from $77{\mu}g/g$ to $824{\mu}g/g$ and from $7.6{\mu}g/g$ to $22.4{\mu}g/g$, respectively during its development. In the part of pileus, there is no relevance of the methyl trans-cinnamate content of pine mushroom between the stage A and B, but there was significantly different among the stage of B, C and D. In the case of stalk, the relevance of the methyl trans-cinnamate content of pine mushroom between stage D and other stages showed a low difference. In addition, as pileus of pine mushroom developed the level of the aroma compound increased as well and showed higher correlation relationship ($r^2=0.877$) between the contents of methyl trans-cinnamate in the pileus and the ratio of DP/DS. From the results of this study, we can conclude that the aromatic component of pine mushroom can be deduced from the value of DP/DS, which indicates the stage of the development appearance.

A Study on Vitamin C Content of Nutrition Emphasized Products (영양강조표시제품 중 비타민 C 함량 조사)

  • Jeong, Da-un;Lee, Heon-Ok;Kim, Young-Kyoung;Om, Ae-Son
    • Korean Journal of Community Nutrition
    • /
    • v.21 no.6
    • /
    • pp.574-579
    • /
    • 2016
  • Objectives: Vitamin C has various functions such as antioxidative effect and supporting absorption of iron (Fe). Aim of this present study was to provide vitamin C nutrition information and to briefly evaluate absorption interaction of vitamin C and Fe content of vitamin C emphasized products. Methods: Vitamin C emphasized foods including beverages, cereal, snacks, chocolate products, other cocoa products, and sugary products were examined by HPLC. Fe contents in samples after dry-ashing were examined by ICP. Results: Vitamin C content ranges in various products tested were the following: beverages (n=11) $20.15{\pm}0.08{\sim}845.41{\pm}6.07mg$, cereal (n=11) $52.50{\pm}0.23{\sim}262.50{\pm}0.07mg$, snacks (n=1) $50.00{\pm}0.25mg$, chocolate products (n=1) $311.73{\pm}2.44mg$, other cocoa products (n=1) $311.73{\pm}2.44mg$, other sugary products (n=2) $52.50{\pm}0.23{\sim}262.50{\pm}0.07mg$. Vitamin C (n=27) analysis values ranged from 82 to 450% of the labeled value. Vitamin C content in vitamin C emphasized food (n=6) was estimated 7.7 times~56.6 times more than Fe content. Conclusions: Analyzed samples ranged more than 80% of the labeled value in vitamin C emphasized products, which complied with food labeling regulation. But, beverages (n=3), cereal (n=4), chocolate products (n=1) were 2 times more than the labeled value. To provide accurate nutrition information, food manufactures should supervise nutrition labeling and understand the interactions between nutrients. Also, consumer should decide about the adequate amount of nutrient intake by thoroughly checking nutrition labeling.

Production and Characterization of Monoclonal and Recombinant Antibodies Against Antimicrobial Sulfamethazine

  • Yang, Zheng-You;Shim, Won-Bo;Kim, Min-Gon;Lee, Kyu-Ho;Kim, Keun-Sung;Kim, Kwang-Yup;Kim, Cheol-Ho;Ha, Sang-Do;Chung, Duck-Hwa
    • Journal of Microbiology and Biotechnology
    • /
    • v.17 no.4
    • /
    • pp.571-578
    • /
    • 2007
  • A monoclonal antibody (mab) against the antimicrobial sulfamethazine was prepared and characterized by an indirect competitive enzyme-linked immunosorbent assay (IC-ELISA). Sulfamethazine in the range of 0.2 and 45ng/ml could be determined with the mab by IC-ELISA. cDNAs encoding a variable heavy chain and variable light chain of the mab were cloned to produce recombinant antibodies using phage display technology. Following phage rescue and three rounds of panning, a single-chain variable fragment (scFv) antibody with high sulfamethazine-binding affinity was obtained. ELISA analysis revealed that scFv antibody and parent mab showed similar, but not identical, characteristics. The $IC_{50}$ value by IC-ELISA with scFv antibody was 4.8ng/ml, compared with 1.6ng/ml with the parent mab. Performances of the assays in the presence of milk matrix were compared; the mab-based assay was less affected than the scFv-based assay. Sixty milk samples were analyzed by mab-based IC-ELISA, and four samples were sulfamethazine positive; these results were favorably correlated with those obtained by HPLC.