• Title/Summary/Keyword: metabolites analysis

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The Gut Microbial Lipid Metabolite 14(15)-EpETE Inhibits Substance P Release by Targeting GCG/PKA Signaling to Relieve Cisplatin-Induced Nausea and Vomiting in Rats

  • Man Lu;Liwei Xie;Sijie Yin;Jing Zhou;Lingmei Yi;Ling Ye
    • Journal of Microbiology and Biotechnology
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    • v.34 no.9
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    • pp.1769-1777
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    • 2024
  • Chemotherapy-induced nausea and vomiting (CINV) is a debilitating side effect related to activation of substance P (SP). SP activation can result from dysregulation of the gut-brain axis, and also from activation of protein kinase A signaling (PKA) signaling. In this study, we connected these factors in an attempt to unveil the mechanisms underlying CINV and develop new therapeutic strategies. Female rats were injected with cisplatin (Cis) to induce pica. Fecal samples were collected before/after injection, and subjected to lipid metabolomics analysis. In another portion of pica rats, the PKA inhibitor KT5720 was applied to investigate the involvement of PKA signaling in CINV, while fecal microbiota transplantation (FMT) was implemented to verify the therapeutic effect of the lipid metabolite 14(15)-EpETE. Pica symptoms were recorded, followed by ileal histological examination. The targeting relationship between 14(15)-EpETE and glucagon was determined by bioinformatics. SP and glucagon/PKA signaling in rat ileum, serum, and/or brain substantia nigra were detected by immunohistochemistry, enzyme-linked immunosorbent assay, and/or western blot. The results showed a significantly lower level of 14(15)-EpETE in rat feces after Cis injection. KT5720 treatment alleviated Cis-induced pica symptoms, ileal injury, SP content increase in the ileum, serum, and brain substantia nigra, and ileal PKA activation in rats. The ileal level of glucagon was elevated by Cis in rats. FMT exerted an effect similar to that of KT5720 treatment, relieving the Cis-induced changes, including ileal glucagon/PKA activation in rats. Our findings demonstrate that FMT restores 14(15)-EpETE production, which inhibits SP release by targeting GCG/PKA signaling, ultimately mitigating CINV.

Monitoring of Natural Preservative Levels in Food Products (가공식품 중 천연유래 보존료 함량에 대한 조사)

  • Park, Eun-Ryong;Lee, Sun-Kyu;Hwang, Hye-Shin;Mun, Chun-Sun;Gwak, In-Shin;Kim, Ok-Hee;Lee, Kwang-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.12
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    • pp.1640-1646
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    • 2008
  • In the current food sanitation regulation, food additives are under controlled by the Food Code. The naturally derived preservatives such as benzoic acid and propionic acid can be naturally carried over or produced as metabolites during manufacturing process such as fermentation. To monitor naturally formed benzoic acid and propionic acid levels, a total of 145 samples were classified into berries (prune, cranberry), functional foods (propolis liquid, ginseng product), vinegars (vinegar-based drink, vinegar beverage, vinegar), and salted and pickled products (olive, pickled cucumber, salted/pickled product) and analyzed by HPLC-PDA and GC-FID. From the results, benzoic acid and propionic acid were each detected and identified in 144 samples and 64 samples respectively. The amount of benzoic acid ranged from $4.1{\sim}478.4\;ppm$ in cranberry, from $49.7{\sim}491$ in propolis liquid, and from $2.5{\sim}10.2\;ppm$ in ginseng, and other tested samples contained very small quantity. Also, the amount of propionic acid ranged from $179.8{\sim}951.9\;ppm$ (av. 553.6 ppm) in vinegar (persimmon vinegar 100%), which was the highest level among fermented foods, from $13.7{\sim}247.0$ ppm in propolis liquid, from $2.0{\sim}180.7\;ppm$ in vinegar-based drink, and from $1.6{\sim}76.6\;ppm$ in olive. Vinegar beverage and pickled cucumber each showed 24 and 18 ppm of propionic acid; in contrast, propionic acid was not detected in prune, cranberry, ginseng, and picked/salted products.

Analysis of ethyl glucuronide (EtG) in Hair for the diagnosis of chronic alcohol abuse of Korean (한국인의 만성 알코올 중독 진단을 위한 모발에서 Ethyl Glucuronide (EtG) 분석법 연구)

  • Gong, Bokyoung;Jo, Young-Hoon;Ju, Soyeong;Min, Ji-Sook;Kwon, Mia
    • Analytical Science and Technology
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    • v.33 no.3
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    • pp.151-158
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    • 2020
  • Alcohol, which can easily be obtained in the same way as ordinary beverages, is harmful enough to cause death due to excessive drinking and chronic alcohol intake, so it is important to maintain a proper amount of drinking and healthy drinking habits. In addition, the incidence of behavioral disturbances and impaired judgments that can be caused by chronic alcohol drinking of more than adequate amounts of alcohol is also significant. Accordingly it is very useful for forensic science to check whether the person involved is drunken or is alcoholism state in various accidents. Currently, in Korea, alcohol consumption is determined by detecting the level of alcohol or alcohol metabolism 'ethyl glucuronide (EtG)' in blood or urine samples. However, analysis of alcohol or EtG in blood or urine can only provide information about the current state of alcohol consumption because of a narrow window of detection time. Therefore, it is important to analyze the EtG as a long-term direct alcohol metabolite bio-marker in human hair and to investigate relationship between alcohol consumption and EtG concentration for the evaluation of chronic ethanol consumption. In this study, we established an analytical method for the detection of EtG in Korean hair efficiently and validated selectivity, linearity, limits of detection (LOD), limits of quantification (LOQ), matrix effect, recovery, process efficiency, accuracy and precision using liquid chromatography tandem mass spectrometry (LC-MS/MS). In addition, the assay performance was evaluated in Korean social drinker's hair and the postmortem hair of a chronic alcoholism. The results of this study can be useful in monitoring the alcohol abuse of Korean in clinical cases and legal procedures related to custody and provide a useful tool to evaluate postmortem diagnosis of alcoholic ketoacidosis in forensics.

Functional analysis of seaR protein identified from Saccharopolyspora erythraea (희소방선균의 seaR 단백질 발현을 통한 기능 분석)

  • Ryu, Jae Ki;Kwon, Pil-Seung;Lee, Hyeong Seon
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.39-47
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    • 2015
  • Secondary metabolism in actinomycetes has been known to be controlled by a small molecule, ${\gamma}$-butyrolactone autoregulator, the binding of which to each corresponding receptor leads to the regulation of the transcriptional expression of the secondary metabolites. We expected that expression of an autoregulator receptor or a pleiotropic regulator in a non-host was to be gained insight of effective production of new metabolic materials. In order to study the function of the receptor protein (seaR), which is isolated from Saccharopolyspora erythraea, we introduced the seaR gene to Streptomyces coelicolor A3(2) as host strains. An effective transformation procedure for S. coelicolor A3(2) was established based on transconjugation by Escherichia coli ET12567/pUZ8002 with a ${\varphi}C31$-derived integration vector, pSET152, which contained int, oriT, attP and $ermEp^*$ (erythromycin promotor). Therefore, the pEV615 was introduced into S. coelicolor A3(2) by conjugation and integrated at the attB locus in the chromosome of the recipients by the ${\varphi}C31$ integrase (int) function. Exconjugant of S. coelicolor A3(2) containing the seaR gene was confirmed by PCR and transcriptional expression of the seaR gene in the transformant was analyzed by RT-PCR. In case of S. coelicolor A3(2), a phenotype microarray was used to analyze the phenotype of transformant compared with wild type by seaR expression. After that, in order to confirm the accuracy of the results obtained from the phenotype microarray, an antimicrobial susceptibility test was carried out. This test indicated that sensitivity of the transformant was higher than wild type in tetracycline case. These results indicated that some biosynthesis genes or resistance genes for tetracycline biosynthesis in transformant might be repressed by seaR expression. Therefore, subsequent experiments, analysis of transcriptional pattern of genes for tetracycline production or resistance, are needed to confirm whether biosynthesis genes or resistance genes for tetracycline are repressed or not.

Metabolizing analysis according to the sawdust media of the known anticancer trees by Pleurotus ostreatuss (느타리버섯의 항암수목자원 배지속 함유성분의 분해능 평가)

  • Shin, Yu-Su;Yang, Bo-Hyun;Kang, Bo-Yeon;Kim, Hyun-Soo;Lee, Ji-Hyun;Hong, Yoon-Pyo;Lee, Sang-Won;Lee, Chan-Jung;Kim, Seung-Yoo
    • Journal of Mushroom
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    • v.9 no.4
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    • pp.186-189
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    • 2011
  • The transitivity of Chemical constituents by Pleurotus ostreatus cultivated in different raw sawdusts, which are Juglans mandchurica, Cudrania tricuspidata and Lindera glauca, was investigated. The HPLC chromatography patterns on the chemical constituents of P. ostreatus showed the similar chromatography patterns in all different raw sawdusts and control sawdust. The unknown chemical constituents of P. ostreatus cultivated in the 10%, 20% mixed medium added 10 %, 20% different raw sawdusts, respectively, were increased. But the significance results in the mixed medium added 50% different raw sawdusts were not showed. The chromatography patterns of mycelia grown in media added the 80% MeOH extracts of three tree species showed the similar patterns in comparison with control mycelia. In the results, the secondary metabolites of functional media were not degrade and changed to other derivatives compounds by P. ostreatus.

Metabolizing analysis according to the sawdust media of Pleurotus ostreatus (산겨릅나무와 옻나무 톱밥배지에서 재배한 느타리버섯의 함유성분 분석)

  • Shin, Yu-Su;Yang, Bo-Hyun;Kang, Bo-Yeon;Kim, Hyun-Soo;Lee, Ji-Hyun;Hong, Yoon-Pyo;Lee, Sang-Won;Lee, Chan-Jung;Kim, Seung-Yoo
    • Journal of Mushroom
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    • v.9 no.4
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    • pp.194-197
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    • 2011
  • The transitivity of Chemical constituents by Pleurotus ostreatus cultivated in different raw sawdusts, which are Acer tegmentosum MAX, Rhus verniciflura, was investigated. The HPLC chromatography patterns on the chemical constituents of P. ostreatus showed the similar chromatography patterns in the different raw sawdusts and control sawdust. The unknown chemical constituents of P. ostreatus cultivated in the mixed medium added 10 %, 20% raw sawdusts, respectively, were increased. But the significance results in the mixed medium added 50% raw sawdusts were not showed. The chromatography patterns of mycelia grown in media added the 80% MeOH extracts of A. tegmentosum and R. verniciflura showed the similar patterns in comparison with control mycelia. In the results, the secondary metabolites of functional media were not degrade and changed to other derivatives compounds by P. ostreatus.

Effects of Dietary Quercetin on Growth Performance, Blood Biochemical Parameter, Immunoglobulin and Blood Antioxidant Activity in Broiler Chicks (사료 내 Quercetin의 첨가가 육계의 생산성, 혈액 생화학 특성, 혈액 내 면역글로불린과 혈액 내 항산화 인자에 미치는 영향)

  • Kim, Dong-Wook;Hong, Eui-Chul;Kim, Ji-Hyuk;Bang, Han-Tae;Choi, Ji-Young;Ji, Sang-Yoon;Lee, Wang-Shik;Kim, Sang-Ho
    • Korean Journal of Poultry Science
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    • v.42 no.1
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    • pp.33-40
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    • 2015
  • This study was conducted to investigate the effects of dietary quercetin on growth performance, blood biochemical parameters, immunoglobulin, and blood antioxidant activity in broiler chickens. Three hundred twenty one-day old Ross broilers were divided 8 treatments (C(-), basal diet; C(+), basal diet with antibiotics; vitamin E 20 IU; vitamin E 200 IU; quercetin 20 ppm; quercetin 200 ppm; methoxylated quercetin 20 ppm; methoxylated quercetin 200 ppm) with 4 replicates and 10 birds per replicate. Birds were reared for 35 days and their feed intake and weight gain were measured weekly. At 35d, eight birds of average weight from each replicate were selected for blood collection and analysis. Weight gain of birds in the groups fed quercetin was higher when compare to NC but there was no significant difference. In the serum, creatinine, BUN and AST in quercetin groups significantly decreased compared to those of control (NC and PC) (P<0.05). The contents of IgA and IgM were significantly lower in quercetin groups than those of NC (P<0.05). SOD like activity and MDA content tended to decrease in quercetin groups, however, there was no significant difference among treatments. In conclusion, supplemental quercetin to poultry diet could be positive aspect on performance and blood metabolites. Optimum adding levels was more than 20 ppm.

Determination of methamphetamine, 4-hydroxymethamphetamine, amphetamine and 4-hydroxyamphetamine in urine using dilute-and-shoot liquid chromatography-tandem mass spectrometry (시료 희석 주입 LC-MS/MS를 이용한 소변 중 메스암페타민, 4-하이드록시메스암페타민, 암페타민 및 4-하이드록시암페타민 동시 분석)

  • Heo, Bo-Reum;Kwon, NamHee;Kim, Jin Young
    • Analytical Science and Technology
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    • v.31 no.4
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    • pp.161-170
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    • 2018
  • The epidemic of disorders associated with synthetic stimulants, such as methamphetamine (MA) and amphetamine (AP), is a health, social, legal, and financial problem. Owing to the high potential of their abuse and addiction, reliable analytical methods are required to detect and identify MA, AP, and their metabolites in biological samples. Thus, a dilute-and-shoot liquid chromatography-tandem mass spectrophotometry (LC-MS/MS) was developed for simultaneous determination of MA, 4-hydroxymethamphetamine (4HMA), AP, and 4-hydroxyamphetamine (4HA) in urine. Urine sample ($100{\mu}L$) was mixed with $50{\mu}L$ of mobile phase consisting of 0.4 % formic acid and methanol and $50{\mu}L$ of working internal-standard solution. Aliquots of $8{\mu}L$ diluted urine was injected into the LC-MS/MS system. For all analytes, chromatographic separation was performed using a C18 reversed-phase column with gradient elution and a total run time of 5 min. The identification and quantification were performed by multiple reaction monitoring (MRM). Linear least-squares regression was conducted to generate a calibration curve, with $1/x^2$ as the weighting factor. The linear ranges were 2.0-200, 1.0-800, and 10-2500 ng/mL for 4HA and 4HMA, AP, and MA, respectively. The inter- and intraday precisions were within 6.6 %, whereas the inter- and intraday accuracies ranged from -14.9 to 11.3 %. The low limits of quantification were 2.0 ng/mL (4HA and 4HMA), 1.0 ng/mL (AP), and 10 ng/mL (MA). The proposed method exhibited satisfactory selectivity, dilution integrity, matrix effect, and stability, which are required for validation. Moreover, the purification efficiency of high-speed centrifugation was clearly higher than 6-15 % for QC samples (n=5), which was higher than that of the membrane-filtration method. The applicability of the proposed method was tested by forensic analysis of urine samples from drug abusers.

Alteration Analysis of Normal Human Brain Metabolites with Variation of SENSE and NEX in 3T Multi Voxel Spectroscopy (3T Multi Voxel Spectroscopy에서 SENSE와 NEX 변화에 따른 정상인 뇌 대사물질 변화 분석)

  • Seong, Yeol-Hun;Rhim, Jae-Dong;Lee, Jae-Hyun;Cho, Sung-Bong;Woo, Dong-Chul;Choe, Bo-Young
    • Progress in Medical Physics
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    • v.19 no.4
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    • pp.256-262
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    • 2008
  • To evaluate the metabolic changes in normal adult brains due to alterations SENSE and NEX (number of excitation) by multi voxel MR Spectroscopy at 3.0 Tesla. The study group was composed of normal volunteers (5 men and 8 women) with a mean ($\pm$ standard deviation) age of 41 (${\pm}11.65$). Their ages ranged from 28 to 61 years. MR Spectroscopy was performed with a 3.0T Achieva Release Version 2.0 (Philips Medical System-Netherlands). The 8 channel head coil was employed for MRS acquisition. The 13 volunteers underwent multi voxel spectroscopy (MVS) and single voxel spectroscopy (SVS) on the thalamus area with normally gray matter. Spectral parameters were as follows: 15 mm of thickness; 230 mm of FOV (field of view); 2000 msecs of repetition time (TR); 288 msecs of echo time (TE); $110{\times}110$ mm of VOI (view of interest); $15{\times}15{\times}15$ mm of voxel size. Multi voxel spectral parameters were made using specially in alteration of SENSE factor (1~3) and 1~2 of NEX. All MRS data were processed by the jMRUI 3.0 Version. There was no significant difference in NAA/Cr and Cho/Cr ratio between MVS and SVS likewise the previous results by Ross and coworkers in 1994. In addition, despite the alterations of SENSE factor and NEX in MVS, the metabolite ratios were not changed (F-value : 1.37, D.F : 3, P-value : 0.262). However, line-width of NAA peak in MVS was 3 times bigger than that in SVS. In the present study, we demonstrated that the alterations of SENSE factor and NEX were not critically affective to the result of metabolic ratios in the normal brain tissue.

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A case-control study on the effects of the genetic polymorphisms of N-acetyltransferase 2 and glutathione S-transferase mu and theta on the risk of bladder cancer (N-Acetyltransferase 2와 glutathione S-transferase mu 및 theta 다형성이 방광암 발생에 미치는 영향에 대한 환자-대조군 연구)

  • Kim, Heon;Kim, Wun-Jae;Lee, Hyung-Lae;Lee, Moo-Song;Kim, Cheol-Hwan;Kim, Ro-Sa;Nan, Hong-Mei
    • Journal of Preventive Medicine and Public Health
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    • v.31 no.2 s.61
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    • pp.275-284
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    • 1998
  • Activities of enzymes involved in the metabolism of various carcinogenic xenobiotics is one of the most important host factors for cancer occurrence. N-acetyltransferase (NAT) and glutathione S-transferases (GST) are enzymes which .educe the toxicity of activated carcinogenic metabolites. Slow N-acetylation and lack of GST mu (GSTMI) were reported as risk factors of bladder cancer. GST theta (GSTT1), which is another type of GST, was reported to be deleted at higher proportion among Koreans. Since cause of bladder cancer is not fully explained by single risk factor, many kinds of enzymes would be involved in the metabolism of carcinogens excreted in urine. This study was performed to investigate whether the polymorphisms of NAT2, GSTM1 and GSTT1 are risk factors of bladder cancer and to evaluate the effects of their interaction on bladder cancer development. Sixty-seven bladder cancer and 67 age- and sex-matched non-cancer patients hospitalized in Chungbuk National University Hospital from March to December 1996, are the subjects of this case-control study. Questionnaire interview was done and the genotypes of NAT2, GSTM1 and GSTT1 were identified using PCR methods with DNA extracted from venous blood. The effects of the polymorphism of NAT2 and GSTM1 and their interaction on bladder cancer were statistically tested after controlling the other risk factors. The frequencies of slow, intermediate, and rapid acetylators were 3.0%, 38.8%, and 58.2% to. the cases, and 7.6%, 40.9%, and 51.5% for the controls, respectively. The risk of bladder cancer was not associated with the increase of NAT2 activity($\chi^2_{trend}=1.18$, P-value>0.05). GSTM1 was deleted in 68.7% of the cases and 49.3% of the controls ($\chi^2=5.21$, P-value<0.05), and the odds ratio (95% CI) was 2.23 (1.12 - 4.56). GSTT1 deletion, the .ate of which were 26.9% for the bladder cancer patients and 43.3% for the controls, was a significant protective factor against bladder cancer. Smoking history turned out to be insignificant as a risk factor of bladder cancer (OR=1.85, 95% CI: 0.85 - 4.03), and occupation could not be tested because of the extremely small number of occupational history related to the increase of bladder cancer. In multiple logistic analysis controlling the effects of other risk factors, GSTM1 deletion was the only significant risk factor for bladder cancer (OR: 2.56, 95% CI: 1.22-5.36, P-value<0.05), but slow acetylation and GSTT1 deletion were not. These results suggest that GSTM1 deletion may be a significant risk factor of bladder cancer. Since there have been much debates on causal relationship between slow acetylation and GSTT1 deletion, and bladder cancer, further studies are needed.

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