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Studies on Biological Characteristics of Lactobacillus II. Conjugal Transfer-frequency of R Plasmids from Lactobacillus to Escherichia coli (유산균(乳酸菌)의 생물학적특성(生物學的特性)에 관한 연구(硏究) II. 약제내성(藥劑耐性) 유산균(乳酸菌)의 R Plasmids 전달빈도(傳達頻度))

  • Kim, Jong Myeon;Song, Hee Jong
    • Korean Journal of Veterinary Research
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    • v.20 no.2
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    • pp.113-118
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    • 1980
  • Total of 11 strains of Ldctobacillus isolated from lactobacillus-fermented milk and-beverage in March 1980 were examined for susceptibility to 8 drugs, and transferability and transfer frequency of R plasmids by conjugation. Of 11 isolates each 2 strains were classified as L. cellobiosus and L. helveticus, each 1 strain as L. plantarum, L. lactis, L. acidophilus, L. delbrueckii, L. casei subsp. casei, L. casei subsp, tolerans and L. salivarius subsp, salivarius by Bergey's manual. Resistance was the most active to na lidixic acid(NA), followed in decreasing order by chloramphenicol(CP), ampicillin(AP), kanamycin(KM) and streptomycin(SM). All of isolates were resistant to NA, each 10 strains to CP and AP, 7strains to KM and 6 strains to SM, indicating all of the isolates were resistant to two or more drugs in combination. No strain was resistant to erythromycin(EM), penicillin(PC) and tetracycline(TC). The most frequently encountered resistant patterns were CP NA AP SM KM, followed by CP NA AP KM, NA AP, CP NA, CP NA AP and CP NA AP SM in order. Transfer experiment of drug resistance showed that of 11 resistant strains, 9 strains transferred parts of their resistance to AP or AP CP or SM AP, indicating 9 strains carried R plasmids determining R(AP), R(AP CP) and R(SM AP). The conjugal frequency of R(AP) from Lactobacillus to E. coli ranged from 2.5{\times}10^{-1} to $5.6{\times}10^{-4}%$, that of R(CP) ranged from 5.0{\times}10^{-1} to 5.0{\times}10^{-3}% and that of R(SM) ranged from 6.0{\times}10^{-5} to 1.4{\times}10^{-5}%, at $37^{\circ}C$ for 18 hours of incubation.

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Genome Wide Expression Profile of Agrimonia pilosa in LPS-stimulated BV-2 Microglial Cells

  • Sohn, Sung-Hwa;Ko, Eun-Jung;Kim, Sung-Hoon;Kim, Yang-Seok;Shin, Min-Kyu;Hong, Moo-Chang;Bae, Hyun-Su
    • Molecular & Cellular Toxicology
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    • v.5 no.1
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    • pp.1-6
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    • 2009
  • Microglial cells constitute the first line of defense against infection and injury in the brain. This study was conducted to evaluate the protective mechanisms of Agrimonia pilosa (AP) on LPS-induced activation of BV-2 microglial cells. The effects of AP on gene expression profiles in activated BV-2 microglial cells were evaluated using microarray analysis. BV-2 microglial cells were cultured in a 100 mm dish ($1{\times}10^7/mL$) for 24 hr and then pretreated with 1 g/mL AP or left untreated for 30 min. Next, 1 g/mL LPS was added to the samples and the cells were reincubated at $37^{\circ}C$ for 30 min, 3 hr and 6 hr. The gene expression profiles of the BV-2 microglial cells varied depending on the AP. The microarray analysis revealed that MAPK signaling pathway-related genes were down-regulated and IL10 gene was up-regulated in AP-treated BV-2 microglial cells. AP can affect the inflammatory response and MAPK pathway in BV-2 microglial cells.

Comparison of Main Compounds and Physiological Activities of Anthriscus sylvestris (L.) Hoffm. Roots and Aerial Parts Extracts (전호 지하부와 지상부 추출물의 주요성분 및 생리활성 비교)

  • Kim, Sol;Kim, Ha-Rim;Kim, Sang-Jun;Kim, Seon-Young
    • Korean Journal of Pharmacognosy
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    • v.52 no.2
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    • pp.77-83
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    • 2021
  • The number of people suffering from diabetes have been increased around the world. In this study, we investigated the antidiabetic and antioxidant effects of Anthriscus sylvestris(L.) Hoffm and its main compounds. It was divided into root(R) and aerial part(AP) for comparative analysis. Total polyphenol, total flavonoid content was higher in AP extract, but nodakenin content was higher in R(1169.13 ± 6.00 mg/g) extract. Antioxidant activity was also higher in AP extract. To compare antidiabetic efficacy, we analyzed the effects of R and AP extracts on ɑ-glucosidase inhibition(AGI), dipeptidyl peptidase-4(DPP-4) and protein tyrosine phosphatase(PTP)1B activity. R and AP extracts showed similar effects on AGI and DPP-4 activity in a concentration dependent manner, and there was no effect on PTP1B activation. Glucose uptake(139.51 ± 3.19%) in 3T3-L1 cells was more effective in the AP extract-treated group than the positive control, rosiglitazone, group. Both R and AP extracts were effective in protecting against pancreatic beta cell damage caused by hyperglycemia. These results suggest that Anthriscus sylvestris(L.) Hoffm. could be used as a candidate for diabetes treatment.

Drug Resistance and R Plasmids of Lactobacilli Isolated from Fermented Milk (유산균음료(乳酸菌飮料)로부터 분리(分離)한 유산간균(乳酸桿菌)의 R-Plasmids의 중개(仲介)에 의(依)한 대장균(大腸菌)에로의 항생제내성(抗生劑耐性) 전달(傳達))

  • Ha, Tai-You;Lee, Jeong-Ho
    • The Journal of the Korean Society for Microbiology
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    • v.15 no.1
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    • pp.55-62
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    • 1980
  • Eleven strains of lactobacilli were isolated from fermented milk of 9 companies. They were classified 3 strains as L. bulgaricus, 2 strains as L. plantarum, 2 strains as L. cellobiosus, 1 strain as L. lactis, 1 strain as L. acidophilus, 1 strain as L. casei subsp. casei and 1 strain as L. casei subsp. tolerans. And these strains were examined for drug resistance, transferability and transfer frequency of R plasmid. Most of isolates were sensitive to tetracycline(TC), penicillin(PC), and erythromycin(EM), but some strains were resistant to streptomycin(SM), chloramphenicol(CP), ampicillin(AP), kanamycin(KM), and nalidixic acid(NA). All of isolates were resistant to two or more drugs and 6 different drug resistant patterns were found. The most frequently encountered patterns were NA AP CP SM KM(5 strains) followed by NA AP CP KM(2 strains), NA AP CP SM(1 strain), NA AP CP(1 strain), NA CP(1 strain) and NA AP(1 strain). Tranfer experiment of drug resistance showed that of all 11 resistant strains, 9 strains transferred R plasmid determining AP(6 strains) or AP SM(3 strains) to a recipient, E. coli ML 1410 strain with $2.8{\times}10^{-5}-1.5{\times}10^{-1}%$ of transfer frequency. These results indicate that lactobacilli conjugally transfer their resistance to E. coli.

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Determination of methamphetamine and amphetamine enantiomers in human urine by chiral stationary phase liquid chromatography-tandem mass spectrometry

  • Sim, Yeong Eun;Ko, Beom Jun;Kim, Jin Young
    • Analytical Science and Technology
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    • v.32 no.5
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    • pp.163-172
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    • 2019
  • Methamphetamine (MA) is currently the most abused illicit drug in Korea and its major metabolite is amphetamine (AP). As MA exist as two enantiomers with the different pharmacological properties, it is necessary to determine their respective amounts in a sample. Thus a chiral stationary phase liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed for identification and quantification of d-MA, l-MA, d-AP, and l-AP in human urine. Urine sample ($200{\mu}L$) was diluted with pure water and purified using solid-phase extraction (SPE) cartridge. A $5-{\mu}L$ aliquot of SPE treated sample solution was injected into LC-MS/MS system. Chiral separation was carried out on the Astec Chirobiotic V2 column with an isocratic elution for each enantiomer. Identification and quantification of enantiomeric MA and AP was performed using multiple reaction monitoring (MRM) detection mode. Linear regression with a $1/x^2$ as the weighting factor was applied to generate a calibration curve. The linear ranges were 25-1000 ng/mL for all compounds. The intra- and inter-day precisions were within 3.6 %, while the intra- and inter-day accuracies ranged from -5.4 % to 11.8 %. The limits of detection were 2.5 ng/mL (d-MA), 3.5 ng/mL (l-MA), 7.5 ng/mL (d-AP), and 7.5 ng/mL (l-AP). Method validation parameters such as selectivity, matrix effect, and stability were evaluated and met acceptance criteria. The applicability of the method was tested by the analysis of genuine forensic urine samples from drug abusers. d-MA is the most common compound found in urine and mainly used by abusers.

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.

Expression of ATE2 Transcription Factor and the Interaction with AP-1 Factors : BATF, c-Fos, c-Jun (ATF2 전사인자의 발현과 AP-1 전사인자인 BATF, c-Fos, c-Jun과의 이량체 형성)

  • Jang Hye-Young;Kim Jae-Ho
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.928-934
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    • 2005
  • ATF2 is a cellular transcription factor which belongs to the CREB/ATF class and it is leucine zipper protein which generally binds to DNA as dimers. This paper presents the procedure for subcloning the ATF2 gene and the results of experiment used the expressed ATF2. The pET expression vector was used since it produced 6xHis fusion protein for easy purification using affinity column. The Nickel chelating chromatography was used for Purifying the expressed ATE2 from E- codi BL2l. Subsequen시y In vitro binding pull-down assay showed the binding specificity of ATF2 with AP-1 family factors such as BATF, c-Fos, c-Jun and ATF2 itselgf. ATF2 forms homodimer as well as strong heterodimer with BATF. It also forms stable dimer with c-Jun but barely binds with c-Fos.

Vasodilation Effect of the Water Extract of Gingko biloba, Acanthopanax senticosus and Augmented Four-Substance Decoctions in Rat Thoracic Aorta (백과엽(白果葉) 및 자오가(刺五加) 가미사물탕(加味四物湯)이 백서(白鼠)의 흉부대동맥(胸部大動脈) 혈관이완(血管弛緩)에 미치는 영향)

  • Kim Hyung-Hwan;Park Soo-Yeon;Kang Soon-Ah;Kim Hong-Yeoul;Ahn Duk-Kyun;Park Seong-Kyu
    • Herbal Formula Science
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    • v.11 no.2
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    • pp.125-134
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    • 2003
  • We have examined the relaxational response to the water extract of Angelica gigas $N_{AKAI}$ (AG), Gingko biloba $L_{INNE}$ (GB), Acanthopanax senticosus $H_{ARMS}.$ (AP) and Augumented-Four-Substance Decoction (AG-FSD, GB-FSD, AP-FSD) in isolated thoracic aorta from sprague dawley(SD) rat. Rat thoracic aorta was investigated in vessel segments suspended for isometric tension recording by polygraph. Responses to AG, GB, AP and AG-FSD, GB-FSD, AP-FSD were investigated in vessels precontracted with 5-hydroxytryptamine(5-HT) were compared in vasodilation effect. We found that the thoracic aorta segments responded to AG, GB, AP and AG-FSD, GB-FSD, AP-FSD with a dose-dependent vasodilation. The 5-hydroxytryptamine induced contraction at $10^{-4}M$ were inhibited by 26.3%, 75.8%, 87.5% and 6.9%, 22.6%, 30.8% after addition of the 0.1 g/mL water extract of AG, GB, AP and AG-FSD, GB-FSD, AP-FSD. In conclusion, AG, GB, AP and AG-FSD, GB-FSD, AP-FSD induced relaxation in the isolated rat thoracic aorta were composed of dose-dependent relaxation. AP-FSD has very potent vasodilation.

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Formulation Design of Sustained-Release Matrix Tablets Containing 4-Aminopyridine (유드라짓과 알긴산 나트륨 매트릭스를 이용한 4-Aminopyridine의 서방성 제제설계)

  • Kim, Jeong-Soo;Kim, Dong-Woo;Lee, Gye-Won;Jee, Ung-Kil
    • Journal of Pharmaceutical Investigation
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    • v.35 no.6
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    • pp.453-460
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    • 2005
  • 4-Aminopyridine (AP) is a potassium channel blocker used in the treatment of neurological disorders such as multiple sclerosis and Alzheimer disease. AP‘s window of therapeutic effect appears to correlate with its plasma halflife (3.5 hours). It demonstrates pH-dependent solubility because of a weakly basic drug. In addition, the resulting release from conventional matrix tablets decreases with increasing pH-milieu of the gastrointestinal tract. The aim of this study is to design sustained release matrix tablet containing AP, overcoming this problem. $Eudragit^{\circledR}$ L 100 (EuL) and sodium alginate were used in an effort to achieve pH independent drug release. The effect of sodium alginate and EuL on drug release from matrix tablet was investigated. The drug release behavior from the different tablets was analyzed by $t_{20%},\;t_{40%},\;t_{60%}$, The exponential diffusion coefficient n, kinetic constant K were calculated according to the Korsmeyer-Peppas equation. The drug release from matrix tablets prepared with sodium alginate was decreased with increasing the content of sodium alginate in pH 7.4 while there is no significant difference in pH 1.2. The exponent n values were determined to be approximately 0.5 and 0.8 respectively, in both pH 1.2 and 7.4. These values indicate diffusion-based anomalous mechanism and erosion-based anomalous mechanism, respectively. The drug release from sodium alginate matrix tablets prepared with solid dispersion of EuL containing drug showed a slow drug release in an acidic medium and a more fast drug release in phosphate medium, compared with sodium alginate matrix tablets prepared with physical mixture. These results may be attributed to the gel forming ability of sodium alginate and pH dependent solubility of EuL. Therefore, sustained-release AP matrix tablets using sodium alginate and EuL were successfully prepared.

Comparative study on growth of leafy vegetables grown in a hybrid BFT-aquaponics using Japanese eel, Anguilla japonica and hydroponics

  • Lee, Dong-Hoon;Kim, Jeong-Dae
    • Fisheries and Aquatic Sciences
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    • v.24 no.7
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    • pp.260-275
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    • 2021
  • Aquaponics is a cultivation system that combines aquaculture and agricultural hydroponics. This study investigated the productivity of leafy vegetables cultivated in the hybrid biofloc technology-aquaponics (HBFT-AP) using Japanese eel fed two kinds of diets (Mash, commercial powdered feed and extruded pellet [EP]) and hydroponics (HP).The mash was fed to the fish in a type of a dough mixed with water for 6 weeks (Exp1) and switched to the EP containing 2.7% monobasic potassium phosphate (MKP) for subsequent 6 weeks (Exp2). Leafy vegetables of 8 cultivar were employed in the experiment and water quality [dissolved oxygen (DO, mg/L), pH, water temperature (℃), electrical conductivity (EC; ㎲/cm), turbidity (NTU), TAN (NH3 + NH4+) (mg /L), NO2-N (mg/L), NO3-N (mg/L) and PO4-P (mg/L)] was measured 6 times a week. Leafy vegetable productivity (HBFT-AP vs HP) was compared in respective experiment, which was similar or somewhat higher in HBFT-AP. During the 12-week feeding trial, concentrations of nitrite (NO2-N) and phosphorus (PO4-P) were kept low in variability while total ammonia nitrogen (TAN) and NO3-N levels increased with time in HBFT-AP. At the end of two feeding trials, values of weight gain (WG, %), feed efficiency (FE, %), specific growth rate (SGR, %) and protein efficiency ratio (PER) were higher in Exp2 than in Exp1. As well, higher values in hematocrit (PCV, %), plasma K (mEq/L) and inorganic phosphorus (mg/dL) were found (p < 0.05) in Exp2 where fish were fed the EP (EP: 38.60%, 2.80 mEq/L and 7.04 g/dL; Mash: 33.20%, 1.95 mEq/L and 5.50 g/dL). Leafy vegetables in HBFT-AP using Japanese eel fed the EP with MKP 2.7% had a productivity similar (4 kinds of cultivar) or somewhat higher (4 kinds of cultivar) compared to those in HP. Also, Japanese eel fed the EP showed higher values of Pi (m/dL) and K (mEq/L) in plasma compared to those fed commercial powder diet.