• Archives of Pharmacal Research
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• 제29권10호
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• pp.911-920
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• 2006

Phenolic antioxidant butylated hydroxyanisole (BHA) is a commonly used food preservative with broad biological activities, including protection against chemical-induced carcinogenesis, acute toxicity of chemicals, modulation of macromolecule synthesis and immune response, induction of phase II detoxifying enzymes, as well as its undesirable potential tumor-promoting activities. Understanding the molecular basis underlying these diverse biological actions of BHA is thus of great importance. Here we studied the pharmacokinetics, activation of signaling kinases and induction of phase II/III drug metabolizing enzymes/transporter gene expression by BHA in the mice. The peak plasma concentration of BHA achieved in our current study after oral administration of 200 mg/kg BHA was around $10\;{\mu}M$. This in vivo concentration might offer some insights for the many in vitro cell culture studies on signal transduction and induction of phase II genes using similar concentrations. The oral bioavailability (F) of BHA was about 43% in the mice. In the mouse liver, BHA induced the expression of phase II genes including NQO-1, HO-1, ${\gamma}-GCS$, GST-pi and UGT 1A6, as well as some of the phase III transporter genes, such as MRP1 and Slco1b2. In addition, BHA activated distinct mitogen-activated protein kinases (MAPKs), c-Jun N-terminal kinase (JNK), extracellular signal-regulated protein kinase (ERK), as well as p38, suggesting that the MAPK pathways may play an important role in early signaling events leading to the regulation of gene expression including phase II drug metabolizing and some phase III drug transporter genes. This is the first study to demonstrate the in vivo pharmacokinetics of BHA, the in vivo activation of MAPK signaling proteins, as well as the in vivo induction of Phase II/III drug metabolizing enzymes/transporters in the mouse livers.

• Journal of Microbiology and Biotechnology
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• 제14권3호
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• pp.584-592
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• 2004

A thermostable $\beta$-glycosidase gene, tfi $\beta$-gly, was cloned from the genomic library of Thermus filiformis Wai33 A1. ifi $\beta$-gly consists of 1,296 bp nucleotide sequence and encodes a polypeptide of 431 amino acids. It shares a strong amino acid sequence similarity with the $\beta$-glycosidases from other Thermus spp. belonging to the glycosyl hydrolase family 1. In the present study, the enzyme was overexpressed in Escherichia coli BL21 (DE3) using the pET21b(+) vector system. The recombinant enzyme was purified to homogeneity by heat treatment and a $Ni^{2+}$-affinity chromatography. Polyacrylamide gel electrophoresis (PAGE) showed that the recombinant Tfi $\beta$-glycosidase was a monomeric form with molecular mass of 49 kDa. The temperature and pH range for optimal activity of the purified enzyme were 80- $90^{\circ}C$ and 5.0-6.0, respectively. Ninety-three percent of the enzyme activity was remained at $70^{\circ}C$ after 12 h, and its half-life at $80^{\circ}C$ was 6 h, indicating that Tfi $\beta$-glycosidase is highly thermostable. Based on its K_m$, or $K_{cat}K_m$, ratio, Tfi $\beta$-glycosidase appeared to have higher affinity for $\beta$-D-glucoside than for $\beta$-D-galactoside, however, $K_{cat} for \beta$-D-galactoside was much higher than that for $\beta$-D-glucoside. The activity for lactose hydrolysis was proportionally increased at $70^{\circ}C$ and pH 7.0 without substrate inhibition until reaching 250 mM lactose concentration. The specific activity of Tfi TEX>$\beta$-glycosidase on 138 mM lactose at $70{^\circ}C$ and pH 7.0 was 134.9 U/mg. Consequently, this newly cloned enzyme appears to have a valuable advantage of conducting biotechnological processes at elevated temperature during milk pasteurization in the production of low-lactose milk.

• 한국미생물·생명공학회지
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• 제30권1호
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• pp.51-56
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• 2002

국내 토양에서 분리한 식물성 병원균인 Alternaria brassicicola SW-3리 항암활성 물질 생산능을 조사하고, 활성물질을 분리 정제하여 구조를 확인하였다. A. brassicicola SW-3는potato dextrose broth를 이용하여 15$^{\circ}C$에서 2주간 진탕 배양한 다음, MTT assay를 실시하여 항암활성을 확인하였으며, 배양여액 중의 항암물질은 ethyl acetate로 추출하고, silica gel column chromatography로 정제하여 무색의 oily product를 얻었다(수율 22mg/m1). 분리된 물질은 물이나 hexane에는 녹지 않고, chloroform, ethyl acetate, ethanol 등에는 잘 녹는 특징을 보였으며, , $IR^{1}$H-NMR, $^{13}$C-NMR 등을 통해 구조를 분석한 결과, 최근에 일본에서 분리되어 항암효과가 있는 것으로 알려진 depudecin과 동일한 물질로 추정되었다. 본 실험에서 분리된 depudecin은 인체간암세포와 mouse 피부암세포에 대한 세포독성을 나타내었으며, 각각의$ IC_50$ 는 $57\mu$g/ml, $69\mu$g/ml로 나타났다. Alternaria brassicicola SW-3에 의해 생산된 물질이 기지의 물질이지만, depudecin은 아직 작용 기작이나 적용범위 등이 밝혀지지 않은 초기 연구 단계에 있는 물질로서, 새로운 항암제로서의 가능성이 매우 높아 이의 유도체를 합성하거나, 다른 항암제와의 혼용에 의해 부작용이 적은 강력한 항암제를 개발하기 위한 선도물질로 활용될 수 있을 것이다.

• 생약학회지
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• 제46권2호
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• pp.167-172
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• 2015

Aleurone layer of Black rice (Oryza sativa L.) is enriched with anthocyanin that could increase bone density. This study was conducted to investigate the osteoporosis-preventing effects of the aleurone layer extract (BRE) on bone loss of ovariectomized (OVX) rats. OVX (or sham-operated) rats were assigned to three groups (n=8 per group): sham operated group (Sham); OVX control group (OVX); OVX-BRE group, OVX rats treated with BRE at 90 mg/kg B.W. The deionized water alone or deionized water with BRE was orally administrated to Sham, OVX or OVX-BRE groups, respectively for 12 weeks. High fat diet with 45 kcal% fat and water were fed to all rats ad libitum. Body weight was significantly decreased in the OVXBRE group compared to the OVX group (p<0.05). The bone mineral density and bone length of tibia were significantly higher in the OVX-BRE group compared to the OVX group and breaking force was significantly higher for the both tibia and femur bones. Serum estradiol concentration and calcium concentration of femur were higher in the OVX-BRE group than those of OVX group. However, serum alkaline phosphatase activity and parathyroid hormone concentration were decreased in the OVXBRE group compared to the OVX group. The results suggest that aleurone layer of Black rice is a potentially useful ingredient to protect against estrogen deficiency or menopause related osteoporosis.

• 한국식품저장유통학회지
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• 제19권6호
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• pp.849-857
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• 2012

전처리 방법에 따른 적외선건조 복숭아의 이화학적 및 기능적 특성을 조사한 결과 수분함량은 0.3% NaCl 용액 침지구에서 낮게 나타났다. 가용성 고형분 함량은 vitamin C와 soluble Ca 처리 농도가 증가함에 따라 유의적으로 증가하는 경향을 보였다. 식미와 밀접한 관계가 있는 당산비는 0.1% vitamin C처리구와 2.0% soluble Ca처리구에서 높은 값을 나타내었다. 색도는 각각의 전처리구에 있어서 $L^*$ 값 및 $a^*$ 값은 높고 $b^*$ 값은 낮은 경향을 나타내어 갈변이 억제되는 결과를 보였다. 절단력은 전처리구가 무처리구에 비하여 높게 나타났으며, soluble Ca 처리에 따른 무기질 함량은 K > Mg > Ca > Na > Fe > Zn > Cu의 순을 나타내었으며 soluble Ca의 처리 농도가 증가됨에 따라 무기질 함량과 Ca의 함량 또한 증가하는 것으로 나타났다. 관능특성에서는 각각의 전처리 구가 무처리구에 비하여 전반적으로 기호도가 높아지는 결과를 나타내었다. 전처리방법에 따른 적외선건조 복숭아의 총 폴리페놀 및 플라보노이드 함량은 전반적으로 무처리구에 비해 0.1% vitamin C처리구, 1.0% soluble Ca 처리구에서 높게 나타났으며 ABTS 라디칼 소거 활성에서도 이와 유사한 경향을 나타내었다. 이상의 결과에서 이화학적, 기능적 품질특성을 고려해 보았을 때 0.1% vitamin C 전처리와 적외선건조방법을 효과적으로 이용하는 것이 고품질 복숭아 건조제품을 제조할 수 있는 방안이 될 것으로 사료된다.

• 한국식품저장유통학회지
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• 제24권3호
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• pp.421-430
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• 2017

본 연구에서는 표고버섯 분말을 4%, 8%, 12%로 첨가하여 쌀 쿠키를 제조하고 품질 특성, 항산화 활성 및 기호도 조사를 실시하여 기능성 쿠키를 개발하고자 하였다. 쿠키반죽의 밀도는 표고버섯 분말 첨가량이 증가함에 따라 증가하였고 pH는 감소하였다. 쿠키의 수분 함량은 대조군이 8.32%로 가장 높았으며 첨가군은 4.73-5.56%로 대조군보다 낮았다. 표고버섯 분말 첨가량이 증가함에 따라 퍼짐성, 손실률, 팽창률 모두 감소하였다. 쿠키의 색도는 표고버섯 분말 첨가량이 증가함에 따라 L 값은 감소하였고 a 값은 증가하였으며 b 값은 대조군이 가장 높았다. 경도는 대조군이 가장 낮았고 12% 첨가군이 가장 높았다. 쿠키의 미세구조는 표고버섯 분말 첨가량이 증가할수록 표면과 내부 조직이 치밀해졌다. 총 폴리페놀 함량은 9.78-18.18 mg GAE/100 g의 범위로 표고버섯 분말 첨가량이 증가함에 따라 증가하였고, DPPH radical 소거능과 환원력도 증가하였다. 기호도 조사 결과, 색은 대조군이 높은 기호도를 나타내었으며 냄새, 맛, 전체적인 기호도는 대조군을 포함한 모든 시료들 간에 유의적인 차이가 없었다. 이러한 결과로 보아 쿠키에 표고버섯 분말을 첨가하는 것은 쿠키의 기호도를 증가시키고 동시에 항산화 활성을 증가시킴으로써 쿠키의 가치를 높여 줄 것으로 기대되며, 쿠키 제조 시 표고버섯 분말을 12% 첨가하는 것이 바람직할 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제29권4호
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• pp.651-657
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• 2019

Although smallpox was eradicated in 1980, it is still considered a potential agent of biowarfare and bioterrorism. Smallpox has the potential for high mortality rates along with a major public health impact, eventually causing public panic and social disruption. Passive administration of neutralizing monoclonal antibodies (mAbs) is an effective intervention for various adverse reactions caused by vaccination and the unpredictable nature of emerging and bioterrorist-related infections. Currently, vaccinia immune globulin (VIG) is manufactured from vaccinia vaccine-boosted plasma; however, this production method is not ideal because of its limited availability, low specific activity, and risk of contamination with blood-borne infectious agents. To overcome the limitations of VIG production from human plasma, we isolated two human single-chain variable fragments (scFvs), (SC34 and SC212), bound to vaccinia virus (VACV), from a scFv phage library constructed from the B cells of VACV vaccine-boosted volunteers. The scFvs were converted to human IgG1 (VC34 and VC212). These two anti-VACV mAbs were produced in Chinese Hamster Ovary (CHO) DG44 cells. The binding affinities of VC34 and VC212 were estimated by competition ELISA to $IC_{50}$ values of $2{\mu}g/ml$ (13.33 nM) and $22{\mu}g/ml$ (146.67 nM), respectively. Only the VC212 mAb was proven to neutralize the VACV, as evidenced by the plaque reduction neutralization test (PRNT) result with a $PRNT_{50}$ of ~0.16 mg/ml (${\sim}1.07{\mu}M$). This VC212 could serve as a valuable starting material for further development of VACV-neutralizing human immunoglobulin for a prophylactic measure against post-vaccination complications and for post-exposure treatment against smallpox.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• 제25권3호
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• pp.163-179
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• 2022

Although functional gastrointestinal disorders (FGIDs) are very common in pediatric patients, there is a scarcity of published epidemiologic data, characteristics, and management patterns from Saudi Arabia, which is the 2nd largest Arabic country in terms of area and the 6th largest Arabic country in terms of population, with 10% of its population aged <5 years. Functional constipation (FC) is an FGID that has shown a rising prevalence among Saudi infants and children in the last few years, which urges us to update our clinical practices. Nine pediatric consultants attended two advisory board meetings to discuss and address current challenges, provide solutions, and reach a Saudi national consensus for the management of pediatric constipation. The pediatric consultants agreed that pediatricians should pay attention to any alarming signs (red flags) found during history taking or physical examinations. They also agreed that the Rome IV criteria are the gold standard for the diagnosis of pediatric FC. Different therapeutic options are available for pediatric patients with FC. Dietary treatment is recommended for infants with constipation for up to six months of age. When non-pharmacological interventions fail to improve FC symptoms, pharmacological treatment with laxatives is indicated. First, the treatment is aimed at disimpaction to remove fecal masses. This is achieved by administering a high dose of oral polyethylene glycol (PEG) or lactulose for a few days. Subsequently, maintenance therapy with PEG should be initiated to prevent the re-accumulation of feces. In addition to PEG, several other options may be used, such as Mg-rich formulas or stimulant laxatives. However, rectal enemas and suppositories are usually reserved for cases that require acute pain relief. In contrast, infant formulas that contain prebiotics or probiotics have not been shown to be effective in infant constipation, while the use of partially hydrolyzed formula is inconclusive. These clinical practice recommendations are intended to be adopted by pediatricians and primary care physicians across Saudi Arabia.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.51-51
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• 2001

Cocaine and amphetamine-regulated transcript (CART) is a satiety factor that is regulated by leptin. It was reported that the mice intracerebroventricularly injected with CART showed behavioral changes resembled with the typical behavioral alterations found in the mice carrying disorders in the brain serotonergic (5-HT) system. Hence, this study was conducted to find out the relationships between CART and 5-HT. We first examined the mRNA levels of CART after the injections of para-chlorophenylalanine (pCPA, 300 mg/kg i.p., single injection or daily for three consecutive days) in the rat brains by in situ hybridization using the mouse CART cDNA probe cloned in our laboratory. Systemic administrations of pCPA, a potent inhibitor of tryptophan hydroxylase, the rate limiting enzyme of 5-HT biosynthesis, acutely depletes the brain 5-HT transporter (5-HTT) in the dorsal raphe nucleus (DRN), which reuptakes terminal 5-HT. Results indicated that the mRNA level of CART significantly decreased in the arcuate nucleus, paraventricular nucleus, and lateral hypothalamic nucleus by three days of daily injection with pCPA with no noticeable change detected 24 hrs after the single injection. The message levels of 5-HTT in DRN decreased in both single and three days of injections. Secondly, to investigate whether CART affect to 5-HT, mouse genomic CART gene, which is consist of 3 exons and 2 introns and mouse neurofilament light (NF-L) chain promoter were cloned. Then, we constructed neuron specific expression vector, which was transfected into HeLa cell using lipid-mediated transfection system. Expression of GFP and CART linked to NF-L-chain promoter in the transfected HeLa cell were detected by using fluorescent microscope and RT-PCR. These results confirmed normal expression of DNA constructs in vitro. Then, to increase brain specific expression of CART in vivo transgenic mice carrying CART gene controlled the deleted NF-L-chain promoter were generated by the DNA microinjection into pronuclei of fertilized embryos. Transgenic mice were detected by Southern blot. Further study is necessary to examine CART expression and 5-HTT in these transgenic mice. Therefore, these results suggest that there maybe a positive molecular correlation between CART and 5-HT in responding to the stimuli.

• Annals of Occupational and Environmental Medicine
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• 제35권
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• pp.38.1-38.10
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• 2023

Background: Hearing loss (HL) is linked to an elevated risk of cardiovascular diseases (CVDs). The pathogeneses of HL and CVD commonly involve inflammatory responses. Previous studies investigated elevated levels of inflammatory biomarkers in subjects with HL, however, their findings did not demonstrate statistical significance. In our cross-sectional and longitudinal study, we investigated the correlation between HL and increased high-sensitivity C-reactive protein (hsCRP) levels to determine how HL is associated with CVDs. Methods: We conducted a cross-sectional study with workers aged over 18 years who underwent health check-ups at our institution between 2012 and 2018 (n = 566,507), followed by conducting a longitudinal study of workers aged > 18 who underwent health checkups at least twice at our institution between 2012 and 2018 (n = 173,794). The definition of HL was as an average threshold of ≥ 20 dB in pure-tone air conduction at 0.5, 1.0, and 2.0 kHz in both ears. The incidence of increased hsCRP levels throughout the follow-up period was defined as a level exceeding 3 mg/L. Logistic regression and generalized estimating equations were performed to estimate the risk of increased hsCRP levels according to the occurrence of HL in groups stratified by age. Results: In the cross-sectional study, the multivariate-adjusted odds ratio (OR) was 1.17 (95% confidence interval [CI]: 1.02-1.34); the OR was 0.99 (95% CI: 0.80-1.22) in those under 40 and 1.28 (1.08-1.53) in those over 40. In the longitudinal study, the multivariable-adjusted OR was 1.05 (95% CI: 0.92-1.19); the OR was 1.10 (95% CI: 0.90-1.35) in those under 40 and 1.20 (1.01-1.43) in those over 40. Conclusions: This cross-sectional and longitudinal study identified an association between HL and increased hsCRP levels in workers aged over 40 years.