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Gain of a New Exon by a Lineage-Specific Alu Element-Integration Event in the BCS1L Gene during Primate Evolution

  • Park, Sang-Je;Kim, Young-Hyun;Lee, Sang-Rae;Choe, Se-Hee;Kim, Myung-Jin;Kim, Sun-Uk;Kim, Ji-Su;Sim, Bo-Woong;Song, Bong-Seok;Jeong, Kang-Jin;Jin, Yeung-Bae;Lee, Youngjeon;Park, Young-Ho;Park, Young Il;Huh, Jae-Won;Chang, Kyu-Tae
    • Molecules and Cells
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    • v.38 no.11
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    • pp.950-958
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    • 2015
  • BCS1L gene encodes mitochondrial protein and is a member of conserved AAA protein family. This gene is involved in the incorporation of Rieske FeS and Qcr10p into complex III of respiratory chain. In our previous study, AluYRa2-derived alternative transcript in rhesus monkey genome was identified. However, this transcript has not been reported in human genome. In present study, we conducted evolutionary analysis of AluYRa2-exonized transcript with various primate genomic DNAs and cDNAs from humans, rhesus monkeys, and crabeating monkeys. Remarkably, our results show that AluYRa2 element has only been integrated into genomes of Macaca species. This Macaca lineage-specific integration of AluYRa2 element led to exonization event in the first intron region of BCS1L gene by producing a conserved 3' splice site. Intriguingly, in rhesus and crabeating monkeys, more diverse transcript variants by alternative splicing (AS) events, including exon skipping and different 5' splice sites from humans, were identified. Alignment of amino acid sequences revealed that AluYRa2-exonized transcript has short N-terminal peptides. Therefore, AS events play a major role in the generation of various transcripts and proteins during primate evolution. In particular, lineage-specific integration of Alu elements and species-specific Alu-derived exonization events could be important sources of gene diversification in primates.

Differential Diagnosis of Chemical-induced Hepatobiliary Toxicities Using a New Hepatobiliary Imaging Agent in Mice

  • Ryu, Chong-Kun;Pie, Jae-Eun;Choe, Jae-Gol;Cheon, Joon;Sohn, Jeong-Won;Jurgen Seidel;David S. Paik;Michael V. Green;Chang H. Paik;Kim, Meyoung-Kon
    • Environmental Mutagens and Carcinogens
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    • v.21 no.1
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    • pp.1-8
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    • 2001
  • We have synthesized $^{99m}$Tc-mercaptoacetyltriglycine (MAG3)-biocytin as a new imaging agent for hepatobiliary scintigraphy. The aim of this study was to evaluate the usefulness of $^{99m}$Tc-MAG3-biocytin scintigraphy in differentiating carbon tetrachloride ( $CCl_4$)-induced hepatotoxicity from $\alpha$-naphthylisothiocyanate (ANIT)-induced cholestasis in mice, which reflecting the differential diagnosis of neonatal jaundice caused by neonatal hepatitis from congenital biliary atresia in humans. Methods: Balb/c mice (female, 20 g, n=4-6) were pretreated with $CCl_4$(0.5 or $1.0m\ell$/kg) and ANIT ($150 or 300 m\ell$/kg) 18 h before scintigraphy. Biochemical and histopathological examinations showed a pattern of typical acute hepatitis (increase of transaminases and hepatocellular necnsis) in $CCl_4$-treated mice and cholestasis (increase of alkaline phosphatase and ${\gamma}$-glutamyltransferase, and biliary hyperplasia) in ANIT-treated mice, respectively, Mice were fasted at least 4 hr prior to the intravenous injection of $^{99m}$Tc-MAG3-biocytin (18.5 MBq/20$\mu\textrm{g}$) in 2% human serum albumin in saline. Scintigraphy was performed with a ${\gamma}$-camera equipped with a 1-mm diameter pin-hole collimator for 30 min and images were acquired every 15 s. We compared the values of physical parameters, such as peak liver/heart ratio ($${\gamma}$_{max}$) and peak ratio time ($t_{max}$) far $^{99m}$Tc-MAG3-biocytin scintigraphy. Results: Scintigraphic parameters of the $CCl_4$-pretreated (0.5 $m\ell$/kg) group showed a 81.9% decrease of r$_{max}$, and 42.2% decrease of $t_{max}$, whereas the ANIT-pretreated ( $150m\ell$/kg) group showed a 53% decrease of $r_{max}$, and 2.36-fold increase of $t_{max}$, (P<0.05). These results demonstrate that the decrease of $r_{max}$ and the shortening of $t_{max}$ are characteristic features for hepatotoxicity, in contrast to the increase of $t_{max}$ and decrease of $r_{max}$ for biliary hyperplasia. Conclusion: $^{99m}$Tc-MAG3-biocytin hepatobiliary scintigraphy can distinguish hepatitis from cholestasis in mice model and may be similarly useful in humans which differentiating the cause of neonatal jaundice in clinical study.cal study.cal study.cal study.

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Changes of Soil Properties and Evaluation of Plant Utilization According to the Application of Compost Used with Phyllite (Phyllite를 이용한 퇴비 시용에 따른 토양 특성 변화 및 작물에 대한 효용성 평가)

  • Park, Young-Hee;Chang, Ki-Woon;Hong, Jei-Gu
    • Journal of the Korea Organic Resources Recycling Association
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    • v.10 no.2
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    • pp.92-99
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    • 2002
  • The study was conducted to investigate the changes of soil properties and evaluation of plant utilization according to the application of compost used with phyllite. At this experiment, the plant cultivation test(lettuce and radish plant) was carried out to know about physico-chemical properties of compost, humus degradarion and stabilization degree, and some effects among rhe treatments were best at 20Mg/ha in PSPC 10(Phyllite, Saw dust, Pig manure Compost) plots and ar 10Mg/ha in PSPC 20 treatments, respectively. The EC value was high in the compost. Therefore the EC was appeared more than 2ds/m at the applied 40Mg/ha with compost. And in case of lettuce, Avail.-$P_2O_5$ and EC were more increased than in the radish plots, highly. In contrast, in case of radish, Toral-N, Total-C, and EC contents were decreased after plant cultivation. That is, it could know that radish was fond of absorprion of nitrogen. Also, BD values of soil physical properties were decreased according to increasing of phyllite addition ratio, however water holding capacity(WHC) was increased. The plant test had not been darnaged through the application of compost used with phyllite in soil and there had not been worse on changing of soil physico-chemical properties. Consequently, phyllite composts have been thought on agricultural utilized possibility.

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Determination of 3-phenoxybenzoic Acid in Urine and Exposure Assessment of Pyrethroid Insecticides to Human Being (요중 3-phenoxybenzoic acid 미량 분석 및 pyrethroid계 살포자 노출 평가)

  • Seo, Jong-Chul;Song, Jae-Seok;Choi, Hong-Soon
    • The Korean Journal of Pesticide Science
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    • v.11 no.2
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    • pp.87-94
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    • 2007
  • Pyrethroid insecticide have widely been used for agricultural sector and residential environments. To assess the exposure of insecticide which is absorbed through skin the analysis of urinary metabolite is essential. At present, the urinary 3-PBA was analyzed using liquid-phase extraction. But LPE have many limitations, such as long pre-treatment time and low recovery. So, this study was conducted to determine the optimum conditions for analysing 3-PBA in urine using solid phase extraction. Furthermore, this study intend to investigate the relation of concentrations of pyrethroid, deltamethrin in air and 3-PBA in urine. The optimum condition for hydrolysis was found to be done with hydrochloric acid for one hour. The recovery rates of 3-PBA were $84.6%{\pm}1.2%$, $54.8{\pm}0.9%$, $99.8{\pm}1.2%$ with XAD-2, XAD-7, XAD-16 using as the aborbents and acetone as eluents respectively. But acetonitrle and methanol gave low recovery rate and methyl cellosolve could not elute the compound. The amount of acetone for elution were 6mL, 9mL, 3mL for XAD-2, XAD-7, XAD-16 as absorbents respectively. The non-absorbed rates was $0.8{\pm}0.5%$, and $0.7{\pm}0.3%$ under XAD-16, mesh size 140-200, amount of resin 1.4g and the flow rate of eluent was 0.1mL/min. In the concentration process, we obtained 11 times higher concentration of material. The amounts of urinary 3-PBA were. The LODs of 3-PBA and deltamethrin were 0.004 mg/L, 0.038 mg/L, respectively. The further research of minute monitoring which include spray pattern, environmental condition is needed And more research about the relation between total pyrethroid exposure and urinary various metabolite are also necessary.

Partial Characterization of Physicochemical and Kinetic Properties of $Ca^{++}-ATPase$ System in Luteal Membranes (황체막에서의 $Ca^{++}-ATPase$의 특성)

  • Choi, Gyu-Bog;Koo, Bon-Sook;Kim, In-Kyo
    • The Korean Journal of Physiology
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    • v.20 no.2
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    • pp.257-270
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    • 1986
  • It has been reported that the luteal function may be regulated by the intracellular calcium in luteal cells (Higuchi et al, 1976; Dorflinger et at, 1984; Gore and Behrman, 1984) which is adjusted partially by $Ca^{++}-ATPase$ activities in luteal cell membranes (Verma and Pennistion, 1981). However, the physicochemical and kinetic properties of $Ca^{++}-ATPase$ in luteal membranes were not fully characterized. This study was, therefore, undertaken to partially characterize the physicochemical and kinetic properties of $Ca^{++}-ATPase$ system in luteal membranes and microsomal fractions, known as an one of the major $Ca^{++}$ storge sites (Moore and Pastan, 1978), from the highly luteinized ovary Highly luteinized ovaries were obtained from PMSG-hCG injected immautre female rats. Light membrane and heavy membrane fractions and microsomal fractions were prepared by the differential and discontinuous sucrose density gradient centrifugation method desribed by Bramley and Ryan (1980). Light membrane and heavy membrane fractions and microsomal fractions from highly luteinized ovaries are composed of the two different kinds of $Ca^{++}-ATPase$ system. One is the high affinity $Ca^{++}-ATPase$ which is activated in low $Ca^{++}$ concentration (Km, 10-30 nM), the other is low affinity $Ca^{++}-ATPase$ activated in higher $Ca^{++}$ concentration $(K_{1/2},\;40\;{\mu}M)$. At certain $Ca^{++}$ concentrations, activities of high and low affinity $Ca^{++}-ATPase$ are the highest in light membrane fractions and are the lowest in microsomal fractions. It appeares that high affinity $Ca^{++}-ATPase$ system have 2 binding sites for ATP (Hill's coefficient; around 2 in all membrane fractions measured) and the positive cooperativity of ATP bindings obviously existed in each membrane fractions. The optimum pH for high affinity $Ca^{++}-ATPase$ activation is around S in all membrane fractions measured. The lipid phase transition temperature measured by Arrhenius plots of high affinity $Ca^{++}-ATPase$ activity is around $25^{\circ}C$. The activation energies of high affinity $Ca^{++}-ATPase$ below the transition temperature are similar in each membrane fractions, but at the above transition temperature, it is the hightest in heavy membrane fractions and the lowest in microsomal fractions. According to the above results, it is suggested that intracellular $Ca^{++}$ level, which may regulate the luteal function, may be adjusted primarily by the high affinity $Ca^{++}-ATPase$ system activated in intracellular $Ca^{++}$ concentration range $(below\;0.1\;{\mu}M)$.

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Developmemt of Rice Husk Pellets as Bio-filter Media of Bio Scrubber Odor Removal System (왕겨펠렛 생물담체 개발 및 이를 이용한 bio scrubber형 악취제거 시스템 성능평가)

  • Bae, Jiyeol;Han, Sangjong;Park, Ki Ho;Kim, Kwang-Soo
    • Journal of Korean Society for Atmospheric Environment
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    • v.34 no.4
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    • pp.554-566
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    • 2018
  • The rice husk contains nutrients which can be easily utilized by microorganisms, and also has a water retaining ability, which played a crucial part in enabling it to become a biofilter media. In this study, we evaluated the applicability of rice husk pellet bio-scrubber as a microbiological carrier. The pelletization experiment of rice husk as a biological media was performed using PVA and EVA binder. Also, the feasibility tests of rice husk as a biological media for odor removal were carried out in order to know whether rice-husk contains useful components as a media for microbiological growth or not. Lastly, a combined test for odor gas absorption and biological oxidation was conducted using a lab scale bio-filter set-up packed with rice-husk pellets as wet-scrubber. The major components of the rice husk were carbon, hydrogen, nitrogen, and oxygen, while carbon acted as the main ingredient which comprised up to 23.00%. The C : N : P ratio was calculated as 45 : 1 : 2. Oxygen uptake rate, yield and decay rate of the rice husk eluent was calculated to be $0.0049mgO_2/L/sec$, 0.24 mgSS/mgCOD and 0.004 respectively. The most stable form of rice husk pellets was produced when the weight of the rice husk, EVAc, PVAc, and distilled water was 10 : 2 : 0.2 : 10. The prepared rice husk pellets had an apparent density of 368 g/L and a porosity of 59.00% upon filling. Dry rice husks showed high adsorption capacity for ammonia gas but low adsorption capacity for hydrogen sulfide. The bio-filter odor removal column filled with rice husk pellets showed more than 99.50% removal efficiency for NH3 and H2S gas. Through the analysis of circulation water, the prime removal mechanism is assumed to be the dissolution by water, microbial nitrification, and sulfation. Finally, it was confirmed that the microorganisms could survive well on the rice husk pellets, which provided them a stable supply of nutrients for their activity in this long-term experiment. This adequate supply of nutrients from the rice husk enabled high removal efficiency by the microorganisms.

Comparison of Opened Rates and Quality Characteristics of Frozen Baby-clam In-shell Tapes philippinarum Prepared by Different Processing Method (제조방법을 달리하여 제조한 껍질붙은 냉동바지락(Tapes philippinarum)의 껍질 개패율 및 품질특성 비교)

  • Park, Si-Young;Kang, Kyung-Hun;Lee, Jae-Dong;Yoon, Moon-Joo;Kang, Young-Mi;Seoung, Tae-Jong;Kweon, Su-Hyun;Choo, Yi-Kwon;Kim, Jeong-Gyun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.6
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    • pp.743-749
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    • 2016
  • We compared two different processing methods for preparing high quality frozen in-shell baby clam products. In the first method, sand and mud were removed from the clams, then they were vacuum packed in polyethylene film, boiled at $97^{\circ}C$ for 6 min, and snap frozen in a cold air blast freezer (sample 1). The second processing method was similar, except the boiling process was excluded (sample 2). Both frozen products were boiled for 4 min, and then shucked and minced. Various quality metrics, such as the opening rates of shells, chemical composition, pH, volatile basic nitrogen (VBN), salinity, thiobarbituric acid (TBA), amino-N, total amino acids and free amino acids were measured, and sensory evaluation was conducted. The opening rates of shells of sample 1 and sample 2 were 98.3% and 4.67%, respectively. The proximate composition of sample 1 and sample 2 was 75.2% and 78.7% moisture, 19.7% and 16.2% crude protein, 2.45 and 2.2% crude lipid, 2.8% and 2.1% ash, and 2.1% and 1.9% salinity, respectively. The L, a, b and ${\Delta}E$ values were similar: 48.6 and 49.2, 3.9 and 3.9, 15.7 and 15.5, and 50.7 and 50.1 for sample 1 and sample 2, respectively. The sensory evaluation score of sample 1 was higher than that of sample 2. Sample 1 was deemed to be superior to sample 2; therefore, we determined that the boiling process is needed for manufacturing high-quality frozen clam products.

Analysis and Comparison of Cerebroside Components from Soybean Fermented Foods (대두발효식품 Cerebroside의 구성성분 분석 및 비교)

  • 이은열;김희숙
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.2
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    • pp.177-183
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    • 2002
  • Cerebroside fatty acids, sugars and long-chain sphingoid bases in raw soybean and soybean fermented foods (chongkukjang and deunjang) were analyzed using gas chromatography-mass spectrometry (GC-MS) and high-pH anion exchange chromatography with pulsed amerometric detection (HPAEC-PAD). Fatty acids of acid-hydrolyzed cerebrosides were derivatized to O-TMS methylester and analysed. The major fatty acids in raw soybean and chongkukjang cerebrosides were identified as 2-hydroxyhexadecanoic acid (16 : 0h), 2-hydroxydocosanoic acid (22 : 0h) and 2-hydroxytetracosanoic acid (24 : 0h). In the case of deunjang cerebroside, 24 : 0h (40.9%) and 22 : 0h (23.4%) were major fatty acids, but 16 : 0h, 23 : 0h, 25 : 0h and 26 : 0h were also detected. Long-chain sphingoid bases of acid-hydrolyzed cerebrosides from raw soybean, chongkukjang and deunjang consisted primarily of 4-tracts, 8-tracts-sphingadienine (dihydroxy base, d18 : 2$\Delta$$^{4trans, 8trans}$) and sis-tracts isomers of 4-hydroxy-sphingenine (trihydroxy base, tl8:1$\Delta$$^{4trans or cis}$) with much less amounts of phytosphingosine (tl8: 0) and isomers of sphingenine (d18 : 1). Although deunjang is a soybean food fermented by fungi and microorganisms for a long period, 2-hydroxyoctadec-3-enoic acid (18 : 1h) and branched 9-methyl-4,8-sphingadienine known as compositional cerebroside fatty acids in Aspergillus species were not detected. Mass spectrum for sugar derivatives in cerebrosides of soybean foods including raw soybean and fermented soybean showed that C-1 of glucose moiety was linked to ceramide backbone as like a monoglucosylceramide.

Effects of Korean Red Ginseng (Panax ginseng), urushiol (Rhus vernicifera Stokes), and probiotics (Lactobacillus rhamnosus R0011 and Lactobacillus acidophilus R0052) on the gut-liver axis of alcoholic liver disease

  • Bang, Chang Seok;Hong, So Hyung;Suk, Ki Tae;Kim, Jin Bong;Han, Sang Hak;Sung, Hotaik;Kim, Eun Ji;Kim, Myoung Jo;Kim, Moon Young;Baik, Soon Koo;Kim, Dong Joon
    • Journal of Ginseng Research
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    • v.38 no.3
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    • pp.167-172
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    • 2014
  • Background: Roles of immune reaction and toll-like receptor-4 (TLR-4) have widely been established in the pathogenesis of alcoholic liver disease (ALD). Methods: We evaluated the biologic efficacy of Korean Red Ginseng (KRG), urushiol, and probiotics (Lactobacillus rhamnosus R0011 and Lactobacillus acidophilus R0052) in mouse models of ALD. Sixty C57BL/6 mice were equally divided into six feeding groups for 10 weeks: normal diet, alcohol, control, alcohol + KRG, alcohol + urushiol, and alcohol + probiotics. Alcohol was administered via a LiebereDeCarli liquid diet containing 10% alcohol. TLR-4 expression, proinflammatory cytokines, and histology, as well as the results of liver function tests were evaluated and compared. Results: No between-group differences were observed with regard to liver function. TLR-4 levels were significantly lower in the KRG, urushiol, and probiotics groups than in the alcohol group ($0.37{\pm}0.06ng/mL$, $0.39{\pm}0.12ng/mL$, and $0.33{\pm}0.07ng/mL$, respectively, vs. $0.88{\pm}0.31ng/mL$; p < 0.05). Interleukin-$1{\beta}$ levels in liver tissues were decreased among the probiotics and KRG groups. The tumor necrosis factor-${\alpha}$ level of liver tissue was decreased in the KRG group. Conclusion: The pathological findings showed that alcohol-induced steatosis was significantly reduced by KRG and urushiol. As these agents improve immunologic capacity, they may be considered in potential anti-ALD treatments.

In vivo Pharmacokinetics, Activation of MAPK Signaling and Induction of Phase II/III Drug Metabolizing Enzymes/Transporters by Cancer Chemopreventive Compound BHA in the Mice

  • Hu, Rong;Shen, Guoxiang;Yerramilli, Usha Rao;Lin, Wen;Xu, Changjiang;Nair, Sujit;Kong, Ah-Ng Tony
    • Archives of Pharmacal Research
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    • v.29 no.10
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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.