• Clinical and Experimental Pediatrics
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• v.50 no.6
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• pp.576-579
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• 2007

The syndrome of resistance to thyroid hormone (RTH) is characterized by reduced tissue sensitivity to thyroid hormone (TH). In the majority of subjects, RTH is caused by mutations in the thyroid hormone receptor beta ($TR{\beta}$) gene, located on the chromosome locus 3p24.3. RTH is inherited in an autosomal dominant manner. The clinical presentation of RTH is variable, but common features include elevated serum levels of thyroid hormone (TH), a normal or slightly increased thyrotropin (thyroid stimulating hormone, TSH) level that responds to thyrotropin releasing hormone (TRH), and goiter. We report a 4 year-old girl, who was clinically euthyroid in spite of high total and free $T_4$, and $T_3$ concentrations, while TSH was slightly increased. Sequence analysis of the thyroid hormone receptor beta gene (THRB) confirmed a heterozygous C to T change at nucleotide number 1303, resulting in a substitution of histidine by tyrosine at codon 435 (H435Y). Further analysis of her parents revealed that the H435Y variation was a de novo mutation since neither parents had the variation. Her parents' TH and TSH levels were within normal range.

• Journal of Microbiology and Biotechnology
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• v.19 no.12
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• pp.1650-1655
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• 2009

The $\beta$-glucuronidase (GUS) gene from Lactobacillus brevis RO1 was cloned and expressed in Escherichia coli GMS407. The GUS gene was composed of 1,812 bp, encoding a 603-amino-acid protein belonging to glycosyl hydrolase family 2 with three conserved domains. The amino acid similarity was higher than 70% with the $\beta$-glucuronidases of various microorganisms, yet less than 58% with the $\beta$-glucuronidase of L. gasseri ADH. Overexpression and purification of the GUS was performed in $\beta$-glucuronidase-deficient E. coli GMS407. The purified GUS protein was 71 kDa and showed 1,284 U/mg of specific activity at optimum conditions of pH 5.0 and $37^{\circ}C$. At $37^{\circ}C$, the GUS remained stable for 80 min at pH values ranging from 5.0 to 8.0. The purified enzyme exhibited a half-life of 1 h at $60^{\circ}C$ and more than 2 h at $50^{\circ}C$. When the purified GUS was applied to transform baicalin and wogonoside into their corresponding aglycones, $150\;{\mu}M$ of baicalin and $125\;{\mu}M$ of wogonoside were completely transformed into baicalein and wogonin, respectively, within 3 h.

• Membrane Journal
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• v.13 no.4
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• pp.291-299
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• 2003

Polymer membranes such as poly(1-trimethylsilyl-1-propyne)-polyetherimide (PTMSP-PEI) and poly(dimethylsiloxane)- polyetherimide (PDMS-PEI) composite membrane were prepared by solution casting method. To investigate the characteristics of these membranes, the analytical methods such as FT-IR, $^1H-NMR,$ DSC, TGA, GPC, and SEM have been utilized. The number-average (equation omitted) and weight-average (equation omitted) molecular weight of PTMSP were 477,920 and 673,329 respectively. The glass transition temperature ($T_g$) of PTMSP was $224^{\circ}C.$ The separation of the gas mixture ($H_2/N_2$) through the composite membranes were studied as a function of pressure. The separation factor (${\alpha}, {\beta},$ quation omitted) of the composite membranes used in this work increased as the pressure of permeation cell increased. The real separation factor (${\alpha}$), head separation factor (${\beta}$), and tail separation factor (equation omitted) of PTMSP-PEI composite membrane were 2.28, 1.17, and 1.96 respectively at ${\Delta}P$ 30psi and $25^{\circ}C.$ (${\alpha}, {\beta}$ and equation omitted of PDMS-PEI composite membrane were 3.70, 1.53, and 2.42 respectively at ${\Delta}P$ 30psi and $25^{\circ}C$.

• Biomolecules & Therapeutics
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• v.25 no.6
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• pp.593-598
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• 2017

The $Na^+/H^+$ exchanger-1 (NHE-1) is a ubiquitously expressed pH-regulatory membrane protein that functions in the brain, heart, and other organs. It is increased by intracellular acidosis through the interaction of intracellular $H^+$ with an allosteric modifier site in the transport domain. In the previous study, we reported that glutamate-induced NHE-1 phosphorylation mediated by activation of protein kinase C-${\beta}$ (PKC-${\beta}$) in cultured neuron cells via extracellular signal-regulated kinases (ERK)/p90 ribosomal s6 kinases (p90RSK) pathway results in NHE-1 activation. However, whether glutamate stimulates NHE-1 activity solely by the allosteric mechanism remains elusive. Cultured primary cortical neuronal cells were subjected to intracellular acidosis by exposure to $100{\mu}M$ glutamate or 20 mM $NH_4Cl$. After the desired duration of intracellular acidosis, the phosphorylation and activation of PKC-${\beta}$, ERK1/2 and p90RSK were determined by Western blotting. We investigated whether the duration of intracellular acidosis is controlled by glutamate exposure time. The NHE-1 activation increased while intracellular acidosis sustained for >3 min. To determine if sustained intracellular acidosis induced NHE-1 phosphorylation, we examined phosphorylation of NHE-1 induced by intracellular acidosis by transient exposure to $NH_4Cl$. Sustained intracellular acidosis led to activation and phosphorylation of NHE-1. In addition, sustained intracellular acidosis also activated the PKC-${\beta}$, ERK1/2, and p90RSK in neuronal cells. We conclude that glutamate stimulates NHE-1 activity through sustained intracellular acidosis, which mediates NHE-1 phosphorylation regulated by PKC-${\beta}$/ERK1/2/p90RSK pathway in neuronal cells.

• KSBB Journal
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• v.16 no.2
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• pp.174-178
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• 2001

Soybeans contain the phytoestrogens genistein and daidzein, their glucosides genistin and daidzin and coumesterol. These isoflavonoid compounds are capable of producing an estrogenic response in a number of diverse species. This study determined optimum conditions for the extraction of the main isoflavones(daidzin, genistin, daidzein, genistein) in defatted soybean meal using high-performance liquid chromatography. The best optimum extraction was achieved at 75% ethanol, $80^{circ}C$, pH4 and a three hour contact time. In addition, isoflavones with high purity were separated by adding up to 4%(w/v) of calcium chloride dihydrate. Most soybean extracts were composed of $beta$-glucosidic conjugate(daidzin, genistin) which is difficult to adsorb in body. Therefore, $beta$-glucosidase was used to convert as conjugate to aglycone form (daidzein, genistein) which is easy to adsorb. The optimal conditions of enzyme reaction involved to be 8.4 units of enzyme concentration, pH5.0, $40^{circ}C$ and 40 minutes.

• Molecules and Cells
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• v.25 no.2
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• pp.312-316
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• 2008

Flavanone $3{\beta}$-hydroxylases (F3H) are key enzymes in the synthesis of flavonol and anthocyanin. In this study, three F3H cDNAs from Oryza sativa (OsF3H-1 ~3) were cloned by RT-PCR and expressed in E. coli as gluthatione S-transferase (GST) fusion proteins. The purified recombinant OsF3Hs used flavanone, naringenin and eriodictyol as substrates. The reaction products with naringen and eriodictyol were determined by nuclear magnetic resonance spectroscopy to be dihydrokaempferol and taxifolin, respectively. OsF3H-1 had the highest enzymatic activity whereas the overall expression of OsF3H-2 was highest in all tissues except seeds. Flavanone $3{\beta}$-hydroxylase could be a useful target for flavonoid metabolic engineering in rice.

• Korean Journal of Animal Reproduction
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• v.17 no.4
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• pp.375-383
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• 1994

The human growth hormone (hGH) gene uder the control of the rat $\beta$-casein promoter gene was designed to produce transgenic mouse expressed hGH gene in only mammary gland. One hundred seventy two eggs microinjected were transferred to the oviducts of pseudopregnants and 43 offspring were delivered. By Southern blotting hybridization, 3 were transgenic with rat $\beta$-casein/hGH gene. The copy numbers of three transgenic founder were 1, 5, and 15, respectively. A radioimmunoassay was developed to quantitate the amount of expression of the hGH gene in mammary gland of transgenic mice. The amount of hGH was 13.3ng/ml in the lactating milk of one transgenic line, showing predominantly higher than 3.0ng/ml in milk of control mice. Therefore, our findings suggested that $\beta$-casein promoter may induce the tissue specific expression of structural gene.

• Food Science of Animal Resources
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• v.38 no.4
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• pp.780-793
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• 2018

This study was conducted to compare the anti-allergic effects of a whey protein concentrate (WPC) and WPC hydrolysate. WPC hydrolysate was prepared using enzymatic digestion for 8 h with trypsin and ${\alpha}$-chymotrypsin, after which it was freeze-dried. The allergic parameters assessed in rat basophilic leukemia (RBL)-2H3 cells were degranulation and release of ${\beta}$-hexosaminidase, release of tumor necrosis factor $(TNF)-{\alpha}$, and changes in the expression of $IL-1{\beta}$, IL-4, and IL-10 by real time polymerase chain reaction (PCR). During preparation of the WPC hydrolysate, hydrolysis increased rapidly from 0 to 10 min and then gradually increased slowly from 1 h onwards, achieving a final degree of hydrolysis of 78.50%. The SDS-PAGE analysis revealed a reduction in the intensity of several protein bands in the WPC hydrolysate compared to the WPC. IgE-induced ${\beta}$-hexosaminidase release from RBL-2H3 cells was decreased to a higher degree following treatment with the hydrolysate compared to WPC treatment. W500 ($500{\mu}g/mL$ WPC) showed the least inhibition of ${\beta}$-hexosaminidase release, but there was no significant difference between W500 and W1000 ($1,000{\mu}g/mL$) (p<0.05). H1000 ($1,000{\mu}g/mL$ WPC hydrolysate) inhibited ${\beta}$-hexosaminidase release by 39%. Compared to the control, treatment with H1000 decreased $TNF-{\alpha}$ secretion to 11.87 pg/mL. The gene expression levels of IL-1${\beta}$, IL-4, and IL-13 were all significantly decreased in hydrolysate (p<0.05). In the case of $IL-1{\beta}$ and IL-4, the expression levels in W1000 treated cells were decreased by 73.67% and 65%, respectively, and that of IL-13 was decreased by 66.43% compared to the control.

• Korean Journal of Food Science and Technology
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• v.36 no.3
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• pp.440-447
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• 2004

Characteristics of human ${\beta}-carotene$ 15,15'-dioxygenase isolated by recombinant DNA technology was elucidated. Optimal pH and temperature were 9.0 and $40^{\circ}C$, respectively. Enzyme activity was temperature-sensitive. Enzyme was stable at pH 6.0-9.0 for 24 hr and under $5^{\circ}C$. Half-life of enzyme at $35^{\circ}C$ was 40 min. Crude preparations of enzyme were inhibited by ferrous ion-chelating agent and sulfhydryl-binding agent. GSH offsets inhibitory effect of PCMB. With increasing substrate concentrations, enzyme activity gave typical Michaelis-Menten curve, Based on Hanes-Woolf plot of data, $K_{m}\;and\;V_{max$ were $3.39{\times}10^{6}\;M\;and\;1.2\;pmol/mg$ protein/min, respectively.

• Microbiology and Biotechnology Letters
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• v.30 no.2
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• pp.151-156
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• 2002

This study was performed to obtain the thermostable $\beta$-glycosidase producing bacteria from hot spring of volcanic area at Atagawa in Japan. KNOUC 202 was selected because it showed thermostable $\beta$-glycosidase activity in sodium phosphate buffer(pH 6.8) at $70^{\circ}C$ for 4h, and it was identified. The strain was aerobic, asporogenic bacilli, immobile, gram negative, catalase positive, oxidase positive, and pigment-producing. Optimum growth was at $70~72^{\circ}C$, pH 7.0~7.2, and it could grow in the presence of 3% NaCl. The main fatty acids in cell were iso-15:0 and iso-l7:0. 16S rRNA sequence of KNOUC 202 showed 99.9% similarity with that of Thermus thermophilus ATCC 27634(HB8). Based on morphological, physiological, biochemical characteristics, cellular fatty acids profile and 16S rRNA sequence analysis, KNOUC 202 was identified as Thermus thermophilus.