• Asian-Australasian Journal of Animal Sciences
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• v.22 no.2
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• pp.168-173
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• 2009

Human follicle-stimulating hormone (hFSH) is a pituitary glycoprotein that regulates follicular development and ovulation. Clinically, hFSH has been used to induce follicular growth in infertile women. The hormone is composed of heterodimers, including a common ${\alpha}$ subunit among the gonadotropin family and a hormone-specific ${\beta}$ subunit. Since assembly of the heterodimer is a rate-limiting step in the production of functional hFSH, transgenic clone cows carrying a single-chain hFSH transgene may efficiently produce functional hormone. Genes encoding the ${\alpha}$ and ${\beta}$ subunits of hFSH were linked using the C-terminal peptide sequence from the ${\beta}$ subunit of human chorionic gonadotropin. Bovine fetal fibroblasts were transfected with the gene construct, including the goat ${\beta}$-casein promoter and a single-chain hFSH coding sequence. Transfected fibroblasts were transferred into enucleated oocytes, and individual nuclear transfer (NT) embryos developed to the blastocyst stage were analyzed for the transgene by polymerase chain reaction. Seventy eight blastocysts (30.8%) were developed from 259 reconstructed embryos. Among these blastocysts, the hFSH gene was detected in 70.8% (34/48) of the embryos. Subsequent transfer of hFSH-transgenic clone embryos to 31 recipients results in 11 (35.5%) early pregnancies. However, all fetuses were lost before reaching day 180 of gestation. The results from this study demonstrated that bovine NT embryos carrying single-chain hFSH could be produced, and further extensive studies in which NT embryos are transferred to more recipients may give rise to single chain hFSH-transgenic cows for biomedical applications.

• Journal of Embryo Transfer
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• v.9 no.3
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• pp.229-234
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• 1994

This experiment was carried out to develop the model system for mass production of biomedical and nutritional proteins (human proteins) through mamraary gland of the transgenic cattle produced by gene manipulation and embryological technologies. Human growth hormone gene fused with rat $\beta$-casein gene promoter was microinjected into pronuclei of one cell bovine embryos produced by in vitro fertilization. After microinjection, embryos were cultured in vitro for 6 or 7 days. Twenty embryos reaching to blastocysts were transferred to 10 beef recipients, each receiving two embryos. Recipients were diagnosed for pregnancy by rectal palpation at 76 days after embryo transfer. One of them was pregnant to term and produced a female calf weighing 21 kg at 280 days following embryo transfer. DNA was extracted from umbilical cord tissue and blood of calf born for confirming gene insertion. As determined by Southern hybridization, the transgene was not found.

• Korean Journal of Microbiology
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• v.16 no.4
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• pp.141-147
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• 1978

In order to increase the productivity of cellulolytic enzymes, medium composition and culture conditions were studied. When cellulose powder (Avicel) supplemented with rice straw was used as carbon source, productivity of ${\beta}-glucosidase$ was increased by about 3 times compared with the runs with only cellulose powder as a carbon source. In this case no negative effects on the production of CMC enzyme activity and filter paper activity was found. For the production of celulolytic enzymes using T. reesei QM 9414, casitone was found to be a good nitrogen source compared with other sources studied, such as peptone, yeast extract, tryptone, and casein. The highest cellulase activity was attained when 0.3% glucose and 0.01% Tween 80 were supplemented to the standard medium of Rese. An adequate oxygen transfer rate was also found to be important to the cellulase fermentation and about 50 mmole of oxygen/liter/hour supported good cellulase biosynthesis during cellulase fermentation.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.11
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• pp.1634-1641
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• 2005

Generally, dietary fibre (DF) includes lignin, non-starch polysaccharides (NSP) and resistant starch (RS). In monogastric species, low levels of dietary fibre in the diet are associated with various diseases and high levels reduce nutrient digestibilities. In this study, the effects of different types and levels of NSP (soluble: $\beta$-glucan, insoluble cellulose) and resistant starch on mucin secretion and endogenous nitrogen and amino acid losses in pigs were investigated. A total of 25 five-week-old weaner pigs (9.5 kg${\pm}$1.5 kg), were randomly allocated to each of five experimental diets. Different levels of purified barley $\beta$-glucan (BG) extract (5 or 10% of $Glucagel^{(R)}$ $\beta$-glucan, providing 4 or 8% of BG in the diet), and resistant starch (RS) (8.3 or 16.6% of Hi-$Maize^{TM}$, providing 5 or 10% RS in the diet) were substituted for wheat starch in a purified diet in which enzymatically-hydrolysed casein was the sole source of protein. The diets were fed for 21 days. No statistically significant difference between treatments (p>0.05) was observed for growth performance and organs weights. No difference in ileal starch digestibility was observed between pigs on the cellulose or $\beta$-glucan diets. However, as the level of resistant starch in the diet increased the ileal starch digestibility decreased (p<0.05). The inclusion of resistant starch in the diet (5 or 10%) did not increase mucin production when compared with the cellulose-only diet. However, as the level of beta-glucan in the diet increased, both crude mucin in the digesta dry matter and per kg dry matter intake increased (p<0.05). Pigs fed the diet containing 8% of beta-glucan had higher endogenous loss flow than those fed the diets including 5 or 10% of resistant starch or 4% of $\beta$-glucan. In conclusion, dietary inclusion of resistant starch increased the level of starch reaching the large intestine without any effect on mucin secretion, or endogenous nitrogen or amino acid losses content in the small intestine. The addition of $\beta$-glucan to a diet containing cellulose increases both mucin secretion and endogenous amino acid and nitrogen losses in the small intestine.

• Biomolecules & Therapeutics
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• v.26 no.3
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• pp.268-273
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• 2018

Sleep is the most basic and essential physiological requirement for mental health, and sleep disorders pose potential risks of metabolic and neurodegenerative diseases. Tryptic hydrolysate of ${\alpha}_{S1}$-casein (${\alpha}_{S1}-CH$) has been shown to possess stress relieving and sleep promoting effects. However, the differential effects of ${\alpha}_{S1}-CH$ on electroencephalographic wave patterns and its effects on the protein levels of ${\gamma}$-aminobutyric acid A ($GABA_A$) receptor subtypes in hypothalamic neurons are not well understood. We found ${\alpha}_{S1}-CH$ (120, 240 mg/kg) increased sleep duration in mice and reduced sleep-wake cycle numbers in rats. While ${\alpha}_{S1}-CH$ (300 mg/kg) increased total sleeping time in rats, it significantly decreased wakefulness. In addition, electroencephalographic theta (${\theta}$) power densities were increased whereas alpha (${\alpha}$) power densities were decreased by ${\alpha}_{S1}-CH$ (300 mg/kg) during sleep-wake cycles. Furthermore, protein expressions of $GABA_A$ receptor ${\beta}_1$ subtypes were elevated in rat hypothalamus by ${\alpha}_{S1}-CH$. These results suggest ${\alpha}_{S1}-CH$, through $GABA_A$ receptor modulation, might be useful for treating sleep disorders.

• The Korean Journal of Physiology and Pharmacology
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• v.21 no.2
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• pp.197-204
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• 2017

In the present study, we tried to examine whether oleanolic acid regulates the activity, secretion and gene expression of matrix metalloproteinase-3 (MMP-3) in primary cultured rabbit articular chondrocytes, as well as the production of MMP-3 in the knee joint of rat to evaluate the potential chondroprotective effect of oleanolic acid. Rabbit articular chondrocytes were cultured in a monolayer, and reverse transcription-polymerase chain reaction (RT-PCR) was used to measure interleukin-$1{\beta}$ (IL-$1{\beta}$)-induced gene expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), ADAMTS-5 and type II collagen. In rabbit articular chondrocytes, the effects of oleanolic acid on IL-$1{\beta}$-induced secretion and proteolytic activity of MMP-3 were investigated using western blot analysis and casein zymography, respectively. The effect of oleanolic acid on in vivo MMP-3 protein production was also examined, after intra-articular injection to the knee joint of rat. The results were as follows: (1) oleanolic acid inhibited the gene expression of MMP-3, MMP-1, MMP-13, ADAMTS-4, and ADAMTS-5, but increased the gene expression of type II collagen; (2) oleanolic acid reduced the secretion and proteolytic activity of MMP-3; (3) oleanolic acid suppressed the production of MMP-3 protein in vivo. These results suggest that oleanolic acid can regulate the activity, secretion and gene expression of MMP-3, by directly acting on articular chondrocytes.

• The Korean Journal of Physiology and Pharmacology
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• v.20 no.2
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• pp.221-228
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• 2016

We investigated whether prunetin affects the proteolytic activity, secretion, and gene expression of matrix metalloproteinase-3 (MMP-3) in primary cultured rabbit articular chondrocytes, as well as in vivo production of MMP-3 in the rat knee joint to evaluate the potential chondroprotective effect of prunetin. Rabbit articular chondrocytes were cultured in a monolayer, and reverse transcriptionpolymerase chain reaction (RT-PCR) was used to measure interleukin-$1{\beta}$ (IL-$1{\beta}$)-induced expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), and ADAMTS-5. In rabbit articular chondrocytes, the effects of prunetin on IL-$1{\beta}$-induced secretion and proteolytic activity of MMP-3 were investigated using western blot analysis and casein zymography, respectively. The effect of prunetin on MMP-3 protein production was also examined in vivo. The results were as follows: (1) prunetin inhibited the gene expression of MMP-3, MMP-1, MMP-13, ADAMTS-4, and ADAMTS-5; (2) prunetin inhibited the secretion and proteolytic activity of MMP-3; (3) prunetin suppressed the production of MMP-3 protein in vivo. These results suggest that prunetin can regulate the gene expression, secretion, and proteolytic activity of MMP-3, by directly acting on articular chondrocytes.

• Biomolecules & Therapeutics
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• v.24 no.2
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• pp.163-170
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• 2016

We examined whether apigenin affects the gene expression, secretion and activity of matrix metalloproteinase-3 (MMP-3) in primary cultured rabbit articular chondrocytes, as well as in vivo production of MMP-3 in the knee joint of rat to evaluate the potential chondroprotective effects of apigenin. Rabbit articular chondrocytes were cultured in a monolayer, and reverse transcription - polymerase chain reaction (RT-PCR) was used to measure interleukin-$1{\beta}$ (IL-$1{\beta}$)-induced expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), and ADAMTS-5. In rabbit articular chondrocytes, the effects of apigenin on IL-$1{\beta}$-induced secretion and proteolytic activity of MMP-3 were investigated using western blot analysis and casein zymography, respectively. The effect of apigenin on MMP-3 protein production was also examined in vivo. In rabbit articular chondrocytes, apigenin inhibited the gene expression of MMP-3, MMP-1, MMP-13, ADAMTS-4, and ADAMTS-5. Furthermore, apigenin inhibited the secretion and proteolytic activity of MMP-3 in vitro, and inhibited production of MMP-3 protein in vivo. These results suggest that apigenin can regulate the gene expression, secretion, and activity of MMP-3, by directly acting on articular chondrocytes.

• Biomolecules & Therapeutics
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• v.22 no.3
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• pp.239-245
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• 2014

We investigated whether luteolin affects the gene expression, secretion and activity of matrix metalloproteinase-3 (MMP-3) in primary cultured rabbit articular chondrocytes, as well as production of MMP-3 in the rat knee to evaluate the potential chondroprotective effects of luteolin. Rabbit articular chondrocytes were cultured in a monolayer and IL-$1{\beta}$-induced gene expression levels of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), ADAMTS-5 and type II collagen were measured by reverse transcription - polymerase chain reaction (RT-PCR). Effects of luteolin on interleukin- $1{\beta}$ (IL-$1{\beta}$)-induced secretion and enzyme activity of MMP-3 in rabbit articular chondrocytes were investigated by western blot analysis and casein zymography, respectively. The effect of luteolin on MMP-3 protein production was also examined in vivo. The results were as follows: (1) luteolin inhibited the gene expression levels of MMP-3, MMP-1, MMP-13, ADAMTS-4 and ADAMTS-5. However, it increased the gene expression level of collagen in rabbit articular chondrocytes; (2) luteolin inhibited the secretion and activity of MMP-3; (3) luteolin inhibited in vivo production of MMP-3 protein. These results suggest that luteolin can regulate the gene expression, secretion and activity of MMP-3, by directly acting on articular chondrocytes.

• Plant Biotechnology Reports
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• v.3 no.3
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• pp.243-250
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• 2009

In the present study, a simple one medium formulation protocol for callus culture, somatic embryogenesis and in vitro production of ${\beta}-carboline$ alkaloids and diosgenin in Tribulus terrestris L. was developed. Extensive callus induction and proliferation was obtained in leaf explant on Murashige and Skoog (MS) medium supplemented with $5.0{\mu}M$ 6 benzyl adenine (BA) and $2.5{\mu}M$ ${\alpha}-naphthaleneacetic$ acid (NAA). The embryogenic callus was maintained on subculture to fresh parental medium at 4-week intervals over a period of 28 months. The frequency of embryo formation was at a maximum ($18.1{\pm}0.9$ per g of callus) on MS medium containing $5.0{\mu}M$ BA and $2.5{\mu}M$ NAA together with $75mg\;1^{-1}$ casein hydrolysate. Globular embryo developed into torpedo stage embryo under the influence of starvation. The accumulation of ${\beta}-carboline$ alkaloids (harmaline and harmine) and steroidal saponin (diosgenin) in non-embryogenic and embryogenic callus culture derived from leaf explant was compared with root, leaf, stem, and fruit of the mother plant. The embryogenic callus accumulated equivalent amounts of harmaline ($66.4{\pm}0.5{\mu}g/g$ dry weight), harmine ($82.7{\pm}0.6{\mu}g/g$ dry weight), and diosgenin ($170.7{\pm}1.0{\mu}g/g$ dry weight) to that of the fruit of T. terrestris. The embryogenic callus culture of this species might offer a potential source for production of important pharmaceuticals.