• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.6
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• pp.371-374
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• 2002

Production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate)［P(3HB/V)］, by fed-batch culture of recombinant Escherichia coli harboring a plasmid containing the Alcaligenes latus polyhy-droxyalkanoate (PHA) biosynthesis genes, was examined in two pilot-scale fermentors with air supply only, In a 30 L fermentor having a XLa value of 0.11 S­$^1$, the final P(3HB/V) concentration and the P(3HB/V) content obtained were 29.6 g/L and 70.1 wt%, respectively giving a productivity of 1.37 g P(3HB/V)/L-h. In a 300 L fermentor having a XLa of 0.03 S­$^1$, the P(3HB/V) concentration and the P(3HB/V) content were 20.4 g/L and 69 wt%, respectively giving a productivity of 1.06g P(3HB/V)/L-h. These results suggest that economical production of P(3HB/V) is possible by fed-batch culture of recombinant E. coli in a large-scale fermentor having low KLa value.

• KSBB Journal
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• v.14 no.4
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• pp.422-428
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• 1999

A transformat Alcaligence eutrophus GA5 harboring phbC gene from A. latus was cultivated for production of Poly(3-hydroxybutyrate-4-hydroxybutyrate)[P(3HB-4HB)] containing high molar fraction of 4-hydroxybutyrate(4HB)] containing high molar fraction of 4-hydroxybutyrate(4HB). Transformation did not influenced significantly on total cell growth, on total cell growth, concentration, and content of P(3HB-4HB), however, significantly influenced on 4HB molar fraction in P(3HB-4HB) increasing from 12.3 to 23.5 mol% after 48 h cultivation in two-stage using 1.0%(W/V) of ${\gamma}$-butyrolactone as a precursor compare to parent strain. Above increment may be due to the accelerated polymerization between 3HB and 4HB converted from precusor compound by amplified phbC gene. Citrate increased remarkbly total cell mass and P(3HB-4HB) concentration, but did not influenced on the molar fraction of 4HB, meanwhile, magnesium ion influenced on P(3HB-4HB) concentration and 4HB molar fraction significantly. The two-stage cultivation method was modified, in such a way minimizing P(3HB) accumulated inside of cell grown at first-stage, consquently, 26.3% of P(3HB-4HB) containing 61.0 mol% of 4HB fraction was obtained after 72hr. Furthermore, semi-homopolymeric P(4HB) containing 92.0 mol% of 4Hb was obtained, and its structure was confirmed by $^1$H-NMR.

• Journal of Embryo Transfer
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• v.29 no.2
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• pp.127-132
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• 2014

This study was carried out to investigate the effects of tissue inhibitor of matalloproteinase-1 (TIMP-1), Activin A and Heparin binding epidermal growth factor (HB-EGF) on in vitro production of bovine embryos. In experiment 1, presumptive zygotes were cultured in the medium supplemented with TIMP-1 ($0.5{\mu}g/ml$), Activin A (100 ng/ml), or HB-EGF (100 ng/ml) at $39^{\circ}C$ in a humidified atmosphere of 5% (v/v) $CO_2$, 5% (v/v) $O_2$ and 90% (v/v) $N_2$. In experiment 2, TIMP-1 + HB-EGF or Activin A + HB-EGF combinations were supplemented in the culture medium. The developmental rate to blastocysts, hatching rate and total cell numbers of the blastocysts were evaluated in both experiments. The embryos cultured in medium without growth factor supplementation was used as control group. In experiment 1, the embryos cultured in medium supplemented with TIMP-1 and Activin A showed significantly higher developmental rate to blastocysts than those cultured with HB-EGF and control (36.9%, 34.1%, 21.2% and 23.1%, respectively) (P<0.0001). However, the hatching rate of blastocyst was significantly higher in embryos with HB-EGF than those with TIMP-1, Actvin A and Control groups (84.4%, 58.8%, 51.4% and 49.3%, respectively) (P<0.001). Total cell number per blastocyst was also significantly higher in embryos with HB-EGF group ($174.3{\pm}2.5$) than those with TIMP-1, Activin A (149.7 and 150.0, respectively) (P<0.05) and Control (119.0) (P<0.001). In experiment 2, embryos cultured with combined treatment of Activin A and HB-EGF resulted in significantly higher rates of blastocysts formation (48.0%), hatching rate (89.7%) and total cell number in blastocyst ($182.3{\pm}2.1$) than those with TIMP-1 and HB-EGF combination group (32.0%, P<0.001; 76.6%, P<0.05; $165.7{\pm}4.2$, P<0.001, respectively). Our data demonstrate that in vitro production of bovine embryos could be improved by combined supplementation of Activin A and HB-EGF in culture medium.

• KSBB Journal
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• v.19 no.5
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• pp.358-362
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• 2004

Characteristics of cell growth and poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] synthesis was investigated through flask and batch cultures of Alcaligenes latus and Comamonas acidovorans. The specific growth rate of C. acidovorans increased with yeast extract concentration and decreased with 1,4-butanediol concentration. Optimum glucose concentration for growth of C. acidovorans was 20 g/L. In one-step flask cultures of C. acidovorans, final dry cell weight and PHA content decreased with the ratio of 1,4-butanediol to glucose, while the 4HB fraction in copolymers gradually increased to 100 $mol\%$ with an initial 1,4-butanediol concentration of 20 g/L as single carbon source. The specific growth rate of A. latus decreased with v-butyrolactone concentration and optimum sucrose concentration for growth was 10 g/L. In batch cultures of A. latus, 4HB fraction increased with initial v-butyrolactone concentration. P(3HB-co-4HB) with 19 $mol\%$ 4HB was obtained when the initial ratio of v-butyloractone (g/L) to sucrose (g/L) was 10 : 10.

• Microbiology and Biotechnology Letters
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• v.17 no.6
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• pp.574-579
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• 1989

Chromosomal DNA fragments of Bacillus sp. YA-14, isolated from soil as a potent $\beta$-xylosidase producing bacterium, were ligated to a vector plasmid pBR322 and used to transfer Escherichia coli HB101 cells. The recombinant plasmid pYXL22 was found to enable the transformants to produce $\beta$-xylosidase. pYXL22 was found to contain the 7.0 kb HindIII DNA fragment originated from the Bacillus sp. YA-14 chromosomal DNA by Southern hybridization. The optimum temperature for the reaction of $\beta$-xylosidase produced by E. coli HB101 (pYXL22) was appeared at 3$0^{\circ}C$. The enzyme was maintained stably up to 4$0^{\circ}C$ when stored 1hr at 4$0^{\circ}C$. The $\beta$-xylosidase was repressed completely by 0.4% (w/v) glucose concentration in E. coli HB101 (pYXL22). The optimum concentration of xylose for the $\beta$-xylosidase production in Bacillus sp. YA-14 was 0.2% (w/v).

• Korean Journal of Exercise Nutrition
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• v.24 no.2
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• pp.30-37
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• 2020

[Purpose] The present study investigated the effect of endurance exercise with blood flow restriction (BFR) performed at either 25% maximal oxygen uptake (${\dot{V}}O_2$ max) or 40% ${\dot{V}}O_2$ max) on muscle oxygenation, energy metabolism, and endocrine responses. [Methods] Ten males were recruited in the present study. The subjects performed three trials: (1) endurance exercise at 40% ${\dot{V}}O_2$ max without BFR (NBFR40), (2) endurance exercise at 25% ${\dot{V}}O_2$ max with BFR (BFR25), and (3) endurance exercise at 40% ${\dot{V}}O_2$ max with BFR (BFR40). The exercises were performed for 15 min during which the pedaling frequency was set at 70 rpm. In BFR25 and BFR40, 2 min of pressure phase (equivalent to 160 mmHg) followed by 1 min of release phase were repeated five times (5 × 3 min) throughout 15 minutes of exercise. During exercise, muscle oxygenation and concentration of respiratory gases were measured. The blood samples were collected before exercise, immediately after 15 min of exercise, and at 15, 30, and 60 minutes after completion of exercise. [Results] Deoxygenated hemoglobin (deoxy-Hb) level during exercise was significantly higher with BFR25 and BFR40 than that with NBFR40. BFR40 showed significantly higher total-hemoglobin (total-Hb) than NBFR40 during 2 min of pressure phase. Moreover, exercise-induced lactate elevation and pH reduction were significantly augmented in BFR40, with concomitant increase in serum cortisol concentration after exercise. Carbohydrate (CHO) oxidation was significantly higher with BFR40 than that with NBFR40 and BFR25, whereas fat oxidation was lower with BFR40. [Conclusion] Deoxy-Hb and total Hb levels were significantly increased during 15 min of pedaling exercise in BFR25 and BFR40, indicating augmented local hypoxia and blood volume (blood perfusion) in the muscle. Moreover, low-and moderate-intensity exercise with BFR facilitated CHO oxidation.

• Asian-Australasian Journal of Animal Sciences
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• v.18 no.11
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• pp.1642-1645
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• 2005

This study was conducted to investigate the effects of Biotite V supplementation on growth performance, nutrients digestibility and blood constituents and to evaluate whether Biotite V could replace an antibiotics in growing pigs diet. One hundred twenty pigs with initial body weight of 18.35${\pm}$0.15 kg were used in a 28 days growth trial. Pigs were allotted to four treatments by sex and body weight in a randomized complete block design. There were six replicate pens per treatment and five pigs per pen. Four dietary treatments were: 1) NC (basal diet without antibiotics), 2) PC (basal diet+0.1% CTC), 3) NCBV (NC diet+0.5% 200 mesh Biotite V) and 4) PCBV (PC diet+0.5% 200 mesh Biotite V). Through the entire experimental period, ADG tented to increase in NCBV and PCBV treatments compared to NC and PC treatments respectively, but no significant differences were observed (p>0.05). ADFI was slightly lower in NCBV and PCBV treatments than that in NC and PC treatments without significant differences (p>0.05). Gain/feed in PC and PCBV treatments was improved significantly compared to NC treatment (p<0.05). N and Ca digestibilities were higher in PCBV treatments than those in PC treatment (p<0.05). DM and P digestibilities were not affected by the addition of Biotite V (p>0.05). RBC, HCT, Hb, lymphocyte and monocyte were increased numerically in NCBV and PCBV treatments compared to NC and PC treatments (p>0.05). WBC was lower in treatment groups than that in NC treatment, but no significant differences were observed (p>0.05). In conclusion, dietary supplementation of Biotite V can better the gain/feed and some of the nutrients digestibilities in growing pigs. It has a possibility to replace antibiotics in swine diet.

• Bulletin of the Korean Chemical Society
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• v.30 no.3
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• pp.582-588
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• 2009

Direct electron transfer of hemoglobin (Hb) in the chitosan (CTS) and $TiO_2$ nanoparticles (nano-$TiO_2$) composite films was achieved by using a room temperature ionic liquid of 1-butyl-3-methylimidazolium hexafluorophosphate ($BMIMPF_6$) modified carbon paste electrode (CILE) as the basal electrode. UV-Vis and FT-IR spectroscopy indicated that Hb in the film retained the native structure. Electrochemical investigation indicated that a pair of well-defined quasi-reversible redox peaks of Hb heme Fe(III)/Fe(II) was obtained with the formal potential located at -0.340 V (νs. SCE) in pH 7.0 phosphate buffer solution (PBS). The electrochemical parameters such as the electron transfer coefficient (α), the electron transfer number (n) and the standard electron transfer rate constant ($k_s$) were got as 0.422, 0.93 and 0.117 $s^{-1}$, respectively. The fabricated CTS/nano-$TiO_2$/Hb/CILE showed good electrocatalytic ability to the reduction of trichloroacetic acid (TCA) and hydrogen peroxide ($H_2O_2$), which exhibited a potential application in fabricating a new kind of third generation biosensor.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.11
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• pp.1462-1468
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• 2010

The effects of vitamin C and vitamin E with selenium on acid-base balance and some stress hormones were evaluated during heat stress in goats. Goats, 1.5 years of age, were divided into control, heat stress and antioxidant treatment groups 1, 2 and 3. Except for the control, all groups were exposed to a temperature of $40{\pm}2^{\circ}C$ with a relative humidity of 30% for 5 h/d for 21 days in a psychrometric chamber. Rectal temperature and respiratory rates were recorded daily post exposure. Blood samples were collected on every 3rd day for estimation of plasma vitamins C and E, total antioxidant activity and hormones, and separate blood samples were taken to estimate acid-base status. The rectal temperature and respiratory rates were increased (p<0.05) in the heat stress group only. Except for pH and $pO_2$, which were increased significantly (p<0.05) other parameters of acid-base balance such as $pCO_2$, $HCO_3^-$, $TCO_2$, BEb, BEcef, PCV and Hb were significantly decreased (p<0.05) in the heat stress group. An improvement in acid-base status was noted in the antioxidant supplemented groups. Prolactin and cortisol levels were significantly (p<0.05) higher and free T3 and T4 levels were significantly (p<0.05) lower in the heat stress group. Levels of prolactin and cortisol were decreased and free T3 and T4 were increased in antioxidant treatment groups. Different levels of antioxidant supplementation resulted in similar protection against heat stress.

• The Korean Journal of Nuclear Medicine
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• v.38 no.1
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• pp.99-108
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• 2004

Purpose: The ability to noninvasively track the migration of neural progenitor cells would have significant clinical and research implications. We generated stably transfected F3 human neural progenitor cells with human sodium/iodide symporter (hNIS) for noninvasively tracking F3. In this study, the expression patterns of hNIS gene in F3-NIS were examined according to the cultured time and the epigenetic modulation. Materials and Methods: F3 human neural stem cells had been obtained from Dr. Seung U. Kim (Ajou University, Suwon, Korea). hNIS and hygromycin resistance gene were linked with IRES (Internal Ribosome Entry Site) under control of CMV promoter. This construct was transfected to F3 with Liposome. To investigate the restoration of hNIS gene expression in F3-NIS, cells were treated with demethylating agent (5-Azacytidine) and Histone deacetylase inhibitor (Trichostatin A: TSA). The expression of hNIS was measured by I-125 uptake assay and RT-PCR analysis. Results: The iodide uptake of the F3-NIS was higher 12.86 times than F3 cell line. According to the cell passage number, hNIS expression in F3-NIS gradually diminished. After treatment of 5-Azacytidine and TSA with serial doses (up to $20{\mu}M$, up to 62.5nM, respectively) for 24 hours, I-125 uptake and mRNA of hNIS in F3-NIS were increased. Conclusion: These results suggest that hNIS transfected F3 might undergo a change in its biological characters by cell passage. Therefore, the gene ex[ressopm of exogenous gene transferred human stem cell might be affected to the epigenetic modulation such as promoter methylation and Histone deacetylation and to the cell culture conditions.