• Clinical and Experimental Reproductive Medicine
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• v.21 no.3
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• pp.241-245
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• 1994

Despite increasing success rate of IVF, poor response to ovarian stimulation remains a problem. So, attempts to improve ovarian responses, for example, by using combined gonadotropin-releasing hormone analogue(GnRH-a) and human menopausal gonadotropin(hMG) have shown limited success. It is reported that response of granulosa cells in vitro to FSH is stimulated by co-incubation with IGF-l, and IGF-l production can be increased by growth hormone. This suggest that combination regimen of G.H. and hMG may augment follicle recruitment. In fifteen patients who had previous history of poor ovarian response to gonadotropin stimulation after pituitary suppression with mid -luteal GnRH-a, the effectiveness of cotreatment with G.H. in IVF program was evaluated using a combination regimen of G.R. and hMG at Korea University Hospital IVF Clinic. Ovarian responses to gonadotropin stimulation in control and GH-treated cycles assessed by total dose and duration of hMG treatment, follicular development and peak $E_2$ level, number of eggs retrieved, and fertilization rates were also assessed. In each group, serum and follicular fluid IGF-1 concentrations on day of egg collection were measured by RIA after acidification and extraction by reveresed phase chromatography. Patients receiving G.H. required fewer days and ampules of gonadotropins, developed more oocytes, and more embryos transferred. But, the differences were not statistically significant, except the duration of hMG treatment. Our data showed a significantly higher concentration of IGF-l in the serum, not in the follicular fluid, of patients treated with G.H. compared with control group. These data suggest that growth hormone treatment does not improve the ovarian response in women with limited ovarian reserve to gonadotropin stimulation for IVF.

• Journal of Microbiology and Biotechnology
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• v.29 no.5
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• pp.765-775
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• 2019

A new ${\alpha}$-amylase-encoding gene (amySL3) of glycoside hydrolase (GH) family 13 was identified in soda lake isolate Alkalibacterium sp. SL3. The deduced AmySL3 shares high identities (82-98%) with putative ${\alpha}$-amylases from the genus Alkalibacterium, but has low identities (<53%) with functionally characterized counterparts. amySL3 was successfully expressed in Escherichia coli, and the recombinant enzyme (rAmySL3) was purified to electrophoretic homogeneity. The optimal temperature and pH of the activity of the purified rAmySL3 were determined to be $45^{\circ}C$ and pH 7.5, respectively. rAmySL3 was found to be extremely halophilic, showing maximal enzyme activity at a nearly saturated concentration of NaCl. Its thermostability was greatly enhanced in the presence of 4 M NaCl, and it was highly stable in 5 M NaCl. Moreover, the enzyme did not require calcium ions for activity, and was strongly resistant to a range of surfactants and hydrophobic organic solvents. The major hydrolysis products of rAmySL3 from soluble starch were maltobiose and maltotriose. The high ratio of acidic amino acids and highly negative electrostatic potential surface might account for the halophilic nature of AmySL3. The extremely halophilic, calcium-independent, and surfactant-resistant properties make AmySL3 a promising candidate enzyme for both basic research and industrial applications.

• Journal of Korean Medicine Rehabilitation
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• v.18 no.2
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• pp.97-109
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• 2008

Objectives : Effects of Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) on intellectual development and learning ability were investigated growth and intellectual impairment rat with insufficient nutrition diet. Methods : We divided male Sprague-Dawley rats into 4 groups(A, B, C, D). They were normal group(A), growth deficiency rat with insufficient nutrition diet group(B), growth deficiency rat with 0.1% Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) group(C) and growth deficiency rat with 0.2% Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) group(D). They were administered for 5 weeks. We measured body weight, serum growth hormone, insulin-like growth factor and thyroid stimulating hormone, RBC, concentration of Hb and PCV ratio, total WBC and its composition, the values of plasma glutamic oxaloacetic transaminase(GOT) and glutamic pyruvic transaminase(GPT) activities and morris water maze test in escape distance, escape time and escape speed. Results : 1. Body weight showed a tendency to increase in the Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) groups and 0.2% Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) group showed significantly different than control groups. 2. Serum growth hormone, insulin-like growth factor and thyroid stimulating hormone showed a tendency to increase in Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) groups, however these values showed no significantly different. 3. About the counts of RBC, 0.1% and 0.2% Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) groups showed significantly different than control groups. Concentration of Hb was higher than control group in Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) groups. And 0.2% Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) group showed significantly different than control groups in PCV ratio. 4. The counts of total WBC and its composition showed no significantly different in all treatment groups. 5. The values of plasma glutamic oxaloacetic transaminase(GOT) and glutamic pyruvic transaminase(GPT) activities showed no significantly different in all treatment groups. 6. In the morris water maze test, in escape distance and escape speed, Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) groups showed no significantly different than control group. But Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) groups showed the increasing tendency. Conclusions : So Kyungohkgo($Qi{\acute{o}}ng\;y{\grave{u}}\;g{\grave{a}}o$) have an effect of promoting growth of rats and might be effect to treat various kinds of growth delay in children.

• The Korean Journal of Pesticide Science
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• v.20 no.4
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• pp.326-340
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• 2016

GHS was compared with that of current RDA directives in skin and eye irritation for plant protection products (PPPs). According to the result of evaluation and classification of skin and eye irritation by RDA directives and GHS for 59 PPPs, 25.5% of PPPs were changed the classification of skin irritation from slightly and moderately by RDA directives to Not Classified by GHS, and 45.8% of PPPs were changed the classification of eye irritation from slightly by RDA directives to Not Classified by GHS. 6.8% of PPPs in skin irritation and 5.1% in eye irritation were classified more highly as category 1 by GHS because of irreversible effect. According to the result of comparison with GHS classification by data of irritation study for PPPs and by GHS classification and concentration of active substances, 25.5% was changed more lowly and 3.4% more highly in skin irritation, and 25.4% was changed more lowly and 11.9% more highly in eye irritation. It is needed to draw a plan to supplement for other results between GHS and RDA directives.

• Korean Journal of Animal Reproduction
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• v.26 no.2
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• pp.153-163
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• 2002

In vitro matured porcine oocytes have very small volume of perivitellinspace (PVS). In these respect, the effect of sucrose and polybrene on the efficiency of gene transfer was investigated. As a gene (hGH) transfer vehicle, vesicular stomatitis virus glycoprotein pseudotyped retroviral vector (VSV-G) was used. Sucrose treatment has no detrimental effect on the rates of cleavage and resulted in the enlargement of PVS for the efficient introduction of retroviral vector stocks. Introduction rates of retrovirus in 0.5, 1, 2, 3 % sucrose treatment group were higher than that of the non-treatment group (39.3, 43.3, 35.7, 40.7 % vs. 8.3 %), respectively. In addition, we observed that sucrose pretreatment during injection procedure significantly reduce the frequency of polyspermy. In general, polybrene is a polycation essential for retrovirus transduction. The groups with the addition of 0.5, 5, 50$\mu\textrm{g}$/$m\ell$ polybrene exhibited a significant effect on gene transfer compared to that of the non-addition group (56.5, 50.0, 57.1 % vs. 34.6 %), respectively But, when the oocytes were co-injected with retrovirus and 50$\mu\textrm{g}$/$m\ell$ polybrene, the rates of cleavage and blastocyst development were 43.3 and 4.6%, respectively. This rates were lower than those of the non-addition group (70.0 and 17.3 %). In conclusion, sucrose pretreatment have increased efficiency of retroviral mediated gene transfer in porcine oocytes with no damage on in vitro fertilization and embryo development. In addition, sucrose pretreatment was beneficial in polyspermy inhibition. Presence of polybrene during microinjection showed a beneficial effect on the gene transfer in porcine oocytes, in low concentration. And these results will provide an useful tool for production of transgenic pigs by retroviral mediated gene transfer.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.10
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• pp.1411-1415
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• 2008

The purpose of the present study was to determine the onset of breeding season, the occurrence of silent and true heats and the duration of estrus in female Markhoz goats of the Kurdistan province in Iran. Ten, 3 years-old Markhoz does with an average weight of $34.05{\pm}2.62kg$ and with one kidding record, were used. The goats were maintained in an open barn under constant nutritional levels and natural photoperiod. One aproned buck was used twice a day every 12 h to detect estrus from mid August to early January. For the determination of the onset of reproductive activity as well as occurrence of silent heat, blood samples were collected every 10 days, from the beginning of the experiment. After $2^{nd}$ standing heat, blood samples were obtained twice a week in order to assess luteal activity and the length of estrous cycles. In this study, estrous behavior was observed including sniffing, vocal exchange, following courtship, flehman, standing heat and mounting. The results of the progesterone assay indicated that in this goat silent heats occur mostly in the early breeding season. The first standing heat was observed in mid-October which was considered as the onset of the breeding season. Duration of the estrous cycle and estrus was recorded as being $20.93{\pm}1.56days$ and $38.86{\pm}15.19h$, respectively. The correlation coefficient between length of daylight and occurrence of estrus was negative (r = -0.470) but not significant (p>0.05). The data showed that there was no significant effect of body weight on estrous cycles (first, second and third) and estrous periods (first, second, third and fourth). Progesterone levels were not significantly different in the first, second and third estrous cycles at days 0, 4, 10, 12, 14 and 19. The results of progesterone assay during the estrous cycle indicate that follicular and luteal phases last 4-5 and 14-15 days, respectively and the concentration of serum progesterone in these phases was $0.88{\pm}0.08$ and $7.44{\pm}0.26ng/ml$, respectively. The study concluded that Markhoz does could be considered as a breed with a short breeding season and an optimal estrous activity in autumn.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.11
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• pp.1504-1512
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• 2009

A feeding trial was conducted to examine the effect of high-temperature-micro-time (HTMT) processing of diets containing extruded soybean (ESB) in high quantity on milk fat production, metabolic responses, and the formation of conjugated linoleic acid (CLA) and trans-vaccenic acid (TVA). Twenty-one multiparous Holstein cows in mid-lactation were blocked according to milk yield in the previous lactation. Cows within each block were randomly assigned to either normal concentrate or HTMT treated diets containing ESB (7.5% HTMT-ESB and 15% HTMT-ESB). It was hypothesized that the HTMT-ESB would affect the undegradable fatty acids in the rumen and, thus, would modify the fatty acid profile of milk fat. Both 7.5% and 15% HTMT-ESB did not affect milk yield, fat, protein, lactose and solid-not-fat (SNF), but the proportion of cis-9, trans-11 CLA in milk fat was significantly increased by these treatments. Content of TVA in milk fat was not affected by HTMT-ESB. The HTMT-ESB influenced the fatty acid profile in milk fat, but there was little difference between 7.5% and 15% of supplementation. HTMT-ESB feeding significantly decreased the concentration of plasma insulin and glucose, while plasma growth hormone (GH), triglyceride (TG), non-esterified fatty acid (NEFA) and HDLcholesterol were increased by 7.5% and 15% ESB-HTMT supplementation in comparison to the control group (p<0.05). However, no significant difference was observed in plasma LDL-cholesterol, insulin like growth factor (IGF)-1, T3, T4, and leptin concentrations among treatments (p>0.05). The present results showed that cis-9, trans-11 CLA production was increased by HTMT treatment of dietary ESB without reduction of milk fat, and the unchanged milk fat and yield was assumed to be associated with the constant level of thyroid hormones, leptin, and IGF-1.

• Journal of Microbiology and Biotechnology
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• v.23 no.11
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• pp.1519-1528
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• 2013

Bacillus pumilus SG2, a halotolerant strain, expresses two major chitinases designated ChiS and ChiL that were induced by chitin and secreted into the supernatant. The present work aimed to obtain a mutant with higher chitinolytic activity through mutagenesis of Bacillus pumilus SG2 using a combination of UV irradiation and nitrous acid treatment. Following mutagenesis and screening on chitin agar and subsequent formation of halos, the mutated strains were examined for degradation of chitin under different conditions. A mutant designated AV2-9 was selected owing to its higher chitinase activity. To search for possible mutations in the whole operon including ChiS and ChiL, the entire chitinase operon, including the intergenic region, promoter, and two areas corresponding to the ChiS and ChiL ORF, was suquenced. Nucleotide sequence analysis of the complete chitinase operon from the SG2 and AV2-9 strains showed the presence of a mutation in the catalytic domain (GH18) of chitinase (ChiL). The results demonstrated that a single base change had occurred in the ChiL sequence in AV2-9. The wild-type chitinase, ChiL, and the mutant (designated ChiLm) were cloned, expressed, and purified in E. coli. Both enzymes showed similar profiles of activity at different ranges of pH, NaCl concentration, and temperature, but the mutant enzyme showed approximately 30% higher catalytic activity under all the conditions tested. The results obtained in this study showed that the thermal stability of chitinase increased in the mutant strain. Bioinformatics analysis was performed to predict changes in the stability of proteins caused by mutation.

• Korean Journal of Plant Tissue Culture
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• v.21 no.1
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• pp.29-34
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• 1994

The possibility that DNA could move out of the single cells isolated from geranium (Pelargonium zonale hybrids, 'PintoScarlet') callus was determined by the elechophoretic DNA analysis after treatment of low pH, various concentrations of KNO$_3$, 2,4-D, and GA$_3$ followed by the centrifugal force, all of which are hewn to and the physico-chemical properties of the cell wall. The centrifugal force of l,800 xg was need for DNA migration after the above treatment, but 7k300 xg was required without the treatments. In this experiment the optimum concentration (300 mg/L) of sodium dodecyl sulfate (SDS) used as an anion detergent to collect the negatively charged DNA was very critical not to damage the cell wall It can be concluded that the centrifugal force played a key role for the DNA migration through the cell wall, and the treatments of low pH (4.0), 0.5% KNO$_3$, 1.5 mg/L GA$_3$and 1mg/L 2,4-D further increased the DNA migration.

• Applied Biological Chemistry
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• v.39 no.5
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• pp.333-337
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• 1996

For continuous production of galactooligosaccharides(GOS), $\beta-galactosidase$ with h1gh transgalactosylation activity from Bacillus sp. A 4442 was Immobilized onto $Diaion^{TM}$ HPA 75(styrene-divinylbenzene resin). The parameters influencing enzyme immobilization were scrutinized in order to maximize immobilization yield while minimizing enzyme inactivation. The optimum conditions turned out to be: Tris buffer concentration 30 mM, pH 8.0, contact time at room temperature 3 hr, and enzyme loading 25 mg protein/g resin. Both the thermal stability and the operational stability of immobilized enzyme were markedly enchanced by the treatment with 0.5% glutaraldehyde as a cross-linker. Under the experimental conditions established, the yield of ${\beta}-galactosidase$ immobilization was 40% or more and the activity of the immobilized enzyme ca. 200 U/g resin. When a packed-bed reactor was employed to continuously convert lactose to GOS, the specific production, which refers to as the amount of commercially valuable GOS produced by a unit amount of immobilized ${\beta}-galactosidase$, was found to be ca. 300 g GOS/g carrier.