• Toxicological Research
• /
• v.33 no.2
• /
• pp.107-118
• /
• 2017

Although alternative test methods based on the 3Rs (Replacement, Reduction, Refinement) are being developed to replace animal testing in reproductive and developmental toxicology, they are still in an early stage. Consequently, we aimed to develop alternative test methods in male animals using mouse spermatogonial stem cells (mSSCs). Here, we modified the OECD TG 489 and optimized the in vitro comet assay in our previous study. This study aimed to verify the validity of in vitro tests involving mSSCs by comparing their results with those of in vivo tests using C57BL/6 mice by gavage. We selected hydroxyurea (HU), which is known to chemically induce male reproductive toxicity. The 50% inhibitory concentration ($IC_{50}$) value of HU was 0.9 mM, as determined by the MTT assay. In the in vitro comet assay, % tail DNA and Olive tail moment (OTM) after HU administration increased significantly, compared to the control. Annexin V, PI staining and TUNEL assays showed that HU caused apoptosis in mSSCs. In order to compare in vitro tests with in vivo tests, the same substances were administered to male C57BL/6 mice. Reproductive toxicity was observed at 25, 50, 100, and 200 mg/kg/day as measured by clinical measures of reduction in sperm motility and testicular weight. The comet assay, DCFH-DA assay, H&E staining, and TUNEL assay were also performed. The results of the test with C57BL/6 mice were similar to those with mSSCs for HU treatment. Finally, linear regression analysis showed a strong positive correlation between results of in vitro tests and those of in vivo. In conclusion, the present study is the first to demonstrate the effect of HU-induced DNA damage, ROS formation, and apoptosis in mSSCs. Further, the results of the current study suggest that mSSCs could be a useful model to predict male reproductive toxicity.

• The Korean Journal of Zoology
• /
• v.39 no.1
• /
• pp.36-46
• /
• 1996

An endogenous phenoloxidase (EPO) from earthworm, Lumbricus rubellus, has been purified and characterized. The purified EPO using ammonium sulfate fractionation, Blue-2, Phenyl-, and Q-sepharose chromatography steps was revealed in SDS-PAGE as a single protein banri with Mr. of 59 kl)a. A native strudure of the enzyme was examined with an in situ staining of a nondenatudng-PAGE using DL-dopa as a substrate. The result showed that a single band due to the EPO activity was located siighdy above a standard polypeptide with Mr. of 210 kl)a. These fads indicate that the EPO is an oligomeric enzyme. The presence of a monophenolase activity of the purified EPO, which hydroxylates tyrosine to dopa, was confirmed by observing dopachrome accumulation at 475 nm at PH 8.0 with a typical lag phase during 60 mm. of meausrement. A series of inhibition study has been performed for the enzyme with several divalent cation chelators such as phenyithiourea (Flu), 1, lO-phenanthroline, EDTA, and EGTA. Among them, only V'flj inhibited the enzyme with 1C0.5 of 65 MM, which indicated that copper was critical for the catalysis of EPO. The enzyme was maximally active at 35'C and pH 8.0 when L-dopa to dopachrome conversion was spectrophotometricaily monitored at 475 nm. The apparent Km values of P0 for L-opa were obtained as 1.86 mM and 13.8 mM at pH 6.5 and 8.0, respectively. The catalytic efficiencies at both pH were almost identical [(kat/Km)pH8.0/(kcat/Km)pH6.5 = O.92] while the Vmax at p11 8.0 was 6.6-fold higher than that at pH 6.5. This fact may indicate that pH affeds the catalysis at substrate and/or enzyme-substrate complex level rather than the enzyme itself. Taken together, the EPO was an oligomeric enzyme which did not require proteolysis for its activation. These results also indicated that the enzyme can exist, at least, in part as a latent form In vivo, which might be distinct from the prophenoloxidase activating system. Therefore, it is pertinent to consider that there must be certain regulatory molecules or phenomena in L. rubellus which make the 1,0 in a latent form in vivo before the foreign invasions.

• Journal of Ginseng Research
• /
• v.30 no.2
• /
• pp.88-94
• /
• 2006

Ginseng as a raw material for production of probiotic ginseng product by lactic acid bacteria (LAB) was evaluated in this study. Either white ginseng (WG) or red ginseng (RG) (1% or 5%, w/v) were directly inoculated with a 24 hold seed culture of twenty seven substrains of four different LAB ($1.0{\times}10^6CFU/ml$); Lactobacillus spp., Streptococcus/Enterococcus spp., Leuconostoc/Lactococcus spp. and Bifidobacterium spp., and incubated at $37^{\circ}C$ for 24 or 48 h. Among 27 kinds of LAB, seven substrains of Lactobacillus (MG208, MG311, MG315, MG501, MG501C, MG505, MG590) and one Bifidobacterium (MG723) were selected based on their dose dependent stimulation of the growth of LAB in the presence of ginseng and changes in pH, acidity and viable cell counts during fermentation were examined. Lactobacillus MG208 specifically was found to show the best growth on 5% RG and reached nearly $14.0{\times}10^8CFU/ml$ after 48 h of fermentation and produced the titratable acidity as $0.84{\pm}0.02%$, whereas the pH was significantly lowered from $6.80{\pm}0.01\;to\;3.42{\pm}0.02$. These results indicated that ginseng can be an appropriate material to prepare the fermentation product by several strains of LAB. Therefore we should further check whether probiotic ginseng product may have synergistic health benefits of both probiotics and ginseng to serve for vegetarians and lactose-allergic consumers.

• The Korean Journal of Physiology
• /
• v.21 no.2
• /
• pp.241-257
• /
• 1987

The effects of external $Ca^{2+}$ and $Ca^{2+}-antagonists$ on the spontaneous contractions and electrical activities were investigated in guinea-pig stomach in order to clarify the mechanism for the generation of slow waves. Electrical responses of circular smooth muscle cells were recorded using glass capillary microelectrodes filled with 3 M KCl. All experiments were performed in tris-buffered Tyrode solution which was aerated with 100% $O_2$ and kept at $35^{\circ}C$. The results obtained were as follows: 1) The amplitude of spontaneous contractions was maximal at around 2-4 mM $Ca^{2+}$, whereas their frequency was inversely related with external $Ca^{2+}$ within the range of 0.5 to 16 mM $Ca^{2+}$. 2) Verapamil suppressed the amplitude of spontaneous contraction in a dose-dependent manner, while the frequency of spontaneous contractions was almost not changed over the whole concentration of verapamil $(0.01{\sim}5\;mg/l)$. 3) Manganese increased both the amplitude and the frequency of spontaneous contractions dose-dependently in low $Mn^{2+}$ (below 0.05 mM $Mn^{2+}$), while their amplitude and frequency were decreased in high $Mn^{2+}$ (above 0.1 mM $Mn^{2+}$). 4) The ampltude and maximum rate of rise of slow waves were incrased in high $Ca^{2+}$ solution. In $Ca^{2+}-free$ solution, the spontaneous contractions recorded simultaneously with slow waves ceased and tonic contraction ($Ca^{2+}-free$ contracture) was developed in parallel with membrane depolarization and the disappearance of slow waves. 5) Verapamil (1 mg/1) decreased the amplitude and maximum rate of rise of slow waves and it depolarized the membrane by about 6 mV, whereas the frequency of slow waves was not affected by verapamil. 6) Manganese showed different characteristic effects between low and high $Mn^{2+}$ on the slow waves: In low $Mn^{2+}$ (0.05 mM $Mn^{2+}$), the initial rapid increases and the subsequent gradual decreases in three parameters of slow waves (amplitude, rate of rise, and frequency of slow waves) till a new steady state were observed. However, in high $Mn^{2+}$ (0.5 mM $Mn^{2+}$) slow waves disappeared and membrane was depolarized. From the above results, the following conclusions could be made: 1) $Ca^{2+}$ is necessary for a generation of the slow waves, even though it is small amount. 2) Verapamil suppresses the spontaneous contractions of gastric antral strip by the decreases in amplitude and maximum rate of rise of slow waves, while this drug does not block the $Ca^{2+}-channel$ involved in the generation of slow waves. 3) Manganese has dual actions on the $Ca^{2+}-channels$; the $Ca^{2+}-channel$ involved in the generation of slow waves (or Na-Ca exchange system) or the channel for the generation of spike potentials are stimulated by a low concentration of $Mn^{2+}$, while both the $Ca^{2+}$. Channels are blocked by high concentration of $Mn^{2+}$.

• Fire Science and Engineering
• /
• v.27 no.2
• /
• pp.25-30
• /
• 2013

'Heaundae Woosin Building' fire was the case that a fire breaking out on the $4^{th}$ floor spread out the $38^{th}$ floor which is a top story and the penthouse was destroyed by fire. After this fire, the fire safety for high rise buildings has been on the rise and several new laws and regulations related in the buildings were created. The study is to analyze Heaundae officetel building case using FDS which is one of the CFD programs for fire. The methodology of this study is to analyze the case comparing with fire spread and route from a virtual fire simulation and related articles and a video clip of actual scene fire. This study shows that a fire spreading on top of levels spent approximately 30 minutes and, which is similar to the actual fire case. Also the pattern of spread has similarity with the case. However, even if the actual fire case shows the fire pattern was "V shape", the smoke-view presents the fire dose not spread horizontally as much as the real fire case. The result shows uncertainty of the modeling based on many grids and a limitation of putting interior finish input sources and the direction of the wind might cause the difference. Also, to analyze factors influencing on a vertical fire, another fire modeling is performed by in condition of modeling environment considering concrete interior finish between buildings and no wind. The result presents the fire spread in smoke-view does not spread vertically as much as the actual case.

• Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
• /
• 2009.11a
• /
• pp.114-114
• /
• 2009

ZnO has received considerable attention due to its potential applicability to optoelectronic devices such as ultraviolet-light emitting diodes (UVLEDs) and laser diodes (LDs). As well known, however, polar ZnO with the growth direction along the c-axis has spontaneous and piezoelectric polarizations that will result in decreased quantum efficiency. Recently, nonpolar ZnO has been studied to avoid such a polarization effect. In order to realize applications of nonpoar ZnO-based films to LEDs, growth of high quality alloys for quantum well structures is one of the important tasks that should be solved. $Mg_xZn_{1-x}O$ and $Cd_xZn_{1-x}O$ is ones of most promising alloys for this application because the alloys of ZnO with MgO and CdO provide a wide range of band-gap engineering spanning from 2.4 to 7.8 eV. In this study, we investigated on $Mg_xZn_{1-x}O$ films grown with various Mg/Zn flux ratios The films were grown on R-plane sapphire substrates by plasma-assisted molecular beam epitaxy (PAMBE). we investigated on $Mg_xZn_{1-x}O$ films grown with various Mg/Zn flux ratios. The films were grown on R-plane sapphire substrates by plasma-assisted molecular beam epitaxy (PAMBE). With the relatively low Mg/Zn flux ratios, a typical striated anisotropic surface morphology which was generally observed from the nonpolar (11-20) ZnO film on r-plane sapphire substrates. By increasing the Mg/Zn flux ratio, however, additional islands were appeared on the surface and finally the surface morphology was entirely changed, which was generally observed for the (0001) polar ZnO films by losing the striated morphology. Investigations by X-ray $\Theta-2{\Theta}$ diffraction revealed that (0002) and (10-11) ZnO planes are appeared in $Mg_xZn_{1-x}O$ films by increasing the Mg/Zn flux ratio. Further detailed investigation by transmission electron microscopy (TEM) and photoluminescence (PL) will be discussed.

• Asian-Australasian Journal of Animal Sciences
• /
• v.16 no.10
• /
• pp.1443-1450
• /
• 2003

Crossbred (Bos taurus${\times}$Bos indicus) calves were used from birth till 14 weeks of age to evaluate three sources of protein that differed in ruminal degradability viz. groundnut cake alone (HD) or in combination with cottonseed meal (MD) and meat and bone meal (LD), when fed along with two sources of non-structural carbohydrates viz. raw (R) and thermally processed (P) maize. Twenty four new born calves were arranged in six groups in a $3{\times}2$ factorial design and fed on whole milk up to 56 d of age. All the different calves received calf startes along with green oats (Avena sativa) from 14 d of age onwards free-choice. A metabolism trial of 6d starters duration, conducted after 90 d of experimental feeding, revealed greater (p<0.05) digestibility of DM, OM, total carbohydrates, NDF and ADF in calves fed on the P diets than on the R diets promoting greater (p<0.05) metabolizable energy intake. The digestibility of NDF was higher (p<0.01) on LD diets where as calves on MD diets exhibited significantly lower digestibility of ADF (p<0.01). The retention of nitrogen per unit metabolic body size was significantly (p<0.05) higher on the LD-P diet than on the diet HD-P which, in turn, was higher (p<0.05) than that of HD-R. Nitrogen retention as percentage of intake was significantly greater (p<0.05) on LD-P than on LD-R diets (52.2 vs. 36.4%). Also, P fed calves utilized nitrogen more efficiently than the R fed as shown by retention of significantly greater proportions of intake (47.4 vs. 40.9%) and absorbed (65.8 vs. 59.5%) nitrogen. Calorimetric evaluation of the diets through open-circuit respiration chamber revealed that the dietary treatments had no impact on methane production by calves. The intake of DE and ME was improved (p<0.01) because of maize processing resulting in greater (p<0.01) retention of energy. The protein degradability exerted no influence on the partitioning or retention of energy. A significant interaction between cereal and protein types was evident with respect to retention of both nitrogen (p<0.01) and energy (p<0.05). In conclusion, no discernible trend in the influence of cereal processing was apparent on the dietary protein degradability, but the positive effect of cereal processing on energy retention diminished with the increase in dietary undegradability.

• Journal of the East Asian Society of Dietary Life
• /
• v.23 no.1
• /
• pp.53-60
• /
• 2013

The preventive actions of methanol extracts from Ribes diacanthum Pall. (RDP) against high blood glucose levels in alloxan-induced diabetic mice were investigated. ICR mice (male, 8 wks) were divided into four experimental groups: a normal group (N), an alloxan-induced group (control), a RDP 1 mg/kg b.w./day + alloxan group (RDP-1) and a RDP 3 mg/kg b.w./day + alloxan group (RDP-2). Mice were fed RDP extracts for 14 days and then a diabetic condition induced by injecting alloxan (50 mg/kg b.w. i.v.). The total phenolic contents of RDP were 0.508 mg/g. Plasma glucose levels were significantly lower in RDP groups (RDP+alloxan) than the control (alloxan-induced diabetic group). In an oral glucose tolerance test, the glucose levels of RDP extract groups were significantly decreased compared to the control group. The levels of total cholesterol, triglycerides, AST, ALT, uric acid, and blood urea nitrogen in serum were lower in the RDP groups than the control group, whereas levels of HDL-cholesterol showed no difference between treatment groups. We conclude that RDP extracts positively influence blood glucose levels in diabetic mice.

• Korean Journal of Plant Resources
• /
• v.29 no.4
• /
• pp.419-433
• /
• 2016

A floristic study was conducted 23 times from March 2010 to May 2016 on Mt. Yongbongsan and Mt. Suamsan region of Chungcheongnam-do which lies across Sapgyo-eup, Deoksan-myeon Yesan-gun, and Hongbuk-myeon Hongseong-gun. Based on voucher speciemens, list of vascular plants found in this area were contained 532 taxa in total including 105 families, 312 genera, 468 species, 4 subspecies, 47 varieties, and 13 forma. The 8 taxa were Korean endemic plants such as Populus tomentiglandulosa T.B.Lee, Clematis brachyura Maxim, C. trichotoma Nakai, Indigofera koreana Ohwi, Forsythia koreana (Rehder) Nakai, Paulownia coreana Uyeki, Weigela subsessilis (Nakai) L.H.Bailey, Aster koraiensis Nakai. 4 taxa of rare plants designated by the Korea Forest Service such as Taxus cuspidata Siebold & Zucc., Lithospermum arvense L., Allium senescens L., and Iris ensata var. spontanea (Makino) Nakai. Endangered plants designated by the Ministry of Environment was not found in this study. The 11 taxa of floristic regional indicator plants in 3^rd to 5^th degree were found. 68 taxa naturalized plants were found and the naturalized ratio was 12.8% while 21.2% of urbanized ratio which is regarded quite high.

• Journal of the Korean Chemical Society
• /
• v.47 no.4
• /
• pp.391-400
• /
• 2003

This study was performed the analysis of seven kinds of the hight school chemistry II textbooks based on the 6th curriculum. Particularly, inquiry activity part was analyzed by the three dimension framework which consists of inquiry content dimension, inquiry process dimension and inquiry context dimension. In the analysis of the inquiry content dimension of inquiry activities, the total number of themes in seven kinds of textbook was 212. And the number of inquiry activities in seven kinds of textbook was diverse: A textbook had 28, B textbook 25, C textbook 31, D textbook 35, E textbook 31, F textbook 29 and G textbook 33. As for the avaerage number of inquiry activities of each chapter, chapter I "Material Science" is 3.00(9.91${\%}$), chapter II "Atomic Structure and Periodic Table" 4.57(15.1${\%}$), chapter III "Chemical Bonding and Compound" 6.86(22.6${\%}$), chapter IV "State of Matter and Solution" 7.00(23.1${\%}$), chapter V "Chemical Reaction" 8.86(29.2${\%}$). For the analysis of inquiry process dimension, it follows in the order of 'observation and measuring (66.7${\%}$)', 'Interpreting data and formulating generalizations (26.5${\%}$)', 'seeing a problem and seeking ways to solve it (4.1%)', and 'building, testing and revising the theoretical model (2.7${\%}$)'. As for the analysis of the inquiry context dimension, the scientific context occupied 90.5${\%}$, the individual context 4.3${\%}$, the social context 0.9${\%}$, and the technical context 4.3${\%}$. It shows that the proportion of STS(Science-Technology-Society) related contents in inquiry activities was only 9.5${\%}$.