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

• Journal of Bio-Environment Control
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• v.11 no.4
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• pp.181-187
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• 2002

The overall objective of this study was to improve tomato fruit quality, while maximizing yield. The variety of 'Momotaro' was grown in the basic nutrient solution of 1.6 dS.m$^{[-10]}$ which was supplemented by three levels of seawater with EC 1.0, 2.0 or 3.0 dS.m$^{[-10]}$ . Tomato plants were cultivated in cool seasons. Plant growth characteristics were compared between treatments, and fruits were classified to analyse fruit quality characteristics according to ripening stages： MG, Br, Br＋3, Br＋5, Br＋7 and Br＋10. Adding seawater generally did not affect the shoot growth parameters such as plant height, leaf length, leaf width, internode length and chlorophyll content. Adding seawater negatively affected yield parameters such as the height and weight of fruit, marketable fruit weight per plant and marketable fruit yield. Therefore, the more yield reduction was obtained with the increasing level of seawater treatment. Fruit quality was improved by seawater treatment. The degree of the effect for $^{\circ}$Bx degree and sugars were the highest with the EC of seawater 2.0~3.0 dS.m$^{[-10]}$ , and at the Br＋5~Br＋7 of ripening stages. The relative abundance of tomato flavor, volatile components, was not generally affected by the seawater treatment with an exception of 6-methyl-5-hepten-2-one. The relative abundance of most volatile components increased as ripening progressed. The increment began at the Br stage and showed the highest increment at the Br＋5~Br＋7 stages. The results from these experiments suggest that seawater treatment of EC 3.6 dS.m$^{[-10]}$ for hydroponics is good for improving tomato quality. Fruit quality is the best at the Br＋5~Br＋7 ripening stages. It is considered that these results may be applied far use in hydroponic culture to improve fruit quality with minimum yield reduction.

• Applied Biological Chemistry
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• v.38 no.5
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• pp.468-472
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• 1995

Substantial amount of caryophyllene oxide (CPO) is present in the essential oils of traditionally-used folk medicinal plants and herbal spices. The CPO, produced via chemical and/or enzymatic reaction of caryophyllene (CP), has largely being used as a flavoring component and exhibited a variety of biological activities. Now, we report the antimutagenic activity of CPO determined by Ames's preincubation test. S-9 fraction was prepared from the liver of rats treated with Arochor 1254. Anatoxin $B_1\;(AFB_1)$ and 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) were used as mutagens. Reduction of mutagenicity of $AFB_1$ or IQ for S. typhimurium TA98 and TA100 by CPO was found to be a dose-dependant manner. CPO (500 ${\mu}g/plate$) reduced mutagenicity of AEB1 for S. typhimurium TA98 and TA100 to 89% and 71%, respectively. For IQ, similar results were observed against S. typhimurium TA98 and TA100, resulting in the inhibition percentage of 77% and 51%, respectively. CP also reduced mutagenicity of AEB1 and IQ for S. typhimurium TA98 and TA100, but the reduction rate was somewhat lowered relative to that of CPO. These results indicate that CPO could be developed as a potent antimutagenic flavoring agent.

• Journal of the Korean Academy of Child and Adolescent Psychiatry
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• v.5 no.1
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• pp.83-92
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• 1994

Paroxetine is a potent and selective serotoin re-uptake inhibitor. It is well known as an effective and safe antidepressant and increasingly used for neurotic or non-psychotic depression with anxiety symptoms. The present study assessed antidepressant and antianxiety efficacy and tolerability of paroxetine against placebo in child-adolescent and adult depressive neurosis patients. 232 subjects aged 8-55 years and meeting DSM-III-R criteria for depressive neurosis or dysthymia were divided into 8 subgroups according to their sex and age(8-11 yeard old, 12-17 years old, 18-35 years old and 36-55 years old subgroup in each male and female group). In each subgroup, the randomly assigned half of the patients were treated with paroxetine(10-30mg/day) and the others with placebo for the first 2 weeks in double blind fashion. After 1 week of drug-washout period, paroxetine and placebo groups were crossed over. The depression and anxiety symptoms were assessed with Hamilton Depression Scale(HDS) and Hamilton Anxiety Scale(HAS) at baseline and every 1 week during the trial periods. The levels of reduction in HDS and HAS scores from baseline after 2-week trial were compared between paroxetine- and placebo- treated periods by paired t-test. In all the 8 subgroups, statistically significant differences between paroxetine and placebo were found on the antidepressant efficacy after 2-week treatment. The antidepressant efficacy of paroxetine compared to placebo was most prominent in child and adolescent female groups. On anxiety symptoms, paroxetine was also significantly more effective than placebo. The antianxiety efficacy of paroxetine compared to placebo was most prominent in male and female child groups and young adult female group aged 18-35 years. As for the adverse effects of paroxetine, 3 out of 232 subjects reported mild indigestion and abdominal pain. however, in all the 3 cases, the symptoms improved without reduction of dosage or discontinuation of the drug. In conclusion, paroxetine showed significantly higher antidepressant and antianxiety efficacy compared to placebo in child-adolescent and adult depressive neurosis patients after 2-week treatment. Further trials of paroxetine in depressive neurosis are warranted to elucidate the long-term antidepressant and antianxiety efficacy of paroxetine.

• Journal of Sasang Constitutional Medicine
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• v.27 no.2
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• pp.270-287
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• 2015

Objectives To evaluate the neuroprotective effects of modified Yuldahanso-tang (MYH) in a Parkinson's disease mouse model. Methods 1) Four groups (each of 8 rats per group) were used in this study. 2) The neuroprotective effect of MYH was examined in a Parkinson's disease mouse model. C57BL/6 mice treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP, 30 mg/kg/day), intraperitoneal (i.p.) for 5 days. 3) The brains of 2 mice per group were removed and frozen at $-20^{\circ}C$, and the striatum-substantia nigra part was seperated. The protein volume was measured by Bradford method following Bio-Rad protein analyzing kit. Using mouse/Rat Dopamine ELISA Assay Kit. 4) The brains of 2 mice per group were separated and removed. TH-immunohistochemical was examined in the MPTP-induced Parkinson's disease mice to evaluate the neuroprotective effects of MYH on ST and SNpc. 5) Two mice out of each group were anesthetized and skulls were opened from occipital to frontal direction to take out the brains. The brains added TTC solution for 20 minutes for staining. 6) The water tank used for morris water maze test was filled with $28^{\circ}C$ water, and a round platform of 10cm in diameter was installed for mice to step on. The study was carried out once a day within 30 seconds, keep exercising to step on the platform in the pool. 7) The brains of two mice out of each group were fixed in 10% formaldehyde solution and paraphillin substance was infiltrated. They were fragmented by microtome, and observed under an optical microscope after Hematoxylin & Eosin staining. 8) A round acrylic cylinder with its upper side open was filled with clean water and depressive mouse models were forced to swim for 15 minutes. After 24 hours the animals were put in the same equipment for 5 minutes and were forced to swim. 9) The convenient, simple, and accurate high-performance liquid chromatography (HPLC) method was established for simultaneous determination of Neurotransmitters in MPTP-MYH group. Results 1) MYH possess Dopamine cell protective effect on MPTP-induced injury in striatum and substantia nigra pars compacta. 2) MYH inhibits the loss of tyrosine hydroxylase-immunoreacitive (TH-IR) cells in the striatum and substantia nigra pars compacta on MPTP-induced injury in C57BL/6 mice. 3) MYH possesses improvement effect on MPTP-induced memory deterioration in C57BL/6 mice through the reduction of prolongated Sort of lost time by MPTP injection using the Morris water maze test. 4) MYH possesses hippocampal neuron protective effect on MPTP-induced injury in C57BL/6 mice. 5) MYH possesses improvement effect on MPTP-induced motor behaviour deficits and depression in C57BL/6 mice through the reduction of prolongated losing motion by MPTP injection using the Forced swimming test. 6) MYH increases serotonin product amount on MPTP-induced injury in C57BL/6 mice. Conclusions This experiment suggests that the neuroprotective effect of MYH is mediated by the increase in Dopamin, TH-ir cell, Hippocampus and Serotonin. Furthermore, MYH essential oil may serve as a potential preventive or therapeutic agent regarding Parkinson's disease.

• Journal of Life Science
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• v.32 no.1
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• pp.11-22
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• 2022

N-methyl-D-aspartate (NMDA) receptors have received considerable attention regarding their involvement in glutamate-induced neuronal excitotoxicity. Resveratrol has been shown to exhibit neuroprotective effects against this kind of overactivation, but the underlying cellular mechanisms are not yet clearly understood. In this study, HT-22 neuronal cells were treated with NMDA in Mg²⁺-free buffer and subsequently used as an experimental model of glutamate excitotoxicity to elucidate the mechanisms of resveratrol-induced neuroprotection. We found that NMDA treatment causes a drop in MTT reduction ability, disrupts inside-negative transmembrane potential of mitochondria, depletes cellular ATP levels, and stimulates intracellular ROS production. Double fluorescence imaging studies demonstrated an increased formation of mitochondrial permeability transition (MPT) pores accompanied by apoptotic cell death, while cobalt protoporphyrin and bilirubin showed protective effects against NMDA-induced mitochondrial injury. On the other hand, zinc protoporphyrin IX significantly attenuated the protective effects of resveratrol which was itself shown to enhance heme oxygenase-1 (HO-1) mRNA and protein expression levels. In cells transfected with HO-1 small interfering RNA, resveratrol failed to suppress the NMDA-induced effects on MTT reduction ability and MPT pore formation. The present study suggests that resveratrol may prevent mitochondrial injury in NMDA- treated HT-22 cells and that enhanced expression of HO-1 is involved in the underlying cellular mechanism.

• Korean Journal of Soil Science and Fertilizer
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• v.10 no.1
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• pp.55-60
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• 1977

In order to study the availability of soil P and applied P to rice plant under water-logged system, a pot experiment with five soils having different levels of available P (24, 64, 100, 144 and 231 ppm) under four levels of applied P (0, 30, 60 and 90 ppm) was conducted. The availability of P was measured in terms of plant performance and the behaviors of P in soils were studied through the fractionation of various forms of P and by measuring the adsorption and desorption characteristics. The results are summarized us following. 1. The rice plant responded to applied P in the soils containing less than 144 ppm of available P as measured by Lancaster's method in terms of number of tillers in early growth stages. However, when the response of rice plant to applied P was evaluated in terms of grain yield there was no response even in the soil containing 24 ppm of available P. 2. Applied P was fixed as Al-P at the early stages and converted into Fe-P at later stages. 3. The P adsorption maxima of soils measured by Langmuir's isotherm ranged from 70 to 100mg/100g. No relationships between the level of available P and P adsorption maxima were observed. 4. There was a trend that the higher the level of available P, the higher the release of water soluble p. 5. The reduction of soil increased the level of available P by the factor of 1.8 times of air dried soils.

• Nutritional Sciences
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• v.6 no.4
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• pp.223-228
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• 2003

This study was carried out to investigate the effects of vitamin E on the cadmium contents of bone and on the calcium and phosphorous contents of the blood, urine and feces. Male Sprague-Dawley rats weighing 100$\pm$10g were randomly assigned to one normal group and three cadmium poisoned groups. The cadmium poisoned groups consisted of a vitamin E free diet (Cd-0E) group; a 40 mg vitamin E /kg diet (Cd-40E) group; and a 400 mg/kg diet (Cd-400E) group. Experimental animals were maintained on their respective diets for 20 weeks and were simultaneously administered 50 ppm $Cd^{2+}$ dissolved in the drinking water. At the end of the trial, the average hematocrit value in the Cd-0E group was 28.13% lower than in the normal group. However, the average hematocrit value in the Cd-400E group was significantly higher than in the Cd-0E and Cd-40E groups. WBC levels in the cadmium-poisoned groups were lower than in the normal group, but Cd-400E group levels were significantly higher than in the Cd-0E and Cd-40E groups. The contents of calcium of tibia has no significant difference between normal group and cadmium exposed group at $10^{th}$ week After 20 weeks, the calcium contents of the tibia in the Cd-0E and Cd-40E groups were lower than in the normal group by 25.5% and 22.1 %, respectively, although the calcium contents of the tibia in the Cd-400E group were higher than in the normal group. After 10 weeks, the calcium contents of the femur in the Cd-0E and Cd-40E groups were 19.25% and 15.45% lower than in the normal group, respectively, but the calcium contents of the femur in the Cd-400E group were at the same levels as in the normal group. The levels of calcium in the femur after 20 weeks were similar to the 10-week levels. Calcium levels of the urine in the Cd-0E and Cd-40E groups were 3.92 fold and 2.92 fold higher, respectively, than in the normal group, but levels in the Cd-400E group were significantly lower than in either the Cd-0E group or the Cd-40E group. Calcium levels of the feces in cadmium-poisoned groups were significantly higher than in the normal group, although levels in the Cd-400E group were significantly lower than in the Cd-0E and Cd-40E groups. Phosphorous levels of the blood in the Cd-0E group were 17% lower than in the normal group, although levels in the Cd-400E group were significantly higher than in the Cd-0E group. Phosphorous levels of the urine in the Cd-0E and Cd-40E groups were significantly higher than in the normal group, while Cd-400E group levels were found to be at the same level as in the normal group. Cadmium contents of the tibia in the Cd-40E and Cd-400E groups were 13% and 17% lower, respectively, than in the Cd-0E group. Regarding cadmium levels in the femur, only the Cd-400E group achieved lower levels (10% lower) than the Cd-0E group. In conclusion, vitamin E supplementation resulted in a suppression of the release of calcium from bone, and a reduction in the excretion of calcium via the urine and feces, thus having a normalizing effect on calcium metabolism in rats with chronic cadmium poisoning.

• Korean Journal of Food Science and Technology
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• v.50 no.4
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• pp.420-429
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• 2018

Skin ageing is associated with compromised performance of its fundamental barrier functions, with undesirable changes in appearance. Since this may introduce a detrimental impact on the quality of life, significant effort to discover effective ingredients against ageing is being invested. Recently, collagen hydrolysates containing tripeptides such as GlyPro-Hyp (GPH) have been developed with anticipation of improved effects compared to that of existing collagen hydrolysate-products. To evaluate the cutaneous hydration effect of collagen tripeptides (CTP), meaningful biomarkers in human dermal fibroblasts (HDF) and NC/Nga Tnd mice were analyzed in this study. Increased levels of ceramide kinase, hyaluronic acid, collagen 1A, and hyaluronan synthase-2 (HAS2), and decreased levels of hyaluronidase-1 (HYAL1) and CD44 in HDF cells were demonstrated. Furthermore, significant reduction of transepidermal water loss (TEWL), scratching behavior, HYAL1, $TNF-{\alpha}$ and IL-6 and increased water content and HAS2 were verified by in vivo tests. These results strongly suggest the potential of CTP as a skin hydration agent.

• Korean journal of food and cookery science
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• v.29 no.4
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• pp.417-424
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• 2013

This study was carried out to investigate the effect of safflower seed on the physicochemical properties of ground pork during frozen storage. Three types of ground pork were evaluated: 20% pork back fat added (T0, control), 10% pork back fat and 10% added safflower seed (T1), and 20% added safflower seed (T2). Water holding capacity decreased with longer storage period, and that of T2 was the highest (p<0.05). Cooking loss increased with longer storage period, and that of T1 and T2 was higher than that of T0 (p<0.05). The reduction in diameter of T0 increased, but that of T1 and T2 was not significantly different with longer storage period. Hardness and chewiness increased, but springiness decreased with longer storage period (p<0.05). Hardness, springiness and chewiness of T2 was the highest (p<0.05). The pH decreased with longer storage period (p<0.05), and those of T0, T1 and T2 were pH 5.41, 5.43 and 5.32, respectively, after 50 days of storage. The TBARS (2-thiobarbituric acid reactive substances) values of T0 and T1 increased, but that of T2 was not significantly different with longer storage period. The TBARS values of T0, T1 and T2 were 4.76, 2.77 and 0.54 mg malonaldehyde/kg, respectively, after 50 days of storage. The $L^*$, $a^*$ and $b^*$ value of T0 was the highest among the samples (p<0.05), the $a^*$ value of the samples decreased with longer storage period (p<0.05). In conclusion, this study demonstrates that the addition safflower seed tended to improve physiological properties and antioxidative effects.