• Journal of Plant Biotechnology
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• v.46 no.3
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• pp.165-171
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• 2019

Marker-assisted backcrossing (MABC) is useful for selecting an offspring with a highly recovered genetic background for a recurrent parent at early generation to various crops. Moreover, marker-assisted backcrossing (MABC) along with marker-assisted selection (MAS) contributes immensely to overcome the main limitation of the conventional breeding and it accelerates recurrent parent genome (RPG) recovery. In this study, we were employed to incorporate rin gene(s) from the donor parent T13-1084, into the genetic background of HK13-1151, a popular high-yielding tomato elite inbred line that is a pink color fruit, in order to develop a rin HK13-1084 improved line. The recurrent parent genome recovery was analyzed in early generations of backcrossing using SNP markers obtained from genotyping-by-sequencing analysis. From the $BC_1F_1$ and $BC_2F_1$ plants, 3,086 and 4868 polymorphic SNP markers were obtained via GBS analysis, respectively. These markers were present in all twelve chromosomes. The background analysis revealed that the extent of RPG recovery ranged from 56.7% to 84.5% and from 87.8% to 97.8% in $BC_1F_1$ and $BC_2F_1$ generations, respectively. In this study, No 5-1 with 97.8% RPG recovery rate among $BC_2F_1$ plants was similar to HK13-1151 strain in the fruit shape. Therefore, the selected plants were fixed in $BC_2F_2$ generation through selfing. MAS allowed identification of the plants that are more similar to the recurrent parent for the loci evaluated in the backcross generations. MABC can greatly reduce breeding time as compared to the conventional backcross breeding. For instance, MABC approach greatly shortened breeding time in tomato.

• Journal of Life Science
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• v.29 no.1
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• pp.76-83
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• 2019

In this study, the optimal growth and poly(3-hydroxybutyrate) (PHB) biosynthesis of Pseudomonas sp. EML2 were established using waste frying oil (WFO) as a cheap carbon source. The fatty acid composition of WFO and fresh frying oil (FFO) were analyzed by gas chromatography. The unsaturated and saturated fatty acid contents of the FFO were 82.6% and 14.9%, respectively. These contents changed in the WFO. The compositional change in the unsaturated fatty acid content in the WFO was due to a change in its chemical and physical properties resulting from heating, an oxidation reaction, and hydrolysis. The maximum dry cell weight (DCW) and PHB yield (g/l) of the isolated strain Pseudomonas sp. EML2 were confirmed under the following culture conditions: 30 g/l of WFO, 0.5 gl of $NH_4Cl$, pH 7, and $20^{\circ}C$. Based on this, the growth and PHB yield of Pseudomonas sp. EML2 were confirmed by 3 l jar fermentation. After the cells were cultured in 30 g/l of WFO for 96 h, the DCW, PHB content, and PHB yield of Pseudomonas sp. EML2 were 3.6 g/l, 73 wt%, and 2.6 g/l, respectively. Similar results were obtained using 30 g/l of FFO as a carbon source control. Using the FFO, the DCW, PHB content, and PHB yield were 3.4 g/l, 70 wt%, and 2.4 g/l, respectively. Pseudomonas sp. EML2 and WFO may be a new candidate and substrate, respectively, for industrial production of PHB.

• Journal of Life Science
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• v.31 no.1
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• pp.90-99
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• 2021

In this study, to reduce the production cost of poly(3-hydroxyalkanoates) (PHA), optimal cell growth and PHA biosynthesis conditions of the isolated strain Pseudomonas sp. EML8 were established using waste frying oil (WFO) as the cheap carbon source. Gas chromatography (GC) and GC mass spectrometry analysis of the medium-chain-length PHA (mcl-PHAWFO) obtained by Pseudomonas sp. EML8 of WFO indicated that it was composed of 7.28 mol% 3-hydrxoyhexanoate, 39.04 mol% 3-hydroxyoctanoate, 37.11 mol% 3-hydroxydecanoate, and 16.58 mol% 3-hydroxvdodecanoate monomers. When Pseudomonas sp. EML8 were culture in flask, the maximum dry cell weight (DCW) and the mcl-PHAWFO yield (g/l) were showed under WFO (20 g/l), (NH4)2SO4 (0.5 g/l), pH 7, and 25℃ culture conditions. Based on this, the highest DCW, mcl-PHAWFO content, and mcl-PHAWFO yield from 3-l-jar fermentation was obtained after 48 hr. Similar results were obtained using 20 g/l of fresh frying oil (FFO) as a control carbon source. In this case, the DCW, the mcl-PHAFFO content, and the mcl-PHAFFO yields were 2.7 g/l, 62 wt%, and 1.6 g/l, respectively. Gel permeation chromatography analysis confirmed the average molecular weight of the mcl-PHAWFO and mcl-PHAFFO to be between 165-175 kDa. Thermogravimetric analysis showed decomposition temperature values of 260℃ and 274.7℃ for mcl-PHAWFO and mcl-PHAFFO, respectively. In conclusion, Pseudomonas sp. EML8 and WFO could be suggested as a new candidate and substrate for the industrial production of PHA.

• Microbiology and Biotechnology Letters
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• v.49 no.4
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• pp.552-558
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• 2021

Traditional foods are manufactured according to the characteristics of each region for the nations of the world. Korea has mainly farmed, and seasonings have developed around rice and vegetables. In fermented foods, lactic acid bacteria such as Lactobacillus sp. and Pediococcus sp. and Bacillus sp. were isolated and identified from fermented foods. In this study, lactic acid bacteria were isolated and identified from commercially available traditional Korean fermented foods, and candidate strains were selected through antibacterial activity tests on human and fish disease bacteria. Thereafter, the final strain was selected by examining the resistance to simulated gastric and intestinal fluids, and hemolysis. The three (M1, K1, C13) final selected latic acid bacteria were miced with prebiotics and the antibacterial activity of synbiotics was evaluated. As for the fist antibacterial activity result, C13 showed high antibacterial acitivity in human diseases and fish diseases. Then, M1, K1 and C13, which did not produce β-haemolysis and were resistant to simulated gastric and intestinal fluids, were subjected to the second antibacterial activity of synbiotics. When the three prebiotics (FOS, GOS, Inulin) and probiotics (M1, K1, C13) were mixed, the antibacterial activity was increased or inhibited. Based on the 16S rRNA gene sequencing results, K1 and M1 were analyzed as Bacillus tequiensis 99.72%, Bacillus subtilis 99.65%, Bacillus inaquosorum 99.72%, Bacillus cabrialesii 99.72%, Bacillus stercoris 99.58%, Bacillus spizizenii 99.58%, Bacillus halotolerans 99.58%, and Bacillus mojavensis 99.51%. And C13 was analyzed as Bacillus velezensis 99.71%, Bacillus nematocida 99.36%, Bacillus amyloliquefaciens 99.44%, Bacillus atrophaeus 99.22%, and Bacillus nakamurai 99.44%.

• Journal of the korean academy of Pediatric Dentistry
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• v.50 no.2
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• pp.168-178
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• 2023

Objectives: This study aimed to compare the physical properties and antibacterial effectiveness of iodoform based root filling pastes, Vitapex^® and Metapex^®, with sodium iodide root filling paste (NaI paste) for primary teeth. Materials and Methods: The physical properties (flowability, film thickness, radiopacity) of the pastes were evaluated according to ISO 6876:2012. The antibacterial activity against Enterococcus faecalis strain (ATCC 6538) was evaluated using a direct contact test. Results: There was no significant statistical difference (p > 0.05) observed in the flow and film thickness of NaI paste when compared to the currently available root canal filling materials. The average flow capacities for Vitapex^®, Metapex^®, and NaI paste were 15.40 mm, 21.25 mm, and 20.01 mm, respectively. The average film thickness for Vitapex^®, Metapex^®, and NaI paste were 33.3 ㎕, 22.6 ㎕, and 25.0 ㎕, respectively. However, NaI paste showed lower radiopacity than the existing materials, and this difference was statistically significant (p < 0.05) NaI paste demonstrated higher antimicrobial activity than the available materials, and this difference was also statistically significant (p < 0.05). Conclusion: Compared to the existing commercialized root canal filling materials, NaI paste exhibited similar performance in terms of flow and film thickness, and superior antimicrobial activity against E. faecalis. Hence, NaI paste could be a promising root filling material for primary teeth and may be a potential alternative to existing materials.

• Journal of Life Science
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• v.34 no.7
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• pp.465-475
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• 2024

Lactiplantibacillus plantarum K9 is a probiotic strain that can be utilized from various bioactive substances isolated from Protaetia brevitarsis seulensis larvae. In this study, a genetic analysis of L. plantarum K9 revealed the existence of a bacterial chromosome and three plasmids. The glycolysis pathway and pentose phosphate pathway were examined for their normal functioning via an analysis of the core metabolic pathways of L. plantarum K9. Since the key enzymes, fluctose-1,6-bisphospatase (EC: 3.1.3.11) and 6-phosphogluconate dehydratase (EC: 4.2.1.12)/2-keto-deoxy-6-phosphogluconate (KDPG) aldolase (EC: 4.2.1.55), of gluconeogenesis and the ED pathway were not identified from the L. plantarum K9 genome, we suggest that gluconeogenesis and the ED pathway are not performed in L. plantarum K9. Additionally, while some enzymes, related to fumarate and malate biosyntheses, involved in the TCA cycle were identified from L. plantarum K9, the enzymes associated with the remaining TCA cycle were absent, indicating that the TCA cycle cannot proceed. Meanwhile, based on our findings, we propose that the oxidative electron transport system performs class IIB-type (bd-type) electron transfer. In summary, we assert that L. plantarum K9 performs homolactic fermentation, executes gluconeogenesis and the pentose phosphate pathway, and carries out energy metabolism through the class IIB-type oxidative electron transport system. Therefore, we suggest that L. plantarum K9 has relatively high lactic acid production, and that it has excellent antibacterial activity, as a result, compared to other lactic acid bacterial strains. Moreover, we speculate that L. plantarum K9 has an oxidative electron transport capability, indicating that it is highly resistant to oxygen and suggesting that it has fine cultivation characteristics, which collectively make it highly suitable for use as a probiotic.

• Journal of Biosystems Engineering
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• v.1 no.1
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• pp.15-15
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• 1976

In analyzing the operational characteristics of a rice whitening machine, the internal radial pressure of the machine was measured using strain gage equipment. Changes in cylinder and feed screw configurations, screen type, cylinder speed and counter-pressure levels were examined to determine their impact on the quality and quantity of milled rice and the performance of the machine. The results are summarized as follows: 1. The internal radial pressure in the whitening chamber varied with the surface condition of the grain being processed. During the first or second pass through the machine, pressure was relatively low, reached a maximum after two to three passes with combinations I and II, three to six with combination III and then began to fall. 2. The pitch of the feed screw and the size of the feed gate opening which determine the rate of entry of grain into the whitening chamber, appeared to be the most important factor aff-::cting the degree of radial pressure, quality and quantity of milled rice and the efficiency of the machine. Using a feed screw with a wide pitch (4.8cm), radial pressure was relatively high and head rice recovery ratio \vere quite low. In this case capacity and machine effic?iency were much higher than obtained when using a feed screw with a narrow pitch (2.3cm). Very significant responses in radial pressure, head rice recovery rates and machine capacity were observed with changes in cylinder speed and counter-pressure levels when using the wide pitch feed screw. 3. The characteristics of the screen which surrounds the whitening chamber had an important effect on whitening efficiency. The existence of small protuberances on the original screen resulted in significant increases in both machine capacity and efficiency but without a significant decrease in head rice recovery or development of excessive radial pressure. Further work is required to determine the effects of screen surface conditions and the shape of the cylinderical steel roller on the rate of bran removal, machine efficiency and recovery rates. The size of the slotted perforations 0:1 the screen affects total milled rice recovery. The opening size on the original screen was fabricated to accommodate the round shape of Japonica rice varieties but was not suitable for the more slender Indica type. Milling Indica varieties with this screen resulted in a reduction in total milled rice recovery. 4. An increase in cylinder speed from 380 to 820 rpm produced a positive effect on head rice recovery for all machine combinations at every level of counter-pressure used in the tests. Head rice recovery was considerably lower at 380rpm using a wide screw pitch when compared to the results obtained at speeds from 600 to 820 r.p.m. The effects of cylinder speed On radial pressure, capacity and machine efficiency showed contrasting results, depending on the width of the feed screw pitch. With a narrow feed screw pitch (2.3cm), a direct proportional relationship was observed bet?ween cylinder speed and both radial pressure and machine efficiency. In contrast, using a 4.8 centimeter pitch feed roller produced a series of inverse relationships between the above variables. Based on the results of this study it is recommended when milling Indica type long grain rice varieties that the cylinder speed of the original machine be increased from 500-600 rmp up to a minimum of 800 rpm to obtain a greater abrasive effect between the grain and the screen. The pitch of the feed screw should be also reduced to decr?ease the level of internal radial pressure and to obtain higher machine efficiency and increased quality of milled rice with increased cylinder speeds. Further study on the interaction between cylinder speed and feed screw pitch is recommended. 5. An increase in the counter pressure level produced a negative effect On the head rice recovery with an increase in radial pressure, capacity, and machine efficiency over all combinations and at every level of cylinder speed. 6. Head rice recovery rates were conditioned primarily by the pressure inside the whitening chamber. According to the empirical cha racteristics curve developed in this study, the relationships of head rice recovery ($Y_h$) and machine capacity ($Y_c$/TEX>) to internal radial pressure ($X_p$) followed an inverse quadratic function and a linear function respectively: $$Y_h^\Delta=\frac{1}{{1.4383-0.2951X_p^\ast+0.1425X_p^{\ast\ast}}^2} , (R^2=0.98)$$$$Y_c^\Delta=-305.83+374.37X_p^{\ast\ast}, (R^2=0.88)$$The correlation between capacity and power consumption per unit of brown rice expressed in the following exponential function: $$Y_c^\Delta=1.63Y_c^{-0.7786^\{\ast\ast}, (R^2=0.94)$$These relationships indicate that when radial pressure increases above a certain range (1. 6 to 2.0 kg/$cm^2$ based On the results of the experiment) head ricerecovery decrea?ses in a quadratic relation with a inear increase in capacity but without any decrease in power consump tion per unit of brown rice. On the other hand, if radial pressure is below the range shown above, power consumption increases dramatically with a lin?ear decrease in capacity but without significant increases in head rice recovery. During the operation of a given whitening machine, the optimum radial pressure range or the correct capacity range should be selected by controlling the feed rate and/or counter-pressure keeping in mind the condition of the grain, particulary the hardness. It was observed that the total number of passes is related to radial pessure level, feed rate and counter-pressure level. The higher theradial pressure the fewer num?ber of pass required but with decreased head rice recovery. In particular, when using high feed rates, the total number of passes should be increased to more than three by reducing the counter-pressure level to avoid decreaseases in head rice recovery (less than 65 percent head rice recovery on the basis of brown rice) at every cylinder speed. 7. A rapid rise in grain temperature seemed to have a close relationship with the pressure generated inside the whitening chamber and, subsequently with head rice reco?very rates. The higher the rate of increase, the lower were the resulting head rice recoveries.

• Journal of Biosystems Engineering
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• v.1 no.1
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• pp.17-48
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• 1976

In analyzing the operational characteristics of a rice whitening machine, the internal radial pressure of the machine was measured using strain gage equipment. Changes in cylinder and feed screw configurations, screen type, cylinder speed and counter-pressure levels were examined to determine their impact on the quality and quantity of milled rice and the performance of the machine. The results are summarized as follows: 1. The internal radial pressure in the whitening chamber varied with the surface condition of the grain being processed. During the first or second pass through the machine, pressure was relatively low, reached a maximum after two to three passes with combinations I and II, three to six with combination III and then began to fall. 2. The pitch of the feed screw and the size of the feed gate opening which determine the rate of entry of grain into the whitening chamber, appeared to be the most important factor aff-::cting the degree of radial pressure, quality and quantity of milled rice and the efficiency of the machine. Using a feed screw with a wide pitch (4.8cm), radial pressure was relatively high and head rice recovery ratio \vere quite low. In this case capacity and machine effic\ulcorneriency were much higher than obtained when using a feed screw with a narrow pitch (2.3cm). Very significant responses in radial pressure, head rice recovery rates and machine capacity were observed with changes in cylinder speed and counter-pressure levels when using the wide pitch feed screw. 3. The characteristics of the screen which surrounds the whitening chamber had an important effect on whitening efficiency. The existence of small protuberances on the original screen resulted in significant increases in both machine capacity and efficiency but without a significant decrease in head rice recovery or development of excessive radial pressure. Further work is required to determine the effects of screen surface conditions and the shape of the cylinderical steel roller on the rate of bran removal, machine efficiency and recovery rates. The size of the slotted perforations 0:1 the screen affects total milled rice recovery. The opening size on the original screen was fabricated to accommodate the round shape of Japonica rice varieties but was not suitable for the more slender Indica type. Milling Indica varieties with this screen resulted in a reduction in total milled rice recovery. 4. An increase in cylinder speed from 380 to 820 rpm produced a positive effect on head rice recovery for all machine combinations at every level of counter-pressure used in the tests. Head rice recovery was considerably lower at 380rpm using a wide screw pitch when compared to the results obtained at speeds from 600 to 820 r.p.m. The effects of cylinder speed On radial pressure, capacity and machine efficiency showed contrasting results, depending on the width of the feed screw pitch. With a narrow feed screw pitch (2.3cm), a direct proportional relationship was observed bet\ulcornerween cylinder speed and both radial pressure and machine efficiency. In contrast, using a 4.8 centimeter pitch feed roller produced a series of inverse relationships between the above variables. Based on the results of this study it is recommended when milling Indica type long grain rice varieties that the cylinder speed of the original machine be increased from 500-600 rmp up to a minimum of 800 rpm to obtain a greater abrasive effect between the grain and the screen. The pitch of the feed screw should be also reduced to decr\ulcornerease the level of internal radial pressure and to obtain higher machine efficiency and increased quality of milled rice with increased cylinder speeds. Further study on the interaction between cylinder speed and feed screw pitch is recommended. 5. An increase in the counter pressure level produced a negative effect On the head rice recovery with an increase in radial pressure, capacity, and machine efficiency over all combinations and at every level of cylinder speed. 6. Head rice recovery rates were conditioned primarily by the pressure inside the whitening chamber. According to the empirical cha racteristics curve developed in this study, the relationships of head rice recovery ($Y_h$) and machine capacity ($Y_c$/TEX>) to internal radial pressure ($X_p$) followed an inverse quadratic function and a linear function respectively: $$Y_h^\Delta=\frac{1}{{1.4383-0.2951X_p^\ast+0.1425X_p^{\ast\ast}}^2} , (R^2=0.98)$$ $$Y_c^\Delta=-305.83+374.37X_p^{\ast\ast}, (R^2=0.88)$$ The correlation between capacity and power consumption per unit of brown rice expressed in the following exponential function: $$Y_c^\Delta=1.63Y_c^{-0.7786^\{\ast\ast}, (R^2=0.94)$$ These relationships indicate that when radial pressure increases above a certain range (1. 6 to 2.0 kg/$cm^2$ based On the results of the experiment) head ricerecovery decrea\ulcornerses in a quadratic relation with a inear increase in capacity but without any decrease in power consump tion per unit of brown rice. On the other hand, if radial pressure is below the range shown above, power consumption increases dramatically with a lin\ulcornerear decrease in capacity but without significant increases in head rice recovery. During the operation of a given whitening machine, the optimum radial pressure range or the correct capacity range should be selected by controlling the feed rate and/or counter-pressure keeping in mind the condition of the grain, particulary the hardness. It was observed that the total number of passes is related to radial pessure level, feed rate and counter-pressure level. The higher theradial pressure the fewer num\ulcornerber of pass required but with decreased head rice recovery. In particular, when using high feed rates, the total number of passes should be increased to more than three by reducing the counter-pressure level to avoid decreaseases in head rice recovery (less than 65 percent head rice recovery on the basis of brown rice) at every cylinder speed. 7. A rapid rise in grain temperature seemed to have a close relationship with the pressure generated inside the whitening chamber and, subsequently with head rice reco\ulcornervery rates. The higher the rate of increase, the lower were the resulting head rice recoveries.

• The Korean Journal of Mycology
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• v.21 no.3
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• pp.200-211
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• 1993

Somatic hybrids of Pleurotus florida ASI 2016 and Pleurotus ostreatus ASI 2018 were obtained by protoplast fusion. The 40 fusants($P1{\sim}P40$) was examined for the yield on fermented and pasteurized rice straw in a tray. The carpophore yield of them were showed as the range of $27.0{\sim}155.2$, based on parental values of 100(ASI 2018), The pilei of fusants between orange white colored P. florida and dark grey colored P. ostreatus had mixed colors in the young stage. Other breeding programmes were performed to improve new varieties with high yield and good quality. A new oyster mushroom variety, Wonhyeongneutaribeosus(P72), was developed at the Agricultural Sciences Institute, Rural Development Administration in 1990. This P.florida-ostreatus-ostreatus hybrid P72 was selected from 38 protoplast fusion products($P41{\sim}P78$) between P.florida-ostreatus recombinant P5-M 43-arg rib and P. ostreatus ASI 2-13-0 2001-19-pro orn. The yield indexes of 38 hybrids ranged $40.5{\sim}152.7$ compared with the parental values of 100(ASI 2001). Hybrid P72 was characterized by the large fruiting bundle of semispherical shape with long stipe and by the small and circular pileus, resulting in lower harvesting cost. A significant increase in carpophore production was observed in somatic hybrids of protoplasts due to heterosis. A comparision of hybrid with parents P72 was made using isozyme analysis. The esterase banding patterns could be characterized by new bands in the hybrids. Seven fusion products of four crosses between P.florida ASI 2016 and P. ostreatus ASI 2018 were analysed with respect to the distribution of progenies and segregation of gene markers by random basidiospore analysis. Segregation of alleles should yield progeny of four genotypes in a Mendelian ratio of 1 : 1 : 1 : 1 for prototrophs, auxotrophs of one parental type, auxotrophs of the other parental type, and auxotrophic recombinants, respectively. However, five fusants of them did not detect one parental, P.ostreatus, type. Basidiospores could yield progeny of 16 genotypes in the cross of one of the recombinant P5-M43-arg $rib{\times}P. ostreatus$ ASI 2-13-pro orn but the segregants of three fusants were not detected clearly. The allele ratio of loci could be expected 1 : 1 : 1 : 1 for arg, rib, pro and orn. The ratio, however, would be changed to 4 : 1 : 1 : 1 with increasing proportion of argo In almost all the fusants, prototrophic recombinants were recovered in large numbers against auxotrophic markers. Parental genotypes were recovered with the recombinant progeny amounting to $38.68{\sim}99.56%$. The analysis provides proof of heterokaryosis and strong evidence for haploidy of vegetative nuclei, a sexual cycle consisting of nuclear fusion and meiosis.

• Journal of Oral Medicine and Pain
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• v.31 no.3
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• pp.199-210
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• 2006

The epidemiological researches on the inpatients hospitalized at the oral medicine ward have been continuously carried out since 1970, and most researches have been performed by centering around the oral medicine wards of college hospitals. Numerous specialists have been produced after the establishment of oral medicine, and they have been active in various fields. As dental clinics have gotten bigger, the function of oral medicine in the secondary clinics is being brought out. As admissive channel, case features, case composition and otherwise have not been researched for a long time, the related researches should be carried out from now on. Hereupon, this study was carried out by targeting the 100 inpatients hospitalized at the oral medicine ward of Sun Hospital located in Daejeon Korea, through questionnaire. As the result, the following results were derived. 1. The ages of the inpatients in Sun Hospital were $29.21{\pm}11.31$ on the average; 71 females' mean average was $29.63{\pm}11.29$ and 29 males' mean average was $28.17{\pm}11.48$. In regard of school career, the patients who finished high-school course or higher accounted for 78%; the patients' school career seemed to be relatively high. The patients who complained of temporomandibular pain accounted for the highest proportion with 65%. In motivation to visit this hospital, internet surfing was 11%, mass media was 10%, acquaintance's introduction was 38%. The patients, who were hospitalized at another hospital due to the same symptom, accounted for 56%. The dental clinics, which made the patients visit this hospital, accounted for 20%. The patients, who were previously aware that the present symptom should be treated by oral medicine, accounted for 38%. The patients, who were not aware of the fact in advance, were 62%. The respondents of 51% answered that they were aware of the fact one month or below before hospitalization. 2. The patients, who complained of craniocervical ache, accounted for 58%; the patients, whose ache aches affect dailylife, were 22%. Continuous ache was 14% and intermittent ache was 68%, and dull pain was 23%. 3. Life variations were compared with each other by using SRRS (Social Readjustment Rating Scale). In consequence, the variation within 3 years indicated a significant difference in the both groups but the variation within 6 months did not indicate any differences. 4. In regard of the questionnaire on the incidents happened for a week, the ache-group was compared with the group free from the ache. As the result, the number of strain arisen for a week, the decrease of favorite works and sudden fear indicated a significant difference. Pleasant feeling and the decrease of interests in looks did not indicate a significant difference, but came close to the significance. 5. In the questionnaire on impatience, the ache-group indicated higher value but there was not a significant difference. 6. In the questionnaire on the symptoms caused by stress, the two groups indicated significant differences in the item of 'the teethridge itches and feels a tooth rising' and 'the occiput or the nape is stiff.' In the item 'the inside of the cheek or the teethridge are widely peeled off, accompanied with ache and hemorrhage', 'the face has acne or pimple' and 'headache frequently attacks', a significant difference was not observed but the two groups came close to the significance.