• Title/Summary/Keyword: Developmental stage

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Discontinuous Percoll Gradients Enrich X-Bearing Porcine Sperms and Female Embryos (불연속 Percoll 원심분리에 의한 돼지 X-정자와 자성배아에 관한 연구)

  • Shim, Dae-Yong;Yoo, Seong-Jin;Kang, Han-Seung;Yoo, Jeong-Min;Lee, Chae-Kwan;Kang, Sung-Goo
    • Development and Reproduction
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    • v.5 no.1
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    • pp.47-52
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    • 2001
  • Predetermination of sex in livestock of offpring is in great demand and is of critical importance to providing for the most efficient production of the animal ariculture. Such a sexing techlology would also enhance the economy of conventional artificial insemination(AI) and aid the porcine industry. The purpose of this study was to evaluate the efficiency of enriching X-bearing porcine sperm using discontinuous percoll gradients and PCR mefhod. Semen was collected from mature boars of proven fertility center (AI center KimHae). Sperm was leaded on the isotonic discontinuous percoll gradient and then it was centrifuged at 120 ${\times}$ g for 20 minutes. After centrifugation, sperm included in each fraction were recovered (7${\times}$10$^6$ sperms/ml) and then sperm genomic DNA was extractedfor the PCR. SRY gene was used to evaluate the ratio between X and Y sperm in the separated fractions. Ju viro ffrtilization wascarried out by adding the unseparated sperm (control) or separated (experimental poop) to the matured oocytes in TCM-199. Embryos for sex determination were obtained at 2 cell stage and then was used for SRY gene amplification. After centrifugation of discontinuous percoll gradient, the most motile sperm was obtained at 95% fiaction (94.4% ${\pm}$ 5.1%, p < 0.01). The PCR analysis evaluated that 30%, 50% and 65% fractions were Y sperm rich, whereas 80% and 95% fractions were X sperm rich. PCR analysis with each porcineembryo showed that 33.3% of control and 66.7% of experimental group were determined as female embryos. In conclusion, in vitro matured oocytes inseminated with sperms (95% fraction) prepared by percoll gradient centrifugation showed high fertilization rates and female embryos than control sperms.

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The Effect of Modified Cryopreservation Method on Viability of Frozen-thawed Blastodermal Cells on the Korean Native Chicken(Ogolgye Breed) (한국재래닭(오골계)종 배반엽세포에 있어서 동결 방법의 개선이 융해 후 생존율에 미치는 영향)

  • Kim, Hyun;Kim, Dong-Hun;Park, Soo-Bong;Choi, Seong-Bok;Ko, Yeoung-Gyu;Kim, Jae-Hwan;Do, Yoon-Jung;Park, Hae-Geum;Kim, Sung-Woo
    • Reproductive and Developmental Biology
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    • v.36 no.1
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    • pp.65-70
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    • 2012
  • For reconstituting genetic resource(Korean Native Chicken: KNC) with grem-line chimeric chicken made with cryopreserved biastdermal cells, the experiments were carried out to optimize cryopreservating conditions. Stage X biastdemal cells were collected from KNC embryos and dissociated. Cells were susupended in medium containing cyopretectant and fetal bovine serum(FBS), and distributed into plastic ampules. Cell susupensions were seeded to induce ice formation at $-7^{\circ}C$ to $-35^{\circ}C$ at in the experiments, the effect of modification of dissociation way, concentration of FBS and cell density on the vaibility of frezen-thawed cells were investigated by trypan blue exclusion. Then change the way of cell dissociation from pipetting to short time vortexing, viability of frozen-thawed cell tended to be increaced from 29 % to 52 %. Increase concentraition of FBS in frozen medium from 20 % to 80 % made viability of thawed cell from 28 % to 35 %. The viability of thawed cells were 33.9% frozen at 2 embryos/0.5 ml, and 43.6 % frozen at 20 embryos/0.5 ml. Furthermore, combination of three modifications make big improvement. The viability of frozen-thawed cell was 60 % for combinated method, and 41 % for general method. This result means the advance to practical cryoreservation of blastdermal cell of the KNC(Ogolgye breed).

Effects of Kinds and Concentrations of Cryoprotectants, PVP on Survival Rate of Vitrified Porcine Embryos (내동제의 종류와 농도, PVP 첨가가 돼지 수정란의 Vitrification 동결 융해 시 생존율에 미치는 영향)

  • Lim J. G.;Quan J. H.;Lee K. S.;Kim S. K.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.129-135
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    • 2005
  • The present study examines the effects of kinds and concentrations of cryoprotectants, PVP and sucrose and trehalose on the survival rate of vitrified porcine embryos. 1. The developmental stages for the embryos used in vitrification were $245(23.0\%)$ for 2 cell stage, $256(24.1\%)$ for the blastocyst, $234(22.0\%)$ for the early blastocyst $221(20.8\%)$ from the expanded blastocyst and $107(10.1\%)$ from hatching blastocyst out of 1,063 embryos. 2 The survival rate of morula, early blastocyst and expanded blastocyst vitrification-thawed with EDT and EGS were $69.1\%,\;70.3\%,\;69.8\%\;and\;62.5\%,\;61.7\%,\;63.6\%$, respectively. The expanded blastocyst treated with EDS showed the highest survival rate compared with the other cryoprotectants. 3. The survival rate of early blastocyst, expanded blastocyst and hatching blastocyst vitrification-thawed with EDS diluted in $medium + 10\%$ FCS were $61.1\%,\;27.8\%,\;16.7\%$, respectively. This result were love. than the control of group $(92.3\%,\;71.2\%,\; 55.8\%)$. 4. The survival rate of embryos vitrified with EDS and EDT supplemented with $10\%\;and\;20\%$ PVP were $74.3\%,\;77.5\%\;and\;79.4\%,\;71.1\%$, respectively. The survival rate of vitrified embryos cultured for $24\~48$ hours were $37.1\%,\; 40.0\%\;and\;35.3\%,\;31.6\%$ which were significantly lower than that of non-cultured embryos. The survival rate of embryos vitrified with EDS and EDT supplemented between $10\%\;or\;20\%$ PVP did not have a significant difference. 5. The survival rate of embryos vitrification-thawed with EDS to morula, early blastocyst, expanded blastocyst and hatching blastocyst were $58.2\%,\; 36.4\%,\;14.5\%$ to morula, $62.5\%,\;45.8\%,\;20.8\%$ to early blastocyst, $74.1\%,\;61.1\%,\;29.6\%$ to expanded blastocyst and $60.0\%,\;40.0\%,\;14.0\%$ to hatching blastocyst.

Analyses of CO2 Concentration and Balance in a Closed Production System for King Oyster Mushroom and Lettuce (밀폐형 식물생산시스템 내 새송이 버섯과 상추의 혼합 재배 비율에 따른 CO2 농도 변화 및 균형 분석)

  • Jung, Dae Ho;Kim, Chan Kyo;Oh, Kyung Hun;Lee, Dong-Hyeon;Kim, Minsu;Shin, Jong Hwa;Son, Jung Eek
    • Horticultural Science & Technology
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    • v.32 no.5
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    • pp.628-635
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    • 2014
  • The large amount of $CO_2$ emitted from mushrooms during incubation and developmental stages can be utilized in plant production systems as a $CO_2$ source. The objectives of this study were to measure the $CO_2$ emission and absorption rates of mushroom and lettuce, respectively, and to analyze the $CO_2$ concentrations at various ratios of mushroom and lettuce in a closed production system. The $CO_2$ emission rate of king oyster mushrooms (Pleurotus eryngii ( DC.) Qu$\acute{e}$l) and $CO_2$ absorption rate of lettuces (Lactuca sativa L. cv. Asia Heuk Romaine) were measured by using two closed acryl chambers ($1.0m{\times}0.8m{\times}0.5m$) in which indoor temperatures were maintained at $18^{\circ}C$ and $22^{\circ}C$, respectively. The lettuce was grown at a light intensity of PPF $340mol{\cdot}m^{-2}{\cdot}s^{-1}$ and with nutrient solution at EC $1.2dS{\cdot}m^{-1}$. The air was periodically circulated between the two chambers using a diaphragm pump. The $CO_2$ emission rate of the mushroom increased until the $15^{th}$ day after scratching (DAS) and then decreased. The rate also increased with increased indoor temperature. In particular, the $CO_2$ emission rate per fresh weight of fruit body increased by about 3.1 times after thinning compared to before thinning. In terms of $CO_2$ balance, the $CO_2$ emission rates from a bottle (950 mL) of the mushroom at 9, 12, and 14 DAS were equivalent to those of 3, 4.5, and 5.5 lettuce plants at 7, 10, and 12 DAT (days after transplanting), respectively. This work shows that balance in $CO_2$ concentration could be achieved using an appropriate ratio of the two crops in a closed production system.

Effect of Temperature on Development and Reproduction of the Cotton Caterpillar, Palpita indica(Lepidoptera: Pyralidae) (목화바둑명나방(나비목:명나방과)의 발육과 생식에 미치는 온도의 영향)

  • Shin, Wook-Kyun;Kim, Gil-Hah;Song, Cheol;Kim, Jeong-Wha;Cho, Kwang-Yun
    • Korean journal of applied entomology
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    • v.39 no.3
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    • pp.135-140
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    • 2000
  • Development and reproduction of the cotton caterpillar, Palpita indica, were investigatedunder different temperatures (15 .O, 17.5, 20.0, 22.5, 25 .O, 27.5, 30.0, 32.5, and 35 .O$^{\circ}$C). Duration fromegg to pre-adult of the cotton caterpillar were ranged from 68.6 days at 175$^{\circ}$C to 19.7 days at 35.0% (3.5times shorter growth period compared with that at 17S$^{\circ}$C). At 15.0$^{\circ}$C, cotton caterpillar eggs developedto the last larval instar but were not able to go through the pupal stage. The lower developmentalthreshold temperatures and degree-days of egg, larva, pupa, and complete development were 13.4, 10.6,11.6, and 11.5"C and 55.3,251.5, 138.3, and 479.8 degree days, respectively. The hatching, pupation andemergence rates were higher at 25.0eC and 27.5"C compared with other temperatures. The survival ratefrom the hatched larva to adult was the highest at 27.5"C. The preoviposition and the adult longevity were11.5 and 30.6 days at 17.5"C and 1.5 and 9.2 days at 35.0$^{\circ}$C, respectively. The mean fecundity perfemales was greater at 25.0$^{\circ}$C and 27.5"C compared with other temperatures. Mean generation time indays (T) was shorter on higher temperature. Net reproductive rate per generation (R,) was the lowest atthe highest temperature as well as at the lowest, and it was 199.1 which was the highest at 27.5"C. Theintrinsic rate of natural increase (r,) was highest at 30.0$^{\circ}$C as 0.148. As a result, optimum ranges oftemperature for P. indica growth were between 25.0-32.5"C .emperature for P. indica growth were between 25.0-32.5"C .t;C .

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Human Blastocysts;The Correlation Between Embryo Microscopical Assessments and Their Cell Number (인간 배반포기 배의 현미경적 분류와 세포수의 상관관계에 관한 연구)

  • Kim, E.Y.;Uhm, S.J.;Kim, M.K.;Yoon, S.H.;Park, S.P.;Chung, K.S.;Lim, J.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.23 no.3
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    • pp.319-326
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    • 1996
  • The objective of this study was to investigate correlation between the morphology by microscopic assessments of surplus blastocysts produced in human IVF program and their cell number obtained by differential labelling method. For these experiments, 76 surplus human blastocysts were obtained from 36 patients on day 5 after IVF, the embryos were classified to early (ErB), early expanding (EEB), middle expanding (MEB), expanded blastocyst (EdB) according to their blastocoel expansion and zona thickness. When the ovum size and zona thickness of the classified blastocysts were measured using micrometer, although the embryos were produced in the same culture condition, there were significant variances in ovum size ($148.8 217.6{\mu}m$) and zona thickness ($1.2-14.4{\mu}m$). Total blastomere cell number counted after hoechst staining was increased by two to three fold during the transition period from ErB ($39.1{\pm}3.6$) to EdB ($(89.6{\pm}3.3)$) stage on day 5 after IVF. ICM ($11.9{\pm}1.8-22.2{\pm}4.3$) and TE ($24.5{\pm}3.6-70.0{\pm}7.7$) cell numbers using differential labelling were also showed the increased pattern according to the developmental level. Especially, EdB which showed poor ICM morphologically also indicated the low ICM cell number after differential labelling. This demonstrated that there is good correlation between the morphological assessment and the cell number. The count of ICM and TE nuclei using differential labelling can be used as an important criterion, if it is accompanied with morphological assessments, in selecting the better embryos for improving the pregnancy rates in human blastocyst transfer program.

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Studies on the Bionomic of Gonatocerus sp. y (Hymenoptera: Mymaridae), an Egg Parasite of the Green Rice Leafhopper, Nephotettix cincticeps Uhler (끝동매미충 난기생봉(Gonatocerus sp. y)의 생태에 관한 연구)

  • Kim Jeoung Bu;Kim Chang Hyo
    • Korean journal of applied entomology
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    • v.23 no.3 s.60
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    • pp.158-165
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    • 1984
  • These studies were carried out to investigate the bionomics of Gonatocerus sp. y (Hymenoptera: Mymaridae) , an egg parasite of the green rice leafhopper, Nephotettix cincticeps Uhler (Homoptera: Deltocephalidae) under laboratory and field conditions at Gyeongnam Provincial O.R.D. in Jinju from 1982 to 1983. The developmental period of each life stage was observed under four different temperature conditions from $15^{\circ}C\;to\;30^{\circ}C\;with\;5^{\circ}C$ increment. The development from egg to adult at 15^{\circ}C,\;20^{\circ}C,\;25^{\circ}C,\;and\;30^{\circ}C $ was completed in an average of 53, 25, 15, and 12 days and the females laid an average of 28, 48, 42, and 32 eggs respectively, with a maximum of 98. And most of the eggs were laid in the first two days after emergence. The time of adult emergence was determined under $25{\pm}1^{\circ}C$ condition. More than $76\%$ of the wasps emerged in the morning and the rest of them emerged in the afternoon. The average sex ratio between male and female was 1 : 5 when the wasps were developed from fertilized eggs. The longevity of female was greater than that of male when honey and water were provided as food, however, it was not when only water was supplied as food. The average longevities of females were 30 days at $15^{\circ}C$ and 15 days at $20^{\circ}C$ when honey and water were given as food. The peak precent parasitism of Gonatocerus sp. v on Nephotettix cincticeps in fallow fields was observed between late April and early May. The percent parasitism of the wasps In fallow fields was an average of 49\~59.9\% $ in Gyeochang, Namji and $23.2\~36\%$ in Namhae. The parasitic activities of the parasite had four peaks in paddy field, however, the highest peak was observed between late August and early September. The Percent e99 Parasitism of Gonatocerus sp. y was an average of $12.3\%$ on second the generation and $24.6\%$ of third genera lion of Nephotettix cinrticeps.

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Monocentric Derivative Y Chromosome with Duplication of the SRY Gene in an Azoospermic Male (무정자증 불임남성에서 관찰된 SRY 유전자의 중복을 동반한 일동원체성 derivative Y 염색체)

  • Choi, Eun-Young;Lee, Bom-Yi;Park, Ju-Yeon;Lee, Yeon-Woo;Oh, Ah-Rum;Lee, Shin-Young;Kim, Shin-Young;Han, You-Jung;Lee, Mee-Bum;Ryu, Hyun-Mee;Seo, Ju-Tae;Park, So-Yeon
    • Journal of Genetic Medicine
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    • v.7 no.2
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    • pp.160-164
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    • 2010
  • Structural abnormalities of the Y chromosome affect normal testicular differentiation and spermatogenesis. The present case showed a rare monocentric derivative Y chromosome with partial duplication of Yp including the SRY gene and deletion of Yq12 heterochromatin. The karyotype was 46,X,der(Y)(pter${\rightarrow}$q11.23::p11.2${\rightarrow}$pter).ish der(Y)(DYZ3+,DYZ1-,SRY++), confirmed through a FISH study. Even though the patient possessed an abnormal Y chromosome, testicular biopsy showed normal testicular volumes in the proband, with gonadal hormonal levels in the normal range but bilateral varicocele and hypospermatogenesis. We speculate that the abnormal Y chromosome arose from sister chromatids during Y chromosome recombination or intra chromosomal NAHR (non-allelic homologous recombination) during meiosis in the patient's father or in the very early stages of embryogenesis. The derivative Y chromosome might interfere in the meiotic stage of spermatogenesis, leading to the developmental arrest of germ cells. The present case illustrates that the infertility phenotype can have various causes. Also, it emphasizes the importance of accurate and various genetic analyses and could aid in male infertility treatment.

Studies on Ecological Environments and Indoor-rearing Conditions of the Common Grass Yellow Butterfly, Eurema hecabe (남방노랑나비(Eurema hecabe)의 생태환경 및 실내사육 조건에 관한 연구)

  • Lee, Sang-Hyun;Kim, Se-Gwon;Nam, Gyoung-Pil;Son, Jai-Duk;Lee, Jin-Gu;Park, Young-Kyu;Choi, Young-Cheol;Lee, Young-Bo
    • Journal of Sericultural and Entomological Science
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    • v.50 no.2
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    • pp.133-139
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    • 2012
  • The Aims of this study are to investigate ecological environments and indoor-rearing conditions of Eurema hecabe for artificial rearing. Two different nesting site, a open area nearby small river(A area; Daejin-ri, Gonyang-myeon, Sachun-si, Gyoungsangnam-do) and a foot of a hill area(B area; Noryang-ri, Seolcheon-myeon, Namhae-gun, Kyoungsangnam-do), selected for investigation of ecological habitat environment, and investigated two times per month, during six month, from June to November in 2010. In result, the number of butterflies, Eurema hecabe, at the hill area(B area) was 6 times more than the open area(A area). We observed the number of eggs, larvae and puapae in the B area more times compared to those in the A area. In the B area, Lespedeza cuneata was higher preference than the other host plant, Lespedeza pilosa and Albizia julibrissin. For development of indoor-rearing conditions, common grass yellow butterfly, Eurema hecabe, was reared in a room condition. As a result of oviposition preference experiment, 100-mated-female laid significantly higher number of eggs $104.9{\pm}19.6$ on Lespedeza cuneata plant, whereas the number of eggs was $12.7{\pm}4.5$ on Cassia obtusifolia plant. Moreover, the eggs hatched within $5.1{\pm}0.9$ days from the day of oviposition and procent of egg hatchability was 53.7% on host plant. The headwidth of each developmental larval stage were $0.36{\pm}0.02$(1st), $0.61{\pm}0.02$(2nd), $0.93{\pm}0.05$(3rd), $1.46{\pm}0.08$(4th), $2.25{\pm}0.11$(5th). The larval period was $12.1{\pm}0.9$ days under high temperature, long day condition($25^{\circ}C$, 16L : 8D), showing 81.0% pupal ratio. The pupal period was $6.9{\pm}0.7$ days, and the emergence rate was 79.6%. Based on above experiment, artificial rearing system of common grass yellow butterfly, Eurema hecabe was completed in indoor condition.

A Study on Classification System for Gong-Po-Do Style in Tomb Wall Paintings of Koguryo (고구려 고분벽화 공포도 형식의 분류체계에 관한 연구)

  • Hwang, Se-Ok
    • Korean Journal of Heritage: History & Science
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    • v.49 no.2
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    • pp.20-55
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    • 2016
  • Koguryo's tomb mural paintings in North Korea are our precious cultural heritage which have been designated as UNESCO World Heritage property receiving high praise in the following criterion, i) exceptional creativeness of human being, ii) representative value showing the stage of development in construction history of East-Asia, iii) aesthetic superiority iv) uniqueness of building construction including tombs' ceiling. Mural paintings have been found from almost 100 tombs of the Koguryo dynasty out of 130 which are scattered across Huanren County, Lianoning Province, Ji'an, Jilin Province in China and Pyongyang in North Korea. Especially, most of them are gathered in Pyongyang from 4th and 5th century. Peculiarly, some of them have been constructed before King Jangsu's transfer of the capital to Pyongyang(AD 427). It can be regarded that Pyongyang territory had been under control of Koguryo and to become a new capital in the near future. And dense emergence of such tombs since the capital transfer from Gungnae City to Pyongyang during the reign of Jangsu is linked closely to the construction of tombs for rulers under strengthen royal authority of Jangsu and centralized system of authoritarian rule. Tomb mural paintings describe the owner's figure pictorially based on the truth just as in his living years. General lifestyles of ruling powers and sovereigns can be seen from the wall paintings portraying several buildings with various styles, figures, manners of living, which are considered that the tomb owner had led politically and sociologically in his life. In spite of not enough proofs to approve figure of architectures or "Gong-Po" in wall paintings on the tombs as those of Koguryo, it is persuasive with consideration for painting and decoration inside the tomb like wooden building in real life for the purpose of reenacting and continuing the tomb owner's luxurious life after death. "Du-Gong-Po-Zak" had appeared in company with Koguryo tomb murals and it can be found in most of the murals. And the emergence of substantial "Gong-Po-Do" can be counted more than a century ahead of the figure in murals. It could be a reasonable assumption as regards Koguryo tomb murals time of appearance match up with production period of Gahyungmyunggi(家形明器) and Hwasangseok(畵像石) Hwasangjeon(畵像塼) Design in the Mural Painting of the East-Han(東漢) Ancient Tombs in China. On this study, architectural "Gong-Po"s described in Koguryo tomb murals are categorized largely in "Bi(non)-Po-Zak-kye", "Jun(semi)-Po-Zak-kye", and "Po-Zak-kye" based on presence of "Ju-Du", "Cheom-Cha", and "Cheom-Cha-Sal-Mi" with developmental aspect, and, "Po-zak" is subdivided as "Bi(non)-Cheul-Mok" and "Cheul-Mok" types.