• Title/Summary/Keyword: microscopic analysis

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Improving Corsican pine somatic embryo maturation: comparison of somatic and zygotic embryo morphology and germination

  • Wtpsk, Senarath;Shaw, D.S.;Lee, Kui-Jae;Lee, Wang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.04a
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    • pp.61-62
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    • 2003
  • Clonal propagation of high-value forest trees through somatic embryogenesis (SE) has the potential to rapidly capture the benefits of breeding or genetic engineering programs and to improve raw material uniformity and quality. A major barrier to the commercialization of this technology is the low quality of the resulting embryos. Several factors limit commercialization of SE for Corsican pine, including low initiation rates, low culture survival, culture decline causing low or no embryo production, and inability of somatic embryos to fully mature, resulting in low germination and reduced vigour of somatic seedlings. The objective was to develop a Corsican pine maturation medium that would produce cotyledonary embryos capable of germination. Treatments were arranged in a completely randomized design. Data were analyzed by analysis of variance, and significant differences between treatments determined by multiple range test at P=0.05. Corsican pine (Pinus nigra var. maritima) cultures were initiated on modified !P6 medium. Modifications of the same media were used for culture multiplication and maintenance. Embryogenic cultures were maintained on the same medium semi solidified with 2.5 g/l Gelrite. A maturation medium, capable of promoting the development of Corsican pine somatic embryos that can germinate, is a combination of iP6 modified salts, 2% maltose, 13% polyethylene glycol (PEG), 5 mg!l abscisic acid (ABA), and 2.5 g/l Gelrite. After initiation and once enough tissue developed they were grown in liquid medium. Embryogenic cell suspensions were established by adding 0.951.05 g of 10- to 14-day-old semisolid-grown embryogenic tissue to 9 ml of liquid maintenance media in a 250ml Erlenmeyer flask. Cultures were then incubated in the dark at 2022$^{\circ}$C and rotated at 120 rpm. After 2.53 months on maturation medium, somatic embryos were selected that exhibited normal embryo shape. Ten embryos were placed horizontally on 20 ml of either germination medium ($\frac{2}{1}$strength Murashige and Skoog (1962) salts with 2.5 g/l activated charcoal) or same medium with copper sulphate adjusted to 0.25 mg/1 to compensate for copper adsorption by activated carbon. 2% and 4% maltose was substituted by 7.5% and 13% PEG respectively to improve the yield of the embryos. Substitution of' maltose with PEG was clearly beneficial to embryo development. When 2% of the maltose was replaced with 7.5% PEG, many embryos developed to large bullet-shaped embryos. At latter stages of development most embryos callused and stopped development. A few short, barrel-shaped cotyledonary embryos formed that were covered by callus on the sides and base. When 4% of the maltose was removed and substituted with 13% PEG, the embryos developed further, emerging from the callus and increasing yield slightly. Microscopic examination of the cultures showed differing morphologies, varying from mostly single cells or clumps to well-formed somatic embryos that resembled early zygotic embryos only liquid cultures with organized early-stag. A procedure for converting and acclimating germinants to growth in soil and greenhouse conditions is also tested. Seedling conversion and growth were highly related to the quality of the germinant at the time of planting. Germinants with larger shoots, longer, straighter hypocotyls and longer roots performed best. When mature zygotic embryos germinate the root emerges, before or coincident with the shoot. In contrast, somatic embryos germinate in reverse sequence, with the cotyledons greening first, then shoot emergence and then, much later, if at all, the appearance of the root. Somatic seedlings, produced from the maturation medium, showed 100% survival when planted in a field setting. Somatic seedlings showed normal yearly growth relative to standard seedlings from natural seed.

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High Glucose Induces Apoptosis through Caspase-3 Dependent Pathway in Human Retinal Endothelial Cell Line (인간망막 내피세포주에서 고농도 포도당이 caspase-3 경로를 통해 세포자연사 유도)

  • Seo, Eun-Sun;Chae, Soo-Chul;Kho, Eun-Gyeong;Lee, Jong-Bin
    • Korean Journal of Environmental Biology
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    • v.27 no.1
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    • pp.66-72
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    • 2009
  • Diabetic Retinopathy (DR) is a leading cause of blindness among adults in the western countries. Hyperglycemia is a condition, that induces apoptotic cell death in a variety of cell types in diabetes, but the mechanism remains unclear. The aim of the study is to understand the effects of high Glucose on Human Retinal Endothelial Cells. Retinal endothelial cells were cultured in Iscove's Modified Dulbecco's Medium (IMDM) containing 5, 25 and 50 mM Glucose, incubated for 24, 36 and 48 hours in humidified 5 % CO$_2$ incubator at 37$^{\circ}C$. Human Retinal Endothelial Cell Line (HREC) were characterized for morphology with different treatment by phase contrast microscopic analysis. Number of dead and viable cells was counted by trypan blue exclusion and supported by MTT assay. The intracellular Hydrogen peroxide (H$_2$O$_2$), a Reactive Oxygen Species (ROS) generation in high glucose conditions was assessed by FOX II assay and apoptosis by caspase-3 assay. The high glucose treated cells undergoing DNA fragmentation was witnessed by Agarose gel electrophoresis. We found that the cells incubated with 25 and 50 mM glucose containing medium for 48 hours altered the morphology of the cell, induced apoptosis and DNA fragmentation. The dead cell number were high in 25 and 50 mM when compared to the cells incubated with 5 mM glucose for 24, 36, and 48 hours. Also, the H$_2$O$_2$ levels and the activity of caspase-3 were increased in high glucose treated cells. Conclusions/interpretation: Our results demonstrated that elevated glucose induces apoptosis in cultured HREC. The hyperglycemia-induced increase in apoptosis may be dependent on caspase activation. The association between ROS generation and caspase-3 activation on high glucose treated cells is yet to be investigated.

Histological Analysis of Autologous Pericardial Tissue Used as a Small-Diameter Arterial Graft (소구경 동맥이식편으로 사용한 자가심란의 조직학적 분식)

  • Yang Ji-Hyuk;Sung Sang-Hyun;Kim Won-Gon
    • Journal of Chest Surgery
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    • v.39 no.4 s.261
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    • pp.261-268
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    • 2006
  • Background: Current vascular prostheses are still inadequate for reconstruction of small-diameter vessels. Autologous pericardium can be a good alternative for this purpose as it already possesses good blood compatibility and shows a mechanical behavior similar to that of natural arteries. However, the clinical use of autologous pericardial tissue as a small-diameter vascular graft has limitations due to mixed outcomes from uncertain biological behavior and difficulty to gain reliable patency results in animal experiments. To study this issue, we implanted fresh and glutaraldehyde-treated autologous pericardium as small-diameter arterial grafts in dogs, and compared their time-related changes histologically. Material and Method: As a form of 5mm-diameter arterial graft, one pair of autologous pericardial tissue was used for comparison between the glutaraldehyde-treated and the glutaraldehyde-untreated grafts in the bilateral carotid arteries in the same dog. The patency of the grafts were evaluated at regular intervals with Doppler ultrasonography. After the predetermined periods of 3 days, 2 weeks, 1 month, 3 months and 6 months, the grafts in each animal were explanted. The retrieved grafts were processed for light and electron microscopic analyses following gross observation. Result: Of 7 animals, 2 were excluded from the study because one died postoperatively due to bleeding and the other was documented as one side of the grafts being obstructed. All 10 grafts in the remaining 5 dogs were patent. Grossly, a variable degree of thromboses were observed in the luminal surfaces of the grafts at 3 days and 2 weeks, despite good patency. Pseudointimal smooth blood-contacting surfaces were developed in the grafts at f month and later. By light microscopy, mesothelial cell layers of the pericardial tissue were absent in all explanted grafts. Newly formed endothelial cell layers on the blood-contacting surface were observed in both the glutaraldehyde-treated and fresh grafts at 3 months and later. The collagen fibers became degraded by fragmentation in the fresh graft at 1 month and In the glutaraldehyde-treated graft at 3 months. At 6 months, the collagen layers were no longer visible in either the glutaraldehyde-treated or fresh grafts. By electron microscopy, a greater amount of coarse fibrin fibers were observed in the fresh grafts than in the glutaraldehyde-treated grafts and, more compact and well-arrayed layers were observed in the glutaraldehyde-treated grafts than in the fresh grafts. Conclusion: The glutaraldehyde-treated small-diameter pericardial arterial grafts showed a better endothelialization of the blood-contacting surface and a slower fragmentation of the collagen layers than the fresh grafts, although it has yet to be proven whether these differences are so significant as to affect the patency results between the groups.

Studies on the Structure and Some Physical and Chemical Properties of the Egg Shell in the Silkworm, Bombyx mori L. (가잠난각의 구조 및 물리화학적 특성에 관한 연구)

  • 마영일;박광의
    • Journal of Sericultural and Entomological Science
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    • v.24 no.2
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    • pp.55-72
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    • 1983
  • These studies were done to find out any difference, ultrastructural, physical or chemical, between the shells of diapausing and non-diapausing eggs of the silkworm, Bombyx mori L. 1. From the electron-microscopic observation, the egg shells have four distinctive layers. In addition to the four layers, the shells in the diapausing eggs has another layer with low electron density on its surface. 2. The permeability of the egg shell to hydrochloride was much lower in diapausing egg than in non-diapausing egg. Also the permeability changed in the opposite directions with the egg age: the diapausing eggs decreased while non-diapausing ones increased. 3. The permeability increased when the diapausing egg shell was treated with HCl. When they were treated with ether, however, the increase in permeability was much smaller. It seems there was an ether soluble material involved in the content of the egg shell. 4. The diapausing eggs were also much more resistant to desiccation than the non-diapausing ones. The former, when treated with HCl or chilling, became less resistant to desiccation. 5. The positive histochemical response of the egg shell to PAS-Alcian blue and protein stainings suggests presence of abundant proteins and carbohydrates in the egg shell. On the other hand, the staining response to lipid was more positive in the inner layers than in the outer layer of the shell. 6. The egg shell adhesives seems to be mucopolysaccharides produced by colleterial glands, since the oviposited eggs showed a positive responses to carbohydrate and negative to lipid-staining chemicals, but not the mature oocytes in the ovarioles. 7. There were two bands on the electrophoretic pattern of the SH proteins extracted from the egg shells both in the diapausing egg and non-diapausing one: a slow moving major component and a fast moving minor one. However, the electrophoretic mobility showed a difference in the minor components between them. It is evident that the fast moving minor one of non-diapausing egg ran a little further than that of diapausing egg. 8. In amino acids analysis, no significant differences were found in their composition between diapausing and non-diapausing egg and SH proteins contain relatively more glycine and less cystine.

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Photodynamic Therapy with Photofrin Reduces Invasiveness of U87 Malignant Human Glioma Cells (교모세포종 세포주 U87에서 Photofrin을 사용한 광역학 치료가 종양 침습성에 미치는 영향)

  • Woo, Hye Kyung;Cho, Kyung-Keun;Rha, Hyung Kyun;Lee, Kyung Jin;Park, Sung Chan;Cho, Jung Ki;Park, Hea Kwan;Kang, Joon Ki;Choi, Chang Rak
    • Journal of Korean Neurosurgical Society
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    • v.30 no.sup2
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    • pp.189-196
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    • 2001
  • Objective : We tested the hypothesis that photodynamic therapy(PDT) with Photofrin inhibits tumor invasion of U87 human glioma cells using several in vitro assay to measure tumor invasiveness. The effects of PDT on cell growth, directional migration and cell invasion were investigated. Material and Method : Tumor cells were treated with Photofrin at various doses and at a fixed optical(632nm) dose of $100mJ/cm^2$. Cytotoxicity was tested using the MTT method. Invasion assays including the matrigelartificial basement membrane barrier migration and spheroid confrontation with confocal microscopic analysis were used to study the relationship between PDT and invasiveness. Result : U87 cells showed a dose dependent cytotoxic response to increasing Photofrin dose. Data from the matrigel artificial basement membrane assay indicate that PDT inhibits the U87 cell migration dose dependently. Low doses of subcytotoxic PDT treatment, such as 2.5ug/ml Photofrin dose, also appeared to significantly inhibit migration of U87 cells(p<0.05). In co-cultures between U87 cell spheroids and brain aggregates, progressive invasion with destruction of the brain aggregate occurs. The extent of tumor cell infiltration and proportion or intact brain aggregate remaining after 24h differs in Photofrin PDT treated versus Photofrin only control, with changes suggestive of a dose-response effect. Conclusion : our data indicate that PDT with Photofrin significantly inhibits the invasiveness of U87 cells, and this inhibition is dose dependent.

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EVALUATION OF PERIODONTAL LIGAMENT CELL VIABILITY IN RAT TEETH AFTER FROZEN PRESERVATION USING IN-VIVO MTT ASSAY (급속냉동된 쥐 치아의 in vivo MTT 검색법을 이용한 치주인대세포 활성도 평가)

  • Kim, Jae-Wook;Kim, Eui-Sung;Kim, Jin;Lee, Seung-Jong
    • Restorative Dentistry and Endodontics
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    • v.31 no.3
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    • pp.192-202
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    • 2006
  • The purpose of this study was to examine the viability of PDL cells in rat molars by using in vivo MTT assay, which was used to compare fast cryopreservation group by liquid nitrogen $(-196^{\circ}C)\;with\;4^{\circ}C$ cold preservation group. A total of 74 Sprague-Dawley white female rats of 4 week-old with a body weight of 100 grams were used. The maxillary left and right, first and second molars were extracted as atraumatically as possible under ketamine anesthesia. Ten teeth of each group were divided as six experimental groups depending upon the preservation. Cryopreservation groups were Group 1 (5% DMSO 6% HES in F medium) Group 2 (10% DMSO in F medium), Group 3 (5% DMSO 6% HES in $Viaspan^(R)$). Group 4 (10% DMSO in $Viaspan^(R)$) which were cryopreserved for 1 week and cold preservation groups were Group 5 (F medium) , Group 6 ($Viaspan^(R)$) at $4^{\circ}C$ for 1 week. Immediate extraction group was used as a control. After preservation and thawing, the in vivo MTT assay was processed. Two way ANOVA and Duncan's Multiple Range Test was performed at the 95 % level of confidence, Another 2 teeth of each group were treated as the same manner and frozen sections $10{\mu}m$ thick for microscopic observation. The value of optical density obtained after in vivo MTT analysis was divided by the value of eosin staining for tissue volume standardization. Group 1, 2 had significantly higher optical density than Group 3 and 4 which had the lowest OD value. Group 6 had higher OD value than in Group 5 (P<0.05). Histological findings of periodontal ligament cell, after being stained with MTT solution were consistent with the in vivo MTT assay results. In this study, the groups which were frozen with DMSO as a cryoprotectant and the groups with F medium showed the best results.

Pseudotachylyte Developed in Granitic Gneiss around the Bulil Waterfall in the Jirisan, SE Korea: Its Occurrence and Characteristics (지리산 불일폭포 일원의 화강암질편마암에 발달한 슈도타킬라이트: 산상과 특성)

  • Kang, Hee-Cheol;Kim, Chang-Min;Han, Raehee;Ryoo, Chung-Ryul;Son, Moon;Lee, Sang-Won
    • The Journal of the Petrological Society of Korea
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    • v.28 no.3
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    • pp.157-169
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    • 2019
  • Pseudotachylytes, produced by frictional heating during seismic slip, provide information that is critical to understanding the physics of earthquakes. We report the results of occurrence, structural characteristics, scanning electron microscopic observation and geochemical analysis of pseudotachylytes, which is presumed to have formed after the Late Cretaceous in outcrops of the Paleoproterozoic granitic gneiss on the Bulil waterfall of the Jirisan area, Yeongnam massif, Korea. Fault rocks, which are the products of brittle deformation under the same shear stress regime in the study area, are classified as pseudotachylyte and foliated cataclasite. The occurrences of pseudotachylyte identified on the basis of thickness and morphology are fault vein-type and injection vein-type pseudotachylyte. A number of fault vein-type pseudotachylytes occur as thin (as thick as 2 cm) layers generated on the fault plane, and are cutting general foliation and sheared foliation developed in granitic gneiss. Smaller injection vein-type pseudotachylytes are found along the fault vein-type pseudotachylytes, and appear in a variety of shapes based on field occurrence and vein geometry. At a first glance fault vein-type seudotachylyte looks like a mafic vein, but it has a chemical composition almost identical to the wall rock of granitic gneiss. Also, it has many subrounded clasts which consist predominantly of quartz, feldspar, biotite and secondary minerals including clay minerals, calcite and glassy materials. Embayed clasts, phenocryst with reaction rim, oxide droplets, amygdules, and flow structures are also observed. All of these evidences indicate the pseudotachylyte formed due to frictional melting of the wall rock minerals during fault slip related to strong seismic faulting events in the shallow depth of low temperature-low pressure. Further studies will be conducted to determine the age and mechanical aspect of the pseudotachylyte formation.

An Analysis of the Uses of External Representations in Matter Units of 7th-Grade Science Digital Textbooks Developed Under the 2015 Revised National Curriculum (2015 개정 교육과정에 따른 중학교 1학년 디지털교과서의 물질 단원에서 나타난 외적 표상의 활용 실태 분석)

  • Song, Nayoon;Hong, Juyeon;Noh, Taehee
    • Journal of the Korean Chemical Society
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    • v.64 no.6
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    • pp.416-428
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    • 2020
  • This study analyzed the uses of external representations presented in the matter units of the 7th-grade science digital textbooks developed under the 2015 revised national curriculum. The level, form, presentation, and interactivity of external representations presented in 5 types of digital textbooks were analyzed. As for the level, the macroscopic level of representations was mainly presented. The macroscopic level and microscopic level of representations were presented together in the particle description. As for the form, visual-verbal and visual-nonverbal representations were usually presented across the board. Very few audial-verbal and audial-nonverbal representations were presented. Visual-verbal and audial-verbal representations were mostly presented in formal form, and visual-nonverbal representations were mostly presented in illustration without movement. The presentation of representations was analyzed in three aspects. First, visual-verbal and visual-nonverbal representations were mainly presented together and none of audial-verbal and visual-nonverbal representations were presented together. When the representations of the audial-verbal, visual-nonverbal, and visual-verbal were presented together, some of the information presented in audial-verbal representations was repeatedly presented in the visual-verbal representations. Second, audial-nonverbal representations not related to learning content were presented along with other representations. Third, there were few cases of arranging visual-verbal and visual-nonverbal representations on the next pages. Audialverbal and visual-nonverbal representations were always presented synchronized. As for the interactivity, the manipulation level was mainly presented in the main area, and the feedback level was mainly presented in the activity area. The adaptation level and the communication level of interactivity were presented very few. Based on the results, the implications for the direction of constructing digital textbooks were discussed.

Raman Spectroscopic Study for Investigating the Spatial Distribution and Structural Characteristics of Mn-bearing Minerals in Non-spherical Ferromanganese Nodule from the Shallow Arctic Ocean (북극해 천해저 비구형 망가니즈단괴 내 광물종 분포 및 구조적 특성 규명을 위한 라만 분광분석 연구)

  • Sangmi, Lee;Hyo-Jin, Koo;Hyen-Goo, Cho; Hyo-Im, Kim
    • Korean Journal of Mineralogy and Petrology
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    • v.35 no.4
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    • pp.409-421
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    • 2022
  • Achieving a highly resolved spatial distribution of Mn-bearing minerals and elements in the natural ferromanganese nodules can provide detailed knowledge of the temporal variations of geochemical conditions affecting the formation processes of nodules. While a recent study utilizing Raman spectroscopy has reported the changes in the manganate mineral phases with growth for spherical nodules from the Arctic Sea, the distributions of minerals and elements in the nodules from the shallow Arctic Sea with non-spherical forms have not yet fully elucidated. Here, we reported the micro-laser Raman spectra with varying data acquisition points along three different profiles from the center to the outermost rim of the non-spherical ferromanganese nodules collected from the East Siberian Sea (~73 m). The elemental distributions in the nodule (such as Mn, Fe, etc.) were also investigated by energy dispersive X-ray spectroscopy (EDS) analysis to observe the internal structure and mineralogical details. Based on the microscopic observation, the internal structures of a non-spherical nodule can be divided into three different regions, which are sediment-rich core, iron-rich substrate, and Mn-Fe layers. The Raman results show that the Mn-bearing mineral phases vary with the data acquisition points in the Mn-Fe layer, suggesting the changes in the geochemical conditions during nodule formation. In addition, we also observe that the mineral composition and structural characteristics depend on the profile direction from the core to the rim. Particularly, the Raman spectra obtained along one profile show the lack of Fe-(oxy)hydroxides and the noticeably high crystallinity of Mn-bearing minerals such as birnessite and todorokite. On the other hand, the spectra obtained along the other two profiles present the presence of significant amount of amorphous or poorly-ordered Fe-bearing minerals and the low crystallinity of Mn-bearing minerals. These results suggest that the diagenetic conditions varied with the different growth directions. We also observed the presence of halite in several layers in the nodule, which can be evidence of the alteration of seawater after nodule formation. The current results can provide the opportunity to obtain detailed knowledge of the formation process and geochemical environments recorded in the natural non-spherical ferromanganese nodule.

A Study on the Conservation State and Plans for Stone Cultural Properties in the Unjusa Temple, Korea (운주사 석조문화재의 보존상태와 보존방안에 대한 연구)

  • Sa-Duk, Kim;Chan-Hee, Lee;Seok-Won, Choi;Eun-Jeong, Shin
    • Korean Journal of Heritage: History & Science
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    • v.37
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    • pp.285-307
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    • 2004
  • Synthesize and examine petrological characteristic and geochemical characteristic by weathering formation of rock and progress of weathering laying stress on stone cultural properties of Unjusa temple of Chonnam Hwasun county site in this research. Examine closely weathering element that influence mechanical, chemical, mineralogical and physical weathering of rocks that accomplish stone cultural properties and these do quantification, wish to utilize by a basic knowledge for conservation scientific research of stone cultural properties by these result. Enforced component analysis of rock and mineralogical survey about 18 samples (pyroclastic tuff; 7, ash tuff; 4, granite ; 4, granitic gneiss; 3) all to search petrological characteristic and geochemical characteristic by weathering of Unjusa temple precinct stone cultural properties and recorded deterioration degree about each stone cultural properties observing naked eye. Major rock that constitution Unjusa temple one great geological features has strike of N30-40W and dip of 10-20NE being pyroclastic tuff. This pyroclastic tuff is ranging very extensively laying center on Unjusa temple and stone cultural properties of precinct is modeled by this pyroclastic tuff. Stone cultural propertieses of present Unjusa temple precinct are accomplishing structural imbalance with serious crack, and because weathering of rock with serious biological pollution is gone fairly, rubble break away and weathering and deterioration phenomenon such as fall off of a particle of mineral are appearing extremely. Also, a piece of iron and cement mortar of stone cultural properties everywhere are forming precipitate of reddish brown and light gray being oxidized. About these stone cultural properties, most stone cultural propertieses show SD(severe damage) to MD(moderate damage) as result that record Deterioration degree. X-ray diffraction analysis result samples of each rock are consisted of mineral of quartz, orthoclase,plagioclase, calcite, magnetite etc. Quartz and feldspar alterated extremely in a microscopic analysis, and biotite that show crystalline form of anhedral shows state that become chloritization that is secondary weathering mineral being weathered. Also, see that show iron precipitate of reddish brown to crack zone of tuff everywhere preview rock that weathering is gone deep. Tuffs that accomplish stone cultural properties of study area is illustrated to field of Subalkaline and Peraluminous, $SiO_2$(wt.%) extent of samples pyroclastic tuff 70.08-73.69, ash tuff extent of 70.26-78.42 show. In calculate Chemical Index of Alteration(CIA) and Weathering Potential Index(WPI) about major elements extent of CIA pyroclastic tuff 55.05-60.75, ash tuff 52.10-58.70, granite 49.49-51.06 granitic gneiss shows value of 53.25-67.14 and these have high value gneiss and tuffs. WPI previews that is see as thing which is illustrated being approximated in 0 lines and 0 lines low samples of tuffs and gneiss is receiving esaily weathering process as appear in CIA. As clay mineral of smectite, zeolite that is secondary weathering produce of rock as result that pick powdering of rock and clothing material of stone cultural properties observed by scanning electron micrographs (SEM). And roots of lichen and spore of hyphae that is weathering element are observed together. This rock deep organism being coating to add mechanical weathering process of stone cultural properties do, and is assumed that change the clay mineral is gone fairly in stone cultural properties with these. As the weathering of rocks is under a serious condition, the damage by the natural environment such as rain, wind, trees and the ground is accelerated. As a counter-measure, the first necessary thing is to build the ground environment about protecting water invasion by making the drainage and checking the surrounding environment. The second thing are building hardening and extirpation process that strengthens the rock, dealing biologically by reducing lichens, and sticking crevice part restoration using synthetic resin. Moreover, it is assumed to be desirable to build the protection facility that can block wind, sunlight, and rain which are the cause of the weathering, and that goes well with the surrounding environment.