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Inhibitory Effects of Ethanol Extracts from Nuruk on Oxidative Stress, Melanogenesis, and Photo-Aging

  • Lee, Sang-Jin;Cho, Sung-Won;Kwon, Yi-Young;Kwon, Hee-Suk;Shin, Woo-Chang
    • Mycobiology
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    • v.40 no.2
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    • pp.117-123
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    • 2012
  • Nuruk contributes to the unique characteristics of Korean alcoholic beverages. In this study, the effects of nuruk extracts (NE) on anti-oxidant characters, melanogenesis, and anti-photoaging activity were investigated. NEs were obtained from the 70% ethanol extracts of six types of nuruk, which have been used in brewing of fermented alcohol beverages in Korea. First, various antioxidant characteristics were identified in terms of 2,2'-azino-bis(3-ethylbenzthiozoline-6-sulphonic acid) (ABTS) radical scavenging activity, superoxide dismutase (SOD) expression, and inhibition of xanthine oxidase activity. NE#4 exhibited potent ABTS radical scavenging activity ($IC_{50}$ = 19.51 ${\mu}g$/mL). Compared with NE#4, relatively lower levels of activity were observed for NE#3 and NE#6, with $IC_{50}$ values of 90.99 and 76.88 ${\mu}g$/mL, respectively. According to results of western blot analysis for determination of SOD expression in $H_2O_2$-treated HepG2 cells, NE#5 and NE#6 induced a dramatic increase in the expression ratio of SOD, compared to the group treated with $H_2O_2$ only. Activity of xanthine oxidase, which converts xanthine into uric acid, generating superoxide ions, was inhibited by NE#4 and NE#6 in a dose-dependent manner. NE#4 induced significant inhibition of mushroom tyrosinase activity. A reduction in cellular melanin contents of 80% was observed in B16F1 melanocytes treated with NE#5 and NE#6; these effects were similar to those of arbutin at 100 ${\mu}M$. In addition, gelatin zymography and reverse transcription-PCR analysis were performed for assessment of anti-photoaging activity of Nuruk. Treatment with NE#6 resulted in dramatically inhibited activities of matrix metalloproteinase (MMP)-2/-9, suppressed expression of MMP-1, and increased expression of type-1 procollagen. Results of gelatin zymography for NE#4 and NE#5 were similar, to a slightly lesser degree. These results suggest the potential of NE#4 and NE#6 as natural ingredients for use in functional foods and cosmetics.

Performance of Crossbred Gilts Fed on Diets with Higher Levels of Fat and Fibre through Addition of Rice Bran

  • Soren, N.M.;Bhar, R.;Chhabra, A.K.;Mandal, A.B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.11
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    • pp.1650-1655
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    • 2003
  • Crossbred gilts (n=54) of about $26.38{\pm}0.85kg$ body weight and 25 weeks of age were randomly divided into 18 groups of 3 animals each. Three dietary treatments viz., $T_1$, $T_2$ and $T_3$ were formulated. The treatment $T_1$ containing maize grain (35%) and wheat bran (47%) along with soybean meal and fish meals served as control diet. Rice bran (RB) was selected as a single source of fat (13.02%) and fibre (17.12%), which was gradually increased in diets $T_2$ (41%) and $T_3$ (82%) replacing maize grain and wheat bran of $T_1$ at 50% and 100% in the diets $T_2$ and $T_3$, respectively. The fat and fibre levels of the diets were thus 3.46 and 5.24, 9.31and 9.69 and 11.61and 13.26% in $T_1$, $T_2$ and $T_3$, respectively. All the diets, however, contained almost similar concentration of CP (18.35${\pm}0.29%$). Each dietary treatment was offered to six replicated groups of 3 piglets in following completely randomized design and feeding was continued for 112 days during the growing phase. Growth, feed utilization, reproductive performance, nutrient utilization and different blood biochemical parameters were studied. Growth rate, feed intake and feed conversion were lower (p<0.01) in $T_3$ in comparison to $T_1$ or $T_2$. Digestibility of all the nutrients except EE was reduced significantly (p<0.01) in $T_3$. Serum glucose level decreased (p<0.01), whereas the urea and cholesterol concentration in the blood increased (p<0.01) in $T_3$. The duration of estrus of pigs fed diet containing 82% RB ($T_3$) was highly variable within the group, but all the groups showed statistically similar duration of estrous. Feed cost per unit gain was found to be comparable between control ($T_1$) and $T_2$ group. The results indicated that RB can be included up to 41% in swine diets replacing 50% of maize and wheat bran. Higher concentrations of ether extract and fibre beyond 9.31 and 9.69% in diet had detrimental effect on growth, nutrient utilization and reproductive performance.

Production of IFN-γ by TNF-α in Macrophages from Tumor Micro Environment; Significance in Angiogenic Switch Control (종양 미세 환경 내 대식세포에서 혈관 신생 조절 인자로서의 TNF-α에 의한 IFN-γ의 분비 조절)

  • Pyo, Suhk-Neung;Baek, Soyoung;Kwak, Jang-Dong;Park, Dae-Sub;Joe, Sung-Jun;Lee, Hyun Ah
    • IMMUNE NETWORK
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    • v.3 no.1
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    • pp.53-60
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    • 2003
  • Background: The role of macrophages in tumor angiogenesis is known to be the production of angiogenic cytokines and growth factors including TNF-${\alpha}$. Recently, macrophage also can produce the INF-${\gamma}$ that is being studied to be involved in angiogenic inhibition. Thus, the importance of macrophages in tumor angiogenesis is might being an angiogenic switch. Thus, the hypothesis tested here is that TNF-${\alpha}$ can modulate the INF-${\gamma}$ production in the macrophages from tumor environment as a part of tumor angiogenic switch. Methods: Macrophages in tumor environment were obtained from the peritoneal cavity of C57BL/6 mice injected with B16F10 melanoma cell line for 6 or 11 days. $Mac1^+$-macrophages were purified using magnetic bead ($MACs^{TM}$; Milteny Biotech, Germany) and cultured with various concentrations of TNF-${\alpha}$ for various time points at $37^{\circ}C$. The supernatants were analyzed for IFN-${\gamma}$ or VEGF by ELISA kit (Endogen, Woburn, MA). Results: Residential macrophages from the peritoneal cavity did not respond to LPS or TNF-${\alpha}$ to produce INF-${\gamma}$. However, the cells from tumor environment produced IFN-${\gamma}$ as well as VEGF and upregulated by the addition of LPS or TNF-${\alpha}$. RT-PCR analysis revealed the external TNF-${\alpha}$-induced IFN-${\gamma}$ gene expression in the macrophages from tumor environment. Conclusion: The overall data suggest that the macrophages in tumor environment might have an important role not only in angiogenic signal but also in anti-angiogenic signal by producing related cytokines. And TNF-${\alpha}$ might be a key cytokine in tumor angiogenic switch.

Study of Skin Depigmenting Mechanism of the Ethanol Extract of Fagopyrum esculentum (교맥 에탄올 추출물의 피부 미백기전 연구)

  • No, Seong-Taek;Kim, Dae-Sung;Lee, Seong-Jin;Park, Dae-Jung;Lee, Jang-Cheon;Lim, Kyu-Sang;Woo, Won-Hong;Mun, Yeun-Ja
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.5
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    • pp.1243-1249
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    • 2007
  • The aim of this study was to investigate the effect of ethanol extract of Fagopyrum esculentum on the melanogenesis. To determine whether ethanol extract of Fagopyrum esculentum suppress melanin synthesis in cellular level, B16F10 melanoma cells were cultured in the presence of different concentrations of Fagopyrum esculentum ethanol extract. In the present study, we examined the effects of Fagopyrum esculentum ethanol extract on cell proliferation, melanin contents, tyrosinase activity, expression of melanogenic enzyme proteins including tyrosinase, tyrosinase-related protein 1 (TRP-1) and tyrosinase-related protein 2 (TRP-2). Cell proliferation was slightly increased by treatment with ethanol extract of Fagopyrum esculentum $(25-200 {\mu}g/m{\ell}).$ The ethanol extract of Fagopyrum esculentum effectively suppressed melanin contents at a dose of $100 {\mu}g/m{\ell}).$ It was observed that the color of cell pellets was totally whitened compared with the control. The ethanol extract of Fagopyrum esculentum inhibited tyrosinase activity, regulate melanin biosynthesis as the key enzyme in melanogenesis. Using western blot analysis, the ethanol extract of Fagopyrum esculentum dose-dependently decreased tyrosinase and TRP-1 protein levels, and tyrosinase and TRP-1 were detected in similar manner. ${\alpha}-MSH$ leads to a stimulation of melanin synthesis through increase of tyrosinase activity, melanin contents and cytoplasmic dendricity. In this study, ethanol extract of Fagopyrum esculentum down-regulated the ${\alpha}-MSH$-induced tyrosinase activity, melanin contents and cytoplasmic dendricity. Regarding protein levels of the melanogenic enzymes, the amounts of tyrosinase and TRP-1 was increased after incubation with a-MSH. The treatment of ethanol extract of Fagopyrum esculentum decreased the ${\alpha}-MSH$-induced expression levels of tyrosinase and TRP-1. These results suggest that the ethanol extract of Fagopyrum esculentum exerts its depigmenting effects through the suppression of tyrosinase, TRP-1 and cytoplasmic dendricity. And it may be a potent depigmetation agent in hyperpigmentation condition.

Analysis of Immunomodulating Gene Expression by cDNA Microarray in $\beta$-Glucan-treated Murine Macrophage

  • Sung, Su-Kyong;Kim, Ha-Won
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2003.11a
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    • pp.98-98
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    • 2003
  • ${\beta}$-(1,3)-D-Glucans have been known to exhibit antitumor and antimicrobial activities. The presence of dectin-1,${\alpha}$, ${\beta}$-glucan receptor of dendritic cell, on macrophage has been controvertial. RT-PCR analysis led to the detection of dectin-1${\alpha}$ and ${\beta}$ in murine macrophage Raw264.7 cell line. Among the various organs of mouse, dectin-1${\alpha}$ and ${\beta}$ were detected in the thymus, lung, spleen, stomach and intestine. To analyze gene expression modulated by ${\beta}$-glucan treated murine Raw264.7 macrophage, total mRNA was applied to cDNA microarray to interrogate the expression of 7,000 known genes. cDNA chip analysis showed that ${\beta}$-glucan of P. osteatus increased gene expressions of immunomodulating genes, membrane antigenic proteins, chemokine ligands, complements, cytokines, various kinases, lectin associated genes and oncogenes in Raw 264.7 cell line. When treated with ${\beta}$-glucan of P. osteatus and LPS, induction of gene expression of TNF-${\alpha}$ and IFN-R1 was confirmed by RT-PCR analysis. Induction of TNF-R type II expression was confirmed by FACS analysis. IL-6 expression was abolished by EDTA in ${\beta}$-glucan and LPS treated Raw264.7 cell line, indicating that ${\beta}$-glucan binds to dectin-l in a Ca$\^$++/ -dependent manner. To increase antitumor efficacy of ${\beta}$-glucan, ginsenoside Rh2 (GRh2) was co-treated with ${\beta}$-glucan in vivo and in vitro tests. IC$\sub$50/ values of GRh2 were 20 and 25 $\mu\textrm{g}$/$m\ell$ in SNU-1 and B16 melanoma F10 cell line, respectively. Co-treatment with ${\beta}$-glucan and GRh2 showed synergistic antitumor activity with cisplatin and mitomycin C both in vitro and in vivo. Single or co-treatment with ${\beta}$-glucan and GRh2 increased tumor bearing mouse life span. Co-treatment with ${\beta}$-glucan and GRh2 showed more increased life span with mitomycin C than that with cisplatin. Antitumor activities were 67% and 72 % by co-injection with ${\beta}$-glucan and GRh2 in the absence or presence of mitomycin C, respectively.

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Effects of Ehwa Makgeolli Containing Oriental Herbs on Skin Whitening and Wrinkles (한방 이화주의 미백 및 피부 주름 개선 효과)

  • Lee, Sang-Jin;Kwon, Yi-Young;Cho, Sung-Won;Kwon, Hee-Suk;Shin, Woo-Chang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.4
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    • pp.550-555
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    • 2013
  • Extrinsic skin aging is characterized by the loss of skin tone and resilience, irregular pigmentation, and deep wrinkles. The aim of this study was to investigate the effects of Ehwa Makgeolli containing oriental herbs (Glycyrrhiza uralensis Fisch., Lycium chinense MILL., Morus alba L., and Saururus chinensis Baill) on skin whitening and wrinkling in human skin cells. We prepared Makgeolli extracts (HEE) with 70% ethanol. HEE significantly inhibited in vitro mushroom tyrosinase activity and reduced the cellular and secreted melanin content of mouse melanoma melanocytes (B16F1 cells). HEE down-regulated the protein expression of tyrosinase related protein (TRP)-1/-2, a key player in melanogenesis. Treatment with HEE in human keratinoctyes (HaCaT cells) inhibited the proteolytic activities of matrix metalloproteinase (MMP)-2/-9 in a dose-dependent manner and dramatically reduced the expression of MMP-2/-9. In addition, HEE attenuated lipopolysaccharide (LPS)-induced nitric oxide production in murine macrophages (RAW264.7 cells). These results indicate that HEE may be a great cosmeceutical ingredient for its whitening, anti-wrinkle, and anti-inflammatory effects.

Effects of Phytase and Carbohydrases Supplementation to Diet with a Partial Replacement of Soybean Meal with Rapeseed Meal and Cottonseed Meal on Growth Performance and Nutrient Digestibility of Growing Pigs

  • Shim, Y.H.;Chae, B.J.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.9
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    • pp.1339-1347
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    • 2003
  • An experiment was conducted to investigate the effects of microbial phytase ($Natuphos^{(R)}$) supplementation in combination with carbohydrases (composed of enzymes targeted to soybean meal (SBM) dietary components such as $\alpha$-galactosides and galactomannans; $Endo-Power^{(R)}$) to corn-soybean meal based diet (CSD) and complex diet (CD) with a partial replacement of SBM with rape seed meal (RSM) and cotton seed meal (CSM) on growth performance and nutrient digestibility of growing pigs. A total of 168 growing pigs averaging $13.18{\pm}1.77kg$ of initial body weight was arranged as a $2{\times}2$ factorial design with main effects of diet types (corn-SBM based diet (CSD) and complex diets (CD; 5% of SBM was replaced with 2.5% of RSM and 2.5% of CSM in diet for phase I (0 to 3 weeks) and 6% of SBM was replaced with 3% of RSM and 3% of CSM in diet for phase II (4 to 7 weeks))) and enzyme supplementation (none and 0.1% of phytase (500 FTU/kg diet) and 0.1% of carbohydrases). The diet with enzyme application were formulated to have a 0.18% unit lower aP than diets without enzyme application. Each treatment had three replicates with 14 pigs per replicate. To determine supplementation effect of phytase and carbohydrases on ileal amino acid digestibility of SBM, RSM and CSM, a total of 18 T-cannulated pigs (initial body weight; $13.52{\pm}1.24kg$) were assigned to six dietary treatments in the present study. Dietary treatments in metabolic trial included 1) SBM diet, 2) SBM diet+with enzymes (phytase (500 FTU/kg) and carbohydrases at 0.1%, respectively), 3) CSM diet, 4) CSM diet+enzymes, 5) RSM diet and 6) RSM diet+enzymes. During whole experimental period (0 to 7 wks), there was no difference in growth performance between diets (CSD and CD). However, dietary phytase and carbohydrases supplementation significantly improved gain/feed ratio (G:F) of growing pigs. During the phase II (4-7 weeks), dietary phytase and carbohydrases supplementation significantly improved all fecal nutrient digestibilities (Dry matter (DM), gross energy (GE), crude protein (CP), crude fat (CF), calcium (Ca) and phosphorus (P)). Dietary phytase and carbohydrases supplementation improved significantly overall ileal amino acid digestibilities of SBM, RSM and CSM based diets (p<0.05). The simultaneous inclusion of phytase and carbohydrases in both of CSD and CD reduced feed cost per kg body weight gain (FCG). Also, results suggest that 2.5 to 3% of RSM and CSM, respectively, might be used as a protein source in growing pig diets without having an adverse effect on the growth performance and nutrient digestibility and simultaneous phytase and carbohydrases addition improves nutritional value of SBM, RSM and CSM by improving ileal amino acid digestibilities.

Effect of 1,2,3,4,6-penta-O-galloyl-$\beta$-D-glucose isolated from Galla Rhois on gap junctional intercellular communication and antiangiogenic activity (오배자 BuOH층에서 분리한 1,2,3,4,6-penta-O-galloyl-$\beta$-D-glucose의 세포간교통 회복 및 혁관형성 억제작용에 대한 효과)

  • Huh Jeong Eun;Lee Hyo Jung;Song Gyu Yong;Cha Bae Cheon;Kim Han Sung;Yoo Dong Youl;Ryu Shi Yong;Kim Sung Hoon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.3
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    • pp.452-457
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    • 2002
  • Galla Rhois is a gallnut of Rhus javanica Linne used for treatment of diarrhea, hemorrhage, cough, leukorrhea and toxic tumor etc in oriental medicine. For the evaluation of antitumor effect of Galla Rhois, activity based fractionation was done. We isolated an effective compound and identified 1,2,3,4,6-penta-O-galloyl-β-D-glucose(PGG) by photometric analysis such as NMR and MASS. Then, we studied the angiogenic activity of PGG. It showed a cytotoxicity against SK-OV-3, SK-OV-3, HT1080 with IC/sub 50/ of 50 ug/ml approximately. It also effectively inhibited proliferation of HUVEC cells treated by bFGF to 30% of control at 20 ug/ml and cell migration to 80% at 10 ug in a dose dependent fashion. Tube formation of HUVEC cells on matrigel was effectively suppressed from 2.5 ug/ml of concentration by PGG. Moreover, it effectively recovered the dysfunction of gap junctional intercellular communication in WB-F344 rat liver epithelial cells caused by hydrogen peroxide at 4 ug/ml suggesting it potently can inhibit tumor promotion. Taken together, it indicates 1,2,3,4,6-penta-O-galloyl- β -D-glucose has antiangiogenic activity.

Effect of Medicinal Herb Prepared through Traditional Antidiabetic Prescription on α-Glucosidase Activity and Evaluation Method for Anti-Melanogenesis Agents Using α-Glucosidase Activity (당뇨 처방에 근거한 생약재의 α-Glucosidase 활성 저해 효과 및 이를 활용한 미백 소재 평가법)

  • Kim, Mi Jin;Im, Kyung Ran;Yoon, Kyung-Sup
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.7
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    • pp.993-999
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    • 2015
  • For the purpose of investigating the in vitro antidiabetic activity of a medicinal herb and herb mixture extracts prepared through traditional antidiabetic prescription, this study examined ${\alpha}$-glucosidase inhibitory activity. Tyrosinase, a type I membrane glycoprotein, is synthesized and glycosylated in the endoplasmic reticulum (ER) and Golgi. The enzyme is subsequently transported to melanosomes, where it participates in melanogenesis. Previous studies showed that disruption of early ER N-glycan processing by an ${\alpha}$-glucosidase inhibitor suppresses tyrosinase enzymatic activity and melanogenesis. According to the results, most oriental medicinal herbal extracts were stronger than acarbose and N-butyldeoxynojirimycin, known as an ${\alpha}$-glucosidase inhibitor. Interestingly, ethyl acetate layer of enzyme hydrolyzed Cheongsimyeonjaeum had an inhibitory effect on melanin synthesis in B16F1 cells, although it did not inhibit tyrosinase activity directly. Together, ${\alpha}$-glucosidase inhibition activity could be used to evaluate anti-melanogenesis, although cross-checking with melanin inhibitory assay is recommended.

Studies on Direct Sowing-Dry Paddy Rice Culture in the Middle Part of Korea (중부지방에 있어서의 수도건답직파재배 기술체계확립에 관한 시험연구)

  • Jai-Hyoun Lee
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.7 no.1
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    • pp.1-29
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    • 1969
  • Experiments on rice concerning it s varieties, fertilization, seedling dates and herbicides have been conducted to determine the most desirable method of direct sowing cultivation on dry paddy field land in the middle part of Korea. The results obtained at the Office of Rural Development of Choongnam Province are as follows:. 1. Sixteen different derivatives from the main varieties of low land rice were cultivated on a dry paddy field by the direct sowing method; at the same time, a few varieties were tried adopting the common transplanting cultivation method. The yield and yield factors from these two groups were examined to give the following results: a) Compared to the common transplanting cultivation, the direct sowing method showed remarkably increased number of panicles while the number of flowers per panicle was shown to be significantly decreased. The maturing ratio was detected to be lowered. The yield horn them differed according to the different varieties : good yield was obtained from Hokwang while Norin #25 proved poor when compared with the common transplanting cultivation method. b) Among sixteen varieties tested, Sunsou, Norin #25, Jaigou, Hokwang, Palkueng and Gosi showed comparatively high yields, their yield being more than 325 kilograms per 10 Are, but Nampoong, Paldal, Nongkwang, Norin #29, Eunbangju #101 and Shiro gane showed less yield, their yield being less than 271 kilograms per 10 Are, the relations between the yield and the yield factors can be summarized as follows; Number of varieties and their rice yield. 1) The varieties which were great in the, number of panicles and high in yield=Jaigoun, Hokwang Palkueng and Gosi. 2) The varieties which were low in the number of panicles and high in yield=Sounsou and Norin #25. 3) The varieties which were great in the number of panicles and poor in yield=Eunbangju #101 and Sirogane. 4) The varieties which were poor in the number of panicles and poor in yield: Nampung, Paldal and Norin #29. Number of flowers per panicle and yield. 1) The varieties which were great in the number of flowers per panicle and high in yield: Sounsou, Norin #25 and Gosi. 2) The varieties which were poor in the number of flowers per panicle and high in yield ; Jaigoun, Hokwang and Palkueng. 3) The varieties which were great in the number of flowers per panicle and poor in yield: Paldal and Nampung. 4) The varieties which were poor in the number of flowers per panicle and poor in yield: Norin #29. Eunbangju #101 and Sirogane. Maturing ratio and yield. 1) The varieties which were high in the maturing ratio and high in yield: Jaigoun, Sounsou, Norin #25 and Palkueng. 2) The varieties which were low in the maturing ratio and high in yield: Hokwang and Gosi. 3) The varieties which were early maturing rat io and low in yield: Hokwang and Gosi. 4) The varieties which were late maturing ratio and poor in yield: Eunbangju #101, Nampungand Sirogane 1, 000 grain weight and yield. 1) The varieties which were heavy in 1, 000 grains weight and high in yield=Norin #25 and Hokwang. 2) The varieties which were light in 1, 000 grains weight and high in yield=Sounsou and Jaigoun. 3) The varieties which were heavy in 1, 000 grains weight and poor in yield=Nongkwang and Eunbanju. 4) The varieties which were light in 1, 000 grains weight and poor in yield=Norin #29 and Sirogane. 2. The experiment on fertilization showed that the most desirable amount to be given per 10 Are was 10 kilograms of Nitrogen, 5 kilograms of phosphate and 6 kilograms of potassium; and when the Nitrogen given exceeded 8 kilograms, its effect was better when given in amsll consecutive (split) amounts, while the maturing ratio and the number of the flowers per panicle increased when Nitrogen was given in large amount during the later stage of growth of rice. 3. The experiment on the date and amount of seedling showed that the tested variety, Sunsou gave the best results when planted on the days between 25 April and 10 May. Eight liters per 10 Are were preferable if planted early and 12 liters per 10 Are if planted late. The reason why the later planting gave a lower yield was that the number of flowers per panicle was fewer. 4. The experiment on the irrigation for rice with direct sowing cultivation immersed in water showed that it was the most satisfactory when irrigated on 25th June, 55 days after its seedling, its plot giving the best yield. The plots 10th June and 15th July showed just as good results. However, irrigated later, than 15th July it showed lower yields. 5. Compared to the yield of the plot controlled by the common method, the yield from the plots treated with chemical herbicide such as LOROX, TOK, PCP, SWEP, Mo-338 on dry condition soil seemed poorer, but significant difference was not found statistically. On the other hand in the case where chemical herbicides such as TOK, Mo-338, Stam F-34 or ORDRAM were used after irrigation, the yield from the ORDRAM and TOK treated plots did not show significant differences compared to the common hand weed controling method, but those treated with chemicals other than the above showed a lower yield.

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