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Development and Validation of the Analytical Method for Oxytetracycline in Agricultural Products using QuEChERS and LC-MS/MS (QuEChERS법 및 LC-MS/MS를 이용한 농산물 중 Oxytetracycline의 잔류시험법 개발 및 검증)

  • Cho, Sung Min;Do, Jung-Ah;Lee, Han Sol;Park, Ji-Su;Shin, Hye-Sun;Jang, Dong Eun;Cho, Myong-Shik;Jung, ong-hyun;Lee, Kangbong
    • Journal of Food Hygiene and Safety
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    • v.34 no.3
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    • pp.227-234
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    • 2019
  • An analytical method was developed for the determination of oxytetracycline in agricultural products using the QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method by liquid chromatography-tandem mass spectrometry (LC-MS/MS). After the samples were extracted with methanol, the extracts were adjusted to pH 4 by formic acid and sodium chloride was added to remove water. Dispersive solid phase extraction (d-SPE) cleanup was carried out using $MgSO_4$ (anhydrous magnesium sulfate), PSA (primary secondary amine), $C_{18}$ (octadecyl) and GCB (graphitized carbon black). The analytes were quantified and confirmed with LC-MS/MS using ESI (electrospray ionization) in positive ion MRM (multiple reaction monitoring) mode. The matrix-matched calibration curves were constructed using six levels ($0.001{\sim}0.25{\mu}g/mL$) and coefficient of determination ($r^2$) was above 0.99. Recovery results at three concentrations (LOQ, $10{\times}LOQ$, and $50{\times}LOQ$, n=5) were from 80.0 to 108.2% with relative standard deviations (RSDs) less than of 11.4%. For inter-laboratory validation, the average recovery was in the range of 83.5~103.2% and the coefficient of variation (CV) was below 14.1%. All results satisfied the criteria ranges requested in the Codex guidelines (CAC/GL 40-1993, 2003) and the Food Safety Evaluation Department guidelines (2016). The proposed analytical method was accurate, effective and sensitive for oxytetracycline determination in agricultural commodities. This study could be useful for safety management of oxytetracycline residues in agricultural products.

Study on In Vitro Aggregation and Culture of Mouse Embryos by Phytohemagglutinin-P (Phytohemagglutinin-P 첨가(添加)에 따른 생쥐배(胚)의 시험관내(試驗管內) 응집(凝集)과 배양(培養)에 관하여)

  • Park, Hang Kyun;Ryou, Zae Yoong
    • Current Research on Agriculture and Life Sciences
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    • v.7
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    • pp.83-97
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    • 1989
  • This study was carried out to obtain basic information necessary for aggregation and in-vitro culture of mouse embryos by treating phytohemagglutinin-p (PHA-P). The 4-, 8-cell and morula embryos were obtained from female mice of albino BALE/C, CBA and C57BL strains, those were injected 5 i.u pregenant mare serum gonadotrophin and 5 i.u human chorionic gonadotrophin to superovulation. The zona pellucidia was removed by placing the embryos in Acidic Tyrode solution containing 1.0% protease or/and 5 ug/ml PHA-P. The pairs of zona free embryos were subjected to aggregation by glassneedle in BMOC-3 containing 5 ug/ml PHA-P. The aggregation embryos were cultured in Brinster's mouse ova culture-3(BMOC-3) medium under the gas phase of 5% $CO_2$ in air $37^{\circ}C$ for 13 to 50 hours. The results obtained in this study are summarised as follows : 1. When 4-, 8-cell and morula embryos were zona-freed in acidic Tyrode solution containing 1.0% protease or/and 5 ug/ml PHA-P, and cultured in vitro to blastocysts, the 4- and 8-cell embryos showed slightly less development rates than the morula one did, and solution of 5 ug/ml PHA-P brought some higher development rate than negative control. 2. As 2, 5 or 10 ug/ml PHA-P was added to the solution to aggregate 4-, 8-cell or morula embryos, 2 ug/ml solution represented slightly lower aggregation rate than the higher levels solutions, and 4- and 8-cell embryos showed higher rates than morula one did (P<.05). 3. In respect to the development rates of aggregated embryos to morula no significant difference was found among PHA-P levels and between 4-and 8-cell embryos. With respect to those of aggregated embryos to blastocysts the different levels of PHA-P showed similar results, however, the 4- and 8-cell embryos represented higher rates than the morula one did (P<.05). 4. The mean time necessary for development of aggregated 4-, 8-cell and morula embryos to blastocysts were 38.5-40, 26-27 and 19-20hrs. Respectively in solution for aggregation. 5. The aggregation rates of embryos were 34-94%, when treated protease or/and PHA-P. Supplementation of 5 ug/ml PHA-P to the solution for aggregation showed a trend demonstrating higher aggregation rate compared to negative control, although no significance was found. However, 4- and 8-cell embryos represented significantly higher aggregation rates than the morula one did (P<.05). 6. The development rates of 4- and 8-cell embryos to morula were 52.7-84.7 and 73.8-87.2%, respectively, showing no significant difference between two cell stages. However, the aggregation rates of embryos treated with solution containing PHA-P were higher than negative control (P<.05). 7. The development rates of 4- and 8-cell and morula embryos to blastocysts were 41.7-77.7 78.7-83.0 and 0-19.2%, respectively. The rates of 4-cell embryos treated with PHA-P were significant higher than the negative control (P<.05). The 8-cell and morula embryos also showed more rates when treated PHA-P.

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Effects of NG-monomethyl-L-arginine and L-arginine on cerebral hemodynamics and energy metabolism during reoxygenation-reperfusion after cerebral hypoxia-ischemia in newborn piglets (급성 저산소성 허혈성 뇌손상이 유발된 신생자돈에서 재산소-재관류기 동안 NG-monomethyl-L-arginine과 L-arginine이 뇌의 혈역학 및 에너지 대사에 미치는 영향)

  • Ko, Sun Young;Kang, Saem;Chang, Yun Sil;Park, Eun Ae;Park, Won Soon
    • Clinical and Experimental Pediatrics
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    • v.49 no.3
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    • pp.317-325
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    • 2006
  • Purpose : This study was carried out to elucidate the effects of nitric oxide synthase(NOS) inhibitor, NG-monomethyl-L-arginine(L-NMMA) and nitric oxide precursor, L-arginine(L-Arg) on cerebral hemodynamics and energy metabolism during reoxygenation-reperfusion(RR) after hypoxia-ischemia(HI) in newborn piglets. Methods : Twenty-eight newborn piglets were divided into 4 groups; Sham normal control(NC), experimental control(EC), L-NMMA(HI & RR with L-NMMA), and L-Arg(HI & RR with L-Arg) groups. HI was induced by occlusion of bilateral common carotid arteries and simultaneously breathing with 8 percent oxygen for 30 mins, and followed RR by release of carotid occlusion and normoxic ventilation for one hour. All groups were monitored with cerebral hemodynamics and cytochrome $aa_3$ (Cyt $aa_3$) using near infrared spectroscopy(NIRS). $Na^+$, $K^+$-ATPase activity, lipid peroxidation products, and tissue high energy phosphate levels were determined biochemically in the cerebral cortex. Results : In experimental groups, mean arterial blood pressure, $PaO_2$, and pH decreased, and base excess and blood lactate level increased after HI compared to NC group(P<0.05). These variables subsequently returned to baseline after RR except pH. There were no differences among the experimental groups. In NIRS, oxidized hemoglobin($HbO_2$) decreased and hemoglobin(Hb) increased during HI(P<0.05) but returned to base line immediately after RR; 40 min after RR, the $HbO_2$ had decreased significantly compared to NC group(P<0.05). Changes of Cyt $aa_3$ decreased significantly compared to NC after HI and recovered at the end of the experiment. Significantly reduced cerebral cortical cell membrane $Na^+$, $K^+$-ATPase activity and increased lipid peroxidation products(P<0.05) were not improved with L-NMMA or L-Arg. Conclusion : These findings suggest that NO is not involved in the mechanism of HI and RR brain damage during the early acute phase of RR.

A Study on the Cultivation Processes and Settlement Developments on the Mangyoung River Valley (만경강유역의 개간과정과 취락형성발달에 관한 연구)

  • NamGoong, Bong
    • Journal of the Korean association of regional geographers
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    • v.3 no.2
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    • pp.37-87
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    • 1997
  • As a results of researches on the cultivation processes and settlement developments on the Mangyoung river valley as a whole could be have four 'Space-Time Continuity' through a [Origin-Destination] theory model. On a initial phases of cultivation, the cultivation process has been begun at mountain slopes and tributory plains in upper part of river-basin from Koryo Dynasty to early Chosun Dynasty. At first, indigenous peasants burned forests on the mountain slopes for making 'dryfield' for a cereal crops. Following population increase more stable food supply is necessary facets of life inducing a change production method into a 'wetfield' in tributory plains matching the population increase. First sedentary agriculture maybe initiated at this mountain slopes and tributory plains on upper part of river basin through a burning cultivation methods. Mountain slopes and tributory plains are become a Origin area in cultivation processes. It expanded from up to down through the valleys with 'a bits of land' fashion in a steady pace like a terraced fields expanded with bit by bit of land to downward. They expanded their land to the middle part of river basin in mid period of Chosun Dynasty with dike construction techniques on the river bank. Lower part of river cultivated with embankment building techniques in 1920s and then naturally expanded to the tidal marshes on the estuaries and river inlets of coastal areas. 'Pioneer fringes' are consolidated at there in modern times. Changes in landscapes are appeared it's own characters with each periods of time. Followings are results of study through the Mangyoung river valley as a whole. (1) Mountain slopes and tributory plains on the upper part of river are cultivated 'dryfields' by indigenous peasants with Burning cultivation methods at first and developed sedentary settlements at the edges of mountain slopes and on the river terrace near the fields. They formed a kind of 'periphery-located cluster type' of settlement. This type of settlement are become a prominant type in upper part of river basin. 'Dryfields' has been changed into a 'wetfields' at the narrow tributory plains by increasing population pressure in later time. These wetfields are supplied water by Weir and Ponds Irrigation System(제언수리방법). Streams on the tributory plains has been attracted wetfields besides of it and formed a [water+land] complex on it. 'Wetfields' are expanded from up to downward with a terraced land pattern(adder like pattern, 붕전) according to the gradient of valley. These periphery located settlements are formed a intimate ecological linkage with several sets of surroundings. Inner villages are expanded to Outer villages according to the expansion of arable lands into downward. (2) Mountain slopes and tributory plains expanded its territory to the alluvial deposited plains on the middle part of river valley with a urgent need of new land by population increase. This part of alluvial plains are cultivated mainly in mid period of Chosun Dynasty. Irrigation methods are changed into a Dike Construction Irrigation method(천방수리방법) for the control of floods. It has a trend to change the subjectives of cultivation from community-oriented one who constructed Bochang along tributories making rice paddies to local government authorities who could be gather large sums of capitals, techniques and labours for the big dike construction affairs. Settlements are advanced in the midst of plains avoiding friction of distances and formed a 'Centrallocated cluster type' of settlements. There occured a hierarchical structures of settlements in ranks and sizes according merits of water supply and transportation convenience at the broad plains. Big towns are developed at there. It strengthened a more prominant [water+land] complex along the canals. Ecological linkages between settlements and surroundings are shaded out into a tiny one in this area. (3) It is very necessary to get a modern technology of flood control at the rivers that have a large volume of water and broad width. The alluvial plains are remained in a wilderness phase until a technical level reached a large artificial levee construction ability that could protect the arable land from flood. Until that time on most of alluvial land at the lower part of river are remained a wilderness of overgrown with reeds in lacks of techniques to build a large-scale artificial levee along the riverbank. Cultivation processes are progressed in a large scale one by Japanese agricultural companies with [River Rennovation Project] of central government in 1920s. Large scale artificial levees are constructed along the riverbank. Subjectives of cultivation are changed from Korean peasants to Japanese agricultural companies and Korean peasants fell down as a tenant in a colonial situation of that time in Korea. They could not have any voices in planning of spatial structure and decreased their role in planning. Newly cultivated lands are reflected company's intensions, objectives and perspectives for achieving their goals for the sake of colonial power. Newly cultivated lands are planned into a regular Rectangular Block settings of rice paddies and implanted a large scale Bureaucratic-oriented Irrigation System on the cultivated plains. Every settlements are located in the midst of rice paddies with a Central located Cluster type of settlements. [water+land] complex along the canal system are more strengthened. Cultivated space has a characters of [I-IT] landscapes. (4) Artificial levees are connected into a coastal emnankment for a reclamation of broad tidal marshes on the estuaries and inlets of rivers in the colonial times. Subjectives of reclamation are enlarged into a big agricultural companies that could be acted a role as a big cultivator. After that time on most of reclamation project of tidal marshes are controlled by these agricultural companies formed by mostly Japanese capitalists. Reclaimed lands on the estuaries and river inlets are under hands of agricultural companies and all the spatial structures are formed by their intensions, objectives and perspectives. They constructed a Unit Farming Area for the sake of companies. Spatial structures are planned in a regular one with broad arable land for the rice production of rectangular blocks, regular canal systems and tank reservoir for the irrigation water supply into reclaimed lands. There developed a 'Central-located linear type' of settlements in midst of reclaimed land. These settlements are settled in a detail program upon this newly reclaimed land at once with a master plan and they have planned patterns in their distribution, building materials, location, and form. Ecological linkage between Newly settled settlemrnts and its surroundings are lost its colours and became a more artificial one by human-centred environment. [I-IT] landscapes are become more prominant. This region is a destination area of [Origin-Destination] theory model and formed a 'Pioneer Fringe'. It is a kind of pioneer front that could advance or retreat discontinously by physical conditions and socio-cultural conditions of that region.

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A Randomized, Double-Blind Study to Assess the Efficacy and Safety of Oral LB20304 (Gemifloxacin) at Doses of 160mg or 320mg (Equivalent to 200mg or 400mg of the Mesylate Salt) Once Daily for 7 Days for the Treatment of Acute Exacerbations of Chronic Bronchitis (AECB) in Korean Adult Population (한국인에서 만성기관지염의 급성악화를 치료하기 위한 LB20304(Gemifloxacin) 160mg 또는 320mg 1일 1회 7일간 투여의 유효성과 안전성에 대한 연구)

  • Kim, Young Whan;Shim, Young-Soo;Kim, Won Dong;Shim, Tae Sun;Kang, Hong Mo;Choi, ByoungWhui;Kim, Jae-Yeol;Kwon, O Jung;Kim, Hojoong;Kim, Ju Ock;Jung, Ki-Suck;Hyeon, In Gyou;Mo, Eun Kyung;Lee, Seung-Joon;Nam, Gui Hyun;Lee, Kye Young;Park, Jae Seuk
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.1
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    • pp.69-87
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    • 2003
  • Background : LB20304(gemifloxacin) is a new fluoroquinolone antibacterial agent with excellent activity against both Gram-negative and Gram-positive organisms. In vitro studies using clinical isolates have shown gemifloxacin to be highly active against penicillin-resistant strains of S. pneumoniae and in contrast to other reference quinolones, gemifloxacin retained good activity against clinical isolates of S. pneumoniae that were resistant to other members of the quinolone class. Therefore, gemifloxacin is thought to be effective in treating acute bacterial exacerbation of chronic bronchitis(AECB). The objective of this study was to evaluate the efficacy and safety of oral gemifloxacin at doses of 160mg or 320mg once daily for 7 days for the treatment of AECB in Korean adult population. Methods : This was a randomized, multicenter, double-blind, parallel group Phase II study to assess the clinical and antibacterial efficacy and safety of oral gemifloxacin for the treatment of AECB. Treatment Group A (67 patients) took oral gemifloxacin 160mg once daily for seven days and treatment Group B (70 patients) took oral gemifloxacin 320mg once daily for seven days. Results : The demographic profiles of the two treatment groups were similar. The clinical response at follow-up was 84.2% in the gemifloxacin 160-mg group, and 88.7% in the gemifloxacin-320 mg group, showing no statistically significant difference between two treatment groups(p=0.49). The clinical response at the end of therapy was 96.5% in the 160-mg group, and 96.4% in the 320-mg group. The bacteriological response at the end of therapy and follow-up were 81.8% and 78.9%, respectively, in the 160-mg group, and 86.4% and 84.2%, respectively, in the 320-mg group, showing no statistically significant difference between two treatment groups(p=0.68 and 0.68, respectively). S. pneumoniae(12 isolates) and H. influenzae(10 isolates) were the most prevalent pathogens. The MICs were lower for gemifloxacin than other quinolones against these key pathogens, and for S. pneumoniae, the MICs for gemifloxacin were considerably lower(${\leq}0.03$ ug/mL) than those for other quinolones, beta-lactams and macrolides. In the period on-therapy plus 30 days post-therapy, a total of 18 patients(26.9%) in the gemifloxacin 160mg group and 22 patients(31.4%) in the 320mg group reported at least one adverse event(AE). The most frequently reported AE was abdominal pain(3/67 patients, 4.5%) in the gemifloxacin 160mg group and increased level of hepatic enzyme(5/70 patients, 7.1%) in the 320mg group. The overall AE profiles for the two treatment groups were similar. Two out of 67 patients(3.0%) in the gemifloxacin 160mg group and 1/70 patients(1.4%) in the 320mg group reported at least one serious AE, however, none of which was considered by the investigator to be of suspected or probable relationship to study medication. Conclusion : The results of this study showed that gemifloxacin at doses of 160mg or 320mg once daily for 7 days in the treatment of acute exacerbations of chronic bronchitis(AECB) in adult Koreans was a very effective and safe treatment both clinically and bacteriologically.

Studies on the Varietal Difference in the Physiology of Ripening in Rice with Special Reference to Raising the Percentage of Ripened Grains (수도 등숙의 품종간차이와 그 향상에 관한 연구)

  • Su-Bong Ahn
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.14
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    • pp.1-40
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    • 1973
  • There is a general tendency to increase nitrogen level in rice production to insure an increased yield. On the other hand, percentage of ripened grains is getting decreased with such an increased fertilizer level. Decreasing of the percentage is one of the important yield limiting factors. Especially the newly developed rice variety, 'Tongil' is characterized by a relatively low percentage of ripened grains as compared with the other leading varieties. Therefore, these studies were aimed to finding out of some measures for the improvement of ripening in rice. The studies had been carried out in the field and in the phytotron during the period of three years from 1970 to 1972 at the Crop Experiment Station in Suwon. The results obtained from the experiments could be summarized as follows: 1. The spikelet of Tongil was longer in length, more narrow in width, thinner in thickness, smaller in the volume of grains and lighter in grain weight than those of Jinheung. The specific gravity of grain was closely correlated with grain weight and the relationship with thickness, width and length was getting smaller in Jinheung. On the other hand, Tongil showed a different pattern from Jinheung. The relationship of the specific gravity with grain weight was the greatest and followed by that with the width, thickness and length, in order. 2. The distribution of grain weight selected by specific gravity was different from one variety to another. Most of grains of Jinheung were distributed over the specific gravity of 1.12 with its peak at 1.18, but many of grains of Tongil were distributed below 1.12 with its peak at 1.16. The brown/rough rice ratio was sharply declined below the specific gravity of 1.06 in Jinheung, but that of Tongil was not declined from the 1.20 to the 0.96. Accordingly, it seemed to be unfair to make the specific gravity criterion for ripened grains at 1.06 in the Tongil variety. 3. The increasing tendency of grain weight after flowering was different depending on varieties. Generally speaking, rice varieties originated from cold area showed a slow grain weight increase while Tongil was rapid except at lower temperature in late ripening stage. 4. In the late-tillered culms or weak culms, the number of spikelets was small and the percentage of ripened grains was low. Tongil produced more late-tillered culms and had a longer flowering duration especially at lower temperature, resulting in a lower percentage of ripened grains. 5. The leaf blade of Tongil was short, broad and errect, having light receiving status for photosynthesis was better. The photosynthetic activity of Tongil per unit leaf area was higher than that of Jinheung at higher temperature, but lower at lower temperature. 6. Tongil was highly resistant to lodging because of short culm length, and thick lower-internodes. Before flowering, Tongil had a relatively higher amount of sugars, phosphate, silicate, calcium, manganese and magnesium. 7. The number of spikelets of Tongil was much more than that of Jinheung. The negative correlation was observed between the number of spikelets and percentage of ripened grains in Jinheung, but no correlation was found in Tongil grown at higher temperature. Therefore, grain yield was increased with increased number of spikelets in Tongil. Anthesis was not occurred below 21$^{\circ}C$ in Tongil, so sterile spikelets were increased at lower temperature during flowering stage. 8. The root distribution of Jinheung was deeper than that of Tongil. The root activity of Tongil evaluated by $\alpha$-naphthylamine oxidation method, was higher than that of Jinheung at higher temperature, but lower at lower temperature. It is seemed to be related with discoloration of leaf blades. 9. Tongil had a better light receiving status for photosynthesis and a better productive structure with balance between photosynthesis and respiration, so it is seemed that tongil has more ideal plant type for getting of a higher grain yield as compared with Jinheung. 10. Solar radiation during the 10 days before to 30 days after flowering seemed enough for ripening in suwon, but the air temperature dropped down below 22$^{\circ}C$ beyond August 25. Therefore, it was believed that air temperature is one of ripening limiting factors in this case. 11. The optimum temperature for ripening in Jinheung was relatively lower than that of Tongil requriing more than $25^{\circ}C$. Air temperature below 21$^{\circ}C$ was one of limiting factors for ripening in Tongil. 12. It seemed that Jinheung has relatively high photosensitivity and moderate thermosensitivity, while Tongil has a low photosensitivity, high thermosensitivity and longer basic vegetative phase. 13. Under a condition of higher nitrogen application at late growing stage, the grain yield of Jinheung was increased with improvement of percentage of ripened grains, while grain yield of Tongil decreased due to decreasing the number of spikelets although photosynthetic activity after flowering was. increased. 14. The grain yield of Jinheung was decreased slightly in the late transplanting culture since its photosynthetic activity was relatively high at lower temperature, but that of Tonil was decreased due to its inactive photosynthetic activity at lower temperature. The highest yield of Tongil was obtained in the early transplanting culture. 15. Tongil was adapted to a higher fertilizer and dense transplanting, and the percentage of ripened grains was improved by shortening of the flowering duration with increased number of seedlings per hill. 16. The percentage of vigorous tillers was increased with a denser transplanting and increasing in number of seedlings per hill. 17. The possibility to improve percentage of ripened grains was shown with phosphate application at lower temperature. The above mentioned results are again summarized below. The Japonica type leading varieties should be flowered before August 20 to insure a satisfactory ripening of grains. Nitrogen applied should not be more than 7.5kg/10a as the basal-dressing and the remained nitrogen should be applied at the later growing stage to increase their photosynthetic activity. The morphological and physiological characteristics of Tongil, a semi-dwarf, Indica $\times$ Japonica hybrid variety, are very different from those of other leading rice varieties, requring changes in seed selection by specific gravity method, in milling and in the cultural practices. Considering the peculiar distribution of grains selected by the method and the brown/rough rice ratio, the specific gravity criterion for seed selection should be changed from the currently employed 1.06 to about 0.96 for Tongil. In milling process, it would be advisable to bear in mind the specific traits of Tongil grain appearance. Tongil is a variety with many weak tillers and under lower temperature condition flowering is delayed. Such characteristics result in inactivation of roots and leaf blades which affects substantially lowering of the percentage of ripened grains due to increased unfertilized spikelets. In addition, Tongil is adapted well to higher nitrogen application. Therefore, it would be recommended to transplant Tongil variety earlier in season under the condition of higer nitrogen, phosphate and silicate. A dense planting-space with three vigorous seedlings per hill should be practiced in this case. In order to manifest fully the capability of Tongil, several aspects such as the varietal improvement, culural practices and milling process should be more intensively considered in the future.he future.

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Time Course Variation of Vitamin $C_3$ Content in Leg Skin of Broiler Chicks Exposed to Different Dose of UVB Light (자외선의 상이한 선양을 조사한 브로일러 병아리의 다리 피부중 비타민 $C_3$ 함양의 경시적 변화)

  • 장윤환;김강수;여영수;강훈석;조인호;배은경
    • Korean Journal of Poultry Science
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    • v.20 no.2
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    • pp.93-105
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    • 1993
  • This study was carried out to determine the concentrations of previtamin D$_3$(PreD$_3$), lumisterol$_3$(L3), tachystero1$_3$(73), vitamin D$_3$(VD$_3$) and provitamin D$_3$(ProD$_3$) in leg skins of broiler chicks exposed to UVB lights (maximum intensity at 297 nm) with dose of 0.204 or 0.409 mJ/$\textrm{cm}^2$(30 or 60 min irradiation) . The broiler Hubbard line day old chicks(2 dose $\times$9 elapsed time $\times$4 replica+10 control=82) were fed VD-deficient diet for 31 days in a windowless subdued light room. The skin was collected at 0, 6, 12, 18, 30, 42, 66, 90 or 138 hr after UVB irradiation. The skin lipid was extracted by 9% ethyl acetate/n-hexane, and the fraction of VD$_3$ and its analogues was purified by Sep-Pak silica cartridge. The straight phase HPLC was utilized to analyze ProD$_3$ and its products. The mole %(absolute level expressed in ng/$\textrm{cm}^2$) of PreD$_3$ in leg skin (epidermis+dermis) was 4.67%(44 ng/$\textrm{cm}^2$) or 3.97%(37 ng/$\textrm{cm}^2$) right after UVB irradiation by 0.204 or 0.409 mJ/$\textrm{cm}^2$(30 or 60 min) at 15 cm distance, respectively. It content in leg skin at 0 hr after exposure was 7.24%(12 ng/$\textrm{cm}^2$) or 0.92%(9 ng/$\textrm{cm}^2$), respectively. The increase in irradiation dose did not affect proportionally the If synthesis.73 concentration in leg skin was 0.58%(S ng/$\textrm{cm}^2$) or 0.57%(6 ng/$\textrm{cm}^2$), respectively 0 hr after irradiation. The VD$_3$ in leg skin of birds exposed to UVB light with dose of 0.204 or 0.409 mJ/$\textrm{cm}^2$ was 2.13% (21 ng/$\textrm{cm}^2$) or 0.97% (16ng/$\textrm{cm}^2$), respectively at 0 hr after exposure, 2.72%(26ng/$\textrm{cm}^2$) or 3.84%(37ng/$\textrm{cm}^2$), respectively at 6 hr, and 4.30% ((33ng/$\textrm{cm}^2$) or 6.40%(76ng/$\textrm{cm}^2$), respectively at 12 hr. The peak concentration of VD$_3$ was presented at 18 or 30 hr when 0.204 or 0.409 mJ/$\textrm{cm}^2$) was treated, respectively. It was shown that 18~30 hr were necessary for the thermal conversion of PreD$_3$ into VD$_3$ in the leg skin of broiler chicks. The ProD$_3$ contents in leg skins of negative control, 0.204 mJ/$\textrm{cm}^2$ and 0.409 mJ/$\textrm{cm}^2$ treated birds were 966, 948 and 815 ng/$\textrm{cm}^2$, respectively at right before and after UVB exposure. It was estimated that 18 or 151 ng/$\textrm{cm}^2$ of ProD$_3$ was isomerized to PreD$_3$, L$_3$, T$_3$ and VD$_3$ when exposed to 0.204 or 0.409 mJ/$\textrm{cm}^2$, respective)y. Consequently it was shown that when double dose of UVB light was applied to irradiate the chick body, more but not double synthesis of VD$_3$ and its analogues was occured in leg skin of brolier chicks.

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