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Field Studios of In-situ Aerobic Cometabolism of Chlorinated Aliphatic Hydrocarbons

  • Semprini, Lewts
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.04a
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    • pp.3-4
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    • 2004
  • Results will be presented from two field studies that evaluated the in-situ treatment of chlorinated aliphatic hydrocarbons (CAHs) using aerobic cometabolism. In the first study, a cometabolic air sparging (CAS) demonstration was conducted at McClellan Air Force Base (AFB), California, to treat chlorinated aliphatic hydrocarbons (CAHs) in groundwater using propane as the cometabolic substrate. A propane-biostimulated zone was sparged with a propane/air mixture and a control zone was sparged with air alone. Propane-utilizers were effectively stimulated in the saturated zone with repeated intermediate sparging of propane and air. Propane delivery, however, was not uniform, with propane mainly observed in down-gradient observation wells. Trichloroethene (TCE), cis-1, 2-dichloroethene (c-DCE), and dissolved oxygen (DO) concentration levels decreased in proportion with propane usage, with c-DCE decreasing more rapidly than TCE. The more rapid removal of c-DCE indicated biotransformation and not just physical removal by stripping. Propane utilization rates and rates of CAH removal slowed after three to four months of repeated propane additions, which coincided with tile depletion of nitrogen (as nitrate). Ammonia was then added to the propane/air mixture as a nitrogen source. After a six-month period between propane additions, rapid propane-utilization was observed. Nitrate was present due to groundwater flow into the treatment zone and/or by the oxidation of tile previously injected ammonia. In the propane-stimulated zone, c-DCE concentrations decreased below tile detection limit (1 $\mu$g/L), and TCE concentrations ranged from less than 5 $\mu$g/L to 30 $\mu$g/L, representing removals of 90 to 97%. In the air sparged control zone, TCE was removed at only two monitoring locations nearest the sparge-well, to concentrations of 15 $\mu$g/L and 60 $\mu$g/L. The responses indicate that stripping as well as biological treatment were responsible for the removal of contaminants in the biostimulated zone, with biostimulation enhancing removals to lower contaminant levels. As part of that study bacterial population shifts that occurred in the groundwater during CAS and air sparging control were evaluated by length heterogeneity polymerase chain reaction (LH-PCR) fragment analysis. The results showed that an organism(5) that had a fragment size of 385 base pairs (385 bp) was positively correlated with propane removal rates. The 385 bp fragment consisted of up to 83% of the total fragments in the analysis when propane removal rates peaked. A 16S rRNA clone library made from the bacteria sampled in propane sparged groundwater included clones of a TM7 division bacterium that had a 385bp LH-PCR fragment; no other bacterial species with this fragment size were detected. Both propane removal rates and the 385bp LH-PCR fragment decreased as nitrate levels in the groundwater decreased. In the second study the potential for bioaugmentation of a butane culture was evaluated in a series of field tests conducted at the Moffett Field Air Station in California. A butane-utilizing mixed culture that was effective in transforming 1, 1-dichloroethene (1, 1-DCE), 1, 1, 1-trichloroethane (1, 1, 1-TCA), and 1, 1-dichloroethane (1, 1-DCA) was added to the saturated zone at the test site. This mixture of contaminants was evaluated since they are often present as together as the result of 1, 1, 1-TCA contamination and the abiotic and biotic transformation of 1, 1, 1-TCA to 1, 1-DCE and 1, 1-DCA. Model simulations were performed prior to the initiation of the field study. The simulations were performed with a transport code that included processes for in-situ cometabolism, including microbial growth and decay, substrate and oxygen utilization, and the cometabolism of dual contaminants (1, 1-DCE and 1, 1, 1-TCA). Based on the results of detailed kinetic studies with the culture, cometabolic transformation kinetics were incorporated that butane mixed-inhibition on 1, 1-DCE and 1, 1, 1-TCA transformation, and competitive inhibition of 1, 1-DCE and 1, 1, 1-TCA on butane utilization. A transformation capacity term was also included in the model formation that results in cell loss due to contaminant transformation. Parameters for the model simulations were determined independently in kinetic studies with the butane-utilizing culture and through batch microcosm tests with groundwater and aquifer solids from the field test zone with the butane-utilizing culture added. In microcosm tests, the model simulated well the repetitive utilization of butane and cometabolism of 1.1, 1-TCA and 1, 1-DCE, as well as the transformation of 1, 1-DCE as it was repeatedly transformed at increased aqueous concentrations. Model simulations were then performed under the transport conditions of the field test to explore the effects of the bioaugmentation dose and the response of the system to tile biostimulation with alternating pulses of dissolved butane and oxygen in the presence of 1, 1-DCE (50 $\mu$g/L) and 1, 1, 1-TCA (250 $\mu$g/L). A uniform aquifer bioaugmentation dose of 0.5 mg/L of cells resulted in complete utilization of the butane 2-meters downgradient of the injection well within 200-hrs of bioaugmentation and butane addition. 1, 1-DCE was much more rapidly transformed than 1, 1, 1-TCA, and efficient 1, 1, 1-TCA removal occurred only after 1, 1-DCE and butane were decreased in concentration. The simulations demonstrated the strong inhibition of both 1, 1-DCE and butane on 1, 1, 1-TCA transformation, and the more rapid 1, 1-DCE transformation kinetics. Results of tile field demonstration indicated that bioaugmentation was successfully implemented; however it was difficult to maintain effective treatment for long periods of time (50 days or more). The demonstration showed that the bioaugmented experimental leg effectively transformed 1, 1-DCE and 1, 1-DCA, and was somewhat effective in transforming 1, 1, 1-TCA. The indigenous experimental leg treated in the same way as the bioaugmented leg was much less effective in treating the contaminant mixture. The best operating performance was achieved in the bioaugmented leg with about over 90%, 80%, 60 % removal for 1, 1-DCE, 1, 1-DCA, and 1, 1, 1-TCA, respectively. Molecular methods were used to track and enumerate the bioaugmented culture in the test zone. Real Time PCR analysis was used to on enumerate the bioaugmented culture. The results show higher numbers of the bioaugmented microorganisms were present in the treatment zone groundwater when the contaminants were being effective transformed. A decrease in these numbers was associated with a reduction in treatment performance. The results of the field tests indicated that although bioaugmentation can be successfully implemented, competition for the growth substrate (butane) by the indigenous microorganisms likely lead to the decrease in long-term performance.

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Studies on the Species Crossabilities in the Genus Pinus and Principal Characteristics of F1 Hybrids (일대잡종송(一代雜種松)의 교배친화력(交配親和力)과 특성(特性)에 관(關)한 연구(硏究))

  • Ahn, Kun Yong
    • Journal of Korean Society of Forest Science
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    • v.16 no.1
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    • pp.1-32
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    • 1972
  • By means of the interspecific hybridization in the Sub-genus Diploxylon of the Genus Pinus, $F_1$ hybrids of Pinus rigida${\times}$elliottii, Pinus rigida${\times}$radiata, P. rigida${\times}$serotina and P. densiflora${\times}$thunbergii had been produced. And on the basis of the crossabilities of these hybrids the taxonomic affinities of these pines were examined. And the needle characteristics of these hybrid and the occurence of phenolic substances in these $F_1$ hybrid were also investigated to see the potential usefulness of these characteristics for the diagnosis of the taxonomic affinity. And, the growth performances of the $F_1$ hybrids have also been compared with those of parental species. In order to contribute to the establishment of the hybrid seed orchard the introgression phenomena between P. densiflora and P. thunbergii in the eastern coastal area have also been investigated along with the investigation of the heterozygosity of plus trees of P. densiflora growing in the clone bank in Suwon. And the results were summarized as follows. 1. On the basis of crossabilities as well as on the taxonomic affinities according to the systems of Shaw, Pilger and Duffield, it has been proven that the parental species of those hybrids are of close affinities and range of the fertile hybrid seed production rate was as high as 28-58% in the best hybrid combination (Table 13). 2. Among those hybrids, the ${\times}$ Pinus, rigiserotina hybrid seemed to be most promising in the growth performance exhibiting 109-155% more volume growth compared to the seed parent with the statistic significance of 1% level (Tables 16 and 17). 3. Notwithstanding the fact that the all of the pollen parents are cold tender, all hybrids exhibit cold hardiness as much as their seed parent and it seems to suggest that the characteristics of cold hardiness were transmitted from the seed parent. 4. Though a striking difference in needle length was observed between the parental species of each hybrid, it was difficult to distinguish each hybrid from their seed parent by the needle length except ${\times}$P. rigiserotina which is characterized by long needle which is 65% more longer than the needle of the seed parent (Table 21). 5. With regard to the anatomical characteristics of needle, the hypoderm is apparently thicker in most of the $F_1$ hybrid pines and the characteristics of resin canals are dominated by medial in most $F_1$ hybrid. And, the fibrovascular bundles were apart as were in their seed parent. Therefore it was found to be possible to distinguish the hybrids pines from their parents by the needle characteristics. And, it is to be noticed that the ${\times}$P. densithunbergii was more close to the pollen parent having RDI value of 0.73 (Fig.l, Table 22). 6. It has been demonstrated that ${\times}$P. rigielliottii, ${\times}$P. rigiradiata and ${\times}$P. rigitaeda have a phenolic substance (No.7) of light yellow at Rf-0.46, same as their seed parent, but no trace of phenolic substance was observed in their pollen parent. This fact will serve as an important criteria for early identification of hybridity in progeny testing. However, the fact that both of ${\times}$P. rigiserotina and ${\times}$P. densithunbergii exhibit the same reactions of phenolic substances as well their parental species seems to indicate the close affinities between the parental species of the respective hybrid (Fig.2, Table 23). 7. The separation and the reaction of phenolic substance developed on TLC were found to be same in the same species showing no variations between the individuals, and no variations due to tree part of sampling, tree age or pollen sources. And the reaction was also observed regardless of the not varied by the kind of developing solvent whether it is Aceton-Chloroform (3:7 v/v) or Benzene-Methanol-Acetic acid (90:16:8 v/v). 8. The introgression phenomena of natural Pinus densifiora stand in both east and west coastal area indicates that the major part of the red pines investigated are all heterozygous and the heterozygosity of pines are higher in the west coast than in the east coast(Tables 24 and 25). 9. Based on the RDI, among the plus trees of Pinus densiflora selected in Korea and Japan as well, no pure P. densiflora has been found. Since all of the sample trees of Pinus densiflora were found to be as heterozygous bearing part of the characteristics of P. thunbergii, those red pines were considered to be natural heterotic hybrid pines(Figs. 3 and 4. Tables 26 and 27).

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Effect of Ratio of Demineralized Bone Powder with Alginate Microcapsules on Articular Cartilage Regeneration (탈미네랄 골분이 비율별로 포접된 알지네이트 미세캡슐을 이용한 조직공학적 연골재생)

  • Kim, A Ram;Kim, Hye Min;Lee, Jung Keun;Lee, Ji Hye;Song, Jeong Eun;Yoon, Kun Ho;Lee, Dongwon;Khang, Gilson
    • Polymer(Korea)
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    • v.36 no.6
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    • pp.768-775
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    • 2012
  • Alginate, obtained from the seaweeds, is a widely used biomaterial for cell transplantation, since its positive effect on viability of capsulized cells and its easier encapsulation capability of living cells. Demineralized bone powder (DBP), derived from the natural bone tissue, is widely applied for clinical trials for its low rate of reaction and antigenicity. A chondrocyte was seeded into an alginate with DBP of different contents, and a microcapsule was produced. The adhesion and proliferation of cells was observed through the MTT analysis, and the PCR was applied to estimate the content of the glycosaminoglycan (sGAG) and collagen, and confirm the specific genetic pattern of the chondrocytes. Also, the alginate microcapsule where the chondrocyte is seeded was extracted after transplantation under the skin of a nude mouse, and was immunochemically stained. The experimental result confirmed that the alginate microcapsule containing 1% of DBP not only showed the highest proliferation of cell but had a positive effect of chondrocytes by the interaction between the alginates and the growth factor in DBP. It can be expected that the microcapsule with application of the alginates and DBP might be an appropriate scaffold for tissue engineering.

Effects of Bisphenol A on the Placental Function and Reproduction in Rats (Bisphenol A가 흰쥐의 태반 기능과 출산에 미치는 영향)

  • Lee, Chae-Kwan;Kim, Seog-Hyun;Moon, Deog-Hwan;Kim, Jeong-Ho;Son, Byung-Chul;Kim, Dae-Hwan;Lee, Chang-Hee;Kim, Hwi-Dong;Kim, Jung-Won;Kim, Jong-Eun;Lee, Chae-Un
    • Journal of Preventive Medicine and Public Health
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    • v.38 no.3
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    • pp.330-336
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    • 2005
  • Objectives : The aim of this study was to investigate the effects of bisphenol A (BPA), an estrogen-like environmental endocrine disrupter, on the placental function and reproduction in rats. The mRNA levels of the placental prolactin-growth hormone(PRL-GH) gene family, placental trophoblast cell frequency and reproductive data were analyzed. Methods : The pregnancies of F344 Fisher rats ($160g{\pm}20g$) were detected by the presence of the copulatory plug or sperm in the vaginal smear, which marked Day 0 of pregnancy. Pregnant rats were divided into three groups. The control group was intraperitoneally injected with a sesame oil vehicle. The two remaining groups were injected with 50 or 500 mg/kg B.W/day of BPA, resuspended in sesame oil, on either days 7 to 11 or 16 to 20 of pregnancy, with the rats sacrificed on either day 11 or 20, respectively. The mRNA levels of PRL-GH and Pit-1a and b isotype genes were analyzed by Northern blot hybridization and reverse transcription-polymerase chain reaction. The hormone concentrations were analyzed by radioimmunoassay, and the frequency of the placental trophoblast cells observed by a histochemical study. Reproductive data, such as the placental weight and litter size, were surveyed on day 20. The fetal weight was surveyed for 4 weeks after birth. A statistical analysis was carried out using the SAS program (version 8.1). Results : The mRNA levels of the PRL-GH gene family, such as placental lactogen I, Iv and II, prolactin like protein A, C and Cv, and decidual prolactin-related protein were significantly reduced due to BPA exposure. The mRNA levels of the Pit-1a and b isotype genes, which induce the expression of the PRL-GH gene family in the rat placenta, were also reduced due to BPA exposure. The PL-Iv and PL-II concentrations were reduced in the BPA exposed group. During the middle to last stage of pregnancy (Days 11-20), a high dose of BPA exposure reduced the frequency of spongiotrophoblast cells, which are responsible for the secretion of the PRL-GH hormones. Reproductive data, such as the placental and fetal weights and the litter size, were reduced, but that of the pregnancy period was extended in the BPA exposed compared to the control group. Conclusions : BPA disrupts the placental functions in rats, which leads to reproductive disorders.

Toll-like Receptor 4-mediated Apoptotic Cell Death in Primary Isolated Human Cervical Cancers (부인과질환 특이적 종양의 TLR4 매개성 apoptosis 유발에 관한 연구)

  • Won, Jinyoung;Hong, Yunkyung;Park, Sookyoung;Kim, Joo-Heon;Hong, Yonggeun
    • Journal of Life Science
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    • v.28 no.6
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    • pp.718-725
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    • 2018
  • Toll-like receptor 4 (TLR4) has been implicated in cell proliferation and apoptosis in several types of cancer. In this study, the impact of TLR4 activation on apoptotic cell death in gynecologic cancers induced by lipopolysaccharide (LPS) was investigated. Cervical cancer cell lines were produced from isolated surgical specimens supplied by Paik Hospital. The primary cultures of normal myometrium and gynecologic cancers, including cervical, endometrial, and ovarian cancers, were used to examine the differences in morphological characteristics between normal and cancerous cells. A reverse transcription polymerase chain reaction analysis was used to determine the relative expression levels of TLR4 gene involved in apoptosis-associated signaling in cervical cancer cells. The cancer cell colonies showed a tendency to reach high levels of confluency compared with normal cells. In addition, an enhanced growth rate and loss of contact inhibition were observed in gynecologic cancer cells compared with normal cells (doubling times of 16.6 hr vs. 26 hr, respectively). The expression level of ITGA5, an alpha-5 integrin marker, was upregulated in normal myometrial cells, but this tendency was not exhibited in cervical cancer cells. Furthermore, p53 tumor suppressor gene expression was upregulated, whereas TLR4 and caspase-3 gene expressions were downregulated in cervical cancer cells. Notably, the expression levels of TLR4 and caspase-3 were increased significantly in LPS-treated cancer cells compared with those in non-LPS-treated cells. These results suggest that the TLR4-mediated caspase-dependent apoptotic signaling pathway could be suggested as a therapeutic target for the treatment of gynecologic cancers, including cervical cancers.

Establishment of a Radiation-Induced Fibrosis Model in BALB/c Mice (BALB/c 마우스를 이용한 방사선섬유증 모델 확립)

  • Ryu, Seung-Hee;Lee, Sang-Wook;Moon, Soo-Young;Oh, Jeong-Yoon;Yang, Youn-Joo;Park, Jin-Hong
    • Radiation Oncology Journal
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    • v.28 no.1
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    • pp.32-38
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    • 2010
  • Purpose: Although radiation-induced fibrosis is one of the common sequelae occurring after irradiation of skin and soft tissues, the treatment methods are not well standardized. This study aimed to establish the skin fibrosis mouse model by fractionated radiation for the further mechanism studies or testing the efficacy of therapeutic candidates. Materials and Methods: The right hind limbs of BALB/c mice received two fractions of 20 Gy using a therapeutic linear accelerator. Early skin damages were scored and tissue fibrosis was assessed by the measurement of a leg extension. Morphological changes were assessed by H&E staining and by Masson's Trichrome staining. TGF-${\beta}1$ expression from soft tissues was also detected by immunohistochemistry and PCR. Results: Two fractions of 20 Gy irradiation were demonstrated as being enough to induce early skin damage effects such as erythema, mild skin dryness, dry and wet desquamation within several weeks of radiation. After 13 weeks of irradiation, the average radiation-induced leg contraction was $11.1{\pm}6.2mm$. Morphologic changes in irradiated skin biopsies exhibited disorganized collagen and extracellular matrix fibers, as well as the accumulation of myofibroblasts compared to the non-irradiated skin. Moreover, TGF-${\beta}1$ expression in tissue was increased by radiation. Conclusion: These results show that two fractions of 20 Gy irradiation can induce skin fibrosis in BALB/c mice accompanied by other common characteristics of skin damages. This animal model can be a useful tool for studying skin fibrosis induced by radiation.

Effect of Feeding Plum and Red Ginseng Marc on Vital Reaction in Broiler Stress (매실과 홍삼 부산물이 스트레스에 대한 육계 생체 반응에 미치는 영향)

  • Bong, Mi-Hee;Ji, Sang-Yoon;Park, Jun-Cheol;Moon, Hong-Kil;Lee, Sang-Cheul;Lee, Jun-Heon;Hong, Joon-Ki
    • Korean Journal of Poultry Science
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    • v.38 no.3
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    • pp.213-223
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    • 2011
  • This study was conducted to test the efficacy of plum (Prunus mume) and red ginseng (Panax ginseng C.A Meyer) marc as stress inhibitors under heat stress and lipopolysaccharide (LPS) challenge in broilers by investigating their effects on blood biochemical parameters, immunoglobulin concentration and splenic cytokine mRNA expressions. A total of one hundred ninety-two 1-d-old male broiler chicks (Ross 308) were divided into 2 stress conditions (heat and LPS) experiments. Each experiment was divided into 4 treatment groups with 8 replicates of 3 birds in each group. NC (negative control, no immune substances), PC (positive control, 25 ppm ${\beta}$-glucan), PM (1% plum marc) and RGM (3% red ginseng marc) treatments were administered with respective substance through water supplementation. During heat stress, The Ca/Mg ratio in PM and RGM was significantly decreased in comparison with that of NC (P<0.05). The immunoglobulin M was significantly lower in PM than in NC (P<0.05). Expression patterns of splenic cytokine mRNAs (IL-1, IL-2 and IL-6) were similar over the treatment. Expression rates of IL-1 and IL-2 in PM were significantly decreased in comparison with NC. Also, expression rates of IL-1, IL-2 and IL-6 were significantly lower in RGM than in NC (P<0.05). In conclusion, the dietary supplementation of plum and red ginseng marc improved coping ability to heat stress by preventing Ca/Mg ratio increment and by inhibiting inflammatory response in broiler chicks. However, it is necessary to determine optimal dietary level of red ginseng marc for improving growth performances in broiler chickens. These results suggest the possibility that plum and red ginseng marc could be used as the stress inhibitor under heat stress and inflammatory response in broiler chicks.

Clinical and Bacteriologic Efficacy of Cefdinir on Pharyngitis and Pharyngotonsillitis Caused by Group A Beta Hemolytic Streptococci in Children (Group A-beta Hemolytic Streptococci에 의한 소아 인두편도염에 있어서 Cefdinir의 항균력과 임상적 및 세균학적 효과에 관한 연구)

  • Chung, Ji-Young;Sin, Seon-Hee;Ahn, Young-Min;Ahn, Byung-Moon;Sin, Young-Gyu;Bae, Young-Min;Park, Soo-Eun;Kim, Jong-Guk;Lee, Jong-Guk;Ma, Saung-Hyuk;Chang, Jin-Kun;Cha, Sung-Ho
    • Pediatric Infection and Vaccine
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    • v.10 no.1
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    • pp.95-101
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    • 2003
  • Objective : To determine the clinical and bacteriologic efficacy and safety of Cefdinir in acute pharyngitis and pharyngotonsillitis caused by group A beta hemolytic streptococci in pediatric patients. Methods : Children aged 3 through 12 years who visited the hospitals enrolled in this study with signs and symptoms of pharyngitis or pharyngotonsillitis since May to December 2002, were taken throat culture and given Cefdinir(12 mg/kg/day, in three divided doses) for 7 days. Two hundred thirty five patients were enrolled and 90 patients who showed positive culture results were followed up for the signs and symptoms during the treatment to determine clinical efficacy. Follow-up culture were done at the end of the study and bacteriologic efficacy was determined Results : Ninety out of 235 patients who visited the hospitals with the signs and symptoms of pharyngitis showed positive growth on throat culture. Seventy nine patients were clinically and microbiologically assessable. The bacteriologic eradication rates of S. pyogenes were 100% in all the children treated with Cefdinir. Clinical cure rates were not different between less than 7 days-treated group(75%) and just 7 days-treated group(98.6%)(P=0.077). Two patients reported adverse reaction during Cefdinir treatment. Antimicrobial sensitivity of Cefdinir against S. pyogenes was 100% with range of MIC being less than 0.5 ${\mu}g/mL$. Conclusion : It seems that Cefdinir is one of reliable and well-tolerated drugs for the treatment of group A beta hemolytic streptococcal pharyngotonsillitis in children. It needs to be investigated short term efficacy in terms of improving drug compliance and impact of economic point of view. We can recognized that still high rate of the erythromycin resistant group A streptococci in our community should be monitored every year.

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Screening of Resistant Genetic Resources to Stink Bug in Soybean (톱다리개미허리노린재 저항성 콩 유전자원 탐색)

  • Oh, Sea-Kwan;Baek, In-Yeol;Hwang, Pil-Seong;Kang, Sung-Tag;Suh, Deug-Yong;Park, Geum-Yong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.52 no.2
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    • pp.140-145
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    • 2007
  • This study was carried out to develop effective test method by soybean stink bug and to screen resistant genetic resources against soybean stink bug. The damage pod rate by stink bug showed 40% of most soybean varieties and was selected about 10% low of 10 varieties by 298 variety and degree in soybean at first year Stink bug damage rate research for 102 varieties that stink bug damage rate lowed at first year showed 10% low of 12 varieties and from 20% to 30% of the other varieties. So testing material is "Ilpumgeumjeongkong" to develop for effective test method soybean stink bug and result for stink bug damage rate research of according to growth stage showed rapidly high more full seed than full pod. Full seed stage (R6) was highest to 35.5% for stink bug damage rate. Result of resistant genetic resources selection according to stink bug damage pod rate was lowed of best to 10.3% for "Peking, Sorogkong, Hwangsaegjunjeari and Sobaeknamulkong" in the order. Also, stink bug damage seed rate was similar too. So "Peking, Sorogkong, Hwangsaegjunjeari and Sobaeknamulkong" were thought resistant variety against stink bug. Additional study carried out with "Peking and Sorogkong" so that concretely investigate about stink bug's refuse reaction. This result showed 10.0% for Peking and 14.2% for Sorogkong at R6 stage. But, damage pod rate was rapidly lowed.

Expression of the Circadian Clock Genes in the Mouse Gonad (생쥐 생식소의 발달 단계에 따른 일주기성 유전자 발현에 관한 연구)

  • Chung Mi-Kyung;Choi Yoon-Jeong;Jung Kyenng-Hwa;Kim Eun-Ah;Chung Hyung-Min;Lee Sook-Hwan;Yoon Tae-Ki;Chai Young-Gyu
    • Development and Reproduction
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    • v.8 no.1
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    • pp.57-64
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    • 2004
  • This study was carried out to examine the expression of the circadian clock genes in the mouse ovary and testis at different developmental stages. Expression of Period1(Per 1), Period2(Per2), Period3(Per3), Cryptochrome1(Cry1), Cyptochrome2(Cry2), Clock Small and Prokineticin1 and Prokineticin2 receptor(Prok1r, Prok2r) genes in mouse ovary was explored by semiquantitative reverse transcription Polymerase chain reaction(RT-PCR) according to the developmental stage(post partum day; ppd 1, 7, 10, 21 and 35). Immunohistochemistry using PER1 antibody was also analyzed. The differential expression pattern of clock genes was presented according to stages of the mouse ovarian development (ppd 1, 7, 10, 21 and 35). In the cases of ovaries, at the starting point of follicle growth at ppd 7 and 10, the clock gene expression patterns were changed vastly. According to the developmental stages, the clock genes were highly expressed at ppd 7 and 10 in mouse testis also. Receptors for Prok2, the circadian output molecule of SCN, were also expressed in ovary at ppd 7 and in testis at ppd 1 and 7, respectively. Immnunohistochemical analysis of PER1 showed positive signals in the cytoplasm of oocytes and granulosa cells. The level or PER1 expression was increased in cells at the spermatogonia and the condensing spermatids. The expression pattern of Perl and localization of PER1 were showed similar patterns according to the developmental stages in ovary and testis. Taken together, it could be observed that the expression of clock genes was highly correlated with gonadal development and germ cell differentiation in mice. Therefore, in this study, circadian programming of the genes in the ovary and testis is strongly imposed across a wide range of core reproductive cycles and normal development of gametes. Although the existence of circadian genes is clearly investigated, further studies on the direct evidence is required for the understanding of the relationship between circadian genes and regulation of gonadal differentiation and germ cell development.

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