• Title/Summary/Keyword: Strain Control

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Culture Conditions of Aspergillus oryzae in Dried Food-Waste and the Effects of Feeding the AO Ferments on Nutrients Availability in Chickens (건조한 남은 음식물을 이용한 Aspergillus oryzae균주 배양조건과 그 배양물 급여가 닭의 영양소 이용률에 미치는 영향)

  • Hwangbo J.;Hong E. C.;Lee B. S.;Bae H. D.;Kim W.;Nho W. G.;Kim J. H.;Kim I. H.
    • Korean Journal of Poultry Science
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    • v.32 no.4
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    • pp.291-300
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    • 2005
  • Two experiments were carried out to assess the appropriate incubation conditions namely; duration, moisture content and the ideal microbial inoculant for fermented dried food waste(EW) offered to broilers. The nutrient utilization of birds fed the FW diets at varying dietary inclusion rates was also compared with a control diet. In Experiment 1, different moisture contents(MC) of 30, 40, 50 and $60\%$ respectively were predetermined to establish the ideal duration of incubation and the microbial inoculant. A 1mL Aspergillus oryzae(AO) $(1.33\times10^5\;CFU/mL)$ was used as the seed inoculant in FW. This results indicated that the ideal MC for incubation was $40\~50\%$ while the normal incubation time was > 72 hours. Consequently, AO seeds at 0.25, 0.50, 0.75 and 1.00mL were inoculated in FW to determine its effect on AO count. The comparative AO count of FW incubated for 12 and 96 hours, respectively showed no significant differences among varying inoculant dosage rates. The FW inoculated with lower AO seeds at 0.10, 0.05 and 0.01mL were likewise incubated for 72 and 96 hours, respectively and no changes in AO count was detected(p<0.05). The above findings indicated that the incubation requirements for FW should be $%40\~50\%$ for 72 hours with an AO seed incoulant dosage rate of 0.10mL. Consequently, in Experiment II, after determining the appropriate processing condition for the FW, 20 five-week old male Hubbard strain were used in a digestibility experiment. The birds were divided into 4 groups with 5 pens(1 bird per pen). The dietary treatments were; Treatment 1 : Control(Basal diet), Treatment 2 : $60\%$ Basal+4$40\%$ FW, Treatment 3 : $60\%$ $Basal+20\%\;FW+20\%$ AFW(Aspergillus oryzae inoculate dried food-waste diet) and Treatment 4: $60\%$ Basal+$40\%$ Am. Digestibility of treatment 2 was lowed on common nutrients and amino acids compared with control(p<0.05) and on crude fat and phosphorus compared with AFW treatments(T3, T4)(plt;0.05). Digestibility of treatment 3 and 4 increased on crude fiber and crude ash compared treatment 2 (p<0.05). Digestibility of control was high on agrinine, leucine, and phenylalnine of essential amino acids compared with treatment 3 and 4(p<0.05), and diestibility of treatment 3 and 4 was improved on arginine, lysine, and threonine of essential amino acids. Finally, despite comparable nutrient utilization among treatments, birds fed the dietary treatment containing AO tended to superior nutrient digestion to those fed the $60\%$ Basa1+$40\%$ FW.

The Difference in Chemokine Expression in Airway Epithelial Cells According to the Virulence of Tubercle Bacilli (결핵균 독성 여부에 따른 기도 상피세포의 Chemokine 발현에 관한 연구)

  • Kwon, O-Jung;Kim, Ho-Joong;Kim, Jung-Hee;Kim, Ho-Cheol;Suh, Gee-Young;Park, Jeong-Woong;Park, Sang-Joon;Chung, Man-Pyo;Choi, Dong-Chull;Rhee, Chong-H.
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.4
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    • pp.729-741
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    • 1997
  • Background : We have recently reported that airway epithelial cells can produce RANTES and IL-8 in response to the stimulation of tubercle bacilli suggesting a certain role of airway epithelial cells in the pathogenesis of pulmonary tuberculosis. The pathogenesis of tuberculosis is determined by several factors including phagocytosis, immunological response of host, and virulence of tubercle bacilli. Interestingly, there have been reports suggesting that difference in immunological response of host according to the virulence of tubercle bacilli may be related with the pathogenesis of tuberculosis. We, therefore, studied the expressions and productions of RANTES and IL-8 in airway epithelial cells in response to tubercle bacilli(H37Rv, virulent strain and H37Ra, avirulent strain), in order to elucidate the possible pathophysiology of pulmonary tuberculosis. Methods : Peripheral blood monocytes were isolated from normal volunteers. Peripheral blood monocytes (PBM) were stimulated with LPS($10{\mu}g/ml$), H37Rv, or H37Ra($5{\times}10^5$ bacilli/well) along with normal control for 24 hours. A549 cells were stimulated with supernatants of cultured PBM for 24 hours. ELISA kit was used for the measurement of $TNF{\alpha}$ and IL-$1{\beta}$ production in supernatants of cultured PBM and for the measurement of RANTES and IL-8 in supernatants of cultured A549 cells. Northern blot analysis was used for the measurement of RANTES and IL-8 mRNA expression in cultured A549 cells. Results : $TNF{\alpha}$ and IL-$1{\beta}$ productions were increased in cultured PBM stimulated with LPS or tubercle bacilli(H37Rv or H37Ra) compared with the control. There was, however, no difference in $TNF{\alpha}$ and IL-$1{\beta}$ production between cultured PBM stimulated with H37Rv and H37Ra. RANTES and IL-8 expressions and productions were also increased in cultured A549 cells stimulated with LPS or tubercle bacilli compared with the control. RANTES and IL-8 mRNA expressions were significantly increased in cultured A549 cells stimulated with H37Ra-conditioned media(CM) compared with A549 cells stimulated with H37Rv-CM (p<0.05). However, there was no difference in RANTES and IL-8 productions between A549 cells stimulated with H37Rv-CM and H37Ra-CM. Conclusion : Airway epithelial cells can produce the potent chemokines such as RANTES and IL-8, in response to the stimulation of tubercle bacilli. These results suggest that airway epithelial cells may play a certain role in the pathogenesis of pulmonary tuberculosis. However, the role of airway epithelial cells in the pathogenesis of tuberculosis according to the virulence of tubercle bacilli was not clear in this study.

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Production of Concentrated Blueberry Vinegar Using Blueberry Juice and Its Antioxidant and Antimicrobial Activities (블루베리 농축식초 제조 및 이들의 항산화 및 항균 활성)

  • Oh, Hyeonhwa;Jang, Sowon;Jun, Hyun-Il;Jeong, Do-Youn;Kim, Young-Soo;Song, Geun-Seoup
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.6
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    • pp.695-702
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    • 2017
  • This study was carried out to investigate the effects of seed vinegar on antioxidant activity and antimicrobial activities of concentrated blueberry vinegar (CBV). Of the nine strains of yeast and six strains of acetic acid bacteria provided by the Microbial Institute for Fermentation Industry, each strain of yeast (Saccharomyces cerevisiae SRCM 100610, showing the highest ethanol content) and acetic acid bacteria (Acetobacter pasteurianus SRCM 101342, showing the highest total acidity) was selected for production of CBVs. Sugar content, pH, total acidity, total phenolic content (TPC), and browning intensity (280 nm and 420 nm) in CBVs using concentrated blueberry juice were $11.05{\sim}12.70^{\circ}Brix$, 2.63~2.98, 1.65~5.72%, 3.03~4.24 mg/mL, 0.95~1.50, and 0.11~0.20, respectively. Sugar content and total acidity of CBVs increased upon addition of seed vinegar, whereas pH, TPC, and browning intensity decreased. Of all CBVs with various additions of seed vinegar, the control showed the lowest $EC_{50}$ values in DPPH radical scavenging assay, ABTS radical scavenging assay, and reducing power (23.80, 19.48, and 79.21 dilution factor, respectively), whereas the 40% seed vinegar group showed the highest clear zone diameter values for Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, and Bacillus cereus (4.31, 4.59, 5.81, and 3.97, respectively). Antioxidant activities of CBVs were closely correlated with their TPC, browning intensity at 280 nm, pH, and total acidity values, showing correlation determination coefficient ($R^2$) values higher 0.82. However, antimicrobial activities of CBVs were closely correlated with their pH and total acidity values, showing higher $R^2$ values more than 0.92. These results suggest that CBVs using concentrated blueberry juice, S. cerevisiae SRCM 100610, and A. pasteurianus SRCM 101342 may be useful as potentially functional foods for enhancing health.

Outbreak of Shigellosis Occurred in a Preschool and Two Elemetary Schools in Mapo-Gu, Seoul (서울시 마포구 관내 어린이집 및 초등학교에서 집단 발병한 세균성 이질)

  • Park, Tae Su;Lee, Ho Jun;Kim, Su Yeon;Lee, Dong Woo;Kim, Jae Yoon;Baik, Ji Na;Park, Yu Mi;Park, Mi Sun;Lee, Bok Kwon
    • Pediatric Infection and Vaccine
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    • v.13 no.1
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    • pp.71-77
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    • 2006
  • Purpose : Shigellosis is still an important cause of acute food-borne diarrheal diseases throughout the world. We investigated the transmission routes and clinical course through the outbreak of shigellosis in Mapo-Gu, Seoul. Methods : From October 7th to November 19th in 2005, Mapo-Gu public health center had referred 32 patients with confirmed or suspected shigellosis to the National Medical Center. We had investigated source of infection, epidemiology, laboratory findings, and clinical course of the cases occurred during this outbreak. Results : Among 32 patients, 24 patients had been confirmed with shigellosis, 8 patients had been diagnosed with suspected shigellosis. They ranged in age from 5 months to 12 years old and their mean age was 6.5 years. The clinical manifestations were as follows; diarrhea, fever, abdominal pain and asymptomatic condition. Symptoms had sustained for 3.7 days on the average. S. sonnei were cultured by rectal swab and founded to be resistant to ampicillin and TMP/SMX except to 3rd generation cephalosporin. After treatment with antibiotics such as cefixime and ceftriaxone or imipenem and conservative treatment with electrolyte and fluid replacement for 5~7 days, Stool cultures of the rectal swab grew no Shigella in these cases except 3 cases. Conclusion : An outbreak of shigellosis had occurred in a preschool and elementary school children. From the same results of antimicrobial susceptibility and pulsed-field gel electrophoresis patterns in this study, we suggest that the outbreak of shigellosis in this report had been originated from a single strain. According to all negative results about suspected food and water cultures, we couldn't find out source of infection. Through materials offerred by Mapo-Gu Public Health Center, we presumed the trasmission routes probably were person-to-person.

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Effects of Dietary Addition of Wormwood (Artemisia sp.) on the Growth Performance, Nutrients Utilization, and Abdominal Fat Deposition of Broiler Chickens (사료내 쑥 첨가가 육계의 성장률, 영양소 이용률, 복강내 지방에 미치는 영향)

  • Lee, Sung-Jin;Song, Ki-Duk;Ko, Yung-Du;Kim, Chang-Hyun
    • Korean Journal of Organic Agriculture
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    • v.18 no.3
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    • pp.347-361
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    • 2010
  • This study was carried out to investigate the effects of wormwood (Artemisia sp.) addition on the growth performance, nutrients utilization and abdominal fat deposition of broiler chickens. Two hundred twenty five and two-day old Arboracre strain male commercial broiler chicks were distributed to 5 treatments with wormwood supplementation levels; C(O%), $T_1$ (1%), $T_2$ (3%), $T_3$ (5%), and $T_4$ (10%) and with 3 replications each with 5 birds for five weeks. Body weight gain during the experiment was improved in $T_1$ (1723.0g) compared with that of $T_3$ (1557.7g) and $T_4$ (1450.7g) (P<0.05). Feed intake was significantly (P<0.05) increased as the levels of wormwood addition increased (C: 2653.8g, T1: 2852.0, $T_2$: 2900.3, $T_3$: 2900.7g, T4: 2954.7g). Feed conversion rate (feed/gain) was significantly (P<0.05) increased as the levels of wormwood addition increased (C: 1.55, $T_1$: 1.66, $T_2$: 1.70, $T_3$: 1.86, $T_4$: 2.04). The days reaching to 2.0kg of body weight were expected to be 43.2 days in control, whereas those of group T1 were reduced to 42.6 days by 0.6 day from control. Nutrient utilization and abdominal fat deposition in the experiment were significantly decreased (P<0.05) and small intestine contents in the broilers was significantly increased (P<0.05) as the levels of wormwood addition increased. Therefore, although there was no significant improvement for the performance of broiler chickens with the dietary supplementation of wormwood meal, less than 1% addition of wormwood to broiler diets might have beneficial for human health by reducing the abdominal fat deposition of the broiler chicknens.

Direct Action of Genistein on the Hypothalamic Neuronal Circuits in Prepubertal Female Rats : Estrogen Receptor Beta($ER{\beta}$) Pathway? (미성숙한 암컷 흰쥐 시상하부의 신경회로에 미치는 Genistein의 직접 작용 : 에스트로겐 수용체 베타아형 경로?)

  • Heo, Hyun-Jin;Lee, Sung-Ho
    • Development and Reproduction
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    • v.15 no.2
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    • pp.179-185
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    • 2011
  • Some phytoestrogens in soy and red wine, for example, might have beneficiary rather than adverse effects. In particular, dietary soy intake seems to be highly correlated with protection of breast cancer, osteoporosis and cardiovascular disorders. However, questions persist on the potential adverse effects of the main soy constituent genistein (GS) on female reproductive physiology. Previously we found that prepubertal exposure to GS could activate the reproductive system of immature female rats leading to precocious puberty onset, and intracerebroventricularly (ICV) injected GS could directly activate hypothalamic kisspeptin-GnRH neuronal circuits in adult female rats. The present study was performed to examine the hypothalamus-specific GS effects in prepubertal female rats and which subtype of estrogen receptor is mediated in this GS effect. Prepubertal female rats (PND 30) were anaesthetized, treated with single dose of GS (3.4 ${\mu}g$/animal), and sacrificed at 2 hrs post-injection. To determine the transcriptional changes of reproductive hormone-related genes in hypothalamus, total RNAs were extracted and applied to the semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). ICV infusion of GS significantly lowered the transcriptional activities of mTOR (1:$0.361{\pm}0.058$ AU, p<0.001) but increased that of GAD67 (1:$1.285{\pm}0.099$ AU, p<0.05), which are known to act as an upstream modulator of kisspeptin and GnRH neuronal activities in the hypothalamus, respectively. GS administration enhanced significantly the mRNA levels of KiSS-1(1:$1.458{\pm}0.078$ AU, p<0.001), and exerted no effect on the mRNA level of kisspeptin receptor GPR-54 (1:$1.29{\pm}0.08$ AU). GnRH gene expression was significantly decreased in GS-treated group compared to control group (1:$0.379{\pm}0.196$ AU, p<0.05). There was no difference in the mRNA level of $ER{\alpha}$ in the GS-treated group compare to control group (1:$1.180{\pm}0.390$ AU, Fig. 3A). However, icv infusion of GS significantly increased the transcriptional activities of $ER{\beta}$ (1:$4.209{\pm}0.796$ AU, p<0.01, Fig. 3B). Taken together, the present study indicated that the acute exposure to GS could directly alter the hypothalamic GnRH modulating system in prepubertal female rats. Our study strongly suggested the involvement of $ER{\beta}$ pathway in GS's hypothalamus-specific action, and this idea is consistent with the GS's well-known $ER{\beta}$-mediated protective action in breast cancer.

Studies on the Effect of Fiber Reinforcing upon Mechanical Properties of Concrete and Crack Mode of Reinforoed Concrete (섬유보강이 콘크리트의 역학적 특성과 철근콘크리트의 균열성상에 미치는 영향에 관한 연구)

  • 박승범
    • Magazine of the Korean Society of Agricultural Engineers
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    • v.20 no.2
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    • pp.4645-4687
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    • 1978
  • This study was attempted to investigate the mechanical properties of concrete and crack control effects of reinforced concrete with steel and glass fiber. The experimental program includes tests on the properties of fresh concrete containing fibers, compressive strength, tensile strength, flexural strength, Young's modulus, Shrinkage and deformation of steel or glass fiber reinforced concrete. Also this study was carried out to investigate the effect of steel or glass fiber to retard the development in reinforced concrete subject to uniaxial tension and thus facilitate the use of steels of higher strength. The major conclusions that can be drawn from the studies are as follows: 1. The effect of the fibers in various mixes on fresh concrete confirmed that fibers do have a significant effect on the properties of fresh concrete, bringing much more stable and exhibiting a signiflcant reduction in surface bleeding, and that the cohesion is greatly improved and the internal resistance increases with fiber concentration. But the addition of an excess contents and length of fibers brings about the reduction of workability. 2. With the addition of steel fibers(1.5% Vol.) to concrete, the compressive strength as compared with plain concrete showed a very slight increase, but excess addition, over 1.5% Vol. of steel and glass fiber reduced its strength. 3. Splitting tensile strength of fiber reinforced concrete showed a significant increase tendency, as compared with plain concrete. In case of containing steel fiber (2.5%, 30mm), it showed that the maximum increase rate of 1.48 times as much rate, and in case of containing glass fiber (2.5%, 30mm), the increase rate of strength was 1.25 times as much rate. 4. Flexural strength of fiber reinforced concrete showed a significant tendency, as compared with plain concrete. Containing steel fiber (2.5%, 30mm) showed the maximum increase rate of 1.64 times as much rate and containing glass fiber (2.5%, 30mm) showed the increase rate of strength of 1.32 times as much rate, and in general, the 30mm length brougth the best results. 5. The strength ratio ($\sigma$b/$\sigma$c and $\sigma$t/$\sigma$c) increased, when steel fiber's average spacing was up to 3.05mm, but decreased when beyond 3.05mm, and it was confirmed that tensile or flexural strengths of steel fiber reinforced concrete are apparently governed by fiber's average spacing. 6. The compressive strain of fiber reinforced concrete showed a significant increasing tendency as the fiber was added, but Young's modulus. with the addition of steel and glass fibers, showed a slight decrease tendency. And according to the increase of flexural strength, a considerable increase was seen in toughness. 7. With the addition of fiber's the shrinkage of concrete was significantly decreased, in both case of adding steel fibers 12.5%, 30mm, and showed a significant decrease ratio, in average 30.4% and 36.7%, as compared with plain concrete. 8. With the increase of fiber volume fraction and length, the gained stress in reinforcing bar in concrete specimens increased in all crack widths, but at different rates, with the decrease of fiber diameter, the stress showed a considerable increasing tendency. And the duoform steel fibers showed the greatest improvement, as compared with the other types tested. 9. The influence of fiber dimensions in order of significanse on the machanical properties of concrete and the crack control of reinforced concrete was explained as follows: content, length, aspect ratio and dimeter.

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Diagnosis of Irrigation Time Based on Microchange of Stem Diameter in Greenhouse Tomato (온실재배 토마토의 농직경 변화에 의한 관개시기 진단)

  • 이변우
    • Journal of Bio-Environment Control
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    • v.6 no.4
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    • pp.250-257
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    • 1997
  • Stem diameter and shoot fresh weight of tomato grown in greenhouse were measured non-destructively at 10 minutes interval from 1 to 16 July, 1996 with displacement detector using strain gauges and with suspension-type load cell, respectively, and simultaneously were measured soil water potential, transpiration and solar radiation. Ample water was irrigated before experiment, and thereafter, irrigations were made on the next morning when visual symptoms of wilting appeared. Shoot fresh weight and stem diameter showed very similar patterns in diurnal changes which are characterized by predawn maximum and afternoon minimum and in long- term evolutions, suggesting that stem diameter shrinkage and expansion are closely related to plant water content and growth, respectively, Shoot weight and stem diameter reached minimum values a little later than the time on which transpiration showed maximum. The daily net gains of fresh weight(DG) and stem diameter(DI) showed significantly Positive correlations with solar radiation in those days on which plants were not water-stressed. However, Dl and DG on those days of water stress showed much lower values than expected from the relationships between solar radiation and them. Transpiration was much lower than the expected potential transpiration on 10 July, implying that plants were water-stressed. In this case water stress was not detected from visual symptom of wilting and/or soil water potential, but was able to be identified by the lower DI and DG than the expected. The maximum contraction of stem diameter(MC) and the maximum loss of fresh weight(ML) during daytime showed significantly positive correlations with solar radiation in those days on which plants were not water-stressed and were observed greater than expected from the relationships on severely water-stressed days. But mild water stress could not be discernable by ML and MC. It would be concluded that the daily net gains of fresh weight and/or stem diameter could be used as criteria for diagnosing the water status of tomato and for triggoring the onset of irrigation in automatic system.

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Effect of Recombinant CagL Immunization on the Gastric Diseases Induced by Helicobacter pylori in Mongolian gerbils (CagL 재조합 단백질 접종후에 Mongolian gerbil에서 나타나는 Helicobacter pylori 감염에 대한 반응)

  • Bak, Eun-Jung;Jang, Sung-Il;Choi, Yun-Hui;Kim, Jin-Moon;Kim, Ae-Ryun;Kim, Ji-Hye;Woo, Gye-Hyeong;Yoo, Yun-Jung;Lee, Sung-Haeng;Cha, Jeong-Heon
    • Korean Journal of Microbiology
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    • v.48 no.2
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    • pp.109-115
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    • 2012
  • Helicobacter pylori is an important factor of chronic gastritis, digestive ulcer, and stomach cancer. CagL, a virulence factor of H. pylori, is well-known as a pilus protein which acts as adhesion to host cell and a component of Type 4 secretion system. In this study, we evaluated the protective response of recombinant CagL protein (rCagL) using Mongolian gerbil animal model for H. pylori infection. The cagL gene was cloned from 26695 H. pylori followed by over-expression and purification of the protein in E. coli. Mongolian gerbils were immunized with rCagL protein mixed with aluminum adjuvant via intramuscular injections once a week during 4 weeks. At a week after the last immunization, the Mongolian gerbils were administrated with H. pylori 7.13 strain into the stomach and sacrificed to measure antibody titer on rCagL by ELISA and bacterial colonization in the stomach, and to examine the histopathological changes and cytokine expression at 6 week after challenge. Antibody titers on recombinant protein were significantly increased from a week after the first immunization. There was no significant change of the number of bacterial colony between control group and immunized group. The relative stomach weight was significantly decreased in immunized group, but the significant change of histopathological assessment was not observed in the stomach. Cytokine expression such as IL-$1{\beta}$ and KC also was not significantly different between control and immunized groups. These results indicate that rCagL could effectively induce the formation of the specific IgG antibodies. However, bacterial colonization and histopathological lesions could not be inhibited by the immunization in the stomach, indicating not enough protection against H. pylori infection. We consider that along with CagL other adequate antigens could be needed stimulating immune response and inducing protective effects against gastric disease, and also a better adjuvant could be considered.

Plant Growth Promotion of Calystegia soldanella and Ischaemum anthephoroides by the Strain Penicillium citrinum KACC43900 (Penicillium citrinum KACC43900에 의한 갯메꽃과 갯쇠보리의 생장촉진활성)

  • You, Young-Hyun;Hwang, Jeong-Sook;Yoon, Hyeok-Jun;Khan, Sumera Afzal;Rim, Soon-Ok;Bae, Jeong-Jin;Choo, Yeon-Sik;Lee, In-Jung;Kong, Won-Sik;Lee, Byung-Moo;Kim, Jong-Guk
    • Journal of Life Science
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    • v.20 no.9
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    • pp.1373-1377
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    • 2010
  • Coastal sand-dune plants can survive very effectively with the help of various microbes, especially ecto- and endomycorrihizae. Penicillium citrinum KACC43900 was screened according to growth promoting activity on sand-dune plants Calystegia soldanella and Ischaemum anthephoroides. In this study, coastal sand-dune plants were treated with a fungal culture filtrate of endophytic fungus P. citrinum KACC43900 to confirm the promotion of plant growth. C. soldanella and I. anthephoroides were used as representative coastal sand-dune plants, and their shoot length, plant length, and dry weight were analyzed. The shoot lengths of control C. soldanella and I. anthephoroides were 8.60 cm and 8.12 cm, and those of samples treated with fungal culture filtrates of C. soldanella and I. anthephoroides for 20 days were 16.30 cm and 10.56 cm, respectively. The plant lengths of control C. soldanella and I. anthephoroides were 14.90 cm and 14.80 cm, and those of samples treated with fungal culture filtrates of C. soldanella and I. anthephoroides for 20 days were 24.06 cm and 17.06 cm, respectively. The dry weight of C. soldanella and I. anthephoroides treated with fungal culture filtrates of C. soldanella and I. anthephoroides for 20 days were 0.163 g and 0.032 g, respectively. It was shown that the growth of shoots in C. soldanella and I. anthephoroides was promoted, 89.54% and 29.60%, by culture filtrate of P. citrinum KACC43900.