It were reported that antifungal mechanism of Enterobacter cloacae is a volatile ammonia that produced by the strain in soil, and the production of ammonia is related to the bacterial urease activity. A powerful bacterium SH14 against soil-borne pathogen Fusarium solani, which cause root rot of many important crops, was selected from a ginseng pathogen suppressive soil. The strain SH14 was identified as Bacillus subtilis by cultural, biochemical, morphological method, and $API^{circledR}$ test. From several in vitro tests, the antifungal substance that is produced from B. subtilis SH14 was revealed as heat-stable and low-molecular weight antibiotic substance. In order to construct the multifunctional biocontrol agent, the urease gene of Bacillus pasteurii which can produce pathogenes-suppressive ammonia transferred into antifungal bacterium. First, a partial BamH I digestion fragment of plasmid pBU11 containing the alkalophilic B. pasteurii l1859 urease gene was inserted into the BamH I site of pEB203 and expressed in Escherichia coli JM109. The recombinant plasmid was designated as pGU366. The plasmid pGU366 containing urease gene was introduced into the B. subtilis SH14 with PEG-induced protoplast transformation (PIP) method. The urease gene was very stably expressed in the transformant of B. subtilis SH14. Also, the optimal conditions for transformation were established and the highest transformation frequency was obtained by treatment of lysozyme for 90 min, and then addition of 1.5 ${mu}g$/ml DNA and 40% PEG4000. From the in vitro antifungal test against F. solani, antifungal activity of B. subtilis SH14(pGu366) containing urease gene was much higher than that of the host strain. Genetical development of B. subtilis SH14 by transfer of urease gene can be responsible for enhanced biocontrol efficacy with its antibiotic action.
Three strains of common carp, i. e. , Israeli carp, red-and-white, and golden strains, were stocked in the same pond, and their growth rates were compared with following results: From August 12 to November 21 in 1975, fingerlings of the three strains of common carp, Cyprinus carpo, each weighing about 0.5 g with total length of 2 to 3 cm, were stocked. The pond had an area of $316m^2$ with a mean water depth of 55cm, and the bottom was covered with a 20 to 30 cm thick layer of silt containing a considerable amount of decaying organic materials. Feed given was prepared with equal amounts of fish meal and polished barley, of which, in addition, $10\%$ green grass and $1\%$ table salt were mixed together when prepared into paste feed using a chopper after boiling the barley. Total protein content of the feed was $34.9\%$ in dry state with $5\%$ moisture content. Total feed given was 30.08 kg calculated in dry state to produce 20.588 kg of the common carp fingerlings, thus the feed coefficient being 1.51. By strains, the harvested Israeli carp ranged 98 to 311g each with a mean weight of $172.69g(100\%)$, red-and-white strain 15 to 318g with mean of $104.1g(60.3\%)$, and the golden strain 30 to 268g with mean of $128.7g(74.6\%)$. During the rearing season mean water temperature was $23.9^{\circ}C$ and the assumed main growth period with the water temperature above $15^{\circ}C$ was, upto the end of October, for 80 days with a mean water temperature of $23.9^{\circ}C$. Taking this main growth period as the basis for growth rate analysis, the mean daily increments, expressed as the attained body weight in times of the starting weight, become 1.075786 times (or the Israeli strain, 1.06901 times for the red-and-white strain, and 1.07185 times for the golden strain.
The purpose of this study was to evaluate the thermal expansion characteristics of injectable ther-moplasticized gutta-perchas and a Resilon. The materials investigated are Obtura gutta-percha, Diadent gutta-percha, E&Q Gutta-percha Bar and Epiphany (Resilon). The temperature at the heating chamber orifice of an Obtura II syringe and the extruded gutta-percha from the tip of both 23- and 20-gauge needle was determined using a Digital thermometer. A cylindrical ceramic mold was fabricated for thermal expansion test, which was 27 mm long, with an internal bore diameter of 3 mm and an outer diameter of 10 mm. The mold was filled with each experimental material and barrel ends were closed with two ceramic plunger. The samples in ceramic molds were heated in a dilatometer over the temperature range from $25^{\circ}C$ to $75^{\circ}C$. From the change of specimen length as a function of temperature, the coefficients of thermal expansion were deter-mined. There was no statistical difference between four materials in the thermal expansion in the range from $35^{\circ}C$ to $55^{\circ}C$ (p > 0.05). However, Obtura Gutta-percha showed smaller thermal expansion than Diadent and Metadent ones from $35^{\circ}C$ to $75^{\circ}C$ (p < 0.05). The thermal expansion of Epiphany was similar to those of the other gutta-percha groups.
This study was a quasi-experimental study of nonequivalent control group pretest- posttest design to investigate the effect of home rehabilitation exercise program on the physical and psychological functions of home stayed chronic hemiplegic stroke patients. The data were collected during the period of May 20th to August 15th, 200l. The subjects for this study were 40 hemiplegic stroke patients with the experimental group consisting of 19 patients and the control group being composed of 21 patients. The patients selected for this study were: (a)living in J city who had been diagnosed with stroke and at home after being discharged from the hospital, (b) suffering from stroke for 6 months to 5 years, (c) without recognition disorder with the MMSE-K(Mini-Mental State Examination-K)score above 25, (d) below 2 on the modified Ashworth scale, (e)free from heart and pulmonary disease, (f)able to walk beyond 15 minutes for themselves, (g) not taking regular exercises. The program for the experimental group provided 8 weeks' home rehabilitation exercise, two times of group education during the first week and individual education and supportive care after the second week through home visiting and telephoning more than once a week. The amount of time spent on rehabilitation exercise by the experimental group was 35 to 50 minutes a day, three times a week. In order to understand the effects of experiment the two groups were compared and verified by measuring the physical and psychological functions of both groups. The data were analysed by $\chi^{2}-test$, paired t-test and unpaired t-test and ANCOVA through SAS/PC program. The results of the study were as follows: 1. In terms of physical variables: grip strength. lower extremity muscle strength, walking time, ADL and serum lipid levels 1) There was no significant difference in the unaffected and affected grip strength between the two groups, even though the unaffected and affected grip strength was more improved in the experimental group than in the control group. 2) There was no significant difference in the unaffected lower extremity muscle strength between the two groups, even though the unaffected lower extremity muscle strength was more improved in the experimental group than in the control group. There was no significant difference either in the affected lower extremity muscle strength between the two groups, even though the affected lower extremity muscle strength was more improved in the experimental group than in the control group. 3) There was significant difference in walking time between the two groups. Walking time was significantly reduced in the experimental group whereas it increased in the control group. 4) There was significant difference in ADL score between the two groups. ADL score was significantly increased in the experimental group, but it significantly decreased in the control group. 5) There was significant difference in serum total cholesterol level between the two groups. After experiment the serum T-C level became lower in the experimental group whereas it became sigficantly higher in the control group. 2. In terms of psychological variables: depression and self-esteem 1) There was no significant difference in the depression between the two groups, even though the depression showed constant in the experimental group, but it showed a significant increase in the control group. 2) There was no significant difference in the self-esteem between the two groups, even though the self-esteem showed some increase in the experimental group, but it significant decrease in the control group. As shown above, the results of 8 weeks' home rehabilitation exercise program for chronic hemiplegic stroke patients produced positive effects on walking time, ADL score and serum T-C level, shortening walking time, improving activities of daily living(ADL) and lowering serum total cholesterol level.
Journal of the Korean Society of Food Science and Nutrition
/
v.32
no.1
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pp.35-41
/
2003
The study was conducted to develope a rapid method for the detection of Yersinia enterocolitica in spring water and vegetables via multiplex polymerase chain reaction (PCR) technique using ail, yst, uirF and subgenus-specific Y16S primers. Specificity and sensitivity of multiplex PCR and application of best primers for the detection of Y. enterocolitica from spring water and vegetables were investigeted. Y. enterocolitica ATCC 27729 strains gave 356 bP and 200 bp (Y16S) and 134 bp (yst) bands. but Y. enterocolitica ATCC 9610 and ATCC 23715 strains gave 200 bp and 134 bp bands.In the meanwhile, non-pathogenic Yersinia species, such as Y. frederikseni, Y. inter-media, Y. kristenseni and Y. pseudotuberculosis gave only single 200 bp band, and other bacteria including Escherichia coli O157:H7 ATCC 25392, Shigella dysenteri. Staphylococcu aureus ATCC 25923 and Listeria mo-nocytogenes ATCC 19111 did not show any bands. Among primers, yst and Y16S primer showed the best sensitivity. Seven CFU/mL Y. enterocolitica cells could be detected with yst and Y16S primers and the sensitivity was significantly improved by the further 2nd PCR after 38 cycles of first PCR amplication. Spring water, cabbage and mushroom were inoculated with Y. enterocolitica to determine the sensitivity of multiplex-PCR for the rapid detection of Y. enterocolitica. Multiplex-PCR assay could detect 7 or 70 cells in spring water and vegetables using whole cell lysate with repeating PCR amplication.
In the analysis of variance between population and between individual trees (families), the fluctuation of values of variances due to sample size, (number of family) was analysed by two different designs, i.e. 2-level nested design with equal sample size and randomized complete block design. The variables were seedling heights and root calipers of 1-0 and 1-1 seedlings of Pinus densiflora S. et Z. The details of three natural stands and their progeny characters were presented in previous reports. 1. In nested design analysis. increase of sample size resulted the decrease of F-values among families in general, however, the F-values among populations showen the increasing tendency. The smaller the sample size, the larger the F-values fluctuation was resulted in general. At the point of beyond sample size 10, however, the fluctuation become to be stabilized. The F-value fluctuation seemed to be more in the case of analysis with random sampling method than with sequentially accumulated sampling method. And also such a tendency was more obvious in smaller sample size than in large one. 2. In R.C.B.D. analysis, the sample size to affirm the family variation was smaller than that for population variations.
Journal of the Korean Academy of Child and Adolescent Psychiatry
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v.5
no.1
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pp.70-82
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1994
Present study was to evaluate the validity and the clinical utility of the Korean version of Luria-Nebraska Neuropsychological Battery for Children(LNNB-C) in various groups including normal, brain damaged attention deficit hyperactivity disordered(ADHD), and psychiatrically disordered. The Korean version of LNNB-C and BGT were administered to clinical groups consisted of 51 patients(19 brain damaged, 16 ADHD. and 16 psychiatric controls), and to normal group composed of 147 children between the age of 8 and It Also KEDI-WISC was administered D clinical groups as a part of comprehensive psychological assessment There were significant differences between the brain damaged and the normals on all scales of LNNB-C, and between the normals and the ADHD on 11 clinical scales and 3 summary scales, which indicate the clinical validity for the scales of the Korean version of LNNB-C. The significant differences between the ADHD and the brain damaged on 3 summary scales were found, suggesting that the summary scales might play an important role id discriminating between two groups. Multiple discriminant analysis showed that the Korean version of LNNB-C significantly discriminates 3 groups - normals, ADHD, and brain damaged. Percentages of correct classification were ranged from 62.5% in the ADHD to 98.6Ta in the normals. For further evaluating the discriminant validity of the LNNB-C, the discriminant power of each items were calculated, and 131 of the 147 items discriminated significantly between the brain damaged and the normals. The scales of LNNB-C significantly correlated with the error scores of BGT and the most of scales of KEDI-WISC. These results put together : strongly support the concurrent and the discriminant validity of the Korean version of LNNB-C in diagnosing brain damage. The limitations of present study and several issues for the luther study were discussed.
Objective : The purpose of this study was to investigate the effect of Bee Venom and Melittin Solution on the lipopolysaccharide(LPS) and sodium nitroprusside(SNP)-induced expression of prostaglandin $E_2(PGE_2)$, cyclooxygenase-2(COX-2), nuclear factor kappa B($NF-{\kappa}B$) and nuclear factor kappa B($NF-{\kappa}B$) dependent luciferase activity in RAW 264.7 cells, a murine macrophage cell line. Methods : The expression of PGE2 was determined by determination of $PEG_2$, COX-2 was by western blotting with corresponding antibodies, $NF-{\kappa}B$ was by gel mobility shift assay method and $NF-{\kappa}B$ dependent luciferase activity was investigated by luciferase assay in RAW 264.7 cells. Results : 1. LPS and SNP-induced expression of $PEG_2$ was significant after 24hour. 2. The 0.5, 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS-induced expression of $PEG_2$ and, the $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly SNP-induced expression of $PEG_2$ compared with control, respectively. The 0.5 and $1{\mu}g/mL$ of bee venom could not significantly inhibit SNP-induced expression of $PEG_2$ compared with control. 3. The $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS and SNP-induced expression of COX-2 compared with control, respectively. The 0.5 and $1{\mu}g/mL$ of bee venom inclined to decrease LPS and SNP-induced expression of COX-2 compared with control. 4. The 0.5, 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS and SNP-induced expression of $NF-{\kappa}B$ compared with control, respectively. 5. The 0.5, 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS-induced expression of $NF-{\kappa}B$ dependent luciferase activity and the 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly SNP-induced expression of $NF-{\kappa}B$ dependent luciferase activity compared with control, respectively. The $NF-{\kappa}B$ inhibitor also inhibited significantly LPS and SNP-induced expression of $NF-{\kappa}B$ dependent luciferase activity compared with control. 6. The 0.5, 1 and $5{\mu}g/mL$ of bee venom and the 5 and $10{\mu}g/mL$ of melittin solution inhibited significantly LPS + IFN-${\gamma}$, TNF-${\alpha}$ and LPS + TNF-${\alpha}$-induced expression of $NF-{\kappa}B$ dependent luciferase activity compared with control, respectively. The $NF-{\kappa}B$ inhibitor also inhibited significantly LPS and SNP-induced expression of $NF-{\kappa}B$ dependent luciferase activity compared with control. Conclusions : These results suggest the inhibitory action of bee venom and melittin solution on the inflammatory mediators such as $PEG_2$, COX-2 and $NF-{\kappa}B$.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.31
no.3
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pp.199-218
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2005
Purpose of Study: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. Materials and Methods: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with ${\beta}$-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells$(1{\times}10^6)$ or BDNF-Ad infected Schwann cells$(1{\times}10^6)$ were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. Results: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were $1.54{\pm}4.0{\times}10^6$ and $9.66{\pm}9.6{\times}10^6$. 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell $0.69\;{\mu}g/{\mu}l$ of DNA was detected and in BDNF-Adenovirus transfected Schwann cell $0.795\;{\mu}g/{\mu}l$ of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. Conclusion: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.
Background : In order to elucidate one of the pathogenic mechanisms of ARDS associated with pulmonary surfactant and oxidant injury, acute lung injury was induced by N-nitroso N-methylurethane (NNNMU). In this model, the role of phospholipase $A_2$ ($PLA_2$), surfactant, gamma glutamyl transferase (GGT) and morphology were investigated to delineate one of the pathogenic mechanisms of ARDS by inhibition of $PLA_2$ with high dose of dexamethasone. Method: Acute lung injury was induced in Sprague-Dawley rats by NNNMU which is known to induce acute lung injury in experimental animals. To know the function of the alveolar type II cells, GGT activity in the lung and bronchoalveolar lavage was measured. Surfactant phospholipid was measured also. $PLA_2$ activity was measured to know the role of $PLA_2$ in ARDS. Morphological study was performed to know the effect of $PLA_2$ inhibition on the ultrastructure of the lung by high dose of dexamethasone. Results : Six days after NNNMU treatment (4 mg/kg), conspicuous pulmonary edema was induced and the secretion of pulmonary surfactant was decreased significantly. In the acutely injured rats' lung massive infiltration of leukocytes was observed. At the same time rats given NNNMU had increased $PLA_2$ and GGT activity tremendously. Morphological study revealed bizarre shaped alveolar type II cells and hypertrophied lamellar bodies in the cytoplasm of the alveolar type II cells. But after dexamethasone treatment (20 mg/kg, for six days) in NNNMU-treated rats, these changes were diminished i.e. there were decrease of pulmonary edema and increase of surfactant secretion from alveolar type D cells. Rats given dexamethasone and NNNMU had decreased $PLA_2$ and GGT activity in comparison to NNNMU induced ARDS rats. Conclusion : Inhibition of $PLA_2$ by high dose of dexamethasone decreased pathological findings caused by infiltration of leukocytes and respiratory burst. Based on these experimental results, it is suggested that an activation of $PLA_2$ is the one of the major factors to evoke the acute lung injury in NNNMU-induced ARDS rats.
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