• Title/Summary/Keyword: Sp1-21

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In vivo multiplex gene targeting with Streptococcus pyogens and Campylobacter jejuni Cas9 for pancreatic cancer modeling in wild-type animal

  • Chang, Yoo Jin;Bae, Jihyeon;Zhao, Yang;Lee, Geonseong;Han, Jeongpil;Lee, Yoon Hoo;Koo, Ok Jae;Seo, Sunmin;Choi, Yang-Kyu;Yeom, Su Cheong
    • Journal of Veterinary Science
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    • v.21 no.2
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    • pp.26.1-26.14
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    • 2020
  • Pancreatic ductal adenocarcinoma is a lethal cancer type that is associated with multiple gene mutations in somatic cells. Genetically engineered mouse is hardly applicable for developing a pancreatic cancer model, and the xenograft model poses a limitation in the reflection of early stage pancreatic cancer. Thus, in vivo somatic cell gene engineering with clustered regularly interspaced short palindromic repeats is drawing increasing attention for generating an animal model of pancreatic cancer. In this study, we selected Kras, Trp53, Ink4a, Smad4, and Brca2 as target genes, and applied Campylobacter jejuni Cas9 (CjCas9) and Streptococcus pyogens Cas9 (SpCas9) for developing pancreatic cancer using adeno associated virus (AAV) transduction. After confirming multifocal and diffuse transduction of AAV2, we generated SpCas9 overexpression mice, which exhibited high double-strand DNA breakage (DSB) in target genes and pancreatic intraepithelial neoplasia (PanIN) lesions with two AAV transductions; however, wild-type (WT) mice with three AAV transductions did not develop PanIN. Furthermore, small-sized Cjcas9 was applied to WT mice with two AAV system, which, in addition, developed high extensive DSB and PanIN lesions. Histological changes and expression of cancer markers such as Ki67, cytokeratin, Mucin5a, alpha smooth muscle actin in duct and islet cells were observed. In addition, the study revealed several findings such as 1) multiple DSB potential of AAV-CjCas9, 2) peri-ductal lymphocyte infiltration, 3) multi-focal cancer marker expression, and 4) requirement of > 12 months for initiation of PanIN in AAV mediated targeting. In this study, we present a useful tool for in vivo cancer modeling that would be applicable for other disease models as well.

THE EFFECTS OF DIETARY CONSISTENCY ON THE TRABECULAR BONE ARCHITECTURE IN GROWING MOUSE MANDIBULAR CONDYLE : A STUDY USING MICRO-CONFUTED TOMOGRAPHY (성장 중인 쥐에서 음식물의 경도가 하악 과두의 해면골에 미치는 영향 : 미세전산화 단층촬영을 이용한 연구)

  • Youn, Seok-Hee;Lee, Sang-Dae;Kim, Jung-Wook;Lee, Sang-Hoon;Hahn, Se-Hyun;Kim, Chong-Chul
    • Journal of the korean academy of Pediatric Dentistry
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    • v.31 no.2
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    • pp.228-235
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    • 2004
  • The development and proliferation of the mandibular condyle can be altered by changes in the biomechanical environment of the temporomandibular joint. The biomechanical loads were varied by feeding diets of different consistencies. The purpose of the present study was to determine whether changes of masticatory forces by feeding a soft diet can alter the trabecular bone morphology of the growing mouse mandibular condyle, by means of micro-computed tomography. Thirty-six female, 21 days old, C57BL/6 mice were randomly divided into two groups. Mice in the hard-diet control group were fed standard hard rodent pellets for 8 weeks. The soft-diet group mice were given soft ground diets for 8 weeks and their lower incisors were shortened by cutting with a wire cutter twice a week to reduce incision. After 8 weeks all animals were killed after they were weighed. Following sacrifice, the right mandibular condyle was removed. High spatial resolution tomography was done with a Skyscan Micro-CT 1072. Cross-sections were scanned and three-dimensional images were reconstructed from 2D sections. Morphometric and nonmetric parameters such as bone volume(BV), bone surface(BS), total volume(TV), bone volume fraction(BV/TV), surface to volume ratio(BS/BV), trabecular thickness(Tb. Th.), structure model index(SMI) and degree of anisotropy(DA) were directly determined by means of the software package at the micro-CT system. From directly determined indices the trabecular number(Tb. N.) and trabecular separation(Tb. Sp.) were calculated according to parallel plate model of Parfitt et al.. After micro-tomographic imaging, the samples were decalcified, dehydrated, embedded and sectioned for histological observation. The results were as follow: 1. The bone volume fraction, trabecular thickness(Tb. Th.) and trabecular number(Tb. N.) were significantly decreased in the soft-diet group compared with that of the control group (p<0.05). 2. The trabecular separation(Tb. Sp.) was significantly increased in the soft-diet group(p<0.05). 3. There was no significant differences in the surface to volume ratio(BS/BV), structure model index(SMI) and degree of anisotropy(DA) between the soft-diet group and hard-diet control group (p>0.05). 4. Histological sections showed that the thickness of the proliferative layer and total cartilage thickness were significantly reduced in the soft-diet group.

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Induction of Sexual Stage and Colony Morphology of Some Isolates of Sclerotium rolfsii Causing Spotted Leaf Rot in Plants

  • Pandey, M.K.;Sarma, B.K.;Singh, U.P.
    • Mycobiology
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    • v.33 no.1
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    • pp.7-11
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    • 2005
  • Twenty-two isolates of Sclerotium rolfsii causing spotted leaf rot from Varanasi, India were grown on 6% Cyperus rotundus rhizome meal agar (CRMA) medium for the induction of athelial stage (Athelia rolfsii). Only one isolate obtained from Sphaeranthus indicus formed basidial stage on CRMA medium while the other 21 isolates did not. Basidial stage was also produced in S. indicus isolate at different concentrations (5.5, 6.0 and 6.5% w/v) of CRMA medium. Size of basidia, sterigmata and basidiospores of this isolate was measured. Basidia clavate, hyaline and measured $10{\sim}12{\times}4{\sim}5\;{\mu}m$ in size, basidiospores hyaline, unicellular, subglobose to ellipsoid produced on sterigmata and measured $3{\sim}5{\times}2{\sim}4\;{\mu}m$ in size, sterigmata hyaline and measured $4{\sim}5{\times}1.5{\sim}2\;{\mu}m$ in size. The results of the present study revealed wide variation in spotted leaf rot isolates of S. rolfsii. A reddish zone around the colony of S. rolfsii isolate from Vernonia sp. was observed on CRMA medium. HPLC analysis of the zone revealed the presence of gallic and ferulic acid which were also thought to be responsible for reduced mycelial growth of the isolate on CRMA medium.

Kinetic Study on the Enzymatic Production of D-Alanine from D-Aspartic Acid

  • Lee, Jae-Heung;Sung, Moon-Hee;Jeon, Yeong-Joong
    • Journal of Microbiology
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    • v.40 no.1
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    • pp.33-37
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    • 2002
  • An enzymatic reaction for the production of D-alanine from D-aspartic acid and pyruvate as substrates by a thermostable D-amino acid aminotransferase (D-AAT) was investigated at various conditions In the temperature range of 40-70$\^{C}$ and pH range of 6.0-9.5. The D-AAT was produced with recombinant E. coli BL21, which hosted the chimeric plasmid pTLK2 harboring the D-AAT from the novel thermophilic Bacillus sp. LK-2. The enzyme reaction was shown to follow the Ping Pong Bi Bi mechanism. The K$\_$m/ values for D-aspartic acid and pyruvate were 4.38 mar and 0.72 mM, respectively. It was observed that competitive inhibition by D-alanine, the product of this reaction, was evident with the inhibition constant K$\_$i/ value of 0.1 mM. A unique feature of this reaction scheme is that the decorboxylation of oxaloacetic acid, one of the products, spontaneously produces pyruvate. Therefore, only a catalytic amount of pyruvate is necessary for the enzyme conversion reaction to proceed. A typical time-course kinetic study skewed that D-alanine up to 88 mM could be produced from 100 mM of D-aspartic acid with a molar yield of 1.0.

Zigbee based 1-ch ECG device with activity monitoring function (지그비를 기반으로 한 운동감시 기능을 가진 1채널 ECG장치 개발)

  • Hong, Joo-Hyun;Kim, Nam-Jin;Cha, Eun-Jong;Lee, Tae-Soo
    • Proceedings of the KIEE Conference
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    • 2005.10b
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    • pp.41-43
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    • 2005
  • PDA-based monitoring is used to acquire continuously the patient's vital signs, including electrocardiography, activity, heart rate and $SpO_2$. In this paper, A biomedical signal acquisition device was designed using 3-axial MEMS accelerometer and 1-ch ECG amplifier, to have the function of monitoring activity and electrocardiography. The proposed system is composed of transmitter and receiver. Through the Zigbee communication, subject's biosignals can be transmitted in real-time to receiver, and transmitted data confirmed using PDA. The packet size used in this device was set not to exceed a maximum payload size of 116 byte. One packet consists of two segments. The transmission speed was 21 packet per second, 420 ECG samples per second, and 42 acceleration samples per second. The proposed method can be used to develop Activities of Daily Living(ADL} monitoring devices for the elderly or movement impaired people and enables patients to be monitored without any constraints. Also, this method will reduce medical costs in the aged society.

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Effects of temperature on the biomass yield and the chemical composition of pseudomonas cells in continuous culture (Pseudomonas sp.의 연속배양에 있어서의 세포의 수율 및 화학적 조성에 미치는 영향)

  • 김창진;이영록
    • Korean Journal of Microbiology
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    • v.21 no.3
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    • pp.163-169
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    • 1983
  • Effects of temperature on the gorwth characteristics and the chemical composition of pseudomonas cells grown under glucose-or methanol-utilizing continuous culture were studied. In a glucose-utilizing continuous culture, optimum dilution rate, agitation, pH, and temperature, for the higher biomass yield were $0.45hr^-$, 7000rpm, pH 7.5, and $30^{\circ}C$, respectively. But in a methanol-utilizing continuous culture, they were $0.125hr^-$, 600rpm, pH 8, and $30^{\circ}C$, respectively. In methanol-utilizing continuous culture, the maximum production rate of the cells was 1.48g, dry wt./1/hr at a dilution rate of $0.45hr^-$, and the cell yield was 0.46g. dry wt./g. glucose. In the methanol-utilizaing continuous culture, the maximum production rate of the cells was 0.33 7g. dry wt./1/hr. at a dilution rate of $0.125hr^-$ and the cell yield was 0.44g dry cell/g. methanol. The contents of protein of the cells increase with the increase ingrowing temperature (from 15 to $30^{\circ}C$), more or less, while the contents of RNA nad carbohydrate of the cells decreased. However, DNA contents of cells growth under the various temperature ranges didn't change. As the temeprature of cultivation rises at a constant dilution rate, the efficiency of RNA in protein synthesis was increased, showing the decreases in the ratio of RNA to protein.

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Isolation of Yersinia enterocolitica from Springs water in Pusan Area (부산지역 약수터수로부터 Yersinia enterocolitica의 분리에 관한 연구)

  • Kim, Mi-Hee;Cha, In-Ho;Choi, Chul-Soon;Lee, Sang-Joon
    • Journal of Life Science
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    • v.7 no.3
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    • pp.219-227
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    • 1997
  • On the purpose pf epidemiological study related to yersiniosis, a total of 720 aprings water collected from 60 points in the Pusan area were examined for the presence of Y. enterocolitica and also the isolation rates, biotype, serotype, biochemical properties and antibiotic susceptibility. Fifty-eight(8.0%) strains of Yersinia species were isolated from 720springs water. Isolation rate for each species was 49 (6.8%)strains of Y. enterocolitica, 3 (0.4%) strains of Y. pseudotuberculosis and 6 (0.8%) strains of y. frederiksenii. Seasonal distribution of isolated Yersinia sp. were shown considerably from November to April, and Y. enterocolitica was especoally isolated in order of January (20.4%), December (16.3%), March(14.3%) and April(8.2%). Isolated T. enterocolitica was divided into 4 kinds of biotype such as 1, 2, 3, and 3B. Distribution of each biotype was shown in order of biotype 1 (51.0%), biotype 2 (30.6%), biotype 3B (16.3%) and biotype 3 (2.1%). The serotypes pf 49 Y. enterocoliticawere typed 7 kinds of werotypes (0 ; 3, 0 : 5, 0 : 9, 0 : 13, 0 : 18 and 0 : 21), and serotype 0 : 8(34.7%). 0 : 9(30.6%) and 0 : 3(10.2%) were encountered most frequently.

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Effects of dietary supplementation with fermented spent mushroom substrates of the winter mushroom (Flammulina velutipes) on growth performance, carcass traits, and economic characteristics of Hanwoo steers (발효 팽이버섯 수확후배지의 급여 수준이 한우 거세우의 비육과 도체성적 및 경제성 분석에 미치는 영향)

  • Moon, Yea-Hwang;Cho, Woong-Ki;Kim, Hyun-Jung;Kim, Ji-Eun;Kim, Bo-Ram;Kim, Hye-Soo;Cho, Soo-Jeong
    • Journal of Mushroom
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    • v.15 no.4
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    • pp.223-228
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    • 2017
  • This study was conducted to investigate the effects of fermented spent mushroom substrates (F-SMS) of Flammulina velutipes on growth performance, carcass traits, and economic characteristics of Hanwoo steers. A yeast strain (Saccharomyces sp. UJ14) and Bacillus strain (Bacillus sp. UJ03) isolated from fresh spent mushroom substrates of Flammulina velutipes were used as probiotics to prepare F-SMS. Twenty-four Hanwoo steers (14 months old) were allocated to three dietary treatments via a randomized block design and were slaughtered at 30 months of age. These treatment groups included Control (TMR), T1 (TMR containing 10% of F-SMS) group, and T2 (TMR containing 30% of F-SMS). Body weight gain was not influenced by the experimental diets. DM and TDN intakes in the finishing period were significantly (p < 0.05) greater in group T1 than in other groups. CP intake was significantly (p < 0.05) greater in group T2 than in other groups during the whole experimental period. Among carcass traits, rib-eye area and back fat thickness tended to increase with F-SMS supplementation. The appearance rate (%) of a meat yield more than grade A was the highest in group T1. The net profits increased by 1.2% and 13.3% in groups T1 and T2, respectively. In conclusion, if a proper feeding program (including feed safety) can be ensured, spent mushroom substrates of Flammulina velutipes can prove to be a highly profitable feed source for Hanwoo steers.

Incidence of Parasitic Infections and Diseases in Rearing and Fattening Cattle Raising in Gyeongbug District (경북지방(慶北地方)의 육성우(育成牛) 및 비육우(肥育牛)에 있어서 기생충(寄生蟲)의 감염(感染)과 질병실태조사(疾病實態調査))

  • Lee, Cha Soo;Lee, Jae Hyun;Byun, Myung Dae;Park, Cheong Kyu;Lee, Hi Suk;Moon, Moo Hong
    • Korean Journal of Veterinary Research
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    • v.20 no.2
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    • pp.179-197
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    • 1980
  • A survey was made from March 1978 through Feb. 1979 to know infection rate of parasites and incidence of diseases in 1968 cattle of 240 rearing and fattening cattle herds (1746 Korean native cattle, 186 Holstein, 34 Charolais and 2 Hereford) raising in 28 cities and counties of Gyeongbug district. The results obtained are summarized as follows: 1. The rate of nematoda infection was about 49.0% in the examined cattle. These nematodes were identified as Mecistocirrus sp., Oesophagostomum sp., Bunostomum sp., Trichostrongylus spp., Strongyloides sp. and other 3 species. Infection rate of nematoda in fattening and breeding cattle was higher than that in rearing calves, 44.3% of the positive cattle were infected with 2 to 5 species, and incidence of mixed infection was high in fattening cattle. 2. The rate of coccidal infection was 10.9%. Eimeria $z{\ddot{u}}rni$, Eimeria bovis and Eimeria bukidnonensis were mainly found in the examined cattle. The infection rates of cocidia in Holstein, breeding Korean cows, rearing Korean calves and fattening Korean cattle were 27.3%, 15.8%, 11.2% and 9.1%, respectively. 3. The rate of trematoda was 14.6% with Fasciola spp. (11.2%), Eurytrema spp. (2.0%), and Paramphistomum spp. infections (1.4%). Breeding Korean cows, fattening Korean cattle, Holstein and rearing Korean calves indicated 47.4%, 26.4%, 18.2% and 6.3% incidence of trematoda infections, respectively. of trematoda positive cattle, 76.6% were infected with Fasciola spp., 4. The rate of Theileria and Babesia infection was 62.7% in the examined cattle. Of these positive cattle 5.4% were infected with both parasites, 57.3% with Theileria, and 1.3% of rearing Korean calves and 22.6% of Holstein were found to have parasitized erythrocytes over 1.0%. 5. The incidence of diseases in 1968 cattle of 240 herds was 564 cattle (28.7%) of 154 herds (64.2%). of the diseases observed of skin occurred in 354 cattle (18.0%) of 61 herds (25.4%), diseases of respiratory system in 121 cattle (6.1%) of 34 herds (14.2%), diseases of digestive system in 38 cattle (1.9%) of 22 herds (9.2%), diseases of eye in 21 cattle (1.1%) of 13 herds (5.4%), diseases of urogenital system in 8 cattle (0.4%) of 8 herds (3.3%) and the other diseases in 24 cattle (1.2%) of 22 herds (9.2%). 6. Diseases of skin in 1968 cattle of 240 herds werds were observed in the highest incidence. Ringworm was observed in 13.3% of the examined herds, rearing Korean calves and Holstein showed high incidence. Scabies caused by Chorioptes bovis and alopecia localis along with infestation of lice (Damalinia bovis, Linognathus vituli) were observed in fattening Korean cattlein winter. 7. Diseases of respiratory system occurred frequently in rearing Korean cattle and Holstein. In the diseases of digestive system acute indigestion occurred in fattening Korean cittle, enteritis in rearing Korean calves and Holstein, and coccidiosis in rearing Korean calves maninly. 8. Infectious keratoconjunctivitis occurred in a fattening Korean cattle herd, sterility (2.0%) in breeding Korean cows, and theileriosis in Holstein calves were also occurred. In addition, poisoning and heat stroke were observed in several cattle.

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Detoxification of PSP and relationship between PSP toxicity and Protogonyaulax sp. (마비성패류독의 제독방법 및 패류독성과 원인플랑크톤과의 관계에 관한 연구)

  • CHANG Dong-Suck;SHIN Il-Shik;KIM Ji-Hoe;PYUN Jae-hueung;CHOE Wi-Kung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.22 no.4
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    • pp.177-188
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    • 1989
  • The purpose of this study was to investigate the detoxifying effect on PSP-infested sea mussel, Mytilus edulis, by heating treatment and correlation between the PSP toxicity and the environmental conditions of shellfish culture area such as temperature, pH, salinity, density of Protogonyaulax sp. and concentration of inorganic nutrients such as $NH_4-N,\;NO_3-N,\;NO_2-N\;and\;PO_4-P$. This experiment was carried out at $Suj\u{o}ng$ in Masan, Yangdo in Jindong, $Hach\u{o}ng\;in\;K\u{o}jedo\;and\;Gamch\u{o}n$ bay in Pusan from February to June in $1987\~1989$. It was observed that the detection ratio and toxicity of PSP in sea mussel were different by the year even same collected area. The PSP was often detected when the temperature of sea water about $8.0\~14.0^{\circ}C$. Sometimes the PSP fox of sea mussel was closely related to density of Protogonyaulax sp. at $Gamch\u{o}n$ bay in Pusan from March to April in 1989, but no relationship was observed except above duration during the study period. The concentration of inorganic nutrients effects on the growth of Protogonyaulax sp., then effects of $NO_3-N$ was the strongest among them. When the PSP-infested sea mussel homogenate was heated at various temperature, the PSP toxicity was not changed significantly at below $70^{\circ}C$ for 60 min. but it was proper-tionaly decreased as the heating temperature was increased. For example, when the sea mussel homogenate was heated at $100^{\circ}C,\;121^{\circ}C$ for 10 min., the toxicity was decreased about $67\%\;and\;90\%$, respectively. On the other hand, when shellstock sea mussel contained PSP of $150{\mu}g/100g$ was boiled at $100^{\circ}C$ for 30 min. with tap water, the toxicity was not detected by mouse assay, but that of PSP of $5400{\mu}g/100g$ was reduced to $57{\mu}g/100g$ even after boiling for 120 min.

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