Streptomyces platensis YK-2, newly isolated from forest soil, produces transglutaminase (TGase), which catalyses an acyl transfer reaction between the primary grade amine and protein or $\gamma$-carboxyamide group of peptide bound glutamine residues. For a molecular genetic study of S. platensis, an effective transformation method was established by using a conjugal transfer of DNA from Escherichia coli to spores of actinomycetes. The highest transconjugation frequency of S. platensis was obtained on an MS medium containing 50 mM $MgCl_2$, using $5{\times}10^7\;E$. coli as a DNA donor and $1{\times}10^8$ spores without heat treatment as a host. We also identified that S. platensis contains a single attB site within an ORF encoding a pirin-homolog, and that its attB site sequence shows high homology to that of S. logisporoflavus. In addition, it was confirmed by phenotypic analyses of exconjugants that the introduction of heterologous DNA into the attB site of the S. platensis chromosome does not affect its morphological differentiation and TGase production.
Law, Fang Lin;Zulkifli, Idrus;Soleimani, Abdoreza Farjam;Liang, Juan Boo;Awad, Elmutaz Atta
Asian-Australasian Journal of Animal Sciences
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v.31
no.8
/
pp.1291-1300
/
2018
Objective: This experiment was conducted to investigate the effects of dietary crude protein (CP) level and exogenous protease supplementation on growth performance, serum metabolites, carcass traits, small intestinal morphology and endogenous protease activity in broiler chickens reared under a tropical climate. Methods: A total of 480 day-old male broiler chicks were randomly assigned to eight dietary treatments in a $4{\times}2$ factorial arrangement. The main effects were CP level (21.0%, 19.7%, 18.5%, or 17.2% from 1 to 21 days and 19.0%, 17.9%, 16.7%, or 15.6% from 22 to 35 days) and protease enzyme supplementation (0 ppm or 500 ppm). All experimental diets were fortified with synthetic feed-grade lysine, methionine, threonine and tryptophan to provide the minimum amino acid recommended levels for Cobb 500. Results: Reducing dietary CP linearly reduced (p<0.05) growth performance, serum albumin, total protein, and carcass traits and increased (p<0.05) serum triglycerides and abdominal fat. There was no consistent effect of reducing dietary CP on morphological parameters of the intestine and on the pancreatic and intestinal endogenous protease activity (p>0.05). Protease supplementation improved (p<0.05) feed conversion ratio, body weight gain, carcass yield and intestinal absorptive surface area. Conclusion: Protease supplementation, as measured by growth performance, intestinal morphology and carcass yield, may alleviate the detrimental effects of low protein diets in broiler chickens.
Objective: To evaluate stromal cells of the bone marrow microenvironment (BMM) in bone marrow trephine biopsy (BMTB) specimens, with a focus on fibronectin, tumor necrosis factor- alpha (TNF-${\alpha}$) and L-selectin in Non-Hodgkin's lymphoma (NHL) patients, before and after therapy. Materials and Methods: A total of 80 de novo NHL patients, 64 with B-cell lymphomas 80%, (follicular cell lymphoma (FCL) in 32, chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) in 12, and diffuse large cell lymphoma in 20) and 16 with T-cell lymphomas (20%) all diagnosed as T-Lymphoblastic lymphomas, were evaluated before and after therapy. For comparison, 25 age and sex matched BM donors, were included as a control group. BMTB material and BM aspirates were taken for morphological assessment of stromal cells, the plasma of these samples being examined for $TNF{\alpha}$ and L-selectin by ELISA, and fibronectin by radial immunodiffusion (RID). Results: BM stromal cells comprising reticular macrophages and fibroblasts were elevated in 53.3% of NHL cases at diagnosis, while BM fibronectin levels were decreased and BM $TNF{\alpha}$ and L-selectin were higher than in controls (p<0.05). In NHL cases, elevated values of BM $TNF{\alpha}$ and BM L-selectin were associated with signs of aggressive disease, including >1 extra nodal sites, detectable B symptoms, high grade, BM and CNS invasion, and a high International prognostic index (IPI) (p<0.05). Conclusion: BMM components, $TNF{\alpha}$, L-selectin and fibronectin, in NHL can be useful in evaluating disease activity, extent and response to treatment and as prognostic markers according to the IPI.
Although many guidelines for pancreatic cystic neoplasms focus on the management of intraductal papillary mucinous neoplasm of the pancreas (IPMN) at the highest oncological risk, there are many issues that surgeons need to consider at the time to plan the surgical procedures based on characteristics of IPMN subtypes, such as multiplicity of branch duct-IPMN (BD-IPMN) and intraductal spreading of main duct-IPMN (MD-IPMN). For multifocal BD-IPMN, partial pancreatectomy would be selected to remove BD-IPMN with predictors of malignancy, while the other lesions without risk factors can be left, although total pancreatectomy might be considered if the patients have a strong family history of pancreatic cancer. Partial pancreatectomy would be also adequate procedure for MD-IPMN if negative surgical margin for high-grade dysplasia or invasive carcinoma were achieved. It has become to be well-known that patients with BD-IPMN are at increased risk for developing not only IPMN-associated pancreatic ductal adenocarcinoma (PDAC) but also PDAC independent from the IPMN. Hence, the detection of a concomitant PDAC is also an important focus for strategies after resection of BD-IPMNs. Our recent analysis of patients after partial pancreatectomy for MD-IPMN with negative surgical margin identified an unexpected recurrence pattern, which we called "monoclonal skip" recurrence. MD-IPMN seems to be disseminated in the pancreatic ductal systems and MD-IPMN with identical genetic background was detected in the remnant pancreas even in a long time after index surgery. We proposed strategies of postoperative surveillance based on characteristics and natural history of each morphological subtype.
Yang, J.S.;Jung, H.J.;Xuan, Z.N.;Kim, J.H.;Kim, D.S.;Chae, B.J.;Han, In K.
Asian-Australasian Journal of Animal Sciences
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v.14
no.10
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pp.1450-1459
/
2001
These experiments were conducted to investigate the effects of different feeding and processing methods of diets on performance, morphological changes in the small intestine and nutrient digestibility of growing-finishing pigs. One-hundred fifty growing pigs ($Yorkshire{\times}Landrace{\times}Duroc$; initial body weight of $23.33{\pm}0.75kg$) and one-hundred twenty finishing pigs ($Yorkshire{\times}Landrace{\times}Duroc$; initial body weight of $59.22{\pm}0.56kg$) were used in Exp. 1 and Exp. 2, respectively. Pigs were grouped on the basis of body weight and gender, and randomly allotted into 6 different treatments with 5 replications in each treatment in a $2{\times}3$ factorial arrangement. Treatments were 1) dry feeding with a mash diet (DM), 2) dry feeding with a pelleted diet (DP), 3) dry feeding with an expanded crumble diet (DEC), 4) dry/wet feeding with a mash diet (WM), 5) dry/wet feeding with a pelleted diet (WP), and 6) dry/wet feeding with an expanded crumble diet (WEC). In Exp. 1 (growing phase), there was no significant difference in average daily gain (ADG) and average daily feed intake (ADFI) among treatments during the entire experimental period, but feed conversion ratio (FCR) was significantly (p<0.05) improved in pigs fed pelleted diets regardless of feeding method. FCR was best in pigs fed a DP diet and worst in pigs fed a WM diet. Pigs fed a pelleted diet showed a 6.2% or 4.0% improvement in FCR compared with those fed a mash diet or an expanded crumble diet. Water disappearance was not significantly affected by dry/wet feeding or feed processing. Significant differences in villus height were not found among treatments, but villus height tended to be improved by dry/wet feeding. Dry/wet feeding or feed processing did not affect crypt depth. Digestibilities of calcium and phosphorus were significantly (p<0.05) improved in pigs fed an expanded crumble diet compared with pigs fed mash diets. Especially, pigs fed a WEC diet digested 8.1% more P than those fed a DM diet. Feed cost per kg weight gain (FCG) tended to be increased by dry/wet feeding rather than dry feeding. In Exp. 2 (finishing phase), ADG and ADFI were not significantly different among treatments, but a significant difference in FCR was found among feed processing forms. The best FCR was obtained in pigs fed a pelleted diet. Pigs fed a DP diet showed a 11.3% improvement compared with those fed a DEC diet. Water disappearance was significantly (p=0.0408) decreased by feeding the mash diet. However, water disappearance was not affected by dry/wet feeding during the finishing period. The villus height and crypt depth were not significantly different among treatments. However, crypt depth tended to be decreased by dry/wet feeding at the mid part of the small intestine. Fat digestibility was improved by dry feeding rather than dry/wet feeding, and was improved by 4.8% by feeding pellet diets compared with expanded crumble diets. Except for carcass grade, carcass characteristics were not significantly (p<0.05) different among treatments. Carcass grade was the best in pigs fed a WP diet. Feed cost per kg weight gain (FCG) was significantly decreased in pigs fed a pelleted diet compared with those fed an expanded crumble diet, and tended to be decreased by dry/wet feeding. In conclusion, these studies suggest that feeding the pelleted diet to growing-finishing pigs can be beneficial in terms of FCR and production cost. Dry/wet feeding can be helpful for the maintenance of villus height, but may not be reflected in improved growth performance or reduction of production costs.
Ha, Ho-Kyung;Kim, Jin Wook;Han, Kyoung-Sik;Yun, Sung Seob;Lee, Mee-Ryung;Lee, Won-Jae
Journal of Dairy Science and Biotechnology
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v.35
no.3
/
pp.208-214
/
2017
The aims of this study were to manufacture a hydrolyzed goat milk protein (HGMP)/chitosan ologisaccharide (CSO) nano-delivery system (NDS) and to investigate the effects of production variables, such as sodium tripolyphosphate (TPP), HGMP, and CSO concentration levels, on the formation and physicochemical properties of the NDS. An HGMP/CSO NDS was produced using the ionic gelation method at pH 5.5. Transmission electron microscopy and a particle size analyzer were used to determine the morphological and physicochemical properties of NDSs, respectively. The size of the HGMP/CSO NDS decreased from 225 to 138 nm as HGMP and CSO concentration levels decreased. The NDS had a positive surface charge, with a zeta-potential value of +23 mV. The encapsulation efficiency (EE) of docosahexaenoic acid was enhanced as the HGMP concentration level increased. Additionally, increasing the concentration level of CSO resulted in an increase in the EE of resveratrol. The HGMP/CSO NDS exhibited good physical stability during freeze-drying. Thus, our findings showed that the HGMP/CSO NDS was successfully manufactured and that HGMP and CSO concentration levels were key factors affecting the physicochemical properties of the NDS.
Early development of porcine oocytes fertilized in vitro was examined in different culture conditions. Porcine ovaries were collected from local slaughter-house. Cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cystein, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for 42~44 hrs. The frozen-thawed spermatozoa were washed by centrifigation 2 times at 1, 500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, 1$\mu\textrm{g}$/ml streptomycin and 1ng/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to 2.5$\times$10$^{6}$ cells/ml motile sperm during fertilization in vitro. At 8hrs h after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine and 4 mg/ml BSA and cultured for 7 days. In first experiment, the mean numbers of oocytes collected from 20 ovaries were 674.4 oocytes, and 4.1(27.6), 12.5(84.0), 25.4(171.6) and 57.9%(390.8) for A, B, C and D grade in morphological classification. In the second experiment, when culture medium was supplemented with various concentrations of EGF, the proportions of oocytes cleaved were 56.9, 55.7, 61.9 and 54.7% for 0, 5, 10 and 20ng/ml EGF. The higher proportions(15.1%) of oocytes developed to morular stage were obtained at concentration of 10ng/ml than 0 and 5ng/ml EGF (P<0.05). However, the proportions of embryos developed to blastocyst stage were not significantly different among concentrations of EGF. In another experiment, when the medium supplemented with catalase was used, the proportions of oocytes cleaved were higher in the concentration of 0 unit (56.5%, 61/108) than 100 and 1, 000 unit/ml of catalase (P<0.05). Although the developmental capacity of embryos was improved by medium with 0 unit/ml compared with 100, 500 and 1, 000 units/ml of catalase in oocytes developed to morula and blastocyst stages, were not significantly different among concentrations of catalase.
Eurasian eagle owl (Bubo bubo kiautschensis) is a natural monument (No. 324) and is also classified as an endangered wildlife grade II the Ministry of Environment of Korea. Eurasian eagle owl is threatened by the human being. But the efficient rescue and treatment for Eurasian eagle owl are still problematic, due to lack of basic data. In this study, to obtain basic resources for establishing more efficient rescue system, we held a physical examination and measured size physical part. In addition, we analyzed blood sample and feces samples and autopsy. We conduct the physical examination with measuring tapes and scales for 16 live features and 11 dead features. After inhalation anesthesia take the blood samples from wing vein. Blood samples are analyzed in hematologic and chemistric methods. We collected the feces through individual floor of cages from 12 live features and conducted the feces analysis. Autopsy was conducted for 11 dead samples. In physical examination, the mean body weight and total length of live features was 2.26${\pm}$0.35kg, 61${\pm}$2.38cm, the mean body weight and total length of dead features was 1.57${\pm}$0.51kg, 59.31${\pm}$3.51cm. Mean PCV was 46.57${\pm}$4.97%, while mean TPP was 3.49${\pm}$0.57g/dL. In feces sample analysis, 9 of 12 owls(75%) were infested by gastrointestinal helminths. The infection rates were: Trematoda (41.7%), Capilaria sp.(66.7%), acanthocephalans (25%). In autopsy, we confirmed 11 features were infested by duplicated infection of more than 2 helminths. Result of this study will be used for establishing reference range for physical features, blood analysis in Eurasian eagle owl. Therefore, to obtain basic resources for establishing more efficient rescue system, we have to make long-term conservation plans for Eurasian eagle owl.
Journal of The Korean Association For Science Education
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v.38
no.4
/
pp.575-585
/
2018
In this study, the texts of science textbooks of the past 20 years were collected in order to systematically carry out researches on scientific languages and scientific terms that have not been noticed in science education. We have collected all the science textbooks from elementary school to high school in the 6th curriculum, the 7th curriculum, and the 2009 revised curriculum, and constructed a corpus comprising of 132 textbooks in total. Sequentially, a raw corpus, a morphological annotated corpus, and a semantic annotated corpus of science terms, were constructed. The final constructed science textbook corpus was named K-STeC (Korean Science Textbook Corpus). K-STeC is a semantic annotated corpus with semantic classification and classification of scientific terms, together with meta information of bibliographic information such as curriculum, subject, grade, and publisher, location information such as chapter, section, lesson, page, and sentence, and structure information such as main, inquiry activities, reference materials, and titles. Throughout the three-year study period, a new research method was created by integrating the know-how of the three fields of linguistic informatics, computer science and science education, and a large number of experts were put in to produce labor-intensive results. This paper introduces new research methodologies and outcomes by looking at the whole research process and methods, and discusses the possibility of future development of scientific language research and how to use the results.
PURPOSE. The objective of this study was to conduct an in vitro comparative evaluation of polished and laser-dimpled titanium (Ti) surfaces to determine whether either surface has an advantage in promoting the attachment of epithelial-like cells and fibroblast to Ti. MATERIALS AND METHODS. Forty-eight coin-shaped samples of commercially pure, grade 4 Ti plates were used in this study. These discs were cleaned to a surface roughness (Ra: roughness centerline average) of 180 nm by polishing and were divided into three groups: SM (n=16) had no dimples and served as the control, SM15 (n=16) had $5-{\mu}m$ dimples at $10-{\mu}m$ intervals, and SM30 (n=16) had $5-{\mu}m$ dimples at $25-{\mu}m$ intervals in a $2{\times}4mm^2$ area at the center of the disc. Human gingival squamous cell carcinoma cells (YD-38) and human lung fibroblasts (MRC-5) were cultured and used in cell proliferation assays, adhesion assays, immunofluorescent staining of adhesion proteins, and morphological analysis by SEM. The data were analyzed statistically to determine the significance of differences. RESULTS. The adhesion strength of epithelial cells was higher on Ti surfaces with $5-{\mu}m$ laser dimples than on polished Ti surfaces, while the adhesion of fibroblasts was not significantly changed by laser treatment of implant surfaces. However, epithelial cells and fibroblasts around the laser dimples appeared larger and showed increased expression of adhesion proteins. CONCLUSION. These findings demonstrate that laser dimpling may contribute to improving the peri-implant soft tissue barrier. This study provided helpful information for developing the transmucosal surface of the abutment.
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