• Title/Summary/Keyword: in vivo passage

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Pathogenesis of infectious bronchitis virus with different routes of inoculation and the effect of in vivo serial passage in nephropathogenicity using cloacal infection

  • Lee, Chang-Won
    • Korean Journal of Veterinary Service
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    • v.25 no.1
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    • pp.87-96
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    • 2002
  • In this study, we wanted to determine if the respirotropic JMK strain of infectious bronchitis virus(IBV), which has a spike glycoprotein gene that is 99% similar to the nephropathogenic Gray strain of IBV, could adapt and cause lesions in the kidney following intracloacal passage in chickens. Two day old specific pathogen free(SPF) cchickens were infected with Gray and JMK strains by the intraocular and cloacal route. Several tissue samples were collected at various times. Viruses were recovered from more tissues and earlier in the infection from chickens infected cloacally than chickens infected intraocularly. Virus was isolated from the kidney of chickens infected with Gray by the intraocular route and JMK by the intracloacal route, but not from chicken given JMK the intraocular route. Histopathologically, interstitial nephritis was observed in Gray infected chickens. However, viral RNA or antigen were not detected in the kidney by in situ hybridization and immunohistochemistry. We further passaged the JMK strain ten times in two day old SPF chickens using cloacal inoculation. We examined the virus titer and histopathological change in the kidney at each passage level. The amount of virus recovered from the kidney was stable throughout this serial passage and the passaged virus did not caused renal damage. Further, virus could not be isolated from the kidney when chickens were infected with the passaged virus by the intraocular route. We conclude that the JMK strain has a strict upper respiratory tract tropism since cloacal passage did not produce nephrotropism or nephropathogenicity.

Decreased Contact Inhibition in Mouse Adipose Mesenchymal Stem Cells

  • Jeon, Yunmi;Lee, Myung Sook;Cheon, Yong-Pil
    • Development and Reproduction
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    • v.16 no.4
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    • pp.329-338
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    • 2012
  • The proliferation of embryonic cells or adult stem cells in tissue is critically regulated during development and repair. How limited the proliferation of cells, so far, is not much explored. Cell-cell contact proliferation inhibition is known as a crucial mechanism regulating cell proliferation in vitro and in vivo. In this study we examined the characters of mouse subcutaneous adipose derived stem cells (msADSC) whether they lost or get contact inhibition during in vitro culture. The characters of msADSC growth after confluence were analyzed using confocal microscope and the expression profiles of contact inhibition related genes were analyzed according to the morphological changes using real-time PCR method. msADSC showed overlapping growth between them but not after passage 14. The cell shapes were also changed after passage 14. The expression profiles of genes which are involved in contact inhibition were modified in the msADSC after passage 14. The differentiation ability of msADSCs to adipocyte, chondrocyte and osteocyte was not changed by such changes of gene expression profiles. Based on these results, it is revealed that smADSC were characterized by getting of strong cell-cell contact inhibition after passage 14 but the proliferation and developmental ability were not blocked by the change of cell-cell contact proliferation inhibition. These finding will help to understand the growth of adipose tissue, although further studies are needed to evaluate the physiological meaning of the cell-cell contact proliferation inhibition during in vitro culture of msADSC.

Studies on in vivo Nitrate Reduction in Rye (Secale cereale L.) Seedlings Treated with 2,4-Dinitrophenol II. Effect of 2,4-Dinitrophenol on in vivo Nitrate Reductase Activity in the Roots of Rye Seedlings (2,4-Dinitrophenol을 처리한 호밀(Secale cereale L.) 유식물의 질산염 환원에 관한 연구 II. 호밀 유식물 뿌리의 질산염 환원효소 활성에 대한 2,4-Dinitrophenol의 영향)

  • 조규찬
    • Journal of Plant Biology
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    • v.34 no.4
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    • pp.283-288
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    • 1991
  • This work was carried out to determined the effect of 2,4-dinitrophenol(DNP) on in vivo nitrate reductase activity in the root of 6 day old rye (Secale cereale L.) seedlings. The nitrate reductase activity in the roots of 6 day old rye seedlings pretreated with 0.5 mM DNP was higher than that of the control group in all the experimental conditions. The optimal concentration of KNO3 for maximum nitrate reductase activity was 10 mM in both control and treated group. The nitrate reductase activity in the treatment of 10 mM KNO3 gradually increased for 4 h in both groups, and then maintained constantly. The nitrate reductase activity occurred per hour was highest at 1 h in both groups, while it was declined by large degrees as time goes on. The daily pattern of nitrate reductase activity was gradually decreased in both groups with the passage of day. The optimal pH for this experiment and a previous paper (Kwon et al., 1991), it was determined that the nitrate reductase activity in both roots and shoots of rye seedlings was increased by the treatment of 0.5 mM DNP, and particulary in both groups, the nitrate reductase activity in the roots of rye seedlings was higher than that in shoots of them.

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BONE FORMATION BY HUMAN ALVEOLAR BONE CELLS (사람 치조골세포를 이용한 골형성)

  • Choi, Byung-Ho;Park, Jin-Hyoung;Huh, Jin-Young;Oh, Jin-Rok
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.28 no.1
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    • pp.42-45
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    • 2002
  • Cultures of primary human alveolar bone-derived cells were established from alveolar bone chips obtained from normal individuals undergoing tooth extraction. These cells were expanded in vitro until passage 3 and used for the in vivo assays. Cells were loaded into transplantation vehicles, and transplanted subcutaneously into immunodeficient mice to study the capacities of human alveolar bone-derived cells to form bone in vivo. Transplants were harvested 12 weeks after transplantation and evaluated histologically. Of 10 human alveolar bone-derived cell transplants, two formed a bone-like tissue that featured osteocytes and mineral. Eight of the ten formed no osseous tissue. These results show that cells from normal human alveolar bone are capable of forming bone-like tissue when transplanted into immunodeficient mice.

Maturational Changes in Binding Capacity of Fowl Sperm to the Epithelium of the Sperm Storage Tubules during Their Passage through the Male Reproductive Tract

  • Ahammad, Muslah U.;Okamoto, S.;Kawamoto, Yasuhiro;Nakada, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.9
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    • pp.1199-1203
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    • 2011
  • The objective of this study was to examine the binding potential of sperm to the epithelium of the sperm storage tubules (SST) in vitro and in vivo to assess the functional maturation of fowl sperm. Sperm from the testis, epididymis, as well as the proximal, middle and distal vas deferens were incubated in vitro with either the uterovaginal junction (UVJ)- or infundibular tissue containing SST at $39^{\circ}C$ for 30 min. Aliquots of sperm were also artificially inseminated into the uteri of hens, and the UVJ and infundibulum were collected 24 h post artificial insemination (AI). After incubation and AI, tissues were washed to remove loosely adhered sperm and subjected to fluorescence staining with 4', 6-diamidino-2-phenylindole, dihydrochloride (DAPI) for counting the number of bound sperm per 0.25 mm2 of surface area. Sperm from the testis, epididymis, and the three segments of the vas deferens exhibited their differential (p<0.05) binding capacity, which increased gradually from the testicular to distal vas deferens sperm under both in vitro and in vivo conditions. Existing similar trend, sperm, regardless of their source had a lesser affinity to bind to the epithelium of the infundibular SST than to the UVJ-SST. These experimental results suggested that fowl sperm may undergo gradual changes in the process of functional maturation, whereby they gain the ability to bind to the epithelium of the SST during their passage through the male reproductive tract (MRT).

Effect of flaking on the digestibility of corn in ruminants

  • Kang, Hamin;Lee, Mingyung;Jeon, Seoyoung;Lee, Sang Moon;Lee, Ju Hwan;Seo, Seongwon
    • Journal of Animal Science and Technology
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    • v.63 no.5
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    • pp.1018-1033
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    • 2021
  • In this study, we aimed to assess the effect of flaking on the nutrient digestibility of corn grain in ruminants. In this regard, in vitro rumen fermentation, in situ rumen degradability, and in vivo metabolic experiments were performed. The automated gas production technique was used for the in vitro fermentation experiments. Six types of corn flakes with various degrees of gelatinization (32%, 41%, 48%, 66%, 86%, and 89%) were ground and incubated in rumen fluid to measure rumen fermentation characteristics and digestion rate. The in situ degradability of ground corn, whole corn, and corn flakes with 62% and 66% gelatinization was measured by incubation in the rumen of two cannulated Holstein cows. In vivo metabolic experiments were performed using 12 crossbred goats (29.8 ± 4.37 kg) using a 3 × 3 Latin square design. The dietary treatments consisted of ground corn and flaked corn with 48% or 62% gelatinization. In vitro experiments showed that as the degree of gelatinization increased, the digestion rate increased linearly, while the discrete lag time decreased linearly (p < 0.05). The effective rumen dry matter degradability, determined by in situ fermentation, was 37%p lower in corn flakes than ground corn, assuming a passage rate of 6%/h (p < 0.01), and there was no difference between the two flakes. In the in vivo experiment, there was no difference in dry matter intake, average daily gain, feed efficiency, and nitrogen utilization among the treatment groups (p > 0.05); however, the crude fat digestibility was lower for corn flakes than for ground corn (p < 0.05). To summarize, the rate of fermentation of corn flakes increased as the degree of gelatinization increased. However, non-ground corn flakes had lower rumen digestibility and did not improve in vivo apparent nutrient digestibility, compared with ground corn. In contrast to the assumption that flaked corn provides more energy to ruminant animals than ground corn, we conclude that the digestibility and energy value of corn flakes are lower than those of ground corn if mastication does not sufficiently reduce the particle size of corn flakes.

Porcine Splenic Hydrolysate has Antioxidant Activity in vivo and in vitro

  • Han, Kyu-Ho;Shimada, Kenichiro;Hayakawa, Toru;Yoon, Taek Joon;Fukushima, Michihiro
    • Food Science of Animal Resources
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    • v.34 no.3
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    • pp.325-332
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    • 2014
  • The antioxidant capacity of porcine splenic hydrolysate (PSH) was studied in vitro and in vivo. Peptide hydrolysates were prepared, using the proteolytic enzyme $Alcalase^{(R)}$. The molecular weights of PSH were 37,666, 10,673, 6,029, and 2,918 g/mol. Rats were fed a 5% (w/v) PSH diet, instead of a casein diet, for 4 wk. The food intake, body weight gain, and liver weight of rats in the PSH group were similar to those in the control (CONT) group. There were no differences in the serum total cholesterol, triglyceride, total protein, or albumin levels between PSH and CONT groups. However, the level of in vivo hepatic lipid peroxidation in PSH group was significantly lower than that in CONT. In vivo hepatic catalase and glutathione peroxidase activities in the PSH group were significantly higher than those in the control group. The in vitro protein digestibility of PSH was lower than that of casein. The in vitro trolox equivalent antioxidant capacity of PSH was significantly higher than that of the peptide hydrolysate from casein. The in vitro radical scavenging activities of PSH were significantly higher than those of the peptide hydrolysate from casein. The present findings suggest that porcine splenic peptides improve the antioxidant status in rats by enhancing hepatic catalase and GSH-Px activities, and indicate a potential mechanism of radical scavenging activity during gastrointestinal passage.

Establishment and characterization of an immortalized human dermal papilla cell line

  • Shin, Seung-Hyun;Park, Sang-Yoon;Kim, Moon-Kyu;Kim, Jung-Chul;Sung, Young-Kwan
    • BMB Reports
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    • v.44 no.8
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    • pp.512-516
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    • 2011
  • Establishment of immortalized human dermal papilla cells (DPCs) retaining the characteristics of DPCs would be a great help for hair researchers. We recently established a simian virus 40T (SV40T)-transformed human DP cell line (SV40TDPC). However, the cell line senesced around passage 25 and ceased proliferation. In this study, we introduced the human telomerase reverse transcriptase (hTERT) gene into SV40T-DPC and established an immortalized human DP cell line. The cell line, SV40T-hTERT-DPC, did not induce tumors when inoculated into nude mice. SV40T-hTERT-DPC maintained morphology of early passage DPCs, expressed markers of DPCs, and retained responses to Wnt/${\beta}$-catenin and bone morphogenic protein (BMP) signaling pathways known to be required for hair-inducing activity of DPCs. The data strongly suggest that SV40T-hTERT-DPC retains many characteristics of human DPCs in vivo without malignant transformation.

Perceived Dark Rim Artifact in First-Pass Myocardial Perfusion Magnetic Resonance Imaging Due to Visual Illusion

  • Taehoon Shin;Krishna S. Nayak
    • Korean Journal of Radiology
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    • v.21 no.4
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    • pp.462-470
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    • 2020
  • Objective: To demonstrate that human visual illusion can contribute to sub-endocardial dark rim artifact in contrast-enhanced myocardial perfusion magnetic resonance images. Materials and Methods: Numerical phantoms were generated to simulate the first-passage of contrast agent in the heart, and rendered in conventional gray scale as well as in color scale with reduced luminance variation. Cardiac perfusion images were acquired from two healthy volunteers, and were displayed by the same gray and color scales used in the numerical study. Before and after k-space windowing, the left ventricle (LV)-myocardium boarders were analyzed visually and quantitatively through intensity profiles perpendicular the boarders. Results: k-space windowing yielded monotonically decreasing signal intensity near the LV-myocardium boarder in the phantom images, as confirmed by negative finite difference values near the board ranging -1.07 to -0.14. However, the dark band still appears, which is perceived by visual illusion. Dark rim is perceived in the in-vivo images after k-space windowing that removed the quantitative signal dip, suggesting that the perceived dark rim is a visual illusion. The perceived dark rim is stronger at peak LV enhancement than the peak myocardial enhancement, due to the larger intensity difference between LV and myocardium. In both numerical phantom and in-vivo images, the illusory dark band is not visible in the color map due to reduced luminance variation. Conclusion: Visual illusion is another potential cause of dark rim artifact in contrast-enhanced myocardial perfusion MRI as demonstrated by illusory rim perceived in the absence of quantitative intensity undershoot.

Characteristics of Digestion Dynamics of Rice and Oat Straw Relating to Microbial Digestion in the Rumen of Sheep Given High-Concentrate Diets

  • Goto, M.;Morio, T.;Kojima, E.;Nagano, Y.;Yamada, Y.;Horigane, A.;Yamada, H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.9
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    • pp.1219-1227
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    • 2000
  • Rumination behavior, in vivo digestibility of cell wall constituents, particle size reduction in the rumen, and retention time in the digestive tract of sheep were examined using rice and oat straw as roughage sources. The in sacco digestibility, rumen fermentation, and microbial population and internal adenosine 5-triphosphate (ATP) content were also determined under feeding conditions of high-roughage and high-concentrate diets. Chewing number and time in rumination behavior were higher with rice straw than with oat straw, while the in sacco and in vivo DMD of rice straw were consistently lower than those of oat straw. Rice straw also showed higher frequency of thinner and longer particles in the rumen contents and lower retention time in the whole digestive tract as compared to those of oat straw. Rice straw was more effective to maintain the ruminal pH than oat straw, being reflected in higher internal ATP content of large-type protozoa on the high- concentrate diet. Changes in the ruminal microflora by shifting from the low- to the high- concentrate diet were also different between rice and oat straw.