• Title/Summary/Keyword: Live tissue

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Adult stem cell lineage tracing and deep tissue imaging

  • Fink, Juergen;Andersson-Rolf, Amanda;Koo, Bon-Kyoung
    • BMB Reports
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    • v.48 no.12
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    • pp.655-667
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    • 2015
  • Lineage tracing is a widely used method for understanding cellular dynamics in multicellular organisms during processes such as development, adult tissue maintenance, injury repair and tumorigenesis. Advances in tracing or tracking methods, from light microscopy-based live cell tracking to fluorescent label-tracing with two-photon microscopy, together with emerging tissue clearing strategies and intravital imaging approaches have enabled scientists to decipher adult stem and progenitor cell properties in various tissues and in a wide variety of biological processes. Although technical advances have enabled time-controlled genetic labeling and simultaneous live imaging, a number of obstacles still need to be overcome. In this review, we aim to provide an in-depth description of the traditional use of lineage tracing as well as current strategies and upcoming new methods of labeling and imaging.

Using Linear Body Measurements of Live Sheep to Predict Carcass Characteristics for Two Iranian Fat-tailed Sheep Breeds

  • Kiyanzad, M.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.5
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    • pp.693-699
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    • 2004
  • Live animal selection programs that favor animals with a minimum amount of carcass fat are used for improving breeding flocks of sheep. To predict carcass characteristics of live sheep using body measurements in breeding flocks, 200 male and female lambs of two fat-tailed Iranian sheep breeds (Moghani and Makui) were used. Depth of soft tissue over the 12th rib of the live animals was measured with ultrasound (ULGR) and with hypodermic needle (NGR). The height at withers (HW), body length (BL), circumference of heart girth (CH) and width of hooks (WH), were measured. All animals were slaughtered; carcasses were cut into joints and dissected. Breed had a significant effect on all of the live easurements. The Moghani breed showed a higher value for HW, CH, ULGR and NGR, compared to that of Makui. Except for soft tissue depths; ULGR, NGR and GR, the male lambs showed higher values in live and carcass measurements than females. Percentages of carcass, total fat and intermuscular fat in females were higher than that of male lambs. In spite of the higher amount of subcutaneous and intermuscular fat in female (which is usually used for their physiological need, such as pregnancy and lactation), the male lambs had a heavier fat-tail than females. There was a wide range of variation of percentage of total carcass fat and total chemical fat content of carcass in the two breeds. Eventually this wide variation could be use by animal breeders for selection of animals with a lesser amount of carcass fat. Live weight of lambs showed a relatively low correlation with percentage of carcass lean, total fat and subcutaneous and intermuscular fat. Total lean meat was predicted with relatively high coefficients of determination in the two breeds ($R^2$=0.61 and 0.89, respectively). Live weight and carcass traits were predicted using simple measurements, but with $R^2$ ranging from 0.53 to 0.93.

Development of Semen Transport System for Cryopreservation and Fertility in Bull Sperm (소 정자의 동결 및 수정능력 향상을 위한 정액운반법의 개발)

  • Lee, Sang-Hee;Song, Eun-Ji;Woo, Jea-Seok;Lee, Seung-Hwan;Kang, Hee-Seol;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.37 no.3
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    • pp.97-102
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    • 2013
  • The objective of this study was to develop of semen transport system for cryopreservation and fertility in bull sperm. The ejaculated semen were diluted with Triladyl containing 20% egg-yolk for transportation. Diluted semen was transported by three methods that there were wrapping tissue (Tissue), sinking under $30^{\circ}C$ water (Water) and sinking between warm water and air (Air) methods. Semen was transported within 2 hours in $0.3^{\circ}C$. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk. And frozen-thawed sperm were estimated with SYBR14/PI double stain for viability, FITC-PNA/PI double stain for acrosome reaction analysis and Rhodamine123 double stain for mitochondrial intact assessment. In results, live sperm (SYBR+/PI-) in Air treatment group ($43.3{\pm}4.7%$) was significantly (p<0.05) higher than other treatment groups (Tissue: $16.3{\pm}2.7%$ and Water: $27.5{\pm}3.1%$), dying sperm (SYBR+/PI+) in Air treatment group ($55.6{\pm}4.7%$) was significantly lower than other treatment groups (Tissue: $77.6{\pm}3.2%$ and Water: $67.6{\pm}3.3%$) (p<0.05). Acrosome reaction in Air treatment group ($0.2{\pm}0.1%$) within live sperm (PI negative region) was significantly (p<0.05) lower than other treatment groups (Tissue: $0.7{\pm}0.2%$ and Water: $0.5{\pm}0.1%$), the acrosome reaction in Air treatment group ($28.6{\pm}2.8%$) within all sperm also was significantly lower than other treatment groups (Tissue: $44.2{\pm}1.8%$ and Water: $36.2{\pm}2.0%$) (p<0.05). And mitochondrial intact in Air treatment group within live ($97.1{\pm}0.4%$) and all ($61.9{\pm}3.3%$) sperm were significantly higher than other treatment groups (Tissue: $85.2{\pm}3.3%$, Water: $87.8{\pm}2.9%$ within live sperm and Tissue: $49.28{\pm}3.7%$, Water: $42.0{\pm}3.1%$ within all sperm) (p<0.05). Therefore, we suggest that transportation by sinking method between warm water and air was beneficial to improvement of fertility in frozen-thawed in bull semen.

Detection of Near-membrane Calcium Ions in Live Tissues with a Two-Photon Fluorescent Probe

  • Shin, Yu-Na;Lim, Chang-Su;Tian, Yu Shun;Rho, Won-Young;Cho, Bong-Rae
    • Bulletin of the Korean Chemical Society
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    • v.31 no.3
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    • pp.599-605
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    • 2010
  • A two-photon fluorescent probe (ACaCL) that can detect near-membrane $Ca^{2+}$ is reported. This probe can be excited by 780 nm fs pulses, shows high photostability and negligible toxicity, and can visualize near-membrane $Ca^{2+}$ in live cells and deep inside live tissues by two-photon microscopy.

Application of ELISA for the Detection of Oxytetracycline Residue in Live Animals

  • Lee, H.J.;Lee, M.H.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.12
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    • pp.1775-1778
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    • 2000
  • Oxytetracycline has been widely used in the cattle industry to control pneumonia, shipping fever, foot rot, bacterial enteritis, and uterine infections. Extensive use of antibiotics in veterinary clinics has resulted in residues in tissue and bacterial resistance to antibiotics. To prevent unwanted drug residues from entering the human food chain, extensive control measures have been established by both government authorities and industries. The demands for reliable, simple, sensitive, rapid and low-cost methods for residue analysis of foods are increasing. In this study, we established a rapid test for tissue residues of oxytetracycline in cattle. The recommended therapeutic dose of oxytetracycline (withdrawal time, 14 days) was administered to 10 cattle. Blood samples were collected from each cow before drug administration and during the withdrawal period. The concentration of oxytetracycline in plasma, determined by a semi-quantitative ELISA, was compared to that of the internal standard, 10 ppb. The absorbance ratio of internal standard to sample (B/Bs) was employed as an index to determine whether the residues in cattle tissues were negative or positive. That is, a B/Bs ratio less than 1 was considered as residue positive and that greater than 1 as negative. Based on this criterion, all plasma samples from cattle were negative to oxytetracycline at pre-treatment. Oxytetracycline could be detected in the plasma treated cattle until day 14 post-treatment. The present study showed that the semi-quantitative ELISA could be easily adapted in predicting tissue residues for oxytetracycline in live cattle.

The safety of live VHSV immersion vaccine at a temperature-controlled culture condition in juvenile olive flounder, Paralichthys olivaceus

  • Yo-Seb, Jang;Soo-Jin, Kim;Su-Young, Yoon;Rahul, Krishnan;Myung-Joo, Oh
    • Journal of fish pathology
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    • v.35 no.2
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    • pp.225-230
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    • 2022
  • Viral hemorrhagic septicemia (VHS) is one of the most serious viral diseases affecting farmed olive flounder (Paralichthys olivaceus) in Asian countries. VHS, caused by viral hemorrhagic septicemia virus (VHSV), occurs in over 80 different cultured and wild fish species worldwide. Our previous study demonstrated that VHSV infection can be restricted by adjusting the water temperature to over 17℃ from the host optima. We confirmed that the effective VHSV immersion vaccine treatment was a tissue culture infection dose (TCID) of 105.5 TCID50/mL at 17℃. However, the safety of live VHSV immersion vaccines remains unclear. The objectives of this study were to 1) demonstrate the safety of the live VHSV immersion vaccine under co-habitant conditions and 2) estimate the pathogenicity of VHSV in live VHSV-vaccinated flounder at 10℃. No mortality was observed in olive flounder treated with the live VHSV immersion vaccine, and the vaccinated flounder challenged with VHSV did not transfer VHSV to naïve fish at 10℃ through cohabitation. VHSV titration was below the detection limit (< 1.3 log TCID50/mL) in live VHSV immersion vaccine-treated flounder challenged with VHSV at 10℃. This study demonstrated that flounder treated with the live VHSV immersion vaccine were resistant to VHSV infection, and the live vaccine was also safe for naïve fish even at a water temperature known to be VHS infectious.

Effect of iron on the proteolytic activity of live Uronema marinum (Ciliata: Scoticociliatida) measured by fluorescence polarization

  • Lee, Eun-Hye;Kwon, Se-Ryeon;Choi, Seung-Hyuk;Kim, Ki-Hong
    • Journal of fish pathology
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    • v.19 no.1
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    • pp.83-86
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    • 2006
  • Effect of iron on the extracellular proteolytic activity of live Uronema marium was determined by fluorescence polarization (FP) method. Supplementation of 0.5 and 5.0 μM iron significantly increased caseinolytic activity of live U. marinum. In contrast, supplementation of 50 μM iron showed no significant differences in FP values compared to the control. The present result suggests that iron in cultured water or skin tissue of olive flounder may influence on the penetration and establishment of U. marinum, correlating with modulation of extracellular protease activity of the ciliates.

Fluorescent detection of bacteria associated with gingival sulcus epithelium (DNA 형광 염색을 이용한 치은열구상피부착 세균에 관한 연구)

  • Shin, Seung-Yun;Lee, Sang-Hyun;Yang, Seung-Min;Kye, Seung-Beom
    • Journal of Periodontal and Implant Science
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    • v.38 no.4
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    • pp.639-644
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    • 2008
  • Purpose: The aim of this study was to compare the number of live and dead bacteria attached to, or within, the stratified squamous epithelium lining the tissue side of the gingival sulcus. Materials and Methods: A total of 50 patients was examined and classified into healthy or diseased sites according to inflammatory status of the gingival tissue. The surface of stratified squamous epithelium was removed by gentle scraping of the gingival sulcus with curettes. The cells were processed in the laboratory by density-gradient centrifugation to separate the epithelial cells from the loose bacteria and debris. The LIVE/$DEAD^{(R)}$ $BacLight^{TM}$ Bacterial Viability Kit was applied and the specimens were observed by an epifluorescent microscope and the number of bacteria was counted. Results: Live and dead bacteria were stained to green and red, irrespectively. Generally, the number of total bacteria in the diseased sites was significantly higher than in the healthy sites. The mean number of detected bacteria in the diseased sites was $58.6{\pm}36.0$ (red bacteria $10.4{\pm}9.2$ / green bacteria $48.2{\pm}30.5$), while it was $1.5{\pm}1.7$ in the healthy sites (red bacteria $0.1{\pm}0.3$ / green bacteria $1.4{\pm}1.5$). The percentage of red bacteria was $17.5{\pm}11.2%$ in the diseased sites and $2.0{\pm}5.8%$ in the healthy sites. Conclusion: The total number of bacteria in the diseased sites was significantly higher than that of the healthy sites. The ratio and the number of red bacteria were also significantly higher in the diseased sites.

Application of ELISA for the Detection of Penicillin Antibiotic Residues in Live Animal

  • Lee, H.J.;Lee, M.H.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.11
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    • pp.1604-1608
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    • 2000
  • Penicillin antibiotics such as penicillin G, ampicillin and amoxicillin have been widely used in the pig industry to control salmonellosis, bacterial pneumonia, and urinary tract infections. Extensive use of antibiotics in veterinary clinics has resulted in tissue residues and bacterial resistance. To prevent unwanted drug residues entering the human food chain, extensive control measures have been established by both government authorities and industries. The demands for reliable, simple, sensitive, rapid and low-cost methods for residue analysis of foods are increasing. In this study, we established a rapid prediction test for the detection of pigs with unacceptable tissue residues of penicillins. The recommended therapeutic doses of three penicillins, penillin G (withdrawal time, 7 days), ampicillin (withdrawal time, 7 days) and amoxicillin (withdrawal time, 14 days), were administered to three groups of 20 pigs each. Blood was sampled before drug administration and during the withdrawal period. The concentration of penicillins in plasma, determined by a semi-quantitative ELISA, were compared to that of internal standard, 4 ppb, which corresponded to the Maximum Residue Limit in milk. The absorbance ratio of internal standard to sample (B/Bs) was employed as an index to determine whether drug residues in pig tissues were negative or positive. That is, a B/Bs ratio less than 1 was considered residue positive, and larger than 1 negative. All 60 plasma samples from pigs were negative to three penicillins at pretreatment. Penicillin G could be detected in the plasma of the treated pigs until day 4 post-treatment and ampicillin until day 2, whereas amoxicillin could be detected until day 10 of its withdrawal period. The present study showed that the semi-quantitative ELISA could be easily adapted to detect residues of penicillin antibiotics (penicillin G, ampicillin and amoxicillin) in live pigs.