• Title/Summary/Keyword: Live tissue

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Comparison of the Viability of Cryopreserved Fat Tissue in Accordance with the Thawing Temperature

  • Hwang, So-Min;Lee, Jong-Seo;Kim, Hyung-Do;Jung, Yong-Hui;Kim, Hong-Il
    • Archives of Plastic Surgery
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    • v.42 no.2
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    • pp.143-149
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    • 2015
  • Background Adipose tissue damage of cryopreserved fat after autologous fat transfer is inevitable in several processes of re-transplantation. This study aims to compare and analyze the survivability of adipocytes after thawing fat cryopreserved at $-20^{\circ}C$ by using thawing methods used in clinics. Methods The survival rates of adipocytes in the following thawing groups were measured: natural thawing at $25^{\circ}C$ for 15 minutes; natural thawing at $25^{\circ}C$ for 5 minutes, followed by rapid thawing at $37^{\circ}C$ in a water bath for 5 minutes; and rapid thawing at $37^{\circ}C$ for 10 minutes in a water bath. The survival rates of adipocytes were assessed by measuring the volume of the fat layer in the top layers separated after centrifugation, counting the number of live adipocytes after staining with trypan blue, and measuring the activity of mitochondria in the adipocytes. Results In the group with rapid thawing for 10 minutes in a water bath, it was observed that the cell count of live adipocytes and the activity of the adipocyte mitochondria were significantly higher than in the other two groups (P<0.05). The volume of the fat layer separated by centrifugation was also measured to be higher, which was, however, not statistically significant. Conclusions It was shown that the survival rate of adipocytes was higher when the frozen fat tissue was thawed rapidly at $37^{\circ}C$. It can thus be concluded that if fats thawed with this method are re-transplanted, the survival rate of cryopreserved fats in transplantation will be improved, and thus, the effect of autologous fat transfer will increase.

Heat-Killed and Live Enterococcus faecalis Attenuates Enlarged Prostate in an Animal Model of Benign Prostatic Hyperplasia

  • Choi, Young-Jin;Fan, Meiqi;Tang, Yujiao;Iwasa, Masahiro;Han, Kwon-Il;Lee, Hongchan;Hwang, Ji-Young;Lee, Bokyung;Kim, Eun-Kyung
    • Journal of Microbiology and Biotechnology
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    • v.31 no.8
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    • pp.1134-1143
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    • 2021
  • In the present study, we investigated the inhibitory effect of heat-killed Enterococcus faecalis (E. faecalis) and live E. faecalis on benign prostatic hyperplasia (BPH). The BPH rat model was established by administering male rats with testosterone propionate (TP, 5 mg/kg, in corn oil) via subcutaneous injections daily for four weeks after castration. The rats were divided into five groups: Con, corn oil-injected (s.c.) + DW administration; BPH, TP (5 mg/kg, s.c.) + DW administration; BPH+K_EF, TP (5 mg/kg, s.c.) + heat-killed E. faecalis (7.5 × 1012 CFU/g, 2.21 mg/kg) administration; BPH+L_EF, TP (5 mg/kg, s.c.) + live E. faecalis (1 × 1011 CFU/g, 166 mg/kg) administration; BPH+Fi, TP (5 mg/kg, s.c.) + finasteride (1 mg/kg) administration. In both of BPH+K_EF and BPH+L_EF groups, the prostate weight decreased and histological changes due to TP treatment recovered to the level of the Con group. Both of these groups also showed regulation of androgen-signaling factors, growth factors, and apoptosis-related factors in prostate tissue. E. faecalis exhibited an inhibitory effect on benign prostatic hyperplasia, and even heat-killed E. faecalis showed similar efficacy on the live cells in the BPH rat model. As the first investigation into the effect of heat-killed and live E. faecalis on BPH, our study suggests that heat-killed E. faecalis might be a food additive candidate for use in various foods, regardless of heat processing.

Application of ELISA for the Detection of Sulfamethazine Residue in Live Cattle

  • Lee, H.J.;Lee, M.H.;Han, In K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.3
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    • pp.378-381
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    • 2001
  • Sulfamethazine has been widely used in swine for prevention or treatment of infections. Recently, the safety of the drug to consumers has been questioned because of carcinogenic effects. To prevent unwanted drug residues entering the human food chain, both government authorities and industries have established extensive control measures. The demands for reliable, simple, sensitive, rapid and low-cost methods for residue analysis of foods are increasing nowadays. In this study, we established a rapid prediction test for the detection of cattle with violative tissue residues of sulfamethazine. The recommended therapeutic dose of sulfamethazine (withdrawal time, 15 days) was administered to each of 10 cattle. Blood was sampled before drug administration and during the withdrawal period. The concentration of sulfamethazine in plasma, determined by a semi-quantitative ELISA, was compared to that of an internal standard (10 ppb). The absorbance ratio of internal standard to sample (B/Bs) was employed as an index to determine whether drug residues in cattle tissues were negative or positive. That is, a B/Bs ratio less than 1 was considered residue positive and if larger than 1 was considered negative. All 10 plasma samples from non-treated cattle showed negative to sulfamethazine. Sulfamethazine was detected in plasmas of treated cattle until Day 7 of withdrawal period. The present study showed that the semi-quantitative ELISA could be easily adapted in predicting residues of sulfamethazine in live cattle.

A Body Condition Scoring System for Bali Cattle

  • Soares, F.S.;Dryden, G. McL.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.11
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    • pp.1587-1594
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    • 2011
  • Live weight, body length, hip and shoulder heights, heart girth, and metatarsal length were measured on 100 one to two years old Bali (Bos javanicus) bulls. Multiple regression of these measurements on live weight gave a prediction equation involving heart girth and body length (prediction $R^2$ = 0.845). These measurements were also used to derive several frame scores (FS). Live weight (Lwt) divided by FS was used as an index of body condition. Lwt/(length+hip height) was normally distributed and highly correlated with other normally-distributed condition indexes. This index was used to define five body condition scores. These were used to develop a five-point body condition scoring system in which the amount of fleshing over the vertebral processes, ribs, hindquarters, tail head, hooks, at the top of the neck, and the shoulders, the development of wrinkles in the skin above the hock and the neck, and the size of the dewlap, were used to describe the different body condition scores. Animals of score 1 had prominent hooks, shoulders, vertebrae and ribs, and hollow hindquarters and flat tailhead. Score 5 animals had rounded hindquarters, well-filled upper hind legs, small mounds of soft tissue were apparent on the tailhead, their hooks, necks, shoulders, vertebrae and ribs were well covered, and the dewlap was prominent.

DETERMINATION OF LONGISSIMUS MUSCLE AREA IN PIG WITH ULTRASONIC LINEAR ELECTRONIC SCANNER

  • Irie, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.5 no.2
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    • pp.229-235
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    • 1992
  • The usefulness of a portable linear electronic scanner. B-mode ultrasonic machine, was evaluated for estimating the longissimus muscle area from ultrasonic measurement of the muscle depth in 22 live pigs. The electronic scanner was easy to operate for muscle measurements in live pigs, which did not have to be held but were caged. The cross-sectional images of longissimus muscle and covering muscles and fat appeared on the monitor with grey scale in real time. It was easy to identify the ultrasonograms of fat and muscular tissues because the images differed in the degree of the grey scale. The longissimus muscle had less echogenic image than the other muscles. The boundary lines between first, second or third layers of backfat and the longissimus muscle were distinct on the ultrasonogram. The ultrasonic measurement at the shoulder was not acceptable because of the unstable measurements and the complex tissue structure. The repeatabilities for the measurements of longissimus muscle depth at one-half body length and last rib were acceptable. The simple correlation coefficients between ultrasonic estimates of the muscle depth in live pigs and the actual areas in the carcass, were 0.50 and 0.55 at the last rib and the one-half body length, respectively. Therefore, those positions were similarly suitable to measure. The method of electronic scanner for determining longissimus muscle area from the muscle depth was suitable for practical use in the field because of the simple and inexpensive technique.

Optical Biopsy of Peripheral Nerve Using Confocal Laser Endomicroscopy: A New Tool for Nerve Surgeons?

  • Crowe, Christopher S;Liao, Joseph C;Curtin, Catherine M
    • Archives of Plastic Surgery
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    • v.42 no.5
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    • pp.626-629
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    • 2015
  • Peripheral nerve injuries remain a challenge for reconstructive surgeons with many patients obtaining suboptimal results. Understanding the level of injury is imperative for successful repair. Current methods for distinguishing healthy from damaged nerve are time consuming and possess limited efficacy. Confocal laser endomicroscopy (CLE) is an emerging optical biopsy technology that enables dynamic, high resolution, sub-surface imaging of live tissue. Porcine sciatic nerve was either left undamaged or briefly clamped to simulate injury. Diluted fluorescein was applied topically to the nerve. CLE imaging was performed by direct contact of the probe with nerve tissue. Images representative of both damaged and undamaged nerve fibers were collected and compared to routine H&E histology. Optical biopsy of undamaged nerve revealed bands of longitudinal nerve fibers, distinct from surrounding adipose and connective tissue. When damaged, these bands appear truncated and terminate in blebs of opacity. H&E staining revealed similar features in damaged nerve fibers. These results prompt development of a protocol for imaging peripheral nerves intraoperatively. To this end, improving surgeons' ability to understand the level of injury through real-time imaging will allow for faster and more informed operative decisions than the current standard permits.

Mechanisms of tissue factor induction by Porphyromonas gingivalis in human endothelial cells

  • Kim, So-Hee;Jung, Ji-Yeon;Kim, Won-Jae;Kim, Ok-Joon;Kim, Young;Kang, In-Chol
    • International Journal of Oral Biology
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    • v.46 no.3
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    • pp.99-104
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    • 2021
  • Associations between periodontal infection and cardiovascular disease have been documented. Porphyromonas gingivalis is a well-established periodontal pathogen, and tissue factor (TF) is a key initiator of the coagulation cascade. In this context, P. gingivalis has been reported to enhance TF expression in human endothelial cells. The present study investigated the underlying mechanisms of TF induction by P. gingivalis in human umbilical vein endothelial cells. P. gingivalis increased TF expression in a dose- and time-dependent manner. Not only live bacteria but also glutaraldehyde-fixed bacteria increased TF expression to the same extent. However, sonicates of P. gingivalis did not induce TF expression. Cytochalasin D and SMIFH2, which are inhibitors of actin polymerization and actin nucleation, respectively, inhibited the TF expression induced by P. gingivalis. Finally, TF production was decreased or increased in the presence of various signaling inhibitors, including mitogen-activated protein kinases. These results suggest that P. gingivalis induces endothelial TF expression by a bacterial internalization-dependent mechanism and through diverse signal transduction mechanisms.

Evaluation of stability and potency of live attenuated rinderpest vaccine of lapinized-avianized tissue culture strain origin for the establishment of expiration period (가토화·계태화 우역 생백신의 유효기간 설정을 위한 안정성 및 효능 평가)

  • Yeh, Jung-Yong;Kim, Hyun-Ju;Nah, Jin-Ju;Park, Jee-Yong;Lee, Ji-Hye;Seo, Hyun-Ji;Kweon, Chang-Hee;Cho, In-Soo;Moon, Jin-San
    • Korean Journal of Veterinary Research
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    • v.49 no.3
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    • pp.201-205
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    • 2009
  • In this study, we evaluated the stability and potency of live attenuated rinderpest vaccines of lapinized-avianized tissue culture strain origin, which had been produced annually from 2005 to 2008. When immune responses to the vaccines were evaluated using two Holstein calves weighing 100~150 kg, neutralizing antibody titer of 1 : 16 was induced at 21 days post vaccination. When calves were also inoculated with vaccines lots that had been stored for 39 months at ${4^{\circ}C}$, same level of antibody titer was observed. Using the virus titer test, we found that all batches of the vaccine that had been kept for 3, 10, 15, 22, 27, 34, 39, and 45 months showed no significant loss of titers, and fulfilled the requirement necessary ($\geq$ 3 $logTCID_50$) to be used as the national rinderpest vaccine reserve in Korea. In this study, we demonstrated that stability and potency of the rinderpest vaccines were maintained over three years when kept at ${4^{\circ}C}$ storage. This indicates that it maybe feasible to extend the expiration period of this vaccine from one year to three years.

Development of a cell-laden thermosensitive chitosan bioink for 3D bioprinting

  • Ku, Jongbeom;Seonwoo, Hoon;Jang, Kyoung-Je;Park, Sangbae;Chung, Jong Hoon
    • Proceedings of the Korean Society for Agricultural Machinery Conference
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    • 2017.04a
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    • pp.107-107
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    • 2017
  • 3D bioprinting is a technology to produce complex tissue constructs through printing living cells with hydrogel in a layer-by-layer process. To produce more stable 3D cell-laden structures, various materials have been developed such as alginate, fibrin and gelatin. However, most of these hydrogels are chemically bound using crosslinkers which can cause some problems in cytotoxicity and cell viability. On the other hand, thermosensitive hydrogels are physically cross-linked by non-covalent interaction without crosslinker, facilitating stable cytotoxicity and cell viability. The examples of currently reported thermosensitive hydrogels are poly(ethylene glycol)/poly(propylene glycol)/poly(ethylene glycol) (PEG-PPG-PEG) and poly(ethylene glycol)/poly(lactic acid-co-glycolic acid) (PEG/PLGA). Chitosan, which have been widely used in tissue engineering due to its biocompatibility and osteoconductivity, can be used as thermosensitive hydrogels. However, despite the many advantages, chitosan hydrogel has not yet been used as a bioink. The purpose of this study was to develop a bioink by chitosan hydrogel for 3D bioprinting and to evaluate the suitability and potential ability of the developed chitosan hydrogel as a bioink. To prepare the chitosan hydrogel solution, ${\beta}-glycerolphosphate$ solution was added to the chitosan solution at the final pH ranged from 6.9 to 7.1. Gelation time decreased exponentially with increasing temperature. Scanning electron microscopy (SEM) image showed that chitosan hydrogel had irregular porous structure. From the water soluble tetrazolium salt (WST) and live and dead assay data, it was proven that there was no significant cytotoxicity and that cells were well dispersed. The chitosan hydrogel was well printed under temperature-controlled condition, and cells were well laden inside gel. The cytotoxicity of laden cells was evaluated by live and dead assay. In conclusion, chitosan bioink can be a candidate for 3D bioprinting.

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Cholestrol Content and Formation of Oxidized Cholesterols in Processed Squids

  • Hong, Jeung-Hoon;Ryu, Hong-soo;Kim, Heung-Bae
    • Preventive Nutrition and Food Science
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    • v.1 no.2
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    • pp.196-202
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    • 1996
  • The effect of cooking(boiling, steaming and baking0and drying on the cholesterol content and formation of oxidized cholesterols in quid(Japanese flying squid, Todarodes pacificus) was studied. Cholesterol content of live squid meat varied with the portion sampled, and results from spectrophotometric assay ranged from 263.2mg/1..g(mantle) to 315.8mg/100g(tentacle). The cholesterol levels analyzed by gas chromatography(GC) for squid samples were lower by 7% of total cholesterol for live squid meat and 24% for processed meat than those results by spectrophotometric assay. Cooking resulted in the decrease of the initial total cholesterol content of raw meat from 10%(boiling for 5min.) to 25%(steaming for 5min.) The amounts of cholesterol remaining after baking were 68% for microwave oven samples and 64% for convection oven samples. Drying of raw tissue caused the greater reduction in cholesterol content than cooking but showed no significant difference in samples stored for 6 weeks at 4$^{\circ}C$ and 2$0^{\circ}C$. Raw squid meats contained essentially no oxidized cholesterols, while the 22-hydroxychoesterol was detected in frozen meats. The additional oxidized cholesterols as cholestane-triol was indentified with 22-hydroxycholesterols in cooked samples. Sun dried meat stored at 4一 and 2$0^{\circ}C$ for 6 weeks had the three kinds of oxidized cholesterols such as 22-hydroxycholesterol, cholesta-3,5-dien-7-one and cholestane-triol.

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