• Microbiology and Biotechnology Letters
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• v.37 no.4
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• pp.377-382
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• 2009

Viral safety is an important prerequisite for clinical preparations of all biopharmaceuticals derived from plasma, cell lines, or tissues of human or animal origin. To ensure the safety, implementation of multiple viral clearance (inactivation and/or removal) steps has been highly recommended for manufacturing of biopharmaceuticals. Of the possible viral clearance strategies, Ultraviolet-C (UVC) irradiation has been known as an effective viral inactivating method. However it has been dismissed by biopharmaceutical industry as a result of the potential for protein damage and the difficulty in delivering uniform doses. Recently a continuous flow UVC reactor (UVivatec) was developed to provide highly efficient mixing and maximize virus exposure to the UV light. In order to investigate the effectiveness of UVivatec to inactivate viruses without causing significant protein damage, the feasibility of the UVC irradiation process was studied with a commercial therapeutic protein. Recovery yield in the optimized condition of $3,000\;J/m^2$ irradiation was more than 98%. The efficacy and robustness of the UVC reactor was evaluated with regard to the inactivation of human immunodeficiency virus (HIV), hepatitis A virus (HAV), bovine herpes virus (BHV), bovine viral diarrhea virus (BVDV), porcine parvovirus (PPV), bovine parvovirus (BPV), minute virus of mice (MVM), reovirus type 3 (REO), and bovine parainfluenza virus type 3 (BPIV). Non enveloped viruses (HAV, PPV, BPV, MVM, and REO) were completely inactivated to undetectable levels by $3,000\;J/m^2$ irradiation. Enveloped viruses such as HIV, BVDV, and BPIV were completely inactivated to undetectable levels. However BHV was incompletely inactivated with slight residual infectivity remaining even after $3,000\;J/m^2$ irradiation. The log reduction factors achieved by UVC irradiation were ${\geq}3.89$ for HIV, ${\geq}5.27$ for HAV, 5.29 for BHV, ${\geq}5.96$ for BVDV, ${\geq}4.37$ for PPV, ${\geq}3.55$ for BPV, ${\geq}3.51$ for MVM, ${\geq}4.20$ for REO, and ${\geq}4.15$ for BPIV. These results indicate that UVC irradiation using UVivatec was very effective and robust in inactivating all the viruses tested.

• Journal of Chest Surgery
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• v.29 no.1
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• pp.7-13
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• 1996

The conventional glutaraldehyde (GA) fixation method of tissue valves is considered to be responsible for accelerated valve degeneration. The release of toxic GA from the valve tissue is believed to limit endothelial cell (EC) ingrowth. Removal of toxic GA by reaction with L-glutamic acid and storage in a Paraben solution may offer good EC growth. To investigate the conditions for endothelialization of tissue valves, the growth properties of ECs on the conventionally and alternatively treated pericardial tissue were compared. Conventional preparation included zero-pressure fixation for 72 hours in phosphated-buffered saline (PBS) solution containing 0.5% GA at 4$^{\circ}C$ and storage into PBS containing 0.2% GA(group I). Alternatively treated pericardial tissues were divided into three postfixation treatment groups : (1) storage in PBS solution containing Paraben(group II), (2) treatment with PBS containing 8$^{\circ}C$ L-glutamic acid(PH 7.35) and storage in PBS solution containing Paraben (g oup III), (3) treatment with L-glutamic acid dissolved in distilled water (PH 3.5) (group IV). Pericardial tissue were transferred into the 24-well plate after storage for 4 weeks. ECs were harvested enzymatically from the bovine pulmonary artery and grown to confluence on culture flask surfaces. Detached ECs by trypsin were incubated into the each well of the 24-well plate including test pericardial tissues. Cells were detached by trypsin, 1, 2, 3, 5, 7 days after incubation and counted on the hemacytometer. Cell viability test was performed by frypan-blue exclusion method. Acute cell death in the group I were found even after prolonged washing. The group II showed prolonged cell survival compared with the group I. Both group III and group IV showed better cell growth than group II. There was no statistically significant difference between group III and group IV method in terms of EC growth. This results suggest that treatment by L-glutamic ac id and storage in a Paraben solution be a promising approach for improvement of durability of GA-treated tissue valves.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.10
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• pp.1344-1349
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• 2004

Titin-cap (TCAP), one of the abundant transcripts in skeletal muscles, was nvestigated in this study in cattle because of its role in regulating the proliferation and differentiation of myoblasts by interacting with the myostatin gene. From the 5, and 3, RACE experiments, full-length TCAP coding sequence was identified, comprising 166 amino acids. The amino acid comparison showed high sequence similarities with previously identified human (95.8%) and mouse (95.2%) TCAP genes. The TCAP expression, addressed by northern blot, is limited in muscle tissues as indicated by Valle et al. (1997). The radiation hybrid analysis localized the gene on BTA19, where the comparative human and porcine counterparts are on HSA17 and SSC12. A few muscle-related genetic disorders were mapped on HSA17 and some growth-related QTLs were identified on SSC12. The bovine TCAP gene found in this study opens up new possibilities for the investigation of muscle-related genetic diseases as well as meat yield traits in cattle.

• Journal of Veterinary Clinics
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• v.23 no.3
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• pp.279-285
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• 2006

Thirty dairy cows with pododermatitis were selected and treatment effect of ozone ointment for bovine pododermatitis was investigated. In addition, bactericidal effect of ozone ointment on etiological agent of bovine pododermatitis was evaluated. The pathohistological examination for the pododermatitis, according to application with ozone ointment was investigated. Thirty dairy cows were divided two groups: control group(vaseline group: 15 cows), treatment group(ozone ointment group: 15 cows). Various parameters were evaluated in terms of the lameness score, swelling score, lesion score, WBC, neutrophil, pathohistological finding, and antimicrobial action. As compared with vaseline group, ozone ointment group revealed significant decrease of lameness(p<0.05), swelling(p<0.01) and lesion score(p<0.05) were shown in hoof lesions on 14 days after application. In hematological findings, WBC count revealed slightly high values within normal range before treatment, however, this was 1Corresponding author improved on 14 days after application of ozone ointment. The number of neutrophils was slightly higher than that of normal, however, this was improved on 14 days after application of ozone ointment. In pathohistological findings, normal dermal tissue was found in tissues with pododermatitis on 14 days after application of ozone ointment. In antimicobial action, marked decrease rate of bacteria was observed in feet of all cases treated with ozone ointment. The decreasing rate of bacteria in anaerobic culture was higher than that in aerobic culture.

• BMB Reports
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• v.28 no.3
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• pp.254-260
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• 1995

Cytochrome oxidase was purified from bovine-heart mitochondria and its enzymatic properties were examined. The purified cytochrome oxidase was identified by its absorption spectrum and chromatogram through gel filtration. The specific activity, purification degree and yield of purified cytochrome oxidase were 18 nmol/mg/ml/min, 24.83 fold and 0.93%, respectively. The activity of the enzyme assayed by a ferrocytochrome $c-O_2$ system was optimized at $25^{\circ}C$ and pH 6.5. Examining the effect of nonionic detergents established that cytochrome oxidase was deactivated by Triton X-100. The oxidase was activated by Tween 80 and deactivated by Tween 20. The Michaelis constant and maximum velocity of the oxidase for ferrocytochrome c were 0.032~0.044 mM and 0.019~0.021 mM/min, respectively. After adaption to basal diet for a week, experimental diets containing 6 mg Cu/kg, or zero mg Cu/kg, or 12 mg Cu/kg were fed to a control group, a copper-free group and a copper-rich group of Sprague-Dawley rats, respectively, for 4 weeks. The specific activities assayed for the ferrocytochrome $c-O_2$ system of isolated cytochrome oxidase from the rat liver of control, copper-free, and copper-rich group were 1.00, 1.19, and 0.878 nmol/mg/ml/min, respectively. Their degrees of purification were 11.38, 10.82 and 8.78 fold, respectively. The specific activities for liver and heart mitochondrial cytochrome oxidase of copper-free/copper-rich groups assayed using the ferrocytochrome $c-O_2$ system were 81.4% and 96.4%/64.1% and 61.1%, respectively, compared with those of the control.

• Journal of Chest Surgery
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• v.45 no.6
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• pp.368-379
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• 2012

Background: Bioprostheses for cardiovascular surgery have limitations in their use following as calicification. ${\alpha}$-galactosidase epitope is known as a stimulant of immune response and then shows a progressing calcification. The objective of this study was to evaluate histologic characteristics and mechanical properties of decellularization and treated with ${\alpha}$-galactosidase. Materials and Methods: Bovine pericardial tissues were allocated into three groups: fixation only with glutaraldehyde, decellularization with sodium dodesyl sulfate and decellularization plus treatment with ${\alpha}$-galactosidase. We confirmed immunohistological characteristics and mechanical properties as fatigue test, permeability test, compliance test, tensile strength (strain) test and thermal stability test. Results: Decellularization and elimination of ${\alpha}$-gal were confirmed through immunohistologic findings. Decellularization had decreased mechanical properties compared to fixation only group in permeability (before fatigue test p=0.02, after fatigue test p=0.034), compliance (after fatigue test p=0.041), and tensile strength test (p=0.00). The group of decellularization plus treatment with ${\alpha}$-galactosidase had less desirable mechanical properties than the group of decellularization in concerns of permeability (before fatigue test p=0.043) and strain test (p=0.001). Conclusion: Favorable decellularization and elimination of ${\alpha}$-gal were obtained in this study through immunohistologic findings. However, those treatment including decellularization and elimination of ${\alpha}$-gal implied the decreased mechanical properties in specific ways. We need more study to complete appropriate bioprosthesis with decellularization and elimination of ${\alpha}$-gal including favorable mechanical properties too.

• Preventive Nutrition and Food Science
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• v.7 no.4
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• pp.432-436
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• 2002

A proteomic map of Hanwoo loin was obtained using 2-D SDS-PAGE and mass spectrometric analysis: 27 bovine proteins plus 2 proteins having similarities to other mammal proteins out of 52 proteins analyzed. The identified proteins consisted of 50 % basic house keeping proteins involved in metabolism, 30% muscle proteins, and other miscellaneous proteins. Many proteins on the 2-D gel with different molecular weights and isoelectric points were identified as same proteins due to posttranslational modification. As many of the identified house keeping proteins showed the high sequence similarities to other mammal equivalent proteins, searching the mammal databases could confirm the annotation. The preliminary identification of the proteome in bovine loin tissue could reveal the functions of proteins at over 50 % of chance with high fidelities. Using the established loin proteome map, proteomic difference between 1 yr and 2 yr Hanwoo loin tissues were compared on 2D gel. Regardless of the difficulty normalizing protein concentrations and sample-to-sample variations, three unidentified proteins and myoglobin were selected as up-regulated proteins during the fat deposition period. This study contributes to a move thorough and holistic understanding of beef meat, helping to build the basis for future identification of new markers for good quality meat.

• Journal of Periodontal and Implant Science
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• v.32 no.3
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• pp.475-488
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• 2002

The purpose of this study is to evaluate histologically the resorption and tissue response of various resorbable collagen membranes used for guided tissue regeneration and guided bone regeneration, using a subcutaneous model on the dorsal surface of the rat. In this study, 10 Sprague-Dawley male rats (mean BW 150gm) were used and the commercially available materials included acellular dermal matrix allograft, porcine collagen membrane, freeze-dried bovine dura mater. Animals were sacrificed at 2,6 and 8 weeks after implantation of various resorbable collagen membranes. Specimens were prepared with Hematoxylin-Eosin stain for light microscopic evaluation. The results of this study were as follows: 1. Resorption : Inner portion of porcine collagen membrane was resorbed a lot at 6 weeks, but its function was being kept for infiltration of another tissues were not observed. Freeze-dried bovine dura mater and acellular dermal allograft were rarely resorbed and kept their structure of outer portion for 8 weeks. 2. Inflammatory reactions : Inflammatory reaction was so mild and foreign body reaction didn't happen in all of resorbable collagen membranes, which showed their biocompatibility. 3. In all of resorbable collagen membranes, multinuclcated giant cells by foreign body reactions were not observed. Barrier membranes have to maintain their function for 4-6 weeks in guided tissue regeneration and at least 8 weeks in guided bone regeneration. According to present study, we can find all of the resorbable collagen membranes kept their function and structure for 8 weeks and were rarely resorbed. Foreign body reaction didn't happen and inflammatory reaction was so mild histologically. Therefore, all of collagen membranes used in this experiment were considered proper resorbable membranes for guided tissue regeneration and guided bone regeneration.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.33 no.1
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• pp.28-39
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• 2007

The purpose of this study is to evaluate the regenerative capacity of reconstruction in the atrophied posterior maxilla by comparing bone graft procedures and alveolar distraction osteogenesis (ADO) techniques. We performed the autogenous iliac bone graft (AGB group, 5 specimens in 3 patients), and the combination (Mixed group, 3 specimens in 3 patients) of the autogenous and deproteinized bovine bone ($Bio-Oss^{(R)}$, Geistlich Co., Switzerland) as the ratio of 2:1 in the sinus floor elevation procedures. ADO procedures using $TRACK^{(R)}$ (KLS Martin Co., Germany) were also performed to augment vertical alveolar height in atrophied posterior maxilla (ADO group, 5 specimens in 4 patients). Newly generated bone tissues were obtained with the 2.0mm diameter trephine bur (3i Co., USA) during implant fixture installation after 5-7 months. Routine histolomorphological observation, immunodot blot assay for quantitative evaluation, and immunohistochemical staining with antibodies to MMP-1, -9, -10, TIMP-1, -2, and BMP-2, -4 were all carried out. Lamellar bone formation was well shown in all specimens and new bone formations of ADO group increased than those of other procedures. In immunohistochemical staining, the strong expression of BMP-2 was shown in all specimens, and immunodot blot assay showed that bone formation is accompanied by the good induction of factors associated with angiogenesis and appeared more increased amount of osteogenic and angiogenic factors in ADO group. ADO is the most effective technique for new bone formation compared to sinus floor elevation with autogenous or mixed bone graft in the atrophied posterior maxilla. In the quantitative immunodot blot assay, the regenerated bone after ADO showed more increased products of VEGF, BMP-2, PCNA and MMP-1 than those after the other procedures, and these findings were able to be confirmed by immunohistochemical stainings.

• Journal of Periodontal and Implant Science
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• v.32 no.1
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• pp.161-172
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• 2002

The present study evaluated the effects of guided tissue regeneration using xenograft material(deproteinated bovine bone powder), with and without biodegradable membrane in beagle dogs. Contralateral fenestration defects (6 ${\times}$ 4mm) were created 4 mm apical to the buccal alveolar crest of maxillary premolar teeth in 5 beagle dogs. Deproteinated bovine bone powders were implanted into fenestration defect and one randomly covered biodegradable membrane (experimental group). Biodegradable membrane was used to provide GTR. Tissue blocks including defects with soft tissues which were harvested following four & eight weeks healing interval, prepared for histo-phathologic analysis. The results of this study were as follows. 1. In control group, at 4 weeks after surgery, new bony trabecular contacted with interstitial tissue and osteocytes like cell were arranged in new bony trabecule. Bony lamellation was not observed. 2. In control gruop, at 8 weeks after surgery, scar-like interstitial tissue was filled defect and bony trabecule form lamellation. New bony trabecular was contacted with interstitial tissue but defect was not filled yet. 3. In experimental group, at 4 weeks after surgery, new bony trabecular partially recovered around damaged bone. But new bony trabecular was observed as irregularity and lower density. 4. In experimental group, at 8 weeks after surgery, lamella bone trabecular developed around bone cavity and damaged tissue was replaced with dense interstitial tissue. In conclusion, new bone formation regenerated more in experimental than control groups and there was seen observe more regular bony trabecular in experimental than control groups at 4 weeks after surgery. In control group, at 8 weeks after surgery, the defects was filled with scar-like interstitial tissue but, in experimental group, the defects was connected with new bone. Therefore xenograft material had osteoconduction but could not fill the defects. We thought that the effective regeneration of periodontal tissue, could be achieved using GTR with biodegradable membrane.