• Asian-Australasian Journal of Animal Sciences
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• 제18권10호
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• pp.1498-1504
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• 2005

Competitive direct ELISA was developed to detect gentamicin residues. Mice immunized with gentamicin-keyhole limpet hemocyanin (KLH) conjugate developed good antiserum titers, which gradually increased with booster injections, indicating immunization was successfully processed. Monoclonal antibody against gentamicin was prepared using hybridoma cells cloned by limit dilution of fused cells. IgG was purified from ascites fluid of hybridoma cell-injected mice through ammonium sulfate precipitation and Sephadex G-25 gel filtration. After the gel filtration, fractions of high antibody titer were further purified through affinity chromatography on protein A/G column. Monoclonal antibody against gentamicin was confirmed as IgG1, which has kappa light chain. Cross-reactivities ($CR_{50}$) of gentamicin monoclonal antibody to other aminoglycosides (kanamycin, neomycin, and streptomycin) were less than 0.005%, indicating the monoclonal antibody was highly specific for gentamicin. Standard curve constructed through competitive direct ELISA showed measurement range (from 80 to 20% of B/$B_0$ ratio) of gentamicin was between 1 and 40 ng/ml, and 50% of B/$B_0$ ratio was about 4 ng/ml. The gentamicin concentration rapidly increased to 1,300 ng/ml after the intramuscular administration up to 2 h, then sharply decreased to less than 300 ng/ml after 4 h of withdrawal, during which the elimination half-life ($t_{1/2}$) of gentamicin in the rabbit plasma was estimated to be 1.8 h. Competitive direct ELISA method developed in this study using the prepared monoclonal antibody is highly sensitive for gentamicin, and could be useful for detecting gentamicin residues in plasma of live animals.

• 한국임상수의학회지
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• 제17권2호
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• pp.327-333
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• 2000

The diagnostic method was, evaluated in experimentally induced acute renal failure in doges. Ten male dogs weighing from 5 to 10 kg were assigned to two groups(gentamicin & ethylene glycol treated group) al random. Gentamicin sulfate at 10 mg/kg of body weight, t.i.d., for 7 days and ethylene glycol at 3 ml/kg of body weight once used in a randomized complete block design with tee treatments in block. The samples(blood, urine) were collected before and 1,3,5,7,9 and l1th day after administration. The serum creatinine concentration and BUN(blood urea nitrogen) were sig- nificantly increased at the 7th day than before administration in gentamicin treated group (p<0.05), bolt there was significant increase at the 1st day than before administration in ethylene glycol treated group(p<0.05). The urine GGT(gramma glutamyl transpeptidase) and GGT/creatinine were significantly increased at the 1st (lay after administration in gentamicin treated group (p<0.05). But in ethylene glycol treated group, there was no significant changes. The value of FENa (fractional excretion of sodium) was significantly increased at the 3rd day after administration in gentamicin treated group and the 1 st day after administration in ethylene glycol treated group (p<0.05). These results suggest that FENa was a good parameter of renal function in dogs with acute renal failure.

• Macromolecular Research
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• 제8권6호
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• pp.253-260
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• 2000

Biodegradable wafers were prepared with poly (hydroxybutyrate-co-hydroxyvalerate) (PHBV;5, 10, and 15 mole% for 3-hydroxyvalerate) by simple heat pressing method for the sustained release of antibiotic agent, gentamicin sulfate (GS) to investigate the possibility of the treatment for osteomyelitis. The effects of hydroxyvalerate (HV) content, thickness of wafers, various types of additives such as sodium dodecyl sulfate (SDS), microcrystalline cellulose, polyvinylpyrrolidone, and hydroxypropylcellulose (HPC), and different initial drug loading ratio on the release profile have been investigated. In vitro release studies showed that different release patterns and rates could be achieved by simply modifying factors in the preparation conditions. PHBV wafers with 3 mm thickness, 10% of GS initial loading, 15% of HV content and addition of 5% of SDS and HPC were free from initial burst and a near-zero-order sustained release was observed for over 30 days. It might be suggested that the mechanisms of G5 release may be more predominant simple dissolution and diffusion of GS than erosion of PHBV in our system.

• 폴리머
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• 제25권3호
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• pp.318-326
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• 2001

본 연구에서는 세포 배양시 감염방지용으로 널리 쓰이는 겐타마이신 설페이트(GS)가 서방화된 다공성 PLLA 지지체를 유화동결건조방법으로 제조하였다 이들의 물성을 전자현미경 및 수은다공도계로 특성결정하였고, 적심성은 푸른염색 수용액으로 관찰하였으며, 방출거동은 high performance liquid chromatography (HPLC)를 이용하여 측정하였다. GS가 5, 10 및 20%가 포접된 PLLA 지지체의 다공도는 80~90%이었으며 평균다공크기 범위는 30~57 ${\mu}{\textrm}{m}$ 그리고 가장 큰 것으로는 150${\mu}{\textrm}{m}$이상의 것도 관찰되었다. 전체적인 다공도의 모양은 다공과 다공사이의 연결이 양호하고 대부분이 열린 셀 구조를 하고 있는 것으로 나타났다. 대조군에 비해 GS의 농도가 증가함에 따라 PLLA 지지체의 다공도가 감소하는 것으로 보아 GS내의 설페이트 부분은 친수성 역할 그리고 겐타마이신 부분은 소수성 역할을 수행하는 계면활성제의 역할에 기인한 것으로 사료된다. PLLA 지지체의 방출거동은 GS의 농도가 증가함에 따라 방출되는 양이 증가하였고, 적심성 또한 향상되어 세포배양시 긍정적 효과를 끼칠 것으로 예상되었다.

• 한국식품위생안전성학회지
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• 제9권3호
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• pp.123-131
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• 1994

An enzyme-linked immunosorbent assay(ELISA) was developed for the detection of residual gentamicin(GM) in edible animal products. The immunogen(GM-KLH conjugate) and coating antigen(GM-BSA conjugate) were prepared by coupling GM sulfate to keyhole limpet hemocyanin(KLH) and bovine serum albumin(BSA) in the presence of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, respectively. Polyclonal antibody to GM was produced in rabbits(New Zealand White, female) by using the immunogen and the antibody titer was measured by indirect ELISA. A competitive ELISA was developed using GM-bovine serum albumin conjugate as a coating antigen, GM(as standards or sample), polyclonal antibody to GM, secondary antibody conjugated with horseradish peroxidase as an enzyme, and H2O2 and o-phenylenediamine dihydrochloride as a substrate and a chromophore, respectively. The detection limit of GM was 10 ng/ml and the standard curve of GM(n=26) was linear up to 10 $\mu\textrm{g}$/ml in this competitive ELISA system. There were no cross-reactivities of the partially purified antibody between GM and the various antibiotice such as amikacin, benzyl-penicillin, chloramphenicol, erythromycin, furazlidone, kanamycin, neomycin, oleandomycin, streptomycin, sulfathiazole and thiamphenicol(CR50<0.05%)

• 한국세라믹학회지
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• 제56권1호
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• pp.56-64
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• 2019

Calcium phosphate bone cement was prepared to contain antibiotics for release after setting using granulated ${\beta}$-tricalcium phosphate (${\beta}$-TCP) and hydroxyapatite (HA). Gentamicin sulfate (GS) solution was infiltrated within the interconnected pores of the granule to avoid affecting the setting reaction and for protection of GS during the setting. Consequently, the setting time and the temperature increase were not affected, regardless of the loading of GS. The release of the GS from the cement was estimated by measuring the concentration at regular intervals from the cement dipped solution. The ${\beta}$-TCP granule loaded with GS exhibited the saturation of accumulative concentration at 16 h. In contrast, the HA granule with GS exhibited steady increase in accumulative concentration of over $10{\mu}g/ml$ at 144 h. Thus, the granulated cement could release the GS greater than the minimum inhibitory concentration of staphylococcus during the prescription peroid of the oral antibiotics.

• 대한수의학회지
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• 제29권3호
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• pp.393-405
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• 1989

This study was carried out to treatment test for bovine mastitis by the determination of adenosine triphosphate (ATP) based on firefly bioluminescence. The results obtained are followed; 1. In the susceptibility test, cephalothin which looks the most effective were sensitive to Staphylococcus sp. (72.3%), Micrococcus sp. (84.2%), Streptococcus sp. (72.7%) and Gram positive bacilli (72.7%), Gram negative bacilli were sensitive to gentamicin (92.3%) and Yeast-like-fungi was the most sensitive to clotrimazole, and nystatin in order. 2. When the number of bacteria, such as Staphylococcus aureus, Candida tropicalis isolated from the mastitis milk were counted by conventional agar plating technique, and compared with the concentration of bacterial ATP, it gave a good linear relationship. The content of ATP per Staphylococcus aureus, cell was 3.1fM and Candida tropicalis showed the high level of A TP (90fM). 3. The ATP assay was applied to the determination of minimal inhibitory concentration (MIC) of various antibiotics. When Staphylococcus aureus was incubated in the presence of different concentration of tetracycline, erythromycin, kanamycin and streptomycin sulfate and the growth was monitored by the conventional agar plating technique and ATP assay, both methods shown the same results that they were 1mcg/ml, 2mcg/ml, 6.25mcg/ml and 8mcg/ml, respectively. 4. For the determination of susceptibility of sensitive and resistant Staphylococcus au reus isolated for the milk with mastitis to tetracycline, erythromycin kanamycin and streptomycin sulfate, the minimum time required for the test was determined by the assay of ATP every 30 minutes during incubation of 3 hours at $37^{\circ}C$. ATP concentration time curve calculated on both resistant and sensitive strains incubated 3 hours as the optimum time for the determination of susceptibilities of various antibiotics exemed. The ATP concentration of each test broth (antibiotic containing), expressed as a percentage of its own control brith (antibioticfree) indicated values of 30% to be indicative of each antibiotic sensitivity. Single time point ATP assay carried out on the various sensitive and resistant of Staphylococcus aureus to antibiotics examined after 3 hours at $37^{\circ}C$ correlated exactly with disc diffusion and MIC. 5. In the cure of intramammary treatment of bovine mastitis in lactating quarters, the cure rate of Staphylococcal mastitis showed to cephalexin (80%), cloxacillin and gentamicin (70%), ampicillin and oxytetracycline (60%), and Streptococcal mastitis showed to cephalexin (85%), penicillin (80%), cloxacillin and oxytetracycline (75%), and ampicillin (70%), but intramammary antimycotic drug (clotrimazol) were only a little effect about fungal mastitis.

• 대한수의학회지
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• 제30권2호
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• pp.203-214
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• 1990

A total of 166 strains of Salmonella (S) typhimurium var copenhagen isolated from pigeons (164 strains) and aquatic birds (2 strains) were examined for the biochemical characteristics and plasmid profiles. All the strains were sensitive to ampicillin, chloramphenicol, gentamicin, kanamycin and sulfadimethoxine. But 13 strains(7.8%) were resistant to streptomycin (Sm), 2 (1.2%) to tetracycline, 2 (1.2%) to rifampicin, and 1 (0.6%) to nalidixic acid. Among drug resistant strains, only one strain resistant to Sm contained conjugative R plasmid which was fertility inhibition and incompatibility group $I_{\alpha}$. All the strains were sensitive to cobalt chloride, cupric sulfate, lead nitrate, mercuric chloride and silver nitrate. Of 166 isolates, 6 (3.6%) were resistant to sodium arsenate and 1 (0.6%) to potassium tellurite. Among 166 isolates, 1 (0.6%) was colicinogenic, 12 (7.2%) sucrose fermenters, and 166 (100%) maltose fermenters. Plasmid profiles were confirmed as being 4 or 5 plasmids, and their molecular weight ranged 3.2 to 60 megadalton (MD). All the strains harbored 60 Md plasmid. There are three patterns by the plasmid profile, 150 isolates (90.4%) were pattern I (3.2, 3.5, 33, 60Md), 14 (8.4%) pattern II (3.2, 3.5, 29, 60Md), and 2 (1.2%) pattern III (4.2, 7.8, 8.5, 15, 60Md). S typhimurium var copenhagen strains containing 60Md plasmid were resistant to killing by 90% normal guinea pig serum.

• Journal of Periodontal and Implant Science
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• 제30권2호
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• pp.257-277
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• 2000

New techniques for regenerating the destructed periodontal tissue have been studied for many years. Current acceptable methods of promoting periodontal regeneration are basis of removal of diseased soft tissue, root treatment, guided tissue regeneration, graft materials, and biological mediators. Platelet Rich Plasma has been reported as a biological mediator which regulates activities of wound healing progress including cell proliferation, migration, and metabolism. The purpose of this study is to evaluate the effects of using the Platelet Rich Plasma as a regeneration promoting agent for furcation involvement defect. Five adult beagle dogs were used in this experiment. The dogs were anesthetized with Ketamin HCl(0.1 ml/kg, IV)and Xylazine hydrochloride($Rompun^{(R)}$, Bayer, 0.1 ml/kg, IM) and conventional periodontal prophylaxis were performed with ultrasonic scaler and hand instruments. With intrasulcular and crestal incision, mucoperiosteal flap was elevated. Following decortication with 1/2 high speed round bur, degree II furcation defect was made on mandibular third(P3), forth(P4) and fifth(P5) premolar, and stopping was inserted. After 4 weeks, stopping was removed, and bone graft was performed. Ca-P was grafted in P3(experimental group I), Combination of Ca-P and plasma rich platelet were grafted in P4(experimental group II), and P5 was remained at control group.Systemic antibiotics(gentamicin sulfate)and anlgesics(phenyl butazone) were administrated intramuscular for 2 weeks after surgery. Irrigation with 0.1% Chlorhexidine Gluconate around operate sites was performed during the whole experimental period except one day immediate after surgery. Soft diets were fed through the whole experiment period. After 4, 8 weeks, the animals were sacrificed by perfusion technique. Tissue block was excised including the tooth and prepared for light microscope with Gomori's trichrome staining. At 4 weeks after surgery, there were rapid osteogenesis phenomenon on the defected area of the Platelet Rich Plasma plus Ca-P BBP group and early trabeculation pattern was made with new osteoid tissue produced by activated osteoblast. Bone formation was almost completed to the fornix of furcation by 8 weeks after surgery. In conclusion, Platelet Rich Plasma can promote rapid osteogenesis during healing of periodontalregeneration.

• Macromolecular Research
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• 제11권4호
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• pp.207-223
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• 2003

For last five years, we are developing the novel local drug delivery devices using biodegradable polymers, especially polylactide (PLA) and poly(D,L-lactide-co-glycolide) (PLGA) due to its relatively good biocompatibility, easily controlled biodegradability, good processability and only FDA approved synthetic degradable polymers. The relationship between various kinds of drug [water soluble small molecule drugs: gentamicin sulfate (GS), fentanyl citrate (FC), BCNU, azidothymidine (AZT), pamidronate (ADP), $1,25(OH)_2$ vitamin $D_3$, water insoluble small molecule drugs: fentanyl, ipriflavone (IP) and nifedipine, and water soluble large peptide molecule drug: nerve growth factor (NGF), and Japanese encephalitis virus (JEV)], different types of geometrical devices [microspheres (MSs), microcapsule, nanoparticle, wafers, pellet, beads, multiple-layered beads, implants, fiber, scaffolds, and films], and pharmacological activity are proposed and discussed for the application of pharmaceutics and tissue engineering. Also, local drug delivery devices proposed in this work are introduced in view of preparation method, drug release behavior, biocompatibility, pharmacological effect, and animal studies. In conclusion, we can control the drug release profiles varying with the preparation, formulation and geometrical parameters. Moreover, any types of drug were successfully applicable to achieve linear sustained release from short period ($1{\sim}3$ days) to long period (over 2 months). It is very important to design a suitable formulation for the wanting period of bioactive molecules loaded in biodegradable polymers for the local delivery of drug. The drug release is affected by many factors such as hydrophilicity of drug, electric charge of drug, drug loading amount, polymer molecular weight, the monomer composition, the size of implants, the applied fabrication techniques, and so on. It is well known that the commercialization of new drug needs a lot of cost of money (average: over 10 million US dollar per one drug) and time (average: above 9 years) whereas the development of DDS and high effective generic drug might be need relatively low investment with a short time period. Also, one core technology of DDS can be applicable to many drugs for the market needs. From these reasons, the DDS research on potent generic drugs might be suitable for less risk and high return.