• Journal of Food Hygiene and Safety
• /
• v.32 no.5
• /
• pp.418-423
• /
• 2017

An analytical method for the determination of dexamethasone (DM) in bovine milk samples was developed and validated using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Milk samples were extracted by the liquid-liquid extraction based on acetonitrile. The chromatographic separation was achieved on a reverse phase $C_{18}$ column with gradient elution using a mobile phase of 0.1% formic acid in 95% acetonitrile. The procedure was validated according to the Ministry of Food and Drug Safety guideline determining accuracy, precision, limit of detection (LOD), and limit of quantification (LOQ). Mean recoveries of DM from spiked milk samples (25, 125, and 1,250 ng/mL) were 98.9-109.6%, and the relative standard deviation was between 1.7 and 4.4%. Linearity in concentration range of 12.5-1,250 ng/mL was obtained with the correlation coefficient ($r^2$) of 0.9997. LOD and LOQ for the investigated DM were 0.15 and 0.5 ng/mL depending on milk samples, respectively. This method was reliable, sensitive, economical and suitable for routine monitoring of DM residues in bovine milk.

• Journal of Life Science
• /
• v.22 no.6
• /
• pp.703-712
• /
• 2012

Adeno-associated virus (AAV) has been considered to be a very safe and efficient gene delivery system. However, the major obstacles to therapeutic usage of AAV have been to achieve highly efficient and reproducible production processes, and also to develop a reliable quantifying method of various serotypes with a simple protocol. We compared the efficiency of the conventional production protocol of AAV2 and adenovirus (Ad) co-infection to that of a new method containing AAV2 infection followed by pHelper transfection. We tested HEK293 and 293T, and further examined the time-dependent changes of AAV2 production. The new method of AAV2 and pHelper DNA gave about ten times higher production efficiency than that of the conventional protocol. The highest production efficiency in 293T was achieved as $1.61{\times}10^5$ virus genomes (v.g.)/cell by the new method of 10 MOI of AAV2 infection and 5 days post-infection. This protocol of the highest efficiency was then applied to produce various AAV serotypes and showed the efficiencies higher than $10^5$ v.g./cell. Next, we designed the universal PCR primers of highly conserved regions for various AAV serotypes to develop a simple and reliable titration method. The universal primers could amplify all the tested AAV serotypes with similar sensitivities by ten molecular copies. Therefore, this pair of universal primers can be further utilized to detect AAV contaminants in therapeutic adenoviral vectors.

• Food Science of Animal Resources
• /
• v.33 no.3
• /
• pp.403-410
• /
• 2013

This study was performed to evaluate the microbiological contamination level of raw beef from retail markets in Seoul, Korea. The sampling and laboratory test were performed according to the procedure of "Standard for processing and ingredients specification of livestock product" and "Korean food code". Enterotoxin of Staphylococcus aureus isolates were detected using VIDAS$^{(R)}$ and PCR-based methods. Listeria monocytogenes serotyping and genotyping were carried out using Listeria antisera and L. monocytogenes Fingerprinting kit, respectively. A total of 48 samples were collected from 16 retail markets (butcher's shop: 5, department store: 6, supermarket: 5) in 2011. The level of total bacteria counts in the butcher's shop, department store and supermarket were $4.4{\times}10^3$ CFU/g, $3.9{\times}10^5$ CFU/g and $1.0{\times}10^4$ CFU/g, respectively. The concentrations of Escherichia coli of these three retail markets were $6.4{\times}10$ CFU/g, 7.6 CFU/g and $2.0{\times}10$ CFU/g, respectively. Salmonella species was not detected on all samples. However, S. aureus was isolated in the 3 samples (6.25%) from each type of three retail markets. L. monocytogenes was isolated in the 4 samples (8.3%) from department stores. The level of contamination of these foodborne bacteria was less than 100 CFU/g. The enterotoxin-encoding genes of S. aureus isolates were sea, seh, sei and sep gene. The gene similarity of L. monocytogenes isolated from two retail markets by Rep-PCR showed 57.8-98.1% and 68.1-98.1%, respectively. These results suggest that the HACCP guideline for environmental control in slaughterhouse and retail markets should be provided to prevent cross contamination and manage foodborne pathogens such as L. monocytogenes and S. aureus.

• Toxicological Research
• /
• v.24 no.1
• /
• pp.29-36
• /
• 2008

The present study investigated the anti-proliferative and chemosensitizing effects of Crinum asiaticum var. japonicum against multi-drug resistant (MDR) cancer cells. The 80% methanol extract, chloroform ($CHCl_3$) fraction and butanol (BuOH) fraction of C. asiaticum inhibited the growth of mitoxantrone (MX) resistant HL-60 (HL-60/MX2) cells. When HL-60/MX2 cells were treated with the $CHCl_3$ and BuOH fractions, DNA ladder and sub-G1 hypodiploid cells were observed. Furthermore, the fractions reduced BcI-2 mRNA levels, whereas Bax mRNA levels were increased. These results suggest that the inhibitory effect of C. asiaticum on the growth of the HL-60/MX2 cells might arise from the induction of apoptosis. Treatment of HL-60/MX2 cells with the fractions markedly decreased the mRNA levels of the multi-drug resistance protein-1 and breast cancer resistance protein. The $CHCl_3$ fraction and hexane fraction increased MX accumulation in HL-60/MX2 cells. These results imply that the $CHCl_3$ fraction of C. asiaticum plays a pivotal role as a chemosensitizer. We suggest that components of C. asiaticum might have a therapeutic potential for the treatment of MDR leukemia.

• Journal of Pharmaceutical Investigation
• /
• v.39 no.6
• /
• pp.401-409
• /
• 2009

Ketrorolac tromethamine (KT), a nonsteroidal anti-inflammatory drug (NSAID) is required repeated administration due to its short blood half-life. To avoid dose-dependent side effects of KT, sustained-release pellets containing KT were prepared by coating with Eudragit$^{(R)}$ RS 100 and Eudragit$^{(R)}$ NE 30D. The in vitro and in vivo drug release behavior of KT from Eudragit$^{(R)}$ RS 100 and NE 30D coated pellet (SR-A), Eudragit$^{(R)}$ RS 100 coated pellet (SR-B) and conventional commercial immediate-release tablet (IR) was investigated. KT from SR-A and SR-B was slowly released over several hours, whereas IR showed rapid initial release in vitro. The pharmacokinetic study in vivo was performed by oral administration in beagle dogs. 5 mg IR was administered 3 times at intervals 5 hr. Five milligrams of IR was administered 3 times at intervals of 5 hr and 15 mg of SR-A and SR-B did once. After administering IR, KT concentration in blood showed high peak- trough fluctuation and stomach ulcer were discovered. On the other hand, SR-A and SR-B sustainedly released KT and reduced the occurrence of stomach ulcer. There sustained-release pellets will be effective system to minimize dosedependent of side effect and improve patient compliance.

• Genomics & Informatics
• /
• v.15 no.3
• /
• pp.87-97
• /
• 2017

Multiple myeloma (MM) is a malignant disease caused by an abnormal proliferation of plasma cells, of which the prognostic factors include chromosomal abnormality, ${\beta}$-2 microglobulin, and albumin. Recently, the term chromothripsis has emerged, which is the massive but highly localized chromosomal rearrangement in response to a one-step catastrophic event. Many studies have shown an association of chromothripsis with the prognosis in several cancers; however, few studies have investigated it in MM. Here, we studied the association between chromothripsis-like patterns and treatment resistance or prognosis. First, we analyzed nine MM cell lines (U266, MM.1S, RPMI8226, KMS-11, KMS-12-BM, KMS-12-PE, KMS-28-BM, KMS-28-PE, and NCI-H929) and bone marrow samples of four patients who were diagnosed with MM by next-generation sequencing-based copy number variation analysis. The frequency of the chromothripsis-like pattern was observed in seven cell lines. We analyzed the treatment-induced chromothripsis-like patterns in KMS-12-BM and KMS-12-PE cells. As a result, breakpoints and chromothripsis-like patterns were increased after drug treatment in the relatively resistant KMS-12-BM. We further analyzed the patients' results according to the therapeutic response, which was divided into sensitive and resistant, as suggested by the International Myeloma Working Group. The chromothripsis-like pattern was more frequently observed in the resistant group. In the sensitive group, the frequency of the chromothripsis-like pattern decreased after treatment, whereas the resistant group showed increased chromothripsis-like patterns after the treatment. These results suggest that the chromothripsis-like pattern is associated with treatment response in MM.

• Journal of Life Science
• /
• v.22 no.8
• /
• pp.991-998
• /
• 2012

Ginseng is one of the most commonly used herbal medicines and health foods. Korean red ginseng (KRG; Panax ginseng C.A. Meyer) extract is known to have potential therapeutic activities, such as anti-viral effects, the amelioration of food allergies, anti-oxidant effects, and obesity reduction. Nevertheless, no reports have been issued the modulatory effects of KRG extract on the activity of cytochrome P450 (CYP). In the present study, we investigated the modulatory effect of KRG extract in vitro and in vivo by using pooled human liver microsomes and male ICR mice. When human liver microsomes were incubated with KRG extract at 0.01-10 mg/ml, CYP1A2, 2B6, 2C19, 2D6, and 3A were not significantly inhibited by KRG extract, although CYP2B6 was slightly inhibited. Mice were orally administered KRG extract at 50, 250, or 500 mg/kg daily for 3, 7, or 14 days. However, the activities of CYPs in mouse livers were not significantly different from those of vehicle-treated controls. In conclusion, no significant ginseng-drug interaction was observed. KRG extract did not significantly modulate the activities of CYPs in vitro or in vivo.

• Korean Journal of Food Science and Technology
• /
• v.37 no.1
• /
• pp.122-128
• /
• 2005

Contamination levels of total aerobic bacteria, coliforms, Esherichia coli, Staphylococcus aureus, Bacillus cereus, and Listeria monocytogenes of Kimbab and its main ingredients sampled from general and specialized restaurants were compared. Total aerobic bacteria and coliform counts of Kimbab samples from both restaurant types were not significantly different (p>0.05), showing approximately $4\;to\;6\;log_{10}CFU/g$. E. coli counts were significantly higher in kimbab from general restaurants ($65%:\;0.7-2.6\;log_{10}CFU/g$) than those from specialized ones ($8.33%:\;0.70log_{10}CFU/g$), whereas those of S. aureus and B. cereus were not significantly different (p>0.05). L. monocytogenes was not detected in all Kimbab samples. These results indicate hygiene of Kimbab and its main ingredients are deleterious. Contamination levels of pathogens determined in the present study may be used as primary data for microbial risk assessment.

• Korean Journal of Veterinary Service
• /
• v.22 no.4
• /
• pp.325-347
• /
• 1999

This study was conducted to investigate the epidemiological, clinicopathological, microbiological, pathological observations and other tests from sudden death in feedlot cattle at the region of Sarari in Korea during the period from 1994 to 1999. Massive or sporadic occurrence of sudden death has been observed in 101 heads of 47 farmhouse. There were 20.8% in spring, 29.7% in summer, 16.8% in autumn, 32.7% in winter, and 62.3% in reproductive, 27.7% in growing, 5.0% in beef cattle, 5.0% in calf in prevalence of sudden death in cattle. Enterotoxemia(88.0%), pneumonia(3.5%), intestinal diarrhea(3.5%), liver abscess(1.5%) and indigestion(1.5%) were detected from 67 heads of sudden death cattle. In clinical observations, cattle were generally died of sudden recumbency with convulsions followed anorexia, depression, ataxia, muscular tremor, tachycardia and dyspnea without any premonitory symptoms. Epidemiological surveys showed no evidence that other factors such as pesticide, insecticide, fertilizer, chemical drug3 and those of others caused sudden death. Macroscopically, there were coagulation disorders of blood, congestion, edema and haemorrhage of lung, congestion and haemorrhages, watery and blood-tinged contents of small intestine. Histopathologically, we observed pulmonary congestion and haemorrhage, necrotic intestinal mucosa accompanied with haemorrhage and congestion, and also increased globule leukocytes between bronchial epithelia with mild pneumonia. Clinicopathologically, only elevation of blood glucose and aspartate aminotransferase(AST) was detected. Magnesium and calcium deficiency were not detected, but parasites were detected highly in normal and dead cattles. Microbiologically, Clostridium(Cl) pefringens were detected from small intestinal contents of 94% (63/67) of sudden death cattle and 51%(51/101) of slaughter cattle, and the population were $10^{6-8}$/cfu/$m\ell$ after 16~32 hours. Consequently, it was proved that the cause of death in cattle was enterotoxemia. Pathogenic test of mouse and goat inoculated with Cl perfringens type A toxin has been demonstrated as similar observation to natural cases. In antimicrobial susceptibility test, ampicillin, bacitracin, polymycin, cephalothin, penicillin, choramphenicol, erythromycin, tetracycline were highly susceptible, and amikacin, gentamicin, kanamycin, neomycin, streptomycin, sulfamethoxine, sulfamethazine were resistant. Cl perfringens were resisted for 4 hours in 3% formalin, 20 minutes in 4% phenol, 20 minutes in 0.5% mercuric chloride and 40 minutes in 0.1% sodium hydroxide, respectively. The useful method to prevent from occurrance of enterotoxemia in feedlot cattle was a dietary administration of antibiotics and miyari acid.

• Korean Journal of Veterinary Service
• /
• v.38 no.3
• /
• pp.173-180
• /
• 2015

Environmental pesticides used for insect control can be transferred from plants to animals even to livestock animals through food chain. Human beings also can be exposed to pesticides by consuming polluted dairy products, including meats, eggs and other milk products. Therefore, the Ministry of Food and Drug Safety (MFDS) established Standard for Pesticide Residue Limits in dairy products. The QuEChERS (quick, easy, cheap, effective, rugged and safe) methods for detecting residual pesticides are relatively well established for fruits and vegetables, however, the methods for meat have not been appropriately studied yet. In the present work, pyraclofos was used as an organophosphate pesticide to examine its tissue residue in experimental animals by QuEChERS methods. For this, pyraclofos (150 mg/kg body weight) was orally administered to male rats once a day for 2 days. After 6, 12, and 24 hr of the treatment, the tissue residues in liver and femoral muscle of the rats were determined using QuEChERS methods followed by HPLC analyses. In preliminary studies, the recovery rates of spiking samples of pyraclofos demonstrated approximately 109~110% from the tissues. In previous study, pyraclofos tissue residues were observed with significantly high levels in livers and muscles at 6 hr of oral treatment. Then, they were almost completely disappeared after 24 hr of the administration, indicating the orally exposed pyraclofos is rapidly absorbed and distributed to body organs, then quickly excreted from the body with a negligible level of tissue residue. The alterations in blood chemistry as well as the histopathology of heart, lung, liver, spleen and kidney have also been investigated in the experimental animals for assessing acute toxic effects of pyraclofos. The obtained blood chemistry indexes (ALT and AST) showed maximum peak values at 12 hr after the oral administration and decreased to the normal levels at 24 hr of the treatment. Histopathologic observation exhibited acute hepatic damages at 24 hr of the treatment. In conclusion, we suggest that QuEChERS method can be adequately optimized for the analysis of pyraclofos residues in animal tissues.