Residual materials such as veterinary drugs, environmental contaminants, and pesticides are affecting food safety. High resolution techniques and quality controls are needed to analyze these materials from part per million to part per trillion quantities in food. In order to achieve quality results, standardized methods and techniques are required. Our laboratories were prepared to obtain a certificate of accreditation for ISO/IEC 17025 in the analytical criteria of animal drugs, dioxins, pesticides, and heavy metals. ISO together with IEC has built a strategic partnership with the World Trade Organization with the common goal of promoting a free and fair global trading system. ISO collaborates with the United Nations Organization and its specialized agencies and commissions, particularly those involved in the harmonization of regulations and public policies including the World Health Organization and CODEX Alimentarius for food safety measurement, management and traceability. Our goal was to have high quality analysts, proper analytical methods, good laboratory facilities, and safety systems within guidelines of ISO/IEC 17025. All staff members took requirement exams. We applied proficiency tests in the analysis of veterinary drugs (nitrofuran metabolites, sulfonamide and tetracyclines), dioxins, organophosphorus pesticides, and heavy metals (Cd, Pb, As) to the Food Analysis Performance Assessment Scheme (FAPAS) at Central Science Laboratory, Department for Environment Food and Rural Affairs (DEFRA), England. The results were very satisfactory. All documents were prepared, including system management, laboratory management, standard operational procedures for testing, reporting, and more. The criteria encompassed the requirements of ISO/IEC 17025:1999. Finally, the Korea Laboratory Accreditation Scheme (KOLAS) accredited our testing laboratories in accordance with the provisions of Article 23 of the National Standards Act. The accreditation will give us the benefit of becoming a regional reference laboratory in Asia.

The primary goal of the wound healing is rapid wound closure. Recent advances in cellular and molecular biology have greatly expanded our understanding of the biologic processes involved in wound repair and tissue regeneration. This study was conducted to develop a new sponge type of biomaterial to be used for either wound dressing or scaffold for tissue engineering. We designed to make a comparative study of the wound healing effect of silk fibroin/hyaluronic acid (SF/HA) blend sponge in full-thickness dermal injury model of rat. Two full-thickness excisions were made on the back of the experimental animals. The excised wound was covered with either the silk fibroin (SF), hyaluronic acid (HA) or SF/HA (7 : 3 or 5 : 5 ratio) blend sponge. On the postoperative days of 3, 7, 10 and 14, the wound area was calculated by image analysis software. Simultaneously, the tissues were stained with Hematoxylin-Eosin and Masson's trichrome methods to measure the area of regenerated epithelium and collagen deposition. In addition, we evaluated the degree of the epithelial cell proliferation using immunohistochemistry for proliferating cell nuclear antigen (PCNA). We found that the half healing time ($HT_{50}$) of SF/HA blend sponge treated groups were significantly decreased as compared with either those of SF or HA treatment group. Furthermore, SF/HA blend sponges significantly increased the size of epithelialization and collagen deposition as well as the number of PCNA positive cells on epidermal basement membrane as compared with those of control treatment. Especially, the 5 : 5 ratio group of SF/HA among all treatment groups was most effective on wound healing rate and histological studies. These results suggest that SF/HA blend sponges could accelerate the wound healing process through the increase of epithelialization, collagen deposition and basal cell proliferation in full thickness skin injury.

Ethanol abuse is associated with liver injury, neurotoxicity, modulation of immune responses, and increased risk for cancer, whereas moderate ethanol consumption exerts protective effects against liver injury. However, the underlying signal transduction mechanisms of insulin-like growth factors (IGFs) which play an important regulatory role in various metabolism mechanisms are not well understood. We investigated the effects of ethanol-induced p42/44 activity on IGF-I secretion, IGF-I receptor and IGFBP-1 secretion using radioimmunoassay and western blotting in primary cultured rat hepatocytes. The p42/44 activity, IGF-I secretion and IGF-I receptor activity significantly accelerated compared to control at 10 and 30 min after 200 mM ethanol treatment, but then it became suppressed at 180 min. In contrast, IGFBP-1 secretion was inhibited compared to control at 30 min after 200 mM ethanol treatment, but increased at 180 min. The IGF-I secretion, IGF-I receptor and p42/44 activity at 30 min after 200 mM ethanol treatment accelerated with increasing ethanol concentration but IGFBP-1 secretion inhibited (p<0.05). The increased IGF-I secretion, inhibited IGFBP-1 secretion and IGF-IR activity by ethanol-induced temporal p42/44 activity at 30 min after ethanol treatment was blocked by treatment with PD98059. Alcohol dehydrogenase (ADH) inhibitor, 4-methylpyramazole blocked the changes of IGF-I secretion, IGFBP-1 secretion, and IGF-IR activity by ethanol-induced p42/44 activity at 30 and 180 min. Taken together, these results suggest that ethanol is involved in the modulation of IGF-I and IGFBP-1 secretion and IGF-IR activity by p42/44 activity in primary cultured rat hepatocytes. In addition, changing of p42/44 activity by ethanol was caused with ADH.

Suaeda (S.) asparagoides $M_{IQ}$, one of the halophyte groups, has been used as a folk remedy for digestive disturbances in Korea. However, its pharmacological activity on gastrointestinal motility has not been reported yet. In this study, the effects of this halophyte extracts with various solvent fractions (ethanol, hexane, chloroform, ethyl acetate, butanol, and water) on mice ileal spontaneous motility was examined. All solvent fractions at the concentration of $100{\mu}g/ml$ showed inhibitory actions on spontaneous motility of ileum with the potency order of water > 70% ethanol > hexane ${\gg}$ chloroform ${\geq}$ butanol ${\geq}$ ethyl acetate, respectively. In addition, the water fraction of extracts from S. asparagoides $M_{IQ}$ (WFSA) dose-dependently ($1-100{\mu}g/ml$) inhibited the amplitude of spontaneous phasic contraction and area under the contractile curve (AUC). The inhibitory effect of water fraction at the concentration of $10{\mu}g/ml$ was not affected by tetrodotoxin (TTX), $Na^+$ channel blocker ($1{\mu}M$), and $N^w$-nitro-L-arginine Methyl Ester (L-NAME), nitric oxide synthase inhibitor ($100{\mu}M$). However, cyclopiazonic acid (CPA, $10{\mu}M$), inhibitor of sarcoplasmic reticulum $Ca^{2+}$-ATPase, almost blocked the inhibitory effects of WFSA ($10{\mu}g/ml$) on the spontaneous phasic contraction of mouse ileum. But, CPA did not inhibit the lowering basal tone effects of WFSA. The result of this study showed that various extracts of S. asparagoides $M_{IQ}$ induce inhibitory effects on spontaneous contraction of mice ileal segments. More over, the polar solvent fractions were shown to be more potent than non-polar solvent fractions. The effects of S. asparagoides $M_{IQ}$ extracts are not mediated by nerve or nitric oxide. The inhibitory effects of WFSA at least partially mediated by sarcoplasmic reticulum $Ca^{2+}$-ATPase. However, further study is required to determine the exact pharmacological mechanisms of this halophyte on its gastrointestinal motility inhibitory effects.

A stable and reliable Helicobacter pylori (H. pylori) infection animal model would be necessary for evaluating vaccine efficacy and helpful for understanding the pathological mechanism of the organism. The aim of the present study is to investigate the effect of ethanol treatment prior to H. pylori inoculation on associated gastric mucosal injury and to establish ethanol-pretreating animal model to study H. pylori infection. Male Mongolian gerbils were used for the study. H. pylori was orally inoculated after 12 h fasting. 3 h prior to H. pylori inoculation, a group of gerbils was orally treated with absolute ethanol, 60% and 40% ethanol respectively. Another group of animals was treated either with H. pylori culture media alone or with different concentrations of ethanol plus culture media. Gerbils were killed 4 or 8 weeks after H. pylori inoculation. The colonization of H. pylori was confirmed by both histological examination and rapid urease test. Mucosal damage was evaluated grossly and histologically according to the criteria. The colonization of H. pylori and pathological changes in gastric mucosa of the animals were also observed. Although no significant change to the gastric mucose was observed in the animals treated either with H. pylori culture media alone or with different concentrations of ethanol plus culture media, persistent H. pylori infection was seen in the mucosa and mucosal leucocyte infiltration and severe epithelial damage was observed in the Helicobacter and ethanol + Helicobacter groups after 4 weeks. The gross and histological scores were higher in the ethanol + Helicobacter than in the Helicobacter alone group. As the results, ethanol-pretreatment with 60% concentration induced severe pathogenic changes by H. pylori infection in 5 weeks-old Mongolian gerbils. These results suggested that ethanol-pretreatment before H. pylori inoculation could increase the severity of gastric mucosal inflammation and enhance the colonization of H. pylori. The established ethanol-pretreating animal model would contribute to screen new drugs against H. pylori and be used as an useful tool for various animal experiments with H. pylori strains.

The shigellae are common etiological agents of bacillary dysentery in humans and primates. During four years from 2002 to 2005, 22 strains of Shigella spp. were isolated from the diarrheic patients in Seoul region. All of them were identified as S. flexneri by biochemical tests and serotyping. The prevalence of serotypes were variable by year, but the major serotypes were 2a and 3a. In an antimicrobial susceptibility test, all of the isolates were resistant to streptomycin and tetracycline, and susceptible to amikacin, kanamycin, cefoxitin, and gentamicin. All of the isolates showed the multi-resistant patterns over 3 drugs. By analysis of the plasmid profile the isolates were classified into 7 groups (P1~P7). Serotypes 2a and 2b were distributed to P1, P2, P3, and P4. Serotype 3a was differentiated to P5 and serotype 3b, to P6 and serotype 4a, to P7. PCR results showed that all isolates were positive for two virulence genes, ipaH and ial, but none of the strains had stx gene. The set1A and set1B genes were detected from 12 isolates (54.5%) that belonged to serotype 2a and 2b. The sen gene was detected from 19 isolates (86.4%). The 22 isolates showed 12 to 17 DNA fragments in the sizes ranging from 20.5 kb to 1135 kb, resulting in 13 patterns by the PFGE with Not I digestion. The PFGE patterns of the isolates showed the close relation with the serotypes, but no relations with year of isolation and antimicrobial resistance.

Serum samples of 30 chickens per flock from 6 grand parent stock (GPS) farms and 70 parent stock (PS) farms were collected for seroprevalent study of pullorum disease-fowl typhoid (PD-FT) infection by serum plate agglutination test (SPA). The incidence of PD-FT infection in GPS flocks and PS flocks were 0% and 15.7%, respectively. Especially PS flocks infected with PD-FT showed age dependent patterns that 22.2% of flocks between 20 to 30 weeks of age and 38.9% of flocks between 30 to 40 weeks of age were positive. The incidence of GPS flocks and PS flocks using Salmonella (S.) gallinarum 9R (SG9R) live vaccine were 33.3% and 58.6%, respectively. The sero-positive rate of 11 flocks were 6.7-83.3% by SPA and 2.9-55.6% by enzyme linked immunosorbent assay (ELISA), and ELISA showed more lower antibody levels than SPA. Furthermore, specific antibodies produced by SG9R vaccination were detectable by SPA using SG9R antigen without cross-reaction with the PD-FT infection.

Porcine epidemic diarrhea virus (PEDV) strain Chinju99, which was previously isolated from piglets suffering from severe diarrhea was used to characterize the membrane (M) protein gene to establish the molecular information, and the results will be useful in elucidating concepts related to molecular pathogenesis and antigenic structures of PEDV isolates. The Chinju99 M gene generated by reverse transcription and polymerase chain reaction (RT-PCR) consisted of 681 bases containing 22.3% adenine, 22.3% cytosine, 23.1% guanine and 32.3% thymine nucleotides, and the GC content was 45.4%. It had some nucleotide mismatches from M gene of other PEDV strains, such as CV777, Br1/87, KPEDV-9, JMe2, JS2004-2 and LJB-03 with 97-99% nucleotide sequence homology to these strains. Also, it encoded a protein of 226 amino acids, which had some mismatches from those of CV777, Br1/87, KPEDV-9, JMe2, JS20004-2 and LJB-03, as the amino acid sequence homology showed a 97-98% to these strains. The Chinju99 had a very close relationship to the Japanese strain JMe2 for the nucleotide and amino acid sequences of the M gene. The amino acids predicted from Chinju99 M gene consisted of mostly hydrophobic residues and contained three potential sites for asparagine (N)-linked glycosylation, two serine (S)-linked phosphorylation sites by protein kinase C, and two S- or threonine (T)-linked phosphorylation sites by casein kinase II.

Porcine reproductive and respiratory syndrome (PRRS) is an economically important disease of swine that occurs all over the swine industry worldwide. It was first observed in the Unite States in 1987 then in Europe in 1990. It has been described in Japan and in Korea in 1993. PRRS virus is divided into two distinct types, North American and European, genetically. Based on our limited knowledge there has been no report on the existence of European PRRSV. But according to the government's Korea Customs Service there has been many importations of breeding pigs from Europe. These seem to make an estimate that European PRRSV could be introduced in Korea by inflow of European breeding pigs. We first detected the European PRRSV could be introduced in Korean pig farms by using polymerase chain reaction (PCR). Further, it is also identified that there are not only North American PRRSV antibody but also a European PRRSV antibody. According to the genetical and serological experiment results, the presence of established North American PRRSV in Korea is due to the use of live vaccines made of North American PRRSV strain as well field virus infection, and the European PRRSV is possibly introduced from imported breeding stock.

Newcastle disease virus (NDV) is a single-stranded negative sense RNA virus, which has been classified as a member of the Avulavirus genus of the Paramyxoviridae family. It is also one of the most important pathogens in the poultry industry. The glycoproteins, fusion (F) and hemagglutinin-neuraminidase (HN), determine the virulence of NDV, and the relevant molecular structures have already been determined. NDV isolates differ in terms of virulence, and at least 2 of 9 genotypes (I-IX) have been shown to co-circulate. Therefore, it is clearly important to differentiate between vaccine strains and field isolates. In vivo pathogenicity tests have been the standard protocol for some time, but molecular methods appear preferable in terms of the rapidity of diagnosis, as well as animal welfare concerns. In this study, we have designed primer sets from HN gene for phylogenetic analysis and restriction enzyme analysis, and from F gene for pathotype-specific RT-PCR. Via the combination of 2 methods, 106 Korean NDV isolates obtained from 1980 to 2005 were differentiated into vaccine strains, and virulent genotypes VI and VII. The genotype VI viruses were only rarely isolated after 1999, and genotype VII, after it was initially isolated from poultry in 1995, recurred in 2000, and then became the main NDV constituting a threat to the Korean poultry industry.

Discospondylitis is an infection of intervertebral discs, associated end plates, and adjacent vertebral bodies causing destruction and proliferation. A 10-year-old intact female Rhodesian Ridgeback dog was referred to the Veterinary Medical Teaching Hospital, Seoul National University with paraparesis and severe pain at T13-L1 region for one week duration. The treatment with PDS for one week by referring veterinarian showed no clinical improvement. At time of presentation, clinical signs of the patient included depression, vaginal discharge, and fever. In T-L spine radiography, osteolytic change was found between caudal endplate of T13 and cranial endplate of L1. The affected vertebral bodies were shortened and intervertebral disc space was widened. In abdominal radiography, a dilated soft tissue opacity tubular structure was found in mid-caudal abdomen. The abdominal ultrasonography revealed the uterus dilated with echogenic fluid. The CT findings showed concentric lysis of T13-L1 endplates. The dog was diagnosed as diskospondylitis at T13-L1 and pyometra. Urine and blood were cultured, and Staphylococcus (S.) intermedius was identified. The patient had surgical treatment for pyometra but died for septicemia.

Progressing to the iron deficiency anemia was experimentally induced in 4 clinically healthy dogs by repeated phlebotomy to characterize hematologic features, serum iron values, and RBC indices. Abnormal RBC morphologies were also evaluated semiquantitatively on Wright's-stained blood films. Hematologic abnormalities in early stage of anemia included decreased both hematocrit and hemoglobin, and reticulocytosis, with no changes in mean corpuscular volume (MCV) and mean corpuscular hemoglobin concentration (MCHC) were represented. In intermediate stage, decreased serum iron concentration with microcytosis and hypochromia were prominent. In late stage, red cell distribution width and Mentzer's index were out of reference ranges in the majority of dogs. In this study microcytic anemia was appeared at the hemoglobin range of 5.1-7.2 g/dl. On most sampling days, platelet counts and white blood cells were within the reference ranges, with some minor variations. Iron deficiency was not necessarily associated with microcytic anemia. Judging from the sequential changes of both MCV and MCHC, 3 patterns of anemia were sequentially observed: initially normocytic normochromic, intermediate normocytic hypochromic or normocytic normochormic, and finally microcytic hypochromic. The most frequent morphologic abnormalities were target cells. Occasional elliptocyte, acanthocyte, stomatocyte, kinzocyte, dacrocyte and schistocyte were also noted on the blood films.

A 2-month-old female Shih-tzu dog was referred because of lameness, exercise intolerance, depression, elbow and stifle joint swelling. Physical examination, complete blood counts, serum-chemistry, radiography, synovial fluid analysis, antinuclear antibody test, and rheumatoid factor measurement were initiated. On radiography, soft tissue swelling of elbow and stifle joints without erosiveness were founded. The results of synovial fluid analysis revealed severe neutrophilic pleocytosis (nondegenerative), decreased viscosity, increased turbidity, positive on mucin-clot test, and negative on bacterial culture. The results of rheumatoid factor measurement and antinuclear antibody test were negative and below 1 : 40, respectively. Based on all tests, we diagnosed this case as juvenile onset type I immune-mediated polyarthritis. Azathioprine (1 mg/kg body weight, per os q 24 h, for 4 weeks) was then administered and clinical signs improved gradually. Four weeks after azathioprine administration, clinical signs were disappeared. This report describes the clinical findings, imaging characteristics, synovial fluid findings, and other laboratory results of type I immune-mediated polyarthritis and successful management with azathioprine therapy.

Caudal occipital malformation syndrome (COMS) is relatively common disease which occurs at craniocervical junction. It has been known that this syndrome was developed mainly in Cavalier King Charles spaniels, but these days there are a lot of case reports about COMS in other small breed dogs. In this report, COMS was diagnosed in ten dogs presented to Haemaru Referral Animal Hospital using MRI examination from January 2005 to April 2006. These cases were also investigated about concurrent diseases and syringomyelia was the most common type of complication, but clinical signs were nonspecific. COMS could be controlled by medical therapy with corticosteroid except cases with concurrent meningitis. It is considered that the present report could provide information regarding imaging and clinical features of COMS such as concurrent diseases, clinical signs, and prognosis in small animal practice despite of limited case numbers.

A 4-year-old male Miniature Pinscher was referred because of head tilt, nystagmus, and ataxia. The hemogram revealed a moderate neutrophilic leukocytosis. On magnetic resonance imaging, cerebellar inflammation was detected. And cerebrospinal fluid analysis indicated marked neutrophilic pleocytosis. Based on these results, bacterial meningoencephalitis was suspected. The clinical signs were well controlled by a combination antibiotics therapy of the third generation cephalosporins (cefotaxime and cefixime) and metronidazole. We tentatively diagnosed this case as a bacterial meningoencephalitis because clinical signs were improved after only antibiotics therapy and relapsed when stopped antibiotics administrations, even though the result of bacterial culture on communication of cerebrospinal fluid (CSF) was negative.