Phenotypic traits, virulence-associated gene profile and genetic relatedness of Edwardsiella tarda isolates from Japanese eel Anguilla japonica in Korea.

UNLABELLED: Edwardsiella tarda is the predominant bacterium in farm-cultured eel in Korea. Here, we evaluated the heterogeneity of 37 E. tarda isolates derived from Japanese eel with various origins (olive flounder, common carp and ornamental fish) between 2003 and 2010. Regardless of origins, the biochemical characteristics of E. tarda isolates were homogenous except hydrogen sulfide production, citrate utilization and mannitol fermentation. Based on the phylogenetic analysis of 16S rRNA, E. tarda isolates could be classified into two subgroups and displayed a close relation with Edwardsiella ictaluri and Edwardsiella hosinae lineages, suggesting that the subgroup I has been a predominant type in the Jeonnam and Jeonbuk provinces. I-CeuI-based pulsed-field gel electrophoresis (PFGE) typing showed that the isolates from Japanese eels belonged to 11 pulsotypes, indicating that the presence of highly genomic diversity. Additionally, two isolates, ET-060 and ET-191, showed a high frequency of virulence genes (100%) and caused 90% and 60% mortality in Japanese eel, respectively. This finding suggests a substantial congruence of virulence gene profiles and pathogenicity. Our results demonstrate that the intraspecific diversity within E. tarda strains from Japanese eel has been in prior existence. SIGNIFICANCE AND IMPACT OF THE STUDY: Based on the biochemical characteristics, the phylogenetic property of the 16S rRNA gene and PFGE types of Edwardsiella tarda, we could identify the intraspecific diversity of isolates from Japanese eel, Anguilla japonica in Korea. In addition, this study describes the strong congruence of virulence-related genes and pathogenicity, suggesting that the virulence profile may be useful tool for prediction of pathogenicity.

Development of a stabilizer for lyophilization of an attenuated duck viral hepatitis vaccine.

The live attenuated vaccine against duck viral hepatitis currently available in Korea requires special freezers for storage and transportation with extra costs involved. The development of a lyophilization stabilizer for live attenuated duck viral hepatitis virus (DHV) vaccines, therefore, has been highly recommended for the wider application of the vaccines. Four conventional vaccine stabilizer formulations containing a disaccharide, such as lactose, trehalose, or sucrose, and new formulations containing sorbitol were tested for their efficacy in stabilizing a new attenuated DHV type 3 vaccine candidate under different storage temperatures, 4 and 37 degrees C. The vaccine virus and each stabilizer formulation were combined and submitted to lyophilization and the viability of the virus was measured in 7-d-old specific-pathogen-free chicken embryos by determining the 50% egg lethal dose. Stabilizer formulations containing 2, 4, or 8% sorbitol preserved the viability of the vaccine virus much better than the other stabilizer formulations and 2% sorbitol was the optimal concentration in a standard stabilizing buffer, phosphate glutamate gelatin (0.0038 M KH2PO4, 0.0071 M K2HPO4, 0.0049 M monosodium L-glutamate, and 0.5% gelatin). The results demonstrate that the stabilizer formulation containing 2% sorbitol and 0.5% gelatin can be used for convenient storage and transportation of live DHV vaccines.

The susceptibility of magpies to a highly pathogenic avian influenza virus subtype H5N1.

Korean wild magpies (Pica pica sericea) were intranasally inoculated with highly pathogenic avian influenza (A/Chicken/Korea/ES/03 virus) (H5N1), which was classified as clade 2.5. We estimated viral replication, death after infection, and histology and immunohistochemistry. This species was highly susceptible to severe infection; 100% of birds died within 5 to 8 d. The virus was detected from oropharyngeal (1 to 5 d postinfection) and cloacal (3 to 5 d postinfection) swabs from infected magpies. At necropsy, the prominent lesions were coalescing necrosis of the pancreas with enlargement of livers and spleens. Microscopically, pancreas, brain, heart, adrenal gland, and kidney were most consistently affected with necrotic and inflammatory changes, and viral antigen was frequently demonstrated in the parenchyma of these organs. As a result, Korean wild magpies were very susceptible to avian influenza (H5N1) virus.

The hydrodynamic filter separator for removal of urban storm runoff.

The majority of storm runoff pollution is trapped in particles smaller than 100 microm in diameter. Solid particles smaller than 100 microm in diameter are not easily separated by conventional types of hydrodynamic separator, and remain in suspension in overflow. To overcome this problem, a HDFS (hydrodynamic filter separator) has been developed for treatment of the microparticles in urban storm runoff. We conducted a laboratory scale study on treatable potential of microparticles using HDFS that combined HDS with perlite filter. To determine the efficiency for various operation conditions, a series of experiments was performed with different solids concentrations and surface loading rate. The operation ranges of surface loading rates were 100 to 2,800 m3/m2/day, and influent solids concentrations were varied from 800 to 1,900 mg/L. Also, the particle size distribution was monitored to investigate the effects of surface loading rates on the particle size. Results indicated that the HDFS-inside type showed greater efficiency at solids separation than the other type.

Highly pathogenic avian influenza in magpies (Pica pica sericea) in South Korea.

Highly pathogenic avian influenza (HPAI) is an extremely infectious, systemic viral disease of birds that produces high mortality and morbidity. HPAI was diagnosed in the three dead magpies (Pica pica sericea) submitted to the National Veterinary Research and Quarantine Service. At necropsy, the prominent lesions were multifocal or coalescing necrosis of the pancreas with enlargement of the livers and spleens. Microscopically, there were severely necrotizing pancreatitis and lymphocytic meningoencephalitis. Influenza viral antigen was also detected in areas closely associated with histologic lesions. Avian influenza virus was isolated from cecal tonsils and feces of the magpies. The isolated virus was identified as a highly pathogenic H5N1, with hemagglutinin proteolytic cleavage site deduced amino acid sequence of QREKRKKR/GLFGAIAG. To determine the pathogenicity of the isolate, eight 6-wk-old specific-pathogen-free chickens were inoculated intravenously with the virus, and all birds died within 24 hr after inoculation. This is the first report of HPAI in magpies.

Comparison of porous bone mineral and biologically active glass in critical-sized defects.

Several materials have been proposed as therapies to augment alveolar bone and to promote periodontal regeneration. However, there are an insufficient number of studies that effectively evaluated these therapies. Consequently, the purpose of this study was to compare bone regeneration promoted by porous bone mineral and biologically active glass. Unilateral critical-sized defects (CSDs) were prepared in the radii of 24 rabbits, divided evenly between 2 time periods (4 and 8 weeks) and between 2 treatment groups (porous bone mineral and biologically active glass). Evaluations consisted of clinical examinations, standardized radiography at baseline and every 2 weeks thereafter, as well as histology and histomorphometry. Data were analyzed by an unpaired Student t-test with significance established at P < or = 0.05. We determined that CSDs treated with porous bone mineral were significantly more radiopaque than biologically active glass-treated sites at both 4 and 8 weeks. Moreover, the amount of new bone was significantly greater at both 4 and 8 weeks in the porous bone mineral groups than in the biologically active glass groups. We concluded that in the rabbit radius CSD wound model, porous bone mineral appears to be more effective than biologically active glass in regenerating bone.

Haemoglobin concentration and oxygen saturation in dog gingiva with experimentally induced periodontitis.

The index of haemoglobin concentration (Hb index) and apparent oxygen saturation (apparent SO2) were determined by a new, non-invasive method, tissue reflectance spectrophotometry. The Hb index was positively correlated with the haemoglobin concentration. The relationship between the apparent SO2 and oxygen partial pressure was a sigmoid curve resembling the haemoglobin oxygen dissociation curve. The Hb index and the apparent SO2 were monitored continuously by tissue reflectance spectrophotometry during the induction of experimental periodontitis with silk ligatures. The Hb index increased rapidly during the first 7 days after ligation and then decreased gradually during the remaining period. The apparent SO2 decreased during the first 7 days but gradually rose during the final 9 weeks. The maximum level of the deoxyhaemoglobin concentration after ligation was elevated about two times over that found before ligation, whereas the increase in oxyhaemoglobin concentration was relatively small. These results suggest that the oxygen supply to inflamed gingiva may increase to some extent, but not sufficiently to compensate for the increased oxygen consumption.

Charged beads enhance cutaneous wound healing in rhesus non-human primates.

Enhanced cutaneous wound healing by positively charged cross-linked diethylaminoethyl dextran beads (CLDD) was studied in a standardized incisional wound model in 20 adult and 20 geriatric Macaca mulatta (rhesus) partitioned equally over five time periods. Physiologic saline served as a control. Soft-tissue linear incisions were prepared between and 1 cm inferior to the scapulae. There were four incisions per rhesus; each incision was 1.5 cm long with 1 cm of undisturbed tissue between incisions, and both the experimental CLDD and physiologic saline treatments were administered to each rhesus. The incision treatments were either CLDD and soft-tissue closure with 4-0 BioSyn sutures or sterile physiologic saline and closure with 4-0 BioSyn sutures. The hypothesis was CLDD would enhance cutaneous wound repair. Verification of the hypothesis consisted of clinical examinations and histologic and tensiometric evaluations on biopsy specimens at 10 and 15 days, whereas 5-day and 2- and 4-month groups were assessed clinically and biopsy specimens were assessed histologically. The clinical course of healing for all groups was unremarkable. At 10 days, incisions in adult rhesus treated with CLDD had a 30-percent greater tensile strength compared with the physiologic saline-treated incisions (p = 0.01), whereas for geriatric rhesus, the CLDD treatment proved to be 15 percent greater in tensile strength compared with the physiologic saline cohort (p = 0.11). By day 15, incisions in adult rhesus were 26 percent stronger than the saline treatment group (p = 0.07), and the difference was 36 percent (p = 0.02) for the geriatric rhesus. From 5 through 15 days, histologic observations revealed a gradual decrease in quantity and integrity of CLDD, with no remnants of CLDD at either 2 or 4 months. Macrophages and multinucleated giant cells were localized in the dermis and were associated with the CLDD. These cells decreased commensurately with the decrease of CLDD beads. The data suggest that CLDD can enhance significantly the tensile properties of healing cutaneous wounds in both adult and geriatric rhesus. Moreover, if the wound healing is enhanced in geriatric patients, this finding may be clinically germane to conditions where wound healing is compromised, such as in diabetics and patients on steroids.

A report of 14,195 applications of anesthetics to oral and maxillofacial surgery at one teaching dental hospital (1971-2000) centering around airway problems.

STUDY OBJECTIVES: To examine the validity of our methods of anesthesia, i.e., awake intubation and assisted manual ventilation, in coping with the anesthetic problems particular to oral and maxillofacial surgery (OMF surgery). DESIGN: Prospective study. SETTING: Operating room and ward of a dental teaching hospital. PATIENTS: 14,195 patients undergoing OMF surgery during the period from January 1971 to March 2000. MEASUREMENTS AND MAIN RESULTS: The kinds of anesthetic difficulties centering around airway problems and their frequency in OMF surgery were determined. In 2,401 patients (16.9%), awake intubation was employed because of definite or possible airway problems. No untoward effects due to awake intubation were noted. Volatile anesthetics were used with nitrous oxide (N2O) in 13,959 patients (98.3%), and their spontaneous respiration were assisted manually for the purpose of early detection of airway troubles such as accidental extubation, dislocation, kinking, and/or damage to the endotracheal tubes. Few accidents or complications were noted in relation to airway issues, and neither cardiac arrest nor death was experienced in these 14,195 patients. CONCLUSIONS: Based on a sufficient number of anesthetic applications, awake intubation and assisted manual ventilation were proved to be useful in coping with the anesthetic difficulties particular to OMF surgery.

The effects of epinephrine and norepinephrine administered during local anesthesia on left ventricular diastolic function.

To evaluate the effects of epinephrine (E) and norepinephrine (NE) administered during local anesthesia on left ventricular diastolic function, we examined transmitral inflow patterns with pulsed Doppler echocardiography. Arterial blood pressure, heart rate, and transmitral inflow patterns were measured in 10 healthy volunteers when 3.6 mL of 2% lidocaine containing 1:80,000 E or 1:25,000 NE was injected supraperiosteally in the maxilla. The dosage of drug administered was 45 micrograms for E and 144 micrograms for NE. After the administration of E, heart rate was increased, but blood pressure was unchanged. Peak early velocity (R), peak atrial velocity (A), the ratio of A to R (A/R), and the deceleration rate (Dc) were increased, whereas the isovolumic relaxation time (IVRT) was shortened. The increase of R and Dc and the shortening of IVRT indicated an activation of left ventricular relaxation. After the administration of NE, blood pressure was increased and heart rate was decreased. A decrease in Dc, shortening of the acceleration half-time (AcT), and prolongation of IVRT were observed. These changes reflect impeded myocardial relaxation. We conclude that E activates left ventricular diastolic function and that NE, in contrast, impairs it.

Recent Korean isolates of duck hepatitis virus reveal the presence of a new geno- and serotype when compared to duck hepatitis virus type 1 type strains.

Duck hepatitis was first reported in 1985 in Korea. The complete nucleotide sequence of two past Korean isolates, DHV-HS and DHV-HSS, isolated in 1994 and 1995, and four recent Korean isolates, AP-03337, AP-04009, AP-04114 and AP-04203 isolated in 2003 and 2004, were determined. Phylogenetic analysis using the 3D protein sequence confirmed that the previously characterized duck hepatitis virus type 1 strains and the six Korean isolates described here constitute a monophyletic group and form two clades/genotypes in which all except the four recent Korean isolates form one group (A) and the recent Korean isolates of 2003 and 2004 constitute a second group (B). Phylogenetic analysis of the VP1 protein supported the division into two different groups. Antisera raised against viruses of group A showed significant neutralizing cross-reaction against a member of the same genotype but not to a strain of group B and vice versa. These results demonstrated that the two genotypes also could be regarded as two different serotypes.

Neural stem cells--a promising potential therapy for brain tumors.

Brain tumors can be highly aggressive and debilitating for many patients and lead to an untimely death in just a few months. Unfortunately, due to the location of many brain tumors, therapy with ionizing radiation, chemotherapeutic agents and/or surgery has limited rewards. In addition, the probability of totally removing highly infiltrative tumors, particularly gliomas, is extremely low and rarely provides a cure. The need for directed targeting and ablation of tumors with minimal damage to nearby healthy tissue has lead to the most recent findings and uses of neural stem cells for therapeutic treatment of brain tumors. Recently, some very promising studies have demonstrated that exogenous neural stem cells have the remarkable ability to migrate very long distances towards sites of metastasis after transplantation. These studies also show that intravascular injections of neural stem cells may lead to preferential migration towards central nervous system tumors. It has also been demonstrated that genetically modified neural stem cells, engineered to produce anti-tumor molecules, upon transplantation, have the ability to migrate towards tumors and reduce tumor mass directly or through a "bystander" effect. Here we review the current literature examining the promise of utilizing genetically modified neural stem cells as vehicles for CNS tumor therapy.

An echocardiographic study of patients with cardiovascular disease during dental treatment using local anesthesia.

The cardiac function of nine patients who had various cardiovascular diseases was investigated by echocardiography during dental treatment in which local anesthetics were administered. The administration of lidocaine with epinephrine enhanced the cardiac output of all patients. Increased heart rate was responsible for the increased cardiac output in four cases, whereas increased stroke volume was responsible in the other five cases. The administration of prilocaine with felypressin caused less of an increase in cardiac function than did lidocaine, with only one patient showing a significant change in cardiac dynamics.