Thrombin Induced Apoptosis through Calcium-Mediated Activation of Cytosolic Phospholipase A2 in Intestinal Myofibroblasts

Thrombin is a serine protease that participates in a variety of biological signaling through protease-activated receptors. Intestinal myofibroblasts play central roles in maintaining intestinal homeostasis. In this study, we found that thrombin-induced apoptosis is mediated by the calcium-mediated activation of cytosolic phospholipase A2 in the CCD-18Co cell. Thrombin reduced cell viability by inducing apoptosis and proteinase-activated receptor-1 antagonist attenuated thrombin-induced cell death. Endogenous ceramide did not affect the cell viability itself, but a ceramide-mediated pathway was involved in thrombin-induced cell death. Thrombin increased intracellular calcium levels and cytosolic phospholipase A2 activity. The ceramide synthase inhibitor Fumonisin B 1, intracellular calcium chelator BAPTA-AM, and cytosolic phospholipase A2 inhibitor AACOCF 3 inhibited thrombin-induced cell death. Thrombin stimulated arachidonic acid release and reactive oxygen species generation, which was blocked by AACOCF 3, BAPTA-AM, and the antioxidant reagent Trolox. Taken together, thrombin triggered apoptosis through calcium-mediated activation of cytosolic phospholipase A2 in intestinal myofibroblasts.

Correlations in the results of virus neutralization test, hemagglutination inhibition test, and enzyme-linked immunosorbent assay to determine infectious bronchitis virus vaccine potency

The virus neutralization (VN) test was used to determine potency of the infectious bronchitis (IB) vaccine. The results of VN, hemagglutination inhibition (HI), and enzyme-linked immunosorbent assay (ELISA) were compared with those of the IBV M41. The r² values between VN and HI titers and the ELISA antibody titer were 0.8782 and 0.0336, respectively, indicating a high correlation between VN and HI, but not VN and ELISA. The Cohen's kappa coefficient between the VN titer of 2 log₁₀ and HI titer of 5 log₂ was 0.909. Our results showed that VN could be replaced with HI for testing the potency of IBV M41.

Preparation and diagnostic utility of a hemagglutination inhibition test antigen derived from the baculovirus-expressed hemagglutinin-neuraminidase protein gene of Newcastle disease virus

A recombinant hemagglutinin-neuraminidase (rHN) protein from Newcastle disease virus (NDV) with hemagglutination (HA) activity was expressed in Spodoptera frugiperda cells using a baculovirus expression system. The rHN protein extracted from infected cells was used as an antigen in a hemagglutination inhibition (HI) test for the detection and titration of NDV-specific antibodies present in chicken sera. The rHN antigen produced high HA titers of 2(13) per 25 microL, which were similar to those of the NDV antigen produced using chicken eggs, and it remained stable without significant loss of the HA activity for at least 12 weeks at 4degrees C. The rHN-based HI assay specifically detected NDV antibodies, but not the sera of other avian pathogens, with a specificity and sensitivity of 100% and 98.0%, respectively, in known positive and negative chicken sera (n = 430). Compared with an NDV-based HI assay, the rHN-based HI assay had a relative sensitivity and specificity of 96.1% and 95.5%, respectively, when applied to field chicken sera. The HI titers of the rHN-based HI assay were highly correlated with those in an NDV-based HI assay (r = 0.927). Overall, these results indicate that rHN protein provides a useful alternative to NDV antigen in HI assays.

Non-Adherence to Psychiatric Intervention in Work Related Burns

PURPOSE: Patients with work related burns suffer from anxiety, depression, insomnia and suicide ideation etc. Psychiatric symptoms could be reduced by treatment. Almost all patients are referred to psychiatric intervention in our hospital. However, a number of patients show non-adherence. The aim of this study is to figure out the reason of non-adherence and psychiatric symptoms of work related burns patients. METHODS: 123 patients participated in this study. Startle, Physiological arousal, Anger, and Numbness (SPAN), Feeling Suicide, Patient Health Questionnaire-2 (PHQ-2) were administered as screening tool. Questions about psychiatric intervention and reasons to refuse psychiatric intervention were asked. RESULTS: 32% patients were depressed, 34% patients had suicide ideation and 59% had PTSD after work related burns. However, 46% of burn patients had not been treated. Stigma of psychiatric intervention and concerns about dependency were major reasons for non-adherence. CONCLUSION: There are gap between necessity and reality of psychiatric intervention on work related burn patients.

Generation and Biological Characterization of a Neutralization-Resistant Mutant of Newcastle Disease Virus

A neutralization-resistant mutant of Newcastle disease virus (NDV) Kr005 strain belonging to class II genotype VII was generated using a neutralizing monoclonal antibody and its biological effects were assessed. The mutant showed single amino acid substitution (E to K) at position 347 of the hemagglutinin-neuraminidase (HN) protein (E347K mutant). The E347K mutant exhibited marked rounding of the cells and few syncytia in infected chicken embryofibroblast (CEF) cells. The hemadsorption and neuraminidase activities of the E347K mutant of the wild-type virus were 118% and 166%, respectively. The mutant produced a rapid elution pattern whereas the wild type had a slow elution pattern. Growth kinetics studies showed that the E347K mutant produced an 80-times higher yield of extracellular virus in CEF cells compared with the wild-type virus. The time-course virus titer showed a marked increase in mutant-infected cells from 6 h to 12 h post infection (pi), which was consistent with the titer pattern time-course for NA activity. The E347K mutant virus showed a slight decrease in virulence compared to the wild-type virus, but there was no change in pathotype when measured by in vivo pathogenicity testing. These results suggest that an E347K mutation in HN protein might be associated with increased NA activity and subsequent enhancement of virus release from infected cells without change in viral pathotype.

Development of an attenuated vaccine strain from a korean respiratory type infectious bronchitis virus / 대한수의학회지

An attenuated vaccine strain AVR1/08 of Korean respiratory type of infectious bronchitis virus (IBV) was developed by 89th passages of IBV D85/06 strain in chicken eggs. The AVR1/08 strain had higher virus titer at least 20 times (10(1.3)) than the parent virus D85/06 by egg inoculation method. The AVR1/08 strain had a single point mutation (S to Y) at position 56 of spike protein of IBV compared to parent virus IBV D85/06 strain. The mutation was observed consistently at viruses after 47th passage in chicken eggs. The AVR1/08 strain showed no virulence even after 6 passages in chickens and all chickens inoculated induced anti-IBV antibody 14 days after vaccination. The AVR1/08 strain had broad protective efficacy against QX type Korean nephropathogenic virus (Q43/06 strain), KM91 type Korean nephropathogenic virus (KM91 strain) and Korean respiratory virus (D85/06 strain). In contrast, Massachusetts (Mass) type attenuated vaccine strain H120 showed protection of 37.5 to 50% against these three viruses. Our results indicate that the AVR1/08 strain has potential as an attenuated vaccine effective in controlling IBVs circulating in Korea.

Development of Competitive ELISA for Detection of Avian Metapneumovirus Antibodies in Chicken

Avian metapneumovirus (aMPV) causes an acute and highly contagious upper respiratory tract infection in turkeys and chickens. In this study, a competitive ELISA (C-ELISA) was developed for the detection of antibodies to aMPV in chicken sera and/or their egg yolks. This assay is based on the competitive binding of monoclonal antibody with serum antibodies to recombinant aMPV N protein expressed by a recombinant baculovirus. The C-ELISA showed specificity and sensitivity of 100% and 98.0%, respectively, when compared to the virus neutralization test. In specific pathogen-free chickens experimentally infected with aMPV SC1509 strain, the C-ELISA started to detect antibodies to aMPV as early as 5 days post infection from birds infected with aMPV, while a commercial ELISA kit detected first 10 days post infection. The C-ELISA was similar or superior to a commercial ELISA kit when serum and egg yolk samples collected from chickens on six outbreak farms were tested for diagnosis. The C-ELISA developed in the present work provides a short turnaround time and can be a useful diagnostic and screening tool for aMPV infection in the field.

Isolation and Characterization of Avian Metapneumovirus from Broiler Breeder Chickens in Korea

Avian metapneumovirus (AMPV) is an emerging pathogen causing respiratory and reproductive illness in poultry worldwide. To demonstrate the presence of AMPV in domestic chickens in Korea, we attempted to isolate AMPV from affected chickens. A cytopathic agent was isolated using chicken tracheal ring culture from dead chickens from a broiler breeder farm with reduced egg production in Korea. This agent, termed SC1509 strain, subsequently passed in Vero cells with distinct cytopathic effects. The SC1509 strain was confirmed as avian metapneumovirus (AMPV) using both RT-PCR test and monoclonal antibody-based immunofluorescence assay. Sequence analysis based on the G glycoprotein revealed that the SC1509 strain had 22.5 to 96.0% nucleotide sequence identity and 11.1 to 92.7% predicted amino acid sequence identity with previously published AMPV strains, particularly with the highest sequence homology (95.8 to 96% for nucleotides and 92.2 to 92.7% for amino acids) to European strains belonging to genotype B. The SC1509 strain was phylogenetically clustered with genotype B viruses, confirming that the SC1509 strain belongs to genotype B. This is the first report of genotype B avian metapneumovirus from chickens in Korea.

Characterization of Lentogenic Newcastle Disease Virus Isolated in Jeju, Korea during 2007~2008 Surveillance

To expand the epidemiological understanding of Newcastle disease in Jeju Province, Korea, active surveillance was extensively performed through a virological examination for poultry farms and wild birds in Jeju Province during 2007~2008. Samples (swabs or fresh feces) were collected from a total of 6,485 birds including 6,405 domestic birds (chickens, ducks, pheasants, geese, quails, turkeys, and ostriches) and 80 wild birds. A total of 24 hemagglutinating agents were isolated from domestic birds on fourteen farms including five Korean native chicken, one layer chicken, two broiler chicken, four duck and two pheasant farms. The hemagglutinating agents were all identified as lentogenic NDV based on the reverse transcriptase polymerase chain reaction, sequence analysis of amino acids on the F cleavage site and mean death time in chicken embryos. The F gene-based phylogenetic analysis revealed that the NDV isolates were classified into genotypes 1 or 2 of class II. These lentogenic viruses were closely related to NDV vaccine strains used in Jeju Province. Active surveillance conducted for Newcastle disease indicates no scientific evidence of virulent NDV infection in chickens in Jeju Province, Korea since 2005.

Pathogenicity and antigenicity of a new variant of Korean nephropathogenic infectious bronchitis virus

Despite the existence of an active vaccination program, recently emerged strains of nephropathogenic infectious bronchitis virus (IBV) in Korea have caused significant economic losses in the poultry industry. In this study, we assessed the pathogenic and antigenic characteristics of a K-IIb type field strain of IBV that emerged in Korea since 2003, such as Kr/Q43/06. Specific pathogen free 1-week-old chickens exhibited severe respiratory symptoms (dyspnea) and nephropathogenic lesions (swollen kidneys with nephritis and urate deposits) following challenge with the recent IBV field strain. The antigenic relatedness (R value), based on a calculated virus neutralization index, of the K-IIb type field strain and K-IIa type strain KM91 (isolated in 1991) was 30%, which indicated that the recent strain, Kr/Q43/06, is a new variant that is antigenically distinct from strain KM91. This report is the first to document the emergence of a new antigenic variant of nephropathogenic IBV in chicken from Korea.

Detection of Infectious Bursal Disease Virus by Double Antibody Sandwich ELISA

Infectious bursal disease virus (IBDV) is responsible for a highly contagious disease of poultry causing severe immunosuppression in chickens. A double antibody sandwich ELISA (DAS-ELISA) was developed to detect IBDV from clinical samples. Two kinds of anti-IBDV antibodies, monoclonal antibody R63 and chicken anti-IBDV sera, were used for DAS-ELISA. Detection limit of IBDV by DAS-ELISA was approximately 10(2.7) EID(50)/ml. The DAS-ELISA detected IBDV from most (13/14) of vaccine products including mild, intermediate and intermediate-plus types. The DAS-ELISA also detected IBDV from all (19/19) of field Korean isolates including very virulent and intermediate-plus phenotypes. Our results indicate that the DAS-ELISA would provide useful diagnostic tool to detect IBDV from clinical samples as well as rapid quantitative detection of IBDV.

Detection of Antibodies to Infectious Bursal Disease Virus (IBDV) by Agar Gel Immunodiffusion using Recombinant VP2 Protein

Infectious bursal disease virus (IBDV) causes a highly contagious and immunosuppressive disease of chicken. Agar gel immunodiffusion using IBDV antigen extracted from bursa of Fabricius of infected chicken has been used officially for diagnosis of IBDV in Korea. In this study, in order to replace the IBDV whole virus antigen with non-infectious antigen, recombinant VP2 protein (rVP2) of IBDV was produced using recombinant baculovirus expression system. Purified baculovirus-expressed rVP2 was used as an antigen in an agar gel immunodiffusion (AGID). rVP2 antigen precipitated specifically IBDV antibodies. AGID using rVP2 antigen detected anti-IBDV antibodies from 6 dpi to 28 dpi (termination of the experiment) when specific pathogen free chickens were experimentally infected with IBDV 52/70 strain. This was consistent with result by AGID using IBDV antigen, virus neutralization test (VNT) and a commercial ELISA kit (except for one serum). The sensitivity of rVP2 was the same with that of IBDV antigen when field sera (n=324) were tested by AGID. However, AGID using rVP2 antigen detected maternal antibodies from broiler chickens (n=20) on a broiler farm up to 15 days old, although the detection rate of the AGID was relatively low compared to a commercial ELISA kit. Our results indicate that IBDV whole virus antigen from IBDV infected chickens would be replaced with recombinant VP2 protein as an antigen for AGID.

Comparison of Oral Hygiene Effects between 0.1% Chlorhexidine and Normal Saline on the Incidence of Oral Pathogens / 기본간호학회지

PURPOSE: The purpose of this study was to compare the effects of oral hygiene with 0.1% chlorhexidine or with normal saline on the incidence of pathogens in the oral cavity of patients in Intensive Care Units (ICU). METHOD: A quasi experimental design with non-equivalent control group and non-synchronized design was used. For the study 46 patients were recruited from a university hospital (24 for the experimental group, 22 for the control group). patients in the experimental group received mouth care with 0.1% chlorhexidine gluconate and those in the control group received mouth care with normal saline twice a day for 7 days in a row. Oral samples were taken for bacterial cultures on admission day, the 4th day and the 7th day for both groups. RESULTS: The incidence of oral pathogens decreased in the experimental group, and increased in the control group. There was no significant difference in the incidence of oral pathogens between the two groups. However oral hygiene using 0.1% chlorhexidine gluconate decreased the incidence of oral pathogens significantly for patients who already had pathogenic bacteria in their mouths on the admission day. CONCLUSION: The results suggest that mouth care with 0.1% chlorhexidine is effective for decreasing the incidence of oral infection for ICU patients who have oral infections.

Morphological changes in rat testes after Hindlimb Unloading model / 항공우주의학회지

BACKGROUND: The purpose of this study was to examine histological changes of testes induced by hindlimb unloading & position change of testis in mature adult male rats. METHOD: Eight month old (390~410g) 36 male Sprague-Dawley rats were weight-matched assigned to Ground Control (GC), Hindlimb Unloading (HU), Hindlimb Unloading & Testis Tie(HUT), Intra-abdominal Testis(IAT) for 24 days. Daily body weight was watched for status of animal. Testis weight, Seminiferous Tubule Diameter(STD), Seminiferous Tubule Perimeter(STP), Serminiferous Tubule Area(STA), Sertori Cell Height(SCH) were examined in each group. RESULTS: The 24-day hindlimb unloading of HU, HUT and IAT showed no significant change of body weight compared to GC. HUT maintained the weights, STD, STP, STA, SCH of their testes as GC while HU, IAT had significantly decreased the weights, STD, STP, STA of their testes. CONCLUSION: These results support that hindlimb unloading experiment induces serious morphological changes of testes. Therefore, on the hindlimb unloading experiment, IAT should be avoid or the effect of IAT should be correct. It will be necessary to study the pure effect of hindlimb unloading without IAT on the musculoskeltal system and so on which are associated with testosterone.

The Modulation of Radiosensitivity by Combined Treatment of Selective COX-2 Inhibitor, NS 398 and EGF Receptor Blocker AG 1478 in HeLa Cell Line / 대한방사선종양학회지

PURPOSE: Selective inhibition of multiple molecular targets may improve the antitumor activity of radiation. Two specific inhibitors of selective cyclooxygenase-2 (COX-2) and epidermal growth factor receptor (EGFR) were combined with radiation on the HeLa cell line. To investigate cooperative mechanism with selective COX-2 inhibitor and EGFR blocker, in vitro experiments were done. MATERIASL AND METHODS: Antitumor effect was obtained by growth inhibition and apoptosis analysis by annexin V-Flous method. Radiation modulation effects were determined by the clonogenic cell survival assay. Surviving fractions at 2 Gy (SF2) and dose enhancement ratio at a surviving fraction of 0.25 were evaluated. To investigate the mechanism of the modulation of radiosensitivity, the cell cycle analyses were done by flow cytometry. The bcl-2 and bax expressions were analyzed by western blot. RESULTS: A cooperative effect were observed on the apoptosis of the HeLa cell line when combination of the two drugs, AG 1478 and NS 398 with radiation at the lowest doses, apoptosis of 22.70% compare with combination of the one drug with radiation, apoptosis of 8.49 %. In cell cycle analysis, accumulation of cell on G0/G1 phase and decrement of S phase fraction was observed from 24 hours to 72 hours after treatment with radiation, AG 1478 and NS 398. The combination of NS 398 and AG 1478 enhanced radiosensitivity in a concentration-dependent manner in HeLa cells with dose enhancement ratios of 3.00 and SF2 of 0.12 but the combination of one drug with radiation was not enhanced radiosensitivity with dose enhancement ratios of 1.12 and SF2 of 0.68 (p=0.005). The expression levels of bcl-2 and bax were reduced when combined with AG 1478 and NS 398. CONCLUSION: Our results indicate that the selective COX-2 inhibitor and EGFR blocker combined with radiation have potential additive or cooperative effects on radiation treatment and may act through various mechanisms including direct inhibition of tumor cell proliferation, suppression of tumor cell cycle progression and inhibition of anti-apoptotic proteins.

Case Report of Osteochondroma on the Mandible Body Area

Osteochondroma is a relatively common benign tumor of skeleton, occurring most often in the metaphyseal region of long bone. However, it is very rare in the facial bone, expecially in the mandible body. We report a case of osteochondroma on the left mandible body in a 9-years-old male, which has been present for 2 years. It was successfully removed by surgery. On pathologic examination, it was diagnosed as osteochondroma consisting of bony structures covered with a cartilaginous cap, the typical composition of osteochondroma.

Bleeding from a Tuberculous Gastric Ulcer in a Hemodialysis Patient / 대한신장학회잡지

The incidence of Tuberculosis among the patients with end-stage renal disease(ESRD) has increased up to 16 times of that in the general population. The impairment of the cellular immunity in the ESRD patients may have a role in the pathogenesis. Extrapulmonary tuberculous manifestations such as lymph node, peritoneum, and pleura involvement are more frequent in the ESRD patients than in the general population. However, there has been no case of upper gastrointestinal(UGI) bleeding as a result of a gastric tuberculosis in the ESRD patient on hemodialysis. Here we report an unusual case of a hemodialysis patient with UGI bleeding secondary to a tuberculous gastric ulcer. A 31-year-old female on hemodialysis was admitted with melena. Endoscopy revealed a benign gastric ulcer with a visible bleeding vessel at the base, located in the anterior wall of the antrum. An exploratory laparotomy showed multiple, round, small and yellow nodules on the visceral peritoneum as well as a 1-cm sized gastric ulcer. After gastric resection, a histological examination including peritoneal nodules demonstrated chronic granulomatous inflamation with caseous necrosis and giant cells. The patient has been on antituberculosis medication and followed up in the outpatient department without any event for 8 months.

Various Clinical Applications of Human Dermal Allograft(AlloDerm(R))

No abstract available.

The Effect of Corticosteroid Therpy in the Very Premature Infant

PURPOSE: Our purpose was to deterrnine the efficacy of maternal corticosteroid therapy on the morbidity of premature infants between 26 and 31 weeks' gestation. METHOD: A total of 62 premature infants between 26 and 31 weeks gestation admitted to pediatric department of National Medical Center from Nov, 1990 to June 1996 were analyzed to evaluate the efficacy of prenatal corticosteroid therapy on the morbidity such as hyaline membrane disease, intreventricular hernorrhage, necrotizing enterocolitis, sepsis, neonatal death, days on ventilation and hospital days. RESULT: Among 62 women who delivered premature infants between 26 and 31 weeks, 22 received betamethasone before delivery and 40 did not. 1) The rate of hyaline membrane disease was less in the betamethasone group (41% vs. 70%, P<0.05). 2) The rate of intracranial hemorrhage was less in the betamethasone group (0 vs. 20%, P<0.05). 3) The days of ventilator care was less in the betamethasone group among survival cases (27+/-3.2 vs. 5.2+/-4.6, P<05). 4) Arnong 14 women who delivered at 26 to 28 weeks, 4 received betamethasone before delivery and 10 did not. The rate of neonatal death was less in the betamethasone group (o% vs. 80%, P<0.05). 5) Among 48 women who delivered at 29 to 31 weeks, 18 received betamethasone before delivery and 30 did not. The rate of hyaline membrane disease was less in the betamethasone group (39% vs. 73%, P<0.05). CONCLUSION: Betamethasone appears to reduce hyaline membrane disease, intraven- tricular hemorrhage, neonatal death and the morbidity significantly in premature infants between 26 and 31 weeks' gestation.

The Change of Serum Cholesterol level in Children with Fever

PURPOSE: This study was undertaken to assess the change of the serum cholesterol level according to the duration of fever in children. METHODS: A retrospective study presents data on fasting serum cholesterol assessed 346 children aged from 3 to 14 years, admitted to National Medical Center from Jan. 1992 to Jun. 1994 due to febrile disease. These patients were divided into three groups according to the duration of fever at the time of cholesterol measurement. Group 1 comprised children in whom cholesterol was evaluated on the first 2day of fever, group 2 children in whom cholesterol was assessed on day 3 to 5 and group 3 subjects evaluated 6 to 20 days after the onset of fever. 200 healthy children without fever were also enrolled in this study. The fasting serum cholesterol levels were also measured in both febrile period and afebrile convalescent period in 34 children with febrile diseases during the 7months period of Apr. to Oct. 1994. RESULTS: As the retrospective study, the fasting serum cholesterol levels of group 1, 2 and 3 were lower than those of the controls. And the difference of the serum cholesterol levels between group 1 and group 3 was significant. The prospective study was significantly associated with a decrease of serum cholesterol in the febrile period compared with that in the convalescent period. CONCLUSIONS: Timing of blood collection for screening test and diagnostic measurement of serum cholesterol level should be carefully defined taking into account that febrile illnesses are biologic variables that may significantly affect serum cholesterol.