A Review on the Clinical Characteristics and Outcomes from Spontaneous Intestinal Intramural Hematoma

PURPOSE: Spontaneous intestinal intramural hematoma (SIMH) is a very rare complication of anticoagulation. Most reports on SIMH have been case reports and case series, not well-established clinical studies. Therefore, the aim of this study was to evaluate the clinical characteristics and outcomes from SIMH. METHODS: A retrospective review of the records of 48 patients with non-traumatic SIMH was performed at an urban academic tertiary hospital between January 2001 and December 2012. These patients were diagnosed with SIMH by computed tomography and confirmed by a radiology specialist. Their clinical characteristics and outcomes from SIMH were determined. RESULTS: Among all SIMH cases, the percentage of warfarin users was 70.8%. The median age at presentation was 66.5 years, whereas warfarin users were older (68.0 years) than non-users (55.0 years) (p<0.01). SIMH patients had abdominal pain (81.3%), nausea and vomiting (50.0%) and 62.5% of them had abdominal tenderness. The most frequently involved site was the small bowel (85.4%) and there was only one patient with bowel obstruction from SIMH. A total of 33(68.6%) patients were admitted for 9.3 days for conservative treatment, including transfusion. On the other hand, two patients had surgical intervention. There were no mortality cases from SIMH during the study period. CONCLUSION: SIMH is rare disease which can treated with supportive care. However, it can cause severe complications, such as bowel obstruction and perforation, requiring surgical intervention. Therefore, emergency physicians have to consider SIMH carefully, especially in patients treated with an anticoagulation agent.

The Bacterial Protein Azurin Enhances Sensitivity of Oral Squamous Carcinoma Cells to Anticancer Drugs

PURPOSE: Surgical therapy is the primary treatment for oral cancer, but it can cause facial distortion. Therefore, if anticancer drugs are effective against oral cancer, they may be used preferentially. However, oral squamous carcinoma cells (OSCCs) are resistant to these drugs, so finding a way to enhance the sensitivity of these cells to anticancer drugs is important. The bacterial protein azurin is known to selectively enter cancer cells and induce apoptosis. In this study, we show the anticancer effect of azurin in OSCC. MATERIALS AND METHODS: OSCC cell line (YD-9) was subjected to azurin treatment. Cell viability, morphology and protein expression levels were monitored after treatment of azurin. Cells were also subjected to combination treatment of azurin with either 5-fluorouracil or etopside. RESULTS: Azurin-treated cells showed decreased cell viability accompanied by apoptotic phenotypes including morphological change, DNA breakage, and increases in p53 and cyclin B1 protein levels. Combination treatment of azurin with other anti-tumor agents caused an increase in sensitivity to anticancer drugs in azurin-treated YD-9 cells. CONCLUSION: Azurin has a strong synergistic anticancer effect on oral cancer cells when it is used along with anticancer drugs.

The virulence of Brucella abortus isolated from cattle in Korea / 대한수의학회지

In this study, we isolated 12 of Brucella (B.) spp. from cattle, which have been positive in Rose Bangal test and tube agglutination test in Gyeongbuk province in 2009. According to AMOS PCR analysis, isolated 12 strains were identified as B. abortus. Murine derived macrophage, RAW 264.7 cells, were infected with isolated 12 strains or reference strain (B. abortus 544), and bacterial internalization were characterized. According to these results, we divided the isolated strains into the following three groups: class I, lower internalization than that of B. abortus 544; class II, similar internalization to that of that of B. abortus 544; class III, higher internalization than that of B. abortus 544 within RAW 264.7 cells. Furthermore, intracellular growth, bacterial adherent assay, LAMP-1 colocalization, virulence in mice and surface protein pattern were characterized. From these results, representative strains of class III showed lower LAMP-1 colocalization, higher adherent efficiency, higher virulence in mice than those of B. abortus 544, and showed different pattern of surface proteins. These results suggest that B. abortus field strains, isolated from cattle in Korea, possess various virulence properties and higher internalization ability of field strain may have an important role for its virulence expression.

Dermatologic Diseases Associated with Atopic Dermatitis in Koreans: Multicenter Study / 대한피부과학회지

BACKGROUND: Dermatological diseases can occur with atopic dermatitis. OBJECTIVE: The purpose of this study was to analyze diseases associated with atopic dermatitis in Koreans. METHODS: From November, 2007, to May, 2008, 948 patients with atopic dermatitis who visited the department of dermatology at 19 hospitals were evaluated for associated diseases. RESULTS: Of 948 patients, 53.8% (510) had symptoms associated with other dermatological diseases. In order of frequency, diseases associated with atopic dermatitis included acne, hand/foot eczema, seborrheic dermatitis, urticaria, warts, and recurrent herpes simplex. The number of associated diseases did not change significantly with the severity of atopic dermatitis. However, the incidence of hand/foot eczema and eczema herpeticum correlated significantly with the severity of atopic dermatitis. CONCLUSION: There is a distinct pattern of diseases associated with atopic dermatitis in Koreans.

Recombinant Azurin from Pseudomonas aeruginosa Induces Apoptotic Cell Death in Oral Squamous Carcinoma Cells

The use of bacteria in the treatment of cancer has a long and interesting history. The use of live bacteria in this way however has a number of potential problems including toxicity. Purified low molecular weight bacterial proteins have therefore been tested as anticancer agents to avoid such complications. Oral cancer is a widely occurring disease around the world and these lesions are typically very resistant to anticancer agents. In our present study we investigated the effects of purified recombinant azurin from Pseudomonas (P.) aeruginosa against YD-9 (p53-positive) human oral squamous carcinoma cells. Azurin showed cytotoxic effects against these cells in a dose dependent manner. The cell death accompanied by this treatment was found to be characterized by chromatin condensation and apoptotic bodies. Azurin treatment was further found to increase the expression of p53 The stabilization of p53 and induction of apoptosis in YD-9 cells by azurin suggests that it has potentially very strong anticancer properties in oral squamous carcinoma.

Recombinant Azurin from Pseudomonas aeruginosa Induces Apoptotic Cell Death in Oral Squamous Carcinoma Cells

The use of bacteria in the treatment of cancer has a long and interesting history. The use of live bacteria in this way however has a number of potential problems including toxicity. Purified low molecular weight bacterial proteins have therefore been tested as anticancer agents to avoid such complications. Oral cancer is a widely occurring disease around the world and these lesions are typically very resistant to anticancer agents. In our present study we investigated the effects of purified recombinant azurin from Pseudomonas (P.) aeruginosa against YD-9 (p53-positive) human oral squamous carcinoma cells. Azurin showed cytotoxic effects against these cells in a dose dependent manner. The cell death accompanied by this treatment was found to be characterized by chromatin condensation and apoptotic bodies. Azurin treatment was further found to increase the expression of p53 The stabilization of p53 and induction of apoptosis in YD-9 cells by azurin suggests that it has potentially very strong anticancer properties in oral squamous carcinoma.

Epidemiological Surveillance of P and G Genotypes of Group A Rotavirus Detected from Diarrheic Patients in Daejeon Region

During 3 years surveillance (January 2001 through December 2003) for acute gastroenteritis in human in Daejeon region, 432 out of 4,869 stool samples were selected as rotavirus-positive specimens by means of antigen-capture enzyme-linked immunosorbent assay (ELISA). The P (VP4) and G (VP7) genotypes for 432 stool samples were investigated by reverse transcription polymerase chain reaction (RT-PCR) and nested multiplex PCR. The most prevalent P subtype was P[8] (44.9%), followed by P[4] (25.7%) and P[6] (17.1%). No cases for P[10] and P[9] subtypes were found through the study. In G subtyping, G1 (53.2%) was the most frequently found G type, followed by G2 (23.1%), G3 (9.5%), G4 (6.7%), and G9 (0.9%). The order of detection rates for G2, G3 and G4 was variable by years. The most common G- and P- type combination found in this study was G1P[8] (33.1%), followed by G2P[4] (20.4%), G1P[6] (10.0%), G3P[8] (7.2%) and G4P[6] (4.2%). The mixed types of G and P were observed most frequently in P[8] (1.4%) and G1 (3.2%), respectively. This is the first molecular epidemiological study for Group A rotavirus in Daejeon region. The results might be useful data for evaluating the epidemiological status of rotaviral diarrhea in the region.

Caspases-dependent Apoptosis in Human Melanoma Cell by Eugenol / 대한해부학회지

Malignant melanoma is a highly metastatic tumor, resistant to chemotherapy and radiotherapy. Recent studies have suggested that many therapeutic agents used against cancer mediate their effects by induction of apoptosis of the cancer cells. Eugenol enhances the generation of tissue-damaging free radicals and inflammation or allergic reactions. In particular, it is more cytotoxic against cancer cells compared with normal fibroblasts. This study was performed to investigate whether the cytotoxic effect of eugenol is associated with the induction of apoptosis and involves activation of caspase in the human melanoma G361 cells. Eugenol-induced apoptosis was confirmed by MTT assay, Hemacolor stain, Hoechst stain, DNA electrophoresis, and Western blot analysis. Eugenol had a significant dose- and time-dependent inhibitory effect on the viability of G361 cells. Eugenol treatment induced caspase-3 and -6 cleavage, and activation. The caspase-3 substrates PARP and DFF45 are cleaved during eugenol-induced apoptosis. It was found that the casapase-6 substrate lamin A was cleaved, whose cleavage has been reported to be necessary for complete condesation of DNA during apoptosis. These results suggest that eugenol may constitute a potential antitumor compound against melanoma occurring in the skin and oral mucosa.

Construction of Recombinant DNA with F and HN Genes of Newcastle Disease Virus and Its Immunogenicity

Recombinant DNA vaccines, based on plasmid vectors expressing an antigen under the control of a strong promotor, have several advantages over traditional vaccines. They have been shown to induce a full spectrum of immune responses for humoral and cellular systems and to secure the higher safety and the simplicity of administration. Thus, establishment of DNA vaccines against Newcastle disease virus (NDV) in poultry has been widely investigated using various virus strains and vector systems. In this study, the F and HN genes of NDV CBP-1 strains isolated from diseased pheasants and attenuated by serial passages in egg embryos were cloned using pSLIA vector and constructed two recombinants of pSLIA-tsF and pSLIA-tsHN. The recombinant plasmids were transfected into COS-7 cell and the expression of HN and F proteins were verified by immunofluorescence, SDS-PAGE and Western blot. The recombinant plasmids were injected intramuscularly and intradermally into C57B/6 mouse and a significant increment of HN and F antibodies was detected by ELISA. According to the results, it was implicative that the recombinant DNA could be utilized for development of recombinant DNA vaccine for NDV.

Genetic Characterization of Porcine Circovirus Type 2 Detected from the Pigs in Commercial Swine Farms in Korea

Porcine circovirus type 2 (PCV-2) has been nowadays recognized as a major agent causing postweaning multisystemic wasting syndrome (PMWS) in pigs worldwide. PMWS most commonly affects the weaned piglets, being of increasing importance to the pig industry in Korea. Seven commercial farms affected with PMWS and 2 farms free from PMWS, located in the southern part of Gyeonggi province, were selected for this study. The peripheral mononuclear cells were tested for the presence of ORF2 gene by PCR, and 54 (68.4%) of 79 samples were positive. All of 9 herds tested included the positive cases. The positive rates by herds were 50 to 100% in the PMWS-affected herds and 40 to 62.5% in the PMWS-free herd. The nucleotide and amino acid sequences of ORF2 gene of 6 strains were characterized. Homologies among 6 strains revealed 92.1 to 100% in the nucleotide and 92.3 to 100% in the amino acid. The overall ranges of homologies for 25 strains comprised 2 Korean and 23 foreign strains were 91.1 to 100% in the nucleotide and 89.7 to 100% in the amino acid. Three regions of greater heterogeneity were found in immunorelevant epitopes of the capsid protein, and the sequences between 57 to 80 aa revealed higher mutation than other areas. In the phylogenetic tree analysis, KOR 71 strain was clustered together with Korean strains previously isolated in Korea. The remaining 5 strains were closely clustered with other European and Asian strains. The results will be valuable for improving our understanding of the molecular epidemiology of PCV-2 in Korea and development of preventive measures for PMWS.

Apoptotic Effect of Combination Treatment with a Proteasome Inhibitor, Lactacystin, and a Histone Deacetylase Inhibitor, Trichostatin A, on MCF-7 Cells / 대한해부학회지

Although much information has been accumulated about the synergistic interaction of proteasome inhibitors and HDAC inhibitors to induce apoptosis in a certain type of cells, much less is known currently about the underlying mechanism. This study was undertaken to explore the combination effect of a histone deacetylase inhibitor, TSA, and a proteasome inhibitor, lactacystin, on the induction of apoptosis. Pretreatment of TSA and subsequent treatment of lactacystin showed the strong antitumor activity and nuclear condensation. Western blot assay showed that combination treatment of TSA and lactacystin increased Bax/Bcl-2 ratio and decreased level of XIAP. Activation of caspase-7 and cleavage of PARP were demonstrated after the combination treatment. In combination treatment group, cell cycle arrest was induced at G2/M phase and abolished increase in proteasome activity. This study is elucidating the mechanims whereby targeting apoptotic machineries may help in directing therapeutic strategies.

Detection and Genetic Characterization of Isolates of Hepatitis E Virus from Pigs and Human in Chungnam Region of Korea

Swine hepatitis E virus (HEV) has been reported as a new zoonotic agent due to its close genomic resemblance to the human HEV. Recently this virus is indicated as one of the important pathogens in xenotransplantation that uses pig as a donor animal. We carried out to investigate the prevalence of HEV infections among the pigs and human population in Chungnam region using a nested RT-PCR for detection of a part of HEV ORF2 gene. The sequences of the amplified DNA were analyzed and the genetical divergency were characterized. A total of 18 HEV strains, comprising 16 strains from pig and 2 strains from human, were genetically isolated from the fecal and serum samples. Among the isolates, 5 strains (2.5%) were detected from 200 swine sera and 2 strains (2.0%) from 100 human sera. All of the 16 swine strains were isolated from the pigs at 3 month of age, but none of age groups revealed the positive for swine HEV RNA. In comparison of the nucleotide sequence between 16 swine HEV and 2 human HEV isolates, the range of identities was 91.5% to 100%. Two human HEV isolates shared 99.7% homology. In phylogenetic analysis, all of the isolates were classified into genotype III, and the 18 isolates were also closely related to the prototype of swine HEV and human HEV strains isolated in the United States and others recently identified from swine in Japan and Netherland.

Detection and Genetic Characterization of Isolates of Hepatitis E Virus from Pigs and Human in Chungnam Region of Korea

Swine hepatitis E virus (HEV) has been reported as a new zoonotic agent due to its close genomic resemblance to the human HEV. Recently this virus is indicated as one of the important pathogens in xenotransplantation that uses pig as a donor animal. We carried out to investigate the prevalence of HEV infections among the pigs and human population in Chungnam region using a nested RT-PCR for detection of a part of HEV ORF2 gene. The sequences of the amplified DNA were analyzed and the genetical divergency were characterized. A total of 18 HEV strains, comprising 16 strains from pig and 2 strains from human, were genetically isolated from the fecal and serum samples. Among the isolates, 5 strains (2.5%) were detected from 200 swine sera and 2 strains (2.0%) from 100 human sera. All of the 16 swine strains were isolated from the pigs at 3 month of age, but none of age groups revealed the positive for swine HEV RNA. In comparison of the nucleotide sequence between 16 swine HEV and 2 human HEV isolates, the range of identities was 91.5% to 100%. Two human HEV isolates shared 99.7% homology. In phylogenetic analysis, all of the isolates were classified into genotype III, and the 18 isolates were also closely related to the prototype of swine HEV and human HEV strains isolated in the United States and others recently identified from swine in Japan and Netherland.

Cytoplasmic Domain of Intercellular Adhesion Molecule-1(ICAM-1)is Important for the ICAM-1 Mediated Membrane Projection of Endothelial Cells / 대한해부학회지

Intercellular adhesion molecule-1 (ICAM-1)has been shown to enhance leukocyte adhesion, thereby inducing migration through blood endothelial cells. However, the molecular event during the process of adhesion is largely unknown. To examine the role of ICAM-1 cytoplasmic domain in SDF-1 alpha-induced T lymphocyte migration and adhesion, mutant human ICAM-1 molecules were expressed in COS-7 cell line. COS-7 cells expressing ICAM-1_GFP mutant without alpha-actinin revealed no association with the actin cytoskeleton, while wild-type ICAM-showed clear association with the actin, as observed by confocal microscopy, suggesting that actinin binding motif in the cytoplasmic domain of ICAM-1 is important for the proper localization of ICAM-1 on the cell membrane. However, based on adhesion assay, we found that the cytoplasmic domain of ICAM-1 is not essential for the binding of lymphocytes which were activated by SDF-1alpha. On the other hand, ICAM-1-mediated receptor-ligand clustering event was significantly inhibited in the cells expressing ICAM-1 mutants without alpha-actinin or whole cytoplasmic domain. Taken together, these results suggest that ICAM-1 cytoplasmic domain is not essential for the adhesion but important for the ligand-receptor-mediated membrane projection of endothelial cells before trans-endothelial migration of lymphocytes.

Induction of Apoptosis in Human Oral Squamous Carcinoma Cells by Extracellular Products from Pseudomonas aeruginosa / 대한해부학회지

It was reported that cancer in humans and animals infected with microbial pathogens was regressed about 100 years ago. Bacteria are able to trigger apoptosis by a variety of mechanisms including the secretion of protein synthesis inhibitors, pore forming proteins, molecules activating the endogenous death machinery in the infected cell. This study was conducted in order to investigate whether extracellular products of Psuedomonas aeruginosa (EPPA) induce apoptosis in human oral carcinoma cells (OSC9). The EPPA showed cytotoxic effect on OSC9 cells in dose and time-dependent manner. The cell death was demonstrated to be due to apoptosis characterized by chromatin condensation and nuclear fragment. EPPA treatment induced cleavage of caspase-3 and caspase-6. The caspase substrates, PARP, DFF45 and lamin A were cleaved during EPPA-induced apoptosis. Taken together, EPPA induces apoptosis on human oral squamous carcinoma cells in caspase-dependent manner. Our data therefore provide that EPPA contains a novel antitumor agent for human oral squamous carcinoma.

Cloning and Expression of Hemagglutinin-Neuraminidase Gene of a Thermostable Isolate of Newcastle Disease Virus by Baculovirus Recombinants

The hemagglutinin-neuraminidase (HN) gene of a thermostable Newcastle disease virus isolated from the diseased pheasants in Korea was cloned using Baculovirus transfer vector system, constructing pVL-NDHN inserted with HN gene (1.75 kbp). The HN recombinant baculovirus was generated in Sf-9 cells by co-transfection with pVL-NDHN and linearized baculovirus DNA. The Sf-9 cells infected with the recombinant baculovirus showed the hemagglutinating activity for chicken erythrocytes, and specific positive reactions in indirect immunofluorescence and indirect dot immunoassay. By SDS-PAGE and Western blot analysis, the expressed HN protein with the size of 74 kDa was detected in the cells infected with the recombinant baculovirus. To evaluate the immunogenicity of expressed HN protein, the chicken inoculated with the lysates of the Sf-9 cells were examined by hemagglutination inhibition and ELISA tests. The substantial levels of antibody responses were detected in both assays. The HN protein expressed in baculovirus recombinant system could be utilized for the development of diagnostic measures for Newcastle disease in poultry, and these results on HN recombinant baculovirus will expedite the development of recombinant ND vaccines.

Iron chelator inducesMIP-3alpha/CCL20 in human intestinal epithelial cells: implication for triggeringmucosal adaptive immunity

A previous report by this laboratory demonstrated that bacterial iron chelator (siderophore) triggers inflammatory signals, including the production of CXC chemokine IL-8, in human intestinal epithelial cells (IECs). Microarray-based gene expression profiling revealed that iron chelator also induces macrophage inflammatory protein 3 alpha (MIP-3alpha)/ CC chemokine-ligand 20 (CCL20). As CCL20 is chemotactic for the cells involved in host adaptive immunity, this suggests that iron chelator may stimulate IECs to have the capacity to link mucosal innate and adaptive immunity. The basal medium from iron chelator deferoxamine (DFO)-treated HT-29 monolayers was as chemotactic as recombinant human CCL20 at equivalent concentrations to attract CCR6+ cells. The increase of CCL20 protein secretion appeared to correspond to that of CCL20 mRNA levels, as determined by real-time quantitative RT-PCR. The efficacy of DFO at inducing CCL20 mRNA was also observed in human PBMCs and in THP-1 cells, but not in human umbilical vein endothelial cells. Interestingly, unlike other proinflammatory cytokines, such as TNF-alpha and IL-1beta, a time-dependent experiment revealed that DFO slowly induces CCL20, suggesting a novel mechanism of action. A pharmacologic study also revealed that multiple signaling pathways are differentially involved in CCL20 production by DFO, while some of those pathways are not involved in TNF-alpha-induced CCL20 production. Collectively, these results demonstrate that, in addition to some bacterial products known to induce host adaptive immune responses, direct chelation of host iron by infected bacteria may also contribute to the initiation of host adaptive immunity in the intestinal mucosa.

Soluble glucocorticoid-induced TNF receptor (sGITR) induces inflammation in mice

Glucocorticoid-induced TNF receptor (GITR) was a new member of the TNF/nerve growth factor receptor (TNFR/ NGFR) family and induced in murine T cells by dexamathasone. Recombinant soluble GITR (sGITR) induced an inflammation in peritoneal membrane and changes in spleen after i.p. injection of 3 mg/kg in C57BL/6 mice. Spleen was enlarged and percentage of neutrophils and monocytes were increased. The area of red pulp in spleen was increased, while that of white pulp was decreased after GITR injection. The thickening of membrane and neutrophil infiltration was observed in peritoneal membrane with increased myeloperoxidase activity. At later time, neutrophil infiltration moved to inside the tissue with tissue damage. GITR ligand and GITR were expressed constitutively on the surface of spleen cells and cells from peritoneal fluid. In contrast, no significant change in the spleen and in peritoneal membrane was observed in mice treated with LPS. GITR may play a role in body's inflammatory processes.

Peroneal Nerve Palsy after Total Knee Arthroplasty

PURPOSE: To evaluate prognosis and causative factors of the peroneal nerve palsy that occurred after total knee arthroplasty. MATERIALS AND METHODS: From April 1994 till Feb. 2002, we performed 637 cases(449 patients) of total knee arthroplasty (TKA). Among them we experienced 9 cases(9 patients,1.4%) of peroneal nerve palsy postoperatively. For evaluation of suggestive causes of peroneal nerve palsy, we compared these paralytic group with the control group (628 cases: 440 patients) which has no peroneal nerve palsy after TKA. We analyzed basic patients factors including age, sex, body weight, preoperative diagnosis, and preoperative degree of flexion contracture and tibiofemoral angle, intraoperative tourniquet time, preoperative functional knee scores (HSS and TKSCRS). RESULTS: Eight cases were recovered completely within 12 months after TKA. A case has no change in 1 and 1/2 years after operation so we considered it as a permanent damage. There were no statistical differences between two groups. CONCLUSION: Almost all cases of peroneal nerve palsy were completely recovered spontaneously within 12 months after operation and within 6 months, clinincal sign of improvement was appeared. Until now, we couldn't find any definite causes of peroneal nerve palsy after TKA, so its causes would be multifactorial and further studies may be necessary.

Immunological Responses of Dogs Experimentally Infected with Dirofilaria immitis

Three dogs were experimentally infected with Dirofilaria immitis. All dogs were euthanised at 30, 36 and 37 weeks after inoculation of D. immitis for the recovery of adult worms. Three cases accounted to 42.91 % recovery of inoculated worms. Serum samples from dogs experimentally inoculated with D. immitis were analyzed by ELISA and immunoblotting methods. Antibody titers of dogs detected by ELISA peaked between 7 and 14 weeks then decreased between weeks 15 to 24 followed by another increase during weeks 25 to 30 and persisted throughout the remainde of the experiment period. Analysis of adult D. immitis protein stained with Coomassie brilliant blue R-250 indicated separately more than 10 bands, and the major bands were 22, 40, 46, 56, 70, 72 and 89 kDa. Antigenic identification of extracts antigens of adults D. immitis by immunoblotting analysis revealed several bands from pooled sera of patent infection (30 weeks after inoculation). The detected bands were 24, 70, 80 and 110 kDa, 22, 72 and 84 kDa, and 58 and 72 kDa in dogs 1, 2 and 3, respectively. Results of antibody titers reached high levels on the 4th molting stage after inoculation of infective larva (L3), and reinforced previous findings that high molecular weight regions are detected in young animals.