Comparative proteomic analysis of outer membrane protein 43 (omp43)-deficient Bartonella henselae

Outer membrane proteins (OMPs) of Gram-negative bacteria constitute the first line of defense protecting cells against environmental stresses including chemical, biophysical, and biological attacks. Although the 43-kDa OMP (OMP43) is major porin protein among Bartonella henselae-derived OMPs, its function remains unreported. In this study, OMP43-deficient mutant B. henselae (Δomp43) was generated to investigate OMP43 function. Interestingly, Δomp43 exhibited weaker proliferative ability than that of wild-type (WT) B. henselae. To study the differences in proteomic expression between WT and Δomp43, two-dimensional gel electrophoresis-based proteomic analysis was performed. Based on Clusters of Orthologus Groups functional assignments, 12 proteins were associated with metabolism, 7 proteins associated with information storage and processing, and 3 proteins associated with cellular processing and signaling. By semi-quantitative reverse transcriptase polymerase chain reaction, increases in tldD, efp, ntrX, pdhA, purB, and ATPA mRNA expression and decreases in Rho and yfeA mRNA expression were confirmed in Δomp43. In conclusion, this is the first report showing that a loss of OMP43 expression in B. henselae leads to retarded proliferation. Furthermore, our proteomic data provide useful information for the further investigation of mechanisms related to the growth of B. henselae.

In Vitro Activity of Diphenyleneiodonium toward Multidrug-Resistant Helicobacter pylori Strains

BACKGROUND/AIMS: The increased resistance of Helicobacter pylori to antibiotics has increased the need to develop new treatments for this bacterium. The aim of our study was to identify new drugs with anti-H. pylori activity. METHODS: We screened a small molecule library—the library of pharmacologically active compounds (LOPAC), which includes 1,280 pharmacologically active compounds—to identify inhibitors of H. pylori growth. The minimal inhibitory concentrations (MICs) of antibiotics against multidrug-resistant H. pylori strains were determined using the agar dilution method. RESULTS: We identified diphenyleneiodonium (DPI) as a novel anti-H. pylori agent. The MIC values for DPI were <0.03 μg/mL against all tested H. pylori strains. DPI also exhibited strong antibacterial activity against common gram-negative and gram-positive pathogenic bacteria. CONCLUSIONS: DPI may be a candidate anti-H. pylori drug for future development.

Synchronous Peripancreatic Lymph Node Gastrinoma and Gastric Neuroendocrine Tumor Type 2

A 34-year-old man was referred to our hospital with gastric polypoid lesions and biopsy-confirmed neuroendocrine tumor (NET). Computed tomography (CT) revealed a 3×3.5×8-cm retroperitoneal mass behind the pancreas, with multiple hepatic metastases. His serum gastrin level was elevated to 1,396 pg/mL. We performed a wedge resection of the stomach, a right hemi-hepatectomy, and a retroperitoneal mass excision. After careful review of the clinical, radiological, histopathological, and immunohistochemical findings, peripancreatic gastrinoma, and synchronous gastric NET were ultimately diagnosed. We reviewed a CT scan that had been performed 6 years previously after surgery for a duodenal perforation. There was no evidence of gastric or hepatic lesions, but the retroperitoneal mass was present at the same site. Had gastrinoma been detected earlier, our patient could have been cured using less invasive treatment. This case demonstrates how important it is to consider Zollinger-Ellison syndrome in patients with a recurrent or aggressive ulcer.

Expression and Characterization of Genes by Expressed Sequence Tag Analysis in the Rat Thymus during Regeneration following Acute Thymic Involution Induced by Cyclophosphamide / 체질인류학회지

The thymus is the central lymphoid organ for the development of bone marrow-derived precursor cells into mature T-cells. Understanding the molecular mechanism of thymic involution and regeneration is critical to develop methods to normalize or improve host immunity from the decreased immune function caused by thymic involution. In this study, the regenerating thymus cDNA library was constructed in the rat from a model of thymic involution and regeneration induced by cyclophosphamide. Expressed sequence tags (ESTs) were obtained by partial sequencing of 700 randomly selected insert-containing clones. A total of 630 ESTs were analyzed, of which 486 ESTs (78%) matched to known genes and 125 ESTs (19%) matched to other ESTs (unknown genes). The 19 ESTs (3%) did not match with any known sequences. The ESTs were grouped into six main functional categories: metabolism (44%), signaling components (20%), membrane transport (7%), cytoskeleton (2%), cell division (2%) and defense (2%). As a result of RT-PCR analysis, expression of putative gene 01, putative E2IG2 gene, musculin and osteoactivin significantly increased in rat thymus during regeneration. The putative gene 01 showed complete homology with mitochondrial ribosomal protein S4 by homology search and multiple alignment of amino acid. These results provide the extensive molecular information on thymus regeneration and will be useful source to identify various genes which may play an important role in the thymus regeneration as well as to clone novel genes. Furthermore, the availability of these data will serve as a basis for further research to understand the molecular mechanism of thymus regeneration.

Enhanced proliferation and differentiation of Oct4- and Sox2-overexpressing human adipose tissue mesenchymal stem cells

Mesenchymal stem cells (MSCs) are attractive candidates for clinical repair or regeneration of damaged tissues. Oct4 and Sox2, which are essential transcription factors for pluripotency and self-renewal, are naturally expressed in MSCs at low levels in early passages, and their levels gradually decrease as the passage number increases. Therefore, to improve MSC proliferation and stemness, we introduced human Oct4 and Sox2 for conferring higher expansion and differentiation capabilities. The Oct4-IRES-Sox2 vector was transfected into human adipose tissue MSCs (ATMSCs) by liposomal transfection and used directly. Oct4 and Sox2 were successfully transfected into ATMSCs, and we confirmed maintenance of MSC surface markers without alterations in both red fluorescent protein (RFP) (control) and Oct4/Sox2-ATMSCs. Enhanced proliferative activity of Oct4/Sox2-ATMSCs was shown by WST-1 assay, and this result was further confirmed by cell counting using trypan blue exclusion for a long period. In addition, FACs cell cycle analysis showed that there was a reduction in the fraction of Oct4/Sox2-ATMSCs in G1 with a concomitant increase in the fraction of cells in S, compared with RFP-ATMSCs. Increased levels of cyclin D1 were also seen in Oct4/Sox2-ATMSCs, indicating acceleration in the transition of cells from G1 to S phase. Furthermore, Oct4/Sox2-overexpressing ATMSCs showed higher differentiation abilities for adipocytes or osteoblasts than controls. The markers of adipogenic or osteogenic differentiation were also upregulated by Oct4/Sox2 overexpression. The improvement in cell proliferation and differentiation using Oct4/Sox2 expression in ATMSCs may be a useful method for expanding the population and increasing the stemness of ATMSCs.

Potential Immunoinflammatory Role of Staphylococcal Enterotoxin A in Atopic Dermatitis: Immunohistopathological Analysis and in vitro Assay

BACKGROUND: The underlying mechanism of atopic dermatitis (AD) exacerbated by Staphylococcus aureus has not been established. However, we demonstrated recently that the majority of S. aureus strains colonized in the skin of Korean AD patients carried genes encoding staphylococcal enterotoxin A (SEA) and/or toxic shock syndrome toxin-1 (TSST-1). OBJECTIVE: To clarify the role of staphylococcal superantigen, SEA in AD. METHODS: With the lesional skin of 9 AD patients and normal looking skin of one healthy adult, we examined first the expression of SEA, staphylococcal enterotoxin B (SEB), and TSST-1 using immunohistochemical analysis. In addition, we investigated the effects of SEA on the expression of inflammation-related adhesion molecules and cytokines in human HaCaT keratinocytes and Human Umbilical Vein Endothelial Cells (HUVECs) by reverse transcriptase-polymerase chain reaction (RT-PCR) analysis and enzyme-linked immunosorbent assay. RESULTS: Staphylococcal protein A (SPA) and SEA were detected with increased immunoreactivity in AD patients. However, TSST-1 showed mild-to-moderate immunoreactivity in AD patients, whereas SEB was minimally detected. In the double immunofluorescence investigation, SEA and SPA were well co-localized. SEA induced upregulation of adhesion molecules and elicited inflammatory responses in HaCaT keratinocytes and HUVECs. CONCLUSION: This study demonstrates the importance of SEA as an immunoinflammatory triggering factor of AD in Koreans.

C-reactive protein as an indicator of inflammatory responses to experimentally induced cystitis in dogs

The aim of this study was to demonstrate and assess C-reactive protein (CRP) changes in dogs with induced bacterial cystitis with or without antibiotics. We also evaluated availability of CRP levels to serve as an indicator for monitoring or diagnosing bacterial cystitis. Serial CRP concentrations in dogs with induced bacterial cystitis were higher than those of controls (p < 0.001). CRP concentrations peaked on day 7 and gradually decreased thereafter. In the treatment group, CRP concentrations decreased after medication compared to the untreated group (p = 0.032). CRP levels had a linear correlation with urine white blood cell counts among all groups (r = 0.837, p < 0.001, n = 140). Compared to the negative urine culture group, dogs with positive urine culture results had higher CRP concentrations (median 43.8 mg/L vs. 5.9 mg/L; p < 0.001). Area under the receiver operating characteristic curve was 0.955; when cut-off value was 12.2 mg/L, CRP measurements were found to have a sensitivity of 92.3% and specificity of 86.4%. This result indicates that rapid increases of CRP occurred after inducing bacterial cystitis and CRP may be a useful indicator for monitoring or diagnosing canine bacterial cystitis together with sediment urinalysis and urine bacterial culture.

Accidental Mortality and Compensation Payment in School Activities among Elementary, Middle and High School Students in Seoul over Twenty Years (1988-2007)

PURPOSE: The purpose of this study was to identify accidental mortality during school activities of students in elementary, middle or high school in Seoul and consequent compensation payment. METHODS: Fifty-eight students died due to accidents during the period 1988 through 2007. Data were obtained from the Seoul School Safety and Insurance Association, and Seoul Metropolitan Office of Education. Chi-square, t test, and ANCOVA were used in the data analysis. RESULTS: Among students, 75.9% were male and 37.9% were high school students. Accidental mortality was 1.61 per one million students (2.33 for male, and 0.82 for female students, and 0.93, 2.13 and 2.31 for elementary, middle and high school students, respectively). Mortality caused by drowning and falls per one million student was 0.85 and 0.74 for male, and 0.23 and 0.35 for female students. After age, year and cause were adjusted using ANCOVA, the mean compensation payment was 40,615 thousand won for male, and 62,000 thousands for female students. Highest compensation payment was 127,137 thousand for cerebral concussion after age, gender, year and cause were adjusted. CONCLUSION: To decrease student accidental mortality, especially drowning and falls, development of efficient safety-enforcing education is essential to prevent injuries and avoid preventable compensation costs.

A Case of Zinc Deficiency Dermatitis in Breast Milk Fed Infant Mimicking Atopic Dermatitis / 소아알레르기및호흡기학회지

Zinc deficiency can result from acrodermatitis enteropathica, an inadequate dietary intake, malabsorption, increased body loss, or intravenous feeding. Unlike acrodermatitis enteropathica, breast feeding-induced acrodermatitis is transient and improves when nursing ends. Breast feeding-induced acrodermatitis is caused by a ZnT-2 transporter dysfunction in the mother's mammary glands. We report a case of a 6-month-old male infant who presented with erythematous patches and plaques involving the perioral, perineal, and acral areas and loose stools since 2 weeks after birth. Atopic dermatitis was considered initially, but the lesions did not respond to topical antibiotics, corticosteroids, or maternal dietary restriction from allergenic food. The patient's serum zinc concentration was low, and the mother's serum and milk had low zinc levels. The differential diagnosis for atopic dermatitis is extensive. Physicians should be alert for the possibility of zinc deficiency dermatitis in breast milk fed infants that mimics atopic dermatitis.

Successful Catheter Ablation of Focal Automatic Left Ventricular Tachycardia Presented with Tachycardia-Mediated Cardiomyopathy

Non-reentrant focal tachycardias occur spontaneously, facilitated by catecholamine infusion, but they cannot be initiated or terminated with programmed stimulation. These tachycardias exhibit early activation before the QRS, however, do not typically show the mid-diastolic potential that is crucial for reentrant tachycardia maintenance. Electrophysiological studies are useful for distinguishing focal from macro-reentrant ventricular tachycardia. We report herein a case of patient without a history of structural heart disease who presented with a focal Purkinje ventricular tachycardia and heart failure. The focal Purkinje ventricular tachycardia was eliminated by radiofrequency catheter ablation. All of the patien's symptoms were improved after ablation.

Two Pediatric Osteosarcoma Cases with Delayed Methotrexate Excretion: Its Clinical Course and Management / Journal of the Korean Cancer Association, 대한암학회지

High-dose methotrexate (MTX) chemotherapy extends the duration of hospitalization and introduces the risks of serious complications related to delayed MTX excretion. The treatment of delayed MTX excretion is largely dependent on invasive measures such as hemodialysis because the clinical data regarding the efficacy or safety of carboxypetidase G2 is limited. We report here on the cases of two pediatric osteosarcoma patients with delayed MTX excretion and who were successfully managed using supportive measures. Potential life-threatening complications were prevented by administering high doses of leucovorin.

Guillain-Barre Syndrome in a Patient with Chemotherapy for HIV Related Burkitt's Lymphoma

Human immunodeficiency virus infection is not a common cause of Guillain-Barre syndrome. Guillain-Barre syndrome with cerebrospinal fluid pleocytosis has been associated with early human immunodeficiency virus (HIV) infection, occasionally as the presenting manifestation. We report a case of 73-year-old Korean malen with acute motor axonal variant of Guillain-Barre syndrome during chemotherapy for HIV- related Burkitt's lymphoma. This is the first report of Guillain-Barre syndrome occurringed within HIV infection in Korea.

Expression of IL-17A during Thymus Regeneration in the Rat / 체질인류학회지

IL-17A is a pro-inflammatroy cytokine secreted by activated T cells. The IL-17 family consist of IL-17A, IL-17B, IL-17C, IL-17D, IL-17E and IL-17F. IL-17A and IL-17F are produced primarily in activated T cells. In contrast, IL-17B, IL-17C, IL-17D and IL-17E are expressed in a wide assortment of tissues. Their functions partially overlap those of IL-17A, although they have not been as thoroughly investigated. The receptor for IL-17A (IL-17R) is widely expressed in a variety of tissues. IL-17A and IL-17E mRNAs were expressed in only EL4 cells. IL-17C mRNA expression was observed in the thymic subcapsular/cortex epithelial cells (SNEC), cortex or cortical reticular cells (CREC), medullary epithelial cells (MEC), medullary interdigitating-like cells (MDC), thymocytes and EL4 cells. However, IL-17C mRNA was not expressed in RAW 264.7 cells. Immunohistochemical study also demonstrated not only the presence of IL-17A mainly in the thymic epithelial cells, but also the upregulated expression of IL-17A in the thymic epithelial cells of the regenerating thymus. Thus, the results of the present study suggest that IL-17A expressed in the thymocytes and thymic epithelial cells could play an important role in the development of new T cells to replace T cells damaged by cyclophosphamide treatment during thymus regeneration.

Extrinsic Indentation at Gastric Fundus by Splenic Lymphangioma / 대한소화기내시경학회지

Splenic lymphangioma is a very rare benign condition, and it is classified as one of the cystic proliferations of the spleen. This is considered to result from developmental malformation of the lymphatic system. Splenic lymphangioma is usually seen in children and it is often found incidentally. Herein, we report on an unusual case of splenic lymphangioma in an adult. A 66-year-old woman presented with abdominal pain. On esophagogastroduodenoscopy, the gastric fundus was externally compressed by an extrinsic mass. Computed tomography revealed multiple cystic masses in the spleen. Laparoscopic splenectomy was then performed. The histology revealed multiple splenic lymphangiomas. This case showed an unusual presentation of splenic lymphangioma as gastric extrinsic compression, and this should be examined by imaging studies.

Late Cytomegalovirus Disease Causes Ileal Perforation after Kidney trasplantation

Cytomegalovirus (CMV) is the single most common infection following kidney transplantation and despite prophylactic strategies and the development of new antiviral agents, it still remains a cause of considerable morbidity and mortality. Current literature suggests that CMV infection may trigger rejection. We report a case of late CMV disease in a preemptive seropositive recipient who did not receive CMV prophylaxis. Diarrhea and abdominal cramping persisted after the administration of mycophenolate mofetil (MMF) six months after transplantation and resulted in ileal perforation at eight months after transplantation. The boy recovered after six weeks of treatment with ganciclovir. MMF has been mooted as a risk factor for CMV infection since its introduction, and further investigations are required to confirm its role. More attention to infectious complications is necessary and serial monitoring of viral load is recommended when MMF is administered.

The Medical Use of the Disabled Among Overusers of Medical Aid in Korea / 예방의학회지

OBJECTIVES: In Korea, the top 10% of Medical Aid recipients represent nearly 60% of total payment, with the costs for those disabled for over 365 days representing approximately 30% of total payment. The purpose of this study was to compare Medical Aid use of the disabled with non-disabled recipients, and to identify contributing factors to the total payment in the top 2% of recipients identified as Medical Aid overusers. METHODS: Subjects (n=2,211) selected were > or =18-years-of-age and received >1000 days of co-payment-free type I Medical Aid. Case managers (n=200) conducted interviews in December 2006, and collected data from Health Insurance Review & Assessment Service. Amounts over the 9 months from January September 2006 were analyzed descriptively and using Chi-square, ANCOVA, and robust multiple linear regression. RESULTS: Disabled individuals (mean age 61.3 years) composed 36.6% of subjects; 44.8% of the disabled were male. On a monthly basis per capita, the disabled group averaged 10.5 outpatient days, total payment of 523,000 Korean Won(won), inpatient payment of 359,000won, and outpatient payment of 183,000won. All values exceeded the monthly average for non-disabled individuals. Contributing factors were identified as male gender (82,000won), elementary school or lower educational level (64,000won), residence in a small city (82,000won), lack of family support (61,000won), kidney disability (673,000won), intellectual disability (151,000won), and multiple disabilities (119,000won). CONCLUSIONS: The identification of contributing factors to Medical Aid use by those defined as disabled supports the adoption of comprehensive alternative policies such as strengthening of education and consultation services, provision of alternative facilities, and promotion of self-care.

Clinical Significance of Interferon gamma Release Assay for Diagnosis of Tuberculosis in Children / 소아감염

PURPOSE: The purpose of this study is to evaluate the usefulness and limitation of QuantiFERON-TB Gold (QTB) for diagnosis of latent tuberculosis in children. METHODS: The medical records of 112 children who were tested by tuberculin skin test (TST) and QTB for detection of latent tuberculosis (TB) in Eulji General Hospital during the period from January 2007 to June 2009 were retrospectively reviewed. RESULTS: Of the 112 participants, the clinical TB group included 15 (13.4%), the contact group included 43 (38.4%), and the non-contact group included 54 (48.2%). Positive rates of TST and QTB were 100% and 80% in the clinical TB group, 64% and 14% in the close contact group, 27% and 0% in the casual contact group, and 52% and 2% in the non-contact group, respectively. Sensitivity of QTB was 80.0% and specificity was 92.6%. Agreement between QTB and TST was poor (kappa=0.209). We also confirmed that the positive rate of QTB increased as the age of the patient increased (P=0.011). A QTB indeterminate result was observed in 11 (9.8%) subjects. QTB was retested in 15 subjects. In 5 of the 6 subjects who had positive results initially, positive results persisted for a median 2.2 months after termination of treatment. CONCLUSION: Although QTB was associated with several problems, including low sensitivity and a high rate of indeterminate results, it had clinical importance due to its high specificity. We found good correlation with regard to exposure and QTB positivity, including that of young children under 5 years of age. However, clinical application of interferon-gamma releasing assay for young children for diagnosis of active and latent tuberculosis will require additional prospective studies.

The Contributing Factors to Surplus Medicine by Long-Term Users of Medical Aid in Korea / 예방의학회지

OBJECTIVES: The amount of medical utilization by Medical Aid recipients was 3.7 times that of patients with Korean Medical Insurance. This study aims to describe the surplus medicine and the medication-related utilization, and to determine factors contributing to surplus medicine. METHODS: Among those who used copayment-free Class I Medical Aid in 2005, 146,880 subjects who were > or =19 year-old and received >365 days medical treatment per year were studied with their case managers by conducting face-to-face interviews. The analytic methods were description, chi-square, t-tests, ANCOVA and multiple logistic regressions. RESULTS: Most subjects were female (68.6%), the elderly (62.5%), and the separated (61.6%), had an elementary graduation or less (74.8%), and had disabilities (33.2%). The percentage of subjects with surplus medicine was 18.5%. However, the percentage of females, the elderly, those with non-disabilities, the separated, the uneducated, those with a very poor perceived health status and those with an economical burden for medical treatment was 19.3%, 18.9%, 19.0%, 19.3%, 19.0%, 20.2% and 24.3%, respectively. For subjects with surplus medicine, averages for the number of used pharmacies, the pharmacy-visit days and the medication costs were 4.6 drugstores, 34.9 days and approximately 1,124 thousand Won. These values were higher than those without surplus medicine (4.4 drugstores , 33.8 days, and 1,110 thousand won, respectively). The odds ratios of the contributing factors to surplus medicine were female 1.11 (95% CI=1.07-1.14), the elderly 1.06 (95% CI=1.02-1.10), those with non-disabilities 1.08 (95% CI=1.05-1.12), the separated 1.14 (95% CI=1.10-1.18), the unmarried 1.12 (95% CI=1.07-1.18), the uneducated 1.03 (95% CI=1.01-1.08), those with a very poor perceived health status 1.04 (95% CI=1.01-1.08) and experiencing an economical burden for medical treatment 2.33 (95% CI=2.26-2.40). CONCLUSIONS: 18.5% of subjects had surplus medicine with a higher mean of medication cost. Therefore, health education and health promotion programs to prevent surplus medicine and to improve the appropriate usage of medication are necessary.

Expression of 4-1BB and 4-1BBL in thymocytes during thymus regeneration

4-1BB, a member of the tumor necrosis factor receptor (TNFR) superfamily, is a major costimulatory receptor that is rapidly expressed on the surface of CD4+ and CD8+ T cells after antigen- or mitogen-induced activation. The interaction of 4-1BB with 4-1BBL regulates immunity and promotes the survival and expansion of activated T cells. In this study, the expression of 4-1BB and 4-1BBL was examined during regeneration of the murine thymus following acute cyclophosphamide-induced involution. Four-color flow cytometry showed that 4-1BB and 4-1BBL were present in the normal thymus and were preferentially expressed in the regenerating thymus, mainly in CD4+CD8+ double-positive (DP) thymocytes. Furthermore, the CD4loCD8lo, CD4+CD8lo and CD4loCD8+ thymocyte subsets, representing stages of thymocyte differentiation intermediate between DP and single-positive (SP) thymocytes, also expressed 4-1BB and 4-1BBL during thymus regeneration but to a lesser degree. Interestingly, the 4-1BB and 4-1BBL positive cells among the CD4+CD8+ DP thymocytes present during thymus regeneration were TCR(hi) and CD69+ unlike the corresponding controls. Moreover, the 4-1BB and 4-1BBL positive cells among the intermediate subsets present during thymus regeneration also exhibited TCRhi/int and CD69+/int phenotypes, indicating that 4-1BB and 4-1BBL are predominantly expressed by the positively selected population of the CD4+CD8+ DP and the intermediate thymocytes during thymus regeneration. RT-PCR and Western blot analyses confirmed the presence and elevated levels of 4-1BB and 4-1BBL mRNA and protein in thymocytes during thymus regeneration. We also found that the interaction of 4-1BB with 4-1BBL promoted thymocyte adhesion to thymic epithelial cells. Our results suggest that 4-1BB and 4-1BBL participate in T lymphopoiesis associated with positive selection during recovery from acute thymic involution.

Prevalence of Bartonella henselae and Bartonella clarridgeiae in cats and dogs in Korea

Blood, saliva, and nail samples were collected from 54 dogs and 151 cats and analyzed for the presence of Bartonella henselae with a novel nested polymerase chain reaction (PCR) method. Bartonella (B.) henselae was detected in feral cat blood (41.8%), saliva (44.1%), and nail (42.7%) samples. B. henselae was also detected in pet cat blood (33.3%), saliva (43.5%), and nail (29.5%) samples and in pet dog blood (16.6%), saliva (18.5%), and nail (29.6%) samples. Nine samples were infected with B. clarridgeiae and 2 were co-infected with B. henselae and B. clarridgeiae of blood samples of dogs. This report is the first to investigate the prevalence of B. henselae and B. clarridgeiae in dogs and cats in Korea, and suggests that dogs and cats may serve as potential Bartonella reservoirs.