Synergistic Anti-bacterial Effects of Phellinus baumii Ethyl Acetate Extracts and beta-Lactam Antimicrobial Agents Against Methicillin-Resistant Staphylococcus aureus

BACKGROUND: The development of new drugs or alternative therapies effective against methicillin-resistant Staphylococcus aureus (MRSA) is of great importance, and various natural anti-MRSA products are good candidates for combination therapies. We evaluated the antibacterial activities of a Phellinus baumii ethyl acetate extract (PBEAE) and its synergistic effects with beta-lactams against MRSA. METHODS: The broth microdilution method was used to determine the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of the PBEAE. The PBEAE synergistic effects were determined by evaluating the MICs of anti-staphylococcal antibiotic mixtures, with or without PBEAE. Anti-MRSA synergistic bactericidal effects of the PBEAE and beta-lactams were assessed by time-killing assay. An ELISA was used to determine the effect of the PBEAE on penicillin binding protein (PBP)2a production. RESULTS: The MICs and MBCs of PBEAE against MRSA were 256-512 and 1,024-2,048 microg/mL, respectively. The PBEAE significantly reduced MICs of all beta-lactams tested, including oxacillin, cefazolin, cefepime, and penicillin. However, the PBEAE had little or no effect on the activity of non-beta-lactams. Time-killing assays showed that the synergistic effects of two beta-lactams (oxacillin and cefazolin) with the PBEAE were bactericidal in nature (Deltalog10 colony forming unit/mL at 24 hr: 2.34-2.87 and 2.10-3.04, respectively). The PBEAE induced a dose-dependent decrease in PBP2a production by MRSA, suggesting that the inhibition of PBP2a production was a major synergistic mechanism between the beta-lactams and the PBEAE. CONCLUSIONS: PBEAE can enhance the efficacy of beta-lactams for combined therapy in patients infected with MRSA.

Characterization of Two Novel mAbs Recognizing Different Epitopes on CD43

JL1, a specific epitope on CD43, is a potential biomarker for the diagnosis of acute leukemia. Although qualitative assays for detecting leukemia-specific CD43 exist, there is a need to develop quantitative assays for the same. Here, we developed two novel monoclonal antibodies (mAbs), 2C8 and 8E10, recognizing different epitopes on CD43. These clones are capable of pairing with YG5, another mAb against JL1 epitope, because they were selectively obtained using sandwich ELISA. Antigens recognized by 2C8 and 8E10 were confirmed as CD43 by western blotting using the CD43-hFC recombinant protein. When expression on various leukemic cell lines was investigated, 2C8 and 8E10 displayed a disparity in the distribution of the epitope. Enzyme assays revealed that these mAbs recognized a sialic acid-dependent epitope on CD43. Using normal thymus and lymph node paraffin-embedded tissues, we confirmed a difference in the epitopes recognized by the two mAbs that was predicted based on the maturity of the cells in the tissue. In summary, we developed and characterized two mAbs, 2C8 and 8E10, which can be used with YG5 in a sandwich ELISA for detecting leukemia-specific CD43.

Evaluation of Urine Cytology in Urothelial Carcinoma Patients: A Comparison of CellprepPlus(R) Liquid-Based Cytology and Conventional Smear

BACKGROUND: Urine cytology is an important test in the screening of urothlelial neoplasms. The conventional smear (CS) method of testing urine samples has a low sensitivity, approximately 50% result accuracy for detecting urothelial carcinomas, while liquid-based cytology (LBC) has much improved diagnostic accuracy, sensitivity, and specificity. The aim of this study was to compare the morphologic features and diagnostic efficacy of CellprepPlus(R) LBC with those of CS for urine cytology. METHODS: A total of 713 cases of urine specimens collected from November 2009 to September 2010 were included. All specimens were divided equally for the preparation of CellprepPlus(R) LBC and CS for each case. RESULTS: CellprepPlus(R) revealed more cellularity, a cleaner background and better cytomorphologic features, but it showed a less intact architectural pattern compared to that of CS. Of the 88 histologically confirmed cases, the diagnostic sensitivity for CellprepPlus(R) was 50% and higher than the 37.5% for CS. The specificity of both preparations was 100%. CONCLUSIONS: The CellprepPlus(R) showed an improved quality of slides and provided better diagnostic accuracy, thus CellprepPlus(R) could be a first-line screening tool in urinary tract cytology.

Generation of 1E8 Single Chain Fv-Fc Construct Against Human CD59

BACKGROUND: Therapeutic approaches using monoclonal antibodies (mAbs) against complement regulatory proteins (CRPs:i.e.,CD46,CD55 and CD59) have been reported for adjuvant cancer therapy. In this study, we generated a recombinant 1E8 single-chain anti-CD59 antibody (scFv-Fc) and tested anti-cancer effect.by using complement dependent cytotoxicity (CDC). METHODS: We isolated mRNA from 1E8 hybridoma cells and amplified the variable regions of the heavy chain (VH) and light chain (VL) genes using reverse-transcriptase polymerase chain reaction (RT-PCR). Using a linker, the amplified sequences for the heavy and light chains were each connected to the sequence for a single polypeptide chain that was designed to be expressed. The VL and VH fragments were cloned into the pOptiVEC-TOPO vector that contained the human CH2-CH3 fragment. Then, 293T cells were transfected with the 1E8 single-chain Fv-Fc (scFv-Fc) constructs. CD59 expression was evaluated in the prostate cancer cell lines using flow cytometry. The enhancement of CDC effect by mouse 1E8 and 1E8 scFv-Fc were evaluated using a cytotoxicity assay. RESULTS: The scFv-Fc constructs were expressed by the transfected 293T cells and secreted into the culture medium. The immunoreactivity of the secreted scFv-Fc construct was similar to that of the mouse 1E8 for CCRF-CEM cells. The molecular masses of 1E8 scFv-Fc were about 120 kDa and 55 kDa under reducing and non-reducing conditions, respectively. The DNA sequence of 1E8 scFv-Fc was obtained and presented. CD59 was highly expressed by the prostate cancer cell line. The recombinant 1E8 scFv-Fc mAb revealed significantly enhanced CDC effect similar with mouse 1E8 for prostate cancer cells. CONCLUSION: A 1E8 scFv-Fc construct for adjuvant cancer therapy was developed.

Seminal Vesicle Involvement by Urothelial Carcinoma in Situ of the Bladder with Mucosal Spread Pattern: A Case Report

Mucosal spreading of urothelial tumors to the seminal vesicles is very rare. We experienced a case of mucosal involvement of the seminal vesicles by a bladder tumor in a 72-year-old man. The patient had a history of transurethral resection for invasive urothelial carcinoma of the bladder 8 years previously. Radical cystoprostatectomy was performed owing to recurrent and multiple urothelial carcinoma in situ. Microscopically, the urothelial carcinoma in situ was throughout the mucosa of the urinary bladder, both ureters, the prostate, and the left seminal vesicle. To date, the implication of mucosal involvement of the seminal vesicles by urothelial carcinoma is unclear. However, careful microscopic examination is needed to avoid an erroneous diagnosis.

Polypoid cystitis in an adult without history of catheterization

Polypoid cystitis is a benign exophytic mucosal lesion of the bladder. Differentiating it from papillary transitional cell carcinoma is difficult due to their similar characteristics. Although indwelling catheter is the main well-known cause of polypoid cystitis, some case reports unrelated to catheterization have been described. However, the radiological findings of polypoid cystitis have rarely been reported. We hereby describe polypoid cystitis in a 20-year-old man without a history of catheterization along with the computed tomographic findings

Characterization of Monoclonal Antibodies against Human Leukocyte Common Antigen (CD45)

BACKGROUND: The leukocyte common antigen (CD45) is a transmembrane-type protein tyrosine phosphatase that has five isoforms. METHODS: We generated seven murine mAbs against human CD45 by injecting cells from different origins, such as human thymocytes, PBMCs, and leukemic cell lines. By using various immunological methods including flow cytometry, immunohistochemistry, and immunoprecipitation, we evaluated the reactivity of those mAbs to CD45 of thymus as well as tonsil lysates. Furthermore, we transiently transfected COS-7 cells with each of gene constructs that express five human CD45 isoforms respectively, and examined the specificities of the mAbs against the transfected isoforms. RESULTS: In case of thymocytes, lymphocytes, and monocytes, all the seven mAbs demonstrated positive reactivities whereas none was reactive to erythrocytes and platelets. The majority of immune cells in formalin-fixed paraffin-embedded thymus and tonsil tissues displayed strong membranous immunoreactivity, and the main antigen was detected near 220 kDa in all cases. Among the mAbs, four mAbs (AP4, DN11, SHL-1, and P6) recognized a region commonly present in all the five isoforms. One mAb, YG27, recognized four isoforms (ABC, AB, BC, and O). Two mAbs, P1 and P14, recognized the isoforms that contain exon A encoded regions (ABC and AB). CONCLUSION: In this study, we confirmed that AP4, DN11, SHL-1, YG27 and P6, are mAbs reactive with the CD45 antigen whereas P1 and P14 are reactive with the CD45RA antigen.

Squamous Cell Carcinoma and Basal Cell Carcinoma Simultaneously Arising in a Nevus Sebaceus: A Case Report

PURPOSE: Nevus sebaceus is a congenital hamartomatous lesion, typically involving head and neck. Various benign and malignant neoplasms can develop in association with nevus sebaceus. We report a case of simultaneous occurrence of squamous cell carcinoma and basal cell carcinoma in nevus sebaceus. METHODS: A 73-year-old man presented with erythematous to black verrucous nodules on the right subauricular area. The upper part was accompanied with inflammation and ulceration, and no specific findings suspicious for malignant degeneration were found in the lower part preoperatively. The mass was totally excised and the defect was directly closed. Lymph nodes were not involved on concomitant neck dissection. RESULTS: Histopathologic examination confirmed the presence of squamous cell carcinoma in the upper part and basal cell carcinoma in the lower part of a nevus sebaceus. Negative margins were achieved on resection. No clinical problems were found during the 3-month follow-up period. CONCLUSION: We experienced a rare case of simultaneous occurrence of squamous cell carcinoma and basal cell carcinoma within the same nevus sebaceus. Because patients with nevus sebaceus have risk of malignant changes, surgical excision and work-up for recurrence and metastasis should be considered in suspicious cases. And even in a totally asymptomatic case, the possibility of occult secondary cancer should be informed before surgery in aged patients.

Characterization of a Novel Monoclonal Antibody (27H2) Recognizing Human CD34 Class III Epitope

BACKGROUND: Monoclonal antibodies (mAbs) recognizing Class III epitope of CD34 are essential for flow cytometric diagnosis of leukemia. METHODS: 27H2 mAb was developed from a mouse alternatively immunized with human acute leukemia cell lines, KG1 and Molm-1. Using flow cytometric analysis of various leukemic cell lines and peripheral blood, immunohistochemical study of frozen tonsil, we characterized 27H2 mAb. Antigen immunoprecipitated with 27H2 mAb immunobloted with anti-CD34 mAb. A case of bone marrow sample of acute lymphoblastic leukemia (ALL) patient was obtained at CBNU Hospital. For epitope identification enzyme treatment with neuraminidase and O-sialoglycoprotein endopeptidase (OSGE) and blocking assay with known classIII mAb (HPCA-2) were done. RESULTS: Only KG1 and Molm-1 revealed positive immunoreactivity. Immunohistochemical staining disclosed strong membranous immunoreactivity on high endothelial venules. Antigen immunoprecipitated by 27H2 mAb showed approximately 100 kDa sized band immunoblotted with anti-CD34 under non-reducing conditions. Epitope recognized by 27H2 mAb disclosed resistancy to both neuraminidase and OSGE treatment and completely blocked with known class III mAb preincubation. CD34 positive leukemic cells in BM of pre B cell ALL patient detected by FITC-conjugated 27H2 and HPCA-2 were identified with similar sensitivity. CONCLUSION: A novel murine mAb recognizing class III epitope of human CD34 with high affinity, which is useful for flow cytometric diagnosis of leukemia, was developed.

Comparison of Cytologic Evaluation between Conventional Method and CellprepPlus(R) Liquid-Based Cytology in Body Fluid

BACKGROUND: Assessment of body fluid cytology is a useful means of evaluating a metastatic tumor. Liquid-based cytology (LBC) has been developed as a replacement for the conventional Papanicolaou (CP) test. This study was performed to compare CellprepPlus(R) LBC with CP in cytologic diagnosis. METHODS: Body fluid samples (n=188, including 72 peritoneal fluid and 116 pleural fluid samples) were divided equally and analyzed by both CellprepPlus(R) and CP. RESULTS: CellprepPlus(R) revealed distributed thin layers of non-overlapping cells. All CellprepPlus(R) preparations were adequate, while 18 (9.57%) CP preparations were inadequate. The respective diagnostic rates of CellprepPlus(R) and CP were 75.0% and 76.1% negative, 10.6% and 6.38% atypical, 5.85% and 2.66% suspicious, and 8.51% and 5.32% malignant. Of the 58 confirmed cases, the sensitivity of CellprepPlus(R) and CP was 94.4% and 73.3%, respectively, and the negative predictive value was 97.2% and 87.9%, respectively. CONCLUSIONS: CellprepPlus(R) LBC has better sensitivity and negative predictive value, and produces higher quality slide preparations than than CP, making it suitable as in screening of body fluid as a cytologic diagnostic tool.

Myoepithelial Carcinoma Originated from the Maxillary Sinus / 대한이비인후과학회지

Myoepithelial carcinoma is a very rare malignant tumor accounting for less than one percent of the salivary gland neoplasms. This uncommom tumor, showing almost exclusively tumor cells with myoepithelial differentiation, is most commonly diagnosed in the parotid gland and in the minor silivary glands of the palate. We report a 77-year-old woman with myoepithelial carcinoma originating from maxillary sinus, one of the most unusual locations. The patient presented with progressive nasal obstruction and signs of a space-occupying lesion in the left maxillary sinus. A frozen biopsy identified a malignant tumor and a radical maxillectomy was performed. Histology confirmed the presence of a malignant myoepithelial carcinoma. Patient was followed up for one year without any recurrence or distant metastasis.

Cytologic Features of Epithelial-Myoepithelial Carcinoma Occurring in External Auditory Canal: A Case Report / 대한세포병리학회지

Epithelial-myoepithelial carcinoma (EMC) is a low grade malignant neoplasm that commonly occurs in the parotid gland. Recently, we investigated a case of EMC that occurred in the external auditory canal (EAC) in a 35-year-old male, and this tumor was initially diagnosed as pleomorphic adenoma. The difficulty associated with diagnosing EMC by aspiration cytology arises from both the rare incidence of this tumor and the overlapping spectrum of cytological features found in various salivary gland tumors, such as pleomorphic adenoma, basal cell adenoma and adenoid cystic carcinoma.

Cytomegalovirus Ileitis in Early Period after Kidney Transplantation

One of the most frequent viral infections after kidney transplantation (KT) is cytomegalovirus (CMV) infection. CMV infection causes disease in lung, liver, retina, brain and GI tract and reduce graft survival rate after KT. CMV disease in GI tract is most common in the colon and ileum. CMV enteritis can present with vague abdominal pain, hemorrhage and overt bowel perforation. CMV enteritis in the upper GI tract and colon can be detected early by endoscopy, but it is difficult to diagnose CMV enteritis in the small bowel. We report a rare case with CMV ileitis after KT, who gave up a transplanted kidney at post-operative 3 months in spite of the treatment of CMV infection.

A Case of Jejunal Infarction and Perforation due to Acute Pancreatitis / 대한소화기학회지

Jejunal infarction as a complication of acute pancreatitis is not common and can not be well recognized. This jejunal infarction usually arises from the venous thrombosis rather than arterial thrombosis. Jejunal infarction results in bowel perforation or stenosis according to its extension of injury and progression rate. Pathologic findings of the involved jejunum show a segmental transmural infarction and mesenteric venous thrombotic occlusions. Early diagnosis should be made for better prognosis. We report a patient with jejunal infarction resulting perforation due to acute pancreatitis, in which the initial presenting symptoms were hematemesis and abdominal distention.

Myeloid Sarcoma of Both Kidneys, the Brain, and Multiple Bones in a Nonleukemic Child

A myeloid sarcoma (MS) is an extramedullary tumor consisting of primitive granulocytic precursor cells. Although most such tumors have been reported in patients with acute myelogenous leukemia, MS is rarely recognized as an isolated tumor without any evidence of leukemia. However, in such cases, the initial diagnosis of MS can be difficult, so initial misdiagnosis rates of up to 75% have been reported. This report describes an unusual case of MS in a 3-year 5-month-old girl presenting as bilateral renal enlargements, and brain masses, with multiple bone involvements, but no hematological abnormalities.

Metastasis-suppressor KAI1/CD82 induces homotypic aggregation of human prostate cancer cells through Src-dependent pathway

To investigate the functional role of KAI1/CD82, a metastasis suppressor for human prostate cancer, in the regulation of homotypic cell adhesion, we transfected KAI1 cDNA into DU 145 human prostate cancer cells and established stable transfectant clones with high KAI1/CD82 expression. The KAI1 transfectant cells exhibited significantly increased homotypic cell aggregation in comparison with the control transfectant cells. This aggregation of the KAI1 transfectants was further enhanced upon exposure to anti-CD82 antibody, suggesting that KAI1/CD82 may be involved in the intracellular signaling for the cell adhesion. Among several signal pathway inhibitors tested, PP1, an inhibitor of Src family kinases, significantly suppressed homotypic aggregation of the KAI1 transfectant cells. Ligation of KAI1/CD82 with anti-CD82 antibody increased endogenous Src kinase activity of the KAI1 transfectant cells. When different types of src expression constructs were retransfected into the KAI1-transfected DU 145 cells, kinase-negative mutant src transfectant cells exhibited much lower homotypic aggregation than the mock cells transfected with an empty vector. Moreover, homotypic aggregation of the mutant src transfectant cells was not enhanced by KAI1/CD82 ligation with anti- CD82 antibody. These results suggest that Src mediates the intracellular signaling pathway of KAI1/CD82 for the induction of homotypic adhesion of human prostate cancer cells.

Extramedullary Plasmacytoma of the Gallbladder: A Case Report

Extramedullary plasmacytoma of the gallbladder is extremely rare. A review of the literature in English revealed only one case, involving a patient with initial extramedullary plasmacytoma of the maxillary sinus. In this report, we describe the case of a 63-year-old man with surgically confirmed extramedullary plasmacytoma confined to the gallbladder. At ultrasonography and computed tomography, diffuse and uniform thickening of the gallbladder wall without evidence of surrounding tissue infiltration was depicted.

Altered Integrin Expression Related to Anaplastic Transformation of Human Myeloma-Derived Cell Line / 대한면역학회지

No abstract available.

A study of gingival blood flow using laser Doppler flowmetry / 대한치주과학회지

The periodontal health has been evaluated clinically by various epidemiological indices, and in researches by measurement of gingival crevicular fluid. Laser Doppler flowmetry is a reliable and objective method that allows immediate measurement of erythrocyte flux in approximately one cubic mm of the capillary bed without disturbing the tissues. The purpose of the present study was to determine whether human gingival blood flow was different according to measuring area, measuring time, and sex or not. Forty volunteers with good general and periodontal health, aged early twenties and unmarried, were selected. Laser Doppler flowmetry(floLAB(R), Moor Instruments Ltd., England) was applied to measure the gingival blood flow of marginal gingiva, interdental papilla, attached gingiva and alveolar mucosa. The blood flow of interdental papilla was measured at 9-10 AM, 1-2 PM, and 5-6 PM. The difference of blood flow according to measuring area and measuring time was statistically analyzed by one way ANOVA and Dunkan test, and the difference of blood flow between men and women was statistically analyzed by t-test. (1) Mean blood flow was significantly higher in alveolar mucosa than in the gingiva(p0.1). (2) Mean blood flow was significantly higher at 5-6 PM than at 9-10 AM and 1-2 PM(p0.1). (3) There was no significant difference in gingival blood flow between men and women(p>0.1). The above results suggest that the measurment of gingival blood flow using laser Doppler flowmetry may be clinically applicable to early determination of gingival inflammation and evaluation of healing status, but further studies are necessary to standardize and simplify the measuring procedure.

Proliferative Activity of Thyroid Lesions Evaluated by Mitotic Count and Proliferating Cell Nuclear Antigen (PCNA)

To evaluate the clinical and histopathological significance of the proliferative activity in neoplastic and non-neoplastic thyroid lesions, we analyzed the mitotic count and the proliferating cell nuclear antigen labeling index (PCNA-LI) by immunohistochemistry as the proliferation- related markers. In this study included were surgically removed normal thyroid tissue (27 cases), adenomatous goiter (15 cases), Hashimoto's thyroiditis (5 cases), follicular adenoma (13 cases), follicular carcinoma (7 cases), papillary carcinoma (44 cases), poorly differentiated carcinoma (2 cases) and undifferentiated carcinoma (3 cases). The median PCNA-LI was 0 in normal thyroid tissue, 0.5 in adenomatous goiter, 6.2 in Hashimoto's thyroiditis, 1.2 in follicular adenoma, 4.8 in follicular carcinoma, 8.5 in papillary carcinoma, 60.8 in poorly differentiated carcinoma, and 55.2 in undifferentiated carcinoma (p=0.0001). Although PCNA-LI was exceptionally high in Hashimoto's thyroiditis, it was suggested that PCNA-LI could be used as a marker differentiating benign lesions from malignant neoplasm. Also, it could differentiate follicular adenoma from follicular carcinoma. Except clinical stage (p=0.0397), PCNA-LI was not related with sex, size, histologic subtype, and lymph node metastasis in papillary carcinoma. The presence of mitosis differentiated the neoplastic thyroid lesions from the non-neoplastic lesions (p<0.05), however, it could not divide benign and malignant neoplasm. These results suggest that an evaluation of the proliferative activity can help to differentiate the thyroid lesions. In addition, there was no significant correlation between the value of PCNA-LI and the presence of mitosis. It can be recommended to evaluate both the mitotic count and the PCNA-LI for determining the proliferative activity of the thyroid lesions.