In-situ Hybridization for the Detection of Sacbrood Virus in Infected Larvae of the Honey Bee (Apis cerana).

The aim of this study was to develop and use in-situ hybridization (ISH) for the detection and localization of the sacbrood virus (SBV) in Korean honey bee (Apis cerana) larvae that were infected naturally with SBV. A 258 base pair cDNA probe for SBV was generated by polymerase chain reaction. Cells positive for viral genome typically showed a dark brown reaction in the cytoplasm. SBV was detected consistently in trophocytes and urocytes. The ISH was successfully applied to routinely fixed and processed tissues and thus should prove helpful in the diagnosis and characterization of viral distribution in infected larvae.

Rectus abdominis muscle resection and fascial reconstruction for the treatment of uterine leiomyosarcoma invading the abdominal wall: a case report.

The authors present a case of intra-abdominal recurrent leiomyosarcoma invading a large area of the abdominal wall. The patient underwent cytoreductive surgery, including resection of the rectus abdominis muscle, followed by reconstruction of the defect using synthetic mesh. The tumor was surgically removed by en bloc resection, including most of the rectus abdominis muscle and ileum. The abdominal wall defect was repaired using synthetic mesh. The patient underwent radiotherapy and chemotherapy after the surgery and was healthy one year later.

Cost minimization analysis to compare activated prothrombin complex concentrate (APCC) and recombinant factor VIIa for haemophilia patients with inhibitors undergoing major orthopaedic surgeries.

Benefits of bypassing agents for maintaining haemostasis in major surgeries have been described in the literature; however, their use has a substantial economic impact. This study assessed the cost of FEIBA, an activated prothrombin complex concentrate and recombinant factor VIIa (rFVIIa) when used in inhibitor patients undergoing major surgeries. After reviewing published literature, a cost minimization model was developed describing dosing regimens recommended and used during major surgeries for FEIBA (pre-operative: 75-100 U kg(-1); postoperative: 75-100 U kg(-1) q 8-12 h days 1-5 and 75-100 U kg(-1) q 12 h days 6-14) and rFVIIa (pre-operative: 90 microg kg(-1); intra-operative: 90 microg kg(-1) q 2 h; postoperative: 90 microg kg(-1) q 2-4 h days 1-5 and 90 microg kg(-1) q 6 h days 6-14). Using a 75 kg patient and US prices, total drug cost was calculated for three scenarios: use of FEIBA or rFVIIa alone and a third case combining rFVIIa pre- and intra-operative and FEIBA throughout a 14-day postoperative period. Dosage amounts of modelled bypassing agents were similar to cases in the literature. Using FEIBA instead of rFVIIa would decrease total drug cost by >50% and save over $400,000 per surgery. Sequential use of both bypassing agents would increase total drug cost by 9% when compared with FEIBA alone, but would remain >40% lower than rFVIIa alone. Univariate sensitivity analyses confirmed robustness of results. As large amounts of bypassing agents are necessary for patients with inhibitors to undergo major surgeries, cost is a major consideration. Use of FEIBA alone or in combination with rFVIIa has emerged as a cost-saving approach.

Expression of human leukocyte antigen-G and its correlation with interleukin-10 expression in cervical carcinoma.

OBJECTIVE: To determine human leukocyte antigen (HLA)-G and interleukin (IL)-10 expression and the relationship between HLA-G expression and clinicopathologic features in patients with cervical cancer. METHOD: Tissue samples were obtained from 40 patients with cervical cancer and 15 control patients with a normal cervix. Quantitative real-time RT-PCR for mRNA and western blot analysis for protein expression were used. RESULT: Both HLA-G and IL-10 mRNA expression in cervical cancer tissues was significantly greater than normal controls (P<0.001, P=0.005). Protein expression of HLA-G and IL-10 in the cancer group was also significantly greater than in the controls (P<0.001, P=0.021). There was an inverse relationship between FIGO stage and HLA-G mRNA expression (P=0.046). CONCLUSION: HLA-G and IL-10 might play an important role in cancer progression of the cervix. High HLA-G mRNA expression may be related to early carcinogenesis since it was associated with early-stage cervical cancer.

Prognostic factors and expression of p53 and mdm-2 in uterine sarcomas.

OBJECTIVES: To evaluate the clinicopathologic prognostic factors of uterine sarcoma, and determine whether overexpression of p53 and mdm-2 proteins correlate with overall survival and prognostic factors. METHODS: Thirty specimens from 43 patients with uterine sarcoma were available for slide review and evaluated by immunohistochemistry at Yonsei and Ajou University Medical Centers for the expression of p53 and mdm-2. RESULTS: Overall survival was found to correlate to histologic type of uterine sarcoma, surgical stage, tumor grade, and mitotic index. Whereas 63% of the sarcomas expressed p53, with a significant correlation to overall survival, 60% expressed mdm-2, with a significant correlation to the mitotic index but not to overall survival. CONCLUSION: Histologic type, surgical stage, tumor grade, mitotic index, and p53 expression were prognostic factors of the overall survival of patients with uterine sarcoma.

Expression profiling of cyclin B1 and D1 in cervical carcinoma.

AIM: Cyclins are a family of regulatory proteins that play a key role in controlling the cell cycle. Abnormalities of cell cycle regulators, including cyclins and cyclin dependent kinases, have been reported in various malignant tumors. This study was undertaken to quantitatively detect cyclin B1 and D1 in cervical cancer. METHODS: A quantitative real-time reverse transcription polymerase chain reaction and Western blot assay were used to analyze the expression of cyclin B1/D1 mRNA and proteins, respectively, in fresh invasive cervical cancer (n = 41) and normal cervical tissues (n = 10). RESULTS: There was significantly greater cyclin B1 expression in invasive cervical cancer than in normal cervical tissue (P = 0.019). However, cyclin D1 expression was not significantly different. A Western blot assay yielded similar results. CONCLUSION: Our results were consistent with the concept that up-regulation of cyclin B1 expression occurred in cervical cancer and an aberrant expression of cyclin B1 might play an important role in cervical carcinogenesis.

Pretreatment levels of serum squamous cell carcinoma antigen and urine polyamines in women with squamous cell carcinoma of the cervix.

OBJECTIVES: To investigate whether pretreatment levels of serum squamous cell carcinoma antigen (SCCA) and urine polyamines can predict lymph node metastases in patients with early stage cervical carcinoma. METHODS: Pretreatment measurement of serum SCCA and urine polyamine levels was carried out for 419 women. Of those women, 104 with stage IB and IIA cancer received radical surgery and had tumor size reassessed postoperatively. RESULTS: The women had increased levels of serum SCCA (>2.0 ng/mL) and elevated urine polyamines (>45 micromol/g of creatinine) with advanced cancer stage (P<0.01). The median SCCA level was significantly higher in women with metastatic disease than that in those without lymph node involvement (3.9 vs. 1.1; P<0.01). Women with nodal involvement also had significantly higher median levels of urine polyamines than those without nodal disease. CONCLUSIONS: Pretreatment measurement of SCCA and urine polyamine levels may help in predicting lymph node metastases in women with early stage cervical carcinoma.

The type I BMP receptor BmprIB is essential for female reproductive function.

Maintenance of female reproductive competence depends on the actions of several hormones and signaling factors. Recent reports suggest roles for bone morphogenetic proteins (BMPs) in early stages of folliculogenesis. A role for the type I BMP receptor BmprIB as a regulator of ovulation rates in sheep has been described recently, but little is known about the roles of BMP signaling pathways in other aspects of reproductive function. We report here that BMPRIB is essential for multiple aspects of female fertility. Mice deficient in BmprIB exhibit irregular estrous cycles and an impaired pseudopregnancy response. BmprIB mutants produce oocytes that can be fertilized in vitro, but defects in cumulus expansion prevent fertilization in vivo. This defect is associated with decreased levels of aromatase production in granulosa cells. Unexpectedly, levels of mRNA for cyclooxygenase 2, an enzyme required for cumulus expansion, are increased. BmprIB mutants also exhibit a failure in endometrial gland formation. The expression of BmprIB in uterine linings suggests that these defects are a direct consequence of loss of BMP signaling in this tissue. In summary, these studies demonstrate the importance of BMP signaling pathways for estrus cyclicity, estradiol biosynthesis, and cumulus cell expansion in vivo and reveal sites of action for BMP signaling pathways in reproductive tissues.

Specific binding of recombinant Listeria monocytogenes p60 protein to Caco-2 cells.

The Listeria monocytogenes p60 is a major extracellular protein, which is believed to be involved in the invasion of these bacteria into their host cells. So far the mechanism by which p60 participates in the internalization or penetration of L. monocytogenes is still veiled. To determine the possibility of a direct interaction of p60 with the host cell surface, the iap gene was recombinantly expressed in Escherichia coli and used for binding studies with the enterocyte-like Caco-2 cells. Fluorescence activated flow cytometry and confocal laser scanning microscopy revealed a cell membrane specific staining with p60, which implications in Listeria virulence are discussed.