Global microRNA profiles in cervical squamous cell carcinoma depend on Drosha expression levels.

Gain of chromosome 5p is seen in over 50% of advanced cervical squamous cell carcinomas (SCCs), although the genes responsible for the selective advantage provided by this abnormality are poorly understood. In the W12 cervical carcinogenesis model, we observed that 5p gain was rapidly selected over approximately 15 population doublings and was associated with the acquisition of a growth advantage and invasiveness. The most significantly upregulated transcript following 5p gain was the microRNA (miRNA) processor Drosha. In clinically progressed cervical SCC, Drosha copy-number gain was seen in 21/36 clinical samples and 8/10 cell lines and there was a significant association between Drosha transcript levels and copy-number gain. Other genes in the miRNA processing pathway, DGCR8, XPO5 and Dicer, showed infrequent copy-number gain and over-expression. Drosha copy-number and expression were not elevated in pre-malignant cervical squamous intraepithelial lesions. Importantly, global miRNA profiling showed that Drosha over-expression in cervical SCC appears to be of functional significance. Unsupervised principal component analysis of a mixed panel of cervical SCC cell lines and clinical specimens showed clear separation according to Drosha over-expression. miRNAs most significantly associated with Drosha over-expression are implicated in carcinogenesis in other tissues, suggesting that they regulate fundamental processes in neoplastic progression. Our evidence suggests that copy-number driven over-expression of Drosha and consequent changes in miRNAs are likely to be important contributors to the selective advantage provided by 5p gain in cervical neoplastic progression.

Continuing medical education-driven skills acquisition and impact on improved patient outcomes in family practice setting.

BACKGROUND: An abundance of educational theory, design, and delivery of continuing medical education (CME) learning interventions, including their impact on learners, are described in the health and social sciences literature. However, establishing a direct correlation between the acquisition of new skills by learners and patient outcomes as a result of a planned CME learning intervention has been difficult to demonstrate. METHODS: The learning intervention described here tested the impact of an injection skills-acquisition program for family physicians treating osteoarthritis of the knee by measuring patient outcomes using the pain and function subscales of the Western Ontario and McMaster (WOMAC) 3.0 osteoarthritis index, a standardized and fully validated patient-centered outcome measurement. It was hypothesized that patients of family physicians who participated in this skills-acquisition CME program would benefit from treatment administered by their physician during the time between injection skills acquisition to 6 weeks post-injection. Inclusion of a validated health status measure administered pre- and post-injection in addition to more traditional faculty and participant program evaluations was deemed necessary to test this hypothesis. Rheumatology, orthopedic surgery, and family medicine specialists from across Canada were invited to contribute to the planning, curriculum elaboration, and delivery of the viscosupplement injector preceptorship (VIP) program. Thirty-nine orthopedic and rheumatology specialists agreed to serve as expert faculty and participated in training 474 Canadian family and general practitioners over 8 months. The learning intervention involved a review of pertinent literature by a local preceptor and a summary of recommendations of the planning committee, followed by demonstration of injector skills and then supervised practice with patients, who received hylan G-F 20 (Synvisc, Ridgefield, NJ) usually in the offices of the family physicians. The pain and function subscales of the WOMAC 3.0 questionnaire were self-administered to each patient in their physician's office, prior to receiving their joint injection and again at or near 6-weeks post-injection. Data were analyzed in the Department of Epidemiology and Biostatistics at The University of Western Ontario, London, ON. RESULTS: Clinically important statistically significant improvements in pain and physical function were noted in patients who received viscosupplementation treatment from family physicians who had recently acquired the necessary injection skills. Approximately three-quarters of the patients experienced a reduction in pain and an improvement in physical function of at least 20%. IMPLICATIONS: These results suggest a positive relationship between acquisition of a new skill by learners and improved patient outcomes as a result of this planned CME learning intervention.

Hemorrhagic ovarian cysts in patients on anticoagulation therapy: CT findings.

OBJECTIVE: Hemorrhage into ovarian cysts is a frequent and potentially life-threatening complication in women on chronic anticoagulation therapy. This article presents the CT findings of three surgically confirmed cases of symptomatic hemorrhagic ovarian cysts associated with anticoagulation therapy. MATERIALS AND METHODS: There were two cases of unruptured hemorrhagic ovarian cysts: a patient with two corpus luteum cysts of menstruation and a patient with a serous cystadenoma. RESULTS: Each mass had a rounded central region of high attenuation (76, 62, and 50 HU) representing blood that was surrounded by fluid density. The third case was a hemorrhagic corpus luteum cyst of menstruation with rupture and hemoperitoneum. CONCLUSION: Hemorrhagic ovarian cysts should be included in the differential diagnosis of high-attenuation adnexal masses, especially in patients on anticoagulation therapy and in those with abrupt onset of pelvic pain.

Role of CT in excluding major arterial injury after blunt thoracic trauma.

The role of CT in the diagnosis of major vascular rupture following blunt decelerating chest trauma is controversial. Its value in excluding major arterial injury has not yet been determined. During a 12-month period we obtained dynamic enhanced thoracic CT studies in 20 patients with blunt decelerating thoracic trauma who had abnormal or equivocal mediastinal contours on chest radiographs. In all cases diagnosis was confirmed by either digital subtraction (18 patients) or conventional thoracic angiography (two patients). CT scans showed evidence of direct aortic injury in three patients and evidence of mediastinal hematoma in five others. Four of these eight patients had major arterial injury verified angiographically and at surgery. In two patients the CT scan was considered equivocal; both patients had normal thoracic angiograms. CT excluded direct vascular injury or mediastinal hematoma in 10 patients. All 10 had normal thoracic angiograms. This preliminary study suggests that, in patients sustaining blunt decelerating thoracic trauma, thoracic CT may be more valuable than chest radiography in excluding major vascular injury and, in some cases, may reduce the need for thoracic angiography.

Value of chest radiography in excluding traumatic aortic rupture.

A retrospective review of chest radiographs from 205 patients with blunt chest trauma who also underwent aortography was performed. Forty-one of the 205 had aortographically proved aortic rupture. Discriminant analysis of 16 radiographic signs indicated that the most discriminating signs were loss of the aorticopulmonary window, abnormality of the aortic arch, rightward tracheal shift, and widening of the left paraspinal line without associated fracture. No single or combination of radiographic signs demonstrated sufficient sensitivity to indicate all cases of traumatic aortic rupture on plain chest radiographs without the performance of a large number of aortographically negative studies. The bedside anteroposterior "erect" view of the chest proved far more valuable than the supine view in detecting true-negative studies. Despite significant reader variability in the interpretation of the various radiographic signs, in general the analysis confirmed the role of chest radiography in this clinical situation, but suggests that its most beneficial use is in excluding the diagnosis and eliminating unwarranted aortography rather than in predicting aortic rupture.

Isolation of human nerve growth factor from placental tissue.

Nerve growth factor (NGF) has been isolated from human placental tissue. Using the chicken embryo dorsal root ganglia assay, we determined levels of NGF activity for the amnion, placental cotyledons, cord serum, fetal serum, and maternal serum. The highest levels of NGF activity were measured in placental cotyledons. After homogenization and centrifugation of the placental cotyledons, the supernatant was sequentially chromatographed, at neutral pH, on Sephadex G-100, DEAE-11, and Sephadex G-150. A high-molecular-weight protein fraction (approximately 150,000), which contained all the biological activity, was isolated in this fashion. Analytical isoelectric focusing of this fraction revealed a basic protein component (pI 9.5) of the high-molecular-weight species. Assays for NGF activity of all protein components separated by analytical isoelectric focusing showed that NGF activity was associated only with the basic protein component. Correspondingly, preparative isoelectric focusing of the high-molecular-weight species yielded a basic protein with very high biological activity (1-3 ng per biological unit) that was immunochemically active against rabbit IgG made against mouse beta-NGF.

The Occurrence and Development of Amylase Enzymes in Incubated, De-embryonated Maize Kernels.

The development of amylase activity in extracts from de-embryonated and GA(3)-treated de-embryonated maize kernels (Zea mays L.) was determined during a 10-day incubation period. The increase in activity was compared with activity extracted from endosperms dissected from germinating whole kernels. Chromatographic analysis of reaction products as well as physicochemical characterization demonstrated that the activities from GA(3)-treated and nontreated tissue were comparable and that part of the activity was attributable to alpha-amylase.Concomitant with the increase in activity was the appearance of a number of starch-degrading bands, as evidenced by polyacrylamide gel electrophoresis. Actinomycin-D (20 mug/ml) and cycloheximide (5 mug/ml), when present in the incubation medium at early periods of incubation, were capable of inhibiting the development of amylase activity and of preventing the appearance of the starch-degrading bands.The results indicate that the development of alpha-amylase activity in de-embryonated maize kernels is independent of an embryo or an exogenous source of gibberellic acid and suggest that this process involved protein synthesis.