Nodes-and-connections RNAi knockdown screening: identification of a signaling molecule network involved in fulvestrant action and breast cancer prognosis.

Although RNA interference (RNAi) knockdown screening of cancer cell cultures is an effective approach to predict drug targets or therapeutic/prognostic biomarkers, interactions among identified targets often remain obscure. Here, we introduce the nodes-and-connections RNAi knockdown screening that generates a map of target interactions through systematic iterations of in silico prediction of targets and their experimental validation. An initial RNAi knockdown screening of MCF-7 human breast cancer cells targeting 6560 proteins identified four signaling molecules required for their fulvestrant-induced apoptosis. Signaling molecules physically or functionally interacting with these four primary node targets were computationally predicted and experimentally validated, resulting in identification of four second-generation nodes. Three rounds of further iterations of the prediction-validation cycle generated third, fourth and fifth generation of nodes, completing a 19-node interaction map that contained three predicted nodes but without experimental validation because of technical limitations. The interaction map involved all three members of the death-associated protein kinases (DAPKs) as well as their upstream and downstream signaling molecules (calmodulins and myosin light chain kinases), suggesting that DAPKs play critical roles in the cytocidal action of fulvestrant. The in silico Kaplan-Meier analysis of previously reported human breast cancer cohorts demonstrated significant prognostic predictive power for five of the experimentally validated nodes and for three of the prediction-only nodes. Immunohistochemical studies on the expression of 10 nodal proteins in human breast cancer tissues not only supported their prognostic prediction power but also provided statistically significant evidence of their synchronized expression, implying functional interactions among these nodal proteins. Thus, the Nodes-and-Connections approach to RNAi knockdown screening yields biologically meaningful outcomes by taking advantage of the existing knowledge of the physical and functional interactions between the predicted target genes. The resulting interaction maps provide useful information on signaling pathways cooperatively involved in clinically important features of the malignant cells, such as drug resistance.

[Paraparesis following stable L4 burst fracture Possible correlation with preexisting degenerative stenosis of the spinal canal]. / Paraparese nach stabiler LWK-4-Fraktur Möglicher Zusammenhang mit vorbestehender degenerativer Spinalkanalstenose.

A patient with degenerative stenosis of the lumbar spinal canal developed paraparesis of both legs following a stable L4 burst fracture. However, neither the stenosis nor the fracture have caused a significant narrowing of the spinal canal. Surgical decompression and dorsal stabilization has led to relief of the neurological deficit. This case demonstrates that even a stable burst fracture may cause paraparesis and that a degenerative spinal canal stenosis is a predisposing factor.

[Prevalence of rotator cuff defects in dislocated fractures of the humerus head in elderly patients]. / Die Prävalenz von Rotatorenmanschettendefekten bei der dislozierten Humeruskopffraktur des älteren Patienten.

Humeral head fractures are commonly found in elderly patients. A high rate of spontaneous rotator cuff tears in this patient group may endanger good results following surgical treatment. 70 patients > 65 years with dislocated humeral head fractures were prospectively studied and treated surgically following a standardised treatment protocol. In all cases the rotator cuff was examined intraoperatively. A tear of the rotator cuff was documented in 4 cases (5%). The prevalence of rotator cuff tears in elderly patients with dislocated humeral head fractures is significantly smaller than expected referring to magnetic resonance-, sonographic and cadaveric studies of same-age patients without fracture. Therefore the rotator cuff may serve as retention anchor for the surgical treatment of dislocated humeral head fractures in elderly patients.

[Ultrasound imaging of the ventrocaudal labrum in ventral instability of the shoulder]. / Die sonographische Darstellung des ventrokaudalen Labrums bei der ventralen Instabilität der Schulter.

The significance of the anterior capsulolabral complex in anterior shoulder instability is well established. In cases of detachment of the ventrocaudal capsulolabral complex, definitive stability can be achieved only by operative refixation. These structures can only be imaged by invasive and cost-intensive methods such as MR arthrography and CT arthrography. We introduce an examination by ultrasonography: a 7.5 MHz linear transducer is used. The patient is in the supine position and both arms are abducted and externally rotated. The transducer is placed in the longitudinal direction, parallel to the border of the pectoral muscle with a ventrocaudal tilt. From March 1992 to October 1994 a number of 88 patients were preoperatively subjected to ultrasound examination before operative stabilization or arthroscopy. Eighty-three of the results were evaluable. Sixty-nine of the examinations showed evidence of detachment of the ligaments; 68 of them were confirmed operatively; 14 of the examinations showed no evidence of detachment; 12 of these results proved true operatively (sensitivity: 97.7%, specificity 92.3%). The sonographic procedure introduced seems to be valuable in detecting the Bankart lesion in anterior instability of the shoulder.

[Value of sonography of the anterior cruciate ligament in post-traumatic hemarthrosis]. / Die Aussagekraft der Sonographie des vorderen Kreuzbandes beim posttraumatischen Hämarthros.

Periosteal avulsions of the femoral insertion of the anterior cruciate ligament (ACL) should be treated by reinsertion within 2 weeks of the injury. Due to painful muscular hypertension, the clinical examination of ligament stability incorrectly revealed negative results in 12-62% posttraumatically. The only sufficiently sensitive indication of ACL rupture is posttraumatic hemarthrosis. Therefore, early arthroscopy of any posttraumatic hemarthrotic knee joint is required to exclude or confirm ACL rupture. Up to 30% of such procedures are carried out with no clinical consequences. We therefore set out to test the value of sonography of the ACL in hemarthrosis of the injured knee joint. A total of 117 posttraumatic hemarthrotic knee joints were subjected to ultrasound examination prior to arthroscopic evaluation. With a 5 MHz linear scanner, the femoral insertion of the ACL at the femoral condyle is represented in the same projection of injured and noninjured knee joints. In noninjured knee joints the hypoechoic insertion of the ACL lies directly on the S-shaped line of the lateral femoral condyle. The space between the condyles is hyperechoic. In case of a femoral ACL avulsion the insertion of the ACL and the hyperechoic structures between the condyles are forced aside by a hypoechoic area. Of 51 arthroscopically demonstrated complete ruptures of the ACL, 49 were detected by sonography. In 16 cases sonography suggested ACL rupture, but this could not be confirmed by arthroscopy, though 8 of these cases showed evidence of ACL strain without any lost of stability of the knee joint. For sonography we found a sensitivity of 96.1% and a fairly good specificity of 75.2%.(ABSTRACT TRUNCATED AT 250 WORDS)

[Arthroscopy following pediatric injuries of the knee joint--a helpful or superfluous diagnostic procedure?]. / Die Arthroskopie nach kindlichen Kniegelenkverletzungen--ein hilfreiches oder überflüssiges diagnostisches Verfahren?

Between 1983 and 1987, 1082 arthroscopies were carried out on the knee joint. This included 48 cases of children between the age of seven and fifteen with knee injuries. In 21 cases the clinical diagnosis was confirmed. In 5 cases the clinically indications for an operation could not be confirmed. In 9 cases of arthroscopy, it was demonstrated that an operation was necessary, although this was not clinically diagnosed. Finally in 15 cases, specific conservative therapy could be planned on the basis of the medical evidence. From 35 cases of haemarthrosis, 27 proved to have injuries requiring therapy, 12 of which were not obvious during clinical examination. The results underline advantages of arthroscopy for childhood knee joint injuries, in allowing a definite diagnosis and varying therapeutic measures.

Autogenous fibula grafting of a radial defect complicating acute hematogenous osteomyelitis in a child.

The emphasis of this case report is on the management of a radial defect after diaphysectomy complicating acute hematogenous osteomyelitis in a 15-month-old girl. An autogenous fibular graft was used to replace the defect of the radius. It was wedged between the proximal and distal radial fragment and fixed in position by two miniplates. After 6 weeks the fibula had incorporated. The graft fractured, unfortunately, through a drill-hole created for neovascularization, requiring further interventions until complete cure occurred. A re-examination 2 years after grafting revealed a good functional result. Radiographically, the radius was completely remodeled. We consider the autogenous fibular graft to be especially suitable for replacing the radius for the following reasons: it is firm enough for bridging, it has a corresponding caliber, and it can be obtained with adequate length. Within a few weeks the fibular defect refills with new bone formation, provided that the periosteum has been tightly sutured. No interference with ankle growth was observed.

Quantification of antimyosin antibodies in experimental myocarditis by a new solid phase fluorometric assay.

A solid phase fluorometric assay of high sensitivity and specificity for detection of antimyosin antibodies is described. The method consists of 4 basic steps: (1) coating the carrier stiQ with myosin; (2) incubation of the stiQ in the test serum; (3) labeling of the bound antibodies by FITC-anti-immunoglobulin antibodies; and, (4) quantitation of the bound antibodies in a fluorometer (FIAXTM). Using a purified myosin preparation as antigen, the assay was tested in experimental myocarditis of rabbits injected with cardiac proteins. Reproducibility was excellent, standard deviation of intra-assay variance being 7.86%, and inter-assay variance 9.45%. When compared with immunodiffusion, a complement fixation test with myosin and indirect immunofluorescence, the assay described proved superior since it allows simple monitoring of serum concentrations of antimyosin antibodies.