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Nonunion remains a major complication after skeletal trauma. In the last decade, extracorporeal shock wave therapy has become a common tool for the treatment of nonunions. To date, no prospective, randomized trial has been conducted to show the efficacy of this form of treatment.This study was performed to determine the value of extracorporeal shock wave therapy for nonunions. Previous published results in the literature and our own clinical results were analyzed and related to the natural history of bony union.No study has proven that extracorporeal shock wave therapy improves bone healing. Clinical studies reporting the acceleration of union after application of shock waves instead seem to misinterpret the natural history of bony union.No evidence supports the treatment of pseudarthroses with extracorporeal shock waves. A randomized, prospective, clinical trial with a control group has to be performed before a final decision can be made regarding this indication for extracorporeal shock wave therapy.

Instruments used in surgery which rotate or vibrate at a high frequency can produce potentially contaminated aerosols. Such tools are in use in cemented hip revision arthroplasties. We aimed to measure the extent of the environmental and body contamination caused by an ultrasound device and a high-speed cutter. On a human cadaver we carried out a complete surgical procedure including draping and simulated blood flow contaminated with Staphylococcus aureus (ATCC 12600). After cemented total hip arthroplasty, we undertook repeated extractions of cement using either an ultrasound device or a high-speed cutter. Surveillance cultures detected any environmental and body contamination of the surgical team. Environmental contamination was present in an area of 6 x 8 m for both devices. The concentration of contamination was lower for the ultrasound device. Both the ultrasound and the high-speed cutter contaminated all members of the surgical team. The devices tested produced aerosols which covered the whole operating theatre and all personnel present during the procedure. In contaminated and infected patients, infectious agents may be present in these aerosols. We therefore recommend the introduction of effective measures to control infection and thorough disinfection of the operating theatre after such procedures.

Most patients depend on glasses for near vision after a cataract surgery with implantation of a monofocal IOL. Implantation of a multifocal IOL offers these patients the possibility to become independent of glasses. The aim of this study was to gain clinical experience with the multifocal MF4-Lens (IOLtech, France).The MF4-lens was implanted in 80 eyes of 40 patients. Corrected and uncorrected near and far vision acuity were evaluated 3 months postop. Patients' satisfaction, the appearance of disturbing optical phenomena and the dependence on glasses were ascertained by means of an anonymous questionnaire.No surgical complications occurred in the course of this study. Three months after surgery, 92 % of all patients had an uncorrected visual acuity of 0.5 (20/40) or better. The binocular, uncorrected visual acuity was 0.63 (20/32) or better. The corrected far-vision acuity was 0.5 (20/40) or better. 86 % of all patients had an uncorrected near-vision acuity of 0.8 (20/25) and 45 % an uncorrected near-vision acuity of 1.0 (20/20). 94 % of the patients were satisfied with the results of the surgery. 38 % reported on glare and 45 % noticed halos. 67 % of all patients were totally independent of glasses, 27 % needed glasses frequently and 6 % generally.The implantation of the multifocal MF4-lens is an effective procedure in particular for near-vision acuity and patient satisfaction. The majority of patients were less dependent on glasses.

In 2009 the Swiss Federal Laboratories for Materials Testing and Research (EMPA) and the Paul Scherrer Institute (PSI) developed a hydrogen-powered municipal street cleaning vehicle, which has been in real-world operation in Basel. The vehicle has now been taken to St Gallen in northeastern Switzerland for further practical trials.

Study design The assignment of idiopathic scoliosis curves to the curve types,1-6 to the lumbar spine modifier (A, B, or C), and to the sagittal thoracic modifier (-, N, +), as recently described by Lenke et al, was evaluated by five observers on two occasions. Objective To determine the intraobserver and interobserver reliabilities of Lenke's new system for classifying idiopathic scoliosis. Summary of background data Lenke et al recently introduced a new system for classifying idiopathic scoliosis. Methods Preoperative standing long cassette coronal and sagittal radiographs as well as side-bending radiographs of 51 consecutive patients who underwent surgery for idiopathic scoliosis were labeled with the Cobb angles for all the curves. The center sacral vertical line was marked as the bisection of the proximal sacrum perpendicular to the true horizontal line. Five observers independently assigned a curve type, a lumbar spine modifier, and a sagittal thoracic modifier to each curve following the guidelines as described by Lenke et al. Assignment of the curves was repeated 3 weeks later, with the curves presented in a different order. Kappa coefficients were used to determine the interobserver and intraobserver reliabilities. Results All five reviewers agreed on the overall classification in 21 (41%) of the 51 patients. A mean kappa value of 0.62 was determined for interobserver reliability, and a mean kappa of 0.73 for intraobserver reliability. Determination of an upper thoracic curve as structural or nonstructural and assignment of a lumbar spine modifier were the main reasons for disagreement. Conclusions Lenke's new classification system is more reliable than the older King classification, but proper classification of high thoracic and lumbar curves seems to be difficult.

Surgery is no longer what we knew it to be. Rapid technological and medical developments, and especially a changing infrastructural, political and ethical environment, have changed the face of modern surgery. This puts more and novel demands on the surgeons and the surgeons-to-be. When we were asked by Professor Shokeir to put together another special issue for the AJU, being endourologists what first came to our minds were all sorts of sub-speciality issues. Then we asked what could connect all urologists worldwide. Not only that, but what connects them with the rest of the surgical community, and with associated co-operating specialties? The answer is simple: training. The training to do the right thing for the right patient at the right time, in the context of urology, surgery, and medicine in general. All of us are in training. Training does not stop with graduation, or even a professorship. Training and learning is a life-long process. Some still learn when they look immediate death in the eye. Trainees need to learn how to master the skills required, and then how to become teachers themselves. Teachers have to learn how to become better teachers, not least by listening to the trainees. In surgery, over centuries the training and teaching followed the apprenticeship model. However, the increasing accountability of surgeons, increasing patient awareness, and reductions in working times for surgeons and trainees in an attempt to achieve a safer work environment and a better work-life balance, have made this model unsuitable for modern training and teaching in many parts of the world. In addition, new learning concepts are continuously evolving and often cross traditional borders between trainer and trainee, surgery and other medical specialties, and even extra-medical professions. What has been good for centuries does not necessarily mean that it cannot be improved. Trainers and trainees can learn from each other, as can surgeons from anaesthesiologists, or even from other high-risk professions such as airline pilots. In this issue we tried to bring together all these aspects of training and teaching, from concepts to practice, from the trainees view to the trainers view, from the educationalist to the practical urologist. We believe that this special issue might not only be of interest to trainers, but for all of us, as we all keep learning and we all should be keen to understand how we do it, how others do it and how we can improve our current beliefs and standards.

Study Design. A cadaver study to evaluate contamination in the operating room through the use of a high-speed bone cutter. Objectives. To determine the grade of contamination of animate and inanimate objects through an aerosol intraoperatively, produced by a high-speed cutter during lumbar laminectomy. Summary of Background. In spinal surgery, high-speed cutters are used that produce an aerosol consisting of a mixture of irrigation solution, blood, and tissue debris. Such aerosols can be contaminated with potential pathogens. The surgical personnel and the environment are therefore exposed to a contamination risk. Methods. Laminectomies at three points (L2–L4) were performed on a human cadaver using a high-speed cutting device. The aerosol produced by the irrigation solution was contaminated with Staphylococcus aureus ATCC 12600. To detect the contamination of the environment and of the surgical team, surveillance cultures were used. Results. By air sampling, staphylococci were detected in the operating room at an extension of 5 by 7 m. The surgical team showed extensive face and body contamination with S. aureus. Despite protection by a barrier drape, similar contamination was observed on both the cadaver’s head and the anesthesiologist. Conclusions. The use of high-speed cutters in spinal surgery produces an aerosol that can be contaminated with blood-borne pathogens from infected patients. This aerosol is spread over the whole surgical room and contaminates the room and all personnel present. It is therefore critical to ensure that effective infection control measures are performed, not only by the surgeons but by everyone present in the operating room. The room itself must be sufficiently disinfected after such procedures.

A follow-up of more than 10 years among patients who have undergone a total knee arthroplasty (TKA) was performed to determine the significance of patella resurfacing for the long-term outcome. The clinical outcome was assessed by the Knee Society Score (KSS), and the radiological outcome was determined based on the Knee Society Roentgenographic Evaluation System. The patella was preserved in 21 knees and resurfaced in 44 knees. The mean follow-up time was 11.6 years (range 10-16.3 years). There was no significant difference in the clinical outcome between the knees with patella resurfacing (knee points: mean 85.3 +/-12.9, function points: mean 70.3 +/- 23.4) and the knees with patella retention (knee points: mean 82.7 +/- 16.2, function points: mean 71.7 +/- 22.4; p = 0.58 for knee, and p = 0.83 for function points). There was also no significant difference in the radiological outcomes regarding the angles alpha, beta, Upsilon, delta, and valgus ( p0.05 for each variable). There was, however, a trend towards more lucencies in TKAs with a resurfaced patella on the tibia side in the anteroposterior view ( p = 0.052). Patellar complications were found more often in the resurfaced group (20.5%) than in the group without resurfacing (9.6%). The results indicate overall no advantage of patella resurfacing compared with patella retention in the long run.

For solving complex analytical tasks networking has proven to be extremely useful. Networking in Analytical Chemistry shows various aspects. The use of a pool of different, complementary instrumental methods for ambitious analytical investigations is fundamental. Furthermore, stimulating a transdisciplinary approach through dynamic team formation within a transparent, flexible organization is an essential management task. Two case studies illustrating the networking approach at EMPA in complex analytical service and R & D projects are presented. The result is a comprehensive approach in analytical chemistry, activating synergy.

Pre-mRNA processing, referred to as alternative RNA splicing (AS), is a critical determinant of protein diversity. AS produces multiple transcripts and, as a result, multiple proteins from a single gene. Recently, frequent mutations in splicing factor genes have been reported in myelodysplasia (MDS) and chronic lymphocytic leukemia (CLL), and less frequently in AML. However, in previous studies, we found that aberrant patterns of splicing were common in cells from 66 AML patients compared to 10 normal donors (NDs), more common than could be explained by mutations in splicing factor genes. Here, we evaluated expression levels of 24 core splicing factors (SFs), which are involved in splicing reactions, in cells from 30 AML patients compared to 10 NDs. Among these SFs we identified three, U2AF2, PTBP, and SFRS12 that were significantly (P Disclosures: Griffin: Novartis: Research Funding; Janssen: Research Funding.

Background The presence of abundant macrophages is characteristic morphological hallmark for the diagnosis of Burkitt´s lymphoma. They are considered to be the removers of cellular debris which occurs due to the rapid proliferation of such lymphomas. However, although macrophages represent the main non-malignant population within such lymphomas and therefore can be considered a substantial part of the stroma, little is known about how these cells are being recruited and how they influence malignant growth. Since malignant B–cells show high plasticity and have capacity to differentiate into various phenotypes we analysed the origin of macrophages in a murine model of Burkitt´s lymphoma. In this transgenic model a human c-myc gene is overexpressed in a B-cell specific fashion. Results When long term cultured lymphoma cells from transgenic animals were transferred into wild type animals expressing the congenic marker CD45.1 lymphomas grew out within 14 days post transfer. Lymphomas were minced into single cell suspension and macrophages were sorted by flow cytometry based on the expression of CD11b and F4/80. Approximately 6% of the cells expressed F4/80 and CD11b as macrophage markers and displayed an M2 phenotype. Surprisingly approximately 50% of these macrophages also expressed CD45.2 and therefore were not of host origin but derived from the transferred lymphoma cells. These cells were positive for the c-myc transgene, but displayed reduced expression of c-myc as shown by RT PCR in comparsion to the lymphoma cell line in culture. Furthermore, lymphoma derived macrophages displayed reduced PAX5 expression and were negative for CD19 surface expression. When lymphoma cells were retrovirally transduced to express bcl2 to block apoptosis, and were treated with chemotherapy in vivo, we observed a massive increase in macrophage numbers in remaining tissues suggesting transdifferenziation as an alternative exit strategy in case of cellular stress. Conclusion These results demonstrate that BL cells are able to differentiate into a macrophage phenotype to form their own supporting stroma. In addition, differentiation after DNA damage such as chemotherapy is increased suggesting a new exit strategy when classic apoptotic pathways are blocked. Disclosures: Mackensen: Eli Lilly: Consultancy.

Most schools of engineering and business today have not yet come to terms with the growing inter-disciplinary education needs that adequately prepare PhD graduates for assuming leadership, professional, academic and/or research positions in academia, industry or government. The purpose of this paper is to contribute to a better understanding of how to develop an innovative inter-disciplinary PhD degree programme by presenting a case study in technology management at the University of Bridgeport. The proposed PhD programme is currently undergoing an external review process. The paper will describe the purpose, structure, and approach of the proposed PhD programme as well as the challenges, lessons learned and critical success factors.

Based on our current knowledge, particulate matter in Diesel-engine exhaust gases should be classified as toxic. The current regulations define limits only for the total mass emission of particulate matter given in g/km or per energy unit. Other properties such as particle size or chemical composition are not yet considered.The EMPA has been concerned with the detailed analysis of particulate emission since 1992. For this investigation, samples of Diesel exhaust gases and heating-system flue gases are being used. The Diesel engine is operating under various conditions whereas two different oil-burner types are being used in the heating system. The burners are designated 'yellow-flame' and 'blue-flame'.The size distribution of the particles, their morphology, and chemical composition are determined. The initial results show that the particle properties depend on the combustion processes and operating conditions. This investigation is being continued.

Abstract 3491 Novel methods to quantify chimerism after allogeneic hematopoietic stem cell transplantation (HSCT) based on single-nucleotide polymorphism (SNP) specific real-time quantitative polymerase chain reaction (qPCR) require high amounts of input DNA. The applicability of these methods in cases where DNA quantity is limiting, however, is currently unclear. Here, we demonstrate that SNP typing performed with just 5ng of input DNA still allowed for the distinction of positive (mean ct 28.05) and negative (ct > 36) signals. In addition, we confirm the high informative value of a set of 19 SNP markers, with 12 markers exceeding 20% informativity in our population (n=74). Of interest, a four-fold reduction of input DNA for qPCR standard curves did not alter PCR efficiencies. Most importantly, in 7 out of 16 allogeneic HSCT patients who experienced disease relapse, a retrospective analysis using the SNP-qPCR method revealed re-appearance of recipient chimerism earlier (mean 95 days) compared to previous results obtained by a short tandem repeat (STR) specific PCR. Taken together our data clearly demonstrate that SNP-qPCR is more sensitive compared to the widely used STR-PCR even with reduced amounts of input DNA. We therefore recommend that SNP-qPCR is preferable to STR-PCR for chimerism analysis in cases where DNA quantity is a limiting factor. Disclosures: No relevant conflicts of interest to declare.

The metal free apoprotein of recombinant human transcription factor SP1 was used in metal reconstitution experiments to study the importance of zinc in facilitating DNA binding of zinc finger proteins. Our functional analysis indicates that several transition metals are capable of modulating DNA-protein interactions of zinc finger domains with their cognate DNA target sites. Excess or deficiency of divalent zinc, or the presence of transition metals, such as divalent cadmium, cobalt, copper, manganese and nickel impair DNA binding of zinc reconstituted SP1. In addition, functionally active SP1 protein can be obtained by metal reconstitutions in absence of zinc(II) by presence of cadmium(II)- and cobalt(II)-, to lesser extents by presence of nickel(II)- or manganese(II)chloride. This study indicates that zinc might play a functional role in regulating DNA protein interactions of zinc finger proteins in vivo. It is postulated that fluctuating divalent zinc alone or transition metals bound to cellular components might form mixed-ligand complexes that alter the zinc finger protein conformation and impair DNA binding.

Lumbar disc arthroplasty has become a popular modality for the treatment of degenerative disc disease. The dimensions of the implants are based on early published geometrical measurements of vertebrae; the majority of these were cadaver studies. The fit of the prosthesis in the intervertebral space is of utmost importance. An undersized implant may lead to subsidence, loosening and biomechanical failure due to an incorrect center of rotation. The aim of the present study was to measure the dimensions of lumbar vertebrae based on CT scans and assess the accuracy of match in currently available lumbar disc prostheses. A total of 240 endplates of 120 vertebrae were included in the study. The sagittal and mediolateral diameter of the upper and lower endplates were measured using a digital measuring system. For the levels L4/L5 and L5/S1, an inappropriate size match was noted in 98.8% (Prodisc L) and 97.6% (Charite) with regard to the anteroposterior diameter. Mismatch in the anterior mediolateral diameter was noted in 79.3% (Prodisc L) and 51.2% (Charite) while mismatch in the posterior mediolateral diameter was observed in 91.5% (Prodisc L) and 78% (Charite) of the endplates. Surgeons and manufacturers should be aware of the size mismatch of currently available lumbar disc prostheses, which may endanger the safety and efficacy of the procedure. Larger footprints of currently available total disc arthroplasties are required.

The mixed lineage leukemia (MLL) family of histone methyltransferases has become notorious for the participation of the founding member MLL in fusion proteins that cause acute leukemia. Despite a high overall structural conservation no other MLL homolog has been found involved in leukemogenic fusion proteins. This surprising fact might be either due to a relative genomic stability of the MLL2 locus that prevents chromosomal translocations or MLL2 might be altogether incapable of contributing to oncogenic fusions because of functional constraints. Here we demonstrate the latter to be true for fusion proteins based on MLL2, the closest relative of MLL. A MLL2-ENL protein constructed in analogy to the highly leukemogenic MLL-ENL was incapable of transforming primary hematopoietic cells. Elaborate swap experiments identified the zinc binding “CxxC”- region of MLL2 and an adjacent “post-CxxC” stretch of basic amino acids as the essential determinants for the observed difference between MLL and MLL2. Surprisingly gel shift experiments indicated that the CxxC and post-CxxC domains of MLL and MLL2 conferred almost indistinguishable in vitro DNA binding properties. However, in vivo these motifs guided MLL-ENL and MLL2-ENL to a largely non-overlapping gene repertoire. Divergent nuclear localization, a distinct potency to activate a MLL model promoter in reporter assays, and measurements of homeobox gene levels in primary cells expressing MLL and MLL2 fusion proteins all argued for a separate target site selection of the two fusions. Therefore, the CxxC domain appears to be largely responsible for target finding and as a consequence this domain is a promising object for therapies aimed at MLL fusion proteins without affecting the general function of other MLL family members.

Chromosomal translocations at 11q23 are characteristic for a subset of particularly aggressive acute lymphoid and myeloid leukemias. This event fuses the mixed lineage leukemia (MLL) gene to a puzzling variety of seemingly unrelated fusion partners to create novel transcription factors with potent oncogenic activity. Despite the molecular heterogeneity, MLL−associated leukemias show similar clinical presentations and an almost identical gene expression pattern regardless of the translocation partner. We and others previously identified interactions between individual MLL translocation partners suggesting that these might share a common mechanism for transformation. To characterize these interactions in more detail we generated a HEK293−derived producer line expressing a FLAG−tagged derivative of the frequent fusion partner ENL. Double affinity purification of nuclear extracts on FLAG and ENL antibody resins reproducibly yielded a macromolecular complex of 10 proteins we term MPAC, for MLL Partner Associated Complex, which in addition to ENL includes the MLL fusion partners AF4, AF9 and AF5q31, and two components involved in transcriptional elongation: pTEFb and DOT1L. The pTEFb dimer consists of cyclin T1/2 and CDK9 and is responsible for serine−2 phosphorylation of the C−terminal repeat domain of elongating RNA Polymerase II. DOT1L is the only known histone methyltransferase that catalyzes dimethylation of H3K79, a modification introduced during elongation of actively transcribed transcription units. Surprisingly, PC3 and RING1b, members of the polycomb repressive complex I were also identified in the MPAC complex. Further examination demonstrated a direct interaction of ENL with DOT1L through C−terminal interaction domains by yeast 2−hybrid, pulldown and colocalization experiments. Knock−down of ENL diminished global H3K79 dimethylation and also impaired various promoters in transient reporter assays suggesting the MPAC complex is involved in transcriptional regulation mediated by DOT1L, even in the absence of MLL fusion proteins. In summary, our results indicate that the most common MLL fusion partners are associated in an hitherto undescribed complex involved in transcriptional elongation.

Overexpression of Hox genes is frequently associated with leukemogenic transformation of hematopoietic cells. In particular the abdominal-type HoxA genes together with their homeodomain containing dimerization partners of the TALE (three amino acid loop extension) family are well documented oncogenes in various hematologic malignancies. Paradigmatic examples are Hoxa9 and Meis1 that are coactivated by retroviral insertions in the BXH2 mouse leukemia model. In addition Hoxa9 is a fusion partner of NUP98 in acute myeloid leukemias with a t(7;11) and Meis1 as well as Hoxa9 are critical targets of MLL (mixed lineage leukemia) fusion proteins. Despite their important role in leukemogenesis little is known how they exert their transforming function. Here we used a novel system of transient transformation by an inducible oncoprotein to generate populations of primary hematopoietic cells that overexpress Hoxa9 in combination with Meis1. The RNA inventory of these cells was compared to control cells by Affymetrix array analysis. Interestingly the RNA of c-Myb (myeloblastosis leukemia virus oncogene) and of a known c-myb downstream target (Gstm1) was amongst the top ten genes upregulated by Hoxa9/Meis1. c-Myb overexpression was verified by Northern blot and quantitative RT-PCR. c-Myb levels also responded to the introduction of MLL-ENL, an oncoprotein that upregulates Hoxa9 and Meis1. Small interfering RNAs directed against c-Myb suppressed transformation by MLL-ENL but had no effect on transformation of hematopoietic precursors by E2A-HLF an oncogene that does not work by increasing Hoxa9/Meis1 levels. The anti c-Myb siRNA effect was abrogated by coexpression of a c-Myb derivative with a mutated siRNA target site. The coexpression of a dominant negative c-Myb mutant had a similar but weaker effect on MLL-ENL mediated transformation. Ectopic expression of c-Myb in hematopoietic precursor cells induced a mild differentiation block. However, c-Myb alone was not able to give a positive readout in a serial replating assay under conditions where the combined overexpression of Hoxa9 together with Meis1 was clearly transforming. In summary the results suggest that c-Myb is essential but not sufficient for the oncogenic activity of Hoxa9/Meis1.

Time-resolved Chemical Ionisation Mass Spectrometry (CI-MS) has been used to investigate the benzene emission profiles in automotive exhaust during transient engine operation. Representative petrol-driven passenger cars and light duty vehicles with model years 1991 to 1996 have been selected from the Swiss car fleet and were measured on the chassis dynamometer at the Swiss Federal Laboratories for Materials Testing and Research (EMPA). The benzene emission characteristics of a fleet of 20 passenger cars (1.3-5.2 l), eight light duty vehicles (2.0-2.4 l) and one conventional vehicle (1.4 l) have been determined. A set of seven driving cycles, including the European Driving Cycle (ECD), the US Urban (FTP 75), the Highway and four additional driving cycles have been investigated by CI-MS. Velocity-dependent emission factors for the individual vehicles as well as for the different vehicle classes were determined by a statistical analysis of the corresponding emission data.

Instantaneous emissions measurements on chassis dynamometers and engine test benches are becoming increasingly normal for environmental emission factor calculation. Modern exhaust gas analysers allow measurement at a rate of one sample per second, thus creating the impression that the emissions information recorded reflects that precision. Normally, the time delay between the car and the analysers is considered to be constant and compensated for accordingly. In reality, this time delay varies by more than three seconds depending on the engine load. Moreover, emission peaks are smoothed by turbulence during transport. The analyser's dynamics smooth the signals even more. For example, an emission peak of one second is flattened to a three-second peak in raw gas measurement, and to a very low hill of about ten seconds in diluted gas measurement. If emissions measurements are to be correlated to actual driving conditions, the variable time delays and the smoothing must be compensated for. To achieve this, the time delay at a given time can be calculated by dividing the volume of the exhaust system by the actual exhaust flow. "Smoothing" is compensated for by adopting a system theory-based approach. It can be described with a linear differential equation, which can be "inverted" in an offline procedure.