[Physical treatment options with impact on bone healing]. / Physikalische Verfahren mit Einfluss auf die Knochenheilung.

The impact of physical stimulation of a fracture remains unsolved because of the complexity of this process. Differences in the localization and the morphology of the fracture, soft tissue injury, pretreatment and risk factors have an influence on study results, leading to problems in evaluation of physical modulation concerning fractures and nonunions. Extracorporeal shock wave therapy (ESWT) is technically demanding and often associated with local complications including bone and soft tissue stress; however, it is still applied in some centers for the treatment of nonunions. The study situation assessing the effectiveness of this treatment consists of a single randomized controlled trial (RCT) with a medium risk of bias. A positive effect for bony healing could be seen in 70-71% of ESWT patients but also in 73% of the surgically treated group. A systematic review and meta-analysis demonstrated an insufficient and inconsistent study quality but acknowledged that ESWT can be an effective treatment for delayed union and nonunion. For low-intensity pulsed ultrasound (LIPUS) studies with a reduced bias are available, representing a better level of evidence. Concerning advantages in the consolidation of acute fractures, only one RCT showed a reduction of healing time in fractures with an expected prolonged bony bridging. For the treatment of delayed unions, a highly rated RCT showed a significantly improved consolidation of midshaft tibial fractures using LIPUS. A systematic review and meta-analysis of nonunions showed positive effects in biologically active lesions, e.g. in hypertrophic pseudarthrosis, leading to a fusion rate of 80%. The consolidation process was better in patients without surgical revision 3-6 months prior to LIPUS.

Distribution of soluble and microsomal epoxide hydrolase in the mouse brain and its contribution to cerebral epoxyeicosatrienoic acid metabolism.

Epoxide hydrolases comprise a family of enzymes important in detoxification and conversion of lipid signaling molecules, namely epoxyeicosatrienoic acids (EETs), to their supposedly less active form, dihydroxyeicosatrienoic acids (DHETs). EETs control cerebral blood flow, exert analgesic, anti-inflammatory and angiogenic effects and protect against ischemia. Although the role of soluble epoxide hydrolase (sEH) in EET metabolism is well established, knowledge on its detailed distribution in rodent brain is rather limited. Here, we analyzed the expression pattern of sEH and of another important member of the EH family, microsomal epoxide hydrolase (mEH), in mouse brain by immunohistochemistry. To investigate the functional relevance of these enzymes in brain, we explored their individual contribution to EET metabolism in acutely isolated brain cells from respective EH -/- mice and wild type littermates by mass spectrometry. We find sEH immunoreactivity almost exclusively in astrocytes throughout the brain, except in the central amygdala, where neurons are also positive for sEH. mEH immunoreactivity is abundant in brain vascular cells (endothelial and smooth muscle cells) and in choroid plexus epithelial cells. In addition, mEH immunoreactivity is present in specific neuronal populations of the hippocampus, striatum, amygdala, and cerebellum, as well as in a fraction of astrocytes. In freshly isolated cells from hippocampus, where both enzymes are expressed, sEH mediates the bulk of EET metabolism. Yet we observe a significant contribution of mEH, pointing to a novel role of this enzyme in the regulation of physiological processes. Furthermore, our findings indicate the presence of additional, hitherto unknown cerebral epoxide hydrolases. Taken together, cerebral EET metabolism is driven by several epoxide hydrolases, a fact important in view of the present targeting of sEH as a potential therapeutic target. Our findings suggest that these different enzymes have individual, possibly quite distinct roles in brain function and cerebral EET metabolism.

Pharmacogenetics of modafinil after sleep loss: catechol-O-methyltransferase genotype modulates waking functions but not recovery sleep.

Sleep loss impairs waking functions and is homeostatically compensated in recovery sleep. The mechanisms underlying the consequences of prolonged wakefulness are unknown. The stimulant modafinil may promote primarily dopaminergic neurotransmission. Catechol-O-methyltransferase (COMT) catalyzes the breakdown of cerebral dopamine. A functional Val158Met polymorphism reduces COMT activity, and Val/Val homozygous individuals presumably have lower dopaminergic signaling in the prefrontal cortex than do Met/Met homozygotes. We quantified the contribution of this polymorphism to the effects of sleep deprivation and modafinil on subjective state, cognitive performance, and recovery sleep in healthy volunteers. Two-time 100 mg modafinil potently improved vigor and well-being, and maintained baseline performance with respect to executive functioning and vigilant attention throughout sleep deprivation in Val/Val genotype subjects but was hardly effective in subjects with the Met/Met genotype. Neither modafinil nor the Val158Met polymorphism affected distinct markers of sleep homeostasis in recovery sleep. In conclusion, dopaminergic mechanisms contribute to impaired waking functions after sleep loss.

[Clinical accuracy of fluoroscopic navigation at the thoracic and lumbar spine]. / Klinische Präzision der C-Arm-Navigation an der Brust- und Lendenwirbelsäule.

The aim of the present study was the verification of the accuracy of 2D fluoroscopy-based navigated pedicle screw placements at the thoracic and lumbar spine in a case series of traumatised patients. Within 36 months 111 pedicle screws were instrumented using C-arm based navigation in 29 patients, 60 at the thoracic and 51 at the lumbar spine. All screw positions were evaluated postoperatively by a routine thin-slice CT scan using multiplanar reconstruction. The position of a screw in relation of its pedicle was classified as: a) screw completely intraosseous, b) screw perforated less than thread level and c) screw perforated over thread level. In 34 thoracic (56.7%) and 32 lumbar (62.7%) screws complete intraosseous placement was observed, 14 thoracic screws (23.3%) and 14 lumbar screws (27.5%) perforated less than thread level. Perforations over thread level were found in 12 thoracic (20%) and 5 lumbar (9.8%) screws. Only medial and lateral perforations of the pedicle were documented (without neurological signs), cranial or caudal perforations did not occur. Segmentation of the C-arm navigation into two comparable treatment periods showed a learning curve with a reduction of perforations in the second sequence (after 57 pedicle instrumentations) of about 15%, this was not found to be statistically significant. The fluoroscopic navigation of pedicle screws is a safe procedure at the lumbar spine with equal accuracy compared to the non-navigated conventional instrumentation. Application of C-arm navigation at the thoracic spine showed more inaccuracies, so that 3D-based navigation seems to be more advantageous in this region.

[Clinical accuracy of fluoroscopic navigation at the thoracic and lumbar spine]. / Klinische Präzision der C-Arm-Navigation an der Brust- und Lendenwirbelsäule.

The aim of the present study was the verification of the accuracy of 2-D fluoroscopy-based navigated pedicle screws at the thoracic and lumbar spine in a case series of traumatised patients. Within 36 months 111 pedicle screws in 29 patients were instrumented using C-arm based navigation, 60 at the thoracic and 51 at the lumbar spine. All screw positions were evaluated postoperatively by a routine thin slice CT scan using multiplanar reconstruction. The position of a screw in relation to its pedicle was classified in a) screw completely intraosseous, b) screw perforated less than thread level and c) screw perforated more than thread level. In 34 thoracic (56.7%) and 32 lumbar (62.7%) screws complete intraosseous placement was observed, 14 thoracic (23.3%) and 14 lumbar (27.5%) screws perforated less than thread level. Perforations more than thread level were found in 12 thoracic (20%) and 5 lumbar (9.8%) screws. Only medial and lateral perforations of the pedicle were documented (without neurological signs), cranial or caudal ones did not occur. Segmentation of the C-arm navigation into two comparable treatment periods showed a learning curve with a reduction of perforations in the second sequence (after 57 pedicle instrumentations) of about 15%, this was found to be not statistically significant. The fluoroscopic navigation of pedicle screws is a safe procedure at the lumbar spine with equal accuracy compared to the non-navigated conventional instrumentation. Application of C-arm navigation at the thoracic spine showed more inaccuracies, so that 3-D-based navigation seems to be advantageous in this region.

Biomechanical comparison of two side plate fixation techniques in an unstable intertrochanteric osteotomy model: Sliding Hip Screw and Percutaneous Compression Plate.

BACKGROUND: Our objective was to determine the biomechanical primary stability of two different side plate fixation devices in an unstable intertrochanteric cadaver model: the Sliding Hip Screw with an additional derotation screw was compared with the Percutaneous Compression Plate. METHODS: Eight pairs of human cadaver femurs were tested for comparison of the primary stability of the two implants in two modes: (a) cyclic loading up to 200 N, 400 N, 600 N, 800 N, and 1000 N, respectively. In vitro combined axial and bending loads were applied. Angular displacements of the proximal head fragment during loading were recorded in rotational, varus-valgus, and anterior-posterior directions. (b) The load to failure was determined. FINDINGS: Specimens fixed with the Percutaneous Compression Plate showed higher displacements in the varus during loading up to 200 N (P=0.033), and 400 N (P=0.001), compared to the Sliding Hip Screw. A similar tendency was observed for higher loads. The Percutaneous Compression Plate allowed more external rotation of the proximal fragment only at loads up to 800 N (P=0.019). No statistical difference could be found for the slight migrations in the posterior direction. Load to failure also revealed no statistical difference between the two implants. INTERPRETATION: The Percutaneous Compression Plate as a double-axis fixation device with a sliding capability allows higher displacements in the varus direction and also in external rotation at 800 N loading compared to the Sliding Hip Screw as a single-axis fixation device combined with an additional derotation screw. While both implants are successful used in clinical practice, this should be considered in treatment of unstable intertrochanteric fractures with inferior comminution in osteoporotic patients.

[Therapy indications and options for skeletal metastases]. / Therapieindikationen und -optionen bei ossären Metastasen.

Pathological fractures are dreaded complications due to skeletal metastasis. Modern oncological therapies and more sophisticated new radiological techniques such as MRI and positron emission tomography have improved multimodal treatment concepts. Surgical intervention is determined by the primary disease, general condition and life expectancy of the patient. The goals of surgical treatment are improvement of life quality, pain relief and maintenance of mobility.

Hip arthroplasty with proximal transmission of force: first clinical results with a new partially cemented femoral stem.

BACKGROUND: Proximal stem fixation by partial cementing is a new concept in hip arthroplasty. We conducted a prospective clinical and radiological analysis to evaluate the preliminary outcome of this new technique with the Option 3000 stem (Mathys Orthopaedics, Bettlach, Switzerland). METHODS AND FOCUS: One hundred and thirty-three hip replacements in 123 patients have been performed between 1996 and 2003: All of them were followed up regularly both clinically and radiological and 53 were analysed with the EBRA-FCA method. Eighty-six patients with 95 hips could be seen in August 2004. At this point of time, the mean follow-up time was 61 months (5.08 years) with a maximum of 100 months (8.33 years) RESULTS: The clinical data reported an average Harris Hip Score of 85.5. Nine stems had to be exchanged over the period of study. The EBRA-FCA analysis reported a mean subsidence less than 1.5 mm after 2 years, then an average stable subsidence of 2.4 mm. So the results are similar to the early results obtained with other fixation concepts and the long-term results appear promising.

[Computer-assisted arthrodesis of the sacroiliac joint]. / Navigationsgestützte Arthrodese des Iliosakralgelenks.

Arthrodesis of the sacroiliac joint (SI) usually requires a large surgical exposure using the lateral approach. Computer-assisted surgery based on intraoperative 3D fluoroscopy imaging can reduce the approach to stab incisions. The clinical example shows the insertion of two screws and a cylindrical bone graft to achieve an arthrodesis of the SI joint. The intraoperatively navigated placement of implants and bone graft was performed only by stab incisions.

Crystal structures of beta-galactosidase from Penicillium sp. and its complex with galactose.

Beta-galactosidases catalyze the hydrolysis of beta(1-3) and beta(1-4) galactosyl bonds in oligosaccharides as well as the inverse reaction of enzymatic condensation and transglycosylation. Here we report the crystallographic structures of Penicillium sp. beta-galactosidase and its complex with galactose solved by the SIRAS quick cryo-soaking technique at 1.90 A and 2.10 A resolution, respectively. The amino acid sequence of this 120 kDa protein was first assigned putatively on the basis of inspection of the experimental electron density maps and then determined by nucleotide sequence analysis. Primary structure alignments reveal that Penicillium sp. beta-galactosidase belongs to family 35 of glycosyl hydrolases (GHF-35). This model is the first 3D structure for a member of GHF-35. Five distinct domains which comprise the structure are assembled in a way previously unobserved for beta-galactosidases. Superposition of this complex with other beta-galactosidase complexes from several hydrolase families allowed the identification of residue Glu200 as the proton donor and residue Glu299 as the nucleophile involved in catalysis. Penicillium sp. beta-galactosidase is a glycoprotein containing seven N-linked oligosaccharide chains and is the only structure of a glycosylated beta-galactosidase described to date.

[Operative results of treatment of intraarticular calcaneus fractures with the "low contact plate"]. / Ergebnisse nach Osteosynthese intraartikulärer Kalkaneusfrakturen mit der "Low contact" Platte.

Calcaneus fractures are a major cause of accident- related invalidity. Long-term results are unsatisfactory with various therapy concepts as reported in the literature. Objective of this study is the long-term clinical and radiological outcome of a patient group treated with a low contact plate. Subjective assessment of function by the patient was an important evaluation criterion. From 01/92 to 12/98 a total of 182 patients with 209 calcaneus fractures were treated. Out of them 95 patients with 109 calcaneus fractures had a follow-up over a period ranging from 6 to 88 months (average 37.9 months). Severe impairment of walking was reported by 34.7 % of the patients. Severe impairment for work was declared by 68.4 % of the patients. 74.7 % of the patients complained middle or severe burden-pain. 64.3 % of the patients felt their quality of life impaired. After the injury 75.8 % of the patients were still employed. Arthrosis of the inferior ankle joint was found in 91.5 % of the cases. Within this collective three patient groups were formed with different follow-up periods to evaluate the temporal course, showing no significant differences. Frequently good functional and subjective results cannot be expected even with standardized operative procedures.

Difference in metallic wear distribution released from commercially pure titanium compared with stainless steel plates.

INTRODUCTION: Stainless steel and commercially pure titanium are widely used materials in orthopedic implants. However, it is still being controversially discussed whether there are significant differences in tissue reaction and metallic release, which should result in a recommendation for preferred use in clinical practice. MATERIALS AND METHODS: A comparative study was performed using 14 stainless steel and 8 commercially pure titanium plates retrieved after a 12-month implantation period. To avoid contamination of the tissue with the elements under investigation, surgical instruments made of zirconium dioxide were used. The tissue samples were analyzed histologically and by inductively coupled plasma atomic emission spectrometry (ICP-AES) for accumulation of the metals Fe, Cr, Mo, Ni, and Ti in the local tissues. Implant corrosion was determined by the use of scanning electron microscopy (SEM). RESULTS: With grades 2 or higher in 9 implants, steel plates revealed a higher extent of corrosion in the SEM compared with titanium, where only one implant showed corrosion grade 2. Metal uptake of all measured ions (Fe, Cr, Mo, Ni) was significantly increased after stainless steel implantation, whereas titanium revealed only high concentrations for Ti. For the two implant materials, a different distribution of the accumulated metals was found by histological examination. Whereas specimens after steel implantation revealed a diffuse siderosis of connective tissue cells, those after titanium exhibited occasionally a focal siderosis due to implantation-associated bleeding. Neither titanium- nor stainless steel-loaded tissues revealed any signs of foreign-body reaction. CONCLUSION: We conclude from the increased release of toxic, allergic, and potentially carcinogenic ions adjacent to stainless steel that commercially pure Ti should be treated as the preferred material for osteosyntheses if a removal of the implant is not intended. However, neither material provoked a foreign-body reaction in the local tissues, thus cpTi cannot be recommend as the 'golden standard' for osteosynthesis material in general.

[Precision of navigation-assisted surgery of the thoracic and lumbar spine]. / Präzision der navigationsgestützten Chirurgie an der Brust- und Lendenwirbelsäule.

The goal of these studies was to evaluate the accuracy of in vivo and in vitro application of CT- and C-arm-based navigation at the thoracic and lumbar spine. With CT based navigation, 82 pedicle screws were consecutively inserted, 53 into the thoracic and 29 into the lumbar spine. Seven (13%) perforations were detected at the thoracic spine and two (7%) at the lumbar spine. Additionally, minor perforations below the thread depth were seen in six (11%) thoracic and in two (7%) lumbar instrumentation. With C-arm-based navigation, 74 screws were consecutively placed into 38 thoracic and 36 lumbar pedicles. Perforations were noted in ten (26%) thoracic and four (11%) lumbar implants. Minor perforations were observed in another nine (24%) thoracic and ten (28%) lumbar pedicles. The observer-independent and standardized in vitro study based on a transpedicular 3.2-mm drill hole aiming a 4-mm steel ball in a plastic bone model showed pedicle perforations of the drill canal only in thoracic vertebrae, 1 of 15 in CT-based and 3 of 15 in C-arm navigation. The quantitative calculation of the smallest distance between the central line through the drill canal and the center of the steel ball resulted in 1.4 mm (0.5-4.8 mm) for the CT-based navigation at the thoracic spine and in 1.8 mm (0.5-3 mm) at the lumbar spine. For the C-arm based navigation the distance was 2.6 mm (0.9-4.8 mm) for the thoracic spine and 2 mm (1.2-3 mm) for the lumbar spine. In our opinion, the clinical results of the comparative accuracy of CT- and C-arm-based navigation in the present study showed moderate advantages of the CT-based technique in the thoracic spine, whereas CT- and C-arm based navigation had comparable perforation rates at the lumbar pedicle. The results of the experimental study correlated with the clinical data.

[Navigation of tumors and metastases in the area of the thoraco-lumbar spine]. / Navigation von Tumoren und Metastasen im Bereich der thorakolumbalen Wirbelsäule.

In this clinical feasibility study, CT-based verification of the efficacy of navigated decompression and pedicle screw placement in patients who had tumor-related posterior surgery was demonstrated. Eighty-six percent of the pedicle screws were positioned centrally in the bone without perforation; in all patients accurate decompression was seen. The accuracy of transpedicle screw implantation postoperatively was investigated with CT. In contrast to other published studies, no postoperative neurologic deterioration was seen in the patients as a result of using computer-aided surgical procedures. At the same time we were able to achieve complete decompression of the neural structures for radiologic and neurologic findings. Because of inaccurate registration, it was not possible to use computer-aided implantation surgery for 15% of the pedicles and, therefore, a conventional fluoroscopic approach was used. Our initial results indicate that computer-aided frameless navigation of tumor surgery of the spine is a safe technique which improves surgical performance during posterior decompression and transpedicle stabilization. In addition, CAS surgery improved the intraoperative information about the tumor and the current surgical intervention during decompression. Nevertheless the technique should be used only by experienced surgeons who can, if required, continue the operation using conventional techniques. Furthermore, the surgeon should have a complete theoretical understanding of the navigation system to minimize possible misinterpretation of computer guidance information.

[Radiation dosage in orthopedics -- a comparison of computer-assisted procedures]. / Strahlendosis im OP--ein Vergleich computerassistierter Verfahren.

Since the 1990s, computer-assisted methods have been well-established in the field of orthopedics and traumatology. In addition to the proven improvement in the field of surgical precisioncompared with conventional techniques, the decrease in the amount of emitted ionizing radiation inside of the operating room was cited to constitute an additional major advantage of computer-based navigation. The goal of this study was to quantify X-ray dose values exposed during both conventional methods and computer-assisted procedures, the latter being performed using a C-arm device SIREMOBIL Iso-C3D (Siemens Medical Solutions). The clinical measurements were supplemented by laboratory experiments using a pinpoint ionization chamber placed inside a standard lucite phantom. The clinical part of the study investigated the application of new, i.e. computer assisted, techniques in comparison with conventional procedures on in total 42 patients experiencing surgery either on their lower extremities (n = 14) or on the spine (n = 28). The techniques applied were: (1) conventional surgical procedure, (2) CT-based navigation, (3) C-arm navigation, and (4) Iso-C3D-navigation. The main focus of the clinical evaluation was the determination of radiation dose values by means of thermoluminescence dose meters, accompanied by the registration of the operation times of the X-ray tube during a surgical intervention. The conclusions from the clinical studies are in agreement with the results from the laboratory measurements. Computer-assisted techniques led to a significant reduction in X-ray dose exposure and drastically shortened irradiation times. The most pronounced decrease in the emission of ionizing radiation was achieved in case of the Iso-C3D-navigation, which has only recently been introduced into clinical practice, i.e. during runtime of this study.

Monitoring and healing analysis of 100 tibial shaft fractures.

BACKGROUND: We assessed the value of measuring biomechanical stiffness by assessing the fixator's external deformation as an objective means for monitoring fracture healing and determining the postoperative treatment regime, as compared to clinical and radiographic means of evaluation. PATIENTS AND METHODS: One hundred patients with tibial shaft fractures managed by unilateral external fixation had their fracture stiffness monitored. Stiffness was measured and clinical and radiological examinations were performed every 3-4 weeks. RESULTS: The time required for healing as indicated by stiffness measurement was an average of 2.5 weeks earlier than by radiological assessment. Eighty-two patients healed within 19 weeks (12.1+/-3.3 weeks) and ten patients in the following 6 weeks (24+/-4.3 weeks). Eight patients did not show an increase in fracture stiffness and received intramedullary nailing at a second operation. The average healing time was 11.3+/-4 weeks for type A, 13.1+/-3.6 weeks for type B fractures, and 15.1+/-5.9 weeks for type C fractures. The healing time for closed fractures was 11.3+/-3.2 weeks and for open fractures 14+/-4.9 weeks. CONCLUSIONS: The measurement of fracture stiffness allows the detection of patients at risk for nonunions. The healing time increased with increasing fracture gap size and was less in patients with younger age, less complex fractures, and lesser degrees of soft tissue damage.

[Combined posterior and anterior endoscopic supported instrumentation of the thoracic and lumbar spine. Arguments, concepts and first clinical results]. / Die dorsoventrale endoskopisch unterstützte Instrumentation der BWS und LWS. Argumente, Versorgungskonzepte und erste klinische Ergebnisse.

The operative therapeutic principles of thoracic and lumbar spine injuries are based on reposition, decompression and stabilization of the unstable area. Secondary loss of correction and consecutive deformation have negative impact on the long-term results after defect fractures of vertebral bodies and will be minimized only by the reconstruction of all involved spinal columns. With dissemination of thoracoscopic techniques at the thoracic spine and minimal invasive retroperitoneal approaches at the lumbar spine a decrease of the morbidity was achieved with equal effectivity for recalibration and fusion. Our experiences with the first consecutive 42 patients, treated minimal invasive are presented. Indications for anterior thoracoscopic and minimal invasive instrumentations after posterior transfixation are remaining osseous defects of the end plates of more than a quarter of the volume of the involved vertebra in case of migration of the vertebral disc, wedging of the vertebral body after posterior reposition of more than 10 degrees and persisting anterior encroachment of the spinal canal of more than 30 %. The decision is based on radiographs and CT-scans, performed after posterior stabilization.

[Sources of error and risks in CT based navigation]. / Fehlerquellen und Gefahren der CT-basierten Navigation.

Based on the experience of 4 cervical, 102 thoracic/lumbar pedicle screw and 14 transiliosacral screw implantations all problems and complications were collected. Problems noted within the data collection in the preoperative CT were an incomplete acquisition of the surgical target (n = 3), an exceeding of the processable scan slices (n = 1) and a non focused field of view. Transmission of the CT datas often were documented as incomplete (n = 16). Segmentation of the CT dataset turned out to be the significant problem with incorrect differentiation of the bone-soft tissue transition (n = 2), where as the choice of the matching points and the trajectories did not provoke any mistakes in the planning modus. The intraoperative matching of both corresponding datasets was insufficient (n = 7), while the assignment of the CT dataset to the correct vertebral was not a major problem (n = 1). Navigation was not possible (n = 2) due to an instability of the spinal process. All fiducial based matching procedures (pelvis) were carried out without any problems. During intraoperative navigation potential complications resulted from deformation of instruments (n = 1) and interaction of instruments and the data reference base (n = 2). Further, the CT-based navigation of fractured vertebrae or unstable iliosacral joints is not safe, because dislocations between acquisition of the dataset and operation will lead to misguidance.

Spinal navigation in cervical fractures--a preliminary clinical study on Judet-osteosynthesis of the axis.

OBJECTIVE: To evaluate the accuracy of CT-based and computer-aided screw insertion into the pedicles of the axis using the method of Judet. Materials and Methods In two patients, four transpedicular implants of C2 were positioned using a computer-guided technique. One patient with iatrogenic destabilization of the posterior structures C3 and C4 and one patient with a hangman's fracture of the axis required pedicular fixation. In addition, intraoperative documentation of the additional time required for the navigation procedures was made. Finally, postoperative CTs of each patient provided further information about transpedicular implant localization. RESULTS: Image-guided implantation of screws was possible in all scheduled pedicles of the axis. In the postoperative CT scans, none of the inserted screws perforated the medial or lateral pedicle. All screws were accurately positioned within the pedicles, and no anterior perforation of the screws into the vertebral foramen was observed. CONCLUSIONS: Our initial results using computer-aided implantation of Judet screws showed that these screws were inserted correctly. However, it is important to realize that movements can occur in the vertebral arch and lateral mass during the time between preoperative CT and surgery. This possibility for movement should be taken into account when procedures such as CT-based insertion of image-guided screws into patients with fractures of the isthmus C2 are performed, because damage to neurovascular structures can result. Therefore, this navigation system should only be used by experienced surgeons who can, if necessary, continue the intervention with more conventional techniques. In addition, it is essential that the surgeon have a complete understanding of the principles of the tracking systems to prevent possible misinterpretation of computer-generated information.

Quantitative assessment of the expression of melanoma-associated antigens by non-competitive reverse transcription polymerase chain reaction.

The assessment of tumor-associated antigens (TAA) recognized by T lymphocytes is a prerequisite for diagnosis and immunotherapy of melanoma. Different reverse transcription-polymerase chain reaction (RT-PCR) protocols allowing the quantification of the TAA mRNA expression in the solid tumor or the detection of circulating melanoma cells have been described. We have recently shown a positive correlation between the amount of specific product formed by RT-PCR and the staining intensity in immunohistochemical analysis of the corresponding sample. Here we describe a quantification procedure based on the direct digitization of the PCR products after separation on ethidium bromide-stained agarose gels, followed by computer-assisted densitometry. To standardize our method, we examined the linear range of the densitometric quantification procedure as reflected by the correlation of signal intensity to the amount of the corresponding DNA. As an internal measure for the so-termed cDNA in the different samples after RNA isolation and reverse transcription, a beta-actin PCR was introduced. Subsequently, we chose four sets of primers for the melanoma-associated antigens MAGE1, tyrosinase, Melan A/MART-1 and gp100/Pmel17 and performed PCR analysis over a range of cycle numbers. In each case, the amplification rate remained constant up to at least 26 cycles under the respective conditions. Plotting the logarithm of the amount of product against the cycle number yields a slope that equals the logarithm of the amplification rate. The amount of starting material can be determined from the intercept with the ordinate. In summary, the method introduced in the present work allows the quantification of TAA in melanoma which might be important for the monitoring of disease. Technically the method is sound and sensitive, avoids post-PCR manipulations and can be performed with the standard equipment of a molecular biology laboratory. It can be applied also to other solid tumors and leukemias.