Technical report: surgical preparation of human brain tissue for clinical and basic research.

BACKGROUND: The study of the distinct structure and function of the human central nervous system, both in healthy and diseased states, is becoming increasingly significant in the field of neuroscience. Typically, cortical and subcortical tissue is discarded during surgeries for tumors and epilepsy. Yet, there is a strong encouragement to utilize this tissue for clinical and basic research in humans. Here, we describe the technical aspects of the microdissection and immediate handling of viable human cortical access tissue for basic and clinical research, highlighting the measures needed to be taken in the operating room to ensure standardized procedures and optimal experimental results. METHODS: In multiple rounds of experiments (n = 36), we developed and refined surgical principles for the removal of cortical access tissue. The specimens were immediately immersed in cold carbogenated N-methyl-D-glucamine-based artificial cerebrospinal fluid for electrophysiology and electron microscopy experiments or specialized hibernation medium for organotypic slice cultures. RESULTS: The surgical principles of brain tissue microdissection were (1) rapid preparation (<1 min), (2) maintenance of the cortical axis, (3) minimization of mechanical trauma to sample, (4) use of pointed scalpel blade, (5) avoidance of cauterization and blunt preparation, (6) constant irrigation, and (7) retrieval of the sample without the use of forceps or suction. After a single round of introduction to these principles, multiple surgeons adopted the technique for samples with a minimal dimension of 5 mm spanning all cortical layers and subcortical white matter. Small samples (5-7 mm) were ideal for acute slice preparation and electrophysiology. No adverse events from sample resection were observed. CONCLUSION: The microdissection technique of human cortical access tissue is safe and easily adoptable into the routine of neurosurgical procedures. The standardized and reliable surgical extraction of human brain tissue lays the foundation for human-to-human translational research on human brain tissue.

NF-kB decoy oligodeoxynucleotide preserves disc height in a rabbit anular-puncture model and reduces pain induction in a rat xenograft-radiculopathy model.

While it is known that the degenerated intervertebral disc (IVD) is one of the primary reasons for low-back pain and subsequent need for medical care, there are currently no established effective methods for direct treatment. Nuclear factor-κB (NF-κB) is a transcription factor that regulates various genes' expression, among which are inflammatory cytokines, in many tissues including the IVD. NF-κB decoy is an oligodeoxynucleotide containing the NF-κB binding site that entraps NF-κB subunits, resulting in suppression of NF-κB activity. In the present preclinical study, NF-κB decoy was injected into degenerated IVDs using the rabbit anular-puncture model. In terms of distribution, NF-κB decoy persisted in the IVDs up to at least 4 weeks after injection. The remaining amount of NF-κB decoy indicated that it fit a double-exponential-decay equation. Investigation of puncture-caused degeneration of IVDs showed that NF-κB decoy injection recovered, dose-dependently, the reduced disc height that was associated with reparative cell cloning and morphological changes, as assessed through histology. Gene expression, by quantitative real-time polymerase chain reaction (qRT-PCR), showed that NF-κB decoy attenuated inflammatory gene expression, such as that of interleukin-1 and tumor necrosis factor-α, in rabbit degenerated IVDs. NF-κB decoy also reduced the pain response as seen using the "pain sensor" nude rat xenograft-radiculopathy model. This is the first report demonstrating that NF-κB decoy suppresses the inflammatory response in degenerated IVDs and restores IVD disc height loss. Therefore, the intradiscal injection of NF-κB decoy may have the potential as an effective therapeutic strategy for discogenic pain associated with degenerated IVDs.

[Diagnosis and treatment of periprosthetic infections]. / Diagnose und Therapie periprothetischer Infektionen.

The differentiation of periprosthetic infections into early onset and late onset infections is the determining factor fores the therapeutic strategy. The diagnosis of chronic unapparent infections is sometimes difficult. The therapeutic success, especially in early onset infections, is essentially influenced by a well-timed diagnosis and an immediate therapy. Surgical wound debridement with preservation of the prosthesis, one-stage replacement and two-stage replacement all with concomitant appropriate systemically administered double combined antibiotics are therapeutic options depending on factors, such as the duration of the infection. Prosthesis replacement is currently the only option to treat biofilm colonization of endoprosthetic surfaces. The bacterial adhesion on polyethylene can be extremely resistant. Due to the therapeutic consequences, periprosthetic infections remain one of the worst complications in endoprosthetic joint replacement.

[Midcarpal fusion with the spider plate]. / Mediokarpale Teilarthrodese des Handgelenks mittels Spiderplatte.

OBJECTIVE: Pain relief while preserving wrist motion in advanced carpal collapse. INDICATIONS: Advanced carpal collapse stage II/III due to scaphoid non-union, scapholunate ligament tear, idiopathic radiocarpal osteoarthritis, aseptic osteonecrosis of the scaphoid (Preisser's disease). A relative indication is chronic midcarpal instability. CONTRAINDICATIONS: Osteoarthrisis of the lunate. Radiocarpal instability with ulnar translation of the wrist. SURGICAL TECHNIQUE: Dorsal curved incision between the 3rd and 4th dorsal extensor compartment. Partial wrist denervation (posterior interosseous nerve). Raising of a radial pedicled capsule flap. Complete extirpation of the scaphoid without fragmentation. Cartilage removal of all the joint facets for arthrodesis. Reduction of the lunate and temporary fixation with K wires between the triquetrum and capitate and the triquetrum and lunate. Milling the plate hole exactly in central position of the four carpal bones. The plate should not protrude from the bone to avoid impingement with the dorsal limb of the radius. Transferring of spongiosa harvested from the removed scaphoid into the arthrodesis gap. Fixation of the 8-hole plate with 2 screws in each of the four carpal bones. Fluoroscopy of the screw fitting. Control of the correct wrist articulation (motion test). Lavage of the wrist. Suture of the capsule flap. Redon drain, wound closure, dorsal splinting. POSTOPERATIVE MANAGEMENT: Dorsal splint for 3 weeks; finger mobilization up to complete fist closure starting on postoperative day 1. From week 4-6, active wrist motion, from week 7-10 with increasing load. Return to work after 11-12 weeks. CT scan in cases of delayed union, abnormal function, or persistence of pain. RESULTS: Between 2002 and 2008, 36 four-corner fusions (32 male, 4 female) with the spider plate were performed in 24 right and 12 left wrists. The mean age of the patients at surgery was 48 years (range 32-71 years). Follow-up examinations were performed in 11 patients. The Krimmer and Rudolf scores were determined: 1 excellent, 7 good, 2 satisfactory, and 1 poor result were found. The mean grip strength was 51% relative to the opposite wrist. The mean range of motion (ROM) for wrist extension/flexion was 56% of the opposite side. Wrist extension/flexion averaged 24/0/32° and ulnar/radial deviation was on average 20/0/19°. Pain during activity was on average 2.2 based on the visual analogue scale (0-10). One delayed bony union due to a screw breakage, which required total wrist fusion, was observed.

Are psychoeducational smoking cessation interventions for coronary heart disease patients effective? Meta-analysis of interventions.

PURPOSE: This systematic review aimed to assess the effectiveness of psychoeducational smoking cessation interventions for coronary heart disease (CHD) patients; and to examine behaviour change techniques used in interventions and their suitability to change behavioural determinants. METHODS: Multiple bibliographic databases and references of retrieved articles were searched for relevant randomized controlled studies. One reviewer extracted and a second reviewer checked data from included trials. Random effects meta-analyses were conducted to estimate pooled relative risks for smoking cessation and mortality outcomes. Behaviour change techniques used and their suitability to change behavioural determinants were evaluated using a framework by Michie, Johnston, Francis, Hardeman, and Eccles. RESULTS: A total of 14 studies were included. Psychoeducational interventions statistically significantly increased point prevalent (RR 1.44, 95% CI, 1.20-1.73) and continuous (RR 1.51, 95% CI, 1.18-1.93) smoking cessation, and statistically non-significantly decreased total mortality (RR 0.73, 95% CI, 0.46-1.15). Included studies used a mixture of theories in intervention planning. Despite superficial differences, interventions appear to deploy similar behaviour change techniques, targeted mainly at motivation and goals, beliefs about capacity, knowledge, and skills. CONCLUSIONS: Psychoeducational smoking cessation interventions appear effective for patients with CHD. Although questions remain about what characteristics distinguish an effective intervention, analysis indicates similarities between the behaviour change techniques used in such interventions.

Improving the efficacy of cancer immunotherapy.

A series of cancer vaccines have been evaluated in clinical trials with encouraging results, but the demonstration of clinical benefit in confirmatory studies has so far proven to be difficult. The development of cancer vaccines is hampered by a range of issues particular to this field of research. On 12th March 2008, the Biotherapy Development Association convened a workshop to discuss issues faced by scientists and clinicians involved in the development of cancer vaccines. This paper is a review of the field, based on discussions held at the BDA workshop, and describes biological barriers encountered in generating effective immune responses to tumours, methodological obstacles encountered in the improvement of immunological monitoring which aims to improve inter-laboratory and inter-trial comparisons, challenges in clinical trial design and problems posed by the lack of specific regulation for cancer vaccines and the impact on their development. Ultimately, a number of general solutions are posed: (1) better patient selection, (2) use of multi-modal treatments that affect several aspects of the immune system at once, (3) a requirement for the development of good biomarkers to stratify patients for selection prior to trial and as surrogates for clinical response and (4) harmonisation of SOPs for immunological monitoring of clinical trials.

Hypericin activated by an incoherent light source has photodynamic effects on esophageal cancer cells.

BACKGROUND AND AIMS: Photodynamic therapy (PDT) is a new treatment modality for early esophageal neoplasia. With two absorption maxima in the visible light range (550 and 588 nm) hypericin is a very promising photosensitizer for PDT with incoherent light sources. We studied the effects of photosensitizing hypericin in both primary cell cultures and cell lines (squamous: Kyse-140 and adenocarcinoma: OE-33) of human esophageal cancer using an incoherent white light source. MATERIALS AND METHODS: Esophageal cancer cells were preincubated (4-24 h) with hypericin (10 nM-1 micro M) and then irradiated with a light energy dose of 30 J/cm(2). RESULTS: Hypericin showed strong phototoxic effects and induced apoptosis in a dose-dependent fashion. The IC(50) value of hypericin phototoxicity was approximately 30 nM in both squamous and adenocarcinoma cells. In the concentrations used nonphotoactivated hypericin showed no toxic or apoptotic potency. The phototoxicity of hypericin was compared to that of delta-aminolevulinic acid (5-ALA), which is already being used for photodynamic therapy of gastrointestinal cancer. 5-ALA produced similar phototoxic effects but at a much higher dose (IC(50) 182+/-8 micro M in Kyse-140 and 308+/-40 micro M in OE-33 cells). Moreover, 5-ALA did not induce apoptosis to a relevant extent. CONCLUSION: Hypericin is a very promising new photosensitizer for innovative photodynamic therapy of esophageal cancer. Both the well known clinical safety of hypericin and the lower costs of broad band light sources argue in favor of clinical trials.

Using Australian DRGs in Germany: a commentary.

Germany will begin a change to per case payment by DRG from January 2003. It has selected the Australian DRG classification as the basis for patient categorisation, in preference to the many other DRG variants around the world. The main aim is increase control over expenditure. We describe some of the reasons for high levels of spending on hospital inpatient care, including the fragmented insurance system and supplier-induced demand. We summarise the reasons why Australian DRGs were selected, and note some of the benefits that will accrue for Australia.

Pathogenesis and symptomatology of cholelithiasis in childhood. A prospective study.

QUESTIONS: How may gallstones develop if there is no hemolytic disorder and no malformation or alteration of the gallbladder and cystic ducts? Was there a change in the incidence of stones and the assignment of the patients to different age groups? The literature gives answers to these questions on the basis of only few cases. Therefore, a relatively large prospective database of collected cases during 30 years shall be used to give additional answers. METHODS: The data are derived from 111 children with symptomatic gallstones without hemolytic diseases. 109 of them required operative therapy. They were examined preoperatively and postoperatively maximum during 30 years of follow-up as well according to a defined program. This program includes taking the patient's history, evaluation of clinical, radiological including sonographical examination, exclusion of hemolytic primary diseases, histological investigation of intraoperatively gained specimens of the gallbladder, and the spectroscopic qualitative as well as semiquantitative analysis of stones and bile. RESULTS: (1) The qualitative and quantitative composition of the stones is different in the four age groups. (2) The causes of lithogenesis are different in the four age groups. (3) Children with long-term TPN, after cardiopulmonary bypass operations, after extended small bowel resection, suffering from obesity, and girls using hormonal contraception are at risk of developing gallstones. We recommend to observe these patients by repeated ultrasound controls for gallstone formation during a minimum of 10 years. (4) Malformations and pathological alterations of the gallbladder are rare causes for increased lithogenity. The gallbladder appeared morphologically normal in 61% of patients with symptomatic gallstones. (5) An increased frequency of gallstones during the last 30 years was only observed in the age group under 1 year.

[Diagnosis of glomus jugulare tumor recurrence with dynamic contrast medium flow in MRI]. / Rezidivdiagnostik von Glomus jugulare-Tumoren mittels dynamischer Kontrastmittel-Anflutung in der MRT.

PURPOSE: To evaluate the ability of dynamic MRI to differentiate residual tumor from scar after glomus jugulare (GJ) surgery. METHODS: 26 patients after GJ surgery were examined with dynamic contrast medium MRI on a 1.5 T Philips-Gyro-Scan NT. A T1-weighted TFE sequence was performed to localise the 5 slices for the dynamic study. Subsequently, a T1-weighted TFE sequence was used for the dynamic contrast medium study. We started contrast medium administration and TFE sequence at the same time. The time until acquisition of the first image was 7.4 s, and the interval between two images 7 s (total time 2.24 min). Intensity versus time studies were performed in the region of interest. RESULTS: 2 typical intensity versus time graphs were observed: residual tumors showed a rapid increase in signal intensity, followed by a "wash-out" phase, and a plateau phase in the later course. Scar showed a moderate increase in signal intensity, followed by a plateau phase on a low intensity level. In 9 patients, in whom tumor removal was subtotal, we saw the typical graph of a GJ tumor. 4 of 17 patients with complete tumor removal showed a typical graph like that of residual tumor, 13 of 17 patients showed the typical graph of a scar. CONCLUSION: With dynamic contrast medium MRI studies, differentiation between residual tumor and scar is possible in patients with prior GJ surgery.

Imaging of head and neck tumors--methods: CT, spiral-CT, multislice-spiral-CT.

Spiral-CT is standard for imaging neck tumors. In correspondence with other groups we routinely use spiral-CT with thin slices (3 mm), a pitch of 1.3-1.5 and an overlapping reconstruction increment (2-3 mm). In patients with dental fillings a short additional spiral parallel to the corpus of the mandible reduces artifacts behind the dental arches and improves the diagnostic value of CT. For the assessment of the base of the skull, the orbital floor, the palate and paranasal sinuses an additional examination in the coronal plane is helpful. Secondary coronal reconstructions of axial scans are helpful in the evaluation of the crossing of the midline by small tumors of the tongue base or palate. For an optimal vascular or tissue contrast a sufficient volume of contrast medium and a start delay greater than 70-80 s are necessary. In our opinion the best results can be achieved with a volume of 150 ml, a flow of 2.5 ml/s and a start delay of 80 s. Dynamic enhanced CT is only necessary in some special cases. There is clear indication for dynamic enhanced CT where a glomus tumor is suspected. Additional functional CT imaging during i-phonation and/or Valsalva's maneuver are of great importance to prove vocal cords mobility. Therefore, imaging during i-phonation is an elemental part of every thorough examination of the hypopharynx and larynx region. Multislice-spiral-CT allows almost isotropic imaging of the head and neck region and improves the assessment of tumor spread and lymph node metastases in arbitrary oblique planes. Thin structures (the base of the skull, the orbital floor, the hard palate) as well as the floor of the mouth can be evaluated sufficiently with multiplanar reformations. Usually, additional coronal scanning is not necessary with multislice-spiral-CT. Multislice-spiral-CT is especially advantageous in defining the critical relationships of tumor and lymph node metastases and for functional imaging of the hypopharynx and larynx not only in the transverse plane but also in the coronal plane.

2D and 3D visualisation of head and neck tumours from spiral-CT data.

PURPOSE: This paper intends to demonstrate the possibilities of two- and three-dimensional visualisation methods from spiral-CT data sets in the head and neck region and demonstrates their value based on various studies. MATERIAL AND METHODS: The scanner was a Somatom PLUS 4 (Siemens, Forchheim/Germany). The patients were scanned using a slice thickness of 3 mm and a table feed of 5 mm (pitch 1,6). The images were reconstructed with an increment of 1 mm. Contrast agent (150 ml) was applied intravenously with a flow of 2.5 ml/s and a start delay of 80 s. In one study the start delay was 20 s in order to visualise the carotid arteries (extracranial aneurysm). Volumetric data sets were postprocessed with 'Vitrea' and 'VoxelView' (Vital Images) volume rendering software on a Silicon Graphics O2 workstation (virtual laryngoscopy). Multiplanar reformation and colour-coded 3D-reconstruction were done on a Prominence workstation (Siemens/Forchheim/Germany). RESULTS: In every region of the head and neck MPRs are useful as additional planes (with the exception of the hypopharynx and the larynx), SSDs are always useful if there is extensive bony destruction (skull, spine, skeleton larynx). Colour-coded three-dimensional reformations may be done for extensive tumours and before multi-specialty surgery. Perspective volume rendering is already in use for virtual endoscopy of the paranasal sinuses and the virtual laryngoscopy. In temporal bone evaluation, perspective volume rendering is a new and promising method of the future to assess the inner ear. SUMMARY: Two- and three-dimensional displays are used to visualise pathological findings in their topographic relation to anatomical leading structures. Thus, the radiologist can point out to the clinician the pathological findings by some essential images without having to demonstrate all axial slices.

Methods: MRT.

MRI has become the imaging method of choice in special regions of the head and neck (e.g. nasopharynx, oropharynx, oral cavity, floor of the mouth). Superconducting MR-equipment with field strengths of 1.0-1.5 T are appropriate for the evaluation of the head and neck region. Signal acquisition is optimal with circular polarized head coils or with specially designed surface coils; the body coil is insufficient.When imaging tumors we need T1 contrast, T2 contrast and contrast medium information (enhancement information). For the T1 contrast T1-spin-echo is and remains the best sequence. For T2-contast T2 turbo-spin-echo with fat suppression has replaced the T2 spin-echo sequences because it is faster and shows good contrast between tumor and saturated fat tissue. Fat saturated T1 turbo-spin-echo enables best tissue contrast after Gd-DTPA application.

Paranasal sinuses and nasopharynx CT and MRI.

Neoplastic disease of the nose, paranasal sinuses, the nasopharynx and the parapharyngeal space requires thorough assessment of location and extent in order to plan appropriate treatment. CT allows the deep soft tissue planes to be evaluated and provides a complement to the physical examination. It is especially helpful in regions involving thin bony structures (paranasal sinuses, orbita); here CT performs better than MRI. MRI possesses many advantages over other imaging modalities caused by its excellent tissue contrast. In evaluating regions involving predominantly soft tissue structures (ec nasopharynx and parapharyngeal space) MRI is superior to CT. The possibility to obtain strictly consecutive volume data sets with spiral CT or 3D MRI offer excellent perspectives to visualize the data via 2D or 3D postprocessing. Because head and neck tumors reside in a complex area, having a 3D model of the anatomical features may assist in the delineation of pathology. Data sets may be transferred directly into computer systems and thus be used in computer assisted surgery.

Oropharynx, oral cavity, floor of the mouth: CT and MRI.

Pretherapeutic staging of tumors of the oropharynx, the oral cavity and the floor of the mouth is important and should be thorough and exact to ensure appropriate therapy. Particularly important is the assessment of infiltration of deeper compartments and the topographic relationship of tumor to vascular structures (lingual artery and vein, hypoglossal nerve), or the presence of spread of the tumor across the midline. As spread of tumor may occur to a large degree underneath normal appearing mucosa, clinical assessment of the true tumor extent is difficult. In the last 20 years computed tomography (CT) has proved its value as a supplementary non-invasive method and established its role in modern diagnostic evaluation. Magnetic resonance imaging (MRI) is an non-invasive scanning method that offers excellent tissue contrast. Ultrasonography (US) is of secondary importance, but provides useful guidance due to its wide availability and its easy use. This paper aims to depict the possibilities of modern CT and MRI to provide 'one-stop-shopping' information to the clinician as a basis for the right therapeutic approach and correct estimation of the individual patient's prognosis. A clear problem oriented imaging strategy with standardized diagnostic criteria will lead to a cost effective evaluation.

Head and neck tumors: imaging recurrent tumor and post-therapeutic changes with CT and MRI.

OBJECTIVE: To evaluate criteria for detection of tumor recurrence and post-treatment changes in patients with head and neck malignancies in computed tomography (CT) and magnetic resonance imaging (MRI). METHODS AND MATERIALS: Thirty-nine patients with head and neck carcinoma receiving radiochemotherapy were examined before, during and after therapy with MRI. Changes in signal intensity were correlated to histology or clinical course. Three hundred and thirty-one patients with head and neck malignancies were examined with CT after therapy. CT diagnoses were correlated with histology or clinical course. RESULTS: Main criteria for recurrent/residual tumor in MRI was infiltrative mass with high signal intensity in T2-weighted images and enhancement after Gd-DTPA in T1-weighted images. Radiation-induced changes led to false positive diagnosis in 46% in the interval up to 3 months after therapy and in 58% in the interval 3-6 months after therapy. The combination of a circumscribed, infiltrative mass with contrast enhancement in CT had a sensitivity of 86% and a specificity of 80%. CONCLUSION: CT could accurately demonstrate postoperative changes and tumor recurrence. MRI had advantages in differentiation of tumor and scar, but edema after radiation therapy can spoil diagnosis.

Synovial fluid levels of tumor necrosis factor alpha and oncostatin M correlate with levels of markers of the degradation of crosslinked collagen and cartilage aggrecan in rheumatoid arthritis but not in osteoarthritis.

OBJECTIVE: To compare synovial fluid (SF) levels of oncostatin M (OSM), tumor necrosis factor alpha (TNFalpha), and interleukin-6 (IL-6) in patients with rheumatoid arthritis (RA) and osteoarthritis (OA) and to determine which correlate best with SF levels of antigenic keratan sulfate (Ag KS), a marker of aggrecan catabolism, and pyridinium crosslinks, markers of the degradation of mature collagen molecules. METHODS: SF was drawn from the knee joints of patients with RA (n = 31) or OA (n = 31). Levels of Ag KS, D-pyridinoline (D-Pyr), pyridinoline (Pyr), OSM, TNFalpha, and IL-6 were measured by enzyme-linked immunosorbent assay. RESULTS: RA patients had higher median SF levels of OSM, TNFalpha, IL-6, and Pyr, but a lower median level of D-Pyr, than OA patients. In both groups, IL-6 levels correlated positively with those of OSM and TNFalpha. However, the correlation between levels of OSM and TNFalpha was only significant in the RA group. Ag KS and Pyr levels correlated positively in RA but not in OA. The correlation between TNFalpha and Ag KS was positive in RA and negative in OA. Further, in RA, OSM and IL-6 levels correlated strongly with Pyr and Ag KS levels but not with D-Pyr levels, while there were no strong correlations in OA for OSM or IL-6 levels with Pyr, Ag Ks, or D-Pyr levels. CONCLUSION: This in vivo study suggests that TNFalpha and other proinflammatory cytokines are involved in the up-regulation of the coordinated degradation of cartilage aggrecan and collagen in RA. Further, OSM may act synergistically with other proinflammatory cytokines in up-regulating the production of metalloproteinases by chondrocytes in rheumatoid joints.