Development of a preclinical orthotopic xenograft model of ewing sarcoma and other human malignant bone disease using advanced in vivo imaging.

Ewing sarcoma and osteosarcoma represent the two most common primary bone tumours in childhood and adolescence, with bone metastases being the most adverse prognostic factor. In prostate cancer, osseous metastasis poses a major clinical challenge. We developed a preclinical orthotopic model of Ewing sarcoma, reflecting the biology of the tumour-bone interactions in human disease and allowing in vivo monitoring of disease progression, and compared this with models of osteosarcoma and prostate carcinoma. Human tumour cell lines were transplanted into non-obese diabetic/severe combined immunodeficient (NSG) and Rag2(-/-/)γc(-/-) mice by intrafemoral injection. For Ewing sarcoma, minimal cell numbers (1000-5000) injected in small volumes were able to induce orthotopic tumour growth. Tumour progression was studied using positron emission tomography, computed tomography, magnetic resonance imaging and bioluminescent imaging. Tumours and their interactions with bones were examined by histology. Each tumour induced bone destruction and outgrowth of extramedullary tumour masses, together with characteristic changes in bone that were well visualised by computed tomography, which correlated with post-mortem histology. Ewing sarcoma and, to a lesser extent, osteosarcoma cells induced prominent reactive new bone formation. Osteosarcoma cells produced osteoid and mineralised "malignant" bone within the tumour mass itself. Injection of prostate carcinoma cells led to osteoclast-driven osteolytic lesions. Bioluminescent imaging of Ewing sarcoma xenografts allowed easy and rapid monitoring of tumour growth and detection of tumour dissemination to lungs, liver and bone. Magnetic resonance imaging proved useful for monitoring soft tissue tumour growth and volume. Positron emission tomography proved to be of limited use in this model. Overall, we have developed an orthotopic in vivo model for Ewing sarcoma and other primary and secondary human bone malignancies, which resemble the human disease. We have shown the utility of small animal bioimaging for tracking disease progression, making this model a useful assay for preclinical drug testing.

Fluid levels in medical imaging.

Fluid levels are commonly observed on a range of imaging methods in both normal and abnormal circumstances. Radiologists must be familiar with the appearances and significance of fluid levels, but more fundamentally, require an understanding of the mechanisms by which fluid levels occur and the principles necessary for the demonstration of fluid levels. These are the prerequisites of a cavity and at least two types of immiscible fluid, and most importantly, the requirement of an image that is orientated in the vertical plane.

A rare cause of spinal cord compression: imaging appearances of gout of the cervical spine.

Gout is a metabolic disorder typically affecting the peripheral joints, more commonly in males. Spinal involvement is uncommon and is usually associated with hyperuricemia. We present the imaging findings of a case of spinal gout in a female patient with no previous history of hyperuricaemia, involving multiple spinal segments.

Juvenile spondylolysis: a comparative analysis of CT, SPECT and MRI.

OBJECTIVE: To evaluate whether MRI correlates with CT and SPECT imaging for the diagnosis of juvenile spondylolysis, and to determine whether MRI can be used as an exclusive image modality. DESIGN AND PATIENTS: Juveniles and young adults with a history of extension low back pain were evaluated by MRI, CT and SPECT imaging. All images were reviewed blindly. Correlative analyses included CT vs MRI for morphological grading and SPECT vs MRI for functional grading. Finally, an overall grading system compared MRI vs CT and SPECT combined. Statistical analysis was performed using the kappa statistic. RESULTS: Seventy-two patients (mean age 16 years) were recruited. Forty pars defects were identified in 22 patients (31%), of which 25 were chronic non-union, five acute complete defects and ten acute incomplete fractures. Kappa scores demonstrated a high level of agreement for all comparative analyses. MRI vs SPECT (kappa: 0.794), MRI vs CT (kappa: 0.829) and MRI vs CT/SPECT (kappa: 0.786). The main causes of discrepancy were between MRI and SPECT for the diagnosis of stress reaction in the absence of overt fracture, and distinguishing incomplete fractures from intact pars or complete defects. CONCLUSIONS: MRI can be used as an effective and reliable first-line image modality for diagnosis of juvenile spondylolysis. However, localised CT is recommended as a supplementary examination in selected cases as a baseline for assessment of healing and for evaluation of indeterminate cases.

Percutaneous vertebroplasty: history, technique and current perspectives.

Percutaneous vertebroplasty is a safe and efficacious technique for the treatment of persistent pain from a fractured vertebral body. Injection of cement into the vertebral body is made after insertion of a large-bore needle, frequently by a trans-pedicular approach. Vertebroplasty is most commonly used to treat painful osteoporotic fracture resistant to conservative therapy, but may be helpful in other conditions such as malignant collapse. NICE guidelines are now available for this procedure, which is relatively new in the UK, but has been performed for more than 15 years in continental Europe.

Nitric oxide synergistically potentiates interleukin-1 beta-induced increase of cyclooxygenase-2 mRNA levels, resulting in the facilitation of substance P release from primary afferent neurons: involvement of cGMP-independent mechanisms.

We previously demonstrated that cultured rat dorsal root ganglion (DRG) cells respond to stimulation with interleukin-1 beta (IL-1 beta) by releasing substance P (SP), and this response is regulated via the cyclooxygenase (COX)-2 pathway. In this study, to ascertain the interaction between nitric oxide (NO) and prostaglandins in primary afferent neurons, we investigated the effect of NO on the IL-1 beta-induced release of SP in cultured DRG cells. An NO donor, S-nitroso-N-acetyl-DL-penicillamine (SNAP), did not in itself evoke SP release. However, it potentiated the IL-1 beta-induced release of SP. Similarly, while SNAP did not elicit the expression of COX-2 mRNA, it potentiated the expression induced by IL-1 beta in cultured DRG cells, and this potentiation was significantly suppressed by the NO scavenger, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (carboxy-PTIO). Moreover, SNAP also potentiated the expression of COX-2 protein induced by IL-1 beta in cultured DRG cells. The stimulatory effect of SNAP on the IL-1 beta-induced release of SP was completely inhibited on co-incubation with a selective COX-2 inhibitor, NS-398. 1H-[1,2,4]oxadiazolo[4,3-a]quinoxaline-1-one (ODQ), a potent inhibitor of soluble guanylate cyclase, did not suppress, and a membrane-permeable cGMP analogue, 8-Br-cGMP, did not mimic the stimulatory effects of SNAP in DRG cells. These results suggest that in cultured DRG cells, NO potentiates the IL-1 beta-induced increase in COX-2 expression via a soluble guanylate cyclase-cGMP-independent pathway, resulting in facilitation of SP release. The interaction between NO and COX in primary afferent neurons might contribute to the change in nociceptive perception in inflammatory hyperalgesia.

The role of diagnostic radiology in compressive and entrapment neuropathies.

Diagnostic imaging is increasingly being utilised to aid the diagnosis of compression and entrapment neuropathies. Cross-sectional imaging, primarily ultrasound and magnetic resonance imaging, can provide exquisite anatomical detail of peripheral nerves and the changes that may occur as a result of compression. Imaging can provide a useful diagnostic aid to clinicians, which may supplement clinical evaluation, and may eventually provide an alternative to other diagnostic techniques such as nerve conduction studies. This article describes the abnormalities that may be demonstrated by current imaging techniques, and critically analyses the impact of imaging in diagnosis of peripheral compressive neuropathy.

Fully automated software to monitor wear in prosthetic knees using fluoroscopic images.

Total knee arthroplasty is now a widely accepted treatment for late-stage arthritis. Wear of the polyethylene layer in prosthetic knees is a known cause of implant failure. Early detection of wear may allow prediction of device failure. In this paper we describe a fully automated image processing algorithm to measure the minimum tibiofemoral joint space width (mJSW) for monitoring prosthesis wear radiographically. The femoral portion and tibial plate were automatically delineated and mJSW was calculated in each compartment. The software also delineated the tip of the prosthesis pin in order to make a magnification correction. The algorithm was tested with a set of triplicate acquisitions of 18 fluoroscopic knee images. The RMS standard deviation (RMSSD) for the triplicate measurements was calculated as a figure of merit. The RMSSD was 0.077 and 0.087 mm for the lateral and medial compartments. The computer successfully found the minimum JSW for both compartments in all 54 images. A single case (2% of total) required user interaction to correct for an obvious failure to delineate the prosthesis pin. We document a robust and precise tool for quantifying mJSW to monitor prosthesis wear.

A radiological technique for the assessment of wear in prosthetic knee replacements.

OBJECTIVE: Wear of the polyethylene insert is a well-recognised cause of implant failure in total knee replacements. The purpose of this study was to evaluate a simple, digital fluoroscopic technique for the assessment of wear in knee prostheses. DESIGN: Fluoroscopic images of knee prostheses were produced both of a phantom and in a patient group. Joint space thickness was measured by reference to a known diameter. Measurements were made to assess repeatability of positioning, inter-and intra-observer variance and the effect of angulation. RESULTS: Standard phantom images showed small variation between measurements, high inter-reader correlation (Pearson's correlation coefficient, r=0.98, P<0.001; coefficient of variation=0.53%) and low intra-reader variation (coefficient of variation=0.57%). Inter- and intra-imager variation were low (coefficient of variation=1.05% and 0.88%, respectively). In the patient group, the range of joint space measurements was 1.9-8.9 mm. The coefficient of variation in insert measurements on repeated images was 2.0%. Repeatability of measurements was 0.2 mm with 99% confidence interval. CONCLUSIONS: This technique allows repeatable, precise measurement of insert thickness. The technique may be adapted to any implant where a reliable calibrating distance is present.

Osteoma of the thyroid cartilage--an unusual cause of difficult intubation.

We describe a case of histologically proven osteoma of the thyroid cartilage that presented because of difficulty in intubation prior to coronary bypass surgery. To our knowledge, this is the first documented case in the English literature.

Giant ancient schwannoma of the pelvis.

A 43-year-old man with a large ancient schwannoma of the pelvis, presenting with varicose veins, is reported. Ancient schwannoma (neurilemmoma) is a benign tumour of nerve sheath origin characterised histologically by features of severe degeneration and which rarely can grow to a large size. Malignant transformation, though reported, is extremely rare.

Extracellular ATP triggers tumor necrosis factor-alpha release from rat microglia.

Brain microglia are a major source of inflammatory cytokines, such as tumor necrosis factor-alpha (TNF-alpha), which have been implicated in the progression of neurodegenerative diseases. Recently, microglia were revealed to be highly responsive to ATP, which is released from nerve terminals, activated immune cells, or damaged cells. It is not clear, however, whether released ATP can regulate TNF-alpha secretion from microglia. Here we demonstrate that ATP potently stimulates TNF-alpha release, resulting from TNF-alpha mRNA expression in rat cultured brain microglia. The TNF-alpha release was maximally elicited by 1 mM ATP and also induced by a P2X(7) receptor-selective agonist, 2'- and 3'-O-(4-benzoylbenzoyl)adenosine 5'-triphosphate, suggesting the involvement of P2X(7) receptor. ATP-induced TNF-alpha release was Ca(2+)-dependent, and a sustained Ca(2+) influx correlated with the TNF-alpha release in ATP-stimulated microglia. ATP-induced TNF-alpha release was inhibited by PD 098059, an inhibitor of extracellular signal-regulated protein kinase (ERK) kinase 1 (MEK1), which activates ERK, and also by SB 203580, an inhibitor of p38 mitogen-activated protein kinase. ATP rapidly activated both ERK and p38 even in the absence of extracellular Ca(2+). These results indicate that extracellular ATP triggers TNF-alpha release in rat microglia via a P2 receptor, likely to be the P2X(7) subtype, by a mechanism that is dependent on both the sustained Ca(2+) influx and ERK/p38 cascade, regulated independently of Ca(2+) influx.

Repeated haloperidol treatment decreases sigma(1) receptor binding but does not affect its mRNA levels in the guinea pig or rat brain.

The effects of chronic treatment with haloperidol on sigma (sigma) receptors were investigated across brain regions and species. The regional distribution of [3H](+)-pentazocine binding to sigma(1) receptor was similar between the guinea pig and rat brains. The highest level of binding was detected in the brain stem and lowest in the striatum and hippocampus. The regional distribution of [3H]1, 3-di (2-tolyl) guanidine ([3H]DTG) binding in the presence of 100 nM (+)-pentazocine to sigma(2) receptor was similar to that of the [3H](+)-pentazocine binding in the guinea pig brain, while in the rat brain high levels of [3H]DTG binding were detected in the cortex, frontal cortex and cerebellum. The intraperitoneal administration of 2 mg/kg of haloperidol to guinea pig and rats once a day for 21 days produced inhibition of [3H](+)-pentazocine binding but did not affect [3H]DTG binding to sigma(2) receptors in any brain region examined. The effects of haloperidol on [3H](+)-penazocine binding in the rat were much weaker than those in the guinea pig. The regional distribution of the level of sigma(1) receptor mRNA determined by the ribonuclease protection assay was similar to that of the [3H](+)-pentazocine binding activity, except in the cortex and cerebellum where the levels of sigma(1) receptor mRNA were low in guinea pig and rat. Treatment with haloperidol did not affect the levels of sigma(1) receptor mRNA in any brain region in either species. These findings suggested that the sigma receptors differentially distributed in brain regions are down regulated by treatment with haloperidol across sigma receptor subtypes and animal species without changing the transcriptional activity of the sigma(1) receptor. The mechanisms by which sigma receptors could be differently regulated in vivo by chronic treatment with haloperidol in different species may contribute to the therapeutic efficacy of haloperidol.

Interleukin-1beta induces substance P release from primary afferent neurons through the cyclooxygenase-2 system.

Substance P (SP) is synthesized in the dorsal root ganglion (DRG) and released from primary afferent neurons to convey information regarding noxious stimuli. The effects of the proinflammatory cytokine interleukin-1 (IL-1) beta on the release of SP were investigated using primary cultured rat DRG cells. Recombinant mouse IL-1beta added to the cells at 0.1 ng/ml increased the SP-like immunoreactivity (SPLI) in the culture medium after incubation for 6 h by approximately 50% as compared with that of nontreated DRG cells. The effect of IL-1beta was Ca(2+)-dependent and significantly inhibited by 100 ng/ml IL-1 receptor-specific antagonist (IL-1r antagonist), cyclooxygenase (COX) inhibitors such as 0.1 mM aspirin, 1 microg/ml indomethacin, and 1 microM NS-398 (specific for COX-2), and 1 microM dexamethasone. Furthermore, a 1-h incubation with IL-1beta markedly increased the inducible COX-2 mRNA level, which was inhibited by an IL-1r antagonist and dexamethasone, whereas IL-1beta showed no effect on the level of constitutive COX-1 mRNA. These observations indicated that IL-1beta induced the release of SP from the DRG cells via specific IL-1 receptors, the mechanism of which might involve prostanoid systems produced by COX-2. This could be responsible for the hyperalgesic action with reference to inflammatory pain in the primary afferent neuron to spinal cord pathway.

Sonographic findings in the anterior interosseous nerve syndrome.

This report details previously undescribed sonographic findings in the anterior interosseous nerve syndrome. Loss of muscle bulk, increased reflectivity, reduced perfusion on Doppler sonography, and lack of active contraction of the affected muscles were observed. These findings can aid in the localization of the pathologic process and in the exclusion of tendon rupture. Dynamic observation of muscle function and Doppler changes after exercise can also help identify the muscles involved. Both sonography and MRI may be useful in the evaluation of patients with the anterior interosseous nerve syndrome and other peripheral neuropathies.