Characterisation of novel uveal melanoma cell lines under serum-free conditions.

BACKGROUND: The establishment of long-term uveal melanoma (UM) cell lines is difficult. However, studying living cells and their behaviour in the presence of other cells and the extracellular matrix is important in terms of understanding tumour biology and malignant behaviour. We have established three UM cell lines and report a first characterisation of these cell lines. METHODS: Three established UM cell lines (UMT2, UMT26 and UMT33) were analysed according to their morphologic characteristics, melanocytic differentiation, adhesion on different extracellular matrices and proliferative activity. Copy number changes of chromosomes 1, 3, 6 and 8 were studied by multiplex ligation-dependent probe amplification (MLPA). Oncogenic mutations in UM involving exons 4 and 5 of GNAQ and GNA11, respectively, were analysed by sequencing. RESULTS: All cell lines grew in suspension. UMT2 cells were homogeneous, UMT26 and UMT33 cells heterogeneous with regard to cell size and pigmentation. All UM cell lines revealed a melanocytic differentiation. UMT2 and 33 adhered on various extracellular matrices, while UMT26 only adhered to basal membrane extract (BME). This difference corresponded to the different expression of various integrins. Ki67 was expressed by 89% of UMT2 and 95% of UMT33 cells, which thus were in a proliferative stage, while only 2% of UMT26 cells revealed immunostaining for this proliferation marker. The doubling time of UMT2 was 3 days, 12 days for UMT33, and circa 3-4 months for UMT26. MLPA revealed disomy 3 in UMT2 and monosomy 3 in UMT33. The same point mutation was found in UMT2, 26 and 33, in exon 5 of GNA11 at codon 209 (p.Q209L). CONCLUSIONS: The establishment of UM cell lines under serum-free conditions is possible. Characterisation of UMT2, 26, and 33 revealed obvious differences in cytomorphology, melanocytic differentiation, adhesion on extracellular matrices, and proliferative activity. UMT2, 26 and 33 showed the same oncogenic mutation in exon 5 of GNA11.

Dual source multidetector CT-angiography before Transcatheter Aortic Valve Implantation (TAVI) using a high-pitch spiral acquisition mode.

OBJECTIVES: Transcatheter Aortic Valve Implantation (TAVI) is an alternative to surgical valve replacement in high risk patients. Angiography of the aortic root, aorta and iliac arteries is required to select suitable candidates, but contrast agents can be harmful due to impaired renal function. We evaluated ECG-triggered high-pitch spiral dual source Computed Tomography (CT) with minimized volume of contrast agent to assess aortic root anatomy and vascular access. METHODS: 42 patients (82 ± 6 years) scheduled for TAVI underwent dual source (DS) CT angiography (CTA) of the aorta using a prospectively ECG-triggered high-pitch spiral mode (pitch = 3.4) with 40 mL iodinated contrast agent. We analyzed aortic root/iliac dimensions, attenuation, contrast to noise ratio (CNR), image noise and radiation exposure. RESULTS: Aortic root/iliac dimensions and distance of coronary ostia from the annulus could be determined in all cases. Mean aortic and iliac artery attenuation was 320 ± 70 HU and 340 ± 77 HU. Aortic/iliac CNR was 21.7 ± 6.8 HU and 14.5 ± 5.4 HU using 100 kV (18.8 ± 4.1 HU and 8.7 ± 2.6 HU using 120 kV). Mean effective dose was 4.5 ± 1.2 mSv. CONCLUSIONS: High-pitch spiral DSCTA can be used to assess the entire aorta and iliac arteries in TAVI candidates with a low volume of contrast agent while preserving diagnostic image quality.

[Initial experience with prospectively triggered, sequential CT coronary angiography on a 128-slice scanner]. / Erste Erfahrungen mit der sequenziellen, prospektiv getriggerten CT-Koronarangiografie an einem 128-Schicht-Computertomografen.

PURPOSE: Spiral CT angiography (CTA) of the coronaries using low-pitch scanning and ECG-gated image reconstruction is a robust method for detecting or excluding relevant coronary plaque. However, the resulting dose exposure is considerable. The aim of the present study was to evaluate image quality and artifacts as well as to record dose values for sequential coronary CTA using a 128-slice scanner with a temporal resolution of 150 ms. MATERIALS AND METHODS: 20 patients with a regular heart rate and without contraindications for oral/I.V. beta blockers, who were referred for CTA of the coronaries for exclusion or detection of relevant plaques, were examined by sequential CTA with the following parameters: 120 kV, 200 ref mAs, collimation 2 x 64 x 0.6, table feed of 34.5 mm at a detector width of 38.4 mm. A total acquisition time of 380 ms per table position allowed for mild shifting of the reconstruction window within the cardiac cycle of +/- 5 %. 50 ml of contrast agent were injected at 5 ml/s followed by a 50 ml split bolus (20 % contrast). The individual start delay was determined by a test bolus scan (10 ml contrast + 50 ml saline flush at 5 ml/s). The image quality for each segment, coronary artery, and patient was determined on a 4-point scale. Dose values were estimated based on the individual dose length product as provided by the scanner's patient protocol. Artifacts were evaluated to determine the cause (calcium vs. motion). RESULTS: All patients received beta blocker pretreatment. The mean heart rate was 62 +/- 5 beats/min. 5 % (13 / 286) of all segments in 5 / 20 patients were rated as non-diagnostic. The mean dose length product was 213 mGy x cm, and the mean effective dose was 3.6 mSv. Calcifications were the major cause of non-diagnostic images. However breathing or other motion artifacts occurred as well. CONCLUSION: In select patients with effective heart rate control and thorough instruction for breath hold compliance, sequential CTA of the coronaries using a 128-slice scanner with a temporal resolution of 150 ms is technically feasible. The resulting effective dose values are clearly below those of spiral coronary CT scans.