JAK3 deregulation by activating mutations confers invasive growth advantage in extranodal nasal-type natural killer cell lymphoma.

Extranodal, nasal-type natural killer (NK)/T-cell lymphoma (NKCL) is an aggressive malignancy with poor prognosis in which, usually, signal transducer and activator of transcription 3 (STAT3) is constitutively activated and oncogenic. Here, we demonstrate that STAT3 activation mostly results from constitutive Janus kinase (JAK)3 phosphorylation on tyrosine 980, as observed in three of the four tested NKCL cell lines and in 20 of the 23 NKCL tumor samples under study. In one of the cell lines and in 4 of 19 (21%) NKCL primary tumor samples, constitutive JAK3 activation was related to an acquired mutation (A573V or V722I) in the JAK3 pseudokinase domain. We then show that constitutive activation of the JAK3/STAT3 pathway has a major role in NKCL cell growth and survival and in the invasive phenotype. Indeed, NKCL cell growth was slowed down in vitro by targeting JAK3 with chemical inhibitors or small-interfering RNAs. In a human NKCL xenograft mouse model, tumor growth was significantly delayed by the JAK3 inhibitor CP-690550. Altogether, the constitutive activation of JAK3, which can result from JAK3-activating mutations, is a frequent feature of NKCL that deserves to be tested as a therapeutic target.

The Src tyrosine kinase Hck is required for Tel-Abl- but not for Tel-Jak2-induced cell transformation.

Tel-Abl and Tel-Jak2 are fusion proteins associated with human haematologic neoplasms. They possess constitutive tyrosine kinase activity and activate common downstream signalling pathways like Stat-5, PI3-K/Akt, Ras/MapK and NF-kappaB. In this study, we showed the specific requirement of Src family members for the Tel-Abl-mediated cell growth, activation of Stat5, PI3-K/Akt and Ras/MapK while dispensable for Tel-Jak2. Hck was found strongly phosphorylated in Tel-Abl-expressing Ba/F3 cells and sensitive to imatinib mesylate treatment, providing evidence that Hck is a target of Tel-Abl tyrosine kinase activity. Overexpression of a kinase dead form of Hck inhibits the proliferation of Ba/F3 cells expressing Tel-Abl as the phosphorylation of Akt and Erk1/2. These results argue for an important role of Hck in Tel-Abl oncogenic signalling.

[Development and evaluation of a spectroscopy system for classification of laser-induced arterial fluorescence spectra]. / Entwicklung und Erprobung eines Spektroskopiesystems zur Klassifizierung laserinduzierter arterieller Fluoreszenzspektren.

The present study evaluated the potential of fluorescence guidance of laser angioplasty without using a second laser for fluorescence excitation. A prototype spectroscopy system with a grating spectrograph, microchannel plate, CCD array and digital image processor on a personal computer was developed and coupled to a clinical XeCl excimer laser. Using multifibre catheters, specimens of human aorta were ablated in physiological saline and blood. The spectra thus generated were recorded and validated histologically. Five types of spectra could be differentiated. Based on a training set, classification algorithms were developed using multiple linear regression and linear discriminant analysis with intensity ratios as predictor variables. Discriminant analysis yielded prospective classification of the remaining validation spectra with high sensitivity and specificity for each type. These data demonstrate that fluorescence spectroscopy during excimer laser ablation at 308 nm does not require a diagnostic laser. Principal types of atherosclerotic lesions and the media can be differentiated spectroscopically in physiological saline and blood.

Single-laser approach for fluorescence guidance of excimer laser angioplasty at 308 nm: evaluation in vitro and during coronary angioplasty.

BACKGROUND AND OBJECTIVE: Spectroscopic guidance of laser angioplasty has been attempted using a diagnostic He-Cd laser in addition to the therapeutic laser system. This study evaluated a single-laser approach for simultaneous ablation and fluorescence excitation. STUDY DESIGN/MATERIALS AND METHODS: A spectroscopy system was coupled to a clinical XeCl excimer laser. Ablation of 162 human aortic samples in saline and blood with 45 mJ/mm2 per pulse yielded 676 fluorescence spectra validated histologically. The same equipment was used in 16 patients for angioplasty of 18 coronary stenoses applying 500 to 1,725 pulses with 45 to 60 mJ/mm2 under saline flushing. A total of 783 spectra were recorded and validated by intracoronary ultrasound (categories: atheroma, fibrous plaque, calcified lesion). RESULTS: In vitro, 5 types of spectra could be differentiated: (1) atheroma, (2) fibrous plaque, (3) calcified lesion in saline, (4) media, and (5) calcified lesion in blood. Discriminant analysis prospectively classified 576 validation spectra with the following sensitivity and specificity for each type: (1) 83.5 and 97.1%, (2) 85.7 and 96.8% (3) 100 and 98.5%, (4) 98.1 and 99.3%, (5) 98.9 and 100%, respectively. In vivo type 1, 2, 3, and 5 spectra were also observed, but not the media spectrum. The predominant sonographic category also prevailed in spectroscopy. Calcified lesions yielded type 3 and 5 as well as mixed spectra. CONCLUSIONS: Using an excimer laser for angioplasty allows combining ablation and fluorescence excitation without a diagnostic laser. Principal types of atherosclerotic lesions and the media can be differentiated spectroscopically with this approach.

Double-pulse technique for optical emission spectroscopy of ablation plasmas of samples in liquids.

Sharp atomic and ionic line spectra of excimer-laser-induced ablation plasmas of samples in liquids were observed. A new double-pulse technique employing quartz fibers facilitates spectral analysis of samples in liquids in which spectral lines normally are strongly broadened and quenched. The first pulse generates a cavitation bubble on the sample surface, which provides a gaseous environment for optical emission spectroscopy on the ablation plasma induced by a second pulse fired into the bubble.

Ultrafast imaging of tissue ablation by a XeCl excimer laser in saline.

To determine the temporal evolution of laser induced tissue ablation, arterial wall specimens with either hard calcified or fatty plaques and normal tissue were irradiated in a 0.9% saline solution using a XeCl excimer laser (wavelength 308 nm, energy fluence 7 J/cm2, pulse width 30 ns) through a 600 microns fused silica fiber pointing perpendicular either at a 0.5 mm distance or in direct contact to the vascular surface. Radiation of a pulsed dye laser (wavelength 580 nm) was used to illuminate the tissue surface. The ablation process and the arising bubble above the tissue surface were recorded with a CCD camera attached to a computer based image-processing system. Spherical cavitation bubbles and small tissue particles emerging from the irradiated area have been recorded. The volume of this bubble increased faster for calcified plaques than for normal tissue.

In-line holography with a frequency doubled Nd:YAG laser for particle size analysis.

An arrangement for determining the size distribution of small particles (e.g., droplets or bubbles) is presented. It consists of taking high speed holograms with a frequency doubled Nd:YAG laser, reconstructing the real image with an argon-ion laser, and a digital image processing system with a random access image dissector camera and two cascaded computers for filtering, segmentation, and higher recognition tasks. Results are presented for cavitation bubbles as test objects. It is shown that the quality of the holograms is sufficient for detecting bubbles with a diameter of <20 mum by digital image processing with the present configuration.