Comparison of stone elimination capacity and drilling speed of endoscopic clearance lithotripsy devices.

PURPOSE: To investigate the fragmentation capacity, clearance time, and drilling speed of combined ultrasonic with impact dual-energy and single energy ultrasonic lithotripter devices. METHODS: Stone fragmentation and clearance tests were performed under direct view in an underwater layered hemisphere by four different operators using artificial stones (n = 10/operator). Time for complete clearance was measured. Drilling tests were performed using an underwater setup, consisting of a mounting rack for fixing the lithotripter handpiece with the probe in vertical position and in contact with the stone phantom placed on one side of a balance for defined and constant contact application pressure equivalent to 450 g load. Time until complete perforation or in case of no perforation, the penetration depth after 60 s into the stone sample was recorded. Four devices, one single energy device (SED), one dual-energy dual probe (DEDP), two dual-energy single probe (DESP-1, DESP-2), with different parameters were tested. RESULTS: Stone fragmentation and clearance speed were significantly faster for dual-energy device DESP-1 compared to all other devices (p < 0.001). Using DESP-1, the clearance time needed was 26.0 ± 5.0 s followed by DESP-2, SED and DEDP requiring 38.4 ± 5.8 s, 40.1 ± 6.3 s and 46.3 ± 11.6 s, respectively. Regarding the drilling speed, DESP-1 was faster compared to all other devices used (p < 0.05). While the drilling speed of DESP-1 was 0.69 ± 0.19 mm/s, compared to 0.49 ± 0.18 mm/s of DESP-2, 0.47 ± 0.09 mm/s of DEDP, and 0.19 ± 0.03 mm/s of SED. CONCLUSIONS: The dual-energy/single-probe device combining ultrasonic vibrations with electromechanical impact was significantly faster in fragmentation and clearing stone phantoms as well as in drilling speed compared to all other devices.

In vitro investigations of propulsion during laser lithotripsy using video tracking.

OBJECTIVES: Ureteroscopic laser lithotripsy is an important and widely used method for destroying ureter stones. It represents an alternative to ultrasonic and pneumatic lithotripsy techniques. Although these techniques have been thoroughly investigated, the influence of some physical parameters that may be relevant to further improve the treatment results is not fully understood. One crucial topic is the propulsive stone movement induced by the applied laser pulses. To simplify and speed up the optimization of laser parameters in this regard, a video tracking method was developed in connection with a vertical column setup that allows recording and subsequently analyzing the propulsive stone movement in dependence of different laser parameters in a particularly convenient and fast manner. MATERIALS AND METHODS: Pulsed laser light was applied from below to a cubic BegoStone phantom loosely guided within a vertical column setup. The video tracking method uses an algorithm to determine the vertical stone position in each frame of the recorded scene. The time-dependence of the vertical stone position is characterized by an irregular series of peaks. By analyzing the slopes of the peaks in this signal it was possible to determine the mean upward stone velocity for a whole pulse train and to compare it for different laser settings. For a proof of principle of the video tracking method, a specific pulse energy setting (1 J/pulse) was used in combination with three different pulse durations: short pulse (0.3 ms), medium pulse (0.6 ms), and long pulse (1.0 ms). The three pulse durations were compared in terms of their influence on the propulsive stone movement in terms of upward velocity. Furthermore, the propulsions induced by two different pulse energy settings (0.8 J/pulse and 1.2 J/pulse) for a fixed pulse duration (0.3 ms) were compared. A pulse repetition rate of 10 Hz was chosen for all experiments, and for each laser setting, the experiment was repeated on 15 different freshly prepared stones. The latter set of experiments was compared with the results of previous propulsion measurements performed with a pendulum setup. RESULTS: For a fixed pulse energy (1 J/pulse), the mean upward propulsion velocity increased (from 120.0 to 154.9 mm · s-1 ) with decreasing pulse duration. For fixed pulse duration (0.3 ms), the mean upward propulsion velocity increased (from 91.9 to 123.3 mm · s-1 ) with increasing pulse energy (0.8 J/pulse and 1.2 J/pulse). The latter result corresponds roughly to the one obtained with the pendulum setup (increase from 61 to 105 mm · s-1 ). While the mean propulsion velocities for the two different pulse energies were found to differ significantly (P < 0.001) for the two experimental and analysis methods, the standard deviations of the measured mean propulsion velocities were considerably smaller in case of the vertical column method with video tracking (12% and 15% for n = 15 freshly prepared stones) than in case of the pendulum method (26% and 41% for n = 50 freshly prepared stones), in spite of the considerably smaller number of experiment repetitions ("sample size") in the first case. CONCLUSION: The proposed vertical column method with video tracking appears advantageous compared to the pendulum method in terms of the statistical significance of the obtained results. This may partly be understood by the fact that the entire motion of the stones contributes to the data analysis, rather than just their maximum distance from the initial position. The key difference is, however, that the pendulum method involves only one single laser pulse in each experiment run, which renders this method rather tedious to perform. Furthermore, the video tracking method appears much better suited to model a clinical lithotripsy intervention that utilizes longer series of laser pulses at higher repetition rates. The proposed video tracking method can conveniently and quickly deliver results for a large number of laser pulses that can easily be averaged. An optimization of laser settings to achieve minimal propulsive stone movement should thus be more easily feasible with the video tracking method in connection with the vertical column setup. Lasers Surg. Med. 50:333-339, 2018. © 2017 Wiley Periodicals, Inc.

Laser-assisted fixation of a nitinol stapes prosthesis.

BACKGROUND AND OBJECTIVE: Otosclerosis is an inner ear bone disease characterized by fixation of the stapes and consequently progressive hearing loss. One treatment option is the surgical replacement of the stapes by a prosthesis. When so called "smart materials" like nitinol are used, prosthesis fixation can be performed using a laser without manual crimping on the incus. However, specific laser-prosthesis interactions have not been described yet. The aim of the present study was to elucidate the thermo-mechanical properties of the NiTiBOND® prosthesis as a basis for handling instructions for laser-assisted prosthesis fixation. STUDY DESIGN AND MATERIALS AND METHODS: Closure of the NiTiBOND® prosthesis was induced ex vivo by either a diode laser emitting at λ = 940 nm or a CO2 laser (λ = 10,600 nm). Total energy for closure was determined. Suitable laser parameters (pulse duration, power per pulse, distance between tip of the laser fiber and prosthesis) were assessed. Specific laser-prosthesis interactions were recorded. RESULTS: Especially the diode laser was found to be an appropriate energy source. A total energy deposit of 60 mJ by pulses in near contact application was found to be sufficient for prosthesis closure ex vivo. Energy should be transmitted through a laser fiber equipollent to the prosthesis band diameter. Specific deformation characteristics due to the zonal prosthesis composition have to be taken into account. CONCLUSION: NiTiBOND® stapes prosthesis can be closed by very little energy when appropriate energy sources like diode lasers are used, suggesting a relatively safe application in vivo. Lasers Surg. Med. 50:153-157, 2018. © 2017 Wiley Periodicals, Inc.

Impact of pulse duration on Ho:YAG laser lithotripsy: fragmentation and dusting performance.

OBJECTIVES: In vitro investigations of Ho:YAG laser-induced stone fragmentation were performed to identify potential impacts of different pulse durations on stone fragmentation characteristics. MATERIALS AND METHODS: A Ho:YAG laser system (Swiss LaserClast, EMS S.A., Nyon, Switzerland) with selectable long or short pulse mode was tested with regard to its fragmentation and laser hardware compatibility properties. The pulse duration is depending on the specific laser parameters. Fragmentation tests (hand-held, hands-free, single-pulse-induced crater) on artificial BEGO stones were performed under reproducible experimental conditions (fibre sizes: 365 and 200 µm; laser settings: 10 W through combinations of 0.5, 1, 2 J/pulse and 20, 10, 5 Hz, respectively). RESULTS: Differences in fragmentation rates between the two pulse duration regimes were detected with statistical significance for defined settings. Hand-held and motivated Ho:YAG laser-assisted fragmentation of BEGO stones showed no significant difference between short pulse mode and long pulse mode, neither in fragmentation rates nor in number of fragments and fragment sizes. Similarly, the results of the hands-free fragmentation tests (with and without anti-repulsion device) showed no statistical differences between long pulse and short pulse modes. CONCLUSION: The study showed that fragmentation rates for long and short pulse durations at identical power settings remain at a comparable level. Longer holmium laser pulse duration reduces stone pushback. Therefore, longer laser pulses may result in better clinical outcome of laser lithotripsy and more convenient handling during clinical use without compromising fragmentation effectiveness.

Impact of pulse duration on Ho:YAG laser lithotripsy: treatment aspects on the single-pulse level.

OBJECTIVES: Holmium-YAG (Ho:YAG) laser lithotripsy is a multi-pulse treatment modality with stochastic effects on the fragmentation. In vitro investigation on the single-pulse-induced effects on fiber, repulsion as well as fragmentation was performed to identify potential impacts of different Ho:YAG laser pulse durations. MATERIALS AND METHODS: A Ho:YAG laser system (Swiss LaserClast, EMS S.A., Nyon, Switzerland) with selectable long- or short-pulse mode was tested with regard to fiber burn back, the repulsion capacity using an underwater pendulum setup and single-pulse-induced fragmentation capacity using artificial (BEGO) stones. The laser parameters were chosen in accordance with clinical application modes (laser fiber: 365 and 200 µm; output power: 4, 6 and 10 W in different combinations of energy per pulse and repetition rate). Evaluation parameters were reduction in fiber length, pendulum deviation and topology of the crater. RESULTS: Using the long-pulse mode, the fiber burn back was nearly negligible, while in short-pulse mode, an increased burn back could be observed. The results of the pendulum test showed that the deviation induced by the momentum of short pulses was by factor 1.5-2 higher compared to longer pulses at identical energy per pulse settings. The ablation volumes induced by single pulses either in short-pulse or long-pulse mode did not differ significantly although different crater shapes appeared. CONCLUSION: Reduced stone repulsion and reduced laser fiber burn back with longer laser pulses may result in a more convenient handling during clinical application and thus in an improved clinical outcome of laser lithotripsy.

Laser lithotripsy of salivary stones: Correlation with physical and radiological parameters.

BACKGROUND AND OBJECTIVES: Sialolithiasis is a common disease of the major salivary glands. Owing to the variety of conservative and minimally invasive techniques, it is now possible to treat most cases of sialolithiasis without removal of the affected salivary gland. One treatment option is the endoscopic removal of the calculi. In cases of larger concretions, intraductal disintegration using laser-induced shock waves can be appropriate to allow endoscopic removal. In the present study, we investigated whether physical and radiological parameters of salivary stones can effectively predict the applicability of laser lithotripsy. Furthermore, we determined to what extent the applied laser energy resulted in tissue damage. STUDY DESIGN/MATERIALS AND METHODS: In addition to basic parameters like size and density, we analysed 47 salivary stones using fluorescence spectroscopy, infrared spectroscopy, Raman spectroscopy, and dual-energy computed tomography. Subsequent fragmentation of all stones was performed with a Ho:YAG laser in a near-contact manner. Fragmentation rates were calculated and correlated with the previously measured physical and radiological parameters. Finally, to test for tissue damage, we performed HE-histology of salivary duct mucosa treated with the same laser energy used for stone fragmentation. RESULTS: Blue light excitation induced either green or red fluorescence emission. Dual-energy CT resulted in evidence of calcium-containing material. Infrared spectroscopy and Raman spectroscopy, both identified carbonate apatite as the main component of salivary stones. Disintegration into pieces smaller than 2 mm was possible in all cases. Fragmentation rates depended on the energy per pulse applied but not on any of the analysed physical and radiological parameters. In contrast to lithotripsy with 500 mJ per pulse, which was associated with no tissue damage, lithotripsy with 1,000 mJ per pulse resulted in damage of salivary duct mucosa. This suggests that the optimal laser energy for stone fragmentation is between 500 and 1,000 mJ per pulse. CONCLUSION: Laser lithotripsy using Ho:YAG laser is a highly efficient treatment, at least in vitro. All salivary stones could be disintegrated irrespective of their physical and radiological composition.

In vitro investigations of repulsion during laser lithotripsy using a pendulum set-up.

Ureteroscopic laser lithotripsy is a commonly used technique to treat ureteral calculi.The type of energy source used is one of the main influences of retrograd calculi propulsion. Using a momentum pendulum under-water set-up the induced momentum and the initial velocity were investigated. Pulsed laser light from three different clinically available laser systems, including a Ho:YAG laser, a frequency-doubled double-pulse (second harmonic generation, SHG) Nd:YAG laser and a flash-lamp pumped dye (FLPD) laser, were transmitted via flexible fibres of different core diameter to the front of the pendulum sinker. Single pulses at variable pulse energy, according to the clinical laser parameter settings, were applied to the target sinker, thus causing a repulsion-induced deflection which was documented by video recording. The maximum deflection was determined. Solving the differential equation of a pendulum gives the initial velocity, the laser-induced momentum and the efficiency of momentum transfer. The induced deflection as well as the starting velocity of the two short-duration pulsed laser systems (SHG Nd:YAG, FLPD) were similar (s (max) = 2-3.6 cm and v (0) = 150-200 mm/s, respectively), whereas both values were lower using the Ho:YAG laser with a long pulse duration (s (max) = 0.9--1.6 cm and v (0) = 60-105 mm/s, respectively). The momentum I induced by the Ho:YAG laser was only 50% and its transfer efficacy η (Repuls) was reduced to less than 5% of the values of the two short-pulsed laser systems. This investigation clearly showed the variable parts and amounts of repulsion using different pulsed lasers in an objective and reproducible manner. The momentum transfer efficiency could be determined without any physical friction problems. Further investigations are needed to compare stone fragmentation techniques with respect to laser repulsion and its clinical impact.

Comparative investigation of reusable and single-use flexible endoscopes for urological interventions.

In order to evaluate the technical adaptability of a type of disposable endoscope compared to reusable flexible endoscopes, in vitro and in vivo studies were conducted. A disposable digital ureteroscope ("chip on tip") and two reusable endoscopes were investigated with respect to spatial resolution, geometric distortion in air and water the maximum. Additionally, the clinical performance of the disposable device was tested during clinical procedures (n = 20). The disposable endoscope showed an optical resolution of 6.72 lines/mm at 10 mm distance, similar to the other devices. In comparison, the disposable endoscope showed a barrel-shaped image distortion in air of -24.2%, which is in the middle range, but was best under water (-8.6%). The bendability of 297° (275 µm fiber) and 316° (empty channel, 1.5 F basket) and the maximum irrigation (1 m: 58.1 ml/min, 2 m: 91.9 ml/min) were convincing. Clinically the maneuverability was very good in (13/20), good or satisfactory in (7/20). Visibility was evaluated as very good in (11/20), just in (1/20) either satisfactory or sufficient. The consistency of visibility was not affected in (19/20). In all cases there were no adverse events. The technical examination and clinical application of the disposable endoscope are of equal quality compared to reusable devices. Disposable endoscopes can be an alternative to reusable devices, but economic aspects such as reduction of repair costs, sterilization effort and additional waste must be taken into account.

MOP-dependent enhancement of methadone on the effectiveness of ALA-PDT for A172 cells by upregulating phosphorylated JNK and BCL2.

BACKGROUND: Methadone, as a long-acting opioid analgesic, shows an ability to sensitize the treatment of ALA-PDT for glioblastoma cells (A172) in vitro by promoting apoptosis. However, the mechanisms how methadone enhances the effectiveness of ALA-PDT for tumor cells remains to be clarified. METHODS: The expression of mu opioid receptor (MOP), apoptosis, phosphorylated c-Jun N-terminal kinase (JNK) and phosphorylated apoptosis regulator B cell lymphoma 2 (BCL2) were measured by flow cytometry. Cytotoxicity was determined using Cell Counting Kit-8 (CCK-8). A MOP antagonist, naloxone, was used to evaluate the role of MOP in the above process. RESULTS: It was found that A172 cells show the expression of MOP and that naloxone inhibits the enhancement of the methadone effect on apoptosis following ALA-PDT (p < 0.05). Phosphorylated JNK and BCL2 induced by ALA-PDT were promoted in the presence of methadone (p < 0.05). These methadone effects were also inhibited by naloxone (p < 0.05). CONCLUSIONS: The results suggest that apoptosis induced by ALA-PDT is enhanced by methadone, mostly MOP-mediated, through the upregulation of accumulation of phosphorylated JNK and BCL2, leading to a promotion of cytotoxicity of ALA-PDT for A172 cells.

Methadone enhances the effectiveness of 5-aminolevulinic acid-based photodynamic therapy for squamous cell carcinoma and glioblastoma in vitro.

Although having shown promising clinical outcomes, the effectiveness of 5-aminolevulinic acid-based photodynamic therapy (ALA-PDT) for squamous cell carcinoma (SCC) and glioblastoma remains to be improved. The analgesic drug methadone is able to sensitize various tumors to chemotherapy. In this in vitro study, the influence of methadone to the effectiveness of ALA-PDT for SCC (FADU) and glioblastoma (A172) was investigated on the protoporphyrin IX (PpIX) fluorescence, survival rates, apoptosis, and cell cycle phase, each with or without the presence of methadone. The production of PpIX was increased by methadone in FADU cells while it was decreased in A172 cells. The survival rates of both cell lines treated by ALA-PDT were significantly reduced by the combination with methadone (P < .05). Methadone also significantly increased the percentage of apoptotic cells and improved the effect of ALA-PDT on the cell cycle phase arrest in the G0/G1 phase (P < .05). This study demonstrates the potential of methadone to influence the cytotoxic effect of ALA-PDT for both SCC and glioblastoma cell lines.

Holmium:yttrium-aluminum-garnet laser induced lithotripsy: in-vitro investigations on fragmentation, dusting, propulsion and fluorescence.

The fragmentation efficiency on Bego artificial stones during lithotripsy and the propulsive effect (via video tracking) was investigated for a variety of laser settings. A variation of the laser settings (pulse energy, pulse duration, repetition rate) altered the total application time required for stone fragmentation, the stone break up time, and the propulsion. The obtained results can be used to develop lithotripsy devices providing an optimal combination of low stone propulsion and high fragmentation efficacy, which can then be evaluated in a clinical setting. Additionally, the fluorescence of human kidney stones was inspected endoscopically in vivo. Fluorescence light can be used to detect stone-free areas or to clearly distinguish calculi from surrounding tissue or operation tools.

Investigation of optical properties of dissected and homogenized biological tissue.

Knowledge of tissue optical properties, in particular the absorption µa and the reduced scattering coefficient µs', is required for diagnostic and therapeutic applications in which the light distribution during treatment has to be known. As it is generally very difficult to obtain this information with sufficient accuracy in vivo, optical properties are often approximately determined on ex vivo tissue samples. In this case, the obtained optical properties may strongly depend on the sample preparation. The extent of the expectable preparation-dependent differences was systematically investigated in comparative measurements on dissected and homogenized porcine tissue samples (liver, lung, brain, and muscle). These measurements were performed at wavelengths 520, 635, 660, and 785 nm, using a dual-step reflectance device and at a spectral range of 515 to 800 nm with an integrating sphere setup. In a third experiment, the density of tissue samples (dissected and homogenized) was investigated, as the characteristic of the packaging of internal tissue structures strongly influences the absorption and scattering. The standard errors of the obtained absorption and reduced scattering coefficients were found to be reduced in case of homogenized tissue. Homogenizing the tissues also allows a much easier and faster sample preparation, as macroscopic internal tissue structures are destroyed in the homogenized tissue so that a planar tissue sample with well-defined thickness can easily and accurately be prepared by filling the tissue paste into a cuvette. Consequently, a better reproducibility result was obtained when using homogenized samples. According to the density measurements accomplished for dissected and homogenized tissue samples, all types of tissues, except lung, showed a decrease in the density due to the homogenization process. The presented results are in good agreement for µs' regardless of the preparation procedure, whereas µa differs, probably influenced by blood content and dehydration. Because of faster and easier preparation and easier sample positioning, homogenization prior to measurement seems to be suitable for investigating the optical properties ex vivo. Additionally, by means of using the homogenization process, the sample size and thickness do not need to be particularly large, as is the case for most biopsies from the OR.

ALA and malignant glioma: fluorescence-guided resection and photodynamic treatment.

BACKGROUND: Oral application of 20 mg/kg body weight of 5-aminolevulinic acid (ALA) leads to a highly specific accumulation of fluorescent Protoporphyrin IX (PPIX) in malignant glioma tissue. In the past few years, we have participated in several clinical studies designed to investigate fluorescence guided resection (FGR) and photodynamic therapy (PDT). METHODS: PPIX selectivity and PPIX bleaching during PDT were assessed with spectroscopic measurements. FGR was performed in 18 clinics in Germany (ALA-Glioma Study Group, participants see end of paper) in a phase III trial comprising an ALA group and a white-light group. PDT was performed with microlens fibers or cylindrical diffusers postsurgically to the resection bed. Additionally, a protocol for the interstitial stereotactic placement of cylindrical diffusers was established and applied on patients with recurrent, inoperable glioblastoma. RESULTS: Compared to normal cortex, mean PPIX fluorescence in vital tumor was found more than 100-fold increased. During PDT, the PPIX fluorescence bleached to 8%, 16%, and 1% of the initial intensity for the 100, 150, and 200 J/cm2 groups (median values). FGR: Contrast-enhancing tumor was completely resected in 65% of patients in the ALA group compared to 36% in the white-light group (p < 0.0001). Progression-free survival was superior in the ALA group compared to white-light patients with cumulative 6 months progression-free survival rates of 41% and 21% (p = 0.0003), respectively. Interstitial PDT can be performed with multiple radial diffusers approximately 10 mm apart, 200 mW/cm, and an irradiation time of one hour.

In-vitro investigations on laser-induced smoke generation mimicking the laparoscopic laser surgery purposes.

Intraoperative smoke-generation limits the quality of vision during laparoscopic/endoscopic laser-assisted surgeries. The current study aimed at the evaluation of factors affecting this phenomenon. As a first step, a suitable experimental setup and a test tissue model were established for this investigation. The experimental setup is composed of a specific sample container, a laser therapy component suitable for the ablation of model tissue at different treatment wavelengths (λ = 980 nm, 1350 nm, 1470 nm), a suction unit providing continuous smoke extraction, and a detection unit for smoke quantification via detection of light (λ = 633 nm) scattered from smoke particles. The ablation rate (AR) was calculated by dividing the ablated volume by the ablation time (60 sec). The laser-induced scattering signal intensity of the smoke (SI) was determined from time-charts of the signal intensity as a measure for vision, in addition a delay-time tdelay could be derived defining the onset of SI after the laser was switched on. The ratio SI/AR is used as a measure for smoke generation in relation to the ablation rate. Additionally the light transmission of the tissue samples was used to estimate their optical properties. In this set-up, smoke generation using λ = 980 nm as ablation laser wavelength was detected after a delay-time tdelay = (121.6 ± 24.8) sec which is significantly longer compared to the wavelengths λ = 1350 nm with tdelay = (89.8 ± 19.3) sec and λ = 1470 nm with tdelay = (24.7 ± 5.4) sec. Thus, the delay Experimental set-up consisting of sample container, laser therapy component, suction unit and scattered-light detection compartment. time is wavelength-dependent. The SI/AR ratio was significantly different (p < 0.001) for 1470 nm irradiation compared to 980 nm irradiation [SI/AR(1470) = (11.8 ± 2.6) · 10(3) vs. SI/AR(980) = (8.6 ± 2.0) · 10(3) ]. The ablation crater for 980 nm irradiation was comparable with 1470 nm irradiation, but the coagulation rim was thicker in the 980 nm case. In conclusion, it could be shown experimentally that smoke-generation depends on the wavelength used for laser ablation.

Association of small dense LDL serum levels and circulating monocyte subsets in stable coronary artery disease.

OBJECTIVE: Atherosclerosis is considered to be an inflammatory disease in which monocytes and monocyte-derived macrophages play a key role. Circulating monocytes can be divided into three distinct subtypes, namely in classical monocytes (CM; CD14++CD16-), intermediate monocytes (IM; CD14++CD16+) and non-classical monocytes (NCM; CD14+CD16++). Low density lipoprotein particles are heterogeneous in size and density, with small, dense LDL (sdLDL) crucially implicated in atherogenesis. The aim of this study was to examine whether monocyte subsets are associated with sdLDL serum levels. METHODS: We included 90 patients with angiographically documented stable coronary artery disease and determined monocyte subtypes by flow cytometry. sdLDL was measured by an electrophoresis method on polyacrylamide gel. RESULTS: Patients with sdLDL levels in the highest tertile (sdLDL≥4mg/dL;T3) showed the highest levels of pro-inflammatory NCM (15.2±7% vs. 11.4±6% and 10.9±4%, respectively; p<0.01) when compared with patients in the middle (sdLDL=2-3mg/dL;T2) and lowest tertile (sdLDL=0-1mg/dL;T1). Furthermore, patients in the highest sdLDL tertile showed lower CM levels than patients in the middle and lowest tertile (79.2±8% vs. 83.9±7% and 82.7±5%; p<0.01 for T3 vs. T2+T1). Levels of IM were not related to sdLDL levels (5.6±4% vs. 4.6±3% vs. 6.4±3% for T3, T2 and T1, respectively). In contrast to monocyte subset distribution, levels of circulating pro- and anti-inflammatory markers were not associated with sdLDL levels. CONCLUSION: The atherogenic lipoprotein fraction sdLDL is associated with an increase of NCM and a decrease of CM. This could be a new link between lipid metabolism dysregulation, innate immunity and atherosclerosis.

In vitro comparisons of retropulsion and fragmentation efficacy of 2 cordless, handheld pneumatic and electromechanical lithotripsy devices.

OBJECTIVE: To compare, in vitro, probe velocity/displacement, retropulsion, and fragmentation capacity of the cordless electromechanical (LithoBreaker) (hard vs soft probe guide) and pneumatic (StoneBreaker). MATERIALS AND METHODS: Probe velocities/displacements were measured using high-speed resolution camera (100.000 frames/s). The lithotripsy probes were projected through a 7.5F ureteroscope against a nonfrangible led ball placed in a 15F horizontally mounted silicone tube immersed in water bath as an in vitro ureter model. Retropulsion is considered as displacement distance of led ball. Fragmentation efficiency was quantified as number of shots required to break Bego-stone phantoms (hard [15:3] and soft [15:6], average size 7.5 × 5.5 mm) placed on metal mesh into <3-mm fragments. Mean and standard deviation of repetitive measurements were statistically analyzed. RESULTS: StoneBreaker yielded higher probe velocity (22 ± 1.9 m/s) compared with LithoBreaker with hard (14.2 ± 0.5 m/s) and soft (11.5 ± 0.5 m/s) probe. Maximum probe displacement for StoneBreaker was 1.04 mm vs 0.9 mm and 1.1 mm (hard vs soft LithoBreaker-probe, respectively). Retropulsion using 1-mm probes showed no statistical differences. Using harder 2-mm probe decreased Lithobreaker retropulsion significantly compared with Stonebreaker. The amount of shots (1-mm probe) to fragment soft Bego stones was significantly higher for LithoBreaker with soft (mean 31.5 ± 11.31) and hard (mean 21.5 ± 5.29) probe guide vs StoneBreaker (mean 11.2 ± 2.65). Fragmentation efficiency for hard Bego stones showed similar statistically significant outcome. Comparison of the 2 probe guides showed higher velocity linked to harder-probe that improved LithoBreaker fragmentation performance and reduce propulsion. CONCLUSION: Both examined lithotripters are effective in cracking stone phantoms with relatively low pulse number. They produce comparable retropulsions. Fragmentation improved substantially using LithoBreaker with hard probe guide. More tests are required to assess differences in stone clearance time.

Small high-density lipoprotein is associated with monocyte subsets in stable coronary artery disease.

OBJECTIVE: High-density lipoprotein (HDL) particles are heterogeneous in structure and function and the role of HDL subfractions in atherogenesis is not well understood. It has been suggested that small HDL may be dysfunctional in patients with coronary artery disease (CAD). Monocytes are considered to play a key role in atherosclerotic diseases. Circulating monocytes can be divided into three subtypes according to their surface expression of CD14 and CD16. Our aim was to examine whether monocyte subsets are associated with HDL subfractions in patients with atherosclerosis. METHODS: We included 90 patients with angiographically stable CAD. Monocyte subsets were defined as classical monocytes (CD14++CD16-; CM), intermediate monocytes (CD14++CD16+; IM) and non-classical monocytes (CD14+CD16++; NCM). HDL subfractions were measured by electrophoresis on polyacrylamide gel. RESULTS: Serum levels of small HDL correlated with circulating pro-inflammatory NCM and showed an inverse relationship to circulating CM independently from other lipid parameters, risk factors, inflammatory parameters or statin treatment regime, respectively. IM were not associated with small HDL. In particular, patients with small HDL levels in the highest tertile showed dramatically increased levels of NCM (14.7 ± 7% vs. 10.7 ± 5% and 10.8 ± 5%; p = 0.006) and a decreased proportion of CM (79.3 ± 7% vs. 83.7 ± 6% and 83.9 ± 6%; p = 0.004) compared to patients in the two lower tertiles. In contrast, intermediate HDL, large HDL and total HDL were not associated with monocyte subset distribution. CONCLUSION: Small HDL levels are associated with pro-inflammatory NCM and inversely correlated with CM. This may suggest that small HDL could have dysfunctional anti-inflammatory properties in patients with established CAD.

Photodynamic therapy for cholangiocarcinoma using low dose mTHPC (Foscan(®)).

BACKGROUND: Photodynamic therapy (PDT) combined with stenting is an effective treatment modality for palliation of nonresectable cholangiocarcinoma (CC). A drawback of standard PDT using Photofrin(®) as photosensitizer is the long lasting skin photosensitivity of up to 3 months. The aim of this study was to show the outcome of PDT of CC, potential side effects and to determine the best drug light interval (DLI) using mTHPC (Foscan(®)) at a low dose. METHODS: 13 patients with nonresectable CC were treated with stenting and PDT (3mg Foscan(®) per treatment, 0.032-0.063 mg/kg body weight, 652 nm, 50 J/cm). Fluorescence measurements were performed with a single bare fiber for 5/13 patients prior to PDT at the tumor site to determine the fluorescence contrast. For another 7/13 patients, long-term fluorescence-kinetics were measured on the oral mucosa to determine the time of maximal relative fluorescence intensity. RESULTS: The results so far indicate a median survival time of 13 months. Side effects such as perforations or skin phototoxicity could not be observed. Foscan(®) fluorescence within the tumor site was clearly detectable but a significant fluorescence contrast of tumor to adjacent healthy tissue could not be found. The fluorescence kinetics measured in the oral mucosa showed a maximum at 3.85 days (median) after drug administration. CONCLUSION: Combined stenting and PDT performed with a low Foscan(®) dose results in equal and potentially longer survival times compared to standard Photofrin(®) PDT, while lowering the risk of side effects strongly. Thus it may improve the quality of life.

Photodynamic therapy of bladder cancer - a phase I study using hexaminolevulinate (HAL).

OBJECTIVES: To assess the safety and feasibility of hexaminolevulinate (HAL) based photodynamic therapy (PDT) as adjuvant treatment after transurethral resection of the bladder (TURB) in patients with intermediate or high-risk urothelial cell carcinoma (UCC) of the bladder. MATERIALS AND METHODS: Seventeen patients received 50 ml of either a 16 mM (4 patients) or 8 mM HAL (13 patients) solution instilled intravesically. Bladder wall irradiation was performed using an incoherent white light source coupled via a quartz fiber assembled into a flexible transurethral irrigation catheter. Each patient received 3 treatments with HAL-PDT 6 weeks apart. After PDT, patients were followed by regular cystoscopy for up to 21 months to assess time to recurrence. Reported adverse events (AEs) were coded according the World Health Organization Adverse Reaction Terminology (WHO-ART). Efficacy was assessed by cystoscopy, cytology, and histology, and was defined as the number of patients who were tumor-free at 6 or 21 months after initial PDT treatment. Transient bladder irritability was reported by 15 of the 17 patients and resolved completely in all patients. No evidence of a cumulative effect of treatment on the incidence of AEs could be detected. PDT treatment was performed without any technical complications. Furthermore preliminary assessment of efficacy showed that of the 17 patients included, 9 (52.9%; 95% CI: 27.8-77.0) were tumor-free at 6 months, 4 (23.5%; 95% CI: 6.8-49.9) were tumor-free at 9 months, and 2 (11.8%, 95% CI: 1.5-36.4) were tumor-free after 21 months. CONCLUSIONS: PDT using hexaminolevulinate and an incoherent white light system with the special flexible irradiation catheter system is technically feasible and safe and may offer an alternative in the treatment of non-muscle-invasive intermediate and high-risk bladder cancer.

Induction of immune mediators in glioma and prostate cancer cells by non-lethal photodynamic therapy.

BACKGROUND: Photodynamic therapy (PDT) uses the combination of photosensitizing drugs and harmless light to cause selective damage to tumor cells. PDT is therefore an option for focal therapy of localized disease or for otherwise unresectable tumors. In addition, there is increasing evidence that PDT can induce systemic anti-tumor immunity, supporting control of tumor cells, which were not eliminated by the primary treatment. However, the effect of non-lethal PDT on the behavior and malignant potential of tumor cells surviving PDT is molecularly not well defined. METHODOLOGY/PRINCIPAL FINDINGS: Here we have evaluated changes in the transcriptome of human glioblastoma (U87, U373) and human (PC-3, DU145) and murine prostate cancer cells (TRAMP-C1, TRAMP-C2) after non-lethal PDT in vitro and in vivo using oligonucleotide microarray analyses. We found that the overall response was similar between the different cell lines and photosensitizers both in vitro and in vivo. The most prominently upregulated genes encoded proteins that belong to pathways activated by cellular stress or are involved in cell cycle arrest. This response was similar to the rescue response of tumor cells following high-dose PDT. In contrast, tumor cells dealing with non-lethal PDT were found to significantly upregulate a number of immune genes, which included the chemokine genes CXCL2, CXCL3 and IL8/CXCL8 as well as the genes for IL6 and its receptor IL6R, which can stimulate proinflammatory reactions, while IL6 and IL6R can also enhance tumor growth. CONCLUSIONS: Our results indicate that PDT can support anti-tumor immune responses and is, therefore, a rational therapy even if tumor cells cannot be completely eliminated by primary phototoxic mechanisms alone. However, non-lethal PDT can also stimulate tumor growth-promoting autocrine loops, as seen by the upregulation of IL6 and its receptor. Thus the efficacy of PDT to treat tumors may be improved by controlling unwanted and potentially deleterious growth-stimulatory pathways.