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Matrix-isolation Raman spectroscopy and photochemistry of carbonyl-metal-silyl complexes.

To study the photochemistry of metal-silyl complexes the compounds Cp(CO)(2)Fe-SiH(2)(CH(3)) (Cp = eta(5)-C(5)H(5)) and Cp*(CO)(2)((CH(3))(3)P)Mo-SiH(3) (Cp* = eta(5)-C(5)(CH(3))(5)) have been isolated in krypton or nitrogen matrices and subsequently detected by Raman spectroscopy. Vibrational assignments for the two compounds are given. Furthermore, differences in the Raman spectra induced by UV irradiation are discussed.

Journal Article↗

Subthreshold transpupillary thermotherapy of the retina and experimental choroidal neovascularization in a rat model.

PURPOSE: To study the effect of subthreshold transpupillary thermotherapy (TTT) on the retina and experimental choroidal neovascularization (CNV) in the rat. METHODS: Subthreshold TTT was performed on normal Brown Norway rats or those with krypton laser-induced CNV and appropriate controls with an 810-nm diode laser coupled to a slit lamp. At different intervals after TTT, fundus fluorescence angiography (FFA) and histopathological examinations were performed. Terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL) was used to detect apoptosis. Immunohistochemistry was used to detect heat shock protein 70 (Hsp70) expression. RESULTS: In normal retina, edema and whitening was found on day 1 after TTT. Different degrees of hyperfluorescence could be seen on FFA. Obvious retina damage, especially in the outer layers, was observed by histology in all the lesions that appeared whitened. In the CNV there was congestion and less damage in the overlying retina than in normal retina on day 1 after TTT. Apoptosis was detected in all retinal layers and CNV lesions by TUNEL. In normal eyes, after TTT, Hsp70 expression was increased in the inner layers of the retina and some of the cells in the choroid. Hsp70 was also increased in laser-induced CNV. Two weeks after TTT, the CNV showed a tendency for fibrosis by Masson staining. FFA did not show much change in the CNV lesions 2 weeks after TTT. CONCLUSION: Subthreshold TTT has adverse effects on the overlying retina and thus is likely to cause significant functional and morphological long-term sequelae. Subthreshold TTT can cause apoptosis in laser-induced CNV in rats.

Animals↗

Immunolocalization of transforming growth factor beta during wound repair in rat retina after laser photocoagulation.

BACKGROUND: Scatter photocoagulation induces regression of retinal neovascularization, but the mechanism of its therapeutic effect is incompletely understood. To elucidate the mechanism of therapeutic effect of photocoagulation is the main focus of our research. We have already demonstrated basic fibroblast growth factor (bFGF) immunolocalization during retinal wound repair following laser photocoagulation. Transforming growth factor beta (TGF beta) reportedly inhibits endothelial cell growth and bFGF-induced cell proliferation in vitro. In the present study, we evaluated the immunohistochemical localization of TGF-beta 1 and -beta 2 during wound repair in the rat retina following laser photocoagulation. METHODS: Krypton laser photocoagulation was performed on the eyes of pigmented rats. The eyes were then enucleated on day 1, 3, 7, 14, 28 or 56 following the photocoagulation and enrolled into the analysis of immunohistochemical localization of TGF-beta 1 and -beta 2. RESULTS: Immunoreactivity for TGF-beta 1 and -beta 2 was present in the ganglion cell layer and photoreceptor outer segments of the normal adult rat retina. The cytoplasm of RPE cells at the photocoagulated lesion showed intense TGF-beta 1 and -beta 2 immunoreactivity on day 3 after laser photocoagulation. Macrophages that migrated into the lesion lacked positive staining for TGF-beta 1 and -beta 2. TGF-beta immunoreactivity in RPE cells continued to be upregulated for more than 1 month compared with that in normal RPE cells. Controls did not exhibit any positive staining. CONCLUSION: An elevated expression of TGF-beta immunoreactivity for a longer period of time than bFGF was observed in RPE cells at the photocoagulated lesion in vivo. In the late phase of retinal wound repair, TGF-beta may inhibit cell proliferation induced by mitogens, introduce an end stage of cellular events, and induce extracellular matrix induction.

Animals↗

Fluorescence detection of small gastrointestinal tumours: principles, technique, first clinical experience.

Photodynamic therapy (PDT) is a form of cancer treatment based on the selective accumulation of a photosensitizer in neoplastic tissue. The fluorescent properties of a photosensitizer permit diagnostic localization of primary tumours and/or metastasis. Occult lesions are hard to detect and can easily be missed during routine laparoscopy. Fluorescence observation offers additional optical information and the ability to detect these occult tumours. Clinically, we used 5-aminolevulinic acid for peritoneal staining and tumour demarcation via tumour-specific fluorescence induced by protoporphyrin IX. For laparoscopic observations, a "D-Light" system was used; the conventional white light source was equipped with an optical blocking filter that transmits at the excitation wavelength (380-450 nm) and blocks all other parts of the spectrum. With the aid of a suitable observation filter, the relevant fluorescence was detectable. With the help of this fluorescence we increased the capacity to detect occult tumours, that were missed with white-light observation (9/26). In the gastrointestinal tract, we used a krypton laser at 405 nm for PP IX fluorescence induction. Although there were high sensitivity rates for neoplasms (81% peritoneal carcinomas, 60% gastric cancer), no exact histopathological statement could be achieved at because of false-positive fluorescence, mainly caused by inflammation (6/32). Current clinical goals and the future perspectives of photodynamic diagnostic are discussed.

Aminolevulinic Acid↗

Regional distribution of lung perfusion and ventilation in patients with chronic congestive heart failure and its relationship to cardiopulmonary hemodynamics.

To study the relationship between the distribution of pulmonary blood flow, pulmonary ventilation, and pulmonary capillary wedge pressure, we studied six patients with chronic congestive heart failure (CHF) (New York Heart Association classes II and III) at rest and during exercise. We used krypton 81m (81mKr) and technetium 99m (99mTc) to assess lung ventilation and perfusion at rest and during exercise. Hemodynamic measurements were obtained with a balloon floatation thermodilution catheter. At rest, the upper lung zones of patients with CHF received significantly higher proportions of the blood flow compared with previously published data in normal volunteers. During exercise, however, the fractional perfusion to apices did not change, which suggests that apical flow redistribution is already maximized at rest. Measured pulmonary capillary wedge pressure failed to correlate with the upper zone or the ratio of upper-to-lower zone perfusion counts. Our findings suggest that upper zone flow redistribution may be associated with high, normal, or low capillary wedge pressure and therefore does not correlate with the pulmonary capillary wedge pressure in patients with chronic CHF.

Aged↗

The structure of steroids and their diffusion through blood vessel walls in a counter-current system.

Several substances including prostaglandin F2 alpha, progesterone and 85-krypton have been shown to be transferred from the venous side to the arterial side of the circulation in the ovarian vascular pedicle. Experiments were therefore carried out to study the transfer of three pairs of steroids (progesterone and 20 alpha-dihydroprogesterone, C-21; androstenedione and testosterone, C-19; and estrone and estradiol-17 beta, C-18) in which each member of a pair differed by one hydroxyl group. Each pair of steroids, one labeled with 3H and the other with 14C, were infused in sequence for 30 minutes into a side branch of an ovarian vein near the hilus of the ovary with a rest period of 90 minutes between infusions. An increase in radioactivity in ovarian arterial plasma compared to the radioactivity in an equal volume of aortic plasma sampled simultaneously was used as the index for a direct transfer of steroids from the ovarian vein to the adjacent ovarian artery. All six steroids showed such a transfer which began 3 to 6 minutes after the start of each infusion and decreased rapidly after the infusion was stopped. The results of this study also showed that a larger quantity of the less polar (ketonic) form of each steroid pair examined was transferred than its hydroxyl counterpart.

Androgens↗

The effect of acute hypoxia on right ventricular function in healthy adults.

Right ventricular ejection fraction and right ventricular volumes were derived in 12 healthy male subjects using krypton-81m equilibrium radionuclide ventriculography whilst subjects breathed 30% (high inspired oxygen) and then 8-12% oxygen in nitrogen mixture (hypoxia). 'Physiological' tricuspid valve regurgitation was identified in 7 of the subjects by Doppler echocardiography, and right ventricular peak systolic pressure was estimated during high inspired oxygen and during hypoxia. Mean right ventricular peak systolic pressure was 24.1 +/- 3.3 mmHg during high inspired oxygen and increased to 41.3 +/- 8.4 mmHg during hypoxia (P less than 0.01). Mean right ventricular ejection fraction was 0.612 +/- 0.075 during high inspired oxygen and was unchanged at 0.590 +/- 0.073 during hypoxia. There was no significant change in right ventricular end-diastolic volume or stroke volume in response to hypoxia. The systolic performance of the normal right ventricle is well-maintained during an acute rise in afterload induced by hypoxia.

Acute Disease↗

Separation and detection of acidic and neutral impurities in illicit heroin via capillary electrophoresis.

The separation and detection of acidic and neutral impurities in illicit heroin using capillary electrophoresis (CE) is described. Separations were achieved using charged cyclodextrin modified micellar electrokinetic capillary chromatography. The use of the anionic beta-cyclodextrin sulfobutyl ether 1V in combination with sodium dodecyl sulfate significantly increased resolution. Improved selectivity and/or sensitivity in detection was obtained using photodiode array ultraviolet and laser-induced fluorescence detection. The phenanthrene-like heroin impurities exhibit high native fluorescence under krypton-fluoride laser excitation (248 nm). The limit of detection by laser-induced fluorescence detection for one of these solutes (acetylthebaol) is 1.8 ng/ml, 500 times more sensitive than UV. This methodology is applicable to analysis of both crude and refined heroin.

Drug Contamination↗

Photodynamic therapy with chlorin e6. A morphologic study of tumor damage efficiency in experiment.

Morphological changes in rat sarcoma M-1 after photodynamic treatment with chlorin e6 were studied. The frequency of necrosis appearance and the depth of its spreading in tumor tissue were evaluated after intraperitoneal injection of chlorin e6 in doses of 1-10 mg kg-1 and subsequent irradiation by a krypton laser with light energy density 22.5-135 J cm-2, using the method of vital staining with Evans blue. It was found that the antitumoral effect of photodynamic treatment was strengthened by increasing the dose of the agent and light and reduced by increasing the time interval between chlorin e6 injection and light irradiation. The treatment being given in the parameters mentioned produced a depth of tumor necrosis which varied from 4.0 mm to 16.6 mm. The mechanisms of tumor tissue damage after photodynamic treatment in vivo are discussed.

Animals↗

Recent advances in the synthesis and structure determination of site specifically psoralen-modified DNA oligonucleotides.

We have developed novel methods for the preparation of multimicromole quantities of extremely pure, uniquely photoadducted psoralen-DNA cross-links, furan-side monoadducted DNA and pyrone-side monoadducts. Psoralen cross-linked and furan-side monoadducted DNA were produced by employing high intensity argon ion and krypton ion lasers as light sources. Pyrone-side monoadducts were prepared by base-catalyzed photoreversal of psoralen cross-links. The various psoralen-adducted DNA oligomers were efficiently purified by high performance liquid chromatography. These methods have permitted us to synthesize 4 mumol each of a self-complementary 8-mer d(GCGTACGC) 4'-(hydroxymethyl)-4,5',8-trimethylpsoralen (HMT) furan-side monoadduct and HMT cross-link. Preliminary nuclear magnetic resonance (NMR) data on the HMT cross-linked 8-mer d(GCGTACGC) have been obtained which confirmed the presence of the diadducted psoralen at the unique 5'TpA3' site. NMR data obtained from the 8-mer furan-side monoadduct revealed that the psoralen molecule is intercalated into the DNA double helix. Preliminary crystals of 8-mer cross-linked DNA molecule have been grown. Conditions for the growth of X-ray diffraction-quality crystals and the further analysis of these crystals are now in progress.

Base Sequence↗

Experimental grounds for using chlorin e6 in the photodynamic therapy of malignant tumors.

The toxicity, pharmacokinetics and antitumour effect of chlorin e6 after light irradiation were studied. The LD50 value of chlorin e6 in C3H mice is 189 +/- 9 mg kg-1 and in Wistar white rats is 113 +/- 18 mg kg-1 14 days after intraperitoneal injection. The concentration of chlorin e6 in blood, liver, kidney, spleen and tumors (sarcoma M-1 and sarcoma 45) of the rats was determined by a fluorescence method, 3, 6, 12, 18, 24, 48 and 72 h after administration at a dose of 10 mg kg-1. For this purpose, chlorin e6 was extracted from tissues by the detergent Triton X-100. The depth of necrosis in sarcoma 45, the regression rate of sarcoma M-1 and the animal cure rate were evaluated after chlorin e6 administration at doses of 1-10 mg kg-1 and subsequent irradiation with krypton laser light. Depending on the dose and the time interval between chlorin e6 injection and irradiation, the depth of necrosis in sarcoma 45 varied from 5.0 to 15.0 mm. The cure rate of the animals with sarcoma M-1 varied from 10% to 60%. The antitumor effect was directly proportional to the chlorin e6 dose and light energy exposure and inversely proportional to the time interval between photosensitizer injection and irradiation.

Animals↗

Sensitivity of different rat tumour strains to photodynamic treatment with chlorin e6.

Transplantable rat tumours (sarcoma M-1, sarcoma 45, alveolar liver cancer PC-1 and Pliss' lymphosarcoma) were used to study chlorin e6 accumulation in tumours and its photodynamic effect. Tumours were irradiated by krypton ion laser light (647 and 676 nm; 90 J cm-2) 15 min and 24 h after chlorin e6 injection at doses of 5 and 10 mg kg-1. The relationship between some morphological peculiarities of these tumour strains, photosensitizer accumulation in tumours and their response to the photodynamic treatment is discussed.

Animals↗

Isotopic compositions and accurate masses of single isotopic peaks.

This paper presents and proves an algorithm to calculate the isotopic composition of individual (nominal) isotopic peaks. From this information one can calculate the accurate masses of isotopic peaks. This opens the way to use accurate mass measurements to determine chemical compositions of compounds using non-mono-isotopic peaks. The algorithm is computationally efficient and rigorously correct in the absence of roundoff error. Highly effective error correction strategies are described to detect and correct computational errors arising in practical calculations. Results from theoretical calculations of isotopic masses for a krypton inclusion complex agreed well with experimental measurements.

Algorithms↗

Subthreshold grid laser treatment of macular edema secondary to branch retinal vein occlusion with micropulse infrared (810 nanometer) diode laser.

PURPOSE: To compare the effectiveness of subthreshold grid laser treatment (SGLT) with an infrared micropulse diode laser with that of threshold grid laser treatment (TGLT) for macular edema secondary to branch retinal vein occlusion (BRVO). DESIGN: Randomized clinical trial. PARTICIPANTS: Thirty-six patients (36 eyes) were randomized either to infrared SGLT (17 eyes) or to krypton TGLT (19 eyes). METHODS: Complete ophthalmic examinations, including determination of visual acuity (VA) with Early Treatment Diabetic Retinopathy Study charts, optical coherence tomography (OCT), and fluorescein angiography, were performed at the time of the study entry and at 6-month intervals, with a planned follow-up of 24 months. MAIN OUTCOME MEASURES: Primary: decrease in mean foveal thickness (FT) on OCT. Secondary: changes of the total macular volume (TMV) over the follow-up, proportion of eyes that gained at least 10 letters (approximately > or =2 lines of VA gain) at the 12- and 24-month examinations, and timing of macular edema resolution. RESULTS: Changes in mean FT and TMV from the initial values were statistically significant for TGLT from the 6-month examination (P<0.001) and for SGLT from the 12-month examination (P<0.001). After 1 year, there was no difference in mean FT and TMV between the 2 groups. At the 12-month examination, 10 patients of the SGLT group (59%) and 11 of the TGLT group (58%) gained at least 10 letters (2 lines) in VA. At the 24-month examination, this gain was achieved by 11 patients (65%) of the SGLT group and 11 (58%) of the TGLT group. Moreover, at the 24-month examination 59% and 26% gained 3 lines in the SGLT and TGLT groups, respectively. CONCLUSIONS: Resolution of macular edema and VA improvement are similar to those obtained with conventional TGLT, but SGLT is not associated with biomicroscopic and angiographic signs. A multicenter randomized clinical trial would be needed to ascertain the real efficacy and the most appropriate settings of SGLT for macular edema secondary to BRVO.

Aged↗

Targeted disruption of the FGF2 gene does not prevent choroidal neovascularization in a murine model.

Choroidal neovascularization (CNV) is the major cause of severe visual loss in patients with age-related macular degeneration. Laser treatment is helpful for a minority of patients with CNV, and development of new treatments is hampered by a poor understanding of the molecular signals involved. Several lines of evidence have suggested that basic fibroblast growth factor (FGF2) plays a role in stimulating CNV. In this study, we tested this hypothesis using mice with targeted disruption of the FGF2 gene in a newly developed murine model of laser-induced CNV. One week after krypton laser photocoagulation in C57BL/6J mice, 34 of 60 burns (57%) showed fluorescein leakage and 13 of 16 (81%) showed histopathological evidence of CNV. At 2 weeks, CNV was detected in 9 of 10 burns (90%) in which a bubble had been observed at the time of the laser treatment. Electron microscopy showed fenestrated vessels with large lumens within choroidal neovascular lesions. Two weeks after laser-induced rupture of Bruch's membrane, 27 of 36 burns (75%) contained CNV in FGF2-deficient mice compared with 26 of 30 (87%) in wild-type control mice, a difference that is not statistically significant. This study demonstrates that FGF2 is not required for the development of CNV after laser-induced rupture of Bruch's membrane and provides a new model to investigate molecular mechanisms and anti-angiogenic therapy in CNV.

Animals↗

The thermodynamic response of soft biological tissues to pulsed ultraviolet laser irradiation.

The physical mechanisms that enable short pulses of high-intensity ultraviolet laser radiation to remove tissue, in a process known as laser ablation, remain obscure. The thermodynamic response of biological tissue to pulsed laser irradiation was investigated by measuring and subsequently analyzing the stress transients generated by pulsed argon fluorine (ArF, lambda = 193 nm) and krypton fluorine (KrF, lambda = 248 nm) excimer laser irradiation of porcine dermis using thin-film piezoelectric transducers. For radiant exposures that do not cause material removal, the stress transients are consistent with rapid thermal expansion of the tissue. At the threshold radiant exposure for ablation, the peak stress amplitude generated by 248 nm irradiation is more than an order of magnitude larger than that produced by 193 nm irradiation. For radiant exposures where material removal is achieved, the temporal structure of the stress transient indicates that the onset of material removal occurs during irradiation. In this regime, the variation of the peak compressive stress with radiant exposure is consistent with laser-induced rapid surface vaporization. For 193 nm irradiation, ionization of the ablated material occurs at even greater radiant exposures and is accompanied by a change in the variation of peak stress with radiant exposure consistent with a plasma-mediated ablation process. These results suggest that absorption of ultraviolet laser radiation by the extracellular matrix of tissue leads to decomposition of tissue on the time scale of the laser pulse. The difference in volumetric energy density at ablation threshold between the two wavelengths indicates that the larger stresses generated by 248 nm irradiation may facilitate the onset of material removal. However, once material removal is achieved, the stress measurements demonstrate that energy not directly responsible for target decomposition contributes to increasing the specific energy of the plume (and plasma, when present), which drives the gas dynamic expansion of ablated material. This provides direct evidence that ultraviolet laser ablation of soft biological tissues is a surface-mediated process and not explosive in nature.

Animals↗

Mucosal ablation with photodynamic therapy in the esophagus: optimization of light dosimetry in the sheep model.

BACKGROUND: Photodynamic therapy is an attractive technique for mucosal ablation in patients with superficial squamous cell carcinoma of the esophagus, or high-grade dysplasia or early stage adenocarcinoma arising in Barrett's esophagus. Although illumination with green light is assumed to be safe, choice of the light has been empirical in clinical studies; light dose is often reduced to avoid potential complications. The present study assessed the safety of green and blue lights during photodynamic therapy in the esophagus by progressively administrating increasing doses in an attempt to standardize the dose and determine a safe upper limit. This would considerably simplify photodynamic therapy and improve therapeutic results. METHODS: The sheep model was chosen because of similarities with humans regarding the thickness and histologic structure of the esophagus. Irradiation with a 180 degrees windowed cylindrical light distributor was performed in 19 sheep 4 days after injection of 0.15 mg/kg of tetra(m-hydroxyphenyl) chlorin. Light doses ranged from 10 to 500 J/cm(2) at 514 nm (argon ion laser) and from 5 to 250 J/cm(2) at 413 nm (krypton laser). RESULTS: Follow-up endoscopies revealed a tissue response with a fibrinous area at almost all light doses, whereas application of extremely high light doses tended to induce circumferential necrosis with subsequent stenosis. Three months after irradiation with green light, histologic examination of the resected specimens revealed transmural scarring at doses higher than 100 J/cm(2). After illumination with blue light, partial or more extensive fibrosis of the muscular layer was observed only at light doses of 175 to 250 J/cm(2). CONCLUSIONS: Application of high doses of green light after sensitization with tetra(m-hydroxyphenyl) chlorin led to severe complications in the esophagus of the sheep that are highly likely to occur in humans as well. Blue light causes significantly less damage than green light and may, therefore, be considered as an alternative for photodynamic therapy of early stage superficial esophageal cancer.

Animals↗

A retrospective pilot study of indocyanine green enhanced diode laser photocoagulation for subfoveal choroidal neovascularization associated with age-related macular degeneration.

PURPOSE: The effectiveness and limitations of indocyanine green (ICG) enhanced diode laser photocoagulation in treating subfoveal choroidal neovascularization (CNV) associated with age-related macular degeneration (AMD) were investigated retrospectively. METHODS: Thirty-eight eyes of 37 patients with subfoveal CNV received ICG enhanced diode laser (wavelength, 805 nm) photocoagulation in our preliminary series. Nineteen eyes had classic CNV and the others had occult CNV, which was well-delineated on ICG angiography. The rate of anatomical success and functional outcomes were investigated. Factors prognostic of a final visual acuity of 0.1 or better were analyzed. The follow-up period ranged from 6 to 51 months (mean +/- SD = 26.5 +/- 14.4 months). RESULTS: Occlusion of CNV was achieved in 35 of 38 eyes (92%), and 7 eyes (18%) showed recurrence, which was occluded by retreatment in all but 1 eye. Ten eyes (26.3%) showed improvement of visual acuity; 16 (42.1%) showed no change; and in 12 eyes (31.6%) visual acuity deteriorated. Factors prognostic of a final visual acuity of 0.1 or better were good preoperative visual acuity (Mann-Whitney U-test, P =.0028), and a relatively short distance between the edge of laser burns and the center of the foveal avascular zone (unpaired t-test, P =.0285). CONCLUSION: Indocyanine green enhanced photocoagulation achieved a higher anatomical success rate but functional outcomes equal to those with argon or krypton laser photocoagulation. A controlled prospective study is necessary to prove the efficacy of this treatment.

Aged↗