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Nm Light (nm + light)
Selected AbstractsAn in vivo comparison of photoactivatable fluorescent proteins in an avian embryo modelDEVELOPMENTAL DYNAMICS, Issue 6 2007Danny A. Stark Abstract Tracing the lineage or neighbor relationships of cells in a migratory population or deep within an embryo is difficult with current methods. The recent explosion of photoactivatable fluorescent proteins (PAFPs) offers a unique cell labeling tool kit, yet their in vivo performance in intact embryos and applicability have not been thoroughly explored. We report a comparison study of PAGFP, PSCFP2, KikGR, and Kaede analyzed in the avian embryo using confocal and 2-photon microscopy. PAFPs were introduced into the chick neural tube by electroporation and each photoconverted in the neural crest or cells in the neural tube with exposure to 405 nm light, but showed dramatic differences in photoefficiency and photostability when compared at the same 2% laser power. KikGR and Kaede photoconverted with ratios only slightly lower than in vitro results, but cells rapidly photobleached after reaching maximal photoefficiency. PSCFP2 had the lowest photoefficiency and photoconverted nearly 70 times slower than the other dual-color PAFPs tested, but was effective at single-cell marking, especially with 2-photon excitation at 760 nm. The dual-color PAFPs were more effective to monitor cell migratory behaviors, since non-photoconverted neighboring cells were fluorescently marked with a separate color. However, photoconverted cells were limited in all cases to be visually distinguishable for long periods, with PSCFP2 visible from background the longest (48 hr). Thus, photoactivation in embryos has the potential to selectively mark less accessible cells with laser accuracy and may provide an effective means to study cell,cell interactions and short-term cell lineage in developmental and stem cell biology. Developmental Dynamics 236:1583,1594, 2007. © 2007 Wiley-Liss, Inc. [source] Fluorescence Spectroscopy of Color Centers Generated in Phosphate Glasses after Exposure to Femtosecond Laser PulsesJOURNAL OF THE AMERICAN CERAMIC SOCIETY, Issue 5 2002James W. Chan A confocal fluorescence microscopy setup was used to observe, in situ, spectral changes in phosphate glasses which were modified using 0.3 ,J of tightly focused 800 nm, 130 fs laser pulses. On 488 nm excitation, the modified glass shows a broad fluorescence centered at roughly 600 nm, which decays with prolonged exposure to the 488 nm light. The decay behavior is dependent on the 488 nm power, with a faster decay rate for higher powers. A mechanism whereby color centers, formed by the femtosecond pulses, fluoresce when excited by the 488 nm light and are simultaneously photobleached is proposed to explain the observed behavior. [source] Visible light-induced killing of bacteria as a function of wavelength: Implication for wound healingLASERS IN SURGERY AND MEDICINE, Issue 6 2010Anat Lipovsky MSc Abstract Background and objective Visible light (400,800,nm) at high intensity was previously found to kill bacteria that are frequently found in infected wounds, while low-power white light enhances bacterial proliferation. The phototoxic effect was found to involve induction of reactive oxygen species (ROS) production by the bacteria. The aim of the present study was to identify the most effective wavelengths in the visible range for inducing a bactericidal effect. Experimental ROS production in Staphylococcus aureus and Escherichia coli as a function of wavelengths in the visible range (400,500, 500,800, 415, and 455,nm) was studied using the electron paramagnetic resonance (EPR) spin trapping technique. The phototoxicity of 415 and 455,nm light at different fluencies on the survival of S. aureus and E. coli was assessed by colony count of the bacteria following irradiation. Results ROS production following blue (400,500,nm) light illumination was found to be higher than that of red (500,800,nm). Within the blue range, light of 415,nm induced more ROS than 455,nm, which correlated with results obtained for the reduction in colony count of S. aureus and E. coli following illumination using equal intensities of these two wavelengths. At low fluencies, both 415 and 455,nm enhanced proliferation of S. aureus but reduced viability of E. coli. Conclusion Intense blue light, preferably at 415,nm, could be used for bacterial eradication. However, it should be noted that low intensity of visible light can be dangerous since it may promote proliferation of the microorganisms. Lasers Surg. Med. 42:467,472, 2010. © 2010 Wiley,Liss, Inc. [source] Amelioration of Airway Stenosis in Rabbit Models by Photodynamic Therapy with Talaporfin Sodium (NPe6)PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 3 2009Yoshinori Nakagishi It is difficult to treat patients with acquired airway stenosis, and the quality of life of such patients is therefore lowered. We have suggested the application of photodynamic therapy (PDT) as a new treatment for airway stenosis and have determined the efficacy of PDT in animal disease models using a second-generation photosensitizer with reduced photosensitivity. An airway stenosis rabbit model induced by scraping of the tracheal mucosa was administered NPe6 (5 mg kg,1), and the stenotic lesion was irradiated with 670 nm light emitted from a cylindrical diffuser tip at 60 J cm,2 under bronchoscopic monitoring. PDT using NPe6 improved airway stenosis (P = 0.043) and respiratory stridor. A significant prolongation of survival time was seen in the PDT-treated animals compared to that in the untreated animals (P = 0.025) and 44% of the treated animals achieved long-term survival (>60 days). In conclusion, PDT using NPe6 is effective for improvement in airway stenosis. [source] Fluorescence Resonance Energy Transfer Between Polyphenolic Compounds and Riboflavin Indicates a Possible Accessory Photoreceptor Function for Some Polyphenolic CompoundsPHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 5 2006Kumar Chandrakuntal ABSTRACT The photoreceptive extreme tip of the wheat coleoptile exhibits intense green-yellow fluorescence under UV light, suggesting the presence of UV-absorbing materials. Fluorescence spectra of the intact coleoptile tip and tip homogenate showed the presence of the known photoreceptor pigments flavin and carotene, and a preponderance of phenolic compounds. Absorption spectra and fluorescence spectra of various phenolic compounds showed close overlap with the absorption and fluorescence spectra of the wheat coleoptile tip homogenate. Fluorescence spectra of several phenolic compounds showed close overlap with the absorption bands of flavin, carotene and pterine, suggesting possible energy transduction from phenols to these photoreceptors. Excitation of gentisic acid and ferulic acid with 340 nm light in the presence of flavin showed enhancement of flavin fluorescence in a concentration- and viscosity-dependent fashion, indicating fluorescence resonance energy transfer between them and riboflavin. Furthermore, several phenolic compounds tested generated superoxide anion on excitation at 340 nm, suggesting that superoxide-dependent signal cascades could operate in a polyphenol-mediated pathway. Phenolic compounds thus may act as accessory photoreceptors bringing about excitation energy transfer to the reactive photoreceptor molecules, or they may take over the function of the normal photoreceptor in genetic mutations lacking the system, or both processes may occur. The responses of plants to UV-B and UV-A light in mutants may be explained in terms of various phenolics acting as energy transducers in photoreceptor functioning. [source] The Laser Two-photon Photolysis of Liquid Carbon Tetrachloride,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 1 2006G. Zhang ABSTRACT The two-photon photolysis of liquid CCl4 with 25 ps pulses of 266 nm light has been studied and compared with similar studies with high energy radiation. Both neutral and ionic species are produced from excited states and ionization. The emphasis of the study is on the ionic processes, while some data related to excited states and free radicals are presented. In both radiolysis and photolysis, a solvent separated charged pair, CCl3+, Cl - , exhibiting a ,max at 475 nm, is observed that exhibits a total growth over 38 to 100 ps. Solutes with ionization potentials less than that of CCl4 (11.47 eV) reduce the yield of the 475 nm species producing radical cations of the solute. The efficiency of this process is about 10-fold larger in radiolysis compared with photolysis. Analysis of the data suggest that the lower energy of two-photon photolysis produces a charge pair CCl4+, CCl4 - , which decays in about 3 ps to CCl4+, Cl - . This species then decays to CCl3+, Cl - . The lifetime of the growth of the 475 nm is measured as 46 ps. These studies clearly show areas where radiolysis and photolysis can be quite similar and also areas where the vast difference in excitation energy introduces stark differences in the observed radiation and photoinduced chemistry. [source] Calculation of Singlet Oxygen Dose from Photosensitizer Fluorescence and Photobleaching During mTHPC Photodynamic Therapy of MLL Cells,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 1 2005Jonathan S. Dysart ABSTRACT Predicting the therapeutic outcome of photodynamic therapy (PDT) requires knowledge of the amount of cytoxic species generated. An implicit approach to assessing PDT efficacy has been proposed where changes in photosensitizer (PS) fluorescence during treatment are used to predict treatment outcome. To investigate this, in vitro experiments were performed in which Mat-LyLu cells were incubated in meta -tetra(hydroxyphenyl)chlorin (mTHPC) and then irradiated with 652 nm light. PS concentration, fluence rate and oxygenation were independently controlled and monitored during the treatment. Fluorescence of mTHPC was monitored during treatment and, at selected fluence levels, cell viability was determined using a colony-formation assay. Singlet oxygen dose was calculated using four different models and was compared with cell survival. For the dose metric based on singlet oxygen,mediated PS photobleaching, a universal relationship between cell survival and singlet oxygen dose was found for all treatment parameters. Analysis of the concentration dependence of bleaching suggests that the lifetime of singlet oxygen within the cell is 0.05,0.25 ,s. Generation of about 9 × 108 molecules of singlet oxygen per cell reduces the surviving fraction by 1/e. [source] Cationic Dye,sensitized Degradation of Sodium Hyaluronate Through Photoinduced Electron Transfer in the Upper Excited State,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 3 2001Masanobu Kojima ABSTRACT The formation of ground-state complexes of methylene blue (MB) and thionine (TN) with sodium hyaluronate (NaHA) was clearly observed by means of absorption spectra in aqueous solution. Irradiation of the complexes using 313 nm light caused significant degradation of NaHA under oxygen and argon. However, the use of visible light over 400 nm, which gives the lowest excited singlet state of the cationic dyes, caused no degradation. MB and TN were more efficient sensitizers for the degradation of NaHA than rose bengal (RB), although RB is a more efficient singlet oxygen (1O2) sensitizer than the cationic dyes. Under similar conditions the polysaccharides with carboxyl groups, such as alginic acid and polygalacturonic acid, also photodecomposed. However, the polysaccharides without carboxyl groups, such as pullulan and methyl cellulose, did not. The irradiation of the polysaccharides in the presence of powdered titanium dioxide as a photocatalyst to generate the hydroxyl radical (·OH) in aerated aqueous solution caused the fragmentation of all the polymers. It was confirmed that methyl viologen, an electron-accepting sensitizer, formed a charge-transfer complex with NaHA, the irradiation of which caused the efficient degradation of NaHA. In the presence of ,- and ,-cyclodextrins the MB- and TN-sensitized photodegradation of NaHA was markedly suppressed. This was probably due to the formation of the inclusion complexes comprising the cationic dyes and the cyclodextrins. On the basis of the results obtained we propose that the cationic dye,sensitized degradation of NaHA involves a photoinduced electron-transfer process between the upper excited dyes and the ground-state NaHA and that ·OH and 1O2 do not participate in the degradation. [source] NMDA potentiation by visible light in the presence of a fluorescent neurosteroid analogueTHE JOURNAL OF PHYSIOLOGY, Issue 12 2009Lawrence N. Eisenman N -Methyl- d -aspartate (NMDA) receptors are widely studied because of their importance in synaptic plasticity and excitotoxic cell death. Here we report a novel method of potentiating NMDA receptors with fluorescence excited by blue (480 nm) light. In the presence of 300 nm of a (7-nitro-2,1,3-benzoxadiazol-4-yl) amino (NBD)-tagged neuroactive steroid carrier C2-NBD-(3,,5,)-3-hydroxypregnan-20-one (C2-NBD 3,5,P), responses of cultured hippocampal neurons to 10 ,m NMDA were potentiated to 219.2 ± 9.2% of the baseline response (100%) by a 30 s exposure to 480 nm light. The potentiation decayed back to baseline with a time constant of 80.6 s. Responses to 1 ,m and 100 ,m NMDA were potentiated to 147.9 ± 9.6% and 174.1 ± 15.6% of baseline, respectively, suggesting that visible-light potentiation is relatively insensitive to NMDA concentration. Peak autaptic NMDA responses were potentiated to 178.9 ± 22.4% of baseline. Similar potentiation was seen with 10 ,m NBD-lysine, suggesting that visible-light potentiation is not a steroid effect. Potentiation was also seen with a steroid analogue in which the NBD was replaced with fluorescein, suggesting that NBD is not the only fluorophore capable of supporting visible-light potentiation. UV light and redox potentiation of NMDA receptors largely occluded subsequent blue light potentiation (127.7 ± 7.4% and 120.2 ± 6.2% of baseline, respectively). The NR1a(C744A,C798A) mutant that is insensitive to redox and UV potentiation was also largely unaffected by visible-light potentiation (135.0 ± 10.0% of baseline). Finally, we found that the singlet oxygen scavenger furfuryl alcohol decreased visible-light potentiation. Collectively, these data suggest that visible-light potentiation of NMDA receptors by fluorescence excitation shares mechanisms with UV and redox potentiation and may involve singlet oxygen production. [source] Synovial ablation in a rabbit rheumatoid arthritis model using photodynamic therapyANZ JOURNAL OF SURGERY, Issue 7 2002Andrew D. Beischer Background: At present there is no ideal minimally invasive method for ablating inflamed synovium in joints that has been unresponsive to optimal medical management in patients with rheumatoid arthritis. The aim of this study was to determine whether photodynamic therapy could be used for this purpose. Methods: In a rabbit knee model of rheumatoid arthritis the pharmacokinetics of the photosensitizer Haematoporphyrin Derivative (HpD) into periarticular tissues and blood was measured following intravenous injection of HpD. The second phase of the study was to determine the histological effect of HpD activation by 63 nm light delivered via an intra-articular optic fibre using a dye pumped KTP-YAG laser. The light dose was varied from 0,200 joule/cm2. Results: Pharmacokinetic studies determined that inflamed synovium rapidly accumulated HpD, with peak levels being reached 12 h following intravenous injection. The ratio of HpD uptake into inflamed synovium versus peri-articular quadriceps muscle was found to be 22.8. Histological examination of the treated knees indicated that selective destruction of inflamed synovium was achieved at light doses 100 joules/cm2 and above. No significant effect was observed on normal intra-articular tissues. Conclusion: We have demonstrated that the first generation photosensitizer HpD selectively accumulates within inflamed synovium. Activation of HpD by intra-articular light administration resulted in selective ablation of the inflamed synovium. These findings indicate that PDT offers potential as a new selective, minimally invasive synovectomy technique. [source] Caged Protein Prenyltransferase Substrates: Tools for Understanding Protein PrenylationCHEMICAL BIOLOGY & DRUG DESIGN, Issue 3 2008Amanda J. DeGraw Originally designed to block the prenylation of oncogenic Ras, inhibitors of protein farnesyltransferase currently in preclinical and clinical trials are showing efficacy in cancers with normal Ras. Blocking protein prenylation has also shown promise in the treatment of malaria, Chagas disease and progeria syndrome. A better understanding of the mechanism, targets and in vivo consequences of protein prenylation are needed to elucidate the mode of action of current PFTase (Protein Farnesyltransferase) inhibitors and to create more potent and selective compounds. Caged enzyme substrates are useful tools for understanding enzyme mechanism and biological function. Reported here is the synthesis and characterization of caged substrates of PFTase. The caged isoprenoid diphosphates are poor substrates prior to photolysis. The caged CAAX peptide is a true catalytically caged substrate of PFTase in that it is to not a substrate, yet is able to bind to the enzyme as established by inhibition studies and X-ray crystallography. Irradiation of the caged molecules with 350 nm light readily releases their cognate substrate and their photolysis products are benign. These properties highlight the utility of those analogs towards a variety of in vitro and in vivo applications. [source] | |||||||||||||