Ulcerative colitis with overexpression of p53 preceding overt histological abnormalities of the epithelium

Laser-induced fluorescence of pheophorbide a (Ph-a) was used for in vitro photodynamic imaging (PDI) of a rat pancreatic acinar tumor. A 400 nm excitation induced a 470 nm autofluorescence and a 678 nm dye fluorescence in tumors and their surrounding pancreas 24 h after a 9 mg kg-1 body weight Ph-a intravenous administration. With lower intensities in these blood-rich tumors than in pancreas, Ph-a fluorescence signals are unable to provide tumor images. A dimensionless function (the ratio of Ph-a fluorescence by autofluorescence, called Rt for the tumor and Rp for the pancreas) was used for fluorescence contrast calculation (C = Rt/Rp) between six tumors and their paired pancreas. Among five available laser excitation wave-lengths, only the 355 nm excitation gave a distinctive contrast (C = 1.5). The PDI of six intrapancreatic tumors and their intraperitoneal metastasis and of two control normal pancreas was thus performed ex vivo using a 355 nm excitation source delivered by a tripled Nd:YAG laser and a charged-coupled device camera. Fluorescence images were recorded at 680 nm (dye), 640 nm (background) and 470 nm (autofluorescence) through three corresponding 10 nm width bandpass filters. Computed division for each pixel of Ph-a fluorescence values by autofluorescence generated false color image. In this way, contrasted tumor images were obtained. But in five out of six animals false-positive images were present due to an autofluorescence decrease in some normal pancreatic areas. A 470 nm autofluorescence imaging on the same tumors gave in all cases false-positive image and false-negative in half of the cases. These observations suggest that autofluorescence alone is unable to achieve accurate PDI of pancreatic carcinoma and that using Ph-a as a PDI dye needs strong improvements.     

In vivo laser-induced fluorescence imaging of a rat pancreatic cancer with pheophorbide-a

Laser-induced fluorescence (LIF) of pheophorbide-a (Ph-a) was used for imaging of a rat pancreatic tumor. Using a dimensionless function (the ratio of Ph-a fluorescence by bluish autofluorescence), the fluorescence contrasts between excised tumors and their paired pancreas were investigated up to 48 h after a 9 mg kg-1 Ph-a intravenous administration. Among five tested excitation wavelengths, 355 and 610 nm excitations gave the best distinctive contrasts, both 48 h after dye injection. The LIF imaging of six intrapancreatic tumors and six healthy pancreas was carried out in vivo using two laser excitations: 355 nm (Nd:YAG + tripling) for bluish autofluorescence and 610 nm (rhodamine 6G dye) for reddish autofluorescence and dye emission. Images were recorded through bandpass filters at 470 and 640 nm (autofluorescence) and at 680 nm (dye + autofluorescence) with an intensified charged-coupled device camera. Autofluorescence as Ph-a fluorescence images did not allow accurate LIF diagnosis of pancreatic carcinoma. An image processing, including for each pixel a computed division of Ph-a fluorescence (after subtraction of reddish autofluorescence) by bluish autofluorescence intensity generated poorly contrasted tumor images in five of six and false tumor localization in one of three of the tumor-bearing pancreas. A fitting of the digital 640 nm autofluorescence up to the mean 680 nm fluorescence intensity in pancreas prior to subtraction allowed a safe diagnosis to be made with well-contrasted tumor images. To assess automation ability of the processing, a same fitting coefficient (mean of individual values) was applied. In this way, false-negative (one of six) and false-positive (two of six) images were present in tumor-bearing animals as false-positive in one-half of the controls. A successful standardized procedure was then applied with a normalization of 640 and 680 nm pancreas intensities to a same set threshold prior processing. In opposition to thin-layered hollow organs, such as bronchial tube or digestive tract, LIF imaging of carcinoma inserted in a compact organ is exhausting. The use of a dye excitable in the red wavelength range (610 nm for Ph-a) may partly solve this problem, rendering LIF imaging more accurate and potentially automated.     

Psoriatic plaques exhibit red autofluorescence that is due to protoporphyrin IX

In evaluating the autofluorescence properties of normal and diseased skin we discovered that psoriatic plaques can emit a distinct red fluorescence when illuminated with UVA or blue light. Using a macrospectrofluorometer equipped with a 442 nm excitation laser, a sharp in vivo fluorescence emission peak around 635 nm could be demonstrated within the plaques of 34 of 75 (45%) patients with psoriasis. This peak was absent from normal appearing skin of psoriatic patients and also from the skin of 66 patients with other dermatologic diseases. A microspectrofluorometer coupled with the same excitation laser was used to obtain emission spectra of separated epidermal sheets and dermis from plaques demonstrating macroscopic red autofluorescence. An emission peak around 635 nm was observed in all three patients thus studied, but only on spectra obtained from the epidermis. Additional spectra of vertical microscopic sections of intact psoriatic skin from five other patients revealed that the peak originated from the stratum corneum. Emission spectra from other microlocations including the mid-epidermis and dermis of psoriatic and normal skin, as well as the stratum corneum of normal skin, failed to demonstrate a 635 nm peak. The excitation and emission fluorescence spectra of acid extracts of psoriatic scale from five patients were all similar to those of protoporphyrin IX in acid solution. High performance liquid chromatography identified the presence of protoporphyrin IX in the acid extracts from psoriatic scale of the same patients. We conclude that native psoriatic plaques can exhibit red autofluorescence that is due to elevated levels of protoporphyrin IX within scales.     

Studies of the distribution of lipofuscin in the retinal pigment epithelium using high-resolution TV laser scanning ophthalmoscopy

BACKGROUND: The fundus autofluorescence imaging technique has been modified allowing improved image resolution (768 x 572 pixel). We present results of fundus autofluorescence studies using this technique. MATERIALS AND METHODS: Fundus autofluorescence was studied in 286 eyes of 143 patients with retinitis pigmentosa, macular dystrophies and age-related macular degeneration using a confocal laser scanning ophthalmoscope prototype (Zeiss, Oberkochen; excitation wavelength: 488 nm, cut-off filter at 521 nm). RESULTS: The spatial distribution of autofluorescence was different in all diseased eyes investigated compared to the normal pattern of fundus autofluorescence. Each disorder showed a specific fundus autofluorescence appearance. CONCLUSIONS: The advanced technique of imaging fundus autofluorescence allows detailed studies of the lipofuscin distribution. In vivo analysis of the dynamics of accumulation and degradation of lipofuscin in eyes with tapeto-retinal dystrophies and age-related macular disease may contribute to elucidation of the pathogenesis of these disorders.     

Tunable luminescence,energy transfer and excellent thermal stability of SrMg_2(PO_4)_2:Ce~(3+),Tb~(3+) phosphors for LEDs

A series of novel SrMg_2(PO_4)_2:Ce~(3+),Tb~(3+)(SMP:Ce~(3+),Tb~(3+)) phosphors with tunable emission spectra were produced via high temperature solid phase method.XRD,fluorescence spectrum and fluorescence lifetime for SMP:Ce~(3+),Tb~(3+)were studied in detail.Under the excitation at 308 nm,SMP:Ce~(3+),Tb~(3+) samples can emit high efficiency tunable blue-green light by controlling the proportion of dopant concentration.Through the spectral overlap and the regular change of fluorescence lifetime,it is proved that there is a significant energy transfer between Ce~(3+) and Tb~(3+) in SMP matrix and the energy transfer mechanism is determined to be an electric dipole-dipole interaction with energy transmission efficiency of 55%.In additional,Commission International de L'Eclairage(CIE) color coordinates and thermal stability were studied.All above findings suggest that SMP:Ce~(3+),Tb~(3+)can be regarded as the potential bluish green phosphor for LED applications.     

Does heparin coating improve biocompatibility? A study on complement, blood cells and postoperative morbidity during cardiac surgery

BACKGROUND: We investigated fundus autofluorescence in vivo using a novel scanning laser ophthalmoscope. MATERIALS AND METHODS: A total of 550 patients with various retinal diseases were examined and compared with normal eyes. Autofluorescence was detected after excitation with an argon blue laser (488 nm), and emission was recorded with a short wavelength cut off above 500 nm. RESULTS: Reduced autofluorescence was observed in the foveal and parafoveal region due to retinal xanthophyll, along the retinal vessels, at the optic nerve head and in areas with atrophy of the retinal pigment epithelium (RPE). Autofluorescence intensity was increased either focally or diffusely in certain degenerative (AMD) or genetically determined retinal diseases (e.g., Stargardt's disease, Best's disease). CONCLUSIONS: These findings are in accordance with the view that in vivo fundus autofluorescence originates at the level of the RPE and suggest that it is derived from lipofuscin.     

Cytoplasmic viscosity near the cell plasma membrane: measurement by evanescent field frequency-domain microfluorimetry

The purpose of this study was to determine whether the unique physical milieu just beneath the cell plasma membrane influences the rheology of fluid-phase cytoplasm. Cytoplasmic viscosity was evaluated from the picosecond rotation of the small fluorophore 2',7'-bis-(2-carboxyethyl)-5-carboxyfluorescein (BCECF) by parallel-acquisition Fourier transform microfluorimetry (Fushimi and Verkman, 1991). Information about viscosity within < 200 nm of cell plasma membranes was obtained by selective excitation of fluorophores in an evanescent field created by total internal reflection (TIR) of impulse-modulated s-plane-polarized laser illumination (488 nm) at a glass-aqueous interface. Measurements of fluorescence lifetime and time-resolved anisotropy were carried out in solutions containing fluorescein or BCECF at known viscosities, and monolayers of BCECF-labeled Swiss 3T3 fibroblasts and Madin-Darby canine kidney (MDCK) cells. Specific concerns associated with time-resolved fluorescence measurements in the evanescent field were examined theoretically and/or experimentally, including variations in lifetime due to fluorophore proximity to the interface, and the use of the s and p polarized excitation. In fluorescein solutions excited with s-plane polarized light, there was a 5-10% decrease in fluorescein lifetime with TIR compared to trans (subcritical) illumination, but no change in rotational correlation time (approximately 98 ps/cP). Intracellular BCECF had a single lifetime of 3.7 +/- 0.1 ns near the cell plasma membrane. Apparent fluid-phase viscosity near the cell plasma membrane was 1.1 +/- 0.2 cP (fibroblast) and 1.0 +/- 0.2 cP (MDCK), not significantly different from the viscosity measured in bulk cytoplasm far from the plasma membrane. The results establish the methodology for time-resolved microfluorimetric measurement of polarization in the evanescent field and demonstrate that the cell plasma membrane has little effect on the fluid-phase viscosity of adjacent cytoplasm.     

Prediction of suicide intent in aboriginal and non-aboriginal adolescent inpatients: a research note

At 488 nm argon-ion laser excitation human mononuclear cells emit flavoprotein-related autofluorescence signals. Approximately 60% of these are caused by the mitochondrial flavoproteins alpha-lipoamide dehydrogenase and electron transfer flavoprotein, having differences in their fluorescence emission spectra. At the emission wavelength of 530 nm the redox changes of alpha-lipoamide dehydrogenase fluorescence in human mononuclear cells can be monitored by flow cytometry. This allows the estimation of the steady-state reduction level of this flavoprotein being in redox equilibrium with the mitochondrial NAD-system. We applied this method to elucidate the possible impairment of mitochondrial function in subpopulations of mononuclear cells of patients harboring deletions of the mitochondrial DNA in skeletal muscle. In the monocyte fraction of three patients and in the lymphocyte fraction of one patient we observed in the presence of the mitochondrial substrate octanoate elevated steady-state reduction levels of alpha-lipoamide dehydrogenase. This is an indication for the presence of respiratory chain-inhibited mitochondria in mononuclear cell subpopulations of the described patients. These data were confirmed by conventional determinations of maximal oxygen consumption rates of digitonin-permeabilized cells. Therefore, the flow cytometric determination of flavoprotein-caused autofluorescence changes is a useful and sensitive method for the detection of an impairment of mitochondrial respiratory chain in subpopulations of heterogeneous cell suspensions.     

Endogenous skin fluorescence includes bands that may serve as quantitative markers of aging and photoaging

Aging and photoaging cause distinct changes in skin cells and extracellular matrix. Changes in hairless mouse skin as a function of age and chronic UVB exposure were investigated by fluorescence excitation spectroscopy. Fluorescence excitation spectra were measured in vivo, on heat-separated epidermis and dermis, and on extracts of mouse skin to characterize the absorption spectra of the emitting chromophores. Fluorescence excitation spectra obtained in vivo on 6 wk old mouse skin had maxima at 295, 340, and 360 nm; the 295 nm band was the dominant band. Using heat separated tissue, the 295 nm band predominantly originated in the epidermis and the bands at 340 and 360 nm originated in the dermis. The 295 nm band was assigned to tryptophan fluorescence, the 340 nm band to pepsin digestable collagen cross-links fluorescence and the 360 nm band to collagenase digestable collagen cross-links fluorescence. Fluorescence excitation maxima remained unchanged in chronologically aged mice (34-38 wk old), whereas the 295 nm band decreased in intensity with age and the 340 nm band increased in intensity with age. In contrast, fluorescence excitation spectra of chronically UVB exposed mice showed a large increase in the 295 nm band compared with age-matched controls and the bands at 340 and 350 nm were no longer distinct. Two new bands appeared in the chronically exposed mice at 270 nm and at 305 nm. These reproducible changes in skin autofluorescence suggest that aging causes predictable alterations in both epidermal and dermal fluorescence, whereas chronic UV exposure induces the appearance of new fluorphores.     

Upconversion of Er3+ Ions in LiKGdF5∶Er3+, Dy3+ Single Crystal Produced by Infrared and Green Laser

The upconversion fluorescence of Er3 ions in LiKGdF5∶Er3 , Dy3 single crystal was studied under 785, 514.5, and 980 nm laser excitation. With the laser excitation set at 785 nm, strong green (centered at 543 nm) upconversion emissions, as well as weak red (651 nm), violet (406 nm), and blue (470 nm) upconversion emissions were obtained. With 514.5 nm laser excitation, violet (406 nm) and blue (470 nm) upconversion emissions were observed. Under 980 nm laser excitation, strong green (543 nm) and weak red (651) emissions were also obtained. The laser power dependence of the upconverted emissions was investigated to understand the upconversion mechanism. The excited state absorption (ESA) and the energy transfer (ET) processes were discussed as the possible mechanisms for all upconversion emissions.     

Upconversion of Er^3＋ Ions in LiKGdF5 ： Er^3＋ , Dy^3＋ Single Crystal Produced by Infrared and Green Laser

The upconversion fluorescence of Er^3＋ ions in LiKGdF5 ： Er^3＋, Dy^3＋ single crystal was studied under 785, 514.5, and 980 nm laser excitation. With the laser excitation set at 785 nm, strong green （centered at 543 nm） upconversion emissions, as well as weak red （651 nm）, violet （406 nm）, and blue （470 nm） upconversion emissions were obtained. With 514.5 nm laser excitation, violet （406 nm） and blue （470 nm） upconversion emissions were observed. Under 980 nm laser excitation, strong green （543 nm） and weak red （651） emissions were also obtained. The laser power dependence of the upconverted emissions was investigated to understand the upconversion mechanism. The excited state absorption （ESA） and the energy transfer （ET） processes were discussed as the possible mechanisms for all upconversion emissions.     

In situ molecular association of dystrophin with actin revealed by sensitized emission immuno-resonance energy transfer

A novel method was developed to detect molecular associations of dystrophin with actin in cryostat muscle tissue sections by combining resonance energy transfer technology with immunohistochemical techniques. This method takes advantage of the long phosphorescent lifetime of terbium chelates, a property that enables the accurate determination of energy transfer in biological tissues by lifetime measurements of sensitized emission. After a brief excitation pulse, terbium chelates emit for milliseconds after the intrinsically high autofluorescence of biological specimens has decayed to negligible levels. Rat skeletal muscle tissue sections were labeled with both anti-dystrophin monoclonal antibody conjugated to a terbium-based resonance energy transfer donor and anti-actin tetramethylrhodamine phalloidin as an acceptor. Resonance energy transfer between the two probes indicated that the distance separating the probes is within 10 nm (about the size of an IgG2b antibody molecule). The fraction of antibodies that participated in resonance energy transfer was estimated to be 80-90% because of the close agreement between the quenching of donor phosphorescence and the efficiency of resonance energy transfer revealed by lifetime measurements of sensitized emission by tetramethyl-rhodamine phalloidin. Sensitized emission was detectable only when both anti-dystrophin antibody and tetramethyl-rhodamine phalloidin were present. These results indicate that actin and dystrophin are closely associated within the cell. This method is potentially applicable to the investigation of many types of intracellular associations.     

Optical Properties of Dy3+ Doped in Boroaluminasilicate Glass

Dy³⁺ doped boroaluminasilicate glasses were synthesized in air atmosphere with conventional high temperature process. Optical absorption, emission and excitation spectra of the glasses were measured. Effect of concentration quenching on the lifetime and fluorescence yield was discussed. The pilot study on the changes in emission spectra of Dy³⁺/Tb³⁺, Dy³⁺/Pr³⁺ co-doped glasses was carried out. 451 nm was found to be the ideal wavelength among five excitation wavelength in the UV-Vis region from excitation spectra. The fluorescence spectrum of the sample had three major emission bands at 482, 575 and 662 nm, respectively. The strongest emission band peak was at 575 nm. With an increase in Dy³⁺ concentration beyond a particular value (1%), the concentration quenching phenomenon occurred. The lifetime of ⁴F_9/2 level reduced from 0.9 ms to 0.5 ms when Dy³⁺ concentration increased from 0.2% to 4%. The energy transfer from Dy³⁺ to Tb³⁺ and from Pr³⁺ to Dy³⁺ were detected in the co-doped glasses.     

Fluorescence energy-transfer cyanine heterodimers with high affinity for double-stranded DNA. II. Applications to multiplex restriction fragment sizing

Energy-transfer cyanine dyes, a thiazsole orange-thiazole-indolenine (butylTOTIN) and a thiazole orange-thiazole blue heterodimer (pentylTOTAB), form high-affinity complexes with double-stranded (ds)DNA with donor fluorescence (lambdaF(max)527 nm) quenched >90% and with acceptor fluorescence emission above 650 nm (S. C. Benson, Z. Zeng, and A. N. Glaser (1995) Anal. Biochem. 231, 247-255). After separation by agarose gel electrophoresis, bands of precomplexed dsDNA-dye restriction fragments containing 10 pg of dsDNA are readily detected with a laser-excited confocal-fluorescence gel scanner (R. A. Mathies et al. (1994) Rev. Sci. Instrum. 65, 807-812) following donor excitation at 488 nm (argon ion laser) or direct acceptor excitation at 647 nm (krypton ion laser). Accurate two-color multiplex sizing of restriction fragments is obtained with 488-nm excitation with dsDNA fragments precomplexed with thiazole orange dimer (TOTO) as unknowns (detected at 500-565 nm, green channel) and dsDNA fragments stained with the energy-transfer cyanine dyes (detected at 645-750 nm, red channel) as internal standards. There is negligible cross talk of fluorescence between the red and green channels and no significant dye migration in mixtures of prelabeled standard and unknown fragments.     

Functional imaging of mitochondria in saponin-permeabilized mice muscle fibers

Confocal laser-scanning and digital fluorescence imaging microscopy were used to quantify the mitochondrial autofluorescence changes of NAD(P)H and flavoproteins in unfixed saponin-permeabilized myofibers from mice quadriceps muscle tissue. Addition of mitochondrial substrates, ADP, or cyanide led to redox state changes of the mitochondrial NAD system. These changes were detected by ratio imaging of the autofluorescence intensities of fluorescent flavoproteins and NAD(P)H, showing inverse fluorescence behavior. The flavoprotein signal was colocalized with the potentiometric mitochondria-specific dye dimethylaminostyryl pyridyl methyl iodide (DASPMI), or with MitoTrackerTM Green FM, a constitutive marker for mitochondria. Within individual myofibers we detected topological mitochondrial subsets with distinct flavoprotein autofluorescence levels, equally responding to induced rate changes of the oxidative phosphorylation. The flavoprotein autofluorescence levels of these subsets differed by a factor of four. This heterogeneity was substantiated by flow-cytometric analysis of flavoprotein and DASPMI fluorescence changes of individual mitochondria isolated from mice skeletal muscle. Our data provide direct evidence that mitochondria in single myofibers are distinct subsets at the level of an intrinsic fluorescent marker of the mitochondrial NAD-redox system. Under the present experimental conditions these subsets show similar functional responses.     

Optical spectroscopy of nicotinoprotein alcohol dehydrogenase from Amycolatopsis methanolica: a comparison with horse liver alcohol dehydrogenase and UDP-galactose epimerase

The NADH absorbance spectrum of nicotinoprotein (NADH-containing) alcohol dehydrogenase from Amycolatopsis methanolica has a maximum at 326 nm. Reduced enzyme-bound pyridine dinucleotide could be reversibly oxidized by acetaldehyde. The fluorescence excitation spectrum for NADH bound to the enzyme has a maximum at 325 nm. Upon excitation at 290 nm, energy transfer from tryptophan to enzyme-bound NADH was negligible. The fluorescence emission spectrum (excitation at 325 nm) for NADH bound to the enzyme has a maximum at 422 nm. The fluorescence intensity is enhanced by a factor of 3 upon binding of isobutyramide (Kd = 59 microM). Isobutyramide acts as competitive inhibitor (Ki = 46 microM) with respect to the electron acceptor NDMA (N,N-dimethyl-p-nitrosoaniline), which binds to the enzyme containing the reduced cofactor. The nonreactive substrate analogue trifluoroethanol acts as a competitive inhibitor with respect to the substrate ethanol (Ki = 1.6 microM), which binds to the enzyme containing the oxidized cofactor. Far-UV circular dichroism spectra of the enzyme containing NADH and the enzyme containing NAD+ were identical, indicating that no major conformational changes occur upon oxidation or reduction of the cofactor. Near-UV circular dichroism spectra of NADH bound to the enzyme have a minimum at 323 nm (Deltaepsilon = -8.6 M-1 cm-1). The fluorescence anisotropy decay of enzyme-bound NADH showed no rotational freedom of the NADH cofactor. This implies a rigid environment as well as lack of motion of the fluorophore. The average fluorescence lifetime of NADH bound to the enzyme is 0.29 ns at 20 degreesC and could be resolved into at least three components (in the range 0.13-0.96 ns). Upon binding of isobutyramide to the enzyme-containing NADH, the average excited-state lifetime increased to 1.02 ns and could be resolved into two components (0.37 and 1.11 ns). The optical spectra of NADH bound to nicotinoprotein alcohol dehydrogenase have blue-shifted maxima compared to other NADH-dehydrogenase complexes, but comparable to that observed for NADH bound to horse liver alcohol dehydrogenase. The fluorescence lifetime of NADH bound to the nicotinoprotein is very short compared to enzyme-bound NADH complexes, also compared to NADH bound to horse liver alcohol dehydrogenase. The cofactor-protein interaction in the nicotinoprotein alcohol dehydrogenase active site is more rigid and apolar than that in horse liver alcohol dehydrogenase. The optical properties of NADH bound to nicotinoprotein alcohol dehydrogenase differ considerably from NADH (tightly) bound to UDP-galactose epimerase from Escherichia coli. This indicates that although both enzymes have NAD(H) as nonexchangeable cofactor, the NADH binding sites are quite different.     

Judd-Ofelt parameters of the up-conversion phosphors: Er~(3+) doped BaGd_2ZnO_5/PMMA and NaYF_4/PMMA

We proposed a new approach to determination of Judd-Ofelt intensity parameters of Er~(3+) doped phosphors via their absorption spectra. To validate this approach, JO parameters of Er~(3+) doped Ba BaGd_2ZnO_5/PMMA and NaYF_4/PMMA composites were calculated and in a good agreement with the other glass and crystal. The spontaneous radiative transition probability, branching ratio, and radiative lifetime of the optical transitions were calculated by using the Judd-Ofelt theory. Intense near-infrared emission at 1553 nm was observed under 980 nm laser diode excitation at room temperature. The samples possessing high full-width at half-maximum reach 85 nm have potential application in broadband optical amplifier.     

Fabrication and comprehensive structural and spectroscopic properties of Er:Y2O3 transparent ceramics

Transparent Er:Y₂O₃ ceramics with sub-micron grain size (<1 μm) were fabricated by using one-step vacuum sintering followed by hot isostatic pressing (HIPing) technique. The transmission of the undoped Y₂O₃ reaches 83%. The structural characteristics including the phonon energy were investigated through X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR) analysis and scanning electron microscopy (SEM) measurement. The overall spectroscopic properties of transmission, fluorescence emission up to 3000 nm, lifetime, up-conversion luminescence, and refractive index were systematically studied for both 0.25 at% and 7.0 at% Er:Y₂O₃ ceramics with different thicknesses. The comparison of the spectra of the fluorescence emission and up-conversion luminescence under both 976 and 808 nm laser excitation was performed. The multiple high-energy-state transitional processes after the excited state absorption (ESA) processes involved in the up-conversion are discriminated between the multi-phonon non-radiative transitions and the radiative transitions according to the measured maximum phonon vibrational energy. The calculation was performed based on the Judd–Ofelt theory.     

A four-year randomized controlled trial of hormone replacement and bisphosphonate, alone or in combination, in women with postmenopausal osteoporosis

The aim of this study was to investigate the extent of natural intrinsic fluorescence in carious human dentine and any correspondence of such autofluorescence (AF) to the mineral distribution within the lesion. Two investigative techniques were used, both employing the same sample set and fields in diamond-polished block surfaces of polymethylmethacrylate-embedded carious teeth. AF at emission wavelengths >515 nm, excited by 488-nm laser light, was assessed using a confocal laser scanning optical microscope (CLSM), the recordings made under standard operating conditions. The relative mineral content was assessed using digital backscattered scanning electron microscopy (20 kV BSE-SEM). The AF intensity correlated with the level of demineralisation as seen by BSE, but the depth of the lesion as seen by CLSM was significantly greater implying that the AF is not directly related to the mineral component.