The localization of catecholamine fluorescence to dog hypothalamic neuromelanin-bearing neurons.
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Fiber-optic probes are widely used in optical spectroscopy of biological tissues and other turbid media. Only limited information exists, however, on the ways in which the illumination-collection geometry and the overall probe design influence the interrogation of media. We have investigated both experimentally and computationally the effect of probe-to-target distance (PTD) on the diffuse reflectance collected from an isotropically (Lambertian) scattering target and an agar-based tissue phantom. Studies were conducted with three probes characterized by either common (single-fiber) or separate (two bifurcated multifiber probes) illumination and collection channels. This study demonstrates that PTD, probe design, and tissue scattering anisotropy influence the extent of the transport of light into the medium, the light-collection efficiency, and the sampling volume of collected light. The findings can be applied toward optimization of fiber-optic probe designs for quantitative optical spectroscopy of turbid media including biological tissues.
We have developed a tapping-mode scanning near-field optical Raman microscope (SNORM) with a caved and pyramidical probe, using resonant Raman scattering, and have measured the stress distribution of Si. The peak frequency shifts to a lower frequency by 0-0.5 cm(-1) in the area covered by silicon dioxide, whereas it shifts to a higher frequency by 0-0.3 cm(-1) in the area uncovered by silicon dioxide, showing that the areas covered and uncovered by silicon dioxide are under tensile and compressive stresses, respectively. It has been found that compressive stresses of about 0.69 GPa/cm2 are concentrated on the corner of the area uncovered by silicon dioxide. The comparison of stress distributions measured with and without the cantilever shows that the SNORM we developed has a spatial resolution of at least less than 250 nm.
Ex vivo lung-stage larvae of Schistosoma mansoni and S. haematobium do not bind specific antibodies in the indirect membrane immunofluorescence test (IF), probably as a result of confinement of the surface membrane antigens in immobile, lipid-rich sites. Treatment with the membrane-impermeable, cholesterol-extracting drug methyl-beta-cyclodextrin (MBCD) and staining with filipin III (filipin), a fluorescent polyene antibiotic widely used for the detection and quantitation of cholesterol in biomembranes, allowed us to examine the role of cholesterol in surface membrane antigen sequestration of S. mansoni and S. haematobium ex vivo lung-stage larvae. Treatment of S. mansoni larvae with MBCD elicited appreciable cholesterol depletion as judged by filipin-cholesterol fluorescence diminution, which was accompanied by a considerable increase in specific antibody binding in IF, thus suggesting that cholesterol plays a predominant role in sequestration of the surface membrane antigens of S. mansoni lung-stage schistosomula. Despite that, MBCD induced an almost complete depletion of cholesterol from the outer membrane of S. haematobium larvae; no increase in specific antibody binding in IF was evident, implying that cholesterol is not responsible for masking surface membrane antigens of S. haematobium lung-stage larvae.
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BACKGROUND: The question of whether the repair of an alveolar bony defect can be enhanced by orthodontic tooth movement was addressed. METHODS: Alveolar bone defects were created in 52 Wistar male rats anterior to both maxillary first molars. After 1 week of healing, orthodontic protraction was applied for 2 weeks on the right side, resulting in mesial tipping and displacement movement. Subsequently, a retention appliance was inserted for 1 week. The left side served as the untreated (control) group. Vital bone staining (procion brilliant red H-8) was administered before and after orthodontic traction. Histomorphometric analysis was performed on 62 hemimaxillae using UV confocal microscopy and an imaging program. The total area of the bony defect was divided into 4 equal quadrants, and the area of bony apposition in each quadrant was measured. RESULTS: The total area of bony apposition was 6.5-fold larger in the treated (26.41 x 10(4) +/- 28.92 x 10(4) microm2) than in the control group (4.07 x 10(4) +/- 2.82 x 10(4) microm2), approaching statistical significance (P = 0.065). The treated occlusal quadrants demonstrated highly significant (P= 0.010), greater bone apposition compared to the control group (13.8-fold) and to the treated apical quadrants (P= 0.04, 5-fold). CONCLUSIONS: This study confirms that orthodontic tooth movement is a stimulating factor of bone apposition. A conversion in the repair pattern of the bony defect from apicoocclusal in the control group (no tooth movement) to occlusoapical in the treated group (with tooth movement) further supports the linkage between tooth movement and enhanced bone deposition. Clinical implication suggests incorporation of orthodontic tooth movement in regenerative therapy.
Didemnum molle is a colonial ascidian that harbors the prokaryotic photosymbiont Prochloron in its cloacal cavity. Colonies occur over a relatively wide bathymetric range (approximately 0-30 m), and colony color is widely variable, partly depending on depth. Colonies in shallow sites are bright white, with densely distributed spicules, and often with brown or dark gray pigmentation, while colonies in deeper sites are less pigmented, with sparsely distributed spicules. Didemnum molle colonies contain mycosporine-like amino acids (MAAs) as UV-absorbing substances. These include mycosporine-glycine, shinorine, and porphyra-334. Among colonies from 5-, 10-, 15-, and 20-m depths, the concentration of total MAAs was significantly high at 10 m and low at 20 m. Colonies at 10 m need to maintain low spicule densities to have enough photosynthetically active radiation (PAR) to maintain the photosymbionts, and they probably concentrate MAAs to block UV radiation without attenuating PAR. Because high levels of PAR cause photoinhibition of photosynthesis, spicules and pigment cells would be more effective for photoprotection in shallow water. Colonies of D. molle may adjust the light conditions for photosymbionts by combining MAAs, spicules, and pigment cells in varying amounts.
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Identification of the strains controlling bone remodeling is important for determining ways to prevent bone loss due to load deprivation, or implant placement. Long-term monitoring of strains can potentially provide the best information. Glues are resorbed within 2-3 weeks. Two formulations of microcrystalline hydroxyapatite (HA) were used to attach strain gages to rat femora to assess their long-term in vivo strain measurement capability. Seven male rats received HA-coated gages, and 2 animals underwent a sham procedure. The gages were prepared using a published technique and placed on the antero-lateral aspect of the left femora. After 6-7 weeks, the animals were euthanized and both femora explanted. Gages were attached to the right femora with cyanoacrylate. All femora were tested in cantilever bending, then embedded, sectioned, and stained with mineralized bone stain. The undecalcified sections were examined using transmitted and ultraviolet light microscopy. Mechanical testing showed one HA formulation provided 70-100% bonding. Histology showed intimate contact between the gage and bone surface. Histomorphometry indicated increased bone activity under the gage compared to the remaining bone, the controls, and the shams. The results indicate that microcrystalline HAs bond to bone quickly and can allow long term in vivo measurements.
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Procion brilliant red H-8 BS is a fluorescent dye that stains the organic matrix of bone supravitally. A procedure is described for counterstaining sections of such bone for visible light examination without interfering with the demonstration of sites of bound dye under UV illumination. Sections are brought to water, stained in Delafield's alum hematoxylin for 10 minutes, washed in tap water for 10 minutes, counterstained in 1% Procion brillant orange M-GS for 15 minutes and washed in distilled water for 10 minutes. After dehydration the sections were mounted in Eukitt.
About 3.5 cm osteoperiosteal circumferential gaps in rabbit ulnae were bridged by partially decalcified (by 0.6 M HCl) allogenic implants, 8-12 weeks after implantation complete bridging of the gap by new bone formation and union at host-graft junctions was observed in 97.2 per cent of the experiments. Instant permeability and the powerful osteo-inductive potential of the implant coupled with some degree of mechanical stability provided by partial decalcification ensured successful bone formation in almost all the experiments. It is suggested that partially decalcified allogenic bone matrix preserved in 70 per cent ethanol may be a reasonably good substitute for autologous bone graft.
OBJECT: Convection-enhanced delivery (CED) can be used safely to achieve high local infusate concentrations within the brain and spinal cord. The use of CED in the brainstem has not been previously reported and may offer an alternative method for treating diffuse pontine gliomas. In the present study the authors tested CED within the rat brainstem to assess its safety and establish distribution parameters. METHODS: Eighteen rats underwent stereotactic cannula placement into the pontine nucleus oralis without subsequent infusions. Twenty rats underwent stereotactic cannula placement followed by infusion of fluorescein isothiocyanate (FITC)-dextran at a constant rate (0.1 microl/minute) until various total volumes of infusion (V(i)s) were reached: 0.5, 1, 2, and 4 microl. Additional rats underwent FITC-dextran infusion (V, 4 microl) and were observed for 48 hours (five animals) or 14 days (five animals). Serial (20-microm thick) brain sections were imaged using confocal microscopy with ultraviolet illumination, and the volume of distribution (Vd) was calculated using computer image analysis. Histological analysis was performed on adjacent sections. No animal exhibited a postoperative neurological deficit, and there was no histological evidence of tissue disruption. The Vd increased linearly (range 15.4-55.8 mm3) along with increasing Vi, with statistically significant correlations for all groups that were compared (p < 0.022). The Va/Vi ratio ranged from 14 to 30.9. The maximum cross-sectional area of fluorescence (range 9.8-20.9 mm2) and the craniocaudal extent of fluorescence (range 2.8-5.1 mm) increased with increasing Vi. CONCLUSIONS: Convection-enhanced delivery can be safely applied to the rat brainstem with substantial and predictable V(d)s. This study provides the basis for investigating delivery of various candidate agents for the treatment of diffuse pontine gliomas.
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