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At least 1,081 records · Page 60Linked to original sources
PubMed1981

Characterization of a variety of standard collagen substrates: ultrastructure, uniformity, and capacity to bind and promote growth of neurons.

Collagen substrates were characterized after preparation by the four methods most commonly used for tissue culture (saline precipitation, exposure to ammonium hydroxide vapor, exposure to ultraviolet light, and air drying). Although roughly equivalent percentages of collagen were precipitated by each technique (87 to 97%), marked differences were found in surface uniformity and ultrastructure. Substrates were quite uniform if precipitated by exposure to ammonium hydroxide or ultraviolet light, of intermediate uniformity if saline precipitated, and not at all uniform if air dried. Scanning electron microscopy revealed that (a) ammonium hydroxide and saline precipitation primarily resulted in formation of collagen fibrils, (b) air drying produced a small number of fibrils plus a large amount of amorphous material, and (c) exposure to ultraviolet light only resulted in the formation of globular, nonfibrillar collagen aggregates. The capacity of collagen substrates to bind and grow neurons differed markedly with the method of preparation and the amount of collagen plated per unit area. Quantifications of binding and growth of both cerebral and sympathetic neurons revealed that these are separate measures of the biocompatibility of a surface and that growth was uniformly inferior on globular collagen that had been precipitated by ultraviolet light. Long-term (greater than or equal to 2 wk) growth of sympathetic neurons was optimal on thick beds of saline-precipitated collagen, whereas short-term growth was best on thin layers of either saline or ammonium hydroxide-precipitated collagen. Cerebral neurons bound and grew optimally on thick collagen beds after both short- and long-term culture. In addition, cerebral neurons were found to be more dependent on the method of precipitation of the thin collagen substrates than were sympathetic neurons.

Ammonium Hydroxide↗
PubMed1986

Light-polymerizing plastics as slide mounting media.

Currently available mounting media require solvent evaporation for hardening. This process is slow and often is complicated by the formation of air spaces under the coverslip. Light-polymerizing plastics are introduced as histologic mounting media that eliminate these problems because they harden quickly when exposed to ultraviolet light and, therefore, do not depend upon solvent evaporation. These light-polymerizing plastics are easy to use, permit rapid and controlled hardening, and eliminate the formation of air spaces under the coverslip. The optical characteristics are satisfactory for light and fluorescence microscopy.

Animals↗
PubMed1975

[Herpes Simplex Keratitis under the Scanning lectron Microscope(author's transl)].

In 12 rabbits a dendritic keratitis could be provoked by applying a suspension of herpes viruses (Herpes virus hominis -- HVH type 1) onto their previously both uviolised and non-uviolised corneae. 4 days later ophthalmectomy was performed and the corneae were examined by the scanning electron microscope. Macroscopically, both in the uviolised eyes and the control eyes punctiform lesions of the corneae were observed. After short ray-treatment (3 min) no differences were present between the uviolised and non-uviolised eyes. At longer time of exposure (7--15 min), however, a more distinct alteration occurred in the uviolised eye. The scanning-electron-microscopical picture showed the dendritic lesions to be composed of numerous circular foci of 100 to 200 mu. There were no differences in the lesions of the uviolised and non-uviolised corneae. The fact that after ultraviolet ray-treatment a great part of the epithelial cells is sloughed off, but, nevertheless, the same dendritic lesions develop, contradicts the assumption of a morphological substrate of the epithelial cells being the inducing factor.

Animals↗
PubMed1967

Tyrosinase inhibition: its role in suntanning and in albinism.

Tyrosinase inhibitor (molecular weight less than 5000; extracted from various melanomas) fully inhibits soluble tyrosinase but only partially inhibits tyrosinase "aggregated" into melanosomes; the inhibitor can be inactivated by ultraviolet light. S91 Albinotyrosinase Type B apparently cannot "aggregate" into melanosomes because its protein carrier is genetically altered. Therefore, albinotyrosinase remains vulnerable to its inhibitor and cannot produce melanin, even though the enzyme has a functioning active center.

Albinism↗
PubMed1988

UV-induced growth inhibition in the trypanosomatid Crithidia fasciculata.

The effects of short ultraviolet (253 nm) radiation on the growth of the trypanosomatid flagellate Crithidia fasciculata were studied in liquid medium and nutrient agar plates. Cell duplication was completely inhibited after exposure of the flagellates to doses equal to or higher than 50 J/m2. The UV-induced lag period was dose-dependent. Survival was reduced to 1% after exposure of the parasites to 200 J/m2. Ultrastructural changes after the lag period were studied by transmission electron microscopy. Changes of the kinetoplast network structure and sometimes of the mitochondrial matrix were observed. The existence of DNA repair mechanisms in this protozoan is discussed.

Animals↗
PubMed1989

Asymmetric behavior of severed microtubule ends after ultraviolet-microbeam irradiation of individual microtubules in vitro.

The molecular basis of microtubule dynamic instability is controversial, but is thought to be related to a "GTP cap." A key prediction of the GTP cap model is that the proposed labile GDP-tubulin core will rapidly dissociate if the GTP-tubulin cap is lost. We have tested this prediction by using a UV microbeam to cut the ends from elongating microtubules. Phosphocellulose-purified tubulin was assembled onto the plus and minus ends of sea urchin flagellar axoneme fragments at 21-22 degrees C. The assembly dynamics of individual microtubules were recorded in real time using video microscopy. When the tip of an elongating plus end microtubule was cut off, the severed plus end microtubule always rapidly shortened back to the axoneme at the normal plus end rate. However, when the distal tip of an elongating minus end microtubule was cut off, no rapid shortening occurred. Instead, the severed minus end resumed elongation at the normal minus end rate. Our results show that some form of "stabilizing cap," possibly a GTP cap, governs the transition (catastrophe) from elongation to rapid shortening at the plus end. At the minus end, a simple GTP cap is not sufficient to explain the observed behavior unless UV induces immediate recapping of minus, but not plus, ends. Another possibility is that a second step, perhaps a structural transformation, is required in addition to GTP cap loss for rapid shortening to occur. This transformation would be favored at plus, but not minus ends, to account for the asymmetric behavior of the ends.

Guanosine Diphosphate↗
PubMed2005

[Ultraviolet and blue-violet photoluminescence of gold nanoparticles].

Suspended gold nanoparticles (size range 20-30 nm) have been synthesized via electrochemical method. The emission spectra of gold colloidal nanoparticles were studied at room temperature. Fluorescence occurs at ultraviolet and blue-violet wavelengths. Two emission peaks were observed at 377 nm and 459 nm, respectively, when the corresponding excitation wavelength was at 220 nm. The emission peak at 377 nm increases with increasing particle density or excitation intensity, whereas the emission peak at 459 nm decreases with decreasing excitation intensity or increasing the particle density, and disappears below the threshold. With increasing the slit width, the intensity difference between these two emission peaks decreases and the ratio approaches 1. All these observed results are in agreement with the theory of self-organized random micro-cavity. It is indicated that the fluorescence emissions in ultraviolet and blue-violet wavelength regions are induced by the multiple scattering in a disordered gold nanoparticles system. And these photoluminescence features suggest the possible future applications in the areas of optical data storage and full color display.

Gold↗
PubMed1995

Osmium tetroxide-potassium ferrocyanide intensification of a diaminobenzidine product obtained by photoconversion of a fluorescent label: a study of human neutrophil granules.

A diaminobenzidine posttreatment employing osmium tetroxide and potassium ferrocyanide was successfully used to intensify the diaminobenzidine stain formed by photoconversion of immunofluorescent labelling. Lactoferrin labelled granules became visible following the photoconversion process. Adequate diaminobenzidine staining was obtained after irradiating the cytospin preparations with ultraviolet light for 30-40 min. The diaminobenzidine stain had advantages over the fluorescent stain owing to its greater stability, greater density, and ability to be intensified. The enhancement procedure intensified both the color and density of the diaminobenzidine product. Consequently, shorter irradiation times could be used. Osmium tetroxide solutions of 3-4% increased the intensity with minimal background staining. Ultrastructural immunogold cytochemistry on ultrathin sections confirmed the existence of the lactoferrin labelled structures observed by light microscopy of cytospin preparations indicating that these were secondary granules. The photomicroscopy procedures used in this study were simple to perform and could be applied to studies of other cellular antigens prior to using immunoultramicroscopy.

3,3'-Diaminobenzidine↗
PubMed1995

Retrograde but not anterograde bead movement in intact axons requires dynein.

Dynein and kinesin have been implicated as the molecular motors that are responsible for the fast transport of axonal membranous organelles and vesicles. Experiments performed in vitro with partially reconstituted preparations have led to the hypothesis that kinesin moves organelles in the anterograde direction and dynein moves them in the retrograde direction. However, the molecular basis of transport directionality remains unclear. In the experiments described here, carboxylated fluorescent beads were injected into living Mauthner axons of lamprey and the beads were observed to move in both the anterograde and retrograde directions. The bead movement in both directions required intact microtubules, occurred at velocities approaching organelle fast transport in vivo, and was inhibited by vanadate at concentrations that inhibit organelle fast transport. When living axons were injected with micromolar concentrations of vanadate and irradiated at 365 nm prior to bead injections, a treatment that results in the V1 photolysis of dynein, the retrograde movement of the beads was specifically abolished. Neither the ultraviolet irradiation alone nor the vanadate alone produced the retrograde-specific inhibition. These results support the hypothesis that dynein is required for retrograde, but not anterograde, transport in vivo.

Animals↗
PubMed1998

Variable expression of apoptotic phenotype in keratinocytes treated with ultraviolet radiation, ceramide, or suspended in semisolid methylcellulose.

Apoptosis is a form of cellular suicide and is activated in various cells, including keratinocytes, in response to physiological and pathological stimuli. A current hypothesis holds that apoptotic cells in a concerted manner express characteristic phenotypic features comprising cytoplasmic budding, pyknosis, chromatin condensation, karyorhexis and internucleosomal DNA fragmentation. In this study we investigated the effects of different potential inducers of apoptosis on cultured human keratinocytes. Viability was determined with vital dyes (ethidium bromide, trypan blue), cell morphology was investigated with electron microscopy, and nuclear and DNA integrity was assessed with TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling) or DNA gel electrophoresis. Irradiation with 50 mJ/cm2 ultraviolet B (UVB), treatment with C8 ceramide, or suspension in a semisolid medium caused apoptosis which was ultrastructurally different from necrotic induced by very high (300 mJ/cm2) doses of UVB. However, the phenotype of dying cells did not exhibit all typical features of apoptosis and cell morphology depended on the method used to induce apoptosis. Cells irradiated with ultraviolet B or treated with C8 ceramide developed large and small budding and DNA nicks but not chromatin condensation or classical karyorhexis. In UVB-irradiated cells a novel form of karyorhexis was observed manifested by formation of a few very small chromatin fragments in nuclear periphery. Cells suspended in methylcellulose developed DNA nicks and pyknosis, but not budding or karyorhexis. In neither case could the typical internucleosomal DNA fragments be detected by gel electrophoresis. These morphological results indicate that in keratinocytes induction of effect or death mechanisms is not concerted but depends on the stimulus inducing apoptosis.

Apoptosis↗
PubMed1992

Techniques for converting Golgi precipitate in CNS neurons into stable electron microscopic markers.

Direct electron microscopy of nervous tissue stained with the Golgi impregnation method is unsatisfactory because the cytoplasm of the cell bodies and processes of the impregnated neurons are completely filled with a compact precipitate of electron dense silver chromate. This precipitate entirely obscures the cytological details of the impregnated neurons. Because of its solidity and instability in aqueous solutions, the silver chromate is also a source of inconvenience during the preparation of the ultrathin sections. This review summarizes methods that have been developed with the aim of replacing the Golgi precipitate in CNS neurons with a more convenient electron dense material--for example, heavy metal salts or metallic particles. Conversion of the precipitate into a stable electron dense marker is done before the material is embedded for electron microscopy. The methods include lead, gold, and bromide substitution, treatment with ammonia, direct chemical reduction into metallic silver, and photoreduction of the silver chromate into silver through irradiation with ultraviolet light.

Aminophenols↗
PubMed2000

Two-photon excitation of fluorescence for three-dimensional optical imaging of biological structures.

Techniques based on two-photon excitation (TPE) allow three-dimensional (3D) imaging in highly localized volumes, of the order of magnitude of a fraction of a femtolitre up to single-molecule detection. In TPE microscopy a fundamental advantage over conventional widefield or confocal 3D fluorescence microscopy is given by the use of infrared (IR) instead of ultraviolet (UV) radiation to excite those fluorophores requiring UV excitation, hence causing little damage to the specimen or to fluorescent molecules outside the volume of the TPE event and allowing a deeper penetration within the sample compared with conventional one-photon excitation of fluorescence. In our laboratory, within the framework of a national INFM project, we have realized a TPE fluorescence microscope, part of a multipurpose architecture also including lifetime imaging and fluorescence correlation spectroscopy modules. The core of the architecture is a mode-locked Ti:sapphire infrared pulsed laser pumped by a high-power (5 W, 532 nm) solid-state laser and coupled to an ultracompact scanning head. For the source we have measured a pulse width from 65 to 95 fs as a function of wavelength (690-830 nm). The scanning head allows conventional and two-photon confocal imaging. Point spread function measurements are reported with examples of applications to the study of biological systems.

Animals↗
PubMed2005

Inactivation of particle-associated viral surrogates by ultraviolet light.

This study investigated whether colloid-sized particles can enmesh and protect viruses from 254-nm ultraviolet (UV) light and sought to determine the particle characteristics (e.g. size, chemical composition) that are most relevant in causing a protective effect. Two viral surrogates (MS2 coliphage and bacteriophage T4), three types of particles (kaolin clay, humic acid powder, and activated sludge), two coagulants (alum and ferric chloride), two filtration conditions (none and 0.45 microm), and two UV doses (40 and 80 mJ/cm2 for MS2 coliphage; 2 and 7 mJ/cm2 for bacteriophage T4) were considered in a series of bench-scale UV collimated beam experiments. Transmission electron microscopy was used to qualitatively confirm the phage particle-association after coagulation. Humic acid and activated sludge floc particles shielded both viral surrogates to a statistically significant degree (with >99% confidence) relative to particle-free control conditions, while the kaolin clay particles provided no significant protection. The results of the study suggest that particles <2 microm in diameter are large enough to protect viruses from UV light and that particulate chemical composition (e.g. UV-absorbing organic content) may be a critical factor in the survival of particle-associated viruses during UV disinfection.

Alum Compounds↗