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PubMed1980

The use of fluorescein diacetate to assess embryo viability in the mouse.

Preimplantation mouse embryos that were exposed to fluorescein diacetate (FDA) accumulated intracellular fluorescein and fluoresced brightly under ultraviolet (u.v.) light. The rate at which intracellular fluorescein was lost from the cells was measured at 37, 28 and 4 degrees C and the rate decreased as the storage temperature decreased. The rate at which intracellular fluorescein accumulated increased as FDA concentration increased until a maximum rate was attained. The ability to accumulate intracellular fluorescein could be removed by heating embryos at 56 degrees C for 30 min or by damaging the cell membrane. Cells grown under inadequate culture conditions lost the ability to accumulate intracellular fluorescein. Exposure of 2-cell mouse embryos to FDA and u.v. light did not alter the rate of blastocyst formation in vitro, and exposure of blastocysts to FDA and u.v. light did not alter the rate of implantation or post-implantation development in vivo.

Animals↗
PubMed1992

Characterization of ultraviolet laser-induced autofluorescence of ceroid deposits and other structures in atherosclerotic plaques as a potential diagnostic for laser angiosurgery.

Unstained frozen sections of normal and atherosclerotic human aorta and coronary artery were examined using histochemical and fluorescence microscopic techniques to identify the structures responsible for autofluorescence under 351 to 364 nm laser excitation. These structures included elastin and collagen in normal and atherosclerotic specimens, calcium deposits in calcified plaques, and granular or ring-shaped deposits histochemically identified as ceroid found in both calcified and non-calcified plaques. Qualitatively, both the color and intensity of ceroid autofluorescence differed greatly from that of elastin or collagen. The emission spectra of elastin, collagen, and ceroid were examined by microscopic spectrofluorimetry, and were found to differ significantly as well. When compared with spectra of elastin and collagen, spectra of ceroid were broader, shifted to the red, and were somewhat resistant to bleaching. We conclude that detection of laser-induced ceroid autofluorescence may aid in identifying plaques for laser ablation.

Angioplasty, Laser↗
PubMed2004

Study of the incidence and nature of "very subtle epidermal melasma" in relation to intense pulsed light treatment.

BACKGROUND: Skin rejuvenation with intense pulsed light (IPL) is effective for clearing epidermal pigment disorders. Complications are mild and limited to epidermal burns caused by excessive settings. Some patients, however, experience IPL-induced melasma-like hyperpigmentation despite the appearance of normal skin. These patients seem to have very subtle epidermal melasma not visible to the naked eye. Ultraviolet photography has been useful in identifying these patients and preventing complications. OBJECTIVE: The study investigated the incidence of very subtle melasma in patients using UV photography, and assessed this tool in identifying high-risk patients. SUBJECTS AND METHODS: 223 Japanese women, 30-69 years old, participated in the study. Very subtle melasma invisible to the naked eye under normal light was diagnosed by UV photography by two physicians, and any relationship among the disease incidence, age, and regular sunscreen use was examined. RESULTS: Sixty-three cases of very subtle melasma (28.3%) were identified among the 223 subjects, with a significantly lower incidence in sunscreen users. CONCLUSIONS: Patients diagnosed with subtle epidermal melasma and treated with mild IPL parameters did not suffer induced secondary hyperpigmentaion. To help avoid complications after treatment, IPL users should be aware of the age and sunscreen-related incidence of this phenomenon in Asian patients.

Adult↗
PubMed1990

Acridine orange fluorescence, Campylobacter pylori, and chronic gastritis.

Endoscopic gastric biopsy specimens from 230 consecutive patients in a North Liverpool District were histologically studied by routine light microscopy and ultraviolet fluorescence after acridine orange staining. Eighty patients with chronic gastritis were further studied with regard to type of gastritis and its activity, presence of Campylobacter pylori (CP), and degree of colonization of the gastric mucosa. Miscellaneous gastritis, gastric ulcers, erosions, neoplasms, and histologically normal specimens were excluded from the study. The results show statistically significant correlation between chronic gastritis and CP (P = 0.01, Mann-Whitney test). The activity of gastritis correlated well with CP, but there was no statistical significance between the density of neutrophils and degree of CP colonization (P greater than 0.5, Mann-Whitney test). In a small sample of CP-positive specimens acridine orange stain was compared with Warthin-Starry, Giemsa (modified), cresyl fast violet, and haematoxylin and eosin stains. Acridine orange stain in the histologic identification of CP has been used only once before in a large study.

Acridine Orange↗
PubMed1996

Biomedical three-dimensional holographic microimaging at visible, ultraviolet and X-ray wavelengths.

A new imaging technology is under advanced development that has several key advantages over conventional forms of microimaging performed with standard light microscopes, confocal light microscopes, and electron microscopes. The image created by this microscope possesses several unique features: It is intrinsically three-dimensional; it can be formed with very high contrast, a characteristic of the phase-sensitive nature of the recording; the information contained in the image is obtained in a single exposure of the specimen, a feature that eliminates the accumulation of damage to living systems that can occur with techniques utilizing multiple exposures; the method of image construction is fundamentally free from aberration (distortion), thereby obviating the need to employ complex procedures for correction; the exact focal plane of any optical section is digitally determined through computation and is not based on any mechanical adjustments; and the principles of operation, including the computational processes and modalities of image presentation, are uniform over the full range of spectral coverage spanning from the visible (approximately 500 nm) to the X-ray (approximately 0.3 nm) regions. The application of this new technology is expected to open new cost-effective avenues to the understanding, prevention and treatment of broad areas of human disease.

Animals↗
PubMed2006

Sequential estimation of optical properties of a two-layered epithelial tissue model from depth-resolved ultraviolet-visible diffuse reflectance spectra.

A method for estimating the optical properties of two-layered media (such as squamous epithelial tissue) over a range of wavelengths in the ultraviolet-visible spectrum is proposed and tested with Monte Carlo modeling. The method first used a fiber-optic probe with angled illumination and the collection fibers placed at a small separation ( or=1000 microm) was used to detect diffuse reflectance preferentially from the bottom layer. A second Monte Carlo-based inverse model for a two-layered medium was applied to estimate the bottom layer optical properties, as well as the top layer thickness, given that the top layer optical properties have been estimated. The results of Monte Carlo validation show that this method works well for an epithelial tissue model with a top layer thickness ranging from 200 to 500 microm. For most thicknesses within this range, the absorption coefficients were estimated to within 15% of the true values, the reduced scattering coefficients were estimated to within 20% and the top layer thicknesses were estimated to within 20%. The application of a variance reduction technique to the Monte Carlo modeling proved to be effective in improving the accuracy with which the optical properties are estimated.

Algorithms↗
PubMed1972

Some quantitative observations upon the responses of neuroglial cells which follow axotomy of adjacent neurones.

1. The dry mass and nucleic acid content of freshly isolated neuroglial cells and of their nucleoli were measured by interference microscopy and ultraviolet absorption microspectrography. The incorporation of tritiated nucleosides and of an amino acid was followed autoradiographically.2. After hypoglossal axotomy in the adult rat hypertrophy of astrocytes of the hypoglossal nucleus occurred in a biphasic manner. The first phase lasted from days 1-10 and was accompanied by a small degree of astrocytic hyperplasia, and the second from days 20-80. Hypertrophy of oligodendrocytes accompanied the second phase of the astrocytic response.3. When severed axons failed to reinnervate denervated muscle, the second phase of the astrocytic response was markedly reduced and the hypertrophy of oligodendrocytes did not occur.4. If the severed axons re-innervated denervated muscle after a controlled delay, the second phase of the astrocytic response and the oligodendroglial hypertrophy was also delayed.5. Injection of botulinum toxin into the tongue caused changes in astrocytes and oligondendrocytes closely resembling those found after axotomy.6. Transient astrocytic hypertrophy occurred in the uninjured right hypoglossal nucleus, and had a different time course to the changes occurring on the injured side.7. The results are discussed in relation to changes in the metabolism and to alterations in the dendritic fields of injured neurones, previously measured in these circumstances.

Animals↗
PubMed2002

Metabolic pathways of carcinogenic chromium.

The products of hexavalent chromium [Cr(VI)] reduction by glutathione (GSH) alone or in the presence of equimolar quantities of aspartate (Asp) and/or glutamate (Glu) and a chromium-containing material extracted from bovine liver were studied by ultraviolet-visible spectrum (UV-vis) studies, electrospray mass spectrometry (ES-MS), electron paramagnetic resonance (EPR), and nuclear magnetic resonance (NMR). Reduction of chromate by GSH was followed by UV-vis and NMR, revealing the formation of a paramagnetic complex in which GSH acts as a ligand. ES-MS and EPR measurements provided unequivocal evidence of a dimeric Cr(V)(2)GSH(2) species in which the two metal ions are bridged by the Gamma-Glu carboxylate. The analysis of the (1)H and (13)C shifts experienced by GSH protons and the values of paramagnetic contributions to proton spin-lattice relaxation rates provided a set of constraints for structural determination. The same experiments were repeated in the presence of an equimolar concentration of Asp, revealing the formation of a dimeric Cr(V) paramagnetic complex in which the two metals are now bridged by Asp. Nuclear magnetic resonance dispersion profiles show that water is not displaced by Asp and that the correlation time of this complex is slowed by the increased complexity. When Glu is also included in the solution in equimolar concentration to GSH and Asp, data are consistent with the formation of many mono- and dinuclear species, with the three ligands competing with each other. Finally, the spectroscopic investigation of the chromium-containing material extracted from bovine liver revealed the presence of a complicate mixture of Cr(IV) or Cr(V) complexes, among which some Cr(V)-GSH species are present alone or with other ligands in the metal coordination sphere.

Aspartic Acid↗
PubMed1996

Hepatocyte death following transforming growth factor-beta 1 addition.

Apoptosis is a morphological term which describes a sequence of events finally leading to cell death. In epithelial organs, induction of cell death is closely linked to an inhibitor of epithelial growth, transforming growth factor-beta 1 (TGF-beta 1). In this paper, we describe the morphology of TGF-beta 1-induced apoptosis in hepatocytes of the hyperplastic liver and primary cultures. Chromatin condensation, a hallmark of apoptosis, was observed in primary hepatocytes by confocal and vital UV microscopy. In addition, we have applied the morphological detection of DNA strand breaks both by in situ tailing (ISTAIL) and in situ nick translation (ISNT).

Animals↗
PubMed1996

Laboratory implementation of a rapid three-stain technique for detection of microorganisms from lower respiratory specimens.

A rapid, cost-effective method for the evaluation of lower respiratory specimen has become increasingly important in the diagnosis of pulmonary diseases in immunocompromised patients. In the past, the technically demanding, time-consuming, and expensive Gomori-methenamine-silver (GMS) stain was the principal means for the evaluation of these specimens. In this study, we compared the GMS stain with a new rapid, three-stain protocol for the evaluation of lower respiratory specimens. Lower respiratory specimens were obtained by bronchoalveolar lavage (BAL). Conventional Wright/Giemsa and Gram stains were utilized, as well as a contemporary strain, calcofluor white (CW). A cell count was performed on the BAL specimens, and cytospins were stained by the three stains. The calcofluor white-stained slides were examined with an epi-fluorescent microscope, whereas the other stains were evaluated with a conventional light microscope. Gomorimethenamine-silver (GMS), acid-fast bacillus (AFB), and Papanicolaou (PAP) stains were performed as controls. Thirty-two BAL procedures were performed in 20 (63%) male patients and 12 (37%) female patients. The clinical diagnosis was pneumonia in 31% of the patients, malignant hematologic disease in 28%, acute respiratory distress syndrome (ARDS) in 9%, and acquired immunodeficiency syndrome (AIDS) in 28%. Of these specimens, 78% were adequate for interpretation and 22% were inadequate. Bacteria were found in 50% (16/32) of all BALs, fungi were found in 9% (3/32), and Pneumocystis carinii was found in 9% (3/32). Gram-positive bacteria were most frequently found in patients with pneumonia (80%, 4/5), whereas P. carinii was identified in patients with AIDS. There were no false-positive results. One CW stain was equivocal for P. carinii due to high fluorescent background. Laboratory implementation of the rapid, three-staining technique was accomplished without difficulty in microbiology and hematology laboratory sections. Specimen evaluation with the rapid staining protocol was technically easy to perform; however, experience in ultraviolet fluorescent microscopy was crucial for interpretation of CW stain. All results were available in 2 hr, cost was reduced by 30%, and the assays were available 7 days/week. Further studies are ongoing to substantiate the sensitivity, specificity, and predictive value of this technique, as well as clinical guidelines for its optimal utilization.

Adult↗
PubMed2007

Determination of polymethoxylated flavones in peels of selected Jamaican and Mexican citrus (Citrus spp.) cultivars by high-performance liquid chromatography.

The concentrations of the polymethoxylated flavones (PMFs) in peels of selected citrus cultivars grown in Jamaica and Mexico were determined. The PMFs were extracted from sun-dried citrus peels with reagent-grade methanol. Analyses were carried out by reverse-phase HPLC and UV detection. The column used was a C(18) 5 microm (150 x 4.6 mm) Discovery column. Elution was in the gradient mode, using a ternary mobile phase. The results showed that all the citrus cultivars used contained at least three of the six major PMFs quantified. Ortanique peel contained the highest quantity of PMFs (34,393 +/- 272 ppm), followed by tangerine (28,389 +/- 343 ppm) and Mexican sweet orange (sample 1; 21,627 +/- 494 ppm). The major PMFs, i.e. sinensetin, nobiletin, tangeretin, heptamethoxyflavone, tetramethylscutellarein and hexamethyl-o-quercetagetin, present in the peels of 20 citrus cultivars, was quantified. The results were compared with those of Florida citrus peels. A large amount of citrus peels and byproducts are produced in the Caribbean which could provide a cheap and convenient source of PMFs.

Calibration↗
PubMed1981

Cytophotometric analysis of lytically and persistently infected tissue culture cells with measles virus.

Lytically and persistently infected VERO and LUB cells were cytophotometrically characterized with the aid of computer analysis. Images scanned at 260 and 280 nm were processed by computer algorithms. With this approach, infected cells could be segmented and differentiated from uninfected cells. Lytically and persistently infected cells could be distinguished by distinct differences in nuclear and cytoplasmic optical densities. These findings are supported by biologic data based on the analysis of virus-specific proteins and nucleic acids. The applied computer aided cytophotometry provides a new approach in the study of virus cell interaction.

Animals↗
PubMed1992

Preparation and microscopic visualization of multicolor luminescent immunophosphors.

The preparation of charge-stabilized suspensions of small phosphor particles (0.1-0.3 micron) and their coupling with antibodies to immunoreactive conjugates is described. Phosphor particles consisting of yttriumoxisulfide activated with europium served as a model system in the evaluation of the stabilizing properties of several polycarboxylic acids. The optimal reagents were then applied to other phosphors which differ in spectral characteristics as well as in luminescence lifetime. These phosphors were ground to a size of 0.1-0.3 micron and proteins or other macromolecules were adsorbed to the phosphor particles to prepare conjugates of different physico-chemical properties. A time-resolved microscope, suitable for real time visualization of the time-delayed luminescence of the immunophosphors by the human eye, is described in detail. Since most phosphors require excitation with far UV light, a special fluorescence microscope allowing far UV excitation was developed for conventional visualization of the luminescence emitted by the phosphor. The possibility of multiple color labeling using various phosphor conjugates was demonstrated in a model system consisting of haptenized latex beads.

Adsorption↗
PubMed1990

Class I and class II major histocompatibility complex antigen expression on hepatocytes: a study in children with liver disease.

Controversy exists regarding major histocompatibility complex antigen expression on hepatocytes. In this study, hepatocyte expression of class I and II major histocompatibility complex antigens was investigated in diseased and normal livers, using indirect immunofluorescent staining of mechanically isolated, viable hepatocytes. Hepatocytes were obtained from 76 children: 10 with autoimmune chronic active hepatitis, nine with primary sclerosing cholangitis, nine with chronic hepatitis B virus infection, five after liver transplantation, 19 with extrahepatic biliary atresia, 11 with alpha 1-antitrypsin deficiency, four with idiopathic neonatal hepatitis and nine with histologically normal liver. Immunohistochemistry was performed in all cases; flow cytofluorimetry was performed for class I antigens in 38 cases and performed for class II antigens in 18 cases. From three children with autoimmune chronic active hepatitis and two with chronic hepatitis B virus infection, isolated hepatocytes were also incubated with gamma-interferon before staining and analysis. By fluorescence microscopy, class I antigens were detected on hepatocytes from all children, the highest percentage (100%) of positive cells and the most intense staining were observed in untreated patients with autoimmune chronic active hepatitis or primary sclerosing cholangitis and in those with acute rejection of a liver transplant. Reduced class I antigen expression occurred in chronic hepatitis B virus infection. Class II antigens were only detected on hepatocytes from eight patients: three with autoimmune chronic active hepatitis and five with primary sclerosing cholangitis, all untreated. Flow cytofluorimetric analysis confirmed the results obtained by fluorescence microscopy, but it also demonstrated a weak class II antigen expression during liver allograft rejection.(ABSTRACT TRUNCATED AT 250 WORDS)

Biopsy↗
PubMedDate not supplied

Ultrastructural examination of rabbit aortic wall following high-fat diet feeding and selenium supplementation: a transmission electron microscopy study.

Experiments were carried out to examine the changes occurring in the wall of rabbit aortae following high-fat diet (HFD) feeding as well as HFD + selenium supplementation. Male New Zealand White rabbits were divided into three groups-control, HFD-fed and HFD + Se supplementation-and were treated for three months. The study depicted that levels of serum total cholesterol and triglycerides were markedly increased in the HFD-fed group as compared with control animals. However, in the HFD + Se-fed group, these levels were markedly suppressed vis-à-vis animals fed on HFD only. Development of atherogenic and atheromatic plaques has been shown at the light microscopy level in HFD-fed rabbits, whereas these developments were not visible in the HFD + Se-fed rabbits. Transmission electron microscopy findings indicated altered ultrastructure in the endothelial cells of the intimal layer as well as smooth-muscle cells of the medial layer in HFD-fed animals. However, these findings indicated normal ultrastructure in most of the cells, with little ultrastructural alterations from animals supplemented with Se along with HFD feeding. The study on the whole depicted the ability of Se to inhibit the onset of progression of aortic disease and hence has relevance to its therapeutic potential.

Animals↗
PubMed1989

Evaluation of axonal specificity of the regenerating rat sciatic nerve using a fluorescent labeling technique.

A double-labeling technique using fluorescent compounds, i.e., diamidino yellow and true blue, is described, with the divided rat sciatic nerve model. Tibial and peroneal fascicles were transected, and each freshly cut nerve end was placed in a vial containing either the true blue or the diamidino yellow label. After a 10-15 day survival period, animals were systemically perfused with fixative, and the lumbar enlargement and dorsal root ganglia corresponding to sciatic nerve roots were removed. Serial sectioning and examination with ultraviolet fluorescence microscopy and counts of the labeled cells were performed. Diamidino yellow in labeled cells was found localized to the nucleus, whereas true blue was found localized to the cytoplasm and nucleolus (control group n = 5). Intermixing of these pools was noted, with yellow cells seen among blue labeled cells following nerve autograft repair of a 1 cm segment of the rat sciatic nerve (experimental group n = 5). Double-labeled cells displaying a yellow nucleus and a blue cytoplasmic label were also seen. We believe that the double-labeling technique described, using the fluorescent retrograde tracers diamidino yellow and true blue, is a useful monitoring method for assessing the specificity of regenerating axons in the rat sciatic nerve.

Amidines↗