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At least 901 records · Page 50Linked to original sources
PubMed2002

Hypoglycaemic effect of a novel insulin buccal formulation on rabbits.

Transmucosal delivery is a suitable route for insulin non-injection administration. In this study, the hypoglycaemic effect of INSULIN BUCCAL SPRAY (IBS), a formulation with soybean lecithin and propanediol combined as absorption enhancer for insulin on diabetic rabbits and rats, were investigated. The hypoglycaemic rate was calculated and the pharmacodynamics and pharmacokinetics of the formulation in rabbits were studied. The results show that when the diabetic rabbits were administrated with IBS in dosages of 0.5, 1.5 and 4.5Ukg(-1), the blood glucose level decreased significantly compared with that of the control group and the hypoglycaemic effect lasted over 5h. The blood glucose decreasing rates are 22.4, 48.1 and 53.5%, respectively. The average bioavailability of IBS by buccal delivery versus subcutaneous injection is 29.2%. Meanwhile, the diabetic rats were administrated with IBS in dosages of 1.0, 3.0 and 9.0Ukg(-1), the blood glucose level decreased significantly compared with that of the control group and the hypoglycaemic effect lasted over 4h. The blood glucose decreasing rates are 24.6, 47.5 and 59.6%, respectively. Furthermore, the penetration of fluorescein isothiocyanate (FITC)-labelled insulin through rabbit buccal mucosa was investigated by scanning the distribution of the fluorescent probe in the epithelium using confocal laser scanning microscopy. The results revealed that FITC-insulin can pass through the buccal mucosa promoted by the enhancer and the passage of insulin across the epithelium includes both intracellular and paracellular routes. From the rabbit and rat experimental results showed that IBS is an effective buccal delivery system, which is promising for clinical trial and the future clinical application.

Administration, Buccal↗
PubMed2006

Cell mechanics using atomic force microscopy-based single-cell compression.

We report herein the establishment of a single-cell compression method based on force measurements in atomic force microscopy (AFM). The high-resolution bright-field or confocal laser scanning microscopy guides the location of the AFM probe and then monitors the deformation of cell shape, while microsphere-modified AFM probes compress the cell and measure the force. Force and deformation profiles of living cells reveal a cubic relationship at small deformation (<30%), multiple peaks at 30-70% compression, and a rapid increase at over 80% deformation. The initial compression may be described qualitatively and quantitatively using a simple model of a nonpermeable balloon filled with incompressible fluid. Stress peaks reflect cell membrane rupture, followed by the deformation and rupture of intracellular components, beyond which the cell responses become irreversible. The Young's modulus and bending constant of living cell membranes are extracted from the balloon models, with 10-30 MPa and 17-52 kT, respectively. The initial compression of dead and fixed cells is modeled using Hertzian contact theory, assuming that the cell is a homogeneous sphere. Dead cells exhibit a cytoskeleton elasticity of 4-7.5 kPa, while fixation treatment leads to a dramatic increase in the cytoskeletal Young's modulus (150-230 kPa) due to protein cross-linking by imine bonds. These results demonstrate the high sensitivity of the single-cell compression method to the molecular-level structural changes of cells, which suggests a new generic platform for investigating cell mechanics in tissue engineering and cancer research.

Cell Shape↗
PubMed1994

Diffusion rates and transport pathways of fluorescein isothiocyanate (FITC)-labeled model compounds through buccal epithelium.

The aim of this study was to characterize transport of FITC-labeled dextrans of different molecular weights as model compounds for peptides and proteins through buccal mucosa. The penetration of these dextrans through porcine buccal mucosa (a nonkeratinized epithelium, comparable to human buccal mucosa) was investigated by measuring transbuccal fluxes and by analyzing the distribution of the fluorescent probe in the epithelium, using confocal laser scanning microscopy for visualizing permeation pathways. The results revealed that passage of porcine buccal epithelium by hydrophilic compounds such as the FITC-dextrans is restricted to permeants with a molecular weight lower than 20 kDa. The permeabilities of buccal mucosa for the 4- and 10-kDa FITC-dextran (of the order of 10(-8) cm/sec) were not significantly different from each other or from the much smaller compound FITC. The confocal images of the distribution pattern of FITC-dextrans showed that the paracellular route is the major pathway through buccal epithelium.

Animals↗
PubMed2004

Nickel content of as-received and retrieved NiTi and stainless steel archwires: assessing the nickel release hypothesis.

This study assesses the nickel content of as-received and retrieved stainless steel and NiTi archwires alloys. New and used brand-matched, composition-matched, and cross section-matched archwires were subjected to scanning electron microscopy and energy-dispersive electron probe microanalysis. Elemental analysis was performed on three randomly selected areas, and the nickel content, expressed as ratios of Ni/Ti (in NiTi wires) or Ni/Fe (in stainless steel), was statistically analyzed with a t-test (alpha = .05). No changes were detected with respect to Ni content ratios between as-received and retrieved NiTi or stainless steel wires, suggesting an absence of nickel release. Wear and delamination phenomena on the wire surface and the formation of galvanic couple between the stainless steel wires and bracket brazing materials intraorally may modify the corrosion susceptibility of the wire alloys in clinical conditions.

Corrosion↗
PubMed2001

Femtosecond near-field spectroscopy: carrier relaxation and transport in single quantum wires.

Quasi-two-colour femtosecond pump and probe spectroscopy and near-field scanning optical microscopy are combined to study the carrier dynamics in single semiconductor nanostructures. In temporally, spectrally and spatially resolved measurements with a time resolution of 200 fs and a spatial resolution of 200 nm, the non-linear change in reflectivity of a single quantum wire is mapped in real space and time. The experiments show that carrier relaxation into a single quantum wire occurs on a 100 fs time scale at room temperature. Evidence is given for a transient unipolar electron transport along the wire axis on a picosecond time and 100 nm length scale.

Journal Article↗
PubMed1995

Increased perinuclear Ca2+ activity evoked by metabotropic glutamate receptor activation in rat hippocampal neurones.

1. The effect of metabotropic glutamate receptor activation on intracellular Ca2+ activity (alpha Cai) of rat hippocampal pyramidal neurones in vitro was examined using ratiometric confocal laser scanning microscopy with the Ca(2+)-sensitive fluorescent probe indo-1 AM. 2. Metabotropic receptors were selectively activated with 1S,3R-1-aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD; 100 microM) in the presence of D-2-amino 5-phosphonovaleric acid (D-APV), 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) and CdCl2. Most pyramidal neurones (77/84) responded with an elevation in Ca2+ activity, maximal after 3-5 min. Fluorescence ratio responses were concentration dependent (EC50 approximately 10 microM) and were blocked by prior application of the antagonist (RS)-4-carboxy-3-hydroxyphenylglycine (RS-CHPG, 300 microM). 3. Responses to 1S,3R-ACPD (100 microM) also caused acidification of the neurones, from estimated control pH 7.2 to pH 6.6 (measured with the pH-sensitive dye SNAFL-calcein). The correction factor for indo-1 determination of Ca2+ was estimated to be x 1.4. 4. Elevations in alpha Cai were greater within the perinuclear region (> 1000 nM), than in the cytoplasm (approximately 200 nM). This region was devoid of staining by the endoplasmic reticulum staining dye 3,3'-dihexyloxacarbocyanine iodide (DiOC6(3)). 5. It is concluded that activation of metabotropic receptors in immature rat hippocampal pyramidal neurones leads to a large increase in perinuclear Ca2+ which would be well positioned to interact with the genome.

Animals↗
PubMed2002

Direct observation of internal spin structure of magnetic vortex cores.

Thin film nanoscale elements with a curling magnetic structure (vortex) are a promising candidate for future nonvolatile data storage devices. Their properties are strongly influenced by the spin structure in the vortex core. We have used spin-polarized scanning tunneling microscopy on nanoscale iron islands to probe for the first time the internal spin structure of magnetic vortex cores. Using tips coated with a layer of antiferromagnetic chromium, we obtained images of the curling in-plane magnetization around and of the out-of-plane magnetization inside the core region. The experimental data are compared with micromagnetic simulations. The results confirm theoretical predictions that the size and the shape of the vortex core as well as its magnetic field dependence are governed by only two material parameters, the exchange stiffness and the saturation magnetization that determines the stray field energy.

Journal Article↗
PubMed1994

Evaluation of Fleroxacin Activity against Established Pseudomonas fluorescens Biofilms.

Scanning confocal laser microscopy (SCLM) and fluorescent molecular probes were used to evaluate the effect of the fluoroquinolone fleroxacin on the architecture of established Pseudomonas fluorescens biofilms. Control P. fluorescens biofilms were heterogeneous, consisting of cell aggregates extending from the attachment surface to maximum measured depths of approximately 90 mum (mean biofilm depth at 72 h, 42 +/- 28 mum) and penetrated by an array of channels. In contrast, fleroxacin-treated biofilms were less deep (mean biofilm depth at 72 h, 29 +/- 8 mum), varied little in depth over large areas, and consisted of a homogeneous distribution of cells. Fleroxacin also caused cells to elongate, with cells located near the biofilm-liquid interface lengthening significantly more than cells located at the attachment surface. By using SCLM, acridine orange, and image analysis it was found that approximately 59% of cells within fleroxacin-treated biofilms emitted red fluorescence whereas >99% of cells from control biofilms emitted green fluorescence. The fleroxacin-treated cells which emitted red fluorescence were observed to be the population of cells which elongated.

Journal Article↗
PubMed1996

Bacterial plasmolysis as a physical indicator of viability.

Bacterial plasmolytic response to osmotic stress was evaluated as a physical indicator of membrane integrity and hence cellular viability. Digital image analysis and either low-magnification dark-field, high-magnification phase-contrast, or confocal laser microscopy, in conjunction with pulse application of a 1.5 M NaCl solution, were used as a rapid, growth-independent method for quantifying the viability of attached biofilm bacteria. Bacteria were considered viable if they were capable of plasmolysis, as quantified by changes in cell area or light scattering. When viable Salmonella enteritidis biofilm cells were exposed to 1.5 M NaCl, an approximately 50% reduction in cell protoplast area (as determined by high-magnification phase-contrast microscopy) was observed. In contrast, heat- and formalin-killed S. enteritidis cells were unresponsive to NaCl treatment. Furthermore, the mean dark-field cell area of a viable, sessile population of Pseudomonas fluorescens cells (approximately 1,100 cells) increased by 50% as a result of salt stress, from 1,035 +/- 162 to 1,588 +/- 284 microns2, because of increased light scattering of the condensed, plasmolyzed cell protoplast. Light scattering of ethanol-killed control biofilm cells underwent little change following salt stress. When the results obtained with scanning confocal laser microscopy and a fluorescent viability probe were compared with the accuracy of plasmolysis as a viability indicator, it was found that the two methods were in close agreement. Used alone or in conjunction with fluorochemical probes, physical indicators of membrane integrity provided a rapid, direct, growth-independent method for determining the viability of biofilm bacteria known to undergo plasmolysis, and this method may have value during efficacy testing of biocides and other antimicrobial agents when nondestructive time course analyses are required.

Bacteria↗
PubMed1996

Structure and function of a nitrifying biofilm as determined by in situ hybridization and the use of microelectrodes.

Microprofiles of O2 and NO3- were measured in nitrifying biofilms from the trickling filter of an aquaculture water recirculation system. By use of a newly developed biosensor for NO3-, it was possible to avoid conventional interference from other ions. Nitrification was restricted to a narrow zone of 50 microns on the very top of the film. In the same biofilms, the vertical distributions of members of the lithoautotrophic ammonia-oxidizing genus Nitrosomonas and of the nitrite-oxidizing genus Nitrobacter were investigated by applying fluorescence in situ hybridization of whole fixed cells with 16S rRNA-targeted oligonucleotide probes in combination with confocal laser-scanning microscopy. Ammonia oxidizers formed a dense layer of cell clusters in the upper part of the biofilm, whereas the nitrite oxidizers showed less-dense aggregates in close vicinity to the Nitrosomonas clusters. Both species were not restricted to the oxic zone of the biofilm but were also detected in substantially lower numbers in the anoxic layers and even occasionally at the bottom of the biofilm.

Biofilms↗
PubMed2005

Quantitative chemical mapping of sodium acrylate- and N-vinylpyrrolidone-enhanced alginate microcapsules.

Alginate microcapsules enclosing recombinant cells secreting therapeutic products have been used successfully to treat several murine models of human diseases. The mechanical and chemical properties of these alginate capsules can be improved by the addition and in situ photo-polymerization of sodium acrylate and N-vinylpyrrolidone in the alginate capsule. The purpose of this modification was to form additional covalent cross-links. In this work we have used scanning transmission X-ray microscopy (STXM) to probe the nature and location of the chemical modifications in the modified capsules by comparison with unmodified capsules. Analysis of X-ray image sequences and selected area spectra has been used to map the calcium gradient in capsules, to identify the presence of polyacrylate throughout the capsules and the localization of poly-N-vinylpyrrolidone in the outer regions of the alginate capsules. The differences in the spatial distributions of these species have led to better understanding of the chemical modifications that provide a mechanically more stable capsule structure.

Acrylates↗
PubMed2005

Effects of gingko biloba extract on glutamate-induced [Ca2+]i changes in cultured cortical astrocytes after hypoxia/reoxygenation, H2O2 or L-glutamate injury.

AIM: To investigate glutamate-induced [Ca2+]i changes in cultured rat neonatal cortical astrocytes after hypoxia/reoxygenation, H2O2 or high concentration of L-glutamate injury. In the meantime, the effects of Gingko biloba extract (GbE) were examined. METHODS: [Ca2+]i changes in astrocytes were monitored by laser scanning confocal microscopy with the Ca2+ sensitive fluorescent probe fluo-3. RESULTS: After astrocytes were impaired by hypoxia/reoxygenation, H2O2 (50 micromol x L(-1)) or L-glutamate (0.25 mmol x L(-)), the exogenous glutamate (27 micromol x L(-1)) could not induce increase of [Ca2+]i, but decrease by (3.3 +/- 1.6)%, (81 +/- 11)% and (81 +/- 7)%, respectively. Pretreatment with GbE (10 mg x L(-1)) could not improve injured astrocytic glutamate response. But after pretreatment with GbE (100 mg x L(-1)), glutamate-induced [Ca2+]i elevation of astrocytes after hypoxia/reoxygenation, H2O2 or high concentration of L-glutamate injury were (135 +/- 98)%, (117 +/- 93)% and (89 +/- 36)%, respectively. Nimodipine (1.6 mg x L(-1)) could also reverse the abnormal response of astrocytes after different injury. CONCLUSION: Hypoxia/reoxygenation, H2O2 and high concentration of L-glutamate impaired astrocytes' response to exogenous L-glutamate, and then bidirectional communication between astrocytes and neurons could not take place. GbE could improve the abnormal responses and maintain the normal function of astroglical network. These effects support that GbE has potential beneficial actions against brain injury.

Animals↗
PubMed2005

[Effects of histamine on proliferation, apoptosis and differentiation of human keratinocytes].

OBJECTIVE: To investigate the effects of histamine on the proliferation, apoptosis and differentiation of human keratinocytes (HKC) and the mechanisms. METHODS: The effect of histamine on the growth of HKC in vitro was examined by MTT assay and trypan blue exclusion assay. Cell cycle analysis and early apoptosis analysis by double staining with Annexin V-FITC and PI were carried out by flow cytometry. DNA ladder assay was performed for the detection of cell apoptosis. HKC free calcium concentration ([Ca(2+)](i)) was measured by laser scanning confocal microscopy in combination with calcium fluorescence probe Fluo-3/AM. HKC differentiation markers keratin 10 (K10) and involucrin was detected by streptavidinbiotin complex immunocytochemical assay. RESULTS: Histamine at high concentration inhibited the proliferation of HKC with cell viability ratio of 65.6% at 1 x 10(-4) mol/L, while histamine at low concentrations promoted proliferation of HKC with the cell viability ratio of 130.7% at 1 x 10(-8) mol/L. Histamine at 1 x 10(-4) mol/L altered cell cycle distribution of HKC with an increase in G0/G1-phase cell population to 30.97%, a decrease in S-phase population to 73.81% and inhibition of G1/S switching. Histamine at 1 x 10(-4) mol/L induced obvious apoptosis of HKC with early apoptosis ratio of 18.64% as compared with the control (5.60%, P<0.05). Histamine 1 x 10(-4) mol/L induced an increase of HKC [Ca(2+)](i) up to 58.9% and cimetidine (an H(2) receptor antagonist) decreased HKC [Ca(2+)](i) down to 25.4%. Histamine at this concentration down-regulated the expressions of K10 and involucrin of HKC but these changes were not significantly different from those of the control (P>0.05). CONCLUSIONS: Histamine at high concentrations inhibits cell cycle progress of HKC, mediates cell apoptosis and induces the increase of [Ca(2+)](i), which might be a partial explanation for growth arrest of HKC elicited by histamine. Histamine may regulate epidermal tissue turnover under physiological conditions, whereas under pathological circumstances of the skin as in trauma and inflammation, histamine at high concentrations may inhibit the regeneration of epidermis and the differentiation of HKC.

Apoptosis↗
PubMed1991

Inorganic components and the fine structures of marginal and deep subgingival calculus attached to human teeth.

Inorganic components and the fine structures of marginal ledge-type and deep subgingival spiny deposits in human old dental calculus were investigated by scanning electron microscopy and energy dispersive electron-probe microanalysis. The ledge-type deposits consisted of the extra- and intracellular calcifying deposits, large plate-shaped crystals, and bacillus-shaped deposits composed of hexahedrally based crystals. The spiny deposits were mainly formed by aggregations of the bacillus-shaped deposits. In the outer and middle layers of the spiny deposits, the Ca, P, and Mg concentrations were all significantly higher than those of the ledge-type deposits. A consideration of the crystal shapes and Ca, P, and Mg molar ratios reveals the following differences. Calculus components of the ledge-type deposits contained crystal types quite similar to sandy grain-shaped hydroxyapatite (HAP), plate-shaped octacalcium phosphate (OCP), and hexahedral Mg-containing whitlockite (WHT). On the other hand, in the spiny deposits, the Mg-containing WHT type comprised a large proportion of the calculus; the HAP type was found in the outermost and inner layers; and no OCP type was detected.

Calcium Phosphates↗
PubMed1994

X-ray scanning microanalysis of Sporothrix schenckii.

X-ray scanning microanalysis was applied to the study of fungal samples such as Sporothrix schenckii. This technique is sufficiently sensitive to detect many elements in the periodical classification of Mendeleev.

Electron Probe Microanalysis↗
PubMedDate not supplied

[The characteristics of osteoclast localization in the bones of the extremities of the crested salamander Pleurodeles waltlii].

Localization of polynuclear osteoclasts in limb bones of ribbed newt were analyzed in consideration of calcium content in different bone structures. With using the methods of light and scanning electron microscopy and X-ray microanalysis it was shown that a critical level of cartilage calcification is need to activate osteoclasts. Osseous tissue can bind more calcium salt without resorption.

Animals↗
PubMed1996

Sequence-specific labeling of superhelical DNA by triple helix formation and psoralen crosslinking.

Site-specific labeling of covalently closed circular DNA was achieved by using triple helix-forming oligonucleotides 10, 11 and 27 nt in length. The sequences consisted exclusively of pyrimidines (C and T) with a reactive psoralen at the 5'-end and a biotin at the 3'-end. The probes were directed to different target sites on the plasmids pUC18 (2686 bp), pUC18/4A (2799 bp) and pUC1 8/4A-H 1 (2530 bp). After triple helix formation at acid pH the oligonucleotides were photocrosslinked to the target DNAs via the psoralen moiety, endowing the covalent adduct with unconditional stability, e.g. under conditions unfavorable for preservation of the triplex, such as neutral pH. Complex formation was monitored after polyacrylamide gel electrophoresis by streptavidin-alkaline phosphatase (SAP)-induced chemiluminescence. The yield of triple helix increased with the molar ratio of oligonucleotide to target and the length of the probe sequence (27mer > 11mer). The covalent adduct DNA were visualized by scanning force microscopy (SFM) using avidin or streptavidin as protein tags for the biotin group on the oligonucleotide probes. We discuss the versatility of triple helix DNA complexes for studying the conformation of superhelical DNA.

Base Sequence↗