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  • 1
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature America, Inc.
    Nature genetics 23 (1999), S. 34-35 
    ISSN: 1546-1718
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] The development of novel pharmaceutically active compounds requires effective methods in order to increase throughput and focus corporate research and development. The increasing number of potential compounds in pharmaceutical research and the shortened time frame in the drug development have ...
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1420-9071
    Keywords: Key words. Phagocytosis; cyclosporine A particles; morphometry; latex particles.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. The aim of this study was to develop an in vitro model to estimate the clearance of pulmonary administered cyclosporine A (CsA). To do this we estimated the volume of CsA particles phagocytosed by alveolar macrophages (AM) lavaged from hamsters. AM were cultured with CsA particles at two doses of particles (0.1 mg or 0.5 mg) and at three incubation times (1 h, 6 h or 24 h). The AM were also incubated with or without latex particles. After incubation, AM were processed for light and electron microscopy and the mean volume of phagocytosed particles was esti mated stereologically from micrographs of the cells. Here, however, the CsA particles were dissolved during the embedding process and only their negative images (vacuoles) could be detected. An indirect method was therefore developed. The volume of cytoplasmic vacuoles (called 'background' vacuoles) was estimated in control macrophages (without particles or with latex particles and subtracted from the total volume of vac uoles in macrophages incubated with CsA, which gave the volume of phagocytosed CsA. The volume of the 'background' vacuoles remained constant in all study conditions. At a dose of 0.1 mg CsA the volume phagocytosed per macrophage was 13.83 μm3 at 1 h, 8.43 μm3 at 6 h and 4.50 μm3 at 24 h. At a dose of 0.5 mg CsA, the volume phagocytosed varied from 26.59 μm3 at 1 h, to 4.13 μm3 at 6 h and 49.10 μm3 at 24 h. These results show no statistically significant dependence on time for either dose, and a statistically significant dose effect only at 24 h. With latex particles, the phagocytosed volume increased significantly with time and dose and was significantly higher than for CsA particles. This study showed that CsA particles are phagocytosed by AM from hamsters but to a lesser extent than latex particles. This difference could be correlated with physical properties, i.e. a difference between particle size and shape and/or chemical properties, latex particles being inert and CsA particles being peptidic. Moreover, different surface receptors on AM could be involved in the process of phagocytosis of CsA and latex particles.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-904X
    Keywords: octreotide ; somatostatin analogue ; SMS 201-995 ; peptides ; blood–brain barrier ; transport ; visualization ; confocal laser scanning microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Abstract Confocal laser scanning microscopy (CLSM) was used to quantify and visualize the transport of the octapeptide and somatostatin analogue, octreotide (SMS 201-995, Sandostatin), across monolayers of bovine cerebrovascular endothelial cells, an in vitro model of the blood–brain barrier. The concentrations of octreotide and its conjugates in the cell culture medium were determined by radioimmunoassay (RIA). Two fluorescent conjugates of octreotide (FITC- and NBD-octreotide) were used to obtain CLSM images. The peptides did not undergo significant degradation in the presence of brain endothelial cell monolayers. The transport rate of octreotide expressed as clearance (Cl) and endothelial permeability (P e) did not depend on either the initial concentration (between 10 nM and 1 µM) or the site of administration (luminal or abluminal side of the mono-layer), indicating the absence of saturable and/or asymmetrical transport mechanisms. The P e of octreotide and that of the paracellular permeability marker fluorescein correlated well. Although the conjugates are more lipophilic than octreotide itself, they exhibited lower Cl and P e, values probably because of their larger molecular size. On the CLSM images, FITC-octreotide was present only in the intercellular space, while the cells did not exhibit detectable fluorescence. Transport studies and CLSM images suggest that octreotide passes the endothelial monolayer primarily via the paracellular route without significant contribution of carrier-mediated transport.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1573-6822
    Keywords: cell activation ; cell viability ; cyclosporin ; phagocytosis ; stereology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The aim of this study was to compare two types of cyclosporin (Cs) particles, SDZ OXL 400 and SDZ IMM 125, the latter being more hydrophilic, to understand their uptake by airway macrophages. Alveolar macrophages (AM), harvested by bronchoalveolar lavage (BAL) of hamster lungs, were cultured with two different doses (0.1 mg and 0.5 mg) for 1 h, 6 h, and 24 h. Control incubations without Cs particles or with latex particles were carried out simultaneously. Cell viability, cell activation (i.e., respiratory burst, interleukin-6 (IL-6) synthesis) and mean volume of particles phagocytosed per macrophage were measured. Both types of Cs particles did not modify the AM viability, and failed to induce IL-6 synthesis during phagocytosis but slightly decreased the cell oxidative respiratory burst. The comparison between SDZ OXL 400 and SDZ IMM 125 showed that for the lower dose the mean volume of both Cs types phagocytosed was similar at 1 h and 6 h. At 24 h an increase of the mean volume phagocytosed was seen for SDZ IMM 125 but not for SDZ OXL 400. For the higher dose the mean volume of SDZ IMM 125 phagocytosed was higher than SDZ OXL 400 at 1 h and 6 h and comparable for both types at 24 h. SDZ IMM 125 particles were phagocytosed more rapidly than SDZ OXL 400. The mean volume of phagocytosed latex particles increased with time and dose and was higher than for both Cs particle types. In conclusion, AM were seen to phagocytose particles of different physical properties (i.e., form, size, and shape), chemical properties (i.e., inert or peptidic) and degrees of hydrophilicity in a different manner.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1573-6822
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1573-6822
    Keywords: human ; in vitro ; nasal mucosa ; octreotide ; Sandostatin®
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Anin vitro human nasal model was developed as a tool to study the local tolerabiliity of nasal powder forms using excised nasal mucosa in a diffusion chamber. The suitability of this model was tested using Sandostatin® (SMS) an octapeptide analog of somatostatin, as a reference drug enhanced by Avicel® (microcrystalline cellulose) or lactose (100 mesh). The standard nasal spray vehicle was taken as a harmless control and 1% chenodeoxycholate (CDC) as a harmful control in terms of local tolerability. The extent of peptide permeation was determined by measuring SMS concentration in the receiving chamber. The labeling of SMS was detected by immunoperoxidase staining on cross sections. The local tolerability for all tested forms was assessed by histopathological examination and scanning electron microscopy. The apparent permeation coefficient allowed us to rank the absorption of the tested drug forms as Avicel 〉 spray=lactose〉1%CDC. For all formulations, SMS was detected in the epithelium. No changes of the nasal mucosa could be observed with Avicel, lactose or nasal spray vehicle in the presence or absence of SMS. 1%CDC with or without drug showed an immediate destruction of the nasal epithelium. The validation of thisin vitro model using human nasal mucosa will be further discussed as a tool for assessing the local tolerability of intranasally applied test substances.
    Type of Medium: Electronic Resource
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