1,878 research outputs found
Evaluation of the hCMEC/D3 cell line, a new "in vitro" model of the human blood-brain barrier for transport and gene regulation studies
No full textBrain endothelial capillary cells form the blood-brain barrier (BBB), a highly selective
membrane between the peripheral blood and the central nervous system. The main
functions of the BBB are to protect the brain tissue by preventing the entry of toxic
compounds and to supply it with nutrients in order to assure proper function. Tight
junctions are the key elements for the establishment of a tight barrier and seal the
intercellular gaps against passive diffusion of hydrophilic compounds. A second important
characteristic of the brain capillary endothelial cells are transport proteins that prevent
brain penetration of their substrates by pumping them back in the blood. These
compounds include a series of clinically used drugs. Important drug efflux transporters
located at the BBB are P-glycoprotein (P-gp), the breast cancer resistance protein
(BCRP) and the family of multidrug resistance proteins (MRP).
During drug development, the question of whether a drug candidate reaches the brain
tissue is of great importance. Therefore, models are needed to predict the BBB
permeability of new compounds. In the past, in vitro models have been developed to
address this question. These models include isolated brain capillaries, isolated primary
brain capillary endothelial cells and BBB cell lines of various origins. A major problem
encountered with these cell lines was an insufficient paracellular resistance.
Recently, the hCMEC/D3 cell line was generated by immortalizing primary human brain
endothelial cells. In culture this cell line shows a morphology that closely resembles to
primary cells, forms tight monolayers and expresses BBB markers such as chemokine
receptors, tight junctional molecules and ATP binding cassette (ABC)-transporters.
The aim of this thesis was to evaluate the hCMEC/D3 cell line as an in vitro model of the
human BBB to study 1) permeability properties including para- and transcellular diffusion
as well as active transport 2) the influence of endo- and exogenous factors on the
paracellular permeability and 3) the regulation of breast cancer resistance protein and Pglycoprotein
by pro-inflammatory cytokines.
The first study describes the characterization of the hCMEC/D3 cells as an in vitro model
of the human BBB for permeability studies (section Error! Reference source not
found.). The ability of the cells to allow discrimination between para- and transcellular
diffusion was investigated by measuring the transport of a series of compounds with
different physicochemical properties. A ratio of 2.8 was observed when comparing the
permeabilities of the compounds with the highest and the lowest diffusion rate. The
passive permeability of sucrose could be reduced significantly by replacing fetal calf
serum with human serum. Furthermore, quantitative mRNA expression of the ABCtransporters
P-gp, BCRP, MRP1, MRP2, MRP3, MRP4, MRP5 as well as the human
transferrin receptor (hTfR) was shown. Protein expression of P-gp, BCRP and the hTfR
was detected and functional activity of P-gp, BCRP and the MRPs was investigated in
efflux experiments. Furthermore, bidirectional P-gp transport activity was observed.
In a second project the impact of endo- and exogenous factors on the paracellular
permeability of hCMEC/D3 monolayers was assessed, since it is know that the molecular
assembly of tight junctions depends on the surrounding milieu (section Error! Reference
source not found.). Based on reports in the literature, the cells were incubated with a
variety of compounds that included anti-inflammatory drugs, growth factors and
antioxidants. The effects on the monolayer tightness of hCMEC/D3 were investigated by
measuring the transport of sucrose, a paracellular permeability marker. N-acetylcystein
(NAC), atorvastatin and sodium nitroprusside (SNP) reduced the sucrose permeability
significantly, and slightly increased zonula occludens protein (ZO-1) expression.
Additionally, NAC and SNP reduced the generation of reactive oxygen species (ROS),
which have been reported to disrupt the assembly of tight junctions.
The effect of the pro-inflammatory cytokines IL-1[beta], IL-6 and TNF-[alpha] on the expression and
activity of the ABC-transporters BCRP and P-gp was investigated in the hCMEC/D3 cell
line (section Error! Reference source not found.). IL-1[beta], IL-6 and TNF-[alpha], which are
know to be elevated during various diseases, suppressed significantly BCRP mRNA
expression. In addition, BCRP activity was reduced under the influence of all tested
cytokines, as shown by efflux experiments. P-gp mRNA levels were slightly reduced by
IL-6 but significantly increased after TNF-[alpha] treatment. TNF-[alpha] also increased the protein
expression of P-gp. This in vitro study indicates that expression levels of BCRP and P-gp
at the BBB might be altered during acute or chronic inflammation, resulting in a changed
brain penetration of their substrates.
In an isolated project, the pharmacokinetics and pharmacodynamics of increasing oral
doses of the satiety peptides GLP-1 and PYY3-36 were assessed in healthy male
volunteers. Oral administration of either peptide induced a rapid and dose-dependent
increase in plasma drug concentrations. Oral administration of GLP-1 induced a potent
effect on insulin release and both peptides suppressed ghrelin secretion. In conclusion,
this study showed, for the first time, that satiety peptides such as GLP-1 and PYY3-36
can be orally delivered safely and effectively in humans
Birk, H E, 402634
No full textThis record was harvested from a previous catalogue system and will be withdrawn in 2025. Information in this record may be superseded or incomplete. Visit this record in UMA's new catalogue at: https://archives.library.unimelb.edu.au/nodes/view/371905Surname: BIRK
Given Name(s) or Initials: H E
Military Service Number or Last Known Location: 402634
Missing, Wounded and Prisoner of War Enquiry Card Index Number: 7556182825
Item: [2016.0049.04232] "Birk, H E, 402634
The amino acid sequence of a cereal Bowman-Birk type trypsin inhibitor from seeds of Jobs' tears (Coix lachryma-jobi L.)
No full textAbstractThe major trypsin inhibitor from seeds of Jobs' tears (Coix lachryma-jobi) was purified by heat treatment, fractional precipitation with (NH4)2SO4, ion-exchange chromatography on DEAE-Sepharose, gel-filtration on Sephadex G-75 and preparative reverse-phase HPLC. The complete amino acid sequence was determined by analysis of peptides derived from the reduced and S-carboxymethylated protein by digestion with trypsin, chymotrypsin and the S. aureus V8 protease. The polypeptide contained 64 amino acids with a high content of cysteine. The sequence exhibited strong homology with a number of Bowman-Birk inhibitors from legume seeds and similar proteins recently isolated from wheat and rice
Retracing prairie voices: a conversation with Birk Sproxton
No full textRetracing prairie voices : a conversation with Birk Sproxton. - In: Prairie fire. 8. 1987. 2. S. 4-1
Arbeitslosigkeit und politisches Engagement in den Neuen Bundesländern
No full textArbeitslosigkeit und politisches Engagement in den Neuen Bundesländern / N. Birk ; J. König ; Ph. Mayring. - In: Zeitschrift für politische Psychologie. 1994. S. 213-237. - Auch ersch. als: Augsburger Berichte zur Entwicklungspsychologie und pädagogischen Psychologie 6
Arbeits- und sozialversicherungsrechtliche Folgen einer fehlerhaften Prognose des Entbindungszeitpunktes
No full textArbeits- und sozialversicherungsrechtliche Folgen einer fehlerhaften Prognose des Entbindungszeitpunktes / Rolf Birk ; Siegfried Deffner. - In: Die Sozialgerichtsbarkeit. 27. 1980. S. 6-1
Zur Problematik multinationaler Unternehmen
No full textZur Problematik multinationaler Unternehmen / Rolf Birk ; Hans Tietmeyer. - Paderborn: Schöningh, 1976. - 60 S. - (Rechts- und staatswissenschaftliche Veröffentlichungen der Görres-Gesellschaft ; N.F., 25
Wie funktioniert das? Die Wirtschaft heute
No full textWie funktioniert das? : Die Wirtschaft heute / Rolf Birk ... - Mannheim: Bibliogr. Inst., 1976. - 704 S
Identification of localised recharge and conduit flow by combined analysis of hydraulic and physico-chemical spring responses (Urenbrunnen, SW-Germany)
No full textKarst aquifers are highly vulnerable to contamination due to the rapid transport of pollutants in conduit systems. Effective strategies for the management and protection of karst aquifers, therefore, require an adequate hydrogeological characterisation of the conduit systems. In particular, the identification and characterisation of conduits transmitting rapid, localised recharge to springs is of great interest for vulnerability assessments. In this work, it is demonstrated that localised recharge and conduit flow in a karst aquifer (Urenbrunnen catchment, southwest Germany) can be characterised by jointly analysing the hydraulic and physico-chemical responses of a spring to recharge events. Conduit volumes are estimated by evaluating time lags between increases in spring discharge and associated changes in the electrical conductivity and temperature of the discharged water. These estimates are confirmed by the results of a combined tracer and recharge test. Variations in electrical conductivity are also shown to assist in the quantification of the fast recharge component associated with short-term recharge pulses. However, spectral analysis of temperature fluctuations reveals that highly mineralised surface waters locally infiltrate into the aquifer during the winter and spring without causing significant electrical conductivity variations in the spring water. Hence, the most consistent conceptual model is obtained by a combined analysis of both physico-chemical parameters. (C) 2003 Elsevier B.V. All rights reserved
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