Impact of ATP-binding cassette, subfamily B, member 1 pharmacogenetics on tacrolimus-associated nephrotoxicity and dosage requirements in paediatric patients with liver transplant

Importance of the field: Tacrolimus is the most commonly used immunosuppressive agent following solid-organ transplantation in children. Its clinical use, however, is complicated by side effects (mainly nephrotoxicity), narrow therapeutic index and pharmacokinetic variability which can result in an increased risk of treatment failure or toxicity. Studies examining interindividual differences in the expression of the ABCB1 (ATP-binding cassette, subfamily B, member 1) gene (which encodes the drug transporter, P-gp) and its genetic polymorphisms have attempted to elucidate variations in tacrolimus response and disposition in children. Areas covered in this review: This review explores pharmacogenetic knowledge developed over the last decade regarding the impact of ABCB1 polymorphisms on tacrolimus toxicity and dosage requirements in children. What the reader will gain: A better understanding of the role of ABCB1 genetic polymorphisms (and corresponding haplotypes) and ABCB1 expression levels in various tissues and organs on tacrolimus outcomes in children with liver transplant. Take home message: Pharmacogenetics offers significant potential for optimising tacrolimus use. ABCB1 donor genotypes and ABCB1 expression level in the intestine and leukocytes may be useful in dosage selection. Large prospective studies are, however, required to further explore the potential of genetic testing in identifying children who are at risk of toxicity and to better individualise tacrolimus therapy.

Detergent-free purification of ABC (ATP-binding-cassette) transporters

ABC (ATP-binding-cassette) transporters carry out many vital functions and are involved in numerous diseases, but study of the structure and function of these proteins is often hampered by their large size and membrane location. Membrane protein purification usually utilizes detergents to solubilize the protein from the membrane, effectively removing it from its native lipid environment. Subsequently, lipids have to be added back and detergent removed to reconstitute the protein into a lipid bilayer. In the present study, we present the application of a new methodology for the extraction and purification of ABC transporters without the use of detergent, instead, using a copolymer, SMA (polystyrene-co-maleic acid). SMA inserts into a bilayer and assembles into discrete particles, essentially solubilizing the membrane into small discs of bilayer encircled by a polymer, termed SMALPs (SMA lipid particles). We show that this polymer can extract several eukaryotic ABC transporters, P-glycoprotein (ABCB1), MRP1 (multidrug-resistance protein 1; ABCC1), MRP4 (ABCC4), ABCG2 and CFTR (cystic fibrosis transmembrane conductance regulator; ABCC7), from a range of different expression systems. The SMALP-encapsulated ABC transporters can be purified by affinity chromatography, and are able to bind ligands comparably with those in native membranes or detergent micelles. A greater degree of purity and enhanced stability is seen compared with detergent solubilization. The present study demonstrates that eukaryotic ABC transporters can be extracted and purified without ever being removed from their lipid bilayer environment, opening up awide range of possibilities for the future study of their structure and function. © The Authors Journal compilation © 2014 Biochemical Society.

Systems biology approach to study permeability of paracetamol and its solid dispersion

Physiological changes that take place at cellular level are usually reflective of their level of gene expression. Different formulation excipients have an impact on physiological behavior of the exposed cells and in turn affect transporter genes, enterocyte-mediated metabolism and toxicity biomarkers. The aim of this study was to prepare solid dispersion of paracetamol and evaluate genetic changes that occur in Caco-2 cell lines during the permeability of paracetamol alone and paracetamol solid dispersion formulations. Paracetamol-PEG 8000 solid dispersion was prepared by melt fusion method and the formulation was characterised using differential scanning calorimetry (DSC), scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). Formulation of solid dispersion resulted in the conversion of crystalline drug into an amorphous form. Permeability studies showed that paracetamol absorption was higher from the solid dispersion formulation. DNA microarrays analysis was carried out in order to investigate the involvement of any efflux/uptake transporters in paracetamol or its solid dispersion permeability. Neither transporter carriers nor efflux proteins were found to be involved in the absorption of paracetamol or its PEG solid dispersion. Gene expression analysis established that paracetamol toxicity was potentially reduced upon formulation into solid dispersion when ATP binding cassette (ABC) and solute carrier transporter (SLC) genes were analyzed.

Humans assume a mixture of diffuse and point-source lighting when viewing sinusoidal shading patterns

Observers perceive sinusoidal shading patterns as being due to sinusoidally corrugated surfaces, and perceive surface peaks to be offset from luminance maxima by between zero and 1/4 wavelength. This offset varies with grating orientation. Physically, the shading profile of a sinusoidal surface will be approximately sinusoidal, with the same spatial frequency as the surface, only when: (A) it is lit suitably obliquely by a point source, or (B) the light source is diffuse and hemispherical--the 'dark is deep' rule applies. For A, surface peaks will be offset by 1/4 wavelength from the luminance maxima; for B, this offset will be zero. As the sum of two same-frequency sinusoids with different phases is a sinusoid of intermediate phase, our results suggest that observers assume a mixture of two light sources whose relative strength varies with grating orientation. The perceived surface offsets imply that gratings close to horizontal are taken to be lit by a point source; those close to vertical by a diffuse source. [Supported by EPSRC grants to AJS and MAG].

The role of local luminance amplitude in the interpretation of shape-from-shading in textured surfaces

Luminance changes within a scene are ambiguous; they can indicate reflectance changes, shadows, or shading due to surface undulations. How does vision distinguish between these possibilities? When a surface painted with an albedo texture is shaded, the change in local mean luminance (LM) is accompanied by a similar modulation of the local luminance amplitude (AM) of the texture. This relationship does not necessarily hold for reflectance changes or for shading of a relief texture. Here we concentrate on the role of AM in shape-from-shading. Observers were presented with a noise texture onto which sinusoidal LM and AM signals were superimposed, and were asked to indicate which of two marked locations was closer to them. Shape-from-shading was enhanced when LM and AM co-varied (in-phase), and was disrupted when they were out-of-phase. The perceptual differences between cue types (in-phase vs out-of-phase) were enhanced when the two cues were present at different orientations within a single image. Similar results were found with a haptic matching task. We conclude that vision can use AM to disambiguate luminance changes. LM and AM have a positive relationship for rendered, undulating, albedo textures, and we assess the degree to which this relationship holds in natural images. [Supported by EPSRC grants to AJS and MAG].

The fractionation of dextran polymer by ultrafiltration to yield clinical products

A review of ultrafiltration (UF) theory and equipment has been made. Dextran is fractionated industrially by ethanol precipitation, which is a high energy intensive process. The aims of this work were to investigate the fractionation of dextran using UF and to compare the efficiency and costs of UF fractionation with ethanol fractionation. This work is the continuation of research conducted at Aston, which was concerned with the fractionation of dextran using gel permeation chromatography (GPC) and hollow fibre UF membranes supplied by Amicon Ltd. Initial laboratory work centred on determining the most efficient make and configuration of membrane. UF membranes of the Millipore cassette configuration, and the DDS flat-sheet configuration, were examined for the fracationation of low molecular weight (MW) dextran. When compared to Amicon membranes, these membranes were found to be inferior. DDS membranes of 25 000 and 50 000 MW cut-offs were shown to be capable of fractionating high MW dextran with the same efficiency as GPC. The Amicon membranes had an efficiency comparable to that of ethanol fractionation. To increase this efficiency a theoretical UF membrane cascade was adopted to utilize favourable characteristics encountered in batch mode membrane experiments. The four stage cascade used recycled permeates in a counter- current direction to retentate flow, and was operated 24 hours per day controlled by a computer. Using 5 000 MW cut-off membranes the cascade improved the batch efficiency by at least 10% for a fractionation at 6 000 MW. Economic comparisons of ethanol fractionation, combined GPC and UF fractionation, and UF fractionation of dextran were undertaken. On an economic basis GPC was the best method for high MW dextran fractionation. When compared with a plant producing 100 tonnes pa of clinical dextran, by ethanol fractionation, a combined GPC and UF cascade fractionation could produce savings on operating costs and an increased dextran yield of 5%.

Protein-mediated isolation of plasmid DNA by a zinc finger-glutathione S-transferase affinity linker

The sequence-specific affinity chromatographic isolation of plasmid DNA from crude lysates of E. coli DH5α fermentations is addressed. A zinc finger-GST fusion protein that binds a synthetic oligonucleotide cassette containing the appropriate DNA recognition sequence is described. This cassette was inserted into the Smal site of pUC19 to enable the affinity isolation of the plasmid. It is shown that zinc finger-GST fusion proteins can bind both their DNA recognition sequence and a glutathione-derivatized solid support simultaneously. Furthermore, a simple procedure for the isolation of such plasmids from clarified cell lysates is demonstrated. Cell lysates were clarified by cross-flow Dean vortex microfiltration, and the permeate was incubated with zinc finger-GST fusion protein. The resulting complex was adsorbed directly onto glutathione-Sepharose. Analysis of the glutathione-eluted complex showed that plasmid DNA had been recovered, largely free from contamination by genomic DNA or bacterial cell proteins. © 2002 Wiley Periodicals, Inc.

Sammelsurium zum Thema Mauerfall und Wende:Die heterogenen Beiträge eines von Ute Dettmar und Mareile Oetken herausgegebenen Tagungsbandes erforschen kulturelle Reflexe vom Ende der DDR

Review of: Ute Dettmar / Mareile Oetken (Hg.): Grenzenlos. Mauerfall und Wende in (Kinder- und Jugend-)Literatur und Medien. Universitätsverlag Winter, Heidelberg 2010. Pünktlich zum zwanzigsten Jubiläum der Deutschen Einheit legten Ute Dettmar und Mareile Oetken einen Sammelband vor, dessen Beiträge vielen unterschiedlichen Aspekten des Themas „Mauerfall und Wende“ gewidmet sind und deren Autoren aus unterschiedlichen Fachrichtungen stammen. Beste Voraussetzungen für einen wahrhaft interdisziplinären Band, so könnte man meinen, doch diese Lesererwartung wird leider enttäuscht. Zu unvermittelt stehen die Beiträge dieses Konferenzbandes nebeneinander und befassen sich zu deutlich mit ganz unterschiedlichen Fragestellungen, die sich nur sehr begrenzt gegenseitig befruchten. Die ansonsten sehr anregende Sammlung „Grenzenlos“ leidet unter diesem Manko, das man durch eine sinnvolle Einordnung der Beiträge in den weiteren Kontext hätte mindern können. Der geeignete Ort dafür wäre die Einleitung. Diese allerdings fällt enttäuschend knapp aus und trägt nur wenig dazu bei, die zum Teil willkürlich wirkende Heterogenität der Beiträge zu rechtfertigen und übergreifende Ergebnisse zu formulieren. Obwohl die Gesamtkonzeption von „Grenzenlos“ deshalb kaum überzeugt, bietet das Buch erfreulicherweise eine Vielzahl interessanter Einzelbeiträge, die zu einem Großteil aus der Kinder- und Jugendbuchforschung stammen, der Disziplin, in der auch die beiden Herausgeberinnen tätig sind. Wie hier erneut bewiesen wird, lässt sich dieses Untergebiet nur unscharf von der klassischen Literaturwissenschaft trennen. Es lohnt sich, an Texte, die sich an junge Leser richten, kaum modifizierte philologische Maßstäbe anzulegen, denn viele der hier besprochenen Werke lassen in Bezug auf die Auseinandersetzung mit den politischen Ereignissen ähnliche Tendenzen erkennen, wie sie sich in der zum Vergleich herangezogenen „Erwachsenenliteratur“ und anderen Medien, etwa dem Fernsehfilm, beobachten lassen. Diese Erkenntnis betont Carsten Gansel in seinem überblicksartigen Aufsatz. Er zeigt zunächst Tendenzen in der Auseinandersetzung mit der Mauerfallthematik in der Gegenwartsliteratur auf und illustriert dann, wie stark sich Kinder- und Jugendliteratur zum Thema Deutsche Einheit an denselben Aspekten orientiert und ähnliche Erinnerungsdiskurse abbildet. Abschließend beklagt Gansel zwar die in einigen Texten vorherrschenden Klischees, seine Hauptkritik gilt jedoch der unreflektierten Übernahme dieser stereotypen Darstellungen im Schulunterricht. Wie Gansel zu Recht bemängelt und am Beispiel von Anne C. Voorhoeves Jugendbuch „Lilly unter den Linden“ belegt, werden nicht selten literarische Texte als Grundlage einer Diskussion über die Zustände in der DDR herangezogen, ohne dass die Fiktionalität der Bücher ausreichend betont und ihre Diskursivität klar herausgestellt und thematisiert wird. Mit der Konstruktion von Inhalten des „kommunikativen Gedächtnisses“ (Jan Assmann) setzen sich verständlicherweise noch viele weitere Aufsätze des Bandes auseinander. Eine der Fragen, die dabei aufgeworfen worden, ist natürlich, inwiefern der Osten und der Westen unterschiedliche Erinnerungsdiskurse hervorbringen. In seinem sozialwissenschaftlichen Beitrag, der sich unter den literaturwissenschaftlichen Aufsätzen leider etwas wie ein Fremdkörper ausnimmt, betont Thomas Ahbe, wie stark sich Ostdeutsche von den öffentlichen Repräsentationsformen und der Wendenarrativen im vereinten Deutschland ausgeschlossen fühlen. Dem entspricht auf literarischer Seite der zehn Jahre nach der Wiedervereinigung zu beobachtende Trend der Erinnerungsbücher, mit dem sich verständlicherweise mehrere Beiträge des Bandes auseinandersetzen. Wie exemplarisch am Beispiel von Jana Hensels Bestseller „Zonenkinder“ gezeigt wird, schrieb hier aber nicht nur der Osten gegen den Westen an, der mit Florian Illies’ „Generation Golf“ längst ein stilbildendendes Vorbild für das Genre geschaffen hatte. Vielmehr meldete sich zugleich auch eine ostdeutsche Generation zu Wort, die anders als ihre an Kontinuität gewöhnten gleichaltrigen Pendants im Westen und anders als ihre in den entschwundenen Bezugsgrößen der DDR verhafteten Eltern mit einem Empfinden der doppelten Entwurzelung zu kämpfen hatte und um eine feste Identität rang. Das bei Hensel zentral artikulierte Konzept eines verschwundenen Landes im Osten und der damit verbundenen Illegitimierung von Erinnerungs- und Identitätsbildungsprozessen ist jedoch nicht nicht auf die Generation der Zonenkinder beschränkt. Wie Jens Thiele eindrucksvoll belegt, zeigt sich beispielsweise in den Bilderbüchern des ostdeutschen Illustrators Klaus Ensikat erst in den nach der Wende entstandenen Zeichnungen das Lokalkolorit der DDR in den 1970er- und 1980er-Jahren. Wo zuvor leere Hintergründe vorherrschten, wird nun ein zeitgeschichtlicher Kontext abgebildet, der sich in seiner Vieldeutigkeit zwar nicht auf eine verspätete DDR-Darstellung reduzieren lässt, der aber einen Beitrag leistet zu den kulturellen Reflexen dieses verschwundenen Staates. Den Abschluss von „Grenzenlos“ bilden drei aufschlussreiche Aufsätze, die sich der Außenperspektive auf die Geschehnisse in Deutschland und der potenziell daraus resultierenden anderen Wahrnehmung der Nation widmen. Aus britischer, niederländischer und polnischer Sicht stellen sich Mauerfall und Wende anders dar und geben in manchen Fällen Anlass zu einem revidierten Heteroimage der Deutschen. Gestützt auf imagologische Konzepte untersucht Emer O’Sullivan, ob und wie sich die Repräsentation der Deutschen in der britischen Literatur, insbesondere in Kinder- und Jugendbüchern, seit der Einheit verändert hat. Sie gelangt zu dem ernüchternden Schluss, dass die neuere deutsche Geschichte auf die Briten entschieden weniger Faszination ausübt als die in der Literatur und anderen Medien immer wieder dargestellten Weltkriege. Einer weiterhin anwachsenden Anzahl von Büchern für Kinder und Jugendliche, die sich mit Themen wie den deutschen Luftangriffen auf London oder dem Schicksal deutscher Juden (besonders den Erlebnissen der nach Großbritannien geretteten Passagiere der Kindertransporte) beschäftigen, steht eine verschwindend kleine Menge an literarischen oder kinematografischen Auseinandersetzungen mit den Ereignissen von 1989 und ihren Folgen gegenüber. Wer sich in Großbritannien etwas auskennt, den mag dieser Befund nicht erstaunen. Umso interessanter und ermutigender ist es, aus den Aufsätzen von Kirsten Waterstraat und Jacek Rzeszotnik zu erfahren, dass sich das Deutschlandbild in den Niederlanden und in Polen durchaus im Wandel befindet – wenngleich auch dort einige lang tradierte Stereotype keine Ablösung erfahren haben und beispielsweise die spekulative Literatur der polnischen alternate history-Romane weiterhin vorzugsweise das Nazireich in den Vordergrund stellt. Insgesamt ergibt sich bei der Lektüre dieses Bandes ein facettenreiches Bild des Forschungsstandes. So fragmentiert die Anordnung der Beiträge ist, so reichhaltig präsentieren sich die Ergebnisse dieser heterogenen Studien. Es wird deutlich, dass die Kinder- und Jugendliteratur im In- wie im Ausland sich der Mauerfall-Thematik zwar schon mit reichhaltigen Ergebnissen zugewandt hat. Eine differenzierte, von Stereotypen freie Darstellung des Themas, wie auch ein junges Publikum sie zu Recht erwarten kann, ist aber bisher eher selten. Auch eine stärkere Auseinandersetzung mit der gegenwärtigen Lebenswelt im vereinten Deutschland, die der soziopolitischen Entwicklung vom Mauerfall bis heute mehr Aufmerksamkeit schenkt, wird von mehreren der in hier vertretenen Wissenschaftler angemahnt und stellt sicher nicht nur aus der Perspektive dieses Bandes ein Desiderat dar.

Genetic risk factors and age-related macular degeneration (AMD)

Age related macular degeneration (AMD) is the leading cause of blindness in individuals older than 65 years of age. It is a multifactorial disorder and identification of risk factors enables individuals to make lifestyle choices that may reduce the risk of disease. Collaboration between geneticists, ophthalmologists, and optometrists suggests that genetic risk factors play a more significant role in AMD than previously thought. The most important genes are associated with immune system modulation and the complement system, e.g., complement factor H (CFH), factor B (CFB), factor C3, and serpin peptidase inhibitor (SERPING1). Genes associated with membrane transport, e.g., ATP-binding cassette protein (ABCR) and voltage-dependent calcium channel gamma 3 (CACNG3), the vascular system, e.g., fibroblast growth factor 2 (FGF2), fibulin-5, lysyl oxidase-like gene (LOXL1) and selectin-P (SELP), and with lipid metabolism, e.g., apolipoprotein E (APOE) and hepatic lipase (LIPC) have also been implicated. In addition, several other genes exhibit some statistical association with AMD, e.g., age-related maculopathy susceptibility protein 2 (ARMS2) and DNA excision repair protein gene (ERCC6) but more research is needed to establish their significance. Modifiable risk factors for AMD should be discussed with patients whose lifestyle and/or family history place them in an increased risk category. Furthermore, calculation of AMD risk using current models should be recommended as a tool for patient education. It is likely that AMD management in future will be increasingly influenced by assessment of genetic risk as such screening methods become more widely available. © 2013 Spanish General Council of Optometry.

Bone marrow stromal cells stimulate neurite outgrowth over neural proteoglycans (CSPG), myelin associated glycoprotein and Nogo-A

In animal models, transplantation of bone marrow stromal cells (MSC) into the spinal cord following injury enhances axonal regeneration and promotes functional recovery. How these improvements come about is currently unclear. We have examined the interaction of MSC with neurons, using an established in vitro model of nerve growth, in the presence of substrate-bound extracellular molecules that are thought to inhibit axonal regeneration, i.e., neural proteoglycans (CSPG), myelin associated glycoprotein (MAG) and Nogo-A. Each of these molecules repelled neurite outgrowth from dorsal root ganglia (DRG) in a concentration-dependent manner. However, these nerve-inhibitory effects were much reduced in MSC/DRG co-cultures. Video microscopy demonstrated that MSC acted as "cellular bridges" and also "towed" neurites over the nerve-inhibitory substrates. Whereas conditioned medium from MSC cultures stimulated DRG neurite outgrowth over type I collagen, it did not promote outgrowth over CSPG, MAG or Nogo-A. These findings suggest that MSC transplantation may promote axonal regeneration both by stimulating nerve growth via secreted factors and also by reducing the nerve-inhibitory effects of the extracellular molecules present.

Astroglial expression of the P-glycoprotein is controlled by intracellular CNTF

Background - The P-glycoprotein (P-gp), an ATP binding cassette transmembrane transporter, is expressed by astrocytes in the adult brain, and is positively modulated during astrogliosis. In a search for factors involved in this modulation, P-gp overexpression was studied in long-term in vitro astroglial cultures. Results - Surprisingly, most factors that are known to induce astroglial activation in astroglial cultures failed to increase P-gp expression. The only effective proteins were IFNγ and those belonging to the IL-6 family of cytokines (IL-6, LIF, CT-1 and CNTF). As well as P-gp expression, the IL-6 type cytokines - but not IFNγ - stimulated the expression of endogenous CNTF in astrocytes. In order to see whether an increased intracellular level of CNTF was necessary for induction of P-gp overexpression by IL-6 type cytokines, by the same cytokines analysis was carried out on astrocytes obtained from CNTF knockout mice. In these conditions, IFNγ produced increased P-gp expression, but no overexpression of P-gp was observed with either IL-6, LIF or CT-1, pointing to a role of CNTF in the intracellular signalling pathway leading to P-gp overexpression. In agreement with this suggestion, application of exogenous CNTF -which is internalised with its receptor - produced an overexpression of P-gp in CNTF-deficient astrocytes. Conclusions - These results reveal two different pathways regulating P-gp expression and activity in reactive astrocytes, one of which depends upon the intracellular concentration of CNTF. This regulation of P-gp may be one of the long searched for physiological roles of CNTF.

Paediatric drug development:reformulation, in vitro, genomic and in vivo evaluation

Angiotensin converting enzyme (ACE) inhibitors lisinopril and ramipril were selected from EMA/480197/2010 and the potassium-sparing diuretic spironolactone was selected from the NHS specials list for November 2011 drug tariff with the view to produce oral liquid formulations providing dosage forms targeting paediatrics. Lisinopril, ramipril and spironolactone were chosen for their interaction with transporter proteins in the small intestine. Formulation limitations such as poor solubility or pH sensitivity needed consideration. Lisinopril was formulated without extensive development as drug and excipients were water soluble. Ramipril and spironolactone are both insoluble in water and strategies combating this were employed. Ramipril was successfully solubilised using low concentrations of acetic acid in a co-solvent system and also via complexation with hydroxypropyl-β-cyclodextrin. A ramipril suspension was produced to take formulation development in a third direction. Spironolactone dosages were too high for solubilisation techniques to be effective so suspensions were developed. A buffer controlled pH for the sensitive drug whilst a precisely balanced surfactant and suspending agent mix provided excellent physical stability. Characterisation, stability profiling and permeability assessment were performed following formulation development. The formulation process highlighted current shortcomings in techniques for taste assessment of pharmaceutical preparations resulting in early stage research into a novel in vitro cell based assay. The formulations developed in the initial phase of the research were used as model formulations investigating microarray application in an in vitro-in vivo correlation for carrier mediated drug absorption. Caco-2 cells were assessed following transport studies for changes in genetic expression of the ATP-binding cassette and solute carrier transporter superfamilies. Findings of which were compared to in vitro and in vivo permeability findings. It was not possible to ascertain a correlation between in vivo drug absorption and the expression of individual genes or even gene families, however there was a correlation (R2 = 0.9934) between the total number of genes with significantly changed expression levels and the predicted human absorption.

Genomic evaluation during permeability of indomethacin and its solid dispersion

Drug resistance was first identified in cancer cells that express proteins known as multidrug resistance proteins that extrude the therapeutic agents out of the cells resulting in alteration of pharmacokinetics, tissue distribution, and pharmacodynamics of drugs. To this end studies were carried out to investigate the role of pharmacological inhibitors and pharmaceutical excipients with a primary focus on P-glycoprotein (P-gp). The aim of this study was to investigate holistic changes in transporter gene expression during permeability upon formulation of indomethacin as solid dispersion. Initial characterization studies of solid dispersion of indomethacin showed that the drug was dispersed within the carrier in amorphous form. Analysis of permeability data across Caco-2 monolayers revealed that drug absorption increased by 4-fold when reformulated as solid dispersion. The last phase of the work involved investigation of gene expression changes of transporter genes during permeability. The results showed that there were significant differences in the expression of both ATP-binding cassette (ABC) transporter genes as well as solute carrier transporter (SLC) genes suggesting that the inclusion of polyethylene glycol as well as changes in molecular form of drug from crystalline to amorphous have a significant bearing on the expression of transporter network genes resulting in differences in drug permeability. © 2011 Informa UK, Ltd.