Thesis (Ph. D.)--University of Washington, 1997. / Vita. Includes bibliographical references (leaves 171-191).

Thesis (Ph. D.)--University of Rhode Island, 2004. / Typescript. Includes bibliographical references (leaves 104-116).

Thesis (doctoral)--Amsterdam, 1981.

Orientadora: Silvana Martinez Baraldi Artoni / Banca: Laura Satiko Okada Nakaghi / Banca: Daniela Oliveira / Banca: Selma de Fátima Grossi / Banca: Maria Rita Pacheco / Resumo: A muda forçada em poedeiras comerciais tem sido utilizada visando melhorar o desempenho zootécnico das aves por mais um ciclo de produção de ovos. Utilizou-se 32 galinhas poedeiras Hisex Brown com 58 semanas de idade submetidas a diferentes programas de muda forçada para análise do peso e comprimento das diferentes porções intestinais (duodeno, jejuno, íleo, ceco e cólon-reto), com o auxílio de uma balança de precisão e uma fita métrica, respectivamente. As aves foram alojadas em um galpão de postura com gaiolas (2 aves/gaiola) na Unesp, campus de Jaboticabal e expostas à 17 horas de luz diariamente com água e ração à vontade. O delineamento experimental foi inteiramente casualizado com 4 programas contendo 4 aves e 2 coletas aos 28 e 140 dias. Os programas utilizados foram: método Califórnia, baixo nível de cálcio, alto nível de zinco e baixo nível de sódio. Os dados foram submetidos à análise de variância e em caso de diferença significativa, as médias foram comparadas pelo teste de Tukey. Observou-se que aves submetidas ao método Califórnia por 10 dias tiveram respostas biométricas semelhantes aos animais que tiveram alto nível de zinco adicionado à dieta com menor peso corporal e de vísceras, além de menores comprimentos do intestino, além disso, aos 140 dias houve um aumento do peso corpóreo e, também do peso e do comprimento do intestino. / Abstract: The forced molting in commercial laying hens had being utilized for get better the performance of birds for one more cycle of production of eggs. In this study were used 32 Hisex Brown laying hen with 58 weeks of age submitted to different programs of forced molting. This experiment aimed the weight and length of the intestine (duodenum, jejunum, ileum, cecum and rectum). For the weight measurement was utilized one precision scale and for the length was used a measuring tape. The animals were caged in galvanized cage in aviary of Unesp, campus Jaboticabal and submitted of a program of growing light up to 17 hours a day after the induction period and the birds received water and ration ad libitum. The birds were distributed in a randomized experimental assay with 4 programs containing 4 birds and 2 production cycles (28 and 140 days). The animals were distributed into four programs: Califórnia method (control program), diet with low level of calcium, diet with high level of zinc and diet with low level of sodium. The data were submitted to the variance analysis and in case of significant difference, the averages were compared by the test of Tukey. It was observed that birds submitted to the California program were biometric responses similar to the animals that had high level of zinc added to the diet with smaller corporal weight and of visceras. It was observed that smaller lengths of the intestine and increase of corporal weight to the 140 days and increase of the weight and of the length of the intestine, too. / Doutor

Made available in DSpace on 2014-06-11T19:27:15Z (GMT). No. of bitstreams: 0 Previous issue date: 2003Bitstream added on 2014-06-13T18:31:10Z : No. of bitstreams: 1 cerqueira_bf_dr_botfmvz.pdf: 1499209 bytes, checksum: 7ff8a218fa7c9053994488fe2ecdb205 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A oclusão e a reperfusão das artérias esplâncnicas provoca alterações locais e sistêmicas derivadas principalmente da liberação de substâncias citotóxicas e da interação entre neutrófilos e células endoteliais. Buscando-se estudar os efeitos da pentoxifilina e da n-acetilcisteína em lesões provocadas em um modelo de isquemia e reperfusão (I/R), 60 ratos foram divididos em 6 grupos: SAL/CONT (salina/controle): animais submetidos à cirurgia, mas não à I/R, tratados com solução fisiológica; SAL/ISQ (salina/isquemia): animais submetidos à oclusão da artéria celíaca (AC), artéria mesentérica cranial (AMCr) e artéria mesentérica caudal (AMCa) durante 30 minutos, seguidos de 120 minutos de reperfusão, tratados com solução fisiológica; PTX/CONT: animais submetidos à cirurgia, mas não à I/R, tratados com pentoxifilina; PTX/ISQ: animais submetidos à I/R, tratados com pentoxifilina; NAC/CONT: animais submetidos à cirurgia, mas não à I/R, tratados com n-acetilcisteína; NAC/ISQ: animais submetidos à I/R, tratados com n-acetilcisteína. Os parâmetros avaliados foram pressão arterial média (PAM) carótida, pressão venosa jugular, fluxo sangüíneo na aorta e na AMCr, temperatura esofágica, hematócrito, hemogasometria, histologia intestinal, dosagem de espécies reativas ao ácido tiobarbitúrico (TBARS) em duodeno, jejuno e íleo e de malondialdeído (MDA) plasmático e do íleo. Nos grupos submetidos à I/R, a PAM teve queda significativa após 120 minutos de reperfusão; a pressão venosa não mostrou alterações no decorrer do experimento; houve queda do fluxo sangüíneo na aorta após os 30 minutos de isquemia, com queda mais acentuada ao final do período de reperfusão; o fluxo sangüíneo na AMCr diminuiu significativamente após 120 minutos de reperfusão, porém, em menor grau no grupo NAC/ISQ; a freqüência... . / Splanchnic artery occlusion and reperfusion result in local and remote tissue destruction caused by toxic factors released into the circulation and by neutrophil-endothelial cell interactions. In order to evaluate the effects of pentoxifylline (PTX) and n-acetilcysteine (NAC) on ischemia/reperfusion model (I/R), sixty rats were allocated into six groups: 1) SAL/CONT: Sham operation + saline; 2) SALI/ISQ: 30 min celiac, cranial and caudal mesenteric arteries occlusion + 120 min reperfusion + saline; 3) PTX/CONT: Sham operation + PTX (50 mg/kg); 4) PTX/ISQ: I/R + PTX; 5) NAC/SAL: Sham operation + NAC (430 mg/kg) and 6) NAC/ISQ: I/R + NAC. In all groups above mean arterial blood pressure (MABP), vein pressure, aorta and cranial mesenteric artery blood flow, heart rate, esophagus temperature, hematocrit, blood gas determination, histology, thiobarbituric acid reaction species (TBARS) and malondialdehyde (MDA) levels were determined. In I/R groups, MABP decreased significantly 120 minutes after reperfusion (p<0,05); as no difference in vein pressure were observed. Aorta blood flow decreased 30 minutes after ischemia, decreasing dramatically following restoration of blood flow. Cranial mesenteric artery blood flow decreased significantly after reperfusion, although the fall in NAC/ISQ group was attenuated. During the assays, heart rate was reduced, temperature decreased progressively and metabolic acidosis took place. Pentoxiffyline prevented histopathologic signs of injury in duodenum, jejunum and ileum, as well as n-acetilcysteine prevented ileum histopathologic signs of injury. I/R caused TBARS increase in NAC group jejunum and saline group ileum, but no difference was observed in plasmatic and ileal MDA. The present study allowed concluding that PTX was more efficient in preventing histophatologic injury than NAC, but neither PTX nor NAC... (Complete abstract, click electronic address below).

Os diversos estudos com a suplementação de glutamina em situações de estresse fisiológico demonstram o essencial papel deste aminoácido no metabolismo. Agudamente, a suplementação com glutamina aumenta a glutaminemia. Cronicamente, verifica-se maior concentração de glutamina em tecidos, tais como o muscular e o hepático. Entretanto, não é conhecido se esses efeitos são decorrentes diretos da suplementação oral com glutamina ou da redução de sua captação a partir da membrana basolateral de enterócitos. O estudo da cinética na absorção de glutamina traz informações relacionadas à proporção da concentração de glutamina absorvida e a retida no tecido intestinal, de acordo com as doses utilizadas, e aponta quais são as alterações decorrentes da sepse induzida pela endotoxemia. O objetivo deste trabalho foi investigar a cinética de absorção de glutamina em camundongos submetidos à endotoxemia, o que não fora previamente investigado em outros trabalhos. Para avaliar a cinética da absorção intestinal de glutamina foi realizada eversão intestinal em camundongos machos, o que permite a coleta dos líquidos das camadas mucosa e serosa com maior precisão. Foram utilizadas doses de 10, 20, 40 e 50 mM de L-glutamina associada a [14C]-glutamina, nos tempos 0, 5, 10, 20, 30, 40, 60, 90 e 120 minutos. Os resultados obtidos demonstraram que em alta concentração de glutamina (50 mM) ocorre maior absorção em relação à retenção tecidual e esta é realizada por transporte ativo de aminoácidos. Os animais que foram submetidos à endotoxemia por LPS (5mg/kg) apresentaram alterações estruturais no tecido intestinal, detectadas pela histologia. Neste grupo, a retenção tecidual de glutamina foi significativamente maior do que no grupo controle, sobretudo na presença de glicose. Conclui-se que a cinética na absorção de glutamina é dose e tempo dependente em animais saudáveis e que, em condições de endotoxemia, ocorre maior retenção de glutamina no tecido intestinal na presença de glicose. Sugere-se que a via da hexosamina está envolvida; no entanto, mais estudos são necessários para esclarecer tais mecanismos. / The various studies of glutamine supplementation in stressful situations demonstrate the physiological role of this essential amino acid in metabolism. Acutely, supplementation with glutamine improves glutaminemia. Chronically, there is a greater concentration of glutamine in tissues such as muscle and liver. However, it is not known whether these effects are direct result of glutamine oral supplementation or reduced uptake within the basolateral membrane of enterocytes. The kinetic study of the absorption of glutamine provides information related to the ratio of concentration of glutamine absorbed and retained in the intestinal tissue, according to the doses used, and points out the changes resulting from sepsis induced by endotoxemia. The objective of this study was to investigate the kinetics of glutamine uptake in mice subjected to endotoxemia. To evaluate the kinetics of intestinal absorption of glutamine intestinal eversion was performed in male mice, allowing to collect the liquid layers of mucosa and serosa with greater precision. The doses used were 10, 20, 40 and 50 mM L-glutamine associated with [14C]-glutamine at 0, 5, 10, 20, 30, 40, 60, 90 and 120 minutes. The results showed that high concentrations of glutamine (50 mM) a higher absorption occurs in relation to tissue retention and this is accomplished by active transport of amino acids. The animals subjected to endotoxemia by LPS (5mg/kg) showed structural changes in the intestinal tissue, detected by histology. In this group, the tissue retention of glutamine was significantly higher than in the control group, especially in the presence of glucose. It is concluded that the kinetics of glutamine uptake is dose and time dependent in healthy animals, and in conditions of endotoxemia, there is greater retention of glutamine in intestinal tissue in the presence of glucose. It is suggested that the hexosamine pathway is involved; however, more studies are needed to clarify these mechanisms.

Orientador: Nelson Adami Andreollo / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-17T03:26:16Z (GMT). No. of bitstreams: 1 BernisFilho_WalterOctaviano_D.pdf: 14146250 bytes, checksum: 666b2adc648706b4bb83ac6a397a6d6b (MD5) Previous issue date: 2010 / Resumo: Através dos anos muitos fios de sutura foram criados e, depois abandonados, em virtude dos bons resultados obtidos com novos fios. Ainda sim, até hoje não se encontrou um fio cirúrgico totalmente inócuo ao intestino, ou a outros tecidos de um modo geral, tornando sua escolha uma tarefa difícil. Justifica-se, a necessidade de pesquisa de novos materiais no intuito de se encontrar a opção ideal. Neste trabalho foi testado o fio poliglecaprone 25, nas anastomoses do intestino delgado de cão comparando com fios, tradicionalmente usados por outros autores na confecção de anastomoses intestinais, como os fios algodão e poliglactina 910. A cicatrização de anastomoses do intestino delgado foi avaliada, macroscópica e microscópicamente, utilizando três tipos de sutura distintos com os fios poliglecaprone 25, poliglactina 910 e o algodão. Vinte cães machos sem raça definida pesando entre 9 e 16 Kg foram submetidos, após anestesia geral inalatória, a três anastomoses no intestino delgado. A técnica empregada foi a extramucosa com pontos separados e utilizou-se, para cada uma, os fios poliglecaprone 25, a poliglactina 910 e o algodão. Os animais foram separados em 4 grupos de acordo com a avaliação do período pós-operatório: grupoI- 3 dias; grupoII- 7 dias; grupoIII- 14dias; grupo IV- 21dias. Após o período de observação, os animais foram submetidos a eutanásia para coleta de material para análise macroscópica e microscópica. Na avaliação macroscópica os três fios se comportaram bem, com boa coaptação das bordas, porém com moderado grau de aderência entre alças e epiploo, do 3º ao 21º dia do pós-operatório. A avaliação microscópica mostrou inflamação exsudativa com neutrófilos e fibrina que variou de discreta a moderada até o 14º dia; inflamação granulomatosa com presença de macrófagos, células gigantes multinucleadas e células epitelióides mais evidente ao 14º dia para o fio algodão; presença de tecido de granulação (fibroblastos) e fibras colágenas, de forma moderada, a partir do 7º dia para os três fios. Os três tipos de fios de sutura utilizados nesta pesquisa apresentaram comportamento semelhante e podem ser indicados em anastomoses do intestino delgado / Abstract: In this study was evaluate, macroscopically and microscopically, the healing process of intestinal anastomoses in dogs using polyglecaprone 25, polyglactin 910 and cotton sutures. Twenty mongrel dogs, weighting from 9 to 16 Kg were submitted, under general inhalatory anesthesia, to three small intestine anastomoses. The animals were divided into four groups, in accordance with the postoperative observation periods as follows: group I, three days; group II, seven days; group III, fourteen days; group IV, twenty one days. Extramucous technique was used, with those threads, in all four groups. After the observation period the animals were euthanized and samples from the operative site were collected for macroscopic and microscopic evaluations. Macroscopically, all three threads showed good behavior with good coaptation of the edges; however, there occurred a moderate level of adherence between loops of intestine and omentum, from day 3 to day 21, of postoperative period. Microscopically, there was exsudative inflammation, with neutrophils and fibrin, discrete to moderate until day 14. Granulomatous inflammation was also notice accompanied by macrophages, multinucleated giant cells and epithelioid cells, more evident on day 14 in the cotton group. Granulation tissue (fibroblasts) and collagen fibers were also observed, in a moderate pattern, for all three suture materials, from day 7. All three suture threads used in this research showed similar behavior and thus they can be indicated for anastomoses of the small intestine in dogs / Doutorado / Fisiopatologia Cirúrgica / Doutor em Ciências

As doenças do trato digestório nos equinos apresentam altas taxas de morbidade e mortalidade, com diferentes etiologias. Em alguns casos, o emprego da biopsia intestinal se faz necessário para auxílio no diagnóstico dessas enfermidades. No entanto, as técnicas convencionais podem trazer riscos aos pacientes, por serem invasivas, ou não serem elucidativas por apresentarem limitações de acesso a determinados segmentos. O presente estudo teve como objetivo validar uma técnica de biopsia intestinal, intracorpórea, assistida por videolaparoscopia, ainda não descrita na literatura, para coleta de fragmentos de mucosa de jejuno e cólon menor de equinos, que sejam considerados adequados para avaliação histológica. Para tanto, foram utilizados seis equinos machos, da raça Puro Sangue Árabe, com idade de dois anos, sem histórico prévio de doenças do trato digestório, com peso médio de 267 kg. Todos os animais foram submetidos ao mesmo procedimento laparoscópico, instituindo-se apenas jejum alimentar prévio de oito horas. Os equinos foram acompanhados com exame físico e de ultrassonografia abdominal, desde o dia precedente às laparoscopias, até o 15º dia do período pós-operatório, bem como avaliados por meio de hemograma, provas de funções hepática e renal, e análise do líquido peritoneal nos dias 0, 1, 2, 3, 5, 7, 10, 14, 21 e 30. O tempo cirúrgico foi cronometrado, sendo registrado o tempo total, iniciado na criação do primeiro portal de acesso e finalizado ao término da sutura de pele, e os tempos parciais para biopsia de jejuno e cólon menor separadamente, com início na apreensão do segmento intestinal e término quando constatada a polimerização da cola cirúrgica sobre o orifício de acesso da agulha. De cada segmento obtiveram-se dez fragmentos, e posteriormente submetidos à análise histológica. Atribuiu-se escore para cada um deles, sendo considerado 0 fragmentos com qualidade ruim; 1 para qualidade boa e 2 para qualidade ótima. Por sua vez, os considerados viáveis foram somente os que se enquadraram nos escores 1 e 2. Amostras avaliadas como adequadas 11 apresentaram no mínimo 50% dos fragmentos viáveis. A média do tempo total de procedimento foi de 66,50 minutos (± 7,87), enquanto a média do tempo parcial para biopsia de jejuno foi de 14,2 minutos (± 4,3) e a de cólon menor 12,7 minutos (± 5,0). Clinicamente, os animais apresentaram desconforto abdominal nas primeiras 48 horas. Os exames ultrassonográficos do abdômen não revelaram alterações condizentes com peritonite ao longo de todo experimento. Os parâmetros laboratoriais apresentaram apenas características inflamatórias, sendo que o líquido peritoneal permaneceu alterado até o 21º de pós-operatório, havendo normalização de todos os seus valores no 30º dia do estudo. Na inspeção laparoscópica de dois equinos (E2, E4) foi identificada aderência de porção de omento no diafragma. Nas avaliações histológicas de jejuno, uma amostra (E5) de seis foi considerada inadequada, com 5/12 fragmentos viáveis, e em cólon menor, duas (E1, E2) de seis, foram inadequadas, com 4/9 e 5/10 fragmentos viáveis respectivamente. A nova técnica de biopsia intestinal possibilitou a coleta de amostras adequadas de mucosa para análise histológica, de forma segura para os animais, uma vez que as alterações clínicas e laboratoriais foram aquelas relacionadas ao processo inflamatório, compatível com procedimentos laparoscópicos na espécie / Gastrointestinal diseases in horses result in high rates of morbidity and mortality, with different aetiologies. In some cases, an intestinal biopsy is needed to aid in the diagnosis of such diseases. However, the conventional techniques can pose risks to patients for being invasive or for not being elucidating due to having limitations in accessing certain segments. The objective of this study was to validate an intestinal biopsy technique, intracorporeal, assisted by laparoscopy, which has not yet been described in the literature, to collect mucosal fragments from the jejuno and small colon, which might be considered suitable for histological assessment. For such, six male horses were used, Arabian breed, with two years of age, without any records of abdominal diseases, weighing 267 kg in average. All horses were subjected to the same laparoscopic procedure, fasting for eight hours previously to the procedure. All horses were monitored through physical examination and abdominal ultrasonography, from the day previous to laparoscopy, until the 15th postoperative day, as well as hemogram, tests of liver and kidney functions, and analysis of the peritoneal fluid in days 0, 1, 2, 3, 5, 7, 10, 14, 21 and 30. The total laparoscopic procedure time was registered, starting at the moment of the first incision and ending at the moment of the skin closure. The partial times for the jejunal biopsy and small colon biopsy were recorded as well, starting at the grasping of the intestinal segment and ending at the moment of polymerization of the surgical adhesive on the needle access site. From each segment, ten fragments were collected and later subjected to histological analysis. A score was assigned for each one of them, being scored \"0\" fragments of poor quality; \"1\" fragments of good quality and \"2\" fragments of optimal quality. The samples considered viable were only the ones which scored 1 and 2. The samples deemed as adequate showed at least 50% of it fragments to be viable. The average of the surgery total time was of 66,50 minutes (± 7.87), whereas the average of the jejunal biopsy was of 14.2 minutes (± 4.3) and the small colon biopsy time was of 12.7 minutes ( ± 5.0). Clinically, the animals showed mild abdominal 13 discomfort in the first 48 hours. Ultrasonographic examination of the abdomen did not reveal any alterations consistent with peritonitis throughout the entire experiment period. Laboratory parameters presented inflammatory characteristics, and the peritoneal fluid remained altered until the 21th postoperative day, with normalization of all its values on the 30th day of the study. During the laparoscopic inspection of two horses (E2, E4) was identified partial omental adhesion with the diaphragm. In the jejunal histological evaluations, one sample (E5) of six was considered inadequate, with 5/12 viable fragments, and as for the small colon, two (E1, E2) of six were inadequate, with 4/9 and 5/10 viable fragments respectively. The new technique proposed allowed a safe collection of adequate mucosal samples for histological analysis, since clinical and laboratory abnormalities identified were related to the inflammatory process associated to the laparoscopic techniques in horses

Kyoto University (京都大学) / 0048 / 新制・課程博士 / 博士(農学) / 甲第18319号 / 農博第2044号 / 新制||農||1021(附属図書館) / 学位論文||H26||N4826(農学部図書室) / 31177 / 京都大学大学院農学研究科食品生物科学専攻 / (主査)教授 河田 照雄, 教授 伏木 亨, 教授 金本 龍平 / 学位規則第4条第1項該当

Background: Elucidating the role of intestinal drug transporter function in drug development is crucial, as transporter proteins can impact on drug absorption, efficacy and adverse events. In Vitro-In Vivo Extrapolation linked to Physiologically-Based Pharmacokinetic (IVIVE-PBPK) models aim to predict the in vivo impact of transporters from in vitro cell–based transport data and expression-based scaling factors. Currently, these models depend on relative measurements of transporter expression i.e., mRNA or immunoblotting. There is a critical need for physiologically relevant measures of transporter protein abundance to populate these biological frameworks. Objectives: The key objectives were to develop and validate a targeted proteomics workflow to quantify transporter protein abundances in human enterocytes and Caco-2 cells with a QconCAT technique. A cross-laboratory comparison on matched samples was also performed to assess between-laboratory bias in abundance determination. Together with abundance data from each laboratory, BCRP and P-gp transporter activities from Caco-2 cells were used to identify function-abundance relationships, to facilitate the potential development of abundance-function scaling factors. Results: Development of a differential centrifugation technique to obtain plasma membranes was undertaken using MDCK-II and Caco-2 cells. The plasma membrane fraction showed little enrichment from the preceding total membrane fraction and was contaminated with endoplasmic reticulum, as assessed by marker enzyme activities. There were also no differences in Na/K-ATPase, BCRP and P-gp abundances between plasma and total membrane fractions in Caco-2 cells. This may be due to losses of protein from the target membrane fraction, thus, a theoretical framework combining protein assay (BCA) and transporter abundance determinations was proposed. Pilot data on the generation of recovery correction factors using Villin and Na/K-ATPase abundances, to account for protein losses is also presented. The abundances of 6 transporters in jejunal enterocyte membranes (n=3), including the key efflux proteins BCRP (2.56±0.82 fmol/μg), P-gp (1.89±1.07 fmol/μg) and MRP2 (0.59±0.246 fmol/μg) were determined with precision. In addition, peptide losses during protein digestion stages were accounted for in abundance determinations. A cross laboratory comparison of transporter abundances from intestinal (n=4) and Caco-2 cells (n=7) measured in our laboratory and Bertin Pharma (BPh), showed that P-gp abundances were highly correlated (rs=0.72), yet BPh abundances were systematically lower than determined in our laboratory (2.0±2.08 versus. 4.8±3.51 fmol/μg, respectively). No differences or correlations were found for Na/K-ATPase and BCRP abundances between laboratories. A jejunal-Caco-2 cell relative expression factor (REF) for each protein for both laboratories was generated. The P-gp REF was similar for BPh and our laboratory (0.37 vs. 0.4, respectively) however, for BCRP there was a distinct difference (1.11 versus 2.22, respectively). These findings provide the first evidence that employing expression factors generated from abundances quantified in different laboratories may produce altered IVIVE-PBPK outcomes. Functional studies in Caco-2 cells using E-3-S and vinblastine as probes for BCRP and P-gp, respectively, show that protein abundance is more closely correlated to transporter activity than mRNA expression. In addition, it was only possible to verify that increasing P-gp abundances in Caco-2 cells were ranked alongside vinblastine intrinsic clearance, as there was little consistency when estimating Km between the different Caco-2 cell models expressing increasing P-gp abundances, which may be attributed to limited absorptive transport saturation. Thus, forming any conclusions with confidence on concentration dependent abundance-activity relationships was difficult. These data suggest the value of REF scaling factors based on protein abundances, but emphasises the need to generate these from both in vitro and in vivo samples, using the same proteomic workflow. Further work to verify abundance-function relationships is required. Conclusion: A targeted proteomic workflow has been developed allowing quantification of protein abundances for key drug transporters in human gut tissues and cell models. The study has highlighted important areas including losses of targeted proteins, contamination of plasma membrane fractions and standardisation between laboratories that need to be addressed before implementation of transporter abundances into PBPK models is undertaken. Nevertheless, the evidence for a close relationship between transporter abundance and function indicate the potential value of this data for generation of robust REF scaling factors.