Gene expression profile of ABC transporters and cytotoxic effect of ibuprofen and acetaminophen in an epithelial ovarian cancer cell line in vitro

PURPOSES: To determine the basic expression of ABC transporters in an epithelial ovarian cancer cell line, and to investigate whether low concentrations of acetaminophen and ibuprofen inhibited the growth of this cell line in vitro. METHODS: TOV-21 G cells were exposed to different concentrations of acetaminophen (1.5 to 15 μg/mL) and ibuprofen (2.0 to 20 μg/mL) for 24 to 48 hours. The cellular growth was assessed using a cell viability assay. Cellular morphology was determined by fluorescence microscopy. The gene expression profile of ABC transporters was determined by assessing a panel including 42 genes of the ABC transporter superfamily. RESULTS: We observed a significant decrease in TOV-21 G cell growth after exposure to 15 μg/mL of acetaminophen for 24 (p=0.02) and 48 hours (p=0.01), or to 20 μg/mL of ibuprofen for 48 hours (p=0.04). Assessing the morphology of TOV-21 G cells did not reveal evidence of extensive apoptosis. TOV-21 G cells had a reduced expression of the genes ABCA1, ABCC3, ABCC4, ABCD3, ABCD4 and ABCE1 within the ABC transporter superfamily. CONCLUSIONS: This study provides in vitro evidence of inhibitory effects of growth in therapeutic concentrations of acetaminophen and ibuprofen on TOV-21 G cells. Additionally, TOV-21 G cells presented a reduced expression of the ABCA1, ABCC3, ABCC4, ABCD3, ABCD4 and ABCE1 transporters.

Evaluation of the role of ATP-binding cassette transporters as a defence mechanism against temephos in populations of Aedes aegypti

The role of ATP-binding cassette (ABC) transporters in the efflux of the insecticide, temephos, was assessed in the larvae of Aedes aegypti. Bioassays were conducted using mosquito populations that were either susceptible or resistant to temephos by exposure to insecticide alone or in combination with sublethal doses of the ABC transporter inhibitor, verapamil (30, 35 and 40 μM). The best result in the series was obtained with the addition of verapamil (40 μM), which led to a 2x increase in the toxicity of temephos, suggesting that ABC transporters may be partially involved in conferring resistance to the populations evaluated.

P-glycoprotein-mediated resistance to chemotherapy in cancer cells: using recombinant cytosolic domains to establish structure-function relationships

Resistance to chemotherapy in cancer cells is mainly mediated by overexpression of P-glycoprotein (Pgp), a plasma membrane ATP-binding cassette (ABC) transporter which extrudes cytotoxic drugs at the expense of ATP hydrolysis. Pgp consists of two homologous halves each containing a transmembrane domain and a cytosolic nucleotide-binding domain (NBD) which contains two consensus Walker motifs, A and B, involved in ATP binding and hydrolysis. The protein also contains an S signature characteristic of ABC transporters. The molecular mechanism of Pgp-mediated drug transport is not known. Since the transporter has an extraordinarily broad substrate specificity, its cellular function has been described as a "hydrophobic vacuum cleaner". The limited knowledge about the mechanism of Pgp, partly due to the lack of a high-resolution structure, is well reflected in the failure to efficiently inhibit its activity in cancer cells and thus to reverse multidrug resistance (MDR). In contrast to the difficulties encountered when studying the full-length Pgp, the recombinant NBDs can be obtained in large amounts as soluble proteins. The biochemical and biophysical characterization of recombinant NBDs is shown here to provide a suitable alternative route to establish structure-function relationships. NBDs were shown to bind ATP and analogues as well as potent modulators of MDR, such as hydrophobic steroids, at a region close to the ATP site. Interestingly, flavonoids also bind to NBDs with high affinity. Their binding site partly overlaps both the ATP-binding site and the steroid-interacting region. Therefore flavonoids constitute a new promising class of bifunctional modulators of Pgp.

A novel ABCG-like transporter of Trypanosoma cruziis involved in natural resistance to benznidazole

Benznidazole (BZ) is one of the two drugs used for Chagas disease treatment. Nevertheless therapeutic failures of BZ have been reported, which were mostly attributed to variable drug susceptibility among Trypanosoma cruzistrains. ATP-binding cassette (ABC) transporters are involved in a variety of translocation processes and some members have been implicated in drug resistance. Here we report the characterisation of the first T. cruzi ABCG transporter gene, named TcABCG1, which is over-expressed in parasite strains naturally resistant to BZ. Comparison ofTcABCG1 gene sequence of two TcI BZ-resistant strains with CL Brener BZ-susceptible strain showed several single nucleotide polymorphisms, which determined 11 amino acid changes. CL Brener transfected with TcI transporter genes showed 40-47% increased resistance to BZ, whereas no statistical significant increment in drug resistance was observed when CL Brener was transfected with the homologous gene. Only in the parasites transfected with TcI genes there was 2-2.6-fold increased abundance of TcABCG1transporter protein. The analysis in wild type strains also suggests that the level of TcABCG1transporter is related to BZ natural resistance. The characteristics of untranslated regions of TcABCG1genes of BZ-susceptible and resistant strains were investigated by computational tools.

O subsistema americano, Rio Branco e o ABC

O artigo discute as motivações e objetivos que levaram o Brasil a participar de negociações com a Argentina e o Chile, entre 1907 e 1915, para a assinatura de um tratado de "cordial inteligência política", conhecido como ABC. Com vistas a situar essa iniciativa no contexto internacional do período, examina a configuração do subsistema americano bem como o papel do pan-americanismo. A fim de identificar seu significado político, analisa os fundamentos da política externa de Rio Branco (1902-1912) e as mais relevantes interpretações do ABC na historiografia, utilizando como recursos metodológicos os conceitos de sistemas de Estados e de hegemonia.

Um modelo de reconciliação de dados para o custeio baseado-em-atividades (ABC)

A mudança na estrutura dos custos, em função de novas tecnologias de produção, de uma maior diversificação imposta pela competitividade empresarial e da redução dos custos de medição, criou condições para o aparecimento de técnicas de gerenciamento de custos voltadas para a captação mais precisa das despesas indiretas que cada produto consome. Nesse mister destaca-se o Custeio Baseadoem- Atividades/Gerenciamento Baseado-em-Atividades (ABC/ABM) como uma das técnicas mais importantes surgidas nos últimos anos, que proporciona uma estimativa mais precisa das despesas indiretas reais que incidem em cada produto. Existem, contudo, certos custos cuja obtenção é difícil mesmo com a utilização de técnicas como o ABC. Isso se deve não só pela impossibilidade, em alguns casos, de serem determinados com adequada exatidão, como, em outros, pelo fato de o investimento na sua medição ultrapassar o benefício a ser obtido com ele. Nesse sentido, o presente trabalho desenvolve, para o fluxo de custos de uma empresa, inspirado na reconciliação de dados utilizada nos processos das plantas químicas, um modelo para a determinação de todos os custos indiretos, partindo do conhecimento preciso de alguns dos custos que compõem o citado fluxo de custos. O modelo desenvolvido utiliza programação matemática não-linear.

Controvérsias acerca do sistema de custos ABC

Em vista do modismo em analisar e implantar nas empresas o Sistema de Custos ABC, reforçado pelas organizações internacionais de consultoria, o artigo traz a controvérsia que persiste na literatura especializada. A maior acurácia embutida no custeio ABC, principal premissa dos seus proponentes, é questionada por seus opositores alegando que nem sempre leva às melhores decisões empresariais. Pesquisa sobre utilização de sistemas de custos nas empresas mostra seu uso não generalizado e sinaliza sua maior adequação em situações específicas.

The 245 kb amplified chromosome of Leishmania (V.) braziliensis contains a biopterin transporter gene

Leishmania (V.) braziliensis M2903 presents a small linear and stable 245 kb chromosome originating from a genomic amplification. Similar amplifications present in other species of Leishmania contain a gene coding for a biopterin transporter. Since Leishmania is auxotrophic for this metabolite, this amplification could result from the need to better capture biotpterin from growth media under specific circumstances. In this paper we show that this gene is also present in L. (V.) braziliensis small chromosome, which shares sequences with other genomic amplifications already described.