Recapitulating the Pharmacological Interactions of Cetuximab with Sunitinib and Cisplatin in Head and Neck Carcinoma Cells in vitro.

INTRODUCTION: Cisplatin is extensively used in the treatment of head and neck carcinomas. Cetuximab combination therapy is employed in recurrent and metastatic settings. Sunitinib showed positive results in the treatment of head and neck carcinomas, both as monotherapy or in combination with cetuximab. Nonetheless, the mechanism governing these pharmacological interactions is largely unresolved. This study investigates the impact of cetuximab on the cytotoxicity of cisplatin and sunitinib using cells representative of head and neck carcinoma and the oral epithelium. METHODS: The uptake and efflux activities of cells were determined using the prototypical fluorescent substrates 4-[4-[dimethylamino]styryl)-1-methyl pyridinium iodide, Hoechst 33342, and calcein-AM in the presence or absence of specific inhibitors in cells pretreated with cetuximab. The expression of key uptake and efflux drug transporters was analyzed using qPCR and immunofluorescence. Cisplatin and sunitinib cytotoxicities after cetuximab pretreatment were evaluated using the PrestoBlue viability assay. RESULTS: Both tumor and nontumor cells showed significant active drug transport activity. Cetuximab substantially deregulated the expression of key transporters involved in drug resistance in head and neck cancer cells. Transporter expression in the nontumor cell was unaffected. Upon cetuximab pretreatment, the half maximal effective toxic concentration of cisplatin was reduced by 0.75-fold and sunitinib by 0.82-fold in cancer cells. Nontumor cells were not sensitive to cisplatin or sunitinib under the conditions tested. CONCLUSION: Cetuximab regulates the expression and activity of key membrane drug transporters in head and neck cancer cells, involved in drug resistance. The deregulation of the transport mechanism behind cisplatin and sunitinib uptake reverses drug resistance and enhances the cytotoxicity of both drugs.

In Vitro Characterization of Renal Drug Transporter Activity in Kidney Cancer.

The activity of drug transporters is central to the secretory function of the kidneys and a defining feature of renal proximal tubule epithelial cells (RPTECs). The expression, regulation, and function of these membrane-bound proteins is well understood under normal renal physiological conditions. However, the impact of drug transporters on the pathophysiology of kidney cancer is still elusive. In the present study, we employed different renal cell carcinoma (RCC) cell lines and a prototypical non-malignant RPTEC cell line to characterize the activity, expression, and potential regulatory mechanisms of relevant renal drug transporters in RCC in vitro. An analysis of the uptake and efflux activity, the expression of drug transporters, and the evaluation of cisplatin cytotoxicity under the effects of methylation or epidermal growth factor receptor (EGFR) inhibition showed that the RCC cells retained substantial drug transport activity. In RCC cells, P-glycoprotein was localized in the nucleus and its pharmacological inhibition enhanced cisplatin toxicity in non-malignant RPTECs. On the other hand, methylation inhibition enhanced cisplatin toxicity by upregulating the organic cation uptake activity in RCC cells. Differential effects of methylation and EGFR were observed in transporter expression, showing regulatory heterogeneity in these cells. Interestingly, the non-malignant RPTEC cell line that was used lacked the machinery responsible for organic cation transport, which reiterates the functional losses that renal cells undergo in vitro.

Microfluidic-based prostate cancer model for investigating the secretion of prostate-specific antigen and microRNAs in vitro.

The study of prostate cancer in vitro relies on established cell lines that lack important physiological characteristics, such as proper polarization and expression of relevant biomarkers. Microphysiological systems (MPS) can replicate cancer microenvironments and lead to cellular phenotypic changes that better represent organ physiology in vitro. In this study, we developed an MPS model comprising conventional prostate cancer cells to evaluate their activity under dynamic culture conditions. Androgen-sensitive (LNCaP) and androgen-insensitive (PC3) cells were grown in conventional and 3D cultures, both static and dynamic. Cell morphology, the secretion of prostate-specific antigen, and the expression of key prostate markers and microRNAs were analyzed. LNCaP formed spheroids in 3D and MPS cultures, with morphological changes supported by the upregulation of cytokeratins and adhesion proteins. LNCaP also maintained a constant prostate-specific antigen secretion in MPS. PC3 cells did not develop complex structures in 3D and MPS cultures. PSA expression at the gene level was downregulated in LNCaP-MPS and considerably upregulated in PC3-MPS. MicroRNA expression was altered by the 3D static and dynamic culture, both intra- and extracellularly. MicroRNAs associated with prostate cancer progression were mostly upregulated in LNCaP-MPS. Overall dynamic cell culture substantially altered the morphology and expression of LNCaP cells, arguably augmenting their prostate cancer phenotype. This novel approach demonstrates that microRNA expression in prostate cancer cells is sensitive to external stimuli and that MPS can effectively promote important physiological changes in conventional prostate cancer models.

Intervenção coronária percutânea em pontes de veia safena com uso de stents farmacológicos: resultados agudos e tardios dos pacientes incluídos no registro DESIRE / Percutaneous coronary intervention en saphenous vein bypass grafts with drug-eluting stents: early and late outcomes of patients in the DESIRE registry

Mesmo com o advento dos stents farmacológicos, a intervenção coronária percutânea em pontes de veia safena permanece desafiadora, com elevada incidência de complicações agudas e incerteza dos resultados clínicos de pacientes com lesões em pontes de veia safena tratados com stents farmacológicos. Método: Entre maio de 2002 e janeiro de 2009, 151 pacientes com 196 lesões em pontes de veia safena foram consecutivamente submetidos a intervenção coronária percutânea com 211 stents farmacológicos e incluídos neste estudo. Os pacientes foram pré-tratados com enoxaparina, clopidogrel e ácido acetilsalicílico por 3 a 5 dias antes da intervenção coronária percutânea. A terapia antiplaquetária dupla (ácido acetilsalicílico + clopidogrel) foi prescrita por 12 meses. Objetivou-se determinar as taxas de eventos cardíacos adversos maiores (ECAM) no período hospitalar e na fase tardia. Seguimento clínicofoi obtido com 1, 6 e 12 meses e, então, anualmente...

Introduction: Despite the advent of drug-eluting stents, percutaneous coronary interventions in saphenous vein bypass grafts remain a challenge, with a high incidence of early complications and uncertainty regarding late outcomes. This study was aimed at evaluating clinical outcomes of patients with saphenous vein graft lesions treated with drug-eluting stents. Method: From May 2002 to January 2009, a total of 151 patients with 196 saphenous vein graft lesions were consecutively submitted to percutaneous coronary intervention with 211 drug-eluting stents and included in this trial. Patients were pretreated with enoxiparin, clopidogrel and acetylsalicylic acid for 3 to 5 days prior to percutaneous coronary intervention. Dual antiplatelet therapy (acetylsalicylic acid + clopidogrel) was prescribed for 12 months. The objective was to determine the rate of major adverse cardiac events (MACE) during hospitalization and in the long-term. Clinical follow-up was obtained at 1, 6 and 12 months and then, yearly after that...

Implante de stent farmacológico para o tratamento da reestenose de outro stent farmacológico: análise tardia do registro DESIRE / Drug-Eluting Stent Implantation for the Treatment of Drug-Eluting Stent Restenosis: Long-Term Analysis of the DESIRE Registry

Fundamentos: Apesar da notória redução nas taxas de reestenose (RIS) com os stents farmacológicos (SF), essa complicação ainda ocorre em 5 por cento a 25 por cento dos casos. A história natural e o melhor tratamento para a reestenose de SF ainda não estão definidos. Método: Entre maio de 2002 e novembro de 2007, todos os pacientes com RIS de SF tratados com outro SF foram consecutivamente incluídos neste registro. O tipo de SF implantado para tratar a RIS ficou a critério do operador. Acompanhamento clínico foi obtido com 1 , 6 e 12 meses e, então, anualmente. O objetivo primário foi avaliar a incidência acumulada de eventos cardíacos maiores (ECAM). Resultados: Dos 45 pacientes (53 lesões) tratados percutaneamente com outro SF, 66,6 por cento eram do sexo masculino. Diabetes melito foi observado em 35,5 por cento desses pacientes. A maioria das reestenosis foi do tipo focal (75 por cento). O diâmetro de referência do vaso tratado e a extensão da lesão foram de 2,99 + - 0,5 mm e 11,33 + - 5,8 mm, respectivamente. Em 28 por cento dos casos optou-se por implantar um SF diferente daquele que apresentou RIS. Sucesso angiográfico foi obtido em 100 por cento dos casos. Seguimento clínico foi obtido em toda a população...

Background: Despite the marked reduction in restenosis rates following drug-eluting stent (DES) use, this complication still happens in 5%-25% of the cases. The natural history and the best treatment for DES in-stent restenosis are still to be defined. Methods: Between May 2002 and November 2007, all consecutive cases of DES restenosis treated with another DES were included in this registry. The type of DES to be deployed was at the surgeon's discretion. Follow-up data were obtained at 1, 6, and 12 months and then annually. The primary endpoint of this study was the incidence of combined major cardiac events (MACE). Results: Of the 45 patients (53 lesions) submitted to new percutaneous coronary intervention (PCI) with DES, 66.6% were men. Diabetes mellitus was observed in 35.5% of the patients. Most in-stent restenosis were focal (75%). Reference vessel diameter and lesion length were 2.99 ± 0.5 mm and 11.33 ± 5.8 mm, respectively. A different type of DES was preferred in 28% of the cases. Angiographic success was achieved in all cases. Clinical data of all population were obtained (mean 2.7 ± 1.1 years). In the long term, the cumulative rate of MACE was 13.3%, and 5 (11.1%) patients manifested recurrent symptoms. Conclusions: The use of a DES to treat a previous DES in-stent restenosis represents a safe and efficient approach, with low rates of adverse events in the short and long terms.

Ciclo de vida familiar / Family life-cycle

Investigou-se a associação entre o conhecimento do processo emocional de transição em cada etapa do ciclo vital familiar (novo casal, família com filhos pequenos, família com filhos adolescentes, família com filhos adultos, família no estágio tardio da vida) com as mudanças necessárias para prosseguir no desenvolvimento familiar. A amostra foi de 323 universitários, casados, sendo 215 do sexo feminino e 108 do sexo masculino. Foram construídos três questionários, totalizando 65 itens, para se coletar os dados da amostra. Para análise dos dados utilizou-se o teste de correlação D de Somer; processado pelo programa SPSS (Statistic Package for the Social Sciences). Verificou-se que há correlação das respostas referentes às tarefas das diferentes etapas do ciclo de vida familiar com o processo emocional de transição. Observou-se, no entanto, que o número de concordâncias significativas aumentaram ao longo das etapas do ciclo de vida familiar; começando com poucas correlações entre os títulos dos questionários nas etapas do ciclo de vida familiar até chegar a uma correlação de todos os itens avaliados na última etapa