Enhancement of viability of radiosensitive (PBMC) and resistant (MDA-MB-231) clones in low-dose-rate cobalt-60 radiation therapy

AbstractObjective:In the present study, the authors investigated the in vitrobehavior of radio-resistant breast adenocarcinoma (MDA-MB-231) cells line and radiosensitive peripheral blood mononuclear cells (PBMC), as a function of different radiation doses, dose rates and postirradiation time kinetics, with a view to the interest of clinical radiotherapy.Materials and Methods:The cells were irradiated with Co-60, at 2 and 10 Gy and two different exposure rates, 339.56 cGy.min–1 and the other corresponding to one fourth of the standard dose rates, present over a 10-year period of cobalt therapy. Post-irradiation sampling was performed at pre-established kinetics of 24, 48 and 72 hours. The optical density response in viability assay was evaluated and a morphological analysis was performed.Results:Radiosensitive PBMC showed decrease in viability at 2 Gy, and a more significant decrease at 10 Gy for both dose rates. MDAMB- 231 cells presented viability decrease only at higher dose and dose rate. The results showed MDA-MB-231 clone expansion at low dose rate after 48–72 hours post-radiation.Conclusion:Low dose rate shows a possible potential clinical impact involving decrease in management of radio-resistant and radiosensitive tumor cell lines in cobalt therapy for breast cancer.

Involvement of miR-30c in resistance to doxorubicin by regulating YWHAZ in breast cancer cells

MicroRNAs (miRNAs) are small RNA molecules that modulate gene expression implicated in cancer, which play crucial roles in diverse biological processes, such as development, differentiation, apoptosis, and proliferation. The aim of this study was to investigate whether miR-30c mediated the resistance of breast cancer cells to the chemotherapeutic agent doxorubicin (ADR) by targeting tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta (YWHAZ). miR-30c was downregulated in the doxorubicin-resistant human breast cancer cell lines MCF-7/ADR and MDA-MB-231/ADR compared with their parental MCF-7 and MDA-MB-231 cell lines, respectively. Furthermore, we observed that transfection of an miR-30c mimic significantly suppressed the ability of MCF-7/ADR to resist doxorubicin. Moreover, the anti-apoptotic gene YWHAZ was confirmed as a target of miR-30c by luciferase reporter assay, and further studies indicated that the mechanism for miR-30c on the sensitivity of breast cancer cells involved YWHAZ and its downstream p38 mitogen-activated protein kinase (p38MAPK) pathway. Together, our findings provided evidence that miR-30c was one of the important miRNAs in doxorubicin resistance by regulating YWHAZ in the breast cancer cell line MCF-7/ADR.

Synthesis of geranylhydroquinone derivatives with potencial cytotoxic activity

Natural geranylhydroquinone 1 and geranyl-p-methoxyphenol 2 were prepared by Electrophilic Aromatic Substitution (EAS) reactions between geraniol and 1,4-hydroquinone or p-methoxyphenol respectively, using BF3∙Et2O as a catalyst. Furthermore, natural geranylquinone 3, geranyl-1,4-dimethoxyquinone 4 and the new geranyl-4-methoxyphenyl acetate 5 were obtained by chemical transformations of 1 and 2. The compounds were evaluated for their in vitro cytotoxicity activities against cultured human cancer cells of PC-3 human prostate cancer, MCF-7 and MDA-MB-231 breast carcinoma, and Dermal Human Fibroblasts DHF. IC50 values were in the µM range.

RNAi-mediated knockdown of FANCF suppresses cell proliferation, migration, invasion, and drug resistance potential of breast cancer cells

Fanconi anemia complementation group F protein (FANCF) is a key factor, which maintains the function of FA/BRCA, a DNA damage response pathway. However, the functional role of FANCF in breast cancer has not been elucidated. We performed a specific FANCF-shRNA knockdown of endogenous FANCF in vitro. Cell viability was measured with a CCK-8 assay. DNA damage was assessed with an alkaline comet assay. Apoptosis, cell cycle, and drug accumulation were measured by flow cytometry. The expression levels of protein were determined by Western blot using specific antibodies. Based on these results, we used cell migration and invasion assays to demonstrate a crucial role for FANCF in those processes. FANCF shRNA effectively inhibited expression of FANCF. We found that proliferation of FANCF knockdown breast cancer cells (MCF-7 and MDA-MB-435S) was significantly inhibited, with cell cycle arrest in the S phase, induction of apoptosis, and DNA fragmentation. Inhibition of FANCF also resulted in decreased cell migration and invasion. In addition, FANCF knockdown enhanced sensitivity to doxorubicin in breast cancer cells. These results suggest that FANCF may be a potential target for molecular, therapeutic intervention in breast cancer.

Effect of penehyclidine hydrochloride on β-arrestin-1 expression in lipopolysaccharide-induced human pulmonary microvascular endothelial cells

β-arrestins are expressed proteins that were first described, and are well-known, as negative regulators of G protein-coupled receptor signaling. Penehyclidine hydrochloride (PHC) is a new anti-cholinergic drug that can inhibit biomembrane lipid peroxidation, and decrease cytokines and oxyradicals. However, to date, no reports on the effects of PHC on β-arrestin-1 in cells have been published. The aim of this study was to investigate the effect of PHC on β-arrestin-1 expression in lipopolysaccharide (LPS)-induced human pulmonary microvascular endothelial cells (HPMEC). Cultured HPMEC were pretreated with PHC, followed by LPS treatment. Muscarinic receptor mRNAs were assayed by real-time quantitative PCR. Cell viability was assayed by the methyl thiazolyl tetrazolium (MTT) conversion test. The dose and time effects of PHC on β-arrestin-1 expression in LPS-induced HPMEC were determined by Western blot analysis. Cell malondialdehyde (MDA) level and superoxide dismutase (SOD) activity were measured. It was found that the M3 receptor was the one most highly expressed, and was activated 5 min after LPS challenge. Furthermore, 2 μg/mL PHC significantly upregulated expression of β-arrestin-1 within 10 to 15 min. Compared with the control group, MDA levels in cells were remarkably increased and SOD activities were significantly decreased in LPS pretreated cells, while PHC markedly decreased MDA levels and increased SOD activities. We conclude that PHC attenuated ROS injury by upregulating β-arrestin-1 expression, thereby implicating a mechanism by which PHC may exert its protective effects against LPS-induced pulmonary microvascular endothelial cell injury.

Constituintes químicos de Parmotrema lichexanthonicum Eliasaro & Adler: isolamento, modificações estruturais e avaliação das atividades antibiótica e citotóxica

From the lichen Parmotrema lichexantonicum were isolated the depsidone salazinic acid, the xanthone lichexanthone, and the depside atranorin. The two major compounds, salazinic acid and lichexanthone, were selected for structure modifications. Salazinic acid afforded O-alkyl salazinic acids, some of them potentially cytotoxic against tumor cell lines (HCT-8, SF-295 and MDA/ MB - 435). From lichexanthone were obtained norlichexanthone, 3-O-methylnorlichexanthone, 3-O-methyl-6-O-prenylnorlichexanthone, 3,6-di-O-prenyl-norlichexanthone, 3,6-bis[(3,3-dimethyloxyran-2-il)methoxy]-1-hydroxy-8-methyl-9H-xanten-9-one and 3,6-bis[3-(dimethylamine)propoxy]-1-hydroxy-8-methyl-9H-xanten-9-one. The last compound was the most active against S. aureus.

Trachylobane and kaurane diterpenes from Croton floribundus spreng

A new trachylobane diterpene ent-trachyloban-18,19-diol (1) was isolated from root bark of Croton floribundus, along with known diterpenes ent-trachyloban-19-oic acid (2), 15b-hydroxy-ent-trachyloban-19-oic acid (3), ent-trachyloban-19-ol (4), ent-kaur-16-en-19-oic acid (5), ent-kaur-16-ene-6a,19-diol (6) and ent-16a-hydroxykaur-11-en-19-oic acid (7). ent-trachyloban-18,19-diol (1) was submitted to derivatization reactions affording four new compounds (8-11). Cytotoxic activity of diterpenes 1, 3, 4, 7-11 against three human cancer cell lines was evaluated. No compounds showed cytotoxic potential with IC50 values greater than 25 mg/mL. Compound 6 was evaluated against five human cancer cell lines, showing moderate effect against three cancer cell lines, MDA-MB-435, HCT-8 and HCT-116, with IC50 values of 14.32, 13.47 and 12.1 mg/mL, respectively.

Mycobacterium leprae downregulates the expression of PHEX in Schwann cells and osteoblasts

Neuropathy and bone deformities, lifelong sequelae of leprosy that persist after treatment, result in significant impairment to patients and compromise their social rehabilitation. Phosphate-regulating gene with homologies to endopeptidase on the X chromosome (PHEX) is a Zn-metalloendopeptidase, which is abundantly expressed in osteoblasts and many other cell types, such as Schwann cells, and has been implicated in phosphate metabolism and X-linked rickets. Here, we demonstrate that Mycobacterium leprae stimulation downregulates PHEX transcription and protein expression in a human schwannoma cell line (ST88-14) and human osteoblast lineage. Modulation of PHEX expression was observed to a lesser extent in cells stimulated with other species of mycobacteria, but was not observed in cultures treated with latex beads or with the facultative intracellular bacterium Salmonella typhimurium. Direct downregulation of PHEX by M. leprae could be involved in the bone resorption observed in leprosy patients. This is the first report to describe PHEX modulation by an infectious agent.

Synergism between dipyridamole and cisplatin in human breast cancer cells in vitro

Cisplatin is very effective in the treatment of metastatic breast cancer. However, the development of cellular resistance is a serious problem in cisplatin chemotherapy. In the present work, the effects of dipyridamole (DPM) on the cellular accumulation and cytotoxicity of cisplatin was studied in cisplatinsensitive (MDA/S) and cisplatinresistant (MDA/R) human breast cancer cells. In the presence of 30 µM DPM, the IC50 of cisplatin was reduced by 39% for both cell lines. Combination index analysis revealed that cisplatin and dipyridamole interact synergistically in MDA/R cells. In the MDA/S cells, the cellular accumulation of cisplatin increased by 57 ± 8% in the presence of 30 µM DPM. In the MDA/R cells, the cellular accumulation of cisplatin remained the same with or without 30 µM DPM. The results suggest that the enhancement of cisplatin cytotoxicity by DPM in MDA/S cells may be related to a DPM-induced increase in cisplatin accumulation, but the enhanced cytotoxicity in MDA/R cells employs a mechanism that does not involve an increase in the cellular accumulation of cisplatin.

Serum dosage of CPK-MB in dogs with ST deviation by chemiluminescence

Abstract: Although frequently in humans, hypoxic and ischemic heart diseases are poorly documented in dogs, with only few reports of acute myocardial infarction (AMI) in this species. Some electrocardiographic findings might suggest myocardium hypoxia/ischemia, like ST segment elevation or depression, but there are no studies showing whether deviations in ST segment are associated to myocardial injury and serum increase of creatine phosphokinase (CPK-MB). In order to investigate possible myocardial cells injury in poor perfusion conditions, 38 dogs were studied, 20 with normal electrocardiogram and 18 with ST segment elevation or depression, recorded in lead II, at a paper speed of 50 mm/sec and N sensibility (1mV=1cm). Serum measurement of creatine phosphokinase isoenzyme MB (CPK-MB) in normal dogs (group 1) determined control values (in ng/mL), which were compared to those obtained from dogs with deviation (group 2), which allowed confirmation or not of myocardial injury. CPK-MB mean values obtained from dogs in groups 1 and 2 were 0.540ng/ml (SD±0.890)ng/mL and 0.440ng/mL (SD±1.106), respectively. At a significance level of 5%, the relation of CPK-MB with age, mass and total creatine phosphokinase (CPK-T) was not significant in groups 1 and 2. CPK-MB showed no difference, at 5% level, between groups 1 and 2. In conclusion, it is possible to use the human chemiluminescent immunometric assay kit in canine species and that hypoxia/ischemia revealed by ST segment deviation does not mean significant myocardium injury.

Aceitação sensorial de híbridos de milho doce e híbridos de milho verde em intervalos de colheita

A composição química e a textura dos grãos do milho doce estão, diretamente, relacionadas com as classes de endosperma que interferem na aceitação do produto pelos consumidores. Por isso, este trabalho teve por objetivo avaliar as características sensoriais de híbridos de milho doce e de híbridos de milho verde, em função de intervalos de colheita. O experimento foi instalado na Estação Experimental de Pesquisa da Syngenta Seeds Ltda, no município de Uberlândia-MG. O delineamento experimental foi de blocos casualizados, em esquema fatorial 8x4, correspondente a oito híbridos (seis híbridos experimentais de milho doce - SWC03, SWC04, SWC05, SWC06, SWC07 e SWC08 e dois híbridos de milho verde SWC01 e SWC02) e quatro intervalos de colheita (26, 28, 30 e 32 dias após a colheita - DAF), com três repetições. No estudo, que envolveu 41 pessoas (25 mulheres e 16 homens), utilizando-se o teste de aceitação com escala hedônica, concluiu-se que os híbridos de milho doce têm maior aceitação que os híbridos de milho verde, independentemente do intervalo de colheita. Colheitas tardias (32 DAF) refletem-se em redução da aceitabilidade do milho doce, que se assemelha à do milho verde (entre indiferente e gostei ligeiramente, para cor, e desgostei ligeiramente e desgostei moderadamente, para textura e sabor). Para todos os híbridos testados, a colheita, quando realizada entre 26 e 28 DAF, proporciona produtos com melhores resultados sensoriais (textura, sabor e cor).

Custo da produção de silagens em sistemas de integração lavoura-pecuária sob plantio direto

Objetivou-se, com este trabalho, avaliar o custo operacional total (COT) da produção de silagens em sistemas produtivos de Integração Lavoura-Pecuária (ILP), a produtividade de massa seca (PMS) e a quantidade de proteína bruta (PB) e nutrientes digestíveis totais (NDT), por área, em cada modalidade de cultivo sob condições irrigadas, no Cerrado, durante dois anos agrícolas (2010/2011 e 2011/2012). O delineamento experimental foi o de blocos casualizados, com quatro repetições, sendo os tratamentos constituídos de três modalidades de cultivo das culturas do milho e do sorgo para silagem (solteiro; em consórcio com a Brachiaria brizantha cv. Xaraés e com Panicum maximum cv. Tanzânia). Constatou-se que a silagem de sorgo apresentou custo de produção inferior ao da silagem de milho. Os maiores COTs foram obtidos no consórcio de milho com capim Xaraés (R$ 2.046,93 e de R$ 2.385,40) e, os menores, nos cultivos de sorgo solteiro (R$ 1.815,93 e de R$ 1.941,62), para os dois anos agrícolas. Os tratamentos com cultivos exclusivos (de sorgo e de milho) proporcionaram maiores valores de PMS no primeiro ano; porém, no segundo ano, os maiores valores foram proporcionados pelos cultivos consorciados de ambas as espécies com Panicum maximum cv. Tanzânia. Os valores de PB e NDT por área variaram entre os tratamentos nos dois anos agrícolas e seguiram o padrão da PMS, sendo os maiores valores proporcionados também pelos consórcios de sorgo e milho com o capim Tanzânia.

Calibração de sondas capacitivas, funções físico-hídricas e variação do armazenamento de água em um argissolo cultivado com palma forrageira

Objetivou-se, com este trabalho, realizar a calibração de três sondas capacitivas, visando a utilizá-las na definição de funções físico-hídricas e na análise da variação espacial e temporal do conteúdo (q) e do armazenamento de água, em um Argissolo Vermelho-Amarelo, cultivado com três clones de palma forrageira, no Semiárido pernambucano. O estudo foi executado numa área experimental situada em Serra Talhada, PE. Para isso, foi realizada a calibração de três sondas capacitivas, usando-se dados de densidade aparente do solo, de umidade gravimétrica e de frequências relativas, em cinco condições de umidade do solo. Aplicou-se o método do perfil instantâneo, para determinação do fluxo vertical de água, e do potencial matricial de água no solo. Em seguida, analisou-se a variação espaço-temporal do q e do armazenamento de água no solo. As medidas de frequência relativa das sondas capacitivas apresentaram elevada relação com os dados de q (R2 > 0,87). A partir do método do perfil instantâneo, foi possível determinar as funções da condutividade hidráulica e do potencial total de água no solo, em relação ao q, com elevados coeficientes de determinação (R2 > 0,91). Não houve diferenças significativas do q entre os clones de palma forrageira.

Atratividade de adultos de Chrysoperla externa (Hagen, 1861) aos compostos voláteis de coentro, endro e erva-doce (Apiaceae) em condições de laboratório

Espécies de Apiaceae dispõem de óleos essenciais, nos quais podem ocorrer compostos voláteis, que funcionam como sinais para atração e manutenção de inimigos naturais nas áreas cultivadas. Com base nestas características, este trabalho objetivou avaliar a atratividade aos adultos do predador Chrysoperla externa. Foram utilizados folhas e caules de coentro, endro e erva-doce, coletados aos 30 e 60 dias após a semeadura. As plantas foram dispostas em olfatômetro de quatro vias (formato de "X") disponibilizando-se os odores para machos e fêmeas, virgens e acasalados, em testes de livre escolha. Ao serem liberados individualmente no interior do olfatômetro, foram cronometrados cinco minutos e contabilizado o tempo total de permanência do inseto em cada braço do aparelho. Os dados foram analisados pelo teste c², com frequência esperada de 25%. Estudou-se o rendimento de óleo essencial das três espécies de plantas, 30 e 60 dias após a semeadura, utilizando-se do método de hidrodestilação. A composição química dos óleos foi determinada por cromatografia gasosa acoplada a espectrômetro de massas. Verificou-se que adultos virgens têm preferência por plantas de coentro, enquanto os acasalados preferem plantas de erva-doce, ambas coletadas aos 30 dias. Plantas com 60 dias não proporcionaram resposta atrativa aos adultos de C. externa. O rendimento de óleo tendeu a aumentar com o desenvolvimento fenológico da planta. A composição química do óleo de coentro revelou, como componentes majoritários, o (2E)-decenal e decanal e, para erva-doce, a maior concentração foi de (E)-anetol.

Seleção de progênies do cruzamento entre cultivares de cafeeiros Icatu e Catuaí

Os programas de melhoramento devem ser dinâmicos e contínuos, de modo a disponibilizarem sempre novos cultivares de cafeeiros, com fatores de resistência complexos, que se constituam em eficientes barreiras para as novas raças fisiológicas do fungo H. vastatrix. Por essa razão, objetivou-se, com este trabalho, avaliar o comportamento agronômico, a resistência à ferrugem e a ocorrência de cercosporiose em progênies, provenientes do cruzamento dos grupos de cultivares Catuaí e Icatu, em dois ambientes de Minas Gerais. Os experimentos foram implantados nas Fazendas Experimentais da EPAMIG, em São Sebastião do Paraíso e Três Pontas - MG. O delineamento experimental adotado foi o látice quadrado 6 x 6, com 3 repetições. Nos dois ambientes, foram realizadas avaliações de produtividade, vigor vegetativo, percentagem de grãos com peneira 17 acima e reação à ferrugem e à cercosporiose, nos anos agrícolas 2009/2010 e 2010/2011. O desempenho das progênies avaliadas, para produtividade, em Três Pontas, foi superior quando comparado ao de São Sebastião do Paraíso. A progênie H 6-47-10 Cova 3 destacou-se entre as mais promissoras, sugerindo boas possibilidades de progresso genético na seleção.