Inhibition of Aurora kinases enhances chemosensitivity to temozolomide and causes radiosensitization in glioblastoma cells

Glioblastoma remains one of the most devastating human malignancies, and despite therapeutic advances, there are no drugs that significantly improve the patient survival. Altered expression of the Aurora kinases was found in different malignancies, and their inhibition has been studied in cancer therapy. In this study, we analyzed the expression of Aurora A and Aurora B in glioblastoma samples and also analyzed whether the effects of Aurora kinase inhibition were associated with temozolomide or not on cell lines and primary cultures of glioblastoma. RT-PCR assays were used to determine the mRNA expression in glioblastoma tumor samples and in the cell lines. Cell proliferation was measured by XTT assay, and apoptosis was determined by flow cytometry. Drug combination analyses were made based in Chou-Talalay method. Gamma radiation for clonogenic survival used the doses of 2, 4 and 6 Gy. Changes in Aurora B level were assessed by Western blot analysis. Aurora A and B were expressed in glioblastoma samples as well as in the glioblastoma cell lines (n = 6). Moreover, ZM447439, a selective Aurora kinase inhibitor, decreased the proliferation separately and synergistically with temozolomide in primary cultures and cell lines of glioblastoma. ZM also enhanced the effects of radiation on the two cell lines studied (U343 and U251), mainly when associated with TMZ in U343 cells. Treatment with ZM induced apoptotic cell death and diminished Aurora B protein level. These data suggest that Aurora kinase inhibition may be a target for glioblastoma treatment and could be used as adjuvant to chemo- and radiotherapy.

Nd bulk isotopes and 40Ar/39Ar ages of individual minerals from Aurora and Wilkes sub-glacial basin sediments

The Wilkes and Aurora basins are large, low-lying sub-glacial basins that may cause areas of weakness in the overlying East Antarctic ice sheet. Previous work based on ice-rafted debris (IRD) provenance analyses found evidence for massive iceberg discharges from these areas during the late Miocene and Pliocene. Here we characterize the sediments shed from the inferred areas of weakness along this margin (94°E to 165°E) by measuring40Ar/39Ar ages of 292 individual detrital hornblende grains from eight marine sediment core locations off East Antarctica and Nd isotopic compositions of the bulk fine fraction from the same sediments. We further expand the toolbox for Antarctic IRD provenance analyses by exploring the application of 40Ar/39Ar ages of detrital biotites; biotite as an IRD tracer eliminates lithological biases imposed by only analyzing hornblendes and allows for characterization of samples with low IRD concentrations. Our data quadruples the number of detrital 40Ar/39Ar ages from this margin of East Antarctica and leads to the following conclusions: (1) Four main sectors between the Ross Sea and Prydz Bay, separated by ice drainage divides, are distinguishable based upon the combination of 40Ar/39Ar ages of detrital hornblende and biotite grains and the e-Nd of the bulk fine fraction; (2) 40Ar/39Ar biotite ages can be used as a robust provenance tracer for this part of East Antarctica; and (3) sediments shed from the coastal areas of the Aurora and Wilkes sub-glacial basins can be clearly distinguished from one another based upon their isotopic fingerprints.

Androgen-targeted therapy-induced epithelial mesenchymal plasticity and neuroendocrine transdifferentiation in prostate cancer : an opportunity for intervention

Androgens regulate biological pathways to promote proliferation, differentiation, and survival of benign and malignant prostate tissue. Androgen receptor (AR) targeted therapies exploit this dependence and are used in advanced prostate cancer to control disease progression. Contemporary treatment regimens involve sequential use of inhibitors of androgen synthesis or AR function. Although targeting the androgen axis has clear therapeutic benefit, its effectiveness is temporary, as prostate tumor cells adapt to survive and grow. The removal of androgens (androgen deprivation) has been shown to activate both epithelial-to-mesenchymal transition (EMT) and neuroendocrine transdifferentiation (NEtD) programs. EMT has established roles in promoting biological phenotypes associated with tumor progression (migration/invasion, tumor cell survival, cancer stem cell-like properties, resistance to radiation and chemotherapy) in multiple human cancer types. NEtD in prostate cancer is associated with resistance to therapy, visceral metastasis, and aggressive disease. Thus, activation of these programs via inhibition of the androgen axis provides a mechanism by which tumor cells can adapt to promote disease recurrence and progression. Brachyury, Axl, MEK, and Aurora kinase A are molecular drivers of these programs, and inhibitors are currently in clinical trials to determine therapeutic applications. Understanding tumor cell plasticity will be important in further defining the rational use of androgen-targeted therapies clinically and provides an opportunity for intervention to prolong survival of men with metastatic prostate cancer.

Activated oncogenic pathways and therapeutic targets in extranodal nasal-type NK/T cell lymphoma revealed by gene expression profiling

We performed comprehensive genome-wide gene expression profiling (GEP) of extranodal nasal-type natural killer/T-cell lymphoma (NKTL) using formalin-fixed, paraffin-embedded tissue (n = 9) and NK cell lines (n = 5) in comparison with normal NK cells, with the objective of understanding the oncogenic pathways involved in the pathogenesis of NKTL and to identify potential therapeutic targets. Pathway and network analysis of genes differentially expressed between NKTL and normal NK cells revealed significant enrichment for cell cycle-related genes and pathways, such as PLK1, CDK1, and Aurora-A. Furthermore, our results demonstrated a pro-proliferative and anti-apoptotic phenotype in NKTL characterized by activation of Myc and nuclear factor kappa B (NF-kappa B), and deregulation of p53. In corroboration with GEP findings, a significant percentage of NKTLs (n = 33) overexpressed c-Myc (45.4%), p53 (87.9%), and NF-kappa B p50 (67.7%) on immunohistochemistry using a tissue microarray containing 33 NKTL samples. Notably, overexpression of survivin was observed in 97% of cases. Based on our findings, we propose a model of NKTL pathogenesis where deregulation of p53 together with activation of Myc and NF-kappa B, possibly driven by EBV LMP-1, results in the cumulative up-regulation of survivin. Down-regulation of survivin with Terameprocol (EM-1421, a survivin inhibitor) results in reduced cell viability and increased apoptosis in tumour cells, suggesting that targeting survivin may be a potential novel therapeutic strategy in NKTL. Copyright (C) 2011 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

A NOVEL FUNCTION FOR AURORA B KINASE IN THE REGULATION OF P53 BY PHOSPHORYLATION

The mitotic kinase Aurora B plays a pivotal role in mitosis and cytokinesis and governs the spindle assembly checkpoint which ensures correct chromosome segregation and normal progression through mitosis. Aurora B is overexpressed in breast and other cancers and may be an important molecular target for chemotherapy. Tumor suppressor p53 is the guardian of the genome and an important negative regulator of the cell cycle. Previously, it was unknown whether Aurora B and p53 had mutual regulation during the cell cycle. A small molecule specific inhibitor of Aurora B, AZD1152, gave us an indication that Aurora B negatively impacted p53 during interphase and mitosis. Here, we show the antineoplastic activity of AZD1152 in six human breast cancer cell lines, three of which overexpress HER2. AZD1152 specifically inhibited Aurora B kinase activity, thereby causing mitotic catastrophe, polyploidy and apoptosis, which in turn led to apoptotic death. Further, AZD1152 administration efficiently suppressed tumor growth in orthotopic and metastatic breast cancer cell xenograft models. Notably, it was found that the protein level of Aurora B kinase declined after inhibition of Aurora B kinase activity. Investigation of the underlying mechanism suggested that AZD1152 accelerated the protein turnover of Aurora B by enhancing its ubiquitination. As a consequence of inhibition of Aurora B, p53 levels were increased in tissue culture and murine models. This hinted at a possible direct interaction between p53 and Aurora B. Indeed, it was found that p53 and Aurora B exist in complex and interact directly during interphase and at the centromere in mitosis. Further, Aurora B was shown to phosphorylate p53 at several serine/threonine residues in the DNA binding domain and these events caused downregulation of p53 levels via ubiquitination mediated by Mdm2. Importantly, phosphorylation of threonine 211 was shown to reduce p53’s transcriptional activity while other phosphorylation sites did not. On a functional level, Aurora B was shown to reduce p53’s capacity to mediate apoptosis in response to the DNA damaging agent, cisplatin. These results define a novel mechanism for p53 inactivation by Aurora B and imply that oncogenic hyperactivation or overexpression of Aurora B may compromise p53’s tumor suppressor function.

Radioautographic method of determination of number of bacteria in water-bodies [Translation from: Informatsionnyi Byulleten Biologiya Vnutrennikh Vod No.26, 65-67, 1975]

The radioautographic method of determination of the number of autotrophic microorganisms was initially suggested for counting methane-oxidizing bacteria. With the help of this method colonies of hydrogen-oxidizing bacteria are differentiated even more clearly from heterotrophic. Under laboratory conditions it was shown that colonies grown on membrane filters from a pure culture of thionic bacteria on a nutrient medium with radio- active carbonate, give better prints on film. This method was tested by the authors for determining the number of these bacteria in the meromictic Lake Vae de San Juan during the expedition to Cuba in the summer of 1973. The study showed that that the thionic bacteria are found throughout the pelagial. It proved that the thionic bacteria can be well considered in water-bodies by the radioautographic method.

miR-433 overexpression attenuates the spindle assembly checkpoint response to paclitaxel

Paclitaxel is a microtubule inhibitory chemotherapeutic drug that is increasingly used for the treatment of solid tumours. In vitro studies have demonstrated that attenuating the spindle assemble checkpoint (SAC) alters the post-mitotic responses to paclitaxel. Furthermore, the aberrant expression of a number of the SAC proteins, MAD2, BUBR1, and Aurora A kinase, are associated with poor patient prognosis. We have identified a microRNA, miR-433, that regulates the expression of MAD2. Overexpression of miR-433 in Hela cells induced downregulation of MAD2 mRNA and protein expression. We have also shown that Hela cells overexpressing miR-433 and treated with paclitaxel are no longer capable of cyclin B stabilisation, and thus have lost the ability to activate the SAC in response to paclitaxel. In addition, cell viability assays showed that Hela cells overexpressing miR-433 and treated with paclitaxel have an attenuated response to paclitaxel compared with microRNA scrambled controls. We have characterised the levels of miR-433, MAD2 gene expression and MAD2 protein levels in a cohort of ovarian cancer cell lines. Cell viability assays on this cohort revealed that responsiveness to paclitaxel is associated with high MAD2 protein expression and lower miR-433 expression. We hypothesise that the expression of miR-433 when deregulated in cancer leads to altered MAD2 expression and a compromised SAC, a key feature underlying drug resistance to paclitaxel. In a pilot study of paired human breast tumour and normal breast tissue samples we have shown that expression levels of miR-433 are elevated in cancer tissue. Targeting this microRNA in cancer may improve the efficacy of paclitaxel in treating breast cancer and ovarian cancer.

Reação de clones de umezeiro (Prunus mume Sieb. et Zucc.) e cultivares de pessegueiro [Prunus persica (L.) Batsch] ao nematóide anelado Mesocriconema xenoplax (Nemata: Criconematidae)

Com o objetivo de avaliar a reação de clones de umezeiro (Prunus mume Sieb. et Zucc.) e cultivares de pessegueiro [Prunus persica (L.) Batsch] ao nematóide anelado Mesocriconema xenoplax (Raski) Loof & de Grise, realizou-se o presente estudo em casa de vegetação do Departamento de Fitossanidade da FCAV/UNESP, Câmpus de Jaboticabal-SP. As plantas foram mantidas em vasos de cerâmica com 6 litros de capacidade, contendo uma mistura de solo e areia (1:1, v/v), previamente autoclavada a 121°C e 1kgf.cm-2 por 2 horas. Cada planta foi inoculada com 10mL de uma suspensão de 200 M. xenoplax por mL. Com os resultados obtidos, após 105 dias da inoculação, pode-se concluir que os Clones 05; 10 e 15 de umezeiro e as cultivares Okinawa e Aurora-1 de pessegueiro são suscetíveis a M. xenoplax. A cultivar Aurora-1 apresentou maior Fator de Reprodução (93,06).

Relação entre as características químicas e o valor dos pêssegos comercializados pelo sistema veiling frutas Holambra em Paranapanema-SP

O valor de comercialização do pêssego é reflexo da demanda e de sua apreciação pelo consumidor. A compreensão da diferença de valor entre os frutos das diferentes cultivares e da sua relação com as características que determinam o gosto do fruto, torna possível o estabelecimento de uma estratégia de comercialização visando ao aumento no consumo e na receita do produtor, além de dar subsídios aos programas de melhoramento genético. Neste trabalho, foram avaliadas as características de gosto de duas cultivares, 'Douradão' e 'Tropic Beauty', e de duas séries de cultivares, 'Aurora' e 'Dourado', que são as cultivares de pêssego mais produzidas no município de Paranapanema, maior produtor do Estado de São Paulo. Trabalhou-se com os valores de comercialização do leilão reverso, ou veiling, da Cooperativa Agroindustrial Holambra, no período de maior oferta do produto, entre 15 de outubro a 15 de novembro de 2004. A caracterização do gosto dos frutos foi feita através da determinação dos conteúdos de sólidos solúveis (SS) e de acidez titulável (AT), e da relação SS/AT dos frutos comercializados no Entreposto Terminal de São Paulo da CEAGESP. As médias dos conteúdos de sólidos solúveis (SS) não se mostraram significativamente diferentes. A acidez titulável (AT) e a relação SS/AT apresentaram valores médios significativamente diferentes. A cultivar 'Douradão' apresentou a maior relação SS/AT, seguida das séries varietais 'Dourado' e 'Aurora' e da cultivar Tropic Beauty. As diferenças na relação SS/AT não determinaram diferenças significativas no valor dos frutos do mesmo calibre, nos quatro materiais estudados. O valor de comercialização dos produtos mostrou-se significativamente afetado pelos calibres.

Produção e sazonalidade de pessegueiro e nectarineira sob florescimento espontâneo e com cianamida hidrogenada e óleo mineral

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Eficácia do carfentrazone-ethyl no controle de plantas aquáticas latifoliadas em caixas-d'água

Atualmente, alguns herbicidas estão sendo desenvolvidos para o controle de plantas daninhas aquáticas. O objetivo deste trabalho foi avaliar a eficácia do carfentrazone-ethyl em ambiente aquático para o controle pós-emergente de aguapé, alface-d'água e salvínia. O trabalho foi desenvolvido em caixas-d'água, no período de julho a setembro de 2004, no NUPAM - FCA/UNESP, em Botucatu. O delineamento experimental adotado foi o de blocos ao acaso, com sete tratamentos e quatro repetições, sendo as unidades experimentais constituídas pelas caixasd'água. Os tratamentos foram os seguintes: testemunha sem herbicida; Aurora 400 CE (75, 150 e 300 mL ha-1); Roundup (3,0 L ha-1), Aurora 400 CE + Roundup (75 mL + 3,0 L ha-1) e Aurora 400 CE + Arsenal N.A. (75 mL + 2,0 L ha-1). Observou-se que o tratamento Aurora 400 CE (300 mL ha-1) é altamente eficaz no controle de alface-d'água (Pistia stratiotes); o tratamento Roundup (3,0 L ha-1) é altamente eficaz no controle de aguapé (Eichhornia crassipes); o tratamento Aurora 400 CE + Roundup (75 mL + 3,0 L ha-1) é eficaz no controle de aguapé (E. crassipes), alface-d'água (P. stratiotes) e salvínia (Salvinia auriculata); e o tratamento Aurora 400 CE + Arsenal (75 mL + 2,0 L ha-1) é eficaz no controle de aguapé (E. crassipes) e alface-d'água (P. stratiotes). A mistura Aurora 400 CE + Roundup (75 mL + 3,0 L ha-1) apresentou-se viável e foi o único tratamento eficaz no controle das três espécies estudadas.

Composição acarina em diferentes cultivares de pessegueiros [Prunus persica (L.)], em Presidente Prudente, estado de São Paulo

Objetivou-se, nesta pesquisa, estudar a ocorrência natural de ácaros fitófagos e predadores em diferentes cultivares de pessegueiro, no município de Presidente Prudente-SP, Brasil. O estudo foi realizado no período de dezembro de 2002 a fevereiro de 2006. Amostras quinzenais de 72 folhas foram coletadas ao acaso, de pessegueiros das cultivares Talismã, Doçura 2, Dourado 2, Tropical, Aurora 1 e Aurora 2. Coletou-se um total de 2.594 ácaros, sendo 2.092 fitófagos, 403 predadores e 99 de hábitos alimentares pouco conhecidos, com 35 espécies de ácaros de 16 famílias. Aculus fockeui ocorreu de maneira esporádica, não causando danos visíveis às plantas. A família Phytoseiidae apresentou a maior abundância e o maior número de indivíduos. O predador Euseius citrifolius foi o mais abundante. Não houve preferência dos ácaros nas cultivares de pessegueiro avaliadas.

Frutificação, características físicas de frutos e produtividade em cultivares e seleções de pessegueiro em Vista Alegre do Alto-SP

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Infestación natural de Lonchaeidae (Diptera) en variedades de melocotón

Se estudió la ocurrencia de Lonchaeidae en variedades de melocotón, conducidos sobre los patrones ‘Okinawa’ y Umê: Tropical, Ouromel 3, Jóia 4, Régis, Talismã, Aurora 2, Aurora 1, Dourado 2 y Doçura 2. Se colectaron muestras de 30 frutos por planta en el Banco de Germoplasma en Presidente Prudente, Brasil. Se obtuvieron 633 especímenes de Lonchaeidae; 394 de ellos en frutos de variedades sobre ‘Okinawa’ y 239 sobre Umê. Se capturaron especies como Neosilba zadolicha, N. inesperata, N. pendula, N. certa y Neosilba spp. (hembras). N. zadolicha y N. inesperata, se observaron en 77,78% de las muestras de las variedades sobre ‘Okinawa’. Sobre el patrón Umê la mayor incidencia fue de N. zadolicha , N. inesperata y N. pendula (55,6%, 33,3% y 33,3% respectivamente). Las variedades Ouromel 3, Talismã, Doçura 2 y Aurora 2 presentaron mayores infestaciones por Lonchaeidae. N. certa tuvo una menor incidencia y sólo se observó en la variedad Doçura 2 sobre ‘Okinawa’. Plantas con mayor número de frutos presentaron mayor incidencia de moscas por fruto; sin embargo, no hubo correlación entre peso del fruto y número de moscas. No se observó diferencia para peso del fruto y número de moscas por fruto entre los dos patrones, ‘Okinawa’ y Umê. Melocotones crecidos en los patrones ‘Okinawa’ y Umê en Presidente Prudente están infestados por especies de Lonchaeidae que presentan preferencia independientemente del peso de los frutos. El aporte de este trabajo al conocimiento de la comunidad de especies de Lonchaeidae ofrece una base para su control.