Temporally distinct roles of ATM and ROS in genotoxic- stress-dependent induction and maintenance of cellular senescence

Cells exposed to genotoxic stress induce cellular senescence through a DNA damage response (DDR) pathway regulated by ATM kinase and reactive oxygen species (ROS). Here, we show that the regulatory roles for ATM kinase and ROS differ during induction and maintenance of cellular senescence. Cells treated with different genotoxic agents were analyzed using specific pathway markers and inhibitors to determine that ATM kinase activation is directly proportional to the dose of the genotoxic stress and that senescence initiation is not dependent on ROS or the p53 status of cells. Cells in which ROS was quenched still activated ATM and initiated the DDR when insulted, and progressed normally to senescence. By contrast, maintenance of a viable senescent state required the presence of ROS as well as activated ATM. Inhibition or removal of either of the components caused cell death in senescent cells, through a deregulated ATM-ROS axis. Overall, our work demonstrates existence of an intricate temporal hierarchy between genotoxic stress, DDR and ROS in cellular senescence. Our model reports the existence of different stages of cellular senescence with distinct regulatory networks.

Analysing the contribution of ATM/ATR pathway activation in establishing the premature senescence of E2F1/E2F2-/- bone-marrow-derived macrophages

E2F1 and E2F2 transcription factors have an important role during the regulation of cell cycle. In experiments done with E2F1/E2F2 knockout mice, it has been described that bone-marrow-derived macrophages (BMDM) undergo an early rapid proliferation event related to DNA hyper-replication. As a consequence, DNA damage response (DDR) pathway is triggered and E2F1/E2F2 knockout macrophages enter premature senescence related to G2/M phase arrest. The exact mechanism trough which DNA hyper-replication leads to DDR in absence of E2F1 and E2F2 remains undiscovered. To determine whether the ATR/ATM pathway, the master regulator of G2/M checkpoint, might be the surveillance mechanism in order to regulate uncontrolled proliferation in the DKO model, we monitored and analysis biochemical properties of BMDM cultures in the presence of caffeine, a potent inhibitor of ATM/ATR activity. Our results show that the addition of caffeine abolishes premature senescence in DKO BMDM, stimulates γ-H2AX accumulation and decreases Mcm2 expression.

Avaliaçāo preliminar dos recursos de camarāo de profundidade em Moçambique

Although it is known enough of the biology of shallow+ waters shrimps, knowledge of the deep-waters shrimp is still limited. This is due to the fact that: these resources are relatively new and the study of deep waters species is more difficult and expensive. Some biological informations on the main species of shrimp in depth of Mozambique have been collected during investigation cruises (Sports et al., Pacule, 1987) and through sampling carried out on board commercial vessels (DDR fleet) between 1983 and 1986.

藏酋猴(Macaca thibetana)电刺激采精、精液冻存和精子活力的检测

本文研究中国特有的藏酋猴(Macaca thibetana)电刺激采精、精液冻存及精子活力检测。用电刺激方案[(DDR)_(17-c)V]：延搁(delay)17毫秒、间期(duration)17毫秒、刺激频率17脉冲／秒、单方波变电压连续刺激，对笼养藏酋猴进行电刺激采精，测定精液的各种特征参数，包括每次射精量、液化率、精子密度、运动精子的百分率、死活精子以及异常精子比例。并与其它非人灵长类进行了比较。在精液冻存实验中，通过对不同的防冻液、不同的降温程序及防冻液分散系的比较研究，从而确定了适于藏酋猴精液冻存的冻存方案：PSF-4%的甘油-TH-7.5%的小牛血清(PSF-TH/FBS-G)和PSF-MDM。用前一方案冻存藏酋猴精液，复苏运动度为63.58±0.06%；精子的存活率为90.14±0.03%(n=5)。冻存精液用金黄地鼠裸穿透分析法检测，结果表明具有相应新鲜精液穿透力的51.90±0.08%。后一冻存方案能保存88.00±1.03%的运动精子，但运动寿命较短。研究结果表明：1).刺激方案(DDR)_(17-c)V适于藏酋猴的电刺激采精。2).藏酋猴的精液量和精子数在已有过研究的非人灵长类动物中是最多的，藏酋猴有可能成为较理想的精子生物学研究用的非人灵长类动物。3).脂蛋白对维持精子膜的完整可能有重要作用。4).接近生理条件的防冻剂分散系有助于精液的冻存。5).两种冻存方案PSF-TH/FBS-G和PSF-MDM均适于藏酋猴的精液冻存。

Selénio e a importância para o organismo humano

O selénio (Se) é um micronutriente essencial para o crescimento, desenvolvimento e normal metabolismo dos animais, incluindo o ser humano. É parte integrante de um conjunto de proteínas, as selenoproteínas, com ação antioxidante (protegendo as membranas celulares contra danos dos radicais livres), envolvidas no metabolismo das hormonas da tiróide, na regulação do crescimento e viabilidade celular, nas funções do sistema imune e na reprodução. É introduzido na dieta alimentar (principalmente nas formas de selenometionina e selenocisteína) através das plantas, e de produtos que delas derivam, que assimilam os compostos de selénio presentes no solo. Uma vez que a quantidade de selénio existente nos solos é muito variável, o teor nos alimentos vai depender da sua origem geográfica e, por consequência, a ingestão de selénio varia entre regiões e países. Baixos níveis de selénio estão associados a um declínio na função imune e problemas cognitivos. A deficiência de Se pode também ocasionar problemas musculares e cardiomiopatia. Concentrações reduzidas foram observadas em indíviduos com crises epiléticas e também em casos de pré-eclampsia. A deficiência de selénio pode também desenvolver-se durante a nutrição parenteral. Atualmente, a Dose Diária Recomendada (DDR) é de 55 μg/dia para homens e mulheres adultos e saudáveis. No entanto, existem evidências clínicas de que a ingestão em doses superiores (200-300 μg/dia) pode ter um papel benéfico na prevenção de alguns tipos de cancro e doenças cardiovasculares, na melhoria da resposta imunológica, como neuroprotetor e na fertilidade. O Se desempenha um papel importante na fertilidade masculina, sendo necessário na biossíntese da testosterona e na formação e normal desenvolvimento dos espermatozóides. Em mulheres grávidas o Se, ajuda a prevenir complicações antes e durante o parto e promove o normal desenvolvimento do feto. Como antioxidante o selénio vai combater os danos provocados pelos radicais livres, impedindo que estes exerçam o seu papel prejudicial no organismo. Sendo o sistema imunológico muito suscetível aos danos provocados pelo stress oxidativo, o Se vai exercer efeitos benéficos combatendo os danos por ele causados. Relativamente à capacidade viral, não é possível saber com exatidão qual a quantidade de Se necessária ou concentração ideal no plasma para evitar a ocorrência e desenvolvimento de infeções virais. No entanto, sabe-se que tem um efeito benéfico em pacientes HIV positivos e em indivíduos infetados com o vírus da hepatite (B ou C) contra a progressão para o neoplasia de fígado. Em teoria, a nível cardiovascular, este elemento pode exercer um efeito protetor, embora alguns estudos epidemiológicos não tenham mostrado uma associação clara entre o risco cardiovascular e os níveis selénio. A nível cerebral o Se vai atuar como neuroprotetor, prevenindo o aparecimento de patologias como demência e doença de Alzheimer. Apesar destes indicadores, a maioria dos países europeus, incluindo Portugal, regista uma deficiente ingestão de selénio por parte da população. A suplementação poderá constituir uma opção para garantir os níveis nutricionais recomendados e/ou ser utilizada com o objetivo de prevenir algumas doenças e o envelhecimento. No entanto o selénio pode também ser tóxico se ingerido em excesso, estando a dose máxima admissível fixada em 400 μg/dia. A intoxicação por selénio é chamada selenose e os sintomas comuns incluem: hálito a alho, distúrbios gastrointestinais, perda de cabelo, descamação das unhas, danos neurológicos e fadiga. Assim, atualmente acredita-se que enquanto indivíduos com baixo nível de Se podem obter benefícios da suplementação, esta pode ser prejudicial aqueles com valores normais ou elevados.

The Liedertheater of Wenzel and Mensching: Subversive Use of the Agitprop and Clowning Tradition in the GDR

The East German poet-clowns Hans-Eckardt Wenzel and Steffen Mensching rose to prominence during the GDR's (German Democratic Republic; Deutsche Demokratische Republik, DDR) Peaceful Revolution of autumn 1989 with their cabaret production Letztes aus der Da Da eR. A film adaptation of the production was made by Jörg Foth in 1990, which was finally released on DVD with English subtitles in the United Kingdom and North America in 2009 (Latest from the Da-Da-R). In light of this long-overdue interest in Wenzel & Mensching, this article will attempt to put the work of the duo in historical and aesthetic context. Their use of character, masks, music, and philosophy combined to create the distinctly grotesque world that constituted their Liedertheater performances.

Valved Dataflow For FPGA Memory Hierarchy Synthesis

For modern FPGA, implementation of memory intensive processing applications such as high end image and video processing systems necessitates manual design of complex multilevel memory hierarchies incorporating off-chip DDR and onchip BRAM and LUT RAM. In fact, automated synthesis of multi-level memory hierarchies is an open problem facing high level synthesis technologies for FPGA devices. In this paper we describe the first automated solution to this problem.
By exploiting a novel dataflow application modelling dialect, known as Valved Dataflow, we show for the first time how, not only can such architectures be automatically derived, but also that the resulting implementations support real-time processing for current image processing application standards such as H.264. We demonstrate the viability of this approach by reporting the performance and cost of hierarchies automatically generated for Motion Estimation, Matrix Multiplication and Sobel Edge Detection applications on Virtex-5 FPGA.

A theoretical examination of known and hypothetical clathrate hydrate materials

The recent synthesis of a new hydrogen binary hydrate with the sH structure has highlighted the potential storage capabilities of water clathrates [T. A. Strobel, C. A. Koh, and E. D. Sloan, J. Phys. Chem. B 112, 1885 (2008) and A. R. C. Duarte, A. Shariati, L. J. Rovetto, and C. J. Peters, J. Phys. Chem. B 112, 1888 (2008)]. In this work, the absorption of hydrogen and the promoters used in the experimental work are considered using a simplified model for the host-guest interaction, which allows one to understand the stabilizing effects of multiple help molecules. Two further hypothetical clathrates, which are isostructural with known zeolite structures, are also investigated. It is shown that the energy gained by absorbing adamantane into these two frameworks is far greater than that gained upon absorption of adamantane into the sH structure. Hence, a clathrate with the same topology as the DDR (Sigma 1) zeolite may be synthesizable with adamantane and hydrogen as guest molecules as, in the conditions explored here, this phase appears to be more stable than the sH structure. (C) 2009 American Institute of Physics. [DOI: 10.1063/1.3142503]

Identification and Validation of an Anthracycline/Cyclophosphamide–Based Chemotherapy Response Assay in Breast Cancer

Background: There is no method routinely used to predict response to anthracycline and cyclophosphamide–based chemotherapy in the clinic; therefore patients often receive treatment for breast cancer with no benefit. Loss of the Fanconi anemia/BRCA (FA/BRCA) DNA damage response (DDR) pathway occurs in approximately 25% of breast cancer patients through several mechanisms and results in sensitization to DNA-damaging agents. The aim of this study was to develop an assay to detect DDR-deficient tumors associated with loss of the FA/BRCA pathway, for the purpose of treatment selection.

Methods: DNA microarray data from 21 FA patients and 11 control subjects were analyzed to identify genetic processes associated with a deficiency in DDR. Unsupervised hierarchical clustering was then performed using 60 BRCA1/2 mutant and 47 sporadic tumor samples, and a molecular subgroup was identified that was defined by the molecular processes represented within FA patients. A 44-gene microarray-based assay (the DDR deficiency assay) was developed to prospectively identify this subgroup from formalin-fixed, paraffin-embedded samples. All statistical tests were two-sided.

Results: In a publicly available independent cohort of 203 patients, the assay predicted complete pathologic response vs residual disease after neoadjuvant DNA-damaging chemotherapy (5-fluorouracil, anthracycline, and cyclophosphamide) with an odds ratio of 3.96 (95% confidence interval [Cl] =1.67 to 9.41; P = .002). In a new independent cohort of 191 breast cancer patients treated with adjuvant 5-fluorouracil, epirubicin, and cyclophosphamide, a positive assay result predicted 5-year relapse-free survival with a hazard ratio of 0.37 (95% Cl = 0.15 to 0.88; P = .03) compared with the assay negative population.

Conclusions: A formalin-fixed, paraffin-embedded tissue-based assay has been developed and independently validated as a predictor of response and prognosis after anthracycline/cyclophosphamide–based chemotherapy in the neoadjuvant and adjuvant settings. These findings warrant further validation in a prospective clinical study.

Financial reintegration assistance for veterans of the Irish revolution (1916-23): post-conflict policy in an historical setting

In the aftermath of the Irish revolution and Civil War the governments of independent Ireland introduced various compensation schemes to provide financial reintegration assistance to revolutionary veterans. This would be recognised today as part of a programme for DDR. This paper will examine various service and disability pensions paid to veterans in the context of literature on post-conflict reintegration. It will examine various challenges to reintegration in an effort to analyse the success of revolutionary compensation as a post-conflict reintegration mechanism in independent Ireland after 1922.

Mechanistic Rationale to Target PTEN-Deficient Tumor Cells with Inhibitors of the DNA Damage Response Kinase ATM

Ataxia telangiectasia mutated (ATM) is an important signaling molecule in the DNA damage response (DDR). ATM loss of function can produce a synthetic lethal phenotype in combination with tumor-associated mutations in FA/BRCA pathway components. In this study, we took an siRNA screening strategy to identify other tumor suppressors that, when inhibited, similarly sensitized cells to ATM inhibition. In this manner, we determined that PTEN and ATM were synthetically lethal when jointly inhibited. PTEN-deficient cells exhibited elevated levels of reactive oxygen species, increased endogenous DNA damage, and constitutive ATM activation. ATM inhibition caused catastrophic DNA damage, mitotic cell cycle arrest, and apoptosis specifically in PTEN-deficient cells in comparison with wild-type cells. Antioxidants abrogated the increase in DNA damage and ATM activation in PTEN-deficient cells, suggesting a requirement for oxidative DNA damage in the mechanism of cell death. Lastly, the ATM inhibitor KU-60019 was specifically toxic to PTEN mutant cancer cells in tumor xenografts and reversible by reintroduction of wild-type PTEN. Together, our results offer a mechanistic rationale for clinical evaluation of ATM inhibitors in PTEN-deficient tumors.

The Nuclear Oncogene SET Controls DNA Repair by KAP1 and HP1 Retention to Chromatin

Cells experience damage from exogenous and endogenous sources that endanger genome stability. Several cellular pathways have evolved to detect DNA damage and mediate its repair. Although many proteins have been implicated in these processes, only recent studies have revealed how they operate in the context of high-ordered chromatin structure. Here, we identify the nuclear oncogene SET (I2PP2A) as a modulator of DNA damage response (DDR) and repair in chromatin surrounding double-strand breaks (DSBs). We demonstrate that depletion of SET increases DDR and survival in the presence of radiomimetic drugs, while overexpression of SET impairs DDR and homologous recombination (HR)-mediated DNA repair. SET interacts with the Kruppel-associated box (KRAB)-associated co-repressor KAP1, and its overexpression results in the sustained retention of KAP1 and Heterochromatin protein 1 (HP1) on chromatin. Our results are consistent with a model in which SET-mediated chromatin compaction triggers an inhibition of DNA end resection and HR.

AURKA overexpression accompanies dysregulation of DNA-damage response genes in invasive urothelial cell carcinoma

Invasive urothelial cell carcinoma (UCC) is characterized by increased chromosomal instability and follows an aggressive clinical course in contrast to non-invasive disease. To identify molecular processes that confer and maintain an aggressive malignant phenotype, we used a high-throughput genome-wide approach to interrogate a cohort of high and low clinical risk UCC tumors. Differential expression analyses highlighted cohesive dysregulation of critical genes involved in the G(2)/M checkpoint in aggressive UCC. Hierarchical clustering based on DNA Damage Response (DDR) genes separated tumors according to a pre-defined clinical risk phenotype. Using array-comparative genomic hybridization, we confirmed that the DDR was disrupted in tumors displaying high genomic instability. We identified DNA copy number gains at 20q13.2-q13.3 (AURKA locus) and determined that overexpression of AURKA accompanied dysregulation of DDR genes in high risk tumors. We postulated that DDR-deficient UCC tumors are advantaged by a selective pressure for AURKA associated override of M phase barriers and confirmed this in an independent tissue microarray series. This mechanism that enables cancer cells to maintain an aggressive phenotype forms a rationale for targeting AURKA as a therapeutic strategy in advanced stage UCC.

NanoStreams: Codesigned Microservers for Edge Analytics in Real Time

NanoStreams explores the design, implementation,and system software stack of micro-servers aimed at processingdata in-situ and in real time. These micro-servers can serve theemerging Edge computing ecosystem, namely the provisioningof advanced computational, storage, and networking capabilitynear data sources to achieve both low latency event processingand high throughput analytical processing, before consideringoff-loading some of this processing to high-capacity datacentres.NanoStreams explores a scale-out micro-server architecture thatcan achieve equivalent QoS to that of conventional rack-mountedservers for high-capacity datacentres, but with dramaticallyreduced form factors and power consumption. To this end,NanoStreams introduces novel solutions in programmable & con-figurable hardware accelerators, as well as the system softwarestack used to access, share, and program those accelerators.Our NanoStreams micro-server prototype has demonstrated 5.5×higher energy-efficiency than a standard Xeon Server. Simulationsof the microserver’s memory system extended to leveragehybrid DDR/NVM main memory indicated 5× higher energyefficiencythan a conventional DDR-based system. 

Nachrichtendienstpsychologie 3

Der Versuch, nachrichtendienstliche Fragestellungen unter psychologischen Aspekten zu betrachten, ist auf ein positives Interesse gestoßen. In den beiden Bänden Nachrichtendienstpsychologie 1 (Litzcke, 2003) und 2 (Schwan, 2004) der FH Bund erschienen eine Reihe von Aufsätzen, die ein breites Themenspektrum behandeln. Mit Band 1 wurde eine Struktur eingeführt, die sich bewährt hat, und die daher auch für Band 3 beibehalten wird. Die Beiträge des Bandes 3 lassen sich wie folgt zuordnen: 1) Anwendungen vorhandener Konzepte der Psychologie und aus Nachbardisziplinen in den Nachrichtendiensten. Siehe hierzu die Beiträge von Scherer zur Moderation von Qualitätszirkeln im Bundesnachrichtendienst, von Damm und Litzcke zur Akzeptanz des Sicherheitsakteurs „Europa“, von Schwan zur Bedeutung aggressiven Verhaltens für die Arbeit der Nachrichtendienste, von Löhr zum kognitiven Interview und von Wiesen zu den Möglichkeiten einer Gesprächssteuerung. 2) Übertragung psychologischer Methoden auf das Forschungsgebiet Nachrichtendienste zur Gewinnung neuer Ergebnisse / Konzepte. Siehe hierzu die Beiträge von Müller-Enbergs zu den Motiven für eine geheimdienstliche oder nachrichtendienstliche Kooperation bei den inoffiziellen Mitarbeitern der DDR-Staatssicherheit, von Horn zu den Einsatzmöglichkeiten der Operativen Fallanalyse und von Schmalzl zum Konstrukt Einsatzkompetenz. 3) Generierung eigenständiger psychologischer Erkenntnisse und Methoden im Bereich Nachrichtendienste. Dieser Rubrik ist kein Beitrag des Bandes 3 zuzuordnen. Der Abschlussbeitrag von Freitag zur Anwendbarkeit von Täuschungsprinzipien der Zauberkunst in der nachrichtendienstlichen Arbeit entzieht sich dieser Struktur. Der Beitrag ist mit Augenzwinkern geschrieben und soll auch so gelesen werden. Die Themen der Aufsätze sind nicht auf rein nachrichtendienstliche Aufgabenstellungen beschränkt. So werden beispielsweise auch Querschnittsproblematiken behandelt, die nicht nur einem Arbeitsbereich innerhalb der Nachrichtendienste zuzuordnen wären. Damit werden auch Themen behandelt, die ebenso in anderen Behörden und Institutionen von praktischer Relevanz sein können. So ist beispielsweise die Einrichtung von Qualitätszirkeln kein spezifisches Thema der Nachrichtendienste. Da sich aber auch Nachrichtendienste mit der Qualitätssteigerung ihrer Arbeit befassen, ist es angezeigt, entsprechende Ausarbeitungen in die Reihe Nachrichtendienstpsychologie zu integrieren. Auch in Band 3 werden interdisziplinäre Ansätze und Verknüpfungen mit der Polizeipsychologie vorgestellt, auch wenn die dort entwickelten Erkenntnisse und Methoden nicht ungeprüft auf nachrichtendienstliche Aufgabenstellungen übertragen werden können. Gleichwohl können sich aus dem Kontakt zur Polizeipsychologie Anregungen für die Nachrichtendienstpsychologie ergeben.