An essential function for the ATR-Activation-Domain (AAD) of TopBP1 in mouse development and cellular senescence

ATR activation is dependent on temporal and spatial interactions with partner proteins. In the budding yeast model, three proteins – Dpb11TopBP1, Ddc1Rad9 and Dna2 - all interact with and activate Mec1ATR. Each contains an ATR activation domain (ADD) that interacts directly with the Mec1ATR:Ddc2ATRIP complex. Any of the Dpb11TopBP1, Ddc1Rad9 or Dna2 ADDs is sufficient to activate Mec1ATR in vitro. All three can also independently activate Mec1ATR in vivo: the checkpoint is lost only when all three AADs are absent. In metazoans, only TopBP1 has been identified as a direct ATR activator. Depletion-replacement approaches suggest the TopBP1-AAD is both sufficient and necessary for ATR activation. The physiological function of the TopBP1 AAD is, however, unknown. We created a knock-in point mutation (W1147R) that ablates mouse TopBP1-AAD function. TopBP1-W1147R is early embryonic lethal. To analyse TopBP1-W1147R cellular function in vivo, we silenced the wild type TopBP1 allele in heterozygous MEFs. AAD inactivation impaired cell proliferation, promoted premature senescence and compromised Chk1 signalling following UV irradiation. We also show enforced TopBP1 dimerization promotes ATR-dependent Chk1 phosphorylation. Our data suggest that, unlike the yeast models, the TopBP1-AAD is the major activator of ATR, sustaining cell proliferation and embryonic development

Central nervous system and muscular bundles preserved in a 240 million year old giant bristletail (Archaeognatha: Machilidae)

Among the incomparably diverse group of insects no cases of central nervous system (CNS) preservation have been so far described in compression fossils. A third of the fossil insects collected from a 240-239 million year old (Ma) level at Monte San Giorgio UNESCO World Heritage (SwitzerlandItaly) underwent phosphatization, resulting in the extraordinary preservation of soft tissues. Here we describe Gigamachilis triassicus gen. et sp. nov. (Archaeognatha: Machiloidea: Machilidae) that, with an estimated total length of similar to 80 millimeters, represents the largest apterygote insect ever recorded. The holotype preserves: (i) components of the CNS represented by four abdominal ganglia, optic lobes with neuropils and compound retina;(ii) muscular bundles. Moreover, G. triassicus, possessing morphological features that prompt its assignment to the extant archaeognathan ingroup Machilidae, places the origin of modern lineages to Middle Triassic. Interestingly, at Monte San Giorgio, in the same stratigraphic unit the modern morphology of G. triassicus co-occurs with the ancient one represented by Dasyleptus triassicus (Archaeognatha: dagger Monura). Comparing these two types of body organization we provide a new reconstruction of the possible character evolution leading towards modern archaeognathan forms, suggesting the acquisition of novel features in a lineage of apterygote insects during the Permian or the Lower Triassic

Multispectral optoacoustic tomography (MSOT) for imaging the particle size-dependent intratumoral distribution of polymeric micelles

Zhaoqing Cong, Feifei Yang, Li Cao, Han Wen, Taotao Fu, Siqi Ma, Chunyu Liu, Lihui Quan, Yonghong Liao Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Haidian District, Beijing 100193, People’s Republic of China Purpose: This study proposes the utilization of multispectral optoacoustic tomography (MSOT) to investigate the intratumoral distribution of polymeric micelles and effect of size on the biodistribution and antitumor efficacy (ATE). Materials and methods: Docetaxel and/or optoacoustic agent-loaded polymeric micelles (with diameters of 22, 48, and 124 nm) were prepared using amphiphilic block copolymers poly (ethylene glycol) methyl ether-block-poly (D,L lactide) (PEG2000–PDLLAx). Subcutaneous 4T1 tumor-bearing mice were monitored with MSOT imaging and IVIS® Spectrum in vivo live imaging after tail vein injection of micelles. The in vivo results and ex vivo confocal imaging results were then compared. Next, ATE of the three micelles was found and compared. Results: We found that MSOT imaging offers spatiotemporal and quantitative information on intratumoral distribution of micelles in living animals. All the polymeric micelles rapidly extravasated into tumor site after intravenous injection, but only the 22-nm micelle preferred to distribute into the inner tumor tissues, leading to a superior ATE than that of 48- and 124-nm micelles. Conclusion: This study demonstrated that MSOT is theranostically a powerful imaging modality, offering quantitative information on size-dependent spatiotemporal distribution patterns after the extravasation of nanomedicine from tumor blood vessels. Keywords: multispectral optoacoustic tomography, MSOT, polymeric micelle, in vivo imaging, intratumoral distribution, particle size, tumor mode

Visualization of arrangements of carbon atoms in graphene layers by Raman mapping and atomic-resolution TEM

10.1038/srep01195Scientific Reports3