12 research outputs found

    EB1 Is Required for Spindle Symmetry in Mammalian Mitosis

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    Most information about the roles of the adenomatous polyposis coli protein (APC) and its binding partner EB1 in mitotic cells has come from siRNA studies. These suggest functions in chromosomal segregation and spindle positioning whose loss might contribute to tumourigenesis in cancers initiated by APC mutation. However, siRNA-based approaches have drawbacks associated with the time taken to achieve significant expression knockdown and the pleiotropic effects of EB1 and APC gene knockdown. Here we describe the effects of microinjecting APC- or EB1- specific monoclonal antibodies and a dominant-negative EB1 protein fragment into mammalian mitotic cells. The phenotypes observed were consistent with the roles proposed for EB1 and APC in chromosomal segregation in previous work. However, EB1 antibody injection also revealed two novel mitotic phenotypes, anaphase-specific cortical blebbing and asymmetric spindle pole movement. The daughters of microinjected cells displayed inequalities in microtubule content, with the greatest differences seen in the products of mitoses that showed the severest asymmetry in spindle pole movement. Daughters that inherited the least mobile pole contained the fewest microtubules, consistent with a role for EB1 in processes that promote equality of astral microtubule function at both poles in a spindle. We propose that these novel phenotypes represent APC-independent roles for EB1 in spindle pole function and the regulation of cortical contractility in the later stages of mitosis. Our work confirms that EB1 and APC have important mitotic roles, the loss of which could contribute to CIN in colorectal tumour cells

    The potential of hyperspectral images and partial least square regression for predicting total carbon, total nitrogen and their isotope composition in forest litterfall samples

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    Hosseini Bai, S ORCiD: 0000-0001-8646-6423Purpose: The main objective of this study was to examine the potential of using hyperspectral image analysis for prediction of total carbon (TC), total nitrogen (TN) and their isotope composition (δ13C and δ15N) in forest leaf litterfall samples. Materials and methods: Hyperspectral images were captured from ground litterfall samples of a natural forest in the spectral range of 400–1700 nm. A partial least-square regression model (PLSR) was used to correlate the relative reflectance spectra with TC, TN, δ13C and δ15N in the litterfall samples. The most important wavelengths were selected using β coefficient, and the final models were developed using the most important wavelengths. The models were, then, tested using an external validation set. Results and discussion: The results showed that the data of TC and δ13C could not be fitted to the PLSR model, possibly due to small variations observed in the TC and δ13C data. The model, however, was fitted well to TN and δ15N. The cross-validation R2cv of the models for TN and δ15N were 0.74 and 0.67 with the RMSEcv of 0.53% and 1.07‰, respectively. The external validation R2ex of the prediction was 0.64 and 0.67, and the RMSEex was 0.53% and 1.19 ‰, for TN and δ15N, respectively. The ratio of performance to deviation (RPD) of the predictions was 1.48 and 1.53, respectively, for TN and δ15N, showing that the models were reliable for the prediction of TN and δ15N in new forest leaf litterfall samples. Conclusions: The PLSR model was not successful in predicting TC and δ13C in forest leaf litterfall samples using hyperspectral data. The predictions of TN and δ15N values in the external litterfall samples were reliable, and PLSR can be used for future prediction. © 2017, Springer-Verlag GmbH Germany
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