An intriguing set of points of a generalised quadrangle was introduced in [J. Bamberg, M. Law, T. Penttila, Tight sets and m-ovoids of generalised quadrangles, Combinatorica, in press] as a unification of the pre-existing notions of tight set and m-ovoid. It was shown in [J. Bamberg, M. Law, T. Penttila, Tight sets and m-ovoids of generalised quadrangles, Combinatorica, in press] that every intriguing set of points in a finite generalised quadrangle is a tight set or an m-ovoid (for some m). Moreover, it was shown that an m-ovoid and an i-tight set of a common generalised quadrangle intersect in mi points. These results yielded new proofs of old results, and in this paper, we study the natural analogue of intriguing sets in finite polar spaces of higher rank. In particular, we use the techniques developed in this paper to give an alternative proof of a result of Thas [J.A. Thas, Ovoids and spreads of finite classical polar spaces, Geom. Dedicata 10 (1-4) (1981) 135-143] that there are no ovoids of H(2r, q 2 ), Q − (2r + 1, q), and W(2r − 1, q) for r > 2. We also strengthen a result of Drudge on the non-existence of tight sets in W(2r − 1, q), H(2r + 1, q 2 ), and Q + (2r + 1, q), and we give a new proof of a result of De Winter, Luyckx, and Thas [S. De Winter, J.A. Thas, SPG-reguli satisfying the polar property and a new semipartial geometry, Des. Codes Cryptogr. 32 (1-3) (2004) 153-166; D. Luyckx, m-Systems of finite classical polar spaces, PhD thesis, The University of Ghent, 2002] that an m-system of W(4m + 3, q) or Q − (4m + 3, q) is a pseudo-ovoid of the ambient projective space.
Let S be an essentially smooth scheme over a field and ℓ = char S a prime number. We show that the algebra of bistable operations in the mod ℓ motivic cohomology of smooth S-schemes is generated by the motivic Steenrod operations. This was previously proved by Voevodsky for S a field of characteristic zero. We follow Voevodsky's proof but remove its dependence on characteristic zero by usingétale cohomology instead of topological realization and by replacing resolution of singularities with a theorem of Gabber on alterations.
BackgroundTo study the role of microRNA (miRNA) in the regulation of Chinese hamster ovary (CHO) cell growth, qPCR, microarray and quantitative LC-MS/MS analysis were utilised for simultaneous expression profiling of miRNA, mRNA and protein. The sample set under investigation consisted of clones with variable cellular growth rates derived from the same population. In addition to providing a systems level perspective on cell growth, the integration of multiple profiling datasets can facilitate the identification of non-seed miRNA targets, complement computational prediction tools and reduce false positive and false negative rates.Results51 miRNAs were associated with increased growth rate (35 miRNAs upregulated and 16 miRNAs downregulated). Gene ontology (GO) analysis of genes (n=432) and proteins (n=285) found to be differentially expressed (DE) identified biological processes driving proliferation including mRNA processing and translation. To investigate the influence of miRNA on these processes we combined the proteomic and transcriptomic data into two groups. The first set contained candidates where evidence of translational repression was observed (n=158). The second group was a mixture of proteins and mRNAs where evidence of translational repression was less clear (n=515). The TargetScan algorithm was utilised to predict potential targets within these two groups for anti-correlated DE miRNAs.ConclusionsThe evidence presented in this study indicates that biological processes such as mRNA processing and protein synthesis are correlated with growth rate in CHO cells. Through the integration of expression data from multiple levels of the biological system a number of proteins central to these processes including several hnRNPs and components of the ribosome were found to be post-transcriptionally regulated. We utilised the expression data in conjunction with in-silico tools to identify potential miRNA-mediated regulation of mRNA/proteins involved in CHO cell growth rate. These data have allowed us to prioritise candidates for cell engineering and/or biomarkers relevant to industrial cell culture. We also expect the knowledge gained from this study to be applicable to other fields investigating the role of miRNAs in mammalian cell growth.
Recently released sequence information on Chinese hamster ovary (CHO) cells promises to not only facilitate our understanding of these industrially important cell factories through direct analysis of the sequence, but also to enhance existing methodologies and allow new tools to be developed. In this article we demonstrate the utilization of CHO specific sequence information to improve mass spectrometry (MS) based proteomic identification. The use of various CHO specific databases enabled the identification of 282 additional proteins, thus increasing the total number of identified proteins by 40-50%, depending on the sample source and methods used. In addition, a considerable portion of those proteins that were identified previously based on inter-species sequence homology were now identified by a larger number of peptides matched, thus increasing the confidence of identification. The new sequence information offers improved interpretation of proteomic analyses and will, in the years to come, prove vital to unraveling the CHO proteome.
We give scheme-theoretic descriptions of the category of fibre functors on the categories of sheaves associated to the Zariski, Nisnevich, étale, rh, cdh, ldh, eh, qfh, and h topologies on the category of separated schemes of finite type over a separated noetherian base. Combined with a theorem of Deligne on the existence of enough points, this provides an algebro-geometric description of a conservative family of fibre functors on these categories of sheaves. As an example of an application we show that direct image along a closed immersion is exact for all these topologies except qfh. The methods are transportable to other categories of sheaves as well.1 The second is valid in many other situations which the reader can work out according to their needs. For example, the étale, qfh, Nisnevich, Zariski, etc topologies admit various "small" sites, and when C is taken to be such a small site instead of Sch/S, we obtain the full subcategory of the category of (C, τ )-local schemes consisting of those which are obtainable as inverse limits of pro-objects of C. 2 By abuse of notation, by f (P ) we mean lim − − → (P \Sch/S) f (X), where (P \Sch/S) is the category of factorizations P → X → S with X ∈ Sch/S.
microRNA engineering of CHO cells has already proved successful in enhancing various industrially relevant phenotypes and producing various recombinant products. A single miRNA's ability to interact with multiple mRNA targets allows their regulatory capacity to extend to processes such as cellular metabolism. Various metabolic states have previously been associated with particular CHO cell phenotypes such as glycolytic or oxidative metabolism accommodating growth and productivity, respectively. miR-23 has previously been demonstrated to play a role in glutamate metabolism resulting in enhanced oxidative phosphorylation through the TCA cycle. Re-programming cellular bioenergetics through miR-23 could tip the balance, forcing mammalian production cells to be more productive by favoring metabolic channelling into oxidative metabolism. CHO clones depleted of miR-23 using a miR-sponge decoy demonstrated an average ∼three-fold enhanced specific productivity with no impact on cell growth. Using a cell respirometer, mitochondrial activity was found to be enhanced by ∼30% at Complex I and II of the electron transport system. Additionally, label-free proteomic analysis uncovered various potential novel targets of miR-23 including LE1 and IDH1, both implicated in oxidative metabolism and mitochondrial activity. These results demonstrate miRNA-based engineering as a route to re-programming cellular metabolism resulting in increased productivity, without affecting growth.
Identification of protein targets that play a role in breast cancer invasion may help to understand the rapid progression of cancer and may lead to the development of new biomarkers for the disease. In this study, we compared two highly invasive and two poorly invasive breast cancer cell lines using comparative label-free LC-MS profiling in order to identify differentially expressed proteins that may be linked to the invasive phenotype in vitro. Forty-five proteins were found to be upregulated, and 34 proteins, downregulated. UV excision repair protein RAD23 homologue B (RAD23B) was found among the downregulated proteins in highly invasive breast cancer cell lines. In poorly invasive breast cancer cell lines, siRNA-mediated downregulation of RAD23B subsequently led to an increase in invasion and adhesion in vitro. Immunohistochemistry analysis of 164 specimens of invasive breast cancer showed that having a high percentage (>80%) of RAD23B positive nuclei was significantly associated with histopathological grades 1 and 2 breast cancer and with low mitotic activity. In addition, a high staining intensity for RAD23B in the cytoplasm was significantly associated with histopathological grade 3 breast cancer. This study suggests a potential role of RAD23B in breast cancer progression and may further imply a tumor suppressor role of nuclear RAD23B in breast cancer.
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