In vitro differentiation of near-unlimited numbers of functional mouse basophils using conditional Hoxb8

Background: Basophils constitute a rare leukocyte population known for their effector functions in inflammation and allergy, as well as more recently described immunoregulatory roles. Besides their low frequency, functional analysis of basophils is hindered by a short life span, inefficient ex vivo differentiation protocols, and lack of suitable cell models. A method to produce large quantities of basophils in vitro would facilitate basophil research and constitute a sought-after tool for diagnostic and drug testing purposes. Methods: A method is described to massively expand bone marrow–derived basophils in vitro. Myeloid progenitors are conditionally immortalized using Hoxb8 in the presence of interleukin-3 (IL-3) and outgrowing cell lines selected for their potential to differentiate into basophils upon shutdown of Hoxb8 expression. Results: IL-3-dependent, conditional Hoxb8-immortalized progenitor cell lines can be expanded and maintained in culture for prolonged periods. Upon shutdown of Hoxb8 expression, near-unlimited numbers of mature functional basophils can be differentiated in vitro within six days. The cells are end-differentiated and short-lived and express basophil-specific surface markers and proteases. Upon IgE- as well as C5a-mediated activation, differentiated basophils release granule enzymes and histamine and secrete Th2-type cytokines (IL-4, IL-13) and leukotriene C4. IL-3-deprivation induces apoptosis correlating with upregulation of the BH3-only proteins BCL-2-interacting mediator of cell death (BIM) and p53 upregulated modulator of apoptosis (PUMA) and downregulation of proviral integration site for Moloney murine leukemia virus 1 kinase (PIM-1). Conclusion: A novel method is presented to generate quantitative amounts of mouse basophils in vitro, which moreover allows genetic manipulation of conditionally immortalized progenitors. This approach may represent a useful alternative method to isolating primary basophils.

Canonical proof nets for classical logic

Proof nets provide abstract counterparts to sequent proofs modulo rule permutations; the idea being that if two proofs have the same underlying proof-net, they are in essence the same proof. Providing a convincing proof-net counterpart to proofs in the classical sequent calculus is thus an important step in understanding classical sequent calculus proofs. By convincing, we mean that (a) there should be a canonical function from sequent proofs to proof nets, (b) it should be possible to check the correctness of a net in polynomial time, (c) every correct net should be obtainable from a sequent calculus proof, and (d) there should be a cut-elimination procedure which preserves correctness. Previous attempts to give proof-net-like objects for propositional classical logic have failed at least one of the above conditions. In Richard McKinley (2010) [22], the author presented a calculus of proof nets (expansion nets) satisfying (a) and (b); the paper defined a sequent calculus corresponding to expansion nets but gave no explicit demonstration of (c). That sequent calculus, called LK∗ in this paper, is a novel one-sided sequent calculus with both additively and multiplicatively formulated disjunction rules. In this paper (a self-contained extended version of Richard McKinley (2010) [22]), we give a full proof of (c) for expansion nets with respect to LK∗, and in addition give a cut-elimination procedure internal to expansion nets – this makes expansion nets the first notion of proof-net for classical logic satisfying all four criteria.

Expectation Conditional Maximization-Based Deformable Shape Registration

This paper addresses the issue of matching statistical and non-rigid shapes, and introduces an Expectation Conditional Maximization-based deformable shape registration (ECM-DSR) algorithm. Similar to previous works, we cast the statistical and non-rigid shape registration problem into a missing data framework and handle the unknown correspondences with Gaussian Mixture Models (GMM). The registration problem is then solved by fitting the GMM centroids to the data. But unlike previous works where equal isotropic covariances are used, our new algorithm uses heteroscedastic covariances whose values are iteratively estimated from the data. A previously introduced virtual observation concept is adopted here to simplify the estimation of the registration parameters. Based on this concept, we derive closed-form solutions to estimate parameters for statistical or non-rigid shape registrations in each iteration. Our experiments conducted on synthesized and real data demonstrate that the ECM-DSR algorithm has various advantages over existing algorithms.

Proof internalization in generalized Frege systems for classical logic

We present a general method for inserting proofs in Frege systems for classical logic that produces systems that can internalize their own proofs.

Censors for Boolean Description Logic

Protecting different kinds of information has become an important area of research. One aspect is to provide effective means to avoid that secrets can be deduced from the answers of legitimate queries. In the context of atomic propositional databases several methods have been developed to achieve this goal. However, in those databases it is not possible to formalize structural information. Also they are quite restrictive with respect to the specification of secrets. In this paper we extend those methods to match the much greater expressive power of Boolean description logics. In addition to the formal framework, we provide a discussion of various kinds of censors and establish different levels of security they can provide.