Immunization with Dendritic Cells Pulsed ex vivo with Recombinant Chlamydial Protease-Like Activity Factor Induces Protective Immunity Against Genital Chlamydia muridarum Challenge

We have shown that immunization with soluble recombinant chlamydial protease-like activity factor (rCPAF) and a T helper 1 type adjuvant can induce significantly enhanced bacterial clearance and protection against Chlamydia-induced pathological sequelae in the genital tract. In this study, we investigated the use of bone marrow derived dendritic cells (BMDCs) pulsed ex vivo with rCPAF + CpG in an adoptive subcutaneous immunization for the ability to induce protective immunity against genital chlamydial infection. We found that BMDCs pulsed with rCPAF + CpG efficiently up-regulated the expression of activation markers CD86, CD80, CD40, and major histocompatibility complex class II (MHC II), and secreted interleukin-12, but not IL-10 and IL-4. Mice adoptively immunized with rCPAF + CpG-pulsed BMDCs or UV-EB + CpG-pulsed BMDCs produced elevated levels of antigen-specific IFN-γ and enhanced IgG1 and IgG2a antibodies. Moreover, mice immunized with rCPAF + CpG-pulsed BMDCs or UV-EB + CpG-pulsed BMDCs exhibited significantly reduced genital Chlamydia shedding, accelerated resolution of infection, and reduced oviduct pathology when compared to infected mock-immunized animals. These results suggest that adoptive subcutaneous immunization with ex vivo rCPAF-pulsed BMDCs is an effective approach, comparable to that induced by UV-EB–BMDCs, for inducing robust anti-Chlamydia immunity

Beginner\u27s luck: a language for property-based generators

Property-based random testing a la QuickCheck requires building efficient generators for well-distributed random data satisfying complex logical predicates, but writing these generators can be difficult and error prone. We propose a domain-specific language in which generators are conveniently expressed by decorating predicates with lightweight annotations to control both the distribution of generated values and the amount of constraint solving that happens before each variable is instantiated. This language, called Luck, makes generators easier to write, read, and maintain. We give Luck a formal semantics and prove several fundamental properties, including the soundness and completeness of random generation with respect to a standard predicate semantics. We evaluate Luck on common examples from the property-based testing literature and on two significant case studies, showing that it can be used in complex domains with comparable bug-finding effectiveness and a significant reduction in testing code size compared to handwritten generators

Efficiency of generalized estimating equations for binary responses

Using standard correlation bounds, we show that in generalized estimation equations (GEEs) the so-called 'working correlation matrix' R("&agr;") for analysing binary data cannot in general be the true correlation matrix of the data. Methods for estimating the correlation param-eter in current GEE software for binary responses disregard these bounds. To show that the GEE applied on binary data has high efficiency, we use a multivariate binary model so that the covariance matrix from estimating equation theory can be compared with the inverse Fisher information matrix. But R ("&agr;") should be viewed as the weight matrix, and it should not be confused with the correlation matrix of the binary responses. We also do a comparison with more general weighted estimating equations by using a matrix Cauchy-Schwarz inequality. Our analysis leads to simple rules for the choice of "&agr;" in an exchangeable or autoregressive AR(1) weight matrix R ("&agr;"), based on the strength of dependence between the binary variables. An example is given to illustrate the assessment of dependence and choice of "&agr;". Copyright 2004 Royal Statistical Society.

"Clicktophycin-52": A Bioactive Cryptophycin-52 Triazole Analogue

Nahrwold M, Bogner T, Eissler S, Verma S, Sewald N. "Clicktophycin-52": A Bioactive Cryptophycin-52 Triazole Analogue. Organic Letters. 2010;12(5):1064-1067.An endocyclic trans-amide linkage within the macrocyclic antitumor agent cryptophycin-52 was replaced by a 1,4-disubstituted 1H-1,2,3-triazole ring. Macrocyclisation of the triazole analogue was accomplished by macrolactamization as well as by Cu(I)-mediated "click"-cyclization. Compared to cryptophycin-52, in vitro cytotoxicity of "clicktophycin-52" against the multidrug resistant human cancer cell line KB-V1 is only slightly reduced