The thyroid hormone receptor functions as a ligand-operated developmental switch between proliferation and differentiation of erythroid progenitors

The avian erythroblastosis virus (AEV) oncoprotein v-ErbA represents a mutated, oncogenic thyroid hormone receptor alpha (c-ErbA/ TRalpha). v-ErbA cooperates with the stem cell factor-activated, endogenous receptor tyrosine kinase c-Kit to induce self-renewal and to arrest differentiation of primary avian erythroblasts, the AEV transformation target cells. In this cooperation, v-ErbA substitutes for endogenous steroid hormone receptor function required for sustained proliferation of non-transformed erythroid progenitors. In this paper, we propose a novel concept of how v-ErbA transforms erythroblasts. Using culture media strictly depleted from thyroid hormone (T3) and retinoids, the ligands for c-ErbA/TRalpha and its co-receptor RXR, we show that overexpressed, unliganded c-ErbA/ TRalpha closely resembles v-ErbA in its activity on primary erythroblasts. In cooperation with ligand-activated c-Kit, c-ErbA/ TRalpha causes steroid-independent, long-term proliferation and tightly blocks differentiation. Activation of c-ErbA/ TRalpha by physiological T3 levels causes the loss of self-renewal capacity and induces synchronous, terminal differentiation under otherwise identical conditions. This T3-induced switch in erythroid progenitor development is correlated with a decrease of c-ErbA-associated histone deacetylase activity. Our results suggest that the crucial role of the mutations activating v-erbA as an oncogene is to 'freeze' c-ErbA/ TRalpha in its non-liganded, repressive conformation and to facilitate its overexpression.     

Use of a Mycobacterium tuberculosis H37Rv bacterial artificial chromosome library for genome mapping, sequencing, and comparative genomics

The bacterial artificial chromosome (BAC) cloning system is capable of stably propagating large, complex DNA inserts in Escherichia coli. As part of the Mycobacterium tuberculosis H37Rv genome sequencing project, a BAC library was constructed in the pBeloBAC11 vector and used for genome mapping, confirmation of sequence assembly, and sequencing. The library contains about 5,000 BAC clones, with inserts ranging in size from 25 to 104 kb, representing theoretically a 70-fold coverage of the M. tuberculosis genome (4.4 Mb). A total of 840 sequences from the T7 and SP6 termini of 420 BACs were determined and compared to those of a partial genomic database. These sequences showed excellent correlation between the estimated sizes and positions of the BAC clones and the sizes and positions of previously sequenced cosmids and the resulting contigs. Many BAC clones represent linking clones between sequenced cosmids, allowing full coverage of the H37Rv chromosome, and they are now being shotgun sequenced in the framework of the H37Rv sequencing project. Also, no chimeric, deleted, or rearranged BAC clones were detected, which was of major importance for the correct mapping and assembly of the H37Rv sequence. The minimal overlapping set contains 68 unique BAC clones and spans the whole H37Rv chromosome with the exception of a single gap of approximately 150 kb. As a postgenomic application, the canonical BAC set was used in a comparative study to reveal chromosomal polymorphisms between M. tuberculosis, M. bovis, and M. bovis BCG Pasteur, and a novel 12.7-kb segment present in M. tuberculosis but absent from M. bovis and M. bovis BCG was characterized. This region contains a set of genes whose products show low similarity to proteins involved in polysaccharide biosynthesis. The H37Rv BAC library therefore provides us with a powerful tool both for the generation and confirmation of sequence data as well as for comparative genomics and other postgenomic applications. It represents a major resource for present and future M. tuberculosis research projects.     

Purification and characterization of a high molecular weight histone deacetylase complex (HD2) of maize embryos

The dynamic state of core histone acetylation is maintained by histone acetyltransferases and deacetylases. In germinating maize embryos, four nuclear histone deacetylases can be distinguished. From a chromatin fraction prepared at 72 h after start of embryo germination, we have purified the nuclear histone deacetylase HD2 to homogeneity. Using a sequence of chromatographic steps, we achieved the purification of an enzymatically active high molecular weight protein complex with an apparent molecular mass of 400 kDa, as determined by gel filtration chromatography. The purified enzyme was characterized in terms of enzymatic and kinetic properties, and sensitivity to several histone deacetylase inhibitors. In SDS-polyacrylamide gels, HD2 split into three polypeptides of 45, 42, and 39 kDa, suggesting that the native enzyme is a multimer-protein complex. Electrophoresis under nondenaturing conditions in combination with second dimension SDS-gel electrophoresis indicated that all three protein components of the HD2 complex were enzymatically active. Polyclonal antibodies against each of the three polypeptides were raised in rabbits. Each antiserum reacted with all three polypeptides on Western blots, suggesting that p45, p42, and p39 are highly homologous. This homology was confirmed by amino acid sequencing of peptides generated from each of the three HD2 components.     

Geology and the classification of rock masses—Examples from Austrian tunnels

Independent of the well known classification systems this paper presents an attempt at a subsequent, geological characterisation of the classes of rock mass quality found for three tunnel projects in Austria. Simple statistical treatment of the data shows that joint and bedding-plane spacing, sliding prone fillings and seepage are the most useful geological criteria for characterising the rock mass classes, which themselves had been derived from standard support requirements. Ratio and relative importance of these criteria, however, are quite different for each tunnel project. Influence of tunnel cross sectional area, depth of cover and orientation of divisional planes could not be proved important on the basis of the data available.     

Aroyl-pyrrolyl hydroxyamides: influence of pyrrole C4-phenylacetyl substitution on histone deacetylase inhibition

The novel aroyl-pyrrolyl hydroxyamides 4 a-a' are analogues of the lead compound 3-(1-methyl-4-phenylacetyl-1H-pyrrol-2-yl)-N-hydroxy-2-propenamide (2) and are active as HDAC inhibitors. The benzene ring of 2 was substituted with a wide range of electron-donating and electron-withdrawing groups, and the effect was evaluated on three HDACs from maize, namely HD2, HD1-B (a class I HDAC), and HD1-A (a class II HDAC). Inhibition studies show that the benzene 3' and, to a lesser extent, 4' positions of 2 were the most suitable for the introduction of substituents, with the 3'-chloro (in 4 b) and the 3'-methyl (in 4 k) derivatives being the most potent compounds, reaching the same activity as SAHA. Inhibition data for 4 b,k against mouse HDAC1 were consistent with those observed in the maize enzyme. The substituent insertion on the benzene ring of 2 (compounds 4 a-a') abated the slight (3-fold) selectivity for class II HDACs displayed by 2. Compound 4 b showed interesting, dose-dependent antiproliferative and cytodifferentiation properties against human acute promyelocytic leukemia HL-60 cells.     

Use of pulsed field gel electrophoresis to compare large DNA-restriction fragments of Listeria monocytogenes strains belonging to serogroups 1/2 and 3.

Thirty-five Listeria monocytogenes strains belonging to serogroups 1/2 and 3 and isolated from various origins were characterized by whole cellular DNA restriction patterns using low-frequency cleavage enzymes and pulsed field gel electrophoresis. Seventeen restriction profiles were detected with ApaI, 18 with SmaI and 15 with NotI, the combination of these patterns allowing one to define at least 24 distinct groups within the 35 strains. The significant genomic diversity pointed out by this method can be of value in the epidemiological fingerprinting of L. monocytogenes.     

The high-pathogenicity island of Yersinia pseudotuberculosis can be inserted into any of the three chromosomal asn tRNA genes

In two experiments, we investigated how reading time was affected by the plausibility of the prepositional phrase in subject-verb-noun-phrase-prepositional-phrase sentences, and the status of the prepositional phrase as argument versus adjunct of the verb. Highly plausible prepositional phrases were read faster than less plausible ones, and argument prepositional phrases were read faster than adjuncts. These effects appeared both in a self-paced reading experiment and in an experiment that measured eye movements during normal reading. The effects of plausibility were substantially larger and longer lasting than the effects of argument status, but both appeared very early in the reading of the prepositional phrase. The implications of these effects for models of parsing and sentence interpretation are discussed.     

The significance and prediction of different rock mass characteristics for rock engineering

This paper introduces a methodology to quantify the significance of changes in certain rock mass characteristics. The developed method can be applied to all data sets that consist of parameters classified into categories. The tools are developed from the principles of engineering geological rock mass characterisation and statistical theory. The basic approach is the construction of the so-called length matrix, which allows for the combination of observed changes in parameter categories and their change in lengths in a sequence of observations. The concept of the order of characteristic changes is introduced to construct the length matrix of two or more combined categories, which is essential to assess the significance of different rock mass characteristic categories, their consistency and persistence. By using the mathematical formulae introduced in this paper, four experimental models can be produced for engineering purposes. The first model is the frequency matrix, which gives the change length frequencies between different categories and is only valid in the range of observed data. The second model is the probability matrix, which describes the generic length probability of significant rock mass characteristic categories. The third model is the transition probability matrix, which predicts the generic change dependencies of significant categories. The last model, the model of multiple sequences, identifies probabilistically the two- or three-dimensional change consistencies of significant rock mass characteristic categories. The data sets of fracture intensity categories in the flat-lying limestone series of Birecik dam bedrock (Turkey) were used for verification and demonstration of applications. The first example shows the identification of the significant patterns of fracture intensity categories by applying the probability matrix. The model was verified in the second example and the patterns of fracture intensity categories were reliably predicted by applying a transition probability matrix. In the last example, the fracture intensity categories of dam bedrock were partitioned by applying the model of multiple sequences.     

The Role of Fear-Relevant Stimuli in Visual Search: A Comparison of Phylogenetic and Ontogenetic Stimuli.

It has been argued that phylogenetic fear-relevant stimuli elicit preattentive capture of attention. To distinguish between fear relevance and time of appearance in evolutionary history, the authors compare phylogenetic and ontogenetic fear-relevant and fear-irrelevant stimuli in a visual search task. The authors found no evidence for a special role of phylogenetic fear-relevant stimuli; it seems that fear relevance in general is more important than is the evolutionary age. The pattern of results indicates that attention toward threatening stimuli is mainly affected by a late component that prolongs the disengagement of attention. (PsycINFO Database Record (c) 2010 APA, all rights reserved)