Use of semipermeable polyurethane hollow fibers for pituitary organ culture

Summary A new model for organ culture of endocrine tissue is described. Rat anterior pituitary fragments were cultured for 4 wk within semipermeable polyurethane isocyanate hollow fibers. Growth hormone and prolactin, two of the anterior pituitary hormones, were released into the medium during the entire culture period. Electron microscopy of the pituitary fragments after 2 wk in in culture showed a rim of viable tissue in all specimens examined. Individual cells, from this outer rim, exhibited excellent organelle preservation and numerous secretory granules. Experiments involving potassium depolarization and 10⁻⁶ M dopamine provided evidence for the normal responsiveness of the cultured pituitary tissue to both stimulatory and inhibitory factors. These studies illustrate the potential utility of the described organ culture system for further investigations of endocrine physiology.     

Evolutionary relationships of the New Caledonian heterotrophic conifer, Parasitaxus usta (Podocarpaceae), inferred from chloroplast trnL-F intron/spacer and nuclear rDNA ITS2 sequences

 The phylogenetic position of Parasitaxus (Podocarpaceae) has been inferred from a cladistic analysis of molecular characters from chloroplast and nuclear genomes including all genera of Podocarpaceae. In all 24 most parsimonious trees, based on combined datasets, Phyllocladus resided outside Podocarpaceae s. str. while Lepidothamnus was basal to the latter. Most other genera were arranged in two major clades. The evidence confirms previous studies, which have suggested a relationship between Lagarostrobos, Manoao and Parasitaxus. Parasitaxus is not directly related to its host Falcatifolium taxoides. Instead it appears to be most closely related to Manoao and Lagarostrobos. No other members of this group now occur on New Caledonia. However, if the evolution of Parasitaxus were autochthonous, a free-living member of this group must once have occurred there. An accelerated evolutionary rate of the chloroplast sequence analysed was suggested, indicating that the plant behaves like a holoparasite. Received January 4, 2002; accepted April 3, 2002 Published online: September 13, 2002     

NMR assignments for the Sinorhizobium meliloti response regulator Sma0114

Response regulators are terminal ends of bacterial two-component systems that undergo extensive structural reorganization in response to phosphoryl transfer from their cognate histidine kinases. The response regulator encoded by the gene sma0114 of Sinorhizobium meliloti is a part of a unique class of two-component systems that employ HWE histidine kinases. The distinct features of Sma0114 include a PFxFATGY motif that houses the conserved threonine in the “Y–T coupling” conformational switch which mediates output response through downstream protein–protein interactions, and the replacement of the conserved phenylalanine/tyrosine in Y–T coupling by a leucine. Here we present ¹H, ¹⁵N, and ¹³C NMR assignments for Sma0114. We identify the secondary structure of the protein based on TALOS chemical shift analysis, ³J_HNHα coupling constants and hydrogen–deuterium exchange. The secondary structure determined by NMR is in good agreement with that predicted from the sequence. Both methods suggest that Sma0114 differs from standard CheY-like folds by missing the fourth α-helix. Our initial NMR characterization of Sma0114 paves the way to a full investigation of the structure and dynamics of this response regulator.     

Comparison of genome diversity of Brucella spp. field isolates using Universal Bio-signature Detection Array and whole genome sequencing reveals limitations of current diagnostic methods

The detection and identification of bio-threat agents and the study of host-pathogen interactions require a high-resolution detection platform capable of discerning closely related species. Diverse analysis methods are used to identify pathogens, specifically Brucella species or biovars. In this study, we compared four diagnostic approaches including serology-based biochemical test, PCR assay, microarray analysis using a Universal Bio-signature Detection Array (UBDA) and whole genome "deep" sequencing for Brucella organisms including a number of field isolates. We found that although there was frequent agreement among the different tests, some tests gave compound/contradictory results that were a consequence of species diversity due to mixed infections or minor contaminants as measured by UBDA and validated from whole genome sequence. By comparing these analysis techniques, we demonstrate that standard diagnostics used in the field are limited in their ability to identify genomic DNA contaminants in field isolates while UBDA and sequencing analysis are highly sensitive in tracing genomic differences among the isolates.     

First epigravettian ceramic figurines from europe (vela spila, croatia)

Recent finds of 36 ceramic artifacts from the archaeological site of Vela Spila, Croatia, offer the first evidence of ceramic figurative art in late Upper Palaeolithic Europe, c. 17,500-15,000 years before present (BP). The size and diversity of this artistic ceramic assemblage indicate the emergence of a social tradition, rather than more ephemeral experimentation with a new material. Vela Spila ceramics offer compelling technological and stylistic comparisons with the only other evidence of a developed Palaeolithic ceramic tradition found at the sites of Pavlov I and Dolní Věstonice I, in the Czech Republic, c. 31,000-27,000 cal BP. Because of the 10,000-year gap between the two assemblages, the Vela Spila ceramics are interpreted as evidence of an independent invention of this technology. Consequently, these artifacts provide evidence of a new social context in which ceramics developed and were used to make art in the Upper Palaeolithic.     

Serotonylated fibronectin is elevated in pulmonary hypertension

Serotonin (5-HT) and fibronectin (FN) have been associated with pulmonary hypertension (PH). We previously reported that FN is posttranslationally modified by tissue transglutaminase (TGase) to form serotonylated FN (s-FN) in pulmonary artery smooth muscle cells and that serotonylation stimulates their proliferation and migration, hallmarks of PH. We hypothesized that s-FN and its binding to TGase are elevated in human and experimental PH. To assess this hypothesis, FN isolation and electrophoretic, immunoblotting, and densitometric techniques were used. Mean ratio of serum s-FN to total FN level (s-FN/FN) was elevated in 19 consecutive pulmonary arterial hypertension (PAH) patients compared with 25 controls (0.3 ± 0.18 vs. 0.05 ± 0.07, P < 0.001). s-FN/FN also was increased in lungs of mice and rats with hypoxia-induced PH and in rats with monocrotaline-induced PH. In mice, the increase was detected at 1 wk of hypoxia, preceding the development of PH. Hypoxic rats had elevated serum s-FN/FN. Enhanced binding of TGase to its substrate FN occurred in serum from patients with PAH (mean 0.50 ± 0.51 vs. 0.063 ± 0.11, P = 0.002) and s-FN/FN and TGase-bound FN were highly correlated (R(2) = 0.77). TGase-bound FN also was increased in experimental PH. We conclude that increased serotonylation of FN occurs in human and experimental PH and may provide a biomarker for the disease.