Fatal bilateral chylothorax in mice lacking the integrin alpha9beta1

Members of the integrin family of adhesion receptors mediate both cell-cell and cell-matrix interactions and have been shown to play vital roles in embryonic development, wound healing, metastasis, and other biological processes. The integrin alpha9beta1 is a receptor for the extracellular matrix proteins osteopontin and tenacsin C and the cell surface immunoglobulin vascular cell adhesion molecule-1. This receptor is widely expressed in smooth muscle, hepatocytes, and some epithelia. To examine the in vivo function of alpha9beta1, we have generated mice lacking expression of the alpha9 subunit. Mice homozygous for a null mutation in the alpha9 subunit gene appear normal at birth but develop respiratory failure and die between 6 and 12 days of age. The respiratory failure is caused by an accumulation of large volumes of pleural fluid which is rich in triglyceride, cholesterol, and lymphocytes. alpha9(-/-) mice also develop edema and lymphocytic infiltration in the chest wall that appears to originate around lymphatics. alpha9 protein is transiently expressed in the developing thoracic duct at embryonic day 14, but expression is rapidly lost during later stages of development. Our results suggest that the alpha9 integrin is required for the normal development of the lymphatic system, including the thoracic duct, and that alpha9 deficiency could be one cause of congenital chylothorax.     

Plasmin-sensitive dibasic sequences in the third fibronectin-like domain of L1-cell adhesion molecule (CAM) facilitate homomultimerization and concomitant integrin recruitment

L1 is a multidomain transmembrane neural recognition molecule essential for neurohistogenesis. While moieties in the immunoglobulin-like domains of L1 have been implicated in both heterophilic and homophilic binding, the function of the fibronectin (FN)-like repeats remains largely unresolved. Here, we demonstrate that the third FN-like repeat of L1 (FN3) spontaneously homomultimerizes to form trimeric and higher order complexes. Remarkably, these complexes support direct RGD-independent interactions with several integrins, including alpha(v)beta(3) and alpha(5)beta(1). A pep- tide derived from the putative C-C' loop of FN3 (GSQRKHSKRHIHKDHV(852)) also forms trimeric complexes and supports alpha(v)beta(3) and alpha(5)beta(1) binding. Substitution of the dibasic RK(841) and KR(845) sequences within this peptide or the FN3 domain limited multimerization and abrogated integrin binding. Evidence is presented that the multimerization of, and integrin binding to, the FN3 domain is regulated both by conformational constraints imposed by other domains and by plasmin- mediated cleavage within the sequence RK( downward arrow)HSK( downward arrow)RH(846). The integrin alpha(9)beta(1), which also recognizes the FN3 domain, colocalizes with L1 in a manner restricted to sites of cell-cell contact. We propose that distal receptor ligation events at the cell-cell interface may induce a conformational change within the L1 ectodomain that culminates in receptor multimerization and integrin recruitment via interaction with the FN3 domain.     

Effect of patch size and plant density of Paterson"s curse (Echium plantagineum) on the oviposition of a specialist weevil, Mogulones larvatus

This study examines the oviposition response of a specialist weevil (Mogulones larvatus) to patches of its host, the noxious weed Paterson"s curse/salvation Jane (Echium plantagineum). We simultaneously examined the effect of patch size and plant density (and their interaction), on the recorded oviposition patterns. Our results show that oviposition first occurred on the largest patches with the highest number of plants. However, there was no significant effect of patch size or number of plants per patch at the end of the experiment. At this time the level of attack per plant was negatively correlated with plant density. This negative effect of density on the level of oviposition was not mediated by a reduction in plant size at higher densities. The pattern observed may indicate a risk-spreading strategy by females. Received: 15 July 1999 / Accepted: 14 April 2000     

Biodiversity patterns in Indian Ocean corals, and effects of taxonomic error in data

Corals from 26 sites in the Indian Ocean, from numerous taxonomic sources, are analysed for distribution patterns after applying a consistent synonymy. The data set contains double the amount of distribution information used previously. Cluster analysis shows several contiguous regional groupings within this ocean, and no geographically dispersed groupings exist. Between-site species similarity correlates negatively with distance between sites. Coral species diversity and genus diversity plotted with latitude and longitude confirm that a band of high diversity stretches across the entire Indian Ocean, but that in this Ocean, unlike the Pacific and greater Caribbean area, there is no clear gradient with latitude at least up to the latitudinal limits of coral growth. About half the species are widespread throughout the Indian Ocean. Species frequency/distribution curves and cumulative frequency curves show that 150 species occur at only 2–4 sites. Multi-dimensional scaling (MDS) analysis using only these species confirms that they are important in forming sub-regional groupings which are superimposed on a general Indian Ocean homogeneity.The perennial problem of error in taxonomic data sets is examined. Increasing taxonomic error is introduced into the data using random methods. It is found that about 25% more error can be added to this data set before the relationship between similarity coefficient and geographical distance between pairs of sites is lost. Measures extracted from the clustering procedure using the original data and the data sets with added taxonomic error, show a sharp loss in cluster formation after addition of about 10 or 20% more error.     

Structure of an archaeal non-discriminating glutamyl-tRNA synthetase: a missing link in the evolution of Gln-tRNAGln formation

The molecular basis of the genetic code relies on the specific ligation of amino acids to their cognate tRNA molecules. However, two pathways exist for the formation of Gln-tRNA^Gln. The evolutionarily older indirect route utilizes a non-discriminating glutamyl-tRNA synthetase (ND-GluRS) that can form both Glu-tRNA^Glu and Glu-tRNA^Gln. The Glu-tRNA^Gln is then converted to Gln-tRNA^Gln by an amidotransferase. Since the well-characterized bacterial ND-GluRS enzymes recognize tRNA^Glu and tRNA^Gln with an unrelated α-helical cage domain in contrast to the β-barrel anticodon-binding domain in archaeal and eukaryotic GluRSs, the mode of tRNA^Glu/tRNA^Gln discrimination in archaea and eukaryotes was unknown. Here, we present the crystal structure of the Methanothermobacter thermautotrophicus ND-GluRS, which is the evolutionary predecessor of both the glutaminyl-tRNA synthetase (GlnRS) and the eukaryotic discriminating GluRS. Comparison with the previously solved structure of the Escherichia coli GlnRS-tRNA^Gln complex reveals the structural determinants responsible for specific tRNA^Gln recognition by GlnRS compared to promiscuous recognition of both tRNAs by the ND-GluRS. The structure also shows the amino acid recognition pocket of GluRS is more variable than that found in GlnRS. Phylogenetic analysis is used to reconstruct the key events in the evolution from indirect to direct genetic encoding of glutamine.     

Adverse implications of misdating in dendrochronology: Addressing the re-dating of the “Messiah” violin

A recent report by Mondino and Avalle (2009) was widely distributed that demonstrated a re-dating of the famous “Messiah” violin, a violin attributed to Antonio Stradivari with a label date of 1716. An outermost ring date of 1844 is instead suggested rather than dates in the 1680s reported in previous studies. Mondino and Avalle suggest that this outermost ring date supports the attribution of the violin to Jean-Baptiste Vuillaume, a prolific French instrument maker who was well known for his copies of famous instruments. The statistical techniques and exercises used by Mondino and Avalle, however, are problematic and do not support this revised outermost date for the “Messiah” violin. Raw measurement data with original trends are used in their statistical crossdating, properties previously shown to hinder precise crossdating. They then substantiate their re-dating with polynomial trend curves, which has ever been accepted practice in dendrochronology. Furthermore, the authors use re-scaled correlation coefficients and t-values which artificially inflate the strength of the relationship between tree-ring series that are being statistically crossdated. Using the exact same tree-ring data, but using accepted techniques in statistical crossdating (e.g., the removal of all low-frequency trends and autocorrelation), we could not verify the revised dating of the “Messiah” violin. We urge caution for those who intend to use the SynchroSearch software, book, and lesson plans developed and distributed by Mondino and Avalle. This study illustrates the adverse effects possible in dendrochronology when investigators do not adhere to accepted and time-tested techniques, and are not versed in the extensive literature that highlights issues commonly encountered in statistical crossdating.     

Factors that influence the radiofrequency power output of GSM mobile phones

Epidemiological studies of mobile phone use and risk of brain cancer have relied on self-reported use, years as a subscriber, and billing records as exposure surrogates without addressing the level of radiofrequency (RF) power output. The objective of this study was to measure environmental, behavioral and engineering factors affecting the RF power output of GSM mobile phones during operation. We estimated the RF-field exposure of volunteer subjects who made mobile phone calls using software-modified phones (SMPs) that recorded output power settings. Subjects recruited from three geographic areas in the U.S. were instructed to log information (place, time, etc.) for each call made and received during a 5-day period. The largest factor affecting energy output was study area, followed by user movement and location (inside or outside), use of a hands-free device, and urbanicity, although the two latter factors accounted for trivial parts of overall variance. Although some highly statistically significant differences were identified, the effects on average energy output rate were usually less than 50% and were generally comparable to the standard deviation. These results provide information applicable to improving the precision of exposure metrics for epidemiological studies of GSM mobile phones and may have broader application for other mobile phone systems and geographic locations.     

Addressing childhood overweight through schools

Rates of childhood obesity in have reached alarming proportions in many countries. Sixteen percent of school-aged children and adolescents in the U.S. are overweight. Legislation implemented in 2004 in the U.S. requires local education agencies (LEAs) that sponsor school meal programs to establish local wellness policies to address childhood obesity. Project PA, a collaboration between a state agency and a university providing school-based interventions focuses on the school environment and policy changes. Interventions have targeted foodservice personnel, administrators, teachers, parents and students. In two recent projects schools assessed their school nutrition environments, developed nutrition policies, and implemented strategies to encourage healthier food selections. Schools identified weaknesses in the areas of marketing and communication of policies. Media attention on the childhood obesity facilitated policy changes. Time and cost were identified as barriers to policy development and there were concerns about weak enforcement of policies. These themes are discussed.     

Candida albicans protein kinase CK2 governs virulence during oropharyngeal candidiasis

To identify Candida albicans genes whose proteins are necessary for host cell interactions and virulence, a collection of C. albicans insertion mutants was screened for strains with reduced capacity to damage endothelial cells in vitro. This screen identified CKA2. CKA2 and its homologue CKA1 encode the catalytic subunits of the protein kinase CK2. cka2delta/cka2delta strains of C. albicans were constructed and found to have significantly reduced capacity to damage both endothelial cells and an oral epithelial cell line in vitro. Although these strains invaded endothelial cells similarly to the wild-type strain, they were defective in oral epithelial cell invasion. They were also hypersusceptible to hydrogen peroxide, but not to high salt or to cell wall damaging agents. A cka1delta/cka1delta mutant caused normal damage to both endothelial cells and oral epithelial cells, and it was not hypersusceptible to hydrogen peroxide. However, overexpression of CKA1 in a cka2delta/cka2delta strain restored wild-type phenotype. Although the cka2delta/cka2delta mutant had normal virulence in the mouse model of haematogenously disseminated candidiasis, it had significantly attenuated virulence in the mouse model of oropharyngeal candidiasis. Therefore, Cka2p governs the interactions of C. albicans with endothelial and oral epithelial cells in vitro and virulence during oropharyngeal candidiasis.