REF-ereeing the cytoplasmic fate of mRNA via nuclear export

The C. elegans sex-determining gene tra-2 is subject to multiple forms of regulation. A report in the June 4 issue of Molecular Cell now shows that proteins associated with the tra-2 mRNA determine its pathway of nuclear export and influence its cytoplasmic fate. These findings demonstrate an additional level of control and link nuclear export to the regulation of sexual development.     

The two-component signal transduction protein Chk1p regulates quorum sensing in Candida albicans

Regulation of hyphal morphogenesis in Candida albicans can occur through quorum sensing (QS). A QS signal, farnesol, is produced during high-density growth and inhibits morphogenesis. However, the signal transduction pathway that regulates QS is unknown. Here, we show that a C. albicans mutant lacking Chk1p but not either the Sln1p or the Nik1p histidine kinase is refractory to the inhibitory effect of farnesol both in cell suspension and during the formation of a biofilm. This study is the first to demonstrate a role for a two-component signal transduction protein in QS by a eukaryotic organism.     

Construction of a bacterial artificial chromosome library, determination of genome size, and characterization of an Hsp70 gene family in Phytophthora nicotianae

The oomycete plant pathogen Phytophthora nicotianae causes diseases on a wide range of plant species. To facilitate isolation and functional characterization of pathogenicity genes, we have constructed a large-insert bacterial artificial chromosome (BAC) library using nuclear DNA from P. nicotianae H1111. The library contains 10,752 clones with an average insert size of 90 kb and is free of mitochondrial DNA. The quality of the library was verified by hybridization with 37 genes, all of which resulted in the identification of multiple positive clones. The library is estimated to be 10.6 haploid genome equivalents based on hybridization of 23 single-copy genes and the genome size of P. nicotianae was estimated to be 95.5 Mb. Hybridization with a nuclear repetitive DNA probe revealed that 4.4% of clones in the library contained 28S rDNA. Hybridization of total genomic DNA to the library indicated that at least 39% of the BAC library contains repetitive DNA sequences. A BAC pooling strategy was developed for efficient library screening. The library was used to identify and characterize BAC clones containing an Hsp70 gene family whose four members were identified to be clustered within approximately 18 kb in the P. nicotianae genome based on the physical mapping of eight BACs spanning a genomic region of approximately 186 kb. The BAC library created provides an invaluable resource for the isolation of P. nicotianae genes and for comparative genomics studies.     

Nitric oxide synthase inhibitors have opposite effects on acute inflammation depending on their route of administration

The bulk of published data has shown that NO is proinflammatory. However, there also exists the conflicting notion that NO may be protective during an inflammatory insult. In an attempt to resolve this issue, we have compared the effects on inflammation of a range of NO synthase (NOS) inhibitors given either directly to the site of the inflammatory lesion or systemically. It was found that in the carrageenin-induced pleurisy, a single intrapleural injection of the selective inducible NO inhibitors S-(2-aminoethyl) isothiourea (AE-ITU; 3 and 10 mg/kg) and N-(3-(aminomethyl)-benzyl) acetamidine (1400W; 10 mg/kg) or the selective endothelial cell NOS inhibitor L-N(5)(1-iminoethyl)-ornithine (10 mg/kg) not only exacerbated inflammation at the very early stages of the lesion (1-6 h), but also prevented inflammatory resolution. By contrast, administering NOS inhibitors systemically ameliorated the severity of inflammation throughout the reaction. To elucidate the mechanisms by which inhibition of NO synthesis locally worsened inflammation, we found an increase in histamine, cytokine-induced neutrophil chemoattractant, superoxide, and leukotriene B(4) levels at the inflammatory site. In conclusion, this work shows that the local production of NO is protective by virtue of its ability to regulate the release of typical proinflammatory mediators and, importantly, that NOS inhibitors have differential anti-inflammatory effects depending on their route of administration.     

Resistance of perch eggs to attack by aquatic fungi

In the distinctive gelatinous Perca fluviatilis egg mass, limited fungal growth by Aphanomyces and Saprolegnia spp. especially S. diclina , occurred within dead eggs but did not spread to adjacent live eggs. Perch eggs exposed to parasitic challenge by Saprolegnia parasitica , S. dieclina (type III) and S. ferax , under fluctuating temperature regimes replicating spring water temperatures, did not have significantly greater mortality than did unchallenged controls. The observations suggest that perch eggs have some anti-fungal properties which usually prevent the spread of fungus throughout the egg mass and that under normal spring temperatures there should be negligible ecological consequences of fungal infection in perch egg masses.     

Improvement of the ecological water qualification system of rivers based on the first results of the Hungarian phytobenthos surveillance monitoring

Results of an ecological quality ratio-based qualification system, developed on the basis of the analysis of 1,161 benthic diatom dataset of the Hungarian national database, are presented herein. Using Kohonen’s Self Organising Map technique, the 25 Hungarian physiographic river types were pooled into six larger distinct categories (diatom river groups). Diatom metrics were tested for their sensitivity to the targeted stressors (nutrients, COD hydromorphological alteration) in each group. The strongest relationships were found in the case of the IPS, SI and TI indices; therefore the average of these metrics (IPSITI) was proposed as a national multimetric index for Hungarian streams. Based on IPSITI values, the ratio of moderate to worse quality water was the highest in those groups containing small rivers. In the case of large, lowland and mid-altitude rivers with fine sediment, the good and moderate ecological status was more characteristic. Applicability of the IPSITI seems to be very useful in case of small- and medium-sized rivers. For these rivers, the index showed a significant relationship with nutrients and organic pollutants. In the case of very large rivers, the stressor–index relationships were not significant because of the insufficient number of samples and the small range of stressors.     

Identification versus counting protocols as sources of uncertainty in diatom-based ecological status assessments

The heterogeneity of oxygen distribution in a Hediste diversicolor burrow environment was investigated in a laboratory experiment using a 6-mm thick tank equipped with oxygen planar optodes. The two-dimensional oxygen distribution in a complete burrow was monitored every 2 min for 4 h. Oxygen concentrations fluctuated over a scale of minutes in the burrow lumen and wall (up to 2 mm) reflecting the balance between worm ventilation activity and oxygen consumption. The magnitude of the three surrounding micro-horizons (oxic, oscillating and anoxic) induced by the intermittent worm ventilation was spatially and temporally variable within the structure. Oxygen variations appeared to be controlled by distance from the sediment–water interface and the direction of water circulation. Moreover, there was an apparent ‘buffer effect’, induced by the proximity to the overlying water, which reduced the variations of lumen and wall oxygen in the upper part of the structure. These results highlight the heterogeneous distribution and dynamics of oxygen associated with H. diversicolor burrows and ventilation activity. They also highlight the necessity of integrating this complexity into the current burrow-base models in order to estimate the ecological importance of burrowing species in coastal ecosystems.     

Mitochondrial dysfunction in glaucoma: understanding genetic influences

Glaucoma is the leading cause of irreversible blindness worldwide. This review aims to provide a greater understanding of the complex genetic influences that may lead to mitochondrial dysfunction and increase susceptibility to retinal ganglion cell (RGC) loss in primary open angle glaucoma (POAG), and thus elucidate potentially important pathophysiological pathways amenable to therapeutic intervention. Emerging evidence from genome wide association and other genetic studies suggests that changes in the mitochondrial DNA (mtDNA) and in nuclear DNA genes that encode mitochondrial proteins may influence mitochondrial structure and function and, therefore, contribute to the pathogenesis of POAG. We propose that a variety of genes (OPA1, MFN1, MFN2, CYP1B1, PARL, SOD2, SRBD1, GST, NOS3, TNFa and TP53) may each confer a background susceptibility to POAG in different populations having one common link: mitochondrial dysfunction. The relationship between polymorphisms in these genes and increasing risk for POAG is presented and the limitations of the available current knowledge are discussed.