Influence of hydrogen fluoride fumigation on the water economy of soybean plants

The influence of hydrogen fluoride fumigation on water economy was studied using soybean (Glycine max [L] Merr.). Fumigation caused partial stomatal closure in 1 hour and practically complete closure within 4 hours. The transpiration rate was greatly reduced by fumigation, while the leaf temperature was increased. Water potential increased after 1 day but fell drastically when necrosis occurred. Effects of interrupted fumigation during the day were somewhat less severe in all respects than those of continuous fumigation; nighttime fumigation caused only minor effects. Fluoride uptake was also much less from nighttime than from daytime fumigations.     

125I]iodohydroxynitrobenzylthioinosine: a new high-affinity nucleoside transporter probe

[125I]iodohydroxynitrobenzylthioinosine ([125I]IH-NBMPR), a new gamma-labeled nucleoside transport inhibitor, has been prepared at a theoretical specific activity of 2000 Ci/mmol (1 Ci = 37 GBq). IH-NBMPR was more acidic than hydroxynitrobenzylthioinosine (H-NBMPR), having a pKa of 4.6. Site-specific binding of [125I]IH-NBMPR to membrane-enriched fractions (MEF) from S49 mouse lymphoma cells was pH dependent, increasing with the fraction of undissociated molecules present; it was maximal at pH 4.5 and negligible at pH 7.0. Scatchard analysis of specific binding to MEF from S49 cells under equilibrium conditions at pH 5.0 yielded a Kd of 15 nM (equivalent to 4.0 nM for the undissociated fraction of inhibitor molecules) and maximum number of binding sites (Bmax) of 4.9 pmol/mg protein. Specific binding of IH-NBMPR could not be demonstrated in MEF from AE1 cells, a nucleoside transport-deficient mutant of S49 cells. Influx of uridine into mouse erythrocytes at pH 5.0 in the presence of 5 microM IH-NBMPR (1.4 microM undissociated IH-NBMPR) was reduced to about 7% of the control value, indicating that this compound is an effective nucleoside transport inhibitor. Photoactivation of site-bound [125I]IH-NBMPR, following equilibration of the ligand with MEF from S49 cells at pH 5.0, resulted in specific covalent labeling of a polypeptide with a relative molecular mass of 52,000-63,000, identified on sodium dodecyl sulfate-polyacrylamide gels. These results indicate that the new, iodinated ligand is an inhibitor of nucleoside transport and that it binds specifically and with high affinity to nucleoside transporter polypeptides in mammalian cells.     

Species detection and delineation in the marine planktonic diatoms Chaetoceros and Bacteriastrum through metabarcoding: making biological sense of haplotype diversity

High-throughput sequencing (HTS) metabarcoding is commonly applied to assess phytoplankton diversity. Usually, haplotypes are grouped into operational taxonomic units (OTUs) through clustering, whereby the resulting number of OTUs depends on chosen similarity thresholds. We applied, instead, a phylogenetic approach to infer taxa among 18S rDNA V4-metabarcode haplotypes gathered from 48 time-series samples using the marine planktonic diatoms Chaetoceros and Bacteriastrum as test case. The 73 recovered taxa comprised both solitary haplotypes and polytomies, the latter composed each of a highly abundant, dominant haplotype and one to several minor, peripheral haplotypes. The solitary and dominant haplotypes usually matched reference sequences, enabling species assignation of taxa. We hypothesise that the super-abundance of reads in dominant haplotypes results from the homogenization effect of concerted evolution. Reads of populous peripheral haplotypes and dominant haplotypes show comparable distribution patterns over the sample dates, suggesting that they are part of the same population. Many taxa revealed marked seasonality, with closely related ones generally showing distinct periodicity, whereas others occur year-round. Phylogenies inferred from metabarcode haplotypes enable delineation of biologically meaningful taxa, whereas OTUs resulting from clustering algorithms often deviate markedly from such taxa.     

Siderophore production by Fusarium venenatum A3/5

Fusarium venenatum A3/5 was grown in iron-restricted batch cultures and iron-limited chemostat cultures to determine how environmental conditions affected siderophore production. The specific growth rate in iron-restricted batch cultures was 0.22 h(-1), which was reduced to 0.12 h(-1) when no iron was added to the culture. D(crit) in iron-limited chemostat culture was 0.1 h(-1). Siderophore production was correlated with specific growth rate, with the highest siderophore production occurring at D=0.08 h(-1) and the lowest at D=0.03 h(-1). Siderophore production was greatest at pH 4.7 and was significantly reduced at pHs above 6.0. Siderophore production could be enhanced by providing insoluble iron instead of soluble iron in continuous flow cultures.     

Engagement of ubiquitination and de-ubiquitination at rostral ventrolateral medulla in experimental brain death

Background

Whereas brain death is a vitally important clinical phenomenon, our contemporary understanding on its underlying cellular mechanisms remains elusive. This study evaluated whether the ubiquitin-proteasome system (UPS) in the rostral ventrolateral medulla (RVLM), a neural substrate that our laboratory identified previously to be intimately related to brain death, is engaged in this fatal process.

Methods

We performed proteomics, Western Blot, real-time PCR, ELISA and pharmacological experiments in conjunction with a clinically relevant experimental endotoxemia model of brain death based on intravenous administration of Escherichia coli lipopolysaccharide in adult male Sprague–Dawley rats.

Results

Proteomics, Western blot and enzyme activity analyses demonstrated that polyubiquitination was preserved and de-ubiquitination by ubiquitin C-terminal hydrolase isozyme-L1 (UCH-L1) was sustained, alongside increased monoubiquitin availability or proteasome activity in RVLM over the course of experimental endotoxemia. However, real-time PCR revealed no significant alteration in proteasome subunit alpha type-1, ubiquitin or UCH-L1 at mRNA level. Functionally, whereas microinjection into the bilateral RVLM of proteasome inhibitors (lactacystin or proteasome inhibitor II) potentiated survival, an inhibitor of ubiquitin-recycling (ubiquitin aldehyde) or an UCH-L1 inhibitor exacerbated mortality.

Conclusions

We proposed previously that the progression towards brain death entails a tug-of-war between pro-death and pro-life programs in RVLM. It is conceivable that ubiquitination or de-ubiquitination in RVLM participate in brain death by regulating the degradation of the proteins involved in those programs.     

A one-pot synthesis of 1-alpha- and 1-beta-D-arabinofuranosyl-2-nitroimidazoles: synthons to the markers of tumor hypoxia

1-alpha- and 1-beta-D-Arabinofuranosyl-2-nitroimidazole (alpha-AZA and beta-AZ A) are synthons for a number of potential markers of tissue hypoxia. A one pot synthesis in which 2-nitroimidazole is coupled with a mixture of alpha- and beta-1-O-acetyl-2,3,5-tri-O-benzoyl-D-arabinofuranose in the presence of stannic chloride, followed by deprotection using ammonia/methanol, is described Previously reported conditions for coupling 2-nitroimidazole to 1-alpha-bromoarabinofuranose protected by base-hydrolyzable groups afforded alpha-AZA almost exclusively.     

Matric potential of several plant tissues and biocolloids

The pressure membrane apparatus was used to study the matric potential (imbibition pressure or moisture tension) of plant tissues and of several organic colloidal preparations.