Environmental and auxin regulation of wood formation involves members of the Aux/IAA gene family in hybrid aspen

Indole acetic acid (IAA/auxin) profoundly affects wood formation but the molecular mechanism of auxin action in this process remains poorly understood. We have cloned cDNAs for eight members of the Aux/IAA gene family from hybrid aspen (Populus tremula L. x Populus tremuloides Michx.) that encode potential mediators of the auxin signal transduction pathway. These genes designated as PttIAA1-PttIAA8 are auxin inducible but differ in their requirement of de novo protein synthesis for auxin induction. The auxin induction of the PttIAA genes is also developmentally controlled as evidenced by the loss of their auxin inducibility during leaf maturation. The PttIAA genes are differentially expressed in the cell types of a developmental gradient comprising the wood-forming tissues. Interestingly, the expression of the PttIAA genes is downregulated during transition of the active cambium into dormancy, a process in which meristematic cells of the cambium lose their sensitivity to auxin. Auxin-regulated developmental reprogramming of wood formation during the induction of tension wood is accompanied by changes in the expression of PttIAA genes. The distinct tissue-specific expression patterns of the auxin inducible PttIAA genes in the cambial region together with the change in expression during dormancy transition and tension wood formation suggest a role for these genes in mediating cambial responses to auxin and xylem development.     

Properties of photoreductions by photosystem II in isolated chloroplasts. III. The effect of uncouplers on phenylenediamine shuttles across the membrane in the presence of dibromothymoquinone

In the presence of the plastoquinone antagonist dibromothymoquinone the photoreduction of ferricyanide by isolated chloroplast membranes is attributed to Photosystem II. The reaction is stimulated by the addition of phenylenediamine or C-substituted phenylenediamines (which may form a diimine on oxidation) but not of N-substituted phenylenediamines (which form a stable radical on oxidation). Phenylenediamines also restore NADP reduction (and O₂ evolution) in 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB)-treated chloroplasts. In this bypassing of the inhibition site, N-substituted phenylenediamines are very effective, whereas p-phenylenediamine and C-substituted phenylenediamines are inefficient. Uncouplers exhibit a surprising effect on these systems. Even under coupling conditions uncouplers inhibit electron flow to ferricyanide mediated by phenylenediamine in the pH range 7.3–8.0, whereas the restoration of the NADP system is stimulated.

For the interpretation of the results the side of the membrane involved is considered. It is proposed that in ferricyanide reduction by Photosystem II, a phenylenediimine/diamine shuttle operates which moves reducing equivalents from the inside to the outside across the membrane. This shuttle requires a pH gradient across the membrane because of different optimal ratios of diimine/diamine inside and outside. This pH difference is abolished by the uncoupler, accounting for the observed inhibition.

The restoration of electron flow from water to NADP in DBMIB-treated chloroplasts is assumed to be a bypass of the inhibition site inside the membrane via a phenylenediamine. Because the imine/amine ratio brought about by the pH gradient is not favorable for the inside oxidation an uncoupler stimulates NADP reduction even under coupling conditions.

Also in photoreductions by Photosystem I, for example NADP reduction at the expense of P-phenylenediamine/ascorbate, a shuttle of reducing equivalents across the membrane occurs but this time from outside to inside.     

Stomatal closure by ultraviolet radiation

The effect of ultraviolet radiation (UV) (255–325 nm) on stomatal closure was investigated on tef [ Eragrostis tef (Zucc) Trotter] in the presence of white light (ca 50 ·mol m⁻² s⁻¹). The action spectrum showed that UV (ca 2 ·mol m⁻² s⁻¹, half band width about 10 nm) of 285 nm or shorter wavelengths was very efficient in causing stomatal closure. The effectiveness decreased sharply towards longer wavelengths. Radiation of 313 nm or longer wavelengths was practically without effect. Increasing UV intensity increased stomatal resistance. When stronger white light (5 to 9 times stronger than the one used during irradiation) was administered, stomates re-opened rapidly irrespective of whether the UV was on or off, although a subsequent gradual closing tendency was observed when the UV was on.     

A new species of Otostegia (Lamiaceae) from Somalia

A new species from gypsum plains in northern Somalia, Otostegia ericoidea Ryding is described and illustrated.     

Reproduction of Zoanthus sansibaricus in the infra-littoral zone at Taisho Lava Field, Sakurajima, Kagoshima, Japan

In order to obtain data on the reproductive pattern of the clonal cnidarian Zoanthus sansibaricus, polyps were sampled by scuba gear at Taisho Lava Field, Sakurajima, for 24 months between April 2000 and March 2002 (polyps collected weekly for breeding season). According to cross-sections, Zoanthus polyps were divided into three sexual types; male, female and asexual, and were found in the same colony. At Sakurajima, Zoanthus sansibaricus spawned in the middle of July, releasing oocytes and sperm. These spawning events occur synchronously with moon phase. In gametogenesis of Zoanthus sansibaricus, oocytes became recognizable in February and grew rapidly from the end of June onward. Spermatocytes became recognizable in June and matured rapidly in the middle of July. After spawning events, oocytes still remaining in the endoderm were absorbed into Zoanthus tissue quickly.     

Fluorescent detection of beta-lactamase activity in living Escherichia coli cells via esterase supplementation

The TEM-1 beta-lactamase protein fragment complementation assay was investigated for its applicability in affinity protein-based interaction studies in Escherichia coli, using an affibody-based model system. Results from co-transformation experiments showed that an ampicillin resistant phenotype was specifically associated with cognate affibody-target pairings. Attempts to monitor beta-lactamase complementation in vitro with the fluorescent beta-lactamase substrates CCF2/AM and CCF2 showed that E. coli lacks an esterase activity necessary for activation of the esterified and membrane-permeable CCF2/AM form of the substrate. Interestingly, supplementation of the assay reaction with a purified fungal lipase (cutinase) resulted in efficient activation of CCF2/AM in vitro. Further, periplasmic expression of cutinase allowed for fluorescent discrimination between beta-lactamase positive and negative living E. coli cells using the CCF2/AM substrate, which should open the way for novel applications for this prokaryotic host in protein interaction studies.     

Regulation of 5-lipoxygenase enzyme activity

In this article, regulation of human 5-lipoxygenase enzyme activity is reviewed. First, structural properties and enzyme activities are described. This is followed by the activating factors: Ca2+, membranes, ATP, and lipid hydroperoxide. Also, studies on phosphorylation of 5-lipoxygenase and nuclear localization sequences are reviewed.     

Morning versus evening dosing of desloratadine in seasonal allergic rhinitis: a randomized controlled study [ISRCTN23032971].

BACKGROUND: A circadian rhythm of symptoms has been reported in allergic rhinitis and some studies have shown the dosing time of antihistamines to be of importance for optimizing symptom relief in this disease. The objective of this study was to examine the efficacy of morning vs. evening dosing of the antihistamine desloratadine at different time points during the day. METHODS: Patients >/= 18 years, with seasonal allergic rhinitis received desloratadine 5 mg orally once daily in the morning (AM-group) or evening (PM-group) for two weeks. Rhinorrhea, nasal congestion, sneezing and eye symptoms were scored morning and evening. Wilcoxon rank sum and 2-way ANOVA test were used. RESULTS: Six-hundred and sixty-three patients were randomized; 336 in the AM-group; 327 in the PM-group. No statistically significant differences were seen between the AM and PM group at any time points. In the sub-groups with higher morning or evening total symptom score no difference in treatment efficacy was seen whether the dose was taken 12 or 24 hours before the higher score time. There was a circadian variation in baseline total symptom score; highest during daytime and lowest at night. The circadian variation in symptoms was reduced during treatment. This reduction was highest for daytime symptoms. CONCLUSIONS: A circadian rhythm was seen for most symptoms being more pronounced during daytime. This was less apparent after treatment with desloratadine. No statistically significant difference in efficacy was seen whether desloratadine was given in the morning or in the evening. This gives the patients more flexibility in choosing dosing time.     

Fluorescence-microscopy-based image analysis for analyte-dependent particle doublet detection in a single-step immunoagglutination assay

A novel fluorescence-microscopy-based image analysis method for classification of singlet and doublet latex particles is demonstrated and applied to a particle-based immunoagglutination assay for quantification of biomolecules in microliter-volume bulk samples. The image analysis method, verified by flow cytometric agglutination analysis, is based on a pattern recognition algorithm employing Gaussian-base-function fitting which allows robust identification and counting of singlets, doublets, and higher agglomerates of fluorescent microparticles. The immunoagglutination assay is experimentally modeled by a biotin-streptavidin interaction, with the goal of both theoretically and experimentally investigating the performance of a general immunoagglutination-based assay. For this purpose a theoretical model of the initial agglutination kinetics, based on particle diffusion combined with a steric factor determined by the level of specific and nonspecific agglutination, was developed. The theoretical model combined with the experimental data can be used to optimize an agglutination-based assay with regard to sensitivity and dynamic range and to estimate the affinity, receptor surface density, molecular and binding site sizes, and level of nonspecific binding that is present in the assay. The experimental results are in good agreement with the theoretical model, indicating the usefulness of the model for immunoagglutination assay optimization.     

Synthesis of a disaccharide fragment of rhamnogalacturonan II

A disaccharide portion of the A-side chain of the rhamnogalacturonan II oligosaccharide has been prepared. Glycosylation of methyl (methyl 3,4-O-isopropylidene-alpha-D-galactopyranosid)uronate with p-tolyl 2,3-di-O-acetyl-3-C-(benzyloxymethyl)-1-thio-alpha/beta-D-erythrofuranoside was carried out using N-iodosuccinimide as promoter and silver trifluoromethanesulfonate as catalyst. Removal of the protecting groups gave the beta-d-Apif-(1-->2)-alpha-D-GalpA-OMe disaccharide.