<Superscript>31</Superscript>P NMR study of polyphosphate levels during different growth phases of <Emphasis Type="Italic">Phycomyces blakesleeanus</Emphasis>

The changes in relative polyphosphate content, estimated as the intensity ratio of core polyphosphate signal and intracellular inorganic phosphate signal from ³¹P NMR spectra, during the growth of Phycomyces blakesleeanus are reported. The ratio increases from 16 h to 28 h of growth, the minimum occurs at 32 h, followed by sharp increase up to 36 h, and a steady decrease afterwards. The changes in the biomass during mycelium growth showed steady increases, with a stagnation period between 32 h and 36 h during which a pronounced increase in the intensity ratio of core polyphosphates to intracellular inorganic phosphate signal occurred. The reduction of growth temperature from 22°C to 18°C significantly decreased the rate and intensity of growth, but the pattern of polyphosphate changes remained unchanged. The changes of the intensity ratio of core polyphosphates to intracellular inorganic phosphate signal are linked to characteristic stages of sporangiophore development. Analysis of core polyphosphates, intracellular inorganic phosphate and β-ATP signal intensities suggest the role of polyphosphates as an energy and/or a phosphate reserves during Phycomyces development.     

Ecological conditions and the distribution of alpine juniper (<Emphasis Type="Italic">Juniperus communis</Emphasis> subsp. <Emphasis Type="Italic">alpina</Emphasis>) in the Hrubý Jeseník Mts

In the Hruby Jeseník Mts of the Czech Republic, research was carried out from 2001–2005 aimed at completing an up-to-date census of alpine juniper [Juniperus communis subsp. alpine (Smith) Čelakovsky] and an evaluation of the overall health status of the populations, and at investigating the impacts of the main environmental factors on the viability of this species. 13 sites were identified with 283 individuals in total, but the sites differed dramatically in the number of recorded individuals. Comparisons with historical literature sources show that the species has been in decline. The main reasons for this decline include: a lack of suitable sites for colonization connected with a lack of adequate disturbance factors, competition from shading trees, and the presumed high age of the juniper populations combined with zero generative reproduction. More than two thirds of the individuals showed slight damage to their assimilation system. A health status of bad or very bad was determined for 5 % of the alpine juniper individuals. These trees in the Hruby Jeseník Mts are also exposed to pressure from some herbivore insects and mammals. The populations are not yet in a literally critical state, but considering the range of impacts affecting them, it will be necessary to pay significant attention to their conservation strategies. Some recommendations for future management are suggested.     

Mechanisms of attenuation of abdominal sepsis induced acute lung injury by ascorbic acid

Bacterial infections of the lungs and abdomen are among the most common causes of sepsis. Abdominal peritonitis often results in acute lung injury (ALI). Recent reports demonstrate a potential benefit of parenteral vitamin C [ascorbic acid (AscA)] in the pathogenesis of sepsis. Therefore we examined the mechanisms of vitamin C supplementation in the setting of abdominal peritonitis-mediated ALI. We hypothesized that vitamin C supplementation would protect lungs by restoring alveolar epithelial barrier integrity and preventing sepsis-associated coagulopathy. Male C57BL/6 mice were intraperitoneally injected with a fecal stem solution to induce abdominal peritonitis (FIP) 30 min prior to receiving either AscA (200 mg/kg) or dehydroascorbic acid (200 mg/kg). Variables examined included survival, extent of ALI, pulmonary inflammatory markers (myeloperoxidase, chemokines), bronchoalveolar epithelial permeability, alveolar fluid clearance, epithelial ion channel, and pump expression (aquaporin 5, cystic fibrosis transmembrane conductance regulator, epithelial sodium channel, and Na(+)-K(+)-ATPase), tight junction protein expression (claudins, occludins, zona occludens), cytoskeletal rearrangements (F-actin polymerization), and coagulation parameters (thromboelastography, pro- and anticoagulants, fibrinolysis mediators) of septic blood. FIP-mediated ALI was characterized by compromised lung epithelial permeability, reduced alveolar fluid clearance, pulmonary inflammation and neutrophil sequestration, coagulation abnormalities, and increased mortality. Parenteral vitamin C infusion protected mice from the deleterious consequences of sepsis by multiple mechanisms, including attenuation of the proinflammatory response, enhancement of epithelial barrier function, increasing alveolar fluid clearance, and prevention of sepsis-associated coagulation abnormalities. Parenteral vitamin C may potentially have a role in the management of sepsis and ALI associated with sepsis.     

A spectrophotometric procedure for the determination of activity of restriction endonucleases

A simple procedure basically applicable for the quantitative determination of activity of all restriction endonucleases is described. Native DNA immobilized on cellulose is used as a substrate; after the treatment by restriction endonucleases this DNA is released to the solution. Changes of the optical density of the solution containing solubilized DNA permit quantitative determination of the restriction endonuclease activity.     

Species Recognition During Substrate-Borne Communication in <Emphasis Type="Italic">Nezara viridula</Emphasis> (L.) (Pentatomidae: Heteroptera)

The information code in the temporal and spectral characteristics of the substrate-borne communication signals produced by insects has been primarily studied in insects in the suborder Auchenorrhyncha. In the present study we investigated which of the female calling song (FCS) parameters in Nezara viridula (L.) (Heteroptera, Pentatomidae) are essential for recognition by conspecific males. In playback experiments we measured male vibrational responsiveness to FCS signals varying in the durations of pulse trains and inter-pulse train intervals, repetition times, duty cycles, and dominant frequencies, and determined the preference range for each specific parameter. Males were able to distinguish songs of different temporal and frequency parameters and responded best to values characteristic of the song of conspecific females. Signal recognition is achieved on the basis of two temporal filters tuned to the durations of the pulse train and inter-pulse train interval. Males responded best to the dominant frequency characteristic of conspecific songs, which are tuned to the resonant properties of the herbaceous plants used for intraspecific signal transmission during communication.     

Energy transfer in the major intrinsic light-harvesting complex from Amphidinium carterae

Carbonyl carotenoids are important constituents of the antenna complexes of marine organisms. These carotenoids possess an excited state with a charge-transfer character (intramolecular charge transfer state, ICT), but many details of the carotenoid to chlorophyll energy transfer mechanisms are as yet poorly understood. Here, we employ femtosecond transient absorption spectroscopy to study energy transfer pathways in the intrinsic light-harvesting complex (LHC) of dinoflagellates, which contains the carbonyl carotenoid peridinin. Carotenoid to chlorophyll energy transfer efficiency is about 90% in the 530-550 nm region, where the peridinin S2 state transfers energy with an efficiency of 25-50%. The rest proceeds via the S1/ICT channel, and the major S1/ICT-mediated energy transfer pathway utilizes the relaxed S1/ICT state and occurs with a time constant of 2.6 ps. Below 525 nm, the overall energy transfer efficiency drops because of light absorption by another carotenoid, diadinoxanthin, that contributes only marginally to energy transfer. Instead, its role is likely to be photoprotection. In addition to the peridinin-Chl-a energy transfer, it was shown that energy transfer also occurs between the two chlorophyll species in LHC, Chl-c2, and Chl-a. The time constant characterizing the Chl-c2 to Chl-a energy transfer is 1.4 ps. The results demonstrate that the properties of the S1/ICT state specific for carbonyl carotenoids is the key to ensure the effective harvesting of photons in the 500-600 nm region, which is of vital importance to underwater organisms.     

Comparison of aggregate stability within six soil profiles under conventional tillage using various laboratory tests

Soil structure stability was studied in every diagnostic horizons of six soil types (Haplic Chernozem, Greyic Phaeozem, two Haplic Luvisols, Haplic Cambisol, Dystric Cambisol) using different techniques investigating various destruction mechanisms of soil aggregates. Soil aggregate stability, assessed by the index of water stable aggregates (WSA), varied depending on the organic matter content, clay content and pH_KCl. The presence of clay and organic matter coatings and fillings, and presence of iron oxides in some soils increased stability of soil aggregates. On the other hand periodical tillage apparently decreased aggregate stability in the Ap horizons. Coefficients of aggregate vulnerability resulting from fast wetting (KV ₁) and slow wetting (KV ₂) tests showed similar trends of the soil aggregate stability as the WSA index, when studied for soils developed on the similar parent material. There was found close correlation between the WSA index and the KV ₁ value, which depended also on the organic matter content, clay content and pH_KCl. Less significant correlation was obtained between the WSA index and the KV ₂ value, which depended on the organic matter content and clay content. Coefficients of vulnerability resulting from the shaking after pre-wetting test (KV ₃) showed considerably different trends in comparison to the other tests due to the different factors affecting aggregate stability against the mechanical destruction. The KV ₃ value depended mostly on cation exchange capacity, pH_KCl and organic matter content.     

Comparison of different real-time PCR chemistries and their suitability for detection and quantification of genetically modified organisms

Background

The real-time polymerase chain reaction is currently the method of choice for quantifying nucleic acids in different DNA based quantification applications. It is widely used also for detecting and quantifying genetically modified components in food and feed, predominantly employing TaqMan^® and SYBR^® Green real-time PCR chemistries. In our study four alternative chemistries: Lux?, Plexor?, Cycling Probe Technology and LNA^® were extensively evaluated and compared using TaqMan^® chemistry as a reference system.

Results

Amplicons were designed on the maize invertase gene and the 5'-junction of inserted transgene and plant genomic DNA in MON 810 event. Real-time assays were subsequently compared for their efficiency in PCR amplification, limits of detection and quantification, repeatability and accuracy to test the performance of the assays. Additionally, the specificity of established assays was checked on various transgenic and non-transgenic plant species. The overall applicability of the designed assays was evaluated, adding practicability and costs issues to the performance characteristics.

Conclusion

Although none of the chemistries significantly outperformed the others, there are certain characteristics that suggest that LNA^® technology is an alternative to TaqMan^® when designing assays for quantitative analysis. Because LNA^® probes are much shorter they might be especially appropriate when high specificity is required and where the design of a common TaqMan^® probe is difficult or even impossible due to sequence characteristics. Plexor? on the other hand might be a method of choice for qualitative analysis when sensitivity, low cost and simplicity of use prevail.

     

Resurrection of <Emphasis Type="Italic">Cortinarius coalescens</Emphasis>: taxonomy,chemistry, and ecology

Cortinarius coalescens Kärcher & Seibt is a rare European species of the subgenus Phlegmacium, section Phlegmacioides, neglected in recent molecular studies. New primers (CortF and CortR) designed for species in the section Phlegmacioides allowed to obtain ITS rDNA sequence data from the holotype collection of C. coalescens; according to the results, this epithet has priority over C. crassorum Rob. Henry ex Rob. Henry, C. pardinus Reumaux, and C. parargutus Bidaud, Moënne-Locc. & Reumaux. Morphological and ecological observations on recent collections of C. coalescens from the Czech Republic in comparison with the co-occurring C. largus are discussed. Nomenclatural and taxonomic comments on C. tomentosus Rob. Henry, C. balteatotomentosus Rob. Henry, and C. subtomentosus Reumaux are also provided. So far, C. coalescens is known with certainty from Germany, France, and the Czech Republic, where it grows in deciduous forests on acid to neutral soils. Arsenic and its compounds were determined in C. coalescens and related species of the section Phlegmacioides: C. largus, C. pseudodaulnoyae, and C. variecolor. Total arsenic concentrations were in the range 3.6–30.2 mg kg^?1 (dry matter) and arsenobetaine was the major arsenic compound.