A simple rapid immunoassay for S-adenosylhomocysteine in plasma

The measurement of plasma S-adenosylhomocysteine is a more sensitive indicator of the risk for vascular disease than is plasma homocysteine. Because the level of S-adenosylhomocysteine is normally in the nanomolar range, it has been difficult to measure and necessitated the development of complex fluorometric and mass-spectrophotometric methods. We have now adapted an existing immunoassay used for the measurement of homocysteine to the measurement of S-adenosylhomocysteine in plasma. This assay is sensitive down to the level of less than 0.1 pmol, and there is no interference by S-adenosylmethionine. The assay is carried out in microplates, allows the measurement of 12 samples per plate and can easily be carried out in a 4-h period. The method is applicable to plasma samples having S-adenosylhomocysteine concentrations ranging from 10 to 150 nM without dilution. The mean value for 16 normal subjects by this method was 18.9±1.4 nM (S.E.M.), compared with 17.8±1.4 nM obtained by a previously described method using two high-performance liquid chromatography columns with fluorescence derivatization. Mean values for seven cirrhotic patients were 46.5±3.3 nM by this new method compared with 44.6±5.3 by the former method. The ease and speed of this method should allow the widespread measurement of this important metabolite in laboratories without access to sophisticated equipment.     

刺槐幼苗根茎过渡区初生维管组织的观察

对不同苗龄(1-12d)的刺槐(Robinia pseudoacacia L.)幼苗进行观察,比较了其初生维管组织的发育变化。结果表明:根中的初生维管组织借助下胚轴过渡到子叶中,而茎中的初生维管组织与下胚轴中的维管组织仅是简单的连接。这与Esau等认为幼苗的外部形态可以分为根-下胚轴-子叶和上胚轴苗两个系统的观点是一致的。本研究亦证实了可以将刺槐幼苗的外部形态描述为这两个系统,二者的维管组织只是简单的连接,不存在过渡。     

Climate and pH as determinants of vegetation succession in Central European man‐made habitats

Questions: (1) What are the most important abiotic environmental variables influencing succession in central European man‐made habitats? (2) How do these variables interact with one another and with variation in community properties? Location: Central, western and southern parts of the Czech Republic. Habitats included old fields, urban sites, spoil heaps after coal mining, sites at water reservoirs, extracted sand pit and peatland and reclaimed sites in areas deforested by air pollution. Methods: We investigated vegetation patterns on 15 succes‐sional seres, sampled by the same methods. Time of succession over which the data were available ranged from 12 to 76 years. The cover of vascular plant species (in %) was estimated in 5 m × 5 m plots. The relationships between vegetation characteristics (species composition, total cover, cover of woody species, species number and rate of dominant species turnover) and 13 abiotic site factors, including climatic and soil variables, were tested using CCA ordination and regression models. Results: Substratum pH, the only substratum characteristic, and climate were the environmental variables significantly affecting the vegetation patterns in the course of succession. The rate of succession, measured as the turnover of dominant species, was significantly more rapid in lowland than in mountain climates. On alkaline soils, species numbers in succession increased towards warmer climates. However, acid soils prevented any increase in species numbers, regardless of the climate. Surprisingly, forms of nitrogen and contents of C, P and cations did not exhibit any significant effect on the vegetation characteristics studied. Conclusions: Our approach, to compare a number of seres, can contribute not only to our understanding of succession, but also to help restoration projects to predict vegetation change because the crucial environmental variables, as identified by this study, are easy to measure.     

Vegetation shifts towards wetter site conditions on oceanic ombrotrophic bogs in southwestern Sweden

Question: Is ombrotrophic bog vegetation in an oceanic region of southwestern Sweden changing in the same direction over a five year period (1999 ‐ 2004) as northwest European bogs in the last 50 years, i.e. towards drier and more eutrophic vegetation? Location: The province of Halland, southwestern Sweden. Methods: Changes in species composition were monitored in 750 permanently marked plots in 25 ombrotrophic bogs from 1999 to 2004. Changes in species occurrences and richness were analysed and a multivariate statistical method (DCA) was used to analyse vegetation changes. Results: The species composition changed towards wetter rather than drier conditions, which is unlike the general pattern of vegetation change on bogs in northwestern Europe. Species typical of wetter site conditions including most Sphagnum species increased in abundance on the bogs until 2004. The total number of species per plot increased, mostly due to the increased species richness of Sphagnum species. Nitrogen‐demanding (eutrophic) species increased in occurrence. Conclusions: Ombrotrophic bog vegetation in an oceanic region in Sweden became wetter and was resilient to short‐term climatic shifts, after three years of below normal precipitation followed by several years with normal precipitation levels. Shifts towards more nitrogen demanding species were rapid in this region where the deposition levels have been high for several decades.     

Endothelial cell and macrophage regulation of vascular smooth muscle cell calcification modulated by cholestane-3beta, 5alpha, 6beta-triol

The cellular and molecular mechanisms that mediate vascular calcification remain poorly understood. In our previous study, oxysterol cholestane-3beta, 5alpha, 6beta-triol (Triol) was shown to promote vascular smooth muscle cells (VSMCs) calcification. In this study, by using direct coculture, non-contact transwell coculture, and culture with conditioned media, we investigated the roles of endothelial cells (ECs) and macrophages in the regulation of VSMCs calcification in the absence or presence of Triol. In vitro calcification was induced by incubation of VSMCs with beta-glycerophosphate. The results showed that ECs inhibited VSMCs calcification, as manifested by the reduction of calcium deposition in extracellular matrix. This effect of ECs on calcification was via the secreted soluble factors. Furthermore, the stimulation of ECs by Triol had no influence on ECs inhibition of calcification. On the other hand, macrophages promoted VSMCs calcification via the secreted soluble factors such as reactive oxygen species, which was further enhanced by Triol. Our results supported the roles for ECs and macrophages in vascular calcification, modulated by oxysterols in atherosclerotic plaque.     

Root and vascular tissue-specific expression of glycine-rich protein AtGRP9 and its interaction with AtCAD5, a cinnamyl alcohol dehydrogenase,in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis>

The vascular tissue of roots performs essential roles in the physical support and transport of water, nutrients, and signaling molecules in higher plants. The molecular mechanisms underlying the function of root vascular tissue are poorly understood. In this study, we analyzed the expression pattern of AtGRP9, a salt stress-responsive gene encoding a glycine-rich protein, and its interacting partner, in Arabidopsis thaliana. Analysis of GUS or GFP expression under the control of the AtGRP9 promoter showed that AtGRP9 was expressed in the vascular tissue of the root; subcellular localization analysis further demonstrated that AtGRP9 proteins were localized in the cell wall and in the cytoplasm. Yeast two-hybrid analysis revealed that AtGRP9 interacted with AtCAD5, a major cinnamyl alcohol dehydrogenase (CAD) involved in lignin biosynthesis, for which tissue-specific distribution was comparable with that of AtGRP9. These results suggest that AtGRP9 may be involved in lignin synthesis in response to salt stress as a result of its interaction with AtCAD5 in A. thaliana.     

Greenhouse gas emissions from a constructed wetland in southern Sweden

This paper investigates the greenhouse gas emissions from a Swedish wetland, constructed to decrease nutrient content in sewage treatment water. To evaluate the effect of the construction in terms of greenhouse gas emissions we carried out ecosystem-atmosphere flux measurements of CO₂, CH₄ and N₂O using a closed chamber technique. To evaluate the importance of vascular plant species composition to gas emissions we distributed the measurement plots over the three dominating plant species at the field site, i.e., Typha latifolia, Phragmites australis and Juncus effusus. The fluxes of CO₂ (total respiration), CH₄ and N₂O from vegetated plots ranged from 1.39 to 77.5 (g m⁻² day⁻¹), −377 to 1387 and −13.9 to 31.5 (mg m⁻² day⁻¹) for CO₂, CH₄ and N₂O, respectively. Presence of vascular plants lead as expected to significantly higher total respiration rates compared with un-vegetated control plots. Furthermore, we found that the emission rates of N₂O and CH₄ was affected by presence of vascular plants and tended to be species-specific. We assessed the integrated greenhouse warming effect of the emissions using a Global Warming Potential over a 100-year horizon (GWP₁₀₀) and it corresponded to 431 kg CO₂ equivalents m⁻² day⁻¹. Assuming a 7-month season with conditions similar to the study period this is equal to 90 tonnes of CO₂ equivalents annually. N₂O emissions were responsible for one third of the estimated total greenhouse forcing. Furthermore, we estimated that the emission from the forested bog that was the precursor land to Magle constructed wetland amounted to 18.6 tonnes of CO₂ equivalents annually. Hence, the constructed wetland has increased annual greenhouse gas emissions by 71.4 tonnes of CO₂ equivalents for the whole area. Our findings indicate that management processes in relation to wetland construction projects must consider the primary function of the wetland in decreasing eutrophication, in relation to other positive aspects on for instance plant and animal life and recreation as well as possible negative climatic aspects of increased emissions of CH₄ and N₂O.     

Taurine transporter is expressed in vascular smooth muscle cells

Summary. The regulation of vascular smooth muscle cells (VSMCs) function by taurine has been a subject of increasing interest and investigation, and taurine is taken up into cells through a specific transporter system, the taurine transporter (TAUT). In the present study, we examined the expression of TAUT in VSMCs and the kinetic parameters of the uptake process of TAUT in VSMCs. RT-PCR and western blot demonstrated that the mRNA and protein of TAUT was expressed in VSMCs in vitro. Immunohistochemistry using antibody for TAUT revealed the expression of this protein in rat thoracic aorta. The maximal [³H]taurine uptake rate in VSMCs was 37.75 ± 3.13 pmol/min per mg of protein, with a K _m value of 5.42 ± 0.81 μM. Thus, VSMCs are able to express a functional taurine transporter. The regulation and detailed function of taurine and TAUT in VSMCs remain unclear, but our findings suggest a functional role for them in VSMCs metabolism.     

Localisation of lymphatic vessels and vascular endothelial growth factors-C and -D in human and mouse skeletal muscle with immunohistochemistry

The present study was aimed to localise lymphatic vessels and their growth factors in human and mouse skeletal muscle with immunohistochemistry and specific antibodies (VEGFR-3, LYVE-1, VEGF-C and VEGF-D). The largest lymphatic vessels were found in perimysial connective tissue next to the arteries and veins, as has been shown earlier with electron microscopy. As a new finding, we also found small LYVE-1 positive vessels in the capillary bed between muscle fibres. These vessels were located next to CD31 positive blood capillaries and were of the same size, but fewer in number. In addition, we described the localisation of the two main lymphangiogenic growth factor proteins, vascular endothelial growth factor-C and -D. Both proteins were expressed in skeletal muscle at mRNA and protein levels. VEGF-D was located under the sarcolemma in some of the muscle fibres, in the endothelia of larger blood vessels and in fibroblasts. VEGF-C protein was localised to the nerves and muscle spindles, to fibroblasts and surrounding connective tissue, but was not found in muscle fibres or endothelial cells. Our results are the first to suggest the presence of lymphatic capillaries throughout the skeletal muscle, and to present the localisation of VEGF-C and -D in the muscles. Electronic Supplementary Material Supplementary material is available to authorised users in the online version of this article at .