Covalent binding of polychlorinated biphenyls to proteins by reconstituted monooxygenase system containing cytochrome P-450

Incubation in the presence of NADPH and molecular oxygen of ¹⁴C-labeled polychlorinated biphenyls (PCBs) and two tetrachlorobiphenyl (TCB) isomers with a reconstituted system containing NADPH-cytochrome P-450 reductase and cytochrome P-450, both purified from liver microsomes of phenobarbital(PB)-pretreated rabbits, led to covalent binding of radioactive metabolites of PCBs and TCBs to the protein components of the system. A rabbit liver cytosol fraction added to the system provided more binding sites for the activated metabolites and thus increased the extent of binding markedly. The binding reaction depended absolutely on the reductase, cytochrome P-450 and NADPH, and required dilauroyl phosphatidylcholine and sodium cholate for maximal activity. A further stimulation of the binding was attained by including cytochrome b₅ in the reconstituted system. Four forms of cytochrome P-450, purified from liver microsomes of PB- and 3-methylcholanthrene(MC)-treated rabbits and rats, were used to reconstitute the PCB- and TCB-metabolizing systems, and it was found that PB-inducible forms of the cytochrome from both animals were more active than those inducible by MC in catalyzing the PCB- and TCB-binding reaction. Sodium dodecyl sulfate(SDS)-polyacrylamide gel electrophoresis indicated that, in the system containing the reductase, cytochrome P-450 and cytochrome b₅, PCB metabolites bound to the reductase and cytochrome P-450, but not to cytochrome b₅. In the presence of the liver cytosol fraction, the binding took place to many cytosolic proteins in addition to the reductase and cytochrome P-450.     

皖南、赣北奥陶纪笔石立体标本形成环境的初步研究*

皖南、赣东北和赣西北地区奥陶纪笔石地层发育良好,笔石化石丰富。宁国组和胡乐组均为笔石相地层,但笔石的保存特点并不相同。立体保存的黄铁矿化笔石标本主要见于宁国组,而胡乐组的笔石几乎均为薄膜标本。在比较宁国组和胡乐组在岩性、颜色、化石、矿物和元素等方面的特点后发现,两者有较明显的差异。这表明宁国组和胡乐组形成时的环境是不同的,前者为弱还原环境,后者为较强的还原环境,而在研究区内影响笔石体立体保存的主要因素为还原环境和较高的铁含量。在还原环境下,铁可呈Fe~(2+)存在,笔石体内含有硫,死亡后经降解作用可生成H_2S;H_2S和Fe~(2+)结合可使笔石体黄铁矿化,从而使笔石体硬化而呈立体保存下来。宁国组的铁含量明显高于胡乐组,这似可以解释宁国组产有较多笔石立体标本的原因。     

春季高寒山区牧草低温保护物质变化与其脱冻适应间关系研究

为了了解牧草低温保护物质与其抗冷冻的关系,在高寒山区测定了几种多年生牧草根中低温保护物质的含量和几种蔗糖酶活力的变化。结果表明,在4月土壤解冻期间,无芒雀麦、花雀麦、垂穗披碱草和草地早熟禾根中的蔗糖含量分别下降了74．1％、66．2％、45．2％和69．6％,果糖含量分别下降了72．0％、38．0％、68．3％和84．6％,葡萄糖含量分别下降了66．7％、62．5％、42．2％和65．2％,脯氨酸含量分别下降了83．0％、51．0％、43．3％和70．6％,可溶性蛋白质变化不大;蔗糖合成酶、蔗糖磷酸合成酶活性下降,蔗糖分解酶活性增加,蔗糖分解酶活性是蔗糖合成酶活性的22～35倍,是蔗糖磷酸合成酶活性的55～80倍。研究结果说明,春季牧草萌发时,根中已糖含量的下降是蔗糖分解酶活性增加的结果,根中脯氨酸作为氮的贮藏物质被用于根的呼吸和蛋白质合成。春季牧草根的抗寒能力随着根中低温保护物质的减少而下降,这也说明牧草的低温保护物质和牧草的抗寒性密切相关。     

Characterization of bacterial artificial chromosome transgenic mice expressing mCherry fluorescent protein substituted for the murine smooth muscle α‐actin gene

Smooth muscle α actin (SMA) is a cytoskeletal protein expressed by mesenchymal and smooth muscle cell types, including mural cells (vascular smooth muscle cells and pericytes). Using Bacterial Artificial Chromosome (BAC) recombineering technology, we generated transgenic reporter mice that express a membrane localized cherry red fluorescent protein (mCherry), driven by the full‐length SMA promoter and intronic sequences. We determined that the founders and F1 progeny of five independent lines contain 1–3 copies of the mCherry‐substituted BAC vector. Furthermore, we characterized the expression of SMA‐mCherry in relation to endogenous SMA in the embryo and in adult tissues, and found that the transgenic reporter in each line recapitulated endogenous SMA expression at all time points. We were also able to isolate SMA expressing cells from embryonic tissues using fluorescence‐activated cell sorting (FACS). We demonstrated that this marker can be combined with other vital fluorescent reporters and it can be used for live imaging of embryonic cardiodynamics. Therefore, these transgenic mice will be useful for isolating live SMA‐expressing cells via FACS and for studying the emergence, behavior, and regulation of SMA‐expressing cells, including vascular smooth muscle cells and pericytes throughout embryonic and postnatal development. genesis 48:457–463, 2010. © 2010 Wiley‐Liss, Inc.     

The influence of invertebrates on the breakdown ofPotamogeton pectinatus L. in a coastal marina (Zandvlei,South Africa)

The influence of invertebrates upon the decomposition ofPotamogeton pectinatus L. in a coastal Marina system was examined over 112 days using litter bags. Invertebrate inclusion bags (2 mm mesh, 5 mm holes) registered a dry mass loss of 1% d^–1, while exclusion litter bags (80 µm mesh) produced a 0.4% mass loss d^–1 (a 2.5 fold difference). Losses of ash and N from inclusion bags were greater than those from exclusion bags (p < 0.05). There was a three fold difference between the two treatments in the time taken for litter to breakdown to half the initial stock: T_1/2 for inclusion bags = 43 d, exclusion bags = 130 d. In both treatments, minerals showed an expected rapid loss, due to leaching, with a subsequent slow increase relative to the dry material remaining. A total of nine invertebrate taxa was recorded from inclusion bags, with a peak biomass of 64 mg g^–1 dry massPotamogeton bag^–1 reached at 64 days after immersion. Grazing amphipods,Melita zeylanica Stebbing andAustrochiltonia subtenuis (Barnard), numerically dominated the litter bag fauna, whileM. zeylanica and nymphs of the zygopteran predatorIschnura senegalensis (Rambur) formed most of the biomass. Scanning Electron Microscopy indicated heavy grazing of micro-organisms by invertebrates, with major qualitative differences occurring 112 days after immersion. Invertebrates significantly accelerated the rate of litter breakdown through their feeding activities, assisting fragmentation and thus contributing to plant losses and also by increasing the surface area for microbial colonisation and attack.     

似鳡属的系统地位及种的分化(鲤形目:鲤科)

似鳡属(Luciocyprinus Vaillant)是鲤科鱼类中属于鲃系(陈湘粦等,1984)的一个属,下咽齿3行,臀鳍无硬刺,具分枝鳍条5根。过去只记载一个种,外形很像鳡鱼,是由于摄食习性相同而导致的趋同现象(林人端,1981),故称似鳡,在广西俗称墨线鳡,云南俗称杆条鱼。分布于西江水系,包括云南中南部的湖泊,与鳡鱼分布于同一水系,但各占一定的水域,互不重叠。 关于似鳡的拉丁名称,惯用Fustis vivus Lin,是林书颜1932年定的新属新种,     

Regulatory Mechanisms of Reproductive Effort in Plants II. Plasticity in Reproductive Energy Allocation and Propagule Output of Glycine max Merr. (Leguminosae) Cultivated at Varying Densities and Nitrogen Levels

Abstract Plasticity in growth, reproductive energy allocation (RA), and reproductive output were studied in Glycine max Merr. Cv. Enrei (Leguminosae) grown under varying densities and soil nitrogen levels.
Marked plastic responses were detected in individual biomass, the patterns of resource allocation to total reproductive structures (RA) and also to propagules, reproductive outputs, and propagule weight under changing densities and soil nitrogen levels. Plants cultivated at higher densities exhibited proportionately lower individual biomass, lower RA, lower seed output, and smaller seed size in response to increasing density and decreasing soil nitrogen levels, although some deviations were observed, especially in the highest density plot with no fertilization. Differences due to different N-levels were not as great as those to changing density, which may in part be due to the fact that soybean has nitrogen-fixing bacteria in root tubercles, just as in any other Leguminosae. Fecundity was also maintained at the similar high rates of 80–97% in all plots examined, although slight but steady decreases were noted with increasing density. This resemblance in fecundity may be due to its strong inbreeding system.
Another important finding was that seed production under limited resource availability, notably lack of ample solar radiation due to strong interference at higher density plots, is exceedingly costly. This was most clearly exhibited by a sharp increase in relative energy partitioning to a single propagule in response to the increased density, the relative energy cost to a single propagule (RA) increasing from one to seven-fold. The results obtained in this study coincide well with the findings made in other plants, e.g., Helianthus annuus, Oryza sativa , and Coix ma-yuen , with the same experimental designs.     

PHOTOACCLIMATION IN THE TROPICAL CORALLINE ALGA HYDROLITHON ONKODES (RHODOPHYTA, CORALLINACEAE) FROM A FRENCH POLYNESIAN REEF

Pigment concentration, in vivo absorption, and photosynthetic parameters of the coralline alga Hydrolithon onkodes (Heydrich) Penrose and Woelkerling were compared among samples from a lagoon and from a reef crest of Tahiti Island. Four groups of specimens were considered, differing in their natural exposure to PAR. For specimens collected from the lagoon, the tissues from low-light samples had significantly higher pigment concentration, particularly chl a and phycobilins, compared with the high-light exposed plants that contained more total carotenoids. The in vivo absorption spectra normalized to chl a (called a* values) also revealed differences. The low-light samples had a reduced absorption capacity and a well-marked phycobilin absorption signature, whereas sunlit samples showed a greater absorption at wavelengths absorbed mainly by chl a and carotenoids. The decrease of a* when pigment concentration increased is interpreted as a consequence of the pigment packaging. Significantly lower α (chlorophyll basis) and higher E_k values were found in the shaded plants. The values of P _max for the four groups of specimens were not significantly different. The samples showed various degrees of photoinhibition depending on the light exposure during growth, and this effect was more pronounced in the shaded plants. The specimens from the reef crest deviate from the general model presented for the lagoon samples and show a mix of sun- and shade-exposed characteristics. We have shown that the coralline alga H. onkodes responds to its light environment, probably by acclimation rather than ecotypic genetic variation, by adjusting its physiology, but some morphological differences are also involved. Photoacclimation can explain partly the wide distribution of this species over the reef ecosystem and its major contribution to the building of the reef.     

Biological and physicochemical characterization of recombinant human erythropoietins fractionated by Mono Q column chromatography and their modification with sialytransferase

Ten erythropoietin (EPO) fractions differing in sialic acid content, ranging from 9.5 to 13.8 mol mol^–1 of EPO, were obtained from baby hamster kidney cell-derived recombinant human EPO by Mono Q column chromatography. The mean pI values of the EPO fractions determined by IEF-gel electrophoresis systematically shifted from 4.11 to 3.31, coinciding with the sialic acid content, without a change in the constitution of asialo N-linked oligosaccharides of each fraction. Although a linear relationship between thein vivo bioactivity and the sialic acid content of the fractionated, samples was observed until 12.1 mol mol^–1 of EPO, there was no further increase in their activity over 12.4 mol mol^–1 of EPO. On the other hand, an inverse relationship between thein vitro bioactivity and sialic acid content of EPO was observed. Also, we showed that thein vivo bioactivity of some fractions with low sialic acid contents was increased after treatment with 2,6-sialyltransferase, but thein vivo bioactivity of the other fractions with high sialic acid contents was either decreased or not affected.Abbreviations EPO erythropoietin - rHuEPO recombinant human erythropoietin - hCG human chorionic gonadotropin - BHK baby hamster kidney - CHO Chinese hamster ovary - NeuAc N-acetyl neuraminic acid - Gal galactose - HRCs hemolyser-resistant cells - WST-1 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium Na - IEF isoelectric focusing - pI isoelectric point