碧桃朝鲜球坚蚧的出蛰若虫空间格局及应用

采用多种聚集度指标、Iwao法和Taylor幂函数法分析朝鲜球坚蚧出蛰若虫在碧桃上的空间格局。结果表明,出蛰若虫在碧桃上呈聚集分布,分布以个体群形式存在。碧桃不同段位上的聚集度指标和Iwao法均表明其为聚集分布,而Taylor幂函数法测定为上、中段均匀分布,下段聚集分布,综合分析其成因是各样地受害程度及垂直方向虫口密度差异显著所致。聚集均数λ值的测定结果表明,该虫的聚集原因主要由出蛰若虫自身的生物学习性和环境因素所引起。此外,利用空间格局参数确定了理论抽样数和序贯抽样模型。     

Some Observations on the Irradiance and Carbon Fixation in Grane Langsø

Photosynthetically active radiation (PAR) was registered daily in 0.25 m and 11.25 m in 3 years. Upwards irradiance of green light was 1 % of downward irradiance. Incubator ¹⁴C experiments showed that the phytoplankton enhanced its carbon uptake substantially with increasing concentrations of the carrier CO₂ in the ampoules. Severe carbon limitation of photosynthesis occurred in spring and summer 1961, where the real carbon fixation was only about 34 % of that calculated by the usual procedure. Utilization of light and DIC by the phytoplankton and its compensation depth was determined. Photoinhibition occurred down to 10–11 m, and net primary production was nearly always positive in any depth down to the lake bottom.     

Identification of two new denitrifying strains of Rhodobacter sphaeroides

Abstract Highly specific polyclonal and antibodies against either nitrate, nitrite or nitrous oxide reductases from a photosynthetic denitrifying bacterium Rhodobacter sphaeroides f. sp. denitrificans were used to show the presence of immunologically reactive proteins in strains that Pellerin and Gest had shown to grow in the dark with nitrate as a terminal acceptor [9]. Two strains of this bacterium, namely 81-3 and 2.4.3 synthesized the three denitrifying enzymes and were capable of denitrification. Strains 81-1 and 2.4.1 (neotype) both expressed nitrate reductase activities but nitrite reductase was not detected since these strains did not reduce nitrite. They also did not grow in the dark with nitrate as a terminal acceptor. Each of strains 81-1, 81-3, 2.4.1 and 2.4.3 contain four plasmids. R. sphaeroides f. sp. denitrificans , however, contains only one large 108 kb plasmid, which is distinctly different in size from those detected in the other strains. This indicates that the 108 kb plasmid is not necessarily specific for denitrification.     

DEMONSTRATION OF ALKALINE PHOSPHATASE PARTICIPATION IN THE MINERALIZATION OF OSTEOBLASTS BY ANTISENSE RNA APPROACH

MC3T3-E1 cells grown with ascorbic acid express sequentially osteoblastic marker proteins such as alkaline phosphatase (ALPase) and then form a mineralized extracellular matrix (ECM) as a consequence of osteoblastic differentiation. To explore the functional roles of ALPase in the process of osteoblastic maturation, an inducible expression vector for antisense ALPase RNA was constructed and stably transfected into MC3T3-E1 cells. The expression of antisense ALPase RNA in the differentiated MC3T3-E1 transfectants reduced markedly the ALPase activity, which resulted in a significant decrease in the deposition of minerals upon prolonged culture. These findings demonstrated directly that ALPase participated in the mineralizationof ECM.     

Enumerating phytoplankton with an upright compound microscope using a modified settling chamber

A modified settling chamber is described which permits the use of an upright compound microscope for phytoplankton enumerations. The chamber is composed of a 75 × 51 mm rectangular or 70 mm round-glass microslide base with a 130 m thick piece of sheet styrene attached to the upper surface. A circular cell is cut into the styrene material making a 26 mm diameter chamber which is approximately 130–150 m deep. Settling procedures follow Utermöhl's technique, with the use of a 0.13–0.15 mm thick coverslip (50 × 45 mm) to cover the chamber. The overall thickness of the settling slide is 1.13 mm which does not impact on the optical requirements for most objectives, including oil immersion objectives. The chamber encloses a total volume of 69–80 mm³. No statistical differences are observed in cell, filament or colony counts between the new and traditional chambers. Furthermore, count comparisons at different cell concentrations using the new chambers give consistent results. Thus, the resolution and availability of an upright microscope makes the use of the new settling chamber an attractive method for phytoplankton counting, especially in teaching situations.     

Inhibition of large-conductance calcium-activated potassium channel by 2-methoxyestradiol in cultured vascular endothelial (HUV-EC-C) cells

2-Methoxyestradiol, an endogenous metabolite of 17β-estradiol, is known to have antitumor and antiangiogenic actions. The effects of 2-methoxyestradiol on ionic currents were investigated in an endothelial cell line (HUV-EC-C) originally derived from human umbilical vein. In the whole-cell patch-clamp configuration, 2-methoxyestradiol (0.3–30 μm) reversibly suppressed the amplitude of K⁺ outward currents. The IC ₅₀ value of the 2-methoxyestradiol-induced decrease in outward current was 3 μm. Evans blue (30 μm) or niflumic acid (30 μm), but not diazoxide (30 μm), reversed the 2-methoxyestradiol-induced decrease in outward current. In the inside-out configuration, application of 2-methoxyestradiol (3 μm) to the bath did not modify the single-channel conductance of large-conductance Ca²⁺-activated K⁺ (BK_Ca) channels; however, it did suppress the channel activity. 2-Methoxyestradiol (3 μm) produced a shift in the activation curve of BK_Ca channels to more positive potentials. Kinetic studies showed that the 2-methoxyestradiol-induced inhibition of BK_Ca channels is primarily mediated by a decrease in the number of long-lived openings. 2-Methoxyestradiol-induced inhibition of the channel activity was potentiated by membrane stretch. In contrast, neither 17β-estradiol (10 μm) nor estriol (10 μm) affected BK_Ca channel activity, whereas 2-hydroxyestradiol (10 μm) slightly suppressed it. Under current-clamp condition, 2-methoxyestradiol (10 μm) caused membrane depolarization and Evans blue (30 μm) reversed 2-methoxyestradiol-induced depolarization. The present study provides evidence that 2-methoxyestradiol can suppress the activity of BK_Ca channels in endothelial cells. These effects of 2-methoxyestradiol on ionic currents may contribute to its effects on functional activity of endothelial cells. Received: 27 November 2000/Revised: 13 April 2001     

Relationship Between Amino Acid Properties and Protein Stability: Buried Mutations

In order to understand the mechanism of protein stability and to develop a simple method for predicting mutation-induced stability changes, we analyzed the relationship between stability changes caused by buried mutations and changes in 48 amino acid properties. As expected from the importance of hydrophobicity, properties reflecting hydrophobicity are strongly correlated with the stability of proteins. We found that subgroup classification based on secondary structure increased correlations significantly, and mutations within -strand segments correlated better than did those in -helical segments, which may result from stronger hydrophobicity of the -strands. Multiple regression analyses incorporating combinations of three properties from among all possible combinations of the 48 properties increased the correlation coefficient to 0.88 and by an average of 13% for all data sets. Analyzing the stability of tryptophan synthase mutants with Glu49 replaced by all other residues except Arg revealed that combining buriedness, solvent-accessible surface area for denatured protein, and unfolding Gibbs free energy change increased the correlation to 0.95. Consideration of sequence and structural information (neighboring residues in sequence and in space) did not significantly strengthen the correlations in buried mutations, suggesting that nonspecific interactions dominate in the interior of proteins.     

Altering Cellular Signaling Pathways Enhance Gene Silencing Activity of shRNA,shRNA.Ribozyme,and shRNA.Antisense in Neuroblastoma Cells

1. RNA interference (RNAi) is a multicomponent machinery that operates in a sequence-specific manner to repress the expression of genes in most eukaryotic cells.2. Here we wanted to investigate in a murine neuroblastoma cell line (NBP2) (a) if replacement of the loop of the short hairpin RNA (shRNA) with a hammerhead ribozyme (shRNA.RZ) or an antisense oligonucleotide (shRNA.AS) would affect the efficacy of gene suppression, and (b) if activation or inhibition of signaling pathways would enhance the efficacy of shRNA, shRNA.RZ, and shRNA.AS complex in gene silencing.3. We used U6-driven expression of these shRNAs to target either a short-lived green fluorescent protein (d2EGFP) or an endogenous cyclophilin A (CyP-A) gene in a d2EGFP expressing NBP2 cell line (NBP2-PN25).4. Activation of the cAMP signaling pathway or inhibition of phosphatidylinositol 3-kinase (PI3K) enhanced the efficacy of shRNA and shRNA.RZ complex in reducing the expression of d2EGFP shRNA.RZ complex was as efficacious as shRNA in reducing the expression of d2EGFP and CyP-A shRNA.AS complex showed a slightly lower efficacy than shRNA alone in decreasing d2EGFP expression. In contrast, the U6-driven hammerhead ribozyme targeted to d2EGFP showed no gene silencing activity.5. This report describes novel strategies of modifying shRNA and altering signaling pathways to affect siRNA-mediated gene silencing in a neuronal cell line.     

Does the ‘Old Bag’ Make a Good ‘Wind Bag’?: Comparison of Four Fabrics Commonly Used as Exclusion Bags in Studies of Pollination and Reproductive Biology

BACKGROUND AND AIMS: Fabrics used in pollination bags may exclude pollen carried by biotic vectors, but have varying degrees of permeability to wind-borne pollen. The permeability of bags to wind-borne pollen may have important consequences in studies of pollination and reproductive biology. The permeability of four fabrics commonly used in the construction of pollination bags was examined. METHODS: Deposition of wind-borne pollen on horizontally and vertically oriented microscope slides was assessed on slides enclosed in pollination bags, as well as on control slides. KEY RESULTS: It was found that the permeability of fabrics to wind-borne pollen, as measured by deposition on both horizontally and vertically oriented slides, decreased with pore size. However, deposition on horizontal slides was always greater than on vertical slides for a given fabric; this could manifest itself as differential success of pollination of flowers in bags-dependent on flower orientation. CONCLUSIONS: Obviously, bags with mesh size smaller than most pollen grains are impermeable to pollen. However, material for such bags is very expensive. In addition, it was also observed that bags with even moderately small pore size, such as pores (approx. 200 microm) in twisted fibre cotton muslin, offered highly significant barriers to passage of wind-borne pollen. Such bags are sufficiently effective in most large-sample-size reproductive biology studies.