Microscopic insight into the DNA condensation process of a zwitterion‐functionalized polycation

Zwitterion‐functionalized polycations are ideal gene carriers with long circulation, high cellular uptaking and low cell viability. However, the trade‐off between the DNA condensation efficiency and the cell viability must be addressed. The purpose of this study is to provide a microscopic insight into the DNA condensation process and to explore the effect of a zwitterionic block of zwitterion‐functionalized polycation, which is of great significance in designing novel gene delivery systems. Poly[2‐(dimethylamino)ethyl methacrylate‐b‐(sulfobetaine methacrylate)] (PDMAEMA‐b‐PSBMA) copolymers were synthesized and used as the model systems. Different from the conventional concept that the PSBMA zwitterionic block act only as the “stealthy” groups, the subtle differences in physical and colloidal characteristics between the polycation/DNA polyplexes show that the PSBMA segment is capable of wrapping DNA attributed to the quaternary ammonium cations, without compromising the DNA condensation capability. On the other hand, the incorporation of PSBMA block reduces the surface charge of the polyplexes, which substantially result in the inefficient transfection and the reduced cytotoxicity.     

不同耕作方式下草甸栗钙土燕麦田土壤微生物特征

河北坝上地区高寒半干旱的独特生态环境,土地沙化风蚀严重,作物产量低,土壤微生物的活动尚未有深入研究,尤其是人为干扰下的土壤微生物在作物生长季的动态变化。为了明确耕作方式对草甸栗钙土土壤微生物性状的影响特征,依托定位12a的免耕、深松、常规耕作田间试验基础,通过辅助设置12a免耕、深松后的翻耕处理,监测了燕麦田土壤微生物量碳、活跃微生物量和土壤呼吸速率等性状。结果表明,土壤微生物量碳与活跃微生物量围绕燕麦抽穗期为"W型"时序变化,长期免耕与深松下呈现0—10 cm表层土壤富集微生物量碳的空间分布特征。免耕与深松有利于提高0—10 cm土层土壤微生物量,多年免耕和深松后翻耕能使土层间土壤活跃微生物量差异减小。燕麦田土壤呼吸速率呈现"U型"时序变化,免耕0—10 cm土层呼吸速率具有较其他耕作方式更高的趋势。在燕麦生育期内,土壤呼吸商一直处于较低而平稳的水平,燕麦收获后进入土壤微生物的高活性期;0—10 cm土层翻耕与多年免耕与深松后的翻耕处理土壤呼吸商有高于免耕处理的趋势。翻耕对于促进冷凉区土壤库存养分的活化与适时供应具有重要作用。     

Microspectrophotometric measurements of DNA by automated computerized scanning in cycling cells of theChrysanthemum segetum shoot apex

Summary A new technique of exploitation of the data was proposed after DNA scanning microdensitometry. By using all of the measurements obtained from the seriated sections of a single nucleus, this method made it possible to estimate six characteristic parameters during the different phases of the cell cycle in the various shoot apical cells. The cells whose rate of proliferation was the highest showed the biggest variations of their nuclear and nucleolar volumes during the cell cycle. In the axial zone, where the cells have a slow cell cycle and display the longest duration of the G₁ phase, the volume occupied by dispersed DNA was greater than in the cells of the lateral zone and of the rib meristem, where the cell cycle and the G₁ phase were short. No matter what the cell type, the proportion of the dispersed and condensed DNA varied little when the G₁ and G₂ phases were compared. In the Z phase, characterized by a decondensation of the DNA, the mean DNA amount was 3.4 C. The evolution of the nuclear density during the interphase was also estimated. It is demonstrated that the main feature of the shoot apex zonation was the decondensation of the condensed DNA in the axial zone in both the G₁ and G₂ phases.     

Nuclear morphogenesis during spermiogenesis in the nudibranch molluscHypselodoris tricolor (Gastropoda,opisthobranchia)

An electron microscope study was carried out on Hypselodoris tricolor spermatids to describe the development of the nuclear morphogenesis and investigate the possible cause(s) of the change in the shape of the spermatid nucleus during spermiogenesis. Three different stages may be distinguished in the course of the nuclear morphogenesis on the basis of the morphology and inner organization of the nucleus. Stage 1 spermatid nuclei are spherical or ovoid in shape and the nucleoplasm finely granular in appearance. Stage 2 nuclei exhibit a disc- or cup-shaped morphology, and the chromatin forms short, thin filaments. During stage 3, a progressive nuclear elongation takes place, accompanied by chromatin rearrangement, first into fibers and then into lamellae, both formations helically oriented. A row of microtubules attached to the nuclear envelope completely surrounds the nucleus. Interestingly, the microtubules always lie parallel to the chromatin fibers adjacent to them. Late stage 3 spermatids show the highest degree of chromatin condensation and lack the manchette at the end of spermiogenesis. Our findings indicate the existence of a clear influence exerted on the chromatin by the manchette microtubules, which appear to be involved in determining the specific pattern of chromatin condensation in Hypselodoris tricolor.     

Recording mitotic chromosome cycle induction in living plant cells

Summary The presence of a prophase nucleus inHaemanthus endosperm happens to trigger the break down of the nuclear envelope in any interphase nucleus, located in its close proximity. Besides, chromosomes in the interphase nucleus start condensing gradually for the initial breaking point which is the nearest point to the prophase. The observation suggest the diffusion of an inducer, whose progression has been recorded to occur at a rate of 1.1 m/min.     

Association of protein amount polymorphism (PAP) among maize lines with performances of their hybrids

Summary It has been suggested that molecular foundations of phenotypic diversity reside in the variability of genome expression. This variability can be appraised through the polymorphism of individual protein amounts (PAP: protein amount polymorphism). Eight maize inbred lines and ten of their single-cross hybrids were analyzed by two-dimensional polyacrylamide gel electrophoresis in order to examine the potential of PAP for predicting hybrid vigor. The 28 possible pairs of lines were characterized for: (i) the number, H of expected heterozygous structural loci in their hybrid, in the sample of loci revealed by 2D-PAGE; (ii) four distance indices based on PAP; (iii) the hybrid values for five agromorphological characters measured in four different year/locations. For the subset of ten hybrids analyzed by 2D-PAGE, the number of cases of nonadditive inheritance (NA) was also counted. Whereas H appeared to be related neither to the PAP indices, nor to NA, nor to hybrid performances, PAP indices were correlated to NA, and both were positively associated to hybrid performances. The possibility that PAP is responsible for quantitative trait variation is discussed. This could result in the definition of biological predictors of heterosis.     

Phosphorylation state of protamines 1 and 2 in human spermatids and spermatozoa

The basic nuclear proteins of a fraction of elongating spermatids from human tests and of a fraction of motile spermatozoa from the ejaculate, separated by ion-exchange chromatography, were compared. Analysis by acetic acid-urea polyacrylamide gel electrophoresis (PAGE) showed that, in both fractions, four proteins of lower mobility were coeluted with protamine 1 by 23% guanidinium chloride (GuCI) while protamine 2 alone was eluted by 50% GuCI. Treatment with alkaline phosphatase identified those four proteins as phosphorylated protamines, and cyanogen bromide (CNBr) treatment of the dephosphorylated protamines distinguished them as variants of protamine 2 and not of protamine 1. Thus far, phosphorylated forms of protamine 1 have not been detected in either spermatids or spermatozoa. Those observations indicate that protamine 2 functions in the cycle of phosphorylation-dephosphorylation, which is essential to the process of sperm chromatin condensation, while the role of protamine 1 in human spermiogenesis is not yet defined. The presence of phosphorylated protamine in motile, presumably mature spermatozoa appears to be characteristic of human sperm but not of the sperm of other mammals and is probably the basis for the heterogeneity of chromatin condensation frequently observed in human spermatozoa.     

A minireview of microheterogeneity in H1 histone and its possible significance

Subtypes of H1 histone vary in primary structure, and the higher organisms that have been studied each seem to have about a half-dozen subtypes. The proportions of these subtypes vary with the progress of differentiation as seen in embryonic development, hormonally induced changes, spermatogenesis, and terminal differentiation. The H1 subtypes differ among themselves in their ability to condense DNA and small chromatin fragments. They have the potential, therefore, of causing different parts of the chromatin to be condensed to different degrees.