白刺珠心组织及其营养功能的超微结构观察

白刺具厚珠心,珠心组织发达。原胚期,根据超微结构特征,珠心组织大体可区分为三层:外围不活跃细胞层、特化细胞层和胚囊外围解体细胞层。珠心细胞解体时,细胞壁首先以微纤丝状态分散,原生质不同程度破坏,并累积脂滴。近胚囊壁降解后,珠心细胞即成为开放细胞,原生质团游离到胚囊周围。胚囊壁在珠孔区和合点区间断,因之,珠心细胞的游离原生质团可直接迁入胚囊内。在胚囊壁和中央细胞质膜之间存在明显间隙区域,其中有大量壁旁体存在,是珠心细胞降解物质进入胚囊的另一重要途径。     

高等植物衰老叶片中原生质的撤退现象以及原生质运动在有机物运输中可能具有的作用

高等植物体内有机物的运输问题,通常研究的主要是富含糖分的汁液在筛管中的大量流动。衰老叶片中细胞内含物的彻底转移,由于涉及不少含多种元素的复杂有机物,也应是有机物运输中值得考虑的一个重要方面。这不仅是因为它在理论上与运输的其它方面有密切牵连,而且在生产上也有它的现实意义。行将收获籽粒的小麦、玉米。其茎叶在衰退过程中细胞内含物能否彻底地转移,会显著地影响籽粒的产量与品质。本文在葱属、小麦等衰老叶鞘中,以核物质的变动为主要指标,按照原生质解体与撤退的顺序作了系统的观察。试验结果充实了我们过去提出的建议:衰老叶片中细胞内含物的彻底转移,是靠局部解体的原生质自身的胞间运动,原生质于是大量汇集到叶脉的维管束,再从中向外转移,由于来不及撤离出去,它时常入侵导管。原生质的局部解体最明显的表现就是胞核的解体与核物质的释放及其向维管束转移过程中状况的逐渐改变。植物枯竭叶片中时常出现的大量钙盐结晶就是原生质解体时遗留下来的后果。不仅衰老组织中物质的转移是靠原生质的胞间运动,过去的工作表明,就是急剧生长的幼嫩组织也出现原生质的胞间运动。高等植物原生质的胞间运动可以看作是细胞运动因有胞壁阻隔而表现的形式。正是象生物界广泛出现的造形运动一样,高等植物细胞原生质只有从原属的衰老器官靠自己的运动才能转移到新生部位,而淀粉等储藏物质则必需降解到糖才能随汁液运输。因此,我们提出建议:筛管运输既有迅速的汁液集体流动,也有缓慢的原生质移动。根据已有的证据。许多生物的各种原生质运动,不论是它全身的运动,还是内部透明质的流动,都可能是具有ATP酶活性的收缩蛋白来推动的。因此,这两种筛管运输也可能是靠收缩蛋白在与原生质结构的不同结合下进行。叶片由大量汁液输出为主的运输完全转到原生质自身的撤退,关键就在于它衰老到不能恢复的状态,这可能和衰老叶片胞核解体的出现联系在一起。     

渗透稳定剂的互补效应及某些渗透稳定剂对蓝藻细胞壁的降解作用

由多种盐组成的复合渗透稳定剂用于分离蓝藻原生质球的效力与单一盐溶液相比较,其作用多数显示加强,但对原生质球稳定性的影响随盐类组合而异。若干种盐类,如酒石酸铵、硫酸铵和硫酸镁,对蓝藻细胞壁表现一定的降解作用,以酒石酸铵作用最强,可用于分离原生质球。此种原生质球透明度较差,但对低渗敏感     

紫露草雄蕊毛细胞中质膜体超微结构的观察

质膜体是由质膜的内折或外突而形成的。Harris等”’把质膜的外突称为原生质小管（Plasmatubules）,认为原生质小管是一种特化的质膜体。质膜体或原生质小管为质膜的一种异型体结构,明显增加质膜的表面积。由于植物细胞存在着细胞间短距离的物质运输,增加膜的表面积可扩大物     

分离蓝藻原生质球的新方法：青霉素－溶菌酶法

蓝藻原生质球(体)的研究,至今尚未取得决定性突破,成为细胞壁生物之中的唯一例外。这不仅表现在原生质球(体)的再生没有成功,而且分离制备方法亦存在很大问题。从1967年沿用至今的Crespi-Biggins溶菌酶法不仅在质量上不能得到真正理想的原生质球,而且数量上也达不到。由于在分离过程中细胞大量破裂,原生质球的产率很     

染色体DNA转化原生质球构建解烃工程菌

用供体菌C_(20-11)和D_(36-3)染色体DNA转化受体菌T_3原生质球。采用5mg/ml溶菌酶,37℃水浴1小时制备原生质球。原生质球形成率为99％,再生率为72.5％。在30％、PEG6000和0.1MCaC1_2作用下,供体DNA转化受体原生质球。我们用上述条件构建了能够同时降解以下四种碳源:苯甲酸、萘、樟脑、十六烷的工程菌株TCD32-14-1。     

珠蒜苞鳞衰退组织中原生质撤离的两条途径的电子显微镜观察

高等植物衰退组织内的半解体原生质,可通过胞间运动转移出去提供给正在兴旺生长的组织。娄成后等曾在大蒜鳞片等材料中应用光学显微镜观察到衰退组织的原生质撤离甚至入侵导管的现象。我们用电子显微镜对上述现象作了进一步观察,观察到原生质撤离的胞间运动以及半解体的原生质进入细胞间隙和导管。     

原生质流动的观察实验

近来我用孔雀草(万寿菊Tagetes patula)花的表皮毛观察有色体时,同时发现了原生质的流动。经与其它材料的比较,我觉得用孔雀草花的表皮毛观察原生质流动效果较好。且具有取材方便、操作简易、杂色体多、原生质流动快等特点,便于学生掌握。     

浙贝母鳞片衰退和物质撤退的超微结构观察

本文以浙贝母衰退鳞片为材料,观察了细胞内含物降解和运输的过程。细胞内含物降解时,内膜系统产生许多囊泡,这些囊泡有降解和运输两方面的作用。降解产物在细胞中表现为颗粒和丝状物形式,它们在细胞间转移通过共质体和质外体两条途径,转移方式多种多样．降解产物经过细胞间转移,最终汇集到维管束,再通过维管束运往新生器官。转移细胞在物质运往筛分子的过程中起着重要作用。韧皮部是降解产物运输的主要通道,导管的一部分可能也参与了这种运输．     

荔枝雄花性别决定过程中细胞超微结构的变化

荔枝雄花雌蕊原基在大孢子母细胞减数分裂后开始衰退.内质网历经增生扩展,穿壁相连,同心缠绕,多条平行弯曲,不规则堆叠.内质网和高尔基体产生许多囊泡,囊泡在细胞内含物的降解和运输过程中起着重要的作用.线粒体在雌蕊原基细胞衰败的前、中期数量增加,后期分批降解.过氧化物酶体在雌蕊原基细胞衰败的中期紧挨核短暂出现.细胞核的染色质凝集断裂;核周腔扩大,形成胀泡;染色质趋边,外泄.细胞原生质表现出有序的、在膜包裹下的降解,首先是核糖体,而后依次是:过氧化物酶体、内质网、高尔基体、线粒体、核.雌蕊原基的衰败历程可能是一种程序性细胞死亡的过程.     

Chemically induced degradation of CK2 by proteolysis targeting chimeras based on a ubiquitin–proteasome pathway

As a ubiquitous, highly pleiotropic and constitutively active serine/threonine protein kinase, casein kinase 2 (CK2) is closely associated with tumorigenesis by its overexpression in cancer cells. Here we report several proteolysis targeting chimeras (PROTACs) via “click reaction” to connect a CK2 inhibitor (CX-4945) and pomalidomide for degradation of CK2 protein. Among them, compound 2 degraded CK2 in a dose and time-dependent manner, and kept CK2 at a low basal level by recruiting ubiquitin-proteasome system. The degradation of CK2 resulted in the reduced phosphorylation of Akt and the up-regulation of p53. As a CK2 protein degrader, 2 showed the analogous cytotoxicity to CX-4945 but with a quite different mechanism of action from the CK2 inhibitor, hinting that degradation of CK2 proteins by PROTACs is a potential way for cancer treatments.     

Low Cell Number Proteomic Analysis Using In-Cell Protease Digests Reveals a Robust Signature for Cell Cycle State Classification

Comprehensive proteome analysis of rare cell phenotypes remains a significant challenge. We report a method for low cell number MS-based proteomics using protease digestion of mildly formaldehyde-fixed cells in cellulo, which we call the “in-cell digest.” We combined this with averaged MS1 precursor library matching to quantitatively characterize proteomes from low cell numbers of human lymphoblasts. About 4500 proteins were detected from 2000 cells, and 2500 proteins were quantitated from 200 lymphoblasts. The ease of sample processing and high sensitivity makes this method exceptionally suited for the proteomic analysis of rare cell states, including immune cell subsets and cell cycle subphases. To demonstrate the method, we characterized the proteome changes across 16 cell cycle states (CCSs) isolated from an asynchronous TK6 cells, avoiding synchronization. States included late mitotic cells present at extremely low frequency. We identified 119 pseudoperiodic proteins that vary across the cell cycle. Clustering of the pseudoperiodic proteins showed abundance patterns consistent with “waves” of protein degradation in late S, at the G2&M border, midmitosis, and at mitotic exit. These clusters were distinguished by significant differences in predicted nuclear localization and interaction with the anaphase-promoting complex/cyclosome. The dataset also identifies putative anaphase-promoting complex/cyclosome substrates in mitosis and the temporal order in which they are targeted for degradation. We demonstrate that a protein signature made of these 119 high-confidence cell cycle–regulated proteins can be used to perform unbiased classification of proteomes into CCSs. We applied this signature to 296 proteomes that encompass a range of quantitation methods, cell types, and experimental conditions. The analysis confidently assigns a CCS for 49 proteomes, including correct classification for proteomes from synchronized cells. We anticipate that this robust cell cycle protein signature will be crucial for classifying cell states in single-cell proteomes.     

Aerobic batch degradation of phenol using immobilized Pseudomonas putida

Alginate concentrations between 2 and 4% had little effect on the degradation rate of phenol by alginate-immobilized Pseudomonas putida. Ten-degree shifts from 25°C resulted in approximately 30% slower degradation. Maximal degradation rates were favored at pH 5.5–6.0. The response of degradation rate to increased air flow in the bubble column used was almost linear and an optimal higher than 16 vol vol⁻¹ was indicated, although free cells appeared in the reaction medium above 12 vol vol⁻¹. When the initial phenol concentration was raised, degradation rate was not significantly affected until levels higher than 1200 mg ml⁻¹ where performance was markedly reduced. Increasing the ratio of total bead volume to medium volume gave progressively smaller increases in degradation rate. At a medium volume to total bead volume ratio of 5:1, the maximum degradation rate was 250 mg L⁻¹ h⁻¹. Received 24 November 1998/ Accepted in revised form 27 January 1999     

KIAA1363 affects retinyl ester turnover in cultured murine and human hepatic stellate cells

Large quantities of vitamin A are stored as retinyl esters (REs) in specialized liver cells, the hepatic stellate cells (HSCs). To date, the enzymes controlling RE degradation in HSCs are poorly understood. In this study, we identified KIAA1363 (also annotated as arylacetamide deacetylase 1 or neutral cholesterol ester hydrolase 1) as a novel RE hydrolase. We show that KIAA1363 is expressed in the liver, mainly in HSCs, and exhibits RE hydrolase activity at neutral pH. Accordingly, addition of the KIAA1363-specific inhibitor JW480 largely reduced RE hydrolase activity in lysates of cultured murine and human HSCs. Furthermore, cell fractionation experiments and confocal microscopy studies showed that KIAA1363 localizes to the endoplasmic reticulum. We demonstrate that overexpression of KIAA1363 in cells led to lower cellular RE content after a retinol loading period. Conversely, pharmacological inhibition or shRNA-mediated silencing of KIAA1363 expression in cultured murine and human HSCs attenuated RE degradation. Together, our data suggest that KIAA1363 affects vitamin A metabolism of HSCs by hydrolyzing REs at the endoplasmic reticulum, thereby counteracting retinol esterification and RE storage in lipid droplets.     

Methods for determining the efficacy of radical-trapping antioxidants

Hydrocarbon autoxidation is the free radical chain reaction primarily responsible for the oxidative degradation of organic materials, including those that make up cells, tissues, and organs. The identification of compounds that slow this process (antioxidants) and the quantitation of their efficacies have long been goals of academic and industrial researchers. Antioxidants are generally divided into two types: preventive and radical-trapping (also commonly referred to as chain-breaking). Preventive antioxidants slow the rate of initiation of autoxidation, whereas radical-trapping antioxidants slow the rate of propagation by reacting with chain-propagating peroxyl radicals. The purpose of this review is to provide a comprehensive overview of different approaches to measure the kinetics of the reactions of radical-trapping antioxidants with peroxyl radicals, and their use to study the inhibition of hydrocarbon (lipid) autoxidation in homogeneous solution, as well as biphasic media (lipid bilayers) and cell culture. Direct and indirect approaches are presented and advantages and disadvantages of each are discussed in order to facilitate method selection for investigators seeking to address particular questions in this immensely popular field.     

Germinal proto-mitochondria from rat liver

A number of properties of the smallest (less than 0.2 μm) germinal proto-mitochondria (PRMC) from rat liver have been studied. These PRMC were obtained by filtering the light fraction of hepatic mitochondria (MC) through calibrated millipore membranes. Germinal PRMC contain in general the same proteins as MC. However, they have the reduced content of flavoproteins and zero cytochrome oxidase. Germinal PRMC, in contrast to MC, almost does not contain the “aging pigment” - lipofuscin. They have DNA; the DNA/protein ratio in them is much higher than in MC, i.e. they are poor in protein. The obtained results support the earlier assumption that MC in specialized animal cells can arise from germinal PRMC - particles smaller than 0.2 μm containing DNA. It is assumed that the DNA molecules enter to cytoplasm during degradation of old MC serves as a seed for the formation of PRMC (with the connection of nuclear DNA).     

The Acyl-Proteome of Syntrophus aciditrophicus Reveals Metabolic Relationships in Benzoate Degradation

Syntrophus aciditrophicus is a model syntrophic bacterium that degrades fatty and aromatic acids into acetate, CO₂, formate, and H₂ that are utilized by methanogens and other hydrogen-consuming microbes. S. aciditrophicus benzoate degradation proceeds by a multistep pathway with many intermediate reactive acyl-coenzyme A species (RACS) that can potentially N^ε-acylate lysine residues. Herein, we describe the identification and characterization of acyl-lysine modifications that correspond to RACS in the benzoate degradation pathway. The amounts of modified peptides are sufficient to analyze the post-translational modifications without antibody enrichment, enabling a range of acylations located, presumably, on the most extensively acylated proteins throughout the proteome to be studied. Seven types of acyl modifications were identified, six of which correspond directly to RACS that are intermediates in the benzoate degradation pathway including 3-hydroxypimeloylation, a modification first identified in this system. Indeed, benzoate-degrading enzymes are heavily represented among the acylated proteins. A total of 125 sites were identified in 60 proteins. Functional deacylase enzymes are present in the proteome, indicating a potential regulatory system/mechanism by which S. aciditrophicus modulates acylation. Uniquely, N^ε-acyl-lysine RACS are highly abundant in these syntrophic bacteria, raising the compelling possibility that post-translational modifications modulate benzoate degradation in this and potentially other, syntrophic bacteria. Our results outline candidates for further study of how acylations impact syntrophic consortia.     

Melanin endocytosis by cultured mammalian cells : A model for melanin in a cellular environment

The incorporation of natural eumelanin from bovine eyes and synthetic 3,4-dihydroxy-phenylalanine (dopa) melanin into Chinese hamster ovary (CHO) cells is reported. The process is linear for at least 8 h. Electron microscopy showed phagocytosis of melanin, either as a single granule or in groups of granules, into cell lysosomes with subsequent degradation of the granule. The general features of the ingestion and degradation processes mimic those of the incorporation of melanosomes into keratinocytes. CHO cells with ingested melanin in general revealed properties very similar to those of the pigment-free CHO cell: cell division, oxygen consumption and plating efficiency were not greatly altered by moderate concentrations of pigment. This suggests that the CHO cell system may be useful for the study of pigment in a cellular environment; pigment-free CHO cells are well characterized and can serve as a good control. Preliminary applications are reported: demonstrations of (1) incorporation of metal ions (Al3+) into CHO cells using melanin as a carrier; (2) the ability of melanin to enhance the rate of oxygen consumption during photo-irradiation of the cells.     

Compatibility of PTB7 and [70]PCBM as a Key Factor for the Stability of PTB7:[70]PCBM Solar Cells

The rapid degradation of organic photovoltaic (OPV) devices compared to conventional inorganic solar cells is one of the critical issues that have to be solved in order to make OPV a competitive commercial technology. The understanding of the fundamental mechanisms that reduce the power conversion efficiency (PCE) over time is beneficial for the design of new materials with enhanced stability. This paper focuses on bulk heterojunction organic solar cells based on thieno [3,4‐b] thiophene‐alt‐benzodithiophene (PTB7) mixed with [6,6]‐phenyl‐C71‐butyric acid methyl esther ([70]PCBM). In spite of being promising in terms of PCE, devices based on this blend are unstable and have a short lifetime. When exposed to light in inert atmosphere, the PCE drops by 15% in less than 1 h and by 35% in 8 h; this degradation is induced by the ultraviolet (UV) part of the spectrum. This paper analyzes the effect induced by UV light on the transport of charges in PTB7:[70]PCBM. Contrary to expectations, the electron transport shows evidence of trapping, while the transport of holes appears unaffected. Furthermore, it is proven that the loss of PCE is due to a reaction between PTB7 and [70]PCBM, while the intrinsic instability of the polymer plays a marginal role.