大豆根瘤侵染细胞发育中高尔基体的变化

为了探讨高尔基体与细菌增殖和周膜扩展的关系,用电子显微镜研究了大豆根瘤侵染细胞发育中高尔基体的变化.结果表明:幼龄侵染细胞中有高尔基体,但在成熟和衰老的侵染细胞中却几乎没有这种结构.高尔基体随幼龄侵染细胞中细菌的增多而增多,它们主要位于细菌附近.这些高尔基体的生理活性较高,不断向周围的细胞质分泌含有纤维状物质的小泡,有的小泡还在向细菌运动.一些小泡位于细菌的周膜上,其附近细菌的周膜常向外形成隆起,并有纤维状物质出现在其中.由此说明,大豆根瘤侵染细胞中的高尔基体参与了细菌的增殖和周膜的扩展.     

Induction of apoptosis and non-apoptosis in human breast cancer cell line (MCF-7) by cisplatin and caffeine

Background: Molecular targeted therapy by different cell death inducers are recently considered in cancer therapy. The aim of this study was to compare the effect of cisplatin and inositol trisphosphate kinase inhibitor (caffeine) on human breast cancer cell line (MCF-7). The pattern of cell death in MCF-7 cells following the exposure to cisplatin and caffeine in individual and combination forms was characterized. Methods: MCF-7cells at late exponential phase were divided into two groups: control and experimental groups. Experimental group was exposed to cisplatin, caffeine and combination of them and control group was treated by vehicle. Forty-eight hours after incubation, floating and attached cells were collected separately. Flowcytometry analysis and electron microscopy were carried out on both attached and floating cells. Results: Two types of apoptotic and non-apoptotic cells were observed in the floating cells as well as in sub G1 cells of both experimental and control groups by electron microscopy. Both early and late stages of apoptosis were characterized and the attached cells remained unaffected. Conclusion: Although two different forms of cell death (apoptosis and non-apoptosis) were appeared in MCF-7 following exposure to cisplatin and caffeine, apoptosis was the major mechanism of cell death. The combination form of anti-cancer drugs with different mechanisms could decrease the dosage of employed anti-cancer drugs.Key Words: Cisplatin, Caffeine, Apoptosis     

The cellular structure of cork from Quercus cerris var. cerris bark in a materials’ perspective

Cork in the outer bark of trees is among the valuable raw materials of biological origin due to properties that result mainly from its cellular structure. Large scale commercial utilization of cork has been only achieved with cork from Quercus suber. Another oak species, Quercus cerris, also contains substantial, albeit not continuous, regions of cork that are clearly visible to the naked eye but are so far considered as a waste material.Bark samples of Q. cerris var. cerris trees were collected from the And?r?n province, Turkey. Cork portions were separated and their cellular structure was investigated with optical and electron scanning microscopy observations. The results were compared with Q. suber cork.Q. cerris cork has the typical features of cork tissues with a regular and radially aligned structure of suberized cells without intercellular voids, showing a ring structure and a distinction of earlycork and latecork cells. Solid volume fraction was estimated at 25% (22% in earlycork, 36% in latecork).In Q. cerris cork cells are smaller, cell wall thickness and solid volume fraction are higher, and the tissue is less homogeneous with a higher content of lignified inclusions than in Q. suber cork. These factors will negatively influence quality in regard to density and mechanical properties associated to elasticity. However, this does not impair its use for production of granulates and agglomerates, e.g. for insulation and energy absorption. Separation of the cork fraction from the bark is a step required before further processing and use.     

胶孢炭疽菌侵染芒果的过程及寄主的组织病理学研究

炭疽菌是一类地理分布广泛、寄主范围广泛的真菌,在作物成熟前和成熟后都能侵染植株和果实,特别对成熟的热带作物危害极大,造成果实腐烂,导致严重的经济损失并影响产品的出口质量.为探讨胶孢炭疽菌侵染芒果的过程及其与寄主的互作关系,揭示该病菌对果实和叶片的侵染方式和扩展途径,以元江本地品种'三年芒'病果上分离、筛选的强致病菌株C...     

Investigation of Host Responses of Different Potato Genotypes at Tissue, Cellular and Subcellular Levels After Infection with Phytophthora infestans

In histological and cytological investigations, the infection process of Phytophthora infestans, the late blight pathogen, was comparatively studied in several potato cultivars and somatic hybrid genotypes and their parents using fluorescence microscopy and electron microscopy methods. The results showed that germination of zoospores of P. infestans and frequency of invading by infection hyphae did not differ among the cultivar-pathogen interactions, but, extension of hyphae in host cells markedly differed among these genotypes. In the susceptible genotypes the pathogen grew rapidly inter- and intracellularly, 12 h after inoculation (hai), and some digital like haustoria were formed and the cytoplasm of the host cells became disorganized. In the resistance genotypes, the pathogen was restricted to the site of initial penetration, although some hyphae could penetrate the epidermal cell, however, the host cells produced resistance responses, such as formed wall appositions when in contact with hyphae, and no haustoria like structures were found. In the somatic hybride genotypes, the host response was different according to their parents as shown by transmission electron microscopy. In the hybrid genotype 1508/2, like in the wild species S. bulbocastanum, no hyphae were found in host cells. In the other genotypes, hyphae of P. infestans spread intercellularly and formed haustoria, but the cytoplasm of hyphae and haustoria was disorganized and host cell resistant responses often appeared, such as, host cells were disorganized and necrotic and cell wall apposition were observed.     

VSPs在热带树木中分布的细胞学研究

采用光学和电子显微镜技术研究VSPs在18科54属70种热带树木中的分布和超微结构。VSPs广泛分布于热带树木中。含羞草科、苏木科和蝶形花科树木体内普遍存在VSPs,而且落叶树种明显比常绿树种丰富。大戟科只有少数属树种有VSPs。VSPs在同一属树种之间的分布是一致的。在树木体内,VSPs主要分布在树皮组织,尤其是次生韧皮部中。这些蛋白质积累在韧皮薄壁细胞和韧皮射线细胞的中央大液泡里。VSPs的形态多样,不同形态的VSPs分别存在于不同的细胞中。VSPs的分布和形态的多样性是树木的一个分类特征。      

Intrabody Tetrodotoxin Distribution and Possible Hypothesis for Its Migration in Ribbon Worms Cephalothrix cf. simula (Palaeonemertea,Nemertea)

Tetrodotoxin (TTX) is a potent neurotoxin found in many marine and terrestrial animals, but only a few species, such as the ribbon worms of the genus Cephalothrix, accumulate it in extremely high concentrations. The intrabody distribution of TTX in highly toxic organisms is of great interest because it helps researchers to understand the pathways by which the toxin migrates, accumulates, and functions in tissues. Using immunohistochemistry with anti-TTX antibodies, the authors of this study investigated the toxin’s distribution inside the organs, tissues, and cells of Cephalothrix cf. simula. The cell types of TTX-positive tissues were identified by light microscopy. The main sites of TTX accumulation occurred in the secretory cells of the integuments, the microvilli of the epidermal ciliary cells, cephalic glands, the glandular epithelia of the proboscises, the enterocytes of the digestive systems, and nephridia. Obtained data suggest the toxin migrates from the digestive system through blood vessels to target organs. TTX is excreted from the body through the nephridia and mucus of epidermal cells.     

无患子科树木细子龙营养贮藏蛋白质的分离鉴定

采用光学显微技术、十二烷基磺酸钠-聚丙稀酰胺凝胶电泳(SDS—PAGE)、免疫印迹和间接免疫荧光定位技术，分离鉴定了无患子科树木细子龙(Amesiodendron chinense)的营养贮藏蛋白质(VSP)。在经汞-溴酚蓝染色的石蜡切片中，观察到枝条、树干和根的次生木质部的木薄壁细胞中有深蓝色颗粒状物质，而在上述枝条、树干和根的次生韧皮部的韧皮薄壁细胞和韧皮射线薄壁细胞中则为均一状的淡蓝色物质。这些被染成不同程度蓝色的物质具有蛋白质性质。SDS—PAGE分析表明，在枝条、树干和根的树皮及木质部中，有2种含量高的蛋白质，其分子质量约23kDa和26kDa。使用荔枝(一种无患子科树木)的22kDa VSP的多克隆抗体对这些部位的树皮和木质部可溶性蛋白质进行免疫印迹分析，在免疫印迹图谱上，该抗血清只识别23kDa和26kDa蛋白质，且23kDa蛋白质谱带颜色较深，说明它们与荔枝的22kDa VSP有一定的同源性。间接免疫荧光定位结果表明，只有颗粒状和均一状蛋白质内含物发出异硫氰酸荧光素特异的蓝绿色荧光，说明23kDa和26kDa蛋白质是这些颗粒均一状内含物的主要成分。树木抽新梢后，这2种蛋白质含量在一定程度上减少。可见，23kDa和26kDa蛋白质符合树木VSP的3条标准，应该是细子龙的VSP。韧皮部和木质部的VSP在形态上的差异可能与蛋白质的种类不同有关。     

In vitro observation of dynamic ordering processes in the extracellular matrix of living, adherent cells

Collecting information at the interface between living cells and artificial substrates is exceedingly difficult. The extracellular matrix (ECM) mediates all cell-substrate interactions, and its ordered, fibrillar constituents are organized with nanometer precision. The proceedings at this interface are highly dynamic and delicate. In order to understand factors governing biocompatibility or its counterpart antifouling, it is necessary to probe this interface without disrupting labels or fixation and with sufficient temporal resolution. Here the authors combine nonlinear optical spectroscopy (sum-frequency-generation) and microscopy (second-harmonic-generation), fluorescence microscopy, and quartz crystal microgravimetry with dissipation monitoring in a strategy to elucidate molecular ordering processes in the ECM of living cells. Artificially (fibronectin and collagen I) and naturally ordered ECM fibrils (zebrafish, Danio rerio) were subjected to nonlinear optical analysis and were found to be clearly distinguishable from the background signals of diffusive proteins in the ECM. The initial steps of fibril deposition and ordering were observed in vitro as early as 1 h after cell seeding. The ability to follow the first steps of cell-substrate interactions in spite of the low amount of material present at this interface is expected to prove useful for the assessment of biomedical and environmental interfaces.     

Comparing invasive and non-invasive of isolated Shigella flexneri by electron microscopy of cell culture, SDS-PAGE and Congo red method

BACKGROUND: The aim of this study was to compare invasive and non-invasive strains of Shigella flexneri isolated from Tehran by a 120 kDa protein band by SDS-PAGE, electron microscopy of cell culture and Congo red dye methods. METHODS: S. flexneri strains were isolated by standard bacterial methods from fecal specimens of children attending to the 3 children's hospitals. Phenotype analysis for screening virulent of strains of S. flexneri was done on a plate of tryptic soy agar contained 0.003% Congo red dye. Whole membrane protein preparations were used to examine the protein profiles of the inner and outer membrane of these Gram-negative bacteria. The protein mixture was electrophoresed through a polyacrylamide gel. The gel was stained with Coomassie brilliant blue R250 and destained with ethanol and acetic acid. HeLa cell culture was done by two-step preparations: one for light microscopy and the other for electron microscopy. RESULTS: Some of S. flexneri (46%) were Congo red positive colonies. S. flexneri with negative Congo red phenotype could not enter the HeLa cell culture. A 120 kDa protein band was found in 46% of these bacteria which could enter into HeLa cell culture. Pseudopod structures which facilitate bacterial cell-to-cell spread were readily identified by electron microscopy. DISCUSSION: Since the existence of 120-kDa protein band was corresponded to enter of S. flexneri into the HeLa cell culture and correlated with Congo red dye positive, for identification of invasive and non-invasive S. flexneri strains, the use of a 120-kDa protein band by SDS-PAGE or a simple, rapid and very cheap Congo red dye method is recommended. Because, there are some deaths due to Shigella sp. in our country, notification on the isolation of these bacteria in both children hospitals laboratories and private clinical laboratories is important.     

Various nano-silica particles affecting dyeability of poly(ethylene terephthalate)/silica nanocomposite films

Poly(ethylene terephthalate) [PET] based nanocomposites containing three differently modified silica particles were prepared by melt compounding. The influence of type of nano-silica on dispersibility, thermal and dyeing properties of the resultant nanocomposite was investigated by various analytic techniques, namely, polarized optical microscopy (POM), scanning electron microscopy (SEM), differential scanning calorimetry (DSC), thermal gravimetric analysis (TGA), reflectance spectroscopy (RS), and light fastness. Optical microscopy images illustrated that nano-silica particles tended to increase the number of spherulites in the PET matrix which were dependent on nano-silica type and content. Thermal studies of the resultant nanocomposites showed a slight decrease in the melting temperature compared to a pristine PET. Silica nanocomposites were finally dyed with a disperse dye and their reflectances were determined by the aid of reflectance spectrophotometer. Such reflectances were converted to the corresponding color coordinate values which are indicative of dyeability of such nanocomposites.     

Fibroblast adhesion on unidirectional polymeric nanofilms

Nanotextured polymeric surfaces with inclined rods reveal highly anisotropic properties concerning wetting and adhesion. In this work, we report on the interaction of fibroblast cells with these highly anisotropic materials. The authors quantified removal of adherent cells from such surfaces by a laminar flow. The critical shear force needed for cell removal from the surface depends on the inclination direction. Based on electron microscopy cross sections we deduce that interactions of cellular filopodia extending into the nanotextured surface are causing the direction depending removal.     

Characterization of anatomy, ultrastructure and lignin microdistribution in Forsythia suspensa

Forsythia suspensa, as a traditional Chinese medicinal herb, has been studied extensively. In this work, morphological and anatomical features of F. suspensa stem were examined by light microscopy (LM) and scanning electron microscopy (SEM). The results showed that the slenderness and Runkel ratio is 51 and 0.67, respectively. Anatomical observations indicated that F. suspensa is diffuse-porous wood. Helical thickenings and alternate intervessel pits are present on vessel cell wall. Transmission electron microscopy (TEM) images exhibited that the fiber cell wall is typically differentiated into three layers: middle lamella (ML), primary wall (P) and secondary wall (S₁, S₂ and S₃), and the staining intensities represent differing lignin concentrations. Also, the confocal laser scanning microscopy (CLSM) and scanning electron microscopy with energy dispersive X-ray analysis (SEM-EDXA) were used to investigate the lignin distribution in cell wall qualitatively and semi-quantitatively. Confocal images (488 nm) revealed a high level of lignin autofluorescence in the cell corner middle lamella (CCML), with lower levels of fluorescence in the compound middle lamella (CML) and S₂ region. The results from SEM-EDXA demonstrated that lignin concentration ratio in different regions of fiber wall is 1.3 (CCML):1.1 (CML):1 (S₂).     

花生感染条纹病毒(PStV)后叶片细胞超微结构研究

花生接种PStV后30d取叶片进行超薄切片置于电镜下观察,在花37和白沙1016两个品种发病叶片的叶肉细胞中均发现有多种形状的内含体和病毒粒子的存在;内含体多发现于细胞核周围,有风轮状、柬状、管状和环状等;病毒粒子以柬状或拟晶格状存在,且以高感品种白沙1016叶肉细胞内的含量较多。PStV对叶绿体和线粒体等细胞器的结构有破坏作用,表现为叶绿体畸形,类囊体片层减少,基粒缩小且排列不整齐,并有大量淀粉粒累积,部分线粒体出现肥大和自溶。此外,感病细胞的膜系统松弛,有膜溶解现象,上述病理结构变化均以高感品种白沙1016表现的更为明显。     

乙烯丰诱导橡胶树对条溃疡病抗性的超微结构

用透射电镜技术研究了柑桔褐腐疫霉(Phytophthora citrophthora)引起的橡胶树(Hevea brasiliensis Mull.Arg.)无性系 RRIM600条溃疡病,比较了正常树皮(即不施乙烯丰树皮)和施用乙烯丰(2—氯乙基磷酸,乙烯释放剂)树皮接种后菌丝的侵染和寄主细胞的反应。在正常树皮中,菌丝主要在寄主细胞间生长并引起细胞壁分解。菌丝也常常垂直穿透寄主细胞壁而在细胞内形成吸器和细胞内菌丝。当开始垂直穿透时,寄主细胞壁常常形成乳头状突起。吸器具有吸器外基质(extrahaustorial matrix),也常常具有领(collar)。施用乙烯丰的树皮病斑比正常树皮上的较早停止扩展而且病斑小得多,但是未发现菌丝侵染和寄主细胞反应的方式有明显差别。据用商品纤维素酶和果胶酶对树皮细胞壁试验,乙烯丰处理的树皮比正常树皮较能抵抗这些酶的分解。这些事实表明,乙烯丰诱导的寄主细胞的变化使菌丝侵染延迟,但并未杀伤病原,也没有完全阻止穿透。     

Hardness Distribution and Endosperm Structure on Polishing Characteristics of Brewer’s Rice Kernels

Abstract

This study was designed to determine the effects of the hardness distribution and the endosperm structure on the polishing characteristics of brewer’s rice kernels. We used four brewer’s rice cultivars, Kairyo-omachi, Hattan-nishiki No. 1, Senbon-nishiki and Yamada-nishiki. The broken kernel ratios in Kairyo-omachi and Hattan-nishiki No. 1 were significantly higher than those in Senbon-nishiki and Yamada-nishiki. Vickers hardness (VH) values in white-core tissues in kernels differed among varieties, which were significantly lower in Kairyo-omachi and Hattan-nishiki No. 1. However, no varietal differences were observed in VH values in the peripheral translucent tissues surrounding the white-core tissues. The tissues along the dorsoventral axis were softer than those along the longitudinal axis of the kernels. The tissues on the ventral side were softer than those on the dorsal side. Scanning electron microscopy (SEM) observations revealed the presence of closely arranged compound starch granules and few varietal differences in the peripheral translucent tissues surrounding the white-core tissues. However, as compared with Yamada-nishiki and Senbon-nishiki, in Hattan-nishiki No. 1 and Kairyo-omachi, the starch granules were loosely packed and the airspaces between the starch granules were more numerous in the white-core tissues. A higher number of airspaces and less starch were present in the endosperm cells along the dorsoventral axis when compared with along the longitudinal axis and on the ventral side than on the dorsal side. The present study showed that polishing characteristics are closely related with the endosperm structure, which is characterized as the density of starch granules.     

The Role of Biodegradable Engineered Nanofiber Scaffolds Seeded with Hair Follicle Stem Cells for Tissue Engineering

Background

The aim of this study was to fabricate the poly caprolactone (PCL) aligned nanofiber scaffold and to evaluate the survival, adhesion, proliferation, and differentiation of rat hair follicle stem cells (HFSC) in the graft material using electrospun PCL nanofiber scaffold for tissue engineering applications.

Methods

The bulge region of rat whisker was isolated and cultured in DMEM: nutrient mixture F-12 supplemented with epidermal growth factor. The morphological and biological features of cultured bulge cells were observed by light microscopy using immunocytochemistry methods. Electrospinning was used for production of PCL nanofiber scaffolds. Scanning electron microscopy (SEM), 3-(4, 5-di-methylthiazol- 2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay, and histology analysis were used to investigate the cell morphology, viability, attachment and infiltration of the HFSC on the PCL nanofiber scaffolds.

Results

The results of the MTT assay showed cell viability and cell proliferation of the HFSC on PCL nanofiber scaffolds. SEM microscopy images indicated that HFSC are attached, proliferated and spread on PCL nanofiber scaffolds. Also, immunocytochemical analysis showed cell infiltration and cell differentiation on the scaffolds.

Conclusion

The results of this study reveal that PCL nanofiber scaffolds are suitable for cell culture, proliferation, differentiation and attachment. Furthermore, HFSC are attached and proliferated on PCL nanofiber scaffolds.Key Words: Nanofiber, Electrospinning, Stem cells, Tissue engineering     

Effects of High Temperature Stress on Microscopic and Ultrastructural Characteristics of Mesophyll Cells in Flag Leaves of Rice

The microscopic and ultrastructural characteristics of mesophyll cells in flag leaves of two rice lines (a thermo-sensitive line 4628 and a thermo-resistant line 996) under high temperature stress (37°C during 8:00-17:00 and 30°C during 17:00-8:00) were investigated using an optical and a transmission electron microscopy. The membrane permeability and malondialdehyde content increased under the high temperature stress, and the increase of both variables was greater in the line 4628 than in the line 996. Und...     

巨型线粒体形成机理的探讨

我们用透射电镜观察了大豆根瘤中巨型线粒体的形成。在细菌侵染初期，寄主细胞中的线粒体一般呈随机分布，然后逐渐移向细胞壁，常常位于造粉体周围。进而体积变大，相互接触，发生融合。变为一个较大的线粒一。当这种线粒体进一步相互融合时，最后就形成了一种十分罕见的巨型线粒体。由于它是线粒体相互融合而成，因此往往留下一些融合痕迹。     

Some anatomical and physiological potato tuber characteristics and their relationship to hollow heart

The inception site of hollow heart (HH) and possible relationships between HH and physiological or anatomical characteristics of tubers were investigated. Scanning electron micrographs revealed cells at the site of HH inception were physically rather than enzymatically degraded. Transmission electron micrographs of tuber pith cells from non hollow (NH) tubers showed cell wall and cytoplasm width varied among cultivars but were not related to HH resistance. Water potential of NH tuber tissues at the pith, bud, center and stem end of 3 cultivars and one selection varying in resistance to HH was determined. Although the water potential gradually became less negative as tubers advanced in maturity, no differences were found in water potential between pith cells in various parts of tubers or among cultivars. Tuber pith cells increased in size concomitant with tuber growth. In all tubers pith cells were largest in the center and stem end and smallest in the bud end, but differences in mean pith cell size among cultivars were not related to HH susceptibility. However, within a cultivar the bud, center, and stem end pith cells of HH tubers were all smaller than the corresponding pith cells from similarly sized NH tubers.