Germination of Phyllosticta ampelicidaPycnidiospores: Prerequisite of Adhesion to the Substratum and the Relationship of Substratum Wettability

Pycnidiospores of Phyllosticta ampelicida, the causal agent of black rot of grape, were found to germinate only on substrata on which they were firmly attached. Such surfaces were poorly wettable and had advancing contact angles (straight thetaa) formed by a water drop of >80°, e.g., grape leaf, polystyrene, Teflon, polycarbonate, collodion, and glass treated with the silanes n-octadecyltrichlorosilane, dimethyldichlorosilane, or diphenyldichlorosilane. When pycnidiospores were deposited on more wettable surfaces they did not attach firmly and did not germinate. Such highly wettable surfaces had straight thetaa 相似文献   

A Role for Topographic Cues in the Organization of Collagenous Matrix by Corneal Fibroblasts and Stem Cells

Human corneal fibroblasts (HCF) and corneal stromal stem cells (CSSC) each secrete and organize a thick stroma-like extracellular matrix in response to different substrata, but neither cell type organizes matrix on tissue-culture polystyrene. This study compared cell differentiation and extracellular matrix secreted by these two cell types when they were cultured on identical substrata, polycarbonate Transwell filters. After 4 weeks in culture, both cell types upregulated expression of genes marking differentiated keratocytes (KERA, CHST6, AQP1, B3GNT7). Absolute expression levels of these genes and secretion of keratan sulfate proteoglycans were significantly greater in CSSC than HCF. Both cultures produced extensive extracellular matrix of aligned collagen fibrils types I and V, exhibiting cornea-like lamellar structure. Unlike HCF, CSSC produced little matrix in the presence of serum. Construct thickness and collagen organization was enhanced by TGF-ß3. Scanning electron microscopic examination of the polycarbonate membrane revealed shallow parallel grooves with spacing of 200–300 nm, similar to the topography of aligned nanofiber substratum which we previously showed to induce matrix organization by CSSC. These results demonstrate that both corneal fibroblasts and stromal stem cells respond to a specific pattern of topographical cues by secreting highly organized extracellular matrix typical of corneal stroma. The data also suggest that the potential for matrix secretion and organization may not be directly related to the expression of molecular markers used to identify differentiated keratocytes.     

Multilamellar Vesicular Clusters of Phosphatidylcholine and Their Sensitivity to Spectrin: A Study by Fractal Analysis

The cluster patterns of multilamellar vesicles (MLV) of dimyristoylphosphatidylcholine (DMPC) were analyzed using a combination of fractal analysis and lattice simulation. Self-assembly of DMPC MLVs resulted in two types of microscopically observable clusters. The clusters were classified on the basis of their mass fractal dimension, two-dimensional porosity, and the light scattering properties. Spectrin, a cytoskeletal protein, well known for its role in determining the cellular morphology, was used to perturb such spontaneously formed clusters. The fragmentation of the clusters by hydrodynamic perturbation followed a power law, implying again a fractal behavior. A lattice-based simulation was performed generating different class of cluster patterns. The observed correspondence between the cluster patterns and their stability was discussed in the framework of the proposed lattice simulation.     

Cytotoxicity of Adenovirus-Antibody Aggregates: Sensitivity to Different Cell Strains, and Inhibition by Hexon Antiserum and by Complement

Adenovirus-antibody aggregates under defined conditions are cytotoxic in vitro. All members of adenovirus groups I, II, and III caused toxicity upon aggregation. The toxicity of the clusters is exerted by the virions. Toxicity is temperature dependent and may be caused by a mechanism similar to that used in viral penetration. Cells permitting direct viral penetration were all sensitive to the toxic aggregates. The toxicity seems to be related to hexon antigens on the surface of the virions since antihexon sera neutralized the toxicity. No evidence was obtained showing that pentons are required for this kind of cytotoxicity. Adenovirus types 3, 5, and 9 were used in the experiment. Cytotoxicity was estimated by the (51)Cr release assay. Complement factors could be excluded as mediators of the cytolytic reactions. Instead, complement was shown to prevent the formation of toxic aggregates or to neutralize the toxicity of preformed ones.     

Exosomes Derived from Human Primary and Metastatic Colorectal Cancer Cells Contribute to Functional Heterogeneity of Activated Fibroblasts by Reprogramming Their Proteome

Cancer‐associated fibroblasts (CAFs) are a heterogeneous population of activated fibroblasts that constitute a dominant cellular component of the tumor microenvironment (TME) performing distinct functions. Here, the role of tumor‐derived exosomes (Exos) in activating quiescent fibroblasts into distinct functional subtypes is investigated. Proteomic profiling and functional dissection reveal that early‐ (SW480) and late‐stage (SW620) colorectal cancer (CRC) cell‐derived Exos both activated normal quiescent fibroblasts (α‐SMA^?, CAV⁺, FAP⁺, VIM⁺) into CAF‐like fibroblasts (α‐SMA⁺, CAV^?, FAP⁺, VIM⁺). Fibroblasts activated by early‐stage cancer‐exosomes (SW480‐Exos) are highly pro‐proliferative and pro‐angiogenic and display elevated expression of pro‐angiogenic (IL8, RAB10, NDRG1) and pro‐proliferative (SA1008, FFPS) proteins. In contrast, fibroblasts activated by late‐stage cancer‐exosomes (SW620‐Exos) display a striking ability to invade through extracellular matrix through upregulation of pro‐invasive regulators of membrane protrusion (PDLIM1, MYO1B) and matrix‐remodeling proteins (MMP11, EMMPRIN, ADAM10). Conserved features of Exos‐mediated fibroblast activation include enhanced ECM secretion (COL1A1, Tenascin‐C/X), oncogenic transformation, and metabolic reprogramming (downregulation of CAV‐1, upregulation of glycogen metabolism (GAA), amino acid biosynthesis (SHMT2, IDH2) and membrane transporters of glucose (GLUT1), lactate (MCT4), and amino acids (SLC1A5/3A5)). This study highlights the role of primary and metastatic CRC tumor‐derived Exos in generating phenotypically and functionally distinct subsets of CAFs that may facilitate tumor progression.     

The Embryos Obtained from the Karyoplasts of Starfish Oocytes: Their Development and Sensitivity to Cytostatic Neurotransmitter Antagonists

Karyoplasts obtained from full-grown oocytes of the starfish Aphelasterias japonica have practically no cytoplams and are incapable of maturation. Karyoplasts of oocytes of starfishes Marthasterias glacialis and Acanthaster planci have the cytoplasm (10%–15% of the total karyoplast volume) and are often capable of maturation, fertilization and one or several cleavage divisions. The embryoskaryoplasts completely lose supersensitivity and retain usual sensitivity to cytostatic antagonists of neurotransmitters. The assumption is made that the incapability or limited capability of this embryos for development might be due to a deficiency of certain components of the "prenervous" neurotransmitter systems.     

Different Oxidative Stress Response in Keratinocytes and Fibroblasts of Reconstructed Skin Exposed to Non Extreme Daily-Ultraviolet Radiation

Experiments characterizing the biological effects of sun exposure have usually involved solar simulators. However, they addressed the worst case scenario i.e. zenithal sun, rarely found in common outdoor activities. A non-extreme ultraviolet radiation (UV) spectrum referred as “daily UV radiation” (DUVR) with a higher UVA (320–400 nm) to UVB (280–320 nm) irradiance ratio has therefore been defined. In this study, the biological impact of an acute exposure to low physiological doses of DUVR (corresponding to 10 and 20% of the dose received per day in Paris mid-April) on a 3 dimensional reconstructed skin model, was analysed. In such conditions, epidermal and dermal morphological alterations could only be detected after the highest dose of DUVR. We then focused on oxidative stress response induced by DUVR, by analyzing the modulation of mRNA level of 24 markers in parallel in fibroblasts and keratinocytes. DUVR significantly modulated mRNA levels of these markers in both cell types. A cell type differential response was noticed: it was faster in fibroblasts, with a majority of inductions and high levels of modulation in contrast to keratinocyte response. Our results thus revealed a higher sensitivity in response to oxidative stress of dermal fibroblasts although located deeper in the skin, giving new insights into the skin biological events occurring in everyday UV exposure.     

Ultrastructure of Bacteriomes and Their Sensitivity to Ambient Temperatures in Prostephanus truncatus (Horn)

In light microscopical sections of Prostephanus truncatus (Horn) (Col.: Bostrychidae) it was shown that both larvae and adults have a pair of bacteriomes dorsally located in the fat body parallel to the midgut. Bacteriome development was shown mainly to occur during larval stages. Bacteriome size was not found to be associated with body size in adults, but in larvae reared at 30°C bacteriome size increased progressively with body length. The shape of the bacteriomes varied from round to conic-oval, but a common feature was that they were larger and rounded in older larvae and females as compared to males, where they usually appeared more shrunken and slightly deformed. Electron microscopy of thin sections showed that the bacteriomes were composed of multinucleate syncytia surrounded by a layer of boundary cells. The syncytia harboured many small coccoid bacteroids. Typical eukaryotic organelles were found in the cytoplasm of the bacteriomes. These and other structural features were outlined. The effect of rearing temperatures at 30, 35 and 37°C on bacteriome development in larvae and adults was examined. The symbiotes could not be eliminated but a significant reduction of bacteriome size was found in females reared at 35°C and 37°C as compared to specimens grown at 30°C. A possible association of bacteriome size and reproduction was evaluated by transferring P. truncatus specimens reared at 35°C and 37°C to 30°C for two months and counting the number of offspring; their reproduction was compared with controls kept at 30°C throughout the experiment. Specimens from 35°C and 37°C had significantly lower reproduction rates than controls. The potential implications of heat sensitivity of bacteriomes of P.truncatus is discussed in an integrated pest management context.     

Synaptic and Non-Synaptic Mitochondria in Hippocampus of Adult Rats Differ in Their Sensitivity to Hypothyroidism

Hypothyroidism in humans provokes various neuropsychiatric disorders, movement, and cognitive abnormalities that may greatly depend on the mitochondrial energy metabolism. Brain cells contain at least two major populations of mitochondria that include the non-synaptic mitochondria, which originate from neuronal and glial cell bodies (CM), and the synaptic (SM) mitochondria, which primarily originate from the nerve terminals. Several parameters of oxidative stress and other parameters in SM and CM fractions of hippocampus of adult rats were compared among euthyroid (control), hypothyroid (methimazol-treated), and thyroxine (T4)-treated hypothyroid states. nNOS translocation to CM was observed with concomitant increase of mtNOS??s activity in hypothyroid rats. In parallel, oxidation of cytochrome c oxidase and production of peroxides with substrates of complex I (glutamate?+?malate) were enhanced in CM, whereas the activity of aconitase and mitochondrial membrane potential (????_m) were decreased. Furthermore, the elevation of mitochondrial hexokinase activity in CM was also found. No differences in these parameters between control and hypothyroid animals were observed in SM. However, in contrast to CM, hypothyroidism increases the level of pro-apoptotic K-Ras and Bad in SM. Our results suggest that hypothyroidism induces moderate and reversible oxidative/nitrosative stress in hippocampal CM, leading to the compensatory elevation of hexokinase activity and aerobic glycolysis. Such adaptive activation in glycolytic metabolism does not occur in SM, suggesting that synaptic mitochondria differ in their sensitivity to the energetic disturbance in hypothyroid conditions.     

Different Growth Responses of Hamster Dermal Fibroblasts and Chondrocytes to Platelet–derived Growth Facter in Cell Culture

The growth responses of dermal fibroblasts and chondrocytes obtained from 1-week-old hamsters to a growth factor from platelets [platelet-derived growth factor (PDGF)] were compared. Autoradiography showed that most of the whole nuclei of dermal fibroblasts were labeled with ³H-thymidine in 5% whole blood serum (WBS), which contains platelet releasate, but only a few percent were labeled in 5% plasma-derived serum (PDS), which is free of platelet releasate. Addition of platelet releasate to PDS restored the growth stimulatory activity of the serum for dermal fibroblasts. In contrast, most of the nuclei of chondrocytes were labeled with ³H-thymidine both in 5% WBS and 5% PDS. Dermal fibroblasts grew in 5% WBS, but not in 5% PDS, whereas chondrocytes grew in both 5% WBS and 5% PDS. Partially purified PDGF added to the medium with 0.5% fetal calf serum stimulated DNA synthesis in dermal fibroblasts, but not in chondrocytes.     

Sensitivity to and Degradation of Pentachlorophenol by Phanerochaete spp

This research measured mycelial extension rates of selected strains of Phanerochaete chrysorhiza, Phanerochaete laevis, Phanerochaete sanguinea, Phanerochaete filamentosa, Phanerochaete sordida, Inonotus circinatus, and Phanerochaete chrysosporium and the ability of these organisms to tolerate and degrade the wood preservative pentachlorophenol (PCP) in an aqueous medium and in soil. Most of the tested species had mycelial extension rates in the range of ≤0.5 to 1.5 cm day⁻¹, but there were large interspecific differences. A notable exception, P. sordida, grew very rapidly, with an average mycelial extension rate of 2.68 cm day⁻¹ at 28°C. Rank of species by growth rate was as follows: P. chrysosporium > P. sordida > P. laevis > P. chrysorhiza = P. sanguinea > I. circinatus = P. filamentosa. There were also significant intraspecific differences in mycelial extension rates. For example, mycelial extension rates among strains of P. sordida ranged from 1.78 to 4.81 cm day⁻¹. Phanerochaete spp. were very sensitive to PCP. Growth of several species was prevented by the presence of 5 ppm (5 μg/g) PCP. However, P. chrysosporium and P. sordida grew at 25 ppm PCP, albeit at greatly decreased mycelial extension rates. In an aqueous medium, mineralization of PCP by P. sordida 13 (ca. 12% after 30 days) was significantly greater than that by all other tested P. sordida strains and P. chrysosporium. After 64 days, the level of PCP had decreased by 96 and 82% in soil inoculated with P. chrysosporium and P. sordida, respectively. Depletion of PCP by these fungi occurred in a two-stage process. The first stage was characterized by a rapid depletion of PCP that coincided with an accumulation of pentachloroanisole (PCA). At the end of the first stage, ca. 64 and 71% of the PCP was converted to PCA in P. chrysosporium and P. sordida cultures, respectively. In the second stage, levels of PCP and PCA were reduced by 9.6 and 18%, respectively, in soil inoculated with P. chrysosporium and by 3 and 23%, respectively, in soil inoculated with P. sordida. PCA was mineralized by both P. chrysosporium and P. sordida in an aqueous medium.     

Identification of Fungal Species Associated with Cladode Spot of Prickly Pear and Their Sensitivity to Chitosan

México is the most important producer of prickly pear (Opuntia ficus‐indica) in the world. There are several fungal diseases that can have a negative impact on their yields. In this study, there was a widespread fungal richness on cladodes spot of prickly pears from México. A total of 41 fungi isolates were obtained from cladodes spot; 11 of them were morphologically different. According to the pathogenicity test, seven isolates caused lesions on cladodes. The morphological and molecular identification evidenced the isolation of Colletotrichum gloeosporioides, Alternaria alternata, Fusarium lunatum, Curvularia lunata. All these species caused similar symptoms of circular cladodes spot. However, it is noticeable that some lesions showed perforation and detachment of affected tissues by Fusarium lunatum. To our knowledge, this is the first report of the Fusarium lunatum as phytopathogenic fungus of cladodes of prickly pear. The chitosan inhibited the mycelium growth in the seven isolates of phytopathogenic fungi. Chitosan applications diminished the disease incidence caused by C. gloeosporioies and F. lunatum in 40 and 100%, respectively. Likewise, the lesion severity index in cladodes decreased. There are no previous reports about the application of chitosan on cladodes of prickly pears for the control of phytopathogenic fungi. Therefore, this research could contribute to improve the strategies for the management of diseases in prickly pear.     

不同溶剂分级提取的茶多糖的组成及降血糖活性

比较绿茶中不同溶剂分级提取的茶多糖的得率、含量、单糖组成及降血糖活性的差异,以从茶叶中分离出高活性的茶多糖.粗老绿茶采用去离子水低温提取茶叶中的水溶性多糖TPSⅠ,水不溶性多糖先用草酸铵提取其果胶类多糖TPSⅡ,再用碱提取,所得到碱提取液回调至pH中性后可溶解部分为碱溶性多糖TPSⅢ,并分析其得率、含量、采用气相色谱和离子色谱分析了其单糖组成,通过四氧嘧啶腹腔注射造成高血糖模型,连续灌胃茶多糖12 d,对比研究两种茶多糖的降血糖活性.并比较复合纤维素酶提取与去离子水提取在得率、含量、单糖组成及其降血糖活性的差异.TPSⅠ单糖组成以鼠李糖、半乳糖和半乳糖醛酸为主;TPSⅡ以半乳糖和半乳糖醛酸为主;TPSⅢ以阿拉伯糖、葡萄糖和半乳糖为主.降血糖活性顺序为:TPSⅠ＞TPSⅢ＞TPSⅡ,但TPSⅠ得率最低.复合纤维素酶提取的多糖得率、糖醛酸提取率比热水提取的多糖高1.53倍和1.42倍.酶提茶多糖较水提茶多糖降血糖效果更明显些.表明茶叶中水溶性多糖降血糖活性最好,复合纤维素酶提取可以提高其得率,增强其活性.     

Compensation and resistance to herbivory in seagrasses: induced responses to simulated consumption by fish

Herbivory can induce changes in plant traits that may involve both tolerance mechanisms that compensate for biomass loss and resistance traits that reduce herbivore preference. Seagrasses are marine vascular plants that possess many attributes that may favour tolerance and compensatory growth, and they are also defended with mechanisms of resistance such as toughness and secondary metabolites. We quantified phenotypic changes induced by herbivore damage on the temperate seagrass Posidonia oceanica in order to identify specific compensatory and resistance mechanisms in this plant, and to assess any potential trade-offs between these two strategies of defence. We simulated three natural levels of fish herbivory by repeatedly clipping seagrass leaves during the summer period of maximum herbivory. Compensatory responses were determined by measuring shoot-specific growth, photosynthetic rate, and the concentration of nitrogen and carbon resources in leaves and rhizomes. Induced resistance was determined by measuring the concentration of phenolic secondary metabolites and by assessing the long-term effects of continued clipping on herbivore feeding preferences using bioassays. Plants showed a significant ability to compensate for low and moderate losses of leaf biomass by increasing aboveground growth of damaged shoots, but this was not supported by an increase in photosynthetic capacity. Low levels of herbivory induced compensatory growth without any measurable effects on stored resources. In contrast, nitrogen reserves in the rhizomes played a crucial role in the plant’s ability to compensate and survive herbivore damage under moderate and high levels of herbivory, respectively. We found no evidence of inducibility of long-term resistance traits in response to herbivory. The concentration of phenolics decreased with increasing compensatory growth despite all treatments having similar carbon leaf content, suggesting reallocation of these compounds towards primary functions such as cell-wall construction.     

人食管平滑肌条在不同pH值溶液中收缩及舒张反应研究

目的:观察不同pH值的Kreb's液对人食管下段平滑肌条的体外收缩功能的影响,同时明确食管环形肌(circular smoothmuscle,CM)及食管纵行肌(longitude smooth muscle,LM)对其反应的区别,以探讨酸性反流性食管炎及Barrett食管的相关发病机制.方法:根据24h食管pH值检测、食管镜检及HE染色结果,分辨、筛选正常人食管平滑肌,制备环形平滑肌与纵行平滑肌条,描述4种pH值Kreb's液(pH∶6.7,6.9,7.1,7.3)组给药前后人食管肌条收缩曲线.结果:LM在各组pH条件下乙酰胆碱的浓度依赖收缩反应无明显差异,但是CM在pH为6.7组中收缩反应明显低于其他各组;LM在各pH组中对异丙肾上腺素舒张反应无明显差异,而CM在pH为6.7及6.9组异丙肾上腺素舒张反应明显降低.结论:不同的酸性条件下食管平滑肌中的纵行肌及环行肌对乙酰胆碱及异丙肾上腺素的反应性不同,酸损伤减弱CM收缩及舒张反应,可能参与反流性食管炎、Barrett食管等食管运动功能性疾病的发生过程.     

Biochemical Basis of Different Sensitivity to Methotrexate in Daucus carota and Oryza sativa Cell Cultures

The biochemical basis of the different sensitivity to methotrexateof Daucus carota and Oryza sativa cell cultures has been investigated.Carrot cells have a dihydrofolate reductase activity about tentimes higher than rice cells. In addition, they show a loweruptake rate of the inhibitor. No relevant differences have beenfound in the K_m value for the dihydrofolate of the two enzymesand in the degree of inhibition of their activity by methotrexate. Key words: Dihydrofolate reductase, Methotrexate resistance, Plant cell suspension cultures, Oryza sativa, Daucus carota     

不同粒级土壤团聚体呼吸特征及其对碳排放的贡献

通过土盆培养试验研究了不同粒级黄棕壤团聚体呼吸特征及其对碳排放的贡献,结果表明:各粒级土壤团聚体及原状土在培养初期土壤呼吸速率较高,之后逐渐降低。在整个培养期间原状土保持了最大的土壤呼吸速率;土壤团聚体类型对土壤呼吸速率影响较大,3种粒级团聚体土壤呼吸速率大小表现为(5 mm)(1 mm)≈(1~5 mm),其中以5 mm团聚体对土壤碳排放的贡献最大。相关性分析表明,指数模型能较好地描述不同粒级团聚体的土壤呼吸对温度变化的响应;Q10值介于2.53~5.11之间且与土壤有机碳、全氮含量的变化规律基本一致,表现为1 mm团聚体有机碳、全氮含量和Q10值较大,5 mm、1~5 mm团聚体有机碳、全氮含量和Q10值相对较低,说明土壤团聚体粒级越小,其呼吸速率对温度越敏感。土壤有机碳、氮含量与土壤结构是影响土壤呼吸温度敏感性的重要因素。