Histochemistry

组织化学
  • 文章类型: Journal Article
    ArgyreiaLour属中的三种牵牛花物种。,A.lycioides(Choisy)Traiperm&Rattanakrajang,A.mekongensisGagnep&Courchet,和A.versicolor(Kerr)Staples&Traiperm,被发现是同时发生和共同开花的。Argyreiamekongensis和A.versicolor是罕见的,而A.lycioides几乎受到威胁,分布在缅甸和泰国各地。我们调查了关键的花卉特征(花卉形态和物候,以及花蜜腺盘和雄蕊毛状体的微观形态),并筛选了推测有助于传粉媒介吸引的重要化合物。我们的发现表明,花卉形态的某些方面(例如,花冠尺寸,肢体存在,和花色)的三个研究同源物表现出显着差异。此外,传粉者的组成似乎受花的形状和大小的影响;花冠管较宽的牵牛花物种被较大的蜜蜂授粉。所有物种的花蜜腺盘的形态相似,而在不同物种中观察到雄蕊毛状体的变化。在所有三个物种中都发现了腺毛,而非腺毛体仅在A.versicolor中发现。组织化学结果表明,每个物种的花蜜腺和雄蕊毛状体中存在不同的化合物,这可能有助于花卉吸引力和防御。这些发现表明,在同伴共同开花的牵牛花物种中,花卉游客之间存在一些隔离,这似乎受到花及其化合物的宏观和微观形态的影响。此外,了解这些同胞共同开花的Argyreia物种的花卉形态和化学引诱剂可能有助于维持其常见的传粉者,以保护这些稀有和濒危物种,尤其是A.versicolor.
    Three morning glory species in the genus Argyreia Lour., A. lycioides (Choisy) Traiperm & Rattanakrajang, A. mekongensis Gagnep & Courchet, and A. versicolor (Kerr) Staples & Traiperm, were found co-occurring and co-flowering. Argyreia mekongensis and A. versicolor are rare, while A. lycioides is near threatened and distributed throughout Myanmar and Thailand. We investigated key floral characters (floral morphology and phenology, as well as the micromorphology of the floral nectary disc and staminal trichomes) and screened for important chemical compounds hypothesized to contribute to pollinator attraction. Our findings demonstrate that some aspects of floral morphology (e.g., corolla size, limb presence, and floral color) of the three studied congeners exhibit significant differences. Moreover, pollinator composition appears to be influenced by floral shape and size; morning glory species with wider corolla tubes were pollinated by larger bees. The morphology of the floral nectary disc was similar in all species, while variation in staminal trichomes was observed across species. Glandular trichomes were found in all three species, while non-glandular trichomes were found only in A. versicolor. Histochemical results revealed different compounds in the floral nectary and staminal trichomes of each species, which may contribute to both floral attraction and defense. These findings demonstrate some segregation of floral visitors among sympatric co-flowering morning glory species, which appears to be influenced by the macro- and micromorphology of flowers and their chemical compounds. Moreover, understanding the floral morphology and chemical attractants of these sympatric co-flowering Argyreia species may help to maintain their common pollinators in order to conserve these rare and endangered species, especially A. versicolor.
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  • 文章类型: Journal Article
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  • 文章类型: Journal Article
    煤烟霉菌是腐生附生真菌,主要生长在昆虫分泌物上,但它们也可能与植物分泌物有关。在这项研究中,我们的目的是描述阿非纳西木烟霉菌与Azadirachtaindica的外芽蜜腺的相互作用。在没有和有C.alfenasii侵染的in草外芽蜜腺的连续切片上进行了解剖学和组织化学研究。测定了分泌花蜜的可溶性总糖含量,并评估了真菌在蒸馏水以及葡萄糖和花蜜溶液中的分生孢子萌发。A的芽蜜腺是细长的结构,成对出现在叶柄基部附近。渗出的花蜜每µL花蜜平均含有534.8µg的总可溶性糖,为分生孢子萌发和真菌生长提供了理想的条件。C.alfenasii菌丝在蜜腺上生长,穿透角质层的裂缝,在角质层下行进,并通过细胞间和细胞内途径穿透分泌组织。本报告是第一个描述C.alfenasii与A.indica蜜腺的相互作用的报告,包括菌丝渗透到蜜腺组织和植物防御机制。
    Sooty moulds are saprophytic epiphytic fungi that grow mostly on insect secretions, but they can also be associated with plant secretions. In this study, we aimed to describe de interaction of Capnodium alfenasii sooty mould with the extrafloral shoot nectaries of Azadirachta indica. Anatomical and histochemical studies were carried out on serial sections of extrafloral shoot nectaries of A. indica without and with C. alfenasii infestation. The total soluble sugar content of the secreted nectar was determined, and the conidial germination of the fungus in distilled water and in dextrose and nectar solutions was evaluated. The shoot nectaries of A. indica are elongated structures that occur in pairs near the base of the petiole. The exuded nectar contains an average of 534.8 µg of total soluble sugars per µL of nectar and provides ideal conditions for conidial germination and fungal growth. C. alfenasii hyphae grow on the nectary, penetrate through breaks in the cuticle, travel under the cuticle and penetrate the secretory tissue by inter- and intracellular routes. The present report is the first to describe the interaction of C. alfenasii with the A. indica nectary, including the penetration of hyphae into nectariferous tissues and the plant defence mechanisms.
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  • 文章类型: Journal Article
    胆虫寄主Eugeniauniflora(Myrtaceae)适应不同的光照条件,使叶片能够在阳光和阴凉处生产和生存。阴暗环境中的E.uniflora的叶子有更多的叶肉层,Clinodiplosisprofusa(Cecidomyiidae)的gall更大,更宽。基于这些先前的观察,这项研究调查了C.profusa在不同光照条件下在E.uniflora叶片上诱导的galls的形态发生,揭示虫子是否有适应的潜力,用树叶观察到的。为此,我们比较了解剖学,组织学,以及在阳光和阴凉环境下不同发育阶段叶片和gall的组织化学发育。此外,我们分析了构成成熟胆壁的组织的细胞学特征。遮荫的细胞在发育阶段结束时扩张得更多,这可能解释了在先前的研究中发现的更大体积的遮阳。然而,在成熟阶段,在对比的光照条件下,这些gall在组织厚度和最终细胞伸长方面没有显着差异。在超微结构分析中,成熟的gall显示出区分内外薄壁组织细胞的梯度。内部薄壁组织有营养细胞,具有致密的细胞质和丰富的细胞器。营养细胞中淀粉颗粒的积累更高,在最内层检测到的淀粉颗粒水解的证据导致还原糖的积累,which,有了质体和蛋白体,是与胆汁诱导剂接触的细胞中氧化应激消散的重要机制。
    The gall-host Eugenia uniflora (Myrtaceae) is adaptable to different light conditions, enabling leaf production and survival in both sun and shade. Leaves of E. uniflora in shaded environments have more mesophyll layers, and galls of Clinodiplosis profusa (Cecidomyiidae) are larger and wider. Based on these previous observations, this study investigated the morphogenesis of galls induced by C. profusa on leaves of E. uniflora in different light conditions, revealing if the galls have a potential for acclimation, as observed with leaves. For this purpose, we compared the anatomical, histometric, and histochemical development of leaves and galls at different stages of development in sun and shade environments. Additionally, we analyzed the cytological features of the tissues composing the mature gall walls. Cells of shade galls expanded more toward the end of the developmental phase, which may explain the larger volume found for shade galls in a previous study. However, during the mature phase, these galls showed no significant differences in tissue thickness and final cell elongation in the contrasting light conditions. In the ultrastructural analyses, mature galls showed a gradient distinguishing the outer and inner parenchyma cells. The inner parenchyma had nutritive cells, with dense cytoplasm and abundant organelles. A higher accumulation of starch grains in nutritive cells, with evidence of hydrolysis of starch grains detected in the innermost layers leads to the accumulation of reducing sugars, which, with the presence of plastoglobules and protein bodies, are important mechanisms of oxidative stress dissipation in the cells in contact with the gall inducer.
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  • 文章类型: Journal Article
    十字花科,通常被称为十字花科植物,在全球范围内种植和消费,芸苔属以其功能成分而闻名。这些蔬菜富含营养和促进健康的植物化学物质,近年来受到越来越多的关注。这项研究提出了一个全面的微观,色谱,哈萨克斯坦甘蓝型油菜种子的光谱表征旨在阐明其形态特征和化学成分。显微分析显示黄酮类化合物的不同定位,总脂质,和生物碱。种子提取物的高效薄层色谱(HPTLC)分析显示出复杂的化学特征,己烷提取物中存在大量非极性化合物。此外,甲醇提取物揭示了多种化合物的存在,包括生物碱,黄酮类化合物,和芥子油苷.化学成分在不同的芸苔属物种之间表现出品种差异,与B.napusL.种子显示更高浓度的生物活性化合物。此外,液相色谱-四极杆飞行时间质谱(LC-QToF-MS)分析提供了对化学成分的见解,与芥子碱异构体,阿魏酸,和芥子酰胆碱衍生物作为种子中的主要化合物。本研究有助于更好地理解欧洲油菜种子的化学多样性和质量控制方法,强调它们在功能性食品和营养品应用中的重要性。
    The Brassicaceae family, commonly referred to as cruciferous plants, is globally cultivated and consumed, with the Brassica genus being particularly renowned for its functional components. These vegetables are rich sources of nutrients and health-promoting phytochemicals, garnering increased attention in recent years. This study presents a comprehensive microscopic, chromatographic, and spectroscopic characterization of Brassica napus L. seeds from Kazakhstan aimed at elucidating their morphological features and chemical composition. Microscopic analysis revealed distinct localization of flavonoids, total lipids, and alkaloids. High-performance thin-layer chromatography (HPTLC) analysis of seed extracts demonstrated a complex chemical profile with significant quantities of non-polar compounds in the hexane extracts. Additionally, methanolic extracts revealed the presence of diverse chemical compounds, including alkaloids, flavonoids, and glucosinolates. The chemical composition exhibited varietal differences across different Brassica species, with B. napus L. seeds showing higher concentrations of bioactive compounds. Furthermore, liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QToF-MS) analysis provided insights into the chemical composition, with sinapine isomers, feruloyl, and sinapoyl choline derivatives as major compounds in the seeds. This study contributes to a better understanding of the chemical diversity and quality control methods\' approximations of B. napus L. seeds, highlighting their importance in functional food and nutraceutical applications.
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  • 文章类型: Journal Article
    原位RT-PCR由于其快速加工和低成本设备而具有优于其他表达分析方法的优势。然而,这项技术并非没有挑战。提出了一种基于由离心管制成的胶囊的协议,该协议具有优于载玻片的优势。这种胶囊保护组织学切片不干燥,和它的容易组装减少了孵化之间的暂停时间。此外,放置样品的容器尺寸允许添加和取出不同的溶液。每次孵育后,胶囊不需要事先密封,and,最重要的是,它是一种低成本和易于获得的材料。还描述了使用低温恒温器进行组织切片的指南,该指南提供了优于其他切片方法的优势。
    In situ RT-PCR presents advantages over other expression analysis methods due to its rapid processing and low-cost equipment. However, this technique is not without its challenges. A protocol based on a capsule made from centrifuge tubes that offers advantages over slides is presented. This capsule protects histological sections from drying out, and its easy assembly reduces time pauses between incubations. In addition, the container size where the sample is deposited allows the addition and withdrawal of the different solutions. The capsule does not need previous sealing after each incubation, and, above all, it is a low-cost and accessible material. A guideline for tissue sectioning using a cryostat that offers advantages over other sectioning methods is also described.
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  • 文章类型: Journal Article
    甲型流感病毒的组织嗜性和广泛宿主范围由唾液酸(SA)α2,3-Gal和SAα2,6-Gal受体的存在决定。最近的研究表明,具有两种受体的动物允许重排和出现具有公共卫生重要性的新病毒株。本研究旨在使用凝集素免疫组织化学方法评估9种新热带蛇种中人和禽流感A受体的表达和分布。我们选择了2019年至2023年在米纳斯吉拉斯州联邦大学兽医病理学部门进行尸检的17条蛇。鼻甲切片,气管,肺,口腔粘膜,使用凝集素进行胃和肠的免疫组织化学分析。这项研究发现,第一次,SAα2,3-Gal和SAα2,6-Gal受体在蛇的呼吸道和消化道中的共表达,表明这些物种可能对禽类和人类来源的甲型流感病毒敏感。因此,蛇可以被认为是监测野生甲型流感的重要物种,城市和城郊环境。应该进行更多的研究来调查蛇在甲型流感流行病学中的作用。
    The tissue tropism and the wide host range of influenza A viruses are determined by the presence of sialic acid (SA) α2,3-Gal and SA α2,6-Gal receptors. Recent studies have shown that animals possessing both receptors allow for the rearrangement and emergence of new viral strains of public health importance. This study aimed to evaluate the expression and distribution of human and avian influenza A receptors in nine Neotropical snake species using lectin immunohistochemistry. We selected 17 snakes that were examined post mortem at the Veterinary Pathology Sector of the Universidade Federal de Minas Gerais between 2019 and 2023. Sections of nasal turbinate, trachea, lung, oral mucosa, stomach and intestine were subjected to immunohistochemical analysis using the lectins Maackia amurensis and Sambucus nigra. This research detected, for the first time, co-expression of SA α2,3-Gal and SA α2,6-Gal receptors in the respiratory and digestive tracts of snakes, indicating the possible susceptibility of these species to influenza A virus of avian and human origin. Consequently, snakes can be considered important species for monitoring influenza A in wild, urban and peri-urban environments. More studies should be conducted to investigate the role of snakes in influenza A epidemiology.
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  • 文章类型: Journal Article
    聚苯乙烯纳米塑料和二氧化钛纳米颗粒在所有环境中广泛传播,通常在相同的框架内共存。这两种污染物都会对细胞和植物生理产生负面影响,关注它们之间可能的相互作用,这可能会增加彼此对植物的有害影响。尽管这个问题很紧迫,很少有文献解决这个问题。为了评估这种共同污染的潜在风险,扁豆种子用聚苯乙烯纳米塑料和二氧化钛纳米颗粒(锐钛矿晶型)处理五天,单独和共同存在。细胞学分析,在分离的芽和根上进行了氧化应激的组织化学和生化评估。TEM分析似乎表明两种纳米材料之间不存在物理/化学相互作用。同种关系下的幼苗显示出最大的细胞毒性和遗传毒性作用,以及与生长抑制相关的高水平的氧化应激标记物。即使生化数据没有证据表明单独用聚苯乙烯纳米塑料处理或与二氧化钛纳米颗粒共存的材料之间存在显着差异,组织化学分析强调了氧化标记的不同模式,表明两种纳米材料的协同作用。InAccording,在共处理下,根和芽中与纳米塑料相关的荧光信号更高,也许是由于众所周知的二氧化钛纳米粒子诱导根组织损伤的能力,以这种方式促进聚苯乙烯纳米塑料的吸收和转运到植物体中。在抗氧化机械中,过氧化物酶活性显示处理的根显著增加,特别是在共同处理下,可能与应激诱导的木质素合成有关,而不是与过氧化氢解毒有关。目前的结果清楚地表明,金属纳米颗粒对纳米塑料对植物的负面影响正在恶化,强调了考虑到不同纳米材料共同处理影响的研究的重要性,可以更好地反映复杂的环境条件。
    Polystyrene nanoplastics and titanium dioxide nanoparticles are widely spread in all environments, often coexisting within identical frameworks. Both these contaminants can induce negative effects on cell and plant physiology, giving concerns on their possible interaction which could increase each other\'s harmful effects on plants. Despite the urgency of this issue, there is very little literature addressing it. To evaluate the potential risk of this co-contamination, lentil seeds were treated for five days with polystyrene nanoplastics and titanium dioxide nanoparticles (anatase crystalline form), alone and in co-presence. Cytological analyses, and histochemical and biochemical evaluation of oxidative stress were carried out on isolated shoots and roots. TEM analysis seemed to indicate the absence of physical/chemical interactions between the two nanomaterials. Seedlings under cotreatment showed the greatest cytotoxic and genotoxic effects and high levels of oxidative stress markers associated with growth inhibition. Even if biochemical data did not evidence significant differences between materials treated with polystyrene nanoplastics alone or in co-presence with titanium dioxide nanoparticles, histochemical analysis highlighted a different pattern of oxidative markers, suggesting a synergistic effect by the two nanomaterials. In accordance, the fluorescence signal linked to nanoplastics in root and shoot was higher under cotreatment, perhaps due to the well-known ability of titanium dioxide nanoparticles to induce root tissue damage, in this way facilitating the uptake and translocation of polystyrene nanoplastics into the plant body. In the antioxidant machinery, peroxidase activity showed a significant increase in treated roots, in particular under cotreatment, probably more associated with stress-induced lignin synthesis than with hydrogen peroxide detoxification. Present results clearly indicate the worsening by metal nanoparticles of the negative effects of nanoplastics on plants, underlining the importance of research considering the impact of cotreatments with different nanomaterials, which may better reflect the complex environmental conditions.
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  • 文章类型: Journal Article
    对家猪(Susscrofadomesticus)的角膜进行了全面的光和超微结构检查,发现了四个不同的层:前上皮,角膜基质,Descemet膜和内皮。虽然鲍曼的层没有通过组织学明确识别,组织化学分析表明存在一个基本的鲍曼层,可能是进化造成的。角膜外表面的扫描电子显微镜揭示了两种细胞类型,以微投影为特征,光细胞表现得更短,与暗细胞相比,投影更厚。通过扫描电子显微镜检查内表面,显示内皮层没有纤毛和微绒毛,然而,观察到微弱的圆形到椭圆形的高度,可能代表细胞核。透射电镜显示,前上皮的基底细胞与基底膜紧密粘附,沿着基面具有半桥粒。这些基底细胞通过指间和一些桥粒广泛地相互连接。表面细胞层由几排紧密附着的扁平细胞组成,形成带有透明带闭塞的防漏层。该层的最外面的细胞显示出精细的突起以增强表面积,促进泪膜分布。在较低的放大倍数下,角膜基质的透射电子显微镜显示明带和暗带交替,浅色带代表胶原蛋白原纤维薄片的横截面,深色带对应于纵向或倾斜部分。梭形角膜细胞(成纤维细胞)被鉴定为原代基质细胞,交织在薄片之间,并具有与相邻角膜细胞紧密接触的长过程。总的来说,猪角膜的组织形态类似于人类角膜,除了鲍曼膜不明显。这种对猪正常角膜结构的详细了解对生物医学研究具有重要意义。为涉及该动物模型的研究提供了有价值的参考。
    A comprehensive light and ultrastructural examination of the cornea in Domestic Pigs (Sus scrofa domesticus) revealed four distinct layers: the anterior epithelium, corneal stroma, Descemet\'s membrane and endothelium. Although Bowman\'s layer was not distinctly identified through histology, histochemical analysis indicated the presence of a rudimentary Bowman\'s layer, possibly vestigial from evolution. Scanning electron microscopy of the outer corneal surface unveiled two cell types, characterized by micro-projections, with light cells exhibiting shorter, thicker projections compared to dark cells. Examination of the inner surface via scanning electron microscopy demonstrated an endothelial layer devoid of cilia and microvilli, yet faint round to oval elevations were observed, potentially representing cell nuclei. Transmission electron microscopy unveiled that basal cells of the anterior epithelium closely adhered to the basement membrane, featuring half desmosomes along the basal surface. These basal cells extensively interconnected through interdigitations and a few desmosomes. The superficial cell layer consisted of a few rows of closely attached flat cells, forming a leak-proof layer with zona occludens. The outermost cells of this layer displayed fine projections to enhance the surface area, facilitating tear film distribution. At lower magnification, Transmission electron microscopy of the corneal stroma revealed alternating light and dark bands, with light bands representing transverse sections of collagen fibril lamellae and dark bands corresponding to longitudinal or oblique sections. Spindle-shaped keratocytes (fibroblasts) were identified as the primary stromal cells, intermingled between the lamellae, and featured long processes in close contact with neighbouring keratocytes. Overall, the histomorphology of the pig cornea resembles that of the human cornea except indistinct Bowman\'s membrane. This detailed understanding of the normal corneal structure in pigs hold great significance for biomedical research, providing a valuable reference for studies involving this animal model.
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  • 文章类型: Journal Article
    这项研究的目的是描述霍加皮舌的形态,并将结果与包括浏览器在内的其他反刍动物进行比较,中间体和食草动物。该材料是在动物园中的两只动物死后收集的。霍加皮舌的结构,聚焦舌头的形状,舌面,它的乳头和舌腺,使用总体形态学检查,光和偏振显微镜,和扫描电子显微镜。霍加皮舌的特征是在舌背(舌圆环除外)和整个腹侧表面上有深色色素沉着。观察到两种类型的丝状乳头,额外的,甚至在他们的基地有6-8个预测。圆形菌形乳头以更高的密度存在,在舌尖的腹侧区域上,最高可达16/cm2。圆形和细长的瓣膜乳头在舌体和舌根之间以两条平行线排列。在其瓣膜的上皮内检测到许多味蕾,而真菌状乳头的味蕾稀疏。注意到缺乏叶状乳头。非常小的圆锥形乳头,一些透镜状的形状,出现在语言圆环上。在舌乳头的结缔组织中,I型胶原纤维比III型胶原纤维占优势。舌腺中粘液腺泡单位占优势,表明霍加皮舌腺的分泌主要是粘液。在许多方面,霍加皮的舌头类似于其他反刍动物的舌头。特定的舌面形状和舌面,连同舌腺,支持植物食品的加工,如幼嫩柔软的叶子。尽管与其他反刍动物相比,霍加皮舌的特征是圆锥形乳头较小,它的高数量的Vallate乳头与其他浏览器中的相似,中间和放牧者。因此,味觉乳头的数量表明该特征与进食类型无关。
    The aim of this study was to describe the morphology of the tongue of the okapi, and to compare the results with other ruminants including browsers, intermediates and grazers. The material was collected post-mortem from two animals from a Zoological Garden. The structure of the okapi tongue, focusing of the shape of the tongue, lingual surface, its papillae and lingual glands, was examined using gross morphology, light and polarized microscopy, and by scanning electron microscopy. The okapi tongue was characterized by dark pigmentation on the lingual dorsum (except lingual torus) and on the whole ventral surface. Two types of filiform papillae were observed, with additional, even 6-8 projections at their base. The round fungiform papillae were present at a higher density, up to 16/cm2, on the ventro-lateral area of the lingual apex. Round and elongate vallate papillae were arranged in two parallel lines between the body and root of the tongue. Numerous taste buds were detected within the epithelium of their vallum, while fungiform papillae had sparse taste buds. A lack of foliate papillae was noted. Very small conical papillae, some lenticular in shape, were present on the lingual torus. Thick collagen type I fibers were dominant over collagen type III fibers in the connective tissue of the lingual papillae. The mucous acini units were dominant among lingual glands, indicating that the secretion of okapi lingual glands was mostly mucous. In many aspects, the tongue of okapi resembles the tongue of other ruminants. The specific lingual shape and lingual surface, together with the lingual glands, support the processing of plant food, such as young and soft leaves. Although okapi tongue is characterized by smaller conical papillae compared to other ruminants, its high number of vallate papillae is similar that found in other browsers, intermediate and grazers. Thus the number of gustatory papillae rather indicates that this feature is not related to the type of feeding.
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