In vitro root differentiation and essential-oil accumulation in Angelica archangelica

Summary We studied the relationship between root differentiation and the accumulation of essential oils in Angelica archangelica in in vitro cultures and in the intact plant. Root regeneration was obtained using stem and leaf explants subjected to treatment with the auxins indole-3-butyric acid, indole-3-acetic acid and α-naphthaleneacetic acid. In both stem and leaf explants, treatment with indole-3-butyric acid induced the highest rhizogenic response in terms of both percentage of explants with roots and number of roots per explant. Independently of hormonal treatment, stem explants produced a higher average number of roots per explant. Root meristemoids were already visible at day 7 of culture in the treatments with indole-3-butyric acid and indole-3-acetic acid; they were formed directly by cambial-cell division. In vitro-regenerated roots retained primary root structure and differentiated only two primary ephemeral ducts in the pericycle; no accumulation of essential oils was detected. Same-size roots taken from the intact plant showed secondary structure and essential-oil accumulation. The results of this study suggest that the synthesis and accumulation of essential oils in Angelica archangelica is closely linked to the differentiation of secondary secretory ducts.     

Morphometric study of human alveolar ducts based on serial sections

Interactive computerized morphometry was used for the quantitative study of the terminal airway branches (alveolar ducts) that followed the last bronchioles in three human acini. Two normal adult human lungs from the autopsy service were fixed by instillation and serial sections were prepared; three tissues blocks showing a central bronchiole were selected. Primary and secondary alveolar walls were traced and the following parameters were measured: volume, surface area (of primary and secondary septa), curvature (in equivalent radius) for branches of individual generations, and cumulative values starting with the first alveolar duct and moving peripherally. Although branching was dichotomous, we noticed considerable asymmetry in the pattern of branching and number of side branches. The branching trees of alveolar ducts that we studied comprised 6,7, and 10 generations. The average volume of ducts was 0.04-0.13 mm3, the surface area of primary walls ranged from 0.3616 to 0.7931 mm2 and of secondary septal walls from 0.0100 to 0.0647 mm2. The equivalent radius of curvature was between 22.7 and 38.1 microns. Cumulative increases of volume and surface area revealed similarity in the first five generations. Secondary walls represented only 4% (or 8% if 2 sides are considered) of the primary surface area, strengthening the view that alveoli are incompletely developed side chambers secondary to the alveolar ducts.     

珊瑚菜植株分泌道发育和分布的解剖学观察

利用植物解剖学方法对珊瑚菜（Glehnia littoralis Fr．Schmidt ex Miq．）体内分泌道的发育和分布进行了观察。结果表明,珊瑚菜的分泌道有分枝,为溶生型,由1层分泌细胞围绕腔道而成。珊瑚菜叶片的分泌道发育较早,在幼叶阶段即发育成形。在根的次生韧皮部、根状茎的皮层和靠近初生木质部的髓部、叶脉的薄壁组织、叶柄维管束周围和厚角组织内侧的薄壁组织、花序轴正对维管束的皮层薄壁组织中以及果实的果壁维管束内外侧的薄壁组织中均分布有分泌道,分泌道在珊瑚菜体内分布广泛。     

臭椿茎中分泌道的发育及其组织化学研究

利用植物解剖学方法研究臭椿茎和叶柄中分泌道的结构、分布和发育过程.结果表明:臭椿茎和叶柄中的分泌道分布于髓的周缘,次生木质部中无分泌道.分泌道是由一层分泌细胞围绕分泌腔而构成,分泌细胞外有1～2层鞘细胞.分泌道以裂生方式形成,其发育过程可分为3个阶段:原始细胞阶段、形成阶段和成熟阶段.在原始细胞阶段,一群原始细胞具浓厚细胞质,细胞核清晰可见;形成阶段,原始细胞的中央细胞间细胞壁中层降解,细胞壁分离,形成腔隙,随着分泌细胞数量的增加,分泌腔体积扩大;成熟阶段的分泌道具有12～16个分泌细胞,1～2层鞘细胞,分泌腔直径为30～50μm.组织化学研究表明,分泌细胞及分泌道内含物中含大量的萜类、多糖和脂类物质.机械创伤能够诱导次生木质部中产生创伤分泌道.臭椿茎中的分泌道和创伤性分泌道在抵御生物和非生物胁迫中起重要作用.     

Development of secretory units of mouse submandibular gland

Summary The fine structure of the secretory units of the mouse submandibular gland was studied according to the developmental sequence. The embryonic submandibular gland consists of terminal tubules and ducts. Myoepithelium is associated only with the terminal tubules, and the cells of the primary intercalated ducts show characteristics of the young striated duct cells. The major changes shortly after birth consist of: 1) opening of the secretory lumina, 2) increasing rough ER and its altered configuration, 3) dilatation of Golgi cisternae and 4) changes in the granular structure. These findings suggest that the salivary secretion first occurs after birth, and acinar differentiation or transformation of the secretory cells of the terminal tubules is induced and profoundly affected by the commencement of the secretory activity. In the intercalated ducts this process is somehow inhibited, and the granular cells found in the adult can be considered as the remnants of the secretory cells of the terminal tubules.     

Resin ducts of Pinus hatepensis Mill.–Their structure, development and pattern of arrangement

The primary resin ducts in the axis of plants of Pinus halepensis Mill, consist of two separate systems the pattern of which is correlated with the vascular systems of the organs in which they appear. These systems are: (1) ducts of the roots and the hypocotyl; (2) ducts of all the branches and juvenile leaves or scaleS. Both systems are produced by the apical meristemS. In the needles there is a third system of primary resin ducts situated in the mesophyll. These ducts are produced only to a small extent by the apical meristem of the needle and mainly by its intercalary meristem. In addition to these primary ducts of the needle, which form a separate system for each needle, at the base of the needle there may be ducts of secondary origin which are situated within the vein. These are continuous with secondary ducts of the brachyblast axis.
The secondary ducts constitute one system in the secondary xylem and phloem of the roots, branches and needle bases. They are formed by the cambium. In the xylem there are vertical and radial ducts which together form co-planar radial networkS. Each radial duct starts from a vertical duct. The first location of the stimulus for the formation of the two types of ducts is discussed. In the phloem there are only radial ducts, continuous with the radial ducts of the xylem. The cavities of the radial phloem and xylem ducts are not continuous, as there are no intercellular spaces in the region of the cambium.
The innermost vertical ducts of the secondary xylem form a kind of transitional type, in respect of their response to internal and external factors, between the primary resin ducts and the bulk of the secondary resin ducts.     

贯叶连翘的分泌结构及其与金丝桃素积累的关系

贯叶连翘（ＨｙｐｅｒｉｃｕｍｐｅｒｆｏｒａｔｕｍＬ．）地上器官分布着分泌细胞球（黑色腺点）、分泌囊（半透明腺点）和分泌道（半透明腺条）３类内部分泌结构。分泌细胞球在茎、叶和花器官中均有分布,由２层鞘细胞包围多个分泌细胞构成实心的分泌细胞团。分泌囊主要分布于叶片中,分泌道则分布于花器官中,它们都是由１～２层切向扁平细胞围绕圆形或长形腔道构成,腔道的贮存物为精油。利用组织化学方法,结合荧光显微镜观察,证实金丝桃素类物质是由分泌细胞球（黑色腺点）所合成和积累的。通过用戊二醛和锇酸固定样品的显微和超微结构观察,发现金丝桃素类物质积累在成熟腺体分泌细胞的中央大液泡中,细胞周围浓厚的细胞质中分布着大量小液泡和高尔基体、内质网等细胞器。在此基础上对金丝桃素类物质的积累过程进行了初步探讨     

Accumulation of vismione A in regenerated plants ofVismia guianensis DC

Summary The accumulation and tissue localization of antitumoral vismione A in the in vitro regenerated plants ofVismia guianensis DC. were investigated. Chemical and light and electron-microscope analyses revealed that vismione A, detected as phenolic black globules in the vacuoles, was accumulated in the leaf, mainly in the palisade, and in small amounts in the primary body of the stem (epidermis and first cortical layer). Vismione A is neither present in the secretory cavities and ducts of the leaf nor in the secretory ducts of the stem. In the leaves of the regenerated plants, the amount of vismione A reached 0.5% FW, compared to 0.1% in the leaves of the parent plant. The optimization of the in vitro regeneration of plants was obtained in MS medium enriched with BAP (1 ppm). The best results for the rooting of regenerated plants were achieved with MS medium containing half-strength salts and 10^–5 MIBA.Abbreviations APT attached proton test - BAP 6-benzyl-aminopurine - CC column chromatography - 2,4-D 2,4-dichlorophenoxyacetic acid - EtOAc ethyl acetate - IBA indole-3-butyric acid - MeOH methanol - MS Murashige and Skoog - NAA -naphthaleneacetic acid - NMR nuclear magnetic resonance - TEM transmission electron microscopy - TLC thin layer chromatography - UV ultraviolet     

The Secretory Structure of Hypericum perforatum and Its Relation to Hypericin Accumulation

Three different kinds of internal secretory structures, secretory cell globules (black dots), secretory cavities (translucent dots) and secretory ducts (translucent streaks) were observed in Hypericum perforatum L. The secretory cell globule, which occurred in flower, leaf and stem, consisted of a core of large secretory cells surrounded by two layers of flattened sheath cells. The secretory cavities were present throughout the lamina and the secretory ducts throughout the flower, both consisted of one or two layers of flattened cells walling an oil chamber or duct. Histochemical and fluorescence microscopy revealed that hypericin was accumulated in the secretory cell globules. Microscopic and ultrastmctural observation further demonstrated that hypericin was accumulated in the large central vacuole of the mature secretory cells. Numerous dictyosome, endoplasmic reticulum and small vacuoles were observed in the dense cytoplasm surrounded the large central vacuole. The process of hypericin accumulation in the secretory cells was preliminarily discussed.     

The cellular distribution of antifeedant prenylated anthranoids in the tissues ofVismia guianensis during development

Summary Accumulation and distribution of two types of phenolic antifeedant metabolites, vismiones and ferruginins, in specialized cells were investigated inVismia guianensis DC. Chemical, light, and electron-microscope analyses revealed that the two types of compounds were produced in different cells and organs and at different stages of plant development. Vismiones accumulated at various concentrations in the leaf, depending on its developmental stage, as well as in the stem of very young plants. Ferruginins appeared late in the plant's development and accumulated exclusively in secretory ducts in the secondary body of the stem and in secretory cavities of the fruits.     

Morphological and anatomical relationships of loblolly pine fine roots

 Suberized or brown roots have been traditionally considered secondary or woody tissues. The validity of using morphological features such as color to infer root anatomy for southern pines is questionable and unproven. The objectives of this study were (i) to establish relationships between root color, diameter, and developmental stage (i.e., primary or secondary tissues) for loblolly pine, (ii) to determine the percentages of primary and secondary brown roots by diameter class, and (iii) to use these percentages to make first order estimates of the amount of brown root length and surface area that is in the primary and secondary developmental stages for sampled roots of a semi-mature loblolly pine stand. ”Unsectioned” roots were collected by coring to a 25-cm depth 3 times a year and measuring roots for length and surface area by diameter class. ”Sectioned” roots were sampled from a one-time core and from periodic grab samples. These roots were sectioned and characterized by their color, diameter and developmental stage. Diameters of sectioned roots (n=353) ranged from 0.21 to 8.24 mm. White and orange roots ranged from 0.23 to 2.50 mm, while brown roots spanned the range. White roots were developmentally primary, whereas orange/brown roots were either primary (from 0.21 to 2.50 mm), secondary (from 0.33 to 8.24 mm), or in transition (from 0.27 to 0.76). Total live root length of the sampled stands was estimated to be composed of 38% primary tissue, 58% secondary tissue, and 4% transition tissue. Lastly, neither root color nor diameter was a reliable predictor of developmental stage unless roots were white (primary), or orange/brown and >2.5 mm in diameter (secondary). Received: 30 June 1997 / Accepted: 28 January 1998     

Vegetative anatomy of Pachycortnus (Anacardiaceae)

Pachycormus discolor , an arborescent desert perennial endemic to Baja California, has small, pinnately compound, hypostomatic, bifacial leaves produced on short shoots and photosynthetic stem phelloderm covered by exfoliating translucent phellem. Tightly packed laminal palisade cells are filled with tannins and lack chloroplasts. Spongy mesophyll is the major photosynthetic tissue. Leaves possess unicellular trichomes with secondary walls and uniseriate trichomes with glandular heads. Schizogenous resin ducts occur in primary phloem of stems, leaves and roots as well as all living tissues of the bark. Developmental studies reveal that initiation and differentiation of foliar primordia resembles that of other dicotyledons except that tannin cells and secretory ducts arise precociously. Primary vasculature is an open sympodial system with three principal traces diverging toward each foliar primordium. The wood is highly specialized and comprises mostly unlignified cells packed with starch grains. Thick bark is mainly produced as annual layers of secondary phloem marked by a ring of secretory ducts each surrounded by tannin cells. The possible adaptive significance of these unusual anatomical features is discussed.     

Light- and electron-microscopic radioautographic study of glycoprotein secretion in the granular duct of the submandibular gland of the male mouse

Summary L-³H-fucose was injected intravenously into adult male mice, after which, at different time intervals, the submandibular glands were removed and processed for light-and electron-microscopic radioautography. This radio active hexose was taken up by newly synthesized glycoproteins in the cells lining the granular ducts which were maximally labeled at 4 h after injection. Between 4 and 72 h the amount of labeled glycoproteins decreased moderately indicating that these macromolecules undergo a slow renewal. The main subcellular site of incorporation of 3 H-fucose into glycoproteins was the Golgi apparatus. From this organelle labeled glycoproteins were transferred to small secretory granules (diameter up to 1.0 m) located not only near the Golgi region but also throughout the apical cytoplasm. At 1 h after injection the concentration of label reached a maximum in the small secretory granules and labeling of medium (diameter between 1.1 and 2.0 m) and large (diameter over 2.0 m) granules was very low. At this postinjection interval the secretion product inside the lumen of the duct was already labeled. Between 1 and 72 h after injection the concentration of radioactivity in the small secretory granules decreased intensely while increasing in the medium and in the large ones. The concentration of fucose label reached a maximum in the medium secretory granules at 24 h and in the large ones at 72 h after injection. Additional experiments using mice previously injected with 4 intraperitoneal doses of ³H-fucose given 3 h apart demonstrated that the large granules undergo a very slow renewal. Some were found to be labeled as long as 28 days after administration of ³H-fucose. Recorded in this latter series of experiments was the labeling pattern of dense bodies that were regularly visualized in the cells lining the granular ducts. Their significance in the secretory process is discussed. In conclusion, newly synthesized glycoproteins are transferred from the Golgi apparatus to small secretory granules which carry a readily releasible pool of these macromolecules to the lumen of the duct. The small secretory granules also transfer newly synthesized glycoproteins to medium and large secretion granules which store a pool that is released very slowly. This characterizes the large secretory granules as the intracellular sites of storage of secretion products. The results of this investigation were correlated with the knowledge about the chemical composition of the different macromolecules that are known to be synthesized by the secretory cells of the granular ducts of the submandibular gland of the mouse.     

不同生长年限白头翁根的解剖结构与皂苷组织化学定位研究

皂苷类物质是白头翁的重要药用成分,主要存在于根内。研究白头翁根的解剖结构与皂苷类物质的关系有重要意义。利用解剖学和组织化学定位方法对不同生长龄白头翁根的解剖结构和皂苷的储藏场所进行了研究.结果表明:栓内层和次生韧皮部中的分泌细胞团、分泌腔和分泌道是白头翁根的主要结构特征,是皂苷类物质的主要贮存场所;而分泌腔和分泌道的数量及分泌道的宽窄与生长年限相关,4~5年生达到最大值。推测白头翁的栽培年限应为4年。     

Accurate root length and diameter measurement using NIH Image: use of Pythagorean distance for diameter estimation

This report describes an image analysis algorithm to estimate the length versus diameter of washed root samples accurately. Image analysis was performed using a Macintosh computer and the public domain NIH Image program. The binary image of the roots was processed to get the thinned image to calculate the length of the roots. The pixels of the root in a binary image were then stripped off from around the periphery based on the pixel's Pythagorean distance from the nearest background pixel. The length of the remaining root in each stripping off process was calculated after the image was thinned. Images (300 dpi) of copper wire of 0.23, 0.5, 1.0 mm diameter were analyzed for verification of the usefulness of the procedure. The results showed that more than 93% of the wires in each diameter wire were calculated to be in diameter classes including the true diameter and its adjoining classes: 93.6% of the wires of 0.23 mm diameter appeared in the 0.098–0.38 mm diameter classes, 96.19% of the wires of 0.5 mm diameter appeared in the 0.38–0.61 mm diameter classes, and 96.17% of the wires of 1 mm diameter appeared in the 0.85–1.08 mm diameter classes. The proposed method was tested for primary and secondary roots of water-cultured rice (Oryza sativa L.) and it was proven that the method could provide accurate length and diameter measurements for each root order. In addition, it was found that the method could provide the lengths of the thick primary, thin primary, and secondary roots. The effectiveness of applying sharpening for the grayscale image before making the binary image is also discussed.     

Relationships between FSH surges and follicular waves during the estrous cycle in mares

Individual follicles >/=15 mm were monitored daily by ultrasonography in 12 mares during the estrous cycle. Follicular waves were designated as major waves (primary and secondary) and minor waves based on maximum diameter of the largest follicle of a wave (major waves, 34 to 47 mm; minor waves, 18 to 25 mm). Dominance of the largest follicle of major waves was indicated by a wide difference (mean, 18 mm) in maximum diameter relative to the second largest follicle. Dominant follicles of primary waves (n=12) emerged (attained 15 mm) at a mean of Day 12 and resulted in the ovulations associated with estrus (ovulation=Day 0). The dominant follicle of a secondary wave (n=1) emerged on Day 2 and subsequently ovulated in synchrony with the dominant follicle of the primary wave, which emerged on Day 9. The largest follicles of minor waves (n=4) emerged at a mean of Day 5, reached a mean maximum diameter 3 days later, and subsequently regressed. There was a significant increase in mean daily FSH concentrations either 6 days (primary wave) or 4 days (minor waves) before the emergence of a wave. Mean concentrations of FSH decreased significantly 2 days after emergence of the primary wave. Divergence between diameter of the dominant and largest subordinate follicle of the primary wave was indicated by a significantly greater mean diameter of the dominant follicle than of the largest subordinate follicle 3 days after wave emergence and by the cessation of growth of the largest subordinate follicle beginning 4 days after the emergence of a wave. Surges of FSH were identified in individual mares by a cycle-detection program; surges occurred every 3 to 7 days. Elevated mean FSH concentrations over the 6 days prior to emergence of the primary wave was attributable to a significantly greater frequency of individual FSH surges before wave emergence than after emergence and to an increase in magnitude of peak concentrations of FSH associated with individual surges.     

The development of the sweat and sebaceous glands in the skin of the one-humped camel (Camelus dromedarius)

The sweat gland primordium appeared as a solid bud from the ental side of the epithelial cords of the growing hair follicles in the CRL 36 cm-fetus from the primary hair follicles and in the CRL 64 cm-fetus from the secondary ones. In the CRL 68 cm-fetus the glands were tubular with a slight curving of their lower third, which became convoluted at the CRL 83 cm-stage. At the latter age, clear myoepithelial cells were observed in the secretory portion as also in the duct. The buds of the sebaceous glands could be observed in the CRL 45 cm-fetus from the primary hair follicles and in the CRL 68 cm-fetus from the secondary ones. At the CRL 83 cm-fetal stage, the branched glands were elongated in shape and opened into the follicles by two separated ducts, united at the hair canal. The glands were in function nearly from its early development.     

Freeze-fracture surface of salivary glands of rat observed by scanning electron microscopy

The structure of parenchymal components of rat submandibular glands observed with scanning electron microscopy is presented. The glands were fixed, submerged in liquid nitrogen, cryofractured, dehydrated and critical-point-dried. The fracture surface displayed the acini, granular ducts and striated ducts. Acini exhibit a typical sponge-like pattern of irregular and empty cavities. Granular ducts are lined by bulging cells laden with numerous secretory granules. Their diameter ranged from 0.2 to 3.2 microns, with a mean value of 1.240.4 microns. Measurement of the actual granules was rendered possible by direct observation. Striated ducts exhibited a cribiform pattern near the basal cell region which corresponds to infoldings of the basal cell membrane.     

Morphometric measurement of local curvature of the alveolar ducts in lung mechanics

We have performed partial serial reconstructions of an acinus of the rabbit lung and determined the apparent existence of numerous heterogeneities in length, diameter, and local curvature in individual generation branches of the lung. We believe that structural changes during respiratory movements may include changes in the length and diameter of the whole duct. Alveoli are seen to be side differentiations secondary to the ducts in the gas-exchanging parenchyma of the rabbit lung. We have developed a technique for measuring local curvature in simple reconstructed ducts from the average of the integral curvation of the section contour. The contour curvature is measured from the chain code representation of the sampled contour from digital image analysis. The stereological requirements of an unbiased and random selection of contours is approached here by the random orientation of the individual alveoli of a single duct. Over 700 sections through the last four airway generations (alveolar ducts) at 3-micron intervals were analyzed. The average integral curvature ranges from 7.7 to 9.5 (mean 8.9) mm-1 for sixth- and seventh-generation branches from the start with volumes for the segments from 0.022 to 1.198 (mean 0.497) mm3.     

Kinase pathways in chemoattractant-induced degranulation of neutrophils: the role of p38 mitogen-activated protein kinase activated by Src family kinases

The aim of the present study was to investigate the role of tyrosine phosphorylation pathways in fMLP-induced exocytosis of the different secretory compartments (primary and secondary granules, as well as secretory vesicles) of neutrophils. Genistein, a broad specificity tyrosine kinase inhibitor, blocked the exocytosis of primary and secondary granules, but had only a marginal effect on the release of secretory vesicles. Genistein also inhibited the phosphorylation of extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinases (MAPK), raising the possibility that inhibition of ERK and/or p38 MAPK might be responsible for the effect of the drug on the degranulation response. Indeed, SB203580, an inhibitor of p38 MAPK, decreased the release of primary and secondary granules, but not that of secretory vesicles. However, blocking the ERK pathway with PD98059 had no effect on any of the exocytic responses tested. PP1, an inhibitor of Src family kinases, also attenuated the release of primary and secondary granules, and neutrophils from mice deficient in the Src family kinases Hck, Fgr, and Lyn were also defective in secondary granule release. Furthermore, activation of p38 MAPK was blocked by both PP1 and the hck-/-fgr-/-lyn-/- mutation. Taken together, our data indicate that fMLP-induced degranulation of primary and secondary granules of neutrophils is mediated by p38 MAPK activated via Src family tyrosine kinases. Although piceatannol, a reportedly selective inhibitor of Syk, also prevented degranulation and activation of p38 MAPK, no fMLP-induced phosphorylation of Syk could be observed, raising doubts about the specificity of the inhibitor.