Morphological heterogeneity of secretory granules of rat Clara cells: an immunocytochemical study.

The secretory granules of rat bronchiolar Clara cells were classified into different types by their ultrastructural appearances followed by immunocytochemistry using anti-rat 10 kDa Clara cell-specific protein (10 kDa CCSP) antibody. One predominant type was the oval to round granule (type A granule), of which the matrix was composed of a map-like mixture of electron-dense and less electron-dense material. Another predominant type was the rod-shaped granule (type B granule). The content of type B granules varied from a finely fibrillar (type B1 granule) to an electron-dense, rod-like (type B3 granule) structure. Various intermediate types (type B2 granule) between type B1 and B3 granules were also found. Small cytoplasmic vesicles were found occasionally in close proximity to type B2 or B3 granule. Another type of granule (type C granule) was large, up to 8 microns in diameter, and contained a moderately electron-dense amorphous matrix. Both type A and C granules stained at a similar density with the antibody. The nascent form of type A granules, which was found in the vicinity to the trans face of the Golgi apparatus, was also labeled. On the other hand, the labeling density of type B granules varied: type B1 granules were almost devoid of immunolabeling, whereas type B3 granules were intensely labeled. Type B2 granules stained with the antibody; however, the labeling density was less than that of type B3 granules. The small cytoplasmic vesicles of type B2 granules were labeled.(ABSTRACT TRUNCATED AT 250 WORDS)     

镉胁迫对秋华柳根系活力及其Ca、Mg、Mn、Zn、Fe积累的影响

以秋华柳为试验材料,采用水培试验方式,设置CK(0 mg·L^-1 Cd²⁺)、T₁(2 mg·L^-1 Cd²⁺)、T₂(10 mg·L^-1Cd²⁺)、T₃(20 mg·L^-1Cd²⁺)、T₄(50 mg·L^-1Cd²⁺)5种镉处理浓度,通过对秋华柳根系活力,叶、韧皮部、木质部和根部的镉含量,以及Ca、Mg、Mn、Zn、Fe 5种常规金属元素含量的测定,研究了不同浓度镉胁迫下秋华柳根系活力及Ca、Mg、Mn、Zn、Fe 等金属元素含量的变化.结果表明:1)秋华柳根系活力随着镉处理浓度的增加而逐渐下降,当镉浓度≥10 mg·L^-1时,根系活力与对照相比显著下降.2)随着镉处理浓度的增加,秋华柳叶中Fe的积累受到显著抑制;韧皮部Mg、Mn、Fe的积累受到显著抑制;木质部Ca、Mg、Mn、Zn、Fe 等5种常规金属元素的积累无显著性差异;根部5种常规金属元素的吸收和积累受到显著抑制,表现出镉对其他金属元素积累的拮抗作用.其中,50 mg·L^-1的镉胁迫下,根部Zn的积累量降幅最大,受Cd的抑制最明显.3)各处理组Fe的转移系数与对照相比均无显著性差异;Ca、Mg、Mn、Zn的转移系数均高于对照,且在一定的镉处理水平上差异显著.4)镉胁迫下,秋华柳根部镉的积累量与Ca的积累量呈显著负相关,与其他4种常规金属元素的积累量呈极显著负相关,说明根部常规金属元素的变化可作为秋华柳受镉毒害程度的指示之一.     

Morphological heterogeneity of secretory granules of rat Clara cells: an immunocytochemical study

Summary The secretory granules of rat bronchiolar Clara cells were classified into different types by their ultrastructural appearances followed by immunocytochemistry using anti-rat 10 kDa Clara cell-specific protein (10 kDa CCSP) antibody. One predominant type was the oval to round granule (type A granule), of which the matrix was composed of a map-like mixture of electron-dense and less electron-dense material. Another predominant type was the rod-shaped granule (type B granule). The content of type B granules varied from a finely fibrillar (type B1 granule) to an electron-dense, rod-like (type B3 granule) structure. Various intermediate types (type B2 granule) between type B1 and B3 granules were also found. Small cytoplasmic vesicles were found occasionally in close proximity to type B2 or B3 granule. Another type of granule (type C granule) was large, up to 8 m in diameter, and contained a moderately electron-dense amorphous matrix. Both type A and C granules stained at a similar density with the antibody. The nascent form of type A granules, which was found in the vicinity to the trans face of the Golgi apparatus, was also labeled. On the other hand, the labeling density of type B granules varied: type B1 granules were almost devoid of immunolabeling, whereas type B3 granules were intensely labeled. Type B2 granules stained with the antibody; however, the labeling density was less than that of type B3 granules. The small cytoplasmic vesicles of type B2 granules were labeled. From these findings, it is suggested that the granules of rat Clara cells consist of two types of granules of distinct origin; one appears to derive from condensing vacuoles of Golgi origin, whereas the other may be formed by membranefusions with small cytoplasmic vesicles of unknown source.     

Elemental composition of secretory granules in pancreatic islets of Langerhans.

We have characterized, by electron probe microanalysis, rapidly frozen cultured rat islets at the level of individual secretory granules. Elemental analysis of thin, dried cryosections showed that beta granules could be distinguished by high Zn, Ca, and S, whereas non-beta (mainly alpha) granules contained elevated P and Mg. Although a single granule type predominated in a particular cell, some rebel granules were found in A cells that had the compositional fingerprint of B cell granules. Zn, which was found in millimolar concentrations in B cell granules, was considered a marker for the insulin storage complex. The data indicate that non-B islet cells in the adult pancreas may produce insulin-containing organelles and that, when glucagon and insulin are coexpressed, these hormones are packaged in separate granules.     

水稻锌铁转运蛋白ZIP基因家族研究进展

锌(zinc, Zn)和铁(iron, Fe)是水稻(Oryza sativa L.)生长必需的矿质元素,也是人体必需的微量元素。水稻体内Zn、Fe含量维持在适宜水平有利于提高其产量和品质,提高稻米中Zn、Fe含量能够在一定程度上解决人体Zn、Fe营养缺乏的问题。因此,研究水稻中Zn和Fe等微量元素转运蛋白的具体功能对于提高水稻产量和稻米品质具有重要意义。锌铁转运蛋白(zinc-regulated transporters and iron-regulated transporter-like protein, ZIP)负责Zn和Fe等离子的吸收、转运和分配,是维持水稻中Zn和Fe平衡的重要转运蛋白,其表达水平受Zn和Fe水平影响。ZIP基因家族在自然群体中具有丰富的等位变异,而且某些单倍型存在明显的籼粳分化,这可能造成了不同品种间籼、粳稻中Zn和Fe积累的差异。目前,已有大量关于ZIP基因家族的研究,但只有OsZIP3的作用机制研究的较为清楚。另外,对Zn、Fe在籽粒中的积累机制研究和自然群体中ZIP基因的等位变异研究还不够深入。因此,ZIP转运蛋白家族仍存在较大的研究空间。本文详细介绍了ZIP转运蛋白在水稻体内的亚细胞定位、表达模式、转运机制以及在自然群体中的等位变异等,以期为研究水稻稻米微量元素的积累提供理论基础,为提高稻米品质提供借鉴。     

Taxonomic units, civilization, volcanism: Their influence on the chemical composition of ostracod carapaces (Kagoshima Bay, Kyushu Island, Japan)

To test the influence of both civilization and volcanism on the chemical composition of ostracod carapaces, 32 well-preserved valves from Kagoshima Bay (Kyushu Island, South Japan) were analysed by means of spark source mass spectrometry. In Kagoshima Bay, the extent of pollution corresponds to human activities. Kagoshima Bay is also an area of important volcanic activity: emissions from Sakurajima volcano are very frequent. The species analysed were Argilloecia hanaii, Callistocythere undulatifacialis, Pontocythere subjaponica and Loxoconcha tosaensis. Twenty-three chemical elements were detected. Among them, 11 were always above the limit of detection: Al, B, Cl, F, Fe, K, Na, P, S, Si and Zn. The correlation between Zn and Fe was characteristic of the volcanic environment. Two discriminant analyses were performed using either the location area or the species as grouping variables. An association between Zn and S is possible; it could be related to volcanic activity. The chemical composition of carapaces is ascribed to taxonomic units rather than the environment.     

Aerobic granules: a novel zinc biosorbent

AIMS: Aerobic granules are aggregates with a compact and porous microbial structure. In view of the potential use of aerobic granules as biosorbents for Zn(II) removal from industrial wastewater, this study investigated the effects of initial Zn(II) and aerobic granule concentrations on the kinetics of Zn(II) biosorption on the aerobic granule surface. METHODS AND RESULTS: Acetate-fed aerobic granules with a mean diameter of 1.0 mm were used as biosorbents. Results showed that the kinetics of Zn(II) biosorption on the aerobic granule surface were related to both initial Zn(II) and granule concentrations. It was found that the real driving force for Zn(II) biosorption on the aerobic granule surface could be described by the ratio of initial Zn(II) concentration (Co) to initial granule concentration (Xo), rather than individual Co or Xo. The Co/Xo ratio provides a unified basis for interpretation of the biosorption data obtained under different initial conditions. The maximum biosorption capacity of Zn(II) by aerobic granules was 270 mg g(-1). CONCLUSIONS: It appears that the aerobic granule can be used as an effective biosorbent for efficient removal of Zn(II) or other types of heavy metals from industrial wastewater. SIGNIFICANCE AND IMPACT OF THE STUDY: This study could lead to the development of a novel granular biosorbent for the removal of heavy metals from wastewater. A simple and compact aerobic granule-based biosorber could be expected.     

Use of granules and free salts for Fe and Zn fortification of leafy vegetables: Improvements in trace element bioaccessibility and fulfillment of Dietary Reference Intakes

BackgroundLeafy vegetables represent an excellent dietary source of trace elements such as Fe and Zn. Nevertheless, Fe and Zn bioaccessibility can lessen due to a high concentration of anti-nutritional compounds. The encapsulation of Fe and Zn salts as granules could be used to fortify these leafy vegetables.MethodThree leafy vegetables, spinach, Swiss chard and Ethiopian mustard were fortified with iron sulfate and zinc sulfate as granules and free salts in order to test the improvements in the bioaccessibility and fulfillments of DRIs. Fe and Zn granules were prepared in a fluidized bed granulator. A probabilistic analysis was performed, using experimental data, to assess bioaccessible intake and fulfillments of DRIs in European populations.ResultsFe contents ranged between 4.8 mg/100 g of Ethiopian mustard to 157.4 mg/100 g of spinach. Fe and Zn bioaccessibility percentages were low for Swiss chard and spinach without fortification. Fortification with granules improved Fe bioaccessibility of these latter vegetables (196 and 223 mg/100 g). Zn contents in samples without fortification ranged between 2.3 mg/100 g for Ethiopian mustard and 7.4 mg/100 g for spinach. Zn fortification as granules improved Zn bioaccessibility for the three vegetables studied. Thus, Zn bioccessible concentrations ranged between 17.4 and 108 mg/100 g for the solubility assay and between 5.9 and 31.1 mg/100 g for the dialyzability assay. Besides, the probability analysis showed that fortification had a better performance in meeting DRIs for those populations with higher consumption levels of leafy vegetables.ConclusionsThe probability analysis demonstrated that fortification can be a suitable strategy to meet DRIs for both trace elements, which was especially remarkable for Fe. Fortification with granule was more effective in most the cases, although for Ethiopian mustard, free salt of Fe showed a better performance.     

Simultaneous iron, zinc, sulfur and phosphorus speciation analysis of barley grain tissues using SEC-ICP-MS and IP-ICP-MS

The increasing prevalence of iron (Fe) and zinc (Zn) deficiencies in human populations worldwide has stressed the need for more information about the distribution and chemical speciation of these elements in cereal products. In order to investigate these aspects, barley grains were fractionated into awns, embryo, bran and endosperm and analysed for Fe and Zn. Simultaneously, phosphorus (P) and sulfur (S) were determined since these elements are major constituents of phytic acid and proteins, respectively, compounds which are potentially involved in Fe and Zn binding. A novel analytical method was developed in which oxygen was added to the octopole reaction cell of the ICP-MS. This approach greatly improved the sensitivity of sulfur, measured as (48)SO(+). Simultaneously, Fe was measured as (72)FeO(+), P as (47)PO(+), and Zn as (66)Zn(+), enabling sensitive and simultaneous analysis of these four elements. The highest concentrations of Zn, Fe, S and P were found in the bran and embryo fractions. Further analysis of the embryo using SEC-ICP-MS revealed that the speciation of Fe and Zn differed. The majority of Fe co-eluted with P as a species with the apparent mass of 12.3 kDa, whereas the majority of Zn co-eluted with S as a 3 kDa species, devoid of any co-eluting P. Subsequent ion pairing chromatography of the Fe/P peak showed that phytic acid (myo-inositol-1,2,3,4,5,6-hexakisphosphate: IP(6)) was the main Fe binding ligand, with the stoichiometry Fe(4)(IP(6))(18). When incubating the embryo tissue with phytase, the enzyme responsible for degradation of phytic acid, the extraction efficiency of both Fe and P was doubled, whereas that of Zn and S was unaffected. Protein degradation on the other hand, using protease XIV, boosted the extraction of Zn and S, but not that of Fe and P. It is concluded that Fe and Zn have a different speciation in cereal grain tissues; Zn appears to be mainly bound to peptides, while Fe is mainly associated with phytic acid.     

秸秆还田配施中微量元素对农田土壤有机碳固持的影响

为研究秸秆还田配施中量元素(S)和微量元素(Fe和Zn)对粮田土壤有机碳固持的影响,进行了为期52 d的室内玉米秸秆腐解培养试验. 结果表明:秸秆腐解过程中分别添加S、Fe和Zn元素,均提高了微生物生物量碳(MBC)及土壤CO₂-C矿化速率,52 d腐解培养结束后,CO₂-C的累积矿化量显著提高,但土壤有机碳含量并未显著降低;3种元素中,添加Fe或Zn的处理提高了土壤惰性碳库、惰性碳库比例及土壤有机碳表观平衡,有利于土壤有机碳固持,而添加S的处理却降低了惰性有机碳比例及土壤有机碳表观平衡,不利于有机碳固持. 因此,在施N、P肥基础上,秸秆还田添加S、Fe或Zn均能促进土壤有机碳的矿化进程,但添加Fe或Zn可使更多有机碳固持于土壤中,添加S不利于土壤有机碳的固持.     

Presence of Active Hatching Enzyme in the Secretory Granule of Prehatching Medaka Embryos

Secretory granules of hatching gland were isolated from a 0.3 M sucrose homogenate of whole medaka embryos at prehatching stage by differential centrifugation, followed by a Percoll density gradient centrifugation. The obtained preparation was almost free of melanosomes and composed exclusively of the secretory granules of hatching gland (hatching enzyme granules), as judged by morphological as well as enzymological criteria.
The aqueous extracts of the purified secretory granules showed a specific choriolytic activity as high as about 40 times that of a partially purified secretory granule preparation, P_1,000, and represented a single protein band with molecular weight of about 21,000 on SDS-polyacrylamide gel electrophoresis. It was also revealed that a major component of the hatching enzyme preparation (P II–0.3 enzyme, 13) purified from the hatching liquid was identical with the 21,000 molecular weight band.
These results suggest that the hatching enzyme is present in the secretory granules of prehatching embryos in an active molecular form.     

Intracellular inclusions of uncultured magnetotactic bacteria.

Magnetotactic bacteria produce magnetic crystals in organelles called magnetosomes. The bacterial cells may also have phosphorus-containing granules, sulfur globules, or polyhydroxyalkanoate inclusions. In the present study, the ultrastructure and elemental composition of intracellular inclusions from uncultured magnetotactic bacteria collected in a marine environment are described. Magnetosomes contained mainly defect-free, single magnetite crystals with prismatic morphologies. Two types of phosphorus-containing granules were found in magnetotactic cocci. The most common consisted of phosphorus-rich granules containing P, O, and Mg; and sometimes also C, Na, Al, K, Ca, Mn, Fe, Zn, and small amounts of S and Cl were also found. In phosphorus-sulfur-iron granules, P, O, S, Na, Mg, Ca, Fe, and frequently Cl, K, and Zn, were detected. Most cells had two phosphorus-rich granules, which were very similar in elemental composition. In rod-shaped bacteria, these granules were positioned at a specific location in the cell, suggesting a high level of intracellular organization. Polyhydroxyalkanoate granules and sulfur globules were less commonly seen in the cells and had no fixed number or specific location. The presence and composition of these intracellular structures provide clues regarding the physiology of the bacteria that harbor them and the characteristics of the microenvironments where they thrive.     

Nocardioides halotolerans sp. nov., isolated from soil on Bigeum Island, Korea

A strictly aerobic, Gram-positive, motile, coccoid-shaped, halotolerant actinobacterium (10% NaCl, w/v), designated MSL-23^T, was isolated from a soil sample on Bigeum Island, Korea. Results of 16S rRNA gene sequence analysis indicated that the isolate belonged to the genus Nocardioides, with the highest sequence similarity (95.63%) being to Nocardioides kribbensis KCTC 19038^T. The major menaquinone was MK-8(H₄), and the predominant cellular fatty acids were i-C_16:0, ai-C_17:0, C_18:1 ω9c and 10-methyl-C_16:0. The DNA G+C content was 69.7 mol%. The 16S rRNA gene sequence of strain MSL 23^T and its chemotaxonomic properties showed it to be unique in the genus Nocardioides. Phenotypic characteristics distinguished strain MSL-23^T from other Nocardioides species. On the basis of the phenotypic, chemotaxonomic and phylogenetic data strain MSL-23^T represents a novel species, for which the name Nocardioides halotolerans sp. nov. is proposed, with MSL-23^T (=KCTC 19274^T=DSM 19273^T) as the type strain.     

物种和基物偏好对河北太行山区六种地衣的元素含量的影响

为了解物种和基物偏好对地衣元素水平的影响,测定了采集于河北太行山区6个地点的6种地衣体内的16种元素（Al、Ba、Cd、Cr、Cu、Fe、Mn、Mo、Na、P、S、Sc、Sr、Ti、V和Zn）的含量。这6种地衣包括3种石生地衣（鸡冠胶衣Collema cristatum、中国石黄衣Xanthoria mandschurica和淡腹黄梅Xanthoparmelia mexicana）,2种土生地衣（莲座石蕊Cladonia pocillum和石果衣Endocarpon pusillum）,及1种树生地衣（毛边黑蜈蚣衣Phaeophyscia hispidula）。主成分分析（PCA）和费里德曼双向秩方差分析（Friedman检验）结果显示,物种和基物偏好对地衣体内元素含量的影响显著,而地点的影响不显著。树生地衣累积的大气源元素（P、S和Zn）高于土生和石生地衣。土生地衣体内的地壳源元素（Al、Ba、Fe、Mn、Mo、Na、Sc、Sr、Ti和V）和大气源元素Cr的含量最高。石生地衣的大气源元素（P、S和Zn）含量近于土生地衣,但地壳源元素含量较低;3种石生地衣在这些元素的含量方面差异不大。这些结果表明以地衣化学组成监测大气元素沉降的研究需慎重考虑物种及基物偏好的影响。     

Dynamic characteristics of metal element content and return of Casuarina equisetifolia litter at different distances to the coastline

精确估算生态系统内部环境梯度引起的养分循环差异, 对评估其生态功能十分重要。为探讨不同距海生境对木麻黄(Casuarina equisetifolia)凋落叶金属元素含量及养分归还动态的影响, 以福建省惠安赤湖国有防护林场木麻黄凋落叶为研究对象, 按照离海由近及远的顺序设置5个距离(T₁到T₅), 收集凋落叶测定金属元素含量。结果表明: 1)海岸梯度环境对木麻黄凋落叶各元素含量及归还量具有显著影响, 由基干林带向林内元素含量、归还量呈线性规律, 总体上近海高于远海, 按照离海由近及远元素含量逐渐降低, 铁(Fe)、铜(Cu)、锌(Zn)和锰(Mn)在T₄后上升, 归还量变化趋势与含量相似。2)凋落叶元素含量在不同月份存在显著差异, Na、Fe、Cu、Zn具有相似性, 6月、11月存在明显低谷; Mn在2至4月下降后波动上升。归还量总体表现为钠(Na)、Cu相似, 5、8、次年1月出现峰值;Mn、Fe、Zn相似, 5、8、11、次年1月出现峰值。3)土壤Na含量按照离海由近及远逐渐降低, 其他元素波动变化, 各距离之间差异不明显。4)凋落叶Na-Fe含量存在显著正相关关系, Na-Zn、Fe-Cu、Fe-Zn含量存在极显著正相关关系; 土壤与凋落叶相同元素含量间存在正相关关系, Na和Cu达到显著水平。究其原因, 叶片元素含量差异受叶凋落时间差异造成的重吸收差异影响大; 月间差异因植物生命活动对元素需求量存在差异; 归还量差异与凋落叶量及元素含量有关。     

不同海岸距离上木麻黄凋落叶金属元素含量及归还量动态特征

精确估算生态系统内部环境梯度引起的养分循环差异, 对评估其生态功能十分重要。为探讨不同距海生境对木麻黄(Casuarina equisetifolia)凋落叶金属元素含量及养分归还动态的影响, 以福建省惠安赤湖国有防护林场木麻黄凋落叶为研究对象, 按照离海由近及远的顺序设置5个距离(T₁到T₅), 收集凋落叶测定金属元素含量。结果表明: 1)海岸梯度环境对木麻黄凋落叶各元素含量及归还量具有显著影响, 由基干林带向林内元素含量、归还量呈线性规律, 总体上近海高于远海, 按照离海由近及远元素含量逐渐降低, 铁(Fe)、铜(Cu)、锌(Zn)和锰(Mn)在T₄后上升, 归还量变化趋势与含量相似。2)凋落叶元素含量在不同月份存在显著差异, Na、Fe、Cu、Zn具有相似性, 6月、11月存在明显低谷; Mn在2至4月下降后波动上升。归还量总体表现为钠(Na)、Cu相似, 5、8、次年1月出现峰值;Mn、Fe、Zn相似, 5、8、11、次年1月出现峰值。3)土壤Na含量按照离海由近及远逐渐降低, 其他元素波动变化, 各距离之间差异不明显。4)凋落叶Na-Fe含量存在显著正相关关系, Na-Zn、Fe-Cu、Fe-Zn含量存在极显著正相关关系; 土壤与凋落叶相同元素含量间存在正相关关系, Na和Cu达到显著水平。究其原因, 叶片元素含量差异受叶凋落时间差异造成的重吸收差异影响大; 月间差异因植物生命活动对元素需求量存在差异; 归还量差异与凋落叶量及元素含量有关。     

An electron microscopic study on the effects of reserpine on the subclavian glomera of the rabbit.

Young male and female New Zealand white rabbits were given a daily subcutaneous injection of reserpine (Serpasil, Ciba; 3 mg/kg) for two days and were sacrificed 24 hours after the last injection. The subclavian glomera (aortic bodies) were processed for electron microscopy to determine the effects of this biogenic amine depleting agent on the electron-opaque cytoplasmic granules of the parenchymal type I cells. Observations of glutaraldehyde-osmium tetroxide fixed glomera from reserpinized animals showed a slight decrease in granule density of the type I cells. Glomera fixed in glutaraldehyde and incubated in potassium dichromate (pH 4.1) demonstrated a reduction in granule opacity following reserpine treatment. Control glomera incubated in potassium dichromate displayed electron-opaque granules. These results indicate that reserpine does deplete the amines without granule disappearance or changes in granule population. The positive reaction of the control tissue granules to potassium dichromate incubation suggests that the predominant biogenic amines in the electron-opaque granules are unsubstituted monoamines. Persistence of the opaque granules following reserpinization and glutaraldehyde-osmium tetroxide double fixation, may be due to amine-binding protein within the granules. The mode of granule depletion could not be ascertained with certainty.     

X-ray microanalysis of biological specimens by high voltage electron microscopy

For the purpose of analyzing and imaging chemical components of cells and tissues at the electron microscopic level, 3 fundamental methods are available, chemical, physical and biological. Among the physical methods, two methods qualifying and quantifying the elements in the structural components are very often employed. The first method is radioautography which can demonstrate the localization of radiolabeled compounds which were incorporated into cells and tissues after the administration of radiolabeled compounds. The second method is X-ray microanalysis which can qualitatively analyze and quantify the total amounts of elements present in cells and tissues. We have developed the two methodologies in combination with intermediate high or high voltage transmission electron microscopy (200–400 kV) and applied them to various kinds of organic and inorganic compounds present in biological materials. As for the first method, radioautography, I had already contributed a chapter to PHC (37/2). To the contrary, this review deals with another method, X-ray microanalysis, using semi-thin sections and intermediate high voltage electron microscopy developed in our laboratory.

X-ray microanalysis is a useful method to qualify and quantify basic elements in biological specimens. We first quantified the end-products of histochemical reactions such as Ag in radioautographs, Ce in phosphatase reaction and Au in colloidal gold immunostaining using semithin sections and quantified the reaction products observing by intermediate high voltage transmission electron microscopy at accelerating voltages from 100 to 400 kV. The P/B ratios of all the end products Ag, Ce and Au increased with the increase of the accelerating voltages from 100 to 400 kV. Then we analyzed various trace elements such as Zn, Ca, S and Cl which originally existed in cytoplasmic matrix or cell organelles of various cells, or such elements as Al which was absorbed into cells and tissues after oral administration, using both conventional chemical fixation and cryo-fixation followed by cryo-sectioning and freeze-drying, or freeze-substitution and dry-sectioning, or freeze-drying and dry-sectioning producing semithin sections similarly to radioautography. As the results, some trace elements which originally existed in cytoplasmic matrix or cell organelles of various cells in different organs such as Zn, Ca, S and Cl, were effectively detected. Zn was demonstrated in Paneth cell granules of mouse intestines and its P/B ratios showed a peak at 300 kV. Ca was found in human ligaments and rat mast cells with a maximum of P/B ratios at 350 kV. S and Cl were detected in mouse colonic goblet cells with maxima of P/B ratios at 300 kV. On the other hand, some elements which were absorbed by experimental administration into various cells and tissues in various organs, such as Al in lysosomes of hepatocytes and uriniferous tubule cells in mice was detected with a maximum of P/B ratios at 300 kV.

From the results, it was shown that X-ray microanalysis using semi-thin sections observed by intermediate high voltage transmission electron microscopy at 300–400 kV was very useful resulting in high P/B ratios for quantifying some trace elements in biological specimens. These methodologies should be utilized in microanalysis of various compounds and elements in various cells and tissues in various organs.     

Low-molecular-weight constituents of isolated insulin-secretory granules. Bivalent cations, adenine nucleotides and inorganic phosphate.

The concentrations of Zn2+, Ca2+, Mg2+, Pi and adenine nucleotides were determined in insulin-secretory granules prepared from a transplantable rat insulinoma. Differential and density-gradient centrifugation analyses revealed that Zn2+ in this tissue was principally localized in the secretory granule, a second major fraction being found in association with cytosolic proteins. Pi was principally recovered in the latter fraction, whereas Ca2+ and Mg2+ were more widely distributed. Intragranular ion-distribution experiments suggested that Zn2+ was complexed mainly to insulin and its precursor forms and remained in the granule in an insoluble state. The Zn2+/insulin ratio (0.54) was greater than that expected for insulin molecules having two centrally co-ordinated Zn2+ atoms/hexamer, but less than the maximal Zn2+-binding capacity of the molecule. Most of the granular Ca2+, Mg2+ and Pi was released in a soluble form when granules were disrupted by sonication. Simulation in vitro of the ionic composition of the granule suggested that up to 90% of its Ca2+ was complexed to Pi and adenine nucleotides. Granular macromolecules also bound Ca2+, as shown by equilibrium-dialysis studies of granule lysates. However, such binding was displaced by Mg2+. Examination of the efflux of Ca2+ from granules incubated in iso-osmotic suspensions at 37 degrees C suggested that the passive permeability of the granule membrane to Ca2+ was very low. Nevertheless, more than 50% of the granular Ca2+ was rapidly released in an ionized form on hypo-osmotic or detergent-induced disruption of the granule membrane. This may represent a potentially mobilizable pool of Ca2+ in vivo.     

风车草和香根草在人工湿地中迁移养分能力的比较研究

为研究风车草(Cyperus alternifolius)和香根草(Vetiveria zizanioides)迁移养分的能力,建立17.0m²风车草潜流式人工湿地和13.3 m²香根草潜流式人工湿地处理猪场废水,在四个季节末测定植物生物量和组织氮、磷、铜、锌含量.结果表明,香根草地下部生物量大于风车草,地上部生物量则是风车草大于香根草.风车草年地上部收获量为3406.47 g·m^-2,比香根草的1483.88 g·m^-2高2.3倍;风车草的氮含量为22.69 mg·g^-1,比香根草的15.44 mg·g^-1高7.25 mg·g^-1;风车草的磷含量为6.09 mg·g^-1,比香根草的5.47 mg·g^-1高0.62 mg·g^-1.植株含铜、锌量风车草略比香根草高.风车草每年迁移N 68.72 g·m^-2和P18.49 g·m^-2,香根草迁移N 8.93 g·m^-2和P 3.69 g·m^-2.风车草人工湿地每年由植物迁移的氮、磷、铜、锌比香根草高4～7倍.