A Double-Embedding Technique for Thin Tissue Membranes

A new double-embedding technique for thin tissue membranes is presented. This technique is useful for thin membranes such as mesenteric membranes from rodents, which usually measure only 10 μm in thickness. Several membranes are fixed and mounted on four needles located at the bottom of a plastic box. The box is filled with agarose at 50 C and then allowed to solidify. The agarose block is then removed, dehydrated in alcohol, cleared with Histo Petrol (isoparaffin hydrocarbons), permeated with paraffin and sectioned. The morphology is comparable to that obtained with methacrylate plastic embedding but is less time-consuming, less hazardous since no plastic hardener and activator are used and makes immunohistochemical studies easier.     

Effects of Inorganic Oxides,Sulfides, Chlorides,and Acid Chlorides on Metachromasia and Other Staining Properties

The ability of 17 inorganic compounds (POCl₃, PSC1₃, PC1₃, P₂O₅, P₂S₅, P₄S₃, P₄S₇, PC1₅, Sb₂O₅, As₂O₅, BiOC1₂, SeOC1₂, SO₂C1₂, Sb₂S₅, VOC1₂, SiC1₄ and CrO₂Cl₂) dissolved in pyridine or 2,2,4-trimethyl pentane, to enhance subsequent staining of tissue components with toluidine blue, phosphotungstic acid-hematoxylin (PTAH), leukofuchsin, and dihydroxydinaphthyl-disulfide (DDD) was studied. Eight of these compounds were also tested for ability to enhance staining with Alcian blue 8GN and Luxol fast blue MBS. Nine of the 17 compounds produced increased staining of certain tissue components with leukofuchsin, 13 with toluidine blue, 16 with PTAH, and 16 with DDD. The results suggest additional approaches to identification of tissue entities by induced metachromatic basophilia and leukofuchsin positivity as well as by the other stains studied, and also suggest a number of hitherto unstudied modes of reaction between the dyes used and reactive groups of tissue components. Many reactions of the compounds tested, with reactive groups known to be present in tissue components, are basecatalyzed, so that choice of solvent can influence the results obtained.     

生物组织背向SHG和TPEF的影响因素

由于生物组织的复杂性和多样性,以及样品制备等因素的影响,实验观察到的生物组织的背向二次谐波(second harmonic generation,SHG)和双光子激发荧光(two-photon excitation fluorescence,TPEF)效应的差异较大。以鼠尾组织作为实验对象,共40个切片,分为横向和纵向、HE染色和未染色四组,采用飞秒激光器作为激发光,用双光子激光扫描共焦显微镜(two-photon laser scanning confocal microscope,TPLSCM)观察和分析了样品在不同的制备方式、激发波长、激发功率、扫描深度等条件下的背向SHG和TPEF的变化曲线,讨论和比较了生物组织的背向SHG和TPEF的影响因素以及二者之间的异同,并尝试对实验现象做出了一定的解释。     

Effects of Various Gases on the Survival of Dried Bacteria During Storage

Salmonella newport and Pseudomonas fluorescens were dried together in papain digest broth and sucrose-glutamate, and stored in several gases at various water activities (a(w)) between 0.00 and 0.40 at 25 C for various periods up to 81 weeks. Both S. newport and P. fluorescens, dried in papain digest broth and stored in air, died rapidly if the conditions were very dry (0.00 a(w)) or moist (0.40 a(w)). Storage in carbon dioxide and argon gave greater survival than storage in air but lower survival than did storage in nitrogen or in vacuo. When the organisms were dried in a sucrose-glutamate mixture the differences between the gases were very small, and variations in residual water were less important. Of the inert gases, argon gave the best survival when the organisms were dried in papain digest broth, especially at 0.00 a(w); the survival in neon and krypton was lower and in xenon and helium it was much lower.     

蓝莓离体叶片胚状体高效发生及其组织学观察

以高灌蓝莓试管苗叶片为外植体、以改良WPM为基本培养基,研究了外源激素TDZ、ZT及其组合对离体叶片胚状体发生的影响,同时也探讨了蔗糖浓度、水解酪蛋白、椰汁等对胚状体发生、丛芽形成的影响.结果表明:不同浓度的外源激素TDZ、ZT及其组合对胚状体的发生频率、丛芽的形成和生长起重要作用;蔗糖浓度对胚状体的发生及丛芽的生长影响较大,而添加有机质则对胚状体的发生及丛芽的生长没有明显影响.适合高灌蓝莓叶片胚状体发生及成苗的培养基为WPM TDZ 0.04 mg/L ZT 0.25～2.0 mg/L 蔗糖20～40 g/L,而培养基WPM ZT 0.5～1.0 mg/L 蔗糖20 g/L适合于丛芽继代生长.组织学观察表明,蓝莓叶片胚状体发生主要起源于叶上表皮细胞和部分叶肉细胞,可能为多细胞起源,历经多细胞原胚、原球胚、梨形胚、心形胚、子叶胚等发育阶段,并能直接发育成苗.     

A tissue model for the study of cell proliferation in vitro

Summary A procedure for the cultivation of mesentery is described, in which the culture is fully representative of the tissue of origin. The intact mesenteric membrane—exposed to a minimum of trauma—was spread out over a hole in a filter paper strip in fluid medium and was cultivated free-hanging. Specimens from rats and guinea pigs were used. The organ culture model appears especially apt for cytochemical and proliferation studies. Proliferation variables based on Feulgen DNA analysis in individual, morphologically defined cells and on mitotic counting and radiochemical analysis were estimated. The tissue was fully viable in chemically defined growth medium and showed an almost unaltered light microscopical appearance after up to 52 hr in culture. Supported by the Swedish Cancer Society.     

Liquid Detergent in Aqueous Fixing Fluids, to Promote Rapid Wetting and Sinking of Plant Specimens

Plant tissue immersion for fixation is facilitated by the addition of a drop or two of liquid detergent to each 10 ml of fixing solution. Six fixatives: FAA, chrome-acetic, Craf I, III, V and Farmer's were tested with 1-7% additives of a liquid detergent (Joy was used). All were found to facilitate wetting and sinking of the specimens in the aqueous fixatives. Farmer's fixative (alcohol-acetic 3:1), however, did not require detergent. The detergent did not harm the tissue nor affect staining.     

An Evaluation of Various Fixing and Staining Procedures for Mitochondria in Plant Root Tissues

Mitochondria were stained intensely by a Regaud iron-hematoxylin procedure for roots fixed in formalin-sublimate or in Helly's fluid. Formalin-sublimate fixation required iodization during the staining sequence, but roots fixed in Helly's fluid were best iodized to remove mercurial precipitates before embedding in paraffin. Both methods required treatment with 1% KOH before immersion in the staining solution to remove RNA and produce pale cytoplasm. A third successful method was to postosmicate methacrylate-embedded roots after fixation in Hermann's fluid. Blackened mitochondria were produced by the postosmication and further staining was unnecessary. Fixation in Regaud's fluid did not give successful stains in any of the three methods tested. A prefixation treatment in quinone did not aid in obtaining sharply stained mitochondria of roots fixed in Bouin's fluid and stained with Heiden-hain's iron-hematoxylin.     

Effects of detergents on Ca2+-activated neural proteinase activity (calpain) in neural and non-neural tissue: A comparative study

Calcium activated neutral proteinase (mcalpain) activity was determined in brain and other tissue of rat. More than 60% of the brain mcalpain activity was present in the particulate fraction while only 30% was in cytosol. In contrast, particulate fractions of liver, kidney, muscle, and heart contained about 8–12% of tissue mcalpain activity while 88% was present in cytosol. Removal of the endogenous inhibitor calpastatin increased the tissue mcalpain activity severalfold. Triton X-100 and deoxycholate (DOC) stimulated the neural calpain activity by ten-fold while activity in non-neural tissue was unaffected. Incubation with other detergents, e.g. Triton N-57 and thioglucopyranoside, stimulated brain calpain activity five-fold while Brij-35 did not have any effect. Sodiumdodecylsulphate (SDS), on the other hand, inhibited the enzyme activity. Brain contained the lowest calpain activity compared to non-neural tissue. The calpain activity in muscle, kidney and heart was three-fold greater than liver. Immunoblot identification of the enzyme revealed that calpain was predominantly in the particulate fraction and less in cytosol of brain while it was present mainly in cytosol and less in the pellet fractions of non-neural tissue.     

Copper Concentration in Body Tissues and Fluids in Normal Subjects of Southern Poland

Data on the concentration of the elements in the human body are important, for example, to estimate the amounts required to maintain a good healthy state or find their connections with morbidity and mortality. In this paper, the concentration of copper (by flame atomic absorption spectrometry) in material obtained from autopsy cases of nonpoisoned people (n = 130), aged from 14 to 80 years, between 1990–2006, is presented. The following values were found (mean ± SD in micrograms of copper per gram or per milliliter): brain 3.32 ± 1.50 (n = 43), liver 3.47 ± 1.51 (n = 79), kidney 2.15 ± 0.90 (n = 76), stomach 1.10 ± 0.76 (n = 65), intestines 1.54 ± 1.19 (n = 25), lung 1.91 ± 1.30 (n = 27), spleen 1.23 ± 0.28 (n = 3), heart 3.26 ± 0.59 (n = 5), bile 3.60 ± 1.67 (n = 13), and blood 0.85 ± 0.19 (n = 73).     

采后芒果的后熟软化与贮藏淀粉变化之间的关系

研究了芒果后熟软化与贮藏淀粉变化之间的关系。结果表明,采后芒果淀粉的水解和消失,是使中果皮细胞壁失去支撑致使果实软化的重要原因之一。杀菌剂普克唑对芒果后熟软化和贮藏淀粉形态学的变化稍有延迟作用;2,4－D处理可以明显推迟后熟软化,减慢果实硬度的降低和淀粉的水解,而乙烯利处理则能催熟芒果,显著加速硬度丧失和淀粉的水解及其含量的减少。     

Total Relaxivities of Material Content in Various Cysts and Ameloblastoma: Implications for Discriminating Different Fluids

Nuclear magnetic resonance T ₁ and T ₂ relaxivities (r ₁ and r ₂) exhibit efficiency of a material to alter the relaxation rates (1/T ₁ and 1/T ₂), and they are being used for diagnostic purposes. The determination of total relaxivities (r _1t and r _2t) of cystic fluid content and ameloblastoma may therefore be useful for discriminative purposes. In order to determine what makes total relaxivities of hemorrhagic cysts, four sets of tubes containing pooled cyst were doped with increasing concentrations of iron, copper, albumin, and γ-globulins. These sets were replaced in a phantom together with six individual cysts and one ameloblastoma. The relaxation times were measured by magnetic resonance imaging operating at 1.5 T. The relaxivities of individual ions and proteins were determined from the slope of the relation between relaxation rates and concentration, while total relaxivities were determined by using the increases in relaxation rates and material content of cystic fluid (MC). Iron, copper, albumin, and γ-globulins were found to be the sources of r _1t and r _2t. Each of r _1t, r _2t, r _1tMC, r _2tMC, and r _2t/r _1t are distinctive parameters for each cystic category and ameloblastoma. Except for MC, the parameters measured for ameloblastoma are significantly smaller than those of cysts. The similarity of the present results to those used in clinical applications suggests that each of r _1t, r _2t, r _1tMC, r _2tMC, and r _2t/r _1t has an ability to discriminate various fluids and masses. The present work also suggests that r _1tMC, r _2tMC, and r _2t/r _1t can be determined in vivo.     

Effects of Different Ratios of Mixture of Starch and Casein,or Starch and Fat,in Diets on Lipogenesis in Rats

Studies were made on the incorporation of ¹⁴C of acetate-1-¹⁴C into the lipids of the liver and carcass, and changes in the concentrations of nucleotides, citric acid, and free fatty acids in the liver, using rats fed diets consisting of starch and casein, or starch and corn oil, at different ratios. Lipogenesis was stimulated with an increase in the content of starch both in the starch-casein and starch-corn oil diets. There was a rapid drop in lipogenesis in rats fed the diet with a little increment in the content of corn oil. Lipogenesis decreased gradually with an increment in the content of casein in the starch-casein diet. A positive correlation was found between lipogenesis and the level of ATP in the liver. The concentration of citric acid decreased with an increase in lipogenesis in the liver. Changes in dietary composition did not produce any significant alteration in the concentration of free fatty acids in the liver.     

兔VX2肝癌模型制作技术改良

目的探讨制作兔VX2肝癌模型的新技术、新方法。方法将新西兰兔30只随机等分为3组,分别为嵌插组、改良嵌插组和经皮穿剌组。分别按不同方法建立肝癌模型,于建模后第1、2、3周分别进行CT扫描,评估肿瘤大小。3周后处死动物进行解剖,观察肝脏成瘤及转移情况,比较各组转移率,并行常规病理组织学检查。结果 30只动物建模成功28只,成功率93%,经皮穿剌组有2只肝脏未见种植灶,嵌插组和改良嵌插组全部种植成功。改良嵌插组腹壁转移率显著低于嵌插组和经皮接种组。结论改良嵌插法3周内不会形成接种部位腹壁以及远处的转移,是一种理想的移植性肝癌模型建模方法。     

Effects of Various Amino Acids on Methionine-Induced Hyperhomocysteinemia in Rats

Rats were fed diets supplemented with 1% L-methionine with and without 2.5% various amino acids for 7 d to determine what amino acids other than glycine, serine, and cystine can suppress methionine-induced hyperhomocysteinemia. L-Glutamic acid, L-histidine, and L-arginine significantly suppressed methionine-induced enhancement of plasma homocysteine concentrations, but the mechanisms underlying the effect of these amino acids are thought not to be identical.     

冬凌草甲素抗炎作用参与多种疾病治疗的新进展

冬凌草甲素是冬凌草的主要活性成分,具有广泛的药理作用。研究表明,冬凌草甲素在炎症性疾病方面表现出较好的疗效。综述国内外关于冬凌草甲素抗炎作用参与多种疾病治疗的研究进展,并分析研究的方向,为研究与开发以冬凌草为原料的抗炎新药提供参考。     

Effects of Sunlight on Stained Sections Mounted in Various Media

Sections stained with haematoxylin and eosin showed maximum optical density (OD) at 536 nm, with a second peak at 600 nm. Sections stained with only eosin showed a peak at 536 nm, whilst those stained with haetnatoxylin showed a peak at 600 nm. Reduction in OD at these wavelengths was used to estimate fading of the staining. Direct sunlight reduced the OD of sections mounted in 22 different mounting media by 14 to 64% at 536 nm and 12 to 51% at 600 nm.     

壳聚糖在香蕉果实贮藏保鲜上的应用效果

以巴西香蕉果实为材料,研究了不同浓度（0、0．5％、1．0％、2．0％）的壳聚糖溶液对香蕉果实贮藏保鲜的影响。结果表明：壳聚糖处理可明显延缓香蕉果实的软化进程和病情指数的升高;同时维持了果实较低的细胞膜透性、多酚氧化酶（PPO）活性和较高的苯丙氨酸解氨酶（PAL）、β-1,3-葡聚糖酶（GUN）的活性。在四种处理中,以2％壳聚糖处理效果最好。表明壳聚糖对香蕉果实的贮藏保鲜效应可能与其调控病害相关酶的活性有关。