Contrasty Staining And Quantitative Determination Of Mucopolysaccharides On Filter Paper By Means Of Colloidal Iron

The contrast between the Prussian blue color of the mucopolysaccharide spots and paper background was increased by differentiation of the paper strips (dyed in acid colloidal iron solution made up in 60% ethanol) with thioglycolic acid. Thus the trivalent iron bound to the paper background was reduced but that adsorbed by the mucopolysaccharides was precipitated as ferric ammonium thioglycolate. This procedure was found to stain equally well acid (including sulfated) and neutral mucopolysaccharides, even though these varieties exhibited different staining properties by the periodic acid-Schiff and toluidine blue dyeing procedures. Staining of different depolymerization products of hyaluronate was little influenced by their chain lengths. Quantitative determination of the mucopolysaccharide content of the spots was performed by elution of ferrocyanide with hot sodium hydroxide and measuring of the Prussian blue color of the extracts, developed on acidification and addition of FeCl₃.     

Staining of Fungi With Saccharated Iron Oxide

A 0.2% aqueous solution of saccharated iron oxide with a pH of 10.8 is shown to be suitable for supravital staining of fungi. Colonies or fresh tissue materials presumed to contain fungi are immersed in the solution for 1-24 hr, fixed in a 10% neutral solution of buffered formalin about 1 hr, washed, and treated with 10% potassium ferrocyanide in 0.5 N, HC1. Cell membranes of hyphae, perithekia, and walls of sporangia appear dark blue; cytoplasm of the hyphae, sporangiospores and spores stain greenish blue or yellowish green.     

Culture And Staining of Pollen Grains of Ricinus communis

Germinating and growing pollen grains (male gametophytes) of Ricinus communis L. in liquid culture is achieved as follows: Pollen is collected over a 10-15 min period from mature anther clusters which have been removed from the male flowers and which have been kept at 25° C and 40-60% relative humidity. Samples weighing between 2.5 and 5.0 mg are brought as quickly as possible into a Desicote treated vial containing 17% sucrose and 30 ppm H₃BO₃ in boiled distilled water. The proportion (w/v) of pollen to culture solution should be 1:100. Shed pollen is kept in a humidity chamber whenever it is not being handled. The air in the culture vial is replaced by O₂ at the pressure of 1 atmosphere plus 5 lb and the sealed vials are shaken gently for 8-10 hr while partially immersed in a waterbath kept at 30° C. The pollen is fixed by the addition to the incubation suspension of an absolute alcohol-lactic acid (4:1) fixing fluid. The proportion used is 36 parts of fixing fluid to 1 part of culture solution. The fixed pollen can be stored in the fixative. Smears are prepared by applying single drops of the constantly agitated suspension of fixed pollen to a microscope slide. After each drop has spread out and dried, an additional drop is added until 10-20 have been applied. The preparations are stained by adding a drop of 1% acetic-orcein and are sealed with fingernail lacquer. The method is well adapted to the following types of studies: pollen germination, physiology of pollen tube growth, morphology of the male gametocyte, and physiology and cytology of the generative cell and nucleus.     

Combined In Situ Zymography, Immunofluorescence, And Staining Of Iron Oxide Particles In Paraffin-embedded, Zinc-fixed Tissue Sections

AbstractSuperparamagnetic iron oxide particles are used as potent contrast agents in magnetic resonance imaging. In histology, these particles are frequently visualized by Prussian blue iron staining of aldehyde-fixed, paraffin-embedded tissues. Recently, zinc salt-based fixative was shown to preserve enzyme activity in paraffin-embedded tissues. In this study, we demonstrate that zinc fixation allows combining in situ zymography with fluorescence immunohistochemistry (IHC) and iron staining for advanced biologic investigation of iron oxide particle accumulation. Very small iron oxide particles, developed for magnetic resonance angiography, were applied intravenously to BALB/c nude mice. After 3 hours, spleens were explanted and subjected to zinc fixation and paraffin embedding. Cut tissue sections were further processed to in situ zymography, IHC, and Prussian blue staining procedures. The combination of in situ zymography as well as IHC with subsequent Prussian blue iron staining on zinc-fixed paraffin-embedded tissues resulted in excellent histologic images of enzyme activity, protease distribution, and iron oxide particle accumulation. The combination of all three stains on a single section allowed direct comparison with only moderate degradation of fluorescein isothiocyanate-labeled substrate. This protocol is useful for investigating the biologic environment of accumulating iron oxide particles, with excellent preservation of morphology.     

妊娠期糖尿病患者体内铁负荷状态及氧化应激水平的研究

为了探讨铁代谢在妊娠期糖尿病(GDM)发病中的作用,对GDM患者体内铁负荷状态、氧化应激水平及抗氧化状态进行分析研究.在912例孕24～28周产前检查的孕妇中,按血糖筛查和糖耐量试验筛选出GDM孕妇32例为实验组,随机选择糖耐量正常孕妇26例作为对照组,分别测定两组孕妇的血红蛋白(Hb)等指标,以评价机体铁代谢状况;测...     

EZH2在胃癌组织中的表达及与Hp-L型感染关系的探讨

目的探讨EZH2在胃癌组织中表达的意义及与幽门螺杆菌L型(Helicobacter pylori-L,Hp-L)感染的关系。方法 (1)应用免疫组织化学Elivision法和革兰染色法检测80例胃癌组织及30例癌旁组织(对照组)中EZH2蛋白的表达和Hp-L型的感染情况;(2)采用逆转录多聚酶链反应(RT-PCR)技术检测30例新鲜胃癌组织及对应切缘正常胃黏膜组织(对照组)中EZH2的mRNA表达。结果胃癌组EZH2蛋白表达的阳性率高于对照组(P<0.05),且EZH2表达水平升高与肿瘤大小、浸润深度、淋巴结转移和TNM分期有关(P<0.05),与性别、年龄无关(P>0.05);RT-PCR显示,肿瘤组织、远端正常对照组织的EZH2表达量差异明显(P<0.01)。胃癌组Hp-L型检出率78.8%(63/80)与对照组23.3%(7/30)有显著性差异(P<0.05),与免疫组化Hp-L型抗原表达率73.8%(59/80)无显著性差异(P>0.05),Hp-L检出阳性率为71.3%(57/80);癌组中Hp-L型感染阳性组的EZH2表达阳性率高于Hp-L型阴性组(P<0.05),且Hp-L型阳性率和EZH2蛋白的表达呈正相关(r=0.250,P<0.05)。结论 EZH2蛋白和mRNA在胃癌中的表达增加,且与胃癌的浸润、转移相关,其机制可能与幽门螺杆菌L型(Hp-L型)感染有关。     

高山被孢霉的红四氮唑染色程度与菌体油脂中花生四烯酸含量的关系

研究了高山被孢霉菌体被红四氮唑(TTC)染色的条件,并探讨了染色程度与菌体油脂中花生四烯酸含量的关系．高山被孢霉的种子菌体被TTC染色的程度随种龄增加而增加,而种子中的油脂含量和油脂中的花生四烯酸含量也都随种龄增加而增加．在发酵过程中,菌体被TTC染色的程度和菌体中的油脂含量以及油脂中的花生四烯酸含量随培养时间增加而增加．三株具有相似油脂含量、不同花生四烯酸含量的高山被孢霉以及一株不产花生四烯酸的鲁氏毛霉的染色结果显示菌体被红四氮唑染色的程度与菌体油脂中的花生四烯酸含量具有正相关性．该发现有助于花生四烯酸高产菌的快速筛选．     

On the Iron Requirement of Lactobacilli Grown in Chemically Defined Medium

The iron requirement of four strains of lactobacilli (L. acidophilus, L. delbrueckii subsp. bulgaricus, L. plantarum, and L. pentosus) was studied in a synthetic medium under aerobic or anaerobic conditions. Effects of iron salt and iron-chelated compounds were tested on bacterial growth in manganese-free or -supplemented media. No significant growth stimulation was observed in any condition. These results support the absolute manganese requirement for optimum growth of lactobacilli and the needless incorporation of iron in growth media. Received: 5 November 1997 / Accepted: 20 January 1998     

健康学龄前儿童血浆氨基酸与血红蛋白含量

本文采用丹酰氯聚酰胺薄层分析法研究了６５名３—７岁健康儿童的血浆游离氨基酸和血红蛋白浓度。结果表明：（１）赖氨酸促进蛋白质合成不仅供机体生长需要,同样也供合成血红蛋白;（２）贫血前期已有血浆游离氨基酸的变化,继而出现贫血,提示儿童时期生长旺盛,应注重补充某些氨基酸消耗和补充维生素、叶酸、铁元素等造血原料。     

用单元内混合家系相关法计算遗传力时的抽样误差估计

对应用单元内混合家系相关法计算遗传力时的抽样误差估计问题进行了探讨,推导出的抽样误差估计公式可用于遗传力的显著性检验．     

The Stainability of Nerve Fibers by Protargol With Various Fixatives And Staining Technics

The influence of the commonly used tissue fixing reagents, individually and in various combinations, on subsequent staining by protargol was studied. The reagents used were formalin, formamide, picric acid, acetic acid, paranitrophenol, pyridine and chloral hydrate. Parraffin sections from intestine and peripheral nerve of cat, dog, monkey and rat were stained with protargol after fixation in various experimental mixtures of the fixing reagents. Satisfactory nerve stains of intestine were not obtained with regularity after any one fixing and staining procedure. (Good fixation and staining appeared to be influenced by properties inherent in the tissue itself and showed marked variations from animal to animal even in the same species.)Stains of nerve fibers in peripheral nerve trunks were much more easily obtained than in the intestine where good stains were sporadic and unpredictable. The use of a mixture of 0.5% protargol and 0.1% fast green FCF, is proposed as a silver-dye staining medium.     

Correlation Between Pancreatic Islet Uncoupling Protein-2 (UCP2) mRNA Concentration And Insulin Status in Rats

Hypothesizing that UCP2 may influence insulin secretion by modifying the ATP/ADP ratio within pancreatic islets, we have investigated the expression of intraislet UCP2 gene in rats showing insulin oversecretion (non-diabetic Zucker fa/fa obese rats, glucose-infused Wistar rats) or insulin undersecretion (fasting and mildly diabetic rats). We found that in Zucker fa/fa obese rats, hyperinsulinemia (1222 ± 98 pmol/1 vs. 128 ± 22 pmol/1 in lean Zucker rats) was accompanied by a significant increase in UCP2 mRNA levels. In rat submitted to a 5 day infusion with glucose, hyperinsulinemia (1126 ± 101 pmol/l vs. 215 ± 25 pmol/1 in Wistar control rats), coincided with an enhanced intraislet UCP2 gene expression, whereas a 8h or a 2 day-infusion did not induce significant changes in UCP2 mRNA expression. In rats made hypoinsulinemic and mildly diabetic by the injection of a low dose of streptozotocin, and in 4-day-fasting rats (plasma insulin 28 ± 5 pmol/1) UCP2 gene expression was sharply decreased. A 3-day-fast was ineffective. The data show the existence of a time-dependent correlation between islet mRNA UCP2 and insulin that may be interpreted as an adaptative response to prolonged insulin excess.     

Correlation of CrystaL Growth with the Staining of Axons by the Golgi Procedure

The surface of the specimens subjected to a modified Golgi technique (formalin fixed material; specimens in the following solution for 8-10 days at 27 C: 3% K₂Cr₂O₇, 100 ml, with the addition of 2.5-10 ml of 10% formalin and 6-25 gm of sucrose; then in 0.75% AgNO₃ for at least 2 days at 27 C) is sometimes covered with a fur of filamentous crystals and sometimes with a powdery precipitate of laminar crystals. In a series of experiments in which about 500 blocks of tissue were treated with variations of the staining procedure, good axonal stain was positively correlated with the appearance of filamentous crystals. These filaments have a thickness of 1-4 μ and grow at a rate of 160-330 μ/hr, reaching a length of 2-7 mm.     

Estimation of Water Diffusion Coefficients in Freeze-Concentrated Matrices of Sugar Solutions Using Molecular Dynamics: Correlation Between Estimated Diffusion Coefficients and Measured Ice-Crystal Recrystallization Rates

The diffusion coefficient of the water component in a freeze-concentrated matrix is a useful parameter for predicting and controlling the recrystallization rate of ice crystals in sugar solutions relevant to frozen desserts. Herein, application of molecular dynamics (MD) for estimating the water diffusion coefficient in a freeze-concentrated matrix of sugar solutions is described. Diffusion coefficients evaluated using MD with the optimized potentials for liquid simulations all atom force field and water models of three types (simple point charge, simple point charge extended, and transferable intermolecular potential-4 point) show a good positive linear relation with measured values, indicating that the MD methods used in this study are useful for predicting differences in water diffusion coefficients in a sugar freeze-concentrated matrix. Furthermore, similarly to measured values, the estimated diffusion coefficients show a good positive correlation with recrystallization rates of ice crystals, which suggests that MD is useful to predict differences in recrystallization rates of ice crystals in frozen sugar solutions.     

Differential Staining of Yeast for Purified Cell Walls, Broken Cells, And Whole Cells

Intact yeast cells are Gram positive but broken or disrupted cells are Gram negative. A counterstain with methyl green provides differential staining between cell wall and cytoplasm. The cells and cell fragments are dried on a slide and stained by a standard Gram stain. The preparation is then treated for 5 min with 1% phosphomolybdic acid, washed, and stained 0.5 min with 1% aqueous methyl green (unpurified by CHCl₃ extraction). Under these conditions whole, intact cells are dark purple or black, walls of broken cells and purified walls are light green, and the exposed cytoplasm stains light purple. All fractions can be easily differentiated.     

A Paradichlorobenzene, Saponin, Iron Mordant Sequence in the Staining of Meiotic Chromosomes of Ipomea

For the meiotic study of Ipomea spp., flower buds were stripped of the calyx and corolla and soaked in saturated aqueous paradichlorobenzene at about 28° C for 3 hr, transferred to acetic-alcohol (1:3) for 6 hr, then into 1% saponin solution and left overnight. They were mordanted in 1:3 acetic-alcohol saturated with ferric oxide for 24 hr and stained in a mixture of 1% aceto-carmine and 2% aceto-orcein with 1 N HCl in the proportion of 9:9:1. The preparations were mounted in 1% aceto-carmine for temporary use and made permanent by dehydration through the n-butanol schedule. The pollen mother cells had clear cytoplasm with deeply stained chromosomes.