Calcium peroxide as a seed coating material for padi rice

Summary Measurement of oxygen uptake by germinating rice seed (Oryza sativa L) suggests that oxygen requirement is independent of temperature of incubation. However, the rate of oxygen consumption is dependent upon incubation temperature and, after an initial lag phase, is exponential with time. Although rice seed can germinate and grow at low oxygen concentrations, germination is poor and seedlings exhibit low vigour. An oxidized zone may be observed around the seed when sown in an anoxic environment but coated with a layer of calcium peroxide. The seed germinates readily and develops normally when a sufficient level of calcium peroxide is used.     

Laser-assisted preparation of single cells from stained histological slides for gene analysis

 Individual cells are prepared from histological tissue sections of routinely formalin-fixed and paraffin-embedded tissues using an ultraviolet laser micromanipulator. This technology, in combination with polymerase chain reaction (PCR)-based gene analysis, will enable researchers to routinely detect a variety of nucleic acid abnormalities underlying cancer, infection, and genetic disease with previously unknown sensitivity: at the single cell level. The utility of this technique is demonstrated by PCR amplification and sequencing of the E-cadherin gene, which codes for a homophilic cell-to-cell adhesion molecule, in early gastric carcinomas of the diffuse type of Lauren’s classification. The main characteristics of the laser-assisted microdissection technique are high precision without contamination and easy application. The assignment of individual gene sequences to single cells will now provide a direct link between molecular biology on the one hand and histology and pathology on the other. Accepted: 18 August 1997     

Dendrimeric coating of glass slides for sensitive DNA microarrays analysis

Successful use and reliability of microarray technology is highly dependent on several factors, including surface chemistry parameters and accessibility of cDNA targets to the DNA probes fixed onto the surface. Here, we show that functionalisation of glass slides with homemade dendrimers allow production of more sensitive and reliable DNA microarrays. The dendrimers are nanometric structures of size-controlled diameter with aldehyde function at their periphery. Covalent attachment of these spherical reactive chemical structures on amino-silanised glass slides generates a reactive ~100 Å layer onto which amino-modified DNA probes are covalently bound. This new grafting chemistry leads to the formation of uniform and homogenous spots. More over, probe concentration before spotting could be reduced from 0.2 to 0.02 mg/ml with PCR products and from 20 to 5 µM with 70mer oligonucleotides without affecting signal intensities after hybridisation with Cy3- and Cy5-labelled targets. More interestingly, while the binding capacity of captured probes on dendrimer-activated glass surface (named dendrislides) is roughly similar to other functionalised glass slides from commercial sources, detection sensitivity was 2-fold higher than with other available DNA microarrays. This detection limit was estimated to 0.1 pM of cDNA targets. Altogether, these features make dendrimer-activated slides ideal for manufacturing cost-effective DNA arrays applicable for gene expression and detection of mutations.     

Blinded evaluation of DNA extraction and genotyping of stained Cryptosporidium on glass slides

AIMS: A blinded trial was performed on Cryptosporidium genotyping using polymerase chain reaction (PCR)/restriction fragment length polymorphism analysis of the Cryptosporidium oocyst wall protein (COWP) gene between DNA extracted from oocyst suspensions as compared with DNA from fixed and stained faecal smears on glass microscope slides. METHODS AND RESULTS: Sixty-five faecal smears on slides were stained by one of three different methods comprising 50 positives and 15 negatives as determined by the observation of Cryptosporidium oocysts by microscopy. The expected result in terms of detection and the COWP genotype detected was achieved using DNA extracted from 94% of the slides tested. CONCLUSIONS: This study shows that DNA, which can be amplified by PCR, is present in stained smears on glass microscope slides. SIGNIFICANCE AND IMPACT OF THE STUDY: The method may be useful for molecular epidemiological studies on a range of gastrointestinal pathogens where samples are collected from locations remote from the testing laboratory.     

Theory and implementation of an electronic, automated measurement system for images obtained from immunohistochemically stained slides

OBJECTIVE: To develop and implement an Internet-based, automated image measurement system for immunohistochemically stained slides including fluorescence images in online and off-line modes. STUDY DESIGN: An image analyzing system was developed that automatically measures digitized images obtained from immunohistochemically stained slides. It is divided into a common server platform and a specific image quantification system based upon DIAS (University of Jena). After registration, the user fills in an input data form and attaches images to be measured. The server periodically transfers the data to the measurement system. The measurement works on dynamic thresholding and active sampling of objects visualized by fluorescence and conventional chromogens. It includes stereologic algorithms, object quantification, syntactic structure analysis and quality assurance. RESULTS: The system has been tested for diaminobenzidene, alkaline phosphatase and fluorescence images (FITC, etc.). The reproducibility and stability of the system are > 98%. The series of successfully measured images comprises > 1,000 images in total in the online and off-line modes. CONCLUSION: An Internet-based automated image measurement system has been developed that offers worldwide access to the major requests for quantification of immunohistochemically stained slides-tissue array analysis, nuclear stains (MIB, hormones), membrane stains (CerbB2), vascularization and fluorescence in situ hybridization.     

Thermoresponsive heparin bioconjugate as novel aqueous antithrombogenic coating material

A novel thermoresponsive aqueous antithrombogenic coating material comprising a heparin bioconjugate with a six-branched, star-shaped poly(2-(dimethylaminoethyl)methacrylate) (6B-PDMAEMA), which has both thermoresponsive and cationic characters, was developed to reduce the thrombogenic potential of blood-contacting materials such as synthetic polymers or tissue-engineered tissues in cardiovascular devices. 6B-PDMAEMA with M(n) of ca. 24 kDa was designed as a prototype compound by initiator-transfer agent-terminator (iniferter)-based living radical photopolymerization from hexakis(N,N-diethyldithiocarbamylmethyl)benzene. Bioconjugation of heparin with 6B-PDMAEMA occurred as soon as both aqueous solutions were simply mixed to form particles. The particle size at 25 °C was less than several hundred nanometers in diameter under a heparin/6B-PDMAEMA mixing weight ratio of over 2.5. The particles were very stable because of the prevention of hydrolysis of 6B-PDMAEMA in its bioconjugated form. Because the lower critical solution temperature of the bioconjugate ranges from approximately 20 to 36 °C for the formation of microparticles, the coating could be done in an aqueous solution at low temperatures. The excellent adsorptivity and high durability of the coating above 37 °C was demonstrated on silicone and polyethylene films by surface chemical compositional analysis. Blood coagulation was significantly reduced on the bioconjugate-coated surfaces. Therefore, the thermoresponsive bioconjugate developed here appears to satisfy the initial requirements for a biocompatible aqueous coating material.     

An easy technique for the clearing of histochemically stained plant tissue

The sites of GUS-activity in strongly pigmented whole-mount preparations or thick handmade sections of plant material can easily be visualized by clearing it with chlorallactophenol (CLP), without affecting the ClBr-indigo precipitate.     

Template matching as a tool for annotation of tomograms of stained biological structures

In recent years, electron tomography has improved our three-dimensional (3D) insight in the structural architecture of cells and organelles. For studies that involve the 3D imaging of stained sections, manual annotation of tomographic data has been an important method to help understand the overall 3D morphology of cellular compartments. Here, we postulate that template matching can provide a tool for more objective annotation and contouring of cellular structures. Also, this technique can extract information hitherto unharvested in tomographic studies. To evaluate the performance of template matching on tomograms of stained sections, we generated several templates representing a piece of microtubule or patches of membranes of different staining-thicknesses. These templates were matched to tomograms of stained electron microscopy sections. Both microtubules and ER-Golgi membranes could be detected using this method. By matching cuboids of different thicknesses, we were able to distinguish between coated and non-coated endosomal membrane-domains. Finally, heterogeneity in staining-thickness of endosomes could be observed. Template matching can be a useful addition to existing annotation-methods, and provide additional insights in cellular architecture.     

防白蚁电缆涂料抗台湾乳白蚁蛀蚀试验

用"群体法"对1%联苯菊酯乳油防白蚁电缆涂料抗台湾乳白蚁Coptotermes formosanus Shirak蛀蚀进行测试,结果表明:只涂防蚁涂料的电缆和木块抗白蚁性能的蛀蚀等级为1级,并且其白蚁存活时间最短,分别为10.33±1.53d和8.67±1.15d,两者之间差异不显著;先涂一层防白蚁涂料,再涂一层普通涂料的电缆和木块全达到蛀蚀等级1级,白蚁存活时间分别为24.33±2.52d和21.33±2.08d,两者之间差异不显著;只涂普通涂料的电缆和木块抗白蚁蛀蚀等级分别为1级和2～3级,白蚁存活时间分别为64.00±2.65d和62.67±2.08d,两者之间的差异也不显著;未涂涂料的电缆和木块抗白蚁蛀蚀等级分别为1级和4级,该处理的电缆缸内白蚁存活时间为72.33±3.06d,而该木块第90d试验结束时白蚁仍然活动正常,两者之间的差异显著。本研究结果显示:该防白蚁电缆涂料有良好的抗台湾乳白蚁蛀蚀性能;在电缆外护套或木块表面涂了一层该防白蚁电缆涂料后加涂一层普通涂料,既能抗台湾乳白蚁蛀蚀,又能减少环境污染。     

A laboratory-scale pulper for leafy plant material

A machine is described that, makes, from 2 to 3 kg samples of leaf, a pulp comparable to that made by the large-scale equipment used in leaf protein extraction. It is therefore suitable for use in agronomic experiments on leaf protein yield.