A METHOD FOR OBTAINING A CONTROLLED DAILY TEMPERATURE CYCLE

An apparatus is described which gives a controlled daily cycle of temperature. The hand of a 24 hr. clock is connected by a rod to the arm operating a thermostat contact and moves the contact slowly backwards and forwards. Its method of use is described and the shape of curve it produces is expressed as a mathematical formula. A method of obtaining a nearly constant relative humidity is described.     

Solid-phase synthesis of oligomers carrying several chromophore units linked by phosphodiester backbones

A method for the preparation of oligomers by linking chromophore units is described. Specifically, the synthesis of chromophore units having a protected-hydroxyl group and a phosphoramidite function is described, along with a method to link several units using solid-phase phosphite-triester protocols.     

A chemotactic method for the isolation of Actinoplanaceae

A simple chemotactic method for the isolation of Actinoplanaceae from soil is described. The method is based on a combination of the aerotactic behavior of the spores and the attraction to chloride ions. A simple isolation chamber is described. The method is simpler and less time-consuming than the current baiting techniques.     

A simplified method for the biological assessment of the quality of fresh and slightly brackish water

A simplified method for the biological assessment of the saprobic quality of surface water is described. The method has the advantage that it is unnecessery to determine accurately the species of organisms present in the sample, which must be done when existing saprobic systems are used. The method described here cannot be used when there is a great abundance of a single species and a very strong environmental disturbance must be assumed.     

Estimation of N-amino-N,N-dimethylaminoethanol, choline and their acetate esters by gas chromatography mass spectrometry

A method is described for measurement of choline, N-aminodeanol, and their acetyl esters by gas chromatography mass spectrometry. The preparation of N-aminodeanol and its isotopic variants is also described. This method allows a thorough quantitative analysis of the replacement of true with false neurotransmitter in biological preparations.     

Measurement of glycolytic rates in cell suspensions using a recording pH meter.

A method for measuring continuously glycolytic rates in cell suspensions, using a recording pH meter, is described. Under the described conditions the method is very exact, sensitive and reproducible. The method can be applied to different cells and different conditions of assay calibrating in each case the pH range, cell concentration range and the ratio of delta protons to delta lactic acid.     

A New method for designing PCR primers specific for groups of sequences and its application to plant viruses

A new method is described for identifying short regions of sequence similarity in a group of selected sequences. These regions have been used for the design of both specific and degenerate PCR primers for the detection of groups of plant viruses, but the method has wider applications. The method is an extension of the GCG programs COMPARE and DOTPLOT, so the name “dot primers” is suggested as a generic term for primers designed in this way. The method described is more direct and more efficient than current methods that use sequence alignment algorithms.     

A simplified purification method of aminoacylase I

Taking advantage that the stability of aminoacylase I increases in the presence of Co2+, a simplified method of its purification, which includes a heating step, is described. This method is easy, fast, and gives a good yield of the enzyme. The properties of aminoacylase I thus prepared are similar to those described in the literature.     

Technique for measuring picomolar amounts of bound and unbound amino acids obtained from myogenic cell cultures of skeletal muscle

Summary A procedure is described that compares the isotope dilution method of measuring picomolar amounts of amino acids obtained from cellular extracts with a direct method of analysis. Evidence is provided that shows that the direct method is at least as accurate as the isotope dilution method. In addition the direct method is as expedient and requires but a single isotope and fewer chromatograms for analysis. A procedure also is described for selecting the appropriate conditions for dansylation and for measuring the loss of dansyl amino acid due to decomposition. This research was funded through a grant from the Muscular Dystrophy Association of Canada.     

A method for cerebroventricular cannulation of the rat

A convenient technique is described for the preparation and surgical implantation of permanent cannulae into the lateral ventricles of the rat brain, as well as a method for radiographic confirmation of the location of the implanted cannula. The modified implantation method described is simple, rapid and has produced low mortality in the surgically altered animals.     

Archaeological Inference and Inductive Confirmation

The limitations of the hypothetico-deductive (H-D) method of inductive confirmation are described, and an alternate method, the hypothetico-analog (H-A) method is described in detail. The H-A method can be characterized as a modified and supplemented form of the simple H-D method, and is proposed as being more appropriate for archaeological inference. Aspects of the H-A method that are given particular attention include the establishment of boundary conditions for reference classes, plausibility considerations, multiple working hypotheses, bridging arguments, and criteria for selecting alternative hypotheses . [scientific method, archaeological inference, hypothetico-analog method, inductive confirmation]     

A rapid method for analyzing the ligation products of synthetic oligodeoxyribonucleotides

A method is described for the rapid analysis of DNA ligation products in the assembly of synthetic genes and gene fragments. The method is based on the simultaneous analysis of multiple ligation reactions where a single but different DNA oligomer is radiolabelled per ligation reaction. After each ligation the reaction mixture is electrophoresed on a denaturing, as well as a non-denaturing, polyacrylamide gel allowing one to monitor the ligation reaction products. In addition, a unique method for generating single stranded DNA sizing standards up to approximately 300 nucleotides in length is described.     

Electrophoresis, crossed immunoelectrophoresis, and isoelectric focusing in agarose gels with reduced electroendosmotic flow

An ion exchange method using QAE-Sephadex for preparation of agarose with a low electroendosmotic flow and reduced adsorption properties is described. The successful use of such agarose for the separation of highly cationic proteins is illustrated.A method for isoelectric focusing of proteins in gels made from a mixture of purified agarose and a water-soluble non-cross-linked acrylamide polymer is described. This method can be combined with immunochemical identification by electrophoresis of the separated components into antibody-containing agarose gels, also containing such a polymer of acrylamide.     

A new, rapid, method for preparation of dispersed pancreatic acini.

A new method for the preparation of pancreatic acini is described. The method is simple and much more rapid than previously described techniques, the time required for preparation of pancreatic acini being 20 min from removal of the pancreas. Acini prepared with this method perform in a superior manner when stimulated by either caerulein or secretin. Thus this new technique would be ideal for use in binding and secretion studies.     

Isolation of high quality RNA from bilberry (Vaccinium myrtillus L.) fruit

A simple and efficient method is described for isolating high quality RNA from bilberry fruit. The procedure is based on the use of hexadecyltrimethyl ammonium bromide (CTAB), polyvinylpyrrolidone (PVP), and β-mercaptoethanol in an extraction buffer in order to eliminate the polysaccharides and prevent the oxidation of phenolic compounds. This method is a modification of the one described for pine trees, and yields high-quality RNA suitable for cDNA based methodologies. This method is applicable for a variety of plant tissues.     

Corneal endothelium: a modified method for cultivation

A modified method for establishing cultures of rabbit corneal cells is described. The new technique utilized a Lucite disc in combination with a Tygon ring for growth of pure cell cultures and was compared with an explant method for growing cells. Each method provided adequate cell cultures for biochemical or ultrastructure studies of rabbit corneal cells, but the ring and disc method described here allowed the isolation of specific cell types without the interference of stromal cell contamination.     

Corneal endothelium: A modified method for cultivation

Summary A modified method for establishing cultures of rabbit corneal cells is described. The new technique utilized a Lucite disc in combination with a Tygon ring for growth of pure cell cultures and was compared with an explant method for growing cells. Each method provided adequate cell cultures for biochemical or ultrastructure studies of rabbit corneal cells, but the ring and disc method described here allowed the isolation of specific cell types without the interference of stromal cell contamination.     

Enzymatic synthesis of [beta-32P]ADP using adenylate kinase and [gamma-32P]ATP

An enzymatic method for the synthesis of [beta-32P]ADP from [gamma-32P]ATP is described. This substrate is required for the assay of ADPase and is not commercially available. The method described results in a preparation of [beta-32P]ADP of high purity with a yield of approximately 40% the theoretical obtainable.     

The Use of Electron Tomography for Structural Analysis of Disordered Protein Arrays

A method based on Fourier transforms is described for obtaining a 3-D reconstruction from a paracrystalline object with static disorder. The method is derived from the standard methods used in 3-D reconstruction of 2-D crystals except that all of the Fourier coefficients are used and not just the sampled data from the periodic lattice. Thus, not only is the spatially ordered part of the structure visualized in 3-D, but also the spatially disordered part. Application of the method to 3-D reconstructions of insect flight muscle is described as well as prospects for extension of the method to radiation-sensitive specimens.     

Purification of alpha2-macroglobulin and the construction of immunogenic alpha2-macroglobulin-peptide complexes for use as cancer vaccines

A simple method for purification of immunogenic alpha2-macroglobulin from murine serum is described. Also described is a method for creating alpha2-macroglobulin-antigen complexes using purified alpha2-macroglobulin and peptides derived from tumor cell lysates. The application of such alpha2-macroglobulin-antigen complexes for eliciting cancer immunity is discussed.