Discriminant analysis on cells from developing squamous cancer of the respiratory tract

Cytologic preparations made from the tracheobronchial tree taken by the Schreiber catheter have been scanned by three color microphotometry. The digitized cell images were processed by the analytical cytodiagnostic programs of the TICAS system. Cells were sorted into two control groups and five groups of increasing atypia ranging from normal epithelium to invasive squamous cell carcinoma. Standard statistical tests, including Wilk's Lambda, Rao's V, and the Kruskal-Wallis tests are performed on these subsets of cell image features. This study demonstrates that discriminant analyses permit differentiation between normal cells and those from marked atypia or carcinoma and that the classification achieves a high degree of agreement with visual assignment.     

Homology between ricin and Ricinus communie agglutinin: Amino terminal sequence analysis and protein synthesis inhibition studies

The existence of three forms of ricin and two forms of the Ricinus communis agglutinin (RCA) was established using cation exchange chromatography, isoelectric focusing, and polyacrylamide gel electrophoresis. The preparation of the RCA we obtained was 60–75 times more potent than ricin in the agglutination of erythrocytes, but was about 4% as effective as an inhibitor of cell-free protein synthesis. When reduced with 2-mercaptoethanol, the RCA was activated 3000-fold as an inhibitor of cell-free protein synthesis, whereas ricin was activated about 600-fold by the same treatment. A mixture of the RCA A chains was about one-fifth as effective as the ricin A chain in the inhibition of cell-free protein synthesis. The purified polypeptide subunits of the castor bean lectins were subjected to automated Edman degradation. The sequence for 17 of the first 19 residues of the agglutinin A chain was determined. The first seven residues of the ricin A chain were determined and they are identical with those of the RCA A chain. Nineteen turns of Edman degradation on the RCA B chain resulted in the identification of 18 amino acids. The sequence determined for the first 17 residues of the ricin B chain was identical with that of the RCA B chain. It is likely that the identity of the ricin/RCA A and B chain sequences extends further along the polypeptide chains than the sequences we have determined. The similar structural and catalytic potentials of the RCA and ricin suggest that they bear a precursor-product relationship.     

Control of acetylcholine receptor mobility and distribution in cultured muscle membranes. A fluorescence study

The molecular control of the distribution and motion of acetylcholine receptors in the plasma membrane of developing rat myotubes in primary cell culture was investigated by fluorescence techniques. Acetylcholine receptors were marked with tetramethylrhodamine-labeled α-bungarotoxin and lateral molecular motion in the membrane was measured by the fluorescence photobleaching recovery technique. Three types of experiments are discussed: (I) The effect of enzymatic cleavages, drugs, cross-linkers, and physiological alterations on the lateral motion of acetylcholine receptors and on the characteristic distribution of acetylcholine receptors into patch and diffuse areas. (II) Observation of the distribution and/or motion of fluorescence-labeled concanavalin A receptors, lipid probes, cell surface protein, and stained cholinesterase in acetylcholine receptor patch and diffuse areas. (III) The effect of a protein synthesis inhibitor and electrical stimulation on membrane incorporation of new acetylcholine receptors.Some of the main conclusions are: (a) acetylcholine receptor lateral motion is inhibited by concanavalin A plant lectin and by anti-α-bungarotoxin antibody, but marginally enhanced by treatment with a local anesthetic; (b) patches are stabilized by an immobile cellular structure consisting of molecules other than the acetylcholine receptors themselves; (c) this structure is highly selective for acetylcholine receptors and not for other cell membrane components; (d) acetylcholine receptor patch integrity and diffuse area motion are independent of direct metabolic energy requirements and are sensitive to electrical excitation of myotube; (e) lipid molecules can move laterally in both acetylcholine receptor patches and diffuse areas; and (f) acetylcholine receptor lateral motion in diffuse areas and immobility in patch areas are not altered by specific agents which are known to affect extrinsic cell surface proteins, or cytoplasmic microfilaments and microtubules.     

Assignment of the gene for dipeptidase 2 to Mus musculus chromosome 18 by somatic cell hybridization

Evidence is presented for the assignment of the gene for dipeptidase 2 to Mus musculus chromosome 18 by synteny testing and karyotypic analysis of Chinese hamster × mouse somatic cell hybrid clones. DIP-2 and chromosome 18 were expressed concordantly in 24/24 clones examined (ten primary clones and 14 secondary clones). Synteny testing indicated that DIP-2 was not expressed concordantly with the expression of any marker enzymes.This work was supported by NIH grant USPHS GM 09966.     

Assignment of the gene for glyoxylase I to mouse chromosome 17 by somatic cell genetics

Evidence is presented for the assignment of the gene for glyoxylase I to mouse chromosome 17 using mouse × Chinese hamster somatic cell hybrids. GLO I was not expressed concordantly with any known marker enzymes which represented 11 linkage groups. The presence of chromosome 17 and expression of GLO I were concordant in 31/31 clones. GLO I is thus linked to the H-2 histocompatibility locus in the mouse.     

The wavelength dependence of the turbidity of solutions of macromolecules

The experimental parameter describing the wavelength dependence of the turbidity of solutions of macromolecules is usually the negative slope of the graph of the logarithm of optical density vs the logarithm of the in vacuo wavelength, λ₀. Such slopes are the apparent exponents of 1/λ₀ in the turbidity equation. Their values depend upon the way the destructive interference of the scattered light, the refractive index increment, and the solvent refractive index change with wavelength. In this study, expressions for the wavelength exponents for isotropic rods, spheres, and random coils have been obtained and evaluated by representing the intraparticle interference functions, Q(λ₀), with series in even powers of 1/λ₀ and the refractive properties with Cauchy relations. Comparisons of calculated and observed exponents at wavelengths in the visible spectral region for aqueous solutions of four viruses have been made: for R17, T7, and PM2 bacteriophage, the exponents are greater than four; whereas for tobacco mosaic virus, they are less than four. The application of the turbidity relations to determine the size and molecular weight of biological macromolecules is discussed.     

A general procedure for the preparation of labeled l-ascorbic acid from labeled d-glucose

A simple, three-step conversion of 1,2-O-isopropylidene-α-d-glucofuranose into l-ascorbic acid, originally described by Bakke and Theander, was used to prepare l-[4-¹⁴C]ascorbic acid from milligram amounts of d-[3-¹⁴C]glucopyranose in 28% radioisotopic yield. In addition, l-[6-¹⁴C]- and l-[U-¹⁴C]-ascorbic acid were prepared from d-[1-¹⁴C]- and d-[U-¹⁴C]-glucopyranose, respectively. The procedure is useful for the synthesis of l-ascorbic acid bearing isotopic hydrogen, carbon, or oxygen atoms at specific positions, subject only to the availability of starting material.