Lectins as probes of chromatin structure. Binding of concanavalin A to purified rat liver chromatin

Concanavalin A (Con A) binds specifically to rat liver chromatin. The extent of binding is directly proportional to both chromatin and concanavalin A concentration. It is reversible and inhibited by specific sugars for which concanavalin A has a binding site. Scatchard analysis reveals the presence of one type of Con A-binding site, with an apparent dissociation constant of 3 X 10(-7) M. A maximum of 10 pmol of Con A binds to 10 microgram of chromatin, indicating an average of one binding site/1400 base pairs of DNA. To identify the polypeptide chains which contain Con A-binding sites, chromosomal proteins were separated by electrophoresis on polyacrylamide gels containing sodium dodecyl sulfate. Con A receptors were localized by incubating the gel in 125I-Con A and subsequent autoradiography. Three major polypeptide bands which bind Con A were identified among the nonhistone chromosomal proteins. The apparent molecular weights of these glycoproteins are 135,000, 125,000, and 69,000. We suggest that lectins may serve as probes for the study of the organization of specific components in chromatin.     

Human mesotheliomas contain the simian virus-40 regulatory region and large tumor antigen DNA sequences

BACKGROUND: A cohort (20%) of patients with mesothelioma will not have an exposure to asbestos. Recently, a DNA tumor virus (simian virus 40) has been shown to cause hamster mesotheliomas; we previously described simian virus 40-like DNA amino terminus sequences in 29 of 48 mesotheliomas. We analyzed an additional 42 mesotheliomas to determine (1) whether our initial observations were durable and (2) the extent to which the simian virus 40 genome is present in mesotheliomas. METHODS: Genomic DNA was extracted from snap frozen mesothelioma tumor samples and from the simian virus 40-induced hamster mesothelioma tumor H9A. Polymerase chain reaction primers were used to amplify various simian virus 40 large T-antigen regions including a 105-base pair amino terminus fragment, a 281-base pair carboxyl terminus fragment, and a 310-base pair fragment of the enhancer promoter region. Endonuclease digestions and Southern blotting were used to verify the expected product. RESULTS: Thirty of the 42 (71%) samples amplified T-antigen amino sequences, and specificity was verified by Southern hybridization. Sixteen of 42 samples (38%) amplified the appropriate size fragment for the carboxyl terminus, and digestion with BsaB1 matched that of H9A. Twenty-two of 42 samples (52%) amplified simian virus 40 regulatory sequences and Fok1 digestion matched that of the hamster control tumor. Sequence analysis (4 patients) revealed 100% homology with the regulatory region of simian virus 40 strain 776. CONCLUSIONS: These data suggest an association between the simian virus 40 virus and human mesothelioma that could be exploited for diagnostic/therapeutic options including early detection and potential vaccination strategies.     

Co-oligopeptides of glycine and aromatic amino acids with variable distance between the aromatic residues. IV. Spectroscopic properties of tryptophan-containing peptides in a cyclohexane phase

L-Tryptophan, L-tryptophanylglycine, glycyl-L-tryptophan, glycyl-L-tryptophanylglycine and glycyl-L-tryptophanylglycylglycyl-L-tryptophanylglycine have been transferred from an aqueous solution (generally 0.1 M NaOH) to cyclohexane, using the quaternary ammonium salt trioctylmethyl ammonium chloride (NR+4Cl-, soluble in cyclohexane but not in water) as the transporting agent. The spectroscopic properties of L-tryptophan and tryptophan-containing peptides have been studied in the cyclohexane phase. With respect to the aqueous solutions, ultraviolet absorption spectra are characterized by a considerable red shift of the absorption maxima and by a hypochromicity of up to 10%. Fluorescence spectra generally show emission maxima which are characteristic of polar environments, accompanied by a significant enhancement of the quantum yield. CD spectra have also been investigated for all peptides and compared with those for aqueous systems reported in preceding publications. All these spectral changes cannot be attributed solely to the cyclohexane solvent effect. It is suggested that these anomalous spectral properties of the tryptophan-containing compounds in the cyclohexane-NR+4 solution are due to the influence the electrostatic field of the ion pair has on the indole chromophore. The possible implications of this finding for the spectroscopic properties of aromatic residues buried in the polar interior of proteins are discussed.     

'Bálint's syndrome' and associated visuospatial disorders

There are several objections to the need for the designation 'Bálint's syndrome'. The full syndrome is generally associated with a wide variety of behavioural disturbances outside Bálint's original triad that may confound the interpretation of their supposed defect. It embeds itself in the hemineglect syndrome, violating the principle of autonomy for a syndrome. Individual components of the triad, particularly simultanagnosia, may represent relatively broad categories that subsume other combinations of defects of widely different degrees of severity. The diagnosis fails to predict a consistent site of anatomical impairment, and the main proposed mechanism is not sound. Our experience with a registry of patients with focal brain lesions and well-documented behavioural impairments (Palca, 1990), now numbering over 1500 individuals, tends to support these generalizations.