Genetic polymorphism in normal human fibroblasts as analyzed by two-dimensional polyacrylamide gel electrophoresis.

Two dimensional gel electrophoresis has been used to measure the degree of genetic polymorphism among the proteins of normal human fibroblasts. Autoradiographic analysis of the gel protein profiles from radioactively labeled cells allowed comparison of as many as 300 discrete polypeptides at a time. In addition, a newly developed technique for double label autoradiography was used to increase the sensitivity of the system for detection of small differences in the protein profiles of different cell lines. Only about 1.2% of the proteins of different cell lines were found to differ in their electrophoretic mobility. This corresponds to an average heterozygosity of approximately 0.6%. Previous studies of genetic polymorphism using different methods of one-dimensional electrophoretic analysis have estimated the average heterozygosity of the human population at about 6.7%. Detailed mathematical analysis shows the variation of the observed from the expected number of differences to be statistically highly significant. While the reasons for this difference are not clear, the observation of low levels of genetic polymorphism on two-dimensional gels should enhance the usefulness of this technique for detection of altered proteins in inherited disease.     

Late Devonian benthic marine communities of the central Appalachian Allegheny Front

Four benthic marine communities occur in the clastic facies of the prograding Upper Frasnian-Lower Famennian (Upper Devonian) Foreknobs Formation in the Central Appalachians along the Allegheny Front in Maryland, West Virginia, and Virginia. Deep-water, rapidly prograding environments were inhabited by the Ambocoelia-Chonetes Community, dominated by an epifauna of unattached brachiopods. Offshore bar environments were inhabited by the Cyrtospirifer-Camarotoechia Community, exhibiting adaptations to shallow-water, high-energy conditions and probably lowered salinities. Shallow-water, sublittoral environments were inhabited by the Atrypa-Cypricardella Community, a community in which existed a variety of life habitats and a diverse epifaunal and infaunal association of brachiopods and bivalve molluscs. The Leptodesma-Tylothyris Community flourished in nearshore bar-protected environments in the southern region of the study area, whereas in the north the Cyrtospirifer-Camarotoechia Community inhabited nearshore environments in conjunction with the onshore development of a large fluviodeltaic system.     

Fatty Acid and Chlorophyll Levels of Coastal Bermudagrass during the Day and during Maturation

Lipids were extracted from fresh, field-grown coastal bermudagrass (Cynodon dactylon (L.) Pers.) and the fatty acids determined by gas chromatography. Total fatty acid levels (dry weight basis) increased during the day and reached a single maximum at sunset in 2-week-old grass; whereas, in older grass, the fluctuations in fatty acid levels showed two maxima. The first maximum occurred 4 h after sunrise and the second maximum occurred at sunset. Total fatty acid levels, based on dry weight, decreased during the first 6 weeks of growth and changed very little after an additional 4 weeks' growth in bermudagrass leaf blades. Chlorophyll levels (dry weight basis) continuously decreased during the entire growth period (10 weeks). Chlorophyll alb ratios increased at sunset in 2- and 6-week-old grass, but this ratio did not change during the day in subsequent growth stages. The results of these experiments show that stages of maturity affected fatty acid fluctuations during the day as well as total fatty acid and chlorophyll levels in Coastal bermudagrass leaf blades. Chlorophyll alb ratios varied independently of fatty acid levels.     

Abscisic Acid Metabolism by a Cell-free Preparation from Echinocystis lobata Liquid Endoserum

A cell-free enzyme system capable of metabolizing abscisic acid has been obtained from Eastern Wild Cucumber (Echinocystis lobata Michx.) liquid endosperm. The reaction products were determined to be phaseic acid (PA) and dihydrophaseic acid (DPA) by co-chromatography on thin layer chromatograms as the free acids, methyl esters, and their respective oxidation or reduction products. The crude enzyme preparation was separated by centrifugation into a particulate abscisic acid (ABA)-hydroxylating activity and a soluble PA-reducing activity. The particulate ABA-hydroxylating enzyme showed a requirement for O₂ and NADPH, inhibition by CO, and high substrate specificity for (+)-ABA. Acetylation of short term incubation mixtures gave evidence for the presence of 6′-hydroxymethyl-ABA as an intermediate in PA formation. Determinations of endogenous ABA and DPA concentrations suggest that the ABA-hydroxylating and PA-reducing enzymes are extensively metabolizing ABA in the intact E. lobata seed.     

Use of activated charcoal for the removal of patulin from cider.

Penicillium urticae (NRRL 2159A) was grown in culture broth containing 1 muCi of [1-14C-A1acetate to produce [14C]patulin. [14C]patulin was purified from the broth and added to apple cider. After the patulin concentration of the cider was adjusted to 30 mug/ml with unlabeled patulin, the cider was subjected to various charcoal treatments. [14C]patulin was completely removed by shaking the cider with 20 mg of activated charcoal per ml and by eluting the cider through a 40- to 60-mesh charcoal column. Activated charcola at 5 mg/ml reduced patulin in naturally contaminated cider to nondetectable levels.     

A Cannibal in the National Museum: The Early Career of Franz Boas in America

Franz Boas spent several weeks at Fort Rupert, British Columbia, at the end of 1894, when he saw the Kwakiutl hamatsa ritual in situ for the first time. Soon after his return east Boas posed for a series of photographs in the U.S. National Museum for a diorama of the hamatsa dance. These photographs, now published for the first time, are a sharp reminder of Boas' constant (and sometimes forced) collaboration with the limited number of anthropological institutions in America at the end of the century, and of his personal difficulties in establishing himself professionally in America.     

Unexplained hepatitis following halothane.

Full clinical and laboratory details of 203 patients with postoperative jaundice were submitted to a panel of hepatologists. All patients whose jaundice may have had an identifiable cause were excluded, which left 76 patients with unexplained hepatitis following halothane anaesthesia (UHFH). Hepatitis in 95% of these cases followed multiple exposure to halothane, with repeated exposure within four weeks in 55% of cases. Twenty-nine patients were obese, 52 were aged 41-70, and 53 were women. Thirteen patients died in acute hepatic failure. Rapid onset of jaundice after anaesthesia, male sex, and obesity in either sex were poor prognostic signs. Of the clinical stigmata of hypersensitivity, only eosinophilia was impressive. The UHFH group had a much greater incidence of liver kidney microsomal (LKM) and thyroid antibodies and autoimmune complement fixation than those patients whose jaundice related to identifiable factors. Thirteen of the 19 patients with LKM antibodies also had thyroid antibodies. In six patients retested two to three years later LKM antibodies had disappeared, although thyroid antibodies persisted. Rapidly repeated exposure to halothane may cause hepatitis, but such a complication is probably rare. Possibly obese women with a tendency to organ-specific autoimmunity may be more at risk. Nevertheless, the comparative risks of rapidly repeated halothane or non-halothane anaesthesia cannot be determined from the present data. If alternative satisfactory agents are available halothane should be avoided in patients with unexplained hepatitis after previous exposure, although in three to five patients with UHFH who were re-exposed to halothane jaundice did not recur.     

Nucleotide regulation of a eukaryotic protein synthesis initiation complex;.

Formation of a ternary initiation complex containing Met-tRNAf, GTP and eukaryotic initiation factor 2, is the first step in sequential assembly of the initiation complex. The concentration of GTP required for half maximal formation of the ternary complex is 2.5 with 10(-6) M. GDP is a potent competitive inhibitor of ternary complex formation with Ki = 3.4 with 10(-7) M. The nucleotide binding site on eukaryotic initiation factor 2 demonstrates relative specificity for GDP with KD(GDP) = 3.0 with 10(-8) M; 100-fold higher concentrations of GTP than GDP are required for displacement of either [(3)H]GDP or [(3)h]gtp from the necleotide binding site. An ATP-dependent stimulation of ternary complex formation observed in partially purified initiation factor preparations is due to nucleoside diphosphate kinase (EC 2.7.4.6) which serves to remove inhibitory levels of GDP by phosphorylation with ATP. Since GTP is hydrolyzed to GDP during protein synthesis, this provides a mechanism by which the ATP:ADP ratio may regulate the rate of initiation of protein synthesis.     

Messenger RNA metabolism of animal cells: possible involvement of untranslated sequences and mRNA-associated proteins

The past several years have seen a virtual revolution in the study of eukaryotic mRNA. Among the notable recent achievements are the positive identification of mRNA precursors in HnRNA, the enumeration of the DNA sequences from which mRNA is transcribed, and the finding that mRNA in cultured cells is much more stable than was previously believed. One of most far-reaching discoveries has been the finding that mRNA in eukaryotes contains poly A. This discovery, aside from providing a powerful tool for mRNA isolation, has generated a large body of research into the properties and metabolism of poly A itself. In addition, the finding of a poly A-associated protein has given a renewed stimulus to the study of proteins associated with mRNA. This review is devoted to a discussion of these and related achievements, and some of their implications