Analysis of human class I antigens by two-dimensional gel electrophoresis

Class I antigens were isolated by immunoprecipitation from cell extracts prepared from mitogenically stimulated and internally radiolabeled peripheral blood lymphocytes (PLBs). The precipitating antibodies used are monomorphic and recognize a determinant on the heavy chain of HLA-A, B, C antigens regardless of their allelic specificities when complexed with ₂m, or determinants on ₂m itself. Comparison of class I molecules isolated from 25 different homozygous typing cels (HTC) and analyzed by two-dimensional (2-D) gel electrophoresis allowed the identification of those HLA-A,13 locus specificities most common in the European Caucasoid population. Class I antigens isolated from HTC that are HLA identical are biochemically indistinguishable also. Evidence was obtained for the expression of additional class I antigens besides the HLA-A, B, C locus products: for some haplotypes, up to six class I genes may be active in mitogenically activated PBLs. No differences in molecular weight and isoelectric point of the class I heavy chains were observed between the antigens recognized by W6/32, the anti-heavy chain reagent, and anti- ₂m reagents. The nature of the mitogenic stimulus, i. e., pokeweed mitogen or phytohemagglutinin, was irrelevant with respect to the class I antigens isolated by this method. Using the HTCs as reference, a panel of HLA-B27 positive heterozygous cells was analyzed. Two types of HLA-B27 antigens, distinct by CML typing were represented. These two forms differed also in their biochemical properties. In addition, we obtained evidence for the existence of an A2 variant. This finding was likewise confirmed by CML typing.     

Mechanism of depsipeptide formation catalyzed by enniatin synthetase

Covalently bound intermediates of enniatin B synthesis could be isolated from enniatin synthetase by treatment with performic acid. By comparison with products of mild alkaline cleavage of authentic enniatin B they could be identified as the dipeptide D-2-hydroxyisovaleryl-N-methylvaline and the corresponding tetrapeptide. Synthesis of enniatins apparently proceeds via condensation of dipeptides. This was confirmed by the use of the substrate analogue isovaleric acid, which has shown to be a strong inhibitor for enniatin synthesis by formation of N-isovaleryl-N-methyl valine.     

(23S)-1,23,25-Trihydroxycholecalciferol, an intestinal metabolite of 1,25-dihydroxycholecalciferol.

(23S)-23,25-Dihydroxycholecalciferol was converted into a polar metabolite in a calciferol-deficient chick kidney homogenate. The metabolite was identified as (23S)-1,23,25-trihydroxycholecalciferol by absorbance spectroscopy and mass spectrometry, and by formation of derivatives. (23S)-1,23,25-Trihydroxycholecalciferol was also observed as a 1,25-dihydroxycholecalciferol metabolite in intestinal cells isolated from 1,25-dihydroxycholecalciferol-treated rat. The trihydroxy metabolite was 50-fold less potent than 1,25-dihydroxycholecalciferol in the chick intestinal 1,25-dihydroxycholecalciferol receptor assay.     

In vitro site-directed mutagenesis with synthetic DNA oligonucleotides yields unexpected deletions and insertions at high frequency.

We have used in vitro site-directed mutagenesis with synthetic DNA oligonucleotides to introduce single nucleotide mutations in yeast mtDNA. In addition to the expected DNA alterations we also recovered with high frequency mutants with large deletions and insertions which arose through interaction with the synthetic DNA fragment. Characterization of a number of these by DNA sequence analysis has permitted reconstruction of the mutagenic events. In all cases, the DNA fragment had base paired with non-adjacent DNA sequences sometimes more than 1000 nucleotides apart from each other on the target strand. The products of such interactions cannot be avoided due to the non-stringent annealing conditions during complementary DNA strand synthesis. However, deliberate mispairing can be directed precisely, as shown by our ability to specifically delete the 1143-bp intron from the yeast mitochondrial gene coding for large ribosomal RNA with a synthetic DNA fragment consisting of the sequence of the exon borders flanking the intron.     

The action of nitrosoguanidine and other DNA-inactivating agents on Streptococcus pyogenes K56 strains with normal and reduced dark repair ability

Summary The response pattern for ultraviolet light, nitrogen mustard, and ethyl methane sulphonate of Hcr⁺ and Hcr^- strains ofStreptococcus pyogenes K 56 is similar to that observed for analogous strains ofE. coli, whereas repair-apt streptococcal strains are much more sensitive to nitrosoguanidine and mitomycin C thanE. coli. Theuvr gene(s) appear(s) to be without effect upon survival, prophage induction, and mutation to streptomycin resistance caused by nitrosoguanidine and only of little influence on repair of mitomycin C damage.     

Licht-und elektronenmikroskopische Untersuchungen an Desmidiaceen

Ohne ZusammenfassungMit 42 Textabbildungen.     

Der Bestand vonCalanus finmarchicus in der Irminger See im Juni 1955

Zusammenfassung 1. Von den drei morphologisch unterschiedenen und zuweilen als Arten voneinander getrennten Formen des CopepodenCalanus finmarchicus s. 1. tritthelgolandicus (Claus) nur im Südosten des Untersuchungsgebietes im Nordostatlantischen Wasser auf,glacialis Yashnov ist auf das Ostgrönland-Wasser jenseits der Polarfront beschränkt, kommt aber vereinzelt auch im Tiefenwasser der Irminger See vor, währendfinmarchicus (Gunnerus) s. str. über das ganze Gebiet verbreitet ist und im Irminger-See-Wasser und dem angrenzenden Mischwasser reine Bestände bildet.2. Das Vorkommen intermediärer Formen in den Grenzgebieten sowie die großen, sich teilweise überlagernden Variationsbreiten der Merkmale lassen vermuten, daß die genannten Formen genetisch nicht streng isoliert, sondern als geographische Rassen einer plastischen ArtC. finmarchicus s. l. miteinander verbunden sind.3. Der Bestand derfinmarchicus-Rasse ist in mehrere Populationen aufgeteilt, die sich in ihrer mittleren Körpergröße und dem Entwicklungsgrad unterscheiden. Ihre Verbreitungsareale entsprechen weitgehend der hydrographischen Gliederung der Wasserkörper.4. In den Gebieten diesseits der Polarfront mit großen jahreszeitlichen Schwankungen der Temperatur besteht eine negative Korrelation zwischen der Länge der ausgewachsenen und auch jüngeren Stadien einerseits, und den örtlichen Temperaturverhältnissen andererseits. Derartige Beziehungen fehlen dagegen in den Gebieten jenseits der Polarfront mit kleiner Jahresamplitude der Temperaturschwankungen. Tiere, die durch Verdriftung oder Vermischungsvorgänge unter andere Milieuverhältnisse geraten, behalten ihr ursprüngliches Längenwachstum bei.5. Dieses unterschiedliche Verhalten gegenüber der Temperatur macht es wahrscheinlich, daß beiCalanus finmarchicus das Längenwachstum zu einem sehr frühen Zeitpunkt der Entwicklung im wesentlichen determiniert wird.

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The stock ofCalanus finmarchicus in the Irminger Sea in June 1955

During the cruise of the Fisheries Research vessel Anton Dohrn in the Irminger Sea in June 1955, the morphological and physiological composition of the stock of the copepodCalanus finmarchicus (Gunnerus) sensu lato was studied. The formhelgolandicus is restricted to the Northeast Atlantic water body, the formglacialis to the East Greenland water body, whereasfinmarchicus sensu stricto is distributed throughout the whole area. In regard to the wide variation ranges of the morphological distinctive traits and to the occurrence of intermediate forms in the border districts, the forms, hitherto described as species, are regarded as geographical races. The stock ofC. finmarchicus s. str. is divided into several populations, which differ one from another in their medium size. InC. finmarchicus, the size of adults and earlier stages is negatively related to the temperature; this correlation is lacking in populations which drifted under different temperature conditions. The distribution of the forms and populations correspond to the hydrographical and planktological pattern.

     

A comparison of sperm morphology and silver nitrate staining characteristics in the domestic ferret and the black-footed ferret

Ejaculated sperm from the domestic ferret (Mustela putorius furo) and the black-footed ferret (Mustela nigripes) were compared for differences in morphological abnormalities and argentophilic protein distribution. Thawed domestic ferret sperm was also compared to fresh sperm to determine whether there were any effects on cell morphology due to cryopreservation. There were statistically significant differences between the two species of ferret in two of the categories scored. The domestic ferret had a higher frequency of cells that were bent in the midpiece and in the principal piece, and a higher frequency of headless and tailless cells when compared to the black-footed ferret. There were no statistically significant differences in cell morphology between the fresh and cryopreserved ejaculates of the domestic ferret employing a standard egg yolk cryoextender. Silver nitrate staining distribution was different between the two species in both the head and tail region.