Low-Angle Light Scattering of Deoxyribonucleic Acid

An instrument of new design has been built in order to perform lowangle light-scattering measurements to angles as low as 16°. Native deoxyribonucleic acid preparations of different molecular weights have been studied using this apparatus with a new clarification technique. The molecular weights obtained from the low-angle data have been compared with those calculated for the same samples when using the results in the 30-150° range. The two sets of data yield the same molecular weights up to values of about 6 × 10⁶. Higher molecular weights are underestimated to a variable extent when measurements in the usual angular region (30-150°) are used.     

An inexpensive bioassay aimed at the agricultural disposal of waste waters

Zusammenfassung Es wird einfache Methode beschrieben, die von S. Prát (1947) vorgeschlagen wurde. In Petri-Schalen werden Samen des Senfes (Sinapsis alba) auf Silongewebe aufgelegt und mit dem zu untersuchenden Wasser bewässert. Das Keimen der Samen, das Wachstum der Würzelchen sowie das Läangenvehältnis von Würzelchen zu dem Hypokotyl sind als Indikatoren der wasserbeschaffenheit anzusehen. Nicht nur die toxische Einwirkung, sondern auch Wasstumsförderung kann durch diese Testmethode ermittelt werden. Es wird vorgeschlagen, diese Methode besonders bei der Beurteilung des landwirtschaftlichen Zwecken dienenden Wassers und Abwasser anzuwenden, z. B. bei der Berieselung der Felder mit Abwasser, das nicht toxisch sein darf.     

Zur Kenntnis der peritubulären Zone des Dentins

Zusammenfassung Elektronenmikroskopische Replika-Präparate (Methylmethakrylat, Bedampfung mit Chrom-Beryllium) von nativen Schliffflächen von Dentin und mit HNO₃, EDTA, KOH-Glyzerin, Äthylendiamin und Inzineration bei 300° C und 1100° C behandelte Präparate zeigen, daß sich die peritubuläre Zone von dem intertubulären Dentin in der Resistenz gegen die einwirkenden Mittel unterscheidet. Durch Inzineration bei 300° C wird sie nicht angegriffen. Säure, EDTA, KOH-Glyzerin, Äthylendiamin und Inzineration bei 1100° C greifen sie verschieden intensiv an. Die peritubuläre Zone unterscheidet sich dann häufig nicht mehr von der intertubulären Grundsubstanz. Dadurch, daß beim Polieren, bei teilweiser Inzineration und manchmal infolge Extraktion eine Fibrillenstruktur in Erscheinung tritt — wahrscheinlich durch Entfernen der Fibrillen und Darstellung des Mineralgerüstes, welches die Fibrillen einbettet — gewinnen wir einen Einblick in den Aufbau der peritubulären Zone. Anschließend wird der Strukturunterschied der peritubulären Zone dadurch bestimmt, daß die Fibrillen, die das Dentingerüst bilden, sich daselbst in feinere aufspalten, einen anderen Verlauf nehmen und von einer größeren Menge von homogener Kittsubstanz umgeben werden, die infolgedessen intensiver kalzifiziert. Obwohl die peritubuläre Zone der Dentingrundsubstanz angehört und nicht der periphere Anteil des Odontoblastenfortsatzes (Frank 1956) ist, scheint dieser bei ihrer Genese eine besondere Rolle zu spielen.     

Characterization of ATP-diphosphohydrolase activities in the intima and media of the bovine aorta: evidence for a regulatory role in platelet activation in vitro.

The inner layer of the aorta contains the enzyme ATP diphosphohydrolase (ATPDase: EC 3.6.1.5) which catalyzes the sequential phosphorolysis of ATP----ADP----AMP. Two zones of the inner layer, the intima and media, were separated and both were shown to contain ATPDase activity of similar specific activity (0.08 and 0.10 U/mg protein, respectively). However, the media exhibited about 100-times more enzyme activity than the intima. Both preparations were virtually identical with respect to pH optima (7.5), migration patterns after electrophoresis under non-denaturing conditions, relative rates of ATP and ADP hydrolysis and potency to inhibit ADP-induced platelet aggregation in both human platelet-rich plasma and whole blood. The IC50 values for ADP (2 microM)-induced aggregation were 6.8 and 12.9 mU/ml in platelet-rich plasma and whole blood, respectively. Addition of ATPDase to platelets pre-aggregated with ADP resulted in a dose-dependent disaggregation in platelet-rich plasma (IC50 4.9 mU/ml), but not in whole blood. When both ATPDase (5.6-58.7 mU/ml) and ATP (0.5-10 microM) were added to platelet-rich plasma, there was an immediate dose-dependent aggregation of platelets followed by a slowly developing disaggregation. These data show that ATPDase is present in both the intima and media layers of bovine aorta and suggest a dual role for this enzyme in platelet activation. By converting ATP released from damaged cells into ADP, the enzyme could facilitate platelet aggregation at the site of vascular injury, whereas the subsequent conversion of ADP to AMP could inhibit or reverse platelet aggregation. The consequence of these activities would be to control the growth of a platelet thrombus.     

Cytological study on wheat (Triticum timopheevi Zhuk.) protoplasts

Protoplasts were obtained from tetraploid wheat (Triticum timopheevi Zhuk.) suspension culture by incubation in solution of 1 % pectinase 500, 1 % driselase and 1 % cellulase and cultivated in Schenk and Hildebrandt medium. Freshly isolated protoplasts contained dense cytoplasm and constricted organellae exhibited negative contrast of their membranes. Together with normal protoplasts huge multinucleate protoplasts were present in the population. 3 h after plating, the cytoplasm showed normal appearance, the negative contrast of membranes was not evident any longer. Cisternae of endoplasmic reticulum and Golgi apparatus were numerous. There were some vesicles and fibres on the protoplast surface. 8 d after plating, many dividing cells were found out and cell clumps arosen in this way were present in the culture. Some of the protoplasts particularly those originally multinucleate ones were upset.     

Characterization of Cytoplasmic and Nuclear Mutations Affecting Chlorophyll and Chlorophyll-Binding Proteins during Senescence in Soybean

Soybean plants (Glycine max [L.] Merr. cv Clark) carrying nuclear and cytoplasmic “stay-green” mutations, which affect senescence, were examined. Normally, the levels of chlorophyll (Chl) a and b decline during seedfill and the Chl a/b ratio decreases during late pod development in cv Clark. Plants homozygous for both the d₁ and d₂ recessive alleles, at two different nuclear loci, respectively, retained most (64%) of their Chl a and b and exhibited no change in their Chl a/b ratio. Combination of G (a dominant nuclear allele in a third locus causing only the seed coat to stay green during senescence) with d₁d₂ further inhibited the loss of Chl in the leaf. Whereas the thylakoid proteins seem to be degraded in normal Clark leaves during late pod development, they were not substantially diminished in d₁d₂ and Gd₁d₂ leaves. In plants carrying a cytoplasmic mutation, cytG, Chl declined in parallel with normal cv Clark; however, the cytG leaves had a much higher level of Chl b, and somewhat more Chl a, remaining at abscission, enough to color the leaves green. In cytG, most thylakoid proteins were degraded, but the Chl a/b-binding polypeptides of the light-harvesting complex in photosystem II (LHCII), and their associated Chl a and b molecules, were not. Thus, the combination of d₁ and d₂ causes broad preservation of the thylakoid proteins, whereas cytG appears to selectively preserve LHCII. The cytG mutation may be useful in elucidating the sequence of events involved in the degradation of LHCII proteins and their associated pigments during senescence.     

The active site of Manganese-containing superoxide dismutase fromBacillus stearothermophilus studied by1H and19F magnetic relaxation

The dependence of the magnetic relaxation rates of¹H and¹⁹F on temperature, frequency, pH and N ₃ ^- concentration, were measured in solutions of Manganese-containing superoxide dismutase ofBacillus stearothermophilus, and were compared to activity measurements, in order to obtain some information on the structure and dynamics at Mn(III) present in the active site of the enzyme. The experimental data lead us to hypothesize the presence of two binding sites in the coordination sphere of the enzyme bound Mn(III), which are accessible to water and anions and have different chemical and spectroscopic properties. NMR measurements carried out in the presence of competitive inhibitors and the pH dependence of both NMR relaxation rates suggest that F^-, N ₃ ^- and OH^- ions bind to one site, while a water molecule binds to the other one. The stability constant values of the complexes between these anions and the enzyme are reported. The influence of the anions on activity and the pH dependence of NMR parameters are discussed.Abbreviations MnSOD Manganese containing superoxide dismutase     

Use of protoplasts in the improvement of filamentous microorganisms. I. Action of N-methyl-N′-nitro-N-nitrosoguanidine on protoplasts of Streptomycetes

Summary The effects of N-methyl-N–nitro-N-nitroso-guanidine (NTG) on protoplasts of Streptomycetes are markedly different from its action on spores, showing high mutagenic activity even at concentrations having no marked effect on protoplast survival. Strain improvement, eg in chlorotetracycline-producing strains of S. aureofaciens, was most effective when protoplasts were subjected to prolonged treatment (2 h) with low concentrations of NTG (50 /ug/ml).