Late enzymes of vindoline biosynthesis. Acetyl-CoA: 17-O-deacetylvindoline 17-O-acetyl-transferase

From differentiated plants of Catharanthus roseus (L.) G. Don, a specific enzyme was isolated and named acetyl-CoA : 17-O-deacetylvindoline 17-O-acetyltransferase, acting on the biosynthetic formation of the Aspidosperma type alkaloid vindoline.The enzyme shows a high selectivity towards different substrates. The acetyl-CoA-dependent transferase also catalyses the reverse reaction by hydrolysis of the 17-O-acetyl group of vindoline in the presence of free CoA. This enzyme is localized only in vindoline-containing plant parts, but was so far not detectable in cell suspension cultures of C. roseus. The enzyme allows the synthesis of labelled vindoline with high specific activity, applicable for instance as tracer for radioimmunoassays of vindoline.     

Expression of L3T4 molecules on Lyt-2+, class II-restricted antigen-specific T suppressor lymphocytes

Three bovine serum albumin-specific Lyt-2⁺ T suppressor (Ts) cell clones from CBA/J mice have been analyzed with regard to expression of L3T4 molecules. All three Ts-cell clones can be stained with monoclonal antibodies (mAb) to L3T4. Tested for the two clones restricted to recognition of E^k determinants, antigen-specific proliferation on antigen-presenting cells, but not the proliferation induced by conditioned medium can be inhibited by L314-specific mAb. In a similar way, Ts-cell cytolytic effector functions can be blocked by L3T4-specific mAb. Thus L3T4 structures seem to play a role in Ts-cell functions. Furthermore, the data support the view that L3T4 expression can be a property of class II-restricted T cells irrespective of their Lyt phenotype.     

Cell cycle associated changes in histamine release from rat basophilic leukemia cells separated by counterflow centrifugal elutriation

This study evaluated histamine release from cells at different stages of the cell cycle. Cells from the cloned rat basophilic leukemia subline (RBL-2H3) were fractionated by counterflow elutriation according to size and density. The smallest cells were predominantly in the G1 phase of the cell cycle. These cells contained the least histamine and after IgE-mediated triggering released the lowest fraction of their total histamine. In contrast cells in the S, G2, and M stages were larger, contained more histamine and released more of their histamine after activation. When G1 stage cells were recultured, there was an increase in cell size, in histamine content and histamine release. Therefore, there is heterogeneity in the capacity of cells for IgE-mediated triggering at different stages in the cell cycle, with optimal release from the more mature cells.     

Co-amplification of rRNA genes with CAD genes in N-(phosphonacetyl)-L-aspartate-resistant Syrian hamster cells.

The amplified CAD genes in N-(phosphonacetyl)-L-aspartate (PALA)-resistant Syrian hamster cells are located in an expanded chromosomal region emanating from the site of the wild-type gene at the tip of the short arm of chromosome B-9. The terminus of B-9 in PALA-sensitive cells contains a cluster of rRNA genes (i.e., a nucleolus organizer, rDNA). We have used a molecular clone containing sequences complementary to Syrian hamster 28S rRNA to investigate whether rDNA is coamplified with CAD genes in the PALA-resistant mutants. In situ hybridization of this probe to metaphase chromosomes demonstrates that rDNA and CAD genes do coamplify in two independently isolated PALA-resistant mutants. The tight linkage of CAD and rDNA genes was demonstrated by their coordinate translocation from B-9 to the end of the long arm of chromosome C-11 in one mutant. Blot hybridization studies substantiate the in situ hybridization results. Both types of analysis indicate that only one or two rDNA genes, on the average, are coamplified per CAD gene. The data are consistent with the model that unequal exchanges between rDNA genes mediate the amplification of CAD genes in the Syrian hamster mutants that were analyzed.     

Lineare Systeme mit gesteuerten statistischen Impulsquellen

Summary From the communication engineers point of view the paper deals with networks consisting of linear filters and a special sort of controlled statistical pulse generators (SIG). The SIG responds to an analog signal with a sequence of Dirac impulses, where the probability for an output impulse at a given time depends on the instantaneous value of the input signal only. In connection with linear filters, however, systems can be realized in which the whole past of the input determines the statistical structure of the output. Therefore systems consisting of linear filters and SIGs (SIG networks) become interesting as models of biological systems, because in biological information processing transformations from analog signals into pulse trains occur very often. An example concerning the application of SIG systems in behavioural sciences will be discussed in a subsequent paper. In the present paper a theoretical analysis of the SIG and SIG networks is carried out by means of Statistical Communication Theory and Theory of Stochastic Processes. It is shown that under certain conditions very complex SIG networks can be treated with Correlation Theory. For one case not satisfying these conditions a solution on the base of Markoff processes is given.

---

---

Auszug aus einer von der Fakultät für Maschinenwesen und Elektrotechnik der Technischen Hochschule München genehmigten Dissertation.Herrn Prof. Dr.-Ing. H. Marko danke ich für das Interesse und die fördernde Kritik während des Entstehens der dieser Arbeit zugrundeliegenden Dissertation. Der Deutschen Forschungsgemeinschaft ist für die finanzielle Förderung der Untersuchungen zu danken. Die numerischen Berechnungen wurden auf der Rechenanlage TR4 des Leibniz-Rechenzentrums der Bayerischen Akademie der Wissenschaften durchgeführt.     

Licht- und elektronenmikroskopische Untersuchungen über die Beeinflussung der Dynamik neurosekretorischer Zellen von Enchytraeus (Oligochaeta) durch Aktinomycin D

Zusammenfassung Die Wirkung von Aktinomycin auf die neurosekretorischen Q-und P-Zellen im Cerebralganglion von Enchytraeus wurde untersucht. Die Cytophotometrie lichtmikroskopischer Präparate von Q-Zellen ergab, daß in den ersten Stunden nach Aktinomycin-Behandlung eine deutliche Verminderung PAF-positiven Materials auftritt. Die ersten Veränderungen wurden elektronenmikroskopisch zwischen 1 und 4 Std nach Aktinomycin-Injektion beobachtet. Sie waren in beiden Zelltypen am eindeutigsten am Nucleolus. Es kommt zu einer Sonderung und räumlichen Trennung von granulärem und fibrillärem Material. Letzteres wird sehr stark vermehrt.In bezug auf Veränderungen der Strukturen des Cytoplasmas unterscheiden sich die Q-und P-Zellen besonders im Verhalten des Golgi-Apparates und der Ribosomen. Der Golgi-Apparat wird in den Q-Zellen kurze Zeit nach Applikation von Aktinomycin reduziert. In den P-Zellen persistiert er dagegen über alle beobachteten Zeitstufen hinweg. Die Ribosomen lösen sich von den Membranen in den Q-Zellen 4–8 Std nach Injektion, was in den P-Zellen nicht festzustellen ist. Diese Tatsachen führen zu der Annahme, daß das System der Proteinsynthese der P-Zellen relativ stabiler als das der Q-Zellen ist.Die in den späteren Zeitstufen beobachtete Normalisierung der Zellstrukturen läßt darauf schließen, daß die Wirkung des einmalig injizierten Aktinomycins 24 Std danach nachzulassen beginnt.

---

Light and electron microscopic studies on the influence of actinomycin D on the dynamics of neurosecretory cells of Enchytraeus (Oligochaeta)

Summary The influence of actinomycin on the neurosecretory Q and P cells of the brain of Enchytraeus was studied. Cytophotometrical measurements of Q cells in light mirocscopic preparations showed a significant decrease of PAF-positive material in the first hours after actinomycin application. At the ultrastructural level primary changes were established one to four hours after injection of actinomycin: In the nucleolus granular and fibrillar material became separated; there was a substantial increase of the fibrillar component.Concerning structural changes of the cytoplasm, Q and P cells differed especially with respect to the Golgi apparatus and the ribosomes. In the Q cells the Golgi apparatus had become greatly reduced shortly after actinomycin treatment. However, it persisted in P cells during all stages examined. Ribosomes became detached from membranes only in Q cells between 4 and 8 hours after injection.These data indicate that protein synthesis in P cells shows greater stability than in Q cells. The restitution of normal ultrastruoture during subsequent stages indicates that effects begin to subside 24 hours after a single injection.

Für technische Unterstützung danken wir Frl. B. Reymann, Frl. A. Zinßer, Frau B. Cosack und Frau E. Wolschner.     

A STRAND OF CARDIAC MUSCLE : Its Ultrastructure and the Electrophysiological Implications of Its Geometry

The structure of a small strand of rabbit heart muscle fibers (trabecula carnea), 30–80 µ in diameter, has been examined with light and electron microscopy. By establishing a correlation between the appearance of regions of close fiber contact in light and electron microscopy, the extent and distribution of regions of close apposition of fibers has been evaluated in approximately 200 µ length of a strand. The distribution of possible regions of resistive coupling between fibers has been approximated by a model system of cables. The theoretical linear electrical properties of such a system have been analyzed and the implications of the results of this analysis are discussed. Since this preparation is to be used for correlated studies of the electrical, mechanical, and cytochemical properties of cardiac muscle, a comprehensive study of the morphology of this preparation has been made. The muscle fibers in it are distinguished from those of the rabbit papillary muscle, in that they have no triads and have a kind of mitochondrion not found in papillary muscle. No evidence of a transverse tubular system was found, but junctions of cisternae of the sarcoplasmic reticulum and the sarcolemma, peripheral couplings, were present. The electrophysiological implications of the absence of transverse tubules are discussed. The cisternae of the couplings showed periodic tubular extensions toward the sarcolemma. A regularly spaced array of Z line-like material was observed, suggesting a possible mechanism for sarcomere growth.     

Strukturanalytische Untersuchungen zur Altersabhängigkeit der Sensibilität

Zusammenfassung Eine Analyse der Meßwerte von Ronge (1943) über die Reizausnutzung durch das Tastsinnes-Nervensystem der Haut zeigt im Zusammenhang mit einer vorausgegangenen Studie (Scharf und Blumenthal, 1967), daß der Reizerfolg in einer transzendenten Fläche höherer Ordnung in Abhängigkeit vom Lebensalter (oder von der Anzahl der Meissnerschen Tastkörperchen pro Hautflächeneinheit) und vom Reizdruck dargestellt werden kann. In Druckrichtung steigt diese Fläche mit zunehmendem Reizdruck nichtlinear an, in Zeitrichtung oscilliert die Fläche dagegen träge um Normwerte, die beim 20jährigen Menschen realisiert sind. Dabei werden die Altersveränderungen der histologischen Hautstruktur offenbar zur Kompensation der altersabhängigen Verminderungen der Zahl der Tastkörperchen ausgenutzt.

---

Summary An analysis of the observations on the Reizausnutzung by nerves of touch (Ronge, 1943) connected to a previous study (Scharf and Blumenthal, 1967) shows that the irritation result may be figured by a transcendental plane of higher order as a function of age (or number of Meissner's corpuscles per area skin) and irritation pressure. Along the pressure axis this non-linear plane is increasing non-linear in dependence on ascending pressure, but along the time axis the plane oscillates lazily round about the norm values which are realized in human beings of about 20 years of age. It seems that the age-dependent changes of histological skin structure are utilized to compensate the age-dependent diminution of touch corpuscle number.

---

---

Mit dankenswerter Unterstützung durch einen Forschungsauftrag des Staatssekretariates für das Hochschulwesen der DDR.

---

Numerische Rechnung: Tischrechner Mercedes Cellatron R 44 SM, Leitende Med. techn. Ass. Ruth Pieper (Anatomisches Institut Halle). Programmgesteuerter Digitalrechner ZRA 1, Math, techn. Ass. Friedegund Hüther (Institut für Numerische Mathematik, Halle).

---

Graphik: Akad. Bildhauer Hellmut Helwin.     

Radioactive labeling of proteins in cultured postimplantation mouse embryos II. Dose and time dependency

Summary The conditions for optimum incorporation of radioactive amino acids into proteins of cultured postimplantation mouse embryos were investigated under the aspect of using these proteins for two-dimensional electrophoretic separations and fluorography. The aim was to obtain highly radioactively labeled proteins under conditions as physiological as possible. Mouse embryos of Days 8, 10, and 11 of gestation were cultured in Tyrode’s solution. Incubation time and concentration of [³H (or¹⁴C)]amino acids in the culture medium were varied over a broad range. Embryos were prepared with placenta and yolk sac or without any embryonic envelopes. After culturing, the physiologic-morphologic state of the embryos was registered on the basis of several criteria. The radioactivity taken up by the total protein of each embryo was determined and calculated in disintegrations per minute per milligram protein per embryo. To approach our aim, embryos of different developmental stages had to be cultured under different conditions. A good compromise for Day-8, Day-10, and Day-11 embryos was: embryos prepared with yolk sac (opened) and placenta, 150 μCi radioactive amino acids added per milliliter medium, incubation for 4 to 5 h. For maximum labeling of proteins it is advisable to culture Day-10 embryos without embryonic envelopes under particular conditions. This work was supported by grants from the Deutsche Forschungsgemeinschaft awarded to the project K1 237/3-2 (Systematic analysis of cell proteins).