Lebendbeobachtungen zur Struktur des Golgifeldes bei Zellen eines transplantablen Plasmocytoms in vitro

Zusammenfassung Zellen des aus unreifen Plasmoblasten bestehenden, transplantablen KG 13-Plasmocytoms des Goldhamsters zeigen bei phasenoptischer Lebendbeobachtung in vitro ein besonders großes, stark strukturiertes Golgifeld. Dieses wird von einem Saum heller Pinocytosetröpfchen gegen die Speichergranula abgesetzt. Die Form des Golgifeldes, die Gesetzmäßigkeit seiner Veränderungen, die Spezifität seiner Strukturen und seine Beziehung zum Zellkern werden beschrieben. Pinocytosebläschen wandern bei dieser Zellart im gesamten Verlauf sichtbar von der Zellmembran zwischen den Speichergranula hindurch zur Randzone des Golgifeldes und lösen sich dort auf. Die Granula des Cytoplasmas entstehen offenbar im Golgifeld und werden in seiner Randzone erstmalig als solche erkennbar, um ins Cytoplasma zu wandern.

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Summary The KG 13 plasmocytoma from the Syrian golden hamster consists of immature Plasmoblasts. Living cells of this plasmocytoma observed by phase contrast microscopy in the hanging drop in vitro have a particularly large Golgi field surrounded by a border of light pinocytose bubbles. The cytoplasma is filled with storage granules. The form and the modifications of the Golgi field are described as well as his structure and his relations to the nucleus. Pinocytose bubbles reach the border of the Golgi field after crossing granules and disappeare. The granules are evidently formed in the Golgi field and become visible in its periphery.

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Die Arbeit wurde mit Unterstützung durch die Deutsche Forschungsgemeinschaft durchgeführt.     

Coaction of blue/ultraviolet-A light and light absorbed by phytochrome in controlling growth of pine (Pinus sylestris L.) seedlings

Photomorphogenesis is a conspicuous feature in conifers. In the case of the shade-intolerant Scots pine (Pinus sylvestris L.), control of stem growth by light is well expressed at the seedling stage and can readily be studied. The present data show that hypocotyl growth is controlled by the far-red-absorbing form of phytochrome (Pfr). However, the Scots pine seedling requires blue or ultraviolet (UV-A) light to become fully responsive to Pfr. Blue/UV-A light has no direct effect on hypocotyl growth and its action appears to be limited to establishing the responsiveness of the seedling to Pfr. This type of coaction between phytochrome and blue/UV-A light has been observed previously in a number of angiosperm seedlings. With regard to the high irradiance reaction of phytochrome in long-term far-red light the pine seedling deviates totally from what has been observed in etiolated angiosperms since continuous far-red light has no effect on stem growth.Abbreviations B light of wavelength between 500 and 400 nm - FR standard far-red light - HIR high irradiance reaction of phytochrome - R high-fluence-rate red light (_R = 0.8) - RG9-light long-wavelength far-red light defined by the properties of the Schott RG9 glass filter (_RG₉<0.01) - = Pfr/Ptot wavelength-dependent photoequilibrium of the phytochrome system (far-red-absorbing form of phytochrome/total phytochrome) - UV-A near ultraviolet light of wavelength between 400 and 320 nm - W white light Research supported by a grant from the Deutsche Forschungsgemeinschaft (Schwerpunkt Physiologie der Bäume).     

Alveolar slope and dead space of He and SF6 in dogs: comparison of airway and venous loading

Series (Fowler) dead space (VD) and slope of the alveolar plateau of two inert gases (He and SF6) with similar blood-gas partition coefficients (approximately 0.01) but different diffusivities were analyzed in 10 anesthetized paralyzed mechanically ventilated dogs (mean body wt 20 kg). Single-breath constant-flow expirograms were simultaneously recorded in two conditions: 1) after equilibration of lung gas with the inert gases at tracer concentrations [airway loading (AL)] and 2) during steady-state elimination of the inert gases continuously introduced into venous blood by a membrane oxygenator and partial arteriovenous bypass [venous loading (VL)]. VD was consistently larger for SF6 than for He, but there was no difference between AL and VL. The relative alveolar slope, defined as increment of partial pressure per increment of expired volume and normalized to mixed expired-inspired partial pressure difference, was larger by a factor of two in VL than in AL for both He and SF6. The He-to-SF6 ratio of relative alveolar slope was generally smaller than unity in both VL and AL. Whereas unequal ventilation-volume distribution combined with sequential emptying of parallel lung regions appears to be responsible for the sloping alveolar plateau during AL, the steeper slope during VL is attributed to the combined effects of continuing gas exchange and ventilation-perfusion inequality coupled with sequential emptying. The differences between He and SF6 point at the contributing role of diffusion-dependent mechanisms in intrapulmonary gas mixing.     

The E1 replication protein of bovine papillomavirus type 1 contains an extended nuclear localization signal that includes a p34cdc2 phosphorylation site.

Bovine papillomavirus (BPV) DNA replication occurs in the nucleus of infected cells. Most enzymatic activities are carried out by host cell proteins, with the viral E1 and E2 proteins required for the assembly of an initiation complex at the replication origin. In latently infected cells, viral DNA replication occurs in synchrony with the host cell chromosomes, maintaining a constant average copy number of BPV genomes per infected cell. By analyzing a series of mutants of the amino-terminal region of the E1 protein, we have identified the signal for transport of this protein to the cell nucleus. The E1 nuclear transport motif is highly conserved in the animal and human papillomaviruses and is encoded in a similar region in the related E1 genes. The signal is extended relative to the simple nuclear localization signals and contains two short amino acid sequences which contribute to nuclear transport, located between amino acids 85 and 108 of the BPV-1 E1 protein. Mutations in either basic region reduce nuclear transport of E1 protein and interfere with viral DNA replication. Mutations in both sequences simultaneously prevent any observable accumulation of the protein and reduce replication in transient assays to barely detectable levels. Surprisingly, these mutations had no effect on the ability of viral genomes to morphologically transform cells, although the plasmid DNA in the transformed cells was maintained at a very low copy number. Between these two basic amino acid blocks in the nuclear transport signal, at threonine 102, is a putative site for phosphorylation by the cell cycle regulated kinase p34cdc2. Utilizing an E1 protein purified from either a baculovirus vector system or Escherichia coli, we have shown that the E1 protein is a substrate for this kinase. An E1 gene mutant at threonine 102 encodes for a protein which is no longer a substrate for the p34cdc2 kinase. Mutation of this threonine to isoleucine had no observable effect on either nuclear localization of E1 or DNA replication of the intact viral genome.     

Die histologischen Vorgänge während der lichtabhängigen Schliessung und Öffnung des Plumulahakens bei den Keimlingen vonLactuca sativa L.

Summary In a preceding paper the following facts have been shown (Mohr undNoblé 1960): Dark grown seedlings ofLactuca sativa L. var.Grand Rapids (tip burn resistant strain) do not form an appreciable plumular hook. But a normal plumular hook, as is formed for instance byPhaseolus vulgaris in complete darkness, can be induced by red light. This formation of the plumular hook is controlled by the phytochrome system. — The hook which has been closed by red radiation can be reopened by light. In this respect only the high energy reaction of photomorphogenesis is effective. This reaction system shows peaks of action in the blue and in the far-red part of the visible spectrum (cf.Mohr 1962).The present paper deals with the investigation of the anatomical and cellular changes which lead to hook formation and reopening. The movements of the hook always take place in the cotyledonary plane. Bending of the hook accurs because the cells on the potentially outer side of the future hook increase their rate of elongation, unbending of the hook is due to a rapid increase in rate of elongation of the cells on the inner side of the closed hook. Cell dividions are apparently not involved either in the bending or in the unbending of the hook. It is concluded, that the apical part of the hypocotyl is a highly complex organ the potentialities of which can be described only with the help of physiological experimentation. The conventional anatomical or cytological approaches are not adequate to describe in detail the pattern of differentiation which must be present in the apical part of the hypocotyl according to the reactions which can be induced by light.

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Als Plumulahaken (=plumular hook) bezeichnen wir die auffällige Einkrümmungszone im Apikalbereich einer Keimpflanze. Ein knapper und klarer Begriff hat sich offenbar in der deutschen Literatur nicht durchgesetzt. AuchTroll (1959) spricht nur allgemein von einer apikalen Krümmung des Keimsprosses. Bei dem Plumulahaken derLactuca-Keimlinge handelt es sich um eine Einkrümmung des apikalen Hypokotylbereichs.     

Eine mit Xenonbögen ausgerüstete Interferenzfilter-Monochromatoranlage für kurzwellige sichtbare und Langwellige ultraviolette Strahlung

Summary An interference filter monochromator system for biological purposes has been described. The system contains two monochromator units. It operates with xenon arcs (Osram XBO 501) as sources of radiation and is therefore especially adapted for experimental work in the blue-violet range of the visible spectrum and in the near ultraviolet (until about 350 m). All lenses are made of quartz. A circle of 8 cm diameter is homogenously irradiated. The irradiances which can be presently obtained within this circle with our single or double interference filters are rather high (1900 and 630 ergs/cm²·sec respectively at 355 m, 9000 and 1700 ergs/cm²·sec respectively around 450 m). Together with the monochromator system which we described recently (Mohr undSchoser 1959) and which operates with incandescent projection lamps this xenon arc system enables biological investigations with highly purified monochromatic radiation in the spectral range from 350 to 800 m.

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Biochemical bone marrow markers and their significance in postmenopausal osteoporosis--a new method in the diagnosis of osteoporosis?]

This study analyzes the qualification of biochemical markers in the diagnosis of osteoporosis and evaluates the potential of a multiparametric classification of premenopausal and non-osteoporotic as well as osteoporotic postmenopausal women, which is based on biochemical marker profiles. For this evaluation data of 29 women in the age between 28-74 years were used. The classification of osteoporosis was done by the trabecular density of the lumbar spine using qCT-measurements. The biochemical markers of formation and resorption AP, bAP, OC, ucOC, PICP, PYD, DPD, NTX, BSP and vitamin K were analyzed on day 1 and 42 in all patients. For vitamin K we found significant distribution differences between non-osteoporotic and osteoporotic women (p<0.005). The crosslinks PYD and DPD showed weakly significant differences. All other parameters exhibited non-significant results. Vitamin K acted with a sensitivity of 64% and a specificity of 82%. The used multiparameter classification process improved sensitivity and specificity considerably. The parameter profiles of OC/PYD, vitamin K/PYD and vitamin K/bAP revealed the highest sensitivities with specificities of more than 82%.     

Seven New Mutations in hMSH2, an HNPCC Gene, Identified by Denaturing Gradient-Gel Electrophoresis

Hereditary nonpolyposis colorectal cancer (HNPCC) is a relatively common autosomal dominant cancer-susceptibility condition. The recent isolation of the DNA mismatch repair genes (hMSH2, hMLH1, hPMS1, and hPMS2) responsible for HNPCC has allowed the search for germ-line mutations in affected individuals. In this study we used denaturing gradient-gel electrophoresis to screen for mutations in the hMSH2 gene. Analysis of all the 16 exons of hMSH2, in 34 unrelated HNPCC kindreds, has revealed seven novel pathogenic germ-line mutations resulting in stop codons either directly or through frameshifts. Additionally, nucleotide substitutions giving rise to one missense, two silent, and one useful polymorphism have been identified. The proportion of families in which hMSH2 mutations were found is 21%. Although the spectrum of mutations spread at the hMSH2 gene among HNPCC patients appears extremely heterogeneous, we were not able to establish any correlation between the site of the individual mutations and the corresponding tumor spectrum. Our results indicate that, given the genomic size and organization of the hMSH2 gene and the heterogeneity of its mutation spectrum, a rapid and efficient mutation detection procedure is necessary for routine molecular diagnosis and presymptomatic detection of the disease in a clinical setup.     

Coaction of three factors controlling chlorophyll and anthocyanin synthesis

In a three-factor analysis the rate of chlorophyll a (Chl) accumulation in excised mustard cotyledons was studied as a function of kinetin, light (operating through phytochrome, P _fr) and an excision factor. It was found that the three factors operate additively provided that the P _fr level is high enough. When the P _fr level is below approximately 1 per cent (<0.01) the effectiveness of the excision factor decreases while the effect of kinetin remains additive. The observed additivity is explained by a model where the three factors operate independently through a common intermediate (presumably 5-aminolevulinate) in the biosynthetic chain leading to Chl. With regard to the coaction of the excision factor and phytochrome it is concluded that the production of the excision factor requires the operation of phytochrome (even though saturated at a low P _fr level) while the action of the excision factor is independent of phytochrome. This conclusion was confirmed by experiments in which the rate of light-mediated anthocyanin synthesis was measured in excised mustard cotyledons. The effect of excision in the case of anthocyanin formation differs kinetically from the effect of excision on Chl formation.Abbreviations Chl chlorophyll(ide) a - P _fr far-red absorbing form of phytochrome - P _fr/P _tot ratio at photoequilibrium - RL red light - FR far-red light - GL green light - RG9 light long wavelength far-red light - WL white light