Combination of arsenic trioxide and BCNU synergistically triggers redox-mediated autophagic cell death in human solid tumors

Arsenic trioxide (As₂O₃) is an effective treatment for relapsed or refractory acute promyelocytic leukemia (APL). After the discovery of As₂O₃ as a promising treatment for APL, several studies investigated the use of As₂O₃ as a single agent in the treatment of solid tumors; however, its therapeutic efficacy is limited. Thus, the systematic study of the combination of As₂O₃ with other clinically used chemotherapeutic drugs to improve its therapeutic efficacy in treating human solid tumors is merited. In this study, we demonstrate for the first time, using isobologram analysis, that As₂O₃ exhibits a synergistic interaction with N,N′-bis(2-chloroethyl)-N-nitrosourea (BCNU). The synergistic augmentation of the cytotoxicity of As₂O₃ with BCNU is in part through the autophagic cell death machinery in human solid tumor cells. As₂O₃ and BCNU in combination produce enhanced cytotoxicity via the depletion of reduced glutathione (GSH) and augmentation of reaction oxygen species (ROS) production. Further analysis indicated that the extension of GSH depletion by this combined regimen occurs through the inhibition of the catalytic activity of glutathione reductase. Blocking ROS production with antioxidants or ROS scavengers effectively inhibits cell death and autophagy formation, indicating that redox-mediated autophagic cell death involves the synergism of As₂O₃ with BCNU. Taken together, this is the first evidence that BCNU could help to extend the therapeutic spectrum of As₂O₃. These findings will be useful in designing future clinical trials of combination chemotherapy with As₂O₃ and BCNU, with the potential for broad use against a variety of solid tumors.     

Elektronenmikroskopische Untersuchungen an spezifischen Organellen von Endothelzellen des Frosches (Rana temporaria)

Zusammenfassung Endothelzellen der Wirbeltiere enthalten spezifische Organellen, deren Funktion unbekannt ist. Diese Organellen werden beim Frosch (Rana temporaria) nach unterschiedlichen Fixierungen elektronenmikroskopisch untersucht. Die Organellen sind walzenförmig mit mannigfachen Abweichungen, bis zu 2 lang und 0,1 bis 0,5 dick. Ihre oft unterbrochene Außenmembran ist dicker als zytoplasmatische Membranen. Das Innere der Organellen besteht aus Tubuli, die in eine elektronendichte Matrix eingebettet sind. Die Dichte dieser Matrix zeigt deutliche Abstufungen. Die Tubuli sind möglicherweise aus einer spiralförmigen Molekülkette aufgebaut. Das Verteilungsmuster der Organellen wird mit stereologischen Methoden untersucht. Die größte Volumendichte weisen die Aortae thoracicae mit 8% auf. Die Volumendichte der Organellen im Zytoplasma der Endothelzellen scheint mehr von der Entfernung der betreffenden Gefäßstrecke zum Herzen abzuhängen als von der Gefäßgröße. Es werden Verbindungen der Organellen zu zytoplasmatischen Membransystemen aufgezeigt. Auf Besonderheiten des Endothels, darunter Aggregationen von Ribosomen, wird hingewiesen.

---

Electron microscopic studies on specific organelles of endothelial cells in the frog (Rana temporaria)

Summary Endothelial cells of vertebrates contain specific organelles of unknown function. These organelles are studied by electron microscopy with different fixations. The organelles are rod-shaped with many variations, up to 2 in length and 0.1 to 0.5 in thickness. Their outer membrane, which is often discontinuous, is thicker than cytoplasmic membranes. The density of the matrix shows distinct gradations. The organelles contain tubules, possibly built up by a spiral molecular chain. The distribution of the organelles is investigated with stereological methods. Their volume density in endothelial cell cytoplasm appears to depend more on the distance from the heart than on vessel size, the thoracic aortae showing the highest organellae content of 8%. Connections between organelles and cytoplasmic membranes are demonstrated. Particularities of endothelium, among them aggregations of ribosomes, are pointed out.

Herrn Prof. Dr. med. F. Hammersen und Fräulein Dr. med. M. Lewerenz danke ich herzlich für Anregungen, Unterstützung und Kritik. Die Arbeit entstand während eines vom Deutschen Akademischen Austauschdienst dankenswerterweise gewährten Stipendiums; sie wurde unterstützt vom Schweizerischen Nationalfonds zur Förderung der wissenschaftlichen Forschung, Kredit-Nr. 3952.     

Effect of vesicle traps on traffic jam formation in fast axonal transport

The purpose of this paper is to develop a model for simulation of the formation of organelle traps in fast axonal transport. Such traps may form in the regions of microtubule polar mismatching. Depending on the orientation of microtubules pointing toward the trap region, these traps can accumulate either plus-end or minus-end oriented vesicles. The model predicts that the maximum concentrations of organelles occur at the boundaries of the trap regions; the overall concentration of organelles in the axon with traps is greatly increased compared to that in a healthy axon, which is expected to contribute to mechanical damages of the axon. The organelle traps induce hindrance to organelle transport down the axon; the total organelle flux down the axon with traps is found to be significantly reduced compared to that in a healthy axon.     

The zebrafish mutant vps18 as a model for vesicle‐traffic related hypopigmentation diseases

Hypopigmentation is a characteristic of several diseases associated with vesicle traffic defects, like the Hermansky–Pudlak, Chediak–Higashi, and Griscelli syndromes. Hypopigmentation is also a characteristic of the zebrafish mutant vps18^hi2499A, which is affected in the gene vps18, a component of the homotypic fusion and protein sorting complex that is involved in tethering during vesicular traffic. Vps18, as part of this complex, participates in the formation of early endosomes, late endosomes, and lysosomes. Here, we show that Vps18 is also involved in the formation of melanosomes. In the zebrafish mutant vps18^hi2499A the retroviral insertion located at exon 4 of vps18, leads to the formation of two abnormal splicing variants lacking the coding sequence for the clathrin repeat and the RING finger conserved domains. A deficiency of Vps18 in zebrafish larvae results in hepatomegaly and skin hypopigmentation. We also observed a drastic reduction in the number of melanosomes in the eye's retinal pigmented epithelium along with the accumulation of immature melanosomes. A significant reduction in the vps18^hi2499A larvae visual system capacity was found using the optokinetic response assay. We propose that the insertional mutant vps18^hi2499A can be used as a model for studying hypopigmentation diseases in which vesicle traffic problems exist.     

Temperature-induced modifications in size and pattern of microtubular organelles in a ciliate,Dileptus II. Formation and spatial arrangement of the microtubular skeleton of the oral parts

Summary Microtubular organelles formed during continuous exposure to high or low temperature were studied. Neither heat nor cold prevents formation of the microtubular skeleton in the oral parts of the ciliateDileptus. Elevated temperature causes the formation of short microtubular fibres, while the size of the oral structure is not diminished. This leads to failure in sculpturing of the cytostome. Heat-treatment may also alter the localization of the anchor site of fibres around the circumference of the basal bodies, and the orientation of the fibres. Cold-treatment evokes the formation of small mouthparts containing a lower number of organelles, although these are properly shaped and there are no deviations in the position or orientation of fibres. It seems that low temperature may suppress the rate of formation of microtubular organelles, while elevated temperature affects their patterning.Abbreviations MTOC microtubule organizing centre - T transverse fibres - B basal body - Cy cytostome - K kinetodesma - P postciliary fibre - C compound fibre - L lamina - F filamentous bundle - M monokinetid     

Extrusomes in ciliates: diversification, distribution, and phylogenetic implications

Exocytosis is, in all likelihood, an important communication method among microbes. Ciliates are highly differentiated and specialized micro-organisms for which versatile and/or sophisticated exocytotic organelles may represent important adaptive tools. Thus, in ciliates, we find a broad range of different extrusomes, i.e ejectable membrane-bound organelles. Structurally simple extrusomes, like mucocysts and cortical granules, are widespread in different taxa within the phylum. They play the roles in each case required for the ecological needs of the organisms. Then, we find a number of more elaborate extrusomes, whose distribution within the phylum is more limited, and in some way related to phylogenetic affinities. Herein we provide a survey of literature and our data on selected extrusomes in ciliates. Their morphology, distribution, and possible function are discussed. The possible phylogenetic implications of their diversity are considered.     

细胞器与细胞凋亡

细胞凋亡是由基因控制的有序生理过程,细胞内各组分在这一过程中相互协调,组成了精细的调控系统。除细胞核外,线粒体是近年发现与凋亡密切相关的细胞器,它经多种因子诱发可以释放细胞色素c等因子参与到凋亡途径中。进一步的研究发现,在一定条件下,内质网、溶酶体等也与凋亡活动有关。这些细胞器在细胞凋亡中的作用及其机制是目前的研究热点。     

Clinico‐molecular analysis of eleven patients with Hermansky–Pudlak type 5 syndrome,a mild form of HPS

Hermansky–Pudlak syndrome (HPS), first described in 1959, is a rare form of syndromic oculocutaneous albinism associated with bleeding diathesis and in some cases pulmonary fibrosis and granulomatous colitis. All 10 HPS types are caused by defects in vesicle trafficking of lysosome‐related organelles (LRO) proteins. The HPS5 protein associates with HPS3 and HPS6 to form the biogenesis of lysosome‐related organelles complex‐2 (BLOC‐2). Here, we report the clinical and genetic data of 11 patients with HPS‐5 analyzed in our laboratory. We report 11 new pathogenic variants. The 11 patients present with ocular features that are typical for albinism, with mild hypopigmentation, and with no other major complication, apart from a tendency to bleed. HPS‐5 therefore appears as a mild form of HPS, which is often clinically undistinguishable from mild oculocutaneous or ocular forms of albinism. Molecular analysis is therefore required to establish the diagnosis of this mild HPS form, which has consequences in terms of prognosis and of clinical management of the patients.