Characteristics of Exosomes and the Vascular Landscape Regulate Exosome Sequestration by Peripheral Tissues and Brain

Exosomes mediate intercellular communication, shuttling messages between cells and tissues. We explored whether exosome tissue sequestration is determined by the exosomes or the tissues using ten radiolabeled exosomes from human or murine, cancerous or noncancerous cell lines. We measured sequestration of these exosomes by the liver, kidney, spleen, and lung after intravenous injection into male CD-1 mice. Except for kidney sequestration of three exosomes, all exosomes were incorporated by all tissues, but sequestration levels varied greatly among exosomes and tissues. Species of origin (mouse vs. human) or source (cancerous vs. noncancerous cells) did not influence tissue sequestration. Sequestration of J774A.1 exosomes by liver involved the mannose-6 phosphate (M6P) receptor. Wheatgerm agglutinin (WGA) or lipopolysaccharide (LPS) treatments enhanced sequestration of exosomes by brain and lung but inhibited sequestration by liver and spleen. Response to LPS was not predictive of response to WGA. Path and heat map analyses included our published results for brain and found distinct clusters among the exosomes and the tissues. In conclusion, we found no evidence for a universal binding site controlling exosome-tissue interactions. Instead, sequestration of exosomes by tissues is differentially regulated by both exosomes and tissues and may be stimulated or inhibited by WGA and inflammation.     

Photorhabdus luminescens TccC3 Toxin Targets the Dynamic Population of F-Actin and Impairs Cell Cortex Integrity

Due to its essential role in cellular processes, actin is a common target for bacterial toxins. One such toxin, TccC3, is an effector domain of the ABC-toxin produced by entomopathogenic bacteria of Photorhabdus spp. Unlike other actin-targeting toxins, TccC3 uniquely ADP-ribosylates actin at Thr-148, resulting in the formation of actin aggregates and inhibition of phagocytosis. It has been shown that the fully modified F-actin is resistant to depolymerization by cofilin and gelsolin, but their effects on partially modified actin were not explored. We found that only F-actin unprotected by tropomyosin is the physiological TccC3 substrate. Yet, ADP-ribosylated G-actin can be produced upon cofilin-accelerated F-actin depolymerization, which was only mildly inhibited in partially modified actin. The affinity of TccC3-ADP-ribosylated G-actin for profilin and thymosin-β4 was weakened moderately but sufficiently to potentiate spontaneous polymerization in their presence. Interestingly, the Arp2/3-mediated nucleation was also potentiated by T148-ADP-ribosylation. Notably, even partially modified actin showed reduced bundling by plastins and α-actinin. In agreement with the role of these and other tandem calponin-homology domain actin organizers in the assembly of the cortical actin network, TccC3 induced intense membrane blebbing in cultured cells. Overall, our data suggest that TccC3 imposes a complex action on the cytoskeleton by affecting F-actin nucleation, recycling, and interaction with actin-binding proteins involved in the integration of actin filaments with each other and cellular elements.     

Lagerung von Kartoffeln in kontrollierter Atmosphäre

Storage of potatoes in controlled atmosphere. The interim results obtained from trials with CA-storage are reported. Storage atmospheres with low O₂ and increased CO₂ contents largely inhibit the sprout growth of potatoes kept at a temperature of 9 °C so that chemical sprout suppressants may possibly be dispensable. Storage atmospheres with too high a CO₂ content cause an intense rise of the saccharose content and lead to increased decay. With tolerable storage atmospheres these disadvantages can be avoided. The remaining questions shall be answered in further trials which shall also help to bring CA-storage to perfection for its application in practice.     

明式家具的丹麦知音

<正>即使在北欧人力费用极高的情况下,卡尔·汉森·桑仍以近乎苛刻的标准进行手工制作,并将这份永恒的经典和淡然的精致传递给全世界的使用者。丹麦设计界流传着这样一则故事:1807年,英国轰炸丹麦首都哥本哈根。时任丹麦国王下令从斯德哥尔摩南部到汉堡北部,用一切办法遍植橡树以躲避战争的创伤,并规定:每棵树都不可被砍掉,除非新种一棵。结果,战争的炮火并未殃及丹麦,这些橡树幸运存活了下来,年复一年,长成茂密森林。闻着木香,我们来到哥本哈根繁华都市的心脏处——自1780年起就开始引领丹麦设计与工艺时尚的Bredgade大街,百年品牌卡尔·汉森·桑(Carl Hansen&Son)全球旗舰店就坐落于此。     

Isobaric thermal expansivities of mixtures ofm-cresol and quinoline from 0.1 to 400 MPa at 303 to 503 K

Isobaric thermal expansivities, α _p (p, T), of five binary mixtures ofm-cresol with quinoline (0.1499, 0.2998, 0.5005, 0.6325, and 0.8501 mol fraction ofm-cresol) were measured in a pressure-controlled scanning calorimeter over the pressure range from just above the saturation vapor pressures to 400 MPa, and at 303.15, 353.15, 403.15, 453.15, and 503.15 K. Molecular association ofm-cresol with itself and ofm-cresol with quinoline exerts large effects on the pressure and temperature behavior of α _p isotherms. The extent of association changes significantly with conditions in all except the 2∶1 mixture as demonstrated by the crossing of isotherms at lower pressures as the temperature increases. In the 2∶1m-cresol quinoline mixture the extent of association is not perturbed significantly by temperature change and the mixture behaves like a simple liquid, exhibiting a unique crossing point of α _p isotherms.     

Production and Characterization of Peptide Antibodies to the C-Terminal of Frameshifted Calreticulin Associated with Myeloproliferative Diseases

Myeloproliferative Neoplasms (MPNs) constitute a group of rare blood cancers that are characterized by mutations in bone marrow stem cells leading to the overproduction of erythrocytes, leukocytes, and thrombocytes. Mutations in calreticulin (CRT) genes may initiate MPNs, causing a novel variable polybasic stretch terminating in a common C-terminal sequence in the frameshifted CRT (CRTfs) proteins. Peptide antibodies to the mutated C-terminal are important reagents for research in the molecular mechanisms of MPNs and for the development of new diagnostic assays and therapies. In this study, eight peptide antibodies targeting the C-terminal of CRTfs were produced and characterised by modified enzyme-linked immunosorbent assays using resin-bound peptides. The antibodies reacted to two epitopes: CREACLQGWTE for SSI-HYB 385-01, 385-02, 385-03, 385-04, 385-07, 385-08, and 385-09 and CLQGWT for SSI-HYB 385-06. For the majority of antibodies, the residues Cys¹, Trp⁹, and Glu¹¹ were essential for reactivity. SSI-HYB 385-06, with the highest affinity, recognised recombinant CRTfs produced in yeast and the MARIMO cell line expressing CRTfs when examined in Western immunoblotting. Moreover, SSI-HYB 385-06 occasionally reacted to CRTfs from MPN patients when analysed by flow cytometry. The characterized antibodies may be used to understand the role of CRTfs in the pathogenesis of MPNs and to design and develop new diagnostic assays and therapeutic targets.     

Structural characterization of the rhenium(v) oxo complex of mercaptoacetyltriglyclne in its dianionic form

Dianionic [MO(MAG(3))](2-)(MAG(3) = penta-anionic form of mercaptoacetyltriglycine, M = (186)Re, (99m)Tc) complexes have important applications in nuclear medicine. In vivo the complexes have a deprotonated carboxyl group that is important to their biodistribution. The solid-state structures of (99)Tc and Re complexes with mercaptoacetyltriglycine reported previously are monoanions with protonated carboxyl groups. In the present work, we report the preparation and X-ray crystal structure of Na(2)[ReO(MAG(3))].5H(2)O (1), which contains the physiologically relevant dianion. The dianion is a distorted square pyramid with the nitrogen and sulphur donor atoms forming the base and the oxo ligand at the apex. The terminal carboxyl group is deprotonated, uncoordinated and has a syn orientation with respect to the oxo ligand. The syn conformation of the dianion in 1 differs in conformation from the anti-monoanion in [Bu(4)N][ReO(MAG(3)H)] but is similar to the syn-monoanion in [Ph(4)P][(99)TcO(MAG(3) H)].