Mutation of Threonine 34 in Mouse Podoplanin-Fc Reduces CLEC-2 Binding and Toxicity in Vivo While Retaining Anti-lymphangiogenic Activity

The lymphatic system plays an important role in cancer metastasis and inhibition of lymphangiogenesis could be valuable in fighting cancer dissemination. Podoplanin (Pdpn) is a small, transmembrane glycoprotein expressed on the surface of lymphatic endothelial cells (LEC). During mouse development, binding of Pdpn to the C-type lectin-like receptor 2 (CLEC-2) on platelets is critical for the separation of the lymphatic and blood vascular systems. Competitive inhibition of Pdpn functions with a soluble form of the protein, Pdpn-Fc, leads to reduced lymphangiogenesis in vitro and in vivo. However, the transgenic overexpression of human Pdpn-Fc in mouse skin causes disseminated intravascular coagulation due to platelet activation via CLEC-2. In the present study, we produced and characterized a mutant form of mouse Pdpn-Fc, in which threonine 34, which is considered essential for CLEC-2 binding, was mutated to alanine (PdpnT34A-Fc). Indeed, PdpnT34A-Fc displayed a 30-fold reduced binding affinity for CLEC-2 compared with Pdpn-Fc. This also translated into fewer side effects due to platelet activation in vivo. Mice showed less prolonged bleeding time and fewer embolized vessels in the liver, when PdpnT34A-Fc was injected intravenously. However, PdpnT34A-Fc was still as active as wild-type Pdpn-Fc in inhibiting lymphangiogenesis in vitro and also inhibited lymphangiogenesis in vivo. These data suggest that the function of Pdpn in lymphangiogenesis does not depend on threonine 34 in the CLEC-2 binding domain and that PdpnT34A-Fc might be an improved inhibitor of lymphangiogenesis with fewer toxic side effects.     

Can We Use Tree Rings of Black Alder to Reconstruct Lake Levels? A Case Study for the Mecklenburg Lake District,Northeastern Germany

In this study, we explore the potential to reconstruct lake-level (and groundwater) fluctuations from tree-ring chronologies of black alder (Alnus glutinosa L.) for three study lakes in the Mecklenburg Lake District, northeastern Germany. As gauging records for lakes in this region are generally short, long-term reconstructions of lake-level fluctuations could provide valuable information on past hydrological conditions, which, in turn, are useful to assess dynamics of climate and landscape evolution. We selected black alder as our study species as alder typically thrives as riparian vegetation along lakeshores. For the study lakes, we tested whether a regional signal in lake-level fluctuations and in the growth of alder exists that could be used for long-term regional hydrological reconstructions, but found that local (i.e. site-specific) signals in lake level and tree-ring chronologies prevailed. Hence, we built lake/groundwater-level reconstruction models for the three study lakes individually. Two sets of models were considered based on (1) local tree-ring series of black alder, and (2) site-specific Standardized Precipitation Evapotranspiration Indices (SPEI). Although the SPEI-based models performed statistically well, we critically reflect on the reliability of these reconstructions, as SPEI cannot account for human influence. Tree-ring based reconstruction models, on the other hand, performed poor. Combined, our results suggest that, for our study area, long-term regional reconstructions of lake-level fluctuations that consider both recent and ancient (e.g., archaeological) wood of black alder seem extremely challenging, if not impossible.     

Retroviral-mediated gene transfer as an effective tool for the in vitro genetic transformation of chicken embryonic cells and production of transgenic chickens

The data on the in vitro and in vivo (into embryonic disk) retroviral-mediated transfer of genetic information into chicken embryonic cells are presented. The estimated transformation frequency of the cultured target cells constituted 8 × 10^?4 to 5 × 10^?3. A transgenic rooster, carrying recombinant DNA in blood, heart, liver, and intestine cells, was obtained.     

Vorkommen und Funktion sensibler Erregungssubstanzen und sie abbauender Fermente im Tierreich

Zusammenfassung Im Zentralnervensystem der Wirbeltiere wird die Erregung sensibler Nerven durch eine eigene, von ihnen gebildete Erregungssubstanz vermittelt, welche durch ein Ferment rasch wieder abgebaut wird (Hellauer und Umrath ⁴⁹). Das Ferment läßt sich durch Strychnin, Pikrotoxin, Brucin und Cardiazol hemmen (Hellauer und Umrath ⁵⁰). Diese Pharmaka bewirken daher Erregung und charakteristische Krämpfe.Durch einen neuen Test an Bienen konnte gezeigt werden, daß die sensiblen Nerven der Arthropoden eine von der der Wirbeltiere etwas verschiedene Erregungssubstanz bilden. Ihr fermentativer Abbau wird durch Pikrotoxin und Cardiazol gehemmt, nicht aber durch Strychnin.An einer großen Anzahl von Arten aus dem ganzen Tierreich wurden die Reaktionen auf Strychnin, Pikrotoxin und Cardiazol geprüft. Es zeigte sich, daß bestimmte Tiergruppen jeweils verschiedene sensible Erregungssubstanzen und dementsprechend verschiedene abbauende Fermente besitzen. Es ergaben sich interessante Beziehungen zur Systematik: Die Deuterostomier (Vertebraten, Tunikaten, Echinodermen, Chätognathen) erwiesen sich hinsichtlich der sensiblen Erregungssubstanz und des sie abbauenden Fermentes als einheitliche Gruppe (Hemmung des Abbaues durch Strychnin, Pikrotoxin und Cardiazol). Einheitlich sind auch die Arthropoden (Hemmung durch Pikrotoxin und Cardiazol). Bei Mollusken hemmt ausschließlich Strychnin, das auch bei verschiedenen anderen Gruppen der Protostomier diese Wirkung hat, wenn auch zum Teil schwächer. Pikrotoxin hemmt außer bei Deuterostomiern und Arthropoden nur noch bei Turbellarien und Nemertinen, bei denen auch Strychnin und Cardiazol wirksam sind.Die sensible Erregungssubstanz der Clitellaten ist Acetylcholin.Bei Cölenteraten erwiesen sich die Pharmaka als unwirksam. Bei Ciliaten ist das Vorkommen einer sensiblen Erregungssubstanz mit fermentativem Abbau (Hemmung durch Strychnin) möglich.Die Wirkungsweise von Strychnin, Pikrotoxin, Brucin und Cardiazol wird besprochen.     

Über die Lagerung der Chromoplasten (Beobachtungen an pflanzlichen Haaren)

Zusammenfassung Im Gegensatz zu den Chloroplasten können die Chromoplasten (Untersuchungen an den Haaren vonCucurbita undGaillardia) nicht zu phototaktischen Orientierungsbewegungen veranlaßt werden. Wie die Chloroplasten oder andere zur Assimilation befähigte Plastiden lassen sich indessen die Chromoplasten vonGaillardia durch Behandlung mit wasserentziehenden Mitteln zu Bewegungen veranlassen: sie suchen Gürtelstellung an den Längswänden der Haare auf. Die Häufung der Chromoplasten am Zellkern und ihre Abwanderung von den wandständigen Plasmabelägen zu den Plasmasträngen sowie ihre Häufung an den Querwänden, insbesondere an ihren Mittelfeldern lassen sich vorläufig nicht durch Außenweltsfaktoren experimentell hervorrufen. — Die Chromoplasten vonCucurbita ließen sich durch Behandlung mit wasserentziehenden Mitteln nicht zur Verlagerung bringen.     

New Aradidae from Ecuador (Hemiptera,Heteroptera, Aradidae)

As an addition to the presently poorly known aradid fauna of Ecuador, 3 new genera and 4 new species are described: Osellaptera setifera gen. n., sp. n.; Kormilevia ecuadoriana sp. n. both belonging to Mezirinae; and Carventinae Cotopaxicoris cruciatus gen. n., sp. n. and Onorecoris piceus gen. n., sp. n. An updated key is provided for all species of the Neotropical genus Kormilevia Usinger & Matsuda, 1959.     

Depicting the Spatial Distribution of Proteins in Human Tumor Tissue Combining SELDI and MALDI Imaging and Immunohistochemistry

Carcinoma tissue consists of not only tumor cells but also fibroblasts, endothelial cells or vascular structures, and inflammatory cells forming the supportive tumor stroma. Therefore, the spatial distribution of proteins that promote growth and proliferation in these complex functional units is of high interest. Matrix-assisted laser desorption/ionization imaging mass spectrometry is a newly developed technique that generates spatially resolved profiles of protein signals directly from thin tissue sections. Surface-enhanced laser desorption/ionization mass spectrometry (MS)combined with tissue microdissection allows analysis of defined parts of the tissue with a higher sensitivity and a broader mass range. Nevertheless, both MS-based techniques have a limited spatial resolution. IHC is a technique that allows a resolution down to the subcellular level. However, the detection and measurement of a specific protein expression level is possible only by semiquantitative methods. Moreover, prior knowledge about the identity of the proteins of interest is necessary. In this study, we combined all three techniques to gain highest spatial resolution, sensitivity, and quantitative information. We used frozen tissue from head and neck tumors and chose two exemplary proteins (HNP1–3 and S100A8) to highlight the advantages and disadvantages of each technique. It could be shown that the combination of these three techniques results in congruent but also synergetic data. (J Histochem Cytochem 58:929–937, 2010)     

Influence of chronic antigen exposure on the metabolism of PGH2 by microsomal preparations of airway and parenchymal tissues in the sheep

The effects of repeated antigen exposure on the synthesis of mediators by lung tissues are not well understood. To investigate the influence of antigen challenge on the synthesis of prostaglandins by central airway and peripheral lung tissues, fourteen sensitive sheep underwent biweekly exposure to aerosolized $\text{Ascaris suu}$ antigen (7) or saline (7). Following the fifth exposure, microsomal and high speed supernatant fractions were prepared from trachealis muscle and lung parenchyma. Synthesis of thromboxane (TX) A₂, prostaglandin (PG) D₂ and PGI₂ from the PG endoperoxide intermediate, PGH₂, was assayed over a range of substrate concentrations from 3–200 uM. Synthesis of PGI₂ by trachealis microsomes was approximately 5-fold greater than that of TXA₂. PGI₂ and TXA₂ production was identical in tracheal preparations from Ascaris- and saline-exposed animals. In parenchymal tissues, where TXA₂ production predominated over PGI₂ by 9-fold, preparations from Ascaris- exposed animals synthesized 50% more TXA₂ than controls at PGH₂ concentrations of 25 uM and above, whereas synthesis of PGI₂ and PGD₂ were similar in preparations from both groups of animals. The density of pulmonary mast cells was decreased by 21% in the Ascaris group, whereas polymorphonuclear leukocyte density was unchanged. These results demonstrate the differential synthesis of TXA₂ and PGI₂ in central airways and peripheral lung regions of the sheep. They further indicate that repeated exposure of the airways to antigen selectively enhances TXA₂ synthesis in the lung periphery of sensitized animals. The site of this increased enzymatic activity, whether in resident cells or newly-infiltrated cells, has not been determined.