Isolation, establishment, and characterization of ex vivo equine melanoma cell cultures

Gray horses spontaneously develop metastatic melanomas that resemble human disease, and this is often accompanied with metastasis to other organs. Unlike in other species, the establishment of primary equine melanoma cultures that could be used to develop new therapeutic approaches has remained a major challenge. The purpose of the study was to develop a protocol for routine isolation and cultivation of primary equine melanocytes. Melanoma tissues were excised from 13 horses under local anesthesia, mainly from the perianal area. The melanoma cells were isolated from the melanoma tissue by serial enzymatic digestion using dispase and collagenase. Out of the 13 excised melanomas, cell cultures from eight melanomas were established, which corresponded to a success rate 62%. These cells showed different degrees of melanin pigmentation. Characterization of these cells using confocal microscopy, FACs analysis and western blotting showed that they expressed melanoma-associated antigens; Melan-A, MAGE-1, and MAGE-3, and PCNA expression was higher in fast-proliferating isolates. The protocol we developed and established proved successful for routine isolation and cultivation of primary equine melanoma cells. This method provided a large number of primary equine melanoma cells that could be used to study new therapeutic approaches for treatment of equine melanoma

Zuordnungen in Bewegung : Geschlecht und sexuelle Orientierung quer durch die Disziplinen

Durch aktuelle gesellschaftspolitische und rechtliche Debatten um geschlechtliche und sexuelle Vielfalt ist einiges in Bewegung geraten: Tradierte (Zu-)Ordnungen von Geschlecht sind durchlässiger geworden, strikte Regulierungen von Sexualität(en) sind aufgebrochen. Soziale Wirklichkeiten scheinen nun pluralisiert und neue Chancen für selbstbestimmte Lebens­weisen eröffnet. Zugleich haben institutionalisierte Vorstellungen von binären Ge­schlech­terdifferenzen und von Heterosexualität Spuren hinterlassen, die als ‚Einschreibungen’ der sozialen Verhältnisse weiter existieren. Der vorliegende Band ist das Ergebnis interdisziplinärer Reflexionen in den Sozial- und Geisteswissenschaften. Die Beiträge geben einen Einblick in unterschiedliche disziplinäre Perspektiven auf Gender- und Sexualitäts(zu)ordnungen. Sie spannen die Breite der For­schungsfelder mit ihrem jeweiligen Bezug zu Alltagspraktiken auf; vor dem Hintergrund der Fachkulturen und wissenschaftlichen Entwicklungen beleuchten sie begriffliche Konzepte, Fragestellungen sowie Arbeitsweisen und reflektieren vor allem auch die Gleichzeitigkeit von Kontinuität, Wandel und neuen Normierungsprozessen.Der vorliegende Band ist das Ergebnis interdisziplinärer Reflexionen in den Sozial- und Geisteswissenschaften. Die Beiträge geben einen Einblick in unterschiedliche disziplinäre Perspektiven auf Gender- und Sexualitäts(zu)ordnungen. Sie spannen die Breite der For­schungsfelder mit ihrem jeweiligen Bezug zu Alltagspraktiken auf; vor dem Hintergrund der Fachkulturen und wissenschaftlichen Entwicklungen beleuchten sie begriffliche Konzepte, Fragestellungen sowie Arbeitsweisen und reflektieren vor allem auch die Gleichzeitigkeit von Kontinuität, Wandel und neuen Normierungsprozesse

Isolation, establishment, and characterization of ex vivo equine melanoma cell cultures

Gray horses spontaneously develop metastatic melanomas that resemble human disease, and this is often accompanied with metastasis to other organs. Unlike in other species, the establishment of primary equine melanoma cultures that could be used to develop new therapeutic approaches has remained a major challenge. The purpose of the study was to develop a protocol for routine isolation and cultivation of primary equine melanocytes. Melanoma tissues were excised from 13 horses under local anesthesia, mainly from the perianal area. The melanoma cells were isolated from the melanoma tissue by serial enzymatic digestion using dispase and collagenase. Out of the 13 excised melanomas, cell cultures from eight melanomas were established, which corresponded to a success rate 62%. These cells showed different degrees of melanin pigmentation. Characterization of these cells using confocal microscopy, FACs analysis and western blotting showed that they expressed melanoma-associated antigens; Melan-A, MAGE-1, and MAGE-3, and PCNA expression was higher in fast-proliferating isolates. The protocol we developed and established proved successful for routine isolation and cultivation of primary equine melanoma cells. This method provided a large number of primary equine melanoma cells that could be used to study new therapeutic approaches for treatment of equine melanomas

Climate response to projected changes in short-lived species under an A1B scenario from 2000-2050 in the GISS climate model

We investigate the climate forcing from and response to projected changes in short-lived species and methane under the A1B scenario from 2000-2050 in the GISS climate model. We present a meta-analysis of new simulations of the full evolution of gas and aerosol species and other existing experiments with variations of the same model. The comparison highlights the importance of several physical processes in determining radiative forcing, especially the effect of climate change on stratosphere-troposphere exchange, heterogeneous sulfate-nitrate-dust chemistry, and changes in methane oxidation and natural emissions. However, the impact of these fairly uncertain physical effects is substantially less than the difference between alternative emission scenarios for all short-lived species. The net global mean annual average direct radiative forcing from the short-lived species is .02 W/m{sup 2} or less in our projections, as substantial positive ozone forcing is largely offset by negative aerosol direct forcing. Since aerosol reductions also lead to a reduced indirect effect, the global mean surface temperature warms by {approx}0.07 C by 2030 and {approx}0.13 C by 2050, adding 19% and 17%, respectively, to the warming induced by long-lived greenhouse gases. Regional direct forcings are large, up to 3.8 W/m{sup 2}. The ensemble-mean climate response shows little regional correlation with the spatial pattern of the forcing, however, suggesting that oceanic and atmospheric mixing generally overwhelms the effect of even large localized forcings. Exceptions are the polar regions, where ozone and aerosols may induce substantial seasonal climate changes

The Mouse Cytomegalovirus Gene m42 Targets Surface Expression of the Protein Tyrosine Phosphatase CD45 in Infected Macrophages

The receptor-like protein tyrosine phosphatase CD45 is expressed on the surface of cells of hematopoietic origin and has a pivotal role for the function of these cells in the immune response. Here we report that following infection of macrophages with mouse cytomegalovirus (MCMV) the cell surface expression of CD45 is drastically diminished. Screening of a set of MCMV deletion mutants allowed us to identify the viral gene m42 of being responsible for CD45 down-modulation. Moreover, expression of m42 independent of viral infection upon retroviral transduction of the RAW264.7 macrophage cell line led to comparable regulation of CD45 expression. In immunocompetent mice infected with an m42 deletion mutant lower viral titers were observed in all tissues examined when compared to wildtype MCMV, indicating an important role of m42 for viral replication in vivo. The m42 gene product was identified as an 18 kDa protein expressed with early kinetics and is predicted to be a tailanchored membrane protein. Tracking of surface-resident CD45 molecules revealed that m42 induces internalization and degradation of CD45. The observation that the amounts of the E3 ubiquitin ligases Itch and Nedd4 were diminished in cells expressing m42 and that disruption of a PY motif in the N-terminal part of m42 resulted in loss of function, suggest that m42 acts as an activator or adaptor for these Nedd4-like ubiquitin ligases, which mark CD45 for lysosomal degradation. In conclusion, the down-modulation of CD45 expression in MCMV-infected myeloid cells represents a novel pathway of virus-host interaction

Battling demons with medical authority: werewolves, physicians and rationalization

Werewolves and physicians experienced their closest contact in the context of early modern witch and werewolf trials. For medical critics of the trials, melancholic diseases served as reference points for medical explanations of both individual cases and werewolf beliefs in general. This paper attempts to construct a conceptual history of werewolf beliefs and their respective medical responses. After differentiating the relevant terms, pre-modern werewolf concepts and medical lycanthropy are introduced. The early modern controversy between medical and demonological explanations forms the main part of this study. The history of werewolves and their medical explanations is then traced through to present times. An important point of discussion is to what extent the physicians’ engagements with werewolves can be characterized as rationalization

Kynanthropy: canine madness in Byzantine late antiquity

Those afflicted bark like dogs, scramble on all fours and loiter around graveyards – canine madness, referred to as kynanthropy, was an illness concept in its own right in the medicine of late antiquity. At roughly the same time as the medical description produced by Aëtius of Amida, the Syrian chronicler John of Ephesus, also from Amida, reported an epidemic of dog-like madness sweeping his home town in ad 560. The symptoms are identical and both authors are from Amida – what is the connection between the two depictions? In addition to the history of the medical concept, the example of the canine madness of Amida and its cultural embedding allows us to contextualize and interpret the significance of dog-like behaviour for the people of the sixth century ad

Poisoning, Ergotism, Mass Psychosis: Writing a History of Ancient Epidemics Beyond Infectious Diseases

For the last 100 years, the modern concept of epidemics as contagious diseases caused by pathogenic agents or microorganisms entering the body has not only dominated present thinking about epidemics but highly influenced historiographical study of past disease as well. In the case of Greek and Roman antiquity, this led to extensive and thorough scholarly work on epidemics fitting the pattern of infectious diseases while incompatible cases were put aside notwithstanding that by ancient definition they were epidemics of the same quality: illness that affects many individuals of the same community at the same time. This includes cases retrospectively explained as mass poisoning, ergotism, and mass hysteria. This article discusses the methodological problem of disparate definitions of modern and ancient epidemics and argues for broadening the source base in the study of ancient epidemics to include accounts of diseases that do not fit into the modern mould of infectious disease. To demonstrate the benefit of this suggestion, two disregarded later ancient epidemics drawn from relatively unknown patristic sources are introduced, which have been explained as fungal poisoning, ergotism, or mass psychosis in the past