A differential genome-wide transcriptome analysis : impact of cellular copper on complex biological processes like aging and development

The regulation of cellular copper homeostasis is crucial in biology. Impairments lead to severe dysfunctions and are known to affect aging and development. Previously, a loss-of-function mutation in the gene encoding the copper-sensing and copper-regulated transcription factor GRISEA of the filamentous fungus Podospora anserina was reported to lead to cellular copper depletion and a pleiotropic phenotype with hypopigmentation of the mycelium and the ascospores, affected fertility and increased lifespan by approximately 60% when compared to the wild type. This phenotype is linked to a switch from a copper-dependent standard to an alternative respiration leading to both a reduced generation of reactive oxygen species (ROS) and of adenosine triphosphate (ATP). We performed a genome-wide comparative transcriptome analysis of a wild-type strain and the copper-depleted grisea mutant. We unambiguously assigned 9,700 sequences of the transcriptome in both strains to the more than 10,600 predicted and annotated open reading frames of the P. anserina genome indicating 90% coverage of the transcriptome. 4,752 of the transcripts differed significantly in abundance with 1,156 transcripts differing at least 3-fold. Selected genes were investigated by qRT-PCR analyses. Apart from this general characterization we analyzed the data with special emphasis on molecular pathways related to the grisea mutation taking advantage of the available complete genomic sequence of P. anserina. This analysis verified but also corrected conclusions from earlier data obtained by single gene analysis, identified new candidates of factors as part of the cellular copper homeostasis system including target genes of transcription factor GRISEA, and provides a rich reference source of quantitative data for further in detail investigations. Overall, the present study demonstrates the importance of systems biology approaches also in cases were mutations in single genes are analyzed to explain the underlying mechanisms controlling complex biological processes like aging and development

Dystonia and Paroxysmal Dyskinesias: Under-Recognized Movement Disorders in Domestic Animals? A Comparison with Human Dystonia/Paroxysmal Dyskinesias

Dystonia is defined as a neurological syndrome characterized by involuntary sustained or intermittent muscle contractions causing twisting, often repetitive movements, and postures. Paroxysmal dyskinesias are episodic movement disorders encompassing dystonia, chorea, athetosis, and ballism in conscious individuals. Several decades of research have enhanced the understanding of the etiology of human dystonia and dyskinesias that are associated with dystonia, but the pathophysiology remains largely unknown. The spontaneous occurrence of hereditary dystonia and paroxysmal dyskinesia is well documented in rodents used as animal models in basic dystonia research. Several hyperkinetic movement disorders, described in dogs, horses and cattle, show similarities to these human movement disorders. Although dystonia is regarded as the third most common movement disorder in humans, it is often misdiagnosed because of the heterogeneity of etiology and clinical presentation. Since these conditions are poorly known in veterinary practice, their prevalence may be underestimated in veterinary medicine. In order to attract attention to these movement disorders, i.e., dystonia and paroxysmal dyskinesias associated with dystonia, and to enhance interest in translational research, this review gives a brief overview of the current literature regarding dystonia/paroxysmal dyskinesia in humans and summarizes similar hereditary movement disorders reported in domestic animals

Klinische Prüfung von Homöopathika (Nosoden) in der Kontrolle von Mastitiden des Rindes (NoKoM)

In einem Milchviehbetrieb (250 Kühe) wurde eine homöopathische Trockenstellprophylaxe unter vollständigem Verzicht von antibiotischen Trockenstellern geprüft. Eine bestandsspezifische Mischnosode D30 wurde im Vergleich zu einem Placebo eingesetzt. Die Tiere wurden in wöchentlichem Abstand viermal vor dem Trockenstellen (TS), zur Abkalbung und eine Woche post partum (p.p.) mit je 5 ml des Studienpräparates peroral behandelt. Der Eutergesundheitszustand wurde zu den Behandlungen, in der 6. Woche und am Ende des 2. und 3. Laktationsmonats durch zytobakteriologische Untersuchungen erfasst. Insgesamt kamen 79 Tiere mit 314 Vierteln in die Auswertung (Verum: n=41/164; Placebo: n=58/150). In beiden Versuchsgruppen traten vergleichbare Neuinfektions-, Heilungs- und Erkrankungsraten auf. Die Neuinfektionsrate p.p. lag in der Verumgruppe bei ca. 20 % und über den Beobachtungszeitraum 5%-20% niedriger als in der Placebogruppe (p>0,05). Zum TS waren 33 Vierteln bakteriologisch positiv. Die Heilungsraten der betroffenen Viertel waren gering, und in den Behandlungsgruppen vergleichbar (p>0,05). Der vollständige Verzicht auf antibiotische Trockensteller führte nicht zur Verschlechterung der Eutergesundheit, sondern u.A. zu einer Zunahme gesunder Euterviertel. Die mittlere Tankzellzahl konnten leicht gesenkt werden. Obwohl während der Studie euterpathogene Erregern auftraten, konnte kein Anstieg an Mastitiserkrankungen festgestellt werden. Die Ergebnisse zeigen, dass der Einsatz von antibiotischen Trockenstellern deutlich reduziert werden kann, wenngleich auf ihn, vor allem in Problembetrieben, nicht ganz verzichtet werden kann. Voraussetzung bleibt aber die Optimierung des Umfeldes der Tiere. Das Konzept kann in Verbindung mit dem selektiven Einsatz von Antibiotika zusammen mit einer tierärztlichen Bestandsbetreuung auch in anderen Betrieben umgesetzt werden und liefert somit neue Erkenntnisse für Sicherung der Tiergesundheit im ökologischen Landbau

Expression of the Inhibitory CD200 Receptor Is Associated with Alternative Macrophage Activation

Classical macrophage activation is inhibited by the CD200 receptor (CD200R). Here, we show that CD200R expression was specifically induced on human in vitro polarized macrophages of the alternatively activated M2a subtype, generated by incubation with IL-4 or IL-13. In mice, peritoneal M2 macrophages, elicited during infection with the parasites Taenia crassiceps or Tryponosoma brucei brucei, expressed increased CD200R levels compared to those derived from uninfected mice. However, in vitro stimulation of mouse peritoneal macrophages and T crassiceps infection in IL-4-/- and IL-4R-/- mice showed that, in contrast to humans, induction of CD200R in mice was not IL-4 or IL-13 dependent. Our data identify CD200R as a suitable marker for alternatively activated macrophages in humans and corroborate observations of distinct species- and/or site-specific mechanisms regulating macrophage polarization in mouse and man. Copyright (C) 2009 S. Karger AG, Base

Nosoden zum Trockenstellen – eine placebokontrollierte Blindstudie

In an organic dairy herd (250 cows) a homeopathic dry cow treatment should be evaluated while an antibiotic dry cow therapy (DCT) was totally abandoned. A ran-domized placebo-controlled double blind study with a herd specific nosode (D30) was conducted. Either 5 ml of the nosode (VG) or a placebo (KG) was orally administered in weekly intervals four times before drying off, at the day of calving and seven days post partum (p.p.). The efficacy of treatment was measured according to quarter foremilk samples at the days of treatment, six weeks p.p. and at the end of the 2nd and 3rd month of lactation. Data of 129 cows with 512 quarters (VG:n=65/260; KG:n=64/252) was evaluable. New intramammary infections (IMIn) cure rates and cases of clinical mastitis in both treatment groups were nearly identical. In the VG 20% of the quarters came along with IMIn. In the KG IMIn were about 5% higher along the observation period (p>0.05). The cure rates of infected quarters were about 40% in both treatment groups (p>0.05). The complete abdication of antibiotics in DCT did not cause an impairment of udder health. Moreover the number of healthy quarters in-creased and the mean bulk milk somatic cell count decreased slightly during the study. The results of the study show that the use of antibiotics can be highly de-creased though a minimal use is indispensable, especially in herds suffering from udder health problems. Still the most essential prophylactic task is to optimize the housing conditions in the dry period and around calving. The presented dry cow man-agement in conjunction with a selective use of antibiotics can be implemented in veterinary herd health programs on other dairy farms

CD97 neutralisation increases resistance to collagen-induced arthritis in mice

Synovial tissue of rheumatoid arthritis (RA) patients is characterised by an influx and retention of CD97-positive inflammatory cells. The ligands of CD97, CD55, chondroitin sulfate B, and α5β1 (very late antigen [VLA]-5) are expressed abundantly in the synovial tissue predominantly on fibroblast-like synoviocytes, endothelium, and extracellular matrix. Based upon this expression pattern, we hypothesise CD97 expression to result in accumulation of inflammatory cells in the synovial tissue of RA patients. To determine the therapeutic effect of blocking CD97 in an animal model of RA, collagen-induced arthritis was induced in a total of 124 DBA/J1 mice. Treatment was started on day 21 (early disease) or on day 35 (longstanding disease) with the blocking hamster anti-mouse CD97 monoclonal antibody (mAb) 1B2, control hamster immunoglobulin, or NaCl, applied intraperitoneally three times a week. The paws were evaluated for clinical signs of arthritis and, in addition, examined by radiological and histological analysis. Mice receiving 0.5 mg CD97 mAb starting from day 21 had significantly less arthritis activity and hind paw swelling. Furthermore, joint damage and inflammation were reduced and granulocyte infiltration was decreased. When treatment was started on day 35, CD97 mAb treatment had similar effects, albeit less pronounced. The results support the notion that CD97 contributes to synovial inflammation and joint destruction in arthritis

Identification of neuropathology-based subgroups in multiple sclerosis using a data-driven approach

Multiple sclerosis (MS) is a heterogeneous disorder with regards to clinical presentation and pathophysiology. Stratification into biologically distinct subgroups could enhance prognostication and efficacious allocation to disease-modifying therapies. In this study, we identified MS subgroups by performing a clustering analysis on neuropathology data collected for MS donors in the Netherlands Brain Bank (NBB) autopsy cohort. The input dataset contained detailed information on white matter lesion load, the proportion of active, mixed active/inactive, inactive and remyelinating lesions, microglia morphology in these lesions, and the presence of microglial nodules, perivascular cuffs and cortical lesions for 228 donors. A factor analysis was performed to reduce noise and redundancy prior to hierarchical clustering with K-means consolidation. Four subgroups with distinct patterns of white matter lesions were identified. These were subsequently validated with additional clinical, neuropathological and genetic data. The subgroups differed with regards to disease progression and duration, the timing of motor, sensory and other relevant signs and symptoms, patterns of cortical lesions and the presence of B cells. Age at MS onset and sex, previously associated with milder forms of MS, did not differ between the subgroups; the subgroups could also not be distinguished based on the manifestation of clinical signs and symptoms. The available genetic data was used to calculate MS polygenic risk scores (PRSs) for donors included in the NBB cohort. The MS PRS did not differ between the subgroups, but was significantly correlated with the first and second dimension of the factor analysis, the latter lending genetic support to our subdivision. Taken together, these findings suggest a complex relationship between neuropathological subgroups and clinical characteristics, indicating that post-mortem cohort studies are critical to better stratify patients and understand underlying neuropathophysiological mechanisms, in order to ultimately achieve personalised medicine in MS

Leaching Hazardous Substances out of Photovoltaic Modules

Photovoltaic modules contain hazardous substances such as lead and cadmium. Under normal operation conditions, these materials will not be released into the environment. This study identifies conditions resulting in release. Our worst case study uses milled module pieces of 0.2 mm size. Depending on the pH value of water based solutions, more or less amounts of hazardous substances are leached out. Solutions with low pH values (acidic solutions) yield substantial leaching. Three different solutions simulate different environmental conditions: i) “low mineralized water” conditions, via water containing sodium hydroxide, ii) “sea water” conditions, via water containing sodium hydroxide and sodium chloride, and iii) “rainwater” conditions, via water containing acetic acid. In “rain water”-like solutions with low pH, already after a few days, around 30 % of the cadmium is leached out from milled cadmium telluride module pieces, increasing to 50 % after 56 days! In the same time, more than 15 % of lead is leached out from c-Si module pieces. Tellurium elutes in the range of 30 to 40 % with a weak dependence on the pH value of the solution indicating an instability of the compound cadmiumtelluride out of the cadmiumtelluride modules. Most of the extractions increase during several weeks of measurement. Therefore, the usual one-day-elution test does not give enough information. Meaningful leaching experiments should last for at least ten days