Influence of Atrial Natriuretic Peptide on inflammatory pathways in the lung

The cardiovascular hormone ANP is known to exert anti-inflammatory properties in macrophages and endothelial cells. This work provides new insight into the inflammatory signalling pathways influenced by the ANP in the lung. For these purposes, the effects of ANP on both alveolar epithelial cells and a model of LPS-induced lung inflammation were characterized. In alveolar epithelial cells, ANP was shown to inhibit the activation of two major transcription factors, NF-kB and AP-1, in response to TNFa. Astonishingly, this did not result in a reduced expression of the adhesion molecule ICAM-1. ANP was also capable to diminish the activation of AP-1 and NF-kB in lung tissue in vivo using a mouse model of LPS-induced septic shock. The inhibition of NF-kB activation was caused by a delayed phosphorylation and subsequent degradation of IkBa. In addition, ANP treatment elevated total protein levels of IkBa. p38 MAPK and Akt are important mediators in LPS-induced signalling. We demonstrated an activation of these kinases in lung tissue in response to i.p. LPS challenge. ANP treatment was able to lessen this activation. Furthermore, exclusive ANP treatment resulted in an increased p38 MAPK activation, which might contribute to the observed impact on other pathways. ICAM-1 expression was not impaired in whole lung tissue. ANP strongly decreased TNFa serum levels dose-dependently, but had only a slight effect on TNFa tissue levels. Interestingly, TNFa mRNA expression was not significantly reduced. Taken together this work demonstrates that ANP is able to diminish several important inflammatory pathways which are involved in the development of acute respiratory distress syndrome in LPS-induced sepsis

Influence of Atrial Natriuretic Peptide on inflammatory pathways in the lung

The cardiovascular hormone ANP is known to exert anti-inflammatory properties in macrophages and endothelial cells. This work provides new insight into the inflammatory signalling pathways influenced by the ANP in the lung. For these purposes, the effects of ANP on both alveolar epithelial cells and a model of LPS-induced lung inflammation were characterized. In alveolar epithelial cells, ANP was shown to inhibit the activation of two major transcription factors, NF-kB and AP-1, in response to TNFa. Astonishingly, this did not result in a reduced expression of the adhesion molecule ICAM-1. ANP was also capable to diminish the activation of AP-1 and NF-kB in lung tissue in vivo using a mouse model of LPS-induced septic shock. The inhibition of NF-kB activation was caused by a delayed phosphorylation and subsequent degradation of IkBa. In addition, ANP treatment elevated total protein levels of IkBa. p38 MAPK and Akt are important mediators in LPS-induced signalling. We demonstrated an activation of these kinases in lung tissue in response to i.p. LPS challenge. ANP treatment was able to lessen this activation. Furthermore, exclusive ANP treatment resulted in an increased p38 MAPK activation, which might contribute to the observed impact on other pathways. ICAM-1 expression was not impaired in whole lung tissue. ANP strongly decreased TNFa serum levels dose-dependently, but had only a slight effect on TNFa tissue levels. Interestingly, TNFa mRNA expression was not significantly reduced. Taken together this work demonstrates that ANP is able to diminish several important inflammatory pathways which are involved in the development of acute respiratory distress syndrome in LPS-induced sepsis

Sprachbewusst unterrichten. Handreichung für den Unterricht an Grund-, Haupt-, Ober- und Realschulen

Sprachliche Bildung in allen Fächern gehört zu den Kernaufgaben der Grund- und weiterführenden Schulen. Unterschiede in den sprachlichen Kompetenzen der Schülerinnen und Schüler erfordern einen Unterricht, der sich neben der fachlichen Vermittlung zugleich sprachsensibel verhält. Dies wiederum verlangt von den Lehrpersonen entsprechende Konzepte, mit denen sie den sprachlichen Herausforderungen ihrer Schülerinnen und Schüler konstruktiv begegnen können. Die Handreichung richtet sich deshalb an Dozentinnen und Dozenten aller Fächer in der Hochschullehre und in der Lehrerfortbildung sowie an Lehrerinnen und Lehrer, die sich im Selbststudium zu Fragen der sprachlichen Bildung weiterqualifizieren möchten. Neben einem theoretischen Überblick über aktuelle Aspekte sprachsensiblen Unterrichts werden exemplarische Seminarkonzepte aus der universitären Lehrerfortbildung vorgestellt. Das Material der Handreichung ist im Rahmen des Verbundprojektes „UMBRÜCHE GESTALTEN. Sprachenförderung und -bildung als integrale Bestandteile innovativer Lehramtsausbildung in Niedersachsen“ entwickelt, erprobt und evaluiert worden

Directed PCR-free engineering of highly repetitive DNA sequences

Background: Highly repetitive nucleotide sequences are commonly found in nature e.g. in telomeres, microsatellite DNA, polyadenine (poly(A)) tails of eukaryotic messenger RNA as well as in several inherited human disorders linked to trinucleotide repeat expansions in the genome. Therefore, studying repetitive sequences is of biological, biotechnological and medical relevance. However, cloning of such repetitive DNA sequences is challenging because specific PCR-based amplification is hampered by the lack of unique primer binding sites resulting in unspecific products.Results: For the PCR-free generation of repetitive DNA sequences we used antiparallel oligonucleotides flanked byrestriction sites of Type IIS endonucleases. The arrangement of recognition sites allowed for stepwise and seamless elongation of repetitive sequences. This facilitated the assembly of repetitive DNA segments and open reading frames encoding polypeptides with periodic amino acid sequences of any desired length. By this strategy wecloned a series of polyglutamine encoding sequences as well as highly repetitive polyadenine tracts. Such repetitive sequences can be used for diverse biotechnological applications. As an example, the polyglutamine sequences were expressed as His6-SUMO fusion proteins in Escherichia coli cells to study their aggregation behavior in vitro. The His6-SUMO moiety enabled affinity purification of the polyglutamine proteins, increased their solubility, and allowed controlled induction of the aggregation process. We successfully purified the fusions proteins and provide an example for their applicability in filter retardation assays.Conclusion: Our seamless cloning strategy is PCR-free and allows the directed and efficient generation of highlyrepetitive DNA sequences of defined lengths by simple standard cloning procedures

Prisoner at home or active community participant? : exploring factors that influence public transport service provision for persons with disabilities in the eThekwini Municipality.

Thesis (M.Dev.Studies)-University of KwaZulu-Natal, Durban, 2013.This study explores the factors influencing public transport service provision for persons with disabilities in eThekwini, South Africa. It was motivated by an awareness of the many barriers that persons with disabilities face to participate fully within society. The study recognises that transport is a means to an end, and an end in itself. It is a complicated system and therefore it is explored in this study from the perspectives of the various stakeholders, namely persons with disabilities; the operators and owners of public transport systems; and the city officials, consultants and experts in the field. The results indicate that persons with disabilities are excluded from public transport and experience challenges in each step of the travel chain. Themes highlighted include the power of money, the attitude of the transport operators and passengers, as well as the inaccessibility of the vehicles and built environment. This in turn contributes to occupational marginalization, occupational deprivation, occupational apartheid and limited occupational choice, which is related to not being able to pursue various occupations, being isolated, feeling powerless and having a reduced sense of well-being. The eThekwini Municipality is in the process of planning and implementing the Integrated Rapid Public Transport Network, however there are concerns about this future system and the lack of Accelerated Modal Upgrading and other measures to improve the current system. There needs to be a greater commitment to addressing public transport service provision holistically, so that persons with disabilities can fully participate as citizens in society

Atrial Natriuretic Peptide, a Regulator of Nuclear Factor-κB Activation in Vivo

Natriuretic peptides (NPs) comprise a family of vasoactive hormones that play important roles in the regulation of cardiovascular and renal homeostasis. Along this line, atrial NP (ANP) (international non-proprietary name: carperitide, HANP) is an approved drug for the treatment of acute heart failure. In recent years, evidence has been given that the NP system possesses a far broader biological spectrum than the regulation of blood pressure and volume homeostasis. In fact, a substantial amount of in vitro work indicates that ANP affects important inflammatory processes and signaling pathways. Quite surprisingly, however, no information exists on the in vivo antiinflammatory potential and signaling of ANP. We show here that pretreatment of lipopolysaccharide (Salmonella abortus equi, 2.5 mg/kg)-challenged mice with ANP (5μg/kg iv, 15 min) rapidly inhibits nuclear factor-κB activation via inhibition of phosphorylation and degradation of the IκB-α protein. ANP also reduces Akt activation upon lipopolysaccharide injection. In ANP-pretreated mice, the increase of TNF-α serum concentration is markedly prevented; most importantly, the survival of these animals improved. These findings demonstrate both in vitro and in vivo an antiinflammatory profile of ANP that deserves to be further investigated in a therapeutic perspective

Computer-assisted surgery for placing toggle constructs across the coxofemoral joints of small equids using a minimally invasive approach-A proof-of-concept cadaveric study.

OBJECTIVE To develop a minimally invasive technique for placing a toggle construct across the coxofemoral joint of small equids using computer-assisted surgery. STUDY DESIGN Experimental cadaveric study. SAMPLE POPULATION Three pilot specimens: One donkey, one Shetland pony and one Warmblood foal. Six main study specimens: Three Shetland ponies, one American Miniature Horse, one Warmblood foal and one donkey. METHODS Experimental surgeries were performed on both coxofemoral joints of each cadaver. Using a minimally invasive surgical approach, 5.5 mm bone canals were drilled through the femur and acetabulum, traversing the coxofemoral joint. Intraoperative guidance was provided by a cone-beam computed tomography (CBCT)-coupled surgical navigation system. A toggle construct was introduced through the bone canals. Surgical accuracy aberrations (SAA) were measured at the femoral entry and exit points and at the acetabular entry point on merged pre- and postoperative CBCT scans. The coxofemoral joint was assessed for articular cartilage damage by gross dissection. RESULTS A toggle construct was placed across all 18 coxofemoral joints. The overall median SAA in the main study was 2.8 mm (range: 0.4-8.0 mm). No cartilage damage was found in the cadaveric specimens of the main study. CONCLUSION The described technique allowed for the placement of a toggle construct across the coxofemoral joint of small equid cadaveric specimens without prior coxofemoral luxation. CLINICAL RELEVANCE This technique may serve as an option for surgical stabilization of coxofemoral joints in small equids. Further biomechanical investigations are required to assess optimal implant positioning and toggle constructs

Phosphocholine-Modified Lipooligosaccharides of Haemophilus influenzae Inhibit ATP-Induced IL-1beta Release by Pulmonary Epithelial Cells

Phosphocholine-modified bacterial cell wall components are virulence factors enabling immune evasion and permanent colonization of the mammalian host, by mechanisms that are poorly understood. Recently, we demonstrated that free phosphocholine (PC) and PC-modified lipooligosaccharides (PC-LOS) from Haemophilus influenzae, an opportunistic pathogen of the upper and lower airways, function as unconventional nicotinic agonists and efficiently inhibit the ATP-induced release of monocytic IL-1beta. We hypothesize that H. influenzae PC-LOS exert similar effects on pulmonary epithelial cells and on the complex lung tissue. The human lung carcinoma-derived epithelial cell lines A549 and Calu-3 were primed with lipopolysaccharide from Escherichia coli followed by stimulation with ATP in the presence or absence of PC or PC-LOS or LOS devoid of PC. The involvement of nicotinic acetylcholine receptors was tested using specific antagonists. We demonstrate that PC and PC-LOS efficiently inhibit ATP-mediated IL-1beta release by A549 and Calu-3 cells via nicotinic acetylcholine receptors containing subunits alpha7, alpha9, and/or alpha10. Primed precision-cut lung slices behaved similarly. We conclude that H. influenzae hijacked an endogenous anti-inflammatory cholinergic control mechanism of the lung to evade innate immune responses of the host. These findings may pave the way towards a host-centered antibiotic treatment of chronic airway infections with H. influenzae

Post-mortem computed tomographic angiography in equine distal forelimbs: A feasibility study

In-depth understanding of pathophysiological processes occurring in the vasculature of the equine distal limb is of great importance to improve both diagnostic and therapeutic approaches to diseases. To gain further insights, a model allowing high-resolution 3D-visualization of the vasculature is necessary. This pilot study evaluated the feasibility of restoring vascular perfusion in frozen-thawed distal equine cadaver limbs without prior preparation using computer tomographic imaging (CT). Five frozen-thawed, radiographically normal forelimbs were perfused with a lipophilic contrast agent through the median artery and radial vein in three phases (arterial, venous, and arterial-venous combined (AVC) dynamic). For comparison, one additional limb was perfused with a hydrosoluble contrast agent. The CT-studies (16-slice MDCT, 140 kV, 200 mA, 2 mm slice thickness, 1 mm increment, pitch 0.688) were evaluated at 11 specified regions for visualization of the vasculature and presence of artifacts or anatomic variations. The protocol used in this study proved to be feasible and provided good visualization (93.1%) of vasculature with low rates of artifacts. During the different phases, vascular visualization was similar, but while filling defects decreased in the later phases, extravasation worsened in the 2 limbs where it was observed. Subjectively, the best quality of angiographic images was achieved during the AVC dynamic phase. Perfusion with hydrosoluble contrast resulted in significantly lower vascular visualization (74.0%) and higher artifact rates. This study shows that reperfusion of frozen-thawed equine distal limbs with a lipophilic contrast agent allows for high-quality 3D-visualization of the vasculature and may serve as a model for in situ vascular evaluation in the future

Bekämpfung der Blutlaus (Eriosoma lanigerum Hausm.) in ökologischen Obstanlagen durch Etablierung von Ohrwürmern (Forficula auricularia L.) und Bewertung unterstützender Bekämpfungsverfahren

Das Forschungsprojekt „Bekämpfung der Blutlaus (Eriosoma lanigerum Hausm.) in ökologischen Obstanlagen durch Etablierung von Ohrwürmern (Forficula auricularia L.) und Bewertung unterstützender Bekämpfungsverfahren“ lief in Zusammenarbeit verschiedener Forschungs- und Beratungseinrichtungen von April 2007 bis Dezember 2009. Ziel des Projektes war es, auf Basis von Versuchsergebnissen eine betriebliche Behandlungsstrategie zur Bekämpfung der Blutlaus durch den gezielten Ohrwurmeinsatz und Ölbehandlungen zu entwickeln. Der Einfluss der Ohrwürmer erwies sich als schwer nachweisbar. Dies kann mehrere Gründe haben. Zum einen verteilen sich die Ohrwürmer über die Anlage, so dass ein Effekt in den einzelnen Varianten dadurch ausgeglichen wird, dass die Ohrwürmer nun auch in der Kontrolle zu finden waren. Zum anderen sind zum Zeitpunkt der Ansiedlung schon zahlreiche andere Nützlinge wie Marienkäfer und deren Larven, Schwebfliegen- und Florfliegenlarven sowie Blutlauszehrwespen in den Anlagen anzutreffen, die ihren Teil zur Eindämmung der Blutlauspopulation beitragen. Gerade das Massenhafte Auftreten des Asiatischen Marienkäfers (Harmonis axyridids) lässt eine Überlagerung des Nützlingseinflusses vermuten, da die Asiatischen Marienkäfer eine enorme Fraßleistung zeigen. Der Vergleich der Applikation von Öl mit dem Pinsel bzw. per Gebläsespritze zeigt bei einem hohen Befall deutliche Vorteile für das Pinseln. Die Behandlung ist direkter und wesentlich effektiver bei hohem Befall, allerdings auch Zeit aufwendig und hat keine Zulassung in Deutschland. Bei niedrigem Befallsdruck zeigte eine Frühjahrsspritzung durchaus zufrieden stellende Ergebnisse. Als Maßnahme bei akutem Befall kann eine Ansiedlung von Ohrwürmern also nicht dienen, eine langfristige Förderung ist aber sinnvoll. Bei Applikation oder Pinseln von Öl als Bekämpfungsmaßnahme im Frühjahr muss zwischen Befallsstärke und Arbeitszeitaufwand abgewogen werden. Durch Ohrwurmkot verschmutzte Äpfel treten durchaus auf, verursachten aber in keiner der Versuchsanlagen ökonomische Schäden. Ein Versuch zum Einfluss der mechanischen Bodenbearbeitung auf die Überwinternden Ohrwürmer zeigte keinen negativen Einfluss der Bodenbearbeitung auf die Ohrwurmpopulation. Es wird davon ausgegangen, dass die Tiere zwischen die Wurzeln der Pflanzen in den Fahrgassen ausweichen oder ihre Gelege so tief im Boden anlegen, dass sie nicht mehr von der Bodenbearbeitung beeinflusst werden. Die Untersuchungen zum Gesundheitszustand der Ohrwürmer ergaben den Nematoden Mermis nigrescens sowie Vertreter der Raupenfliegen (Tachinidae) als Hauptparasiten. Insgesamt lag der Anteil parasitierter oder kranker Tiere zwischen drei und acht Prozent