Infections with Avian Pathogenic and Fecal Escherichia coli Strains Display Similar Lung Histopathology and Macrophage Apoptosis

The purpose of this study was to compare histopathological changes in the lungs of chickens infected with avian pathogenic (APEC) and avian fecal (Afecal) Escherichia coli strains, and to analyze how the interaction of the bacteria with avian macrophages relates to the outcome of the infection. Chickens were infected intratracheally with three APEC strains, MT78, IMT5155, and UEL17, and one non-pathogenic Afecal strain, IMT5104. The pathogenicity of the strains was assessed by isolating bacteria from lungs, kidneys, and spleens at 24 h post-infection (p.i.). Lungs were examined for histopathological changes at 12, 18, and 24 h p.i. Serial lung sections were stained with hematoxylin and eosin (HE), terminal deoxynucleotidyl dUTP nick end labeling (TUNEL) for detection of apoptotic cells, and an anti-O2 antibody for detection of MT78 and IMT5155. UEL17 and IMT5104 did not cause systemic infections and the extents of lung colonization were two orders of magnitude lower than for the septicemic strains MT78 and IMT5155, yet all four strains caused the same extent of inflammation in the lungs. The inflammation was localized; there were some congested areas next to unaffected areas. Only the inflamed regions became labeled with anti-O2 antibody. TUNEL labeling revealed the presence of apoptotic cells at 12 h p.i in the inflamed regions only, and before any necrotic foci could be seen. The TUNEL-positive cells were very likely dying heterophils, as evidenced by the purulent inflammation. Some of the dying cells observed in avian lungs in situ may also be macrophages, since all four avian E. coli induced caspase 3/7 activation in monolayers of HD11 avian macrophages. In summary, both pathogenic and non-pathogenic fecal strains of avian E. coli produce focal infections in the avian lung, and these are accompanied by inflammation and cell death in the infected areas

The Medicine Tree: Unsettling palaeoecological perceptions of past environments and human activity

In this paper, we consider palaeoecological approaches to past landscapes and reflect upon how these are relevant to archaeological themes concerning concepts of environmental change and the role of past and present human communities in these processes. In particular, we highlight the importance of local context in the perception and understanding of landscape. Utilising a case study from Nepal, we look to ‘unsettle’ a conventional palaeoecological interpretation of a pollen record, originally constructed on western ecological principles, and instead draw on an interpretative perspective rooted in local Buddhist ecological knowledge, or a ‘folk taxonomy’, known as ‘The Medicine Tree’. We discuss how the interpretations of patterns and processes of vegetation change from a pollen record are not necessarily absolute. In particular, we outline how the palaeoecological frame of enquiry and reference is rooted in an essentially Eurocentric, Western scientific paradigm, which, in turn, shapes how we perceive and conceive of past landscapes and the role of ‘anthropogenic impact’ on vegetation. The aim of this is not to suggest that scientific approaches to the ‘reconstruction’ of past landscapes are necessarily invalid, but to illustrate how ‘empirical’ scientific methods and interpretations in archaeological science are contingent upon specific social and cultural frames of reference. We discuss the broader relevance of this, such as how we interpret past human activity and perception of landscape change, the ways in which we might look to mobilise research in the context of contemporary problems, issues concerning ‘degraded landscapes’ and how we incorporate local and archaeological perspectives with palaeoecology within an interconnected and iterative process

Human impact on the hydroenvironment of Lake Parishan, SW Iran, through the late Holocene

A multiproxy record from Lake Parishan, SW Iran, shows human impact on the lake and its catchment over the last 4000 years. The Parishan record provides evidence of changes in lake hydrology, from ostracod, diatom and isotope analyses, that are directly linked to human activity in the catchment; recorded by pollen and charcoal and supported by regional archaeological and historical data. The lake ostracod fauna is particularly sensitive to human induced catchment alterations and allow us to identify changes in catchment hydrology that are due to more than a simple change in precipitation: evaporation state. Oxygen isotope data from endogenic carbonates follow these faunal changes but also displays a longer trend to more positive values through the period, coincident with regional patterns of water balance for the late Holocene in the eastern Mediterranean

Triad3a induces the degradation of early necrosome to limit RipK1-dependent cytokine production and necroptosis.

Understanding the molecular signaling in programmed cell death is vital to a practical understanding of inflammation and immune cell function. Here we identify a previously unrecognized mechanism that functions to downregulate the necrosome, a central signaling complex involved in inflammation and necroptosis. We show that RipK1 associates with RipK3 in an early necrosome, independent of RipK3 phosphorylation and MLKL-induced necroptotic death. We find that formation of the early necrosome activates K48-ubiquitin-dependent proteasomal degradation of RipK1, Caspase-8, and other necrosomal proteins. Our results reveal that the E3-ubiquitin ligase Triad3a promotes this negative feedback loop independently of typical RipK1 ubiquitin editing enzymes, cIAPs, A20, or CYLD. Finally, we show that Triad3a-dependent necrosomal degradation limits necroptosis and production of inflammatory cytokines. These results reveal a new mechanism of shutting off necrosome signaling and may pave the way to new strategies for therapeutic manipulation of inflammatory responses

Inhibitor of apoptosis proteins, NAIP, cIAP1 and cIAP2 expression during macrophage differentiation and M1/M2 polarization

Monocytes and macrophages constitute the first line of defense of the immune system against external pathogens. Macrophages have a highly plastic phenotype depending on environmental conditions; the extremes of this phenotypic spectrum are a pro-inflammatory defensive role (M1 phenotype) and an anti-inflammatory tissue-repair one (M2 phenotype). The Inhibitor of Apoptosis (IAP) proteins have important roles in the regulation of several cellular processes, including innate and adaptive immunity. In this study we have analyzed the differential expression of the IAPs, NAIP, cIAP1 and cIAP2, during macrophage differentiation and polarization into M1 or M2. In polarized THP-1 cells and primary human macrophages, NAIP is abundantly expressed in M2 macrophages, while cIAP1 and cIAP2 show an inverse pattern of expression in polarized macrophages, with elevated expression levels of cIAP1 in M2 and cIAP2 preferentially expressed in M1. Interestingly, treatment with the IAP antagonist SMC-LCL161, induced the upregulation of NAIP in M2, the downregulation of cIAP1 in M1 and M2 and an induction of cIAP2 in M1 macrophages.This work was supported by Universidad de Granada, Plan Propio 2015;#P3B: FAM, VMC (http://investigacion.ugr.es/pages/planpropio/2015/ resoluciones/p3b_def_28072015); Universidad de Granada CEI BioTic;#CAEP2-84: VMC (http:// biotic.ugr.es/pages/resolucionprovisional enseaanzapractica22demayo/!); and Canadian nstitutes of Health Research;#231421, #318176, #361847: STB, ECL, RK (http://www.cihr-irsc.gc. ca/e/193.html). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Unique genome-wide transcriptome profiles of chicken macrophages exposed to Salmonella-derived endotoxin

<p>Abstract</p> <p>Background</p> <p>Macrophages play essential roles in both innate and adaptive immune responses. Bacteria require endotoxin, a complex lipopolysaccharide, for outer membrane permeability and the host interprets endotoxin as a signal to initiate an innate immune response. The focus of this study is kinetic and global transcriptional analysis of the chicken macrophage response to <it>in vitro </it>stimulation with endotoxin from <it>Salmonella </it><it>typhimurium</it>-798.</p> <p>Results</p> <p>The 38535-probeset Affymetrix GeneChip Chicken Genome array was used to profile transcriptional response to endotoxin 1, 2, 4, and 8 hours post stimulation (hps). Using a maximum FDR (False Discovery Rate) of 0.05 to declare genes as differentially expressed (DE), we found 13, 33, 1761 and 61 DE genes between endotoxin-stimulated versus non-stimulated cells at 1, 2, 4 and 8 hps, respectively. QPCR demonstrated that endotoxin exposure significantly affected the mRNA expression of <it>IL1B</it>, <it>IL6</it>, <it>IL8</it>, and <it>TLR15</it>, but not <it>IL10 </it>and <it>IFNG </it>in HD 11 cells. Ingenuity Pathway Analysis showed that 10% of the total DE genes were involved in inflammatory response. Three, 9.7, 96.8, and 11.8% of the total DE inflammatory response genes were significantly differentially expressed with endotoxin stimulation at 1, 2, 4 and 8 hps, respectively. The <it>NFKBIA, IL1B, IL8 and CCL4 </it>genes were consistently induced at all times after endotoxin treatment. <it>NLRC5 </it>(CARD domain containing, NOD-like receptor family, RCJMB04_18i2), an intracellular receptor, was induced in HD11 cells treated with endotoxin.</p> <p>Conclusions</p> <p>As above using an <it>in vitro </it>model of chicken response to endotoxin, our data revealed the kinetics of gene networks involved in host response to endotoxin and extend the known complexity of networks in chicken immune response to Gram-negative bacteria such as <it>Salmonella</it>. The induction of <it>NFKBIA, IL1B, IL8, CCL4 </it>genes is a consistent signature of host response to endotoxin over time. We make the first report of induction of a NOD-like receptor family member in response to <it>Salmonella </it>endotoxin in chicken macrophages.</p

Late-Holocene climatic variability south of the Alps as recorded by lake-level fluctuations at Lake Ledro, Trentino, Italy

International audienceA lake-level record for the late Holocene at Lake Ledro (Trentino, northeastern Italy) is presented. It is based on the sediment and pollen analysis of a 1.75 m high stratigraphic section observed on the southern shore (site Ledro I) and a 3.2 m long sediment core taken from a littoral mire on the southeastern shore (site Ledro II). The chronology is derived from 15 radiocarbon dates and pollen stratigraphy. The late-Holocene composite record established from these two sediment sequences gives evidence of centennial-scale fluctuations with highstands at c. 3400, 2600, 1700, 1200 and 400 cal. BP, in agreement with various palaeohydro-logical records established in central and northern Italy, as well as north of the Alps. In addition, high lake-level conditions at c. 2000 cal. BP may be the equivalent of stronger river discharge observed at the same time in Central Italy's rivers. In agreement with the lake-level record of Accesa (Tuscany), the Ledro record also suggests a relatively complex palaeohydrological pattern for the period around 4000 cal. BP. On a millennial scale, sediment hiatuses observed in the lower part of the Ledro I sediment sequence indicate that, except for a high-stand occurring just after 7500 cal. BP, lower lake levels generally prevailed rather before c. 4000 cal. BP than afterwards. Finally, the lake-level data obtained at Lake Ledro indicate that the relative continuity of settlements in humid areas of northern Italy during the Bronze Age (in contrast to their general abandonment north of the Alps between c. 3450 and 3150 cal. BP), does not reflect different regional patterns of climatic and palaeohy-drological conditions. In contrast, the rise in lake level dated to c. 3400 cal. BP at Ledro appears to coincide with a worldwide climate reversal, observed in both the hemispheres, while palaeoenvironmental and archaeological data collected at Lake Ledro may suggest, as a working hypothesis, a relative emancipation of proto-historic societies from climatic conditions

Modern pollen rain–vegetation relationships along a forest–steppe transect in the Golestan National Park, NE Iran

Pollen rain-vegetation relationships were studied over a forest-steppe transect in Golestan National Park, NE Iran. The surface pollen percentages were compared to the vegetation composition of the respective vegetation types in 18 sampling points using both descriptive and numerical approaches. Hyrcanian lowland forests are characterized by pollen assemblages dominated by Quercus, Carpinus betulus and low frequencies of Zelkova carpinifolia. Both Parrotia persica and Zelkova carpinifolia show a very low pollen representation in modern surface samples, an under-representation that should be taken into account in the interpretation of past vegetation records. Transitional communities between the forest and steppe including Acer monspessulanum subsp. turcomanicum, Crataegus and Paliurus scrubs, Juniperus excelsa woodlands and shrub-steppe patches are more difficult to distinguish in pollen assemblages, however, they are characterized by higher values of the dominant shrub species. The transitional vegetation communities at the immediate vicinity of the forest show also a substantial amount of grass pollen. Many insect-pollinated taxa are strongly under-represented in the pollen rain including most of the rosaceous trees and shrubs, Rhamnus, Paliurus, Acer and Berberis. Artemisia steppes are characterized by very high values of Artemisia pollen and the near absence of tree pollen

Paracrine cyclooxygenase-2 activity by macrophages drives colorectal adenoma progression in the Apc Min/+ mouse model of intestinal tumorigenesis

Genetic deletion or pharmacological inhibition of cyclooxygenase (COX)-2 abrogates intestinal adenoma development at early stages of colorectal carcinogenesis. COX-2 is localised to stromal cells (predominantly macrophages) in human and mouse intestinal adenomas. Therefore, we tested the hypothesis that paracrine Cox-2-mediated signalling from macrophages drives adenoma growth and progression in vivo in the ApcMin/+ mouse model of intestinal tumorigenesis. Using a transgenic C57Bl/6 mouse model of Cox-2 over-expression driven by the chicken lysozyme locus (cLys-Cox-2), which directs integration site-independent, copy number-dependent transgene expression restricted to macrophages, we demonstrated that stromal macrophage Cox-2 in colorectal (but not small intestinal) adenomas from cLys-Cox-2 x ApcMin/+ mice was associated with significantly increased tumour size (P = 0.025) and multiplicity (P = 0.025), compared with control ApcMin/+ mice. Transgenic macrophage Cox-2 expression was associated with increased dysplasia, epithelial cell Cox-2 expression and submucosal tumour invasion, as well as increased nuclear β-catenin translocation in dysplastic epithelial cells. In vitro studies confirmed that paracrine macrophage Cox-2 signalling drives catenin-related transcription in intestinal epithelial cells. Paracrine macrophage Cox-2 activity drives growth and progression of ApcMin/+ mouse colonic adenomas, linked to increased epithelial cell β-catenin dysregulation. Stromal cell (macrophage) gene regulation and signalling represent valid targets for chemoprevention of colorectal cancer