Basic science232. Certolizumab pegol prevents pro-inflammatory alterations in endothelial cell function

Background: Cardiovascular disease is a major comorbidity of rheumatoid arthritis (RA) and a leading cause of death. Chronic systemic inflammation involving tumour necrosis factor alpha (TNF) could contribute to endothelial activation and atherogenesis. A number of anti-TNF therapies are in current use for the treatment of RA, including certolizumab pegol (CZP), (Cimzia ®; UCB, Belgium). Anti-TNF therapy has been associated with reduced clinical cardiovascular disease risk and ameliorated vascular function in RA patients. However, the specific effects of TNF inhibitors on endothelial cell function are largely unknown. Our aim was to investigate the mechanisms underpinning CZP effects on TNF-activated human endothelial cells. Methods: Human aortic endothelial cells (HAoECs) were cultured in vitro and exposed to a) TNF alone, b) TNF plus CZP, or c) neither agent. Microarray analysis was used to examine the transcriptional profile of cells treated for 6 hrs and quantitative polymerase chain reaction (qPCR) analysed gene expression at 1, 3, 6 and 24 hrs. NF-κB localization and IκB degradation were investigated using immunocytochemistry, high content analysis and western blotting. Flow cytometry was conducted to detect microparticle release from HAoECs. Results: Transcriptional profiling revealed that while TNF alone had strong effects on endothelial gene expression, TNF and CZP in combination produced a global gene expression pattern similar to untreated control. The two most highly up-regulated genes in response to TNF treatment were adhesion molecules E-selectin and VCAM-1 (q 0.2 compared to control; p > 0.05 compared to TNF alone). The NF-κB pathway was confirmed as a downstream target of TNF-induced HAoEC activation, via nuclear translocation of NF-κB and degradation of IκB, effects which were abolished by treatment with CZP. In addition, flow cytometry detected an increased production of endothelial microparticles in TNF-activated HAoECs, which was prevented by treatment with CZP. Conclusions: We have found at a cellular level that a clinically available TNF inhibitor, CZP reduces the expression of adhesion molecule expression, and prevents TNF-induced activation of the NF-κB pathway. Furthermore, CZP prevents the production of microparticles by activated endothelial cells. This could be central to the prevention of inflammatory environments underlying these conditions and measurement of microparticles has potential as a novel prognostic marker for future cardiovascular events in this patient group. Disclosure statement: Y.A. received a research grant from UCB. I.B. received a research grant from UCB. S.H. received a research grant from UCB. All other authors have declared no conflicts of interes

Stable population structure in Europe since the Iron Age, despite high mobility

Ancient DNA research in the past decade has revealed that European population structure changed dramatically in the prehistoric period (14,000–3000 years before present, YBP), reflecting the widespread introduction of Neolithic farmer and Bronze Age Steppe ancestries. However, little is known about how population structure changed from the historical period onward (3000 YBP - present). To address this, we collected whole genomes from 204 individuals from Europe and the Mediterranean, many of which are the first historical period genomes from their region (e.g. Armenia and France). We found that most regions show remarkable inter-individual heterogeneity. At least 7% of historical individuals carry ancestry uncommon in the region where they were sampled, some indicating cross-Mediterranean contacts. Despite this high level of mobility, overall population structure across western Eurasia is relatively stable through the historical period up to the present, mirroring geography. We show that, under standard population genetics models with local panmixia, the observed level of dispersal would lead to a collapse of population structure. Persistent population structure thus suggests a lower effective migration rate than indicated by the observed dispersal. We hypothesize that this phenomenon can be explained by extensive transient dispersal arising from drastically improved transportation networks and the Roman Empire’s mobilization of people for trade, labor, and military. This work highlights the utility of ancient DNA in elucidating finer scale human population dynamics in recent history

Genomic investigations of unexplained acute hepatitis in children

Since its first identification in Scotland, over 1,000 cases of unexplained paediatric hepatitis in children have been reported worldwide, including 278 cases in the UK1. Here we report an investigation of 38 cases, 66 age-matched immunocompetent controls and 21 immunocompromised comparator participants, using a combination of genomic, transcriptomic, proteomic and immunohistochemical methods. We detected high levels of adeno-associated virus 2 (AAV2) DNA in the liver, blood, plasma or stool from 27 of 28 cases. We found low levels of adenovirus (HAdV) and human herpesvirus 6B (HHV-6B) in 23 of 31 and 16 of 23, respectively, of the cases tested. By contrast, AAV2 was infrequently detected and at low titre in the blood or the liver from control children with HAdV, even when profoundly immunosuppressed. AAV2, HAdV and HHV-6 phylogeny excluded the emergence of novel strains in cases. Histological analyses of explanted livers showed enrichment for T cells and B lineage cells. Proteomic comparison of liver tissue from cases and healthy controls identified increased expression of HLA class 2, immunoglobulin variable regions and complement proteins. HAdV and AAV2 proteins were not detected in the livers. Instead, we identified AAV2 DNA complexes reflecting both HAdV-mediated and HHV-6B-mediated replication. We hypothesize that high levels of abnormal AAV2 replication products aided by HAdV and, in severe cases, HHV-6B may have triggered immune-mediated hepatic disease in genetically and immunologically predisposed children

Distinct positions of genetic and oral histories: Perspectives from India

Summary: Over the past decade, genomic data have contributed to several insights on global human population histories. These studies have been met both with interest and critically, particularly by populations with oral histories that are records of their past and often reference their origins. While several studies have reported concordance between oral and genetic histories, there is potential for tension that may stem from genetic histories being prioritized or used to confirm community-based knowledge and ethnography, especially if they differ. To investigate the interplay between oral and genetic histories, we focused on the southwestern region of India and analyzed whole-genome sequence data from 156 individuals identifying as Bunt, Kodava, Nair, and Kapla. We supplemented limited anthropological records on these populations with oral history accounts from community members and historical literature, focusing on references to non-local origins such as the ancient Scythians in the case of Bunt, Kodava, and Nair, members of Alexander the Great’s army for the Kodava, and an African-related source for Kapla. We found these populations to be genetically most similar to other Indian populations, with the Kapla more similar to South Indian tribal populations that maximize a genetic ancestry related to Ancient Ancestral South Indians. We did not find evidence of additional genetic sources in the study populations than those known to have contributed to many other present-day South Asian populations. Our results demonstrate that oral and genetic histories may not always provide consistent accounts of population origins and motivate further community-engaged, multi-disciplinary investigations of non-local origin stories in these communities

Stable population structure in Europe since the Iron Age, despite high mobility

Ancient DNA research in the past decade has revealed that European population structure changed dramatically in the prehistoric period (14,000-3000 years before present, YBP), reflecting the widespread introduction of Neolithic farmer and Bronze Age Steppe ancestries. However, little is known about how population structure changed from the historical period onward (3000 YBP - present). To address this, we collected whole genomes from 204 individuals from Europe and the Mediterranean, many of which are the first historical period genomes from their region (e.g. Armenia and France). We found that most regions show remarkable inter-individual heterogeneity. At least 7% of historical individuals carry ancestry uncommon in the region where they were sampled, some indicating cross-Mediterranean contacts. Despite this high level of mobility, overall population structure across western Eurasia is relatively stable through the historical period up to the present, mirroring geography. We show that, under standard population genetics models with local panmixia, the observed level of dispersal would lead to a collapse of population structure. Persistent population structure thus suggests a lower effective migration rate than indicated by the observed dispersal. We hypothesize that this phenomenon can be explained by extensive transient dispersal arising from drastically improved transportation networks and the Roman Empire's mobilization of people for trade, labor, and military. This work highlights the utility of ancient DNA in elucidating finer scale human population dynamics in recent history

A genetic history of continuity and mobility in the Iron Age central Mediterranean

The Iron Age was a dynamic period in central Mediterranean history, with the expansion of Greek and Phoenician colonies and the growth of Carthage into the dominant maritime power of the Mediterranean. These events were facilitated by the ease of long-distance travel following major advances in seafaring. We know from the archaeological record that trade goods and materials were moving across great distances in unprecedented quantities, but it is unclear how these patterns correlate with human mobility. Here, to investigate population mobility and interactions directly, we sequenced the genomes of 30 ancient individuals from coastal cities around the central Mediterranean, in Tunisia, Sardinia and central Italy. We observe a meaningful contribution of autochthonous populations, as well as highly heterogeneous ancestry including many individuals with non-local ancestries from other parts of the Mediterranean region. These results highlight both the role of local populations and the extreme interconnectedness of populations in the Iron Age Mediterranean. By studying these trans-Mediterranean neighbours together, we explore the complex interplay between local continuity and mobility that shaped the Iron Age societies of the central Mediterranean

Early Holocene cemetery, burials and skulls.

<p>(A)-Gobero site G3 showing excavated burials (red dots). (B)-Enlarged map of the early Holocene cemetery showing the location of 17 undisturbed burials of skeletons with dark-stained bone (red dots). Five burials (red dot with outer ring) were directly dated to a narrow range of ∼7500±250 years B.C.E. (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002995#pone-0002995-g002" target="_blank">Figure 2</a>; <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002995#pone-0002995-t002" target="_blank">Table 2</a>). (C)-Skeleton (dark-stained) of an early Holocene adult male (G3B8; ∼7515 B.C.E.) buried in supine, hyperflexed posture with hands over the mouth and feet crossed. Computed-tomography cross-section (below) across the middle of the skeleton (red line) shows the tightly bundled configuration of major limb bones (within a 25 cm×12 cm rectangle) for an adult with stature approximately 2 m. (D)-Skull of early Holocene adult male (as in C) showing long, low calvarium, broad zygomatic width and relatively flat face. (E)-Skull of an early Holocene juvenile (G3B17b; ∼7630 B.C.E; estimated age 5 years) already showing long, low cranial proportions. Scale bar in C equals 13.3 cm for skeleton and 10 cm for CT scan; skull length (glabella-opisthocranion) in D and E equals 190.0 mm and 171.0 mm, respectively. <i>Abbreviations</i>: <i>f</i>, femur, <i>fi</i>, fibula; <i>h</i>, humerus; <i>r</i>, radius; <i>ti</i>, tibia; <i>ul</i>, ulna.</p

Principal components analysis of craniofacial dimensions among Late Pleistocene to mid-Holocene populations from the Maghreb and southern Sahara.

<p>Plot of first two principal components extracted from a mean matrix for 17 craniometric variables (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002995#pone-0002995-t004" target="_blank">Tables 4</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002995#pone-0002995-t007" target="_blank">7</a>) in 9 human populations (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002995#pone-0002995-t003" target="_blank">Table 3</a>) from the Late Pleistocene through the mid-Holocene from the Maghreb and southern Sahara. Seven trans-Saharan populations cluster together, whereas Late Pleistocene Aterians (Ater) and the mid-Holocene population at Gobero (Gob-m) are striking outliers. Axes are scaled by the square root of the corresponding eigenvalue for the principal component. <i>Abbreviations</i>: <i>Ater</i>, Aterian; <i>EMC</i>, eastern Maghreb Capsian; <i>EMI</i>, eastern Maghreb Iberomaurusian; <i>Gob-e</i>, Gobero early Holocene; <i>Gob-m</i>, Gobero mid-Holocene; <i>Mali</i>, Hassi-el-Abiod, Mali; <i>Maur</i>, Mauritania; <i>WMC</i>, western Maghreb Capsian; <i>WMI</i>, western Maghreb Iberomaurusian.</p

Craniometric means for human samples.

<p>Craniometric means for early and mid-Holocene skulls from Gobero and comparative samples elsewhere from northern Africa. <i>Measurement acronyms</i> (after <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002995#pone.0002995-Dutour1" target="_blank">[18]</a>): <i>LGO</i>, glabella-occipital length; <i>BPX</i>, maximum cranial breadth; <i>LBN</i>, basion-nasion; <i>HBB</i>, basion-bregma; <i>AFR</i>, frontal arc; <i>APA</i>, parietal arc; <i>AOC</i>, occipital arc; <i>CFR</i>, frontal chord; <i>COC</i>, occipital chord; <i>HNP</i>, nasion-prosthion; <i>HNZ</i>, nasion-nasospinale; <i>BNZ</i>, nasal breadth; <i>BZY</i>, bizygomtic breadth; <i>BFW</i>, minimum frontal breadth; <i>BFX</i>, maximum frontal breadth; <i>HORC</i>, horizontal circumference above the superciliary arches. <i>Sample acronyms and abbreviations</i>: <i>Ater</i>, Aterian; <i>EMI</i>, eastern Maghreb Iberomaurusian; <i>EMC</i>, eastern Maghreb Capsian; <i>Gob-e</i>, Gobero early Holocene; <i>Gob-m</i>, Gobero mid-Holocene; <i>Maur</i>, Mauritania; <i>Mali</i>, Hassi-el-Abiod, Mali; <i>WMI</i>, western Maghreb Iberomaurusian; <i>WMC</i>, western Maghreb Capsian.</p

Mid-Holocene midden.

<p>Portion of a mid-Holocene midden (midden 4) with matrix removed showing stacking of the valves of the clam Mutela, articulated fish vertebrae, and potsherds (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0002995#pone-0002995-t002" target="_blank">Table 2, dates 42, 43</a>, average midpoint ∼4445 B.C.E.).</p