LacZ histochemistry and immunohistochemistry reveal ephrin-B ligand expression in the inner ear

Immunostaining in transgenic mice carrying the lac z gene can be used to map gene and protein distribution in a single tissue. In this study, we examined inner ears from ephrin-B3 homozygous and ephrin-B2 heterozygous mice. Ephrin-B3 lac z expression was limited in these mice. However, immunostaining revealed ephrin-B3 throughout cochlear and vestibular regions. Immunoreactivity was absent in ephrin-B3-homozygous null mutants, demonstrating the specificity of the antibody. Ephrin-B2 lac z reactivity was detected in a limited number of cells in cochlear and vestibular regions. Different immunostaining patterns were found with different antibodies. Comparison with lac z expression indicated which antibody was specific for the transmembrane-bound ephrin-B2 ligand

Extending the use of biologics to mucous membranes by attachment of a binding domain

Abstract Biologics are almost exclusively administered systemically, but localized delivery is preferable as it minimizes off-target exposure and allows more aggressive treatments. Topical application of biologics to epithelia is generally ineffective because most are covered with fluids and biologics are washed out too quickly to have significant therapeutic effects. Here we explore the idea that attaching a binding domain can serve as an “anchor” to extend the residency time of biologics on wet epithelia, allowing their effective use even with infrequent applications. We use topical application to the ocular surface as a challenging test since foreign substances are washed out especially efficiently by tear flow and blinking. Our results demonstrate that conjugation of antibodies to wheat germ agglutinin, which binds GlcNAc and sialic acid that are ubiquitously present in tissues, increases their half-life 350-fold upon application to the ocular surface in a mouse model of dry eye, a common and onerous disease in humans. Importantly, antibodies to IL-17A, IL-23, and IL-1β conjugated to the agglutinin reduces manifestations of dry eye, even when applied just once daily. In contrast, unconjugated antibodies are ineffective. Attaching an anchor to biologics is a simple means to overcome washout and to extend their therapeutic use

Construction and deflation of irrigation soils from the Pharaonic to the Roman period at Amheida (Trimithis), Da

Most of the environmental studies performed in the Western Desert of Egypt have dealt with climate change since the Late Pleistocene, notably with the humid Holocene (10000 to 4500 cal. BC) and following desertification. In the last dry period of the Holocene, human societies had to cope with changing environments and they selected refuges where water was permanently available. Some of the archaeological investigations in the Western Desert have studied the available water resources and the role of irrigation in the building of soils. In some cases, they have explained the abandonment of sites by the progressive depletion of water resources. This paper focuses on the site of Amheida (the Roman Trimithis) located in the Dakhla depression in the Western Desert of Egypt. It is an elongated area located south of a limestone plateau, where spring mounds provided abundant water during the period of occupation, which lasted from the Old Kingdom (2700 cal. BC) to the Late Roman period (end of the 41th century AD). Trimithis was abandoned after a long period of prosperity. The reasons for its abandonment, as well as that of other settlements in the oasis in the same period, have not been clarified yet. Geoarchaeological surveys carried out in 2011 and 2013 point to several phases including 1) the use of a spring mound during part of the Pharaonic period (2700 to 1069 cal. BC) and the formation of irrigation soils in the vicinity of spring mounds from the Late Pharaonic times (712 to 332 BC) to the beginning of the Roman period (30 BC); 2) the degradation of these soils by wind deflation in the irrigated perimeter during the early Roman period; 3) a recovery until the 3rd century AD, made possible by the digging of wells, before; 4) a severe phase of deflation and sand drifting which destroyed the irrigation soils and forced farmers to till the poorly irrigated bedrock.The Trimithis case exemplifies the way former societies could cope with harsh constraints (aridity, drifting sand and wind erosion) as long as water was plentiful. Water scarcity due to the progressive depletion of groundwater resources was probably compensated by an increase in human labour until thresholds were crossed, recovery being impossible due to water scarcity. A model summarizes the main findings of this study

Construction et destruction par déflation d’anthrosols des périodes pharaonique à romaine à Amheida (Ttimithis), oasis de Dakhla, désert occidental égyptien

International audienceMost of the environmental studies performed in the Western Desert of Egypt have dealt with climate change since the Late Pleistocene, notably with the humid Holocene (10000 to 4500 cal. BC) and following desertification. In the last dry period of the Holocene, human societies had to cope with changing environments and they selected refuges where water was permanently available. Some of the archaeological investigations in the Western Desert have studied the available water resources and the role of irrigation in the building of soils. In some cases, they have explained the abandonment of sites by the progressive depletion of water resources. This paper focuses on the site of Amheida (the Roman Trimithis) located in the Dakhla depression in the Western Desert of Egypt. It is an elongated area located south of a limestone plateau, where spring mounds provided abundant water during the period of occupation, which lasted from the Old Kingdom (2700 cal. BC) to the Late Roman period (end of the 4th century AD). Trimithis was abandoned after a long period of prosperity. The reasons for its abandonment, as well as that of other settlements in the oasis in the same period, have not been clarified yet.Geoarchaeological surveys carried out in 2011 and 2013 point to several phases including 1) the use of a spring mound during part of the Pharaonic period (2700 to 1069 cal. BC) and the formation of irrigation soils in the vicinity of spring mounds from the Late Pharaonic times (712 to 332 BC) to the beginning of the Roman period (30 BC); 2) the degradation of these soils by wind deflation in the irrigated perimeter during the early Roman period; 3) a recovery until the 3rd century AD, made possible by the digging of wells, before 4) a severe phase of deflation and sand drifting which destroyed the irrigation soils and forced farmers to till the poorly irrigated bedrock.The Trimithis case exemplifies the way former societies could cope with harsh constraints (aridity, drifting sand and wind erosion) as long as water was plentiful. Water scarcity due to the progressive depletion of groundwater resources was probably compensated by an increase in human labour until thresholds were crossed, recovery being impossible due to water scarcity. A model summarizes the main findings of this study.La plupart des études qui ont été réalisées dans le Désert occidental égyptien traitent du changement climatique depuis la fin du Pléistocène, notamment de l’Holocène humide (10 000-4500 cal. BC) et des modalités de la désertification. Pendant le deuxième Holocène aride, les sociétés humaines ont dû s’adapter à des environnements instables et choisir des refuges où l’eau était en permanence disponible. Une partie des recherches archéologiques réalisées dans le désert occidental ont pris en compte les ressources en eau disponibles et la place de l’irrigation dans la construction de certains sols (anthrosols). Elles ont aussi abordé la question de l’abandon de certains sites par l’épuisement progressif de la ressource en eau. Cet article se focalise sur Amheida (la Trimithis romaine) localisée dans la Dépression de Dakhla et qui fut occupée de la basse époque (712-332 BC) à la fin de la période romaine (fin du IVe siècle AD). Cette oasis est localisée au sud du plateau calcaire libyen ; des sources artésiennes ont fourni une eau abondante pendant la période d’occupation qui a duré de l’Ancien Empire (2700 cal. BC) à la fin de l’Empire romain (fin du IVe siècle AD). Trimithis a été abandonnée après une longue période de prospérité. Les raisons de l’abandon de ce site, comme d’autres dans la Dépression de Dakhla, n’ont pas encore été élucidées.Des recherches géoarchéologiques réalisées en 2011 et 2013 ont mis en évidence plusieurs phases : 1) l’utilisation de sources artésiennes pendant l’Ancien Empire (2700 à 1069 BC) et la formation d’anthrosols liée à l’irrigation de la Basse époque au début de la période romaine (30 BC) ; 2) la dégradation de ces sols par la déflation éolienne au début de la période romaine ; 3) une restauration partielle du périmètre irrigué avant la fin du IIIe siècle AD ; 4) une phase de déflation sévère qui a achevé l’érosion des sols et forcé les agriculteurs à cultiver sur le substrat schisteux.L’étude de Trimithis montre comment les sociétés antiques ont pu composer avec de fortes contraintes environnementales en milieu désertique, pour autant que l’eau était disponible en abondance. Cela s’est sans doute fait au prix d’un travail intense mais le déclin des ressources a empêché l’adaptation des sociétés et a conduit à un abandon rapide et inéluctable. Un modèle résume les principaux apports de cette étude

JP4-039, a Mitochondria-Targeted Nitroxide, Mitigates the Effect of Apoptosis and Inflammatory Cell Migration in the Irradiated Mouse Retina

We hypothesize that the injection of JP4-039, a mitochondria-targeted nitroxide, prior to irradiation of the mouse retina may decrease apoptosis and reduce neutrophil and macrophage migration into the retina. In our study, we aimed to examine the effects of JP4-039 in the mouse retina using fluorescent microscopy, a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay, and flow cytometry. Forty-five mice and one eye per mouse were used. In Group 1, fluorescent microscopy was used to determine retinal uptake of 10 µL (0.004 mg/µL) of intravitreally injected BODIPY-labeled JP4-039 at 0, 15, and 60 min after injection. In Group 2, the TUNEL assay was performed to investigate the rate of apoptosis after irradiation in addition to JP4-039 injection, compared to controls. In Group 3, flow cytometry was used to determine the extent of inflammatory cell migration into the retina after irradiation in addition to JP4-039 injection, compared to controls. Maximal retinal uptake of JP4-039 was 15 min after intravitreal injection (p p = 0.0066) and demonstrated reduced migration of N1 cells (30.7 ± 11.7% vs. 77.7 ± 5.3% controls; p = 0.004) and M1 cells (76.6 ± 4.2 vs. 88.1 ± 3.7% controls, p = 0.04). Pretreatment with intravitreally injected JP4-039 reduced apoptosis and inflammatory cell migration in the irradiated mouse retina, marking the first confirmed effect of this molecule in retinal tissue. Further studies may allow for safety profiling and potential use for patients with radiation retinopathy

Upregulation of keratocyte-specific gene expression in pellet cultures.

<p>Expression of six genes, previously identified as up-regulated during keratocyte differentiation, was determined after 2 weeks in pellet cultures derived from either MEM+FBS or N2 monolayer cultures as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056831#s2" target="_blank">Methods</a>. Gene expression is calculated relative to the NGFR+ derived hES cells. Error bars represent S.D. of triplicates. All genes were significantly (p<0.05) upregulated in pellets compared to NGFR+ cells except for CHST6. Asterisks show cases in which pellet culture induced a significant (p<0.05) increase in gene expression compared to the monolayers cultures.</p

Secretion of corneal keratan sulfate proteoglycans by pellet cultures.

<p>Proteoglycans were isolated from culture medium before (Lanes 1,2,5,6) or after (3,4,7,8) three-week incubation with hES pellets. The proteoglycan fractions were biotin- labeled and immune-precipitated with antibodies against keratocan (anti-Kera) (lanes 1–4) or keratan sulfate glycosaminoglycan (anti-KS) (lanes 5–8). Half of each sample was digested with endo-ß-galactosidase (as described under <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056831#s2" target="_blank">Methods</a>) to hydrolyze keratan sulfate, and samples were separated by SDS-PAGE, transferred to PVDF membranes and biotinylated proteins detected with avidin-labeled infrared dye as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056831#s2" target="_blank">Methods</a>. Presence of biotinylated proteins migrating as a broad, heterogeneous >100 band typical of keratan sulfate proteoglycan (KSPG - bracket on the left) was present in lanes 3 and 7. Sensitivity of this material to digestion with keratan sulfate-specific glycosidase (lanes 4 and 8) demonstrates presence of keratocan-linked keratan sulfate, a unique keratocyte biosynthetic product.</p

TERRANOVA from the last and current Interglacial periods into the Anthropocene: an Atlas database drawing lessons from ancient land use for future European landscape management

International audienceTERRANOVA is a Marie Skłodowska-Curie Innovative Training Networks (H2020-MSCA-ITN) project (2019-2023) training 15 PhD students in a new learning initiative between Humanities and Science: Mapping past environments and energy regimes, rethinking human-environment interaction and designing land management tools for policy. TERRANOVA will produce an unprecedented atlas with layers of reconstructed and modelled land-use and vegetation dynamics, climate change and mega-fauna history in Europe from the Eemian (Last Interglacial) and the Holocene from the start up until the present day. This paper describes the intermediate results of two years of research into Atlas building. Communication and data exchange, as well as the process of atlas generation work flow, have been undertaken, including examples of datasets from deep history, ancient landscapes, energy regimes and climate scenarios. The atlas database implements state-of-the-art standards for increasing the interoperability of spatiotemporal datasets. It is currently formed by four main data types: Archaeological data, Climate data, Land cover data, and Megafauna (i.e. large mammals) distribution. The intermediate publication concludes with listing the next steps to stream the Terranova atlas as a tool for communicating the European history of environmental change, including support for future landscape management policies

Co-culture with PA6 cells induces upregulation of neural crest gene expression in hES cells.

<p>hES cells were co-cultured with PA6 as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056831#s2" target="_blank">Methods</a>. RNA was isolated at post-induction days (PID) 2, 4, 6, and 8. Expression of characteristic neural crest (NC) marker genes was determined by qPCR as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056831#s2" target="_blank">Methods</a> using human-specific primers (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0056831#pone-0056831-t002" target="_blank">Table 2</a>). Expression levels are calculated relative to untreated hES cells (hES = 1). Error bars show the standard deviation (S.D.) of triplicate analyses.</p