The earliest unambiguous Neanderthal engravings on cave walls: La Roche-Cotard, Loire Valley, France.

Here we report on Neanderthal engravings on a cave wall at La Roche-Cotard (LRC) in central France, made more than 57±3 thousand years ago. Following human occupation, the cave was completely sealed by cold-period sediments, which prevented access until its discovery in the 19th century and first excavation in the early 20th century. The timing of the closure of the cave is based on 50 optically stimulated luminescence ages derived from sediment collected inside and from around the cave. The anthropogenic origin of the spatially-structured, non-figurative marks found within the cave is confirmed using taphonomic, traceological and experimental evidence. Cave closure occurred significantly before the regional arrival of H. sapiens, and all artefacts from within the cave are typical Mousterian lithics; in Western Europe these are uniquely attributed to H. neanderthalensis. We conclude that the LRC engravings are unambiguous examples of Neanderthal abstract design.

Mechanically Enhanced Salmo salar Gelatin by Enzymatic Cross-linking: Premise of a Bioinspired Material for Food Packaging, Cosmetics, and Biomedical Applications.

Marine animal by-products of the food industry are a great source of valuable biomolecules. Skins and bones are rich in collagen, a protein with various applications in food, cosmetic, healthcare, and medical industries in its native form or partially hydrolyzed (gelatin). Salmon gelatin is a candidate of interest due to its high biomass production available through salmon consumption, its biodegradability, and its high biocompatibility. However, its low mechanical and thermal properties can be an obstacle for various applications requiring cohesive material. Thus, gelatin modification by cross-linking is necessary. Enzymatic cross-linking by microbial transglutaminase (MTG) is preferred to chemical cross-linking to avoid the formation of potentially cytotoxic residues. In this work, the potential of salmon skin gelatin was investigated, in a comparative study with porcine gelatin, and an enzymatic versus chemical cross-linking analysis. For this purpose, the two cross-linking methods were applied to produce three-dimensional, porous, and mechanically reinforced hydrogels and sponges with different MTG ratios (2%, 5%, and 10% w/w gelatin). Their biochemical, rheological, and structural properties were characterized, as well as the stability of the material, including the degree of syneresis and the water-binding capacity. The results showed that gelatin enzymatically cross-linked produced material with high cross-linking densities over 70% of free amines. The MTG addition seemed to play a crucial role, as shown by the increase in mechanical and thermal resistances with the production of a cohesive material stable above 40 °C for at least 7 days and comparable to porcine and chemically cross-linked gelatins. Two prototypes were obtained with similar thermal resistances but different microstructures and viscoelastic properties, due to different formation dynamics of the covalent network. Considering these results, the enzymatically cross-linked salmon gelatin is a relevant candidate as a biopolymer for the production of matrix for a wide range of biotechnological applications such as food packaging, cosmetic patch, wound healing dressing, or tissue substitute.

Apparent Yield Stress of Sputum as a Relevant Biomarker in Cystic Fibrosis.

The mucus obstructing the airways of Cystic Fibrosis (CF) patients is a yield stress fluid. Linear and non-linear rheological analyses of CF sputa can provide relevant biophysical markers, which could be used for the management of this disease. Sputa were collected from CF patients either without any induction or following an aerosol treatment with the recombinant human DNAse (rhDNAse, Pulmozyme®). Several sample preparations were considered and multiple measurements were performed in order to assess both the repeatability and the robustness of the rheological measurements. The linear and non-linear rheological properties of all CF sputa were characterized. While no correlation between oscillatory shear linear viscoelastic properties and clinical data was observed, the steady shear flow data showed that the apparent yield stress of sputum from CF patients previously treated with rhDNAse was approximately one decade lower than that of non-treated CF patients. Similar results were obtained with sputa from non-induced CF patients subjected ex vivo to a Pulmozyme® aerosol treatment. The results demonstrate that the apparent yield stress of patient sputa is a relevant predictive/prognostic biomarker in CF patients and could help in the development of new mucolytic agents.

Nanocellulose-based foam morphological, mechanical and thermal properties in relation to hydrogel precursor structure and rheology.

Foams were prepared from nanocellulose-based hydrogel precursors using a freeze-drying process. The work mainly aims at investigating the relationships between the mechanical and thermal properties of foams and the rheological properties of their hydrogel precursors, which were characterized in a previous paper. The structure of foams was characterized by SEM and confocal microscopy, their elasticity by compression tests, and their thermal conductivity by hot strip as well as transient pulsed techniques. A strong correlation was shown between the elastic properties of foams and those of their hydrogel precursors, and a minimum thermal conductivity was shown to appear at a cellulose volume fraction corresponding to a transition in viscoelastic properties of hydrogels. Results suggest that foams and hydrogels share common microstructural features, which makes it possible to tune the mechanical and thermal properties of foams by tuning the rheological properties of their hydrogel precursors.

Optimizations of In Vitro Mucus and Cell Culture Models to Better Predict In Vivo Gene Transfer in Pathological Lung Respiratory Airways: Cystic Fibrosis as an Example.

The respiratory epithelium can be affected by many diseases that could be treated using aerosol gene therapy. Among these, cystic fibrosis (CF) is a lethal inherited disease characterized by airways complications, which determine the life expectancy and the effectiveness of aerosolized treatments. Beside evaluations performed under in vivo settings, cell culture models mimicking in vivo pathophysiological conditions can provide complementary insights into the potential of gene transfer strategies. Such models must consider multiple parameters, following the rationale that proper gene transfer evaluations depend on whether they are performed under experimental conditions close to pathophysiological settings. In addition, the mucus layer, which covers the epithelial cells, constitutes a physical barrier for gene delivery, especially in diseases such as CF. Artificial mucus models featuring physical and biological properties similar to CF mucus allow determining the ability of gene transfer systems to effectively reach the underlying epithelium. In this review, we describe mucus and cellular models relevant for CF aerosol gene therapy, with a particular emphasis on mucus rheology. We strongly believe that combining multiple pathophysiological features in single complex cell culture models could help bridge the gaps between in vitro and in vivo settings, as well as viral and non-viral gene delivery strategies.

Structure and rheology of aqueous suspensions and hydrogels of cellulose nanofibrils: Effect of volume fraction and ionic strength.

The structure and rheology of TEMPO-oxidized cellulose nanofibrils (CNF) suspensions and hydrogels, used as precursors in the elaboration of bio-based aerogels for thermal insulation applications, were studied as a function of CNF volume fraction and ionic strength. The CNF geometry and rigidity were evaluated using AFM observations. Viscometric measurements, performed at very low CNF concentrations, highlighted the prominent role played by electroviscous effects, which can be modulated by ionic strength. Oscillatory measurements on semi-dilute CNF suspensions revealed the formation of a three-dimensional hydrogel network above a percolation fraction, which was shown to depend on the ionic strength. The rheological properties of CNF hydrogels were shown to depend on CNF fraction and ionic strength. In deionized water, the existence of two different concentration regimes was discussed in terms of network structural characteristics and CNF interactions.

Precise dating of the Middle-to-Upper Paleolithic transition in Murcia (Spain) supports late Neandertal persistence in Iberia.

The late persistence in Southern Iberia of a Neandertal-associated Middle Paleolithic is supported by the archeological stratigraphy and the radiocarbon and luminescence dating of three newly excavated localities in the Mula basin of Murcia (Spain). At Cueva Antón, Mousterian layer I-k can be no more than 37,100 years-old. At La Boja, the basal Aurignacian can be no less than 36,500 years-old. The regional Middle-to-Upper Paleolithic transition process is thereby bounded to the first half of the 37th millennium Before Present, in agreement with evidence from Andalusia, Gibraltar and Portugal. This chronology represents a lag of minimally 3000 years with the rest of Europe, where that transition and the associated process of Neandertal/modern human admixture took place between 40,000 and 42,000 years ago. The lag implies the presence of an effective barrier to migration and diffusion across the Ebro river depression, which, based on available paleoenvironmental indicators, would at that time have represented a major biogeographical divide. In addition, (a) the Phlegraean Fields caldera explosion, which occurred 39,850 years ago, would have stalled the Neandertal/modern human admixture front because of the population sink it generated in Central and Eastern Europe, and (b) the long period of ameliorated climate that came soon after (Greenland Interstadial 8, during which forests underwent a marked expansion in Iberian regions south of 40°N) would have enhanced the "Ebro Frontier" effect. These findings have two broader paleoanthropological implications: firstly, that, below the Ebro, the archeological record made prior to 37,000 years ago must be attributed, in all its aspects and components, to the Neandertals (or their ancestors); secondly, that modern human emergence is best seen as an uneven, punctuated process during which long-lasting barriers to gene flow and cultural diffusion could have existed across rather short distances, with attendant consequences for ancient genetics and models of human population history.

A non-hypocholesterolemic atorvastatin treatment improves vessel elasticity by acting on elastin composition in WHHL rabbits.

BACKGROUND AND AIMS: Statins are prescribed for their preventative effects within atherosclerosis development. To our knowledge, no study focusing on very low-dose (non-hypolipidemic effect) and long-term atorvastatin treatment in vivo was available. Our aim was to assess the effect of such atorvastatin treatment on the mechanical and functional characteristics of arteries in the context of primary prevention. METHODS: An atorvastatin treatment (2.5 mg/kg/day) was tested against controls on 34 male 3 to 12 month-old WHHL rabbits. No effect on total cholesterol, triglycerides, HDL or LDL was observed. The arterial stiffness was evaluated on vigil animals by pulse wave velocity (PWV) measurement. Then, in vitro measurements were made to evaluate (1) the endothelial and vascular smooth muscle function, (2) the elasticity of the arterial wall and (3) the composition in collagen and elastin in the aorta. RESULTS: The PWV increasing observed with age in control group was canceled by treatment, creating a significance difference between groups at 12 months (5.17 ± 0.50 vs 2.14 ± 0.34 m s(-1) in control and treated groups respectively). Vasoreactivity modifications can't explain this result but maintain of elasticity with treatment in large arteries was confirm by a static tensile test. A first possible explanation is the change of wall composition with treatment, validated by the percentage of elastin at 12 months, 4.4% lower in the control group compared to the treated group (p < 0.05). CONCLUSIONS: This study shows that a non-hypocholesterolemic statin treatment could improve vessel elasticity in the atherosclerotic WHHL model. The great novelty of this work is the vessel wall composition changing associated. This first approach in animal opens the reflection on the use of these low doses in humans. This could be interesting in the context of arterial stiffening with aging, non-hyperlipidemic atherosclerosis or with cholesterol reduce by another therapy or lifestyle modification.

Evaluation of low viscosity variations in fluids using temporal and spatial analysis of the speckle pattern.

The noninvasive detection of a material's viscoelasticity is of great importance in the medical field. In fact, certain diseases cause changes in tissue structure and biological fluid viscosity; tracking those changes allows for detection of these diseases. Rheological measurements are also imperative in the industrial field, where it is necessary to characterize a material's viscoelasticity for manufacturing purposes. In this Letter, we present a noncontact, noninvasive, and low cost method for determining low viscosity values and variations in fluids. Laser speckle and viscometric measurements are performed on test samples having low scattering coefficients and low viscosities. The speckle spatial analysis proved to be as accurate as the speckle temporal correlation method reported in previous studies. Very low viscosities of the order of 1 mPa.s were retrieved for the first time using speckle images with either a frame rate of 1950 fps or a single acquired image.

Rheological study of reinforcement of agarose hydrogels by cellulose nanowhiskers.

The influence of the addition of tunicate cellulose nanowhiskers on the structural and rheological properties of an agarose hydrogel matrix has been studied, with the objective to design innovative green material, with good mechanical properties. The cellulose nanowhiskers were characterized using transmission electron microscopy, and their charge surface density was determined by a titration method. Oscillatory shear and stress relaxation tests were performed in order to characterize the rheological properties of the agarose matrix, and of the agarose hydrogels filled by nanowhiskers at volume fractions below 0.2%. The results show a significant reinforcement effect due to the addition of nanowhiskers, and suggest changes in the matrix network structure induced by the cellulose nanoparticles.

A multi-scale approach of the mechanisms underlying exopolysaccharide auto-organization in the Proteus mirabilis extracellular matrix.

For decades, the origin of the concentric ring pattern of bacterial swarming colonies has puzzled microbiologists. It was hypothesized that a periodic water activity variation originates a phase transition within the extracellular matrix water H bond network, which switches on and off the exopolysaccharide auto-organization. Both rheological and infrared spectroscopy measurements respectively performed at a molecular scale and on a currently migrating colony, have given a physical insight into the mechanisms which underlie the switch between swarming and consolidation phases. Thanks to in situ and real time infrared microspectroscopy, and thanks to the brilliance of the infrared beam at SOLEIL synchrotron, here we demonstrate that Proteus mirabilis swarming is triggered by a periodic variation of water activity at the colony edge. A dynamic behavior emerges from the global properties of the multicellular entity which relies on the ability of the bacterial cells to tune exoproduct synthesis in order to undergo sharp transitions at a given water activity threshold.

Linear rheological properties of low molecular weight polyethylene glycol solutions.

This experimental study of the linear viscoelastic properties of PEG600 aqueous solutions at various concentrations and temperatures mainly aims at getting a better understanding, in concentrated regimes, of the role played on the structural and rheological properties by PEG assembly properties previously evidenced in the dilute regime. The results indicate a peculiar and unique viscoelastic behavior: the elastic modulus versus frequency curves are nearly the same for all concentrations and temperatures investigated. The key role played by hydrogen bonds in the rheological properties of PEG solutions is highlighted. The relaxation on long time scales is indicative of large scale complex associative polymeric structures. A schematic complete phase diagram of PEG600 in water is finally proposed.

Stratigraphic and technological evidence from the Middle Palaeolithic-Châtelperronian-Aurignacian record at the Bordes-Fitte rockshelter (Roches d'Abilly site, Central France).

This paper presents a geoarchaeological study of Middle and Upper Palaeolithic (Châtelperronian, Aurignacian and Solutrean) occupations preserved at the Bordes-Fitte rockshelter in Central France. The lithostratigraphic sequence is composed of near-surface sedimentary facies with vertical and lateral variations, in a context dominated by run-off and gravitational sedimentary processes. Field description and micromorphological analysis permit us to reconstruct several episodes of sediment slope-wash and endokarst dynamics, with hiatuses and erosional phases. The archaeostratigraphic succession includes Châtelperronian artefacts, inter-stratified between Middle Palaeolithic and Aurignacian occupations. Systematic refitting and spatial analysis reveal that the Châtelperronian point production and flake blanks retouched into denticulates, all recovered in the same stratigraphic unit, result from distinct and successive occupations and are not a 'transitional' Middle to Upper Palaeolithic assemblage. The ages obtained by (14)C place the Châtelperronian occupation in the 41-48 ka cal BP (calibrated thousands of years before present) interval and are consistent with the quartz optically stimulated luminescence age of 39 ± 2 ka and feldspar infra-red stimulated luminescence age of 45 ± 2 ka of the sediments. The Bordes-Fitte rockshelter sequence represents an important contribution to the debate about the characterization and timing of the Châtelperronian, as well as its affinities to earlier and later industries.

Does water activity rule P. mirabilis periodic swarming? I. Biochemical and functional properties of the extracellular matrix.

The dynamics of bacterial colonies is complex in nature because it correlates the behavior of numerous individual cells in space and time and is characterized by emergent properties such as virulence or antibiotics resistance. Because there is no clear-cut evidence that periodic swarming of P. mirabilis colonies is ruled by chemical triggers responsible for cell-to-cell signaling in most of the biofilms, we propose that the observed periodicity relies on the colony's global properties. Hence, the biochemical and functional properties of the extracellular matrix (ECM) of P. mirabilis colonies were investigated. A binary exopolysaccharide mixture (1 and 300 kDa), glycinebetaine, and a phenoglycolipid were identified. Rheology, calorimetry, and water sorption experiments performed on purified EPS bring evidence that these exoproducts exhibit marked viscoelasticity, which likely relies on large scale H bond networks. Such behavior is discussed in terms of water activity because the mechanical ECM properties were found to depend on hydration.

Does water activity rule P. mirabilis periodic swarming? II. Viscoelasticity and water balance during swarming.

Following the analysis of the biochemical and functional properties of the P. mirabilis extra cellular matrix performed in the first part of this study, the viscoelasticity of an actively growing colony was investigated in relation to water activity. The results demonstrate that the P. mirabilis colony exhibits a marked viscoelastic character likely due to both cell rafts and exoproduct H-bond networks. Besides, the water loss by evaporation during migration has been measured, whereas the experimental determination of the water diffusion coefficient in agar has allowed us to estimate the net water influx at the agar/colony interface. These data drive us to propose that a periodic increase of the water activity at the colony's periphery, mainly due to the drastic surface to volume ratio increase associated with swarming, causes the periodic and synchronous cessation of migration through the dissociation of exoproduct networks, which in turn strongly alters the matrix viscoelasticity.

pH-Tunable rheological properties of a telechelic cationic polyelectrolyte reversible hydrogel.

Steady shear properties and linear and nonlinear viscoelastic behaviors of a poly(methyl methacrylate)-poly(dimethyl amino ethyl methacrylate)-poly(methyl methacrylate) polymer, (PMMA-PDMAEMA-PMMA), telechelic polymers in salt-free aqueous solution have been investigated as a function of concentration and pH. Above a critical concentration, a transient physical network is formed through an association mechanism between hydrophobic end groups, leading to a gel-like behavior. The gel-like polymer solutions were shown to exhibit a peculiar flow behavior, associated with time fluctuation of the transient first normal stress difference, attributed to orientation effects of the stiff charged polymer chains. The viscoelastic behavior was shown to be governed by two pH dependent time scales: a short time scale controlled by the lifetime of the hydrophobic associative junctions and a long time scale corresponding to the network relaxation time. All rheological results show strong evidence that Coulomb interactions, which control both macromolecular chain rigidity and inter-chain interactions, lead to specific pH-tunable properties of great potential interest.