Abnormally high content of free glucosamine residues identified in a preparation of commercially available porcine intestinal heparan sulfate

Heparan sulfate (HS) polysaccharides are ubiquitous in animal tissues as components of proteoglycans, and they participate in many important biological processes. HS carbohydrate chains are complex and can contain rare structural components such as N-unsubstituted glucosamine (GlcN). Commercially available HS preparations have been invaluable in many types of research activities. In the course of preparing microarrays to include probes derived from HS oligosaccharides, we found an unusually high content of GlcN residue in a recently purchased batch of porcine intestinal mucosal HS. Composition and sequence analysis by mass spectrometry of the oligosaccharides obtained after heparin lyase III digestion of the polysaccharide indicated two and three GlcN in the tetrasaccharide and hexasaccharide fractions, respectively. (1)H NMR of the intact polysaccharide showed that this unusual batch differed strikingly from other HS preparations obtained from bovine kidney and porcine intestine. The very high content of GlcN (30%) and low content of GlcNAc (4.2%) determined by disaccharide composition analysis indicated that N-deacetylation and/or N-desulfation may have taken place. HS is widely used by the scientific community to investigate HS structures and activities. Great care has to be taken in drawing conclusions from investigations of structural features of HS and specificities of HS interaction with proteins when commercial HS is used without further analysis. Pending the availability of a validated commercial HS reference preparation, our data may be useful to members of the scientific community who have used the present preparation in their studies

Unbound states of 32Cl and the 31S(p,\gamma)32Cl reaction rate

The 31S(p,\gamma)32Cl reaction is expected to provide the dominant break-out path from the SiP cycle in novae and is important for understanding enrichments of sulfur observed in some nova ejecta. We studied the 32S(3He,t)32Cl charge-exchange reaction to determine properties of proton-unbound levels in 32Cl that have previously contributed significant uncertainties to the 31S(p,\gamma)32Cl reaction rate. Measured triton magnetic rigidities were used to determine excitation energies in 32Cl. Proton-branching ratios were obtained by detecting decay protons from unbound 32Cl states in coincidence with tritons. An improved 31S(p,\gamma)32Cl reaction rate was calculated including robust statistical and systematic uncertainties

In vivo imaging and quantitative analysis of leukocyte directional migration and polarization in inflamed tissue

Directional migration of transmigrated leukocytes to the site of injury is a central event in the inflammatory response. Here, we present an in vivo chemotaxis assay enabling the visualization and quantitative analysis of subtype-specific directional motility and polarization of leukocytes in their natural 3D microenvironment. Our technique comprises the combination of i) semi-automated in situ microinjection of chemoattractants or bacteria as local chemotactic stimulus, ii) in vivo near-infrared reflected-light oblique transillumination (RLOT) microscopy for the visualization of leukocyte motility and morphology, and iii) in vivo fluorescence microscopy for the visualization of different leukocyte subpopulations or fluorescence-labeled bacteria. Leukocyte motility parameters are quantified off-line in digitized video sequences using computer-assisted single cell tracking. Here, we show that perivenular microinjection of chemoattractants [macrophage inflammatory protein-1alpha (MIP-1alpha/Ccl3), platelet-activating factor (PAF)] or E. coli into the murine cremaster muscle induces target-oriented intravascular adhesion and transmigration as well as polarization and directional interstitial migration of leukocytes towards the locally administered stimuli. Moreover, we describe a crucial role of Rho kinase for the regulation of directional motility and polarization of transmigrated leukocytes in vivo. Finally, combining in vivo RLOT and fluorescence microscopy in Cx3CR1(gfp/gfp) mice (mice exhibiting green fluorescent protein-labeled monocytes), we are able to demonstrate differences in the migratory behavior of monocytes and neutrophils.Taken together, we propose a novel approach for investigating the mechanisms and spatiotemporal dynamics of subtype-specific motility and polarization of leukocytes during their directional interstitial migration in vivo

Spinal infection: state of the art and management algorithm

Spinal infection is a rare pathology although a concerning rising incidence has been observed in recent years. This increase might reflect a progressively more susceptible population but also the availability of increased diagnostic accuracy. Yet, even with improved diagnosis tools and procedures, the delay in diagnosis remains an important issue. This review aims to highlight the importance of a methodological attitude towards accurate and prompt diagnosis using an algorithm to aid on spinal infection management. METHODS: Appropriate literature on spinal infection was selected using databases from the US National Library of Medicine and the National Institutes of Health. RESULTS: Literature reveals that histopathological analysis of infected tissues is a paramount for diagnosis and must be performed routinely. Antibiotic therapy is transversal to both conservative and surgical approaches and must be initiated after etiological diagnosis. Indications for surgical treatment include neurological deficits or sepsis, spine instability and/or deformity, presence of epidural abscess and upon failure of conservative treatment. CONCLUSIONS: A methodological assessment could lead to diagnosis effectiveness of spinal infection. Towards this, we present a management algorithm based on literature findings

PEDOT:PSS-coated polybenzimidazole electroconductive nanofibers for biomedical applications

Bioelectricity drives several processes in the human body. The development of new materials that can deliver electrical stimuli is gaining increasing attention in the field of tissue engineering. In this work, novel, highly electrically conductive nanofibers made of poly [2,20 - m-(phenylene)-5,50 -bibenzimidazole] (PBI) have been manufactured by electrospinning and then coated with cross-linked poly (3,4-ethylenedioxythiophene) doped with poly (styrene sulfonic acid) (PEDOT:PSS) by spin coating or dip coating. These scaffolds have been characterized by scanning electron microscopy (SEM) imaging and attenuated total reflectance Fourier-transform infrared (ATR-FTIR) spectroscopy. The electrical conductivity was measured by the four-probe method at values of 28.3 S·m−1 for spin coated fibers and 147 S·m−1 for dip coated samples, which correspond, respectively, to an increase of about 105 and 106 times in relation to the electrical conductivity of PBI fibers. Human bone marrow-derived mesenchymal stromal cells (hBM-MSCs) cultured on the produced scaffolds for one week showed high viability, typical morphology and proliferative capacity, as demonstrated by calcein fluorescence staining, 40 ,6-diamidino-2-phenylindole (DAPI)/Phalloidin staining and MTT [3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide] assay. Therefore, all fiber samples demonstrated biocompatibility. Overall, our findings highlight the great potential of PEDOT:PSS-coated PBI electrospun scaffolds for a wide variety of biomedical applications, including their use as reliable in vitro models to study pathologies and the development of strategies for the regeneration of electroactive tissues or in the design of new electrodes for in vivo electrical stimulation protocols.info:eu-repo/semantics/publishedVersio

Chondrogenic differentiation of mesenchymal stem/stromal cells on 3D porous poly (ε-caprolactone) scaffolds: Effects of material alkaline treatment and chondroitin sulfate supplementation

Cartilage defects resultant from trauma or degenerative diseases (e.g., osteoarthritis) can potentially be repaired using tissue engineering (TE) strategies combining progenitor cells, biomaterial scaffolds and bio-physical/chemical cues. This work examines promoting chondrogenic differentiation of human bone marrow mesenchymal stem/stromal cells (BM-MSCs) by combining the effects of modified poly (ε-caprolactone) (PCL) scaffolds hydrophilicity and chondroitin sulfate (CS) supplementation in a hypoxic 5% oxygen atmosphere. 3D-extruded PCL scaffolds, characterized by μCT, featured a 21 mm−1 surface area to volume ratio, 390 μm pore size and approximately 100% pore interconnectivity. Scaffold immersion in sodium hydroxide solutions for different periods of time had major effects in scaffold surface morphology, wettability and mechanical properties, but without improvements on cell adhesion. In-situ chondrogenic differentiation of BM-MSC seeded in 3D-extruded PCL scaffolds resulted in higher cell populations and ECM deposition along all scaffold structure, when chondrogenesis was preceded by an expansion phase. Additionally, CS supplementation during BM-MSC expansion was crucial to enhance aggrecan gene expression, known as a hallmark of chondrogenesis. Overall, this study presents an approach to tailor the wettability and mechanical properties of PCL scaffolds and supports the use of CS-supplementation as a biochemical cue in integrated TE strategies for cartilage regeneration

New sulfonated aramide nanoparticles and their copper complexes with anomalous dielectric behavior

ABSTRACT: We report the preparation of thermally stable spherical sulfonated aramides nanoparticles and their copper(II) complexes. Metal chelation with copper ions furnished polymeric complexes in a 1: 2 ratio with square planar geometries as judged by their IR, UV, electron spin resonance, and elemental analysis data. The direct-current electrical conductivities demonstrated the semiconduct-ing nature of the polymeric particles and their copper complexes. Dielectric loss analysis studies showed spectral peaks appearing at characteristic frequencies, which suggested the presence of relaxing dipoles in all of the polymers. All loss peaks were shown on a lin-ear frequency scale and appeared in the range of 1 decade, and no overlap was observed in any of the samples, whereas in the normal polymer’s dielectric loss behavior, each peak covered more than 1 decade. Moreover, the peak positions did not change with increas-ing temperature; this indicated a nonactivated process. The reported dielectric results revealed anomalous behavior, which has not been reported before for such polymeric analogues, as the polarization in these cases was limited by nonthermal forces, and a steady-state constant polarization was produced by an applied field. A simple method for the formation of a microporous semiconducting thin film of a polymer derived from isophthalic acid and diaminodiphenylsulfone is described. VC 2012 Wiley Periodicals, Inc. J. Appl

Sea ice melt pond bathymetry reconstructed from aerial photographs using photogrammetry: a new method applied to MOSAiC data

Melt ponds are a core component of the summer sea ice system in the Arctic, increasing the uptake of solar energy and impacting the ice-associated ecosystem. They were thus one of the key topics during the 1-year drift campaign Multidisciplinary drifting Observatory for the Study of Arctic Climate (MOSAiC) in the Transpolar Drift 2019/2020. Pond depth is a dominating factor in describing the surface meltwater volume; it is necessary to estimate budgets and used in model parameterization to simulate pond coverage evolution. However, observational data on pond depth are spatially and temporally strongly limited to a few in situ measurements. Pond bathymetry, which is pond depth spatially fully resolved, remains unexplored. Here, we present a newly developed method to derive pond bathymetry from aerial images. We determine it from a photogrammetric multi-view reconstruction of the summer ice surface topography. Based on images recorded on dedicated grid flights and facilitated assumptions, we were able to obtain pond depth with a mean deviation of 3.5 cm compared to manual in situ observations. The method is independent of pond color and sky conditions, which is an advantage over recently developed radiometric airborne retrieval methods. It can furthermore be implemented in any typical photogrammetry workflow. We present the retrieval algorithm, including requirements for the data recording and survey planning, and a correction method for refraction at the air–pond interface. In addition, we show how the retrieved surface topography model synergizes with the initial image data to retrieve the water level of individual ponds from the visually determined pond margins. We use the method to give a profound overview of the pond coverage on the MOSAiC floe, on which we found unexpected steady pond coverage and volume. We were able to derive individual pond properties of more than 1600 ponds on the floe, including their size, bathymetry, volume, surface elevation above sea level, and temporal evolution. We present a scaling factor for single in situ depth measurements, discuss the representativeness of in situ pond measurements and the importance of such high-resolution data for new satellite retrievals, and show indications for non-rigid pond bottoms. The study points out the great potential to derive geometric properties of the summer sea ice surface emerging from the increasingly available visual image data recorded from uncrewed aerial vehicles (UAVs) or aircraft, allowing for an integrated understanding and improved formulation of the thermodynamic and hydrological pond system in models.</p

Prehospital tracheal intubations by anaesthetist-staffed critical care teams: a prospective observational multicentre study

Background Prehospital tracheal intubation is a potentially lifesaving intervention, but is associated with prolonged time on-scene. Some services strongly advocate performing the procedure outside of the ambulance or aircraft, while others also perform the procedure inside the vehicle. This study was designed as a non-inferiority trial registering the rate of successful tracheal intubation and incidence of complications performed by a critical care team either inside or outside an ambulance or helicopter. Methods This observational multicentre study was performed between March 2020 and September 2021 and involved 12 anaesthetist-staffed critical care teams providing emergency medical services by helicopter in Denmark, Norway, and Sweden. The primary outcome was first-pass successful tracheal intubations. Results Of the 422 drug-assisted tracheal intubations examined, 240 (57%) took place in the cabin of the ambulance or helicopter. The rate of first-pass success was 89.2% for intubations in-cabin vs 86.3% outside. This difference of 2.9% (confidence interval −2.4% to 8.2%) (two sided 10%, including 0, but not the non-inferiority limit Δ=−4.5) fulfils our criteria for non-inferiority, but not significant superiority. These results withstand after performing a propensity score analysis. The mean on-scene time associated with the helicopter in-cabin procedures (27 min) was significantly shorter than for outside the cabin (32 min, P=0.004). Conclusions Both in-cabin and outside the cabin, prehospital tracheal intubation by anaesthetists was performed with a high success rate. The mean on-scene time was shorter in the in-cabin helicopter cohort.publishedVersio