The use of physical hydrogels of chitosan for skin regeneration following third-degree burns

Skin repair is an important field of the tissue engineering, especially in the case of extended third-degree burns, where the current treatments are still insufficient in promoting satisfying skin regeneration. Bio-inspired bi-layered physical hydrogels only constituted of chitosan and water were processed and applied to the treatment of full-thickness burn injuries. The aim of the study was at assessing whether this material was totally accepted by the host organism and allowed in vivo skin reconstruction of limited area third-degree burns. A first layer constituted of a rigid protective gel ensured good mechanical properties and gas exchanges. A second soft and flexible layer allowed the material to follow the geometry of the wound and ensured a good superficial contact. To compare, highly viscous solutions of chitosan were also considered. Veterinary experiments were performed on pig's skins and biopsies at days 9, 17, 22, 100 and 293, were analysed by histology and immuno-histochemistry. Only one chitosan material was used for each time. All the results showed that chitosan materials were well tolerated and promoted a good tissue regeneration. They induced inflammatory cells migration and angiogenetic activity favouring a high vascularisation of the neo-tissue. At day 22, type I and IV collagens were synthesised under the granulation tissue and the formation of the dermal-epidermal junction was observed. After 100 days, the new tissue was quite similar to a native skin, especially by its aesthetic aspect and its great flexibility.     

Molecular characterization of measles virus circulating in the Indian Ocean Islands during 2005-2006 and in France in 2006

Despite the availability of safe and immunogenic vaccines, measles still causes significant morbidity and mortality especially in Africa. In this study, two measles outbreaks in the Indian Ocean Islands; Mayotte in 2005-2006 and Seychelles in 2006 were studied. Nasopharyngeal swabs, urine and/or blood samples were collected from patients with clinically diagnosed measles. Measles viruses were isolated in four cases from patients in Mayotte. Measles strains circulating in both outbreaks were determined to be genotype D4 when compared to the WHO reference strains. During this time, measles virus was isolated from patients in France and they were also found to belong to the same genotype. The viruses clustered into two distinct D4 subgroups; The Indian Ocean strains were similar to the Montreal-subgroup, whereas the French strains associated with the Johannesburg-subgroup. The Indian Ocean strains formed a homogeneous group. They shared four specific amino acids in the 3' region of the N gene and two amino acids in the H gene, which differed from other genotype D4 viruses. This suggests that the same measles lineage circulated in Mayotte and Seychelles. Sequence comparison of the French isolates with other measles strains showed that they were more closely related to strains circulating in Germany in 2005, which had their origin in Romania. This study provides the baseline for molecular epidemiology of measles virus in Mayotte and Seychelles. The knowledge of circulating measles virus will help in documenting measles elimination program. This report also highlights the fact that progress of measles elimination is blighted continually by the phenomenon of measles importation.     

Innate immunity, assessed by plasma NO measurements, is not suppressed during the incubation fast in eiders

Immunity is hypothesized to share limited resources with other physiological functions and may mediate life history trade-offs, for example between reproduction and survival. However, vertebrate immune defense is a complex system that consists of three components. To date, no study has assessed all of these components for the same animal model and within a given situation. Previous studies have determined that the acquired immunity of common eiders (Somateria mollissima) is suppressed during incubation. The present paper aims to assess the innate immune response in fasting eiders in relation to their initial body condition. Innate immunity was assessed by measuring plasma nitric oxide (NO) levels, prior to and after injection of lipopolysaccharides (LPS), a method which is easily applicable to many wild animals. Body condition index and corticosterone levels were subsequently determined as indicators of body condition and stress level prior to LPS injection. The innate immune response in eiders did not vary significantly throughout the incubation period. The innate immune response of eiders did not vary significantly in relation to their initial body condition but decreased significantly when corticosterone levels increased. However, NO levels after LPS injection were significantly and positively related to initial body condition, while there was a significant negative relationship with plasma corticosterone levels. Our study suggests that female eiders preserve an effective innate immune response during incubation and this response might be partially determined by the initial body condition.     

Calcium-deficient apatite synthesized by ammonia hydrolysis of dicalcium phosphate dihydrate: influence of temperature, time, and pressure

In this work, calcium-deficient apatites (CDA) were synthesized by ammonia hydrolysis reaction of dicalcium phosphate dihydrate (DCPD; CaHPO4 x 2 H2O) to obtain biphasic calcium phosphates (BCP) without any extraionic substitution. The influence of three parameters was studied: temperature of the reaction (70 and 100 degrees C), time of the reaction (4 and 18 h), and the pressure (open and closed system). Experiments were made according to a factorial design method (FDM) allowing optimization of the number of samples as well as statistical analysis of results. Moreover, the influence of temperature (until 200 degrees C) was investigated. The crystal size of CDA was determined according to the Scherrer's formula and from Rietveld refinements taking the CDA anisotropy into account. The last method seems to be a reliable method to determine crystallite sizes of CDA, since crystallite sizes of CDA along <00l> and directions were accessible. The results describe the hydroxyapatite % (HA%) in BCP by a first-order polynomial equation in the experimental area studied and the HA content was found to increase by raising time and temperature of the reaction. Moreover, the type of reaction system (open/closed vessel) appeared to have little influence on HA%.     

Retinal degenerations: from cell signaling to cell therapy; pre-clinical and clinical issues

Extracellular signaling molecules have been implicated in the progression of Retinal Degeneration (RD). Gene regulatory events linked to the maintenance of retinal structure and function incorporate signaling cascades that may serve as therapeutic targets for some forms of blindness. This review shall focus on the evidence for non-cell-autonomous mechanisms that affect the pattern of degeneration seen in retinal dystrophies, the types of signals that may influence the course of degeneration and finally with the related prospects for retinal-therapies.     

Eubacterial diversity of activated biomass from a common effluent treatment plant

A common effluent treatment plant (CETP) is a biological wastewater treatment facility that receives wastewater from different industries. The activated biomass in the CETP survives on a wide range of chemicals with no fixed wastewater characteristics. We carried out a diversity analysis of this activated biomass using culture as well as culture-independent techniques. Using culture-based techniques, strains belonging to 26 different genera from the phyla Proteobacteria, Actinobacteria and Firmicutes were isolated. The gamma-proteobacteria was the best represented class, with 36.5% of the isolates. Bacterial diversity was also analyzed culture-independently by means of sequence determination of cloned 16S rRNA genes. Twenty-one different genera from the phyla Proteobacteria, Firmicutes, Planctomycetes and Bacteroidetes were identified. The total diversity of the activated biomass was composed of members of five known phyla, represented by 37 genera, with the Proteobacteria constituting the most abundant phylum detected. However, a very large fraction of the diversity represented a hitherto unidentified bacterial population. More than half (50.2%) of the 16S rDNA clones represented unidentified non-culturable bacteria, underlining the vast untapped diversity of CETP communities. Our results also indicate that both culture-based and culture-independent techniques should be combined to cover the microbial diversity of complex ecosystems.     

Influenza A virus elevates active cathepsin B in primary murine DC

Dendritic cells (DCs) act as a first-line recognition system for invading pathogens, such as influenza A. The interaction of DC with influenza A virus results in DC activation via endosomal Toll-like receptors and also leads to presentation of viral peptides on MHC class II molecules. Prior work demonstrated that influenza A virus (A/HKx31; H3N2) infection of BALB/c mice activates lung DCs for antigen presentation, and that the enhanced function of these cells persists long after viral clearance and resolution of the virus-induced inflammatory response. Whether influenza A virus has acute or longer-lasting effects on the endo/lysosomal antigen-processing machinery of DCs has not been studied. Here, we show that antigen presentation from intact protein antigen, but not peptide presentation, results in increased T cell stimulation by influenza-exposed lung DCs, suggesting increased antigen processing/loading in these DCs. We find that cathepsin (Cat) B levels and activity are substantially up-regulated in murine lung DCs, harvested 30 days after A/HKx31 infection. CatB levels and activity are also increased in murine splenic and bone marrow-derived DCs, following short-term in vitro exposure to UV-inactivated influenza A virus. Modest effects on CatX are also seen during in vivo and in vitro exposure to influenza A virus. Using a cell permeable Cat inhibitor, we show Cats in influenza-exposed DCs to be functional and required for generation of a T cell epitope from intact ovalbumin. Our findings indicate that influenza A virus affects the MHC class II antigen-processing pathway, an essential pathway for CD4(+) T cell activation.     

Classic scrapie in sheep with the ARR/ARR prion genotype in Germany and France

In the past, natural scrapie and bovine spongiform encephalopathy (BSE) infections have essentially not been diagnosed in sheep homozygous for the A136R154R171 haplotype of the prion protein. This genotype was therefore assumed to confer resistance to BSE and classic scrapie under natural exposure conditions. Hence, to exclude prions from the human food chain, massive breeding efforts have been undertaken in the European Union to amplify this gene. We report the identification of 2 natural scrapie cases in ARR/ARR sheep that have biochemical and transmission characteristics similar to cases of classic scrapie, although the abnormally folded prion protein (PrP(Sc)) was associated with a lower proteinase-K resistance. PrP(Sc) was clearly distinct from BSE prions passaged in sheep and from atypical scrapie prions. These findings strongly support the idea that scrapie prions are a mosaic of agents, which harbor different biologic properties, rather than a unique entity.     

Phenotypic similarity of transmissible mink encephalopathy in cattle and L-type bovine spongiform encephalopathy in a mouse model

Transmissible mink encepholapathy (TME) is a foodborne transmissible spongiform encephalopathy (TSE) of ranch-raised mink; infection with a ruminant TSE has been proposed as the cause, but the precise origin of TME is unknown. To compare the phenotypes of each TSE, bovine-passaged TME isolate and 3 distinct natural bovine spongiform encephalopathy (BSE) agents (typical BSE, H-type BSE, and L-type BSE) were inoculated into an ovine transgenic mouse line (TgOvPrP4). Transgenic mice were susceptible to infection with bovine-passaged TME, typical BSE, and L-type BSE but not to H-type BSE. Based on survival periods, brain lesions profiles, disease-associated prion protein brain distribution, and biochemical properties of protease-resistant prion protein, typical BSE had a distint phenotype in ovine transgenic mice compared to L-type BSE and bovine TME. The similar phenotypic properties of L-type BSE and bovine TME in TgOvPrP4 mice suggest that L-type BSE is a much more likely candidate for the origin of TME than is typical BSE.