The interplay between HLA-B27 and ERAP1/ERAP2 aminopeptidases: from anti-viral protection to spondyloarthritis.

The human leukocyte antigen class I gene HLA-B27 is the strongest risk factor for ankylosing spondylitis (AS), a chronic inflammatory arthritic disorder. More recently, the Endoplasmic Reticulum Aminopeptidase (ERAP) 1 and 2 genes have been identified by genome wide association studies (GWAS) as additional susceptibility factors. In the ER, these aminopeptidases trim the peptides to a length suitable to fit into the groove of the major histocompatibility complex (MHC) class I molecules. It is noteworthy that an epistatic interaction between HLA-B27 and ERAP1, but not between HLA-B27 and ERAP2, has been highlighted. However, these observations suggest a paramount centrality for the HLA-B27 peptide repertoire that determines the natural B27 immunological function, i.e. the T cell antigen presentation and, as a by-product, elicits HLA-B27 aberrant behaviours: (i) the misfolding leading to ER stress responses and autophagy and (ii) the surface expression of homodimers acting as ligands for innate immune receptors. In this context, it has been observed that the HLA-B27 carriers, besides being prone to autoimmunity, display a far better surveillance to some viral infections. This review focuses on the ambivalent role of HLA-B27 in autoimmunity and viral protection correlating its functions to the quantitative and qualitative effects of ERAP1 and ERAP2 polymorphisms on their enzymatic activity.

Development of antibacterial and antifungal silver-coated polyurethane foams as air filtration units for the prevention of respiratory diseases.

AIM: The development of silver-coated polyurethane filters as filtration units for the prevention of the respiratory diseases. METHODS AND RESULTS: An innovative silver deposition technology based on the photo-reduction in a silver salt was adopted. The efficacy of the technology in providing a homogeneous distribution of the silver particles was verified by scanning electron microscopy and energy dispersive X-ray spectroscopy. The materials were tested through microbiological procedures used in industry to verify the efficacy of the silver-coated filters on the viability and growth of selected micro-organisms. Direct inoculation test, filtration experiment and shaking tests were performed on microbial human pathogens associated with air filtration units and respiratory disease, namely Legionella pneumophila, Pseudomonas aeruginosa, Aspergillus niger and Aspergillus flavus, by adopting Escherichia coli and Staphylococcus aureus as control organisms. CONCLUSIONS: The results provided evidence of the effectiveness of the silver coating in reducing the bioaerosolization of viable human pathogens into environments using recirculated air. SIGNIFICANCE AND IMPACT OF THE STUDY: Micro-organisms can affect the air quality in indoor environments and can be responsible for infectious, allergic or toxic disturbances on human airways. The development of an adequate bioaerosol control might ameliorate a positive health effect in humans.

Development of silver nano-coatings on silk sutures as a novel approach against surgical infections.

The infections give rise to a range of clinical problems and prolong hospitalization with increased healthcare costs. Moreover, persistent infections exasperate the problem of antibiotic resistance. The aim of this study was the development of effective and low-cost antibacterial silver coatings on surgical sutures by adopting an innovative photochemical deposition process to prevent early contamination of surgical wounds. The silver deposition technology adopted in this work is an innovative process based on the in situ photoreduction of a silver solution. The samples were dipped in the silver solution and then exposed to UV radiation in order to induce the synthesis of silver clusters on the surface of the suture. The homogeneous distribution of silver particles on the surface and on the cross-section of the treated sutures was demonstrated. All the antibacterial studies clearly demonstrated that the use of novel silver treated sutures could represent clinical advantages in terms of the prevention of surgical infections against bacterial colonization. The silver coating deposited on the sutures demonstrated no cytotoxic effect on a selected cell population. The results obtained suggested that the antibacterial silver-coated sutures developed in this work could represent an interesting alternative to conventional sutures, with evident advantages in terms of prevention of the surgical infections and on the health costs. In addiction, very low concentrations of silver significantly inhibited the microbial load, without affecting the cell viability.

Effect of silver nanocoatings on catheters for haemodialysis in terms of cell viability, proliferation, morphology and antibacterial activity.

The onset of infections associated to bacterial proliferation and biofilm formation on indwelling medical devices represents the major risk of morbidity and mortality among patients. In order to contain the risk of infections in clinical practice, there is a growing interest nowadays in silver-based products due to the strong antimicrobial efficacy of silver against a broad spectrum of microorganisms. In this work, temporary catheters for haemodialysis were coated with silver nano-particles through the in situ photo-reduction of a silver salt in alcoholic solution. A homogeneous distribution of silver particles firmly bonded to the substrate was obtained through the adopted technique. An optimisation study was required to define the amount of silver, in order to obtain good efficacy against Gram-positive and Gram-negative bacteria and no cytotoxic effect. At this purpose, three concentrations of silver, 0.1, 0.25 and 0.5 wt%, have been deposited and tested with respect to bacterial reduction percentage and cellular response. Particularly, bacterial enumeration on Escherichia coli and Staphylococcus aureus, and BrdU incorporation, TUNEL assay and Actin staining on a selected primary cell population were performed on catheters treated with the different silver solutions. The silver percentages tested demonstrated strong antibacterial properties together with a good cellular response, thus indicating that the developed product could be proposed in clinical practice and that the lower percentage tested can be preferred with evident advantages in terms of costs.

Genetics of Behçet's disease in Sardinia: two distinct extended HLA haplotypes harbour the B*51 allele in the normal population and in patients.

OBJECTIVES: To define the contribution of HLA genes and extended HLA haplotypes to the susceptibility to Behçet's disease (BD) in Sardinia. METHODS: Forty-five unrelated Sardinian patients with BD, diagnosed according to the ISG criteria, 45 HLA-B*51 positive and 185 unselected healthy controls were enrolled in the study. DNA samples were typed for HLA class I and class II alleles and genotyped for microsatellites (MICA-TM) and single-nucleotide polymorphisms (rs1264457 HLA-E; rs2281820 motilin; rs1799724 at -857, rs361525 at -238 TNF-alpha) spanning the HLA region. RESULTS: HLA-B*5101 was confirmed as conferring susceptibility to BD (pc=0.0042; OR=4.4; 95% CI=2.0 to 9.6). It is noteworthy that in Sardinia this allele was found more frequently within a haplotype (HLA-A2; -Cw2; -B*5101; -DRB1*11; -DQA1*05; - DQB1*03) that reached its highest frequency in patients with BD. Linkage disequilibrium analysis showed the existence of an additional B*51 haplotype (HLA-A2; -Cw2; -B*5101; -DRB1*04; -DQA1*03; -DQB1*03) not associated with susceptibility to the disease. CONCLUSIONS: In Sardinia, the BD-associated HLA-B*5101 allele is inherited as part of two distinctive haplotypes differently distributed in patients and controls. These findings can be interpreted as suggestive of the presence of additional genes within the MHC region conferring susceptibility to BD. The hypothesis that an environmental pressure could have contributed to the preservation of the BD-associated HLA haplotype in Sardinia is also discussed.

Efficacy of silver treated catheters for haemodialysis in preventing bacterial adhesion.

The growing resistance of many strains of bacteria to antibiotics and antiseptics is becoming a serious problem in medicine. Nano-silver is one of the most prominent products in medicine because it exhibits unusual physicochemical properties and a strong biological activity. In this work an innovative silver deposition technology was applied to temporary polyurethane catheters for haemodialysis. The working conditions of catheters were reproduced through laboratory equipment that ensured the flow of deionized water and simulated body fluid inside the lumina at corporeal temperature. The growth and the adhesion of Staphylococcus aureus on the surface of the device were studied through fluorescence microscopy. ICP-AES was adopted to calculate the amount of silver released from the substrate. The stability of the coating during the whole working life of the device was demonstrated through thermo-gravimetric analysis.

Antibacterial coatings on haemodialysis catheters by photochemical deposition of silver nanoparticles.

Antibacterial coatings on catheters for acute dialysis were obtained by an innovative and patented silver deposition technique based on the photo-reduction of the silver solution on the surface of catheter, with consequent formation of antibacterial silver nanoparticles. Aim of this work is the structural and morphological characterization of these medical devices in order to analyze the distribution and the size of clusters on the polymeric surface, and to verify the antibacterial capability of the devices treated by this technique against bacterial proliferation. The structure and morphology of the silver nanoparticles were investigated by using scanning and transmission electron microscopy. The antimicrobial capability of the catheters after silver deposition was confirmed by antibacterial tests with Escherichia coli. Both scanning electron microscopy analysis and antibacterial tests were performed also after washing catheters for 30 days in deionized water at 37°C, relating these data to thermogravimetric analysis and to energy dispersive spectroscopy, in order to check the resistance of coating and its antimicrobial capability after the maximum time of life of these devices.

A functional polymorphism of the vasoactive intestinal peptide receptor 1 gene correlates with the presence of HLA-B*2705 in Sardinia.

The association of HLA-B27 with ankylosing spondylitis (AS) is the strongest among all inflammatory diseases. However, the exact role of these molecules in disease pathogenesis is still unknown. The existence of HLA-B27 variants rarely found in patients introduces a further level of complexity. It is now accepted that other genes of minor impact contribute to modify disease susceptibility and these genes might be diverse in different populations depending on the genetic background. We report here a study performed in Sardinia, an outlier population in which two major HLA-B27 subtypes are present, B (*)2705 strongly associated with AS and B (*)2709 which is not, and show the co-occurrence of the B (*)2705 allele with a single nucleotide polymorphism (SNP) mapping at 3'-UTR of the receptor 1 (VIPR1) for the vasoactive intestinal peptide (VIP), a neuropeptide with anti-inflammatory properties. This same SNP is associated with a different kinetics of down-modulation of the VIPR1 mRNA in monocytes after exposure to lipopolysaccharide (P=0.004). This particular setting, HLA-B (*)2705 and a functional polymorphism in VIPR1 gene, might be due to a founder effect or might be the result of a selective pressure. Irrespectively, the consequent downregulation of this receptor in the presence of a 'danger' signal might influence susceptibility to AS.

Surface chemical and biological characterization of flax fabrics modified with silver nanoparticles for biomedical applications.

Silver nanophases are increasingly used as effective antibacterial agent for biomedical applications and wound healing. This work aims to investigate the surface chemical composition and biological properties of silver nanoparticle-modified flax substrates. Silver coatings were deposited on textiles through the in situ photo-reduction of a silver solution, by means of a large-scale apparatus. The silver-coated materials were characterized through X-ray Photoelectron Spectroscopy (XPS), to assess the surface elemental composition of the coatings, and the chemical speciation of both the substrate and the antibacterial nanophases. A detailed investigation of XPS high resolution regions outlined that silver is mainly present on nanophases' surface as Ag2O. Scanning electron microscopy and energy dispersive X-ray spectroscopy were also carried out, in order to visualize the distribution of silver particles on the fibers. The materials were also characterized from a biological point of view in terms of antibacterial capability and cytotoxicity. Agar diffusion tests and bacterial enumeration tests were performed on Gram positive and Gram negative bacteria, namely Staphylococcus aureus and Escherichia coli. In vitro cytotoxicity tests were performed through the extract method on murine fibroblasts in order to verify if the presence of the silver coating affected the cellular viability and proliferation. Durability of the coating was also assessed, thus confirming the successful scaling up of the process, which will be therefore available for large-scale production.

Asymmetrical dripping.

Dripping from a faucet is studied, where by cutting diagonally the tip breaks the cylindrical symmetry. We measured long dripping time series recorded with a laser-beam apparatus and continuous change in drop position using a high-speed camera. It is found that the added degree of freedom produces a transversal oscillation of a pending drop, which couples with a vertical oscillation induced by the breaking off of the previous drop. As a consequence dripping times shorten and dripping patterns regularize. The effect is attributed to the "reduced" contact circle and to decreased frequency of the vertical oscillations of the residue. A very complex flow circulation pattern within the forming drop is observed. It is suggested that geometrical shape vibrations of the pending drop take place by the development of eddies of different amount, more effective in the vicinity of the highest edge of the tip, where they dissipate more slowly. This asymmetrical liquid flow is brought about by the slanting form of the orifice and overlaps with the axial oscillations.

Experimental study of dripping dynamics.

The dependence of dripping dynamics from physical properties of the nozzles is investigated. The analysis is performed by means of two complementary methods: (i) long dripping time series recorded with a standard laser-beam apparatus; and (ii) drop formations observed with a fast digital camcorder. Dripping from nozzles of different sizes is analyzed, and the formation of satellite drops is related to the preeminent physical parameter of control (flow rate). Quasielastic collisions between parent and satellite drops are observed.

[Our experience in treating femoral neck fractures. Comments on cases treated from 1972 to 1984. A clinico-statistical study]. / La nostra esperienza sul trattamento delle fratture del collo di femore. Considerazioni sui casi trattati dal 1972 al 1984. Indagine clinico-statistica.

The Authors report the clinico-statistic results concerning 550 cases of femur neck fractures, representing the casuistry of 12 years' activity developed by the Ortopedico-Traumatological Department of the Hospital Institutes of the Association of the Bergamo Lowland Communes, USSL nr. 32 (from 1972 up to 1984). After considering the main etiologic factors of this injury, the Authors study the surgical techniques adopted, discuss the indications, state the results obtained, and do some remarks about the patients' functional and social recovery. The Authors emphasize the importance of an early and valid osteosynthesis allowing a quicker functional recovery, a reduction in the times of stay in bed and an extension in the average survival of such patients.

In vivo testing of silver treated fibers for the evaluation of skin irritation effect and hypoallergenicity.

Textiles are a fertile breeding ground for a multitude of micro-organisms under appropriate conditions of moisture and temperature. The broad-spectrum biocide properties of silver are well known and many technologies have been developed so far to treat textiles with silver. The efficacy of the silver deposition technology presented in this article has been already demonstrated in previous works, where the strong adhesion of silver nanoparticles to the substrate and their antibacterial capability have been assessed. This work focuses on the evaluation of any possible interaction of silver treated cotton with human skin, in terms of skin irritation and hypoallergenicity. Moreover, the presence of silver and the antibacterial capability against Gram positive and Gram negative bacteria, namely Staphylococcus aureus and Escherichia coli, were verified even after several washing cycles in order to develop a product with long-term antibacterial capability and no adverse effects in terms of skin irritation and hypoallergenicity.

Age-dependent association of idiopathic achalasia with vasoactive intestinal peptide receptor 1 gene.

Idiopathic achalasia is a rare disorder of the oesophagus of unknown aetio-pathogenesis characterized by a myenteric inflammation, aperistalsis and insufficient lower oesophageal sphincter relaxation. Vasoactive intestinal peptide (VIP), present in the myenteric plexus, is involved in smooth muscle relaxation and acts as an anti-inflammatory cytokine. The human VIP receptor 1 gene (VIPR1) is highly polymorphic and may play a role in idiopathic achalasia. One hundred and four consecutive patients and 300 random controls from the same geographic area were typed for five SNPs mapping in the VIPR1 gene. Patients with idiopathic achalasia show a significant difference in allele, genotype and phenotype distribution of SNP rs437876 mapping in intron 4. This association, however, was almost entirely due to the group of patients with late disease onset (P = 0.0005). These results strongly suggest that idiopathic achalasia is a heterogeneous disease with a different aetiology in cases with early or late disease onset.