In-vitro viability of laser cladded Fe-based metallic glass as a promising bioactive material for improved osseointegration of orthopedic implants.

The commonly used metallic biomaterials fail to prove durability for orthopedics due to their lack of biocompatibility and poor bioactivity which weakens the bonding to bones. Metallic glasses (MGs) have attracted attention as an alternative biomaterial for orthopedics owing to their superior mechanical properties and acceptable biocompatibility. Nevertheless, their uses are limited due to geometrical constraints and brittleness. In this research, the in-vitro bioactivity of laser cladded FeCrMoCB MG on nickel-free stainless-steel was investigated. The proposed MG coating exhibited a remarkable in-vitro bioactivity behavior without prior treatment after immersion in simulated body fluid which is a key factor for better osseointegration. The surface morphology showed that apatite nucleated from the first day and completely covered the surface after 21 days. The energy dispersive spectroscopy spectra showed an increase in the Ca/P ratio from 0.51 at 3 days to 1.61 at 21 days, thus approaching the stoichiometric ratio of bone apatite. The infra-red examination revealed the existence of Ca+2, PO4-2 and OH- indicating the nucleation of brushite and B-type apatite. Additionally, the X-ray diffraction examination revealed the existence of amorphous and nanocrystalline calcium phosphates. These results show the potential of FeCrMoCB MGs as a promising bioactive coating for excellent osseointegration of metallic implants with bone tissue.

Skin collagen can be accurately quantified through noninvasive optical method: Validation on a swine study.

BACKGROUND/PURPOSE: Diffuse reflectance spectroscopy (DRS) is a noninvasive optical technology characterized by relatively low system cost and high efficiency. In our previous study, we quantified the relative concentration of collagen for the individual keloid patient. However, no actual value of collagen concentration can prove the reliability of collagen detection by our DRS system. METHODS: Skin-mimicking phantoms were prepared using different collagen and coffee concentrations, and their chromophore concentrations were quantified using the DRS system to analyze the influence of collagen and other chromophores. Moreover, we used the animal study to compare the DRS system with the collagen evaluation of biopsy section by second-harmonic generation (SHG) microscopy at four different skin parts. RESULTS: In the phantom study, the result showed that coffee chromophore did not severely interfere with collagen concentration recovery. In the animal study, a positive correlation (r=.902) between the DRS system and collagen evaluation with SHG microscopy was found. CONCLUSIONS: We have demonstrated that the DRS system can quantify the actual values of collagen concentration and excluded the interference of other chromophores in skin-mimicking phantoms. Furthermore, a high positive correlation was found in the animal study with SHG microscopy. We consider that the DRS is a potential technique and can evaluate skin condition objectively.

Affinity maturation of T-cell receptor-like antibodies for Wilms tumor 1 peptide greatly enhances therapeutic potential.

WT1126 (RMFPNAPYL) is a human leukocyte antigen-A2 (HLA-A2)-restricted peptide derived from Wilms tumor protein 1 (WT1), which is widely expressed in a broad spectrum of leukemias, lymphomas and solid tumors. A novel T-cell-receptor (TCR)-like single-chain variable fragment (scFv) antibody specific for the T-cell epitope consisting of the WT1/HLA-A2 complex was isolated from a human scFv phage library. This scFv was affinity-matured by mutagenesis combined with yeast display and structurally analyzed using a homology model. This monovalent scFv showed a 100-fold affinity improvement (dissociation constant (KD)=3 nm) and exquisite specificity towards its targeted epitope or HLA-A2(+)/WT1(+) tumor cells. Bivalent scFv-huIgG1-Fc fusion protein demonstrated an even higher avidity (KD=2 pm) binding to the T-cell epitope and to tumor targets and was capable of mediating antibody-dependent cell-mediated cytotoxicity or tumor lysis by chimeric antigen receptor-expressing human T- or NK-92-MI-transfected cells. This antibody demonstrated specific and potent cytotoxicity in vivo towards WT1-positive leukemia xenograft that was HLA-A2 restricted. In summary, T-cell epitopes can provide novel targets for antibody-based therapeutics. By combining phage and yeast displays and scFv-Fc fusion platforms, a strategy for developing high-affinity TCR-like antibodies could be rapidly explored for potential clinical development.

Photobacterium damselae subsp. piscicida responds to antimicrobial peptides through phage-shock-protein A (PspA)-related extracytoplasmic stress response system.

AIMS: To investigate whether Photobacterium damselae subsp. piscicida (Phdp) can sense and directly respond to the presence of cationic antimicrobial peptides (AMPs). METHODS AND RESULTS: We performed proteomic methodologies to investigate the responsive proteins of Phdp on exposure to AMP Q6. Proteins significantly altered were analysed by two-dimensional gel electrophoresis (2-DE) and LC-ESI-Q-TOF MS/MS, thus resulting in five outer membrane proteins (OMPs), seven inner membrane proteins (IMPs) and 17 cytoplasmic proteins (CPs) identified. Quantitative real-time PCR was also applied to monitor the mRNA expression level of these target proteins. CONCLUSIONS: COG analysis revealed that upon exposure to AMP Q6, the majority of the upregulated proteins were involved in signal transduction mechanism, carbohydrate transport and metabolism, post-translational modification, protein turnover and chaperones, while the downregulated proteins were mainly related to energy production and conversion. Among them, phage-shock-protein A (PspA)-related stress response system was considered to play a crucial role. SIGNIFICANCE AND IMPACT OF THE STUDY: To the best of our knowledge, this is the first report elucidating Phdp AMP-response mechanism using proteomics approach. AMP-responsive proteins identified in this study could serve as attractive targets for developing more effective antimicrobial agents against Phdp and other marine bacterial pathogens.

Electrical and optical properties of NiO composite films by radio frequency magnetron sputtering.

The 100-nm NiO-Ag composite films with Ag content of 0 to 24.4 at.% are deposited on glass substrates. It is found that an ultra high electric resistivity (rho) value is obtained and cannot be detected by four point probe measurement when the Ag content is less than 3.4 at.%. The rho value is reduced significantly to 29.0 Omega-cm as Ag content is increased to 4.2 at.%, and it decreases greatly to 0.009 Omega-cm as the content of Ag is further increased to 24.4 at.%. The NiO-Ag composite film with Ag content of 4.2 at.% shows p-type conduction. However, it becomes n-type when the Ag content increases to 9.3 at.%, which results from the Ag atoms segregated at grain boundary of NiO when the excess Ag atoms are added into NiO films. On the other hand, the transmittance of the NiO-Ag films drops continuously from 96.3% to 31.6% as the Ag content increases from 0 to 24.4%.

Proteomic analysis reveals responsive proteins of Vibrio parahaemolyticus on exposure to cationic antimicrobial peptides.

AIMS: To investigate whether Vibrio parahaemolyticus can sense and directly respond to the presence of cationic antimicrobial peptides (AMPs). METHODS AND RESULTS: We performed proteomic methodologies to investigate the responsive proteins of V. parahaemolyticus on exposure to AMP Q6. Differential expression patterns of the outer membrane, inner membrane and cytoplasmic proteins (OMPs, IMPs and CPs) from the bacteria with or without Q6 treatment were obtained using two-dimensional gel electrophoresis (2-DE). Three OMPs (maltoporin, flagellin and OmpV), two IMPs (ATP synthase F1, alpha subunit; and OmpV) and three CPs (pyruvate dehydrogenase subunit E1, glyceraldehyde-3-phosphate dehydrogenase and inositol-5-monophosphate dehydrogenase) were identified using LC-ESI-Q-TOF MS/MS and Mascot program. Real-time quantitative polymerase chain reaction was also performed to determine the mRNA expression level of the target genes. CONCLUSIONS: Our results suggested that V. parahaemolyticus may directly respond to AMPs through the upregulation of the efflux channel, increased yield of energy, effective repair of damaged membranes and downregulation of carbohydrate and nucleotide metabolism for energy preservation. SIGNIFICANCE AND IMPACT OF THE STUDY: AMP-responsive proteins identified in the current study could serve as attractive targets for developing more effective antimicrobial agents.

Proteomic identification of membrane proteins regulating antimicrobial peptide resistance in Vibrio parahaemolyticus.

AIMS: To identify proteins regulating antimicrobial peptide (AMP) resistance in Vibrio parahaemolyticus using membrane subproteome analysis. METHODS AND RESULTS: Three synthetic AMPs (Q4, Q6 and H1) and a natural one from fish (pleurocidin) were used for selection of AMP-resistant strains. Differential expression patterns of the outer and inner membrane proteins (OMPs and IMPs) among wild-type and the resistant strains were obtained using two-dimensional gel electrophoresis. Two OMPs (TolC and flagellin) and five IMPs [transcription termination factor (NusA), long-chain fatty acid transport protein (FadL), elongation factor Tu (EF-Tu), ATP synthase F1, alpha subunit (F1-ATPa) and dihydrolipoamide dehydrogenase (DLD)] were identified using LC-ESI-Q-TOF MS/MS and Mascot program. Real-time quantitative polymerase chain reaction was also performed to determine the mRNA expression level of the target genes. All seven membrane proteins except FadL were upregulated in the AMP-resistant clones, both in the translational and transcriptional levels. CONCLUSIONS: Our results suggested that V. parahaemolyticus may obtain their resistance against AMPs through upregulation of the multidrug efflux transporter, effective repair of damaged membranes and prevention of cellular penetration of AMPs. SIGNIFICANCE AND IMPACT OF THE STUDY: To the best of our knowledge, this is the first report describing bacterial AMP resistance mechanism using proteomic methodologies. Elucidating the mechanism could help in the development of more sustainable antimicrobial agents.

Combined immersion and oral vaccination of Vietnamese catfish (Pangasianodon hypophthalmus) confers protection against mortality caused by Edwardsiella ictaluri.

Edwardsiella ictaluri septicemia occurs worldwide and causes high mortality and considerable economic damage to the catfish industry especially in Vietnam and the USA. To control Edwardsiella septicemia farmers extensively use antibiotics and various vaccination methods. Vaccination with inactivated vaccines has come with variable efficacy. In this trial the results of an approach of controlling Edwardsiella septicemia of Tra catfish (Pangasianodon hypophthalmus) in Vietnam through vaccination via mucosal surfaces are presented. The results show that a combination of primary vaccination by immersion with inactivated E. ictaluri followed by an oral boost with a formulated antigen preparation induces a statistically significant level of protection against mortality caused by experimental infection 4 weeks post-boost. Fish immunized by immersion only show significantly lower level of protection but significantly higher than the controls. Repeated boosts result in improved duration of immunity with a relative percent survival (RPS) of 47% at 90% control mortality. The immunization procedure provides an alternative for disease control through vaccination.

Salt-responsive outer membrane proteins of Vibrio anguillarum serotype O1 as revealed by comparative proteome analysis.

AIMS: Vibrio anguillarum is a universal marine pathogen causing vibriosis. Vibrio anguillarum encounters different osmolarity conditions between seawater and hosts, and its outer membrane proteins (OMPs) play a crucial role in the adaptation to changes of the surroundings. In the present study, proteomic approaches were applied to investigate the salt-responsive OMPs of V. anguillarum. METHODS AND RESULTS: Lower salinity (0.85% NaCl) is more suitable for growth, survival and swimming motility of the bacterium. Comparative two-dimensional electrophoresis (2-DE) analysis reveals six differentially expressed protein spots among three different salinities, which were successfully identified as OmpU, maltoporin, flagellin B, Omp26La, Omp26La and OmpW respectively. CONCLUSIONS: OmpW and OmpU were highly expressed at 3.5% salinity, suggesting their role in the efficient efflux of NaCl. Maltoporin was downregulated in higher salinity, indicating that higher osmolarity inhibits carbohydrate transport and bacterial growth. Omp26La, the homologue of OmpV, functions as a salt-responsive protein in lower salinity. SIGNIFICANCE AND IMPACT OF THE STUDY: To the best of our knowledge, this is the first report describing salt stress-responsive proteins of V. anguillarum using proteomic approaches. Our results provide a useful strategy for delineating the osmoregulatory mechanism of the marine pathogens.

Comparison of human, primate, and canine femora: implications for biomaterials testing in total hip replacement.

The canine model remains an animal of choice for determining the efficacy and safety of various materials and designs used in human total hip replacement (THR). The primate also is used in orthopedic-related research for studying limb anatomy, gait, and age-related bone loss. In order to better understand the appropriateness of these animal models for human THR, external morphologies of thirty-three adult Caucasian human, sixteen adult chimpanzee, and forty-two adult greyhound femora were compared using osteometric methods. Measured parameters included anteversion angle, cervico-diaphyseal angle, femoral head offset in the frontal plane, and anterior bow profiles along the femoral diaphysis. Although some of the measured parameters were approximately similar between species (e.g., mean cervico-diaphyseal angle of humans and chimpanzees), the majority demonstrated morphologic differences that may be biomechanically significant for interpreting stress transfer across the hip (e.g., mean anteversion angle and mean normalized femoral head offset between species). Additionally, age-related changes in proximal femoral morphology and gait pattern, as well as species-related differences in local muscle and inertial forces, may result in notably different loading conditions across the hip joint of each species. Therefore, discretion must be exercised when evaluating canine or primate THR materials and designs for potential use in the human hip.

p(42)Be neutron therapy beams: dose rate and penetration as a function of target thickness and beam filtration.

It is shown that, in the production of p(42)Be neutron beams for clinical use, the use of semithick targets leads to more desirable beam characteristics when appropriate backstop materials are used. Furthermore, an algebraic representation of beam penetration and of dose per unit charge on target, including hardening by polyethylene filters, provides a method for target optimization.

The influence of target thickness and backstop material on proton-produced neutron beams for radiotherapy.

Results are presented of measurements of skin sparing, penetration and total dose per unit of incident charge for various target thicknesses and filtrations for a neutron beam generated by 42 MeV protons on beryllium. These results are contrasted to predictions outlined in a previous paper. The differences from these predictions are attributed to the contribution of low-energy neutrons produced by the residual proton beam in the thick copper target backstop.

Dosimetric measurements in the neutron field produced by a deuterium gas target with a compact medical cyclotron.

Several dosimetric measurements have been carried out in the neutron field produced by accelerated deuterons incident upon a pressurized deuterium gas target. A significant improvement has been achieved in central-axis depth dose when compared to the 9Be(3He,n)11C interaction previously studied . Designs for a target with a high-power dissipation capability are currently underway.