The Continuing Challenges of Translational Research: Clinician-Scientists’ Perspective

Over the last twenty years, revolutionary advances in biomedicine including gene therapy, stem cell research, proteomics, genomics and nanotechnology have highlighted the progressive need to restructure traditional approaches to basic and clinical research in order to facilitate the rapid, efficient integration and translation of these new technologies into novel effective therapeutics. Over the past ten years, funding bodies in the USA and UK such as the National Institutes of Health (NIH) and the Medical Research Council (MRC) have been driving translational research by defining and tackling the hurdles but more still remains to be achieved. This article discusses the ongoing challenges translational researchers face and outlines recent initiatives to tackle these including the new changes to translational funding schemes proposed by the NIH and the MRC and the launch of the "European Advanced Translational Research InfraStructure in Medicine" (EATRIS). It is anticipated that initiatives such as these will not only strengthen translational biomedical research programmes already initiated but should lead to rapid benefits to patients and society

Prevalence of Campylobacter spp. in diarrhoea samples from patients in New South Wales, Australia

Campylobacteriosis is a leading cause of bacterial foodborne disease in many industrialized countries including Australia. New South Wales (NSW) is the most populous state in Australia yet the lack of any Campylobacter species surveillance programs has led to a knowledge gap in the importance of these pathogens as causes of diarrhoea. The data collected in this study demonstrated a need for such programs. In this study, 400 human clinical fecal samples were collected from two NSW locations, Western Sydney and Wagga Wagga, and tested for the presence of Campylobacter spp. Patients were clustered by location, age and gender to assess Campylobacter spp. prevalence within these groups between the two regions. The frequency of Campylobacter spp. was higher in males compared to females in the age groups 0–4 and 5–14 years; 6.4% and 1.0%, and 8.2% and none, respectively (P < 0.05). A second peak was noted in elderly adults compared with those in younger age groups. Based on the findings of the quantitative PCR analysis it was estimated that the age-adjusted prevalence of Campylobacter spp. associated diarrhoea was 159 cases per 100,000 persons. [Int Microbiol 2016; 19(1):33-37]Keywords: Campylobacter species · campylobacteriosis · foodborne diseases · prevalence of pathogens · New South Wales, Australi

Polymer–magnetic composite fibers for remote-controlled drug release

An efficient method is reported, for the fabrication of composite microfibers that can be magnetically actuated and are biocompatible, targeting controlled drug release. Aqueous solutions of polyvinyl alcohol, incorporated with citric acid-coated Fe₃O₄ magnetic nanoparticles (MNPs), are subject to infusion gyration to generate 100–300 nm diameter composite fibers, with controllable MNP loading. The fibers are stable in polar solvents, such as ethanol, and do not show any leaching of MNPs for over 4 weeks. Using acetaminophen as an example, we demonstrate that this material is effective in immobilization and triggered release of drugs, which is achieved by a moving external magnetic field. The remote actuation ability, coupled with biocompatibility and lightweight property, renders enormous potential for these fibers to be used as a smart drug release agent

An observational prospective study of topical acidified nitrite for killing methicillin-resistant Staphylococcus aureus (MRSA) in contaminated wounds

Background Endogenous nitric oxide (NO) kills bacteria and other organisms as part of the innate immune response. When nitrite is exposed to low pH, NO is generated and has been used as an NO delivery system to treat skin infections. We demonstrated eradication of MRSA carriage from wounds using a topical formulation of citric acid (4.5%) and sodium nitrite (3%) creams co-applied for 5 days to 15 wounds in an observational prospective pilot study of 8 patients. Findings Following treatment with topical citric acid and sodium nitrite, 9 of 15 wounds (60%) and 3 of 8 patients (37%) were cleared of infection. MRSA isolates from these patients were all sensitive to acidified nitrite in vitro compared to methicillin-sensitive S. aureus and a reference strain of MRSA. Conclusions Nitric oxide and acidified nitrite offer a novel therapy for control of MRSA in wounds. Wounds that were not cleared of infection may have been re-contaminated or the bioavailability of acidified nitrite impaired by local factors in the tissue

Development and characterisation of a low concentration SDS decellularised porcine dermis

The aim of this study was to adapt a proprietary decellularisation process for human dermis for use with porcine skin. Porcine skin was subject to: sodium chloride (I M) to detach the epidermis, trypsin paste to remove hair follicles, peracetic acid (0.1% v/v) disinfection, washed in hypotonic buffer and 0.1% (w/v) sodium dodecyl sulphate in the presence of proteinase inhibitors followed by nuclease treatment. Cellular porcine skin, decellularised porcine and human dermis were compared using histology, immunohistochemistry, GSL-1 lectin (alpha-gal epitope) staining, biochemical assays, uniaxial tensile and in vitro cytotoxicity tests. There was no microscopic evidence of cells in decellularised porcine dermis. DNA content was reduced by 98.2 % compared to cellular porcine skin. There were no significant differences in the biomechanical parameters studied or evidence of cytotoxicity. The decellularised porcine dermis retained residual alpha-gal epitope. Basement membrane collagen IV immunostaining was lost following decellularisation however laminin staining was retained

Fluidic haptic interface for mechano-tactile feedback

Notable advancements have been achieved in providing amputees with sensation through invasive and non-invasive haptic feedback systems such as mechano-, vibro-, electrotactile and hybrid systems. Purely mechanical-driven feedback approaches, however, have been little explored. In this paper, we now created a haptic feedback system that does not require any external power source (such as batteries) or other electronic components. The system is low-cost, lightweight, adaptable and robust against external impact (such as water). Hence, it will be sustainable in many aspects. We have made use of latest multimaterial 3D printing technology (Stratasys Objet500 Connex3) being able to fabricate a soft sensor and a mechano-tactile feedback actuator made of a rubber (TangoBlack Plus) and plastic (VeroClear) material. When forces are applied to the fingertip sensor, fluidic pressure inside the system acts on the membrane of the feedback actuator resulting in mechano-tactile sensation. We present the design, fabrication and validation of the proposed haptic feedback system. Our ∅7 mm feedback actuator is able to transmit a force range between 0.2 N (the median touch threshold) and 2.1 N (the maximum force transmitted by the feedback actuator at a 3 mm indentation) corresponding to force range exerted to the fingertip sensor of 1.2 − 18.49 N

“Affimer” synthetic protein scaffolds block oxidized LDL binding to the LOX-1 scavenger receptor and inhibit ERK1/2 activation

In multicellular organisms, a variety of lipid-protein particles control the systemic flow of triacylglycerides, cholesterol and fatty acids between cells in different tissues. The chemical modification by oxidation of these particles can trigger pathological responses, mediated by a group of membrane proteins termed scavenger receptors. The lectin-like oxidised low-density lipoprotein (LOX-1) scavenger receptor binds to oxidized low density lipoprotein (oxLDL) and mediates both signaling and trafficking outcomes. Here, we identified 5 synthetic proteins termed Affimers from a phage display library, each capable of binding recombinant LOX-1 extracellular (oxLDL-binding) domain with high specificity. These Affimers, based on a phytocystatin scaffold with loop regions of variable sequence, were able to bind to the plasma membrane of HEK293T cells exclusively when human LOX-1 was expressed. Binding and uptake of fluorescently-labelled oxLDL by the LOX-1-expressing cell model was inhibited with sub-nanomolar potency by all 5 Affimers. ERK1/2 activation, stimulated by oxLDL binding to LOX-1, was also significantly inhibited (p<0.01) by pre-incubation with LOX-1-specific Affimers, but these Affimers had no direct agonistic effect. Molecular modeling indicated that the LOX-1-specific Affimers bound predominantly via their variable loop regions to the surface of the LOX-1 lectin-like domain that contains a distinctive arrangement of arginine residues previously implicated in oxLDL binding, involving interactions with both subunits of the native, stable scavenger receptor homodimer. These data provide a new class of synthetic tools to probe and potentially modulate the oxLDL/LOX-1 interaction that plays an important role in vascular disease

Real-Time In Vivo Imaging of Early Mucosal Changes during Ischemia-Reperfusion in Human Jejunum

BACKGROUND AND STUDY AIMS: Small intestinal ischemia-reperfusion (IR) is a frequent, potentially life threatening phenomenon. There is a lack of non-invasive diagnostic modalities. For many intestinal diseases, visualizing the intestinal mucosa using endoscopy is gold standard. However, limited knowledge exists on small intestinal IR-induced, early mucosal changes. The aims of this study were to investigate endoscopic changes in human jejunum exposed to IR, and to study concordance between endoscopic appearance and histology. PATIENTS AND METHODS: In 23 patients a part of jejunum, to be removed for surgical reasons, was isolated and selectively exposed to ischemia with 0, 30 or 120 minutes of reperfusion. In 3 patients, a videocapsule was inserted in the isolated segment before exposure to IR, to visualize the mucosa. Endoscopic view at several time points was related to histology (Heamatoxylin & Eosin) obtained from 20 patients. RESULTS: Ischemia was characterized by loss of villous structure, mucosal whitening and appearance of punctate lesions. This was related to appearance of subepithelial spaces and breaches in the epithelial lining in the histological view. Early during reperfusion, the lumen filled with IR-damaged, shed cells and VCE showed mucosal erosions, hemorrhage and intraluminal debris. At 60 minutes of reperfusion, the only remaining signs of IR were loss of villous structure and small erosions, indicating rapid mucosal healing. CONCLUSIONS: This study shows a unique, real-time in vivo endoscopic view of early mucosal changes during IR of the human small intestine. Future studies should evaluate its usefulness in diagnosis of patients suspected of IR

In silico design and biological evaluation of a dual specificity kinase inhibitor targeting cell cycle progression and angiogenesis

Methodology: We have utilized a rational in silico-based approach to demonstrate the design and study of a novel compound that acts as a dual inhibitor of vascular endothelial growth factor receptor 2 (VEGFR2) and cyclin-dependent kinase 1 (CDK1). This compound acts by simultaneously inhibiting pro-Angiogenic signal transduction and cell cycle progression in primary endothelial cells. JK-31 displays potent in vitro activity against recombinant VEGFR2 and CDK1/cyclin B proteins comparable to previously characterized inhibitors. Dual inhibition of the vascular endothelial growth factor A (VEGF-A)-mediated signaling response and CDK1-mediated mitotic entry elicits anti-Angiogenic activity both in an endothelial-fibroblast co-culture model and a murine ex vivo model of angiogenesis