Robust envelope exchange platform for oncolytic measles virus.

The use of oncolytic viruses (OV) to precisely target and eliminate tumors ('virotherapy') is a rapidly evolving therapeutic approach to treating cancer. A major obstacle in virotherapy, especially for systemic administration, is the host's immune response towards the OV. In the case of measles virus (MeV), most individuals have been immunized against this agent leading to pre-existing neutralizing antibodies that can impair OV delivery to the tumor. These antibodies predominantly target the hemagglutinin (H) and fusion (F) envelope glycoproteins displayed at the particle's surface. Here, we introduce a novel and versatile pseudotyping platform for rapid envelope exchange of oncolytic MeV that allows for engineering of chimeric viruses invulnerable to pre-existing anti-MeV antibodies. Using this system, we have successfully exchanged the MeV F and H proteins with the glycoprotein G of vesicular stomatitis virus (VSV) and the surface proteins of Newcastle disease virus (NDV) or canine distemper virus (CDV), all of which are not endemic in the general human population. While the MeV-VSV and MeV-NDV pseudotypes were non-functional, the MeV-CDV pseudotype was successfully propagated to high-titer virus stocks. This study describes the successful generation of a robust envelope exchange platform for oncolytic MeV while also highlighting its intricate pseudotyping tolerance.

The association between genome-wide polymorphisms and chronic postoperative pain: a prospective observational study.

Chronic postoperative pain is common and can have a negative impact on quality of life. Recent studies show that genetic risk factors are likely to play a role, although only gene-targeted analysis has been used to date. This is the first genome-wide association study to identify single-nucleotide polymorphisms associated with the development of chronic postoperative pain based on two independent cohorts. In a discovery cohort, 330 women scheduled for hysterectomy were genotyped. A case-control association analysis compared patients without chronic postoperative pain and the 34 who had severe chronic postoperative pain 3 months after surgery. No single-nucleotide polymorphisms reached genome-wide significance, but several showed suggestive associations with chronic postoperative pain (p < 1 × 10-5 ). Single-nucleotide polymorphisms with significance p < 1 × 10-5 were followed up in a replication cohort consisting of 203 men and women scheduled for orthopaedic or abdominal surgery. Ten of these patients developed severe chronic postoperative pain. A single-nucleotide polymorphism in NAV3 was significantly replicated with chronic postoperative pain in the replication cohort (p = 0.009). Meta-analysis revealed that two loci (IQGAP1 and CRTC3) were significantly associated with chronic postoperative pain at 3 months (IQGAP1 p = 3.93 × 10-6 ß = 2.3863, CRTC3 p = 2.26 × 10-6 , ß = 2.4209). The present genome-wide association study provides initial evidence for genetic risk factors of chronic postoperative pain and supports follow-up studies.

Reusable High Aspect Ratio 3-D Nickel Shadow Mask.

Shadow Mask technology has been used over the years for resistless patterning and to pattern on unconventional surfaces, fragile substrate and biomaterial. In this work, we are presenting a novel method to fabricate high aspect ratio (15:1) three-dimensional (3D) Nickel (Ni) shadow mask with vertical pattern length and width of 1.2 mm and 40 µm respectively. The Ni shadow mask is 1.5 mm tall and 100 µm wide at the base. The aspect ratio of the shadow mask is 15. Ni shadow mask is mechanically robust and hence easy to handle. It is also reusable and used to pattern the sidewalls of unconventional and complex 3D geometries such as microneedles or neural electrodes (such as the Utah array). The standard Utah array has 100 active sites at the tip of the shaft. Using the proposed high aspect ratio Ni shadow mask, the Utah array can accommodate 300 active sites, 200 of which will be along and around the shaft. The robust Ni shadow mask is fabricated using laser patterning and electroplating techniques. The use of Ni 3D shadow mask will lower the fabrication cost, complexity and time for patterning out-of-plane structures.

Long term performance of porous platinum coated neural electrodes.

Over the last several years, there has been a growing interest in neural implants for the study and diagnostics of neurological disorders as well as for the symptomatic treatment of central nervous system related diseases. One of the major challenges is the trade-off between small electrode sizes for high selectivity between single neurons and large electrode-tissue interface areas for excellent stimulation and recording properties. This paper presents an approach of increasing the real surface area of the electrodes by creating a surface microstructure. Two major novelties let this work stand out from existing approaches which mainly make use of porous coatings such as platinum black or iridium oxide, or Poly(3,4-ethylenedioxythiophene) (PEDOT). Roughening is carried out by a dry etching process on the silicon electrode core before being coated by a sputtered platinum layer, eliminating complicated deposition processes as for the materials described above. The technology is compatible with any commonly used coating material. In addition, the surface roughening is compatible with high aspect ratio penetrating electrode arrays such as the well-established Utah electrode array, whose unique geometry presents a challenge in the surface modification of active electrode sites. The dry etching process is well characterized and yields a high controllability of pore size and depth. This paper confirms the superior electrochemical properties including impedance, charge injection capacity, and charge storage capacity of surface engineered electrode arrays compared to conventional arrays over a period of 12 weeks. Furthermore, mechanical stability of the modified electrodes was tested by implantation in the brain of a recently deceased rat. In conclusion, the larger interface surface of the electrodes does not only decrease the impedance which should lead to enhanced Signal to noise ratio (SNR) for recording purposes, but also yields higher charge injection capacities, which improve the stimulation characteristics of the implants.

Different methods to alter surface morphology of high aspect ratio structures.

In various applications such as neural prostheses or solar cells, there is a need to alter the surface morphology of high aspect ratio structures so that the real surface area is greater than geometrical area. The change in surface morphology enhances the devices functionality. One of the applications of altering the surface morphology is of neural implants such as the Utah electrode array (UEA) that communicate with single neurons by charge injection induced stimulation or by recording electrical neural signals. For high selectivity between single cells of the nervous system, the electrode surface area is required to be as small as possible, while the impedance is required to be as low as possible for good signal to noise ratios (SNR) during neural recording. For stimulation, high charge injection and charge transfer capacities of the electrodes are required, which increase with the electrode surface. Traditionally, researchers have worked with either increasing the roughness of the existing metallization (Platinum grey, black) or other materials such as Iridium Oxide and PEDOT. All of these previously investigated methods lead to more complicated metal deposition processes that are difficult to control and often have a critical impact on the mechanical properties of the metal films. Therefore, a modification of the surface underneath the electrode's coating will increase its surface area while maintaining the standard and well controlled metal deposition process. In this work, the surfaces of the Silicon micro-needles were engineered by creating a defined microstructure on the electrodes surface using several methods such as Laser ablation, focused ion beam, sputter etching, reactive ion etching (RIE) and deep reactive ion etching (DRIE). The surface modification processes were optimized for the high aspect ratio Silicon structures of the UEA. The increase in real surface area while maintaining the geometrical surface area was verified using scanning electron microscopy (SEM) and electrochemical impedance spectroscopy (EIS). The best results were obtained by DRIE induced surface morphology. Decreases in impedance values of electrodes up to 76 % indicate the successful surface engineering of the high aspect ratio Silicon structures.

Genome-wide analysis implicates microRNAs and their target genes in the development of bipolar disorder.

Bipolar disorder (BD) is a severe and highly heritable neuropsychiatric disorder with a lifetime prevalence of 1%. Molecular genetic studies have identified the first BD susceptibility genes. However, the disease pathways remain largely unknown. Accumulating evidence suggests that microRNAs, a class of small noncoding RNAs, contribute to basic mechanisms underlying brain development and plasticity, suggesting their possible involvement in the pathogenesis of several psychiatric disorders, including BD. In the present study, gene-based analyses were performed for all known autosomal microRNAs using the largest genome-wide association data set of BD to date (9747 patients and 14 278 controls). Associated and brain-expressed microRNAs were then investigated in target gene and pathway analyses. Functional analyses of miR-499 and miR-708 were performed in rat hippocampal neurons. Ninety-eight of the six hundred nine investigated microRNAs showed nominally significant P-values, suggesting that BD-associated microRNAs might be enriched within known microRNA loci. After correction for multiple testing, nine microRNAs showed a significant association with BD. The most promising were miR-499, miR-708 and miR-1908. Target gene and pathway analyses revealed 18 significant canonical pathways, including brain development and neuron projection. For miR-499, four Bonferroni-corrected significant target genes were identified, including the genome-wide risk gene for psychiatric disorder CACNB2. First results of functional analyses in rat hippocampal neurons neither revealed nor excluded a major contribution of miR-499 or miR-708 to dendritic spine morphogenesis. The present results suggest that research is warranted to elucidate the precise involvement of microRNAs and their downstream pathways in BD.

Single-Electron Detection and Spectroscopy via Relativistic Cyclotron Radiation.

It has been understood since 1897 that accelerating charges must emit electromagnetic radiation. Although first derived in 1904, cyclotron radiation from a single electron orbiting in a magnetic field has never been observed directly. We demonstrate single-electron detection in a novel radio-frequency spectrometer. The relativistic shift in the cyclotron frequency permits a precise electron energy measurement. Precise beta electron spectroscopy from gaseous radiation sources is a key technique in modern efforts to measure the neutrino mass via the tritium decay end point, and this work demonstrates a fundamentally new approach to precision beta spectroscopy for future neutrino mass experiments.

MicroRNA-mediated multi-tissue detargeting of oncolytic measles virus.

Precise oncotropism is required for successful systemic administration of next-generation oncolytic measles viruses (MVs). We have previously established a system for efficient post-entry targeting by insertion of synthetic microRNA target sites (miRTS) into the MV genome, thereby repressing replication in the presence of cognate microRNAs. Thus, differential expression of microRNAs, as frequently observed in normal compared with malignant tissues, can be exploited to increase vector specificity and safety. Here we report the combination of miRTS for different microRNAs in a single vector to detarget pivotal organs at risk during systemic administration (liver, brain, gastrointestinal tract). Accordingly, miRTS for miR-122, miR-7 and miR-148a that are enriched in these tissues were inserted to create multi-tissue-detargeted MV (MV-EGFP(mtd)). Replication of MV-EGFP(mtd) is repressed in cell lines as well as in non-transformed primary human hepatocytes and liver slices expressing cognate microRNAs. Oncolytic potency of MV-EGFP(mtd) is retained in a model of pancreatic cancer in vitro and in vivo. This work is a proof-of-concept that favorable expression profiles of multiple microRNAs can be exploited concomitantly to reshape the tropism of MV without compromising oncolytic efficacy. This strategy can be adapted to different vectors and cancer entities for safe and efficient high-dose systemic administration in clinical trials.

Follow-up of loci from the International Genomics of Alzheimer's Disease Project identifies TRIP4 as a novel susceptibility gene.

To follow-up loci discovered by the International Genomics of Alzheimer's Disease Project, we attempted independent replication of 19 single nucleotide polymorphisms (SNPs) in a large Spanish sample (Fundació ACE data set; 1808 patients and 2564 controls). Our results corroborate association with four SNPs located in the genes INPP5D, MEF2C, ZCWPW1 and FERMT2, respectively. Of these, ZCWPW1 was the only SNP to withstand correction for multiple testing (P=0.000655). Furthermore, we identify TRIP4 (rs74615166) as a novel genome-wide significant locus for Alzheimer's disease risk (odds ratio=1.31; confidence interval 95% (1.19-1.44); P=9.74 × 10(-)(9)).

Chemovirotherapy for head and neck squamous cell carcinoma with EGFR-targeted and CD/UPRT-armed oncolytic measles virus.

First-line treatment of recurrent and/or refractory head and neck squamous cell carcinoma (HNSCC) is based on platinum, 5-fluorouracil (5-FU) and the monoclonal antiEGFR antibody cetuximab. However, in most cases this chemoimmunotherapy does not cure the disease, and more than 50% of HNSCC patients are dying because of local recurrence of the tumors. In the majority of cases, HNSCC overexpress the epidermal growth factor receptor (EGFR), and its presence is associated with a poor outcome. In this study, we engineered an EGFR-targeted oncolytic measles virus (MV), armed with the bifunctional enzyme cytosine deaminase/uracil phosphoribosyltransferase (CD/UPRT). CD/UPRT converts 5-fluorocytosine (5-FC) into the chemotherapeutic 5-FU, a mainstay of HNSCC chemotherapy. This virus efficiently replicates in and lyses primary HNSCC cells in vitro. Arming with CD/UPRT mediates efficient prodrug activation with high bystander killing of non-infected tumor cells. In mice bearing primary HNSCC xenografts, intratumoral administration of MV-antiEGFR resulted in statistically significant tumor growth delay and prolongation of survival. Importantly, combination with 5-FC is superior to virus-only treatment leading to significant tumor growth inhibition. Thus, chemovirotherapy with EGFR-targeted and CD/UPRT-armed MV is highly efficacious in preclinical settings with direct translational implications for a planned Phase I clinical trial of MV for locoregional treatment of HNSCC.

Armed and targeted measles virus for chemovirotherapy of pancreatic cancer.

No curative therapy is currently available for locally advanced or metastatic pancreatic cancer. Therefore, new therapeutic approaches must be considered. Measles virus (MV) vaccine strains have shown promising oncolytic activity against a variety of tumor entities. For specific therapy of pancreatic cancer, we generated a fully retargeted MV that enters cells exclusively through the prostate stem cell antigen (PSCA). Besides a high-membrane frequency on prostate cancer cells, this antigen is expressed on pancreatic adenocarcinoma, but not on non-neoplastic tissue. PSCA expression levels differ within heterogeneous tumor bulks and between human pancreatic cell lines, and we could show specific infection of pancreatic adenocarcinoma cell lines with both high- and low-level PSCA expression. Furthermore, we generated a fully retargeted and armed MV-PNP-anti-PSCA to express the prodrug convertase purine nucleoside phosphorylase (PNP). PNP, which activates the prodrug fludarabine effectively, enhanced the oncolytic efficacy of the virus on infected and bystander cells. Beneficial therapeutic effects were shown in a pancreatic cancer xenograft model. Moreover, in the treatment of gemcitabine-resistant pancreatic adenocarcinoma cells, no cross-resistance to both MV oncolysis and activated prodrug was detected.

[Balloon dacryocystoplasty: interventional radiological therapy of congenital dacryostenosis]. / Ballondakryozystoplastie: Interventionell-radiologische Therapie konnataler Dakryostenosen.

PURPOSE: To evaluate the results of balloon dacryocystoplasty in the treatment of complicated development of connatal obstructed nasolacrimal duct system. MATERIALS AND METHODS: Dacryocystography under general anaesthesia was performed on 46 children with epiphora from birth and recurrent infection of the nasolacrimal duct system. 54 nasolacrimal ducts (8 children bilaterally) were treated with balloon catheter dilatation and antibiotic irrigation of the nasolacrimal sac. In all cases previous conservative treatment with eye drops and superficial massage of the lacrimal sac had failed. 11 children without clinical improvement were irrigated before catheter dilatation by an ophthalmologist. The ages ranged from 6 weeks to 7.5 years (mean 23.5 months). 39 dilatations were carried out as an out-patient procedure. The clinical results were confirmed by a questionnaire filled in by the parents. RESULTS: 15 incomplete obstructions and 39 occlusions of the Hasner valve (n = 45) or of the nasolacrimal duct system (n = 9) were demonstrated with dacryocystography. Dilatation with a 2.5 mm ballon was successfully performed in all cases and the mean radiation time was 2.1 minutes. No relevant complications occurred. The mean follow up time was 18.4 months (3 - 41 months). 39 of 46 children showed no symptoms, 6 children seldomly experienced onset of epiphora. The symptoms did not improve in only one child. The cumulative clinical success rate is 98 %. CONCLUSION: Following diagnostic dacryocystography, balloon catheter dilatation is a low risk and very successful treatment of complicated connatal obstructed nasolacrimal duct system.

A standardized assessment and intervention protocol for managing risk for falls on a geriatric rehabilitation unit.

This article describes the intervention protocols implemented at the Sisters of Charity Hospital of Ottawa, Canada on a 36-bed geriatric rehabilitation unit for managing the risk for falls in patients assessed to be at high, moderate, or low risk. The implementation of tailored protocols is of particular importance on rehabilitation units because staff must balance the treatment goal to increase patient mobility, activity level, and independence with the need to prevent falls. Thus, this article also describes the related standardized criteria and decisional guidelines used in determining patients' level of independence on the unit for each fall-risk category. This article concludes with an outline of practices designed to facilitate safe transfers and completion of activities of daily living among geriatric patients with functional impairments by teaching and promoting the use of behavioral compensatory strategies for disability-related deficits.

Stereotactic biopsy of intracranial brain lesions. High diagnostic yield without increased complications: 65 consecutive biopsies with early postoperative CT scans.

Diagnostic yield and complication rate were analyzed for a series of 65 consecutive stereotactic biopsies of intra-axial brain lesions. The diagnostic yield was 98.5 +/- 1.5% and the complication rate was 1.5%. A median number of 14 biopsies (range 1-48) were taken per lesion. The biopsy sites followed a clockwise pattern, going from the superficial margin to the center and the deep margin of the lesion with respect to the inner table of the skull. A side window cannula biopsy needle was used. All patients underwent immediate postoperative CT scans within 4 h of biopsy to rule out intracranial complications. All patients were discharged within 24 h after biopsy, unless medical reasons unrelated to the biopsy required further hospitalization. We attribute the high diagnostic yield in our series to the high number of systematically taken biopsies per lesion. The higher number of biopsies did not lead to an increase in complications. From our experience, it appears safe to discharge patients the same day or within 24 h after a stereotactic biopsy if the postoperative CT shows no complication. Stereotactic biopsy could often safely be performed on an outpatient basis.

[Catheterization of the nasolacrimal duct]. / Sondierung des Ductus nasolacrimalis.

A technique is described for passing a guide wire through the lacrimal duct into the nose following catheterisation of the duct. The wire is passed into the pharynx, a 7 F feeding tube is passed through the nose into the pharynx and these are then pulled out of the mouth with a Magill forceps. The tip of the feeding tube is then cut off and the guide wire introduced into a side hole of the catheter. By removing the catheter, the tip of the guide wire is pulled along and can be removed from the nose.

Aspects of cognitive status as predictors of mobility following geriatric rehabilitation.

This study aimed to determine the specific aspects of cognitive functioning which are related to geriatric rehabilitation treatment-outcome, and was based on a within-subject repeated measures design. In the setting of the inpatient Geriatric Rehabilitation Program (GRP) of the SCO Hospital in Ottawa, Canada 40 patients underwent six weeks of physiotherapy treatment for mobility training, on a twice daily schedule. Measurements were performed by the Clinical Outcome Variables Scale (COVS) and a mental status battery composed of five neuropsychological tests. Patients with a poorer mobility status at admission were significantly more depressed, more apraxic, less educated and had greater memory problems than their counterparts. The extent of treatment gains achieved by discharge was significantly related to a single aspect of cognition, namely procedural memory. Longer-term maintenance of treatment gains, however, was predicted by cognitive functioning more globally, including measures of praxis, declarative memory and reasoning. Together, the measures of cognition explained 52% of the variance in functional mobility outcome. In conclusion, patients with mild-to-moderate difficulties of cortically-based higher-order cognitive functions may still achieve significant gains in mobility function following geriatric rehabilitation. However, these patients may not be able to maintain their gains over time to the same extent as their cognitively healthier counterparts. Thus, from an empirical viewpoint, the decision to include or exclude cognitively impaired patients in GRPs varies with the definition of "treatment success" selected (i.e., short-term vs long-term gains). Alternatively, cognitively impaired patients may require more frequent follow-up rehabilitation services in order to maintain their gains.

N-acetylglucose aminidase activity in corneoscleral ulceration after severe eye burns.

In some patients with severe burns, major problems were delayed regeneration of the surface epithelium, extensive ulceration of the sclera near the limbus, subsequent corneal ulceration, and subconjunctival scarring. Human tissues surrounding ulcerations in severe eye burns were obtained from surgical interventions in 12 such patients. High activities of the lysosomal marker enzyme N-acetyl-glucose aminidase were found. As is known from histological examinations, these tissues do not represent regeneration of conjunctiva, but rather inflammatory proliferation. The tissues adjacent to corneoscleral ulceration release large amounts of lysosomal destructive enzymes. The rather simple assay of the activity of the N-acetylglucose aminidase proved useful in clinical cases to assess the activity of the inflammation of eye burns and to estimate the efficacy of the therapy applied.

Self-mutilation: three cases and a review of the literature.

The self-mutilating patient is an unusual psychiatric presentation in the emergency department (ED). Nonetheless, serious underlying psychiatric pathology and drug abuse are important background risk factors. A careful stepwise approach in the ED is essential, although the prognosis, follow-up, and eventual rehabilitation are poor.

Effect of epidermal growth factor in severe experimental alkali burns.

Ten rabbits with the right eye burnt by 1 N NaOH were treated 5 times daily with epidermal growth factor (EGF) eyedrops or placebo. The epithelium seemed to heal better under EGF treatment than with placebo. Also, ulceration and secondary calcification of the cornea were reduced in the EGF-treated group. The vascularisation was slightly diminished by EGF. The lactate dehydrogenase and N-acetylglucose aminidase activities and the lactate and glucose levels in the corneae were not different between the EGF and the placebo groups. The results showed that there was a beneficial effect of EGF in these experiments with most severe eye burn disease. But the improved regeneration of the epithelium seemed to be counteracted and partly abolished by the severe inflammatory reaction in these eyes.