Synthesis of Novel Antimicrobial CHX-CaCl2 Coatings on Maxillofacial Fixatures for Infection Prevention.

Maxillofacial surgery placement of fixatures (Leonard Buttons, LB) at close proximity to surgical incisions provides a potential reservoir as a secondary local factor to advanced periodontal disease, with bacterial formation around failed fixatures implicating plaque. To address infection rates, we aimed to surface coat LB and Titanium (Ti) discs using a novel form of chlorhexidine (CHX), CHX-CaCl2 and 0.2% CHX digluconate mouthwash as a comparison. CHX-CaCl2 coated, double-coated and mouthwash coated LB and Ti discs were transferred to 1 mL artificial saliva (AS) at specified time points, and UV-Visible spectroscopy (254 nm) was used to measure CHX release. The zone of inhibition (ZOI) was measured using collected aliquots against bacterial strains. Specimens were characterized using Energy Dispersive X-ray Spectroscopy (EDS), X-ray Diffraction (XRD) and Scanning Electron Microscopy (SEM). SEM displayed copious dendritic crystals on LB/ Ti disc surfaces. Drug release from double-coated CHX-CaCl2 was 14 days (Ti discs) and 6 days (LB) above MIC, compared to the comparison group (20 min). The ZOI for the CHX-CaCl2 coated groups was significantly different within groups (p < 0.05). CHX-CaCl2 surface crystallization is a new drug technology for controlled and sustained CHX release; its antibacterial effectiveness makes this drug an ideal adjunct following clinical and surgical procedures to maintain oral hygiene and prevent surgical site infections.

Investigation of the allele frequency of the G>A intron 10 ADAMTS17 mutation causing primary lens luxation in the Portuguese Podengo breed.

OBJECTIVE: To establish the allele frequency of the PLL-causing G>A intron 10 ADAMTS17 mutation in the Portuguese Podengo population in the UK and investigate a possible correlation between the mutation and short stature. METHODS: Two groups of dogs (Group 1 and Group 2) were recruited for the purpose of the study. Group 1 (n = 40) consisted of dogs which were genotyped only and Group 2 (n = 42) consisted of dogs which were genotyped, underwent a full ophthalmological examination and also had their height measured at the withers. RESULTS: In Group 1, genotyping for the ADAMTS17:c.1473+1G>A mutation confirmed 1/40 homozygous for the mutated allele (-/-), 7/40 heterozygous for the mutated allele (+/-), and 32/40 homozygous for the wild-type allele (+/+) dogs. In Group 2, genotyping of the dogs confirmed 6/42 heterozygous for the mutated allele (+/-) and homozygous for the wild-type allele (+/+) dogs. In total, 1/82 (1.2%) dogs were confirmed to be homozygous for the mutated allele, 13/82 (15.8%) heterozygous for the mutated allele and 68/82 (83%) homozygous for the wild-type allele. The frequency of the mutated allele across both groups was calculated as 0.09. A statistically significant correlation between the mutation and short stature could not be established (p = .590). CONCLUSIONS: The frequency of the mutation calculated in this study (0.09) is high. Genetic testing should be considered for each dog prior to breeding with a view of selective breeding.

Melting corneal ulcers (keratomalacia) in dogs: A 5-year clinical and microbiological study (2014-2018).

OBJECTIVES: To identify bacterial microorganisms associated with canine keratomalacia, review their antimicrobial sensitivity, and evaluate clinical outcomes compared to results of microbial culture. METHODS: Retrospective analysis of clinical records of dogs diagnosed with a melting corneal ulcer presented to a referral hospital in Hertfordshire, UK between 2014 and 2018. RESULTS: One hundred and ten melting corneal ulcers were sampled in 106 dogs. The most common pure bacterial isolate was Pseudomonas aeruginosa (n = 26) followed by ß-hemolytic Streptococcus (n = 12). Melting corneal ulcers that cultured coagulase-positive Staphylococcus, coliform bacteria, Pasteurella multocida, Enterococcus, and Streptococcus viridans presented in smaller numbers and were analyzed together (n = 16). Multiple cultures were identified in nine cases (n = 9). Forty-seven cultures yielded no bacterial growth (n = 47). The susceptibility to fluoroquinolones remained high with the exception of ß-hemolytic Streptococci. There was no significant difference in the ulcer severity at presentation in regard to the cultured bacteria. Overall, 63 eyes (57%) received surgical grafting in addition to medical treatment. In 14 cases (13%), the progression of corneal melting despite medical ± surgical treatment resulted in enucleation. Fifty-seven percent (8/14) of the enucleated eyes cultured pure Pseudomonas aeruginosa isolates. In contrast, all ß-hemolytic Streptococcus-associated ulcers healed. CONCLUSIONS: The most common bacterial species associated with canine keratomalacia were Pseudomonas aeruginosa and ß-hemolytic Streptococcus. Because of the variation in antibacterial sensitivity between these two species, bacterial culture and sensitivity testing should be performed in all dogs presenting with keratomalacia. Melting corneal ulcers associated with pure Pseudomonas infection were significantly more likely to result in globe loss than melting corneal ulcers associated with other cultures.

Immediate effects of diamond burr debridement in patients with spontaneous chronic corneal epithelial defects, light and electron microscopic evaluation.

PURPOSE: To evaluate immediate effects of diamond burr debridement (DBD) on the cornea of canine patients diagnosed with spontaneous chronic corneal epithelial defects (SCCEDs). ANIMALS STUDIED: Eight client owned dogs with SCCEDs. METHODS: Nine eyes from eight dogs with SCCEDs underwent superficial keratectomy (SK). The ulcerated area was divided into quadrants with a 300-micron restricted depth knife. Two of four quadrants underwent DBD for 40-60 s. A SK followed immediately. One burred section and one nonburred section were fixed with formaldehyde 10% and underwent light microscopy (LM). The remaining quadrants from five eyes were fixed with glutaraldehyde 2.5% and underwent transmission electron microscopy (TEM). Masked pathologists evaluated the samples. A student's paired t-test was used to analyze the data. RESULTS: With LM all nonburred samples had a superficial stromal hyaline acellular zone (HAZ), seven of the burred samples had an intermittent HAZ and in two burred samples this zone was absent. The HAZ thickness of burred samples (1.062 ± 0.664 µm) was significantly thinner than that of the nonburred samples (4.309 ± 1.348 µm) (P < 0.0001). Transmission electron microscopy showed an absence of basement membrane and the presence of an amorphous, fine fibrillar material in the superficial stroma in nonburred samples. This material was intermittent or absent in burred samples. CONCLUSION: DBD significantly reduces the superficial stromal HAZ in SCCEDs. A reduction of its thickness may be responsible for the healing rates reported with DBD.

Cadaveric experiments to evaluate pressure wave generated by radial shockwave treatment of plantar fasciitis.

BACKGROUND: Shockwave treatment is increasingly used for plantar fasciitis and Achilles tendinopathy. To be effective it is believed that high pressure must be achieved in the tissues. We report on the first human cadaveric experiments to characterize pressure from radial shockwave therapy (rSWT) for plantar fasciitis. METHODS: The pressure from rSWT was measured in two cadaveric feet using a needle hydrophone. Maximal pressure and energy flux were calculated from the measurements. RESULTS: The pressure persisted longer than supposed, for up to 400µs. The peak negative pressure was up to two Mega Pascal. The predicted energy in the tissue strongly depended on the time interval used in calculations. CONCLUSIONS: The measured pressure may be sufficiently high to cause cavitation in the tissue, which is one of the proposed healing mechanisms associated with rSWT. The results suggest that the energy is imparted to the tissues for much longer than previously thought.

Local and Sustained Activity of Doxycycline Delivered with Layer-by-Layer Microcapsules.

Achieving localized delivery of small molecule drugs has the potential to increase efficacy and reduce off target and side effects associated with systemic distribution. Herein, we explore the potential use of layer-by-layer (LbL) assembled microcapsules for the delivery of doxycycline. Absorbance of doxycycline onto core dextran sulfate of preassembled microcapsules provides an efficient method to load both synthetic and biodegradable microcapsules with the drug. Application of an outer layer lipid coat enhances the sustained in vitro release of doxycycline from both microcapsule types. To monitor doxycycline delivery in a biological system, C2C12 mouse myoblasts are engineered to express EGFP under the control of the optimized components of the tetracycline regulated gene expression system. Microcapsules are not toxic to these cells, and upon delivery to the cells, EGFP is more efficiently induced in those cells that contain engulfed microcapsules and monitored EGFP expression clearly demonstrates that synthetic microcapsules with a DPPC coat are the most efficient for sustain intracellular delivery. Doxycycline released from microcapsules also displayed sustained activity in an antimicrobial growth inhibition assay compared with doxycycline solution. This study reveals the potential for LbL microcapsules in small molecule drug delivery and their feasible use for achieving prolonged doxycycline activity.

Survey of the incidence of pectinate ligament dysplasia and glaucoma in the UK Leonberger population.

OBJECTIVE: To determine the prevalence of pectinate ligament dysplasia (PLD) in UK Leonbergers and identify cases affected by glaucoma. Also, to define the spectrum of pectinate ligament (PL) appearance in this breed and determine whether gonioscopic monitoring should be recommended. ANIMALS STUDIED: Data were compiled from 78 prospective gonioscopy examinations performed by one author (GF) and retrospective analysis of 233 UK eye scheme certificates (2009-2014). Clinical cases of glaucoma in Leonbergers diagnosed by UK veterinary ophthalmologists, where gonioscopy of the fellow eyes or histology of affected eyes had been performed, were also reviewed. PROCEDURE: In the prospective study, intraocular pressure was recorded prior to gonioscopy using a rebound tonometer. Gonioscopy was performed using a slit-lamp biomicroscope with a Koeppe goniolens. PLD was categorized according to the percentage of the iridocorneal drainage angle affected (grade 0 = <25% affected; grade 1 = 25-50% affected; grade 2 = 51-75% affected; and grade 3 = >75% affected), and the degree of narrowing of the angle was noted. RESULTS: Of 78 dogs examined prospectively, 64/78 (82%) were grade 0, 7/78 (9%) were grade 1, 3/78 (4%) were grade 2, and 4/78 (5%) were grade 3. A large phenotypic variation was observed. Spearman's rank correlation showed a positive correlation between age and severity of PLD (P < 0.0055). 52 (22%) of Leonbergers examined under the UK eye scheme 2009-2014 were affected by PLD. Five clinical cases of glaucoma were reviewed where gonioscopy had been performed and one where histology was performed. All individuals had grade 3 PLD with gonioscopy of the contralateral eye or severe goniodysgenesis with histological sections of the affected eye. CONCLUSION: This survey suggests the prevalence of PLD is sufficient to justify ongoing screening of Leonbergers.

Utilising psychophysical techniques to investigate the effects of age, typeface design, size and display polarity on glance legibility.

Psychophysical research on text legibility has historically investigated factors such as size, colour and contrast, but there has been relatively little direct empirical evaluation of typographic design itself, particularly in the emerging context of glance reading. In the present study, participants performed a lexical decision task controlled by an adaptive staircase method. Two typefaces, a 'humanist' and 'square grotesque' style, were tested. Study I examined positive and negative polarities, while Study II examined two text sizes. Stimulus duration thresholds were sensitive to differences between typefaces, polarities and sizes. Typeface also interacted significantly with age, particularly for conditions with higher legibility thresholds. These results are consistent with previous research assessing the impact of the same typefaces on interface demand in a simulated driving environment. This simplified methodology of assessing legibility differences can be adapted to investigate a wide array of questions relevant to typographic and interface designs. Practitioner Summary: A method is described for rapidly investigating relative legibility of different typographical features. Results indicate that during glance-like reading induced by the psychophysical technique and under the lighting conditions considered, humanist-style type is significantly more legible than a square grotesque style, and that black-on-white text is significantly more legible than white-on-black.

Generation of an efficiently secreted, cell penetrating NF-κB inhibitor.

Gene therapy is a powerful approach to treat disease locally. However, if the therapeutic target is intracellular, the therapeutic will be effective only in the cells where the therapeutic gene is delivered. We have engineered a fusion protein containing an intracellular inhibitor of the transcription factor NF-κB pathway that can be effectively secreted from producing cells. This fusion protein is cleaved extracellularly by metalloproteinases allowing release of a protein transduction domain (PTD) linked to the NF-κB inhibitor for translocation into neighboring cells. We show that engineered molecules can be efficiently secreted (>80%); are cleaved with matrix metalloprotease-1; inhibit NF-κB driven transcription in a biological assay with a human reporter cell line; and display significant inhibition in mouse paw inflammation models when delivered by lentivirus or secreting cells. No inhibition of NF-κB transcription or therapeutic effect was seen using molecules devoid of the PTD and NF-κB inhibitory domains. By creating a fusion protein with an endogenous secretion partner, we demonstrate a novel approach to efficiently secrete PTD-containing protein domains, overcoming previous limitations, and allowing for potent paracrine effects.

Identification of ZDHHC14 as a novel human tumour suppressor gene.

Genomic changes affecting tumour suppressor genes are fundamental to cancer. We applied SNP array analysis to a panel of testicular germ cell tumours to search for novel tumour suppressor genes and identified a frequent small deletion on 6q25.3 affecting just one gene, ZDHHC14. The expression of ZDHHC14, a putative protein palmitoyltransferase with unknown cellular function, was decreased at both RNA and protein levels in testicular germ cell tumours. ZDHHC14 expression was also significantly decreased in a panel of prostate cancer samples and cell lines. In addition to our findings of genetic and protein expression changes in clinical samples, inducible overexpression of ZDHHC14 led to reduced cell viability and increased apoptosis through the classic caspase-dependent apoptotic pathway and heterozygous knockout of ZDHHC14 increased [CORRECTED] cell colony formation ability. Finally, we confirmed our in vitro findings of the tumour suppressor role of ZDHHC14 in a mouse xenograft model, showing that overexpression of ZDHHC14 inhibits tumourigenesis. Thus, we have identified a novel tumour suppressor gene that is commonly down-regulated in testicular germ cell tumours and prostate cancer, as well as given insight into the cellular functional role of ZDHHC14, a potential protein palmitoyltransferase that may play a key protective role in cancer.

Gene therapy for autoimmune diseases: quo vadis?

Biological therapies using antibodies and cytokines are becoming widespread for the treatment of chronic inflammatory autoimmune diseases. However, these treatments have several limitations - such as expense, the need for repeated injections and unwanted side-effects - that can be overcome by genetic delivery. This review summarizes the ingenuity, sophistication and variety of gene-therapy approaches that have been taken in the design of therapeutic molecules and vectors, the engineering of cells and the regulation of gene expression for the targeting of disease outcome. We focus our attention on multiple sclerosis, type 1 diabetes and rheumatoid arthritis.

Progression of pectinate ligament dysplasia over time in two populations of Flat-Coated Retrievers.

OBJECTIVE: Two of the authors (DG, BS) independently observed that a number of Flat-Coated Retrievers (FCRs) previously unaffected by pectinate ligament dysplasia (PLD) appeared to develop the condition later in life. This study was instigated to investigate progression of PLD within individual dogs over time. ANIMALS STUDIED: Flat-Coated Retrievers that had previously undergone gonioscopy under the UK/ECVO hereditary eye schemes were included in the study. PROCEDURE: A second gonioscopic examination was performed 1.92-12.58 years later (mean 6, median 5.75 years) and the results compared. 39 FCR (17 males, 22 females) in the UK and 57 FCR (27 males, 30 females) in Switzerland were included. Slit-lamp biomicroscopy, indirect ophthalmoscopy, and gonioscopy were performed in all dogs. Gonioscopy allowed classification as either unaffected or affected; percentage of the iridocorneal drainage angle (ICA) affected by PLD was determined, before calculating progression observed as mild, moderate, or severe. RESULTS: 39 of 96 (40.6%) dogs demonstrated progression of PLD (P < 0.0001). Of these, 13 of 96 (13.5%) were classified as mild progression (from either unaffected to 10-20% or 10-20% to 20-90% ICA affected). Progression was more extensive in 26 of 96 (27.1%) dogs (P < 0.0001), of which 12 of 96 (12.5%) went from unaffected to severe PLD of >90% ICA affected, consistent with a high risk of glaucoma. CONCLUSIONS: To the authors' knowledge, this is the first report describing progression of PLD in individual dogs over time, in a breed affected by primary, angle closure glaucoma.

Validity of aqueocentesis as a component of anterior uveitis investigation in dogs and cats.

OBJECTIVE: To describe aqueocentesis cytopathology results from dogs and cats presenting for uveitis investigation and to determine whether this is a useful and safe procedure. ANIMAL STUDIED: Dogs and cats presenting for investigation of anterior uveitis (April 2008-December 2013). PROCEDURES: Aqueous was collected via limbal entry under sedation/general anesthesia, for cytopathology and occasionally bacterial culture or polymerase chain reaction (PCR) testing. Further workup included blood testing (hematology, biochemistry, and serology), diagnostic imaging, nonocular cytopathology, and available histopathology. RESULTS: Fifty-six dogs and 39 cats were included in the study. An aqueous cytopathologic diagnosis of lymphoma (or discrete cell neoplasia) was made in six dogs and seven cats, and a diagnosis of large cell carcinoma made in one dog. This diagnosis of lymphoma was confirmed by ocular histopathology in two dogs and one cat; nonocular cytopathology corroborated lymphoma in another three dogs and five cats. Lymphoma was not evident on aqueous cytopathology but confirmed on nonocular histopathology in two dogs and by cytopathology in one cat. Additionally, aqueous cytopathology in three cats suggested, but was not considered diagnostic of, lymphoma; one of these cats had a confirmatory diagnosis of lymphoma on subsequent clinical investigation. Aqueous humor cytopathology alone was not diagnostic in non-neoplastic anterior uveitis cases, but supplemented the clinical picture with other systemic diagnostic tests. No clinically important complications were reported in association with aqueocentesis. CONCLUSIONS: Aqueocentesis is performed readily with minimal risk. The results were primarily useful in aiding a diagnosis of lymphoma in both dogs and cats.

A comparative study of matrix metalloproteinase and aggrecanase mediated release of latent cytokines at arthritic joints.

BACKGROUND: Latent cytokines are engineered by fusing the latency associated peptide (LAP) derived from transforming growth factor-ß (TGF-ß) with the therapeutic cytokine, in this case interferon-ß (IFN-ß), via an inflammation-specific matrix metalloproteinase (MMP) cleavage site. OBJECTIVES: To demonstrate latency and specific delivery in vivo and to compare therapeutic efficacy of aggrecanase-mediated release of latent IFN-ß in arthritic joints to the original MMP-specific release. METHODS: Recombinant fusion proteins with MMP, aggrecanase or devoid of cleavage site were expressed in CHO cells, purified and characterised in vitro by Western blotting and anti-viral protection assays. Therapeutic efficacy and half-life were assessed in vivo using the mouse collagen-induced arthritis model (CIA) of rheumatoid arthritis and a model of acute paw inflammation, respectively. Transgenic mice with an IFN-regulated luciferase gene were used to assess latency in vivo and targeted delivery to sites of disease. RESULTS: Efficient localised delivery of IFN-ß to inflamed paws, with low levels of systemic delivery, was demonstrated in transgenic mice using latent IFN-ß. Engineering of latent IFN-ß with an aggrecanase-sensitive cleavage site resulted in efficient cleavage by ADAMTS-4, ADAMTS-5 and synovial fluid from arthritic patients, with an extended half-life similar to the MMP-specific molecule and greater therapeutic efficacy in the CIA model. CONCLUSIONS: Latent cytokines require cleavage in vivo for therapeutic efficacy, and they are delivered in a dose dependent fashion only to arthritic joints. The aggrecanase-specific cleavage site is a viable alternative to the MMP cleavage site for the targeting of latent cytokines to arthritic joints.

Assessing the impact of typeface design in a text-rich automotive user interface.

Text-rich driver-vehicle interfaces are increasingly common in new vehicles, yet the effects of different typeface characteristics on task performance in this brief off-road based glance context remains sparsely examined. Subjects completed menu selection tasks while in a driving simulator. Menu text was set either in a 'humanist' or 'square grotesque' typeface. Among men, use of the humanist typeface resulted in a 10.6% reduction in total glance time as compared to the square grotesque typeface. Total response time and number of glances showed similar reductions. The impact of typeface was either more modest or not apparent for women. Error rates for both males and females were 3.1% lower for the humanist typeface. This research suggests that optimised typefaces may mitigate some interface demands. Future work will need to assess whether other typeface characteristics can be optimised to further reduce demand, improve legibility, increase usability and help meet new governmental distraction guidelines. Practitioner Summary: Text-rich in-vehicle interfaces are increasingly common, but the effects of typeface on task performance remain sparsely studied. We show that among male drivers, menu selection tasks are completed with 10.6% less visual glance time when text is displayed in a 'humanist' typeface, as compared to a 'square grotesque'.

Process development for cell aggregate arrays encapsulated in a synthetic hydrogel using negative dielectrophoresis.

Spatial patterning of cells is of great importance in tissue engineering and biotechnology, enabling, for example the creation of bottom-up histoarchitectures of heterogeneous cells, or cell aggregates for in vitro high-throughput toxicological and therapeutic studies within 3D microenvironments. In this paper, a single-step process for creating peelable and resilient hydrogels, encapsulating arrays of biological cell aggregates formed by negative DEP has been devised. The dielectrophoretic trapping within low-energy regions of the DEP-dot array reduces cell exposure to high field stresses while creating distinguishable, evenly spaced arrays of aggregates. In addition to using an optimal combination of PEG diacrylate pre-polymer solution concentration and a novel UV exposure mechanism, total processing time was reduced. With a continuous phase medium of PEG diacrylate at 15% v/v concentration, effective dielectrophoretic cell patterned arrays and photo-polymerisation of the mixture was achieved within a 4 min period. This unique single-step process was achieved using a 30 s UV exposure time frame within a dedicated, wide exposure area DEP light box system. To demonstrate the developed process, aggregates of yeast, human leukemic (K562) and HeLa cells were immobilised in an array format within the hydrogel. Relative cell viability for both cells within the hydrogels, after maintaining them in appropriate iso-osmotic media, over a week period was greater than 90%.

Lessons in microcapsule assembly from imaging delivery of a bioluminescent enzyme.

Layer-by-layer assembled microcapsules have potential applications as delivery and biosensing systems, which make them attractive tools for use in various aspects of nanomedicine. We examined the effect of microcapsule location on activity of the bioluminescent enzyme luciferase in both intact capsules and following cell uptake. In intact capsules, the rate of reaction of luciferase was greatest for luciferase in the outer layer and least in the core. Following cell uptake, luciferase in the outer layer was rapidly reactive, and a similar rate of reaction and activity was observed for luciferase placed in capsule interior (core). By contrast, there was minimal activity detected when microcapsules with luciferase sandwiched between polyelectrolytes in a middle layer were delivered to cells. This study informs us of the availability of bioactive molecules located in different positions within microcapsules and will enable better microcapsule construction in line with the intended application, particularly delivery of functional proteins to cells.

Gene doping: gene delivery for olympic victory.

With one recently recommended gene therapy in Europe and a number of other gene therapy treatments now proving effective in clinical trials it is feasible that the same technologies will soon be adopted in the world of sport by unscrupulous athletes and their trainers in so called 'gene doping'. In this article an overview of the successful gene therapy clinical trials is provided and the potential targets for gene doping are highlighted. Depending on whether a doping gene product is secreted from the engineered cells or is retained locally to, or inside engineered cells will, to some extent, determine the likelihood of detection. It is clear that effective gene delivery technologies now exist and it is important that detection and prevention plans are in place.

The peritoneal tumour microenvironment of high-grade serous ovarian cancer.

High-grade serous ovarian cancer (HGSC) disseminates early and extensively throughout the peritoneal space, causing multiple lesions that are a major clinical problem. The aim of this study was to investigate the cellular composition of peritoneal tumour deposits in patient biopsies and their evolution in mouse models using immunohistochemistry, intravital microscopy, confocal microscopy, and 3D modelling. Tumour deposits from the omentum of HGSC patients contained a prominent leukocyte infiltrate of CD3(+) T cells and CD68(+) macrophages, with occasional neutrophils. Alpha-smooth muscle actin(+) (α-SMA(+) ) pericytes and/or fibroblasts surrounded these well-vascularized tumour deposits. Using the murine bowel mesentery as an accessible mouse peritoneal tissue that could be easily imaged, and two different transplantable models, we found multiple microscopic tumour deposits after i.p. injection of malignant cells. Attachment to the peritoneal surface was rapid (6-48 h) with an extensive CD45(+) leukocyte infiltrate visible by 48 h. This infiltrate persisted until end point and in the syngeneic murine ID8 model, it primarily consisted of CD3(+) T lymphocytes and CD68(+) macrophages with α-SMA(+) cells also involved from the earliest stages. A majority of tumour deposits developed above existing mesenteric blood vessels, but in avascular spaces new blood vessels tracked towards the tumour deposits by 2-3 weeks in the IGROV-1 xenografts and 6 weeks in the ID8 syngeneic model; a vigorous convoluted blood supply was established by end point. Inhibition of tumour cell cytokine production by stable expression of shRNA to CXCR4 in IGROV-1 cells did not influence the attachment of cells to the mesentery but delayed neovascularization and reduced tumour deposit size. We conclude that the multiple peritoneal tumour deposits found in HGSC patients can be modelled in the mouse. The techniques described here may be useful for assessing treatments that target the disseminated stage of this disease.

Preliminary investigations into the analgesic effects of topical ocular 1% morphine solution in dogs and cats.

OBJECTIVE: To perform preliminary evaluations into the ocular analgesic effect of topical 1% morphine in a clinical setting and to determine onset, duration and complications. STUDY DESIGN: Prospective, randomised, blinded clinical study. ANIMALS: Twenty six dogs and seventeen cats, all client-owned. METHODS: Dogs and cats with corneal ulceration requiring medical treatment or corneal conditions requiring surgery were included and randomly assigned to receive one drop of topical morphine (group M) or base solution (group B). Recordings were made prior to application and at 5, 10, 20, 30, 40, 50 and 60 minutes, then 2, 3, 4, 5 and 6 hours. Corneal aesthesiometry, blink rates and scores for blepharospasm (BLEPH), conjunctival hyperaemia (CH) and lacrimation (LAC) were recorded. Statistical analyses used anova, t-tests and Mann-Whitney U tests as relevant. RESULTS: No significant effect of treatment group on any recordings was found at any time point in either dogs or cats. Adverse effects of increased BLEPH, CH or blink rate were observed in six animals (three cats from group M and three dogs from group B), occurring within 5 minutes of drop application and lasting for between 10 minutes and 6 hours. CONCLUSIONS AND CLINICAL RELEVANCE: Topical ocular morphine showed no measurable analgesic effect against corneal pain in dogs and cats.