Expression of Potato virus Y cytoplasmic inclusion protein in tobacco results in disorganization of parenchyma cells, distortion of epidermal cells, and induces mitochondrial and chloroplast abnormalities, formation of membrane whorls and atypical lipid accumulation

Infection of plant cells by potyviruses induces the formation of cytoplasmic inclusions ranging in size from 200 to 1000 nm. To determine if the ability to form these ordered, insoluble structures is intrinsic to the potyviral cytoplasmic inclusion protein, we have expressed the cytoplasmic inclusion protein from Potato virus Y in tobacco under the control of the chrysanthemum ribulose-1,5-bisphosphate carboxylase small subunit promoter, a highly active, green tissue promoter. No cytoplasmic inclusions were observed in the leaves of transgenic tobacco using transmission electron microscopy, despite being able to clearly visualize these inclusions in Potato virus Y infected tobacco leaves under the same conditions. However, we did observe a wide range of tissue and sub-cellular abnormalities associated with the expression of the Potato virus Y cytoplasmic inclusion protein. These changes included the disruption of normal cell morphology and organization in leaves, mitochondrial and chloroplast internal reorganization, and the formation of atypical lipid accumulations. Despite these significant structural changes, however, transgenic tobacco plants were viable and the results are discussed in the context of potyviral cytoplasmic inclusion protein function.

Cyclic strain disrupts endothelial network formation on Matrigel

Most forms of tissue healing depend critically on revascularisation. In soft tissues and in vitro, mechanical stimuli have been shown to promote vessel-forming activity. However, in bone defects, increased interfragmentary motion impairs vascular regeneration. Because these effects seem contradictory, we aimed to determine whether a range of mechanical stimuli exists in which angiogenesis is favoured. A series of cyclic strain magnitudes were applied to a Matrigel-based “tube formation” assay and the total lengths of networks formed by human microvascular endothelial cells measured at 24 h. Network lengths were reduced at all strain levels, compared to unstretched controls. However, the levels of pro-angiogenic matrix metalloproteases-2 and -9 in the corresponding conditioned media were unchanged by strain, and vascular endothelial growth factor was uniformly elevated in stretched conditions. By repeating the assay with the addition of conditioned media from mesenchymal stem cells cultivated in similar conditions, paracrine stimuli were shown to increase network lengths, but not to alter the negative effect of cyclic stretching. Together, these results demonstrate that directly applied periodic strains can inhibit endothelial organisation in vitro, and suggest that this may be due to physical disruption rather than biochemical modulation. Most importantly, the results indicate that the straining of endothelial cells and their assembly into vascular-like structures must be studied simultaneously to adequately characterise the mechanical influence on vessel formation.

Processing of polycaprolactone and polycaprolactone-based copolymers into 3D scaffolds and their cellular responses

Synthetic polymers have attracted much attention in tissue engineering due to their ability to modulate biomechanical properties. This study investigated the feasibility of processing poly(varepsilon-caprolactone) (PCL) homopolymer, PCL-poly(ethylene glycol) (PEG) diblock, and PCL-PEG-PCL triblock copolymers into three-dimensional porous scaffolds. Properties of the various polymers were investigated by dynamic thermal analysis. The scaffolds were manufactured using the desktop robot-based rapid prototyping technique. Gross morphology and internal three-dimensional structure of scaffolds were identified by scanning electron microscopy and micro-computed tomography, which showed excellent fusion at the filament junctions, high uniformity, and complete interconnectivity of pore networks. The influences of process parameters on scaffolds' morphological and mechanical characteristics were studied. Data confirmed that the process parameters directly influenced the pore size, porosity, and, consequently, the mechanical properties of the scaffolds. The in vitro cell culture study was performed to investigate the influence of polymer nature and scaffold architecture on the adhesion of the cells onto the scaffolds using rabbit smooth muscle cells. Light, scanning electron, and confocal laser microscopy showed cell adhesion, proliferation, and extracellular matrix formation on the surface as well as inside the structure of both scaffold groups. The completely interconnected and highly regular honeycomb-like pore morphology supported bridging of the pores via cell-to-cell contact as well as production of extracellular matrix at later time points. The results indicated that the incorporation of hydrophilic PEG into hydrophobic PCL enhanced the overall hydrophilicity and cell culture performance of PCL-PEG copolymer. However, the scaffold architecture did not significantly influence the cell culture performance in this study.

Historical and Personal Perspectives on the Fall of the Berlin Wall, 1989: address to 20th anniversary forums.

The story of the fall of the Berlin Wall was an aspect of the “imagination gap” that we had to wrestle with as journalists covering the collapse of the Eastern Bloc in Europe. It was scarcely possible to believe what you found yourself reporting, and that work became a two-track process. On one hand a mass social movement was dictating the pace and direction of events; on the other, the institutional business of politics as usual, to provide a framework for all the change that was happening, had to be managed – and reported on. In later analyseds we could see, that crisis in the Soviet Union led to the crisis over the Berlin Wall; and from the fall of the Wall, came Germany’s reunification, and with that also, formation of the European Union as it is today. The government of the Federal Republic of Germany convinced its neighbours that a reunited Germany, within an expanded EU, would be a very acceptable “European Germany” -- not the leader of a “German Europe”. It committed itself financially, supporting the new Euro currency. The former communist states of Eastern Europe demanded to join and expand the EU; in order to remove themselves from the Soviet Union, enjoy human rights, and share in Western prosperity. So today, following on from the events of 1989, the European Union is an amalgam of 27 member countries, with close to 500 million citizens and accounting for 30 % of world Gross National Product.

Patterns of fitting cosmetically tinted contact lenses

Cosmetically tinted soft contact lenses are an attractive option for contact lens wearers. Data that we have gathered from annual contact lens fitting surveys demonstrate that those wearing tinted lenses are more likely to be female (4.6% of all soft lenses fitted vs. 1.6% for males; p < 0.0001) and younger (27 11 years vs. 33 13 years for those wearing non-tinted lenses; p < 0.0001). Tinted lenses tend to be worn more on a part-time basis and are replaced less frequently than non-tinted lenses. The decline in fitting tinted lenses over the past 12 years may be due to (a) the current limited availability of tinted lenses in silicone hydrogel materials and daily disposable replacement frequencies, which together represent a significant majority (78%) of new soft lenses fits today, (b) growing concerns among lens wearers and practitioners relating to the risks of complications associated with the wearing of tinted lenses, and (c) reduced promotion of such lenses by the contact lens industry.

Factors influencing the prescribing of hydrogel contact lenses

Despite the current market popularity of silicone hydrogel contact lenses, conventional hydrogel lenses still represent a significant proportion of the overall contact lens market. Data gathered from annual UK contact lens fitting surveys over the past 13 years indicate that 64% of hydrogel lenses prescribed during this period were of low/medium water content (<60% water). Whereas, in the past, practitioners would choose the water content of a hydrogel lens to meet specific clinical needs, this choice appears today to be largely governed by product availability.

Enhancement of DNA repair using topical T4 endonuclease V does not inhibit melanoma formation in Cdk4R24C/R24C/Tyr-NrasQ61K mice following neonatal UVR

To further investigate the use of DNA repair-enhancing agents for skin cancer prevention, we treated Cdk4R24C/R24C/NrasQ61K mice topically with the T4 endonuclease V DNA repair enzyme (known as Dimericine) immediately prior to neonatal ultraviolet radiation (UVR) exposure, which has a powerful effect in exacerbating melanoma development in the mouse model. Dimericine has been shown to reduce the incidence of basal-cell and squamous cell carcinoma. Unexpectedly, we saw no difference in penetrance or age of onset of melanoma after neonatal UVR between Dimericine-treated and control animals, although the drug reduced DNA damage and cellular proliferation in the skin. Interestingly, epidermal melanocytes removed cyclobutane pyrimidine dimers (CPDs) more efficiently than surrounding keratinocytes. Our study indicates that neonatal UVR-initiated melanomas may be driven by mechanisms other than solely that of a large CPD load and/or their inefficient repair. This is further suggestive of different mechanisms by which UVR may enhance the transformation of keratinocytes and melanocytes.

A population survey of the penetrance of contact lens wear in Australia : rationale, methodology and results

Purpose: A population based, cross-sectional telephone survey was conducted to estimate the total penetrance of contact lens wear in Australia. Methods: A total of 42,749 households around Australia were randomly selected from the national electronic telephone directory based on postcode distribution. Before contact was attempted, letters of introduction were sent. The number of individuals and contact lens wearers in each household was ascertained and lens wearers were interviewed to determine details of lens type and mode of wear using a structured questionnaire. Results: Of households contacted, 59.2% (19,171/32,405) agreed to participate. Response rates were only marginally higher amongst households that first received a letter of introduction. In these households, 35,914 individuals were identified, of which, 1,798 were contact lens wearers. The penetrance of contact lens wear during the study period was 5.01% (95% CI: 4.78-5.24). Soft hydrogel lenses had the largest penetrance in the community, (66.7% of all wearers), however, their market share decreased significantly over the study period with increased uptake of newly introduced lens types. Conclusions: The penetrance of contact lens wear concurs with market estimates and equates to approximately 680,000 contact lens wearers aged between 15 and 64 years in Australia. The low response rate obtained in this study highlights the difficulty in contemporary use of telephone survey methodology

Kinematics of intracellular chlamydiae provide evidence for contact-dependent development

A crucial process of chlamydial development involves differentiation of the replicative reticulate body (RB) into the infectious elementary body (EB). We present experimental evidence to provide support for a contact-dependent hypothesis for explaining the trigger involved in differentiation. We recorded live-imaging of Chlamydia trachomatis-infected McCoy cells at key times during development and tracked the temporospatial trajectories of individual chlamydial particles. We found that movement of the particles is related to development. Early to mid-developmental stages involved slight wobbling of RBs. The average speed of particles increased sharply at 24 h postinfection (after the estimated onset of RB to EB differentiation). We also investigated a penicillin-supplemented culture containing EBs, RBs, and aberrantly enlarged, stressed chlamydiae. Near-immobile enlarged particles are consistent with their continued tethering to the chlamydial inclusion membrane (CIM). We found a significantly negative, nonlinear association between speed and size/type of particles, providing further support for the hypothesis that particles become untethered near the onset of RB to EB differentiation. This study establishes the relationship between the motion properties of the chlamydiae and developmental stages, whereby wobbling RBs gradually lose contact with the CIM, and RB detachment from the CIM is coincidental with the onset of late differentiation.

Tear film surface quality with soft contact lenses using dynamic-area high-speed videokeratoscopy

Objectives. To evaluate the performance of the dynamic-area high-speed videokeratoscopy technique in the assessment of tear film surface quality with and without the presence of soft contact lenses on eye. Methods. Retrospective data from a tear film study using basic high-speed videokeratoscopy, captured at 25 frames per second, (Kopf et al., 2008, J Optom) were used. Eleven subjects had tear film analysis conducted in the morning, midday and evening on the first and seventh day of one week of no lens wear. Five of the eleven subjects then completed an extra week of hydrogel lens wear followed by a week of silicone hydrogel lens wear. Analysis was performed on a 6 second period of the inter-blink recording. The dynamic-area high-speed videokeratoscopy technique uses the maximum available area of Placido ring pattern reflected from the tear interface and eliminates regions of disturbance due to shadows from the eyelashes. A value of tear film surface quality was derived using image rocessing techniques, based on the quality of the reflected ring pattern orientation. Results. The group mean tear film surface quality and the standard deviations for each of the conditions (bare eye, hydrogel lens, and silicone hydrogel lens) showed a much lower coefficient of variation than previous methods (average reduction of about 92%). Bare eye measurements from the right and left eyes of eleven individuals showed high correlation values (Pearson’s correlation r = 0.73, p < 0.05). Repeated measures ANOVA across the 6 second period of measurement in the normal inter-blink period for the bare eye condition showed no statistically significant changes. However, across the 6 second inter-blink period with both contact lenses, statistically significant changes were observed (p < 0.001) for both types of contact lens material. Overall, wearing hydrogel and silicone hydrogel lenses caused the tear film surface quality to worsen compared with the bare eye condition (repeated measures ANOVA, p < 0.0001 for both hydrogel and silicone hydrogel). Conclusions. The results suggest that the dynamic-area method of high-speed videokeratoscopy was able to distinguish and quantify the subtle, but systematic worsening of tear film surface quality in the inter-blink interval in contact lens wear. It was also able to clearly show a difference between bare eye and contact lens wearing conditions.

Macroentrepreneurship : an expanded contact for entrepreneurship

The purpose if this paper is to show how entrepreneurial practices can be applied at macro level by industry and government bodies, particularly to identify opportunity areas in target industries and thereby promote new business ventures that best serve the economy and society. A new macro context is argued for entrepreneurial activity based on marketing and entrepreneurship principles, and their specialization areas or social and societal marketing, macromarketing and social entrepreneurship. An example of government and industry involvement in industry opportunity identification is outlines to demonstrate macro level entrepreneurial activity, showing how, in the same way that macromarketing seeks to address the bigger issues and the links between marketing systems and society, so too can entrepreneurship use this perspective to achieve its aims and contribute more effectively to the betterment of society. This paper makes and original contribution by demonstrating a new, expanded context for entrepreneurship's scholarly domain and its practice, showing how its key concepts can be effectively applied at industry level to provide a catalyst for development.

Eyelid pressure and contact with the ocular surface

Purpose To investigate static upper eyelid pressure and contact with the ocular surface in a group of young adult subjects. Methods Static upper eyelid pressure was measured for 11 subjects using a piezoresistive pressure sensor attached to a rigid contact lens. Measures of eyelid pressure were derived from an active pressure cell (1.14 mm square) beneath the central upper eyelid margin. To investigate the contact region between the upper eyelid and ocular surface, we used pressure sensitive paper and the lissamine-green staining of Marx’s line. These measures combined with the pressure sensor readings were used to derive estimates of eyelid pressure. Results The mean contact width between the eyelids and ocular surface estimated using pressure sensitive paper was 0.60 ± 0.16 mm, while the mean width of Marx’s line was 0.09 ± 0.02 mm. The mean central upper eyelid pressure was calculated to be 3.8 ± 0.7 mmHg (assuming that the whole pressure cell was loaded), 8.0 ± 3.4 mmHg (derived using the pressure sensitive paper imprint widths) and 55 ± 26 mmHg (based on contact widths equivalent to Marx’s line). Conclusions The pressure sensitive paper measurements suggest that a band of the eyelid margin, significantly larger than the anatomical zone of the eyelid margin known as Marx’s line, has primary contact with the ocular surface. Using these measurements as the contact between the eyelid margin and ocular surface, we believe that the mean pressure of 8.0 ± 3.4 mmHg is the most reliable estimate of static upper eyelid pressure.

Signs, symptoms and comorbidities in contact lens-related microbial keratitis

Purpose: The aim of this study was to characterize the clinical signs, symptoms, and ocular and systemic comorbidities in a large case series of contact lens-related microbial keratitis. Methods: Two hundred ninety-seven cases of contact lens-related microbial keratitis, aged between 15 and 64 years were detected through surveillance of hospital and community based ophthalmic practitioners in Australia and New Zealand. Full clinical data were available for 190 cases and 90 were interviewed by telephone. Clinical data included the size, location, and degree of anterior chamber response. Symptom data were available from the practitioner and from participant self-report. Associations between symptoms and disease severity were evaluated. Data on ocular and systemic disease were collected from participants and practitioners. The frequency of comorbidities was compared between the different severities of disease and to population norms. Results: More severe disease was associated with greater symptom severity and pain was the most prevalent symptom reported. Ninety-one percent of cases showed progression of ocular symptoms after lens removal, and symptom progression was associated with all severities of disease. Twenty-five percent of cases reported prior episodes requiring emergency attention. Thyroid disease (p 0.05) and self-reported poor health (p 0.001) were more common in cases compared with age-matched population norms. Discussion: Information on the signs, symptoms, and comorbidities associated with contact lens-related microbial keratitis may be useful in patient education and for practitioners involved in the fitting of lenses and management of complications. Although pain was the most common symptom experienced, progression of symptoms despite lens removal was close to universal. Poor general health, particularly respiratory disease and thyroid disease was more common in cases than in the general population, which may prompt practitioners to recommend flexibility in wear schedules when in poor health or the selection of a lower risk wear schedule in at risk patients

Practitioner influence on contact lens prescribing in the UK

Contact lenses are mainly fitted by registered optometrists and contact lens opticians in the UK. Data we have gathered from annual contact lens fitting surveys over the past 12 years indicate that, on average, registered optometrists and contact lens opticians undertake 3.2 and 7.1 contact lens fits per week (p < 0.0001). More experienced practitioners tend to fit older patients. Practitioners fitting more lenses per year tend to fit a higher proportion of soft lenses. Contact lens opticians tend to fit a higher proportion of patients with planned replacement and daily disposable lenses compared with optometrists.