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Background/aim: Nestin is an intermediate filament marker for neural progenitor cells. The authors aimed to identify nestin positive cells in adult human retina and within surgically removed epiretinal membranes. Methods: [...]

Models of invasive species spread often assume that landscapes are spatially homogeneous; thus simplifying analysis but potentially reducing accuracy. We extend a recently developed partial differential equation model for invasive conifer spread to account for spatial heterogeneity in parameter values and introduce a method to obtain key outputs (e.g. spread rates) from computational simulations. Simulations produce patterns of spatial spread which appear qualitatively similar to observed patterns in grassland ecosystems invaded by exotic conifers, validating our spatially explicit strategy. We find that incorporating spatial variation in different parameters does not significantly affect the evolution of invasions (which are characterised by a long quiescent period followed by rapid evolution towards to a constant rate of invasion) but that distributional assumptions can have a significant impact on the spread rate of invasions. Our work demonstrates that spatial variation in site-suitability or other parameters can have a significant impact on invasions and must be considered when designing models of invasive species spread., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Hughes et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Purpose: To demonstrate MR T 1 mapping in vivo as a method to non-invasively estimate vitreous oxygen concentration in ischemic eye disease., Methods: Patients with ischemic eye disease (central retinal vein occlusion, ocular ischemic syndrome, and proliferative diabetic retinopathy) were prospectively recruited. MRI was performed on each patient before any treatment, with T 1 mapping acquired using an inversion recovery TrueFISP sequence at several inversion times, from a single slice positioned through the center of both eyes in the axial oblique plane. A phantom study measuring seven different concentrations of vitronectin, a protein released in ischemic eye disease, was undertaken to determine its potential confounding effect on T 1 ., Results: Ten participants were recruited (eight central retinal vein occlusion, one ocular ischemic syndrome, and one proliferative diabetic retinopathy). Of the eight central retinal vein occlusion cases, there was a statistically different vitreous T 1 in the diseased eye compared to the healthy control eye (4.306 vs. 4.518 s, p = 0.008). T 1 times did not significantly alter across the range of vitronectin concentrations., Conclusions: Ischemic eye disease decreases vitreous T 1 , potentially implying an increase in vitreous partial pressure of oxygen (pO 2 ) concentration given what is known from the relationship between 1/T 1 and pO 2 . Potential theories for this unexpected result are discussed. This study provides further data on this technique, with potential clinical application in eye disease., (© 2023 International Society for Magnetic Resonance in Medicine.)

Background: Incidence of differentiated thyroid cancer has increased in the United States and globally with disproportionate increases observed among women. Recent data suggest that factors other than increased detection may underlie this increase. To understand incidence and survival patterns in differentiated thyroid cancer during a time period of increasing imaging, we examined data from a contemporary population-based sample of U.S. reproductive-aged women. Methods: Women aged 20-49 years ( N  = 61,552) diagnosed with papillary thyroid cancer (PTC) or follicular thyroid cancer (FTC) during 2000-2016 were identified from the U.S. National Cancer Institute Surveillance, Epidemiology, and End Results 18 registries database. For each age decade (20-29, 30-39, 40-49 years), we estimated age-adjusted average annual percentage changes in incidence using segmented and unsegmented regression models and 15-year survival. Results were stratified by race/ethnicity and cancer stage. Results: The estimated incidence of PTC increased during 2000-2016 among women aged 20-29 years and during 2000-2012 among women aged 30-49 years. During 2012-2016, incidence stabilized among women aged 30-39 years and decreased among women aged 40-49 years. For FTC, incidence decreased slightly among women aged 20-29 years and was rather stable among those aged 30-49 years during 2000-2016, although increases were observed among non-Hispanic black women aged 30-49 years. By stage, the percentage increase in PTC incidence was largest for regional disease. Fifteen-year estimated survival was generally high but somewhat lower among women aged 40-49 years than those aged 20-39 years. Survival was similar for PTC and FTC except among women aged 20-29 years, for whom survival was modestly lower with FTC than PTC. Conclusions: Our findings confirm increasing incidence of PTC among U.S. women aged 20-29 years, a recent stabilization of PTC incidence in women 30-49 years, and stable to decreasing incidence of FTC. Increased detection based on imaging is unlikely to fully explain the continued increase in PTC incidence, given the increasing incidence of regional disease and routine imaging occurring less often among premenopausal than postmenopausal women. Although survival is generally high, treatment often requires surgery and lifelong medications. Further investigations into contributors to these trends are warranted to reduce future morbidity in reproductive-aged women.

Mammalian cells were grown to high density in a 3,000 L culture using perfusion with hollow fibers operated in a tangential flow filtration mode. The high-density culture was used to inoculate the production stage of a biomanufacturing process. At constant permeate flux operation, increased transmembrane pressures (TMPs) were observed on the final day of the manufacturing batches. Small scale studies suggested that the filters were not irreversibly fouled, but rather exposed to membrane concentration polarization that could be relieved by tangential sweeping of the hollow fibers. Studies were undertaken to analyze parameters that influence the hydrodynamic profile within hollow fibers; including filter area, cell density, recirculation flow rate, and permeate flow rate. Results indicated that permeate flow rate had the greatest influence on modulating TMP. Further evaluation showed a significant decrease in TMP when permeate flow was reduced, and this occurred without any negative effect on cell growth or viability. Hence, a 30% reduction of permeate flow rate was implemented at manufacturing scale. A stable operation was achieved as TMP was successfully reduced by 75% while preserving all critical factors for performance in the perfusion bioreactor., (© 2020 American Institute of Chemical Engineers.)

Over the past decade, abnormalities have been documented in white-tailed deer (Odocoileus virginianus) in west-central Montana. Hypotheses proposed to explain these anomalies included contact with endocrine disrupting pesticides, such as imidacloprid. We evaluated the effects of imidacloprid experimentally at the South Dakota State University Wildlife and Fisheries Captive Facility where adult white-tailed deer females and their fawns were administered aqueous imidacloprid (an untreated control, 1,500 ng/L, 3,000 ng/L, and 15,000 ng/L). Water consumption, thyroid hormone function, behavioral responses, and skull and jawbone measurements were compared among treatments. Additionally, liver, spleen, genital, and brain imidacloprid concentrations were determined by an enzyme-linked immunosorbent assay (ELISA). Results indicated that 1) control deer consumed more water than treatment groups, 2) imidacloprid was present in the organs of our control group, indicating environmental contamination, 3) as imidacloprid increased in the spleen, fawn survival, thyroxine levels, jawbone lengths, body weight, and organ weights decreased, 4) adult female imidacloprid levels in the genitals were negatively correlated with genital organ weight and, 5) behavioral observations indicated that imidacloprid levels in spleens were negatively correlated with activity levels in adult females and fawns. Results demonstrate that imidacloprid has direct effects on white-tailed deer when administered at field-relevant doses.

The ability to control charge heterogeneity in monoclonal antibodies is important to demonstrate product quality comparability and consistency. This article addresses the control of C-terminal lysine processing through copper supplementation to yeast hydrolysate powder, a raw material used in the cell culture process. Large-scale production of a murine cell line exhibited variation in the C-terminal lysine levels of the monoclonal antibody. Analysis of process data showed that this variation correlated well with shifts in cell lactate metabolism and pH levels of the production culture. Small-scale studies demonstrated sensitivity of the cells to copper, where a single low dose of copper to the culture impacted cell lactate metabolism and C-terminal lysine processing. Subsequent analytical tests indicated that the yeast hydrolysate powder, added to the basal media and nutrient feed in the process, contained varying levels of trace copper across lots. The measured copper concentrations in yeast hydrolysate lots correlated well with the variation in lactate and pH trends and C-terminal lysine levels of the batches in manufacturing. Small-scale studies further demonstrated that copper supplementation to yeast hydrolysate lots with low concentrations of copper can shift the metabolic performance and C-terminal lysine levels of these cultures to match the control, high copper cultures. Hence, a strategy of monitoring, and if necessary supplementing, copper in yeast-hydrolysate powders resulted in the ability to control and ensure product quality consistency. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:463-468, 2017., (© 2017 American Institute of Chemical Engineers.)

All individuals are a part of at least one culture. These cultural contingencies shape behavior, behavior that may or may not be acceptable or familiar to behavior analysts from another culture. To better serve individuals, assessments and interventions should be selected with a consideration of cultural factors, including cultural preferences and norms. The purpose of this paper is to provide suggestions to serve as a starting point for developing behavior analysts' cultural awareness skills. We present strategies for understanding behavior analysts' personal cultural values and contingencies and those of their clients, integrating cultural awareness practices into service delivery, supervision, and professional development, and becoming culturally aware in everyday practice.

Purpose: The purpose of this study was to monitor a case of acute exudative polymorphous vitelliform maculopathy detected following trauma., Methods: Clinical examination included fundus photographs, optical coherence tomography (OCT) and fluorescein angiography. Pattern and full-field electroretinograms (PERG; ERG) and serial electrooculograms (EOG) were performed, incorporating the international standards., Results: A 45-year-old Caucasian woman developed blurring of vision in both eyes 5 days after a serious road traffic accident. On examination, bilateral serous macular detachments and multiple small yellow subretinal lesions were observed in both eyes associated with areas of serous retinal detachment superior and inferior to the yellow lesions in the left eye. OCT showed retinal elevation with a band of high reflectance on the outer retina but no intra-retinal fluid or elevation of the retinal pigment epithelium (RPE). There was bilateral EOG reduction in keeping with generalised RPE dysfunction and pattern ERG evidence of left macular involvement. After 9 months, the patient reported spontaneous improvement in vision with gravitational settling and coalescence of the subretinal yellow deposits. At 37 months, there was improvement in the EOG and visual acuity., Conclusions: A rare case of AEPVM is described that was detected following trauma and which gradually improved over 37 months. The EOG showed evidence of generalised RPE dysfunction that resolved and may be useful in the monitoring of AEPVM.

Purpose: To study the effect of pars plana vitrectomy (PPV) on vitreous oxygenation (pO2) using magnetic resonance imaging (MRI)., Methods: Patients due to undergo PPV for either macular hole or epiretinal membrane were recruited. MRI scanning was performed 1 week before and at least 3 months after PPV. MRI T1 mapping was performed using an inversion recovery-true fast imaging with steady-state precession (TrueFISP) sequence at several inversion times, from a single slice positioned through the center of both eyes in the axial oblique plane. Additional phantom data were measured in porcine vitreous, to define the relationships between T1 relaxation times and balanced salt solution (BSS), to simulate human vitreous and aqueous, respectively, for a suitable pO2 range (5-70 mm Hg). Pre-PPV pO2 was also measured intraoperatively using a polarographic oxygen probe., Results: Eleven participants (age range 59-84) were recruited; two declined the post-PPV scan. Corrected T1 times indicated that the mean (±SD) pO2 increased significantly following PPV, from 13.2 ± 5.8 to 34.5 ± 8.0 mm Hg (P < 0.001). In the nonsurgical (control) eye, pO2 did not change significantly from the first to second MRI scan (13.7 ± 7.8 vs. 16.3 ± 8.7 mm Hg, P = 0.239). Mean pO2 measured intraoperatively was 7.2 ± 0.6 mm Hg (n = 10)., Conclusions: These results confirm that vitrectomy substantially increases vitreous pO2. MRI is a noninvasive technique that can be used to study vitreous oxygenation in both vitrectomized and nonvitrectomized eyes.

Purpose: To report a case of visual loss immediately after hand, foot, and mouth disease and demonstrate the high-resolution optical coherence tomography findings., Methods: A retrospective case report of a 19-year-old nursery worker with resolving hand, foot, and mouth disease and acute unilateral visual loss., Results: The clinical features were characteristic of unilateral acute idiopathic maculopathy. High-resolution optical coherence tomography demonstrated highly reflective subretinal material at the macula of one eye with disruption of the photoreceptor inner segment/outer segment junction. Vision remained poor for 4 weeks when there was rapid recovery coinciding with reconstitution of the inner segment/outer segment junction on optical coherence tomography., Conclusion: Unilateral acute idiopathic maculopathy may be caused by Coxsackievirus infection. Optical coherence tomography and clinical findings suggest an acute viral retinal pigment epitheliitis to be the main pathologic feature.

Purpose: The purpose of this study was to report the case of a patient with acute retinal necrosis who developed neurotoxicity while being treated with oral valaciclovir antiviral therapy., Methods: Testing used was clinical examination including fundus photographs, renal biochemistry, cerebrospinal fluid analysis, and renal ultrasound., Results: A 63-year-old, healthy, white woman was being treated with oral valaciclovir for acute retinal necrosis. Several days later, she developed visual hallucinations and confusion. Renal biochemistry was abnormal, and renal ultrasound was normal. Cerebrospinal fluid cellularity was normal, but cerebrospinal fluid 9-carboxymethoxymethylguanine, an aciclovir metabolite, level was elevated. Treatment was changed to intravenous aciclovir titrated based on serum aciclovir levels with intravenous fluid supplementation to good effect., Conclusion: Regular monitoring of renal function and ample fluid intake are important when high doses of aciclovir/valaciclovir are used, even in patients with no previous history of renal failure. Monitoring of serum aciclovir level is useful in those with renal impairment to titrate doses, and high-dose oral valaciclovir should be used with caution.

The terpenoid indole alkaloid (TIA) pathway in Catharanthus roseus produces two important anticancer drugs, vinblastine and vincristine, in very low yields. This study focuses on overexpressing several key genes in the upper part of the TIA pathway in order to increase flux toward downstream metabolites within hairy root cultures. Specifically, we constructed hairy root lines with inducible overexpression of 1-deoxy-D-xylulose synthase (DXS) or geraniol-10-hydroxylase (G10H). We also constructed hairy root lines with inducible expression of DXS and anthranilate synthase α subunit (ASA) or DXS and G10H. DXS overexpression resulted in a significant increase in ajmalicine by 67%, serpentine by 26% and lochnericine by 49% and a significant decrease in tabersonine by 66% and hörhammericine by 54%. Co-overexpression of DXS and G10H caused a significant increase in ajmalicine by 16%, lochnericine by 31% and tabersonine by 13%. Likewise, DXS and ASA overexpression displayed a significant increase in hörhammericine by 30%, lochnericine by 27% and tabersonine by 34%. These results point to the need for overexpressing multiple genes within the pathway to increase the flux toward vinblastine and vincristine., (Copyright © 2010 Elsevier Inc. All rights reserved.)

Background: The incidence of new infection with syphilis is increasing, particularly in men who have sex with men, with HIV co-infection common. There has been a corresponding increase in ophthalmic manifestations that can be varied in presentation., Methods: Thirteen consecutive patients with syphilitic uveitis presenting to two ophthalmic departments in Sydney are described., Results: Twelve patients were male, of whom 10 were homosexual and six HIV-positive. Peripheral retinitis with panuveitis was the commonest ophthalmic presentation (n = 7, 54%), and six cases were initially treated with vitreous tap and intravitreal foscarnet as a precaution in case of viral retinitis. Retinitis was present in six of six (100%) HIV-positive and only one of seven (14%) HIV-negative patients (χ² 10.6, P < 0.01). Other ophthalmic presentations included anterior uveitis, vitritis, multifocal choroiditis, scleritis and papillitis. All patients responded to 10-14 days' intravenous penicillin with good final visual outcomes (6/12 or better in all eyes)., Conclusions: This case series reinforces the importance of considering syphilis in the differential diagnosis of many ocular presentations, but in particular retinitis. Retinitis appears to be the predominant presentation in HIV-infected individuals, suggesting that HIV infection may somehow modulate the disease., (© 2010 The Authors. Clinical and Experimental Ophthalmology © 2010 Royal Australian and New Zealand College of Ophthalmologists.)

Purpose: To compare corneal endothelial cell loss after cataract surgery by phacoemulsification and by Aqualase., Methods: This was a prospective, randomized study of 75 eyes of 75 patients undergoing cataract surgery. Patients were randomly assigned to receive either phacoemulsification or Aqualase for cataract removal. Specular microscopy was used to calculate endothelial cell counts preoperatively and 3 weeks and 6 months afterwards. Best-corrected visual acuity was also measured. The t-test was used to detect statistical significance., Results: The mean endothelial cell loss was 8.1% in the phacoemulsification group and 6.8% in the Aqualase group. There was no statistically significant difference in the amount of endothelial cell loss or in visual outcome between the 2 groups. The number of Aqualase pulses tended to increase with nuclear density while the phacoemulsification time showed little variation with nuclear subtype., Conclusion: Endothelial cell loss after cataract surgery is similar whether phacoemulsification or Aqualase is used. Aqualase can be considered to be as safe as phacoemulsification with regard to corneal trauma and is a useful alternative especially for soft cataracts.

Jasmonic acid (JA) activates the transcriptional regulator ORCA3, which has a role in regulating the terpenoid indole alkaloid (TIA) pathway within Catharanthus roseus. The TIA pathway leads to the production of the anticancer drugs vinblastine and vincristine. This work explores the transient effects of overexpressing ORCA3 under the control of a glucocorticoid-inducible promoter system in C. roseus hairy roots along with the simultaneous feeding of JA. The changes in TIA metabolites and in mRNA transcripts of pathway genes and regulators were tracked for 72h. Upon induction of ORCA3 expression and elicitation with JA, ORCA3 transcripts increased 170-fold whereas ORCA3 expression caused an 89-fold increase and JA elicitation caused a 5-fold increase in ORCA3 transcripts. JA treatment caused the largest increase in TIA metabolites and transcripts of pathway genes. These transcripts displayed a transient response with the maximum expression reached between 12 and 24h. In the samples overexpressing ORCA3, the largest increase in the transcripts of ZCT1 and ZCT2 (ZCT-zinc finger-binding protein), TIA transcriptional repressors, coincided with the largest increase in ORCA3 transcripts. This counter response of transcriptional repressors may explain why the large increase in ORCA3 transcripts do not correspond with larger increases in transcripts of TIA pathway genes.

Aims: To describe the clinical and radiological features of orbital apex syndromes caused by presumed tuberculosis (TB)., Methods: A review of the discharge summaries of all patients seen in the Medical Eye Unit (MEU), St Thomas' Hospital between 1975 and 2006 identified seven patients with a diagnosis of orbital apex syndrome or optic neuropathy attributable to TB. Case notes and radiology were reviewed retrospectively for each patient., Results: All of the patients were born outside the UK and were HIV-negative. Four presented during 2005-6. Six of the seven patients presented with a visual acuity (VA) of less than counting fingers (CF), but all achieved a VA of 6/9 or better after a median 3 weeks of treatment with antituberculous treatment and systemic corticosteroids. All patients had constitutional symptoms. Chest x ray and CSF were normal in each case, but MRI was abnormal in six., Conclusions: We report a recent cluster of tuberculous orbital apex syndromes, in the context of an increasing incidence of TB in the UK. The condition is difficult to diagnose, but the combination of high-dose corticosteroids and anti-TB medication was rapidly effective and achieved a good outcome in all cases.

Purpose: To describe the vitreoretinal complications in a cohort of patients with osteoodontokeratoprosthesis (OOKP) and discuss surgical management., Methods: Review of notes of 35 OOKP cases performed at the Sussex Eye Hospital (Brighton, United Kingdom) between January 1999 and December 2005 was performed., Results: The overall incidence of vitreoretinal complications was 22.8%, which included vitreous hemorrhage (3 patients), rhegmatogenous retinal detachment (3 patients), endophthalmitis with retinal detachment complicating lamina resorption and optic extrusion (2 patients), and intraoperative choroidal hemorrhage (1 patient). Preexisting aphakia was associated with rhegmatogenous retinal detachment (P < 0.05, chi2 = 4.36). Five patients required pars plana vitrectomy, which was performed either endoscopically (two cases) or using a binocular indirect viewing system (three cases) with one case requiring removal of the OOKP and insertion of a temporary keratoprosthesis. Retinal detachment repair was attempted on four of five patients but was successful for only one. Vitreous hemorrhage without retinal detachment required vitrectomy in one case, while two cases cleared spontaneously., Conclusions: Eyes receiving OOKP are prone to vitreoretinal complications, with retinal detachment associated with a poor prognosis. Thicker OOKP laminae and lamina bulk screening will hopefully reduce the risk of endophthalmitis due to unexpected resorption.

Introduction: The inadvertent intra-ocular administration of benzalkonium chloride-preserved hydroxypropyl methylcellulose during cataract surgery at another hospital in 1999 resulted in toxic corneal endothelial injury and profound postoperative corneal oedema as a result of endothelial decompensation. The long-term effect of this adverse event was assessed., Methods: All 19 patients were invited to return for examination including corneal endothelial specular microscopy and pachymetry seven years after the incident. Results were compared with data from one year after the incident., Results: Five patients attended for examination, one had received a penetrating keratoplasty and was, therefore, excluded. Ten patients had died and four had moved out of the region and were unable to attend. All four study patients were pain free and achieved 6/12 or better. Mean central corneal thickness reduced by 13% from 652.6 microm at one year to 563.4 microm. Mean central corneal endothelial cell density (n = 3) increased 28% from 663.7 cells/mm(2) at one year to 835.7 cells/mm(2) (p<0.05)., Conclusions: After toxic injury, corneal endothelial function may have a remarkable capacity for recovery even after the first postoperative year. The rise in central endothelial cell density may represent cell migration from less affected areas or cellular proliferation. Should this unfortunate event recur, clinicians may expect continued recovery beyond one year.

We present a case of complete blindness following severe dengue haemorrhagic fever complicated by anaemia and a dialysis-related episode of profound hypotension. The clinical and radiological features indicated an optic neuropathy, most likely ischaemic in aetiology. The features of posterior ischaemic optic neuropathy and differential diagnosis are discussed.

Purpose: To report our experience of using Aqualase technology for cataract extraction., Methods: In total, 33 patients (20 females; mean age 71.4 years) underwent cataract surgery using Aqualase through a 3.2-mm corneal incision. Grade of nucleus, nuclear removal technique, and intraoperative complications were noted. Clinical parameters from postoperative visits were collected., Results: Aqualase is capable of removing cataracts up to nuclear sclerosis 2+ (out of 4) with relative ease. Nuclei graded 2+ or greater were technically more difficult and conversion to ultrasound phacoemulsification was required in one case. Two posterior capsule ruptures occurred: one during nucleus removal (contact with the tip while aspirating without Aqualase) and one unrelated to Aqualase during aspiration of cortex. Of 25 patients seen on the first postoperative day, 22 had a clear cornea. A total of 96% patients without preoperative comorbidity achieved 6/9 or better postoperatively. One patient had transient postoperative uveitis., Conclusions: Removal of softer cataracts with Aqualase has the theoretical advantage over phacoemulsification, by carrying less risk to the posterior capsule, since the handpiece has a smooth polymer tip that has no mechanical motion inside the eye. However, the tip should not be considered entirely capsule-friendly, as rupture is possible with the foot-pedal in position two (aspiration only). Although certain adjustments to the technique are required, the method is similar enough to phacoemulsification to ensure a brief learning curve. With increasingly firm cataracts, Aqualase becomes less effective and ultrasound phacoemulsification is still superior for such cases, in our experience.

HLA-B*51 is known to be associated with Behcet's disease (BD) in many ethnic groups. The pathogenic gene, however, may lie close to the HLA-B locus and therefore be in linkage disequilibrium with HLA-B*51. On the basis of the proximity of MIC genes to HLA-B, their expression pattern and their affinity for the activating NKG2D receptor on natural killer (NK) cells and gammadelta T cells, these molecules have been postulated as susceptibility factors in BD. DNA from 56 western European Caucasians with BD and 90 Caucasian controls were analysed by polymerase chain reaction using allele-specific primers for MICA and MICB alleles. An increased allele frequency of MICA*009 was found in the BD patient group (25.0%) when compared with the controls (7.2%). This was associated with a corresponding decrease in MICA*008 in the BD patients (36.6%) compared with the controls (46.7%), which was not significant. MICA*009 was strongly associated with the presence of HLA-B*51 in patients and controls. No significant difference in frequency of MICB alleles was found between patients and controls. Both HLA-B*51 and MICA*009 are strongly associated with BD in a pure Caucasian BD patient group, and the two alleles are in linkage disequilibrium. No MICB allele was found to associate significantly with the disease, an unexpected finding considering the close proximity of the MICA and MICB loci. Our results suggest that while MICB does not influence the development of BD, polymorphisms in MICA may be pathogenic, perhaps through the interaction with NK and gammadelta T cells.

Aims: To assess visual function, vision related quality of life (VR-QOL), and general health related quality of life (HR-QOL) in intermediate uveitis (IU)., Methods: VR-QOL and HR-QOL were evaluated in 42 patients with IU using the VCM1 and SF-36 questionnaires, respectively. LogMAR visual acuity (VA), Pelli-Robson contrast sensitivity (CS), Farnsworth-Munsell 100 hue colour vision (CV), and Estermann visual field (VF) were recorded monocularly and binocularly., Results: Median (interquartile range) visual acuity (VA) and CS of 72 affected eyes were 0.1 (0.015-0.3) and 1.55 (1.35-1.65), respectively. 9.5% of patients had a VCM1 score of more than 2.0, indicating "more than a little" concern over vision. Worse eye VA (p=0.045) and CS (p=0.042) were predictive of a VCM1 score of more than 2.0 independently of age, sex, uveitis duration, laterality and activity, systemic uveitis therapy, and medical co-morbidity. The physical and mental component summary scores of the SF-36 were significantly worse in those who reported significant impairment of vision on the VCM1 than those who did not., Conclusions: The majority of patients with IU maintain good visual function and quality of life. VR-QOL impairment in IU correlates with vision in the worse eye and is associated with impaired HR-QOL.

Purpose: To determine architectural differences between classic and occult choroidal neovascularization (CNV) in vivo., Design: Prospective observational case series., Methods: Twenty-two patients with acute CNV underwent fluorescein angiography and optical coherence tomography (OCT), which were analyzed by separate blinded observers., Results: In 87.5% of angiographically labeled "classic" CNV a discreet subretinal lesion corresponding to the neovascular complex could be seen above and separate to the retinal pigment epithelium on OCT. This was found in only 13.3% of "occult" CNV., Conclusion: With the latest commercially available OCT equipment it is now possible to confirm in vivo the previously proposed anatomic differences between fluorescein angiographically labeled classic and occult CNV. Classic CNV appear to grow predominantly in the subretinal space, whereas the majority of occult lesions do not. Optical coherence tomography features of CNV may correlate with response to photodynamic therapy or angiostatic treatments, as well as predicting the success of surgical removal.

Background: Given the presence of neural progenitor cells (NPC) in the retina of other species capable of differentiating into multiple neural components, the authors report the presence of NPC in the adult human retina. A resident population of NPC suggests that the retina may constitutively replace neurons, photoreceptors, and glia., Methods: Adult human postmortem retinal explants and cell suspensions were used to generate cells in tissue culture that display the features of NPC. The phenotype of cells and differentiation into neurons was determined by immunocytochemistry. Dividing cells were labelled with 5-bromo-2-deoxyuridine (BrdU) and neurospheres were generated and passaged., Results: Cells labelled with nestin, neurofilament M (NFM), rhodopsin, or glial fibrillary acidic protein (GFAP) grew out from explant cultures. BrdU labelling of these cells occurred only with basic fibroblast growth factor (FGF-2). Dissociated retina and pars plana generated primary neurospheres. From primary neurospheres, NPC were passaged to generate secondary neurospheres, neurons, photoreceptors, and glia. BrdU labelling identified dividing cells from neurospheres that differentiated to express NFM and rhodopsin., Conclusion: The adult human retina contains NPC and may have the potential to replace neurons and photoreceptors. This has implications for the pathogenesis and treatment of retinal disorders and degenerations, including glaucoma, and those disorders associated with retinal scarring.

Transgenic hairy roots of Catharanthus roseus were established with glucocorticoid inducible tryptophan decarboxylase (TDC) expression alone or in combination with inducible expression of a feedback-resistant anthranilate synthase alpha subunit (ASalpha) from Arabidopsis. Northern blot analysis confirmed transcription of the anthranilate synthase gene upon induction in the double line (TDC+ASalpha) and in vitro enzyme assays confirmed increased resistance to feedback inhibition by tryptophan. In TDC enzyme assays, increases of 48% and 87% in the TDC and double lines, respectively, were noted. Although the TDC line showed no significant increase in tryptamine levels on induction, induction of the double line resulted in increases in tryptamine levels of as much as six-fold for a 3 day late exponential induction. Downstream effects on alkaloids were noted in the TDC line where serpentine specific yields increased as much as 129% on induction. No effects on measured alkaloids were noted in the double line, but the two clones have very different basal alkaloid biosynthetic capacities. Within this study, the engineering of the indole pathway in C. roseus hairy roots is reported, and the role of the indole pathway in alkaloid biosynthesis explored.

Different plant species produce a variety of terpenoid indole alkaloids, which are of interest as plant defensive secondary metabolites and as valuable pharmaceuticals. Although significant progress has been made, the mechanisms regulating the levels of this important class of compounds require continued elucidation. Previous precursor feeding studies have indicated that alkaloid accumulation can be improved during the exponential growth phase of hairy root cultures through enhanced tryptophan availability. To test this relationship, transgenic hairy root cultures of Catharanthus roseus were established with a glucocorticoid-inducible promoter controlling the expression of an Arabidopsis feedback-resistant anthranilate synthase alpha subunit. Enzyme assays demonstrated that the Arabidopsis alpha subunit is compatible with the native beta subunit and that anthranilate synthase activity is more resistant to tryptophan inhibition in induced than in uninduced extracts. The metabolic effects of expressing the feedback-resistant anthranilate synthase alpha subunit were also dramatic. Over a 6-day induction period during the late exponential growth phase, tryptophan and tryptamine specific yields increased from almost undetectable levels to 2.5 mg/g dry weight and from 25 microg/g to 267 microg/g dry weight, respectively. The greater than 300-fold increase in tryptophan levels observed in these studies under certain induction conditions compares favorably with the fold increases obtained in previous constitutive expression studies. Despite the large increases in tryptophan and tryptamine, the levels of most terpenoid indole alkaloids were not significantly altered, with the exception of lochnericine, which increased 81% after a 3-day induction period. These results suggest that terpenoid indole alkaloid levels are tightly controlled., (Copyright 2004 Wiley Periodicals, Inc.)

Purpose: Minocycline, a semi-synthetic tetracycline antibiotic is reported to be neuroprotective in degenerative and ischaemic models of central nervous system disease, via mechanisms involving suppression of both cytotoxic microglial activity and caspase-dependent apoptosis. We have investigated the effect of minocycline treatment on a mouse model of retinitis pigmentosa, an inherited photoreceptor neurodegenerative disorder, and contrasted this with the effect of depleting retinal microglia using liposomal clodronate., Methods: rds mice were treated intraperitoneally from the second postnatal day (P2) with either daily minocycline until P16, P18, P21, P24 and P27 or alternative day clodronate liposomes until P16. Immunohistochemical and immunofluorescent methods were applied for the detection of microglia (F4/80) and apoptosis (TUNEL and caspase 3 activation)., Results: Photoreceptor apoptosis was delayed by minocycline treatment but not, ultimately, prevented. Markedly reduced expression of activated caspase 3 was observed in photoreceptors at the early time point, corresponding with the reduced level of apoptosis. Delayed photoreceptor apoptosis due to minocycline treatment was associated with a 50% reduction in the numbers of microglia at early timepoints. Liposomal clodronate treatment also resulted in a marked reduction in the number of microglia (63% reduction in microglia), but in contrast to minocycline treatment, this had no effect on photoreceptor apoptosis., Conclusions: Minocycline appears to delay photoreceptor apoptosis through a microglia-independent action. Although microglial cytotoxicity has been implicated during other models of neurodegeneration, microglia are unlikely to play such a role in this model of photoreceptor dystrophy.

Retinal vasculitis is a rare, but potentially blinding intraocular inflammatory condition with diverse aetiology. Although commonly idiopathic, it has a strong association with systemic inflammatory diseases known to involve other areas of the central nervous system, most notably Behcet's disease, sarcoidosis, systemic lupus erythematosis and multiple sclerosis. This article describes the clinicopathologic features of retinal vasculitis and its visually damaging sequelae, reviewing available human histopathologic studies and work with experimental models to discuss the pathogenesis and immunopathology. Evidence indicates that noninfective retinal vasculitis is an autoimmune condition that may be induced by antecedent infection with microbes cross-reacting with putative autoantigens, influenced by genetic susceptibility of both HLA associations and cytokine polymorphisms. The growing understanding of the cellular mechanisms involved in the effector immune response is already providing a rationale for more specific therapeutic approaches.

Purpose: The retina contains a rich network of myeloid-derived cells (microglia) within the retinal parenchyma and surrounding vessels. Their response and behavior during inflammation and neurodegeneration remain largely undefined. In the present study, the behavior of microglia was closely examined during the onset of photoreceptor degeneration in the rds mouse, to assess their role in photoreceptor apoptosis. The results may have relevance to similar degeneration in humans (retinitis pigmentosa)., Methods: Retinas from rds and wild-type CBA mice aged 8, 14, 16, 17, 19, 21, 30, and 40 days were examined immunohistochemically, with antibodies to macrophage cell surface markers, inducible nitric oxide synthase (iNOS), and proliferating cell nuclear antigen (PCNA), during the most active phase of the disease. TUNEL was used to assess photoreceptor apoptosis., Results: In the rds mouse, microglia proliferated in situ (PCNA), migrated to the subretinal space, and adopted an activated phenotype. Maximum microglial cells occurred at postnatal day (P)21, 5 days after the peak in photoreceptor apoptosis (P16). Microglia did not express iNOS, and nitrotyrosine was absent. Sialoadhesin was expressed on microglia from P14, and expression was greatest at P21., Conclusions: During retinal degeneration, microglia are activated and express sialoadhesin. The temporal relationship between photoreceptor apoptosis and microglial response suggests that microglia are not responsible for the initial wave of photoreceptor death, and this is corroborated by the absence of iNOS and nitrotyrosine. Expression of sialoadhesin may indicate blood-retinal barrier breakdown, which has immune implications for subretinal gene therapeutic strategies.

The 1-deoxy-D-xylulose-5-phosphate (DXP) pathway (non-mevalonate pathway) leading to terpenoids via isopentenyl diphosphate (IPP) has been shown to occur in most bacteria and in all higher plants. Treatment with the antibiotic fosmidomycin, a specific inhibitor of DXP reductoisomerase, considerably inhibited the accumulation of the alkaloids ajmalicine, tabersonine, and lochnericine by Catharanthus roseus hairy root cultures in the exponential growth phase. However, fosmidomycin did not significantly affect alkaloid levels in stationary phase hairy root cultures. Feeding with 1-deoxy-D-xylulose, 10-hydroxygeraniol, or loganin resulted in significant increases in alkaloid production by exponential phase hairy root cultures. These results suggest that the DXP pathway is a major provider of carbon for the monoterpenoid pathway leading to the formation of indole alkaloids in C. roseus hairy roots in the exponential phase.

Transgenic hairy root cultures of Catharanthus roseus were established with a glucocorticoid-inducible promoter controlling the expression of green fluorescent protein (GFP), and GFP expression was characterized. The inducible system shows a tightly controlled, reversible, and dosage-dependent response to the glucocorticoid dexamethasone in C. roseus hairy roots. Full induction was noted after 12-18 h in the mature regions of the root tips and after 6 h in the meristem tissue. Upon removal of the inducing agent, GFP expression declined to undetectable levels in the mature tissues after 24 h and in the meristem after 48 h. Although no dosage-dependent response was noted in the meristem region, such a response was apparent in the mature region of the tip and verified by quantitative GFP analysis. The inducible promoter system allowed quantitative control of GFP expression between 0.01 and 10 microM dexamethasone with saturation occurring at higher levels. Using GFP as a model system allowed demonstration of the ability to control temporal and quantitative gene expression with the glucocorticoid-inducible promoter in transgenic C. roseus hairy roots.

Aim: A case of severe normal tension glaucoma is reported in a trumpet player, along with a study investigating the association between glaucoma and raised intrathoracic pressure, using measurements of pulsatile ocular blood flow (POBF)., Methods: Three patient groups were studied; normals (n = 34), untreated primary open angle glaucoma (POAG) (n = 20), and untreated normal tension glaucoma (NTG) (n = 22), with a total of 76 patients who underwent measurements of POBF using the OBF pneumotonometer at rest and while forcibly exhaling through a mouthpiece connected to a mercury manometer (30 mm Hg) (Valsalva manoeuvre)., Results: POBF fell during Valsalva in all groups with the greatest predictor being the resting value of POBF. There was no evidence of significant differences in the mean change in POBF occurring during the Valsalva manoeuvre for the three groups studied before or after adjusting for the sex, the resting POBF, and the resting IOP of the patients (p = 0.294 and p = 0.542, respectively). However, statistically significant associations were found between the change in POBF and sex (p = 0.049), resting POBF (p<0.0001) and resting IOP (p = 0.032). Males had a greater drop, on average, in POBF during Valsalva manoeuvre than females after adjusting for the other factors. Additionally, there was a significant difference in the mean change in IOP during Valsalva for the three groups (p = 0.002), with the difference occurring between the normal and POAG groups (p<0.005). The POAG group had, on average, a drop in IOP during Valsalva, while the other two groups had an increase in IOP. Also noted was a significant difference in the distributions of the risk factors among the three groups (p = 0.002)., Conclusions: This study demonstrates no difference between groups with respect to resting or Valsalva POBF, but does demonstrate a possible trend with respect to IOP, with a drop in IOP occurring during Valsalva in the POAG group. There is, however, much variability in the data left unexplained by our models. Thus, unfortunately, we cannot advise our trumpet player whether his NTG is directly related to his trumpet playing.

Alkaloids purified from plants provide many pharmacologically active compounds, including leading chemotherapy drugs. As is generally true of secondary metabolites, overall productivity is low, making commercial production expensive. Alternative production methods remain impractical, leaving the plant as the best source for these valuable chemicals. Recently, significant progress in characterizing the biosynthetic pathways leading to various alkaloids has been made, and a number of relevant genes have been cloned. Metabolic engineering employing such genes provides a promising technology for improved productivity in plant cell cultures, plant tissue cultures, or intact plants. In exploring solutions though, metabolic engineers must be careful to recognize the limitations inherent in designing plant systems.

Purpose: To evaluate patient satisfaction with and effectiveness of the 'one-stop' cataract service at the Bristol Eye Hospital., Methods: One hundred and ninety patients referred with a view to cataract surgery were selected to attend 24 'one-stop' clinics between February 1997 and August 1999. Patients had initial assessment in the morning with suitable cases undergoing surgery during the afternoon. Forty 'one-stop' patients and 40 patients undergoing traditional surgery (three separate visits: initial clinic, pre-assessment and surgery) were surveyed by a questionnaire to determine their satisfaction with the service., Results: One hundred and fifty-six of 190 (82.1%) patients invited to one-stop clinics underwent surgery the same day. Questionnaires were returned by 72 patients (35 'one-stop' and 37 control patients). Twenty-eight (80%) 'one stop' patients were 'very satisfied' and 7 (20%) were 'satisfied' with the time allowed to decide upon undergoing surgery, compared with 21 (57%) and 16 (43%) respectively in the control group (p = 0.06, chi-square, not significant). Overall satisfaction with the service was similar between the groups, with 33 (94%) of 'one-stop' and 35 (95%) of control patients considering the service 'very good'., Conclusions: There is a high level of patient satisfaction with a 'one-stop' cataract service which reduces hospital attendance from three visits to one. Potential problems include wasted theatre space when there is a shortfall of suitable cases on the day.

Radioimmunoassay was used to determine alpha-fetoprotein (AFP), albumin, and transferrin production (ng/10(5) cells/24 h) by two cell lines (7777 and 8994) derived from chemically induced rat hepatomas. alpha-Fetoprotein production was high (2000 to 4400) in 7777, but was very low (0.2 to 0.4) in 8994. Albumin production varied from 0.4-0.8 (7777) to 14-26 (8994). Both lines produced substantial amounts of transferrin (180 to 240 by 7777 and 29 to 42 by 8994). Addition of dimethyl sulfoxide (DMSO, 1 to 4%) or sodium butyrate (BA, 0.5 to 2.0 mM) to the medium inhibited growth in both lines, but 8994 was more sensitive to these agents than 7777. Dimethyl sulfoxide treatment (2 to 4%) resulted in a dose-related decrease (less than 10% of control at 4% DMSO) in AFP, albumin, and transferrin production by 7777, but in 8994, DMSO (1 to 2%) resulted in an increase (up to sixfold) in albumin and transferrin production, without affecting AFP production. By contrast, BA (2 to 4 mM) stimulated the production of all three proteins in both lines, most notably that of albumin (up to sixfold) by 7777 and that of AFP (up to 20-fold) by 8994. It is concluded that both DMSO and BA can enhance the expression of differentiated functions of the hepatoma cell, and that DMSO at the same time can suppress the expression of an oncofetal function. However, neither DMSO nor BA is selective in its effects on specific genes (i.e., normal, adult vs. oncofetal genes), and it appears that their effects may be the result of a more general phenomenon, the expression of which may be related to the stage of differentiation of the cell.