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Purpose: We assess the safety and effectiveness of intranasal neurostimulation to promote tear production via the nasolacrimal pathway in subjects with dry eye disease. Methods: A multicenter, randomized, controlled, double-masked pilot study was conducted in adults with dry eye diagnosis and at least one eye with corneal fluorescein staining ≥2 in at least one region or a sum of all regions ≥5 (National Eye Institute grading), basal Schirmer test score ≤10 mm, a cotton-swab stimulated Schirmer score ≥7 mm higher, and an Ocular Surface Disease Index score ≥23. Subjects were randomized to receive active intranasal neurostimulation or sham control intranasal stimulation 4 to 8 times per day. Assessments were scheduled before (unstimulated) and during (stimulated) device application at days 0, 7, 14, 30, and 90. The primary effectiveness endpoint was stimulation-induced change in Schirmer test (with anesthesia) score. Primary safety measure was incidence of device-related adverse events (AEs). Results: Fifty-eight subjects were randomized at nine sites in Australia and New Zealand; 56 completed the 90-day study. Stimulation-induced change in Schirmer score was significantly greater with active intranasal (mean ± SEM, 9.0 ± 2.0) than sham control intranasal stimulation (0.4 ± 0.6; P < 0.001) at day 90. Similar results were observed at days 0, 7, 14, and 30 (P < 0.001). No serious device-related AEs were observed. Mild nosebleed, the most common device-related AE, was reported in five (16.7%) subjects. Conclusions: Intranasal neurostimulation was effective in inducing acute tear production after 90 days of use and generally was well tolerated in subjects with dry eye disease.

Purpose To define and characterize a novel pre-Descemet's layer in the human cornea. Design Clinical and experimental study. Participants We included 31 human donor sclerocorneal discs, including 6 controls (mean age, 77.7 years). Methods Air was injected into the stroma of donor whole globes (n = 4) and sclerocorneal discs (n = 21) as in the clinical deep anterior lamellar keratoplasty procedure with the big bubble (BB) technique. The following experiments were performed: (1) creation of BB followed by peeling of the Descemet's membrane (DM); (2) peeling off of the DM followed by creation of the BB, and (3) creation of the BB and continued inflation until the bubble popped to measure the popping pressure. Tissue obtained from these experiments was subjected to histologic examination. Main Outcome Measures Demonstration of a novel pre-Descemet's layer (Dua's layer) in the human cornea. Results Three types of BB were obtained. Type-1, is a well-circumscribed, central dome–shaped elevation up to 8.5 mm in diameter (n = 14). Type-2, is a thin-walled, large BB of maximum 10.5 mm diameter, which always started at the periphery, enlarging centrally to form a large BB (n = 5), and a mixed type (n = 3). With type-1 BB, unlike type-2 BB, it was possible to peel off DM completely without deflating the BB, indicating the presence of an additional layer of tissue. A type-1 BB could be created after first peeling off the DM (n = 5), confirming that DM was not essential to create a type-1 BB. The popping pressure was 1.45 bar and 0.6 bar for type-1 BB and type-2 BB, respectively. Histology confirmed that the cleavage occurred beyond the last row of keratocytes. This layer was acellular, measured 10.15±3.6 microns composed of 5 to 8 lamellae of predominantly type-1 collagen bundles arranged in transverse, longitudinal, and oblique directions. Conclusions There exists a novel, well-defined, acellular, strong layer in the pre-Descemet's cornea. This separates along the last row of keratocytes in most cases performed with the BB technique. Its recognition will have considerable impact on posterior corneal surgery and the understanding of corneal biomechanics and posterior corneal pathology such as acute hydrops, Descematocele and pre-Descemet's dystrophies. Financial Disclosure(s) The authors have no proprietary or commercial interest in any materials discussed in this article.

Background Research on harm reduction has typically focused on broad-based or organisational strategies such as needle exchange and opiate substitute programmes. Less attention has been paid to the self-directed harm reduction practices of substance users themselves. Few studies have focused on sexual minority populations such as gay and bisexual men and fewer still on the marginalised groups that constitute these populations. This paper identifies self-directed harm reduction strategies among substance using ethno-racially diverse gay and bisexual men. Methods This article presents findings from the Party Drugs Study in Toronto's gay dance club scene, a community-based qualitative study in Toronto, Canada. We present a thematic analysis of interviews with 43 gay and bisexual men from diverse ethno-racial backgrounds about their substance use in the gay dance club scene. Findings We identify five self-directed harm reduction strategies: rationing, controlling or avoiding mixing, controlling quality, maintaining a healthy lifestyle, and following guidelines during substance use. Conclusions We discuss our findings in relation to prior research and to critical theory. We suggest that drug users' awareness of possible harm, and their personal investment in harm reduction, constitute a viable platform from which community-based and public health organisations may promote and strengthen harm reduction among gay and bisexual men from ethno-racially diverse backgrounds.

Purpose: To report on the 12-month follow-up of 77 eyes with moderate to high myopic astigmatism implanted with toric implantable collamer lenses (ICLs). Methods: Retrospective case-note review of 77 eyes from 42 patients undergoing toric ICL placement by one surgeon. Preoperative mean spherical equivalent −2.50 dioptres (D) to −15.00 D myopia and 1.00 D to 7.00 D astigmatism. Results: At 12 months, mean manifest refractive cylinder (MRC) decreased 81% from 2.38 D to 0.44 D. MRC within 1.00 D occurred in 99% (76/77) of eyes, whereas 86% (66/77) had MRC within 0.75 D. 99% (76/77) had postoperative best-corrected visual acuity (BCVA) better than or equal to preoperative values, whereas 78% (60/77) gained up to one line BCVA and 1% (1/77) lost one line BCVA. Uncorrected binocular vision of 6/6 or better occurred in 90% (38/42) of patients compared with binocular BCVA of 6/6 or better in 67% (28/42) preoperatively. One ICL was replaced due to low vaulting. Two eyes with astigmatism of 3.25 D and 3.50 D received subsequent laser in situ keratomileusis (LASIK) to reduce residual small refractive errors. Indications for ICL were: myopia too high for LASIK (73%), cornea too thin for LASIK (44%) and contact lens intolerance (33%). Night halos were reported in 10% (8/77) of eyes at 12 months. One ICL was removed due to unrecognized preoperative glaucoma. There were no cases of cataract formation, or endophthalmitis. Conclusion: This study is the largest reported series of toric ICL implantation in New Zealand. It supports the safety, efficacy and predictability of toric ICLs to treat myopic astigmatism.

Background: To report on 1-year follow up of a series of 126 eyes implanted with a spherical implantable collamer lens (ICL). Methods: Retrospective case note review of 126 eyes (68 patients) undergoing ICL placement by a single surgeon (TG) to correct preoperative mean spherical equivalents between +8.88 and −20.50 D. Main outcome measures included uncorrected visual acuity (UCVA), refraction, best spectacle-corrected VA, unaided binocular vision, adverse events, operative/postoperative complications and patient symptoms. Results: Complete data were available at 1 year on 121 of the 126 eyes (65 out of 68 patients). At 12 months 86% (104/121) had postoperative UCVA better than or equal to preoperative best spectacle-corrected VA, and 98% (118/121) had UCVA within one line of preoperative best spectacle-corrected VA. Ninety-six per cent (116/121) had postoperative UCVA of 6/9 or better, and 100% had postoperative best-corrected visual acuity of 6/12 or better. Ninety-eight per cent (64/65) patients had unaided binocular vision at 1 year of 6/7.5 or better. Average myopia treated was −8.83 D (116 patients), and average hyperopia +7.14 D (10 patients). Two ICLs were replaced because of high vaulting; one eye had increased intraocular pressure that resolved with smaller ICL placement. Four patients had visually insignificant pigment deposits on the ICL at 3/12, but no pigmentary glaucoma. Five patients noticed haloes around lights at night, none preventing driving. Conclusions: One-year results from this study, the largest reported clinical investigation of the use of ICLs in New Zealand, support the safety, efficacy and predictability of ICL to treat both hyperopic and myopic spherical refractive errors.

This paper is intended as an historical document describing the entry and success of the Decca Radar Company into the production of Doppler-based navigation units. It is based on an original personal paper written in about 1970 by Trevor Gray, then Head of Doppler Department, Decca Radar, updated and edited with additions by Walter Blanchard (past President of the RIN) assisted by Trevor Gray in May 2007. The paper first covers the basic theory and principles of Doppler radar systems, continues through the setting up of the Decca Doppler Department in the mid-1950s and the production of the first transistor-based Doppler system. Success led to the development of both civil and military Doppler systems culminating in the Decca Doppler 70 Series which was fitted to some 30 types of aircraft and helicopters, worldwide.

Over a period of approximately 7 months, multiple subepithelial spots were noted in one or both corneas of some patients examined 2 to 6 months after laser in situ keratomileusis (LASIK). The lesions appeared identical to adenovirus keratitis. The eyes were quiet, had good vision, and no patient had a history of adenoviral keratoconjunctivitis. In all eyes, the lesions resolved spontaneously or with topical steroid eyedrops. All investigations were negative for adenovirus. Corneal laser scanning in vivo confocal microscopy revealed Langerhans cells in the epithelium, which disappeared after the lesions resolved.

The process of corneal epithelial regeneration arising from progenitor or stem cells in the limbal basal region is a well‐accepted physiological phenomenon. This has been based on the large amount of circumstantial laboratory and clinical evidence.1,2 No reliable and specific marker for limbal stem cells exists and work continues in this area. More recently it has been recognised that adult stem cell reside in specific areas or niches.3 These provide the necessary micro‐environment that preserves the stemness of stem cells through a variety of mechanisms.4 Niches have been well characterised in the bulge region of the hair follicle and in the bone marrow for haematopoetic stem cells. However, even though we accept that putative corneal epithelial stem cells reside in the limbal basal cells (and this has been characterised extensively5), no discrete or specific region within the basal limbus has been identified as a niche per se. Recently our group made the novel discovery that in some parts of the limbus, solid cords of epithelial cells extend from the peripheral end of the limbal palisades into the underlying stroma. These were termed the limbal epithelial crypt (LEC).6 Our preliminary work indicated that LECs were few in number, extended into the limbal stroma anteriorly, posteriorly or circumferentially and housed cells that were ABCG‐2 and CK14 positive. We therefore hypothesised that the LEC may act as a putative stem cell niche. The main purpose of this study was to further characterise the anatomy of the LEC in the human eye with the intention that this would yield clues to its role as a putative stem cell niche. In doing so, our objectives were to determine the frequency and distribution of LEC, characterise the immunophenotype of the cells within the LEC and study their ultra structural features with particular reference to the size, morphology, intercellular connections and basal attachments.

BACKGROUND DISC-HSV is a replication incompetent herpes simplex virus that is a highly efficient vector for the transduction of genes in vivo and in vitro. We examine the ability of DISC-HSV to infect human prostate cancer cell-lines and xenograft tumor models, and induce expression of reporter and therapeutic cytokine genes. METHODS Infection was confirmed by cellular staining for the beta-galactosidase reporter gene product, and by EM. Human GM-CSF production following DISC-hGMCSF infection was measured using ELISA. The metabolic activity of infected cells was determined by NADP/NADPH assay. Cell death was estimated by cell-cycle analysis using flow cytometry with propidium iodide staining. RESULTS Infection of DU145, PC3 and LNCaP cells with DISC-HSV was dose dependent. Cells infected with DISC-hGM-CSF released significant levels of hGM-CSF for 3 days. NADP/NADPH assay suggested that infected cells continued to be metabolically active for 3 days post-infection, which was consistent with flow cytometry findings that cell death did not occur within 7 days of infection. Tumor xenografts injected with DISC-HSV expressed beta-galactosidase, and intracellular viral particles were demonstrated using EM. CONCLUSIONS We have previously reported the rejection of established tumors following intra-tumoral injection of DISC-GMCSF. This study demonstrates the ability of DISC-HSV to infect prostate cancer and express GMCSF at significant levels. We suggest that prostate cancer is a potential target for therapy using DISC-HSV containing GM-CSF. Prostate 56: 65–73, 2003. © 2003 Wiley-Liss, Inc.

Objective: This study describes the clinical features, management, and outcome of 19 patients who had severe Acanthamoeba sclerokeratitis (ASK) unresponsive to conventional management, requiring systemic immunosuppression to control disease. Design: Retrospective, noncomparative, interventional case series. Participants: Records of all patients with Acanthamoeba keratitis treated at Moorfields Eye Hospital between 1989 and 2000 were reviewed. From more than 200 patients, 19 who developed ASK treated with systemic immunosuppression were identified. Main Outcome Measures: Visual acuity, level of pain, and degree of inflammation were recorded after immunosuppressive treatment. Results: ASK requiring immunosuppression occurred in 20 eyes of 19 patients (11 males and 8 females). The mean age (mean ± standard deviation) at onset was 38.6 ± 13.2 years. On presentation, best-corrected visual acuity was counting fingers or worse in 11 eyes (55%), 6/18 to 6/60 in 5 eyes (25%), and 6/12 or better in 4 eyes (20%). The mean time between onset of initial symptoms of Acanthamoeba keratitis and commencement of systemic immunosuppression was 4.8 ± 3.5 months. The mean duration of immunosuppression required to control inflammation was 7.2 ± 3.9 months. Severe scleritic pain remained uncontrolled in two patients and resulted in enucleation. Best-corrected visual acuity at final follow-up was counting fingers or worse in eight eyes (40%), 6/18 to 6/60 in six eyes (30%), and 20/40 or better in six eyes (30%). The mean follow-up period after resolution of inflammation was 24.3 ± 20.9 months (range, 0.2-59.7 months). Conclusions: ASK is an uncommon complication of Acanthamoeba keratitis. The scleritis associated with this infection seems to be an immune-mediated response. After topical amebicidal treatment, systemic immunosuppression may be required to control the pain and tissue destruction associated with ASK.

Background The trabecular meshwork (TM) located at the angle of the anterior chamber of the eye contributes to aqueous drainage. A novel layer in the posterior part of the human cornea has recently been reported (the pre-Descemet’s layer (Dua’s layer (PDL)). We examined the peripheral part of this layer in relation to the origin of the TM. Methods The PDL and TM of 19 human donor eyes and one exenterated sample were studied. Samples were examined by light and electron microscopy (EM) for tissue architecture and by immunohistology for four matricellular proteins, five collagen types and CD34. Results EM revealed that beams of collagen emerged from the periphery of PDL on the anterior surface of the Descemet’s membrane and divided and subdivided to continue as the beams of the TM. Long-spacing collagen was seen in the PDL and TM. Trabecular cells (CD34-ve) associated with basement membrane were seen in the peripheral part of the PDL and corresponded to the start of the separation of the collagen lamellae of PDL. Collagen VI was present continuously in PDL and extended into the TM. Matricellular proteins were seen predominantly in the TM with only laminin extending into the periphery of PDL. Conclusions This study provides an insight into the origins of the collagen core of the TM as an extension of the PDL of the cornea. This finding adds to the knowledge base of the TM and cornea and has the potential to impact future research into the TM and glaucoma.

To determine whether preserved human amniotic membrane can reduce corneal haze and keratocyte apoptosis induced by excimer laser photoablation in rabbit corneas.Excimer photoablation was performed bilaterally in 30 rabbits with a 6.0 mm ablation zone and 120 microm depth using the VISX Star laser with the phototherapeutic keratectomy (PTK) mode. One eye was randomly covered by preserved human amniotic membrane secured with 4 interrupted 10-0 nylon sutures, and the other eye served as the control. The amniotic membranes were removed at 1 week, and corneal haze was graded with slitlamp biomicroscopy by 3 masked corneal specialists biweekly for the ensuing 12 weeks until the rabbits were killed. Another 18 rabbits were divided into 4 subgroups and received PTK alone, PTK with membrane, PTK with sham sutures, or PTK with tarsorrhaphy. All eyes were studied histologically, and 3 eyes in each group were studied by in situ terminal deoxynucleotidyl transferase deoxy-UTP-nick end labeling assay at 1, 3, and 7 days and 12 weeks, respectively.A consistent grading of differences in corneal haze scoring between the control corneas and the amniotic-membrane-covered corneas was noted among the 3 masked observers. Organized reticular post-PTK corneal haze peaked at 7 weeks in both groups, and the corneal haze score in the amniotic-membrane-covered group was significantly less than in the control group from 7 to 12 weeks (all P.001). Compared to the control corneas, the amniotic-membrane-covered corneas had less inflammatory response at 1 and 3 days, less keratocyte apoptosis in the ablated anterior corneal stroma at 1, 3, and 7 days (P.001), and less stromal fibroblast cellularity and epithelial hyperplasia at 12 weeks.Amniotic membrane matrix introduced at an early stage of the corneal wound healing process effectively reduced corneal haze induced by excimer laser photoablation in rabbits. Studies linking suppression of apoptosis in the acute wound-healing process with reduction of subsequent corneal scarring may have useful clinical applications.

Background & Aims: In active inflammatory bowel disease (IBD), the intestinal mucosa is infiltrated by polymorphonuclear cells (PMNs), lymphocytes, and monocytes from the systemic circulation. Using an ex vivo model, we have investigated luminally directed migration of cells out of the lamina propria. Methods: Fresh untreated and deepithelialized mucosal samples were studied by electron microscopy. Cells migrating out of the lamina propria were investigated by immunohistochemistry and fluorescence-activated cell sorter analysis. Results: In intact IBD mucosal samples, tunnels containing cells were prominent in the lamina propria matrix, and PMNs, but not other cell types, were prominent in the epithelium. In deepithelialized mucosal samples, the basement membrane was either destroyed or contained numerous large pores. During culture of deepithelialized mucosal samples, many cells (3.3 [±0.8] × 10 5 · g tissue −1 · h −1 ) migrated out of the lamina propria via basement membrane pores. PMNs and eosinophils were prominent during the first 3 hours of culture, but T cells predominated thereafter. Macrophages also migrated, but B cells were the minority population ( Conclusions: In active IBD mucosa with an intact epithelium, luminally directed migration of lamina propria cells is restricted mainly to PMNs. After loss of the epithelium, other cell types also migrate into the lumen via numerous, large, basement membrane pores. GASTROENTEROLOGY 1998;115:841-848

Epithelial cells are the first point of host contact for invasive intestinal pathogens and may initiate mucosal inflammatory responses via production of proinflammatory cytokines and mediators. The aim of the present study was to investigate in vitro the initial invasion of a parasitic nematode ( Trichinella spiralis ), to measure the early production of specific epithelial cytokines and inflammatory mediators after invasion, and to compare these responses with those to invasive bacteria. Monolayers of human colonic epithelial cell lines (HT29, T84, and Caco-2) were infected by T. spiralis or Listeria monocytogenes . Bile-activated infective larvae of T. spiralis invaded and migrated into the epithelial cell monolayers, leaving trails of dead cells. Transmission electron microscopy studies of damaged cells along the trail showed a progressive increase in size, disruption of cell membranes, loss or dilution of cytoplasmic proteins, and swelling of mitochondria and nuclei. However, no nuclear fragmentation was observed. With reverse transcription-PCR and an enzyme-linked oligonucleotide chemiluminescent assay, mRNA transcripts of interleukin-1β (IL-1β), IL-8, and epithelial neutrophil-activating peptide 78 were shown to increase in epithelial cells invaded by T. spiralis or L. monocytogenes , but only L. monocytogenes elicited increased inducible nitric oxide synthase (iNOS) mRNA. No increase in tumor necrosis factor alpha or transforming growth factor β mRNA was seen after T. spiralis invasion. Increased levels of IL-8 were also released from the basolateral surfaces of infected monolayers as detected by sandwich enzyme-linked immunosorbent assay. Induction and secretion of proinflammatory cytokines in epithelial cells after nematode or bacterial invasion may initiate the acute inflammatory response of the small intestine. The upregulation of iNOS in bacterial infections may contribute to mucosal defense and may also be associated with subsequent cell death, whereas different mechanisms appear to operate after nematode invasion.

Objectives: To establish the point prevalence of pathological laughing and crying (PLC) in multiple sclerosis (MS). To define associated neurological, emotional, and cognitive correlates of PLC. Design: A consecutive sample of 152 patients with clinically or laboratory definite MS were screened for PLC, defined as sudden, involuntary displays of laughing or crying or both, without associated subjective feelings of depression or euphoria. Thereafter, a case-control design was followed with patients with PLC matched to patients with MS without PLC on age, gender, physical disability (Expanded Disability Status Scale), duration of MS, and premorbid IQ. Setting: An MS outpatient clinic, the population representative of a large urban catchment area. Patients: Fifteen of 152 patients had PLC, 11 of whom (mean [SD] age, 43.7 [8.3] years, 7 women) agreed to further testing. Thirteen patients with MS without PLC acted as controls. Main Outcome Measures: Neurological examination, Pathological Laughter and Crying Scale, Hospital Anxiety and Depression Scale, 28-item General Health Questionnaire, and the Wechsler Adult Intelligence Scale-Revised. Results: The point prevalence of PLC in MS was 10%. Patients had a mean Expanded Disability Status Scale score of 6.5, had had MS for a mean (SD) of 10 (5.8) years, and had entered a chronic-progressive phase of their illness. Pathological laughing and crying was not associated with disease exacerbations. Compared with controls, patients were not more depressed or anxious, but had a greater decline in IQ. Conclusions: Pathological laughing and crying as distinct from emotional lability affects 1 in 10 patients with MS. It occurs in severely physically disabled patients, generally with long-standing disease. The presence of cognitive deficits relative to controls implies more extensive brain involvement.

Clostridium difficile is the aetiological agent of pseudomembranous colitis, and animal studies suggest the essential role of secreted toxin A in inducing disease. This study examined the biological responses to toxin A by human intestinal epithelial cells. Confluent monolayers of Caco2, HT29, and T84 cells and primary epithelial cells in organ cultures of human colonic biopsy specimens and after detachment with EDTA were studied. Interleukin 8 was assayed using enzyme linked immunosorbent assay (ELISA). Purified C difficile toxin A induced cell rounding and detachment of monolayers of the epithelial cell lines. Cells in detached monolayers initially remained viable while adherent to each other. Subsequently, an increasing number of apoptotic cells appeared in suspension. Exposure to toxin A for 24 hours induced interleukin 8 production in T84 and HT29 cells. Toxin A also induced epithelial cell rounding, detachment, and apoptosis in organ cultures of human colonic biopsy specimens. During culture (in medium only), EDTA detached colonic epithelial cells produced interleukin 8 and cell death occurred by apoptosis. Colonic disease by C difficile may be initiated by toxin A mediated induction of epithelial cell interleukin 8 production and apoptosis after cell detachment from the basement membrane. Studies on isolated (toxin untreated) colonic epithelial cells suggest that interleukin 8 production and apoptosis occur as a consequence of cell injury and detachment.

Purpose To report in vivo confocal microscopic (IVCM) features in corneal edema supported by histopathologic correlation. Methods This was an observational study with evaluation of diagnostic technology. Twenty patients with clinically diagnosed corneal edema were involved, including 11 with Fuchs' endothelial dystrophy (FED). All cases, in addition to a control group of six normal eyes, were examined with IVCM before keratoplasty. Four eyes were examined after surgery. Thirteen corneal samples obtained by penetrating keratoplasty were examined by light and/or electron microscopy. IVCM and histopathologic sections were then analyzed for correlation and proper interpretation. Seven patients underwent Descemet's stripping endothelial keratoplasty (DSEK). Results Subepithelial fibroblasts were seen histologically and with IVCM in 7 (53.8%) of 13 full-thickness corneal samples. IVCM alone detected these changes in 11 (55%) subjects before surgery, as well as after postoperative clinical improvement. Other IVCM features included absent (30%) or reduced (70%) subbasal corneal nerves, expanded hyperreflective keratocyte cell bodies, and processes with small vacuoles and large extracellular lacunae (95%), seen on IVCM only. Endothelial changes with polymegathism and reduced cell density were seen in non-FED cases. Conclusions This is the first study in which IVCM features of corneal edema have been compared in detail with histopathologic findings. Subepithelial fibroblasts, reduced subbasal corneal nerves, and stromal keratocyte morphology were well documented in this study. With increasing popularity of DSEK this work supports the role of IVCM in quantitative evaluation of corneal edema in early preoperative stages, as well as after surgery, when the cornea appear clinically, but not histologically, normal.

To report on the 12-month follow-up of 77 eyes with moderate to high myopic astigmatism implanted with toric implantable collamer lenses (ICLs).Retrospective case-note review of 77 eyes from 42 patients undergoing toric ICL placement by one surgeon. Preoperative mean spherical equivalent -2.50 dioptres (D) to -15.00 D myopia and 1.00 D to 7.00 D astigmatism.At 12 months, mean manifest refractive cylinder (MRC) decreased 81% from 2.38 D to 0.44 D. MRC within 1.00 D occurred in 99% (76/77) of eyes, whereas 86% (66/77) had MRC within 0.75 D. 99% (76/77) had postoperative best-corrected visual acuity (BCVA) better than or equal to preoperative values, whereas 78% (60/77) gained up to one line BCVA and 1% (1/77) lost one line BCVA. Uncorrected binocular vision of 6/6 or better occurred in 90% (38/42) of patients compared with binocular BCVA of 6/6 or better in 67% (28/42) preoperatively. One ICL was replaced due to low vaulting. Two eyes with astigmatism of 3.25 D and 3.50 D received subsequent laser in situ keratomileusis (LASIK) to reduce residual small refractive errors. Indications for ICL were: myopia too high for LASIK (73%), cornea too thin for LASIK (44%) and contact lens intolerance (33%). Night halos were reported in 10% (8/77) of eyes at 12 months. One ICL was removed due to unrecognized preoperative glaucoma. There were no cases of cataract formation, or endophthalmitis.This study is the largest reported series of toric ICL implantation in New Zealand. It supports the safety, efficacy and predictability of toric ICLs to treat myopic astigmatism.

Amniotic membrane (AM) has gained increasing popularity as a useful carrier for ex vivo-expanded cells for tissue engineering, particularly in ocular surface reconstruction. However, current methods employed for denuding AM are highly variable, and the consequent effects on the structural and molecular composition of the AM basement membrane (BM) are ambiguous. We compare the effects of the main denuding procedures, and propose a highly effective standardized alternative.AMs preserved for transplantation were denuded using published ethylenediaminetetraacetic acid (EDTA)- and dispase-based methodologies and our novel thermolysin-based procedure. Scanning and transmission electron microscopy and immunohistochemistry, for BM components (collagens IV and VII, laminin 5, and integrins alpha6 and beta4), were used to assess effectiveness of denuding epithelium, whilst maintaining the integrity of the BM.EDTA- and dispase-based denuding techniques resulted in the disaggregation and even destruction of the BM structure and molecular composition. Employing thermolysin effectively denuded epithelium whilst maintaining BM structural and molecular integrity.Current procedures for preparing AM are variable and often ineffective, resulting in nonstandard membranes. Our novel thermolysin-based technique effectively denudes the AM whilst preserving an essentially intact and consistent BM. Therefore, we propose that this novel thermolysin procedure may potentially improve overall generation of tissue-engineered constructs using AM.

Purpose Amniotic Membrane (AM) has gained increasing popularity as a useful carrier for ex-vivo expanded cells for tissue engineering, particularly in ocular surface reconstruction. However, current methods employed for denuding AM are highly variable and the consequent effects on the structural and molecular composition of the AM basement membrane (BM) are ambiguous. We compare the effects of the main denuding procedures, and propose a highly effective standardised alternative. Methods AMs preserved for transplantation were denuded using published EDTA and Dispase-based methodologies and our novel thermolysin-based procedure. Scanning and Transmission electron microscopy and immunohistochemistry, for BM components (Collgens IV, and VII, laminin 5, and inegrins α6 and -β4) were used to assess effectiveness of denuding epithelium, whilst maintaining the integrity of the BM. Results EDTA and Dispase-based denuding techniques resulted in the disaggregation and even destruction of the BM structure and molecular composition. Employing thermolysin effectively denuded epithelium whilst maintaining BM structural and molecular integrity. Conclusion Current procedures for preparing AM are variable and often ineffective, resulting in non standard membranes. Our novel thermolysin-based technique effectively denudes the AM whilst preserving an essentially intact and consistent BM. Therefore, we propose this novel thermolysin procedure may potentially improve overall generation of tissue engineered constructs using AM.

Purpose When transplanted as a graft AM is incorporated into the cornea. Our aim was to ascertain by immunohistology the fate of the incorporated membrane over time. Methods Corneal buttons from 8 eyes treated by AMT for bullous keratopahty and subsequently had penetrating keratoplasty were examined by electron microscopy and by immunohistology with markers for keratocytes (CD34), fibroblasts (vimentin) and myofibroblasts (SMactin). Time from AMT to PK was between 2 to 32 months Results Amnion tissue was covered with stratified corneal epithelium with well-defined hemidesmosomes. At places a fluid cleft was seen between amnion and underlying Bowman’s zone. Transformed keratocytes/fibroblasts could be seen migrating from the anterior stroma, through breaks in the Bowman’s zone, into connective tissue of the amniotic membrane. Immunohistology showed that the cells populating amnion stroma were CD34 negative but positive for vimentin and smooth muscle actin. In 2 samples where corneal transplants were performed more than 1 year following AMT, some cells in the amniotic stroma showed CD34+ staining Conclusion The amniotic basement membrane facilitates epithelial cell migration and adhesion. Corneal stromal keratocytes can migrate through breaks in Bownans zone, into the amniotic tissue thus integrating it with the host. Despite the presence of large fluid clefts between amniotic membrane and Bowman’s zone, the overlying epithelium remained compact. Repopulation of the amniotic stroma by corneal keratocytes allows for rebuilding of corneal stroma with an indication that over time they may revert to the resting keratocyte immunophenotype.

Purpose To evaluate the histologic changes occurring in the transplanted amniotic membrane in human eyes. Design Observational consecutive case series. Participants Seven consecutive patients who underwent amniotic membrane transplantation (AMT) for bullous keratopathy and subsequently had a penetrating keratoplasty (PK). Methods Corneal buttons obtained at PK were examined by light and electron microscopy and by immunohistology with antibodies against CD34 (keratocytes), α smooth muscle actin and vimentin (myofibroblasts and fibroblasts respectively). Time from AMT to PK ranged from 2 to 32 months. Main Outcome Measures Immunophenotypic characteristics of cells populating transplanted amniotic stroma. Results Amniotic tissue was covered with stratified corneal epithelium with well-defined desmosomes and hemidesmosomes. Transformed corneal stroma-derived cells (CSDCs) could be seen migrating from the anterior stroma, through breaks in the Bowman's zone, into connective tissue of the amniotic membrane. Immunohistology showed that the cells populating amniotic stroma were CD34 negative but positive for vimentin and α smooth muscle actin. In 2 samples in which corneal transplants were performed approximately 1 year or more after AMT, some cells in the amniotic stroma showed CD34+ staining. Features of increased metabolic activity and formation of new collagen were seen on electron microscopy. In 2 cases, epithelial cell nests were seen in the amniotic stroma. Conclusions The amniotic basement membrane facilitates epithelial cell migration and adhesion. The amniotic stroma supports CSDCs and epithelial cells. Repopulation of the amniotic stroma by CSDCs migrating through breaks in Bowman's zone integrates the amnion with corneal tissue and allows for rebuilding of corneal stroma. Over time, some CSDCs may revert to the resting keratocyte immunophenotype. Financial Disclosure(s) The author(s) have no proprietary or commercial interest in any materials discussed in this article.

Ubiquitin-positive intraneuronal inclusions are a consistent feature of the major human neurodegenerative diseases, suggesting that dysfunction of the ubiquitin proteasome system is central to disease etiology. Research using inhibitors of the 20S proteasome to model Parkinson's disease is controversial. We report for the first time that specifically 26S proteasomal dysfunction is sufficient to trigger neurodegenerative disease. Here, we describe novel conditional genetic mouse models using the Cre/loxP system to spatially restrict inactivation ofPsmc1(Rpt2/S4) to neurons of either the substantia nigra or forebrain (e.g., cortex, hippocampus, and striatum). PSMC1 is an essential subunit of the 26S proteasome andPsmc1conditional knock-out mice display 26S proteasome depletion in targeted neurons, in which the 20S proteasome is not affected. Impairment of specifically ubiquitin-mediated protein degradation caused intraneuronal Lewy-like inclusions and extensive neurodegeneration in the nigrostriatal pathway and forebrain regions. Ubiquitin and α-synuclein neuropathology was evident, similar to human Lewy bodies, but interestingly, inclusion bodies contained mitochondria. We support this observation by demonstrating mitochondria in an early form of Lewy body (pale body) from Parkinson's disease patients. The results directly confirm that 26S dysfunction in neurons is involved in the pathology of neurodegenerative disease. The model demonstrates that 26S proteasomes are necessary for normal neuronal homeostasis and that 20S proteasome activity is insufficient for neuronal survival. Finally, we are providing the first reproducible genetic platform for identifying new therapeutic targets to slow or prevent neurodegeneration.

Aim: The conjunctiva has a resident population of intraepithelial and stromal immune cells. These cells play an active part in ocular surface defence and disease. Our aim was to study the migration of immune cells in the human conjunctiva, across the basement membrane and to characterize their phenotypes. Methods: Organ cultures of human conjunctival samples, denuded of the epithelium, were maintained for varying time periods. Cells migrating on to the surface were harvested and analysed by flow cytometry. Conjunctival samples were also studied by immunohistology and electron microscopy. Results: A preferential unidirectional migration of immune cells from the stroma, through pores in the basement membrane, towards the surface was noted. Cells migrated through intrastromal channels, communicating with the surface basement membrane pores. CD3+ T cells (76.18%) were the predominant migrating phenotype. The ratio of CD4:CD8 T cells was approximately 4:1 as compared with the control conjunctiva where the ratio was approximately 2:1. Various other phenotypes including NK, NKT and B cells were also detected. Only 8.41% of the migrating population expressed the Human mucosal lymphocyte-1marker of intraepithelial lymphocytes. Conclusions: Immune cells migration at the ocular surface is an active process involving the formation of intrastromal channels, and cell egression through intact basement membrane pores. The preferential migration of CD4 T cells indicates that this is a specific response of conjunctival lymphoid tissue and not a passive movement of cells. A wide range of immune cell phenotypes exist at the ocular surface. This model can serve to test in vitro the effects of injurious agents on the ocular surface.

To examine the evidence that a series of cases of diffuse lamellar keratitis (DLK) after laser in situ keratomileusis (LASIK) was caused by a type of marker pen.Eye Institute, Auckland, New Zealand.During a 10-week period, 522 consecutive LASIK procedures were performed using a 60 Hz IntraLase femtosecond laser (IntraLase Corp.) to create the LASIK flap and a 217Z 100 Hz excimer laser (BauschLomb) to perform the refractive ablation. As standard practice, a marking pen was used to enable accurate flap realignment. Three weeks after a sudden increase in the incidence of DLK was identified, one of the 5 surgeons performed 5 consecutive bilateral cases using the marking pen in the right eyes but not in the left eyes.Of the 522 LASIK cases (119 without marking pen, 403 with marking pen), DLK developed in 49 (9.4%). No eye treated without the marking pen developed DLK; of those in which the marking pen was used, 49 (12.2%) developed DLK (P0.0001, Fischer exact test; odds ratio, 27). In the 5 consecutive bilateral cases in which the marking pen was used in the right eye but not the left eye, 4 right eyes and no left eye developed DLK (P=0.03). Forty-five of the 49 eyes with DLK quickly recovered. The other 4 developed central toxic keratopathy.There is strong statistical evidence that the marking pen was a factor in the occurrence of DLK.

The GORE-TEX (W. L. Gore & Associates, Flagstaff, Arizona) graft was used in humans for about 10 years starting in 1982. During that time thousands of these ligaments were inserted. In our hospital, 162 GORE-TEX cruciate reconstructions were performed between 1986 and 1993 using a standard over-the-top technique. Two of these patients developed inguinal lymphadenopathy secondary to GORE-TEX particulate materials from the ipsilateral knee joint. To our knowledge, this is the first paper describing the above-mentioned complication, which was established by histologic examination, including electron microscopy.

Interferon beta-Ia (Avonex) 30 microg, intramuscular (i.m.), once weekly is efficacious in delaying clinically definite multiple sclerosis (CDMS) following a single demyelinating event (SDE). This study determined the cost effectiveness of Avonex compared to current treatment in delaying the onset of CDMS.A cost-effectiveness analysis (CEA) and cost-utility analysis (CUA) were performed from Ministry of Health (MoH) and societal perspectives. For CEA, the outcome of interest was time spent in the pre-CDMS state, termed monosymptomatic life years (MLY) gained. For CUA, the outcome was quality-adjusted monosymptomatic life years (QAMLY) gained. A Markov model was developed with transitional probabilities and utilities derived from the literature. Costs were reported in 2002 Canadian dollars. Costs and outcomes were discounted at 5%. The time horizon was 12 years for the CEA, and 15 years for the CUA. All uncertainties were tested via univariate and multivariate sensitivity analyses.In the CEA, the incremental cost of Avonex per ILYgained was $53110 and $44789 from MoH and societal perspectives, respectively. In the CUA, the incremental cost of Avonex per QAMLY gained was $227586 and $189286 from MoH and societal perspectives, respectively. Both models were sensitive to the probability of progressing to CDMS and the analytical time horizon. The CUA was sensitive to the utilities value.Avonex may be considered as a reasonably cost-effective approach to treatment of patients experiencing an SDE In addition, the overall incremental cost-effectiveness profile of Avonex improves if treatment is initiated in pre-CDMS rather than waiting until CDMS.

Background. It is well known that portacaval shunting ultimately leads to a decrease in liver volume and hepatic function, but the mechanism is uncertain. The aim of the present study was to evaluate the effect of portacaval shunting (PCS) upon the morphological changes that occur in the liver in rats after port caval. anastomosis. Materials and methods. Sixty-six male rats underwent either PCS (n = 35) or sham operations (n = 31). Hormone levels were determined in blood samples taken just before removal and weighing of the livers. Hematoxylin and eosin-stained sections were used for quantitative morphometric analysis. Apoptosis, mitosis, and cellular organelles also were assessed quantitatively. Results. There was a significant reduction in the liver mass together with testosterone levels in PCS rats in comparison with sham rats. The distance between presinusoidal and postsinusoidal vessels was reduced from 500 mu m in the sham rats to 299 mu m in the PCS rats (P = 0.000001). Within the same group, there was a significant reduction in the area of hepatocyte nuclei in zone 3 in comparison with zone 1. Electron microscopy revealed a highly significant (P = 0.0007) reduction in the membrane-bound cytoplasmic organelles of zone 3 hepatocytes in PCS rats in comparison with the sham rats. Apoptosis was increased in zone 3 in PCS rats (P = 0.00001), whereas in zone 1 of the same group, there was an associated increased in mitosis (P = 0.000001). Overall, the degree of apoptosis was in excess of mitosis, resulting in a general loss of liver mass. Conclusion. Morphometric analysis at cellular and subcellular levels confirms the morphological findings of liver atrophy in PCS rats. The mechanism of atrophy is a complex one. Portacaval shunting leads to hepatic atrophy that, in turn, results in microcirculatory and hormonal changes that further contribute to liver cell loss in this animal model. (C) 2006 Elsevier Inc. All rights reserved.

PURPOSE To establish a relationship between the forward protrusion of the cornea (keratectasia), following successful LASIK, and anterior chamber depth. METHODS One hundred and fifteen eyes of 59 patients who had undergone Zyoptix LASIK and 39 eyes of 21 patients who had undergone PlanoScan LASIK at the Eye Institute, Centre for Refractive Surgery, Remuera, Auckland, New Zealand, were included in the study. The results of Orbscan II acquisitions, taken before and 8.3 weeks +/- 4.0 (mean +/- standard deviation) following the procedure, were analysed to determine the amount of forward protrusion of the central 2 mm diameter of the posterior surface. The apparent keratectasia was then related to the Orbscan II anterior chamber depth (ACD) measurement in a linear mixed model analysis that included 'right or left eye', 'sex', 'type of LASIK procedure', 'change in pachymetry', 'postoperative pachymetry', 'change in anterior corneal curvature', 'postoperative anterior corneal curvature', 'planned ablation depth' and 'time until follow-up' as factors and covariates. RESULTS The mean +/- SD amount of keratectasia determined by Orbscan II was 22.8 +/- 26.8 microm for the Zyoptix group and 16.9 +/- 24.6 microm for the PlanoScan group. The reduction in depth of the anterior chamber was 14.7 +/- 79.3 microm for the Zyoptix group and 18.2 +/- 68.8 microm for the PlanoScan group. The mixed model analysis demonstrated strong relationships with postoperative pachymetry and change in anterior corneal curvature (P < 0.001 and P < 0.001, respectively). Furthermore, the statistical model showed a marked significant difference in the Orbscan II assessment of keratectasia and change in ACD (P < 0.001). CONCLUSION Using the Orbscan II device, the forward protrusion of the posterior corneal surface appears to coincide with a paradoxical reduction in depth of the anterior chamber. Although the keratectasia found in this study is accompanied by thin postoperative corneas and larger central pachymetry change, the contradictory decrease in anterior chamber depth creates uncertainty of measurement, and of ectasia as a mechanism of regression.

Purpose To investigate the efficacy of alcohol delamination in the management of recurrent corneal erosions (RCEs). Design Prospective single-center consecutive descriptive case series. Participants Twelve patients with RCEs who did not respond to conservative management were treated with alcohol delamination. Methods A consecutive case series of 12 patients with RCEs who did not respond to conservative management were treated by alcohol delamination. A pain score was generated based on a visual analog scale of pain intensity. The duration of pain and frequency were also recorded. Patients were followed up at 1 week, 4 weeks, 3 months, 6 months, and 1 year and then at yearly intervals and monitored for recurrence of symptoms and corneal morphology. The removed epithelial sheet was examined by electron microscopy in 4 patients. Intervention The affected area of epithelium was peeled off after an application of 20% alcohol for 40 seconds under topical anesthesia. Eyes of patients were treated with an antibiotic and preservative-free artificial tear medication, and a bandage contact lens was inserted until epithelial healing was complete. Main Outcome Measures Frequency of recurrence of erosions and duration and intensity of symptoms after alcohol delamination were studied. Results Eleven of the 12 eyes of patients had dramatic relief of symptoms over the follow-up period, ranging from 6 to 40 months. Eight patients were symptom free, and 1 patient had 2 mild symptom episodes in the first posttreatment month before becoming symptom free. The average follow-up period was 23.5 months. There were no residual effects from the application of alcohol noted in any patient. Electron microscopy of removed epithelium showed features of the underlying pathology. The separation of the epithelium occurred at the interface of the subepithelial abnormal deposit and the surface of Bowman's zone. Conclusion Alcohol delamination appears to be a novel, simple, inexpensive treatment for RCEs. Unlike other methods, the removed epithelium is available as a sheet that may be subjected to further examination, though some of the changes observed may reflect the effect of alcohol on the epithelium.

PURPOSE. The incidence of acanthamoeba keratitis (AK) in the UK is some 15 times that in the United States and seven times that in Holland. To investigate reasons for this higher frequency, a study of the role of domestic tap water as a potential source of AK was undertaken. METHODS. Tap outlets from the homes of 27 patients with culture-proven AK were sampled and cultured for free-living amoebae (FLA). For all Acanthamoeba isolates, mitochondrial DNA (mtDNA) restriction fragment length polymorphisms (RFLPs) and cytochrome oxidase (cox 1/2) sequence typing was performed to determine the similarity between corneal and tap water isolates. RESULTS. FLA, including Acanthamoeba, were isolated from 24 (89%) of 27 homes, and the presence within the homes varied significantly with tap water temperature and location: 19 (76%) of 25 bathroom sink cold taps sampled compared with 6 (24%) of 25 hot and 9 (47%) of 19 kitchen cold taps compared with 3 (16%) of 19 of hot kitchen taps. Acanthamoeba were isolated from 8 (30%) of 27 homes (five bathroom sink cold taps, one cloakroom cold tap, one bath, and one bedroom sink mixer [hot/cold] taps). In six cases, identical Acanthamoeba mtDNA profiles were found for the clinical and home tap water isolates. In keeping with UK plumbing practice, 24 of 27 homes had internal roof water storage tanks to supply domestic taps, but the mains fed the kitchen cold tap. CONCLUSIONS. Water storage tanks promote colonization of domestic water with FLA, including Acanthamoeba, and hence increase the risk of AK. This accounts for the significantly greater incidence of AK in the UK and supports advice to avoid using tap water in contact lens care routines. (Invest Ophthalmol Vis Sci. 2004;45:165‐169) DOI:10.1167/iovs.03-0559

Purpose To determine the plane of cleavage of the corneal epithelium and smoothness of underlying stroma, after alcohol debridement in photorefractive keratectomy (PRK) and laser subepithelial keratectomy (LASEK). Methods The epithelial flap from six patients undergoing alcohol delamination of corneal epithelium before PRK and the epithelium and stroma from three eye bank donor eyes were fixed and processed for transmission (TEM) and scanning electron microscopy (SEM). The smoothness of the underlying stroma was studied by SEM and the plane of cleavage was determined by morphologic examination and morphometric measurements of basement membrane attached to the epithelial flap, using image-analysis software. Results A very smooth stromal bed, ideal for PRK was seen in the stroma of all three eye bank donor eyes after alcohol delamination. The plane of cleavage was determined to be at the hemidesmosomal attachments, including the most superficial part of the lamina lucida of the basement membrane. Conclusions Alcohol delamination of the corneal epithelium before PRK or LASEK consistently results in a very smooth cleavage at the level of the hemidesmosomal attachments, including the superficial lamina lucida. It leaves behind a very smooth surface, which is ideal for PRK. It also allows for an intact epithelial flap to be lifted as a sheet from the corneal surface and hence is ideally suited for the LASEK technique.

Actinomyces israelii are gram-positive filamentous bacteria forming yellow sulfur granules. They are the most well known complication of intrauterine contraceptive devices (IUCD). Healthcare staff dealing with reporting cervical smears should be aware of pseudoactinomyces entity in a cervical smears and biopsies as it may raise a false alarm to the clinician and may lead to unnecessary removal of IUCD and/or medical treatment.

BACKGROUNDThe basement membrane of human colonic mucosa contains numerous discrete pores. We have recently shown that following loss of the surface epithelium, many cells migrate out of the colonic lamina propria via basement membrane pores.AIMSTo characterise cells migrating out via basement membrane pores of the human gastric lamina propria, following loss of the surface epithelium.METHODSFresh human gastric mucosal samples were completely denuded of epithelial cells and placed in culture. Tissue samples were studied by electron microscopy (EM) and cells by EM, FACS analysis, immunohistochemistry, and reverse transcription polymerase chain reaction (RT-PCR).RESULTSEM showed numerous discrete pores (0.65–8.29 μm in diameter) in the subepithelial basement membrane. During culture of mucosal samples denuded of epithelial cells, lymphocytes, macrophages, and myofibroblasts migrated out of the lamina propria via the basement membrane pores. The lymphocytes were predominantly CD45RO+ and CD69+ T cells. Macrophages were shown to express cyclooxygenase (COX) 1 and 2 enzymes. Myofibroblasts were established in culture and, despite prolonged culture and passage, retained their phenotype. They expressed mRNA and protein for COX 1 and 2 enzymes and their release of prostaglandin E2 was inhibited by selective COX 1 and 2 inhibitors.CONCLUSIONSLamina propria cells migrating out of cultured denuded gastric mucosal samples have been characterised phenotypically and functionally. Such cells would be suitable for studies of their interactions with epithelial cells and also with Helicobacter pylori and its products.

We conducted a study to determine if preserved human amniotic membrane can reduce corneal haze induced by excimer laser photoablation. Excimer photoablation was performed bilaterally on 40 New Zealand white rabbits with a 6 mm ablation zone and 120 micrometer depth (PTK) using the VISX Star. One eye was randomly covered with a preserved human amniotic membrane and secured using four interrupted 10 - 0 nylon sutures; the other eye served as control. The amniotic membranes were removed at one week, and the corneal haze was graded with a slit-lamp biomicroscopy by three masked corneal specialists (WC, KH and RF) biweekly for the ensuing 12 weeks. Histology and in situ TUNEL staining (for fragmented DNA as an index for apoptosis) was performed at days 1, 3 and 7 and at 12 weeks. One week after excimer photoablation, the amniotic membrane-covered corneas showed more anterior stromal edema, which resolved at the second week. A consistent grading of organized reticular corneal haze was noted among the three masked observers. Such corneal haze peaked at the seventh week in both groups. The amniotic membrane-covered group showed statistically significant less corneal haze (0.50 plus or minus 0.15) than the control groups (1.25 plus or minus 0.35) (p less than 0.001). The amniotic membrane-covered corneas had less inflammatory response at days 1 and 3, showing nearly nil DNA fragmentation on keratocytes on the ablated anterior stromal and less stromal fibroblast activation. There is less altered epithelial cell morphology and less epithelial hyperplasia at 1 week in these amniotic membrane-treated eyes. We concluded from this study that amniotic membrane matrix is effective in reducing corneal haze induced by excimer photoablation in rabbits and may have clinical applications.

Editor,—Childhood uveitis is most frequently associated with juvenile chronic arthritis (JCA).1 The uveitis associated with JCA is anterior with no posterior involvement, although cystoid macular oedema may occur after cataract surgery. We describe disc neovascularisation (NVD) in two children with JCA associated chronic anterior uveitis. The possible aetiology of this and subsequent management are presented. ### CASE REPORTS #### Case 1 Left NVD was diagnosed in a white 13 year old girl with a 3 year history of bilateral anterior uveitis associated with pauciarticular onset JCA. There were no clinical or laboratory features to suggest any other disease process and she was completely asymptomatic apart from the involved joint. At presentation and throughout follow up she had a mild right anterior uveitis and moderate left anterior uveitis despite the constant use of topical steroids. No other form of immunomodulation had …

Objective To examine whether amniotic membrane transplantation (AMT), in preparing the perilimbal stroma, enhances the success of allograft limbal transplantation (ALT). Methods Thirty-one eyes of 26 consecutive patients had cytologically proven limbal deficiency resulting from chemical burns (14 eyes); Stevens-Johnson syndrome, toxic epidermal necrolysis, or pseudopemphigoid (5 eyes); contact lens–induced keratopathy (3 eyes); aniridia (3 eyes); multiple surgical procedures (2 eyes); atopy (2 eyes); or an unknown cause (2 eyes). Based on the severity of limbal deficiency, group A (mild), comprising 10 eyes, received AMT alone; group B (moderate), comprising 7 eyes, received AMT and ALT; and group C (severe), comprising 14 eyes, received AMT, ALT, and penetrating keratoplasty. All patients except those in group A received continuous oral cyclosporine. Results Except for the 2 eyes with atopy, all amniotic membrane–covered surfaces showed rapid epithelialization (in 2 to 4 weeks) and reduced inflammation, vascularization, and scarring, and the surfaces became smooth and wettable. For the mean follow-up period of 15.4 months, 25 (83%) of 30 eyes showed visual improvement, consisting of 6 or more lines (13 eyes), 4 to 5 lines (6 eyes), or 1 to 3 lines (6 eyes). Visual improvement decreased with the severity of limbal deficiency from 8 (100%) of 8 eyes in group A to 5 (71%) of 7 eyes in group B and 11 (79%) of 14 eyes in group C. In group C, corneal graft rejection occurred in 9 (64%) of 14 eyes, and reversible early limbal allograft rejection was noted in 3 (14%) of 21 eyes of groups B and C. Conclusions For partial limbal deficiency with superficial involvement, AMT alone is sufficient and hence superior to ALT because there is no need to administer cyclosporine. For total limbal deficiency, additional ALT is needed, and AMT helps reconstruct the perilimbal stroma, with reduced inflammation and vascularization, which collectively may enhance the success of ALT.

Interactions between epithelial cells and subepithelial myofibroblasts are increasingly recognized as important in the regulation of epithelial cell function. We have established primary cultures of subepithelial myofibroblasts from adult human colonic mucosal samples denuded of epithelial cells and maintained in culture. During culture of mucosal tissue, subepithelial myofibroblasts migrated out via basement membrane pores before establishment in culture. Despite prolonged culture and passage, the myofibroblasts maintained their phenotype, as demonstrated by expression of α-smooth muscle actin and vimentin. The cells expressed transcripts and protein for cyclooxygenase (COX)-1 and -2 enzymes, and their release of prostaglandin E2(PGE2) was inhibited by selective COX-1 and -2 inhibitors. The myofibroblasts also expressed the extracellular matrix (ECM) proteins collagen type IV, laminin-β1and -γ1, and fibronectin. Adult human colonic subepithelial myofibroblasts may influence epithelial cell function via products of COX-1 and -2 enzymes, such as PGE2and secreted ECM proteins.

SUMMARY Lymphocytes and macrophages are present in the normal intestinal lamina propria, separated from the epithelial monolayer by the basement membrane. There is evidence for movement of mononuclear cells through the lamina propria, entering from the systemic circulation and exiting via lymphatic channels. The goal of our studies was to investigate the capacity of cells to migrate out from the lamina propria into the lumen following the loss of surface epithelial cells. An in vitro model was therefore established in which normal human intestinal mucosal samples, denuded of the surface epithelium, were maintained in culture. Electron microscopy showed that during culture, large numbers (> 2 × 106/g tissue per 24 h) of cells migrated out of the lamina propria via discrete ‘tunnels’ which were in continuity with pores (diameter

We have studied the brains of 10 patients with clinically and pathologically defined Huntington's disease and graded the degree of striatal pathology according to the Vonsattel grading system. Sections from nine cerebral cortical areas (Brodmann areas 8. 10, 24, 33, 28, 38, 7, 39, 18), the cerebellum, hypothalamus, medulla and caudate nucleus were stained with antibodies to ubiquitin and ubiquitin C-terminal hydrolase (PGP 9.5). Dystrophic neurites. immunoreactive with ubiquitin and PGP 9.5 were detected in all cortical areas. in layers 3, 5 and 6, of all brains studied. No dystrophic neurites were found in subcortical areas or cerebellum. Sections from cortical areas 8 and 24 from the two brains with the most and least ubiquitin-immunoreactive neurites were stained with antibodies to β-amyloid precursor protein, tau, glial fibrillary acidic protein, neurofilament protein, αB crystallin, GABA, cholecystokinin and somatostatin. The dystrophic neurites were found to also react with β-amyloid precursor protein. Electron microscopy showed the abnormal neurites to contain granulo-filamentous material. Granular deposits with a diameter of 40–100 nm were interspersed between randomly orientated ‘fuzzy’ or coated, straight or slightlv curved filaments measuring 10–15 nm in diameter. These structures have not been seen in control brain and differ from age-related neuritic degeneration and neurites associated with amyloid. Immunohistochemically these structures most resemble CA 2/3 neurites seen in Lewy body disease, and, ultrastructurally, the intraneuronal filamentous inclusions in motor neuron disease. The areal density of these neurites was quantified in 20 microscopic fields in the superior frontal and anterior cingulate sections (Brodmann areas 8 and 24) and did not correlate with the Vonsattel grade, suggesting that they are an independent and possibly primary cortical pathology in Huntington's disease.

Qualitative histological studies in the distribution of urokinase-type plasminogen activator (uPA) in human colorectal carcinomas have been well documented. However, to our knowledge the histological distribution of this enzyme has not been quantified in any tumour. For the present image analysis study, uPA was demonstrated in sections of human colorectal cancer using immunoperoxidase technique. A total of 9 colorectal carcinoma cases were used, in which 132 regions were analysed. Within each region, staining intensity measurements were made at evenly spaced intervals. Samples of normal mucosa from 6 cases were also studied. Enzyme levels were assessed with staining intensity measurements. For each section, a negative control section was included, in which the primary antibody was omitted. Staining for uPA was quantified for each region in the test section, and the measurement for the corresponding region of the negative control was then subtracted. The enzyme uPA was localised more at the tumour edge than in the tumour centre or distant host tissue. These differences were highly significant (p < 0.0001). There was also a highly significant difference in staining intensity when tumour regions adjacent to pushing edge were compared with those adjacent to infiltrating edge (p < 0.0001). Infiltrating tumours showed stronger staining for uPA than tumours with pushing edges. Since invasive activity is thought to be maximal at the edge of the tumour, localisation of uPA at this site is consistent with the role of this enzyme in the process of tumour invasion. © 1995 Wiley-Liss, Inc.

Previous studies on colorectal carcinomas indicate that consistent differences in epithelial basement membrane (EBM) integrity are present between the tumour centre and periphery. We report that within the tumour centre, EBM staining between back-to-back (BTB) neoplastic glands (i.e., adjacent glands in direct contact with no intervening connective tissue) generally follows a pattern different from that of EBM staining at the tumour:stromal interface (TSI). Such distinctions are important, since the factors responsible for EBM deficiencies may vary with intra-tumoural location, as may the prognostic significance of these deficiencies. Analysis of paraffin sections from 130 colorectal carcinoma cases showed that EBM staining between BTB glands is generally weaker and more discontinuous than at the TSI, sometimes appearing as a linear array of immunostained granules on high-resolution light microscopy. By double-labelling immunofluorescence analysis of cryostat sections from 30 cases, a decrease in type-IV collagen:laminin staining intensity ratio was found between BTB glands. Hence, the composition of EBM between BTB glands appears to be abnormal. As much recent evidence indicates that epithelial:mesenchymal interactions play an essential role in EBM formation, the demonstration of immunostained EBM fragments between BTB glands requires an explanation: We suggest that the synthesis of EBM between BTB glands involved previously intervening stromal (mesenchymal) cells, and that EBM fusion and dissolution occur between BTB glands following the displacement of these cells.

Continuous or intermittent consumption by rats of food moulded by Penicillium aurantiogriseum induced prominent and extensive histopathological changes within several weeks seen specifically at the renal cortico-medullary junction. Many cells of the P3 segment of proximal tubules contained either giant nuclei or multiple enlarged nuclei, described in this text as karyomegaly, but also included within a cytomegalic change. The changes contrasted with the tubular cell necrosis and concomitant mitosis elicited after only four days consumption of nephrotoxic mould. Unilateral nephrectomy enabled persistence of histopathological changes to be assessed directly after detailed histology at an earlier stage. After ten days consumption of food with a 100-fold excess of fungal extract containing the amphoteric nephrotoxins, the typical acute histopathology evolved, over a period of three weeks on normal diet, into the bizarre karyomegalic histopathology, implying a latent effect. Karyomegaly persisted for at least twelve months after nephrotoxin dosage ceased. P. aurantiogriseum karyomegaly was much more striking than that induced by a relatively high chronic dose of another Penicillium nephrotoxin, ochratoxin A. Although the study does not attempt to measure relative potencies, qualitatively similar ultrastructural changes (enlarged nuclei, proliferation of smooth endoplasmic reticulum and thickening of proximal tubule basement membranes) were induced by the two types of nephrotoxin. The broadly toxic ochratoxin A is the popular putative aetiological agent in the mysterious and insidious Balkan endemic nephropathy and associated urinary tract tumours. As the renal carcinogenicity of ochratoxin A in rats follows karyomegaly, the striking karyomegaly induced by P. aurantiogriseum in the proximal tubules of the kidney must be considered as a potential factor in human chronic renal disease.