Failure of ES-62 to inhibit T-helper type 1 responses to other filarial nematode antigens.

ES-62 is a secreted protein of filarial nematodes that possesses multiple immunomodulatory activities. A full characterization of these activities awaits elucidation but to date it has been shown that ES-62 can inhibit pro-inflammatory/Th1 immune responses and in some studies, it has been found to actively support Th2 development. As an active filarial nematode infection is associated with a Th2-like immunological phenotype, this study investigated whether ES-62 was likely to be responsible for, or at least contribute to, this phenotype. Specifically, we determined ES-62's effect on the immune response to two other filarial nematode antigens, chosen for their ability to promote Th1 responses. The two antigens were recombinant Onchocerca volvulus-Fatty acid And Retinol-binding-1 (rOv-FAR-1) and recombinant Onchocerca volvulus-Activation associated Secreted Protein-1 (Ov-ASP-1). Overall the results show that in spite of its previously characterized immunomodulatory properties, ES-62 was unable to modulate/reverse the Th1 immune responses induced by the two Onchocerca antigens. Therefore, in this study no support is provided for the idea that ES-62 might be a major player in facilitating the overall immunological phenotype in filariasis and reasons for this somewhat surprising outcome are discussed.

Does delay in referral of proliferative diabetic retinopathy from the diabetic eye screening programme lead to visual loss?

AimsTo ascertain the effect on visual acuity (VA) of a delay in Hospital Eye Service (HES) consultation for patients referred with proliferative diabetic retinopathy (PDR; R3) from the Diabetic Eye Screening Programme (DESP).MethodsAll patients referred to Moorfields Eye Hospital from DESP between April and December 2013 with a referral diagnosis of PDR in at least one eye were eligible. Screening programme VA was compared with VA at first HES appointment and final follow-up appointment. Reasons for any VA loss were noted.ResultsA total of 86 patients were included. Of these, 28 (33%) were seen in more than 4 weeks after their DESP referral. At first HES appointment, 39 (45%) patients were graded as having active PDR in at least one eye. Delay in referral did not significantly predict the likelihood of vision loss in all patients referred (χ(2), P=0.49) or in just those patients with a definitive HES diagnosis of active PDR (χ(2), P=1.00). In only 3 patients with active PDR was a delay in presentation thought to have led directly to VA loss.ConclusionsThere may be minimal short-term visual consequence in several weeks of delayed referral for many patients with a diagnosis of R3. However, the national guidance remains important. This is due to the occasional patient at very high risk of vision loss and the many gains for the patients in terms of time to properly assess medical and ocular conditions and counsel and consent them for treatment where necessary.

Structural changes of the choroid in sarcoid- and tuberculosis-related granulomatous uveitis.

AIM: The aim of this study is to characterise the choroidal features of patients diagnosed with sarcoid- and tuberculosis (TB)-associated granulomatous uveitis using spectral domain optical coherence tomography (OCT). METHODS: Twenty-seven patients (27 eyes) diagnosed with sarcoid- (13 eyes) and TB (14 eyes)-related uveitis were included in this retrospective, cross-sectional study. Over a six-month period, patients diagnosed with sarcoid and TB granulomatous uveitis were scanned using enhanced depth imaging OCT. Clinical and demographical characteristics were recorded, including the method of diagnosis, disease activity, site of inflammation (anterior or posterior), treatments, and visual acuity (VA). Manual segmentation of the choroidal layers was performed using custom image analysis software. RESULTS: The main outcome measure was OCT-derived thickness measurements of the choroid and choroidal sublayers (Haller's large vessel and Sattler's medium vessel layers) at the macula region. The ratio of Haller's large vessel to Sattler's medium vessel layer was significantly different at the total macula circle in eyes diagnosed with TB uveitis (1.47 (=140.71/95.72 µm)) compared with sarcoid uveitis (1.07 (=137.70/128.69 µm)) (P=0.001). A thinner choroid was observed in eyes with a VA ≥0.3 LogMAR (Snellen 6/12; 198.1 µm (interquartile range (IQR)=147.0-253.4 µm) compared with those with VA <0.3 LogMAR (292.4 µm (IQR=240.1-347.6 µm)) at the total macula circle (P=0.004). At the foveal central subfield, the median choroidal thickness was 336.8 µm (IQR=272.3-375.4 µm) in active compared with 239.3 µm (IQR=195.3-330.9 µm) in quiescent disease (P=0.04). CONCLUSION: A disproportionately enlarged Sattler's layer may indicate a diagnosis of sarcoid-related uveitis, and choroidal thickening may be a feature of active granulomatous uveitis.

Bullous pemphigoid sera that contain antibodies to BPAg2 also contain antibodies to LABD97 that recognize epitopes distal to the NC16A domain.

IgG antibodies from the sera of some patients with bullous pemphigoid (BP) react with a 180 kDa protein termed BPAg2. Antibodies in BP are directed to an extracellular noncollagenous domain of this protein termed NC16A. Our group has recently shown that a portion of the extracellular domain of BPAg2 is identical to LABD97 on the basis of amino acid sequencing. We evaluated sera from 33 patients with BP with circulating IgG antibodies on indirect immunofluorescence, which stained the epidermal side of split skin with titers ranging from 1:40 to 1:640. Immunoblotting was performed against (i) two preparations of proteins from epidermal extract, one containing BPAg2 and one containing LABD97, and (ii) the recombinant NC16A domain of the BPAg2 protein. Twelve sera reacted with the BPAg2 protein. Ten of these also reacted strongly with the NC16A domain. Nine of the 12 sera also reacted with the LABD97 antigen. Bound antibodies were eluted from the 97 kDa band and reapplied to split skin where they bound to the epidermal side. The eluted antibodies also reacted to the BPAg2 protein from the epidermal extract, but did not react with the NC16A domain on immunoblot. We conclude that these nine sera react with an epitope present within BPAg2 and LABD97 but not within the NC16A domain. This epitope is therefore distal to the previously described epitopes in BP. In BP, epitope spreading may occur and antibodies may be produced that recognize the distal portion of the BPAg2 molecule identical to LABD97 but that do not involve the NC16A domain.

Analysis of telomere lengths in human corneal endothelial cells from donors of different ages.

PURPOSE: To investigate the telomere hypothesis of cellular aging as the mechanism for cell cycle arrest in normal human corneal endothelium. METHODS: The corneal endothelium and epithelium from 21 human corneas from 13 donors 5 weeks to 84 years of age were dissected and frozen at -70 degrees C. Purified DNA, digested with the restriction enzyme, HinfI, was run on 0.7% agarose gels, probed with radiolabeled (AATCCC)4, and exposed to a phosphor screen. The length of the terminal restriction fragment (TRF) was determined by densitometry. RESULTS: The cells of the corneal endothelium had TRF lengths ranging from 11.0 to 14.0 kbp (mean, 12.2 +/- 0.9). Corneal epithelial specimens showed TRF lengths that were always less than (mean, 10.4 +/- 1.0; range 9.0-12.0) the corresponding endothelial TRF lengths. Human corneal endothelial cells, transformed with human papillomavirus type 16 oncogenes E6 and E7, showed decreasing TRF lengths from 11 kbp at population doubling level (PDL) 15 to 9.5 kbp at PDL 73. Neither the endothelial and epithelial cells from human donors nor the transformed pre-immortalized human endothelial cells showed evidence of telomerase activity. CONCLUSIONS: Human corneal endothelial cells have long telomeres throughout life. Their limited replicative ability does not appear to result from critically short telomere lengths.

Effect of dorzolamide on corneal endothelial function in normal human eyes.

PURPOSE: To assess the effects of dorzolamide hydrochloride, a topical carbonic anhydrase inhibitor, on corneal endothelial function. METHODS: The authors measured the rate of corneal deswelling and the endothelial permeability to fluorescein after 2 hours of hypoxic contact lens wear in 19 normal human subjects. The study was double-masked; one eye of each subject was randomly assigned to receive 2% dorzolamide drops, and the other eye received placebo drops every 8 hours for 24 hours before the study day and twice during the study day. RESULTS: Dorzolamide-treated eyes were not significantly different from placebo-treated eyes in corneal deswelling rate, expressed as the percent recovery per hour (55.7% +/- 13.6% versus 59.6% +/- 14.5%; P > or = 0.10), open eye steady state thickness, swelling induced by hypoxia, and corneal autofluorescence. Endothelial permeability to fluorescein was increased in the dorzolamide eyes (4.40 +/- 0.84 x 10(-4) cm/minute versus 4.10 +/- 0.80 x 10(-4) cm/minute; P = 0.01). As expected, the intraocular pressure and aqueous humor flow rate were decreased in the dorzolamide eyes. CONCLUSIONS: Dorzolamide hydrochloride, when topically administered to normal human eyes for 24 hours, had no significant effect on the corneal deswelling rate after hypoxic stress. The corneal endothelial permeability to fluorescein, however, was increased by the drug, although this did not result in increased corneal thickness.

Vulvar melanoma in childhood.

BACKGROUND: Malignant melanoma is rarely diagnosed during childhood. Approximately 2% of malignant melanomas occur in patients younger than 20 years, with 0.3% to 0.5% of cases occurring in prepubescent children. In adult females malignant melanoma of the vulva and vagina is 100-fold less common than malignant melanoma of nongenital skin. Malignant melanoma of the vulva occurring in a child has been reported once before. OBSERVATIONS: We report 2 cases of childhood vulvar malignant melanoma presenting in preteenage girls. In both cases, the lesions were asymptomatic enlarging hyperpigmented macules on the labium minus. In addition to features diagnostic of malignant melanoma, histological evidence of lichen sclerosus et atrophicus was identified in both lesions. Local excision with conservative margins was the treatment modality of choice in both cases, with good preservation of anatomic structure and function. CONCLUSIONS: This report is of 2 cases of vulvar melanoma in childhood, a rare, yet potentially devastating, presentation of melanoma. Biopsies on suspicious pigmented lesions on the vulva of prepubescent children should be done to rule out malignant change.

An atypical phenotype of macular and peripapillary retinal atrophy caused by a mutation in the RP2 gene.

AIMS: To determine the molecular basis and describe the phenotype of an atypical retinal dystrophy in a family presenting with bilateral, progressive central visual loss. METHODS: Family members were examined. Investigations included Goldman perimetry, electrophysiology, and autofluorescence imaging. Candidate gene screening was performed using SSCP and sequence analysis. The proband's lymphoblastoid cells were examined for protein expression. RESULTS: Fundal examination of the proband, his mother, and brother revealed peripapillary and macular atrophy. Autosomal dominant retinal dystrophy was suspected, but less severe disease in the mother led to screening for mutations in X linked genes. A 4 bp microdeletion in exon 3 of the RP2 gene, segregating with disease, was identified. No RP2 protein expression was detected. CONCLUSION: The distinct phenotype in this family, caused by this frameshifting mutation in RP2, broadens the phenotypic spectrum of X linked retinitis pigmentosa. The absence of RP2 protein suggests that loss of protein function and not novel gain of function could account for the atypical phenotype. A definitive diagnosis of X linked retinitis pigmentosa permits appropriate genetic counselling with important implications for other family members. Clinicians should have a low threshold for screening RP2 in families with retinal dystrophy, including posterior retinal disease, not immediately suggestive of X linked inheritance.

Prospective study of the SI-40NB foldable silicone intraocular lens.

PURPOSE: To evaluate the surgical performance and postoperative outcomes of 100 consecutive implantations of the new AMO PhacoFlex SI-40NB foldable intraocular lens (IOL). SETTING: Concord Hospital and Dalcross Private Hospital, Sydney, Australia. METHODS: This prospective study comprised 100 consecutive patients who had endocapsular phacoemulsification cataract surgery with implantation of the AMO SI-40NB IOL, which has a silicone optic and poly(methyl methacrylate) (PMMA) haptics. Patients were excluded if trabeculectomy was performed concomitantly. The main outcome measures were control of folding, ease of implantation, centration, intraoperative and postoperative complications, and visual acuity. A minimum of 4 months follow-up was planned. RESULTS: Mean follow-up was 4.3 months +/- 2.1 (SD) (range 1 to 12 months). The lens permitted excellent control of folding and insertion, as well as excellent centration at surgery and 4 months. There was one lens-related intraoperative complication because of crimping of the trailing lens haptic. Postoperative complications were minimal, and 92% of patients achieved a visual acuity of 20/20 or better at 1 and 4 months. Excluding the eyes with retinal pathology, 100% achieved 20/20 or better at 4 months. CONCLUSION: The AMO SI-40NB three-piece silicone IOL with PMMA haptics provided excellent operative control, centration, and visual outcome and is suitable for small incision cataract surgery.

The clinical and immunopathological manifestations of anti-epiligrin cicatricial pemphigoid, a recently defined subepithelial autoimmune blistering disease.

Cicatricial pemphigoid (CP) is a rare, acquired, autoimmune, subepithelial blistering disease. It primarily affects mucous membranes but it also may involve the skin. Morbidity is associated with the propensity for scar formation and may be especially severe when mucosal surfaces such as the conjunctivae, larynx, esophagus, or urethra are involved. On direct immunofluorescence microscopy, CP is characterized by the linear deposition of immunoreactants, principally IgG and C3, along epithelial basement membranes. Over the last 10 years, studies in a number of laboratories have shown that circulating autoantibodies in patients with CP may target one of several different autoantigens. One subset of patients with the CP-phenotype have IgG anti-basement membrane autoantibodies against laminin 5 (alpha3beta3gamma2) (i.e., patients with anti-epiligrin CP [AECP]). This review discusses recent advances in the understanding of CP and emphasizes salient features of AECP pathophysiology.

Dermatitis Herpetiformis: a review of fifty-four patients.

A review of 54 patients with dermatitis herpetiformis presenting between 1984-1993 to The Regional Centre of Dermatology, Mater Misericordiae Hospital was undertaken. All patients had skin lesions clinically and histologically typical of dermatitis herpetiformis. Deposition of granular IgA at the dermoepidermal junction on direct immunofluorescence was present in each case. The average age of onset was 41.8 yr, patients having symptoms for an average of 1.6 yr before diagnosis. Six patients had a prior history of coeliac disease. Two patients had a family history of dermatitis herpetiformis, a father and son who were both propositi in this study. Small bowel biopsy was performed on 35 patients, 71.4 per cent of them showing evidence of villous atrophy. All patients were controlled on a gluten free diet or by dapsone or a combination of these. None of the patients experienced serious adverse effects of therapy, nor did any develop lymphoma of the small bowel with a mean follow up period of 4.2 yr (range 1-10 yr).

A detailed phenotypic description of autosomal dominant cone dystrophy due to a de novo mutation in the GUCY2D gene.

PURPOSE: The purpose of this study is to describe the phenotype of a family with de novo mutation in the GUCY2D. MATERIALS AND METHODS: Five subjects, including two monozygotic twins, underwent ophthalmic clinical examination while some had autofluorescence imaging (AF) and optical coherence tomography (OCT). Symptomatic individuals underwent electrophysiological testing. The youngest subject (21 years) was also evaluated psychophysically. DNA obtained from the individuals was screened for mutations in GUCY2D. Microsatellite markers were used to determine the haplotype of 17p surrounding the GUCY2D gene. RESULTS: The youngest subject had 6/18 visual acuity, an annulus of hyper-autofluorescence in the perifoveal region, and a subfoveal absence of outer segments on OCT. In the older individuals, severe thinning of inner retina and a patchy loss of photoreceptors and retinal pigment epithelium were observed in the perifoveal region. All three showed generalised cone system dysfunction with preserved rod function on electrophysiology. Psychophysical evaluation was consistent with poor cone function. Screening of the GUCY2D gene revealed the mutation p.R838H in all the affected individuals and was absent in the asymptomatic patients. Haplotyping showed that the mutation originated from the unaffected mother. CONCLUSIONS: Autosomal dominant cone dystrophy due to GUCY2D can occur without a history in the antecedents due to a de novo mutation. This is important to consider in any simplex case with a similar phenotype. The phenotype description of this disorder is expanded with detailed description of the OCT findings. This paper describes the concordance of the phenotypic findings in the monozygotic twins.