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Diabetic retinopathy is a leading cause of blindness. The purpose of this study is to identify novel genetic loci associated with the sight threatening complications of diabetic retinopathy. We performed a meta-analysis of genome-wide association data for severe diabetic retinopathy as defined by diabetic macular edema or proliferative diabetic retinopathy in unrelated cases ascertained from two large, type I diabetic cohorts: the Genetics of Kidney in Diabetes (GoKinD) and the Epidemiology of Diabetes Intervention and Control Trial (EDIC) studies. Controls were other diabetic subjects in the cohort. A combined total of 2829 subjects (973 cases, 1856 controls) were studied on 2 543 887 single nucleotide polymorphisms (SNPs). Subjects with nephropathy were excluded in a sub-analysis of 281 severe retinopathy cases. We also performed an association analysis of 1390 copy number variations (CNVs) using tag SNPs. No associations were significant at a genome-wide level after correcting for multiple measures. The meta-analysis did identify several associations that can be pursued in future replication studies, including an intergenic SNP, rs476141, on chromosome 1 (P-value 1.2 × 10(-7)). The most interesting signal from the CNV analysis came from the sub-group analysis without nephropathy subjects and is rs10521145 (P-value 3.4 × 10(-6)) in the intron of CCDC101, a histone acetyltransferase. This SNP tags the copy number region CNVR6685.1 on chromosome 16 at 28.5 Mb, a gain/loss site. In summary, this study nominates several novel genetic loci associated with the sight-threatening complications of diabetic retinopathy and anticipates future large-scale consortium-based validation studies.

Cutaneous aspergillosis is a rare infection most often seen in immunocompromised patients. We report a case of primary cutaneous aspergillosis infection in a nonhealing scalp wound of an immunocompetent elderly patient. The patient had a cutaneous malignancy of the scalp treated with surgical excision but complicated by poor wound healing. Fungal culture of the nonhealing wound revealed Aspergillus niger. The nonhealing wound subsequently resolved with retapamulin ointment 1% and ketoconazole gel 2%.

Purpose: The purpose of this study is to report a case of acute zonal occult outer retinopathy and to describe a novel method of imaging photoreceptor damage using Cirrus spectral-domain optical coherence tomography (SD-OCT; Cirrus, Carl Zeiss Meditec, Dublin, CA) en face visualization. Our examination also included color fundus photographs, Goldmann visual field, fluorescein and indocyanine green angiography, fundus autofluo-rescence, and Ganzfeld electroretinography., Methods: This is a case report of a single patient., Results: Despite 20/20 visual acuity in both of our patient's eyes, Goldmann visual field showed a unilateral C-shaped scotoma centered on the left optic nerve, sparing the fovea. Fundus and angiographic findings showed nonspecific pigmentary changes and hyperfluo-rescent foci along the superior and inferior vascular arcades. The electroretinography showed decreased scotopic amplitudes, with impaired 30-Hz flicker response, and Cirrus SD-OCT showed loss of the inner segment/outer segment photoreceptor junction. Cirrus SD-OCT Advanced Visualization three-dimensional reconstruction with en face visualization delineated an area of inner segment/outer segment attenuation corresponding to the Goldmann visual field scotoma. The patient's condition was unchanged at 5 months follow-up., Conclusion: In addition to classic diagnostic modalities, photoreceptor attenuation observed with Cirrus SD-OCT can contribute to the diagnosis of acute zonal occult outer retinopathy and may help elucidate the etiology of this puzzling condition.

Listeria monocytogenes can cause severe invasive disease in humans and has been isolated from a variety of foods. This study aimed to investigate type diversity and distribution across sources by subtyping via PFGE a set of 300 L. monocytogenes isolates collected in Italy from foods over a five year period (from 2003 to 2007). The most frequent serotypes were 1/2a (45%), 1/2c (22%), and 4b/4e (16%); 5% of the isolates were untypeable by conventional serotyping. Significant associations were observed between serotype 1/2a with dairy (O.R.=13.9) and 1/2c with meat (O.R.=33.3). All isolates were typeable, generating 164 combined PFGE profiles. Of these, 121 were unique, being displayed by only one isolate. The other 43 profiles grouped the remaining isolates and were shared between two (N=22), three (N=10), four (N=3) and five isolates (N=4). The remaining 4 profiles were shared between 7, 14, 17 and 46 isolates, respectively. Some profiles (N=7) were retrieved in samples collected in different years, indicating persistency in foods and processing plants. This research may pose the ground for designing a broad typing database which could ease the understanding of L. monocytogenes diversity and could be used for facilitating epidemiological investigations for the identification of listeriosis outbreaks. Data show how large subtype databases may facilitate the identification of common and source-specific types. More comprehensive databases may be needed to fully understand L. monocytogenes diversity and to provide useful data to be considered in epidemiological investigations., (Copyright 2010 Elsevier B.V. All rights reserved.)

Purpose: To describe a case of sarcoidosis manifesting as a unilateral posterior uveitis and retinal arteritis., Design: Interventional case report., Methods: Clinical examination, fluorescein angiography, radiological imaging, and laboratory testing revealed evidence of a primary arteritis associated with sarcoidosis., Results: A 39-year-old white male presented with unilateral decreased vision, disc edema, exudative maculopathy, and retinal arteritis. Lab work and imaging were consistent with sarcoidosis. The patient responded to a 2-month course of oral prednisone and a sub-Tenon's triamcinolone injection., Conclusions: Sarcoidosis should be considered in the differential diagnosis of uveitis associated with a primary retinal arteritis.

Mutations in genes associated with the U4/U6-U5 small nuclear ribonucleoprotein (snRNP) complex of the spliceosome are implicated in autosomal-dominant retinitis pigmentosa (adRP), a group of progressive retinal degenerative disorders leading to visual impairment, loss of visual field, and even blindness. We recently assigned a locus (RP33) for adRP to 2cen-q12.1, a region that harbors the SNRNP200 gene encoding hBrr2, another U4/U6-U5 snRNP component that is required for unwinding of U4/U6 snRNAs during spliceosome activation and for disassembly of the spliceosome. Here, we report the identification of a missense mutation, c.3260C>T (p.S1087L), in exon 25 of the SNRNP200 gene in an RP33-linked family. The c.3260C>T substitution showed complete cosegregation with the retinitis pigmentosa (RP) phenotype over four generations, but was absent in a panel of 400 controls. The p.S1087L mutation and p.R1090L, another adRP-associated allele, reside in the "ratchet" helix of the first of two Sec63 domains implicated in the directionality and processivity of nucleic acid unwinding. Indeed, marked defects in U4/U6 unwinding, but not U4/U6-U5 snRNP assembly, were observed in budding yeast for the analogous mutations (N1104L and R1107L) of the corresponding Brr2p residues. The linkage of hBrr2 to adRP suggests that the mechanism of pathogenesis for splicing-factor-related RP may fundamentally derive from a defect in hBrr2-dependent RNA unwinding and a consequent defect in spliceosome activation.

In this study we investigated the microbiota of Gorgonzola rinds and maturing shelf swabs collected in 5 different maturing cellars in the Northwest part of Italy, in association with the detection and characterization of Listeria monocytogenes. Culture-dependent and -independent methods were performed in order to profile the main microbial populations present on the rinds and in the maturing shelves and species-specific PCR and Pulsed Field Gel Electrophoresis (PFGE) were used to identify and type L. monocytogenes isolates. The microflora was predominated by lactic acid bacteria and coagulase negative cocci, while enterococci and yeasts were very variable between the samples. Arthrobacter sp., Carnobacterium sp., Staphylococcus sp. and Brevibacterium linens, as bacteria, and Debaryomyces hansenii, as yeast, were detected by Denaturing Gradient Gel Electrophoresis (DGGE). Cluster analysis of the DGGE profiles clearly highlighted a cellar-specific microflora. L. monocytogenes was isolated in 11.1% of the rinds and 29.4% of the swabs and the molecular characterization of the isolates suggests a route of contamination from the maturing shelves to the rinds. No correlation was found between DGGE profiles and presence or absence of L. monocytogenes.

Salmonella is one of the most common causes of human gastroenteritis often associated with pork consumption. The aims of this cross-sectional study were to collect preliminary data on the presence of Salmonella enterica in pigs in Piedmont (Italy), through sampling on farm and at slaughter and to gather pilot data on serotypes and phagetypes present in the sampled area and distribution of anti-microbial resistance among isolated strains. Salmonella was detected through culture and identified with Salmonella spp. and Salmonella Typhimurium PCR; positive samples were serotyped, phagetyped and tested for antibiotic susceptibility. Positive samples (from 9% of faeces up to 29% of tonsils) were found in 64% of the herds. Salmonella spp. was retrieved also from scalding water. Most of the isolates were Salmonella Derby, Salmonella Typhimurium and Salmonella 4,5,12:i:-. The results of Salmonella Typhimurium specific PCR suggested that Salmonella 4,5,12:i:- might be unrecognized by serotyping. Anti-microbial resistance was recorded in 75-100% of the isolates. Phagetyping allowed the identification of DT104B and DT46A strains. These results set the bases for further research studies that would aim to estimate the real herd prevalence in Piedmont and the diffusion of serotypes and anti-microbial resistant strains within the same region.

Purpose: To determine whether patient self-report of prior laser treatment can be used as a reliable tool for assessing the presence of severe diabetic retinopathy., Design: This was a retrospective study on two groups of diabetic subjects., Methods: One hundred patients with diabetes were recruited from the general eye and retina clinics at the University of Chicago Hospitals. The patients were asked, "Have you ever received laser treatment for your diabetic eye disease (DED)?" A chart review was then conducted noting if the patient had received either focal laser treatment for diabetic macular edema or panretinal photocoagulation for proliferative diabetic retinopathy. Data from the Wisconsin Epidemiological Study of Diabetic Retinopathy (WESDR) were also analyzed. Participant responses to the question "Have you had laser photocoagulation treatment for your eyes?" were analyzed with documentation of photocoagulation scars determined by grading seven-standard field color fundus photographs., Results: In the University of Chicago group, 96 of 100 (96%) of patients were accurate in reporting whether they had received previous laser treatment for DED (sensitivity 95.8%, specificity 96.1%, and positive predictive value 88.5%). In the WESDR analysis, 2,329 of 2,348 (99%) of participants were accurate in reporting whether they had prior laser treatment for DED (sensitivity 96.0%, specificity 99.5%, and positive predictive value 95.6%)., Conclusions: The high sensitivity and specificity of our results validate the use of patient self-report as a useful tool in assessing past laser treatment for severe diabetic retinopathy. Patient self-report may be a useful surrogate to clinical examination or medical record review to determine the presence of severe diabetic retinopathy.

Purpose: To use the power of knowledge acquisition and machine learning in the development of a collaborative computer classification system based on the features of age-related macular degeneration (AMD)., Methods: A vocabulary was acquired from four AMD experts who examined 100 ophthalmoscopic images. The vocabulary was analyzed, hierarchically structured, and incorporated into a collaborative computer classification system called IDOCS. Using this system, three of the experts examined images from a second set of digital images compiled from more than 1000 patients with AMD. Images were annotated, and features were identified and defined. Decision trees, a machine learning method, were trained on the data collected and used to extract patterns. Interrelationships between the data from the different clinicians were investigated., Results: Six drusen classes in the structured vocabulary were largely sufficient to describe all the identified features. The decision trees classified the data with 76.86% to 88.5% accuracy and distilled patterns in the form of hierarchical trees composed of 5 to 15 nodes. Experts were largely consistent in their characterization of soft, and to a lesser extent, hard drusen, but diverge in definition of other drusen. Size and crystalline morphology were the main determinants of drusen type across all experts., Conclusions: Machine learning is a powerful tool for the characterization of disease phenotypes. The creation of a defined feature set for AMD will facilitate the development of an IDOCS-based classification system.

Objective: To identify the genetic factors associated with familial non-arteritic anterior ischemic optic neuropathy (NA-AION) in a large pedigree., Methods: Eleven family members of a single pedigree, including six affected with NA-AION, underwent detailed clinical examinations. The mitochondrial DNA of the proband was sequenced in its entirety in search of disease-causing mutations associated with NA-AION in the pedigree. A control panel comprising 1488 patients suspected of having Leber hereditary optic neuropathy (LHON) and 97 general-population control subjects was screened for the mitochondrial sequence variant identified in the family., Results: Affected family members were all male and exhibited classic features of NA-AION. Their mean age was 50.2 +/- 5.0 years. A total of 23 sequence variations were detected in the mitochondrial genome of the proband, including one novel sequence variation (G4132A, Ala276Thr) in the NADH dehydrogenase subunit 1 gene (ND1). The G4132A mitochondrial variant was detected in six members of a single pedigree with NA-AION. The G4132A variation was not observed in any of the 1585 subjects in the control panel. Moreover, this variant was not identified in over 2469 ethnically diverse individuals previously evaluated through the Human Mitochondrial Genome Database. None of the three major mutations associated with LHON (G3460A, G11778A, T14484C) were identified in the family., Conclusion: The G4132A mitochondrial variation is associated with familial NA-AION in our pedigree.

Objective: To determine the histidine frequency in patients with the cuticular drusen phenotype of age-related macular degeneration (AMD)., Methods: Fifty individuals were identified who met the criteria for the cuticular drusen phenotype using a standard threshold photograph. We genotyped DNA samples using a polymerase chain reaction-based restriction digest assay. Seven hundred individuals with typical AMD and 252 controls were also genotyped. Fisher exact test was used to analyze the significance of allele frequency differences., Results: The histidine variant was present in 70% (frequency +/- SE, 0.70 +/- 0.05) of the cuticular cohort, 55% (frequency +/- SE, 0.55 +/- 0.01) of the more typical AMD cases, and 34% (frequency +/- SE, 0.34 +/- 0.02) of controls. The association between the cuticular drusen phenotype and the histidine allele was highly significant (P = .003; odds ratio, 2.0; 95% confidence interval, 1.21-3.07; vs AMD cases P<.001; odds ratio 4.54; 95% confidence interval, 2.79-7.50; vs controls). Genotype distribution between the 3 groups was similarly significant (P<.001)., Conclusion: The cuticular drusen phenotype is highly associated with the Tyr402His variant of the complement factor H (CFH) gene. The significantly higher histidine allele frequency in this group compared with the typical AMD cohort suggests that the complement cascade may play a greater role in the pathogenesis of the cuticular drusen subtype than in AMD as a whole., Clinical Relevance: The c.1204T>C, p.Tyr402His allelic variant in the CFH gene is associated with a 3-fold increased risk for AMD. A high frequency of the histidine allele has also been noted in patients with membranoproliferative glomerulonephritis type II.

Age-related macular degeneration (AMD) is the most common cause of irreversible visual loss in the developed world. Previous studies have demonstrated that the c.1204T>C, p.Tyr402His allelic variant in the complement factor H (CFH) gene is associated with an approximately three-fold increased risk for AMD in Caucasians of predominantly European descent. Both the prevalence as well as the phenotypic spectrum of AMD varies widely among persons of different ethnicities. We hypothesized that populations with a lower prevalence of AMD might also have a lower prevalence of the CFH risk allele. In this study we sought to determine the frequency of this sequence variant in control populations of Caucasians, African Americans, Hispanics, Somalis, and Japanese. Normal control populations were assembled for each ethnic group: Caucasian (n=148), Somali (n=128), African American (n=75), Hispanic (n=81), and Japanese (n=82). Individuals were genotyped using a restriction digest assay and the frequency of the C allele at nucleotide position 1204 of the CFH gene was determined. A bioinformatic approach was used to identify SNPs in linkage disequilibrium with rs1061170 (c.1204T>C, p.Tyr402His) from the human haplotype map project database (HapMap) in order to validate the findings. We found widely discordant frequencies of the risk allele between some of the different ethnic groups: Japanese 0.07+/-0.02, Hispanics 0.17+/-0.03, African-Americans 0.35+/-0.04, Caucasians 0.34+/-0.03, and Somalis 0.34+/-0.03. Allele frequencies generated by analysis of the HapMap database were consistent with these findings. This study suggests that there are other yet unidentified genetic factors important in the pathogenesis of AMD that may mitigate the effects of c.1204T>C, p.Tyr402His variant., ((c) 2006 Wiley-Liss, Inc.)

Purpose: Choroidal neovascularization (CNV) is a complication of multiple eye diseases, including age-related macular degeneration, that usually results in irreversible vision loss. It is characterized by proliferation and growth of choroidal blood vessels through Bruch's membrane into the subpigment epithelial and/or subretinal space. The purpose of this study was to characterize the carbohydrate groups associated with CNV by lectin histochemistry., Methods: Frozen sections from three human eyes with CNV (two fixed eyes and one unfixed eye) were prepared. Sections containing choroidal neovascular membranes were incubated with a battery of biotinylated lectins directed against a number of distinct oligosaccharide moieties. Lectin labeling of the vessels in CNV was visualized with avidin-Texas red., Results: Several carbohydrate groups were preferentially associated with the vascular elements in CNV. Glycoconjugates that react with lectins derived from wheat germ, soybean, and hairy vetch seed (sWGA, SBA, and VVA, respectively) all showed reactivity with CNV vessels that was higher than the labeling of the surrounding matrix. SBA and sWGA also reacted with CNV vessels at low concentrations at which normal retinal and choroidal vessels were largely unlabeled., Conclusions: Choroidal neovascular membranes possess a distinct set of carbohydrate moieties. These data may be valuable in understanding endothelial cell biology in CNV.

Purpose: To elucidate the phenotypic and biochemical characteristics of a novel mutation associated with autosomal dominant cone-rod dystrophy (adCORD)., Methods: Twenty-three family members of a CORD pedigree underwent clinical examinations, including visual acuity tests, standardized full-field ERG, and fundus photography. Genomic DNA was screened for mutations in GCAP1 exons using DNA sequencing and single-strand conformational polymorphism (SSCP) analysis. Function and stability of recombinant GCAP1-L151F were tested as a function of [Ca(2+)], and its structure was probed by molecular dynamics., Results: Affected family members experienced dyschromatopsia, hemeralopia, and reduced visual acuity by the second to third decade of life. Electrophysiology revealed a nonrecordable photopic response with later attenuation of the scotopic response. Affected family members harbored a C-->T transition in exon 4 of the GCAP1 gene, resulting in an L151F missense mutation affecting the EF hand motif 4 (EF4). This change was absent in 11 unaffected family members and in 100 unrelated normal subjects. GCAP1-L151F stimulation of photoreceptor guanylate cyclase was not completely inhibited at high physiological [Ca(2+)], consistent with a lowered affinity for Ca(2+)-binding to EF4., Conclusions: A novel L151F mutation in the EF4 hand domain of GCAP1 is associated with adCORD. The clinical phenotype is characterized by early cone dysfunction and a progressive loss of rod function. The biochemical phenotype is best described as persistent stimulation of photoreceptor guanylate cyclase, representing a gain of function of mutant GCAP1. Although a conservative substitution, molecular dynamics suggests a significant change in Ca(2+)-binding to EF4 and EF2 and changes in the shape of L151F-GCAP1.

Purpose: To describe the features of atypical pigment dispersion in a child., Design: Observational case report., Methods: Retrospective chart review., Results: An 8-year-old boy presented with midperipheral iris transillumination defects, iris backbowing, heavy pigmentation of the trabecular meshwork, and elevated intraocular pressure. He also had several atypical features including emmetropia, mild posterior subcapsular cataract, small pupils, and peripheral anterior synechiae., Conclusion: Pigment dispersion in childhood is unusual and may have different features from those classically described in adults.

A 37-year-old man presented with fever and a red, painful right eye. He had proptosis, conjunctival chemosis, and ophthalmoplegia OD. The patient had extremely poor dentition and had self decompressed a dental abscess prior to admission. Magnetic resonance imaging of the brain and orbital revealed extraocular muscle engorgement and a dilated superior ophthalmic vein OD. Orbital echography revealed a lack of flow in the right superior ophthalmic vein. An extensive hematologic evaluation for infection and inflammation was negative. A chest radiograph showed a lung abscess for which he received intravenous antibiotics. Over time, the periorbital erythema, ophthalmoplegia, proptosis, and pain resolved. Repeat MRI showed resolution of the orbital findings and repeat chest x-ray showed resolution of the left upper lobe abscess.

Purpose: To describe diplopia as the initial manifestation of the Burkitt lymphoma., Design: Observational case report., Methods: Retrospective chart review., Results: A 53-year-old human immunodeficiency virus (HIV)-positive man presented with a severe headache associated with binocular diplopia. Flow cytometric analysis of the cerebrospinal fluid demonstrated a monoclonal B-lymphoid cell population consistent with the Burkitt lymphoma., Conclusion: Ophthalmologists should be aware that diplopia may be the presenting manifestation of Burkitt lymphoma in an immunocompromised patient.

Background: Abdominal aortic aneurysms (AAA) are characterized by both increases in proteolysis and changes in the biosynthesis of the extracellular matrix (ECM) proteins. Proteoglycans are important components of the ECM, particularly the small proteoglycans, biglycan and decorin. Biglycan and decorin regulate cell proliferation and collagen assembly. Therefore, the purpose of this study is to quantify the levels of mRNA for biglycan and decorin in normal aorta (Na) and AAA., Materials and Methods: Northern blot hybridization and competitive polymerase chain reaction using gene-specific external standards were used to quantify mRNA levels of bigylcan and decorin in RNA derived from AAA and NA. Results are expressed as a percentage of glyceraldehyde-3-phosphate dehydrogenase or normalized to ribosomal RNA content and compared using the unpaired t test., Results: A statistically significant 15-fold decrease in biglycan mRNA expression was observed in AAA compared to NA (176.9% vs 11.8%, P < 0.001). In contrast to biglycan, the decorin mRNA expression is unchanged in AAA compared to NA., Conclusions: The marked decrease in biglycan mRNA levels is unique to aneurysmal disease of the aorta. In atherosclerosis and restenosis, biglycan expression is increased in comparison with normal artery. This decrease in biglycan expression may reflect important regulatory changes specific for the AAA. Furthermore, a decrease in biglycan gene expression and biosynthesis could have a broad impact on the physiology and matrix architecture of the aorta.

A modification of Lerke enzymic method for histamine determination has been realized. We have observed a modification of the optimum wavelength depending on incubation time at constant temperature of 37°C. A wavelength of 580 nm is recommended, with a incubation time of 15 min. At these conditions the linearity was observed among 1 and 25 ppm (μg/g) of histamine. This technique is a reliable method of screening in food samples and bacterial strains, with low sample preparations requirements, a short incubation time and a low cost.