Sensitivity and specificity of frequency-doubling technology, tendency-oriented perimetry, SITA Standard and SITA Fast perimetry in perimetrically inexperienced individuals

Purpose: To evaluate the sensitivity and specificity of the screening modes of frequency-doubling technology (FDT), tendency-oriented perimetry (TOP), SITA Standard (SS) and SITA Fast (SF) in perimetrically inexperienced individuals. Methods: One eye of 64 glaucoma patients and 53 normal subjects who had never undergone automated perimetry were tested with programs C-20-5 (FDT), G1 (TOP) and 24-2 (SS and SF). The gold standard for glaucoma was the presence of a typical glaucomatous optic disc appearance on stereoscopic examination (judged by a glaucoma expert), and intraocular pressure (IOP) > 21 mmHg. The test order among strategies was randomized for each subject. To define an abnormal visual field, we applied three criteria for SS and SF and two criteria for TOP and FDT, all of which have been previously described in the literature. Sensitivities and specificities among the different criteria were compared using the Cochran test. Results: Frequency-doubling technology showed the shortest mean test duration, followed by TOP, SF and SS (p < 0.05). Sensitivity ranges were 87.5-89.1% for SS, 92.2-93.8% for SF, 87.5-89.1% for TOP, and 82.8-85.9% for FDT (p = 0.34). Specificity ranges were 73.6-83% for FDT, 56.6-62.3% for TOP, 60.4-69.8% for SF and 66.0-71.7% for SS. The specificity obtained with criterion 2 for FDT (based on the presence of two or more abnormal locations regardless of the severity of abnormal points) was higher than those measured with the other strategies (p < 0.01). Conclusion: When testing individuals with no perimetric experience, moderate sensitivities and specificities should be expected, regardless of the strategy chosen.84334535

Evaluation of the Efficacy and Safety of Botulinum Toxin Type A to Induce Temporary Ptosis in Dogs

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Purpose: To verify the safety and efficacy of botulinum toxin type A (BoNT/A) to promote protective ptosis in dogs. Methods: In this prospective interventional study, a total of 10 dogs underwent transcutaneous anterior chemodenervation of levator palpebral superioris with 15U of BoNT/A. The systemic changes, ocular mobility, visual function, intraocular pressure (IOP), tear production, and the onset, degree, and duration of ptosis were evaluated on a daily basis during the first 7 days and on days 14, 21, and 28 after application. Results: The onset of the clinical effect was observed between 2 and 3 days after application of the toxin; the time taken for maximum ptosis to develop varied from 4 to 7 days (mean 5 days) and the average duration of the toxin effect was 21 days. The mean percentage reduction in palpebral fissure height was 42.859% (SD +/- 35.714%-59.821%). There was not a statistically significant difference in IOP before and after the BoNT/A application (P = 0.974), or lacrimal production evaluation (P = 0.276). There was no change in ocular mobility and no other adverse effect was observed in association with the administration of the study drug. Conclusion: The application of BoNT/A into the levator palpebral superioris muscle in dogs was effective and safe to promote protective ptosis with a temporary covering of the cornea.294431436Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Endophthalmitis caused by Agrobacterium radiobacter

Infections due to Agrobacterium radiobacter are rare. This study reports 2 cases of A. radiobacter endophthalmitis. To the authors' knowledge, these are only the second and third reported cases of endophthalmitis caused by this Gram-negative rod.354182641041

A novel mutation in the GJA1 gene in a family with oculodentodigital dysplasia

Objectives: To describe a Brazilian family with oculodentodigital dysplasia (ODDD) and to screen for mutations in the gap junction protein alpha I (GJA1) gene in this family. Methods: Twelve members of a 3-generation family with ODDD underwent screening for mutations of the GJA1 gene and a comprehensive ophthalmic examination. We defined ODDD on the basis of clinical characteristics described in this syndrome (microdontia, caries, enamel hypoplasia, thin nose, and syndactyly) and eye abnormalities such as microphthalmos, iris atrophy, and glaucoma. Direct sequencing of the GJA1 gene was performed using DNA collected from peripheral blood. A control group of 60 healthy individuals underwent evaluation by means of enzyme digestion. Results: Among the 8 members of this family who were characterized as having ODDD, 2 showed chronic angle-closure glaucoma, and 1 had open-angle glaucoma. A new mutation in the GJA1 gene was identified, consisting of a change from proline to histidine at codon 59. This mutation segregated through members with the ODDD phenotype. Analysis of the control group by means of restriction fragment length polymorphism (MvaI enzyme) did not disclose this mutation. Conclusion: Our results demonstrate a new mutation (P59H) in the GJ1A gene, identified in a family with ODDD syndrome. Clinical Relevance: The presence of different forms of glaucoma in families with ODDD may indicate a new mutation in the GJA1 gene.123101422142

Mutation analysis of CRYAA, CRYGC, and CRYGD associated with autosomal dominant congenital cataract in Brazilian families

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Purpose: Congenital cataracts are one of the most treatable causes of visual impairment and blindness during infancy. Approximately 50% of all congenital cataract cases may have a genetic cause. Once there is an intimate relationship between crystallin genes and lens transparency, they are excellent candidate genes for inherited cataract. The purpose of this study was to investigate mutations in alpha A-crystallin (CRYAA), gamma C-crystallin (CRYGC), and gamma D-crystallin (CRYGD) in Brazilian families with nuclear and lamellar autosomal dominant congenital cataract. Methods: Eleven Brazilian families were referred to the Santa Casa de Sao Paulo Ophthalmology Department. The coding regions and intron/exon boundaries of CRYAA, CRYGC, and CRYGD were amplified by polymerase chain reaction (PCR) and directly sequenced. Mutation screening was performed in the control group by restriction digestion. Results: Two mutations were observed in different families (Family 4 and Family 10), one is a new mutation (Y56X) in CRYGD and the other a previously reported mutation (R12C) in CRYAA that is correlated with a different phenotype. Genetic analysis revealed previously described polymorphisms in CRYAA (D2D) and CRYGD (Y17Y and R95R). A new polymorphism in CRYGC (S119S) was identified only in Family 1. The mutations as well as the new polymorphism were not observed in the control group. Conclusions: In conclusion, we report a novel nonsense mutation (Y56X) in CRYGD and a previously reported missense mutation (R12C) in CRYAA associated with nuclear cataract in Brazilian families. Both tyrosine in amino acid 56 in CRYGD and arginine in amino acid 12 in CRYAA have been highly conserved throughout evolution in different species. A new polymorphism (S119S) in CRYGC was also observed in one family. The analysis of nine families excluded possible mutations in the crystallin genes, suggesting that other genes could be involved with congenital cataract.1579-81793800Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)FAP-SCSP (Santa Casa de Sao Paulo School of Medicine, Sao Paulo, Brazil)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Structural assessment of PITX2, FOXC1, CYP1B1, and GJA1 genes in patients with Axenfeld-Rieger syndrome with developmental glaucoma

PURPOSE. Axenfeld-Rieger (AR) is an autosomal dominant disorder with phenotypic heterogeneity characterized by anterior segment dysgenesis, facial bone defects, and redundant periumbilical skin. The PITX2 gene, on chromosome 4q25, and the FOXC1 gene, on chromosome 6p25, have been implicated in the different phenotypes of the syndrome through mutational events. Recently, the CYP1B1 gene was found to be associated with Peters' anomaly, and the gene associated with oculoden-todigital dysplasia syndrome, which presents some similarities with AR, was identified ( connexin 43 - GJA1 gene). The purpose of this study was to evaluate PITX2, FOXC1, CYP1B1, and GJA1 gene mutations in Brazilian families with AR. METHODS. Eight unrelated patients affected by AR ( all eight with glaucoma and three with systemic manifestations) and their families were ophthalmologically evaluated and their blood was collected for DNA extraction purposes. The coding regions of PITX2, FOXC1, CYP1B1, and GJA1 genes were completely evaluated through direct sequencing. RESULTS. The frequency of mutations in the FOXC1, GJA1, PITX2, and CYP1B1 genes in this study were 25%, 12.5%, 0% and 0%, respectively. In the FOXC1 gene, two GGC triplet insertions (GGC375ins and GGC447ins) defined as a polymorphism, and two new mutations-a deletion ( 718 to 719delCT) and a nonsense mutation (Trp152STOP)-were identified. One polymorphism (Ala253Val) was identified in the GJA1 gene in the same family presenting the Trp152STOP mutation in the FOXC1 gene. In this family harboring both structural alterations, two patients who carried the GJA1 ( Ala253Val) and FOXC1 ( Trp152STOP) mutations developed less severe glaucoma compared with family members presenting the FOXC1 (Trp152STOP) mutation alone. CONCLUSIONS. Two new structural alterations in the FOXC1 gene and a polymorphism in the GJA1 gene were first described in Brazilian patients with AR and developmental glaucoma. A polymorphism in the GJA1 gene (Ala253Val), for the first time identified in association with AR, raises the possibility of its participation as a modifier gene.O TEXTO COMPLETO DESTE ARTIGO, ESTARÁ DISPONÍVEL À PARTIR DE FEVEREIRO DE 2015.4751803180

Molecular genetics of primary congenital glaucoma in Brazil

PURPOSE. To determine the distribution of CYP1B1 gene mutations in Brazilian patients with primary congenital glaucoma (PCG). METHODS. PCG diagnosis was established by presence of buphthalmos in at least one affected eye and associated high intraocular pressures before the age of 3 years. CYP1B1 mutation screening of 52 patients with PCG was performed by SSCP and direct sequencing of PCR fragments. RESULTS. Eleven mutations, four of which are novel, were observed in 26 (50%) individuals. A new frameshift mutation (4340delG) was observed in 20.2% of all individuals screened. These individuals had early-onset, bilateral glaucoma that necessitated multiple surgical interventions. CYP1B1 mutations were twice as frequent in affected individuals of European descent as in individuals of African descent. Analysis of six intragenic single nucleotide polymorphisms (SNPs) established 5'-C-C-G-G-T-A-3' as the most common haplotype among the affected Brazilian individuals. A nonsense mutation (W57X) previously reported in an individual with Peters anomaly (compound heterozygote) was also observed in two individuals with PCG but combined with different Mutations. A newly developed SSCP assay enabled us to detect all DNA mutations and polymorphisms previously detected by direct sequencing. CONCLUSIONS. Our results indicate that CYP1B1 mutations may be responsible for half of cases of PCG in the Brazilian population. The SNP haplotype 5'-C-C-G-G-T-A-3' was associated with the majority of CYP1B1 mutations. This haplotype harbors the high-activity V432 allele, which is emerging as a putative susceptibility factor in several cancers.O TEXTO COMPLETO DESTE ARTIGO, ESTARÁ DISPONÍVEL À PARTIR DE FEVEREIRO DE 2015.4361820182

Novel human CRYGD rare variant in a Brazilian family with congenital cataract

Purpose: To describe a novel polymorphism in the gamma D-crystallin (CRYGD) gene in a Brazilian family with congenital cataract. Methods: A Brazilian four-generation family was analyzed. The proband had bilateral lamellar cataract and the phenotypes were classified by slit lamp examination. Genomic DNA was extracted from peripheral blood and coding regions and intron/exon boundaries of the alpha A-crystallin (CRYAA), gamma C-crystallin (CRYGC), and CRYGD genes were amplified by polymerase chain reaction and directly sequenced. Results: Sequencing of the coding regions of CRYGD showed the presence of a heterozygous A -> G transversion at c. 401 position, which results in the substitution of a tyrosine to a cysteine (Y134C). The polymorphism was identified in three individuals, two affected and one unaffected. Conclusions: A novel rare variant in CRYGD (Y134C) was detected in a Brazilian family with congenital cataract. Because there is no segregation between the substitution and the phenotypes in this family, other genetic alterations are likely to be present.17238-392207221

New mutation in the myocilin gene segregates with juvenile-onset open-angle glaucoma in a Brazilian family

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Mutations in the myocilin gene (MYOC) account for most cases of autosomal dominant juvenile-onset open-angle glaucoma (JOAG), an earlier and more severe form of POAG. We accessed seven members of a Brazilian JOAG family by clinical and molecular investigation. Four out of seven family members were diagnosed with JOAG. All of these patients presented high intraocular pressure and two of them were bilaterally blind. The disease onset varied from 20 to 30 years old. There was a nine-year-old family member who had not yet manifested the disease, although he was also a carrier of the mutation. Ophthalmologic examination included: evaluation of the visual field and optic disc, intraocular pressure measurement, and gonioscopy. The three exons and intron/exon junctions of the MYOC gene were screened for mutations through direct sequencing of PCR-amplified DNA fragments. Mutation screening revealed an in-frame mutation in the third exon of the MYOC gene: an insertion of six nucleotides between the cDNA positions 1187 and 1188 (c.1187_1188insCCCAGA, p.D395_E396insDP). This mutation presented an autosomal dominant pattern of inheritance, segregating with the disease in four family members for three generations, and it was absent in 60 normal controls. We also performed a computational structure modeling of olfactomedin-like domain of myocilin protein and conducted in silico analysis to predict the structural changes in the myocilin protein due to the presence of the mutation. These findings may be important for future diagnosis of other presymptomatic family members, as well as for the increase of the panel of MYOC mutations and their effects on phenotype. (C) 2013 Elsevier B.V. All rights reserved.52315057Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAPESP [2009/15223-0

CYP1B1 Gene Analysis in Primary Congenital Glaucoma Brazilian Patients Novel Mutations and Association With Poor Prognosis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Purpose: To determine the spectrum of CYP1B1 gene mutations in Brazilian patients with primary congenital glaucoma, and to correlate the presence of alterations in the CYP1B1 gene sequence with clinical aspects of the disease. Materials and Methods: Thirty nonrelated patients with primary congenital glaucoma were studied. Molecular analysis consisted of the codifying region sequencing (exons 2 and 3) and intron/exon boundaries. Results: CYP1B1 gene mutations were present in 9 (30%) of the 30 patients. The structural changes in the CYP1B1 gene previously described in the literature and observed in our study were Q19X, P437L, A443G, g.4340delG, g.7901_79013delGAGTGCAGGCAGA, g.8182delG, and g.8214_8215delG. Three new mutations were observed: 4635delT, 4523delC, and L378Q, in addition to 3793T --> C, R48G, A119S, L432V, D449D, and N453S polymorphisms. Patients carrying CYP1B1 gene mutations needed more surgical procedures to control intraocular pressure, either when both eyes were evaluated (P = 0.003) or when the worst eye of the patient was analyzed (P = 0.011). In relation to the number of affected eyes, all patients with mutations (n = 9/9) developed bilateral glaucoma, whereas 11/21 patients without mutations in the CYP1B1 gene had bilateral glaucoma (P = 0.013). Conclusions: In this group of primary congenital glaucoma patients, a 30% mutation frequency in the CYP1B1 gene was observed. The presence of mutations was associated with a more severe form of the disease, requiring more surgeries for intraocular pressure control and with a higher rate of bilateral cases.193176182FAP Santa Casa de Sao PauloFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)FAPESP [02/11575-0