The effect of copy number variations in chromosome 16p on body weight in patients with intellectual disability.

Syndromic monogenic obesity is a rare and severe early-onset form of obesity. It is characterized by intellectual disability, congenital malformations, and/or dysmorphic facies. The diagnosis of patients is challenging due to the genetic heterogenicity of this condition. However, the use of microarray technology in combination with public databases has been successful on genotype-phenotype correlations, especially for body mass index (BMI) alteration. In this study, the relationship between copy number variations (CNVs) detected by microarray mapping on 16p region and BMI alterations in syndromic patients were assessed. In order to achieve this goal, 680 unrelated Spanish children with intellectual disability were included. 16p region was characterized by using microarray platforms. All detected variants were classified as: (I) one previously non-described 10-Mb duplication in 16p13.2p12.3 region considered causal of intellectual disability and severe overweight, and (II) eleven 16p11.2 CNVs of low prevalence but with recurrence in syndromic patients with severe BMI alteration (nine proximal and two distal). Proximal 16p11.2 CNVs have a dose-dependent effect: underweight in carriers of duplication and obesity in carriers of deletion. KCTD13 was identified as a possible candidate gene for BMI alteration on proximal syndromes, whereas SH2B1 gene was identified as candidate for distal syndromes. The results shown in this paper suggest that syndromic patients could constitute a reliable model to evaluate hypothalamic satiety and obesity disorders as well as generate a wide expectation for primary prevention of comorbidities. Furthermore, 16p13.2p12.3 showed to be an important region on the regulation of body fatness.

Neurodevelopmental Disorders and Array-Based Comparative Genomic Hybridization: Sensitivity and Specificity using a Criteria Checklist for Genetic Test Performance.

BACKGROUND: Array-based comparative genomic hybridization (aCGH) is a molecular analysis method for identifying chromosomal anomalies or copy number variants (CNVs) correlating with clinical phenotypes. The aim of our study was to identify the most significant clinical variables associated with a positive outcome of aCGH analyses to develop a simple predictive clinical score. METHODS: We conducted a cross-sectional study in a tertiary center comparing the genotype and phenotype of the cases. A score was developed using multivariate logistic regression. The best score cutoff point, sensitivity, specificity, positive and negative predictive values, and area under the curve were calculated with the receiver operating characteristic curve. RESULTS: aCGH identified structural chromosomal alterations responsible for the disorder in 13.7% (95% confidence interval [CI]: 10.9-16.5) of our sample (570 patients analyzed by aCGH). Based on the most frequent phenotypic characteristics among patients with a pathogenic CNV, we have created a checklist with the following items: alteration of the cranial perimeter, stature < percentile (p) 3, weight < p3, presence of brain malformations, ophthalmological malformations, two or more dysmorphic features in the same patient, and autism spectrum disorder diagnosis. Using a score ≥1.5 as the cutoff point for the test, we obtained a sensitivity of 82.4% (95% CI: 73.1-91.8) and a specificity of 54.2% (95% CI: 49.7-58.7). CONCLUSION: All individuals with a score of 1.5 or higher should be genetically screened by aCGH. This approach can improve clinical indications for aCGH in patients with neurodevelopmental disorders, but the scoring system should be validated in an external group.

Genotype-phenotype correlations in hypertrophic cardiomyopathy: a multicenter study in Portugal and Spain of the TPM1 p.Arg21Leu variant.

INTRODUCTION AND OBJECTIVES: TPM1 is one of the main hypertrophic cardiomyopathy (HCM) genes. Clinical information on carriers is relatively scarce, limiting the interpretation of genetic findings in individual patients. Our aim was to establish genotype-phenotype correlations of the TPM1 p.Arg21Leu variant in a serie of pedigrees. METHODS: TPM1 was evaluated by next-generation sequencing in 10 561 unrelated probands with inherited heart diseases. Familial genetic screening was performed by the Sanger method. We analyzed TPM1 p.Arg21Leu pedigrees for cosegregation, clinical characteristics, and outcomes. We also estimated the geographical distribution of the carrier families in Portugal and Spain. RESULTS: The TPM1 p.Arg21Leu variant was identified in 25/4099 (0.61%) HCM-cases, and was absent in 6462 control individuals with other inherited cardiac phenotypes (P<.0001). In total, 83 carriers (31 probands) were identified. The combined LOD score for familial cosegregation was 3.95. The cumulative probability of diagnosis in carriers was 50% at the age of 50 years for males, and was 25% in female carriers. At the age of 70 years, 17% of males and 46% of female carriers were unaffected. Mean maximal left ventricular wall thickness was 21.4 ±7.65mm. Calculated HCM sudden death risk was low in 34 carriers (77.5%), intermediated in 8 (18%), and high in only 2 (4.5%). Survival free of cardiovascular death or heart transplant was 87.5% at 50 years. Six percent of carriers were homozygous and 18% had an additional variant. Family origin was concentrated in Galicia, Extremadura, and northern Portugal, suggesting a founder effect. CONCLUSIONS: TPM1 p.Arg21Leu is a pathogenic HCM variant associated with late-onset/incomplete penetrance and a generally favorable prognosis.

ABCB4 variants in adult patients with cholestatic disease are frequent and underdiagnosed.

BACKGROUND: Heterozygous ABCB4 variants are not routinely tested in adults with cholestasis because of their supposed rarity and high costs. METHODS: Nineteen adult patients presenting with unexplained cholestasis, and/or recurrent gallstones were included; genotyping was not done in five due to lack of health insurance approval. RESULTS: heterozygous ABCB4 variants were identified in seven patients, followed by cascade testing of 12 family members: one patient underwent liver transplantation at age 40 for end-stage liver disease; one had compensated cirrhosis; all symptomatic adults had gallstones, including four with low phospholipid-associated cholelithiasis; four had intrahepatic cholestasis of pregnancy; all children and one 54-year old female were asymptomatic. Genotype: Families A and C: c.2211G>A (p.Ala737=) combined with c.959C>T (p.Ser320Phe) in one subject; Family B: c.1130T>C (p.Ile377Thr); Family D: large deletion removing ABCB4 exons 1-4 plus ABCB1, RUNDC3B, SLC25A40, DBF4, ADAM22 exons 1-3; Family E: c.1565T>C (p.Phe522Ser) ; Family F: c.1356+2T>C combined with c.217C>G (p.Leu73Val). All patients responded to ursodeoxycholic acid. CONCLUSIONS: We found ABCB4 variants in half of the adults with unexplained cholestasis and/or recurrent gallstones presenting at our center, suggesting that this condition is underdiagnosed and undertreated, with serious consequences not only for the patients and their families, but also in terms of healthcare costs.

Clear cell clusters in the kidney: a rare finding that should not be misdiagnosed as renal cell carcinoma.

Clear cytoplasm is a major characteristic feature of most malignant renal neoplasms. Benign clear cells in the renal parenchyma, usually histiocytes, can occasionally be found, but they are infrequently of an epithelial nature. We report histological, immunohistochemical, ultrastructural, and cytogenomic features of clear epithelial cell clusters incidentally found in four kidney specimens. Multiple microscopic clear cell clusters were present in the cortex, often in subcapsular location. They were composed of large epithelial cells with strikingly clear cytoplasm, without nuclear atypia, arranged in solid nests, and some tubules with narrow lumina. Immunohistochemically, they were positive for AE1AE3, PAX 8, EMA, kidney-specific cadherin, cytokeratin 7, E cadherin, and CD117, with focal immunoreactivity for CD10. Carbonic anhydrase IX, vimentin, and markers related to apoptosis and proliferation were negative. Ultrastructurally, the cytoplasms were enlarged and poor in organelles, showing ballooning degeneration. Array comparative genomic hybridization showed no chromosomal gains or losses. Clear cell clusters constitute a rare finding in the kidney and must be differentiated from benign lesions (ectopic adrenal tissue, osmotic tubulopathy, histiocytic clusters, renal adenomas) and renal cell carcinomas. Clear cell clusters appear to be generated from "endocrine-type" atrophic tubules whose cells are enlarged due to intracellular oedema. Immunohistochemistry shows a distal nephron phenotype with a limited expression of a proximal marker, CD10. Coexisting chronic renal disease or ischemic conditions seem to be related to the development of clear cell clusters. Pathological, ultrastructural, and cytogenomic features do not support a preneoplastic nature of this lesion, at least in the cases studied here.

Comprehensive genomic diagnosis of non-syndromic and syndromic hereditary hearing loss in Spanish patients.

BACKGROUND: Sensorineural hearing loss (SNHL) is the most common sensory impairment. Comprehensive next-generation sequencing (NGS) has become the standard for the etiological diagnosis of early-onset SNHL. However, accurate selection of target genomic regions (gene panel/exome/genome), analytical performance and variant interpretation remain relevant difficulties for its clinical implementation. METHODS: We developed a novel NGS panel with 199 genes associated with non-syndromic and/or syndromic SNHL. We evaluated the analytical sensitivity and specificity of the panel on 1624 known single nucleotide variants (SNVs) and indels on a mixture of genomic DNA from 10 previously characterized lymphoblastoid cell lines, and analyzed 50 Spanish patients with presumed hereditary SNHL not caused by GJB2/GJB6, OTOF nor MT-RNR1 mutations. RESULTS: The analytical sensitivity of the test to detect SNVs and indels on the DNA mixture from the cell lines was > 99.5%, with a specificity > 99.9%. The diagnostic yield on the SNHL patients was 42% (21/50): 47.6% (10/21) with autosomal recessive inheritance pattern (BSND, CDH23, MYO15A, STRC [n = 2], USH2A [n = 3], RDX, SLC26A4); 38.1% (8/21) autosomal dominant (ACTG1 [n = 3; 2 de novo], CHD7, GATA3 [de novo], MITF, P2RX2, SOX10), and 14.3% (3/21) X-linked (COL4A5 [de novo], POU3F4, PRPS1). 46.9% of causative variants (15/32) were not in the databases. 28.6% of genetically diagnosed cases (6/21) had previously undetected syndromes (Barakat, Usher type 2A [n = 3] and Waardenburg [n = 2]). 19% of genetic diagnoses (4/21) were attributable to large deletions/duplications (STRC deletion [n = 2]; partial CDH23 duplication; RDX exon 2 deletion). CONCLUSIONS: In the era of precision medicine, obtaining an etiologic diagnosis of SNHL is imperative. Here, we contribute to show that, with the right methodology, NGS can be transferred to the clinical practice, boosting the yield of SNHL genetic diagnosis to 50-60% (including GJB2/GJB6 alterations), improving diagnostic/prognostic accuracy, refining genetic and reproductive counseling and revealing clinically relevant undiagnosed syndromes.

Formin Homology 2 Domain Containing 3 (FHOD3) Is a Genetic Basis for Hypertrophic Cardiomyopathy.

BACKGROUND: The genetic cause of hypertrophic cardiomyopathy remains unexplained in a substantial proportion of cases. Formin homology 2 domain containing 3 (FHOD3) may have a role in the pathogenesis of cardiac hypertrophy but has not been implicated in hypertrophic cardiomyopathy. OBJECTIVES: This study sought to investigate the relation between FHOD3 mutations and the development of hypertrophic cardiomyopathy. METHODS: FHOD3 was sequenced by massive parallel sequencing in 3,189 hypertrophic cardiomyopathy unrelated probands and 2,777 patients with no evidence of cardiomyopathy (disease control subjects). The authors evaluated protein-altering candidate variants in FHOD3 for cosegregation, clinical characteristics, and outcomes. RESULTS: The authors identified 94 candidate variants in 132 probands. The variants' frequencies were significantly higher in patients with hypertrophic cardiomyopathy (74 of 3,189 [2.32%]) than in disease control subjects (18 of 2,777 [0.65%]; p < 0.001) or in the gnomAD database (1,049 of 138,606 [0.76%]; p < 0.001). FHOD3 mutations cosegregated with hypertrophic cardiomyopathy in 17 families, with a combined logarithm of the odds score of 7.92, indicative of very strong segregation. One-half of the disease-causing variants were clustered in a small conserved coiled-coil domain (amino acids 622 to 655); odds ratio for hypertrophic cardiomyopathy was 21.8 versus disease control subjects (95% confidence interval: 1.3 to 37.9; p < 0.001) and 14.1 against gnomAD (95% confidence interval: 6.9 to 28.7; p < 0.001). Hypertrophic cardiomyopathy patients carrying (likely) pathogenic mutations in FHOD3 (n = 70) were diagnosed after age 30 years (mean 46.1 ± 18.7 years), and two-thirds (66%) were males. Of the patients, 82% had asymmetric septal hypertrophy (mean 18.8 ± 5 mm); left ventricular ejection fraction <50% was present in 14% and hypertrabeculation in 16%. Events were rare before age 30 years, with an annual cardiovascular death incidence of 1% during follow-up. CONCLUSIONS: FHOD3 is a novel disease gene in hypertrophic cardiomyopathy, accounting for approximately 1% to 2% of cases. The phenotype and the rate of cardiovascular events are similar to those reported in unselected cohorts. The FHOD3 gene should be routinely included in hypertrophic cardiomyopathy genetic testing panels.

Tool use by Amazonian capuchin monkeys during predation on caiman nests in a high-productivity forest.

Descriptions of new tool-use events are important for understanding how ecological context may drive the evolution of tool use among primate traditions. Here, we report a possible case of the first record of tool use by wild Amazonian capuchin monkeys (Sapajus macrocephalus). The record was made by a camera trap, while we were monitoring caiman nest predation at Mamirauá Reserve in Central Amazonia. An adult individual was registered in a bipedal posture, apparently using a branch as a shovel to dig eggs out of a nest. Caiman eggs are frequently depredated by opportunistic animals, such as the capuchin monkeys. As the Mamirauá Reserve is covered by a high-productivity forest, and caiman eggs are a high-quality food resource seasonally available on the ground, we believe that tool use by capuchins is more likely to be opportunity driven, rather than necessity driven, in our study site.

[Study of mycobiota in the healthy conjunctiva of diabetics who reside in the urban area of the city of São Paulo, Brazil]. / Estudo da micobiota em conjuntiva sadia de diabéticos, residentes na área urbana da cidade de São Paulo-Brasil.

PURPOSE: To determine the mycobiota of the healthy conjunctiva in diabetic individuals, according to diabetes type, age, sex, disease time, type of treatment, and stage of diabetic retinopathy of the individuals. To identify the anemophilus mycobiota in the sampling rooms. METHODS: A cross-sectional study was carried out on 803 diabetics who reside in the urban area of São Paulo-SP/Brazil. Sabouraud's dextrose agar culture with chloramphenicol was used for primoisolation, and the key of De Hoog was used to identify filamentous fungi. RESULTS: Of the evaluated diabetics, 6.6% (53/803) presented type 1 diabetes and 93.4% (750/803) type 2. The positive cultures for fungi in the conjunctiva of diabetics was 4.2% (34/803), with 1.9% (1/53) in type 1 diabetics and 4.4% (33/740) in type 2 diabetics (p=0.720). With respect to the presence or not of isolated fungi, there was no statistically significant association regarding age (p=0.575), sex (p=0.517), disease time (p=0.633), type of treatment (p=0.422), and diabetic retinopathy stage (p=0.655) of the tested individuals. The identified fungi were all filamentous: Aspergillus spp. represented 59.5% (25/42) of isolations and 47.6% (20/42) of isolated species were Aspergillus niger. Growth of anemophilus fungi occurred in the air of the room and coincidences were observed between the isolated species from the air and those from the conjunctiva. CONCLUSIONS: Presence of mycobiota in healthy conjunctivas of diabetics was identified, with no significant association between the greater number of positive fungi isolations and the type of diabetes, age, sex, disease type, type of treatment, and stage of diabetic retinopathy. In the collection rooms, anemophilus mycobiota was identified.

[Laboratory study of the mycotic infectious eye diseases and factors associated with keratitis]. / Estudo laboratorial das micoses oculares e fatores associados às ceratites.

PURPOSE: To analyze the frequency and the etiology of mycotic infectious eye diseases diagnosed by culture at the Ocular Diseases Laboratory of the Federal University of São Paulo (LOFT-UNIFESP), São Paulo, SP, Brazil, and the factors associated with fungal keratitis. METHODS: We retrospectively analyzed the 14,391 laboratory requests regarding infectious eye diseases between 1975 and May 2003. For the fungal culture proven examinations we observed the morphologic type of the isolated fungus, the gender and age of the patient, the site of ocular involvement, the year of identification, and the factors associated with keratitis cases, according to the identified etiologic agent. Results were considered significant if p<0.05. RESULTS: We verified 296 (2.05%) ocular mycosis cases with 265 keratitis (89.5%), 27 intraocular infections (9.1%), and 4 infections at other sites (1.3%). Filamentous fungi were isolated in 233 cases (78.7%) 74.7% being male (n=174) and 25.3% female (n=59). Yeasts were isolated in 63 cases (21.3%) 49.2% being male (n=31) and 50.8% female (n=32). Ages varied between 2 and 99 years in the filamentous fungi group and between 14 and 88 years in the yeast group. The most frequently found etiologic agents of the filamentous fungi isolations were Fusarium (n=137; 58.8%), Aspergillus (n=28; 12.0%), and Penicillium (n=12; 5.2%); and in yeasts, Candida (n=59; 93.7%). Regarding keratitis, 95.1% had at least one associated factor. Ocular trauma was significantly associated with filamentous fungi keratitis (p=0.0002); and previous surgeries (p=0.0002), ocular (p=0.0002) and systemic diseases (p=0.0002) with yeast keratitis. The use of topic antibiotics showed association with filamentous fungi keratitis, at a lower statistical significance (p=0.0224). CONCLUSIONS: Laboratory analysis showed predominance of filamentous fungi over the yeasts, and keratitis was the main observed ocular infection. Fungal keratitis involved at least one associated factor in most cases.

Estudo da micobiota em conjuntiva sadia de diabéticos, residentes na área urbana da cidade de São Paulo - Brasil / Study of mycobiota in the healthy conjunctiva of diabetics who reside in the urban area of the city of São Paulo, Brazil

OBJETIVOS: Determinar a micobiota de conjuntiva sadia em indivíduos diabéticos, segundo tipo de diabetes, idade, sexo, tempo de doenca, tipo de tratamento e estádio da retinopatia. Estabelecer a micobiota anemófila nas salas de colheita. MÉTODOS: Estudo transversal de 803 diabéticos residentes na zona urbana de São Paulo - SP/Brasil. Foi usado para primo-isolamento o meio de cultivo ágar Sabouraud dextrose com cloranfenicol e para identificacão dos fungos filamentosos a chave de De Hoog. RESULTADOS: Dos diabéticos avaliados, 6,6 por cento (53/803) apresentavam diabetes tipo 1 e 93,4 por cento (750/803) tipo 2. Os cultivos positivos para fungos em conjuntiva de diabéticos foi 4,2 por cento (34/803), sendo 1,9 por cento (1/53) nos diabéticos tipo 1 e 4,4 por cento (33/740) nos diabetes tipo 2 (p=0,720). Não foi verificada associacão estatisticamente significante quanto à presenca ou não de isolamentos de fungos em relacão idade (p=0,575), sexo (p=0,517), tempo de doenca (p=0,633), tipo de tratamento (p=0,422) e estádio de retinopatia diabética (p=0,655) desses indivíduos. Todos os fungos identificados foram filamentosos: Aspergillus spp. representou 59,5 por cento (25/42) dos isolamentos sendo 47,6 por cento (20/42) Aspergillus niger. Ocorreu crescimento de fungos anemófilos do ar ambiente da sala, observando-se coincidências entre as espécies isoladas no ar e na conjuntiva. CCONCLUSÕES: Foi identificada presenca de micobiota em conjuntiva sadia de diabéticos, não havendo associacão entre a maior positividade de isolamentos fúngicos e o tipo de diabetes, idade, sexo, tempo de doenca, tipo de tratamento e estádio da retinopatia diabética. Nas salas de colheita foi identificada micobiota anemófila.

Estudo laboratorial das micoses oculares e fatores associados às ceratites / Laboratory study of the mycotic infectious eye diseases and factors associated with keratitis

OBJETIVO: Analisar a freqüência e a etiologia das doenças oculares micóticas diagnosticadas por cultura no Laboratório de Doenças Externas Oculares da Universidade Federal de São Paulo (LOFT-UNIFESP), São Paulo, SP, Brasil e os fatores associados às ceratites fúngicas. MÉTODOS: Estudo retrospectivo de 14.391 solicitações de exames laboratoriais de doenças infecciosas oculares no período compreendido entre 1975 e maio de 2003. Nas comprovações por cultura de micoses oculares foram observados o tipo morfológico do fungo isolado, o sexo e a idade dos pacientes, o local de acometimento da infecção ocular, os anos de identificação e os fatores associados nos casos de ceratite fúngica, conforme o agente etiológico identificado. Considerou-se significante p<0,05. RESULTADOS: Foram verificadas 296 (2,05 por cento) comprovações por cultura de infecção ocular micótica, sendo 265 (89,5 por cento) identificadas como agente etiológico de ceratites, 27 de infecções intra-oculares (9,1 por cento) e 4 de outros locais (1,3 por cento). Fungos filamentosos foram identificados em 233 casos (78,7 por cento), sendo 74,7 por cento dos pacientes do sexo masculino (n=174) e 25,3 por cento do sexo feminino (n=59). Em 63 exames (21,3 por cento) foram isolados fungos leveduriformes, sendo 49,2 por cento dos casos do sexo masculino (n=31) e 50,8 por cento do sexo feminino (n=32).As idades variaram entre 2 e 99 anos no grupo dos fungos filamentosos e entre 14 e 88 anos no grupo dos fungos leveduriformes. Os fungos filamentosos mais freqüentemente encontrados foram Fusarium (n=137 ou 58,8 por cento), Aspergillus (n=28 ou 12,0 por cento) e Penicillium (n=12 ou 5,2 por cento). Entre as leveduras, o gênero mais freqüente foi a Candida (n=59 ou 93,7 por cento). Em relação às ceratites, 95,1 por cento das solicitações apresentadas tiveram pelo menos um fator associado. Trauma ocular foi significativamente associado às ceratites por fungos filamentosos (p=0,0002); e, cirurgias (p=0,0002), doenças oculares prévias (p=0,0002) e sistêmicas (p=0,0002) às infecções por fungos leveduriformes. O uso de antibióticos tópicos mostrou associação com infecção por fungos filamentosos, com menor significância estatística (p=0,0224). CONCLUSÕES: A análise laboratorial mostrou predomínio de infecções por fungos filamentosos sobre os leveduriformes, destacando-se a ceratite micótica como a principal doença verificada. A maioria das ceratites fúngicas apresentou pelo menos um fator associado.

Comparaçäo do uso tópico de cetotifeno com a olopatadina no tratamento de conjuntivites alérgicas / Comparison between topical use of ketotifen and olopatadine in the treatment of allergic conjunctivitis

Objetivo: Avaliar e comparar a eficácia e tolerância do uso tópico do fumarato de cetotifeno a 0,05 por cento e cloridrato de olopatadina a 0,1 por cento no tratamento de pacientes com conjuntivite alérgica. Método: Foi realizado estudo clínico mascarado, randomizado comparando a eficácia, segurança e os efeitos colaterais com o uso da soluçäo oftálmica de fumarato de cetotifeno a 0,05 por cento e cloridrato de olopatadina a 0,1 por cento no alívio dos sintomas e sinais em pacientes com conjuntivite alérgica. Trinta e quatro pacientes obedecendo aos critérios de inclusäo do protocolo receberam um frasco com a droga mascarada e instilaram uma gota duas vezes por dia em cada olho durante 30 dias. Os sintomas e sinais dos pacientes foram avaliados em uma visita pré-tratamento e cinco com tratamento (1º dia, 2º dia, 7º dia, 14º dia e 30º dia). Resultados: A gravidade da conjuntivite alérgica foi semelhante nos dois grupos do estudo. Tanto cetotifeno como a olopatadina foram equivalentes e eficazes na diminuiçäo dos sintomas de prurido, ardor e lacrimejamento. Quanto aos sinais, a hiperemia em conjuntiva bulbar foi atenuada nos dois grupos. Na avaliaçäo das reaçöes adversas observou-se ardor após a administraçäo de ambos colírios e a ocorrência de prurido no grupo do cetotifeno. Näo foi observada nenhuma reaçäo de hipersensibilidade das drogas estudadas. Conclusöes: Este estudo evidencia que a soluçäo oftálmica de fumarato de cetotifeno a 0,05 por cento e o cloridrato de olopatadina a 0,1 por cento quando instilados duas vezes ao dia durante 30 dias, säo eficazes e seguros no alívio dos principais sintomas das conjuntivites alérgicas.

Análise laboratorial de ceratites fungicas em Serviço Universitário / Laboratorial analyses of fungal keratitis in a University Service

Apresentar a frequencia e o tipo de fungos identificados de infecçöes corneanas. Métodos: Levantamento retrospectivo dos casos de ceratites micóticas, no Laboratório Micribiologia Ocular do Departamento de Oftalmologia da Universidade Federal de Säo Paulo (UNIFESP) no período entre 1995 a 1998. Descriçäo dos isolamentos de fungos, análises dos fatores desencadeantes e relaçäo com o número de ceratites infecciosas no mesmo período. Resultados/Conclusäo: Ceratites micóticas foram diagnosticados em 61 (5,48 por cento) dos 113 pacientes que apresentaram úlcera de córnea de etiologia infecciosa, com variaçäo de 3,46 - 9,25 por cento, ao ano. Fungos filamentosos foram identificados em 47 casos (77,04 por cento) e leveduras em 14 (22,95 por cento). Fusarium foi o gênero mais frequente (50,82 por cento), seguido de Candida (22,95 por cento) e Aspergillus (8,19 por cento). Foram também isolados fungos raros com agentes etiológicos de ceratite como: Phaeosiaria sp; Phoma sp; Fonsecaea pedrosoi e Exserohilum rostratum.

Investigaçäo da micobiota conjuntival de indivíduos diabéticos, residentes em área urbana de Säo Paulo, Brasil / Mycobiota conjunctival investiagtion in diabetics who reside in urban area from Säo Paulo, Brasil

Objetivos: determinar a micobiota conjuntival em diabéticos residentes na zona urbana da cidade de São Paulo, segundo idade, sexo, tipo, tempo de diabetes e estádio da retinopatia diabética. Comparar com a micobiota conjuntival em indivíduos não diabéticos. Estabelecer a micobiota anemófila nas salas de colheita dos raspados de conjuntiva. Métodos: foi realizado estudo prospectivo, não aleatório com diabéticos em acompanhamento médico e indivíduos sem história prévia de diabetes melito. Este protocolo foi aprovado pelo comitê de ética desta universidade e obedeceu aos princípios da declaração de Helsique. Foram incluídas pessoas maiores de 18 anos, não usuárias de medicamentos tópicos ou lente de contato nos últimos 30 dias e não portadoras de doenças imunosupressoras ou doenças da superfície ocular. Os indivíduos avaliados foram submetidos a ectoscopia, raspado conjuntival bilateral, sem anestesia, utilizando zaragatoa alginatada esterilizada, embebida de salina esterilizada. Sabouraud dextrose ágar acrescido de cloranfenicol foi utilizado para primo-isolamento. Nos indivíduos diabéticos, foi realizada avaliação da retina e nos não diabéticos se aplicou um critério de triagem para diabetes não diagnosticada, sendo selecionadas pessoas com glicemia de sangue capilar < 110 mg. dl-1 (em jejum de 8 horas). A colheita dos fungos anemófilos da sala consistiu em expor placas com Sabouraud dextrose ágar acrescido de cloranfenicol por 30 minutos, durante as colheitas. Os cultivos foram observados a cada três dias, durante 30 dias, após a colheita. Para identificação dos fungos, foram utilizados exames macroscópicos e microscópicos das colônias suspeitas. A chave de identificação de De Hoog & Guarro foi adotada para fungos filamentosos. Para a determinação do tamanho da amostra dos diabéticos, foi considerada como proporção estimada de fungos em conjuntiva sadia 11,0 por cento, utilizando como nível de significância 5,0 por cento e intervalo de confiança 95,0 por cento. Para a análise dos dados, foram utilizadas técnicas de estatística descritiva e inferencial...(au)