SARS-CoV-2 and the ocular surface: test accuracy and viral load / SARS-CoV-2 e a superfície ocular: acurácia dos testes e carga viral

ABSTRACT Purpose: This study aimed to evaluate the pre-sence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in the ocular surface of individuals clinically suspected of coronavirus disease 2019 (COVID-19) and determine the accuracy of different approaches of molecular testing on the ocular surface based on the nasopharyngeal positivity status for COVID-19. Methods: A total of 152 individuals with suspected COVID-19 symptoms who simultaneously underwent nasopharyngeal and two different tear film collection techniques for quantitative reverse-transcriptase polymerase chain reaction (RT-qPCR) were included. Tears were collected and randomized: one eye had the filter strip for the Schirmer test and the contralateral eye had conjunctival swab/cytology in the inferior fornix. All patients underwent slit lamp biomicroscopy. The accuracy of various ocular surface collection techniques used for the detection of SARS-CoV-2 RNA was determined. Results: Of the 152 patients enrolled in the study, 86 (56.6%) had COVID-19 confirmed by nasopharyngeal PCR. Both tear film collection techniques detected viral particles: the Schirmer test was positive in 16.3% (14/86) and the conjunctival swab/cytology in 17.4% (15/86), with no statistically significant differences. No positive ocular tests were found among those with negative nasopharyngeal PCR tests. The overall agreement of the ocular tests was 92.7%, and in combination, the sensitivity would increase to 23.2%. The mean cycle threshold values in the nasopharyngeal, Schirmer, and conjunctival swab/cytology tests were 18.2 ± 5.3, 35.6 ± 1.4, and 36.4 ± 3.9, respectively. Compared with the nasopharyngeal test, the Schirmer (p=0.001) and conjunctival swab/cytology (p<0.001) tests had significantly different Ct values. Conclusion: The Schirmer (16.3%) and conjunctival swab (17.4%) tests were comparably capable of detecting SARS-CoV-2 RNA in the ocular surface by RT-PCR accurately based on nasopharyngeal status and demonstrated indistinct sensitivity and specificity. Simultaneous specimen sampling and processing from the nasopharyngeal, Schirmer, and conjunctival swab/cytology tests demonstrated significantly lower viral load in both ocular surface approaches than in the nasopharyngeal test. Ocular manifestations detected by slit lamp biomicroscopy were not associated with ocular RT-PCR positivity.

RESUMO Objetivo: Avaliar a presença de RNA de coronavírus 2 causador de síndrome respiratória aguda grave (SARS-CoV-2) na superfície ocular de indivíduos clinicamente suspeitos com COVID-19 e determinar a precisão de diferentes abordagens de testes moleculares na superfície ocular com base no status de positividade do RT-qPCR de nasofaringe para COVID-19. Métodos: 152 indivíduos com sintomas suspeitos para a COVID-19 foram submetidos a coleta de reação em cadeia da polimerase de nasofaringe simultaneamente a duas técnicas diferentes de coleta de filme lacrimal para RT-qPCR: aleatoriamente, um olho com a tira filtro do teste de Schirmer e, o olho contralateral, com citologia (swab) conjuntival no fórnice inferior. Todos os indivíduos foram submetidos à biomicroscopia com lâmpada de fenda. Resultados: Dos 152 pacientes, 86 (56,6%) tiveram a COVID-19 confirmada por PCR de nasofaringe. Ambas as técnicas de coleta detectaram partículas virais: o teste de Schirmer foi positivo em 16,3% (14/86) e a citologia conjuntival em 17,4% (15/86), sem diferenças estatisticamente significativas. Não houve testes oculares positivos entre aqueles com reação em cadeia da polimerase de nasofaringe negativo. A concordância geral dos testes oculares foi de 92,7% e, em combinação, a sensibilidade aumentaria para 23,2%. Os valores médios do limiar de ciclo nos testes de nasofaringe, Schirmer e citologia conjuntival foram 18,2 ± 5,3, 35,6 ± 1,4 e 36,4 ± 3,9, respectivamente. Conclusão: Os testes de Schirmer (16,3%) e swab conjuntival (17,4%) foram igualmente capazes de detectar RNA de SARS-CoV-2 na superfície ocular por RT-PCR e demonstraram sensibilidade e especificidade indistintas. A coleta simultânea de amostras ao processamento dos testes de RT-PCR de nasofaringe, Schirmer e citologia (swab) conjuntival demonstraram carga viral significativamente menor em ambas as abordagens da superfície ocular em comparação com o teste de nasofaringe. As manifestações oculares detectadas pela biomicroscopia com lâmpada de fenda não foram claramente associadas à positividade do RT-PCR ocular.

SARS-CoV-2 and the ocular surface: test accuracy and viral load.

PURPOSE: This study aimed to evaluate the pre-sence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA in the ocular surface of individuals clinically suspected of coronavirus disease 2019 (COVID-19) and determine the accuracy of different approaches of molecular testing on the ocular surface based on the nasopharyngeal positivity status for COVID-19. METHODS: A total of 152 individuals with suspected COVID-19 symptoms who simultaneously underwent nasopharyngeal and two different tear film collection techniques for quantitative reverse-transcriptase polymerase chain reaction (RT-qPCR) were included. Tears were collected and randomized: one eye had the filter strip for the Schirmer test and the contralateral eye had conjunctival swab/cytology in the inferior fornix. All patients underwent slit lamp biomicroscopy. The accuracy of various ocular surface collection techniques used for the detection of SARS-CoV-2 RNA was determined. RESULTS: Of the 152 patients enrolled in the study, 86 (56.6%) had COVID-19 confirmed by nasopharyngeal PCR. Both tear film collection techniques detected viral particles: the Schirmer test was positive in 16.3% (14/86) and the conjunctival swab/cytology in 17.4% (15/86), with no statistically significant differences. No positive ocular tests were found among those with negative nasopharyngeal PCR tests. The overall agreement of the ocular tests was 92.7%, and in combination, the sensitivity would increase to 23.2%. The mean cycle threshold values in the nasopharyngeal, Schirmer, and conjunctival swab/cytology tests were 18.2 ± 5.3, 35.6 ± 1.4, and 36.4 ± 3.9, respectively. Compared with the nasopharyngeal test, the Schirmer (p=0.001) and conjunctival swab/cytology (p<0.001) tests had significantly different Ct values. CONCLUSION: The Schirmer (16.3%) and conjunctival swab (17.4%) tests were comparably capable of detecting SARS-CoV-2 RNA in the ocular surface by RT-PCR accurately based on nasopharyngeal status and demonstrated indistinct sensitivity and specificity. Simultaneous specimen sampling and processing from the nasopharyngeal, Schirmer, and conjunctival swab/cytology tests demonstrated significantly lower viral load in both ocular surface approaches than in the nasopharyngeal test. Ocular manifestations detected by slit lamp biomicroscopy were not associated with ocular RT-PCR positivity.

B-scan ultrasound, visual electrophysiology and perioperative videoendoscopy for predicting functional results in keratoprosthesis candidates.

BACKGROUND/AIMS: We analysed the ability of B-scan ultrasound, ocular electrophysiology testing and videoendoscopic examination for predicting visual prognosis in Boston Type 1 keratoprosthesis (KPro-1) candidates. Indirect anatomical and electrophysiological findings and results from direct endoscopic evaluations were correlated with postoperative functional data. METHODS: In this prospective and interventional study, we included 13 individuals who had previously been indicated for Kpro-1 surgery. All subjects underwent preoperative screening, including ophthalmic evaluation, B-scan ultrasound, electrophysiological testing, and perioperative intraocular videoendoscopic evaluation (VE). B-scan ultrasound, electrophysiological testing, and VE evaluation results were categorised as favourable or unfavourable predictors of postoperative functional results according to predefined criteria. The predictability values of B-scan ultrasound, electrophysiological testing, and VE prognostication were calculated based on the visual acuity level achieved. RESULTS: All surgeries and perioperative VEs were uneventful. Preoperative best-corrected visual acuity (BCVA) ranged from light perception to counting fingers. The 1-year postoperative BCVA was better than 20/200 (satisfactory visual acuity result) in 10 eyes (76.9%) and 20/40 or better in 5 eyes (38.5%). B-scan ultrasound presented a positive predictive value (PPV) of 85.7% for satisfactory postoperative visual acuity, electroretinography showed a PPV of 66.7%, and visual evoked potential presented a PPV of 66.7%. The perioperative VE PPV of a negative finding for satisfactory visual acuity was 100%. CONCLUSIONS: Fundoscopic visualisation by intraocular VE is a minimally invasive procedure that can be used to predict functional outcomes in keratoprosthesis candidates. This technique demonstrated better prognostication in keratoprosthesis candidates than B-scan ultrasound and electrophysiological testing.

Fungal keratitis management in a referral cornea center in Brazil / Manejo de ceratites fúngicas em centro de referência no Brasil

Abstract Purpose: To report etiological diagnosis, predisposing risk factors, therapeutic strategies and visual outcome of patients treated at the Department of Ophthalmology of Federal University of São Paulo. Methods: This is a retrospective, descriptive, and observational study from medical and laboratory records of the Department of Ophthalmology of Federal University of São Paulo, including all patients with culture proven fungal keratitis in 5 years, from October 2012 through October 2017. Results: There were 2260 fungi microbiologic test requests. Of these, 140 samples had positive cultures for fungi and sixty-six patients were followed at our clinic. Forty-five patients (68.2%) were men, and the mean age was 48.06 (±17.39) years. Fusarium spp. was the most frequently isolated fungus (32 cases; 48.5%), followed by Candida parapsilosis (12 cases; 18.2%). Thirty-four patients (51.5%) underwent intracameral injection of amphotericin B (5 µg per 0.1 ml). In 11 patients (32.3%), infection was eradicated after intracameral amphotericin B associated to topical antifungal treatment and, in 23 patients (67.7%), therapeutic keratoplasty was needed. No complication related to intracameral amphotericin B injection was observed in this series. Forty-three patients (65.1%) ended up with therapeutic keratoplasty. Three patients (4.5%) evolved to evisceration or enucleation. At the last follow-up visit, 53 patients (80.3%) had visual acuity worse than 20/200. Conclusion: Despite current antifungals drugs and distinct administration strategies, fungal keratitis remains challenging. Delayed antifungal therapy may explain poor clinical outcomes. Intracameral amphotericin B associated to topical antfungal treatment seems to be a safe and helpful alternative for non-responsive fungal keratitis. But it is important to formulate other treatment strategies, hence to improve patients' outcomes, since most patients ended-up with significant visual impairment even after current treatment.

Resumo Objetivo: Descrever diagnósticos etiológicos, fatores de risco, estratégias terapêuticas e resultados visuais de pacientes com ceratite fúngica tratados no Departamento de Oftalmologia da Universidade Federal de São Paulo. Métodos: Trata-se de um estudo retrospectivo, descritivo e observacional, a partir da análise de prontuários médicos e laboratoriais do Departamento de Oftalmologia da Universidade Federal de São Paulo, incluindo todos os pacientes com ceratite fúngica comprovada por cultura no período de outubro de 2012 a outubro de 2017. Resultados: Foram realizadas 2260 solicitações de testes microbiológicos. Destas, 140 amostras apresentaram culturas positivas para fungos, e 66 pacientes foram acompanhados em nosso serviço. Quarenta e cinco pacientes (68,2%) eram do sexo masculino, e a média de idade foi de 48,06 (± 17,39) anos. Fusarium spp. foi o fungo mais freqüentemente isolado (32 casos; 48,5%), seguido por Candida parapsilosis (12 casos; 18,2%). Trinta e quatro pacientes (51,5%) foram submetidos à injeção intracameral de anfotericina B (5 µg por 0,1 ml). Destes, 11 pacientes (32,3%) tiveram a infecção erradicada. Nos outros 23 pacientes (67,7%), o transplante terapêutico foi necessário. Nenhuma complicação relacionada à injeção intracameral de anfotericina B foi observada neste estudo. No total, 43 pacientes (65,1%) evoluíram para transplante terapêutico, e 3 pacientes (4,5%) foram submetidos à evisceração ou enucleação. Cinquenta e três pacientes (80,3%) apresentaram acuidade visual final pior que 20/200. Conclusões: Apesar dos diversos medicamentos antifúngicos atuais e vias de administração, o tratamento das ceratites fúngicas permanece desafiador. O atraso no início do tratamento adequado pode justificar o desfecho clínico desfavorável de grande parte dos pacientes. A injeção intracameral de anfotericina B mostrou-se uma alternativa terapêutica segura para ceratites fúngicas refratárias. Mas outras estratégias de tratamento devem ser formuladas, visando melhorar os resultados visuais dos pacientes.

Long-term outcomes with Boston type 1 keratoprosthesis in ocular burns.

PURPOSE: To evaluate the long-term outcomes of Boston type I keratoprosthesis (B-Kpro) in the management of patients with ocular burn injuries. METHODS: This prospective study included all patients with ocular burns who underwent B-Kpro implantation at a tertiary eye care center between February 2008 and November 2015. Twelve patients (12 eyes) were enrolled. The procedures performed for managing ocular injury were identified, and information on ocular history, surgical procedures performed, and postoperative outcomes was collected. The main outcome measures were visual acuity, prosthesis retention, postoperative complications, and required surgical procedures. RESULTS: Twelve eyes from 12 patients met the inclusion criteria for B-Kpro implantation, including nine eyes with alkali burns and three eyes with thermal burns. A total of 13 B-Kpro devices were implanted in the 12 eyes. The mean follow-up period was 60.8 months (range, 13-91 months). Preoperative best-corrected visual acuity ranged from counting fingers to light perception. Postoperative best-corrected visual acuity was better than 20/200 in 83.3%, 66.6%, and 57.1% of patients at 12, 48, and 60 months, respectively. The initial keratoprosthesis was retained in 10 (83.3%) eyes and was successfully replaced in one eye. The major cause of worsening of initial visual acuity was advanced glaucoma (four of 12 eyes). CONCLUSION: The anatomical and functional results support the use of B-Kpro for managing bilateral limbal stem cell deficiency secondary to ocular burns. However, glaucoma should be carefully evaluated, as it is a continuous threat that may result in irreversible visual loss in this population.

Long-term outcomes with Boston type 1 keratoprosthesis in ocular burns / Resultados a longo prazo com o implante da ceratoprótese de Boston tipo 1 em queimaduras oculares

ABSTRACT Purpose: To evaluate the long-term outcomes of Boston type I keratoprosthesis (B-Kpro) in the management of patients with ocular burn injuries. Methods: This prospective study included all patients with ocular burns who underwent B-Kpro implantation at a tertiary eye care center between February 2008 and November 2015. Twelve patients (12 eyes) were enrolled. The procedures performed for managing ocular injury were identified, and information on ocular history, surgical procedures performed, and postoperative outcomes was collected. The main outcome measures were visual acuity, prosthesis retention, postoperative complications, and required surgical procedures. Results: Twelve eyes from 12 patients met the inclusion criteria for B-Kpro implantation, including nine eyes with alkali burns and three eyes with thermal burns. A total of 13 B-Kpro devices were implanted in the 12 eyes. The mean follow-up period was 60.8 months (range, 13-91 months). Preoperative best-corrected visual acuity ranged from counting fingers to light perception. Postoperative best-corrected visual acuity was better than 20/200 in 83.3%, 66.6%, and 57.1% of patients at 12, 48, and 60 months, respectively. The initial keratoprosthesis was retained in 10 (83.3%) eyes and was successfully replaced in one eye. The major cause of worsening of initial visual acuity was advanced glaucoma (four of 12 eyes). Conclusion: The anatomical and functional results support the use of B-Kpro for managing bilateral limbal stem cell deficiency secondary to ocular burns. However, glaucoma should be carefully evaluated, as it is a continuous threat that may result in irreversible visual loss in this population.

RESUMO Objetivo: Avaliar os resultados em longo prazo do implante da ceratoprótese de Boston tipo 1 (B-Kpro) no tratamento de pacientes vítima de queimadura ocular. Métodos: Trata-se de um estudo prospectivo envolvendo todos os casos de implante de B-Kpro 1 para queimadura ocular em um centro de oftalmologia terciário durante o período de fevereiro/2008 e novembro/2015. Doze pacientes (doze olhos) foram incluídos no estudo. Os procedimentos realizados para o manejo da queimadura ocular foram identificados, e os dados foram coletados com relação à história oftalmológica do paciente, procedimentos cirúrgicos realizados, e resultados pós-operatórios. Os principais parâmetros avaliados foram acuidade visual, retenção da prótese, complicações pós-ope ratórias e intervenções cirúrgicas necessárias. Resultados: Doze pacientes preencheram os critérios de inclusão para implante da B-Kpro, incluindo 09 olhos vítimas de queimadura por álcali e 03 olhos pós queimadura térmica. Um total de 13 B-Kpro tipo 1 foram implantadas em 12 olhos de 12 pacientes. O tempo de seguimento médio foi de 60,8 meses (variando, 13-91 meses). Melhor acuidade visual corrigida pré-operatória variou de conta dedos à percepção luminosa. Melhor acuidade visual corrigida pós-operatória foi melhor que 20/200 em 83,3%, 66,6% e 57,1% dos pacientes em 12 meses, 48 meses e 60 meses, respectivamente. A ceratoprótese inicialmente implantada permaneceu retida em 10 (83,3%) dos olhos e foi re-implantada com sucesso em um olho. A principal causa de piora da acuidade visual inicialmente atingida foi glaucoma avançado (4 de 12 olhos). Conclusão: Os resultados anatômicos e funcionais reportados neste estudo dão suporte ao uso da B-Kpro para o tratamento da deficiência límbica bilateral secundária a queimadura ocular. Glaucoma é o principal fator limitante e pode causar perda visual irreversível nesses pacientes.

Anterior segment optical coherence tomography findings in type I Boston keratoprosthesis.

PURPOSE: To report the results of high-resolution anterior segment optical coherence tomography of patients implanted with a type 1 Boston keratoprosthesis (KPro). METHODS: The retrospective study cohort included 11 eyes of 11 patients (average age, 58.4 years; range, 34-83 years). All subjects underwent anterior segment optical coherence tomography at a single posteoperative time point. The main outcome measures were retro-backplate and retro-optic membrane formation, thinning and gap formation of the corneal carrier graft (melting), and degree of angle closure. RESULTS: Preoperative diagnoses included chemical burn (55%), failed corneal transplant (36%), and Stevens-Johnson syndrome (9%). The mean postoperative follow-up duration was 38.5 (range, 12-72) months. The most frequent findings of anterior segment optical coherence tomography were retroprosthetic membrane formation (63%, 7/11), thinning of the corneal carrier graft (melting; 55%, 6/11), and a narrow or closed angle (91%, 10/11). Other less common findings were epithelial growth over the optic surface and periprosthetic cyst formation. Retroprosthetic membrane formation was observed in all patients with melting (6/11). CONCLUSIONS: Detailed postoperative examination and visualization of subtle changes of keratoprosthesis implanted eyes by slit lamp biomicroscopy are often difficult. Anterior segment optical coherence tomography is a useful, noninvasive, and quantitative imaging technique that provides useful information to postoperatively monitor the anatomic stability of an implanted keratoprosthesis.

Anterior segment optical coherence tomography findings in type I Boston keratoprosthesis / Achados de tomografia de coerência óptica do seguimento anterior em ceratoprótese de Boston tipo I

ABSTRACT Purpose: To report the results of high-resolution anterior segment optical coherence tomography of patients implanted with a type 1 Boston keratoprosthesis (KPro). Methods: The retrospective study cohort included 11 eyes of 11 patients (average age, 58.4 years; range, 34-83 years). All subjects underwent anterior segment optical coherence tomography at a single posteoperative time point. The main outcome measures were retro-backplate and retro-optic membrane formation, thinning and gap formation of the corneal carrier graft (melting), and degree of angle closure. Results: Preoperative diagnoses included chemical burn (55%), failed corneal transplant (36%), and Stevens-Johnson syndrome (9%). The mean postoperative follow-up duration was 38.5 (range, 12-72) months. The most frequent findings of anterior segment optical coherence tomography were retroprosthetic membrane formation (63%, 7/11), thinning of the corneal carrier graft (melting; 55%, 6/11), and a narrow or closed angle (91%, 10/11). Other less common findings were epithelial growth over the optic surface and periprosthetic cyst formation. Retroprosthetic membrane formation was observed in all patients with melting (6/11). Conclusions: Detailed postoperative examination and visualization of subtle changes of keratoprosthesis implanted eyes by slit lamp biomicroscopy are often difficult. Anterior segment optical coherence tomography is a useful, noninvasive, and quantitative imaging technique that provides useful information to postoperatively monitor the anatomic stability of an implanted keratoprosthesis.

RESUMO Objetivos: Reportar os resultados das imagens de pacientes com Ceratoprótese de Boston tipo I (KPro) usando tomografia de coerência óptica de alta resolução do seguimento anterior (AS-OCT). Métodos: Nós realizamos um estudo retrospectivo de pacientes submetidos à KPro. Um total de 11 olhos de 11 pacientes foram incluídos. As imagens de AS-OCT foram realizadas em um único tempo de pós-operatório. Os principais resultados incluem formação de membrana retroprostética atrás do prato posterior e atrás do cilindro ótico, afinamento e lacunas na córnea doadora (melt) e graus de ângulo fechado. Resultados: Os diagnósticos pré-operatórios inclui queimadura química (55%), falência pós transplante de córnea (36%) e síndrome de Stevens Johnson (9%). A idade média foi de 58.4 anos (escala, 34-83 anos). A média de tempo de pós-operatório foi de 38.5 meses (escala, 12-72 meses). Os achados mais frequentes de AS-OCT foram: membrana retroprostética, 63% (7/11); afinamento da córnea doadora (melting), 55% (6/11); angulo estreito ou fechado, 91% (10/11). Outros achados menos comuns foram crescimento epitelial sobre a superfície ótica e cistos periprostéticos. Todos os pacientes com melting (6/11) apresentaram membrana retroprostética. Conclusões: O exame pós-operatório e a visualização detalhada das mudanças em olhos com KPro pela lâmpada de fenda pode ser difícil. AS-OCT é uma técnica de imagem útil, não invasiva e quantitativa que permite o monitoramento da estabilidade anatômica no seguimento de KPro implantadas.

Limbal transplantation at a tertiary hospital in Brazil: a retrospective study.

PURPOSE: Epidemiological analysis of limbal transplantation surgeries performed in the Ophthalmologic Hospital of Sorocaba. METHODS: Retrospective medical records review of 30 patients who underwent limbal stem cell transplants between January 2003 and March 2008. Cases involving conjunctival limbal autograft were classified as group I, and those involving conjunctival limbal allograft as group II. RESULTS: Two patients were excluded due to incomplete data during postoperative follow-up. Of the total sample of 28 patients, 53.6% constituted group I, whereas 46.4% were included in group II. Males were predominant (67.9%), and right eyes were the most prevalent (67.9%). The mean age was 40.3 years. Unilateral cases accounted for 60.7%. The most frequent pathology causing limbal system failure was chemical burns (53%). The mean length of time from diagnosis to surgery was 11.18 years. The limbal graft and amniotic membrane were associated in 75% of all cases, and tarsorrhaphy in 57.1%. The average follow-up period was 24.84 months. The uncorrected visual acuity improved in 38% of the cases, was unchanged in 28.5%, and deteriorated in 33.3%. There was no persistent epithelial defect in 75% of the patients. The conjunctivalization rate was similar between the groups (53.3% and 58.3%, respectively). The transparency improved in only 38.4% of the cases, and 28.5% of the surgeries performed were successful. The most prevalent complication was persistent epithelial defect, which occurred in 25% of the patients, followed by corneal melting in 14.2%. Other complications observed included infectious ulcers, limbal graft necrosis or ischemia, perforation, and descemetocele. CONCLUSION: Chemical burns remain the main cause of limbal stem cell deficiency. In these cases, limbal transplantation is the standard procedure to restore the ocular surface even though the success rate is low.

Limbal transplantation at a tertiary hospital in Brazil: a retrospective study / Transplante de limbo em hospital terciário no Brasil: estudo retrospectivo

ABSTRACTPurpose:Epidemiological analysis of limbal transplantation surgeries performed in the Ophthalmologic Hospital of Sorocaba.Methods:Retrospective medical records review of 30 patients who underwent limbal stem cell transplants between January 2003 and March 2008. Cases involving conjunctival limbal autograft were classified as group I, and those involving conjunctival limbal allograft as group II.Results:Two patients were excluded due to incomplete data during postoperative follow-up. Of the total sample of 28 patients, 53.6% constituted group I, whereas 46.4% were included in group II. Males were predominant (67.9%), and right eyes were the most prevalent (67.9%). The mean age was 40.3 years. Unilateral cases accounted for 60.7%. The most frequent pathology causing limbal system failure was chemical burns (53%). The mean length of time from diagnosis to surgery was 11.18 years. The limbal graft and amniotic membrane were associated in 75% of all cases, and tarsorrhaphy in 57.1%. The average follow-up period was 24.84 months. The uncorrected visual acuity improved in 38% of the cases, was unchanged in 28.5%, and deteriorated in 33.3%. There was no persistent epithelial defect in 75% of the patients. The conjunctivalization rate was similar between the groups (53.3% and 58.3%, respectively). The transparency improved in only 38.4% of the cases, and 28.5% of the surgeries performed were successful. The most prevalent complication was persistent epithelial defect, which occurred in 25% of the patients, followed by corneal melting in 14.2%. Other complications observed included infectious ulcers, limbal graft necrosis or ischemia, perforation, and descemetocele.Conclusion:Chemical burns remain the main cause of limbal stem cell deficiency. In these cases, limbal transplantation is the standard procedure to restore the ocular surface even though the success rate is low.

RESUMOObjetivo:Análise epidemiológica dos transplantes de limbo realizados no Hospital Oftalmológico de Sorocaba.Método:Foi realizada uma análise retrospectiva através de revisão de prontuário de trinta casos de transplante de limbo realizados no período de janeiro de 2003 a março de 2008. Casos de transplante de limbo autólogo foram classificados como grupo I e alogênicos como grupo II.Resultados:Dois pacientes foram excluídos da análise por insuficiência de dados nos registros. Da amostra completa de 28 pacientes, 53,6% constituíram o grupo I enquanto 46,4% o grupo II. O olho direito (68%) e o sexo masculino (68%) foram mais acometidos, com uma média de idade de 40,3 anos. Casos unilaterais contabilizaram 60,7%. A patologia de base causadora da deficiência límbica mais prevalente foi a queimadura química (53%). A média do tempo de doença até a cirurgia foi de 11,18 anos. Na maioria dos casos o transplante foi associado a membrana amniótica (75%) e tarsorrafia (57%). O tempo médio de seguimento foi de 24,84 meses. Foi observado melhora da acuidade visual não corrigida em 38% dos casos enquanto 28,5% permaneceram inalteradas e 33,3% pioraram. Evolução sem defeito epitelial persistente ocorreu em 75% dos pacientes. A taxa de conjuntivalização foi semelhante nos 2 grupos (53,3% e 58,3%). Em apenas 38% dos casos houve melhora da transparência. A taxa de sucesso foi de cerca de 28%. A complicação mais prevalente foi defeito epitelial persistente (25%) seguida de melting (14,2%). Outras complicações observadas foram úlceras infecciosas, necrose ou isquemia do enxerto, perfuração e descemetocele.Conclusões:A queimadura química permanece como principal patologia causadora de deficiência límbica. Nestes casos o transplante de limbo é atualmente a técnica de eleição para restauração da superfície ocular, porém com baixa taxa de sucesso.

Boston type I keratoprosthesis: Review.

Regardless of significant progress in the field of corneal transplantation to treat corneal opacification, some cases of corneal blindness still present a poor prognosis for conventional penetrating keratoplasty. In patients with repeated graft failure and/or with severe ocular surface disease, the Boston type I keratoprosthesis (type I BKPro) has become a viable option. Modifications in its design and postoperative management have improved the long-term outcomes of visual acuity, retention, and postoperative infection rates. These advances made the type I BKPro be considered a safe alternative for visual rehabilitation in many patients with corneal pathologies. However, postoperative handle of chronic comorbidities, such as glaucoma, is still critical for preserving the visual gains achieved with BKPro.

Boston type I keratoprosthesis: Review / Ceratoprótese de Boston tipo I: Revisão

Regardless of significant progress in the field of corneal transplantation to treat corneal opacification, some cases of corneal blindness still present a poor prognosis for conventional penetrating keratoplasty. In patients with repeated graft failure and/or with severe ocular surface disease, the Boston type I keratoprosthesis (type I BKPro) has become a viable option. Modifications in its design and postoperative management have improved the long-term outcomes of visual acuity, retention, and postoperative infection rates. These advances made the type I BKPro be considered a safe alternative for visual rehabilitation in many patients with corneal pathologies. However, postoperative handle of chronic comorbidities, such as glaucoma, is still critical for preserving the visual gains achieved with BKPro.

Apesar dos avanços consideráveis nas técnicas de transplantes da córnea para o tratamento de opacidades corneanas, alguns casos de cegueira de etiologia corneana ainda apresentam prognóstico reservado com a ceratoplastia penetrante convencional. Nesses pacientes, tais como aqueles vitimas de múltiplas falências após transplante de córnea e/ou com doença da superfície ocular avançada, a ceratoprótese de Boston tipo I (BKPro tipo I) tem se tornado uma opção viável. Modificações no seu modelo e no manejo pós-operatório melhoraram os resultados de acuidade visual, retenção e taxas de infecção pós-operatória a longo prazo. Devido às melhorias no dispositivo, a BKPro tipo I tem sido considerada uma alternativa cirúrgica segura para a reabilitação visual em muitos pacientes com patologias corneanas avançadas. Contudo, o controle pós-operatório de co-morbidades crônicas, como glaucoma, é fundamental para preservar os ganhos visuais obtidos com a BKPro.

Infectious keratitis in patients undergoing Boston Type 1 keratoprosthesis (Boston KPro) procedure: case series.

Description of two cases of infectious keratitis in patients after Boston Type 1 keratoprosthesis (Boston KPro) implantation. The first case refers to a patient that had the device indicated due to limbal deficiency secondary to severe dry eye who presented a fungal infection by Aerobasidium pullulans that was successfully treated with amphotericin B eye drops. The second case reports a patient with Boston KPro implantation due to previous corneal transplant rejection showing bacterial keratitis in the fourth postoperative month. The etiologic agent was identified as Streptococcus sp and topical treatment with vancomycin was effective. The importance of postoperative surveillance in Boston KPro eyes is discussed.

Infectious keratitis in patients undergoing Boston Type 1 keratoprosthesis (Boston KPro) procedure: case series / Ceratite infecciosa em pacientes submetidos ao implante de ceratoprótese Boston Tipo I: série de casos

Description of two cases of infectious keratitis in patients after Boston Type 1 keratoprosthesis (Boston KPro) implantation. The first case refers to a patient that had the device indicated due to limbal deficiency secondary to severe dry eye who presented a fungal infection by Aerobasidium pullulans that was successfully treated with amphotericin B eye drops. The second case reports a patient with Boston KPro implantation due to previous corneal transplant rejection showing bacterial keratitis in the fourth postoperative month. The etiologic agent was identified asStreptococcus sp and topical treatment with vancomycin was effective. The importance of postoperative surveillance in Boston KPro eyes is discussed.

Descrição de dois casos de ceratite infecciosa em pacientes submetidos a implante de ceratoprótese Boston Tipo 1 (Boston KPro). O primeiro caso refere-se a uma paciente na qual o dispositivo foi indicado por deficiência límbica secundária a olho seco grave, no qual foi identificado infecção fúngica por aerobasidium pullulans, tratada com sucesso com colírio de anfotericina B. O segundo caso reporta uma paciente com implante de Boston KPro por falências de transplantes de córnea prévios, que apresentou ceratite bacteriana no quarto mês pós-operatório. O agente etiológico identificado foi Streptococcus sp e o tratamento tópico com vancomicina foi eficaz. Discute-se a importância da vigilância pós-operatória em olhos submetidos ao implante de ceratoprótese.

Gene transfer to primary corneal epithelial cells with an integrating lentiviral vector / Transferência de genes heterólogos para células corneanas epiteliais primárias utilizando vetor lentivírus

PURPOSE: To evaluate the transfer of heterologous genes carrying a Green Fluorescent Protein (GFP) reporter cassette to primary corneal epithelial cells ex vivo. METHODS: Freshly enucleated rabbit corneoscleral tissue was used to obtain corneal epithelial cell suspension via enzymatic digestion. Cells were plated at a density of 5×10³ cells/cm² and allowed to grow for 5 days (to 70-80 percent confluency) prior to transduction. Gene transfer was monitored using fluorescence microscopy and fluorescence activated cell sorter (FACS). We evaluated the transduction efficiency (TE) over time and the dose-response effect of different lentiviral particles. One set of cells were dual sorted by fluorescence activated cell sorter for green fluorescent protein expression as well as Hoechst dye exclusion to evaluate the transduction of potentially corneal epithelial stem cells (side-population phenotypic cells). RESULTS: Green fluorescent protein expressing lentiviral vectors were able to effectively transduce rabbit primary epithelial cells cultured ex vivo. Live cell imaging post-transduction demonstrated GFP-positive cells with normal epithelial cell morphology and growth. The transduction efficiency over time was higher at the 5th post-transduction day (14.1 percent) and tended to stabilize after the 8th day. The number of transduced cells was dose-dependent, and at the highest lentivirus concentrations approached 7 percent. When double sorted by fluorescence activated cell sorter to isolate both green fluorescent protein positive and side population cells, transduced side population cells were identified. CONCLUSIONS: Lentiviral vectors can effectively transfer heterologous genes to primary corneal epithelial cells expanded ex vivo. Genes were stably expressed over time, transferred in a dose-dependence fashion, and could be transferred to mature corneal cells as well as presumable putative stem cells.

OBJETIVO: Avaliar a transferência de genes heterólogos expressando a proteí­na "Green Fluorescent Protein" (GFP) para células corneanas epiteliais primárias ex vivo utilizando vetor lentivírus. MÉTODOS: Tecido corneoescleral de coelhos foi usado para obtenção de suspensão de células corneanas epitelias. As células foram semeadas na densidade de 5×10³ células/cm² e expandidas por 5 dias até uma confluência de 70-80 por cento antes de serem transduzidas. A transferência genética foi monitorada por microscopia fluorescente e por um separador de células ativadas por fluorescência. Foram avaliadas a eficiência de transdução ao longo do tempo e o efeito dose-resposta de diferentes quantidades de partículas virais. Um grupo de células foi analisado pelo separador de células ativadas por fluorescência para avaliar a transdução de células com fenótipo de cé­lulas tronco do epitélio corneano (baseado na exclusão do corante "Hoechst dye"). RESULTADOS: Os vetores lentivírus foram efetivos na transdução de cé­lulas corneanas epiteliais primárias de coelhos ex vivo. Fotodocumentação das células vivas demonstrou células epiteliais de morfologia normal e expressando o gene fluorescente (GFP). A eficiência de transdução ao longo do tempo foi maior no quinto dia após a transdução (14,1 por cento) e demonstrou uma tendência à estabilidade a partir do oitavo dia após a transdução. O número de células transduzidas foi dose-dependente e atingiu 7 por cento com as maiores concentrações de partículas virais. Quando analisadas pelo separador de células ativadas por fluorescência para detecção de células transduzidas e também de células que excluíram o corante "Hoechst dye", foi detectado que células com fenótipo de células tronco do epitélio corneano ("side-population") também foram transduzidas. CONCLUSÕES: Os vetores lentivirais podem transferir genes heterolólogos para células corneanas epiteliais primárias expandidas ex vivo de forma eficiente. Os genes foram expressos de forma estável ao longo do tempo e puderam ser transferidos tanto para células epiteliais maduras como para presumíveis células tronco epiteliais. A eficiência de transdução foi obtida de forma dose-dependente.

Gene transfer to primary corneal epithelial cells with an integrating lentiviral vector.

PURPOSE: To evaluate the transfer of heterologous genes carrying a Green Fluorescent Protein (GFP) reporter cassette to primary corneal epithelial cells ex vivo. METHODS: Freshly enucleated rabbit corneoscleral tissue was used to obtain corneal epithelial cell suspension via enzymatic digestion. Cells were plated at a density of 5×10³ cells/cm² and allowed to grow for 5 days (to 70-80% confluency) prior to transduction. Gene transfer was monitored using fluorescence microscopy and fluorescence activated cell sorter (FACS). We evaluated the transduction efficiency (TE) over time and the dose-response effect of different lentiviral particles. One set of cells were dual sorted by fluorescence activated cell sorter for green fluorescent protein expression as well as Hoechst dye exclusion to evaluate the transduction of potentially corneal epithelial stem cells (side-population phenotypic cells). RESULTS: Green fluorescent protein expressing lentiviral vectors were able to effectively transduce rabbit primary epithelial cells cultured ex vivo. Live cell imaging post-transduction demonstrated GFP-positive cells with normal epithelial cell morphology and growth. The transduction efficiency over time was higher at the 5(th) post-transduction day (14.1%) and tended to stabilize after the 8(th) day. The number of transduced cells was dose-dependent, and at the highest lentivirus concentrations approached 7%. When double sorted by fluorescence activated cell sorter to isolate both green fluorescent protein positive and side population cells, transduced side population cells were identified. CONCLUSIONS: Lentiviral vectors can effectively transfer heterologous genes to primary corneal epithelial cells expanded ex vivo. Genes were stably expressed over time, transferred in a dose-dependence fashion, and could be transferred to mature corneal cells as well as presumable putative stem cells.

Effect of topical 0.5 percent povidone-iodine compared to 5 percent natamycin in fungal keratitis caused by Fusarium solani in a rabbit model: a pilot study / Efeito tópico de iodo-povidona 0, 5 por cento comparado com natamicina 5 por cento em ceratites fúngicas causadas por Fusarium solani em um modelo animal de coelhos: estudo piloto

PURPOSE: To evaluate the efficacy of topical administration of 0.5 percent povidone-iodine in experimental Fusarium solani keratitis in rabbits. METHODS: Fungal keratitis caused by Fusarium solani was induced in the right eye of 24 New Zealand rabbits. The rabbits were randomly divided into 3 different treatment groups: Group I (povidone-iodine) - treated with topical 0.5 percent povidone-iodine; Group II (natamycin) - treated with topical 5 percent natamycin; and Group III (control) - treated with topical saline solution. In all groups the rabbits were treated for three days and then sacrificed. The corneas were excised, macerated and immersed in 10 mL BHI. Culture samples were plated daily on Sabouraud's agar for 7 days, and the number of colony-forming units (CFU) was counted. The rabbits were clinically evaluated during the treatment period. RESULTS: The povidone-iodine and natamycin groups demonstrated better efficacy than the control group based on the number of rabbits with no colonies growing. However, there were no statistically significant differences between the three groups when the number of CFU was analyzed (p>0.05). CONCLUSIONS: Our study demonstrates important methodological considerations in the use of in vivo animal models for the testing of antifungal agents. Using this sample size and methodology of counting CFU, topical 0.5 percent povidone-iodine demonstrated no benefit in the treatment of experimental Fusarium solani when compared with topical 5 percent natamycin.

OBJETIVO: Avaliar a eficácia do uso tópico de iodo-povidona 0,5 por cento em ceratite experimental por Fusarium solani em coelhos. MÉTODOS: Ceratite fúngica por Fusarium solani foi induzida no olho direito de 24 coelhos da raça New Zealand. Os coelhos foram divididos aleatoriamente em 3 diferentes grupos de tratamento: Grupo I (iodo-povidona) - tratados com iodo-povidona 0,5 por cento; Grupo II (natamicina) - tratados com natamicina 5 por cento; Grupo III (controle) - tratados com solução salina. Os coelhos dos 3 grupos foram tratados por 3 dias e sacrificados em seguida. As córneas foram removidas cirurgicamente, maceradas e incubadas em meio BHI. Semeou-se culturas em placas de ágar Sabouraud, diariamente, durante 7 dias, e contou-se o número de unidades formadoras de colônias (UFC). Os coelhos foram avaliados clinicamente durante o período de tratamento. RESULTADOS: Os grupos iodo-povidona e natamicina demonstraram melhor eficácia do que o grupo controle considerando-se o número de coelhos nos quais não houve crescimento de colônias. Entretanto, não houve diferença estatística significante entre os 3 grupos quando se analizou o número de UFC (p>0,05). CONCLUSÃO: Este estudo demonstrou considerações metodológicas importantes na utilização de modelos animais para o teste de agentes antifúngicos. Usando a metodologia de contar UFC e com este tamanho amostral, administração tópica de iodo-povidona 0,5 por cento não demonstrou benefício do tratamento de ceratite fúngica experimental causada por Fusarium solani quando comparado com a administração tópica de natamicina 5 por cento.

Effect of topical 0.5% povidone-iodine compared to 5% natamycin in fungal keratitis caused by Fusarium solani in a rabbit model: a pilot study.

PURPOSE: To evaluate the efficacy of topical administration of 0.5% povidone-iodine in experimental Fusarium solani keratitis in rabbits. METHODS: Fungal keratitis caused by Fusarium solani was induced in the right eye of 24 New Zealand rabbits. The rabbits were randomly divided into 3 different treatment groups: Group I (povidone-iodine) - treated with topical 0.5% povidone-iodine; Group II (natamycin) - treated with topical 5% natamycin; and Group III (control) - treated with topical saline solution. In all groups the rabbits were treated for three days and then sacrificed. The corneas were excised, macerated and immersed in 10 mL BHI. Culture samples were plated daily on Sabouraud's agar for 7 days, and the number of colony-forming units (CFU) was counted. The rabbits were clinically evaluated during the treatment period. RESULTS: The povidone-iodine and natamycin groups demonstrated better efficacy than the control group based on the number of rabbits with no colonies growing. However, there were no statistically significant differences between the three groups when the number of CFU was analyzed (p>0.05). CONCLUSIONS: Our study demonstrates important methodological considerations in the use of in vivo animal models for the testing of antifungal agents. Using this sample size and methodology of counting CFU, topical 0.5% povidone-iodine demonstrated no benefit in the treatment of experimental Fusarium solani when compared with topical 5% natamycin.

Are cutaneous hypersensitivity tests to inhalant allergens a severity marker for vernal keratoconjunctivitis?

PURPOSE: The purpose of this study was to analyze the cutaneous sensitivity to a variety of allergens in patients with vernal keratoconjunctivitis (VKC) and to demonstrate the relation between skin response and clinical aspects of the disease. METHODS: Twenty patients with vernal keratoconjunctivitis were randomly chosen from the External Disease and Cornea Sector. They were clinically evaluated, and a score ranging from 0 to 20 was applied based on signs and symptoms on ophthalmic examination. All subjects underwent a skin prick test against standardized allergens, such as house dust mites D. pteronyssinus, D. farinae, and Blomia tropicalis, as well as allergens from cat, dog, fungi and feather. RESULTS: Seventy-five per cent of patients were positive for at least one of the allergens tested. House dust mites were responsible for the majority of the cases (75 percent). There was a poor correlation between the clinical score and sensitivity to allergens (r= - 0.119 for fungi; r= - 0.174 for dog; r= - 0.243 for house dust mites; r= - 0.090 for feather). A significant correlation was found only for cat allergen extract (r = - 0.510; p=0.024). CONCLUSIONS: Our study demonstrated poor correlation between cutaneous hypersensitivity tests and clinical findings in patients with vernal keratoconjuntivitis. We concluded that skin response to inhalant allergens is not a useful test to identify clinical severity and chronicity of inflammatory process in this disease.

OBJETIVO: Avaliar o papel da sensibilização cutânea a diferentes aeroalérgenos em pacientes com ceratoconjuntivite vernal e a correlação entre esta e os aspectos clínicos da doença. MÉTODOS: Vinte pacientes do setor de doenças externas e córnea foram aleatoriamente convidados para participar deste estudo. Os pacientes foram avaliados e a eles foi atribuído um escore clínico variando de 0 a 20 de acordo com sinais e sintomas presentes no exame oftalmológico. Todos os pacientes foram submetidos a testes cutâneos de hipersensibilidade imediata contra aeroalérgenos padronizados como os ácaros domiciliares D. pteronyssinus, D. farinae e Blomia tropicalis, assim como também a alérgenos de epitélio de gato, epitélio de cão, mistura de fungos e mistura de penas. RESULTADOS: Setenta e cinco por cento dos pacientes tiveram teste de hipersensibilidade imediata positivo contra pelo menos um dos antígenos testados. Os ácaros domiciliares foram responsáveis pela maioria destes casos (75 por cento). Houve uma pobre correlação entre o escore clínico e a hipersensibilidade cutânea aos alérgenos (r= -0,119 para fungos; r= -0,174 para epitélio de cão; r= -0,243 para ácaros domiciliares; r= -0,090 para mistura de penas). Houve correlação significativa apenas contra epitélio de gatos (r= -0,510; p=0,024). CONCLUSÃO: O estudo demonstrou uma pobre correlação entre os testes cutâneos de hipersensibilidade imediata e os achados clínicos nos pacientes com ceratoconjuntivite vernal. Os testes cutâneos de hipersensibilidade imediata contra aeroalérgenos não foi parâmetro eficaz na identificação dos casos de maior gravidade e cronicidade de ceratoconjuntivite vernal.