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.

COVID-19 Ocular Manifestations in the Early Phase of Disease.

Purpose: To evaluate and establishe the number of patients with ocular manifestations in the early phase of systemic COVID-19 infection.Methods: A cross-sectional study was performed in a COVID-19 referral center regarding ocular findings in patients with COVID-19 in the first few days of the disease. The patients were submitted to a clinical examination, an ophthalmic exam and a RT-PCR for SARS-COV-2.Results: Out of 1740 patients, we reported 108 patients with ocular manifestations. Forty-nine with markedly conjunctivitis had conjunctival swab positive for SARS-COV-2, four of them developed keratitis. There were mostly no evidence of retinopathy nor decrease in visual acuity. They had no marked clinical symptoms, which can contribute and demonstrate that the virus may cause ocular disease as an only finding or in the very early stage of the infection.Conclusion: Patients were in the first days of COVID-19 infection, presented ocular manifestations suggested to be related to the virus and need to be aware of the pathways of transmissions.

Conjunctivitis Related to not Severe COVID-19: A Case Report.

PURPOSE: The purpose of this article is to report a bilateral conjunctivitis case in a patient with confirmed 2019 novel coronavirus disease (COVID-19). CASE REPORT/OBSERVATIONS: A 24-year-old healthy man developed severe conjunctivitis in his right eye, followed by his left eye 4 days after fever and myalgia. He reported yellowish conjunctival discharge, foreign body sensation, redness, and tearing, and those symptoms were resolved in 10 days. Analyses of nasopharyngeal secretions identified SARS-CoV-2 by polymerase chain reaction (PCR). CONCLUSION: Mild COVID-19 can cause severe bilateral conjunctivitis. The ophthalmologist may be aware of conjunctival secretions, a possible path of infection, during eye checkup.

Relations between the ocular surface and SARS-CoV-2 / Relações entre superfície ocular e SARS-CoV-2

ABSTRACT Introduction: The SARS-CoV-2 pandemic has been a major challenge for the international scientific community. Since its inception, studies aiming to describe pathophysiological aspects and clinical manifestations of the disease have been conducted, raising hypotheses and confirming possible associations. One aspect of this scientific medical production is the role of the ocular surface as a means of transmission and clinical presentation of viral syndrome. Objectives: To analyze the role of the ocular surface in transmission, pathophysiology, and clinical manifestations of SARS-CoV-2, by means of a systematic review. Methods: The search was carried out in three databases: Cochrane, PubMed Central Journals and MEDLINE, using the following descriptors: "COVID-19, ophthalmology". The filters last five years and studies on humans resulted in 32 studies; in that 12 were excluded for not meeting the purpose of the study. Results: There are still few published studies on the relation between SARS-CoV-2 and the ocular route. Most studies showed an association between the presence of nonspecific ocular manifestations and infection by the new coronavirus, with limitations in the number of patients analyzed and the methodology adopted. Hypotheses about the pathophysiological role are largely anchored in the association of SARS-CoV and the ocular surface evaluated in the past. Comments: The results found are still not sufficient to confirm the role of the ocular surface in the pathophysiology of the disease. Most of these preliminary studies are of considerable importance in raising hypotheses based on the medical analysis of the patients studied. However, larger studies with standardized methodology for diagnostic protocol and laboratory analysis of the individuals assessed are required.

RESUMO Introdução: A pandemia da SARS-CoV-2 tem sido um grande desafio para a comunidade científica internacional. Desde seu surgimento, estudos com a intenção de descrever os aspectos fisiopatológicos e as manifestações clínicas da doença vêm sendo conduzidos, levantando hipóteses e confirmando possíveis associações. Um dos temas dessa produção médica científica é o papel da superfície ocular como meio de transmissão e apresentação clínica da síndrome viral. Objetivo: Analisar o papel da superfície ocular na transmissão, na fisiopatologia e nas manifestações clínicas de SARS-CoV-2, através de uma revisão sistemática. Realizou-se a busca em três bancos de dados Cochrane Database, PubMed® e MEDLINE®, utilizando os descritores "COVID-19 e ophthalmology". Foram definidos como filtros o artigo ter sido publicado nos últimos 5 anos e estudo realizado em humanos, tendo sido encontrados 32 artigos. Destes, foram excluídos 12 por não corresponderem ao objetivo do estudo. Resultados: Ainda são poucos os estudos publicados sobre a relação entre o coronavírus 2 da síndrome respiratória aguda grave (SARS-CoV-2) e a via ocular. A maioria dos estudos mostrou associação entre a presença de manifestações oculares inespecíficas e a infecção pelo novo coronavírus, com limitações no número de pacientes analisados e na metodologia adotada. Hipóteses sobre o papel fisiopatológico se ancoram, em grande parte, na associação estudada entre o SARS-CoV-2 e a superfície ocular no passado. Comentários: Os resultados encontrados ainda não são suficientes para confirmar o papel da superfície ocular na fisiopatologia da doença. Grande parte desses estudos preliminares têm importância considerável ao levantar hipóteses baseadas na análise clínica dos pacientes estudados. No entanto, são necessários estudos maiores e com metodologia padronizada para protocolo diagnóstico e análise laboratorial dos indivíduos avaliados.

COVID-19 and the eye: how much do we really know? A best evidence review.

To identify and classify available information regarding COVID-19 and eye care according to the level of evidence, within four main topics of interest: evidence of the virus in tears and the ocular surface, infection via the conjunctival route, ocular manifestations, and best practice recommendations. A structured review was conducted in PubMed, ScienceDirect, LILACS, SciELO, the Cochrane Library and Google Scholar on COVID-19 and ophthalmology. The Oxford Centre for Evidence Based Medicine 2011 Levels of Evidence worksheet was used for quality assessments. 1018 items were identified in the search; 26 records were included in the qualitative synthesis, which encompassed 6 literature reviews, 10 case series or cross-sectional studies, 4 case reports, and 6 intervention descriptions. Seventeen out of 26 records (65%) were categorized as level 5 within the Oxford CBME methodology grading system, the rest were level 4. The evidence generated on COVID-19 and ophthalmology to date is limited, although this is understandable given the circumstances. Both the possible presence of viral particles in tears and conjunctiva, and the potential for conjunctival transmission remain controversial. Ocular manifestations are not frequent and could resemble viral infection of the ocular surface. Most recommendations are based on the strategies implemented by Asian countries during previous coronavirus outbreaks. There is a need for substantive studies evaluating these strategies in the setting of SARS-CoV-2. In the meantime, plans for applying these measures must be implemented with caution, taking into account the context of each individual country, and undergo regular evaluation.

COVID-19 and the eye: how much do we really know? A best evidence review / COVID-19 e o olho: quanto sabemos realmente? Uma revisão das melhores evidências

ABSTRACT To identify and classify available information regarding COVID-19 and eye care according to the level of evidence, within four main topics of interest: evidence of the virus in tears and the ocular surface, infection via the conjunctival route, ocular manifestations, and best practice recommendations. A structured review was conducted in PubMed, ScienceDirect, LILACS, SciELO, the Cochrane Library and Google Scholar on COVID-19 and ophthalmology. The Oxford Centre for Evidence Based Medicine 2011 Levels of Evidence worksheet was used for quality assessments. 1018 items were identified in the search; 26 records were included in the qualitative synthesis, which encompassed 6 literature reviews, 10 case series or cross-sectional studies, 4 case reports, and 6 intervention descriptions. Seventeen out of 26 records (65%) were categorized as level 5 within the Oxford CBME methodology grading system, the rest were level 4. The evidence generated on COVID-19 and ophthalmology to date is limited, although this is understandable given the circumstances. Both the possible presence of viral particles in tears and conjunctiva, and the potential for conjunctival transmission remain controversial. Ocular manifestations are not frequent and could resemble viral infection of the ocular surface. Most recommendations are based on the strategies implemented by Asian countries during previous coronavirus outbreaks. There is a need for substantive studies evaluating these strategies in the setting of SARS-CoV-2. In the meantime, plans for applying these measures must be implemented with caution, taking into account the context of each individual country, and undergo regular evaluation.

RESUMO Identificar e classificar as informações disponíveis sobre o COVID-19 e o tratamento oftalmológico de acordo com o nível de evidência, dentro de quatro tópicos principais de interesse: evidência do vírus nas lágrimas e na superfície ocular, infecção pela via conjuntival, manifestações oculares e recomendações de melhores práticas. Foi realizada uma revisão estruturada no PubMed, ScienceDirect, LILACS, SciELO, Biblioteca Cochrane e Google Scholar no COVID-19 e oftalmologia. A planilha de Níveis de Evidência 2011 do Oxford Centre for Evidence Based Medicine 2011 foi usada para avaliações de qualidade. Mil e dezoito itens foram identificados na busca; Foram incluídos 26 registros na síntese qualitativa, que incluiu 6 revisões de literatura, 10 séries de casos ou estudos transversais, 4 relatos de casos e 6 descrições de intervenções. Dezessete dos 26 registros (65%) foram classificados como nível 5 no sistema de classificação da metodologia Oxford CBME, o restante foi no nível 4. As evidências geradas no COVID-19 e na oftalmologia até o momento são limitadas, embora isso seja compreensível dadas as circunstâncias. Tanto a possível presença de partículas virais em lágrimas e conjuntiva quanto o potencial de transmissão conjuntival permanecem controversos. As manifestações oculares não são frequentes e podem se assemelhar a infecção viral da superfície ocular. A maioria das recomendações baseia-se nas estratégias implementadas pelos países asiáticos durante surtos anteriores de coronavírus. Há necessidade de estudos aprofundados avaliando essas estratégias no cenário da SARS-CoV-2. Enquanto isso, os planos para a aplicação dessas medidas devem ser implementados com cautela, levando em consideração o contexto de cada país e submetidos a auditorias periódicas.

Molecular Detection of Feline Leukemia Virus in Oral, Conjunctival, and Rectal Mucosae Provides Results Comparable to Detection in Blood.

Feline leukemia virus (FeLV) infection causes immunosuppression, degeneration of the hematopoietic system, and fatal neoplasms. FeLV transmission occurs mainly by close social contact of infected and susceptible cats. Developing procedures for the diagnosis of feline retroviruses is crucial to reduce negative impacts on cat health and increase the number of animals tested. Blood collection requires physical or chemical restraint and is usually a stressful procedure for cats. Our objective was to evaluate the use of samples obtained from oral, conjunctival, and rectal mucosae for the molecular diagnosis of FeLV. Whole blood and oral, conjunctival, and rectal swabs were collected from a total of 145 cats. All samples were subjected to the amplification of a fragment of the gag gene of proviral DNA. Compared to blood samples used in this study as a reference, the accuracies for each PCR were 91.72, 91.23, and 85.50% for samples obtained by oral, conjunctival, and rectal swabs, respectively. The diagnostic sensitivity and specificity were 86.11 and 97.26% for the oral swabs, 90 and 92.59% for the conjunctival swabs, and 74.24 and 95.77% for the rectal swabs, respectively. The kappa values for oral, conjunctival, and rectal swabs were 0.834, 0.824, and 0.705, respectively. The diagnosis of these samples showed the presence of proviral DNA of FeLV in oral and conjunctival mucosae. In conclusion, mucosal samples for the molecular diagnosis of FeLV are an excellent alternative to venipuncture and can be safely used. It is faster, less laborious, less expensive, and well received by the animal.

AmpliVue Is a Practical and Timely Test for the Detection of HSV From Keratitis Specimens.

OBJECTIVE: The laboratory diagnostic detection of herpes simplex virus (HSV) from eye samples must be practical, timely, and definitive for appropriate therapy. Although polymerase chain reaction (PCR) and/or cell culture can be definitive, HSV results can be delayed. Enzyme Linked Virus Inducible System (ELVIS) is a test that can provide results within 24 to 48 hr. We evaluated "AmpliVue HSV 1+2 Assay" as a molecular colorimetric test that can detect HSV (1 or 2) DNA within 1 hr. METHODS: Cornea/conjunctival samples were tested retrospectively with AmpliVue against 53 true-positive and 20 true-negative specimens collected in chlamydial transport medium. All clinical specimens were tested by cell culture isolation, PCR, and ELVIS for routine patient care. RESULTS: The sensitivity of AmpliVue against ocular samples that were both culture-positive and PCR-positive was 84%. The specificity of AmpliVue was 100%. Only one clinical sample was HSV-2 positive, whereas all others tested positive for HSV-1. Based on PCR-positive and cell culture-negative samples, AmpliVue (11 of 17) tested more positive than ELVIS (0 of 17) (P=0.003, Fisher Exact). CONCLUSIONS: AmpliVue is moderately sensitive and highly specific as a practical and timely diagnostic test for detecting ocular HSV. Expertise is readily achieved and the test is straightforward with easy interpretation. Negative AmpliVue testing must be confirmed with PCR. AmpliVue has potential as an office-based diagnostic test.

Molecular epidemiology of adenovirus conjunctivitis in Rio de Janeiro, Brazil, between 2004 and 2007.

Viral conjunctivitis is a common, highly contagious disease often caused by adenovirus. We investigate the frequency of adenoviral conjunctivitis in the population of Rio de Janeiro, Brazil, between March 2004 and May 2007 and identified the predominant serotype circulating among this population. Seventy-five ocular swabs were collected from 66 patients with clinical presentation of conjunctivitis. The specimens were analyzed for detection of adenovirus (AdV) by polymerase chain reaction (PCR). The PCR products were further analyzed for virus typing by sequence analysis and/or heteroduplex mobility assay (HMA). Forty-five samples (60%) were positive for AdV of which 21 samples were typed as AdV19 (46.7%), 7 AdV8 (15.5%), 3 AdV31 (6.7%), and one each AdV1, AdV2, AdV3, AdV4 and AdV6. For nine samples the serotype was not determined. AdV19 was the predominant serotype circulating in Rio de Janeiro during the studied period.

Molecular epidemiology of adenovirus conjunctivitis in Rio de Janeiro, Brazil, between 2004 and 2007 / Epidemiologia molecular de adenovírus associados à conjuntivite no Rio de Janeiro, Brasil, no periodo de 2004 a 2007

Viral conjunctivitis is a common, highly contagious disease often caused by adenovirus. We investigate the frequency of adenoviral conjunctivitis in the population of Rio de Janeiro, Brazil, between March 2004 and May 2007 and identified the predominant serotype circulating among this population. Seventy-five ocular swabs were collected from 66 patients with clinical presentation of conjunctivitis. The specimens were analyzed for detection of adenovirus (AdV) by polymerase chain reaction (PCR). The PCR products were further analyzed for virus typing by sequence analysis and/or heteroduplex mobility assay (HMA). Forty-five samples (60%) were positive for AdV of which 21 samples were typed as AdV19 (46.7%), 7 AdV8 (15.5%), 3 AdV31 (6.7%), and one each AdV1, AdV2, AdV3, AdV4 and AdV6. For nine samples the serotype was not determined. AdV19 was the predominant serotype circulating in Rio de Janeiro during the studied period.

A conjuntivite viral é doença ocular comum, altamente contagiosa, frequentemente causada por adenovírus. Neste estudo, investigamos a frequência de conjuntivite por adenovírus na população do Rio de Janeiro, Brasil, entre março de 2004 e maio de 2007, e identificamos o sorotipo predominante circulando nesta população. Setenta e cinco swabs de secreção ocular foram coletados de 66 pacientes com conjuntivite. As amostras foram analisadas para detecção de adenovírus (AdV) por reação em cadeia da polimerase (PCR). Os produtos da PCR foram caracterizados por sequenciamento e/ou ensaio de mobilidade do heteroduplex (Heteroduplex Mobility Assay - HMA) para identificação do sorotipo viral. Quarenta e cinco (60%) amostras foram positivas para AdV das quais 21 foram identificadas como pertencentes ao sorotipo AdV19 (46,7%), sete AdV8 (15,5%), três AdV31 (6,7%), e uma de cada: AdV1, AdV2, AdV3, AdV4 e AdV6. Para nove amostras o sorotipo não pode ser identificado. O AdV 19 foi o sorotipo predominante circulando no Rio de Janeiro durante o período estudado.

[Use conditions of boric acid solution in the eye: handling and occurrence of contamination]. / Uso ocular de água boricada: condições de manuseio e ocorrência de contaminação.

PURPOSE: To evaluate use conditions and detect contamination in bottles of boric acid solution. METHODS: A convenience sample of 42 recruited patients using boric acid solution came to the Ophthalmology Emergency Room of the São Paulo Hospital from February to March of 2003. Cultures were taken from material of the conjunctival sac, inner surface of bottle edge, inner part of cap and from 1 ml of boric acid solution of each bottle. RESULTS: Of the 42 boric acid solution bottles, 17 (40.5%) showed contamination: 1 (2.4%) in the solution, 17 (40.5%) in the inner cap and 6 (14.3%) in the inner part of the bottle edge. Of the 17 contaminated bottles, 10 (58.8%) were handled inappropriately and 13 (76.5%) of the bottles were not discharged after first use. The most common microorganisms found in the caps and edges of the bottles were Staphylococcus sp (69.6%), followed by Gram-positive bacillus (26.1%). Sixteen bottles (38.1%) had been opened more than a month ago and 5 (31.3%) of those showed contamination. The boric acid solution bottle directions shown on the labels were incomplete and not clear. The use of boric acid solution was on recommendation of their own, friends or relatives in 26 (61.9%) cases; pharmacists in 8 (19.0%) cases, ophthalmologists in 5 (11.9%) cases and general practitioners in 3 (7.1%) cases. CONCLUSION: In most cases, the topic use of boric acid solution was recommended by non-physicians. The bottles, in general, were handled inappropriately, and hence presented a much higher level of contamination that did the boric acid solution inside. The lower level of contamination in the solution is possibly associated with the anti-septic characteristics of the boric acid solution.

Rapid diagnosis of acute hemorrhagic conjunctivitis due to coxsackievirus A24 variant by real-time one-step RT-PCR.

Coxsackievirus A24 variant is, together with enterovirus 70 and adenoviruses, the major etiological agent involved in acute hemorrhagic conjunctivitis outbreaks worldwide. However, the standard virus isolation method followed by serotyping or VP1 region sequencing is time-consuming. A rapid method for the detection of coxsackievirus A24 variant from conjunctival swab specimens would be useful in the context of explosive and extensive outbreaks. A one-step real-time RT-PCR assay based on TaqMan technology was thus developed and assessed on 36 conjunctival swabs from outbreaks of conjunctivitis in Morocco in 2004 due to a coxsackievirus A24 variant and in Corsica in 2006 due to adenovirus type 3, and 83 virus strains including 41 coxsackievirus A24 variant collected in French Guiana and Guadeloupe in 2003, in the Democratic Republic of the Congo in 2003, in Morocco in 2004 and 42 other virus species genetically close or known to be responsible for conjunctivitis. All the conjunctival swabs from coxsackievirus A24 variant related outbreak and the 41 coxsackievirus A24 variant strains were tested positive by the RT-PCR assay within 4h. This novel single-tube real-time RT-PCR assay is sensitive and specific, and consists in a reliable and faster alternative to the viral culture for recent and future acute hemorrhagic conjunctivitis outbreaks caused by coxsackievirus A24 variant.

Uso ocular de água boricada: condições de manuseio e ocorrência de contaminação / Use conditions of boric acid solution in the eye: handling and occurrence of contamination

OBJETIVOS: Avaliar as condições de uso de água boricada e verificar a contaminação dos frascos e seu conteúdo. MÉTODOS: Foram selecionados, por critério de conveniência, quarenta e dois pacientes, usuários de água boricada, que compareceram ao Pronto-Socorro de Oftalmologia do Hospital São Paulo, em fevereiro e março de 2003. Foi colhido material para cultura do saco conjuntival, da superfície interna da borda do frasco, da superfície interna da tampa, além de 1 ml de solução do frasco. RESULTADOS: Dos 42 recipientes de água boricada, 17 (40,5 por cento) apresentavam contaminação, sendo 1 (2,4 por cento) no conteúdo liquido, 17 (40,5 por cento) na parte interna da tampa e 6 (14,3 por cento) na parte interna da borda do frasco. Dos 17 frascos contaminados, 10 (58,8 por cento) tiveram suas tampas manuseadas de maneira inadequada e 13 (76,5 por cento) frascos já haviam sido usados em outras ocasiões. Os microrganismos mais encontrados nas tampas e bordas foram Staphylococcus sp (69,6 por cento) e bacilos Gram-positivos (26,1 por cento). Dezesseis (38,1 por cento) frascos foram abertos há mais de um mês e, destes, 5 (31,3 por cento) apresentaram contaminação. A instrução de uso nos rótulos dos frascos era inconsistente. A utilização de água boricada foi por conta própria, por indicação de amigos ou parentes em 26 (61,9 por cento) casos; indicação de farmacêuticos em 8 (19,0 por cento); de oftalmologistas em 5 (11,9 por cento) e de clínicos gerais em 3 (7,1 por cento). CONCLUSÃO: A indicação de uso tópico oftálmico de água boricada foi feita, na maioria, por leigos. Os frascos, em geral, eram manipulados de maneira inadequada, apresentando contaminação em uma proporção de casos muito maior do que a contaminação do líquido. Essa porcentagem menor de contaminação do conteúdo provavelmente está associada às características anti-sépticas do produto.

PURPOSE: To evaluate use conditions and detect contamination in bottles of boric acid solution. METHODS: A convenience sample of 42 recruited patients using boric acid solution came to the Ophthalmology Emergency Room of the São Paulo Hospital from February to March of 2003. Cultures were taken from material of the conjunctival sac, inner surface of bottle edge, inner part of cap and from 1 ml of boric acid solution of each bottle. RESULTS: Of the 42 boric acid solution bottles, 17 (40.5 percent) showed contamination: 1 (2.4 percent) in the solution, 17 (40.5 percent) in the inner cap and 6 (14.3 percent) in the inner part of the bottle edge. Of the 17 contaminated bottles, 10 (58.8 percent) were handled inappropriately and 13 (76.5 percent) of the bottles were not discharged after first use. The most common microorganisms found in the caps and edges of the bottles were Staphylococcus sp (69.6 percent), followed by Gram-positive bacillus (26.1 percent). Sixteen bottles (38.1 percent) had been opened more than a month ago and 5 (31.3 percent) of those showed contamination. The boric acid solution bottle directions shown on the labels were incomplete and not clear. The use of boric acid solution was on recommendation of their own, friends or relatives in 26 (61.9 percent) cases; pharmacists in 8 (19.0 percent) cases, ophthalmologists in 5 (11.9 percent) cases and general practitioners in 3 (7.1 percent) cases. CONCLUSION: In most cases, the topic use of boric acid solution was recommended by non-physicians. The bottles, in general, were handled inappropriately, and hence presented a much higher level of contamination that did the boric acid solution inside. The lower level of contamination in the solution is possibly associated with the anti-septic characteristics of the boric acid solution.

1-Cinnamoyl-3,11-dihydroxymeliacarpin is a natural bioactive compound with antiviral and nuclear factor-kappaB modulating properties.

We have reported the isolation of the tetranortriterpenoid 1-cinnamoyl-3,11-dihydroxymeliacarpin (CDM) from partially purified leaf extracts of Melia azedarach L. (MA) that reduced both, vesicular stomatitis virus (VSV) and Herpes simplex virus type 1 (HSV-1) multiplication. CDM blocks VSV entry and the intracellular transport of VSV-G protein, confining it to the Golgi apparatus, by pre- or post-treatment, respectively. Here, we report that HSV-1 glycoproteins were also confined to the Golgi apparatus independently of the nature of the host cell. Considering that MA could be acting as an immunomodulator preventing the development of herpetic stromal keratitis in mice, we also examined an eventual effect of CDM on NF-kappaB signaling pathway. CDM is able to impede NF-kappaB activation in HSV-1-infected conjunctival cells and leads to the accumulation of p65 NF-kappaB subunit in the cytoplasm of uninfected treated Vero cells. In conclusion, CDM is a pleiotropic agent that not only inhibits the multiplication of DNA and RNA viruses by the same mechanism of action but also modulates the NF-kappaB signaling pathway.

Detection of human papillomavirus in epithelial lesions of the conjunctiva.

CONTEXT: Many factors like exposure to UV radiation, climatic conditions, genetic predisposition, immunological state and, more recently, the presence of HPV have been implicated in the genesis of some lesions of the conjunctiva, especially the carcinoma. OBJECTIVE: To evaluate the presence of HPV DNA in acquired lesions of the conjunctiva and in normal mucosa. TYPE OF STUDY: Cross-sectional study. SETTING: A public university referral center (the Ophthalmology Service of the A.C. Camargo Hospital - A. Prudente Foundation, São Paulo). PARTICIPANTS: Thirty patients with acquired lesions of the conjunctiva and 60 matched controls (by age and sex) were evaluated in this study, from June 1993 to March 1995. PROCEDURES: The detection of HPV DNA in the normal conjunctiva and in acquired lesions was done by the PCR technique and dot blot hybridization. The material was collected by scraping the normal mucosa and the surface of the lesions. A fragment of fresh frozen tissue and paraffin embedded specimens of each lesion were also included. MAIN MEASUREMENTS: The association between the HPV infection and the presence or absence of conjunctival lesions. RESULTS: Sequences of HPV DNA were detected in 4 of the 31 lesions evaluated (12.9%) and in the healthy mucosa of one individual of the control group (1.6%). HPV type 16 was detected in 2 carcinomas and in the normal mucosa of one individual of the control group. HPV type 11 was demonstrated in 2 papillomas of one patient with lesions in both eyes. CONCLUSIONS: The low frequency of HPV DNA found in the lesions of this sample and the detection of the viral genome in the normal mucosa indicate that there is a weak possibility of association between HPV infection and the carcinoma of the conjunctiva.

Detection of human papillomavirus in epithelial lesions of the conjunctiva

CONTEXT: Many factors like exposure to UV radiation, climatic conditions, genetic predisposition, immunological state and, more recently, the presence of HPV have been implicated in the genesis of some lesions of the conjunctiva, especially the carcinoma. OBJECTIVE: To evaluate the presence of HPV DNA in acquired lesions of the conjunctiva and in normal mucosa. TYPE OF STUDY: Cross-sectional study. SETTING: A public university referral center (the Ophthalmology Service of the A.C. Camargo Hospital - A. Prudente Foundation, Sao Paulo). PARTICIPANTS: Thirty patients with acquired lesions of the conjunctiva and 60 matched controls (by age and sex) were evaluated in this study, from June 1993 to March 1995. PROCEDURES: The detection of HPV DNA in the normal conjunctiva and in acquired lesions was done by the PCR technique and dot blot hybridization. The material was collected by scraping the normal mucosa and the surface of the lesions. A fragment of fresh frozen tissue and paraffin embedded specimens of each lesion were also included. MAIN MEASUREMENTS: The association between the HPV infection and the presence or absence of conjunctival lesions. RESULTS: Sequences of HPV DNA were detected in 4 of the 31 lesions evaluated (12.9 percent) and in the healthy mucosa of one individual of the control group (1.6 percent). HPV type 16 was detected in 2 carcinomas and in the normal mucosa of one individual of the control group. HPV type 11 was demonstrated in 2 papillomas of one patient with lesions in both eyes. CONCLUSIONS: The low frequency of HPV DNA found in the lesions of this sample and the detection of the viral genome in the normal mucosa indicate that there is a weak possibility of association between HPV infection and the carcinoma of the conjunctiva

Field investigation of Mycoplasma gallisepticum infections in house finch (Carpodacus mexicanus) eggs and nestlings.

We conducted a field study to investigate the occurrence of Mycoplasma gallisepticum (MG) in eggs and nestlings from nests of house finches (Carpodacus mexicanus). Forty-three nests were located between the months of April and August 1998 and were followed with one to three sampling efforts. Vitelline membrane of fresh eggs, whole embryos, or swabs from the choanal cleft or conjunctiva of nestlings were inoculated into mycoplasma broth for MG isolation and polymerase chain reaction (PCR) testing. No isolation of MG was made from 39 eggs or 110 nestlings sampled during the study. Pooled choanal and conjunctival swab samples from two broods of nestlings, however, tested positive for MG by PCR. None of the nestlings examined showed clinical signs of conjunctivitis, and no nestling mortality could be linked to MG infection. Serologic tests from 37 older nestlings were negative for antibodies to MG. The results suggest transmission of MG is occurring between breeding adults and their dependent offspring (pseudovertical transmission). Evidence supporting transovarian transmission of MG was not found in these house finches.