Carnitine analysis in pterygium / Análise de carnitina no pterígio

ABSTRACT Purpose: The aim of the present study was to measure the free carnitine and acylcarnitine levels in pterygium tissue and normal conjunctival tissue at the metabolomics level using tandem mass spectrometry. Methods: In this prospective, clinical randomized study, pterygium tissues and normal conjunctival tissues taken during pterygium excision with autograft were compared regarding their free carnitine and acylcarnitine profiles. After tissue homogenization, carnitine levels were measured using tandem mass spectrometry. The data were statistically analyzed with the Wilcoxon signed-rank test. Results: Pterygium and normal conjunctival tissue samples from a single eye of 29 patients (16 females, 13 males; mean age, 54.75 ± 11.25 years [range, 21-78 years]) were evaluated. While the free carnitine (C0) level was significantly high in the pterygium tissue (p<0.001), acylcarnitine levels were significantly high in some esterized derivatives (C2, C5, C5:1, C5DC, C16:1, C18, methylglutarylcarnitine) (p<0.05). No statistically significant difference was determined for the other esterized derivatives (p>0.05). Conclusion: That the carnitine levels in pterygium tissue were higher suggests that acceleration of cell metabolism developed secondary to chronic inflammation and the premalignant characteristics of pterygium tissue. High carnitine levels may also effectively suppress the apoptosis process. The data reported in our study indicate that further, more extensive studies of the carnitine profile could help clarify the pathogenesis of pterygium.

RESUMO Objetivo: O objetivo deste estudo foi medir os níveis de carnitina livre e acil-carnitina a nível metabolómico com espectrometria de massa em tandem no tecido do pterígio e no tecido conjuntivo normal. Método: Neste estudo prospetivo, clínico e aleatório, os tecidos de pterígio e os tecidos normais de conjuntiva, retirados durante a cirurgia de pterígio com autoenxerto, foram comparados em relação ao perfil de carnitina livre e de acil-carnitina. Após a homogeneização dos tecidos, os níveis de carnitina foram medidos por espectrometria de massa em tandem. A análise estatística dos dados foi realizada com o teste dos postos sinalizados de Wilcoxon. Resultados: A avaliação foi feita através de amostras de tecido pterígio e de conjuntiva normal de um único olho de 29 pacientes (16 mulheres, 13 homens). A média de idade dos pacientes foi de 54,75 ± 11,25 anos (faixa dos 21 aos 78 anos). Enquanto o nível de carnitina livre (C0) foi significativamente elevado no tecido pterígio (p<0,001), os níveis de acil-carnitina foram significativamente elevados em alguns derivados esterificados (C2, C5, C5: 1, C5DC, C16:1, C18, metilglutaril carnitina) (p<0,05). Não foi determinada uma diferença estatisticamen te significante noutros derivados esterificados (p>0,05). Conclusão: Os níveis mais elevados de carnitina no tecido do pterígio sugerem que a aceleração do metabolismo celular se tenha tornado secundária com o efeito da inflamação crónica e o caráter pré-maligno do tecido do pterígio. Os níveis elevados de carnitina também podem ser eficazes na supressão do processo de apoptose. Os dados obtidos no estudo indicam que estudos mais extensivos do perfil da carnitina contribuiriam para o esclarecimento da patogénese do pterígio.

Expression of specificity protein 1 and collagen I in primary pterygial tissues / Expressão da proteína de especificidade 1 e colágeno I em tecidos pterigiais primários

ABSTRACT Purpose: To determine the expression profiles of the transcription factor specificity protein 1 and collagen I in primary pterygial and normal conjunctival tissues, and to explore the role of specificity protein 1 and collagen I in pterygial development. Methods: The pterygial tissues of 20 patients who underwent resection of primary pterygial tissue in our hospital from June 2016 to December 2017 and the conjunctival tissues of 10 patients with enucleation due to trauma were collected. Reverse transcription quantitative-po lymerase chain reaction and western blot analyses were used to detect the relative expression levels of specificity protein 1 and type I collagen at the mRNA and protein levels. Results: The content of specificity protein 1 and collagen I mRNA and protein was significantly greater in primary pterygial tissue than it was in conjunctival tissue (p<0.05). There was a positive correlation between the mRNA and protein levels of specificity protein 1 and collagen I in primary pterygial tissues (protein: r=1, p<0.05; mRNA: r=1, p<0.05). Conclusion: Specificity protein 1 and collagen I are expressed in normal conjunctival and pterygial tissues, but expression is significantly greater in the latter. Specificity protein 1 and collagen I may be involved in the regulation of the development of primary pterygium.

RESUMO Objetivo: Determinar os perfis de expressão do fator de transcrição da proteína de especificidade 1 e do colágeno I em tecidos pterigiais primários e conjuntivais normais, e explorar o papel da proteína de especificidade 1 e colágeno I no desenvolvimento pterigial. Métodos: Foram coletados os tecidos pterigiais de 20 pacientes submetidos à ressecção de tecido de pterígio primário em nosso hospital no período de junho de 2016 a dezembro de 2017 e os tecidos conjuntivais de 10 pacientes com enucleação por trauma. A reação em cadeia da polimerase quantitativa de transcriptase reversa e a análise de Western blot foram utilizadas para detectar os níveis de expressão relativa da proteína de especificidade 1 e colágeno tipo I nos níveis de mRNA e proteína. Resultados: O conteúdo de especificidade da proteína 1 e do mRNA e proteína do colágeno I foi significativamente maior no tecido de pterígio primário do que no tecido conjuntival (p<0,05). Houve correlação positiva entre os níveis de mRNAs e proteína de especificidade 1 e colágeno I nos tecidos primários do pterígio (proteínas: r=1, p<0,05; mRNA: r=1, p<0,05). Conclusão: A proteína de especificidade 1 e do colágeno I é expressa nos tecidos conjuntivais e pterigiais normais, mas a expressão é significativamente maior no segundo. A especificidade da proteína 1 e do colágeno I pode ser envolvida na regulação do desenvolvimento do pterígio primário.

Preoperative tranilast as adjunctive therapy to primary pterygium surgery with a 1-year follow-up / Terapia auxiliar com o tranilast pré-operatório na cirurgia do pterígio primário com um ano de seguimento

Purpose: To determine the efficacy of tranilast as an adjunctive therapy in conjunctival autograft. Methods: Twenty-nine patients were randomly allocated to the Tranilast Group (n=15) or the Control Group (n=14). The Tranilast Group received a subconjunctival injection of 0.5% tranilast 30 days prior to surgery. Conjunctival autograft was performed in both groups using fibrin sealant and 0.02% subconjunctival mitomycin C at the end of the surgery. After the resection of the pterygium, immunohistochemistry was performed with 100 cells to identify epithelial cells positive for transforming growth factor-β (TGF-β). Subjective symptoms were evaluated using a 5-point scale, and the recurrence rate was assessed. Results: Both groups showed improvements in their symptoms and similar clinical results. Compared with the Control Group, the Tranilast Group failed to show a decreased recurrence rate (p=0.59). However, the number of epithelial cells expressing TGF-β was lower in the Tranilast Group (5 cells; 95% CI: 2.56-13.15; Control Group, 16 cells, 95% CI: 11.53-24.76; p=0.01). Minimal but reversible complications, including glaucoma secondary to corticosteroids and granuloma, occurred during the study. Conclusion: Tranilast was effective in decreasing the number of pterygium epithelial cells expressing TGF-β. .

Objetivo: Determinar a eficácia do tranilast, como terapia auxiliar no transplante autólogo de conjuntiva. Métodos: Vinte e nove pacientes foram randomizados em dois grupos: Grupo Tratado (15) e Grupo Controle (14). Trinta dias antes da cirurgia, o Grupo Tratado recebeu uma injeção subconjuntival de tranilast a 0,5%. O transplante autólogo de conjuntiva foi realizado em ambos os grupos, usando-se a cola de fibrina e a mitomicina 0,02% subconjuntival, ao final da cirurgia. Cada paciente foi examinado por 12 meses de acompanhamento. A imuno-histoquímica foi realizada, mediante um total de 100 células, a fim de que se contassem as células epiteliais positivas, para o fator de crescimento transformador beta (TGF-β), após a cirurgia do pterígio. Os sintomas subjetivos foram avaliados usando-se uma escala de cinco pontos, e a taxa de recorrência foi avaliada. Resultados: Os 2 grupos apresentaram melhora dos sintomas e com resultados clínicos similares. Quando comparado com o Grupo Controle, o Grupo Tratado falhou em mostrar uma diminuição da taxa de recorrência (p=0,59). Entretanto o número de células epiteliais expressando o TGF-β foi menor no Grupo Tratado (5 células; 95% CI=2,56-13,15; Grupo Controle, 16 células; 95% CI: 11,53-24,76, p=0,01). Complicações mínimas, mas reversíveis, ocorreram durante o estudo, incluindo glaucoma secundário ao uso de corticoide e granuloma. Conclusão: O tranilast foi efetivo em diminuir o número células epiteliais do pterígio expressando o TGF-β. .

Transconjunctival penetration of mitomycin C.

AIMS: The study was performed to estimate transconjunctival penetration of mitomycin C (MMC) to Tenon's tissue following application over the intact conjunctiva before routine trabeculectomy. SETTINGS AND DESIGN: Institution-based case series. MATERIALS AND METHODS: In 41 eyes of 41 patients, MMC (0.4 mg/ml for 3 min) was applied over the intact conjunctiva before beginning trabeculectomy. Tenon's capsule directly beneath the site of application was excised during trabeculectomy and was homogenized, centrifuged and MMC concentrations were analyzed using high-performance liquid chromatography (HPLC). Statistical Analysis Used: Statistical analysis was performed using STATA 8.0 version software (STATA Corporation, Houston, TX, USA). In this study, P -values less than 0.05 were considered as statistically significant. RESULTS: The average weight of the sample of Tenon's tissue excised was 5.51+/-4.42 mg (range: 0.9-17.1) and the average estimated MMC concentration found to be present in Tenon's tissue using HPLC was 18.67+/-32.36 x 10(-6) moles/kg of the tissue (range: 0.38-197.05 x 10(-6)). In 36 of the 41 patients (87.80%), the MMC concentration reached above 2 x 10(-6) moles/kg of the tissue concentration required to inhibit human conjunctival fibroblasts. CONCLUSIONS: Mitomycin C does permeate into the subconjunctival tissue after supraconjunctival application for 3 min. Application of MMC over the conjunctiva may be a useful alternative to subconjunctival or subscleral application during routine trabeculectomy and as an adjunct for failing blebs.

Effects of Smoking on Tear Film and Ocular Surface

This study was performed to evaluate the changes of tear film and ocular surface caused by smoking. Symptom scoring, tear film break-up time (BUT), basal tear secretion test, corneal sensitivity test, keratoepitheliopathy scoring, and conjunctival impression cytology were performed in 29 smokers (58 eyes) and 26 non-smokers (52 eyes). Tear film BUT, basal tear secretion, corneal sensitivity, and squamous metaplasia were 7.71 +/- 2.66 sec, 6.29 +/- 2.85 mm, 53.69 +/- 5.69 mm, and 2.45 +/- 1.26 in smokers and 9.62 +/- 3.14 sec, 10.04 +/- 3.87 mm, 56.46 +/- 4.79 mm, and 1.12 +/- 0.83 in non-smokers, respectively (p< 0.05). Symptom score, keratoepitheliopathy score, and goblet cell density were not significantly different between the two groups. Tear film BUT was shorter, basal tear secretion and corneal sensitivity were lower, and squamous metaplasia was higher in heavy smokers than in light smokers. In conclusion, smoking deteriorates the tear film and ocular surface with decreased quantity and quality of tear film, decreased corneal sensitivity, and squamous metaplasia, and this deterioration is related to the amount of smoking.

Paracoccidioidomicose primária de pálpebra e conjuntiva / Primary paracoccidioidomycosis of eyelid and conjunctiva

A paracoccidioidomicose (Pbmicose) é a micose profunda mais frequente no Brasil, mas o acometimento ocular é raro e, quando presente, geralmente secundário. Os autores relatam caso de Pbmicose pálpebro-conjuntival em que näo foi encontrado foco extra-ocular. É feita uma revisäo da literatura e discutida a importância da suspeita diagnóstica em populaçäo de risco e do tratamento precoce desta condiçäo para o bom prognóstico visual.

Increased numbers of Langerhans cell and expression of HLA-Dr antigen in the giant papilla of patients with giant papillary conjunctivitis

A study of histopathologic changes, ultrastructure, and expression of the HLA-Dr antigen within the giant papillae of patients with giant papillary conjunctivitis was performed to determine whether cell-mediated immune response is related to this condition. Conjunctival giant papillae from ten patients with giant papillary conjunctivitis were examined by light and electron microscopy and by the indirect immunofluorescent staining method with HLA-Dr antibody. The infiltration of eosinophilic neutrophils and granules was most prominent, with the occasional infiltration of mast cells, as shown by light microscopy. The infiltration of activated fibroblasts and Langerhans cells was also observed. Cells expressing HLA-Dr antigen were also markedly increased, as shown by the immunofluorescent method. These findings suggest that delayed hypersensitivity may, along with the processes of antigen presentation by HLA-Dr-expressing (including Langerhans) cells, contribute to the pathogenesis of giant papillary conjunctivitis.

Eosinofilia en enfermos de uveitis anterior aguda / Eosinophilia in patients presenting with acute anterior uveitis

Se realizo un estudio prospectivo en 50 enfermos de uveitis anterior aguda con el proposito de describir en los mismos la eosinofilia ocular y en sangre. En el estudio se corrobora que en la totalidad de los casos con sensibilidad alergica demostrada, aparece eosinofilia ocular y en sangre. Sin embargo, en aquellos en que la etiologia fue una colecistopatia con giardiasis cronica asociada, aparece eosinofilia ocular en el 79,0 por ciento. Es destacable que en los pacientes cuya causa fue un poco septico,toxoplasmosis o enfermedad autoinmune, no se contato eosinofilia. En dicha investigacion se valora la importancia del eosinofilo en los pacientes con uveitis anterior aguda, cuya causa probable es alergica, infecciosa o autoinmunitaria