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A ocorrência de ectasia corneana após ceratectomia fotorrefrativa é uma complicação rara, porém grave, em cirurgia refrativa. Os possíveis fatores de risco não são bem avaliados, mas a opinião atual é que a falha na detecção de ceratocone pré-operatório possa ser o principal motivo. Neste relato, descrevemos um caso de ectasia corneana após ceratectomia fotorrefrativa em paciente apresentando padrão tomográfico suspeito no pré-operatório, mas sem alterações degenerativas associadas a ceratocone patológico, conforme revelado por microscopia confocal in vivo da córnea. Além disso, revisamos, na literatura, relatos de casos elegíveis de ectasia pós-ceratectomia fotorrefrativa para encontrar características semelhantes. [ABSTRACT FROM AUTHOR]

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Background Diabetic neuropathy (DN) is a very common clinical condition throughout the world. The diagnostic tests currently recommended have low sensitivity, such as electromyography, or are invasive, such as skin biopsy. New techniques have been developed to identify the early involvement of the peripheral nerve. With the advent of corneal confocal microscopy (CCM), a reduction in corneal innervation in patients with DN has been observed.

Copyright of Acta Odontologica Latinoamericana: AOL is the property of Acta Odontologica Latinoamericana and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Arquivos de Neuro-Psiquiatria is the property of Thieme Medical Publishing Inc. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Objetivo. El objetivo de este estudio fue evaluar la adaptación marginal de coronas de disilicato de litio obtenidas mediante técnicas de escaneo (CAD/CAM), antes y después de la cristalización, a través de análisis in vitro con microscopía confocal (MC). Métodos. Fueron confeccionadas 16 réplicas en poliuretano a partir de la pieza 1.4, de modelo typodont, tallada para corona total. Las réplicas fueron divididas en dos grupos, de acuerdo a la técnica de escaneo: Técnica Indirecta (Grupo IND, n=08), donde modelos de yeso fueron escaneados con escáner de laboratorio (inEos X5, Sirona Dental Systems) y Técnica Directa (Grupo DIR, n=08), donde modelos typodont fueron escaneados con escáner intraoral (CEREC BlueCam, Sirona Dental Systems). A seguir, se fresaron (inLab MC XL, Sirona Dental Systems) coronas en disilicato de litio (IPS e.max CAD, Ivoclar Vivadent) y se adaptaron a las réplicas. Se evaluó la adaptación marginal con análisis de MC en dos momentos, antes y después de la cristalización del disilicato de litio. Los datos fueron analizados con la prueba de Mann-Whitney, t de Student y Wilcoxon (α= 0,05). Resultados. Hubo una diferencia estadísticamente significativa en la adaptación marginal horizontal entre los grupos IND y DIR después de la cristalización (p=0,05). En el grupo IND, la comparación de la adaptación marginal vertical antes y después de la cristalización mostró una diferencia estadísticamente significativa (p=0,038). Conclusiones. Las coronas de disilicato de litio obtenidas mediante escaneo directo (CAD/CAM) presentaron menor desajuste marginal vertical. La etapa de cristalización afectó la adaptación marginal de las coronas.

Copyright of Arquivos Brasileiros de Oftalmologia is the property of Arquivos Brasileiros de Oftalmologia and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Cirugía y Cirujanos is the property of Publicidad Permanyer SLU and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Nonalcoholic fatty liver disease (NAFLD) might progress the steatosis to nonalcoholic steatohepatitis (NASH), reaching a cirrhosis state and possibly hepatocellular carcinoma. The liver of three-month-old C57BL/6J mice (wild-type, WT group, n=10) and leptindeficient obese mice (ob/ob group, n=10) were studied, focusing on the mechanisms associated with the activation of the hepatic stellate cells (HSCs) and pro-fibrogenesis. The obese ob/ob animals' liver showed steatosis, increased lipogenesis gene expressions, inflammation, increased pro-inflammatory gene expressions, inflammatory infiltrate, and potential apoptosis linked to a high Caspase 3 expression. In ob/ob mice, liver sections were labeled in the fibrotic zones by anti-alpha-smooth muscle actin (alpha-SMA) and anti-Reelin, but not in the WT mice. Moreover, the alpha-SMA gene expression was higher in the ob/ob group's liver than the WT group. The pro-fibrogenic gene expressions were parallel to anti-alpha-SMA and anti-Reelin immunofluorescence, suggesting HSCs activation. In the ob/ob animals, there were increased gene expressions involved with lipogenesis (Peroxisome proliferator-activated receptor-gamma, Cell death-inducing DFFA-like effector-c, Sterol regulatory element-binding protein-1c, and Fatty acid synthase), pro-fibrogenesis (Transforming growth factor beta1, Smad proteins-3, Yes-associated protein-1, Protein platelet-derived growth factor receptor beta), pro-inflammation (Tumor necrosis factor-alpha, and Interleukin-6), and apoptosis (Caspase 3). In conclusion, the results in obese ob/ob animals provide a clue to the events in humans. In a translational view, controlling these targets can help mitigate the hepatic effects of human obesity and NAFLD progression to NASH. [ABSTRACT FROM AUTHOR]

Introdução: A queratose actínica (QA) é lesão pré-maligna que pode progredir para carcinoma espinocelular. O diagnóstico é clínico, dermatoscópico e por microscopia confocal. Atualmente, aborda-se o tratamento do campo cancerizável, abrangendo QAs clinicamente visíveis e subclínicas, sendo a terapia fotodinâmica (PDT) uma opção terapêutica. Objetivo: Avaliar melhora das QAs e campo cancerizável em pacientes submetidos a PDT com luz do dia, com análise clínica, dermatoscópica e por microscopia confocal. Métodos: Foram selecionados dez pacientes, com múltiplas QAs na face. Realizada a PDT utilizando luz do dia com aminolevulinato de metila e feita documentação fotográfica clínica, dermatoscópica e por microscopia confocal antes do tratamento e 60 dias após seu início. Resultados: Dos nove pacientes que completaram o tratamento, oito (88,8%) apresentaram melhora clínica e regressão no grau da QA com uma sessão. Na dermatoscopia, quatro pacientes (44,4%) apresentaram melhora significativa, três pacientes (33,3%) apresentaram melhora parcial e dois pacientes (22,2%) tiveram suas lesões estáveis. Na microscopia confocal, seis (66,6%) pacientes tiveram regressão no grau da lesão. Conclusões: A PDT com luz do dia se mostrou eficaz para tratamento de QAs, apresentando alto grau de tolerabilidade e eficácia, além de bom perfil de segurança.

Bacterial adhesion and biofilm formation on restoration surfaces could lead to secondary caries and even inflammation of pulpal nerve. Finishing and polishing procedures are crucial to form resistance of materials to bacterial adhesion. The aim of the present study is to compare Streptococcus Mutans (S. mutans) adhesion on restorative materials polished with one- or multi-step systems. 2x5 mm disc-shaped samples were prepared from a resin-modified glass ionomer (RMGI), a compomer, a conventional flowable composite and two flowable bulk-fill composites. Specimens of each group were divided into two groups according to polishing systems (n = 9): One-step (OG) or Multi-step (SL) systems. Surface roughness values were examined by profilometry and one sample of each group were examined for bacterial on confocal laser scanning microscope (CLSM). S. mutans counts were calculated by broth cultivation. Results were analyzed with one-way ANOVA and Bonferroni/Dunn tests. Two flowable bulk-fill composites showed superior roughness values than the conventional flowable and RMGI. Specimens polished with OG system had no significant difference among bacterial counts (p>0.05). After polishing with SL system, Tetric Evo Bulk Flow showed significantly the lowest bacterial adhesion followed by the RMGI and the compomer. CLSM images were in consistent with microbiological culture. All tested materials had lower bacterial adhesion when polished with multi-step system. Multi-step systems should be used with flowable bulk-fill composites to have optimum results in terms of lowering bacterial adhesion and improving surface properties. CLSM images supplies accordance with broth culture of S. mutans thus, this method could be useful on detecting bacterial adhesion. [ABSTRACT FROM AUTHOR]

RESUMEN Objetivo: Identificar las características morfológicas del epitelio, el estroma y el endotelio corneal, así como la densidad celular de este último mediante el empleo de la microscopia confocal de la córnea en pacientes diabéticos. Métodos: Se realizó un estudio descriptivo, comparativo, en 90 ojos; 60 de ellos pertenecientes a pacientes diabéticos (30 tipo 1 y 30 tipo 2) y 30 ojos a pacientes supuestamente sanos. El estudio se realizó en el Instituto Cubano de Oftalmología “Ramón Pando Ferrer” entre enero del año 2012 y enero de 2017. Resultados: Predominó el sexo masculino con 66,7 % en los pacientes con diabetes mellitus tipo 1; el sexo femenino en los pacientes con diabetes mellitus tipo 2 (60 %) y aparentemente sanos (56,7 %). En los pacientes con diabetes mellitus tipo 1 fueron más frecuentes las edades entre 45 y 54 años (33,3 %) y entre 55 y 66 años en los pacientes con diabetes mellitus tipo 2 y aparentemente sanos con 60 y 40 % respectivamente. La morfología del epitelio y el estroma corneal fue normal en el 86,7 y 87,3 % respectivamente. Predominaron las alteraciones de la morfología endotelial en pacientes diabéticos tipo 1 (73,3 %), así como el polimegatismo y el pleomorfismo (73,3 y 56,7 % respectivamente) y la densidad celular más baja (2 222,76 células /mm2). Conclusiones: La ausencia de alteraciones morfológicas del epitelio y el estroma corneal y la presencia de polimegatismo y de pleomorfismo fueron los hallazgos más frecuentes.

RESUMEN Objetivo: Identificar las características morfológicas y morfométricas de las capas de la córnea mediante microscopia confocal en pacientes diabéticos. Métodos: Se realizó un estudio descriptivo, comparativo, en 90 ojos, 60 de ellos pertenecientes a pacientes diabéticos (30 tipo 1 y 30 tipo 2) y 30 a pacientes supuestamente sanos. El estudio se realizó en el Instituto Cubano de Oftalmología “Ramón Pando Ferrer” entre enero del año 2012 y enero de 2017. Resultados: El espesor del epitelio, el estroma y el endotelio corneal fue mayor en los ojos de diabéticos tipo 1 con una media de 54,6; 506,7 y 26,7 micras respectivamente. La membrana basal epitelial se observó en el 20 % de los ojos de pacientes con diabetes mellitus tipo 1, y en el 10 % de los diabéticos tipo 2. El grupo de diabéticos tipo 1 mostró uno y dos plexos nerviosos por campo de microscopia confocal para el 33,3 % cada uno. En los diabéticos tipo 2, predominó un plexo nervioso (40 %) y en pacientes sanos predominaron 4 plexos nerviosos (66,7 %). Ambos grupos de pacientes diabéticos presentaron plexos verticales tortuosos (40 % y 53,3 % respectivamente) y plexos nerviosos oblicuos en el grupo de pacientes supuestamente sanos (80 %). Conclusiones: El estudio de la córnea por microscopia confocal en pacientes diabéticos evidencia mayor espesor corneal total y por capas, membrana basal visible, disminución del plexo nervioso sub-basal con disposición vertical y tortuosidad de las fibras.

Introdução: O fotoenvelhecimento e o melasma são queixas dermatológicas frequentes. A microscopia confocal de reflectância (MCR) é técnica recente que pode ser usada para avaliação diagnóstica dessas dermatoses. Objetivos: Avaliar as características da epiderme e derme nas alterações pigmentares da pele fotodanificada e do melasma pela MCR e comparar os achados dessas alterações com a região perilesional. Métodos: Foi realizado estudo-piloto com oito participantes do sexo feminino, com idades variando de 38 a 50 anos, fototipos de II a IV, com diagnóstico clínico de fotodano (n = 4) e melasma (n = 4) na região malar da face. Foram comparadas a espessura do estrato córneo e da epiderme viável, a profundidade das cristas interpapilares e a presença de estruturas hiper-refrativas na região perilesional e lesional. Resultados e Discussão: As alterações pigmentares da pele fotodanificada revelaram padrão morfológico característico do lentigo solar, como aumento na profundidade das cristas interpapilares na região da lesão. Nas voluntárias com melasma, foi possível observar a presença de células dendríticas na epiderme e melanófagos na derme na região da lesão. Todas as voluntárias apresentaram queratinócitos hiper-refrativos na epiderme da região lesional. Conclusões: Considerando o número de pacientes avaliados, foi possível caracterizar e comparar as alterações pigmentares na pele fotodanificada e no melasma.

O diagnóstico das lesões pigmentadas da face é considerado desafiador uma vez que lesões benignas e malignas podem compartilhar características clínicas e dermatoscópicas semelhantes, principalmente em lesões iniciais, sendo muitas vezes difícil de identificar as lesões de lentigo maligno da face. Assim, a microscopia confocal de reflectância pode se tornar uma ferramenta útil no diagnóstico dos lentigos malignos bem como para o seu planejamento cirúrgico.

The efficient compounds' design taking advantage of the molecule's structural characteristics and efficient search for therapeutic targets has provided effective tools for the research of new treatments when this is focused on disease cellular mechanisms. Phenotypic changes produced by in vitro interaction between molecules and targets can be monitored quantitatively by live cell imaging. To guarantee adequate interaction, it is necessary to consider different crucial elements: 1. Structural characteristics and molecular dynamics of the evaluated compound. 2. Target relevance for the concern physiopathology. However, overview's ignorance of the drug discovery, from structural and cellular problems, has slowed the new treatments research. This literature review presents some important structural aspects for compounds' characterization as therapeutic candidates and experimental approaches for cellular systems development. Subjects discussed are focused on live cell imaging and we also provide relevant examples. Phenotypic monitoring of interactions' produced effects between the chemical candidate and therapeutic target in a cellular system can favor the efficient search of potentially therapeutic molecules. [ABSTRACT FROM AUTHOR]

La meibografía es una tecnología que nos permite evaluar la morfología de las glándulas de Meibomio in vivo de una forma no invasiva y con pocas o nulas molestias. Nuevas tecnologías en el mercado nos permiten realizarla sin necesidad de tocar al paciente y con la ayuda de programas computacionales es posible evaluar de forma objetiva y cuantitativa la pérdida de las mismas. Otras tecnologías de imagenología como la microscopia confocal pueden además analizar el tejido celular adyacente a las glándulas de Meibomio para monitorizar la respuesta al tratamiento instituido. La presente revisión analiza la meibografía desde sus inicios y los avances más recientes en esta área.

The cornea is the tissue of the eye that provides most of the refractive power. Any disease that alters its morphological characteristics affects vision. This is why the early and successful diagnosis of corneal dystrophies guarantees that patients are treated on time to improve their health and quality of life. Confocal microscopy has become the technology per excellence for the detailed study of the cornea in a non-invasive way.

Introduction and Objectives: We aimed to investigate whether Sjögren's syndrome (SS) had distinctive findings in tongue confocal microscopy in a non-invasive manner., Materials and Methods: This retrospective case-control study evaluated corneal and tongue confocal microscopy findings of the right eyes of 25 patients with aqueous deficient dry eye and 12 healthy volunteers without dry eye findings. There were a total of 14 patients diagnosed with SS-associated dry eye (SSDE), while 11 cases were evaluated as non-Sjögren dry eye (NSDE)., Results: A significant difference was observed in the dendritic cell count at the corneal subbasal nerve level between the SSDE and NSDE groups (P=.018). In SSDE group, the confocal microscopy images of dendritiform hyperreflective inflammatory cells in the tongue mucosa were in favor of inflammation. However, these findings were not found in patients with NSDE or in controls., Conclusions: This study showed that confocal microscopy provided a non-invasive evaluation of the inflammatory cells in the tongue of SS patients., (Copyright © 2023 Sociedad Española de Oftalmología. Published by Elsevier España, S.L.U. All rights reserved.)

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El estudio de las interconexiones que establecen las neuronas entre sí o con las células de la glía es uno de los mayores desafíos de la neurociencia. Los anticuerpos fluorescentes permiten la identificación y localización de diferentes estructuras tisulares. Sin embargo, su uso eficiente requiere de la utilización de cortes delgados de muestras, que constituyen una limitación para el estudio de las interconexiones en el sistema nervioso. Asimismo, un mayor espesor de la muestra limita la penetración de la luz emitida por el microscopio, mientras que la opacidad propia del tejido nervioso debida a su alto contenido lipídico dificulta la adquisición de las imágenes al restringir la resolución de los objetos. La técnica CLARITY (del inglés, Clear, Lipidexchanged, Acrylamidehybridized Rigid, Imaging/Immunostaining/In situ hybridizationcompatible, Tissue hYdrogel) permite subsanar estos inconvenientes. Esta técnica fue adaptada en nuestro laboratorio para el estudio de la médula espinal de rata. De acuerdo con el procedimiento realizado, pudimos obtener un órgano completamente translúcido, estructuralmente intacto y que permitió su procesamiento mediante técnicas de inmunofluorescencia y de lectinhistoquímica sin mayores dificultades. A partir de muestras de más de 1 mm de espesor se obtuvieron imágenes confocales de gran resolución y de mayor penetrabilidad que las que se obtienen utilizando las técnicas de procesamiento convencionales. La implementación de esta técnica en nuestro laboratorio permitirá optimizar la información obtenida a partir de muestras de interés.

Cutaneous leishmaniasis may mimic other infections in overlapping endemic areas and timely treatment prevents dissemination of the parasite. The required histopathological and microbiological examinations are not always available, and can only give a deferred confirmation of the diagnosis. In contrast, reflectance confocal microscopy (RCM) allows real-time visualization till the level of papillary dermis. A 59-year-old Brazilian male presented with ulcerated plaques and tumors on the extremities. The clinical differential diagnosis included leishmaniasis and other infections with lymphocutaneous pattern of dissemination. RCM showed the characteristic picture of «eggs in a bird’s nest» which has been described in cutaneous leishmaniasis. The diagnosis of leishmaniasis was later confirmed by skin biopsy, in which Leishmania guyanensis was identified by parasitological examination. After treatment with liposomal amphotericin B, reassessment with RCM corroborated the clinical cure, showing an «empty nest» picture. In conclusion, RCM noninvasively provides useful information for diagnosis and follow-up of cutaneous leishmaniasis. [ABSTRACT FROM AUTHOR]

Copyright of Revista Médica Clínica Las Condes is the property of Editorial Sanchez y Barcelo and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Arquivos Brasileiros de Oftalmologia is the property of Arquivos Brasileiros de Oftalmologia and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Analecta Veterinaria is the property of Universidad Nacional de La Plata and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Implant Dentistry is the property of Lippincott Williams & Wilkins and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Se realizó un estudio descriptivo y transversal de 20 pacientes (20 ojos) en quienes se practicó trasplante perforante de córnea, con evolución favorable, en el Centro Oftalmológico del Hospital General Docente "Dr. Juan Bruno Zayas Alfonso" de Santiago de Cuba, de enero a junio del 2014, y luego se les examinó con microscopia confocal, a fin de describir las características del tejido corneal. Entre las variables seleccionadas figuraron: edad, sexo y causa de indicación de la queratoplastia penetrante; en tanto, los resultados preponderantes fueron: grupo etario de 60-69 años, sexo masculino y queratopatía vesicular como principal causa del trasplante. Asimismo, las alteraciones más frecuentemente halladas en la microscopia confocal resultaron: disminución del número de células endoteliales, aumento en el polimegatismo y alteración en el pleomorfismo endotelial

Copyright of Dental Press Endodontics is the property of Dental Press International and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Introducción: la Cirugía Refractiva es una subespecialidad de la Oftalmología, encargada de los procedimientos quirúrgicos dirigidos al tratamiento de los defectos refractivos. Objetivos: explicar los principios generales de funcionamiento de la microscopia confocal de la córnea, describir las características microscópicas de la córnea normal por microscopia confocal y enunciar los aspectos teóricos generales relacionados con la tecnología láser excimer. Desarrollo: la microscopia confocal es un método no invasivo para el estudio de imágenes microscópicas en tejidos vivos, donde la iluminación y la detección se encuentran en el mismo plano focal; además, la luz se refleja y pasa a través de un segundo lente objetivo. El láser excimer utiliza pulsos controlados de energía de luz ultravioleta de 193 nm de longitud de onda para aplicar ablación sobre el tejido estromal, lo cual produce un patrón de excisión refractivo y genera nuevos radios de curvatura. Asimismo, el término excimer es una forma química diatómica singular que solo existe en estado excitado y durante un tiempo minúsculo. Conclusiones: el estudio de la córnea por microscopia confocal permite la diferenciación de las subcapas del epitelio, el plexo nervioso subbasal, los queratocitos y nervios del estroma, así como las células del endotelio corneal. El láser excimer, posibilita el moldeamiento de la curvatura corneal, con vistas a lograr un resultado refractivo con mínimas alteraciones al tejido circundante; por tanto, a escala internacional, se ha convertido en la técnica quirúrgica prevaleciente para el tratamiento de las ametropías.Introduction: refractive surgery is a subspecialty of the ophthalmology, responsible for the surgical procedures aimed at treating refractive defects. Objectives: to explain the general principles of functioning of the corneal confocal microscopy, to describe the microscopic features of normal cornea by confocal microscopy and to enunciate general theoretical aspects related to the excimer laser technology. General principles: confocal microscopy is a non-invasive method for studying microscopic images in living tissues, where the lighting and detection are in the same focal plane; moreover, the light is reflected and passes through a second objective lens. The excimer laser uses controlled ultraviolet light power pulses of 193 nm wavelength to apply ablation on the stromal tissue, which produces a refractive excision pattern and generates new curvature radii. Also, the term excimer is a single diatomic chemical form that only exists in excited state for a short time. Conclusions: the study of the cornea by confocal microscopy allows the differentiation of the epithelial sublayers, subbasal nervous plexus, keratocytes and stromal nerves, as well as corneal endothelial cells. The excimer laser facilitates the shaping of the corneal curvature to achieve a refractive outcome with minimal disruption to the surrounding tissue, so internationally it has become the prevailing surgical technique for the treatment of ametropies.

Schnyder's corneal dystrophy (SCD) is a rare corneal condition characterized by cholesterol and phospholipids deposition in the stroma and Bowman's layer. We present a case report of a patient who had a progressive corneal stromal haze in both eyes since he was 15 years old. Etiological diagnosis of SCD was well established by In Vivo Confocal Microscopy (IVCM). [ABSTRACT FROM AUTHOR]

Objetivo: describir las modificaciones en la estructura celular corneal después del tratamiento con crosslinking en pacientes con queratocono. Métodos: la muestra estuvo conformada por 30 pacientes (30 ojos) con queratocono progresivo a los que se les realizó crosslinking del colágeno corneal y microscopia confocal. Se analizaron como variables las alteraciones en los plexos nerviosos sub-basal y subepitelial, en los nervios estromales, las modificaciones de los queratocitos en estroma anterior y medio, los cambios de las células endoteliales, la profundidad del tratamiento, así como la presencia y evolución del haze y el edema estromal. Resultados: los plexos nerviosos sub-basal y subepitelial estuvieron ausentes durante el primer mes, hubo una recuperación parcial a los 6 meses (73,3 %) y total al año. Las fibras nerviosas estromales estuvieron ausentes al mes, a los 6 meses se recuperaron en 96,7 % y de manera total al año. Se encontró alteración de la morfología de los queratocitos del estroma anterior y medio en el primer mes (96,7 %), con inicio de la repoblación celular en 83,3 % a los 3 meses y al año se evidenció una regeneración total. El edema estromal estuvo presente en el primer mes en 83,3 % de los casos, solo en 3,3 % a los 6 meses y al año no hubo ningún paciente. El haze se observó al mes y a los tres meses en el 100 % de los pacientes, predominó al mes en el grado II, a los seis meses con grado I y disminuyó a 60 % sin haze al año. No se encontraron alteraciones en la morfología o densidad de las células endoteliales. Conclusiones: los resultados obtenidos reafirman los criterios de que el crosslinking del colágeno corneal es un método de tratamiento seguro.Objective: to describe the changes of the corneal cellular structure of patients with keratoconus after the cross-linking treatment. Methods: the sample was made up of 30 eyes from 30 patients, who had progressive keratoconus and were treated with corneal collagen crosslinking and confocal microscopy. The analyzed variables were the alterations in the sub-basal and sub-epithelial nervous plexuses, in the stromal nerves, the changes in keratocytes in the anterior and medium stroma, the changes of the endothelial cells, the depth of treatment as well as the presence and the progression of haze and the stromal edema. Results: there no sub-basal and sub-epithelial nervous plexuses during the first month, there were re-innervations after 6 months in 73,3 % of eyes and total recovery after 12 months. Likewise, there was no stromal nervous fibers in the first month, but after 6 months they were recovered in 96,7 % of patients and the total recovery was attained after a year. Rarefaction of the keratocytes in the anterior and the medium stroma was found during the first month in 96,7 % of cases, the onset of the cell repopulation was observed after 3 months in 83,3 % of cases and a total regeneration was evident after 1 year. The stromal edema was present in the first month in 83,3 % of cases, 6 months later was seen in 3.3 % and after 1 year, no patient was affected. Haze was identified on the first month and after 3 months in 100 % of patients. The haze grade II predominated in the first month, the grade I after 6 months and 60 % of patients presented with no haze after a year. Conclusions: the obtained results confirmed the criteria which establish that the corneal collagen crosslinking is a safe treatment method.

OBJETIVOS: Avaliar a concordância entre as técnicas de microscopia de polarização e microscopia confocal na avaliação do fuso meiótico de oócitos humanos maturados in vivo. MÉTODOS: Estudo prospectivo que avaliou oócitos com o primeiro corpúsculo polar extruído obtidos de mulheres inférteis submetidas à estimulação ovariana para realização de injeção intracitoplasmática de espermatozoide. Os oócitos com o primeiro corpúsculo polar extruído foram avaliados por meio da microscopia de polarização e, imediatamente após, foram fixados e corados para avaliação dos microtúbulos e cromatina pela microscopia confocal de alto desempenho. Foram comparadas as técnicas de microscopia de polarização e confocal, de acordo com a visualização ou não do fuso meiótico pela microscopia de polarização e a presença ou não de anomalias meióticas à análise pela microscopia confocal. Foram calculados os intervalos de confiança, o índice de Kappa e a concordância entre as metodologias, considerando a análise da microscopia de imunofluorescência como padrão-ouro para avaliação de normalidade do fuso e distribuição cromossômica oocitária. RESULTADOS: Observou-se que 72,7% dos oócitos em metáfase II com fuso celular não visível à polarização apresentaram anormalidades meióticas à análise confocal e que 55,6% dos oócitos em metáfase II com fuso celular visível à polarização apresentaram-se como oócitos anormais à análise confocal. Somente 44,4% dos oócitos com fuso celular visível à polarização apresentaram-se como normais à análise confocal. A concordância entre os métodos foi de 51,1% (Kappa: 0,11; IC95% -0,0958 - 0,319). CONCLUSÕES: A baixa concordância entre a microscopia de polarização e a confocal na avaliação do fuso meiótico oocitário sugere que a visualização do fuso meiótico de oócitos humanos em metáfase II pela microscopia de polarização tem limitado o valor preditivo de normalidade meiótica oocitária.PURPOSE: To evaluate the concordance between polarization microscopy and confocal microscopy techniques in the evaluation of the meiotic spindle of human oocytes matured in vivo. METHODS: Prospective study that evaluated oocytes with the first polar extruded body obtained from infertile women who had undergone ovarian stimulation for intracytoplasmic sperm injection. The oocytes with the first polar extruded body were evaluated by polarization microscopy and were then immediately fixed and stained for microtubule and chromatin evaluation by high-performance confocal microscopy. We determined the correlation of polarization microscopy with confocal microscopy in the detection of meiotic oocyte anomalies, and we also evaluated the percentage of oocytes with a visible and non-visible cell spindle by polarization microscopy and with meiotic normality and abnormalities by confocal microscopy. Confidence intervals, Kappa's index and concordance between the methodologies were calculated, considering immunofluorescence microscopy analysis as the golden-standard for evaluating normal spindle and oocyte chromosome distribution. RESULTS: We observed that 72.7% of metaphase II oocytes with a nonvisible meiotic spindle by polarization microscopy showed no meiotic abnormalities by confocal analysis and 55.6% of metaphase II oocytes with a visible meiotic spindle by polarization microscopy were found to be abnormal oocytes by the confocal analysis. Only 44.4% of oocytes with a visible meiotic spindle by polarization microscopy were found to be normal by confocal analysis. Concordance between the methods was 51.1% (Kappa: 0.11; 95%CI -0.0958 - 0.319). CONCLUSIONS: The low correlation between polarization microscopy and confocal microscopy in the assessment of oocyte meiotic spindle suggests that visualization of the meiotic spindle of human oocytes at metaphase II by polarization microscopy is not a good indicator of oocyte meiotic normality.

Introducción: el análisis de las variables histológicas en pacientes miopes se ha convertido en los últimos años en elemento primordial para asegurar la óptima calidad de indicación y seguimiento en pacientes con defectos refractivos sometidos a tratamiento quirúrgico con láser de excímeros. Objetivo: determinar la densidad de queratocitos en pacientes miopes. Método: se realizó un estudio observacional, descriptivo, transversal con una muestra de 30 ojos (15 pacientes) miopes que cumplieron con los criterios establecidos en las Normas y Procedimientos de Oftalmología para el tratamiento con láser de excímeros, de enero a marzo de 2011 en el Centro Oftalmológico de Pinar del Río. Se utilizó el microscopio confocal ConfoScan 4 de NIDEK para la obtención y estudio de las imágenes in vivo de tejido corneal. Los pacientes otorgaron su consentimiento para la participación en el estudio. Resultados: se obtuvo densidad de queratocitos de 21,33 cel/mm3 en estroma anterior; 20,12 cel/mm3 en estroma medio y 18,46 cel/mm3 en estroma posterior. No se obtuvieron diferencias significativas según la magnitud de la ametropía. Conclusiones: la densidad de queratocitos fue mayor en estroma anterior y menor en estroma posterior. La densidad de queratocitos se mantuvo en valores muy similares independientemente de la magnitud de la ametropía.

Objetivo: Describir las alteraciones morfológicas de la cicatrización estromal y la reinervación por microscopia confocal en pacientes operados de LASIK posterior a queratotomía radial de 15 años de evolución. Métodos: Se realizó un estudio observacional, descriptivo y longitudinal prospectivo, en 25 pacientes (50 córneas) operados de queratotomía radial no complicados y reintervenidos con LASIK. Se estudiaron las características de la cicatrización estromal y la regeneración de fibras nerviosas corneales mediante microscopia confocal de la córnea con Confoscan S4. Se emplearon medidas de resumen de estadística descriptiva. Resultados: La cicatrización estromal se limitó al borde del colgajo. Hubo un aumento de la actividad queratocitaria en el lecho estromal en las primeras semanas del postoperatorio y disminución de la densidad queratocitaria por apoptosis observada hasta el año de evolución. En el preoperatorio de LASIK el número de plexos nerviosos se encontraban normales en más del 80 % de las córneas. En los primeros tres meses posoperatorios existió ausencia de reinervacion, su recuperación comenzó entre el sexto y octavo mes, con evidente alteración de la morfología y cantidad de plexos nerviosos por campo de microscopia confocal al año. Conclusiones: Las características de la cicatrización estromal estudiadas evolucionaron de manera similar a lo descrito en pacientes que no presentaban dicha queratotomía radial como antecedente. La regeneración nerviosa comenzó a partir de los seis meses con la aparición de los nodos nerviosos y un aumento progresivo de la sensibilidad corneal.Objective: To describe the morphological alterations in the stromal scaring and re-inervation by using confocal microscopy in patients operated on with LASIK after radial postkeratotomy of 15 years old evolution. Methods: A prospective, longitudinal, observational and descriptive study was conducted in 25 patients (50 corneas) operated on by radial keratotomy, with no further complications and re-operated on with Laser in Situ keratomileusis (SHWIND Esiris). The characteristics of the corneal structures and the regeneration of corneal nervous fibers were studied by means of confocal microscopy with Confoscan S4. Summary measures were used. Results: The stromal scaring was limited on the edge of the flap; there was an increase of the keratocyte activity in the stromal channel during the first weeks of the postoperative period and a decrease of the keratocyte density caused by apoptosis observed up to one year of evolution. Before Lasik, the number of nervous plexuses was normal in more than 80% of the corneas with incisional corneal surgery. In the first 3 postoperative months there was no nervous regeneration; but this process began between 6th and 8th months, with evident alteration of the morphology and quantity of nervous plexuses per field of confocal microscopy a year after surgery. Conclusion: The evolution of the microscopic characteristics of the cornea, which were studied with microscopy confocal in patients operated on by Lasik after radial keratotomy, was similar to that described in patients who had not undergone this type of surgery. The nervous regeneration began after 6 months when nervous nodes and progressive increase of corneal sensitivity occurred.

Objetivo: Analizar las estructuras celulares por microscopia confocal, Confoscan 4, en córneas sanas en nuestro medio. Métodos: Se realizó un estudio prospectivo longitudinal a 100 ojos sanos de médicos que trabajan en nuestra institución, y pacientes que asistieron al servicio de córnea. Esta investigación fue desde mayo de 2007 a mayo 2008, en el Instituto Cubano de Oftalmología "Ramón Pando Ferrer", La Habana. En los médicos se examinaron ambos ojos y en los pacientes el ojo no afectado. Se recopilaron un total de 50 casos sin afección corneal. Resultados: De los 100 ojos estudiados, 64 tenían paquimetrías por encima del valor medio. Estuvieron presentes los tres tipos de células epiteliales en casi la totalidad de los pacientes; así como los queratocitos en las diferentes profundidades del estroma corneal. La mayoría de los ojos tenían un conteo celular endotelial por encima de 2 500, cifra comprendida dentro de los valores normales. Se encontraron fibras nerviosas en cada una de sus capas. Conclusiones: La microscopia confocal se presenta como una nueva herramienta que permite observar en vivo la histología corneal y complementar las observaciones de la biomicroscopia convencional. Esto constituye un reto para el mejor entendimiento de la histopatología corneal. De esta manera podemos actuar de forma profiláctica y terapéutica, en el seguimiento y evolución de patologías corneales.Objective: This paper is aimed at analyzing the corneal cellular structures through Confoscan S4-aided confocal microscopy in apparently healthy corneas. Methods: A prospective longitudinal study of 100 healthy eyes from practicing doctors, and from patients who had attended the corneal service at “Ramón Pando Ferrer” Cuban Institute of Ophthalmology in Havana since May 2007 was conducted. Both eyes of participating doctors were examined whereas the non-affected eye was examined in the patients. A total of 50 cases with no corneal disease notified from May 2007 to May 2008 were considered. Results: The 100 studied eyes came from apparently healthy patients, and the pachymetry values of 64 of them were above the average. Since they were healthy corneas, the 3 types of epithelial cells as well as the keratocytes were present in almost all the patients and at different depths of the corneal stroma, respectively. Most of the studied eyes had endothelial cell counts above 2 500, a figure within the normal range, and there were nerve fibers in each of their layers. Conclusion: Confocal microscope is a new tool that allows in vivo observation of corneal histology and thus supplements conventional biomicroscopy observations. It contributes to better understanding of corneal pathology, with a view to prophylactically and therapeutically acting upon corneal pathologies during their follow-up and evolution.

Objetivo: Describir los hallazgos morfométricos del endotelio corneal por microscopia confocal con CONFOSCAN S-4. Métodos: Estudio descriptivo transversal de 102 ojos con queratocono en el período de septiembre de 2008 a septiembre 2009. A estos pacientes se les realizó microscopia confocal con CosfoscanS-4 para el estudio del endotelio corneal atendiendo el grado de queratocono. Se analizó el comportamiento de la evolución del queratocono según edad y sexo. Las imágenes fueron analizadas y procesadas mediante un programa informático diseñado específicamente para esto. Resultados: Fueron semejantes las edades de los pacientes con queratocono grado I y II, (35,2 y 34,7 años), los grado III presentaron una edad promedio mayor (38,4 años), sin diferencias significativas (p= 0,279). El sexo femenino predominó en 80,4 % de los pacientes. El 100 % de los queratoconos grado III tuvieron endotelios patológicos. Los valores promedios de la densidad celular en los queratoconos grado III (2585,9 células/mm²) resultó no significativo (p= 0,339). El polimegatismo en los queratoconos grado III para un 48,69 % fue significativo (p= 0,002). En el pleomorfismo resultó significativo las diferencias observadas entre los tres grados (p= 0,002). Conclusión: Predominó el queratocono grado II para las mujeres y el grado I para los hombres. Los hallazgos morfológicos se manifestaron en la forma y tamaño de las células endoteliales. En córneas con queratocono grado II y III confluyeron células de mediano y gran tamaño con pérdida de su hexagonalidad. La densidad celular se mantuvo dentro del rango de valores normales para cualquier grado de queratocono.Objective: To describe the morphometric findings of the corneal endothelium confocal microscopy with CONFOSCAN S-4 Methods: Descriptive cross-sectional study of 102 eyes with keratoconus performed from September 2008 to September 2009. The study patients had undergone confocal microscopy with CONFOSCAN S-4 in order to study their corneal endothelium according to the keratoconus condition. The evolution of keratoconus was analyzed in terms of age and sex. Tailor-made software made the analysis and processing of images possible. Results: The ages of patients with keratoconus grades I and II, (35,2 and 34,7 years) were pretty similar whereas the average age of keratoconus grade III patients (38,4 years) was higher, but no significant differences were found (p=0,279). Females prevailed (in our study accounting for 80,4%. One hundred percent of keratoconus grade III presented with pathological endothelium. The values averages of the cell density in the keratoconus grade III (2585,9 cell/mm2) it was not significant (p= 0,339). The polymegathism of 48, 69% in the keratoconus grade III were significant (p= 0,002). As for the pleomorphism, the differences were statistically significant among the three grades (p= 0,002) Conclusions: Keratoconus grade II prevailed in women and the grade I in men. From the morphological point of view, the morphological findings lied in the form and size of the endothelial cells. Keratoconus grade II and II showed a combination of medium and large cells and loss of hexagonal shape. The cell density was within the normal range of values for any keratoconus grade.

Objetivo: Determinar los cambios estructurales de la córnea en la cirugía de catarata por facoemulsificación sin complicaciones. Métodos: Se realizó un estudio prospectivo de pacientes operados de catarata por facoemulsificación coaxial por la técnica de pre chop sin complicaciones. A estos se les realizó microscopia confocal de la córnea con el CONFOSCAN 4 (Nidek Technologies) con el objetivo de 40x y adaptador Z-Ring. Se realizó el estudio en el preoperatorio y en el posoperatorio (a las 24 horas, después de una semana, de un mes y a los tres meses). Resultados: Se demostraron cambios estructurales en la córnea como células epiteliales con núcleos hiperreflectivos alargadas en ocasiones y áreas de hiperreflectividad anómala a las 24 horas del posoperatorio. Persistieron queratocitos activados y la disminución de la hiperreflectividad de la matriz extracelular que desapareció al mes. Conclusiones: Aunque por biomicroscopia no se observen alteraciones corneales en el posoperatorio de la cirugía de catarata por facoemulsificación, sí se pueden demostrar por microscopia confocal de la córnea. Estas variaciones no influyen en la recuperación visual óptima de los pacientes.Objective: To determine the structural changes in the cornea in the cataract surgery using phacoemulsification without complications. Methods: A prospective study of patients operated on from cataract using the coaxial phacoemulsification (Pre Chop) technique without complications was carried out. These patients also underwent confocal microscopy of the cornea with Confoscan4 (Nidek Technologies) with 40x target and Z - Ring adapter. The study was performed in the preoperative period and postoperative period for 24 hours, one week, one month and three months after surgery. Results: Structural changes were observed in the cornea such as epithelial cells with hypereflectivity nucleus, occasionally elongated, , areas of anomalous hypereflectivity 24 hours after surgery. Activated Keratocytes persisted as well as decreased hypereflectivity of the extracellular matrix that disappeared after a month. Conclusions: Although biomicroscopy did not show corneal alterations in the postoperative period of the cataract surgery using phacoemulsification, these can be observed in confocal microscopy of the cornea. These variations do not have an impact on optimal visual recovery of the patients.

OBJETIVO: A granulomatose de Wegener (GW) pode causar dano nas células endoteliais e fenômenos tromboembólicos. Entretanto, poucos estudos analisaram a microcirculação pulmonar - artérias pulmonares de pequeno/médio calibre (APPMC) - em pacientes com GW. O objetivo deste estudo foi quantificar trombos de fibrina em amostras de APPMC de pacientes com GW. MÉTODOS: Analisamos 24 APPMC de seis pacientes com GW e 16 APPMC de quatro pacientes controles sem WG. Utilizamos CD34 para a marcação do endotélio em todas as amostras e microscopia confocal a laser para detectar trombos de fibrina intravasculares. Calculamos a área total do vaso, a área livre do lúmen e a área trombótica. RESULTADOS: A média da área total do vaso foi similar no grupo GW e no grupo controle (32.604 µm² vs. 32.970 µm², p = 0,8793). Trombos foram identificados em 22 das 24 APPMC (91,67%) no grupo GW, e em nenhuma do grupo controle (p < 0,0001; OR = 297 (IC95%: 13,34-6.612). A média da área trombótica foi maior no grupo GW do que no grupo controle (10.068 µm² vs. 0.000 µm², p < 0,0001). Em contraste, a média da área livre do lúmen foi menor no grupo GW que no grupo controle (6.116 µm² vs. 24.707 µm², p < 0,0001). CONCLUSÕES: A microscopia confocal a laser mostrou uma associação significante entre trombose microvascular pulmonar e GW. Isso sugere um possível papel da trombose microvascular na fisiopatologia da GW pulmonar, evocando o potencial benefício da anticoagulação na GW pulmonar. Entretanto, novos estudos são necessários para confirmar nossos achados, assim como um ensaio clínico randomizado a fim de testar o papel da anticoagulação no tratamento de pacientes com GW pulmonar.OBJECTIVE: Wegener's granulomatosis (WG) can cause endothelial cell damage and thromboembolic events. Nevertheless, there have been few studies on the pulmonary microcirculation-small and medium-sized pulmonary arteries (SMSPA)-in patients with WG. The objective of this study was to quantify fibrin thrombi in the SMSPA of patients with WG. METHODS: We analyzed 24 SMSPA samples collected from six patients with WG and 16 SMSPA samples collected from four patients without WG. In all samples, we used the endothelial cell marker CD34 and confocal laser scanning microscopy in order to detect intravascular fibrin thrombi. We calculated the total vessel area, the free lumen area, and the thrombotic area. RESULTS: The mean total vessel area was similar in the WG and control groups (32,604 µm² vs. 32,970 µm², p = 0.8793). Thrombi were present in 22 (91.67%) of the 24 WG group samples and in none of the control group samples (p < 0.0001; OR = 297; 95% CI: 13.34-6,612). The mean thrombotic area was greater in the WG group samples than in the control group samples (10,068 µm² vs. 0.000 µm²; p < 0.0001). In contrast, the mean free lumen area was smaller in the WG group samples than in the control group samples (6,116 µm² vs. 24,707 µm²; p < 0.0001). CONCLUSIONS: Confocal laser scanning microscopy revealed a significant association between pulmonary microvascular thrombosis and WG. This suggests a possible role of microvascular thrombosis in the pathophysiology of pulmonary WG, evoking the potential benefits of anticoagulation therapy in pulmonary WG. However, further studies are needed in order to confirm our findings, and randomized clinical trials should be conducted in order to test the role of anticoagulation therapy in the treatment of patients with pulmonary WG.