Value of multi-modality small fiber assessments in a genotypically diverse cohort of transthyretin-related amyloidosis in the early stages of disease.

INTRODUCTION: Transthyretin-related amyloidosis (ATTRv) is a progressive multisystem disorder, predominantly involving the peripheral nerve system (PNS) and heart. Quantification of small fiber damage may help guide treatment decisions, as amyloid deposits frequently affect those fibers early in disease course. Corneal confocal microscopy (CCM) is a promising method to monitor patients with ATTRv, due to similarities between corneal nerves and PNS, as the cornea is innervated by Aδ and C fibers. METHODS: We compared CCM measures from ATTRv patients to a group of healthy individuals, matched by age and gender. We then investigated the correlations between small fiber tests (SFT): CCM, LDI-Flare and CDT, COMPASS-31 and disability scales (RODS and ONLS) in patients. RESULTS: Of 20 patients (6 with V30M), mean age 50.3±15.3Y, 7 female (35%), six (30%) had polyneuropathy and 10 (50%) carpal tunnel syndrome. CDT was abnormal in 9 and LDI-flare in 6 patients. CCM was abnormal in 19 tested patients and significantly reduced when compared to controls (CNFL: 6.31±0.31 vs. 15.21±1.02mm/mm2, p<0.001). Mean COMPASS-31-scores were 22.27±22.84; RODS and ONLS were 38.15±12.33 and 2.05±2.3, with no significant differences between sub-group scores. Disease duration was significantly correlated with ONLS (0.43, p=0.05) and RODS (0.46, p=0.03). There were no significant correlations between measures of disability and SFT. CONCLUSIONS: In a diverse cohort of ATTRv patients, CCM was the most frequent abnormal measurement. CCM can be a useful test to triage patients in the early disease stages and with few or equivocal symptoms.

Reflectance Confocal Microscopy Terminology in Spanish: A Delphi Consensus Study. / Consenso de terminología en microscopia confocal de reflectancia en español mediante método Delphi.

The terminology used to describe reflectance confocal microscopy (RCM) findings in both melanocytic and nonmelanocytic lesions has been standardized in English. We convened a panel of Spanish-speaking RCM experts and used the Delphi method to seek consensus on which Spanish terms best describe RCM findings in this setting. The experts agreed on 52 terms: 28 for melanocytic lesions and 24 for nonmelanocytic lesions. The resulting terminology will facilitate homogenization, leading to a better understanding of structures, more standardized descriptions in clinical registries, and easier interpretation of clinical reports exchanged between dermatologists.

Corneal ectasia following photorefractive keratectomy: a confocal microscopic case report and literature review / Ectasia corneana após ceratectomia fotorrefrativa: relato de caso de microscopia confocal e revisão da literatura

ABSTRACT The occurrence of corneal ectasia after photorefractive keratectomy is a rare but serious complication of refractive surgery. Possible risk factors are not well assessed, but a probable reason is the failure to detect keratoconus preoperatively. In this report, we describe a case of corneal ectasia after photorefractive keratectomy in a patient who presented a suspicious tomography pattern preoperatively but had no degenerative alterations associated with pathologic keratoconus, as revealed by in vivo corneal confocal microscopy. We also review eligible case reports of post-photorefractive keratectomy ectasia to find similar characteristics.

RESUMO 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.

A novel agent for myeloma causing toxic keratopathy, belantamab mafodotin: a case report and literature review / Belantamabe-mafodotina, um novo agente para mieloma, causando ceratopatia tóxica: relato de caso e revisão da literatura

ABSTRACT A 60-year-old-male with refractory relapsed multiple myeloma presented with redness, pain, foreign body sensation, and blurred vision in both eyes that gradually increased after his third belantamab mafotodin infusion. Biomicroscopy revealed bilateral microcyst-like epithelial changes and epithelial crystal-like deposits, whereas in vivo confocal microscopy revealed intraepithelial and subepithelial hyperreflective deposits in corneal epithelium. Belantamab mafodotin therapy was discontinued for seven weeks due to corneal toxicity, which cleared progressively. We aim to demonstrate belantamab mafodotin-related corneal toxicity that may be detected using slit lamp and in vivo confocal biomicroscopy.

RESUMO Um homem de 60 anos, diagnosticado com mieloma múltiplo recidivante refratário, apresentou vermelhidão, dor, sensação de corpo estranho e visão turva em ambos os olhos, aumentando gradualmente após sua terceira infusão de belantamabe mafodotina. À biomicroscopia, foram observadas alterações epiteliais bilaterais semelhantes a microcistos e depósitos epiteliais semelhantes a cristais. A microscopia confocal in vivo revelou depósitos hiper-refletivos intraepiteliais e subepiteliais na córnea. Devido à toxicidade corneana, a terapia com belantamabe mafodotina foi interrompida por sete semanas e a toxicidade foi gradualmente resolvida. Nosso objetivo é demonstrar os achados à biomicroscopia confocal in vivo e à lâmpada de fenda da toxicidade corneana relacionada ao belantamabe mafodotina.

Universal adhesives applied to deep dentin with different bonding treatments / Adhesivos universales aplicados a dentina profunda con diferentes tratamientos de unión

ABSTRACT Eighth-generation adhesives may be applied with total etch, selective-etch or self-conditioning, and serve as primers for non-dental substrates. Aim: To determine the bonding characteristics of universal adhesives applied to the deep pulp wall with different strategies, by means of shear bond strength and laser microscopy. Materials and Method: Cavities 4 mm deep and maximum width were carved in 36 extracted molars. Nine groups were formed according to dental substrate treatment and adhesives, as follows: Total-etch: group 1-Monobond 7 self-etch, group 2-One coat 7 universal, and group 3-Single bond universal; Adamantine etch: group 4-Monobond 7 self-etch, group 5-One coat 7 universal, and group 6-Single bond universal; Self-conditioning: group 7-Monobond 7 self-etch, group 8-One coat 7 universal, and group 9-Single bond universal. Molars were filled following the manufacturer's instructions. Three specimens per group (27 altogether) were used to determine shear bond strength using a universal testing machine, while layer thicknesses were measured on the remaining specimens using microscope images and Olympus LEXT 3D Software. Analysis of variance was used to compare data. Results: Mean (standard deviation) bond strength in megapascals (MPa) was: group 1: 7.06±3.01; group 2: 10.74±4.36; group 3: 8.20±3.92; group 4: 7.41±2.23; group 5: 6.84±1.50; group 6: 5.86±2.10; group 7: 5.83±1.94; group 8: 7.14±2.37; group 9: 8.06±3.51. Bond strength was higher (p=0.049) for total-etch (8.61±3.96) than for selective etch (6.71±1.98) and self-conditioning (6.91±2.68). No significant difference was found among the three adhesives (p=0.205). Adhesive layer in micrometers (μm) was total-etch 8.71±4.93, selective etch 5.49±1.70 and self-conditioning 6.27±3.01, with no significant difference. Conclusions: There were significant differences among bonding strategies, with the highest values for total-etch. No significant difference was observed between self-conditioning and selective etch. No significant difference was found among the adhesives, which all behaved similarly. The greatest adhesive layer thicknesses were recorded in the total-etch group, with no significant difference among the various adhesive approaches.

RESUMEN Los adhesivos universales de octava generación pueden ser aplicados con diferentes estrategias de unión: grabado total, grabado selectivo o autoacondicionamiento. Además, imprimen sustratos no dentales. Objetivo: Determinar las caracteristicas de unión de adhesivos universales con diferentes estrategias en pared pulpar profunda mediante resistencia adhesiva al corte y microscopía laser. Materiales y Método: En 36 molares se tallaron cavidades de 4 mm de profundidad y ancho máximo. Se dividieron en 9 grupos según tratamientos y adhesivos. Grabado total: grupo 1-Monobond 7 self-etching, grupo 2-One coat 7 universal y grupo 3-Single bond universal; Grabado selectivo: grupo 4-Monobond 7 self-etching; grupo 5-One coat 7 universal y grupo 6-Single bond universal y Autoacondicionamiento: grupo 7-Monobond 7 self-etching; grupo 8-One coat 7 universal y grupo 9-Single bond universal. Las obturaciones se realizaron siguiendo las instrucciones del fabricante. La resistencia adhesiva al corte se determinó utilizando una máquina de ensayo universal sobre 27 especímenes mientras que los restantes fueron empleados para evaluar los espesores de la capa generado sobre imágenes obtenidas con microscopía y con el software Olympus LEXT 3D. Se ultilizó análisis de varianza. Resultados: Resistencia adhesiva en megapascal (MPa) media (desviación estándar): grupo 1: 7,06±3,01; grupo 2: 10,74±4,36; grupo 3: 8,20±3,92; grupo 4: 7,41±2,23; grupo 5: 6,84±1,50; grupo 6: 5,86±2,10; grupo 7: 5,83±1,94; grupo 8: 7,14±2,37; grupo 9: 8,06±3,51. Grabado total (8,61±3,96) registró los valores mayores (p=0,049) en comparación a grabado selectivo (6,71±1,98) y autoacondicionamiento (6,91±2,68). Los adhesivos no tuvieron diferencias significativas (p=0,205). Capa adhesiva en μm: Grabado total (8,71±4,93); grabado selectivo (5,49±1,70) y autoacondicionamiento (6,27±3,01) sin diferencias significativas (p=0,073). Conclusiones: Las estrategias de unión mostraron diferencias significativas; los valores más altos se obtuvieron con grabado total y entre autoacondicionamiento y grabado selectivo no hubo significancia. Los adhesivos evidenciaron comportamientos similares sin registrar diferencias significativas. Los mayores espesores de capa fueron con grabado total sin diferencias significativas entre las técnicas.

Corneal confocal microscopy in patients with distal symmetric polyneuropathy compared to controls / Microscopia confocal de córnea em pacientes com polineuropatia simétrica distal comparados a controles

Abstract 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. Objective To compare, through CCM, diabetic patients with symptomatic distal symmetric polyneuropathy (DSP) and controls. Methods In the present study, through CCM, we compared the morphological changes in the sub-basal epithelial corneal plexus of 35 diabetic patients with symptomatic DSP with 55 controls. Moreover, we sought to determine a pattern of change regarding the severity stages of DSP, comparing the clinical, laboratory, and nerve-conduction (NC) variables. Results Differences between the control and diabetic groups were observed for the following variables, respectively: age (44.9 ± 13.24 years versus 57.02 ± 10.4 years; p< 0.001); fiber density (29.7 ± 10.2 versus 16.6 ± 10.2; p< 0.001); number of fibers (4.76 ± 1.30 versus 3.14 ± 1.63; p< 0.001); number of Langerhans cells (4.64 ± 8.05 versus 7.49 ± 10.3; p= 0.035); tortuosity (p< 0.05); and thickness (p< 0.05). Furthermore, inverse relationships were found regarding fiber density and age (p< 0.01) and fiber density and the severity of the disease (p< 0.05). A positive relationship between the conduction velocity of the fibular nerve and fiber density (p< 0.05) was also observed. Conclusion Corneal confocal microscopy proved to be a fast, noninvasive and reproducible method for the diagnosis, staging, and monitoring of diabetic DSP.

Resumo Antecedentes A neuropatia diabética (ND) é condição clínica muito frequente no mundo inteiro. Os testes diagnósticos atualmente preconizados são pouco sensíveis, como a eletroneuromiografia, ou invasivos, como a biópsia de pele. Novas técnicas de investigação complementares têm sido desenvolvidas a fim de identificar o acometimento precoce do nervo periférico. Com o advento da microscopia confocal de córnea (MCC), observou-se redução da inervação da córnea em pacientes com ND. Objetivo Comparar, por meio da MCC, pacientes diabéticos com polineuropatia simétrica distal (PSD) sintomática e controles. Métodos Neste estudo, por meio da MCC, comparamos as alterações morfológicas do plexo sub-basal epitelial da córnea de 35 pacientes diabéticos com PSD sintomática com 55 indivíduos controles. Além disso, buscamos determinar um padrão de alteração entre os estágios de gravidade da PSD, comparando variáveis clínicas, laboratoriais e de neurocondução. Resultados Diferenças entre os grupos controle e diabéticos foram verificadas com relação às seguintes variáveis, respectivamente: idade (44,9 ± 13,24 anos versus 57,02 ± 10,4 anos; p< 0,001); densidade das fibras (29,7 ± 10,2 versus 16,6 ± 10,2; p< 0,001); número de fibras (4,76 ± 1,30 versus 3,14 ± 1,63; p< 0,001); número de células de Langerhans (4,64 ± 8,05 versus 7,49 ± 10,3; p= 0,035); tortuosidade (p< 0,05), e espessura (p < 0,05). Além disso, relações inversamente proporcionais foram verificadas entre a densidade das fibras e a idade (p< 0,01), e entre a densidade das fibras e a gravidade da doença (p< 0,05). Observou-se ainda uma relação positiva entre a velocidade de condução do nervo fibular e a densidade das fibras (p< 0,05). Conclusão A MCC constitui um método rápido, não invasivo e reprodutível para o diagnóstico, o estadiamento, e o acompanhamento da PSD diabética.

Subbasal corneal nerve damage in patients with bacterial keratitis: in vivo confocal microscopy study / Danos no plexo nervoso subbasal corneano em pacientes com ceratite bacteriana utilizando a microscopia confocal in vivo

ABSTRACT Purpose: To examine subbasal corneal nerve changes in patients with bacterial infectious keratitis using in vivo confocal microscopy. Methods: Thirteen patients (13 eyes) with unilateral bacterial keratitis and 12 healthy controls were prospectively enrolled in the study. In vivo confocal microscopy was performed in all the patients at 2 time points, in the acute phase of infectious keratitis and at 28 ± 0.6 months after resolution of the infection. Results: The subbasal nerve length was 5.15 ± 1.03 mm/mm2 during the acute phase of bacterial keratitis (compared with that of the controls: 19.02 ± 1.78 mm/mm2, p<0.05). Despite the significant corneal nerve regeneration over the interval of 28 months after the resolution of the infection, the nerve density was still significantly reduced as compared with that of the controls (9.73 ± 0.93 mm/mm2, p<0.05). Moreover, in vivo confocal microscopy images showed diffuse high-reflecting areas referring to the scar tissue areas with thin and tortuous nerve branches regenerating toward these areas. Conclusions: A partial corneal nerve regeneration of subbasal nerve plexus during the first 28 months after the acute phase of infectious keratitis was observed. Moreover, the regenerated nerves of the patients remained morphologically altered as compared with those of the healthy controls. These results may be relevant to the clinical follow-up and surgical planning for these patients.

RESUMO Objetivo: Relatar as alterações no plexo nervoso corneano subbasal em pacientes com ceratite infecciosa de origem bacteriana utilizando a microscopia confocal in vivo. Métodos: Treze olhos de 13 pacientes com ceratite bacteriana unilateral e 12 indivíduos saudáveis como grupo controle foram incluídos prospectivamente no estudo. A microscopia confocal in vivo foi realizada em todos os pacientes em 2 momentos: na fase aguda da ceratite infecciosa e após 28 ± 0,6 meses da resolução da infecção. Resultados: A densidade dos nervos no plexo subbasal foi de 5,15 ± 1,03 mm/mm2 na fase aguda da ceratite infecciosa (comparada com o grupo controle: 19,02 ± 1,78 mm/mm2, p<0,05). Apesar de significativa regeneração dos nervos corneanos ao longo de um intervalo de 28 meses após a resolução da infecção, a densidade dos nervos se manteve significativamente reduzida (9,73 ± 0,93 mm/mm2) quando comparada com o grupo controle (19,02 ± 1,78 mm/mm2, p<0,05). Além disso, as imagens obtidas com a microscopia confocal mostraram áreas de hiperreflectividade referente ao tecido corneano cicatricial com ramos de nervos, afinados e tortuosos, se regenerando nessas áreas. Conclusões: Foi observado regeneração parcial dos nervos do plexo corneano subbasal durante os primeiro 28 meses após a resolução da fase aguda da ceratite infecciosa. Além disso, os nervos corneanos regenerados se mantiveram morfologicamente alterados quando comparados ao grupo controle. Esses resultados podem ser relevantes para o acompanhamento clínico e planejamento cirúrgico desses pacientes.

Endotelitis por citomegalovirus y su diagnóstico mediante la microscopia confocal / Cytomegalovirus endotheliitis and its diagnosis by confocal microscopy

Objetivo: Describir las características del endotelio corneal por microscopia confocal en pacientes con lesiones basofílicas de inclusión como patognomónicas de endotelitis por citomegalovirus. Métodos: Se realizó un estudio observacional de casos clínicos en pacientes con diagnóstico de endotelitis por citomegalovirus, atendidos en la Consulta de Córnea del Instituto Cubano de Oftalmología "Ramón Pando Ferrer", entre febrero del año 2010 y junio del 2018. La muestra incluyó 39 pacientes con diagnóstico clínico de endotelitis, a quienes se les realizó microscopia confocal con el equipo Confoscan 4 (Nidek Technologies). Resultados: De los 39 pacientes, 29 fueron del sexo masculino y 10 del femenino. Todos tenían entre 41 y 60 años de edad. En el 97,4 por ciento de los casos existió el antecedente de una conjuntivitis viral y un solo paciente fue positivo de HIV (2,6 por ciento). La mejor agudeza visual corregida de 0,3 o menos se mostró en el 100 por ciento de ellos antes del tratamiento, y después de este los 39 tenían entre 0,8 y 1,0. En el 100 por ciento de los casos se observaron cuerpos de inclusión basofílicos en el endotelio corneal, que fueron apreciables mediante la microscopia confocal. Conclusiones: Existe una relación entre la presencia de cuerpos de inclusión basofílicos en el endotelio corneal y las pruebas virológicas a citomegalovirus positivas, lo que puede permitir hacer PCR a casos que ya tienen confirmación mediante microscopia confocal de las características patognomónicas del endotelio corneal(AU)

Objective: Describe the characteristics of the corneal endothelium by confocal microscopy in patients with inclusion basophilic lesions as pathognomonic signs of cytomegalovirus endotheliitis. Methods: An observational study was conducted of clinical cases of patients diagnosed with cytomegalovirus endotheliitis attending the Cornea Service at Ramón Pando Ferrer Cuban Institute of Ophthalmology from February 2010 to June 2018. The study sample was 39 patients with a clinical diagnosis of endotheliitis who underwent confocal microscopy with a Confoscan 4 device (Nidek Technologies). Results: Of the 39 patients examined, 29 were male and 10 were female. All were aged 41-60 years. 97.4 percent had a history of viral conjunctivitis and only one was HIV positive (2.6 percent). Best corrected visual acuity was 0.3 or less in 100 percent before treatment, and 0.8 to 1.0 after treatment. Inclusion basophilic bodies visible by confocal microscopy were observed in the corneal endothelium of all patients. Conclusions: A relationship exists between the presence of inclusion basophilic bodies in the corneal endothelium and virological tests positive for cytomegalovirus, making it possible to perform PCR testing in cases with confocal microscopy confirmation of the pathognomonic characteristics of the corneal endothelium(AU)

In vivo confocal microscopy findings after SMILE refractive surgery technique.

OBJECTIVE: To evaluate confocal microscopy finding after SMILE surgery by in vivo confocal microscopy and stromal lenticule and stromal interface accuracy planed vs measured. METHOD: Thirty eyes of 15 patients were evaluated before and 1 month after SMILE surgery by using confocal microscopy. Cellular morphology was studied. Planed stromal lenticule thickness was compared vs measured stromal lenticule thickness after surgery by comparing the total stromal corneal thickness before vs after surgery. Stromal corneal interface was evaluated and depth of this planed interface was compared vs measured post-surgery interface depth. RESULTS: Sub-epithelial nerve plexus was absent at 1 month after surgery. Activated keratocytes were found before and after stromal corneal interface depth. There was not statistical significant difference between mean planed stromal lenticule vs. post-surgery measured (102.3 ± 25.7 vs. 104.2 ± 29.9 mm; p = 0.73). There was not statistical significant difference between mean planed stromal interface depth vs. post-surgical measured (113.7 ± 8.2 vs. 120.5 ± 17.3 mm; p = 0.058). CONCLUSIONS: Confocal microscopy is useful to evaluate changes after SMILE surgery; lenticule thickness and stromal interface depth are exact.

OBJETIVO: Evaluar los hallazgos mediante microscopía confocal in vivo de pacientes operados de cirugía SMILE, la exactitud del lentículo estromal y la profundidad de la interfaz estromal planeados versus medidos. MÉTODO: Treinta ojos de 15 pacientes operados de SMILE se estudiaron mediante microscopía confocal antes y al mes de la cirugía. Se evaluaron la morfología celular, el espesor del lentículo estromal planeado versus el medido en el posoperatorio mediante la diferencia entre el espesor estromal preoperatorio y el postoperatorio, así como la interfaz estromal, y se comparó la profundidad de esta interfaz estromal planeada con la medida en el posoperatorio. RESULTADOS: Se observó la ausencia del plexo nervioso subepitelial al mes del posoperatorio y una activación de queratocitos anterior y posterior a la profundidad de la interfaz estromal. No hubo diferencia en el lentículo estromal planeado versus el medido en el posoperatorio (102.3 ± 25.7 vs. 104.2 ± 29.9 mm; p = 0.73). No hubo diferencia entre la profundidad de la interfaz estromal planeada y medida (113.7 ± 8.2 vs. 120.5 ± 17.3 mm; p = 0.058). CONCLUSIONES: Tras la cirugía SMILE se pueden evaluar los cambios mediante microscopía confocal; el espesor del lentículo y la profundidad de la interfaz son exactos.

Liver structural injury in leptin-deficient (ob/ob) mice: lipogenesis, fibrogenesis, inflammation, and apoptosis / Lesión estructural del hígado en ratones con deficiencia de leptina (ob/ob): lipogénesis, fibrogénesis, inflamación y apoptosis

SUMMARY: 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 leptin- deficient 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.

RESUMEN: La enfermedad del hígado graso no alcohólico (HGNA) puede progresar de la esteatosis a esteatohepatitis no alcohólica (ENA), alcanzando un estado de cirrosis y posiblemente carcinoma hepatocelular. Se estudió el hígado de ratones C57BL / 6J de tres meses de edad (tipo salvaje, grupo WT, n = 10) y ratones obesos con deficiencia de leptina (grupo ob/ob, n = 10), centrándose en los mecanismos asociados con la activación de las células estrelladas hepáticas (HSC) y profibrogénesis. El hígado de los animales obesos ob/ob mostró esteatosis, aumento de la expresión génica de la lipogénesis, inflamación, aumento de la expresión génica proinflamatoria, infiltrado inflamatorio y posible apoptosis ligada a una alta expresión de Caspasa 3. En ratones ob/ob, las sec- ciones de hígado se marcaron en las zonas fibróticas con anti-alfa- actina de músculo liso (alfa-SMA) y anti-Reelin, pero no en los ratones WT. Además, la expresión del gen alfa-SMA fue mayor en el hígado del grupo ob/ob que en el grupo WT. Las expresiones génicas profibrogénicas fueron paralelas a la inmunofluorescencia anti-alfa-SMA y anti-Reelin, lo que sugiere la activación de las HSC. En los animales ob/ob, hubo un aumento de las expresiones génicas involucradas con la lipogénesis (receptor activado por proliferador de peroxisoma gamma, efector c similar a DFFA inductor de muerte celular, proteína de unión al elemento regulador de esterol-1c y sintasa de ácidos grasos), pro-fibrogénesis (factor de crecimiento transformante beta 1, proteínas Smad-3, proteína-1 asociada a Yes, receptor beta del factor de crecimiento derivado de plaquetas de proteínas), proinflamación (factor de necrosis tumoral alfa e interleucina-6) y apoptosis (caspasa 3). ). En conclusión, los resultados en animales obesos ob/ob proporcionan una pista de los eventos en humanos. Desde un punto de vista traslacional, el control de estos objetivos puede ayudar a mitigar los efectos hepáticos de la obesidad humana y la progresión de HGNA a ENA.

Imagenología diferencial por microscopia confocal in vivo entre la epiteliopatía por acanthamoeba y la queratitis por herpes simple / In vivo confocal microscopy imaging for the differential diagnosis of Acanthamoeba epitheliopathy and herpes simplex keratitis

La queratitis por Acanthamoeba es una infección corneal de baja incidencia, condicionada por varios factores, pero con manifestación clínica variada y sintomatología típica. En su fase epitelial debe ser diferenciada de otras queratitis, específicamente de la queratitis por herpes simple, por la similitud entre ambas en cuanto a las características de la lesión corneal. La microscopia confocal in vivo constituye una alternativa diagnóstica. Es una biopsia fotográfica en cuyas imágenes podemos describir los quistes y trofozoítos de Acanthamoeba desde etapas iniciales, que nos ayudan a diferenciarla de otros tipos de queratitis e iniciar el tratamiento precoz. Se realizó una búsqueda de artículos publicados, con el objetivo de mostrar las imágenes por microscopia confocal de la fase epitelial de la infección corneal por Acanthamoeba y herpes simple. Se utilizó la plataforma Infomed, específicamente la Biblioteca Virtual de Salud(AU)

Acanthamoeba keratitis is a low-incidence corneal infection caused by several factors and characterized by a variety of clinical manifestations and typical symptoms. In its epithelial phase, it should be differentiated from other keratitis, particularly from herpes simplex keratitis, due to the similar characteristics of the corneal lesion. In vivo confocal microscopy is a diagnostic alternative consisting in a photographic biopsy showing images of Acanthamoeba cysts and trophozoites since their initial stages, thus allowing differentiation from other types of keratitis and the initiation of early treatment. A search was conducted of published papers with the purpose of showing confocal microscopy images of the epithelial phase of Acanthamoeba and herpes simplex corneal infection. Use was made of the platform Infomed, specifically the Virtual Health Library(AU)

Adaptación marginal de coronas de disilicato de litio obtenidas mediante técnicas de escaneo (CAD/CAM): análisis in vitro con microscopía confocal / Marginal adaptation of lithium disilicate crowns obtained by scanning techniques (CAD/CAM): in vitro confocal microscopy analysis

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 des- pués 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 gru- pos, 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 Den- tal 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 mar- ginal vertical antes y después de la cristalización mostró una diferencia estadísticamente significativa (p=0,038). Conclusiones. Las coronas de disilicato de litio obtenidas me- diante escaneo directo (CAD/CAM) presentaron menor desajuste marginal vertical. La etapa de cristalización afectó la adaptación marginal de las coronas.

Objective. This study aimed to evaluate lithium disilicate marginal adaption on crowns by scanning techniques (CAD/CAM), before and after crystallization, through confocal microscopy (CM) in vitro analysis. Methods. Sixteen polyurethane replicas were per- formed from tooth 1.4, of a typodont model, prepared for a full crown. The replicas were divided into two groups, according to the scanning technique: Indirect Technique (Group IND, n=08), where dental stone models were scanned with a laboratory scanner (inEos X5, Sirona Dental Systems) and Direct Technique (Group DIR, n=08), where typodont models were scanned with an intraoral scanner (CEREC BlueCam, Sirona Dental Systems). Then, the lithium disilicate crowns (IPS e.max CAD, Ivoclar Vivadent) were milled (inLab MC XL, Sirona Dental Systems) and adapted to the replicas. Margin- al adaptation was evaluated with CM analysis before and after lithium disilicate crystalli- zation. Data were analyzed with the Mann-Whitney, t test, and Wilconxon test (α=0.05). Results. There was a statistically significant difference in horizontal marginal adaptation between IND and DIR groups after crystallization (p=0.05). In IND group, the compar- ison of vertical marginal adaptation before and after crystallization showed a statistically significant difference (p=0.038). Conclusions. Lithium disilicate crowns obtained by direct scanning technique (CAD/CAD) showed less vertical marginal maladjustment. The crystallization stage affected the crown's marginal adaptation.

Descubrimiento de fármacos basado en imagenología de células vivas / Drug discovery based on live cell imaging

RESUMEN El diseño eficiente de compuestos aprovechando las características estructurales de las moléculas y la búsqueda eficiente de dianas terapéuticas, ha proporcionado herramientas efectivas en la investigación de nuevos tratamientos cuando esta se enfoca en mecanismos celulares de la enfermedad. Los cambios fenotípicos producidos por la interacción in vitro entre molécula-diana, pueden controlarse cuantitativamente mediante imagenología de células vivas. Para garantizar una interacción adecuada, es necesario considerar diferentes elementos cruciales: 1. Las características estructurales y la dinámica molecular del compuesto a evaluar. 2. La relevancia del blanco para la fisiopatología de interés. Sin embargo, el desconocimiento del panorama general en el descubrimiento de fármacos, desde problemáticas estructurales y celulares, ha enlentecido la búsqueda de nuevos tratamientos. Esta revisión descriptiva de tema presenta algunos aspectos estructurales importantes para la caracterización de compuestos como candidatos terapéuticos, y aproximaciones experimentales para desarrollo de sistemas celulares. Los tópicos discutidos se enfocan en la monitorización por imagenología de células vivas y así mismo proporcionamos ejemplos relevantes. La monitorización de efectos fenotípicos producidos por interacciones entre candidato químico y blanco terapéutico en un sistema celular puede favorecer la búsqueda eficiente de moléculas potencialmente terapéuticas.

SUMMARY 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.

Microscopia confocal de la córnea en pacientes diabéticos / Confocal microscopy of the cornea in diabetic patients

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 por ciento en los pacientes con diabetes mellitus tipo 1; el sexo femenino en los pacientes con diabetes mellitus tipo 2 (60 por ciento) y aparentemente sanos (56,7 por ciento). En los pacientes con diabetes mellitus tipo 1 fueron más frecuentes las edades entre 45 y 54 años (33,3 por ciento) y entre 55 y 66 años en los pacientes con diabetes mellitus tipo 2 y aparentemente sanos con 60 y 40 por ciento respectivamente. La morfología del epitelio y el estroma corneal fue normal en el 86,7 y 87,3 por ciento respectivamente. Predominaron las alteraciones de la morfología endotelial en pacientes diabéticos tipo 1 (73,3 por ciento), así como el polimegatismo y el pleomorfismo (73,3 y 56,7 por ciento 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(AU)

ABSTRACT Objective: Identify the morphological characteristics of the corneal epithelium, stroma and endothelium, as well as the cell density of the endothelium by means of confocal microscopy of the cornea in diabetic patients. Methods: A descriptive comparative study was conducted of 90 eyes: 60 from diabetic patients (30 type 1 and 30 type 2) and 30 from supposedly healthy patients, at Ramón Pando Ferrer Cuban Institute of Ophthalmology from January 2012 to January 2017. Results: A predominance was found of the male sex (66.7 percent) among patients with diabetes mellitus type 1 and of the female sex among patients with diabetes mellitus type 2 (60 percent) and seemingly healthy patients (56.7 percent). The most common age ranges were 45-54 years for patients with diabetes mellitus type 1 (33.3 percent) and 55-66 years for patients with diabetes mellitus type 2 (60 percent) and seemingly healthy patients (40 percent). Morphology of the corneal epithelium and stroma was normal in 86.7 percent and 87.3 percent, respectively. In type 1 diabetic patients there was a predominance of endothelial morphological alterations (73.3 percent), polymegethism and pleomorphism (73.3 percent and 56.7 percent, respectively) and the lowest cell density (2 222.76 cells /mm2). Conclusions: Absence of morphological alterations of the corneal epithelium and stroma, as well as the presence of polymegethism and pleomorphism were the most common findings(AU)

Ex Vivo Confocal Microscopy Using Fusion Mode and Digital Staining: Changing Paradigms in Histological Diagnosis. / Microscopia confocal ex vivo con método de fusión y tinción digital: cambiando paradigmas en el diagnóstico histológico.

The ex vivo confocal microscope is an imaging system designed to analyze freshly excised tissue using two diode lasers with different wavelengths. The technique can dramatically reduce margin analysis times and offers a sensitivity of 88% and a specificity of 89% relative to histopathology. A new technology has recently been developed that produces images more quickly and with a higher resolution than before. By means of a fusion mode that combines simultaneously scanned fluorescence and reflectance images, it produces digitally stained images that simulate the effect of hematoxylin-eosin staining. Application of this new technology has opened the door to real-time tissue diagnostics.

Caracterización de la membrana basal, los plexos nerviosos y el grosor corneal mediante microscopia confocal en pacientes diabéticos / Characterization of basal membrane, nervous plexus and corneal thickness by confocal microscopy in diabetic patients

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 por ciento de los ojos de pacientes con diabetes mellitus tipo 1, y en el 10 por ciento 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 por ciento cada uno. En los diabéticos tipo 2, predominó un plexo nervioso (40 por ciento) y en pacientes sanos predominaron 4 plexos nerviosos (66,7 por ciento). Ambos grupos de pacientes diabéticos presentaron plexos verticales tortuosos (40 por ciento y 53,3 por ciento respectivamente) y plexos nerviosos oblicuos en el grupo de pacientes supuestamente sanos (80 por ciento). 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(AU)

ABSTRACT Objective: To identify the morphological and morphometric characteristics of the corneal layers by confocal microscopy in diabetic patients. Methods: A descriptive and comparative study was carried out in 90 eyes, 60 of which belonged to diabetic patients (30 type 1 and 30 type 2) and 30 to supposedly healthy patients. The study was conducted at Ramón Pando Ferrer Cuban Institute of Ophthalmology, between January 2012 and January 2017. Results: Epithelial thickness, stroma and corneal endothelium was greater in the eyes of type 1 diabetics with a mean of 54.6, 506.7 and 26.7 microns, respectively. The epithelial basement membrane was observed in 20 percent of the eyes of patients with type 1 diabetes mellitus, and in 10 percent of type 2 diabetics. The group of type 1 diabetics showed one and two nerve plexuses per confocal microscopy field for 33.3 percent each. In type 2 diabetics, one nerve plexus (40 percent) predominated, while in healthy patients, four nerve plexuses (66.7 percent) predominated. Both groups of diabetic patients presented tortuous vertical plexuses (40 percent and 53.3 percent, respectively) and oblique nerve plexuses in the group of supposedly healthy patients (80 percent). Conclusions: The study of the cornea by confocal microscopy in diabetic patients showed greater total and layered corneal thicknesses, visible basal membrane, decrease in the sub-basal nerve plexus with vertical arrangement, and tortuous fiber(AU)

Microscopia confocal no auxílio diagnóstico de Distrofia Corneana de Schnyder / In vivo confocal microscopy as a diagnostic tool in Schnyder Corneal Dystrophy's case

Resumo Neste relato, descrevemos um caso de Distrofia corneana de Schnyder que apresentou o desfecho de seu diagnóstico baseado em achados característicos na microscopia confocal, ferramenta que se aponta em destaque no universo oftalmológico.

Abstract 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).

The effect of panretinal photocoagulation on confocal laser scanning ophthalmoscopy and stereo photographic parameters of optic disk topography in patients with diabetic retinopathy / Efeito da panfotocoagulação retiniana na oftalmoscopia confocal de varredura a laser e parâmetros da estereofotografia do disco óptico em pacientes com retinopatia diabética

ABSTRACT Purpose: To determine the effect of panretinal photocoagulation on optic disk topographic parameters in non-glaucomatous patients with proliferative diabetic retinopathy. Methods: This was a prospective, single-center, observational study. Thirty-eight eyes of 26 patients with diabetes underwent panretinal photocoagulation for proliferative diabetic retinopathy. Stereoscopic disk photographs and optic nerve head parameters were evaluated using the Zeiss fundus camera and the confocal scanning laser ophthalmoscope (Heidelberg Retinal Tomograph), respectively, at baseline and 12 months after the completion of panretinal photocoagulation. Results: Thirty-eight eyes of 26 patients (15 female) with a mean age of 53.7 (range 26-74) years were recruited. No significant difference was found between the stereo photography determined mean horizontal and vertical cup-to-disk ratio before and after panretinal photocoagulation treatment (p=0.461 and 0.839, respectively). The global values of the optic nerve head parameters analyzed with the HRT3 showed no significant change from baseline to 12 months, including the disk area, cup area, rim area, cup volume, rim volume, cup-to-disk area ratio, linear cup-to-disk ratio, mean cup depth, maximum cup depth, cup shape measure, height variation contour, mean retinal nerve fiber layer thickness, and cross-sectional area. Conclusion: Our results suggest that panretinal photocoagulation does not cause morphological optic disk changes in patients with diabetic proliferative retinopathy after 1 year of follow-up.

RESUMO Objetivo: Determinar o efeito da panfotocoagulação retiniana nos parâmetros topográficos do disco óptico em pacientes não glaucomatosos com retinopatia diabética proliferativa. Métodos: Este é um estudo observacional pros­pectivo e unicêntrico. Trinta e oito olhos de 26 pacientes dia­béticos foram submetidos à panfotocoagulação retiniana para retinopatia diabética proliferativa. As estereofotografias e os parâmetros do disco óptico foram avaliados usando o retinógrafo Visucam da Zeiss e o oftalmoscópio confocal de varredura a laser (Heidelberg Retinal Tomograph), respectivamente, no início e 12 meses após a conclusão da panfotocoagulação. Resultados: Trinta e oito olhos de 26 pacientes (15 mulheres) com média de idade de 53,7 anos (intervalo de 26-74) foram recrutados. Nenhuma diferença significativa foi encontrada entre a média horizontal e vertical para relação escavação/disco óptico determinadas pelas estereo­fotografias antes e após o tratamento com panfotocoagulação retiniana (p=0,461 e 0,839, respectivamente). Os valores globais dos parâmetros do disco óptico analisados com a tomografia de varredura a laser não mostraram nenhuma mudança significativa entre o início até os 12 meses, incluindo disk area, cup area, rim area, cup volume, rim volume, C/D area ratio, linear C/D ratio, mean cup depth, maximum cup depth, cup shape measure, height variation contour, mean retinal nerve fiber layer thickness e cross-sectional area. Conclusão: Nossos resultados sugerem que a panfotocoagulação retiniana não causa alterações morfológicas no disco óptico em pacientes com retinopatia diabética proliferativa após um ano de seguimento.

In Vivo Reflectance Confocal Microscopy: Emerging Role in Noninvasive Diagnosis and Monitoring of Eczematous Dermatoses. / Microscopia confocal de reflectancia in vivo: papel emergente en el diagnóstico no invasivo, así como en el seguimiento de las dermatosis eccematosas.

Dermatologic diagnosis and monitoring have been dependent largely on visual grading. A skin biopsy is performed in case of diagnostic uncertainty, but can be traumatic, and results are delayed due to time for specimen transport and processing. Biopsies also destroy specimens, prohibiting lesion evolution monitoring. In vivo reflectance confocal microscopy (RCM) offers a diagnostic alternative to skin biopsy. RCM captures real-time, high-resolution images, and has been piloted for the evaluation of various dermatologic conditions. Identification of unique RCM features may distinguish dermatoses with similar clinical morphologies. Allergic contact dermatitis (ACD) and irritant contact dermatitis (ICD) are diagnosed by patch testing that currently uses a subjective scoring system. RCM has increasingly been studied for early detection and severity grading of CD. Common RCM features shared by ACD and ICD are stratum corneum disruption, vesicle formation, exocytosis, spongiosis, and parakeratosis. Features unique to ACD are vasodilation, increased epidermal thickness, intercellular edema, and acanthosis. Features unique to ICD are detached corneocytes and targetoid keratinocytes. This review summarizes the use of RCM in evaluating contact eccematous conditions and aims to spark future research and interest in this promising tool.