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.

Reflectance Confocal Microscopy for Inflammatory Skin Diseases.

In vivo reflectance confocal microscopy (RCM) is a relatively novel non-invasive tool for microscopic evaluation of the skin used prevalently for diagnosis and management of skin tumour. Its axial resolution, its non-invasive and easy clinical application represents the goals for a large diffusion of this technique. During the last 15 years, RCM has been demonstrated to be able to increase the sensibility and sensitivity of dermoscopy in the diagnosis of skin tumours integrating in real time clinic, dermoscopic and microscopic information useful for the definition of malignancy. Despite to date, no large comparative studies on inflammatory skin diseases has been published in the literature, several papers already showed that RCM has a potential for the evaluation of the descriptive features of the most common inflammatory skin diseases as psoriasis, lupus erythematosus, contact dermatitis and others. The aim of the application of this technique in non-neoplastic skin diseases has been prevalently focused on the possibility of clinical diagnosis confirmation, as well as therapeutic management. Moreover, the use of RCM as driver for an optimised skin biopsy has been also followed in order to reduce the number of unsuccessful histopathological examination. In this review article we describe the confocal features of the major groups of inflammatory skin disorders focusing on psoriasiform dermatitis, interface dermatitis and spongiotic dermatitis.

Usefulness of confocal microscopy in distinguishing between basal cell carcinoma and intradermal melanocytic nevus on the face.

The clinical distinction between basal cell carcinoma (BCC) and intradermal melanocytic nevus lesions on the face can be difficult, particularly in young patients or patients with multiple nevi. Dermoscopy is a useful tool for analyzing characteristic dermoscopic features of BCC, such as cartwheel structures, maple leaf-like areas, blue-gray nests and dots, and ulceration. It also reveals arborizing telangiectatic vessels and prominent curved vessels, which are typical of BCC, and comma vessels, which are typical of intradermal melanocytic nevi. It is, however, not always easy to distinguish between these 2 conditions, even when dermoscopy is used. We describe 2 facial lesions that posed a clinical and dermoscopic challenge in two 38-year-old patients; confocal microscopy showed separation between tumor nests and stroma and polarized nuclei, which are confocal microscopy features of basal cell carcinoma.

Clinical usefulness of reflectance confocal microscopy in the management of facial lentigo maligna melanoma.

Facial lentigo maligna melanoma can be a diagnostic challenge in daily clinical practice as it has similar clinical and morphological features to other lesions such as solar lentigines and pigmented actinic keratoses. Confocal microscopy is a noninvasive technique that provides real-time images of the epidermis and superficial dermis with cellular-level resolution. We describe 3 cases of suspected facial lentigo maligna that were assessed using dermoscopy and confocal microscopy before histopathology study. In the first case, diagnosed as lentigo maligna melanoma, presurgical mapping by confocal microscopy was performed to define the margins more accurately. In the second and third cases, with a clinical and dermoscopic suspicion of lentigo maligna melanoma, confocal microscopy was used to identify the optimal site for biopsy.

In vivo confocal microscopy findings of cornea and tongue mucosa in patients with Sjögren's syndrome.

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.

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.

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.

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.

Ocular surface analysis in patients diagnosed with X-linked ichthyosis. / Análisis de superficie ocular en pacientes diagnosticados de ictiosis X.

Seven patients (14 eyes) diagnosed with X-linked ichthyosis were studied using the Schirmer test, biomicroscopy, tonometry, endothelial count, optical coherence tomography, Pentacam®, ocular surface analyser, and confocal microscopy. The mean age was 33.83±20.17 years (range: 7-64 years). The most frequent findings in biomicroscopy were Meibomian glands dysfunction (83.3%) and stromal corneal opacities (33%). The tear break-up time was found shortened in 25% of the eyes. Confocal microscopy (both eyes) revealed activated keratocytes with hyper-reflective particles inside them in the anterior stroma and outside them in the posterior stroma. It is believed that the inclusion of the use of confocal microscopy will help in a better understanding of the corneal pathology associated with ichthyosis X, as well as new characteristics of these patients.

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.

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.

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.

Reflectance Confocal Microscopy: A Promising Tool to Identify Malignancy in Melanocytic Lesions Exhibiting a Dermoscopic Island. / Microscopia confocal de reflectancia: una herramienta útil para diagnosticar malignidad en lesiones melanocíticas que presenten el signo de la isla.

The dermoscopic island is described as a well-defined area in a melanocytic lesion, with a different dermoscopic pattern from the rest of the lesion. It is predictive of melanoma, particularly when the pattern of the island is atypical. We present the reflectance confocal microscopy (RCM) findings in 3 lesions with dermoscopic islands: nevus-associated melanoma, melanocytic nevus, and in situ melanoma. The nevus-associated melanoma and in situ melanoma presented cellular atypia (atypical cells in isolation or forming nests) and architectural distortion on RCM. The nevus presented a dermoscopic island with a typical globular pattern with dense nests and no atypia on RCM. Dermoscopic island is mainly associated with in situ and nevus-associated melanomas. RCM offers good cellular resolution to the depth of the reticular dermis and is useful for diagnosing of melanomas presenting a dermoscopic island.

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)

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.