Histological and Ultrastructural Changes in Lungs One Year Post Asymptomatic or Mild SARS-CoV-2 Infection: Evaluating the Potential for Active Virus Presence / Cambios Histológicos y Ultraestructurales en los Pulmones un Año Después de una Infección Asintomática o Leve por SARS-CoV-2: Evaluación del Potencial de Presencia Activa del Virus

SUMMARY: Prior research on post-COVID-19 or long COVID primarily focused on the presence of SARS-CoV-2 mostly in symptomatic patients. This study aimed to investigate the persistence of SARS-CoV-2 after 1 year of asymptomatic or mild COVID-19. SARS-CoV-2 infected and control K18-hACE2 transgenic mice (n=25) were studied. Moderate and severe symptomatic subjects were sacrificed after eight days, while mild or asymptomatic mice were kept in BSL-III for twelve months. Analyses included general condition, histochemistry, immunohistochemistry, transmission electron microscopy, and qRT-PCR. Lungs from the twelve-month group showed thickening of alveolar walls, with some lungs exhibiting the recruitment of inflammatory cells, the presence of SARS- CoV-2 mRNA, immunopositivity for the SARS-CoV-2 spike protein, and TEM showed viruses (60-125 nm) within vesicles, indicating continued replication. Certain lung samples showed persistent SARS-CoV-2 presence in Club cells, endothelial cells, and macrophages. The eight-day group exhibited viral interstitial pneumonitis, SARS-CoV-2 immunopositivity, and mRNA. The eight-day hearts displayed viral mRNA, while the twelve-month hearts tested negative. Some asymptomatic twelve-month subjects presented reduced surfactant, basal membrane thickening, fibrosis, and mild autonomic nerve degeneration. In this study conducted on mice, findings indicate the potential for chronic persistence of SARS-CoV-2 in the lungs one year post initial mild or asymptomatic infection, which could suggest the possibility of recurrent episodes in similar human conditions. The observed thickening of alveolar walls and potential fibrotic areas in these mice may imply an increased risk of post-COVID fibrosis in humans. Furthermore, the presence of SARS-CoV-2-positive inflammatory cells in some asymptomatic murine cases could herald a progression toward ongoing inflammation and chronic lung disease in humans. Therefore, the necessity for further studies in human subjects and vigilant monitoring of high-risk human populations is underscored.

Investigaciones anteriores sobre COVID-19 o COVID prolongado se centraron principalmente en la presencia de SARS-CoV-2 principalmente en pacientes sintomáticos. Este estudio tuvo como objetivo investigar la persistencia del SARS-CoV-2 después de 1 año de COVID-19 asintomático o leve. Se estudiaron ratones transgénicos K18-hACE2 infectados con SARS-CoV-2 y de control (n=25). Los animales con síntomas moderados y graves se sacrificaron después de ocho días, mientras que los ratones con síntomas leves o asintomáticos se mantuvieron en BSL-III durante doce meses. Los análisis incluyeron estado general, histoquímica, inmunohistoquímica, microscopía electrónica de transmisión y qRT- PCR. Los pulmones del grupo de doce meses mostraron engrosamiento de las paredes alveolares, y algunos pulmones exhibieron reclutamiento de células inflamatorias, presencia de ARNm del SARS-CoV-2, inmunopositividad para la proteína de la espícula del SARS-CoV-2 y TEM mostró virus (60 -125 nm) dentro de las vesículas, lo que indica una replicación continua. Ciertas muestras de pulmón mostraron una presencia persistente de SARS- CoV-2 en exocrinocitos bronquiolares, células endoteliales y macrófagos. El grupo de ocho días presentó neumonitis intersticial viral, inmunopositividad al SARS-CoV-2 y ARNm. Los corazones de ocho días mostraron ARNm viral, mientras que los corazones de doce meses dieron negativo. Algunos animales asintomáticos de doce meses presentaron disminución del surfactante, engrosamiento de la membrana basal, fibrosis y degeneración leve del nervio autónomo. En este estudio realizado en ratones, los hallazgos indican la posibilidad de persistencia crónica del SARS-CoV-2 en los pulmones un año después de la infección inicial leve o asintomática, lo que podría sugerir la posibilidad de episodios recurrentes en condiciones humanas similares. El engrosamiento observado de las paredes alveolares y las posibles áreas fibróticas en estos ratones puede implicar un mayor riesgo de fibrosis post-COVID en humanos. Además, la presencia de células inflamatorias positivas para SARS- CoV-2 en algunos casos murinos asintomáticos podría presagiar una progresión hacia una inflamación continua y una enfermedad pulmonar crónica en humanos. Por lo tanto, se subraya la necesidad de realizar más estudios en seres humanos y realizar un seguimiento atento de las poblaciones humanas de alto riesgo.

The association of bacterial biofilm and middle ear mucosa in patients with mucosal chronic suppurative otitis media.

OBJECTIVES: To evaluate the bacterial biofilm's role in mucosal chronic suppurative otitis media (CSOM) utilizing scanning electron microscopy (SEM). METHODS: This study involved 123 participating patients with active and inactive mucosal CSOM who underwent tympanomastoid surgery. SEM was used to examine middle ear mucosa biopsies for the development of biofilms. Middle ear discharge or mucosal swabs from patients were cultured to detect any bacterial growth. The biofilm formation was correlated to the culture results. RESULTS: The biofilm was present in 69.9 % of patients (59% of them were with active mucosal CSOM) and absent in 30.1% of the patients (70% of them were with inactive mucosal CSOM), being more statistically significant in active mucosal CSOM (p-value = 0.003). A correlation that was statistically significant was found between active mucosal CSOM and higher grades (3 and 4) of biofilms (p-value <0.05). The mucosal CSOM type and the results of the culture had a relationship that was statistically significant (p-value <0.001). 60% of patients had positive culture (70% of them were with active mucosal CSOM). There was a statistically significant relation between Pseudomonas aeruginosa bacterial growth and active mucosal CSOM (p-value = 0.004) as well as higher grades of biofilms in mucosal CSOM. CONCLUSION: Mucosal CSOM, especially the active type, is a biofilm-related disease. There is a significant relation between the state of mucosal CSOM (active or inactive) and culture results with predominance of Pseudomonas aeruginosa bacterial growth in active mucosal CSOM and in higher grades of biofilms in mucosal CSOM.

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.

Alpaca (Vicugna pacos) Skin Analyzed Under Architecture and Structure / Piel de Alpaca (Vicugna pacos) Analizada Bajo Arquitectura y Estructura

SUMMARY: This study evaluated the morphology of alpacas skin. Biopsies were collected and samples were fixed in 10 % neutral buffered formalin for histological procedures. The sections were stained with hematoxylin and eosin, picrosirius red and Masson's trichrome. Types I, III and IV collagen were analyzed by immunohistochemistry. The derma presented sebaceous and sweat glands, as well as follicular groups with medullated fibers. Type I and type IV collagen were observed at epidermis and dermis as well as in glandular structures and hair follicles. The collagen III, was observed only in dermis.

Este estudio evaluó la morfología de la piel de alpacas. Se recogieron biopsias y las muestras se fijaron en formalina tamponada neutra al 10 % para procedimientos histológicos. Las secciones se tiñeron con hematoxilina y eosina, rojo picrosirius y tricrómico de Masson. El colágeno tipo I, III y IV se analizó mediante inmunohistoquímica. La dermis presentó glándulas sebáceas y sudoríparas, así como grupos foliculares con fibras medulares. Se observó colágeno tipo I y tipo IV en la epidermis y la dermis, así como en estructuras glandulares y folículos pilosos. El colágeno III, se observó únicamente en la dermis.

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

La terminología usada para describir los diferentes hallazgos en la microscopía confocal de reflectancia (MCR), tanto en lesiones melanocíticas, como en no melanocíticas se ha consensuado en inglés. En el presente trabajo, se proponen los términos en español que mejor interpretan estos conceptos ya descritos para la MCR, mediante el consenso de expertos de distintas nacionalidades de habla hispana y utilizando el método DELPHI para el acuerdo final. Se obtuvieron 52 términos en total, de los cuales 28 fueron para lesiones melanocíticas y 24 para lesiones no melanocíticas. El uso de la nomenclatura propuesta permitirá una homogeneización y mejor entendimiento de las estructuras; una descripción más estandarizada en los registros clínicos y una mejor interpretación de estos informes por otros dermatólogos.(AU)

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.(AU)

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

La terminología usada para describir los diferentes hallazgos en la microscopía confocal de reflectancia (MCR), tanto en lesiones melanocíticas, como en no melanocíticas se ha consensuado en inglés. En el presente trabajo, se proponen los términos en español que mejor interpretan estos conceptos ya descritos para la MCR, mediante el consenso de expertos de distintas nacionalidades de habla hispana y utilizando el método DELPHI para el acuerdo final. Se obtuvieron 52 términos en total, de los cuales 28 fueron para lesiones melanocíticas y 24 para lesiones no melanocíticas. El uso de la nomenclatura propuesta permitirá una homogeneización y mejor entendimiento de las estructuras; una descripción más estandarizada en los registros clínicos y una mejor interpretación de estos informes por otros dermatólogos.(AU)

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.(AU)

Ganancia auditiva en pacientes intervenidos de cirugía del estribo por otosclerosis / Hearing gain in patients undergoing stapes surgery for otosclerosis

Introducción y objetivo: La otosclerosis es una causa de hipoacusia en jóvenes, con mayor frecuencia en mujeres. La cirugía del estribo es un procedimiento correctivo ampliamente aceptado, con el advenimiento de la tecnología y cambios en la técnica, surge la interrogante de si existen diferencias entre ellas. Objetivo: Evaluar si existen diferencias en la ganancia auditiva entre técnicas y abordajes de las cirugías del estribo en pacientes con otosclerosis Método: Se recabaron variables demográficas, clínicas y quirúrgicas. Se aplicó estadística descriptiva. Se empleó prueba U de Mann-Whitney para variables numéricas, así como Kruskal Wallis para comparación diferencias en tres o más grupos. Se consideró significativo un valor de p ≤ a 0.05. Resultados: Entre los años 2020 y 2023 se realizaron 55 cirugías de estribo por otosclerosis, de las cuales 20 se tuvieron que excluir. De 35 cirugías en 31 pacientes, la media de edad de 41.16 ± 8.64 años, 77.4% fueron mujeres, el 51.4 % fueron en el oído derecho; se presentaron comorbilidades en el 25.7%, las complicaciones 5 presentaron hipoacusia, el 88.6 % de los procedimientos se encontró un cierre satisfactorio de la brecha aérea y ósea. No se presentaron diferencia entre las técnicas de la cirugía de estribo y resultados audiológicos postquirúrgicos p=0.872, ni con el tipo de abordaje de visualización p=0.636. Discusión: Nuestros resultados son similares a lo que encontraron algunos autores, no obstante, aún sigue existiendo incertidumbre sobre la mejor técnica. Conclusiones: No se encontraron diferencias estadísticamente significativas en cuanto a la ganancia auditiva con el abordaje de visualización y el tipo de procedimiento en el estribo para la colocación de la prótesis. (AU)

Introduction and objective: Otosclerosis is a cause of hearing loss in young people, more frequently in women. Stapes surgery is a widely accepted corrective procedure, with the advent of technology and changes in technique, the question arises as to whether there are differences between them. Objective: To evaluate whether there are differences in hearing gain between techniques and approaches of stapes surgeries in patients with otosclerosis. Method: Demographic, clinical and surgical variables were collected. Descriptive statistics were applied. The Mann-Whitney U test was used for numerical variables, as well as the Kruskal Wallis test to compare differences in three or more groups. A p value ≤ 0.05 was considered significant. Results: Between 2020 and 2023, 55 stapes surgeries were performed for otosclerosis, of which 20 had to be excluded. Of 35 surgeries in 31 patients, mean age 41.16 ± 8.64 years, 77.4% were women, 51.4% were in the right ear; Comorbidities were present in 25.7%, 5 complications presented hearing loss, in 88.6% of the procedures a satisfactory closure of the air-bone gap was found. There was no difference between stapes surgery techniques and postsurgical audiological results p=0.872, nor with the type of visualization approach p=0.636. Discussion: Our results are similar to what some authors found, however, there is still uncertainty about the best technique. Conclusions: No statistically significant differences were found in terms of hearing gain with the visualization approach and the type of procedure in the stapes for placement of the prosthesis. (AU)

Hallazgos de microscopia confocal in vivo de la mucosa de la córnea y la lengua en pacientes con síndrome de Sjögren / In vivo confocal microscopy findings of cornea and tongue mucosa in patients with Sjögren's syndrome

Introducción y objetivos: Nuestro objetivo fue investigar si el síndrome de Sjögren (SS) tenía hallazgos distintivos en la microscopia confocal de la lengua de una manera no invasiva. Materiales y métodos: Este estudio retrospectivo de casos y controles evaluó los hallazgos de la microscopia confocal de córnea y lengua de los ojos derechos de 25 pacientes con ojo seco con deficiencia acuosa y 12 voluntarios sanos sin hallazgos de ojo seco. Hubo un total de 14 pacientes diagnosticados con ojo seco asociado a SS (SSDE), mientras que 11 casos fueron evaluados como ojo seco no Sjögren (NSDE). Resultados: Se observó una diferencia significativa en el recuento de células dendríticas a nivel del nervio subbasal corneal entre los grupos SSDE y NSDE (p=0,018). En el grupo SSDE, las imágenes de microscopia confocal de células inflamatorias dendritiformes hiperreflectantes en la mucosa de la lengua estaban a favor de la inflamación. Sin embargo, estos hallazgos no se encontraron en pacientes con NSDE o en controles. Conclusiones: Este estudio mostró que la microscopia confocal proporcionó una evaluación no invasiva de las células inflamatorias en la lengua de los pacientes con SS.(AU)

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.(AU)

Utilização da Microscopia Eletrônica de Varredura na detecção de matérias estranhas em águas envasadas / Scanning Electron Microscopy as a tool to detect and quantify microscopic physical hazards in bottled mineral waters

Este trabalho objetiva averiguar a presença de matérias estranhas presentes em águas minerais envasadas e sugerir a utilização da Microscopia Eletrônica de Varredura (MEV), como técnica adequada para detecção e contagem. Oito amostras de água mineral foram obtidas e subamostras de 100 mL foram filtradas em membranas, que foram secas e recobertas com platina e observadas em MEV. Áreas de 21.025 µm2 foram focalizadas e as partículas contadas. Realizaram-se análise de variância e o teste de Regwq para a comparação das médias das contagens. Diferentes partículas visualizadas foram registradas sob distintas magnificações. A análise de variância revelou alta significância entre a quantidade de matérias estranhas das amostras, indicando que as amostras diferiram bastante quanto a essa característica. As médias obtidas obedeceram a uma estratificação, mas foram estimadas com grande precisão, sugerindo-se que a subdivisão em 10 grupos de contagem e a extrapolação para o volume de 100 mL é uma estimativa confiável para a contagem de partículas filtradas nesse volume de água. Foi mostrado que o MEV se apresentou como instrumento auxiliar adequado para a contagem de matérias estranhas em amostras de águas minerais envasadas, especialmente quando suas dimensões não permitem sua identificação convencionalmente. (AU)

This work aims to investigate the presence of microscopic foreign materials in bottled mineral waters and suggest the use of Scanning Electron Microscopy (SEM) as a suitable technique for counting foreign matter. Eight samples of mineral water were obtained, and 100 mL subsamples were filtered through membranes, which were then dried, coated with platinum, and observed under SEM. Areas of 21,025 µm2 were analysed, and the particles present were counted. Variance analysis was carried out, and the Regwq test was applied to compare the counting averages. Samples of the dirt particles were imaged at different magnifications. The analysis of variance revealed high significance among the samples, indicating substantial differences between them. The averages followed a stratification pattern and were estimated with great precision, suggesting that the subdivision into 10 count groups and the extrapolation to 100 mL volume is a reliable estimate for the count of particles filtered in this volume of water. It was shown that SEM was a suitable instrument for counting and observing foreign matter and dirt in samples of bottled natural mineral waters, especially when their dimensions are too small to be detected by conventional methods. (AU)

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.

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.

Caracterização histopatológica de reações adversas a materiais de preenchimento estético em região oral e maxilofacial: estudo transversal multicêntrico / Histopathological characterization of adverse reactions to aesthetic filling materials in the oral and maxillofacial region: a multicenter cross-sectional study

O uso de materiais injetáveis estéticos na região orofacial vem aumentando exponencialmente, assim como as complicações associadas a estes procedimentos. As reações adversas a materiais estéticos podem se confundir com processos neoplásicos, tanto do ponto de vista clínico quanto microscópico. Este trabalho tem como objetivo principal descrever detalhadamente as características clínicas, demográficas, histopatológicas, histoquímicas e imuno-histoquímicas de reações adversas a diferentes preenchedores estéticos na região oral e maxilofacial. Foi realizado um estudo transversal multicêntrico, no qual as amostras foram coletadas de cinco laboratórios de patologia oral no Brasil e no México. Foram realizadas colorações de hematoxilina-eosina, Alcian blue, Sirius red e azul de toluidina, bem como imuno-histoquímica para CD68, CD3 e CD20. H&E também foi avaliado sob luz polarizada. Estatísticas descritivas foram realizadas. Vinte e três casos foram incluídos. Polimetilmetacrilato foi o material mais comum (n=10), seguido por silicone (n=4), ácido hialurônico (n=3), hidroxiapatita de cálcio (n=3), hidrogel de poliacrilamida (n=2) e ácido poli-L-láctico (n=1). Os pacientes foram principalmente mulheres (91,3%), com idade média de 50,65 anos. A maioria das reações afetou os lábios e foi assintomática, com tempo de evolução variável (7 dias a 10 anos), apresentando-se como nódulos de 58,07 mm em média. Polimetilmetacrilato e silicone apresentaram imagens negativas de formato arredondado, enquanto ácido hialurônico e hidrogel de poliacrilamida apresentaram-se como "piscinas" amorfas. A hidroxiapatita de cálcio apresentou estruturas poliédricas e o ácido poli-L-láctico apresentou formatos semelhantes a fendas. Ambos birrefringentes sob luz polarizada. Células gigantes foram comumente encontradas, exceto em silicone e ácido hialurônico. Granuloma de corpo estranho foi frequente em polimetilmetacrilato. Ácido hialurônico e hidrogel de poliacrilamida apresentaram metacromasia por azul de toluidina. Alcian blue foi positivo em todos os casos de ácido hialurônico. Mastócitos foram detectados em todos os materiais, exceto ácido hialurônico e hidrogel de poliacrilamida. Eosinófilos foram mais raros que mastócitos. Numerosas células CD68 positivas foram vistas em todos os casos. Todos os casos apresentaram células CD3 positivas, com quantidades variáveis. CD20 foi escasso ou negativo na maioria dos casos. Além do artigo científico, foi produzido um e-book de histopatologia, enfatizando o diagnóstico diferencial histológico destas lesões. Concluímos que apesar das semelhanças, há características específicas de cada material e da resposta do hospedeiro que auxiliam no diagnóstico histopatológico correto. Formato, tamanho e coloração do material no H&E são características-chave no diagnóstico diferencial. Uma reação intensa de macrófagos é observada em todos os preenchedores estéticos, frequentemente associada à formação de células gigantes. A imuno-histoquímica para CD68 e a coloração por azul de toluidina são as mais abrangentes para auxiliar no correto diagnóstico, sendo que outros marcadores podem ser úteis em casos específicos. Lesões neoplásicas também devem ser consideradas no diagnóstico diferencial histopatológico.

The use of aesthetic injectable materials in the orofacial region has been increasing exponentially, which has also led to an increase in complications associated with these procedures. Adverse reactions to aesthetic materials can be confused with neoplastic processes, both from a clinical and microscopic perspective. The main objective of this study is to describe in detail the clinical, demographic, histopathological, histochemical, and immunohistochemical characteristics of adverse reactions to different aesthetic fillers in the oral and maxillofacial region. A multicenter cross-sectional study was conducted, in which samples were collected from five oral pathology laboratories in Brazil and Mexico. Hematoxylin-eosin, Alcian blue, Sirius red, and toluidine blue staining, as well as immunohistochemistry for CD68, CD3, and CD20, were performed. H&E was also evaluated under polarized light. Descriptive statistics were conducted. Twenty-three cases were included. Polymethylmethacrylate was the most common material (n=10), followed by silicone (n=4), hyaluronic acid (n=3), calcium hydroxyapatite (n=3), polyacrylamide hydrogel (n=2), and poly-L-lactic acid (n=1). The patients were predominantly women (91.3%), with an average age of 50.65 years. Most reactions affected the lips and were asymptomatic, with variable evolution times (7 days to 10 years), presenting as nodules with an average size of 58.07 mm. Polymethylmethacrylate and silicone showed negative images with a rounded shape, while hyaluronic acid and polyacrylamide hydrogel presented as amorphous "pools." Calcium hydroxyapatite showed polyhedral structures, and poly-L-lactic acid exhibited fissure-like shapes. Both were birefringent under polarized light. Giant cells were commonly found, except in silicone and hyaluronic acid. Foreign body granuloma was frequent in polymethylmethacrylate. Hyaluronic acid and polyacrylamide hydrogel showed metachromasia with toluidine blue. Alcian blue was positive in all hyaluronic acid cases. Mast cells were detected in all materials except hyaluronic acid and polyacrylamide hydrogel. Eosinophils were rarer than mast cells. Numerous CD68- positive cells were seen in all cases. All cases presented CD3-positive cells, in varying amounts. CD20 was scarce or negative in most cases. In addition to the scientific article, a histopathology e-book was produced, emphasizing the histological differential diagnosis of these lesions. We conclude that despite similarities, there are specific characteristics of each material and host response that aid in the correct histopathological diagnosis. The shape, size, and staining of the material in H&E are key features in the differential diagnosis. An intense macrophage reaction is observed in all aesthetic fillers, often associated with the formation of giant cells. Immunohistochemistry for CD68 and toluidine blue staining are the most comprehensive for assisting in the correct diagnosis, although other markers may be useful in specific cases. Neoplastic lesions should also be considered in the histopathological differential diagnosis.

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.

A microscopia eletrônica de varredura pode ser usada para medir quantitativamente a rugosidade da dentina? / Can Scanning Electron Microscopy be used to quantitatively measure dentine roughness?

Objective: The aim of this study was to determine if SEM used with imaging software could be used to quantitatively determine the extent of dentine roughness due to tooth-brushing. Material and methods: Flat, polished dentine surfaces were subjected to 25,000 cycles of simulated tooth-brushing at 2 strokes per second with a load of 200 g. At the end of the brushing cycle, dentine surfaces were first assessed using profilometry and then subjected to SEM analysis. Ra (average roughness) readings were obtained for profilometric assessment. Using imaging software, the horizontal distance between adjacent characteristic grooves noted on micrographs was measured, ensuring that the middle of each crest of the associated groove was used as the reference point, the Crest-to-Crest distance (C-C distance). These two parameters were examined statistically for correlation. Results: When Spearman Rank tests were utilized the correlation between average Ra and crest to crest measurements was 0.709 (p <0.01). The Bland Altman plot, however, showed poor agreement between the two test parameters. Conclusion: Further work is needed to validate the use of this methodology in the quantitative assessment of tooth surface loss due to abrasion (AU)

Objetivo: O objetivo deste estudo foi determinar se o MEV usado com software de imagem poderia ser usado para determinar quantitativamente a extensão da rugosidade da dentina devido à escovação dentária. Material e Métodos: Superfícies dentinárias planas e polidas foram submetidas a 25.000 ciclos de escovação dentária simulada a 2 movimentos por segundo com uma carga de 200g. No final do ciclo de escovação, as superfícies dentinárias foram primeiro avaliadas por perfilometria e depois submetidas à análise MEV. Leituras de Ra (rugosidade média) foram obtidas para avaliação perfilométrica. Utilizando um software de imagem, foi medida a distância horizontal entre sulcos característicos adjacentes observados nas micrografias, garantindo que o meio de cada crista do sulco associado fosse usado como ponto de referência, a distância crista a crista (distância CC). Esses dois parâmetros foram examinados estatisticamente para correlação. Resultados: Quando foram utilizados os testes de Spearman Rank, a correlação entre a média de Ra e as medidas de crista a crista foi de 0,709 (p<0,01). O gráfico de Bland Altman, no entanto, mostrou fraca concordância entre os dois parâmetros de teste. Conclusão: Mais trabalhos são necessários para validar o uso desta metodologia na avaliação quantitativa da perda da superfície dentária devido à abrasão(AU)

Caracterização dos estágios iniciais do câncer colorretal utilizando microscopia de autofluorescência 3D: um estudo preliminar / Early stages of colorectal cancer characterization by autofluorescence 3d microscopy: a preliminary study

ABSTRACT Background: Colorectal cancer is one of the most prevalent pathologies worldwide whose prognosis is linked to early detection. Colonoscopy is the gold standard for screening, and diagnosis is usually made histologically from biopsies. Aiming to reduce the inspection and diagnostic time as well as the biopsies and resources involved, other techniques are being promoted to conduct accurate in vivo colonoscopy assessments. Optical biopsy aims to detect normal and neoplastic tissues analysing the autofluorescence spectrum based on the changes in the distribution and concentration of autofluorescent molecules caused by colorectal cancer. Therefore, the autofluorescence contribution analysed by image processing techniques could be an approach to a faster characterization of the target tissue. Objective: Quantify intensity parameters through digital processing of two data sets of three-dimensional widefield autofluorescence microscopy images, acquired by fresh colon tissue samples from a colorectal cancer murine model. Additionally, analyse the autofluorescence data to provide a characterization over a volume of approximately 50 µm of the colon mucosa for each image, at second (2nd), fourth (4th) and eighth (8th) weeks after colorectal cancer induction. Methods: Development of a colorectal cancer murine model using azoxymethane/dextran sodium sulphate induction, and data sets acquisition of Z-stack images by widefield autofluorescence microscopy, from control and colorectal cancer induced animals. Pre-processing steps of intensity value adjustments followed by quantification and characterization procedures using image processing workflow automation by Fiji's macros, and statistical data analysis. Results: The effectiveness of the colorectal cancer induction model was corroborated by a histological assessment to correlate and validate the link between histological and autofluorescence changes. The image digital processing methodology proposed was then performed on the three-dimensional images from control mice and from the 2nd, 4th, and 8th weeks after colorectal cancer chemical induction, for each data set. Statistical analyses found significant differences in the mean, standard deviation, and minimum parameters between control samples and those of the 2nd week after induction with respect to the 4th week of the first experimental study. This suggests that the characteristics of colorectal cancer can be detected after the 2nd week post-induction. Conclusion: The use of autofluorescence still exhibits levels of variability that prevent greater systematization of the data obtained during the progression of colorectal cancer. However, these preliminary outcomes could be considered an approach to the three-dimensional characterization of the autofluorescence of colorectal tissue, describing the autofluorescence features of samples coming from dysplasia to colorectal cancer.

RESUMO Contexto: O câncer colorretal é uma das patologias mais prevalentes em todo o mundo, cujo prognóstico está ligado à detecção precoce. A colonoscopia é o padrão ouro para triagem, e o diagnóstico geralmente é feito histologicamente a partir de biópsias. Visando reduzir o tempo de inspeção e diagnóstico, bem como as biópsias e recursos envolvidos, outras técnicas estão sendo promovidas para realizar avaliações precisas de colonoscopia in vivo. A biópsia óptica visa detectar tecidos normais e neoplásicos analisando o espectro de autofluorescência com base nas mudanças na distribuição e concentração de moléculas autofluorescentes causadas pelo câncer colorretal. Portanto, a contribuição da autofluorescência analisada por técnicas de processamento de imagem poderia ser uma abordagem para uma caracterização mais rápida do tecido-alvo. Objetivo: Quantificar parâmetros de intensidade por meio do processamento digital de dois conjuntos de dados de imagens de microscopia de autofluorescência em campo amplo tridimensionais, adquiridas por amostras de tecido fresco de cólon de um modelo murino de câncer colorretal. Adicionalmente, analisar os dados de autofluorescência para fornecer uma caracterização em um volume de aproximadamente 50 µm da mucosa do cólon para cada imagem, na segunda (2ª), quarta (4ª) e oitava (8ª) semanas após a indução do câncer colorretal. Método: Desenvolvimento de um modelo murino de câncer colorretal usando indução de azoximetano/sulfato de sódio dextrano, e aquisição de conjuntos de dados de imagens Z-stack por microscopia de autofluorescência em campo amplo, de animais controle e induzidos ao câncer colorretal. Etapas de pré-processamento de ajustes de valores de intensidade seguidas por procedimentos de quantificação e caracterização usando automação de fluxo de trabalho de processamento de imagem por macros do Fiji, e análise estatística de dados. Resultados: A eficácia do modelo de indução de câncer colorretal foi corroborada por uma avaliação histológica para correlacionar e validar a ligação entre as mudanças histológicas e de autofluorescência. A metodologia de processamento digital de imagem proposta foi então realizada nas imagens tridimensionais de camundongos controle e das 2ª, 4ª e 8ª semanas após a indução química do câncer colorretal, para cada conjunto de dados. Análises estatísticas encontraram diferenças significativas nos parâmetros médios, desvio padrão e mínimos entre amostras de controle e aquelas da 2ª semana após a indução em relação à 4ª semana do primeiro estudo experimental. Isso sugere que as características do câncer colorretal podem ser detectadas após a 2ª semana pós-indução. Conclusão: O uso de autofluorescência ainda apresenta níveis de variabilidade que impedem uma maior sistematização dos dados obtidos durante a progressão do câncer colorretal. No entanto, esses resultados preliminares podem ser considerados uma abordagem para a caracterização tridimensional da autofluorescência do tecido colorretal, descrevendo as características de autofluorescência de amostras que vão da displasia ao câncer colorretal.

Reuse of N95/PFF2 masks in clinical practice: morphological and structural analysis / Reutilización de mascarillas N95/PFF2 en la práctica clínica: análisis morfológico y estructural / Reutilização das máscaras N95/PFF2 na prática clínica: análise morfológica e estrutural

Objective: to analyze the integrity of N95/PFF2 masks in relation to fiber morphology, porosity, cracks and micro holes, as well as identify visible damage to their structure and components, after seven- and fifteen-day reuse protocols. Method: cross-sectional study. Structural and morphological characteristics of a new N95/PFF2 mask were analyzed in comparison with N95/PFF2 masks (n=10) used in seven- and fifteen-day protocols, through visual inspection and scanning electron microscopy. Results: upon visual inspection, following the seven-day protocol, 40% and 60% of the N95/PFF2 masks showed, respectively, personal identification marks and external and internal dirt. Additionally, 20% exhibited loosening and/or tearing of the straps, while 100% showed some type of damage to the nose clips. In the fifteen-day protocol, all N95/PFF2 masks had dirt, loose straps and damaged nose clips, and 80% had folds. Electronic microscopy revealed an increase in pores and loosening in the weaves from seven days onwards, extending up to fifteen days, with the presence of micro holes and residues. Conclusion: the reuse of N95/PFF2 masks affects their structural and morphological integrity. It is crucial to carry out tests to measure the impact of this practice on the safety of health professionals.

Objetivo: analizar la integridad de mascarillas N95/PFF2 con relación a la morfología de las fibras, porosidad, grietas y microagujeros, así como identificar daños visibles en su estructura y componentes, luego de protocolos de reutilización de siete y quince días. Método: estudio transversal. Se analizaron las características estructurales y morfológicas de una mascarilla N95/PFF2 nueva en comparación con las mascarillas N95/PFF2 (n=10) utilizadas en protocolos de siete y quince días, mediante inspección visual y microscopía electrónica de barrido. Resultados: tras la inspección visual, siguiendo el protocolo de siete días, el 40% y el 60% de las mascarillas N95/PFF2 mostraron, respectivamente, marcas de identificación personal y suciedad externa e interna. Además, el 20% presentó aflojamiento y/o desgarro de las correas de sujeción, mientras que el 100% presentó algún tipo de daño en los clips nasales. En el protocolo de quince días, todas las mascarillas N95/PFF2 tenían suciedad, correas de sujeción sueltas y clips nasales dañados, y el 80% tenía pliegues. La microscopía electrónica reveló un aumento de poros y aflojamiento en las tramas a partir de los siete días, extendiéndose hasta los quince días, con presencia de microagujeros y residuos. Conclusión: la reutilización de las mascarillas N95/PFF2 compromete su integridad estructural y morfológica. Es crucial realizar pruebas para medir el impacto de esta práctica en la seguridad de los profesionales de la salud.

Objetivo: analisar a integridade das máscaras N95/PFF2 em relação à morfologia das fibras, porosidade, fissuras e micro furos, bem como identificar danos visíveis em sua estrutura e componentes, após protocolos de reutilização de sete e quinze dias. Método: estudo transversal. Características estruturais e morfológicas de uma máscara N95/PFF2 nova foram analisadas em comparação com máscaras N95/PFF2 (n=10) utilizadas em protocolos de sete e quinze dias, por meio da inspeção visual e microscopia eletrônica de varredura. Resultados: na inspeção visual, seguindo o protocolo de sete dias, 40% e 60% das máscaras N95/PFF2 apresentaram, respectivamente, marcas de identificação pessoal e sujidades externas e internas. Além disso, 20% exibiram afrouxamento e/ou rompimento das tiras de fixação, enquanto 100% mostraram algum tipo de dano nos clipes nasais. No protocolo de quinze dias, todas as máscaras N95/PFF2 apresentavam sujidade, tiras de fixação frouxas e clipes nasais comprometidos; 80% possuíam dobras. A microscopia eletrônica revelou aumento dos poros e afrouxamento nas tramas a partir de sete dias, ampliando-se até quinze dias, com presença de micro furos e detritos. Conclusão: a reutilização das máscaras N95/PFF2 compromete a integridade estrutural e morfológica. É crucial realizar testes para mensurar o impacto dessa prática na segurança dos profissionais de saúde.

Synthesis, characterization and protection effect of black rice anthocyanins nano-composite against hepatotoxicity induced by methotrexate in rats

Abstract The present study aimed to investigate the beneficial of prepared black rice anthocyanins nano-composite (An-AgNps) against hepatotoxicity induced by methotrexate (MTX) in rats. Anthocyanins nano-composite was prepared by silver as the metallic ion reduction and were characterized by IR and SEM. The rats in our experiment were divided into five groups. Serum lipid profile, serum transaminases (ALT and AST), ALP, LDH, TBA, GSH and SOD were examined. The results show that SEM of An-AgNps has average particle size from 70 to 130nm. In the group treated with MTX; TC, TG, LDL-c, ALT, AST, ALP, LDH and TBA levels were significantly (P0.05) increased than NC, while, HDL-c, SOD and GSH levels were significantly (P0.05) decreased. On the other hand, An-AgNps + MTX treated groups were reversed the levels of all biomarkers similar to NC. In conclusion, the results show that An-AgNps has a protective effect on MTX-induced hepatotoxicity and oxidative stress.

Resumo O presente estudo teve como objetivo investigar o benefício de nanocompósito de antocianinas de arroz preto preparado (An-AgNps) contra a hepatotoxicidade induzida por metotrexato (MTX) em ratos. O nanocompósito de antocianinas foi preparado a partir da prata por meio da redução do íon metálico e caracterizado por IR e SEM. Os ratos em nosso experimento foram divididos em cinco grupos, e foram examinados o perfil lipídico sérico, as transaminases séricas (ALT e AST), ALP, LDH, TBA, GSH e SOD. Os resultados mostram que SEM de An-AgNps tem tamanho médio de partícula de 70 a 130 nm. No grupo tratado com MTX, os níveis de TC, TG, LDL-c, ALT, AST, ALP, LDH e TBA aumentaram significativamente (P 0,05) do que NC, enquanto os níveis de HDL-c, SOD e GSH diminuíram significativamente (P 0,05). Por outro lado, nos grupos tratados com An-AgNps + MTX, foram revertidos os níveis de todos os biomarcadores semelhantes ao NC. Em conclusão, os resultados mostram que o An-AgNps tem um efeito protetor contra a hepatotoxicidade induzida pelo MTX e o estresse oxidativo.

Effectiveness of endoscopy in cochlear implantation.

OBJECTIVE: Determine the effectiveness of endoscopy in cochlear implantation as compared to microscopy. METHOD: Study comparing microscopy and endoscopy in cochlear implant placement in 34 patients (23 endoscopic implants and 20 implants via microscopy), between 2014 and 2019, at the Centro Medico Naval, Mexico City. The study was performed under informed consent and according to the Council for International Organizations of Medical Sciences (CIOMS). RESULTS: Of the 34 patients, 12 were children or adolescents and 22 were adults. The visualization of the round window classified via microscopy per St. Thomas Hospital's classification showed that type IIB prevailed in 30.2% of patients, and type III in 41.9%, and when using the endoscope, the round window was observed in full in 82.6% of patients (type I), and type IIA was only observed in 17.4% (four patients). The number of attempts made to place the cochlear implant was greater with the microscope. The time to insertion of the electrode was 1.6 minutes. No differences were observed (p > 0.05) in the number of inpatient days. Cochleostomy was more frequent when using the microscope. CONCLUSIONS: Endoscopy is an effective resource in cochlear implantation for posterior tympanotomy, with no complications observed, offering greater safety in inserting the electrode through the round window.

OBJETIVO: Determinar la efectividad de la endoscopía en la implantación coclear en comparación con la técnica microscópica. MÉTODO: Se comparó la microscopía frente a la endoscopía en la colocación de implante coclear en 34 pacientes (23 endoscópicos y 20 microscópicos), del año 2014 al año 2019, en el Centro Médico Naval de la Ciudad de México. El estudio se realizó bajo consentimiento informado y apegado a las normas del Council for International Organizations of Medical Sciences. RESULTADOS: De los 34 pacientes, 12 eran niños o adolescentes y 22 eran adultos. La visualización de la ventana redonda fue clasificada con microscopio según la clasificación del St. Thomas Hospital, predominando la tipo IIB (30.2%) y la III (41.9%), y al utilizar el endoscopio se observó completa en el 82.6% (tipo I) y tipo IIA en tan solo el 17.4% (cuatro pacientes). El número de intentos en la colocación del implante coclear fue mayor con el microscopio. El tiempo en el que se insertó el electrodo fue de 1.6 minutos. No hubo diferencias (p > 0.05) en la estancia hospitalaria. Fue más frecuente la cocleostomía cuando se uso el microscopio. CONCLUSIONES: La endoscopía es un instrumento efectivo en la implantación coclear por timpanotomía posterior, sin presentarse complicaciones y dando mayor seguridad para insertar el electrodo por la ventana redonda.

Displasia mesenquimal placentaria

La displasia mesenquimal placentaria es una entidad poco frecuente, confundida al ultrasonido y macroscopía con mola parcial por las lesiones vesiculares y la presencia de feto. La microscopía revela vellosidades troncales hidrópicas con gran hiperplasia mesenquimal y lesiones vasculares prominentes, pero sin hiperplasia del epitelio trofoblástico. El feto, generalmente femenino, puede ser normal o presentar retardo del crecimiento intrauterino, malformaciones, tumores o cromosomopatías. Presentamos el estudio de cuatro casos de displasia mesenquimal placentaria; tres primigestas de 39 y 20 años de edad, y una segundigesta de 26 años. La mayor, del tercer trimestre, cursó con preeclampsia y mortinato de sexo femenino con anomalías pulmonar y esplénica, las otras dos presentaron sangrado vaginal y dolor pélvico y un caso fue un hallazgo en el control prenatal a las 8 semanas de gestación. En tres casos se observaron feto y embrion, dos de ellos tuvieron necropsia.

Placental mesenchymal dysplasia is a rare entity, confused with ultrasound and macroscopy with partial mole due to vesicular lesions and the presence of a fetus. Microscopy reveals hydropic trunk villi with great mesenchymal hyperplasia and prominent vascular lesions but without hyperplasia of the trophoblastic epithelium. The fetus, usually female, may be normal or present malformations, tumors or chromosomopathies. We present the study of four cases of placental mesenchymal dysplasia; three first-pregnancies, aged 39 and 20, and a second-pregnancy, aged 26. The oldest in the third trimester presented with preeclampsia and a female stillbirth with pulmonary and splenic anomalies, the other two presented vaginal bleeding and pelvic pain and one case was a finding in prenatal control at 8 weeks of gestation. In three cases, fetus and embryo were observed, two of them had a necropsy.