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.

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.

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.

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.

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.

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.

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.

Avaliação dos Efeitos da Remoção da Resina Residual e do Polimento na Superfície do Esmalte após Descolagem do Braquete / Evaluation of the Effects of Residual Resin Removal and Polishing on the Enamel Surface after Bracket Debonding

Considerando o uso de brocas para remoção da resina residual após descolagem do braquete e a possibilidade de injúrias à superfície do esmalte após o uso dessas brocas, este trabalho teve como objetivo realizar um estudo experimental, para avaliar a variação do aspecto superficial do esmalte de forma qualitativa, por meio da avaliação com imagens topográficas do esmalte dentário, utilizando-se a Microscopia Eletrônica de Varredura (MEV), a qual permitiu ilustrar e avaliar a superfície do esmalte após a fase de polimento final, realizada por dois métodos: taça de borracha ou escova Robinson. Foram utilizados 25 dentes pré-molares humanos, obtidos a partir de exodontias em pacientes que procuraram voluntariamente o curso de Residência em Cirurgia da Clínica Odontológica Universitária da Universidade Estadual de Londrina. Os dentes foram divididos em quatro grupos: A, B, C e D, contendo 6 dentes cada, de acordo com as brocas utilizadas para a remoção do remanescente adesivo e o polimento escolhido, além de um dente como "controle". Foi avaliada a rugosidade superficial do esmalte após a remoção da resina e a superfície do esmalte após o polimento com as duas opções apresentadas. Os resultados mostraram que, por observação e inspeção, as brocas removeram a resina residual de todos os dentes, porém, causaram riscos e ranhuras, como evidenciado nas imagens em MEV. Concluiu-se que não houve diferença estatística entre os métodos de polimento e que ambos foram importantes para a redução das marcas abrasivas, proporcionando uma superfície mais lisa do esmalte.

Considering the use of specific burs to remove residual resin after bracket debonding and the possibility of injuries to the dental enamel after using these burs, this study aimed to verify the variation in the enamel surface appearance in a qualitative way and evaluation with topographic images of the dental enamel. The use of Scanning Electron Microscopy (SEM) allowed to illustrate and evaluate the enamel surface after the final polishing phase using two methods: rubber cup or Robinson brush. Twenty-five human premolar teeth were obtained from extractions in patients who voluntarily sought the Oral Maxillofacial Surgery Residency at the Dental School from the State University of Londrina; the teeth were divided into four groups A, B, C and D containing 6 teeth each according to the burs used to remove the remaining adhesive and the chosen polishing, in addition to one tooth as a "control". Dental enamel surface roughness was evaluated after resin removal and enamel surface after polishing with the two methods presented. The results showed that by observation and inspection, the burs removed residual resin from all teeth, however, caused scratches and grooves as evidenced in the SEM images. Based on the results, there was no statistical difference between the polishing methods, and both were important for the reduction of abrasive marks and provided a smoother enamel surface.

Effect of different obturation techniques on sealer penetration into dentinal tubules after final irrigation with XP-Endo finisher file

The first objective of this research was to evaluate the effectiveness of XP-Endo Finisher on dentinal tubule penetration of irrigation solution using confocal laser scanning microscopy. The main purpose of this research was to compare the effect of cold lateral condensation, continuous wave obturation and core-carrier based techniques on sealer penetration. Sixty mandibular premolars were prepared and allocated into two experimental groups (n=30) as the final irrigation technique and obturation technique experiment. In the final irrigation technique experiment, final irrigation was performed with XP-Endo Finisher, passive ultrasonic irrigation (PUI) and conventional needle irrigation (CNI) (n=10). The roots in the obturation technique experiment were also assigned into 3 groups and obturated with cold lateral condensation, continuous-wave obturation and core-carrier techniques (n=10). The most effective activation method, which emerged as a result of the first part of this study, was used as the final irrigation method in the obturation technique experiment. Then, all roots were sectioned in 1-mm-thick slices at 3mm from the apex for scanning. In terms of depth and percentage of material penetration, CNI exhibited significantly the lowest values and no significant difference was found between others. Also, there was no significant difference among obturation methods. In conclusion, XP-Endo Finisher and PUI are more effective than CNI on irrigant penetration. Sealer penetration into dentinal tubules is independent of obturation techniques.

El objetivo principal de esta investigación fue evaluar la eficacia de XP- Endo Finisher en la penetración de la solución de irrigación en los túbulos dentinarios mediante microscopía de láser confocal. Se prepararon sesenta premolares mandibulares y se distribuyeron en dos grupos experimentales (n=30) según el tipo de método de evaluación utilizado. En el experimento de la técnica de irrigación final, la irrigación final se realizó con XP-Endo Finisher, irrigación ultrasónica pasiva (PUI) e irrigación con aguja convencional (CNI) (n=10). Las raíces en el experimento de la técnica de obturación también se asignaron en 3 grupos y se obturaron con técnicas de condensación lateral fría, obturación de onda continua y portador de núcleo (n=10). El método de activación más eficaz, que surgió como resultado de la primera parte de este estudio, se utilizó como método de irrigación final en el experimento de la técnica de obturación. Luego, todas las raíces se seccionaron en muestras de 1mm de espesor. En términos de profundidad y porcentaje de penetración del material, CNI exhibió significativamente los valores más bajos y no se encontraron diferencias significativas entre los demás. Además, no hubo diferencias significativas entre los métodos de obturación. En conclusión, XP-Endo Finisher y PUI son más efectivos que CNI en la penetración del irrigante. La penetración del sellador en los túbulos dentinarios es independiente de las técnicas de obturación.

Descompresión microvascular endoscópica versus microscópica en neuralgia trigeminal en un centro Argentino / Endoscopic vs. microscopic techniques for the treatment of trigeminal neuralgia. Our experience at an Argentine care center

Resumen Introducción: La descompresión microvascular (DMV) en la neuralgia trigeminal es una técnica quirúrgica cuyo objetivo es revertir la compresión a la que se ve someti do un nervio por una estructura vascular. El objetivo de este estudio fue realizar una comparación directa entre la descompresión microvascular endoscópica (DMV-E) y la misma a través del uso del microscopio (DMV-M) en el tratamiento de la neuralgia del trigémino. Métodos: Se realizó un estudio de cohorte retrospec tivo de pacientes operados de neuralgia de trigémino, por un mismo cirujano, entre 2015 y 2021 en nuestra institución, tanto por técnica microquirúrgica como endoscópica. Resultados: Se obtuvieron un total de 31 pacientes divididos en dos grupos: Grupo M correspondiente a 15 (49%) pacientes abordados con técnica microscópica y Grupo E, con 16 (51%) pacientes intervenidos con técnica endoscópica. Se identificaron diferencias en el tamaño de la cra niectomía, más pequeña en el grupo E (2.50 cm vs 3.70 cm grupo M); y en el tiempo de internación, de 2.43 días en el grupo E vs. 4.46 días en el grupo M. El tiempo de cirugía fue similar para ambas técnicas quirúrgicas La principal compresión fue dada por la arteria ce rebelosa superior (ACS) en ambos grupos. Todos los pacientes presentaron mejoría del Barrow Neurological Institute Pain Intensity Score (BNI) en el postoperatorio en ambos grupos. Discusión: La DMV-E constituye una alternativa qui rúrgica interesante a la ya conocida DMV-M para el tratamiento de la neuralgia trigeminal, por requerir menores dimensiones en la incisión cutánea y tamaño de la craniectomía, acortando el tiempo de internación, lo cual no solo implica un beneficio para el paciente, sino que también representa menor costo de internación.

Abstract Introduction: Trigeminal neuralgia is a highly invali dating pathology, whose natural course has been modi fied thanks to decompressive microvascular surgery. The intervention can be carried out either with a microscope or via an endoscopic technique. Our goal was to compare these two techniques for the treatment of this complex pathology. Methods: Retrospective, analytical study of a cohort of patients treated by a single surgeon at our institution, in the period between 2015 and 2021. Results: We identified 31 patients and divided them into two groups: 15 (49%) treated using the microscopic technique (group M), and 16 (51%) exclusively via an endoscopic one (group E). Differences were observed between the means of the size of the craniectomy in group M (3.7 cm) compared to group E (2.5 cm); The mean length of hospital stay for patients in group E was shorter (4.46 days compared to that of patients in group M, whose hospital stay averaged 2.43 days). There were no differences between the two groups regarding the length of the procedure. In both groups, the predomi nant compression was due to the superior cerebellar artery (SCA). Pain outcomes were equivalent, with every patient in both groups having an improved postoperative Barrow Neurological Institute Pain Intensity Score (BNI). Discussion: Endoscopic microvascular decompression is an attractive option for the resolution of neurovas cular conflict as it provides functional results similar to the microscope technique, without requiring an exten sive craniectomy and associated to shorter in-hospital stay, which is beneficial for both the patient and the institution.

Light Microscopic, Ultrastructure Analysis and Functional Morphology of Cornu aspersum Spermatozoa Contained in the Frozen Hermaphroditic Duct / Análisis de Microscopía de Luz, Ultraestructura y Morfología Funcional de Espermatozoides de Cornu aspersum Contenidos en el Conducto Hermafrodita Congelado

SUMMARY: In this study we describe the functional morphology of Cornu aspersum (Helix aspersa), spermatozoa using light, scanning (SEM) and transmission electron (TEM) microscopies. The studies were performed with sperm located in the frozen hermaphroditic duct. Our results showed that the head presents an elongated conical shape slightly coiled in a corkscrew, with the nucleus partially covered by an acrosome, where an apical vesicle is located at the lateralized apex. This peculiar shape suggests the helical displacement movement of the spermatozoa. The head and the nucleus are slightly larger size compared to those of other gastropod species. The intermediate tract is surrounded by a mitochondrial complex and a glycogen helix. The glycogen helix is coiled helically along the intermediate tract, presenting at least five twists of glycogen helices. The complexity of both the mitochondrial complex and the glycogen helix suggests a high metabolic consumption considering the long period of time until fertilization occurs. Our findings on the detailed characterization of Cornu aspersum spermatozoa, obtained from a frozen hermaphroditic duct can contribute to a better understanding of the functional morphology of sperm and serve as a reference for future studies.

En este estudio describimos la morfología funcional de Cornu aspersum (Helix aspersa), espermatozoides utilizando microscopías de luz, barrido (SEM) y electrónica de transmisión (TEM). Los estudios se realizaron con espermatozoides localizados en el conducto hermafrodita congelado. Nuestros resultados mostraron que la cabeza presenta una forma cónica alargada ligeramente enrollada en un tirabuzón, con el núcleo parcialmente cubierto por un acrosoma, donde se ubica una vesícula apical en el ápice lateralizado. Esta peculiar forma sugiere el movimiento de desplazamiento helicoidal de los espermatozoides. La cabeza y el núcleo son de un tamaño ligeramente mayor en comparación con los de otras especies de gasterópodos. El tracto intermedio está rodeado por un complejo mitocondrial y una hélice de glucógeno. La hélice de glucógeno se enrolla helicoidalmente a lo largo del tracto intermedio, presentando al menos cinco giros de hélices de glucógeno. La complejidad tanto del complejo mitocondrial como de la hélice de glucógeno sugiere un alto consumo metabólico considerando el largo período de tiempo hasta que ocurre la fecundación. Nuestros hallazgos sobre la caracterización detallada de los espermatozoides de Cornu aspersum, obtenidos de un conducto hermafrodita congelado, pueden contribuir a una mejor comprensión de la morfología funcional de los espermatozoides y servir como referencia para futuros estudios.

Ayudas diagnósticas y diagnóstico diferencial de la enfermedad de Pompe / Diagnostic aids and differential diagnosis of Pompe disease

Introducción: La enfermedad de Pompe (EP) o glucogenosis tipo II es una enfermedad autosómica recesiva causada por mutaciones en el gen GAA que codifica para la proteína alfa-1,4-glucosidasa. Su deficiencia lleva a un almacenamiento anormal de glucógeno en los lisosomas de varias células, a través de los diferentes tejidos, lo que causa un compromiso musculoesquelético predominante. Contenidos: Los fenotipos de la enfermedad dependen de las variantes genéticas y de los niveles de la actividad enzimática residual. La enfermedad se presenta como EP de inicio infantil, EP de inicio tardío y EP intermedio, por lo que es de suma importancia su diagnóstico temprano, por medio de estudios moleculares como la secuenciación de Sanger y la secuenciación de nueva generación. Conclusiones: Se ha demostrado, mediante diferentes estudios, que las variaciones genéticas pueden diferir entre etnias, y es importante su caracterización molecular para determinar el tratamiento más adecuado, de acuerdo con el estado del material inmunológico de reacción cruzada (CRIM).

Introduction: Pompe disease (PD) or Glycogenosis Type II is a rare autosomal recessive disease caused by mutations in the GAA gene that codes for the alpha-1,4-glucosidase protein. Its deficiency leads to abnormal glycogen storage in the lysosomes of various cells throughout the different tissues causing a predominant musculoskeletal compromise. Contents: The phenotypes of the disease depend on the genetic variants and the levels of residual enzyme activity, presenting as infantile-onset PD, late-onset PD, and intermediate PD; Therefore, early diagnosis of the disease through molecular studies such as Sanger sequencing and new generation sequencing is of utmost importance. Conclusions: It has been shown through different studies that genetic variations can vary between ethnic groups and the molecular characterization of the variants is important to determine the most appropriate treatment depending on the state of the cross-reactive immunological material (CRIM)