Histomorphological, Histochemical and Ultrastructural Studies on the Healthy Liver of Yemen Veiled Chameleon (Chamaeleo calyptratus) in Southern Saudi Arabia / Estudios Histomorfológicos, Histoquímicos y Ultraestructurales en el Hígado Sano del Camaleón Velado de Yemen (Chamaeleo calyptratus) en el Sur de Arabia Saudita

SUMMARY: The livers of reptiles are being studied as a model for the link between the environment and hepatic tissue. There have been few investigations on the histology of reptile livers, and very few or no studies have examined the histology of liver of veiled chameleon (Chamaeleo calyptratus). This paper describes the histomorphological, histochemical and ultrastructural characterization of the liver of veiled chameleons in southern Saudi Arabia. Seven Chamaeleo calyptratus were captured in the summer season in Abha City, Aseer region, southern Saudi Arabia. Chamaeleon liver samples were processed for histomorphology, histochemistry and ultrastructure analyses. Morphologically liver of Chamaeleo calyptratus was observed as a large dark brown organ with lighter speckles, which represent melanin deposits. It located at the ventral part of abdominal cavity forward of the stomach. Its dimensions approximately were 3.7 x 2 cm. The liver was a bilobed organ divided into two lobes, right and left lobes. The right one was bigger than the others. The gallbladder was well developed and had an elongated shape, situated between the two lobes and contained the bile for the digestion. Microscopically, the liver was found to be covered by a thick layer of connective tissue, which formed the hepatic capsule. Hepatic parenchyma probably appeared in cross sections as hepatic glandular-like alveoli "acini" or follicular structures with various diameters, each acinus contains approximately four to six hepatocytes, surrounded by sinusoidal capillaries filled with abundant melanomacrophages, which are absent in birds and mammals. Melanomacrophages are common in the hepatic parenchyma's perisinusoidal areas, particularly near portal spaces. Hepatocytes are polyhedral or pyramidal with and mostly contained large, rounded nuclei mostly peripherally located, with prominent dark oval nucleoli. Some of nuclei are eccentric or central position. The cytoplasm appeared spongy or vacuolated and more eosinophilic when stained by hematoxylin-eosin and strongly reactive to PAS staining technique, indicating abundant glycogen content. The reticular fibers that surround hepatocytes, blood arteries, and sinusoids supported the hepatic parenchyma. The blood sinusoids are seen interspersed among hepatocytes of varying sizes. The sinusoidal lumen was bordered by flattened endothelial cells and includes elliptical nucleated erythrocytes and liver macrophages as phagocytes, which are also known as Kupffer cells. Branches of the portal vein, hepatic artery, small bile duct, and lymph vessels were detected in the hepatic portal area "tract" or triad which made up of connective. Hematopoietic tissue was observed in subcapsular region and portal triads. Ultrastructurally, the hepatocyte appeared polyhedric containing a single large rounded basal or eccentric vesicular nucleus with prominent nucleolus. Extensive network of rough endoplasmic reticulum (RER) often arranged in an array parallel to the nuclear membrane with many mitochondria, and Golgi apparatus were described. The cytoplasm contained glycogen granules, vesicles or vacuoles scattered throughout the cytoplasm especially at the apical region were reported. The bile canaliculi and the hepatic "Kupffer" cells were also discussed. This is the first study on the histological characterization of the healthy liver of Yemen veiled chameleon in southern Saudi Arabia. The findings reported here should be used as a reference to compare with the pathological abnormalities of the liver in this animal.

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Avaliação do comportamento das fibras colágenas periodontais durante a progressão da periodontite experimental em ratos / Evaluation of the behaviour of periodontal collagen fibers during the progression of experimental periodontitis in rats

Introdução: A periodontite é uma doença infecto-inflamatória, resultante da disbiose microbiana e da resposta do hospedeiro, que leva à destruição dos tecidos de suporte dentário, inclusive das fibras colágenas periodontais, podendo culminar na perda do elemento dental. Objetivo: Avaliar o comportamento das fibras colágenas periodontais durante a progressão da periodontite experimental induzida em ratos. Material e método: Doze ratos Wistar foram distribuídos nos grupos: Controle (C), Periodontite Experimental 14-dias (PE-14d), Periodontite Experimental 21-dias (PE-21d) e Periodontite Experimental 42-dias (PE-42d). No dia 0, os animais do grupo C foram eutanasiados. Neste mesmo dia, os animais remanescentes foram submetidos à instalação de uma ligadura de algodão ao redor do primeiro molar inferior esquerdo para indução da periodontite experimental. Tais animais foram eutanasiados aos 14 (PE-14d), 21 (PE-21d) e 42 (PE-42d) dias após a instalação da ligadura. Executou-se o processamento histológico das hemimandíbulas e as secções foram submetidas à reação histoquímica pelo vermelho picro-sirius. A análise qualitativa descritiva foi realizada sob microscopia de luz polarizada, na região de furca dental, evidenciando as fibras do ligamento periodontal. Resultado: O grupo C exibiu feixes espessos e orientados de fibras colágenas maduras, condizentes com aspecto de normalidade. Os grupos com periodontite experimental exibiram desestruturação tecidual severa, com fibras colágenas imaturas e de menor espessura, sendo tais condições mais exacerbadas nos grupos PE-14d e PE-21d. Conclusão: As fases iniciais da periodontite apresentam caráter agudo e, portanto, resultam na rápida destruição dos tecidos periodontais de suporte, prejudicando potencialmente a fibrilogênese e a reestruturação do colágeno no ligamento periodontal.

Introduction: Periodontitis is an infectious-inflammatory disease resulting from microbial dysbiosis and host response that leads to the destruction of tooth support tissues, including periodontal collagen fibers, which may culminate in tooth loss. Objective: To evaluate the behavior of periodontal collagen fibers during the progression of induced experimental periodontitis in rats. Material and method: Twelve Wistar rats were distributed into groups: Control (C), 14-days Experimental Periodontitis (PE-14d), 21-days Experimental Periodontitis (PE-21d) and 42-days Experimental Periodontitis (PE-42d). At day 0, the animals of group C were euthanized. At the same day, the remaining animals were submitted to the installation of a cotton ligature around the lower left first molar for the induction of experimental periodontitis. The animals were euthanized at 14 (PE-14d), 21 (PE-21d) and 42 (PE-42d) days after the installation of ligature. Histological processing of the hemi-mandibles was performed and the sections underwent histochemical reaction using picro-sirius red. The descriptive qualitative analysis was performed under polarized light microscopy, in the dental furcation region, evidencing the fibers of the periodontal ligament. Result: Group C exhibited thick and oriented bundles of mature collagen fibers, consistent with a normal appearance. The groups with experimental periodontitis exhibited severe tissue disruption, with immature and thinner collagen fibers, with such conditions being more exacerbated in the PE-14d and PE-21d groups. Conclusion: The early stages of periodontitis present acute response, and therefore result in rapid destruction of periodontal support tissues and potentially impair fibrillogenesis and collagen restructuring in the periodontal ligament.

Incidencia de fracturas dentales reportadas en la Ciudad de Chihuahua durante la pandemia por COVID-19 / Incidence of dental fractures reported in Chihuahua city during COVID-19 pandemic

Se presentan los casos de fracturas verticales sintomáticos que ameritaron la extracción de las órganos dentales afectados en un periodo de cinco meses durante la pandemia por COVID-19 en la ciudad de Chihuahua, México. Material y métodos: Se recolectaron 26 muestras para el estudio, proporcionadas por los cirujanos dentistas integrantes del Grupo de Egresados de la Facultad de Odontología de la Universidad Autónoma de Chihuahua. Resultados: Los resultados muestran diferencias a lo reportado previamente respecto al tipo de diente involucrado, sexo de los pacientes, restauraciones coronales y tratamientos endodóncicos previos. Conclusiones: Se puede suponer que las modificaciones en el estilo de vida de la población, sumadas a la angustia permanente, pueden ser factores que contribuyen a las fracturas dentales (AU)

There are cases of symptomatic vertical fractures that warranted the extraction of affected teeth are presented, over a period of five months during the COVID-19 pandemic in Chihuahua City, Mexico. Material and methods: 26 samples were collected for the study, provided by the dentist who were members of the Graduate Group of the Faculty of Dentistry of the Autonomous University of Chihuahua. Results: The results show differences from what was previously reported regarding the type of tooth involved, sex of patients, coronal restorations and previous endodontic treatments. Conclusions: It can be assumed that changes in the lifestyle of the population, added to permanent distress, may be factors to preserve dental fractures (AU)

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

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

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

Detección y cuantificación de células plasmáticas (plasmocitos) en granulomas periapicales inducidos experimentalmente en ratas / Detection and quantification of plasma cells in experimentally induced rat periapical granulomas

Objetivo: En los granulomas periapicales, los plasmocitos (PL) participan activamente mediante la liberación de inmunoglobulinas. El propósito de este ensayo fue identificar y contar el número de PL en diferentes períodos de tiempo en lesiones periapicales experimentales en ratas. Materiales y métodos: Mediante la exposición al medio oral de la pulpa de los primeros molares inferiores izquierdos, se indujeron granulomas periapicales en ratas a las que previamente se les suministró anestesia. La pulpa de los primeros molares inferiores derechos no fue expuesta, y estos dientes se utilizaron como control. Los animales fueron eutanasiados a los 10, 30 y 60 días de la exposición. Los maxilares inferiores fueron removidos, y los primeros molares, junto con los tejidos circundantes, se procesaron para su estudio histológico. Se obtuvieron secciones semiseriadas, posteriormente coloreadas con verde de metilo-pironina (VMP). Cada tres secciones, las tres siguientes fueron coloreadas con hematoxilina y eosina (H-E). Los controles también fueron coloreados con H-E. Resultados: Todos los especímenes experimentales coloreados con H-E revelaron la presencia de granulomas periapicales. Luego de la exposición pulpar, el número de PL que reaccionó positivamente al VMP se incrementó de manera progresiva desde el día 10 hasta los días 30 y 60. A pesar de que a los 60 días el número de PL fue ligeramente menor que a los 30 días, no hubo diferencias estadísticamente significativas entre estos dos períodos. Los especímenes del grupo control coloreados con H-E mostraron que los tejidos periapicales se encontraban dentro de los parámetros normales en todos los períodos de observación. Conclusiones: Los resultados de este estudio revelaron que el número de plasmocitos VMP positivos se incrementa progresivamente en función del tiempo transcurrido pero se estabiliza al finalizar el experimento. También sugieren que el empleo de la coloración de VMP es un procedimiento adecuado para la identificación y la cuantificación de plasmocitos en los granulomas periapicales inducidos experimentalmente en ratas (AU)

Aim: Plasma cells (PL) release immunoglobulin in periapical lesions. The purpose of this assay was to identify and count the number of plasmocytes observed in periapical lesions in rats. Materials and methods: By exposing the pulp of the lower left first molars to the oral environment, periapical granulomas were induced in rats previously anesthetized. The pulp of right mandibular first molars was not exposed and these teeth were used as negative controls. The animals were euthanized at 10, 30 and 60 days after pup exposure. The mandibles were removed and specimens of the molar teeth along with the surrounding tissues were prepared for histology. Semi serial sections of the left first molar were stained with methyl green pyronine (MGP). Every three sections, the following three sections were stained with hematoxilyn and eosin (H-E). Negative control samples were stained with H-E. Results: All the H-E stained experimental samples revealed the presence of periapical granulomas. After pulp exposure, the number of PL increased from day 10 to 30 and 60. In the 60-day samples the number of PL was slightly less than that of the 30-day samples, with no statistically significant difference. The H-E stained control samples showed normal periapical tissues in all observation periods. Conclusions: The results of this study revealed that the number of VMP positive PL, increased progressively with time but it was stabilized at the end of the experiment. In addition, the results suggest that the use of VMP stain is a suitable procedure for the identification and counting of PL in experimentally induced periapical granulomas in rats (AU)

Micrografía analítica de semillas de Cebil (Anadenanthera colubrina var. Cebil (Griseb.) Altschul, Fabaceae) y Chamico (Datura ferox L., Solanaceae), empleadas como drogas alucinógenas en Argentina / Analytical micrography of seeds of Cebil (Anadenanthera colubrina var. Cebil (Griseb.) Altschul, Fabaceae) and Chamico (Datura ferox L., Solanaceae), employed as hallucinogenic drugs in Argentina

Resumen La micrografía analítica es una herramienta útil para la identificación de restos vegetales en muestras de material trozado o molido que no podrían ser identificadas de forma morfológica. El objetivo de este estudio es conocer los caracteres mi- crográficos de las semillas de Cebil (Anadenanthera colubrina var. cebil (Griseb.) Altschul, Leguminosae) y Chamico (Datura ferox L., Solanaceae), con el fin de proporcionar parámetros empleables para su identificación en un contexto forense y toxicológico. Los caracteres micrográficos relacionados con el tegumento exterior y las esclereidas fueron los más indicados para diferenciar entre ambas especies.

Abstract Analytical micrography is a useful tool for the identification of plant parts that can't be identified for its morphological characters. The aim of this work is to obtain micrographic characters of Cebil (Anadenanthera colubrina var. cebil (Griseb.) Altschul, Leguminosae) and Chamico (Datura ferox L., Solanaceae) seeds for its identification in a toxicological and forensic context. The most suitable micrographic features were the ones related with exterior testa and the stone cells.

Estudo da correlação das proteínas INGs com a P53 em lesões odontogênicas epiteliais benignas / Study of the correlation of INGs proteins with P53 in benign epithelial odontogenic lesions

As proteínas INGs (inhibitor of growth gene) desempenham papel de supressoras tumorais e podem agir por vias dependentes, ou independentes, da p53 na sinalização do ciclo celular e da apoptose. Este trabalho investigou, por meio de imuno-histoquímica, a correlação entre a expressão das proteínas INGs e a expressão da proteína p53 em ceratocistos odontogênicos (20), TOAs (20) e ameloblastomas sólidos (20). Os espécimes foram submetidos à marcação utilizando os anticorpos anti-Ing3, anti-Ing4, anti-Ing5 e anti-p53. Foi realizada análise quantitativa levando-se em consideração a localização citoplasmática e/ou nuclear para as proteínas INGs e a localização nuclear para a proteína p53. A análise da imunoexpressão das proteínas ING1 e ING2 foi realizada em um estudo prévio e os resultados foram considerados apenas para a análise de correlação com as proteínas estudadas neste estudo. Os dados foram analisados pelo Statistical Package for Social Sciences para Windows (SPSS versão 22.0; IBM, USA). Para a comparação da imunoexpressão entre os grupos de lesões foi utilizado o teste de Kruskal Wallis, e para a investigação das correlações foi utilizado o teste de Spearman. Foram considerados significativos os valores de p ≤ 0.05. O presente estudo evidenciou redução da expressão nuclear e citoplasmática das proteínas ING3, ING4 e ING5 em ceratocistos odontogênicos (COs) e ameloblastomas (AMBs). Além disso, em alguns casos, a perda da expressão nuclear das INGs esteve negativamente correlacionada à expressão da proteína p53. As análises de correlação entre as proteínas INGs indicam a existência de mecanismos compensatórios entre as proteínas INGs em folículos dentários (FDs) e tumores odontogênicos adenomatoides (TOAs), estes mecanismos parecem ser menos evidentes em COs e AMBs. Observou-se redução na expressão da proteína ING3 em AMBs (p=0,003); redução na expressão da proteína ING4, tanto em AMBs (p=0,02) quanto em COs (p=0,001); e uma redução da expressão nuclear da proteína ING5 nos COs (p=0,09) e nos AMBs (p=0,012). Foram evidenciadas correlações positivas entre a expressão nuclear da p53 com a expressão citoplasma/núcleo da proteína ING1 (r=0,603; p=0,05) em COs, e com a expressão citoplasma/ núcleo das proteínas ING3 (r=0,475; p=0,034) e ING4 (r=0,448; p=0,047) em AMBs. Por fim, os resultados deste estudo sugerem que a redução na expressão nuclear das proteínas INGs pode ser um evento envolvido na etiopatogênese de lesões odontogênicas mais agressivas, e que a redução da expressão nuclear/citoplasmática das proteínas INGs não está relacionada ao aumento expressão da p53 em COs e AMBs, o que sugere que a expressão destas proteínas deve resultar em alterações funcionais de maneira independente da p53 em lesões odontogênicas (AU).

INGs (inhibitor of growth gene) proteins play a role of tumor suppressors and can act via p53-dependent or independent pathways in signaling cell cycle and apoptosis. The aim of this study is to evaluate correlation between expression of proteins of ING proteins and expression of protein p53 in dental follicles (DF), odontogenic keratocysts (OK), adenomatoid odontogenic tumors (AOT) and solid ameloblastomas (AMBs). The sample was intentional and non-probabilistic, consisting of 20 cases of solid AMBs, 20 cases of AOT, 20 cases of OKs and 10 samples of DFs. The specimens were subjected to immunohistochemical method, using antibodies anti-Ing3, anti-Ing4, anti-Ing5 and antip53. Quantitative analysis was performed taking into account cytoplasmic and / or nuclear location for ING proteins and nuclear location for the p53 protein. The analysis of ING1 and ING2 immunoexpressions was performed in a previous study and the results were considered only for the correlation analysis. Data were analyzed by Statistical Package for Social Sciences for Windows (SPSS version 22.0; IBM, USA). Kruskal Wallis test was used to compare the immunoexpression between the groups of lesions, and Spearman test was used to investigate correlations. Values of p ≤ 0.05 were considered significant. This study showed a reduction in nuclear and cytoplasmic expression of ING3, ING4 and ING5 in odontogenic keratocysts (OKs) and ameloblastomas (AMBs). In addition, in some cases, loss of INGs nuclear expression was negatively correlated with p53 expression. Correlation analyzes may indicate existence of compensatory mechanisms between all the ING proteins in dental follicles (FDs) and adenomatoid odontogenic tumors (TOAs). These mechanisms seem to be less evident in COs and AMBs. The results of this study showed a reduction in ING3 expression in AMBs (p = 0.003); a reduction in ING4 expression, in OKs (p = 0.02) and in AMBs (p = 0.001); and a reduction in ING5 nuclear expression, also in OK (p = 0.09) and in AMBs (p = 0.012). Positive correlations were found between p53 nuclear expression with ING1 cytoplasm / nucleus expression (r = 0.603; p = 0.05) in OKs, and with ING3 cytoplasm / nucleus expression (r = 0.475; p = 0.034) and also ING4 cytoplasm / nucleus expression (r = 0.448; p = 0.047) in AMBs. Finally, this study suggests that reduction in the expression of INGs proteins seems to be an event that occurred in etiopathogenesis of more aggressive odontogenic lesions. Futhermore, nuclear / cytoplasmic expression of INGs proteins is not related to increase in p53 expression in OKs and AMBs, which indicates that loss of expression of these proteins may results in functional changes independently of p53 (AU).

Consideraciones Metodológicas en el Estudio Histomorfométrico del Hueso Interradicular de los Primeros Molares Inferiores de Ratas Wistar / Methodological Considerations in the Histomorphometric Study of the Interradicular Bone of the First Lower Molars of Wistar Rats

El objetivo fue evaluar el volumen óseo BV/TV (%) del hueso interradicular en ratas Wistar: A) en relación a la edad; B) en relación a la zona de estudio en animales de la misma edad. Se utilizó Grupo A) 15 ratas Wistar hembras de 6 (I), 10 (II) y 14 (III) semanas, Grupo B) 9 ratas Wistar macho de 8 semanas. Tras la eutanasia, se extrajeron los maxilares inferiores y se procesaron histológicamente para obtener cortes mesio-distales del primer molar inferior coloreados con H.E. Sobre microfotografías digitales se evaluó el BV/TV (%). El análisis estadístico se realizó en A) mediante ANOVA y Bonferroni test y en B) se calculó el rango (R). Los resultados en A) el BV/TV (%) aumenta significativamente con la edad de los animales; en B) se encontró que el BV/TV (%) varía hasta un 20% si se considera el volumen total y este rango disminuye a 8.3% al estudiar su mitad coronal. En conclusión, el BV/TV (%) del hueso interradicular del primer molar inferior de ratas Wistar varía considerablemente con la edad de los animales y en animales de una misma edad -según se considere evaluar todo el hueso interradicular del espacio alveolar o la mitad superior del mismo-. Los resultados de este trabajo recomiendan emplear animales de la misma edad y realizar mediciones histomorfométricas empleando la mitad coronal de dicho hueso, especialmente en los diseños de periodontitis experimental (AU)

Resin-modified glass ionomer containing calcium glycerophosphate: physico-mechanical properties and enamel demineralization

Abstract Sources of calcium and phosphate have been added to dental restorative materials to improve their anticaries effect. Objective This study evaluated the effect of adding calcium glycerophosphate (CaGP) to resin-modified glass ionomer cement (RMGIC) on the physico-mechanical properties, ion release, and enamel demineralization. Material and Methods: Specimens were fabricated for each experimental group: RMGIC without CaGP (Control), RMGIC with 1, 3 and 9% CaGP. To determine the release of fluoride (F), calcium (Ca) and phosphorus (P), six specimens were immersed in demineralization and remineralization solutions for 15 days. In another experimental trial, the following physico-mechanical properties were evaluated at time intervals of 1 and 7 days after fabrication: compressive strength (n=12), diametral tensile strength (n=12), surface hardness of material (n=6) and the degree of conversion of monomers (n=8). To study enamel demineralization, specimens (n=12) were attached to enamel blocks and submitted to pH-cycling. Subsequently, surface and cross-sectional hardness and the concentration of F, Ca and P in enamel were determined. Results The addition of CaGP to RMGIC led to higher mean release of F, Ca and P when compared with control (p<0.001). Mechanical properties were within the range of those of the ionomer cements after addition of 1% and 3% CaGP. The degree of conversion did not differ between groups at the 1st and the 7th day (p>0.439). The addition of 3% and 9% CaGP reduced mineral loss and increased F, Ca and P in the enamel when compared with control (p<0.05). Conclusion The addition of 3% CaGP in RMGIC increased the release of F, P and Ca, reduced enamel demineralization, and maintained the physico-mechanical properties within the parameters for this material.

Evaluation of dermal collagen stained with picrosirius red and examined under polarized light microscopy

Abstract: The special picrosirius red staining highlights the natural birefringence of collagen fibers when exposed to polarized light. The results from birefringence allow to evaluate the organization of the collagen fibers in the tissues. The authors intend to elucidate all steps to obtain and capture images of histological sections stained with picrosirius red and evaluated under polarized light microscopy, as well as possible artefacts that may occur.

Manejo Quirúrgico de Múltiples Condilomas de la Mucosa Oral en Paciente Bajo Terapia Antirretroviral de Gran Actividad / Surgical Management of Multiple Condylomas in the Oral Mucosa in Patient Undergoing Highly Active Antiretroviral Therapy

RESUMEN: El objetivo de este estudio fue reportar un caso clínico donde se realizó el manejo quirúrgico de múltiples condilomas de la mucosa oral en un paciente infectado por Virus de Inmunodeficiencia Humana (VIH) bajo Terapia Antirretroviral de Gran Actividad (TARGA). Hombre de 58 años en tratamiento por infección con VIH en TARGA hace 17 años, que acude al Servicio de Cirugía Maxilofacial del Hospital Barros Luco-Trudeau con múltiples lesiones verruciformes ubicadas en margen y cara dorsal de lengua, cara interna de ambas mejillas y labio inferior. Se realizó escisión quirúrgica de las lesiones de labio, cara dorsal de lengua y cara interna de mejilla del lado derecho, obteniéndose el diagnóstico histopatológico de condiloma. Tras 2 meses de realizar la cirugía se obtuvo recurrencia. La recurrencia de las lesiones puede originarse por la recrudescencia del virus latente adyacente al lecho quirúrgico y, por ello, deben considerarse otras alternativas de tratamiento. Por el impacto en la función, estética, potencial de contagio y malignización, es necesario su tratamiento.

ABSTRACT: The objective of this study was to report a clinical case in which surgical management of multiple condylomas in the oral mucosa was performed in a patient infected by Human Immunodeficiency Virus (HIV) who is under highly active antiretroviral therapy (HAART). A 58-yearold man, under HAART for 17 years for HIV infection, was admitted at the Maxillofacial Surgery Service at the Hospital Barros Luco-Trudeau as he was experiencing multiple verrucous lesions located on the lateral margin and dorsum of the tongue, as well as on the inner face of both cheeks and the lower lip. A surgical excision of the lesions on the lip, dorsum of the tongue and inner face of the right cheek was performed, where the histopathological diagnosis of condyloma was obtained. There was a recurrence two months after surgery. Recurrence of the lesions may be due to the recrudescence of the latent virus adjacent to the surgical bed and, therefore, other treatment alternatives should be considered. Treatment is necessary due to the impact on the function, aesthetics, and the potential to become contagious and malignant.

Histomorphometric analysis of the skin of women during the reproductive period

OBJECTIVES: The aim of this study was to evaluate the histomorphometry of the skin of women during the reproductive period according to the Fitzpatrick classification. METHODS: Thirty women aged 30 to 45 years were included in this study. We studied the surgical sites of extracted nevi. The material was processed for routine histology and then stained with haematoxylin and eosin as well as Picrosirius red. Four-micrometre histological sections were analysed according the Fitzpatrick criteria (skin pigmentation). The skin thickness and collagen concentration were determined for the reticular dermal skin. The data were statistically analysed with ANOVA. RESULTS: Fitzpatrick skin types I and II were thicker than the other skin types. CONCLUSIONS: Our data suggest that white skin may be less thick than dark skin.

Análise da expressão imunoistoquímica de regy e de proteínas envolvidas na regulação da apoptose em carcinomas epidermoides de língua oral / Analysis of the immunohistochemical expression of regy and of proteins involved in the regulation of apoptosis in oral tongue squamous cell carcinomas

O câncer na cavidade oral é uma das lesões malignas mais frequentes na população mundial. Como o processo de desenvolvimento das neoplasias malignas remete a danos que promovem um desequilíbrio na regulação da divisão e morte celular, nos últimos anos, diversos estudos foram realizados com o intuito de verificar a influência desses danos no comportamento global das células e na evolução da doença. Nesse contexto, pesquisas recentes mostraram que alterações na expressão de REG podem desempenhar um importante papel na progressão tumoral de várias neoplasias malignas, por interferir na regulação da apoptose. Diante disso, o objetivo desse trabalho foi investigar a expressão imunoistoquímica dos marcadores REG, p53, MDM-2, Bcl-2 e Bax em carcinomas epidermoides de língua (CELs) oral, com a finalidade de promover uma análise comparativa da imunoexpressão destas proteínas com os parâmetros clínico-patológicos de agressividade da lesão, no intuito de identificar se o REG contribui para a progressão do tumor e se interfere na expressão das proteínas relacionadas a apoptose. Para tanto, foram coletadas informações clínicas de 58 pacientes acometidos por CELs. Em seguida, foi realizada análise histopatológica e imunoistoquímica dos marcadores supracitados, em amostras de material biológico parafinado da lesão. Os resultados mostraram que os tumores com metástase nodal e de alto grau histopatológico de malignidade apresentavam percentuais significativamente menores de REG (p<0,05). Em adição, a análise da expressão de p53, MDM-2 e Bax nos diferentes parâmetros clínico-patológicos avaliados nesse trabalho, não revelou diferenças significativas nos percentuais de imunopositividade desses marcadores. Com relação ao Bcl-2, foi visto que tumores de alto grau de malignidade e com óbitos relacionados a doença apresentavam percentual significativamente menor de positividade dessa proteína (p<0,05). Por fim, o teste de correlação de Spearman demonstrou existir fraca correlação positiva, estatisticamente significativa, entre os percentuais de REG e das proteínas MDM-2 e Bcl-2. Baseado nesses achados, pode-se concluir que a redução da expressão de REG parece contribuir para a progressão do CEL oral e pode influenciar na expressão das proteínas relacionadas a regulação da apoptose (AU).

Cancer of the oral cavity is one of the most common malignancies in the world. Since the development of malignant neoplasms is related to damage that causes an imbalance in the regulation of cell division and cell death, many studies have been conducted in recent years to verify the influence of this damage on overall cell behavior and on the progression of the disease. Within this context, recent studies suggest that changes in the expression of REG play an important role in the progression of different malignant tumors by interfering with the regulation of apoptosis. Therefore, the objective of the present study was to investigate the immunohistochemical expression of REG, p53, MDM-2, Bcl-2 and Bax in oral tongue squamous cell carcinoma (OTSCC), correlating the immunoexpression of these proteins with clinicalpathological parameters of tumor aggressiveness, in order to determine whether REG contributes to tumor progression and interferes with the expression of apoptosisrelated proteins. For this purpose, the clinical data of 58 patients with OTSCC were collected and paraffin-embedded tumor specimens were submitted to histopathological analysis and immunohistochemistry using the markers cited above. The results showed significantly higher expression of REG (p<0.05) in low-grade tumors and without lymph node metastases. In addition, comparison of the expression patterns of p53, MDM-2 and Bax according to the clinical-pathological parameters studied revealed no significant differences in the percentage of immunostaining for these markers. Regarding Bcl-2, a significantly lower percentage of immunostaining for this protein was observed in tumors with a high grade of malignancy and tumors associated with deaths related to the disease (p<0.05). Finally, Spearman's correlation test demonstrated a significant weak positive correlation between the percentages of REG and of MDM-2 and Bcl-2. These findings suggest that the reduced expression of REG contributes to the progression of OTSCC and may influence the expression of proteins related to the regulation of apoptosis (AU).

Microglia and neurons in the hippocampus of migratory sandpipers

The semipalmated sandpiper Calidris pusilla and the spotted sandpiper Actitis macularia are long- and short-distance migrants, respectively. C. pusilla breeds in the sub-arctic and mid-arctic tundra of Canada and Alaska and winters on the north and east coasts of South America. A. macularia breeds in a broad distribution across most of North America from the treeline to the southern United States. It winters in the southern United States, and Central and South America. The autumn migration route of C. pusilla includes a non-stop flight over the Atlantic Ocean, whereas autumn route of A. macularia is largely over land. Because of this difference in their migratory paths and the visuo-spatial recognition tasks involved, we hypothesized that hippocampal volume and neuronal and glial numbers would differ between these two species. A. macularia did not differ from C. pusilla in the total number of hippocampal neurons, but the species had a larger hippocampal formation and more hippocampal microglia. It remains to be investigated whether these differences indicate interspecies differences or neural specializations associated with different strategies of orientation and navigation.

Morphometric analysis of feedforward pathways from the primary somatosensory area (S1) of rats

We used biotinylated dextran amine (BDA) to anterogradely label individual axons projecting from primary somatosensory cortex (S1) to four different cortical areas in rats. A major goal was to determine whether axon terminals in these target areas shared morphometric similarities based on the shape of individual terminal arbors and the density of two bouton types: en passant (Bp) and terminaux (Bt). Evidence from tridimensional reconstructions of isolated axon terminal fragments (n=111) did support a degree of morphological heterogeneity establishing two broad groups of axon terminals. Morphological parameters associated with the complexity of terminal arbors and the proportion of beaded Bp vs stalked Bt were found to differ significantly in these two groups following a discriminant function statistical analysis across axon fragments. Interestingly, both groups occurred in all four target areas, possibly consistent with a commonality of presynaptic processing of tactile information. These findings lay the ground for additional work aiming to investigate synaptic function at the single bouton level and see how this might be associated with emerging properties in postsynaptic targets.

Metástasis pulmonar de tumor de células de la granulosa como recurrencia del tumor primario después de diez años del tratamiento quirúrgico / Pulmonary metastasis of granulosa cell tumor as recurrence of primary tumor after 10 years of surgical treatment

Los tumores de células de la granulosa son neoplasias de bajo grado, que corresponden al 2-5% de los tumores malignos del ovario y presentan una baja incidencia. Las manifestaciones clínicas dependerán del tamaño tumoral y de la exposición prolongada a estrógenos producidos por las células neoplásicas. Habitualmente, son tumores unilaterales, sólido-quísticos con focos de hemorragia, constituidos por células pálidas con su característico pliegue nuclear, en un trasfondo fibrotecomatoso, con cuerpos de Call Exner sólo en el 30-60% de los casos. El diagnóstico se realiza con los niveles séricos de estradiol y con exámenes imagenológicos como ecotomografía ginecológica, tomografía axial computarizada o resonancia nuclear magnética. El tratamiento quirúrgico es la elección. Las recurrencias pueden ocurrir años posteriores al diagnóstico inicial y en general son pélvicas. El factor pronóstico más determinante en la evolución de la enfermedad es el estadio clínico al momento del diagnóstico. Se presenta el caso clínico de una paciente postmenopáusica con metástasis pulmonar de tumor de células de la granulosa después de diez años del tratamiento quirúrgico.

The granulosa cell tumors are low grade neoplasias that correspond to 2-5% of all malignant tumors of the ovary with a low incidence in the population. The clinical presentation depends on the tumor size and the long term exposition to estrogens produced by neoplastic cells. They are usually unilateral tumors exhibiting a mixture of cystic and solid areas with bleeding, constituted by pale cells with their characteristic nuclear groove in a fibrotecomatous background, with Call Exner bodies only in 30-60% of cases. The diagnosis is done with serum levels of estradiol and gynecological imaging analysis such as echotomography, Computerized axial tomography and magnetic nuclear resonance. The surgical treatment is the choice. The recurrences could happen years after the first diagnosis and usually are in pelvic area. The most important prognostic factor in the disease progression is the clinical stage at diagnosis. It is presented a case report of a postmenopausal patient with lung metastasis of granulosa cell tumor after ten years of surgical treatment.

Alteraciones anatómicas en epidermis de hojas infectadas de Phoradendron bathyoryctum Eichler (Viscaceae) / Anatomical alterations in epidermis of infected leaves of Phoradendron bathyoryctum Eichler (Viscaceae)

Phoradendron bathyoryctum Eichler (Viscaceae) es una planta hemiparásita que crece en la República Argentina sobre varias especies arbóreas. Se utiliza en medicina popular de la misma manera que el muérdago europeo (Viscum album L. ­Viscaceae­), para disminuir la presión arterial. Los muérdagos, a veces, pueden ser susceptibles de epiparasitismo, o también sufrir el ataque de diferentes agentes fitopatógenos. El objetivo de este estudio es analizar los cambios anatómicos que se producen en las hojas de Ph. bathyoryctum cuando son afectadas por algún agente infeccioso. Se colectaron ejemplares sanos e infectados, los últimos, con lesiones foliares superficiales. Se realizaron técnicas de raspado, diafanizado y obtención de epidermis en las hojas de los ejemplares. También se practicaron reacciones histoquímicas. En las hojas infectadas se observó una destrucción progresiva de los estomas y cambios en el resto de los tejidos epidérmico y subepidérmico, que dieron como resultado la formación de estructuras visibles macroscópicamente, y semejantes a lenticelas foliares. De acuerdo con las observaciones realizadas y por comparación con la bibliografía consultada, se supone que el daño foliar se inicia en el estoma donde se producen cambios estructurales hasta su destrucción, la modificación de las células adyacentes y la formación de lesiones superficiales en las hojas infectadas. En cuanto al agente, si bien no se pudo determinar en esta oportunidad, podría asociarse a algún tipo de virus. Se sugiere realizar futuros estudios para confirmar si los virus son los causantes de las lesiones. (AU)

Ultrastructure of the surface of dental enamel with molar incisor hypomineralization (MIH) with and without acid etching / Ultraestructura de la superficie del esmalte dental con hipomineralización molar incisiva (mih) con y sin grabado ácido

El objetivo del presente trabajo fue analizar la ultraestructura y composición mineral de la superficie del esmalte de un molar con MIH, con y sin tratamiento de grabado ácido. Se analizaron una pieza dentaria permanente sin lesiones clínicas de MIH (controles) y una pieza con diagnóstico clínico de MIH leve y moderada, con indicación de extracción con y sin grabado acido (H3PO4 37 por ciento, 20”). Fueron procesadas para su observación con microscopio electrónico de barrido (SEM) ZEISS (Supra 40) y análisis de la composición mineral con detector EDS (Oxford Instruments). El esmalte del control mostró superficie prismática y patrón de grabado normales. El esmalte clínicamente sano en la pieza con MIH reveló una pérdida parcial del patrón prismático. La lesión leve se presentó porosa con ocasional esgrietas. La moderada presentó mayor porosidad, con grietas de mayor tamaño y presencia de gran cantidad de escamas. La composición mineral de las superficies afectadas mostró menor contenido de Ca y P y aumento de O y C. En la muestra con MIH, inclusive con aspecto normal, las desmineralizaciones no responden a un patrón de grabado, mostrando exposición de cristales con redondeamiento en los extremos de las varillas y menor desmineralización en los espacios interprismáticos. El grabado ácido incrementó la aparición de grietas y profundos poros en la estructura adamantina del esmalte con lesión. La composición mineral mostró en las lesiones moderadas una mayor disminución del contenido de Ca, P y Cl.El esmalte con MIH, inclusive en la superficie adamantina clínicamente intacta, presenta severas alteraciones ultraestructurales y cambios en la composición iónica afectando el patrónde grabado ácido, que podría interferir con los mecanismos de adhesión.

The aim of the present work was to analyze the ultrastructure and mineral composition of the surface of the enamel on amolar with MIH, with and without acid etching. A permanent tooth without clinical MIH lesions (control) and a tooth with clinical diagnosis of mild and moderate MIH, with indication for extraction, were processed with and without acid etching(H3PO437%, 20”) for observation with scanning electron microscope (SEM) ZEISS (Supra 40) and mineral composition analysis with an EDS detector (Oxford Instruments). The control enamel showed normal prismatic surface and etching pattern. The clinically health y enamel on the tooth with MIH revealed partial loss of prismatic pattern. The mild lesion wasporous with occasional cracks. The moderate lesion was moreporous, with larger cracks and many scales. The mineral composition of the affected surfaces had lower Ca and Pcontent and higher O and C. On the tooth with MIH, even on normal looking enamel, the demineralization does not correspond to an etching pattern, and exhibits exposure of crystals with rods with rounded ends and less demineralization in the inter-prismatic spaces. Acid etching increased the presence of cracks and deep pores in the adamantine structure of the enamel with lesion. In moderate lesions, the mineral composition had higher content of Ca, P and Cl. Enamel with MIH, even on clinically intact adamantine surfaces, shows severe alterations in the ultrastructure and changes in ionic composition, which affect the acid etching pattern and may interfere with adhesion.

Pulpectomía lateral: una técnica de micro-odontología para desarrollar habilidad en el manejo de microscopios dentales / Lateral pulpectomy: a microtechnique to develop competency in the use of the dental microscope

Con la aparición de cada nuevo sistema de diagnóstico o tratamiento dental, es necesario capacitar a los cirujanos dentistas para su correctoempleo; los microscopios dentales no escapan a esta regla. En las últimasdos décadas el uso de microscopios dentales se ha incrementado, pero aún no se establece una reglamentación respecto a su empleo y son muy pocos los profesionistas que cuentan con estos equipos para su práctica privada, es por ello que no se han estandarizado técnicas para que los clínicos desarrollen las habilidades necesarias que les permitan utilizar los equipos de microscopia en su práctica cotidiana en forma segura y con facilidad. La técnica de ®pulpectomía lateral¼ es un ejerciciode micro-odontología que se realiza en dientes extraídos, que no tiene aplicación clínica como tal, pero es parte de una serie de procedimientos que tienen la fi nalidad de desarrollar las habilidades de orientación, movimientoy aplicación de presión. Estas habilidades son indispensablescuando se trabaja en la boca de los pacientes con microscopios dentales

As new diagnostic and dental treatments emerge, dental surgeons need to be trained in their correct use. Dental microscopes are no exception to this rule. In the last two decades, the use of dental microscopes has increased, though no set rules for their use have been defi ned. Moreover, few professionals have this equipment in their private practice, which is why techniques have not been standardized for clinicians to develop the skills required for them to be able to use dental microscopes easily and safely in their daily practice. The ®lateral pulpectomy¼ technique is a microdentistry training exercise that is performed on extracted teeth. Though it has no clinical application as such, it forms part of a series of procedures that are designed to develop directional, motion, and pressure skills, all of which are essential when working with dental microscopes in the mouths of patients.

Activación sónica versus ultrasónica de EDTA al 10% para remoción de barrillo dentinario en el tercio apical del canal radicular / Sonic versus ultrasonic activation of 10% EDTA for smear layer removal in the root canal apical third

El objetivo de este estudio fue evaluar la efectividad de la activación sónica y ultrasónica del ácido etilendiaminotetraacético (EDTA) al 10% para remover barrillo dentinario en el tercio apical y el grado de erosión producido sobre las paredes del canal radicular. Se realizó un estudio cuantitativo experimental in vitro. Se instrumentaron 40 dientes manualmente con la técnica step-back hasta lima maestra número 55 y fueron asignados a 4 grupos según protocolo de irrigación final: I: agua destilada estéril sin activación, II: EDTA 10% sin activación por 30 segundos, III: EDTA 10% activado sónicamente por 30 segundos y IV: EDTA 10% activado ultrasónicamente por 30 segundos. Las muestras fueron observadas en MEB a una magnificación de 5000X. Se evaluó la presencia de barrillo dentinario remanente y grado de erosión según criterios de Torabinejad et al. Para el análisis estadístico se utilizó la prueba de Kruskal-Wallis y Post Hoc de Tukey con un nivel de significancia 5% (p=0,05). El grupo I (agua sin activación) presentó mayor cantidad de barrillo que el resto de los grupos (p<0,01). En cuanto a la presencia de barrillo, entre los grupos II (EDTA sin activación), III (EDTA con sonido) y IV (EDTA con ultrasonido) no hubo diferencias estadísticamente significativas. En cuanto a la erosión, el grupo II presentó una mayor erosión que el resto siendo significativa con los grupos I y III (p=0,006). Entre los grupos I, III y IV no hubo diferencias estadísticamente significativas. La activación sónica y ultrasónica del EDTA 10% no produjo una remoción de barrillo significativamente superior al compararlo con la irrigación convencional, pero sí hubo diferencias en cuanto a erosión, siendo menor con la activación sónica

The purpose of this study was to observe the effect of passive sonic and ultrasonic irrigation with 10% Ethilenediaminotetracethic acid (EDTA) on smear layer of the apical third of instrumented root canals as well as the level of erosion produced on the canal walls. For this quantitative experimental in vitro study 40 teeth were hand instrumented with step back technique until master file num 55 and divided in four groups according to the final irrigation protocol: I: distilled water without activation, II: 10% EDTA for 30 seconds without activation, III: 10% EDTA sonic activated for 30 seconds, IV: 10% EDTA ultrasonic activated for 30 seconds. Samples were prepared for MEB observation at 5000X. The presence of smear layer and erosion on dentine walls was quantified according to Torabinejad et al. criteria. Data were submitted at Kruskal-Wallis test and Tukey Post Hoc with a significance level of 5% (P=0.05). When evaluating the smear layer presence, there were no statistically significant differences between the groups II, III and IV. Group I had a highest amount of smear layer (p<0.01). As for erosion, group II showed higher erosion than the others, being significant with groups I and III (p=0.006). Among groups I, III and IV there were not statistically significant differences. Sonic and ultrasonic activation of EDTA 10% did not produce a significantly superior removal of smear layer when compared with conventional irrigation. Erosion of root canal walls was significantly lower after sonic activation