Comparación Morfométrica de Esqueletos Transparentados de Cuatro Especies de Roedores Myomorfos / Morphometric Comparison of Transparent Skeletons of Four Species of Myomorphic Rodents

Las técnicas de doble tinción y transparentación se han usado desde 1897, pero su utilidad ha sido poco explorada en los estudios anatómicos de micromamíferos adultos. No obstante, la combinación de estas técnicas con el análisis alométrico mutivariado posibilita el estudio de esqueletos poscraneales articulados de tales grupos de micromamíferos como los roedores, los cuales son muy limitados ya que casi siempre se enfocan en los cráneos. En este estudio, analizamos y comparamos la morfometría del esqueleto de Neotomodon alstoni con la de Meriones unguiculatus, Phodopus campbelli y Rattus norvegicus. Usamos la técnica de doble tinción y transparentación para analizar las relaciones morfométricas entre estos roedores utilizando sesenta caracteres esqueléticos. Se encontró que tres especies comparten dos correlaciones comunes y compartieron el mismo tipo de crecimiento isométrico en una de ellas; además se encontraron similitudes aparentes entre los patrones de la morfometría de P campbelli con el patrón de osificación descrito para la especie relacionada Mesocricetus auratus. Las diferencias en el crecimiento alométrico pueden representar también diferencias en el ritmo de desarrollo de acuerdo con el tipo de historia de vida de cada especie. Aquí demostramos que tanto la técnica de preparación como el método de análisis morfométricos son herramientas poderosas pero simples, para realizar estudios anatómicos y morfológicos en el laboratorio. Nuestros resultados reflejan las condiciones del desarrollo ontogenético derivados delpropio patrón de heterocronía para cada especie, y además representan la historia evolutiva de este grupo analizado. Sin embargo, consideramos que es deseable más investigación.

SUMMARY: Clearing and staining techniques have been present since 1897, However, their use in anatomical studies of adult micromammals has been limited. When using such techniques in combination with allometric method, it is possible to study articulated skeletons of micromammals, instead of relying only on the skulls, which is important in morphologically complicated groups as the rodents. Research involving multivariate allometric analysis of postcranial skeleton of rodents has been limited and confined to specific items. In this study, we analyzed and compared the morphometry of the skeleton of Neotomodon alstoni with that of Meriones unguiculatus, Phodopus campbelli and Rattus norvegicus. We applied the double staining and clearing technique in order to determine the morphometric relation between these rodents using sixty skeletal characters. We found that three species share two common correlations and one isometric, with apparent similarities between the morphometry patterns of P campbelli with the ossification pattern described for the related species Mesocricetus auratus. The differences in allometric growth could represent differences in the development stages according to the type of life history for each species. In this analysis we confirmed that both the preparation technique and morphometric analysis method, are simple yet verifiable tools for anatomical and morphological studies. Our results reflect the conditions of ontogenetic development derived from the heterochrony pattern for each species, representing the evolutionary history for this group. Therefore, as this approach continues to be discussed, ongoing research is warranted.

Consistency comparison of programmed cell death 1-ligand 1 in different immuno-histochemical staining methods / 北京大学学报(医学版)

OBJECTIVE@#To compare the consistency of programmed cell death 1-ligand 1 (PD-L1, clone E1L3N, 22C3, SP263) in different immunohistochemical staining methods.@*METHODS@#The first step was to select the optimal process: The PD-L1(clone E1L3N) antibody recommended process, self-built process ①, self-built process ② and self-built process ③ were used to perform immunohistochemical staining in 5 cases of tonsil tissue. The quality of all slides was scored by expert pathologists (0-6 points). The process with the highest score was selected. The second step was to compare the consistency between the optimal procedure and the two standard procedures. Thirty-two cases of lung non-small cell carcinoma diagnosed by pathology in Peking University First Hospital in the past two years were randomly selected. The 32 cases were stained in parallel with the SP263 and 22C3 standard procedures, and all stained slides were scored by specialized pathologists for tumor proportion score (TPS). The scoring results were grouped according to < 1%, ≥1% to < 10%, ≥10% to < 50%, and ≥50%. The consistency of PD-L1 detection antibody clone E1L3N and 22C3, E1L3N and SP263 staining results was analyzed.@*RESULTS@#Tonsil stained slides scores (0-6 points) were as follows: The recommended protocol was 5, 5, 5, 5 and 5. The self-built process ① was 5, 6, 6, 5 and 6. The self-built process ② was 4, 4, 4, 4 and 4.The self-built process ③ was 3, 3, 3, 3 and 3. The self-built process ① was the best with the highest score. The TPSs of 32 non small cell lung carcinoma (NSCLC) cases were as follows: Of self-built process ①, 6 cases were lower than 1%, 5 cases were from 1% to 10%, 10 cases were from 10% to 50%, and 11 cases were higher than 50%; of 22C3 standard procedure, 5 cases were lower than 1%, 3 cases were from 1% to 10%, 13 cases were from 10% to 50%, 11 cases were higher than 50%; of SP263 standard procedure, 7 cases were lower than 1%, 4 cases were from 1% to 10%, 11 cases were from 10% to 50%, 10 cases were higher than 50%. The results of the consistency test were as follows: The κ value for self-built process ① and 22C3 standard procedure was 0.736 (P < 0.001), the agreement was good; the κ value for self-built process ① and SP263 standard procedure was 0.914 (P < 0.001), the agreement was very good.@*CONCLUSION@#The immunostaining using PD-L1(E1L3N) with validated self-built staining protocol ① by Ventana Benchmark GX platform can obtain high quality of slides, and the TPSs based on these slides are in good agreement with 22C3 and SP263 standard procedures.

Effects of black tea tooth staining previously to 35% hydrogen peroxide bleaching

Aim: To determine if the artificial staining with black tea (BT) influences the enamel microhardness before in-office bleaching and if BT staining is necessary to evaluate the efficacy of bleaching with 35% hydrogen peroxide Methods: Enamel/dentin blocks were randomized into groups according to the staining protocol (n=5/group): (CO) control ­ maintained in artificial saliva solution (AS); (BT4) immersed in black tea solution for 4 h; (BT24) immersed in black tea solution for 24 h. After the staining protocols, all specimens were kept in AS for one week, followed by bleaching (three sessions of HP application for 40 min). Knoop surface microhardness (kgF/mm2) was determined at baseline (T0), after staining (T1), after 7 days of storage in AS (T2), and after bleaching (T3). The color (∆E00) and coordinate changes (∆L, ∆a, ∆b) were measured using a digital spectrophotometer at T0 and T3. Data were submitted to one-way (∆E00, ∆L, ∆a, ∆b) or two-way ANOVA repeated measures (kgF/mm2) and Tukey's test (a=5%). Results: The staining protocols (BT4 and BT24) promoted significantly lower microhardness (T1 and T2, p<0.05) than CO, whereas CO was the only group to maintain microhardness values over time. Bleaching promoted perceptible ∆E00 without a significant difference among the groups regardless of the staining protocol (p=0.122). CO and BT4 showed no differences in terms of ∆L and ∆a (p>0.05), but BT4 displayed a higher ∆b than CO. Conclusion:The artificial staining with BT negatively affected the enamel surface microhardness and was not essential to evaluate the efficacy of 35% hydrogen peroxide bleaching

Uso de la pasta CDZ en dientes temporarios necrosados con una técnica mínimamente invasiva / Use of CDZ paste in necrotic primary teeth with a minimally invasive technique

Introducción: El tratamiento de los canales radiculares de molares temporales con pulpas infectadas ha sido ampliamente descrito y motivo de discusión por muchos años, no existiendo aún un consenso en cuanto al material de obturación. La pasta CTZ (cloranfenicol, tetraciclina y óxido de zinc más eugenol) acompañada de la Técnica de Endodoncia No Instrumentada ha mostrado una alta efectividad clínica y radiográfica para el tratamiento de molares temporales con compromiso pulpar. Se ha propuesto el reemplazo del componente Tetraciclina por Doxiciclina de la formulación, por las implicancias de un posible amelo-génesis imperfecta y la coloración en la corona de dicho componente. Objetivo. Evaluar la efectividad clínica de la pasta CDZ en el tratamiento endodóntico de dientes primarios necrosados con una técnica mínimamente invasiva. Metodología. Estudio de intervención en el que se incluyeron pacientes que presentaban dientes temporales con indicación de terapia pulpar y en quienes se utilizó la pasta CDZ. El éxito del tratamiento se midió por la desaparición de la sintomatología. Resultados. Se incluyeron en el estudio 76 pacientes entre 2 a 9 años. La eficacia del tratamiento con CDZ fue del 97,6% en 125 dientes. Conclusiones. Los hallazgos son comparables a los estudios que utilizaron el CTZ. Debido a las características biológicas del material, su bajo costo, fácil manipulación y excelentes resultados clínicos, se considera una opción en la terapia pulpar en dientes temporales, como una alternativa en el uso de programas de salud pública.

Introduction: Treatment of root canals of primary molars with infected pulps has been widely described and has been the subject of discussion for many years, and there is still no consensus regarding the filling material. The CTZ paste (chloramphenicol, tetracycline and zinc oxide plus eugenol) accompanied by the Non-Instrumented Endodontic Technique has shown high clinical and radiographic effectiveness for the treatment of primary molars with pulp involvement. The replacement of the Tetracycline component by Doxycycline of the formulation has been proposed, due to the implications of a possible amelogenesis imperfecta and the coloration in the crown of said component. Goal. To evaluate the clinical effectiveness of CDZ paste in the endodontic treatment of necrotic primary teeth with a minimally invasive technique. Methodology: Intervention study in which patients with temporary teeth with an indication for pulp therapy and in whom CDZ paste was used were included. Treatment success was measured by the disappearance of symptoms. Results. 76 patients between 2 and 9 years old were included in the study. The efficacy of CDZ treatment was 97.6% in 125 teeth. Conclusions: The findings are comparable to studies using CTZ. Due to the biological characteristics of the material, its low cost, easy handling and excellent clinical results, it is considered an option in pulp therapy in primary teeth, as an alternative in the use of public health programs.

An experimental study of tangzhouling on morphological changes of nissl bodies in the dorsal root ganglion of DM rats / Un estudio experimental de tangzhouling sobre los cambios morfológicos de los cuerpos de nissl en el ganglio de la raíz dorsal de ratas DM

SUMMARY: This study aims to investigate the effect of Tangzhouling on the morphological changes of Nissl bodies in the dorsal root ganglion of DM Rats. In this study, 69 rats were randomly divided into a control group (n = 10) and a model group (n = 59). The rats in the model group were randomly divided into a diabetic group (n = 11), a vitamin C group (n = 12), a low dose Tangzhouling group (n = 12), a medium dose Tangzhouling group (n = 12) and a high dose Tangzhouling group (n = 12). The dose of Tangzhouling in the low dose group was 5 times that of the adult dose, being 0.44g/kg/d. The dose of Tangzhouling in the medium dose group was 10 times that of the adult dose, being 0.88g/kg/d. The dose of Tangzhouling in the high dose group was 20 times that of the adult dose, being 1.75g/kg/d. All doses above are crude drug dosages. Rats in the vitamin C group were given 10 times the dose of an adult, being, 0.05 g/ kg/d. The diabetic group and the control group were given the same amount of distilled water. Drug delivery time is 16 weeks. The dorsal root ganglion was placed in a freezing tube at the end of the experiment. The morphological changes of Nissl bodies in the dorsal root ganglion were detected by HE and Nissl staining. The study results showed that vitamin C had no significant effect on the quantity, size and nucleolus. Tangzhouling can improvee the morphology, quantity and nucleolus of Nissl bodies to a certain extent, and the high dose is better than the lower dose. Tangzhouling capsules can improve the nerve function of DM rats through Nissl bodies.

RESUMEN: Este estudio tuvo como objetivo investigar el efecto de Tangzhouling en los cambios morfológicos de los cuerpos de Nissl en el ganglio de la raíz dorsal de las ratas DM. En este estudio, 69 ratas se dividieron aleatoriamente en un grupo control (n = 10) y un grupo modelo (n = 59). Las ratas del grupo modelo se dividieron aleatoriamente en un grupo diabéticos (n = 11), un grupo vitamina C (n = 12), un grupo de dosis baja de Tangzhouling (n = 12), un grupo de dosis media de Tangzhouling (n = 12) y un grupo de dosis alta de Tangzhouling (n = 12). La dosis de Tangzhouling en el grupo de dosis baja fue 5 veces mayor que la dosis del adulto, siendo 0,44 g/kg/d. La dosis de Tangzhouling en el grupo de dosis media fue 10 veces mayor que la dosis del adulto, siendo 0,88 g/kg/d. La dosis de Tangzhouling en el grupo de dosis alta fue 20 veces mayor que la dosis del adulto, siendo 1,75 g/kg/d. Todas las dosis anteriores son dosis de fármaco crudo. Se les administró 10 veces la dosis de un adulto a las ratas del grupo vitamina C, siendo 0,05 g/kg/d. El grupo de diabéticos y el grupo de control recibieron la misma cantidad de agua destilada. El tiempo de entrega del fármaco fue de 16 semanas. El ganglio de la raíz dorsal se colocó en un tubo de congelación al final del experimento. Los cambios morfológicos de los cuerpos de Nissl en el ganglio de la raíz dorsal se detectaron mediante tinción de HE y Nissl. Los resultados del estudio mostraron que la vitamina C no tuvo un efecto significativo sobre la cantidad, el tamaño y el nucléolo. Tangzhouling puede mejorar la morfología, la cantidad y el nucléolo de los cuerpos de Nissl hasta cierto punto, y es mejor la dosis alta que la dosis baja. Las cápsulas de Tangzhouling pueden mejorar la función nerviosa de las ratas DM a través de los cuerpos de Nissl.

Investigation of teratogenic effects of letrozole on fetal bone development / Investigación de los efectos teratogénicos de letrozol en el desarrollo óseo fetal

SUMMARY: Letrozole is mainly used for the treatment of unexplained infertility, breast cancer and polycystic ovarian syndrome, with secondary use in ovarian stimulation. In cases of unexpected or unknown pregnancy during the use of letrozole, letrozole may cause a teratogenic effect on the fetus. In this reason, in this study, we aimed to determine the effect of letrozole on fetal bone development. In this study, 32 pregnant Wistar albino rats were used. The rats were divided into four groups: Control (saline) and high; 0.3 mg/kg, medium; 0.03 mg/kg, low; 0.003 mg/ kg letrozole. Saline and letrozole were administered in 100 mL solutions by intraperitonaly from day 11 to day 15 of pregnancy. The skeletal system development of fetuses was examined with double skeletal staining, immunohistochemical staining methods and mineral density scanning electron microscopy. A total of 100 fetuses from female rats, 25 in each group, were included in the study. As a result of that, ossification rates were observed to decrease depending on the dose of letrozole in the forelimb limb (scapula, humerus, radius, ulna) and hindlimb (femur, tibia, fibula) limb bones. As a result of the statistical analysis, a statistically significant decrease was found in the ossification rates of all bones between the control group and low, medium, high letrozole groups (p<0.001). Exposure to letrozole during pregnancy adversely affected ossification and bone growth. However, the teratogenic effects of letrozole are unclear. Therefore, it needs to be investigated more extensively.

RESUMEN: Letrozol se usa principalmente para el tratamiento de la infertilidad inexplicable, el cáncer de mama y el síndrome de ovario poliquístico, con estimulación ovárica de uso secundario. En casos de embarazo inesperado o desconocido durante el uso de letrozol, puede causar un efecto teratogénico en el feto. Por esta razón, en este estudio, nuestro objetivo fue determinar el efecto de letrozol en el desarrollo óseo fetal. Se utilizaron 32 ratas albinas Wistar preñadas las cuales se distribuyeron en cuatro grupos: Control (solución salina) y alta; 0,3 mg/kg, medio; 0,03 mg/kg, bajo; 0,003 mg/kg de letrozol. Se administró solución salina y letrozol en soluciones de 100 mL por vía intraperitoneal desde el día 11 hasta el día 15 de la preñez. El desarrollo del sistema esquelético de los fetos se examinó con tinción esquelética doble, métodos de tinción inmunohistoquímica y microscopía electrónica de barrido de densidad mineral. Se incluyeron en el estudio un total de 100 fetos de ratas hembra, 25 en cada grupo. Como resultado, se observó que las tasas de osificación disminuían dependiendo de la dosis de letrozol en los huesos de los miembros torácicos (escápula, húmero, radio, ulna) y de las miembros pélvicos (fémur, tibia, fíbula). Se encontró una disminución estadísticamente significativa en las tasas de osificación de todos los huesos entre el grupo control y los grupos de letrozol bajo, medio y alto (p<0,001). La exposición a letrozol durante la preñez afectó negativamente la osificación y el crecimiento óseo. Sin embargo, los efectos teratogénicos del letrozol no están claros por lo que debe ser investigado más extensamente.

Produção de biscoitos recheados com formação de organogel na base gordurosa / Production of sandwich cookies with the formation of organogel in the fatty base

A fim de atender à demanda do público que atualmente busca por alimentos mais saudáveis, as indústrias têm procurado alternativas que possibilitem a aplicação de ingredientes que agreguem valor nutricional aos produtos. A redução de gorduras saturadas e trans em produtos alimentícios, bem como a inserção de cereais ou farinhas nutricionais, vem sendo aplicadas em produtos de panificação. Biscoitos recheados possuem como bases geralmente biscoitos à base de farinha de trigo. O objetivo foi desenvolver formulação de biscoitos recheados com substituição de gordura vegetal por organogel no recheio e de farinha de trigo por farinha de sorgo no biscoito, a fim de agregar valor nutricional ao produto. Foram desenvolvidos biscoitos recheados: 1) recheio controle e com substituição da gordura vegetal dos recheios por organogel elaborado com sistema emulsionado (colágeno + óleo vegetal + água), a fim de diminuir concentrações de gorduras saturadas e trans. 2) para a base elaborouse biscoitos controle (farinha de trigo) e com substituição parcial e total de farinha de trigo por farinha de sorgo em 50% (50FS) e 100% (100FS). Foram conduzidas nos recheios e das bases dos biscoitos análises físicas e físico-químicas (textura, atividade de água, cor, composição centesimal e reologia) para avaliação e para análise de estabilidade de 6 semanas. Os resultados apresentaram que o biscoito 50FS obteve melhor valor de textura (Controle: 16,09 ± 1,28 N; 50FS: 19,63 ± 5,68 N e 100FS: 10,09 ± 0,65 N) e menor teor de atividade de água (Semana 01: 0,327±0,01 e Semana 06: 0,389 ± 0,00) do que o biscoito controle, durante análise de estabilidade. O biscoito 100FS apresentou coloração mais avermelhada. Os biscoitos 50FS e 100FS apresentaram maior teor proteico do que o controle (Controle: 5,37 ± 0,23 %; 50FS: 5,64 ± 0,49 % e 100FS: 5,75 ± 0,49 %). O recheio com organogel apresentou maior dureza (N) durante análise de estabilidade do que o recheio controle (Semana 6 Organogel: 6,81±1,48; Controle: 4,29±0,38). Os parâmetros de adesividade, coesividade e gomosidade do recheio com organogel não apresentaram diferenças significativas (p > 0,05). Os valores de atividade de água da formulação com organogel foram mais altos do que o recheio controle (Semana 6 Organogel: 0,730±0,00; Controle: 0,555±0,01). O valor de L* foi maior para o recheio controle, apresentando coloração mais amarelada do que a formulação com organogel. O recheio com organogel apresentou redução de 65 % do teor lipídico e aumento do teor proteico. Os recheios controle, com organogel e de mercado apresentaram comportamento tixotrópico durante a avaliação reológica, sendo que o produto de mercado teve comportamento próximo à formulação controle, com recuperação quase total da estrutura. Foram desenvolvidos cinco produtos, sendo três inovadores com valor nutricional agregado, atendendo às legislações vigentes, vida útil mínima de 6 semanas e ao apelo do mercado atual, podendo ser comercializados como biscoito recheado

In order to satisfy the demand of the public that is currently looking for healthier foods industries have been looking for alternatives that allow the application of ingredients that add nutritional value to the products. The reduction of saturated and trans fats in food products, as well as the insertion of cereals or nutritional flours, has been applied in bakery products. Filled cookies are usually based on wheat flour. The objective was to develop a formulation of filled cookies with replacement of vegetable fat for organogel in the filling and wheat flour for sorghum flour in the biscuit, in order to add nutritional value to the product. In this study, cookies filled with vegetable fat and wheat flour were used as a control where: 1) filling was replaced by organogel elaborated with an emulsified system (collagen + vegetable oil + water); and 2) base was prepared with partial and total replacer of wheat flour for sorghum flour in 50% (50FS) and 100% (100FS). Physical and physicochemical analyzes (texture, water activity, color, proximate composition and rheology) were carried out on the fillings and bases of the biscuits for evaluation and for the stability analysis of 6 weeks. The results showed that the 50FS cookies had a better texture value (Control: 16,09±1,28 N; 50FS: 19,63±5,68N and 10,09±0,65 N) and lower content of water activity (Week 1: 0,327±0,01 and Week 6: 0,389±0,00) than the control cookie during stability analysis. The 100FS had a more reddish color. The 50FS and 100FS cookies had a higher protein content than the control (Control: 5,37±0,23 %; 50FS 5,64±0,49 %). The fillings with organogel showed a higher hardness (N) than the control during stability analysis (Week 6 Organogel: 6,81±1,48; Control: 4,29±0,38). The parameters of adhesiveness, cohesiveness and guminess of the filling with organogel showed no significant differences (p> 0.05). The water activity values of the organogel formulation were higher than the control filling (Week 6 Organogel: 0,730±0,00; Control: 0,555±0,01). The value of L * was higher for the control filling, showing a more yellowish color than the formulation with organogel. The filling with organogel showed a 65% reduction in lipid content and an increase in protein content. The control, organogel and market fillings showed a thixotropic behavior in the rheological evaluation, and the market product had a behavior close to the control formulation, with almost total recovery of the structure. Five products were developed, three of which were innovative with added nutritional value, in compliance with current legislation, a minimum shelf life of 6 weeks, which can be sold as a stuffed cookies.

Identification of new human mesenchymal stem cell biomarker by aptamers / Identificação de novos biomarcadores de células tronco mesenquimais de origem humana por meio de aptâmeros

Stem cells are undifferentiated cells that can be distinguished from others by their ability to self-renew and to differentiate into new specific cell types. Mesenchymal stem cells (MSC) are adult stem cells that can be obtained from different sources, such as adipose tissue, bone marrow, dental pulp, and umbilical cord. They can either replicate, originating new identical cells, or differentiate into cells of mesodermal origin and from other germ layers. MSC have been studied as new tools for regenerative therapy. Although encouraging results have been demonstrated, MSC-based therapies still face a great barrier: the difficulty of isolating these cells from heterogeneous environments. MSC are currently characterized by immunolabelling through a set of multiple surface membrane markers, including CD29, CD73, CD90 and CD105, which are also expressed by other cell types. Hence, the present work aimed to identify new specific biomarkers for the characterization of human MSC using DNA aptamers produced by the SELEX (Systematic Evolution of Ligands by EXponential Enrichment) technique. Our results showed that MSC from different origins bound to DNA candidate aptamers, that is, DNA or RNA oligonucleotides selected from random libraries that bind specifically to biological targets. Aptamer-bound MSC could be isolated by fluorescenceactivated cell sorting (FACS) procedures, enhancing the induction of differentiation into specific phenotypes (chondrocytes, osteocytes and adipocytes) when compared to the whole MSC population. Flow cytometry analyses revealed that candidate aptamers bound to 50% of the MSC population from dental pulp and did not present significant binding rates to human fibroblasts or lymphocytes, both used as negative control. Moreover, immunofluorescence images and confocal analyses revealed staining of MSC by aptamers localized in the surfacemembrane of these cells. The results also showed internal staining of human monocytes by our investigated aptamers. A non-specific control aptamer (CNTR APT) obtained from the random pool was then utilized to compare the specificity of the aptamers bound to the analyzed non-apoptotic cells, showing no staining for MSC. However, 40% of the monocytes bound to the CNTR APT. Normalized data based on the cells bound to candidate aptamers compared to those bound to the CNTR APT, revealed a 10 to 16-fold higher binding rate for MSC against 2-fold for monocytes. Despite its low specificity, monocyte-aptamer binding occurs probably due to the expression of shared markers with MSC, since monocytes are derived from hematopoietic stem cells and are important for the immune system ability to internalize/phagocyte external molecules. Given that, we performed a pull-down assay followed by mass spectrometry analysis to detect which MSC-specific protein or other target epitope not coexpressed by monocytes or the CNTR APT would bind to the candidate aptamer. Distinguishing between MSC and monocyte epitopes is important, as both cells are involved in immunomodulatory effects after MSC transplantations. ADAM17 was found to be a target of the APT10, emerging as a possible biomarker of MSC, since its involvement in the inhibition of the TGF signaling cascade, which is responsible for the differentiation of MSC. Thus, MSC with a higher stemness profile should overexpress the protein ADAM17, which presents a catalytic site with affinity to APT10. Another target of Apt 10 is VAMP3, belonging to a transmembrane protein complex that is involved in endocytosis and exocytosis processes during immune and inflammatory responses. Overall, proteins identified as targets of APT10 may be cell surface MSC biomarkers, with importance for MSC-based cell and immune therapies

Células tronco são células indiferenciadas que podem ser distinguidas de outros tipos celulares por meio da habilidade de se auto renovarem e de se diferenciarem em novos tipos celulares. Células tronco mesenquimais (MSC) são células tronco adultas encontradas em diferentes tecidos como tecido adiposo, polpa de dente e cordão umbilical. Estas células podem se autodividir em células idênticas ou se diferenciarem em células de origem mesodermal. Estas células têm sido estudadas em novas aplicações que envolvem terapia regenerativas. Embora resultados encorajadores tenham sido demonstrados, terapias que utilizam MSC ainda encontram uma grande barreira: a dificuldade no isolamento destas células a partir de um ambiente heterogêneo. MSC são caracterizadas por populações positivas em ensaios de imunomarcação para os epítopos membranares CD29, CD73, CD90 e CD105, presentes também em outros tipos celulares. Assim, o presente trabalho tem o objetivo de identificar novos biomarcadores de MSC de origem humana, utilizando aptâmeros de DNA produzidos pela técnica SELEX (Systematic Evolution of Ligands by EXponential Enrichment) como ferramenta. Nossos resultados mostraram que MSC de diferentes origens ligam-se a aptâmeros (oligonucleotídeos de DNA ou RNA que atuam como ligantes específicos de alvos moleculares) de DNA candidatos que atuam no isolamento de MSC por meio da técnica FACS de separação celular, promovendo uma maior indução de diferenciação em células específicas (condrócitos, osteócitos e adipócitos) comparada com a população total de MSC. Análises de citometria de fluxo mostraram que os aptâmeros candidatos se ligam a 50% das MSC de polpa de dente e não apresentam taxa de ligação significante para fibroblastos e linfócitos de origem humana - utilizados como controles negativo. Além domais, imagens de imunofluorescência e confocal mostraram ligação na superfície da membrana de MSC e a marcação interna de monócitos a estes aptâmeros. Portanto, um aptâmero controle (CNTR APT) foi utilizado para comparar a especificidade dos aptâmeros ligados a células viáveis, mostrando a não ligação deste aptâmero a MSC. Porém, 40% da população de monócitos ligou-se ao CNTR APT. Uma normalização baseada na comparação entre as taxas de ligação entre células ligadas com aptâmeros candidatos e o aptâmero controle gerou uma taxa de especificidade entre 10-16 vezes maior para MSC contra 2,5 vezes para os monócitos. Deste modo, embora os resultados tenham mostrado uma taxa de ligação entre monócitos e aptâmeros, as MSC ligadas aos aptâmeros candidatos possuem uma maior taxa de especificidade devido a uma maior presença de antígenos que são expressos em ambas as células. Um ensaio de Pull Down seguido de espectrometria de massas foi utilizado para a identificação de biomarcadores que se ligariam aos aptâmeros candidatos, e que não seriam co-expressos por monócitos e por antígenos ligados ao aptâmero controle. Deste modo, a proteína ADAM17 foi identificada nas amostras de APT10 ligadas às MSC. Tal proteína está relacionada à inibição de uma cascata de sinalização da família de proteínas TGF, responsável pela diferenciação de MSC. Assim, MSC com maior potencial tronco deveriam expressar ADAM17 em maior quantidade. Tal proteína apresenta um sítio catalítico que demonstra interagir com o APT10, de acordo com predição Docking entre proteína e DNA. Foi identificada também, a proteína VAMP3, que pertence a um complexo proteico transmembranar responsável pelos processos de endocitose e exocitose, e que podem ter um papel importante na liberação de citocinas e outras moléculas relacionadas às respostas imune e inflamatórias. Deste modo, o APT10 identificou proteínas importantes que devem estar relacionas com a melhora de imunoterapias que utilizam MSC

18ß-glycyrrhetinic acid attenuates global cerebral ischemia/reperfusion-induced cardiac damage in C57BL/J6 mice

Abstract The aim of the present study is to investigate the cardioprotective effects of 18ß-glycyrrhetinic acid (18ß -GA) against oxidative and histological damage caused by global cerebral ischemia/ reperfusion (I/R) in C57BL/J6 mice. All male mice (n:40) were randomly divided into four groups: (1) sham-operated (Sham), (2) I/R, (3) 18ß-GA, and (4) 18ß -GA+I/R. Ischemia was not applied to the sham and 18ß-GA groups. In the I/R group, the bilateral carotid arteries were clipped for 15 min to induce ischemia, and the mice were treated with the vehicle for 10 days. In the 18ß-GA group, the mice were given 18ß-GA (100 mg/kg) for 10 days following a median incision without carotid occlusion. In the 18ß-GA+I/R group, the ischemic procedure performed to the I/R model was applied to the animals and afterwards they were intraperitoneally (i.p.) treated with 18ß-GA (100 mg/kg) for 10 days. It was found that global cerebral I/R increased TBARS levels and decreased antioxidant parameters. The 18ß-GA treatment decreased the level of TBARS and increased GSH, GPx, CAT, SOD activities. Also, the control group cardiac tissue samples were observed to have a normal histological appearance with the Hematoxylin-Eosin staining method. Histopathological damage was observed in the heart tissue samples belonging to the I/R group. The 18ß-GA treatment ameliorates oxidative and histological injury in the heart tissue after global ischemia reperfusion, and may be a beneficial alternative treatment

Stingless bee propolis, metformin, and their combination alleviate diabetic cardiomyopathy

Abstract Background and aim: Stingless bee propolis, a resinous compound processed by mandibular secretion of stingless bees, is used for maintenance of hygiene and stability of beehives. Research on stingless bee propolis shows therapeutic properties attributed to polyphenols exhibiting antioxidative, antihyperglycemic and antiischemic effect. However, the cardioprotective effect of stingless bee propolis on diabetic cardiomyopathy is unknown. Methods: Adult male Sprague Dawley rats were randomised to five groups: normal group, diabetic group, diabetic given metformin (DM+M), diabetic given propolis (DM+P) and diabetic given combination therapy (DM+M+P) and treated for four weeks. Body weight, fasting blood glucose, food and water intake were taken weekly. At the end of experiment, biomarkers of oxidative damage were measured in serum and heart tissue. Antioxidants in heart tissue were quantified. Part of left ventricle of heart was processed for histological staining including Haematoxylin and Eosin (H&E) stain for myocyte size and Masson's Trichrome (MT) stain for heart fibrosis and perivascular fibrosis. Results: Propolis alleviated features of diabetic cardiomyopathy such as myocyte hypertrophy, heart fibrosis and perivascular fibrosis associated with improvement in antioxidative status. Conclusion: This study reports beneficial effect of propolis and combination with metformin in alleviating histopathological feature of diabetic cardiomyopathy by modulating antioxidants, making propolis an emerging complementary therapy.

Cell HE staining smears and paired cell paraffin sections in detection of epithelial growth factor receptor gene of pleural fluid specimens / 中南大学学报(医学版)

OBJECTIVES@#The advanced non-small cell lung cancer (NSCLC) patients with pleural effusion have no opportunity for surgery treatment. Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) are the first-line drugs for these patients with EGFR-sensitive mutation. However, the disease progression and drug update during or after treatment of EGFR-TKIs bring more challenges and puzzles to clinical diagnosis and treatment, which inevitably requires archived pleural cell samples for EGFR re-examination or comparative study. Understanding the DNA quality of archived pleural fluid samples and effectively using archival data of pleural fluid cells are of great significance for tracing the origin of cases and basic medical research. This study aims to evaluate the consistency of EGFR mutant gene expression between the 2 methods, and to explore a reliable way for preserving cytological data and making full use of cytological archival data via cell HE staining smear and cell paraffin section.@*METHODS@#A total of 57 pleural fluid cytology cases in the Department of Pathology of China Aerospace Center Hospital from October 2014 to April 2021 were selected. Tumor cells were detected by cell HE staining smears and immunohistochemical staining for TTF-1 and Napsin A in the paired cell paraffin sections. There were more than 200 tumor cells in cell HE staining smear and the proportion of tumor cells were ≥70% in matched cell paraffin sections. Patients with 2 cell smears (one for cell data retention and the other for DNA extraction) were selected as the research subjects, and 57 pleural fluid samples were enrolled. EGFR gene mutation was detected by amplification refractory mutation system-polymerase chain reaction in 57 paired cell HE staining smears and cell paraffin sections. DNA concentration was 2 ng/μL. Cell HE smear was amplified side-by-side with DNA samples from paired cell paraffin sections. Result determination was according to the requirements of the reagent instructions. The external control cycle threshold (Ct) value of the No. 8 well of the samples to be tested was between 13 and 21, which was considered as successful and reliable samples. When the Ct value of EGFR gene mutation was <26, it was considered as positive; when the Ct value was between 26 and 29, it was critical positive; when the Ct value was equal or more than 29, it was negative. ΔCt value was the difference between mutant Ct value and externally controlled Ct value. The smaller the ΔCt value was, the better the quality of DNA of the detected sample was.@*RESULTS@#Among the 57 pleural effusion samples, 42 patients were hospitalized with pleural effusion as the first symptom, accounting for 73.7% (42/57). EGFR mutation was detected in 37 samples [64.9% (37/57)]. The mutation rate for 19del was 37.8% (14/37) while for L858R was 48.6% (18/37). Females were 56.7% (21/37) of mutation cases. The mutation consistency rate of cell HE staining smear and matched cell paraffin sections was 100%. The ΔCt values of cell HE staining smears were less than those of matched cell paraffin sections. The mutation Ct values of 37 cytological samples were statistically analyzed according to the preservation periods of the years of 2014-2015, 2016-2017, 2018-2019, and 2020-2021. There were significant differences in cell paraffin section in the years of 2014-2015 and 2016-2017 compared with the years of 2018-2019 and 2020-2021, while no significant differences were found in cell HE staining smear. Statistical analysis of externally controlled Ct values of 57 cytological samples showed that there were significant differences between cell HE staining smears and cell paraffin section in the years of 2014-2015 and 2016-2017, compared with the years of 2018-2019 and 2020-2021. The mutational Ct values of 37 paired cell blocks and smears were all <26, and the externally controlled Ct values of 57 paired cell paraffin sections and HE staining smears were all between 13 and 21.@*CONCLUSIONS@#The DNA quality of cell HE smears and matched cell paraffin section met the qualified requirements. Two methods possess show an excellent consistency in detecting EGFR mutation in NSCLC pleural fluid samples. The DNA quality of cell HE staining smear is better than that of cell paraffin sections, so cell HE staining smear can be used as important supplement of the gene test source. It should be noted that the limitation of cell HE staining smears is non-reproducibility, so multiple smears of pleural fluid are recommended to be prepared for multiple tests.

Overall distribution pattern of the intramuscular nerves of the extraocular muscles and its clinical implications / Patrón de distribución general de los nervios intramusculares de los músculos extraoculares y sus implicaciones clínicas

SUMMARY: The purpose of this study was to reveal the overall distribution pattern of the intramuscular nerves of each extraocular muscle and provide morphological guidance for the selection of the neuromuscular compartment during extraocular muscle transplantation and target localization of the botulinum toxin A injection to correct strabismus. We studied 12 Chinese head specimens that were fixed with formalin. The extraocular muscles from both sides of each head were removed, and a modified Sihler's staining technique was used to reveal the overall distribution pattern of the intramuscular nerves. We observed an intramuscular nerve-dense region formed by the intramuscular arborized branches in the semitransparent superior rectus, inferior rectus, medial rectus, lateral rectus, superior oblique, inferior oblique, and levator palpebrae superioris muscles with Sihler's staining technique. The seven extraocular muscles can each be divided into two neuromuscular compartments. The intramuscular nerve-dense regions of the superior, inferior, medial, and lateral rectus and the superior oblique, inferior oblique, and levator palpebrae superioris muscles were positioned at 33.50 % -72.72 %, 40.21 % - 66.79%, 37.92 % - 64.51 %, 31.69 % - 56.01 %, 26.35 % - 64.98 %, 40.46 % - 73.20 %, and 27.72 % - 66.07 % of the lengths of the muscle bellies, respectively, and the centers of intramuscular nerve dense regions were located at 59.50 %, 54.18 %, 51.68 %, 50.08 %, 48.38 %, 56.49 %, and 50.77 % of the length of each muscle belly, respectively. The aforementioned values are the means of the actual values. These results suggest that when the strabismus is corrected with muscle transplantation, the extraocular muscle should be transplanted based on the neuromuscular compartment, which would benefit the function of both donor and recipient muscles. The localization of these nerve dense regions is recommended as an optimal target for the injection of botulinum toxin A to treat strabismus.

RESUMEN: El objetivo de este estudio fue revelar el patrón de distribución de los nervios intramusculares de cada músculo extraocular y, proporcionar una guía morfológica para la selección del compartimento neuromuscular durante el trasplante de músculo extraocular, y la localización de la inyección de toxina botulínica A para corregir el estrabismo. Estudiamos 12 muestras de cabezas de individuos chinos fijadas en formalina. Se extrajeron los músculos extraoculares de ambos lados de cada cabeza y, se utilizó una técnica de tinción de Sihler modificada para revelar el patrón de distribución general de los nervios intramusculares. Observamos una región densa en nervios intramusculares formada por los ramos intramusculares en los músculos recto superior semitransparente, recto inferior, recto medial, recto lateral, oblicuo superior, oblicuo inferior y elevador del párpado superior con técnica de tinción de Sihler. Los siete músculos extraoculares se pueden dividir cada uno en dos compartimentos neuromusculares. Las regiones intramusculares densamente nerviosas de los músculos recto superior, inferior, medial y lateral y los músculos oblicuo superior, oblicuo inferior y elevador del párpado superior se colocaron en 33,50 % -72,72 %, 40,21 % -66,79 %, 37,92 % -64,51 % , 31,69 % -56,01 %, 26,35 % -64,98 %, 40,46 % -73,20 % y 27,72 % -66,07 % de las longitudes de los vientres musculares, respectivamente, y los centros de las regiones densamente nerviosas intramusculares se ubicaron en 59,50 %, 54,18 % , 51,68 %, 50,08 %, 48,38 %, 56,49 % y 50,77 % de la longitud de cada vientre muscular, respectivamente. Los valores antes mencionados son medios de los valores reales. Estos resultados sugieren que cuando el estrabismo se corrige con trasplante de músculo, el músculo extraocular debe trasplantarse en función del compartimento neuromuscular, lo que beneficiaría la función tanto de los músculos donantes como receptores. Se recomienda la localización de estas regiones densas en nervios, como un objetivo óptimo para la inyección de toxina botulínica A para tratar el estrabismo.

Marcello Malpighi: the nervous system under a microscope / Marcello Malpighi: o sistema nervoso sob um microscópio

ABSTRACT The longstanding study of gross anatomy experienced a considerable improvement with the advent of the microscope in the early 17th century. The representative personality of this new era certainly was Marcello Malpighi, seen as "founder of microscopic anatomy". He studied, with a rudimentary compound microscope, numerous tissues and organs of several classes of animals, as well as plants. He described, for the first time, the microscopic structure of the nervous system, identifying in the gray matter of its various levels minute elements he took as "glands". It should be reminded that the concept of "cell" (and "nerve cell") was unknown at his time. Many researchers followed, performing microscopic studies, but without better results, and Malpighi's view was maintained until the beginning of the 19th century, when new histological processing and staining techniques appeared, as well as improved microscopes.

RESUMO O estudo de longa data da anatomia macroscópica experimentou um incremento considerável com o advento do microscópio no início do século 17. A personalidade representativa dessa nova era foi, certamente, Marcello Malpighi, considerado "fundador da anatomia microscópica". Ele estudou, com um microscópio composto rudimentar, numerosos tecidos e órgãos de diversas classes de animais, assim como plantas. Descreveu, pela primeira vez, a estrutura microscópica do sistema nervoso, identificando na substância cinzenta dos vários níveis elementos de minúsculas dimensões, que denominou "glândulas". Deve-se lembrar que o conceito de "célula" (e de "célula nervosa") era desconhecido naquele tempo. Muitos pesquisadores seguiram realizando estudos microscópicos, mas sem resultados melhores, e o entendimento de Malpighi foi mantido até o início do século 19, quando apareceram técnicas histológicas novas de processamento e de coloração, assim como microscópios mais aprimorados.

Using histological staining techniques to improve visualization and interpretability of tooth cementum annulation analysis / Uso de técnicas de tinción histológica para mejorar la visualización y la interpretación del análisis de la anulación del cemento dental

SUMMARY: The aim of the present study was to evaluate the effect of different staining techniques on applicability and accuracy of tooth cementum annulation (TCA) method. Nine decalcination techniques, 8 dehydration protocols and 8 different techniques were applied in 3 teeth from the persons of a known age. Black and white, and color images of histological sections were captured. An x- ray was taken of each tooth and they were photographed. Researchers were asked to observe both black/white and color images of histological sections. Researchers were divided into two groups. The first group analyzed histological images only, and the second group had photos of teeth and X-rays. In the first group of observers (without X ray) the differences in age estimation between real and observed age were significant for 2 younger patients, but not for the oldest patient, where the observed and real values matched. Of the 6 raters, the assesments of the last 3 (that used x-ray images together with histological sections) did not differ significantly from the real values. Extensive analysis and multiple repetitions performed in the present investigation revealed that the most optimal method of decalcification for TCA method was EDTA II for a period longer than 14 days at a section thickness of 2-3mm, while the most optimal protocol for dehydration was number IV. When it comes to staining, the most optimal staining protocol used for the cemental lines visualization and counting was Crocein Scarlet/Acid Fuchsin staining and Toluidine blue staining used at semithin section. Additional use of preexperimental evaluation employing x-ray of analyzed teeth decreased the errors of age estimation.

RESUMEN: El objetivo del presente estudio fue evaluar el efecto de diferentes técnicas de tinción sobre la aplicación y precisión del método de anulación de cemento dental (TCA). Se usaron nueve técnicas de descalcinación, 8 protocolos de deshidratación y 8 técnicas diferentes en 3 dientes de personas de edad conocida. Se capturaron imágenes en blanco y negro y en color de cortes histológicos. Se tomó una radiografía de cada diente y se fotografiaron. Los investigadores observaron las imágenes en blanco y negro y en color de las secciones histológicas. Los investigadores se dividieron en dos grupos; el primer grupo analizó solo imágenes histológicas y el segundo grupo tenía fotografías de los dientes y las radiografías. En el primer grupo de observadores (sin rayos X) las diferencias en la estimación de la edad entre la edad real y la edad observada fueron significativas para 2 pacientes más jóvenes, pero no para el paciente de mayor edad, donde los valores observados y reales coincidieron. De los 6 evaluadores, las valoraciones de los 3 últimos (que utilizaron imágenes de rayos X junto con cortes histológicos) no difirieron significativamente de los valores reales. El análisis exhaustivo y las múltiples repeticiones realizadas en la presente investigación revelaron que el método de descalcificación más óptimo para el método TCA fue EDTA II durante un período superior a 14 días con un grosor de sección de 2-3 mm, mientras que el protocolo óptimo para la deshidratación fue el número IV. En lo que respecta a la tinción, el protocolo de tinción más óptimo utilizado para la visualización y el recuento de las líneas de cemento fue la tinción con croceína escarlata / fucsina ácida y la tinción con azul de toluidina utilizada en la sección semifina. El uso adicional de la evaluación pre-experimental que emplea los rayos X de los dientes analizados disminuyó los errores de estimación de la edad.

Application of iodine staining technique for tumor identification in Micro-CT of mouse model with skull base-infratemporal fossa tumor / 北京大学学报(医学版)

OBJECTIVE@#To establish an animal model with malignant tumor in the skull base-infratemporal region, and to explore the role of iodine staining technique in identifying tumor tissues with Micro-CT data.@*METHODS@#Sedation anesthesia was carried out on 12 BABL/c nude mice using inhaled isoflurane, and then WSU-HN6 cells that cultured and immortalized from human tongue squamous cell carcinoma were injected into the right infratemporal fossa via the submandibular area. The procedure was carried out under ultrasonographic guidance. The nude mice were sacrificed after 3 weeks observation. The head specimens were fixed and scanned by Micro-CT, and repeated scans were performed after staining with 3.75% compound iodine solution. Following decalcification in 20% EDTA for 2-4 weeks, the head specimens were embedded and sectioned. Hematoxylin and eosin staining and Pan-Keratin immunohistochemical staining were carried out. Bright-field microscopy and stereomicroscopy were used to visualize. The Micro-CT data were analyzed using iPlan software (Brainlab).@*RESULTS@#Non-traumatic ultrasonography was used to guide HN-6 cells injection and confirm skull-base tumor formation in all the animals. Ultrasonographic guidance reduced the risk of cervical vessel injury when transferring tumor cells into the skull base space. An obvious asymmetrical appearance was detected via ultrasonography 3 weeks after tumor cell injection. The Micro-CT analysis showed that the bone was obviously damaged on the right side of the skull base, but the soft tissue image was unrecognizable. After four days staining with compound iodine solution, the morphology of the tumor and surrounding soft tissue could be clearly identified. Hematoxylin and eosin staining showed the tumor formation of the right infratemporal fossa region accompanied by bone destruction. Human keratin immunohistochemical staining showed that the tumor tissue originated from human squamous cell carcinoma, and the polynuclear osteoclasts could be seen at the margin of the skull base bone resorption.@*CONCLUSION@#The animal model with malignant tumor in the skull base-infratemporal region could be successfully established via submandibular injection under ultrasound-guidance. Bone changes of the skull were easily observed on Micro-CT, but the tumor counter was not able to be distinguished from surrounding soft tissue. The 3.75% compound iodine staining of the head specimen could help discern the tumor and surrounding soft tissue in more details.

Application of Immunohistochemistry and Special Staining Technique in Forensic Traumatic Pathology Identification / 法医学杂志

In forensic traumatic pathology practice, immunohistochemistry and special staining technique play an important role in wound age estimation and complications of traumatic complication identification. They even play an important role in the identification of special cases, such as snakebites and insulin killings. This article reviews the application and value of immunohistochemistry and special staining techniques in forensic traumatic pathology based on the cases of forensic practice reported in literature.

Diagnóstico microbiológico en lavado broncoalveolar. Revisión de la literatura / Microbiological diagnosis in bronchoalveolar lavage. Literature review

El lavado broncoalveolar (LBA) se describió hace aproximadamente 50 años, y desde ese momento se ha venido empleando cada vez con más frecuencia, llegando a ser uno de los métodos de elección para hacer el diagnóstico microbiológico de las infecciones respiratorias bajas, pues facilita la identificación de patógenos oportunistas y no oportunistas. Su uso se incrementó paralelamente con el número de pacientes inmunocomprometidos, sobre todo a causa del SIDA y los trasplantes, situaciones en las que con frecuencia los pacientes padecen infecciones pulmonares por gérmenes oportunistas. El LBA es un procedimiento seguro que permite obtener muestras que aportan información amplia de las características celulares y microbiológicas del tracto respiratorio inferior. Para garantizar su utilidad es fundamental que la recolección, transporte, almacenamiento y procesamiento de las muestras sean óptimos. El análisis de las muestras se hace por técnicas convencionales para identificación de microorganismos, como son las tinciones y el aislamiento en medios de cultivo, y por otros métodos tales como la inmunofluorescencia, pruebas inmunológicas para la detección de antígenos y anticuerpos, y pruebas de biología molecular. En la presente revisión, se hace una actualización sobre el procedimiento de obtención, almacenamiento y transporte de las muestras de LBA, así como de las técnicas de diagnóstico microbiológico más utilizadas para identificar los principales agentes infecciosos asociados con enfermedades del tracto respiratorio inferior

Bronchoalveolar lavage (BAL) was described approximately 50 years ago and since then it has been used with increasing frequency, becoming one of the methods of choice for making the microbiological diagnosis of lower respiratory infections, as it facilitates the identification of opportunistic and non-opportunistic pathogens. Its use increased in parallel with the number of immunocompromised patients, especially due to AIDS and transplantation, situations in which patients frequently suffer from lung infections due to opportunistic germs. BAL is a safe procedure that allows obtaining samples that provide comprehensive information on the cellular and microbiological characteristics of the lower respiratory tract. Optimal collection, transport, storage and processing of samples is essential to guarantee its usefulness. Analysis of the samples is done both by conventional techniques for the identification of microorganisms, such as staining and isolation in culture media, as well as by other methods such as immunofluorescence, immunological tests for the detection of antigens and antibodies, and molecular biology assays. In this review, an update in presented on the procedure for obtaining, storing and transporting BAL samples, as well as on the most widely used microbiological diagnostic techniques to identify the main infectious agents associated with lower respiratory tract diseases

Value of the toluidine blue test as an aid to determine the biopsy site in actinic prurigo cheilitis

Actinic prurigo (AP) is a type of photodermatosis that primarily affects the Latin American mestizo population. Histologically, AP cheilitis exhibits acanthosis with spongiosis and vacuolation of the basal cell layer overlying a dense lymphocytic inflammatory infiltrate that forms well-defined lymphoid follicles. Toluidine blue is a thiazide, acidophilic, and metachromatic dye used in vivo to selectively stain the acidic components of tissues such as sulfates, carboxylates, and phosphate radicals that are incorporated into DNA and RNA. It is necessary to develop a method that allows detecting, on clinical grounds the area of the lesion in which it is more feasible to find such structures. Thus to increase the sensitivity of the biopsy, in AP cheilitis to accurately identify where the lymphoid follicles reside, based on the higher concentration of DNA in such structures and thus confirm the diagnosis. In this study, staining was positive in 85% of patients with AP cheilitis, in 14 of whom 82% lymphoid follicles were observed by histopathology. One of the pathologist's problems in establishing the diagnosis of AP is that the main histopathological characteristics are not always identified in the submitted samples because it is not easy to clinically identify the most representative site of the lesion selected for performing a biopsy. Based on our results, we propose using toluidine blue as an auxiliary method to choose a tissue sample to facilitate the diagnosis and allow clinicians to make clinical correlations between the histopathological and therapeutic findings.

A new species of Characidium (Characiformes: Crenuchidae) from the Chapada dos Veadeiros, Goiás, Brazil

A new species of Characidium is described from the tributaries of the rio Tocantinzinho, rio Tocantins basin, located in the southern portion of the Chapada dos Veadeiros, at about 1,200 meters of elevation, Goiás, Brazil. The new species can be diagnosed by an unusual combination of two apomorphic features present in distinct clades of Characidium, the presence of a scaleless isthmus in allied to with a single row of dentary teeth. Additionally, the new species has a unique color pattern of inconspicuous vertical bars disconnected from the dorsal midline, forming seven to nine square blotches along body sides, and the presence of a dark saddle-shaped mark at the dorsal-fin base. Osteologically, it can be diagnosed by having the first and second anal-fin proximal radials fused and contacting the third hemal spine, which is branched. The new species also has a peculiar, unusual variation of fin-ray counts among its congeners.(AU)

Uma nova espécie de Characidium é descrita dos riachos tributários do rio Tocantins, bacia do rio Tocantins, localizados na vertente sul da Chapada dos Veadeiros, a aproximadamente 1.200 metros de altitude, Goiás, Brasil. A nova espécie pode ser diagnosticada pela combinação não usual de dois caracteres apomórficos presentes em clados distintos de Characidium, a presença do istmo sem escama em conjunto com uma única série de dentes no dentário. Adicionalmente, a nova espécie tem um padrão de coloração único de barras verticais desconectadas na região dorsal, formando sete a nove manchas quadradas ao longo do lado do corpo, e pela presença de uma mancha em forma de sela na base da nadadeira dorsal. Osteologicamente, ela pode ser diagnosticada por possuir o primeiro e segundo radiais da nadadeira anal fusionados e em contato com o terceiro espinho hemal, que é ramificado. A espécie nova também possui uma variação peculiar e pouco usual no número de raios das nadadeiras entre os congêneres.(AU)