Sihler's Staining of the Thoracic Cutaneous Nerve and its Significance / Tinción de Sihler del Nervio Cutáneo Torácico y su Relevancia

SUMMARY: Our team has modified Sihler's intramuscular nerve staining method to allow for calculation of nerve density. Therefore, this study aimed to show the overall distribution pattern of the thoracic cutaneous nerves to provide a morphological basis for selecting and matching sensory reconstruction during skin flap transplantation. Twelve Chinese adult cadavers were dissected; the thoracic skin was removed, and the modified Sihler's staining method was performed. Centered around the nipple, the chest skin was divided into four regions: medial-superior, lateral-superior, lateral-inferior, and medial-inferior. The thoracic skin was not only innervated by the branches of the 1st to 7th intercostal and supraclavicular nerves, but also by a small number of nerves that directly reached the skin and passed through the pectoralis major muscle. There is a phenomenon of cross overlap between the branches of adjacent intercostal nerves. The branches of the 2nd to 7th intercostal nerves were distributed in the breast, and the branches of the lateral and anterior cutaneous branches were densely distributed around the nipple, forming a grid-like anastomosis. There was no cross-overlapping innervation between the anterior cutaneous branches on both sides. The density of nerve distribution in the four regions of the chest was in the order of the medial-superior, lateral-superior, lateral-inferior and medial-inferior region, respectively. These results may be used to map sensory regions when designing thoracic skin flaps for reconstruction surgery to obtain improved sensory recovery.

Nuestro equipo ha modificado el método de tinción nerviosa intramuscular de Sihler para permitir el cálculo de la densidad nerviosa. Por lo tanto, este estudio tuvo como objetivo mostrar el patrón de distribución general de los nervios cutáneos torácicos proporcionando una base morfológica para seleccionar y combinar la reconstrucción sensorial durante el trasplante de colgajo de piel. Se diseccionaron 12 cadáveres de individuos adultos chinos. Se eliminó la piel torácica y se realizó el método de tinción de Sihler modificado, centrada alrededor del pezón, la piel del pecho se dividió en cuatro regiones: medial- superior, lateral-superior, lateral-inferior y medial-inferior. La piel torácica no solo estaba inervada por los ramos de los nervios intercostal y supraclavicular 1º a 7º, sino también por un pequeño número de nervios que llegaban directamente a la piel y pasaban a través del músculo pectoral mayor. Existe un fenómeno de superposición cruzada entre los ramos de los nervios intercostales adyacentes. Los ramos de los nervios intercostales 2º a 7º se distribuyeron en la mama, y los ramos de los ramos cutáneos lateral y anterior se distribuyeron densamente alrededor del pezón, formando una anastomosis en forma de rejilla. No hubo inervación cruzada entre los ramos cutáneos anteriores en ambos lados. La densidad de la distribución nerviosa en las cuatro regiones del tórax estaba en el orden de región medial-superior, lateral-superior, lateral-inferior y medial-inferior, respectivamente. Estos resultados pueden ser útiles para mapear regiones sensoriales al diseñar colgajos de piel torácicos para utilizarlos en cirugía de reconstrucción y obtener así una mejor recuperación sensorial.

Two protocols for the detection of oleaginous bacteria using Oil Red O.

The selection of oleaginous bacteria, potentially applicable to biotechnological approaches, is usually carried out by different expensive and time-consuming techniques. In this study, we used Oil Red O (ORO) as an useful dye for staining of neutral lipids (triacylglycerols and wax esters) on thin-layer chromatography plates. ORO could detect minimal quantities of both compounds (detection limit, 0.0025 mg of tripalmitin or 0.005 mg of cetylpalmitate). In addition, we developed a specific, rapid, and inexpensive screening methodology to detect triacylglycerol-accumulating microorganisms grown on the agar plate. This staining methodology detected 9/13 strains with a triacylglycerol content higher than 20% by cellular dry weight. ORO did not stain polyhydroxyalkanoates-producing bacteria. The four oleaginous strains not detected by this screening methodology exhibited a mucoid morphology of their colonies. Apparently, an extracellular polymeric substance produced by these strains hampered the entry of the lipophilic dye into cells. The utilization of the developed screening methodology would allow selecting of oleaginous bacteria in a simpler and faster way than techniques usually used nowadays, based on unspecific staining protocols and spectrophotometric or chromatographic methods. Furthermore, the use of ORO as a staining reagent would easily characterize the neutral lipids accumulated by microorganisms as reserve compounds. KEY POINTS: • Oil Red O staining is specific for triacylglycerols • Oil Red O staining is useful to detect oleaginous bacteria • Fast and inexpensive staining to isolate oleaginous bacteria from the environment.

Immunohistochemistry and next-generation sequencing for NTRK fusion detection in differentiated thyroid cancer of children, adolescents and young adults / Imunohistoquímica e sequenciamento nova geração para detecção de fusão NTRK em câncer diferenciado de tireoide em crianças, adolescentes e adultos jovens

INTRODUCTION: Pan-TRK immunohistochemistry staining can assess the protein expression from NTRK gene fusions. A little is known about its utility in differentiated thyroid cancer samples from children, adolescents, and young adults patients. Objective: Investigate pan-TRK immunohistochemistry sensitivity and specificity in differentiated thyroid cancer samples from children, adolescents, and young adults patients. METHODS: Tumor samples obtained from 79 children, adolescents, and young adults patients (age <21 years) diagnosed with differentiated thyroid cancer between January, 2010 and January, 2021 were retrospectively recruited from four health centers from state of Bahia e Paraíba, Brazil. NTRK gene fusion testing of all archival FFPE tumor samples: pan-TRK immunohistochemistry staining for TRKA, TRKB and TRKC protein expression were performed and then analyzed with RNA-based next-generation sequencing assay to confiC:\Users\yngrid.narciso\Desktop\CLIENTES\SITES\BJO\2024\5 - Maio\2024-05-02\siterm immunohistochemistry pan-TRK result and elucidate fusion partner. RESULTS: Pan-TRK immunohistochemistry: 3 of 79 cases had positive pan-TRK expression: next-generation sequencing; 4 were identified with NTRK gene fusion, pan-TRK immunohistochemistry was negative in all 4 NTRK next-generation sequencing-positive cases. 25 of 79 NTRK next-generation sequencing-negative control cases had concordant negative pan-TRK immunohistochemistry results. Therefore, our rate of false positive pan-TRK immunohistochemistry results was 3/25 (12%). The overall results for pan-TRK immunohistochemistry in our cohort of next-generation sequencing-negative cases was: (i) sensitivity (0%), (ii) specificity (96%), (iii) positive predictive value (94.7%), (iv) negative predictive value (91%). CONCLUSION: Pan-TRK immunohistochemistry was not a tissue-efficient screen for NTRK fusions in differentiated thyroid cancer from children, adolescents, and young adults patients. This is the largest cohort of from children, adolescents, and young adults differentiated thyroid cancer cases stained with pan-TRK immunohistochemistry, and it is the first to detail the sensitivity and specificity of pan-TRK immunohistochemistry regarding the data obtained by targeted RNA-based next-generation sequencing panel in differentiated thyroid cancer.

INTRODUÇÃO: A coloração imuno-histoquímica Pan-TRK pode avaliar a expressão proteica de fusões de genes NTRK. Pouco se sabe sobre sua utilidade em amostras diferenciadas de câncer de tireoide de crianças, adolescentes e adultos jovens. Objetivo: Investigar a sensibilidade e especificidade da imuno-histoquímica pan-TRK em amostras diferenciadas de câncer de tireoide de pacientes crianças, adolescentes e adultos jovens. MÉTODOS: Amostras tumorais obtidas de 79 pacientes crianças, adolescentes e adultos jovens (idade <21 anos) com diagnóstico de câncer diferenciado de tireoide entre janeiro de 2010 e janeiro de 2021 foram recrutadas, retrospectivamente, em quatro centros de saúde dos estados da Bahia e Paraíba, Brasil. Teste de fusão genética NTRK de todas as amostras de tumor FFPE arquivadas: coloração imuno-histoquímica pan-TRK para expressão da proteína TRKA, TRKB e TRKC foi realizada e depois analisada com ensaio de sequenciamento de próxima geração baseado em RNA, para confirmar o resultado imuno-histoquímico pan-TRK e elucidar o parceiro de fusão. RESULTADOS: Imunohistoquímica pan-TRK: 3 de 79 casos tiveram expressão pan-TRK positiva: sequenciamento de próxima geração; 4 foram identificados com fusão do gene NTRK, a imuno-histoquímica pan-TRK foi negativa em todos os 4 casos positivos para sequenciamento de próxima geração de NTRK. 25 dos 79 casos de controle negativo para sequenciamento de próxima geração de NTRK tiveram resultados de imuno-histoquímica pan-TRK negativos concordantes. Portanto, nossa taxa de resultados de imuno-histoquímica pan-TRK falsos positivos foi de 3/25 (12%). Os resultados gerais da imuno-histoquímica pan-TRK em nossa coorte de casos negativos para sequenciamento de próxima geração foram: (i) sensibilidade (0%), (ii) especificidade (96%), (iii) valor preditivo positivo (94,7%), (iv) valor preditivo negativo (91%). CONCLUSÃO: A imuno-histoquímica pan-TRK não foi uma triagem tecidualmente eficiente para fusões de NTRK em pacientes com câncer diferenciado de tireoide em crianças, adolescentes e adultos jovens. Esta é a maior coorte de casos de câncer diferenciado de tireoide de crianças, adolescentes e adultos jovens corados com imunohistoquímica pan-TRK, e é a primeira a detalhar a sensibilidade e especificidade da imunohistoquímica pan-TRK em relação aos dados obtidos por RNA direcionado baseado em um painel de sequenciamento de próxima geração no câncer diferenciado de tireoide.

Modified Goldner Trichrome for Non-decalcified Mineralized Tissue Plastinated and Embedded in Resin / Tricrómico de Goldner Modificado para Tejido Mineralizado no Descalcificado Plastinado e Incluido en resina

SUMMARY: Over time, Goldner's trichrome staining has been essential in paraffin soft tissue research. However, its classic application involves prior decalcification, generating disadvantages in the integrity of the samples and the interpretation of results. This study seeks to overcome the limitations associated with decalcification when applying Goldner's trichrome stain with plastic resins. It focuses on detailed visualization of non-decalcified bone and dental samples in animal models. Samples of jaw and tooth from a dog (Canis familiaris) were used, as well as tibia from a rabbit (Oryctolagus cuniculus) with a titanium dental implant and bone graft substitute. Adjustments were made to the original protocol, including a surface treatment prior to staining. Plastination and inclusion in specific plastic resins were part of the process. The microplastinated and stained samples showed optimal quality for optical microscopy. Those from dogs allowed detailed observation of the tooth-periodontal tissue relationship, while those from rabbits revealed a clear differentiation between mineralized and osteoid bone tissue. The staining made it easy to examine the precise interface between soft tissues, bone graft, and implant. The successful adaptation of Goldner's trichrome stain to specimens in plastic resins represents a significant advance in histological investigation of hard tissues. This methodology stands out as an effective tool to evaluate implants and biomaterials in animal models, providing detailed visualization without compromising the integrity of the samples. The combination of histochemistry and plastic resins offers a valuable alternative for microanatomical studies, opening new possibilities in hard tissue research and evaluation of bone structures.

A lo largo del tiempo, la tinción tricrómica de Goldner ha sido esencial en la investigación de tejidos blandos en parafina. Sin embargo, su aplicación clásica conlleva la descalcificación previa, generando desventajas en la integridad de las muestras y la interpretación de resultados. Este estudio busca superar las limitaciones asociadas con la descalcificación al aplicar la tinción tricrómica de Goldner con resinas plásticas. Se enfoca en visualizar detalladamente muestras óseas y dentales no descalcificadas en modelos animales. Se emplearon muestras de mandíbula y diente de perro (Canis familiaris), así como tibia de conejo (Oryctolagus cuniculus) con implante dental de titanio y substituto de injerto óseo. Se realizaron ajustes al protocolo original, incluyendo un tratamiento superficial previo a la tinción. La plastinación y la inclusión en resinas plásticas específicas fueron parte del proceso. Las muestras microplastinadas y teñidas mostraron una calidad óptima para microscopía óptica. Las de perro permitieron la observación detallada de la relación diente-tejido periodontal, mientras que las de conejo revelaron una clara diferenciación entre tejido óseo mineralizado y osteoide. La tinción facilitó examinar la interface precisa entre tejidos blandos, injerto óseo e implante. La adaptación exitosa de la tinción tricrómica de Goldner a muestras en resinas plásticas representa un avance significativo en la investigación histológica de tejidos duros. Esta metodología destaca como una herramienta eficaz para evaluar implantes y biomateriales en modelos animales, brindando una visualización detallada sin comprometer la integridad de las muestras. La combinación de histoquímica y resinas plásticas ofrece una alternativa valiosa para estudios microanatómicos, abriendo nuevas posibilidades en la investigación de tejidos duros y evaluación de estructuras óseas.

Staining blindly: an update on coloring techniques for fecal smears in parasitology: a scoping review.

Dye application for parasite highlighting in the Ova and Parasite exam is a common practice in parasitology diagnosis. Methods: A scoping review investigated how staining solutions interact with parasite structures. After screening 1334 papers, 35 met eligibility criteria. Results: Differentiating background from foreground in the fecal smear under light microscopy is the core of the research on this topic. Refractivity, unevenness of staining, size and temperature were explored to enhance staining protocols. Cryptosporidium spp. and Microsporidia were the main studied species. Conclusion: Studies on diagnostic efficacy outperform those that elucidate the physical-chemical interaction between dyes and parasites. An alternative approach involves technicians using computational tools to reduce subjectivity in fecal smear interpretation, deviating from conventional methods.

What is this article about? Coloring parasites during fecal exams has been widely used to find parasites in human feces. We searched for articles that could help us to answer the question: 'How do dyes give color to parasites?'. Then, we filtered the information from a total of 1334 articles to 35.What were the results? Cryptosporidium spp. and Microsporidia are microbes that can be seen only through a microscope. Researchers were interested in these two species in the last 40 years. Differentiating parasites from dirt on a glass slide is the main problem researchers are trying to solve. The way the light goes through parasites under a microscope, variation of staining, size and temperature of dyes have been explored to identify what gives better results in coloring protocols.What do the results of the study mean? Little is known about the chemical interaction between dyes and parasites. On the other hand, there are many studies on how good coloring methods are and comparing protocols. An alternative to the conventional approaches in staining parasites is the use of computational tools to reduce doubt in the exam interpretation by technicians.

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.

CYTOSPOT: Intracellular Staining and Detection of Cytokines by Flow Cytometer, an Alternative to LYMESPOT.

Intracellular cytokine staining is a versatile technique used to analyze cytokine production in individual cells by flow cytometry. This methodology has the specific advantage of enabling the simultaneous assessment of multiple phenotypic, differentiation, and functional parameters pertaining to responding T cells. This methodology applied after short-term culture of cells, followed by fixation and permeabilization make this technique ideal for the assessment of T-cell immune responses induced by different challenges. Here we describe an intracellular staining method followed by flow cytometry after cell stimulation with immune-relevant antigens for Lyme disease.

Does the composite attachment of clear aligners compromise the effectiveness of dental bleaching? An in vitro study.

INTRODUCTION: This study aimed to evaluate the effectiveness of dental bleaching with hydrogen peroxide 35% on the surface below the attachments. METHODS: Twenty-four blocks of bovine incisors were equally divided into 2 groups. The control group comprises the enamel surface free of attachments, whereas the attachment group comprises the enamel surface with attachment. Initial staining of samples was performed with black tea for 7 days; then, an attachment was made in the attachment group with Z250 resin and new staining for another 7 days in all samples. After staining, in-office bleaching was performed in both groups, and the attachments from the attachment group were removed. Color reading was performed with a spectrophotometer at all stages: initial (baseline), after 7 days of staining, after 14 days of staining, after immediate bleaching, and after 24 hours of bleaching. The color difference was calculated, and data analysis was performed using the t test for intergroup analysis. RESULTS: The results showed effective bleaching in both groups; however, there was a statistically significant difference in color change between them 24 hours after bleaching. CONCLUSION: It was concluded that the presence of attachment did not impair the action of the bleaching agent on the tooth surface.

48-month clinical evaluation of a copper-containing universal adhesive in non-carious cervical lesions: A double-blind randomised clinical trial.

OBJECTIVES: This study aimed to evaluate the effect of copper nanoparticles (CuNp) on the clinical performance of a universal adhesive system used as an etch-and-rinse or self-etch strategy. METHODS: A total of 216 class V (non-carious lesions) restorations were randomly placed in 36 subjects according to the following groups: ERcu, adhesive in etch-and-rinse with 0.1% CuNp; ERct, adhesive in etch-and-rinse without CuNp; SEcu, adhesive in self-etch with 0.1% CuNp; and Sect, adhesive in self-etch without CuNp. Restorations were evaluated at baseline and at 6, 12, 18, 36, and 48 months, using the FDI and USPHS criteria. Appropriate statistical analyses were performed (α = 0.05). RESULTS: After 48 months, 14 restorations were lost (two for ERcu, five for SEcu, and seven for SEct) and the retention rates (95% confidence interval [CI]) were 74.1% for ERcu (95% CI 61.1-83.8); 81.5% for ERct (95% CI 69.2-89.6); 64.8% (95% CI 51.5-76.2) for SEcu; and 64.8% (95% CI 51.5-76.2) for SEct, with statistical differences between SEct vs. ERct and SEcu vs. ERct (p < 0.05). No significant differences between the groups were observed when the secondary parameters were evaluated (p > 0.05). Nineteen restorations (two for ERcu, two for ERct, six for SEcu, and nine for SEct) showed minor marginal staining, and 44 restorations (7 for ERcu, 8 for ERct, 14 for SEcu, and 15 for SEct) presented minimal marginal adaptation defects. SIGNIFICANCE: This is the first long-term clinical trial to show that the addition of CuNp to a universal adhesive system does not affect clinical performance.

Evaluación del recolector de orina en la toma de muestra de líquido seminal y tinción diferencial de fluorescencia en espermatozoides / Evaluation of the urine collector in the collection of the seminal fluid sample and differential fluorescent staining method in sperm

Objetivo: Verificar que los recolectores de orina no son tóxicos para los espermatozoides. Métodos: Se evaluó la toxicidad de recipientes de material de vidrio (control) y plástico (recolector de orina) con muestras de semen de buena concentración y movilidad de espermatozoides, mediante el estudio de la movilidad espermática a intervalos de 1 hora, a temperatura ambiente, durante 4 horas, siguiendo lineamientos de la quinta edición del Manual de la Organización Mundial de la Salud para el procesamiento de muestras de semen. Se comparó la movilidad progresiva entre ambos tipos de recipientes (vidrio y plástico) y se evaluaron las características morfotintoriales de los espermatozoides con la tinción diferencial de fluorescencia modificada. Resultados: No se hallaron diferencias estadísticamente significativas (p = 0,334) entre los grupos. Los espermatozoides que emitieron una fluorescencia verde, se categorizaron como normales y presentaron una fuerte correlación (r = 1,000; p = 0,000); mientras que, las tinciones amarilla (r = -0,838, p = 0,009) y anaranjada (r = 0,940, p = 0,000), estuvieron altamente correlacionadas con anormalidades de los espermatozoides, destacándose que la fluorescencia anaranjada, tuvo una alta correlación (r = 0,940, p = 0,000) con anormalidades de la cabeza. Conclusión: El recolector de plástico no tiene efectos tóxicos sobre los espermatozoides, satisface las necesidades de recogida de la muestra de semen. Con la tinción diferencial de fluorescencia modificada, la tinción verde se asocia a espermatozoides morfológicamente normales y las amarilla y anaranjada se asocian a anormalidades de la cabeza, pieza intermedia y pieza principal de los espermatozoides(AU)

Objective: To verify that urine collectors are not toxic to sperm. Methods: The toxicity of glass (control) and plastic (urine collector) containers with semen samples of good concentration and sperm motility was evaluated by studying sperm motility at intervals of 1 hour, at room temperature, for 4 hours, following guidelines of the fifth edition of the World Health Organization Manual for the processing of semen samples. The progressive motility between both types of containers (glass and plastic) was compared and the morphotintorial characteristics of the spermatozoa were evaluated with the modified fluorescence differential stain. Results: No statistically significant differences (p = 0.334) were found between the groups. Spermatozoa that emitted a green fluorescence were categorized as normal and presented a strong correlation (r = 1,000; p = 0.000); Whereas, yellow (r = -0.838, p = 0.009) and orange (r = 0.940, p = 0.000) stains were highly correlated with sperm abnormalities, highlighting that orange fluorescence had a high correlation (r = 0.940, p = 0.000) with head abnormalities. Conclusion: The plastic collector has no toxic effects on sperm, it satisfies the needs of collection of the semen sample. With modified fluorescence differential staining, green staining is associated with morphologically normal spermatozoa and yellow and orange stains are associated with abnormalities of the head, middle piece, and main piece of sperm(AU)

CMA/DAPI Banding of Plant Chromosomes.

Chromosome banding based on base-specific fluorochromes, mainly double staining with chromomycin A3 (CMA) and 4'-6-diamidino-2-phenylindole (DAPI), has been widely used since the 1970s. This technique allows the differential staining of distinct types of heterochromatin. Afterward, the fluorochromes can be easily removed and leave the preparation ready for sequential procedures such as FISH or immunodetection. Interpretations of similar bands obtained with different techniques, however, merit certain caution. Here we present a detailed protocol for CMA/DAPI staining optimized for plant cytogenetics and call attention to the most common sources of misinterpretation of DAPI bands.

Silver Nitrate Staining of Nucleolar Organizer Regions (Ag-NORs) in Plant Chromosomes.

Silver nitrate staining to evidence the location of nucleolar organizer regions (Ag-NORs) in chromosomes is widely used as a classical method in plant cytogenetics. Here, we present the most used procedures and highlight some aspects in terms of their replicability by plant cytogeneticists. Some technical features described are materials and methods used, procedures, protocol modifications, and precautions in order to obtain positive signals. The methods to obtain Ag-NOR signals have different degrees of replicability, but do not require any sophisticated technology or equipment for their application.

Effect of Simulated Annual At-home Bleaching on Susceptibility to Staining, Translucency, and Whiteness Variations of Computer-aided Design and Computer-aided Manufacturing Monolithic Materials.

The effect of annual at-home bleaching on the color, translucency, and whiteness properties of computer-aided design and computer-aided manufacturing (CAD-CAM) monolithic materials requires investigation. The aim of the present in vitro study was to evaluate the effect of simulated annual at-home bleaching (consisting of daily applications for 10 hours a day for 14 days), for up to 3 years, on susceptibility to staining (ΔE00), translucency (ΔTP00), and whiteness (ΔWID) variations and on topography of CAD-CAM monolithic materials. Disks from the Lava Ultimate (LU), Vita Enamic (VE), IPS Empress CAD (EMP), and IPS e.max CAD (EMAX) were allocated as follows: 1) nonbleached or 2) bleached with 10% carbamide peroxide. After reading the CIE L*a*b* coordinates at baseline (R0), specimens were bleached or not and subsequently immersed for a 1-year simulated period in coffee before the next reading (R1). This process was repeated two more times, resulting in R2 and R3. The ΔE00, ΔTP00, and ΔWID between R1, R2, and R3 in relation to R0 were calculated. The surface topography was analyzed by scanning electron microscopy. In general, bleaching increased the staining susceptibility of all materials when compared to the nonbleached groups and that of the LU, VE, and EMAX over the years. Bleaching decreased the translucency of the VE in all years and over the years. When compared to the nonbleached groups, bleaching decreased the whiteness of the LU and EMAX and increased the whiteness of the EMP, while the VE was not affected. In the LU, the whiteness decreased over the years in both treatments, whereas the other materials were not affected with time. All materials showed progressive topographic changes over the years. The simulated annual at-home bleaching with 10% carbamide peroxide adversely affected the topography and the optical and/or colorimetric properties of the evaluated materials.

Perls/DAB Staining to Examine Iron Distribution in Arabidopsis Embryos.

Iron is accumulated in Arabidopsis embryos during seed maturation. Where iron localizes in seed and embryo is important information for seed research. Iron detection can be performed in an inexpensive manner using Perls staining, based on the Prussian blue complex formation. After this first step, DAB intensification can be performed in order to visualize easily where iron pools are located in isolated embryos.

Colorações de malhas de sombreamento sobre a fenologia, biometria e características físico-químicas de Physalis peruviana L. em sistema orgânico de produção / Colors of shadowing meshes on phenology, biometrics and physicochemical characteristics of Physalis peruviana L. in organic production system

Objetivou-se neste trabalho verificar a influência de distintas colorações de malhas de sombreamento sobre a fenologia, biometria e características físico-químicas de Physalis peruviana L em sistema orgânico de produção. O experimento foi realizado na área experimentalda UFFS, campus Laranjeiras do Sul ­PR, no setor de Horticultura. Como material vegetal foram utilizadas mudas de P. peruviana L. produzidas em estufa agrícola da área didática experimental. O delineamento experimental foi em blocos completamente casualizados, em esquema unifatorial, com quatro malhas de sombreamento (azul, preta, vermelha e branca) + testemunha (sem cobertura), com quatro repetições. Para a fenologia não houve influência estatística das colorações de malhas de sombreamento. Plantas com maior altura e menordiâmetro de caule foram verificadas com o uso da malha de sombreamento da coloração azul. Houve alterações sobre a firmeza de frutos, massa com cálice, diâmetro, altura e sólidos solúveis em função da cor da malha de sombreamento. Conclui-se que as malhasde sombreamento de diferentes colorações não interferem na fenologia de P. peruviana, mas alteram ascaracterísticas biométricas e físico-químicas das frutas, sendo a de coloração vermelha a que proporcionou os melhores resultados, não sendo recomendada autilização da malha de coloração azul.(AU)

The objective of this work was to verify the influence of different colors of shading meshes onthe phenology, biometry and physicochemical characteristics of Physalis peruvianain an organic production system. The experiment was carried out in the experimental area of UFFS, campus Laranjeiras do Sul -PR, in the Horticulture sector. As plant material, seedlings of Physalis peruvianaL. produced in an agricultural greenhouse in the experimental teaching area were used. The experimental design was in completely randomized blocks, in a one-factor scheme, with four shading meshes (blue, black, red andwhite) + control (no cover), with four replications. For the phenology there was no statistical influence of the shading mesh colors. Plants with greater height and smaller diameter were verified using the blue color shading mesh. There were changes in fruit firmness, mass with calyx, diameter, height and soluble solids depending on the type of shading. It is concluded in this work that shading nets of different colors do not affect the phenology of the P. peruviana, but they change the biometric and physicochemical characteristics of fruits, with red color being the ones that provide the best results, the use of the blue colored mesh is not recommended.(AU)

Investigation of Rhodnius prolixus head anatomy using microtomography.

In the last years, microtomography has proved to be a powerful technique on insects' studies, allowing a detailed view of the structures' internal with a high resolution. One of the most important advantages about the use of microtomography in these studies is the fact that the dissection is not necessary, which decreases considerably the number of samples used on the insects' research. Some insects are used constantly in studies about morphology, metamorphosis, and reproduction, because they work as a model for others, and Rhodnius prolixus is one of the most studied in this group. This insect is also one of the main insect vectors of Chagas disease that kills around 12,000 people every year in Latin America. Some studies using laboratory microtomography conventional scanners combining with the correct staining methods have proved that it could be a powerful tool in biological research, allowing the visualisation of low-density tissues. The main goal of the present work was to use staining protocols to study Rhodnius prolixus with laboratory microtomography conventional scanners. The experiments were carried out at the imaging lab in the Theoretical Biology Department, University of Vienna, using an Xradia MicroXCT and at the University of Oslo, using a Skyscan 2211.

Effect of modeling liquids on resin composite roughness and color parameters after staining and toothbrushing.

This study evaluated surface roughness, color stability, whitening index, and opacity of different types of modeling liquids for resin composite coating after exposure to staining and toothbrushing. Disc-shaped resin composite (Vittra APS, FGM) specimens were fabricated and divided into four groups (n = 10 each): control group, Composite Wetting resin (Ultradent Products), Adper Scotchbond Multipurpose adhesive (3M ESPE), and Adper Universal adhesive (3M ESPE). Surface roughness (Ra) was measured using a rugosimeter, while color stability (∆E00), whitening index (WI), and opacity (%) were measured using a spectrophotometer. Assessments were made at four time points: after polishing (baseline, T1), after immersion in red wine for 24 h (T2), and after 5,000 (T3) and 10,000 (T4) cycles of toothbrushing. Scanning electron microscopy images were captured to analyze the scratches created. The data were statistically analyzed by two-way repeated-measures analysis of variance and Tukey's honestly significant difference tests (α = .05). Modeling with the Wetting resin resulted in higher surface roughness (p < 0.05) and low color stability, which were attributable to porosities. Higher color change values were observed in the control group after staining. Both adhesives showed the lowest mean ΔE00 values (p < 0.005). WI decreased after staining, except with the use of the Universal adhesive (p < 0.005). The lowest opacity values were observed at baseline for all groups (p < 0.005). The Universal and Scotchbond adhesives had lower surface roughness, better color stability, higher WI, and the lowest opacity values after staining with red wine and toothbrushing.

Tooth color change promoted by different whitening toothpastes under alternate cycles of staining and brushing.

OBJECTIVES: To compare the effect of whitening toothpastes with different mechanisms of action on discolored teeth subjected to additional staining/or not. METHODS: One hundred twenty tooth specimens were stained for 14 days (staining broth under constant agitation and pH=7) and then allocated into the groups of toothpastes with different whitening ingredients (n=10): 1. Regular - Colgate Total 12 Clean Mint; 2. CLWI - Colgate Luminous White Instant (blue pigment); 3. CLWA - Colgate Luminous White Advanced Expert (hydrogen peroxide); 4. CLWAC - Colgate Luminous White Activated Charcoal (activated charcoal); 5. OB3D - Oral-B 3D WHITE Brilliant Fresh (abrasive); 6. TW - Teeth Whitening (charcoal powder). Specimens were submitted to two experimental models: A. Daily staining-toothbrush cycling: staining solution (5min), toothbrushing (45 strokes) and artificial saliva (3h), 2x/day, for 5 days; B. Only toothbrushing (30.000 brushing strokes). Color change was determined with a spectrophotometer (CIEDE2000 and Whiteness Index for Dentistry - WID) and statistically analyzed (α=0.05). RESULTS: For both models, ΔE00 and Δa did not differ significantly between the whitening toothpastes and the regular. All groups showed a decrease in tooth yellowness (-Δb) and an increase in WID. Group Teeth Whitening exhibited a decrease in luminosity (-ΔL). In model A, Groups did not differ significantly from the Regular in terms of ΔL (p=0.35) and Δb (p=0.74). Groups CLWI and OB3D exhibited a decrease in luminosity. Reduced redness (-Δa) occurred only in Group CLWI. In Model B, Groups OB3D (p=0.021) and CLWA (p=0.001) exhibited higher change in luminosity than in Group Teeth Whitening. All exhibited increased redness (+Δa) and lightness (+ΔL), except the Regular, CLWAC, and Teeth Whitening. Group OB3D had a significantly higher change in Δb than the Regular (p=0.021). CONCLUSIONS: Irrespective of the mechanism of action, all toothpastes reduced tooth yellowness and promoted similar overall color change. Exposure of the teeth to additional staining during the toothbrushing cycles did not influence the effect of the whitening toothpastes. CLINICAL SIGNIFICANCE: Whitening toothpastes should be tested in conditions that more closely simulate their use in a clinical scenario, in which alternate cycles of staining and brushing occur on a daily basis. However, even in such conditions, they were unable to promote a color change that differed from that of a regular toothpaste.

Immunohistochemical detection of Treponema pallidum in skin samples with clinical and histopathological correlations and Warthin-Starry staining critical analysis.

BACKGROUND: Syphilis in its different phases may be a difficult diagnosis in clinical and histopathological grounds. OBJECTIVES: The present study objectives were to evaluate the detection and tissue distribution of Treponema pallidum in skin lesions of syphilis. METHODS: A blinded diagnostic accuracy study was performed with immunohistochemistry and Warthin-Starry silver staining in skin samples from patients with syphilis and other diseases. Patients attended two tertiary hospitals between 2000 and 2019. Prevalence ratios (PR) and 95% confidence intervals (95% CI) were calculated for the association between immunohistochemistry positivity and clinical-histopathological variables. RESULTS: Thirty-eight patients with syphilis and their 40 biopsy specimens were included in the study. Thirty-six skin samples were used as non-syphilis controls. The Warthin-Starry technique was unable to accurately demonstrate bacteria in all samples. Immunohistochemistry showed spirochetes only in skin samples from patients with syphilis (24/40) with 60% sensitivity (95% CI 44.8‒75.2). Specificity was 100% and accuracy, 78.9% (95% CI 69.8‒88.1). Most cases had spirochetes in both dermis and epidermis and there was a high bacterial load. STUDY LIMITATIONS: Correlation between immunohistochemistry and clinical or histopathological characteristics was observed but was limited statistically due to the small sample size. CONCLUSIONS: Spirochetes were promptly seen in an immunohistochemistry protocol, which can contribute to the diagnosis of syphilis in skin biopsy samples. On the other hand, the Warthin-Starry technique showed to be of no practical value.

Two records of xanthism in Corallus hortulana (Serpentes: Boidae) in Bolivia with comments on the yellow, patternless morphotype / Dos registros de xantismo en Corallus hortulana (Serpentes: Boidae) en Bolivia con comentarios sobre el morfotipo amarillo sin patrón

Snakes display a wide range of skin colors and patterns, acting in ecological, behavioral, and physiological roles. Xanthism has been reported in numerous snake species, including many neotropical taxa. Corallus hortulana is a polychromatic species widely distributed throughout South America. Yellow, patternless individuals, while rare, have been recognized as within the species' polychromatic range. We report the first yellow, patternless specimens of Corallus hortulana collected in Bolivia and discuss whether these individuals fit the criteria of xanthism.(AU)

Las serpientes muestran una amplia gama de colores y patrones de piel, actuando en roles ecológicos, de comportamiento y fisiológicos. Además, se ha informado xantismo en numerosas especies de serpientes, incluidos muchos taxones neotropicales. Corallus hortulana es una especie policromática ampliamente distribuida en América del Sur. Los individuos amarillos sin patrón, aunque raros, se han reconocido dentro del rango policromático de la especie. Reportamos los primeros especímenes amarillos sin patrón de Corallus hortulana recolectados en Bolivia y discutimos si estos individuos cumplen con los criterios de xantismo.(AU)