Complete staining of nerve fiber and myoneural junctions with acetylthiocholine and silver

We describe a combined stain for simultaneous demonstration of the preterminal axons and cholinesterase activity at myoneural junctions of mammalian muscles. This technique employs acetylthiocholine iodide as the substrate for cholinesterase activity and silver nitrate impregnation of preterminal axons. The procedure is rapid, simple and uses fresh muscles. Intramuscular nerves, preterminal axons and myoneural junctions are stained simultaneously brown or black with minimal background staining of connective tissue and muscle fibers.

Chromosomal studies on five species of the genus Leptodactylus Fitzinger, 1826 (Amphibia, Anura) using differential staining

Cytogenetic studies were carried out on five species of Leptodactylus, namely L. fuscus, L, notoaktites, L. labyrinthicus, L. ocellatus, and L. podicipinus, after standard staining, Ag-NOR and C-banding as well as BrdU incorporation for three of them. The species had 2n = 22 chromosomes and two basic karyotype patterns. Chromosome 8 was a marker bearing a secondary constriction. In all species, this secondary constriction corresponded to the Ag-NOR site. The species had centromeric C-bands in all chromosomes of the complement, but some interstitial or telomeric bands seemed to differentiate some karyotypes, either at the species or the population level. In L. ocellatus, the C-banding pattern confirmed the occurrence of a heteromorphic pericentric inversion in chromosome 8 in specimens from one of the populations. The BrdU incorporation technique showed no detectable difference in the replication patterns of the major bands in the chromosomes of L. noroaktites, L. labyrinthicus, and L. ocellatus.

Evaluation of the Possibility of Removing Staining by Repolishing Composite Resins Submitted to Artificial Aging

This in vitro research verified the possibility of eliminating staining caused by coffee and red wine in five composite resins, after being submitted to thermal cycling. Thirty-six specimens were prepared and immersed in water at 37 degrees C for 24 hours. After polishing, specimen color was measured in a spectrophotometer Cintra 10 UV (Visible Spectrometer, GBC, Braeside, VIC, Australia). All specimens were submitted to thermal cycling at temperatures of 5 and 55 degrees C with a dwell time of 1 minute, for 1,000 cycles in a 75% ethanol/water solution. After thermal cycling, the specimens were immersed in water at 37 degrees C until 7 days had elapsed from the time the specimens were prepared. All specimens were then taken to the spectrophotometer for color measurement. The specimens were divided into three groups (N = 12): distilled water (control), coffee, and red wine. For the staining process to occur on only one surface, all the sides, except one, of the surfaces were isolated with white wax. The specimens were immersed in one of the solutions at 37 degrees C for 14 days. The specimens were dried and taken to the spectrophotometer for color measurement. After this, the specimens were submitted to 20 mu m wear three times, and the color was measured after each one of the wear procedures. Calculation of the color difference was made using CIEDE2000 formula. According to the methodology used in this research, it was concluded that the staining caused by coffee and red wine was superficial and one wear of 20 mu m was sufficient to remove the discoloration.

Cytogenetic analysis of Epicauta atomaria (Meloidae) and Palembus dermestoides (Tenebrionidae) with Xyp sex determination system using standard staining, C-bands, NOR and synaptonemal complex microspreading techniques

The mitotic and meiotic chromosomes of the beetles Epicauta atomaria (Meloidae) and Palembus dermestoides (Tenebrionidae) were analysed using standard staining, C-banding and silver impregnation techniques. We determine the diploid and haploid chromosome numbers, the sex determination system and describe the chromosomal morphology, the C-banding pattern and the chromosome(s) bearing NORs (nucleolar organizer regions). Both species shown 2n = 20 chromosomes, the chromosomal meioformula 9 + Xyp, and regular chromosome segregation during anaphases I and II. The chromosomes of E. atomaria are basically metacentric or submetacentric and P. dermestoides chromosomes are submetacentric or subtelocentric. In both beetles the constitutive heterochromatin is located in the pericentromeric region in all autosomes and in the Xp chromosome; additional C-bands were observed in telomeric region of the short arm in some autosomes in P. dermestoides. The yp chromosome did not show typical C-bands in these species. As for the synaptonemal complex, the nucleolar material is associated to the 7th bivalent in E. atomaria and 3rd and 7th bivalents in P. dermestoides. Strong silver impregnated material was observed in association with Xyp in light and electron microscopy preparations in these species and this material was interpreted to be related to nucleolar material.

Differential characterization of holocentric chromosomes in triatomines (Heteroptera, Triatominae) using different staining techniques and fluorescent in situ hybridization

A comparative study of holocentric chromosomes in the triatomine species Panstrongylus megistus, Rhodnius pallescens and Triatoma infestans was carried out in order to characterize heterochromatin, rDNA active sites and nucleolar proteins. Cytological preparations of seminiferous tubules were stained by silver impregnation, C banding, fluorochromes CMA 3/DA and DAPI/DA, and fluorescent in situ hybridization (FISH) with Drosophila melanogaster 28S rDNA probe. Our results showed interesting aspects of the organization of chromatin and chromosomes in the meiotic cells of these insects. In R. pallescens, sex chromosomes (X, Y) were distinct from autosomes, when submitted to silver impregnation, C banding, CMA 3 staining, and FISH, confirming that these chromosomes bear nucleolar organizer regions (NORs). In P. megistus, two of the three sex chromosomes were CMA 3/DAPI-; at early meiotic prophase and at diakinesis, silver impregnation corresponded with FISH signals, indicating that in this species, two chromosomes (probably a sex chromosome and an autosome) bear NORs. In T. infestans, silver nitrate and FISH also stained corresponding areas on meiotic chromosomes. Our data suggest that in triatomines, in general, the number and location of NORs are species-specific. These regions may be considered important chromosome markers for comparative studies to improve the understanding of evolutionary mechanisms in these hematophagous insects. ©FUNPEC-RP.

Pattern of silver nitrate-staining during meiosis and spermiogenesis in testicular lobes of Antiteuchus tripterus (Heteroptera: Pentatomidae)

The pattern of silver nitrate (Ag)-staining differed among testicular lobes of Antiteuchus tripterus. In general, these differences are in regard to the number, size, shape, coloring intensity, and location of the stained bodies or masses, observed during meiosis and spermiogenesis. These characteristics were similar in lobes 1-3. Lobes 4-6, however, differed from each other and from lobes 1-3 as well. Because the Ag-staining method is specific for nucleolar organizing regions and nucleolar material, the observations in meiosis of lobes 1-3 suggested the presence of a single pair of nucleolar organizing region-bearing chromosomes in A. tripterus, as previously found in other Pentatomidae species. In general, the amount of Ag-stained material seen in meiosis of the testicular lobes 1-3 of A. tripterus is smaller than in the other lobes. The differences among lobes observed during spermiogenesis included a striking variation in morphology of the Ag-stained material found in the head and tail of the spermatids. Given that the key role of the nucleolar material is to participate in protein synthesis, interlobular variations seem to be related to the different functions attributed to each lobe (reproduction to lobes 1-3 and basically nutrition to lobes 4-6). To our knowledge, this is the first time that the nucleolar material was studied in each testicular lobe during spermatogenesis. The present observations encourage further studies since, in addition to being of basic biological interest, several Pentatomidae species are agricultural pests and added knowledge of their biology, mainly in reproduction, may be important for the development of control strategies. ©FUNPEC-RP.

Comparison of Two Staining Methods for Pollen Viability Studies in Sugarcane

The present work aimed to compare two staining methods for pollen viability evaluation in sugarcane. Pollen from four sugarcane genotypes were collected at three different times (6.00, 8.00 and 9.00 a.m.) and tested for viability using two staining methods (iodine and lactophenol blue). Three anthers, of each genotype were crushed in a glass slide with a drop of the respective stain (iodine 0.1 N and lactophenol blue). The percentage of pollen viability was obtained with an optic microscope (250×) and compared with the pollen germination at culture media where one raquis of each genotype was gentle shaken in a petridish. Three replicates (petri dishes) was performed for each genotype which were maintained at the temperature of 25 °C and air humidity around 95 % for 30 min. The factors (staining methods, genotypes and times) and their interactions were evaluated by the analysis of variance, F test (P < 0.01) and the means compared by the t test (P < 0.05). The lactophenol blue staining was more sensible than the iodine staining method to detect the decrease of pollen viability which occurs naturally in sugarcane. The iodine staining method was more stable and easier than lactophenol to perform the inflorescence classification at any evaluated time (6.00, 8.00 and 9.00 a. m.). Both staining methods overestimated the viability obtained by the germination at culture media when performed at 6.00 a.m. © 2012 Society for Sugar Research & Promotion.

Staining of esthetic brackets by plaque disclosing solutions

AIM: To evaluate the staining of esthetic orthodontic brackets by plaque disclosing solutions. METHODS: Two types of brackets manufactured by GAC/DENTSPLY(r) were evaluated: ceramic (n=30) and polycarbonate (n=30). The brackets were divided into 6 groups. Two control groups (n=6) were immersed in absolute ethanol: GI - ceramic brackets and GII - polycarbonate brackets. Four experimental groups (n=12) were immersed in different plaque disclosing solutions: GIII (ceramic brackets) and GIV (polycarbonate brackets) were immersed in Replak(r); GV (ceramic brackets) and GVI (polycarbonate brackets) were immersed in Replasul "S"(r). Relative quantitative analysis of the influence of plaque disclosing tablets on bracket staining was performed using reflectance spectrophotometry of stain deposition. Exploratory analysis of the data was performed using Analysis of Variance (ANOVA) in a 2x2 factorial setup (bracket x immersion) with additional treatments (controls). RESULTS: The results demonstrated that the ceramic brackets presented the highest amount of staining when Replasul "S"(r) was used (pd"0.05). However, when Replak(r) was used, no statistically significant difference was found in comparison with the control group (p>0.05). For polycarbonate brackets, staining was detected for both disclosing solutions (p>0.05). CONCLUSIONS: The disclosing solutions caused stain formation on polycarbonate brackets and, under the tested conditions, use of Replak(r) on ceramic brackets did not cause staining.

Color Stability of Resin Used for Caries Infiltration After Exposure to Different Staining Solutions

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Mineral loss and color change of enamel after bleaching and staining solutions combination

Pigments of food and beverages could affect dental bleaching efficacy. The aim of this investigation was to evaluate color change and mineral loss of tooth enamel as well as the influence of staining solutions normally used by adolescent patients undergoing home bleaching. Initial hardness and baseline color were measured on enamel blocks. Specimens were divided into five groups (n = 5): G1 (control) specimens were kept in artificial saliva throughout the experiment (3 weeks); G2 enamel was exposed to 10% carbamide peroxide for 6 h daily, and after this period, the teeth were cleaned and stored in artificial saliva until the next bleaching session; and G3, G4, and G5 received the same treatments as G2, but after bleaching, they were stored for 1 h in cola soft drink, melted chocolate, or red wine, respectively. Mineral loss was obtained by the percentage of hardness reduction, and color change was determined by the difference between the data obtained before and after treatments. Data were subjected to analysis of variance and Fisher's test (a = 0.05). G3 and G5 showed higher mineral loss (92.96 +/- 5.50 and 94.46 +/- 1.00, respectively) compared to the other groups (p = 0.05). G5 showed high-color change (9.34 +/- 2.90), whereas G1 presented lower color change (2.22 +/- 0.44) (p = 0.05). Acidic drinks cause mineral loss of the enamel, which could modify the surface and reduce staining resistance after bleaching. (C) 2013 Society of Photo-Optical Instrumentation Engineers (SPIE)

Lectinhistochemical staining of granuloma induced by bacillus Calmette- Guerin in Piaractus mesopotamicus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Standardization of metachromatic staining method of myofibrillar ATPase activity of myosin to skeletal striated muscle of mules and donkeys

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of different periods of preconditioning with saliva on adhesion of C. albicans to a denture base resin by crystal violet staining and XTT assay.

Aim: The role of saliva on Candida adhesion to biomaterials has not been clearly defined. The present study investigates whether different periods of preconditioning with saliva would influence the adhesion of Candida albicans to a denture base resin. Methods: Ninety samples of acrylic resin with smooth surfaces were made and then divided into five groups: one control without saliva, and four experimental groups conditioned in saliva for periods of 30 min, 1, 3, or 12 h. Candida adhesion was evaluated by crystal violet staining and 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-([phenylamino] carbonyl)-2H-tetrazolium-hydroxide assay. Results: The one-way analysis of variance revealed that there were no significant differences among the mean number of adherent cells or among the mean absorbance for all groups. No significant correlation was found between the two methods used for assessing Candida albicans adhesion. Conclusion: The different periods of preconditioning with saliva had no significant influence on the adhesion of Candida albicans to the denture base acrylic resin.

Effect of staining agents on color change of composites

Composite resins are materials that can present color changing when exposed to pigments. Objective: The aim of this study was to evaluate, in vitro, the color changing of composites after immersion in different substances for different periods. Material and methods: Two microhybrid composite resins: Charisma (Heraeus – Kulzer) and Opallis (FGM) were used. Red wine and acai pulp were also used as immersion medium. For this study, 32 specimens with 10 mm of diameter and 2 mm of thickness were used, divided into 4 groups: Group 1 – Opallis composite immersed in red wine solution; Group 2 – Opallis composite immersed in acai berry pulp solution; Group 3 – Charisma composite immersed in red wine solution; Group 4 – Charisma composite immersed in acai berry pulp solution. The specimens were evaluated in the following time periods: T0 – baseline, T1 – 24 hours, T2 – 48 hours, T3 – 72 hours and T4 – 96 hours. For the assessment of staining, a spectrophotometer for colorimetry was used (Color Guide 45 / 0, PCB 6807 BYK-Gardner Gerestsried GmBH, Germany), and the values obtained were transferred to a computer and recorded according to CIELAB system. Results: The data were evaluated using Kruskal- Wallis non-parametric tests with the following mean values for the immersion periods of 24, 48, 72 and 96 hours, respectively: G1 – 7.35, 7.84, 9.04,10.48; G2 – 2.92, 4.15, 4.30, 4.64; G3 – 3.14, 7.35, 8.13, 8.43, G4 – 4.49, 5.99, 6.92, 6.76. Conclusion: Red wine showed a higher tendency toward altering the composite color than acai berry pulp. In addition, no significant difference was found concerning to the behavior of the two composite resins. Concerning to the immersion time periods, significant differences were only observed among the groups in the 24 hour time period.