A Set of Screening Techniques for a Quick Overview of the Neutrophil Function.

Neutrophils are known as one of the first lines of defense in the innate immune response and can perform many particular cellular functions, such as chemotaxis, reverse migration, phagocytosis, degranulation of cytotoxic enzymes and metabolites, and release of DNA as neutrophil extracellular traps (NETs). Neutrophils not only have tightly regulated signaling themselves, but also participate in the regulation of other components of the immune system. As fresh neutrophils are terminally differentiated, short-lived, and highly variable among individuals, it is important to make the most of the collected samples. Researchers often need to perform screening assays to assess an overview of the many neutrophil functions that may be affected by specific conditions under evaluation. A set of tests following a single isolation process of normal density neutrophils was developed to address this need, seeking a balance between speed, comprehensiveness, cost, and accuracy. The results can be used to reason and guide in-depth follow-up studies. This procedure can be carried out in an average time of 4 h and includes the evaluation of cell viability, reactive oxygen species (ROS) production, real-time migration, and phagocytosis of yeast on glass slides, leaving enough cells for more detailed approaches like omics studies. Moreover, the procedure includes a way to easily observe a preliminary suggestion of NETs after fast panoptic staining observed by light microscopy, with a lack of specific markers, albeit enough to indicate if further efforts in that way would be worthwhile. The diversity of functions tested combines common points among tests, reducing the analysis time and expenses. The procedure was named NeutroFun Screen, and although having limitations, it balances the aforementioned factors. Furthermore, the aim of this work is not a definite test set, but rather a guideline that can easily be adjusted to each lab's resources and demands.

Energy and mineral metabolism of peripartum mares and foals of the Quarter Horse breed.

BACKGROUND: Several studies have demonstrated variations in peripartum blood biochemical analytes used to monitor the health status of mares and their foals of different breeds. OBJECTIVES: We aimed to characterize the physiologic changes of peripartum energy and mineral metabolism in healthy Quarter Horse mares and their neonatal foals. METHODS: Blood samples were initially collected from 17 mares on days 60 and 30 prepartum, and from mares and their foals on the day of parturition (after colostrum ingestion), and on days 15, 30, and 60 postpartum. The serum concentrations of calcium, phosphorus, magnesium, total cholesterol (T-Cho), triglyceride, beta-hydroxybutyrate (BHBA), and nonesterified fatty acids (NEFA) were measured using commercial kits. Statistical differences were analyzed using one-way ANOVA followed by Tukey's test (parametric variables) or the Kruskal-Wallis test followed by Dunn's multiple comparison test (nonparametric variables). RESULTS: Fourteen of 17 healthy Quarter Horse mares and their neonatal foals remained in the study until termination. Serum BHBA, T-Cho, and calcium concentrations in mares showed significant differences during the observation period. Serum BHBA concentrations increased from 60 days prepartum to 60 days postpartum. Postpartum serum T-Cho and calcium concentrations significantly decreased until day-30 postpartum. In the foals, NEFA, T-Cho, calcium, and magnesium concentrations significantly differed between sampling times. In the foals, serum NEFA concentrations decreased from parturition until day-60 postpartum, while serum T-Cho, calcium, and magnesium concentrations were lowest at day-30 postpartum. CONCLUSIONS: The temporal changes observed in the blood biochemical analytes of this study could be used to improve the clinical evaluation of periparturient mare and neonatal foal Quarter Horses.

Color heterogeneity and individual color changes in dentin and enamel bleached in the presence of a metallic orthodontic bracket.

OBJECTIVE: This in-vitro study evaluated the color changes in enamel and dentin bleached in the presence of an orthodontic bracket, as well as the heterogeneity of the color and contribution of each tissue to the color change (CTCC). MATERIALS AND METHODS: Enamel-dentin specimens from extracted bovine teeth were bleached before or after bracket removal or maintained without any treatment (control). The colors of the dentin and enamel were measured individually and after recombining these substrates. The changes in the color (ΔE00 ) and whitening index (ΔWID) were estimated by the color difference between the bleached specimens and those of the control. The CTCC was also calculated by recombining the tissues from different treatments. Images of specimens obtained from a stereomicroscope were used to analyze the color homogeneity within each tissue. RESULTS: The highest values of ΔE00 and ΔWID were observed in the specimens bleached in the absence of a bracket. Bleaching in the presence of a bracket resulted in enamel with a center that was whiter than the periphery. Dentin and enamel presented similar CTCC values. CONCLUSIONS: Tooth bleaching performed in the presence of a metallic bracket resulted in a reduced bleaching effect and increased color heterogeneity within both tissues. CLINICAL SIGNIFICANCE: Tooth bleaching before the orthodontic bracket removal reduces the bleaching effect in both dentin and enamel. Moreover, a heterogeneous color of enamel surface can be observed after tooth bleaching in the presence of a nickel-free metallic orthodontic bracket.