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Abstract Background/objective Until 2010, adults underwent surgical treatment for maxillary expansion; however, with the advent of micro-implant-assisted rapid maxillary expansion (MARME), the availability of less invasive treatment options has increased. Nevertheless, individuals with severe transverse maxillary deficiency do not benefit from this therapy. This has aroused interest in creating a new device that allows the benefit of maxillary expansion for these individuals. The aim of this study was to evaluate the efficacy of three MARME models according to tension points, force distribution, and areas of concentration in the craniofacial complex when transverse forces are applied using finite element analysis. Materials and methods Digital modeling of the three MARME models was performed. Model A comprised five components: one body screw expander and four adjustable arms with rings for mini-implant insertion. These arms have an individualized height adjustment that allows MARME positioning according to the patient’s palatal anatomy, thereby preventing body screw expander collision with the lateral mucosa in severe cases of maxillary deficiency. Model B was a maxillary expander with screw rings joined to the body, and model C was similar to model B, except that model C had open rings for the insertion of the mini-implants. Through the MEF (Ansys software), the stresses, distribution, and area of concentration of the stresses were evaluated when transverse forces of 7.85 N were applied. Results The three models maintained the following pattern: model C presented weak stress peaks with limited distribution and lower concentration area, model B obtained median stress peaks with better distribution when compared to that of model C, and model A showed better stress distribution and larger concentration area. In model A, tensions were located in the lateral lamina of the pterygoid process, which is an important site for maxillary expansion. The limitation of the present study was that it did not include the periodontal tissues and muscles in the finite element method evaluation. Conclusions Model A showed the best stress distribution conditions. In cases of severe atresia, model A seems to be an excellent option.

Purpose: This two-arm parallel randomized controlled trial aimed to evaluate and compare periodontal changes due to rapid maxillary expansion (RME) using tooth-bone-borne and tooth-borne devices in growing patients via clinical examinations and cone-beam computed tomography (CBCT)., Materials and Methods: Forty-two eligible patients (aged 11-14 years; transverse maxillary deficiency, posterior crossbite) were screened and divided into two groups based on the treatment received (randomization was performed using computer-generated numeric sequences): hybrid hyrax tooth-bone-borne group (TBB) and hyrax tooth-borne group (TB). The primary outcome was the change in cortical bone thickness (by CBCT). In addition, the clinical attachment level (CAL), gingival recession, and bleeding were assessed. Both examinations were performed before and 3 months after the activation phase. Intergroup comparisons were performed using analysis of covariance (ANCOVA; P < 0.05)., Results: Twenty-one patients (12 girls and 9 boys; mean initial age, 13.3 years) were included in the TBB group and 21 (5 girls and 16 boys; mean initial age, 13.2 years) were included in the TB group. The TB group exhibited a decrease in buccal bone thickness in the first premolars and first molars at all three evaluated levels. Specifically, tooth 14 at 3 mm from the enamel-cement junction showed a significant width reduction (0.7 mm; p < 0.001), accompanied by a notable increase in palatal cortical thickness at 6 mm of enamel-cement junction (1.13 mm; p < 0.001)., Conclusions: RME resulted in buccal bone thickness reduction at the first premolar with hyrax treatment. In the molar region, both devices resulted in cortical bone alterations that were less pronounced in the TBB group., (© 2024. The Author(s), under exclusive licence to Springer Medizin Verlag GmbH, ein Teil von Springer Nature.)

The potential to regenerate a damaged body part is expressed to a different extent in animals. Echinoderms, in particular starfish, are known for their outstanding regenerating potential. Differently, humans have restricted abilities to restore organ systems being dependent on limited sources of stem cells. In particular, the potential to regenerate the central nervous system is extremely limited, explaining the lack of natural mechanisms that could overcome the development of neurodegenerative diseases and the occurrence of trauma. Therefore, understanding the molecular and cellular mechanisms of regeneration in starfish could help the development of new therapeutic approaches in humans. In this study, we tackle the problem of starfish central nervous system regeneration by examining the external and internal anatomical and behavioral traits, the dynamics of coelomocyte populations, and neuronal tissue architecture after radial nerve cord (RNC) partial ablation. We noticed that the removal of part of RNC generated several anatomic anomalies and induced behavioral modifications (injured arm could not be used anymore to lead the starfish movement). Those alterations seem to be related to defense mechanisms and protection of the wound. In particular, histology showed that tissue patterns during regeneration resemble those described in holothurians and in starfish arm tip regeneration. Flow cytometry coupled with imaging flow cytometry unveiled a new coelomocyte population during the late phase of the regeneration process. Morphotypes of these and previously characterized coelomocyte populations were described based on IFC data. Further studies of this new coelomocyte population might provide insights on their involvement in radial nerve cord regeneration., (© 2023. The Author(s).)

Objective: The aim of this randomized clinical trial was to evaluate and compare, during the first week of rapid maxillary expansion (RME), the impact caused by two types of appliances: Hyrax and Hybrid Hyrax., Methods: Forty-two patients who met the eligibility criteria (aged 11-14 years, with transverse maxillary deficiency, posterior crossbite, and presence of maxillary first premolars and first permanent molars) were selected and randomly divided into two groups: TBB GROUP (tooth-bone-borne expander), treated with Hybrid Hyrax (12 females and 9 males, mean age 13.3 ± 1.3 years), and TB GROUP (tooth-borne expander), treated with Hyrax (5 females and 16 males, mean age 13.3 ± 1.4 years). Pain and discomfort were assessed in two times: after the first day of activation (T1) and four days after, by means of the numerical rate scale and the instrument MFIQ (Mandibular Functional Impairment Questionnaire). Descriptive statistics and the Mann-Whitney test were used for comparison between groups and between sexes. A 5% significance level was adopted., Results: Both appliances had a negative impact, generating pain and discomfort, and reducing functional capacity. However, the scores obtained were of low intensity and no significant differences were observed between the groups. Considering sexes, there were statistically significant differences, with the female sex presenting higher scores for pain and functional limitation., Conclusions: Despite causing impact in pain and increase in the functional limitation, these changes were of low intensity, with no statistical difference between the groups. Females were more sensitive to the impact caused by the RME.

Recombinant adeno-associated virus (rAAV) has become one of the most promising gene delivery systems for both in vitro and in vivo applications. However, a key challenge is the lack of suitable imaging technologies to evaluate delivery, biodistribution and tropism of rAAVs and efficiently monitor disease amelioration promoted by AAV-based therapies at a whole-organ level with single-cell resolution. Therefore, we aimed to establish a new pipeline for the biodistribution analysis of natural and new variants of AAVs at a whole-brain level by tissue clearing and light-sheet fluorescence microscopy (LSFM). To test this platform, neonatal C57BL/6 mice were intravenously injected with rAAV9 encoding EGFP and, after sacrifice, brains were processed by standard immunohistochemistry and a recently released aqueous-based clearing procedure. This clearing technique required no dedicated equipment and rendered highly cleared brains, while simultaneously preserving endogenous fluorescence. Moreover, three-dimensional imaging by LSFM allowed the quantitative analysis of EGFP at a whole-brain level, as well as the reconstruction of Purkinje cells for the retrieval of valuable morphological information inaccessible by standard immunohistochemistry. In conclusion, the pipeline herein described takes the AAVs to a new level when coupled to LSFM, proving its worth as a bioimaging tool in tropism and gene therapy studies., (© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)

Objectives: To compare, using cone-beam computed tomography, the dentoskeletal changes in rapid maxillary expansion with tooth-bone-borne (Hybrid Hyrax) and tooth-borne (Hyrax) appliances., Setting and Sample Population: Forty-two patients who met the eligibility criteria (aged 11-14 years; transverse maxillary deficiency, posterior crossbite, and presence of upper first premolars and molars) were screened and allocated into two groups: HHG (treatment with Hybrid Hyrax) and HG (treatment with Hyrax)., Main Outcome Measures: The primary outcomes included nasomaxillary dimensional changes. CBCT was performed before and 3 months after the activation phase. Measurements were performed using Dolphin ® . Baseline data were compared using one-way ANOVA. For intergroup comparison, ANCOVA was used to analyze the initial age, appliance activations (mm), and mid-palatal suture maturation data as covariates. Statistical significance was set at 5%., Results: The premolar region in HHG showed increased skeletal changes than in HG, with the difference being 1.5 mm (0.5; 2.6) in the nasal cavity (P = .004), 1.4 mm (0.3; 2.5) in the nasal floor (P = .019), and 1.1 mm (0.2; 2.1) in the maxilla (P = .022). The molar region in HHG showed increased skeletal changes with the difference being 0.9 mm (0.2; 1.5) in the nasal cavity (P = .005), and 0.9 mm (0; 1.8) in the maxilla (P = .042) than in HG. Premolar inclination was higher in HG., Conclusion: Hybrid Hyrax showed more skeletal changes and fewer dental side effects, especially in the first premolar region. The amount of activation influenced the higher nasal skeletal changes in the Hybrid hyrax group., (© 2022 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

How the splicing machinery defines exons or introns as the spliced unit has remained a puzzle for 30 years. Here, we demonstrate that peripheral and central regions of the nucleus harbor genes with two distinct exon-intron GC content architectures that differ in the splicing outcome. Genes with low GC content exons, flanked by long introns with lower GC content, are localized in the periphery, and the exons are defined as the spliced unit. Alternative splicing of these genes results in exon skipping. In contrast, the nuclear center contains genes with a high GC content in the exons and short flanking introns. Most splicing of these genes occurs via intron definition, and aberrant splicing leads to intron retention. We demonstrate that the nuclear periphery and center generate different environments for the regulation of alternative splicing and that two sets of splicing factors form discrete regulatory subnetworks for the two gene architectures. Our study connects 3D genome organization and splicing, thus demonstrating that exon and intron definition modes of splicing occur in different nuclear regions., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Introduction: Temporary skeletal anchorage devices (TSADs) are used to obtain skeletal anchorage for orthodontic treatment. Their insertion in the infrazygomatic crest (IZC) allows efficient orthodontic mechanics. Different facial types have different bone configurations. We aimed to evaluate the differences in bone thicknesses in the IZC area among patients of each facial type to determine a safe zone for TSAD insertion., Methods: For this retrospective study, 86 cone-beam computed tomography (CBCT) scans were divided into 3 groups according to the facial type: group I, 24 CBCT scans of hyperdivergent patients; group II, 30 scans of neutral patients; and group III, 32 scans of hypodivergent patients. The buccal alveolar bone thickness was measured in 6 zones between the second premolar and distal root of the second molar, 5, 7, 9, and 11 mm apical to the alveolar crest., Results: The IZC areas with minimum thickness for TSAD insertion follows: group I, between first and second molars at 11 mm from the alveolar crest, mesial root of the second molar at 9 mm from the crest, and distal root of the second molar at 11 mm from the crest; groups II and III, between first and second molars at 11 mm from the crest and mesial root of the second molar at 11 mm from the crest., Conclusions: The safe zones for IZC miniscrew insertion are located 11 mm from the alveolar crest between the maxillary first and second molars and on the mesial root of the second molar for all the 3 facial types., (Copyright © 2021 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

Introduction: Orthodontic retraction of the anterior teeth is indicated when the patient has a malocclusion with protrusion of the incisors. Several mechanics are indicated to perform this retraction., Objective: The objective of this study was to compare the strains generated by four different types of retraction mechanics along the roots of the anterior teeth., Methods: A photoelastic model simulating an arch with first premolars extraction was made. Sixty retraction archwires were prepared, including fifteen for each type of mechanics evaluated: sliding, teardrop loop spring, T-loop spring and double key loop archwire. The strains were observed in two perspectives: occlusal and oblique. In the occlusal perspective, strains were compared among the six anterior teeth. From the oblique perspective, strains were compared among the thirds of the left canine root., Results: In the occlusal perspective, the teardrop loop spring mechanics presented greater strains, followed by T-loop spring, double key loop archwire and sliding mechanics. In all mechanics, strains were more concentrated in the canines than in the incisors. From the oblique perspective, the teardrop loop mechanics generated greater strains in the cervical regions of the canine, and in the apical regions, no differences were found in strains among the four types of mechanics. In the same mechanics, greater strains were present in the cervical zones., Conclusion: The teardrop loop spring retraction mechanic presented the greatest mean strain, and the sliding retraction mechanic presented the lowest mean strain on the root of anteroinferior teeth in the occlusal and oblique perspectives.

Background/objective: Until 2010, adults underwent surgical treatment for maxillary expansion; however, with the advent of micro-implant-assisted rapid maxillary expansion (MARME), the availability of less invasive treatment options has increased. Nevertheless, individuals with severe transverse maxillary deficiency do not benefit from this therapy. This has aroused interest in creating a new device that allows the benefit of maxillary expansion for these individuals. The aim of this study was to evaluate the efficacy of three MARME models according to tension points, force distribution, and areas of concentration in the craniofacial complex when transverse forces are applied using finite element analysis., Materials and Methods: Digital modeling of the three MARME models was performed. Model A comprised five components: one body screw expander and four adjustable arms with rings for mini-implant insertion. These arms have an individualized height adjustment that allows MARME positioning according to the patient's palatal anatomy, thereby preventing body screw expander collision with the lateral mucosa in severe cases of maxillary deficiency. Model B was a maxillary expander with screw rings joined to the body, and model C was similar to model B, except that model C had open rings for the insertion of the mini-implants. Through the MEF (Ansys software), the stresses, distribution, and area of concentration of the stresses were evaluated when transverse forces of 7.85 N were applied., Results: The three models maintained the following pattern: model C presented weak stress peaks with limited distribution and lower concentration area, model B obtained median stress peaks with better distribution when compared to that of model C, and model A showed better stress distribution and larger concentration area. In model A, tensions were located in the lateral lamina of the pterygoid process, which is an important site for maxillary expansion. The limitation of the present study was that it did not include the periodontal tissues and muscles in the finite element method evaluation., Conclusions: Model A showed the best stress distribution conditions. In cases of severe atresia, model A seems to be an excellent option.

Coelomocytes is the generic name for a collection of cellular morphotypes, present in many coelomate animals, and highly variable among echinoderm classes. The roles attributed to the major types of these free circulating cells present in the coelomic fluid of echinoderms include immune response, phagocytic digestion and clotting. Our main aim in this study was to characterize coelomocytes found in the coelomic fluid of Marthasterias glacialis (class Asteroidea) by using a combination of flow cytometry (FC), imaging flow cytometry (IFC) and fluorescence plus transmission electron microscopy (TEM). Two coelomocyte populations (P1 and P2) identified through flow cytometry were subsequently studied in terms of abundance, morphology, ultrastructure, cell viability and cell cycle profiles. Ultrastructurally, P2 diploid cells were present as two main morphotypes, similar to phagocytes and vertebrate thrombocytes, whereas the smaller P1 cellular population was characterized by low mitotic activity, a relatively undifferentiated cytotype and a high nucleus/cytoplasm ratio. In the present study we could not rule out possible similarities between haploid P1 cells and stem-cell types in other animals. Additionally, we report the presence of two other morphotypes in P2 that could only be detected by fluorescence microscopy, as well as a morphotype revealed via combined microscopy/FC. This integrative experimental workflow combined cells physical separation with different microscopic image capture technologies, enabling us to better tackle the characterization of the heterogeneous composition of coelomocytes populations., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Andrade, Oliveira, Guatelli, Martinez, Simões, Bispo, Ferrario, Bonasoro, Rino, Sugni, Gardner, Zilhão and Coelho.)

Introduction: The purposes of this research were to identify the buccolingual inclinations of the mandibular teeth and the mandibular symphysis remodeling that result from the orthodontic decompensation movement., Methods: The sample consisted of 30 adults with Class III dentofacial deformity, who had presurgical orthodontic treatment. Three-dimensional images were generated by cone-beam computed tomography scans at 2 different times (initial and before orthognathic surgery). Three-dimensional virtual models were obtained and superimposed using automated voxel-based registration at the mandible to evaluate B-point displacement, mandibular molar and incisor decompensation movement, and symphysis inclination and thickness. The 3-dimensional displacements of landmarks at the symphysis were quantified and visualized with color-coded maps using 3D Slicer (version 4.0; www.slicer.org) software., Results: The measurements showed high reproducibility. The patients presented mandibular incisor proclination, which was consistent with the movement of tooth decompensation caused by the presurgical orthodontic treatment. Statistically significant correlations were found between the inclination of the mandibular incisors, symphysis inclination, and B-point displacement. Regarding the thickness of the symphysis and the inclination of the incisors, no statistically significant correlation was found., Conclusions: The buccolingual orthodontic movement of the mandibular incisors with presurgical leveling is correlated with the inclination of the mandibular symphysis and repositioning of the B-point but not correlated to the thickness of the symphysis., (Copyright © 2020 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

Low doses of ionizing radiation (LDIR) activate endothelial cells inducing angiogenesis. In zebrafish, LDIR induce vessel formation in the sub-intestinal vessels during post-embryonic development and enhance the inter-ray vessel density in adult fin regeneration. Since angiogenesis is a crucial process involved in both post-embryonic development and regeneration, herein we aimed to understand whether LDIR accelerate these physiological conditions. Our data show that LDIR upregulate the gene expression of several pro-angiogenic molecules, such as flt1, kdr, angpt2a, tgfb2, fgf2 and cyr61in sorted endothelial cells from zebrafish larvae and this effect was abrogated by using a vascular endothelial growth factor receptor (VEGFR)-2 tyrosine kinase inhibitor. Irradiated zebrafish present normal indicators of developmental progress but, importantly LDIR accelerate post-embryonic development in a VEGFR-2 dependent signaling. Furthermore, our data show that LDIR do not accelerate regeneration after caudal fin amputation and the gene expression of the early stages markers of regeneration are not modulated by LDIR. Even though regeneration is considered as a recapitulation of embryonic development and LDIR induce angiogenesis in both conditions, our findings show that LDIR accelerate post-embryonic development but not regeneration. This highlights the importance of the physiological context for a specific phenotype promoted by LDIR.

At low doses, ionizing radiation activates endothelial cells and promotes angiogenesis. However, it is still unknown if other cells may contribute to this process. In this study, the effect of low-dose ionizing radiation (LDIR) in modulating the pro-angiogenic potential of adipocytes was investigated. Adipocytes are known to secrete multiple angiogenic factors and adipokines that induce angiogenesis. In this work, a confluent monolayer of 3T3-L1 pre-adipocytes was exposed to low doses (0.1 and 0.3 Gy) and to higher doses (0.5, 0.8 and 1.0 Gy), as control. Our data show that the adipocyte-conditioned media (A-CM) from mature adipocytes differentiated from low-dose irradiated pre-adipocytes presented a higher angiogenic potential, compared to mature adipocytes differentiated from sham-irradiated control preadipocytes. The vascular endothelial growth factor (VEGF)-A levels were significantly increased in A-CM from the 0.1 Gy ( P < 0.05) and 0.3 Gy ( P < 0.01) experimental conditions and a significant increase was found in response to 0.3 Gy dose of radiation for VEGF-C, angiopoietin-2 (ANG-2) and hepatocyte growth factor (HGF). Moreover, 0.3 Gy dose of radiation significantly increased the expression of matrix metalloproteinase (MMP)-2 active forms. In vitro , the A-CM from the 0.1 and 0.3 Gy doses experimental conditions significantly accelerated endothelial cell migration after an in vitro wound healing assay. Importantly, in vivo , the A-CM corresponding to the 0.3 Gy experimental condition significantly induced the growth of more blood vessels towards the inoculation area in the chick embryo chorioallantoic membrane (CAM). In conclusion, this work reveals a new mechanism by which low-dose radiation might promote angiogenesis, enhancing the angiogenic potential of A-CM.

Objective: To compare oral health-related quality of life (OHRQoL) before treatment of adults with unilateral cleft lip and palate (UCLP) and surgical Class III malocclusion, and to consider if clefts needing different orthodontic treatment protocols could influence people's self-perception., Design: Cross sectional., Setting: Cleft Lip and Palate Center and Clinic of Orthognathic Surgery from a School of Dentistry., Participants: A sample of adults with repaired nonsyndromic UCLP (n = 52) which was age- and sex-matched with a noncleft Class III malocclusion sample seeking orthognathic surgery (n = 51). In turn, the cleft group was subdivided according to treatment planning into nonsurgical orthodontic and surgical orthodontic approaches., Main Outcome Measure: The whole sample was assessed using the short-form oral health impact profile (OHIP-14), with higher scores indicating a poorer OHRQoL. Statistical comparisons were performed with Mann-Whitney U and Kruskal-Wallis tests, and effect size. Bonferroni adjustment was used for post hoc tests ( P < .017)., Results: The OHIP-14 scores of the UCLP and Class III groups were significantly different ( P = .001, η 2 = 0.108), and higher in Class III. The largest commitment was in the physical disability, physical pain, and psychological disability domains. In addition, no differences were found when the UCLP treatment planning was considered., Conclusion: Surgical Class III malocclusion have a poorer OHRQoL when compared to patients with UCLP, irrespective of whether they are treated surgically or orthodontically. Therefore, the greater commitment of OHRQoL appears to be influenced by the etiology of Class III, and not by treatment plan.

Objectives: This review aimed at evaluating changes in alveolar bone thickness after completion of orthodontic treatment., Methods: Only prospective clinical studies that reported bone thickness in adult patients undergoing non-surgical orthodontic treatment were considered eligible. MEDLINE, EMBASE and LILACS databases were searched for articles published up to July 2018., Results: A total of 12 studies met the selected criteria. Most of the studies showed that orthodontic treatment produces a reduction in bone thickness of incisors, mainly at the palatal side., Conclusion: On patients undergoing different orthodontic treatment techniques, there was a significant bone thickness reduction, mainly on the palatal side., Clinical Relevance: These findings are relevant and have to be considered in diagnosis and planning of tooth movement, in order to prevent the occurrence of dehiscence and fenestration in alveolar bone.

Background: The use of skeletal anchorage devices for maxillary protraction in patients with class III malocclusion due to deficiency in the middle third of the face has been shown to be a promising approach to treatment of these patients. The aim of this study was to evaluate the treatment of class III patients with maxillary retrusion, using orthodontic mini-implants (MI) associated with intermaxillary elastics in comparison with the rapid maxillary expansion and facemask protocol (RME/FM)., Methods: In this prospective non-randomized clinical trial, the sample of 24 participants between 7 and 12 years of age (median age of 10.0 years and interquartile range = 3.0 years), at the stage prior to the pre-pubertal growth spurt, was divided in two groups. In group facemask (FM) (n = 12), the individuals received orthopedic treatment with RME/FM. In group MI (n = 12), two mini-implants were inserted in the region close to the maxillary first molar roots, and the other two in the region of the mandibular canines. Initial and final lateral teleradiographs were taken for cephalometric evaluation of all the cases. Statistical analysis included the Mann-Whitney, Wilcoxon, and Fisher's exact tests. The level of significance was 5% (α = 0.05)., Results: Improvement was verified in the facial profile and occlusion of the participants, showing advancement of the maxilla in the two groups, with significant differences (P ≤ 0.05) between T0 and T1 in the following measurements: SNA, ANB, Wits, Co-A, Co-Gn, NAP, A-Npog, overjet, and molar relationship. There was no statistically significant intergroup difference (P > 0.05) in the cephalometric measurements evaluated, but the time of treatment was significant, and was faster for group MI., Conclusions: The protocol with mini-implants may be an option for the correction of Class III due to maxillary deficiency.

Herein we report the synthesis, characterization, and photophysical and biological evaluation of the complexes Ln(DBM)3(RPhen) (Ln = Sm, R = H; Ln = Sm, Eu, Tb, R = 5-NH2) stabilized by three β-diketonate units (DBM) and a phenanthroline (RPhen) derivative, with the aim of contributing to the development of lanthanide-based compounds with potential application as anticancer agents. The UV-vis spectra of [Sm(DBM)3(Phen)], [Sm(DBM)3(NH2Phen)], [Eu(DBM)3(NH2Phen)] and [Tb(DBM)3(NH2Phen)] measured in DMSO and PBS showed a strong absorption band centered at ca. 350 nm in both solvents. In DMSO, all lanthanide compounds except [Sm(DBM)3(Phen)] show a ligand centered emission band at ca. 520 nm. In PBS only sharp emission peaks are detected. The complexes show similar cytotoxic effects in A2780 ovarian cancer cells, presenting IC50 values at 24 h in the range 16-27 μM. The measurement of the cellular uptake of the complexes in the A2780 cells by inductively coupled plasma mass spectrometry (ICP-MS) revealed preferential accumulation at the membrane and cytoskeleton, with the exception of [Sm(DBM)3(Phen)] that presented higher accumulation in the cytosol than in the cell membranes. All the evaluated lanthanide complexes showed low nuclear uptake, although not negligible. Spectroscopic studies on the interaction of the complexes with calf thymus DNA (ctDNA) revealed a moderate affinity with apparent binding constants in the 104 M-1 range. Complexes bind DNA not by intercalation but probably by electrostatic interactions. A morphological evaluation of the cells treated with the different complexes by electron microscopy (TEM/SEM) proved that all of them induce mitochondrial alterations, which seemed more pronounced for the NH2Phen complexes. In addition, the complex [Eu(DBM)3(NH2Phen)] presented lysosomal uptake that might explain its augmented cytotoxicity.

Anterior crossbite is characterized by negative horizontal overjet between the jaws, and occurs due to skeletal, functional, and/or dental imbalances. It is often found in primary and mixed dentition stages and causes severe aesthetic, functional, and psychosocial impairment. We report the facial and occlusal effects of maxillary protraction associated with rapid maxillary expansion in three young patients with primary dentition. The proposed treatment plan consisted of protraction and rapid maxillary expansion. The mechanical device chosen was the facial mask combined with a McNamara rapid palatal expander. The anterior crossbite was corrected, and significant facial and cephalometric benefits were obtained. The treatment of maxillary protraction during the deciduous dentition stage, in cases where there is good acceptance and collaboration by patients, provides excellent clinical results and constitutes a viable treatment option., (Copyright © 2018 CEO. Published by Elsevier Masson SAS. All rights reserved.)

Objectives: To compare the pharyngeal airway dimensions in adults with skeletal Class III malocclusion and well-balanced faces, to ascertain whether sex can influence such measurements and to correlate skeletal pattern with pharyngeal dimensions., Methodology: Fifty adults were recruited and assigned to two groups: skeletal Class III (n = 25; 12 males and 13 females, mean age = 26.1 years), and skeletal Class I (n = 25; 11 males and 14 females, mean age = 26.0 years), with well-balanced faces. Pharyngeal measurements were made using cone beam computed tomography and analyzed with Dolphin Imaging software 11.5., Results: Pharyngeal sagittal area, length, retroglossal area and width were statistically greater for males. Pharyngeal volume was greater for Class III patients (p = .0245), due to enlarged male dimensions. A significant but moderate correlation was observed between pharyngeal airway volume and ANB angle (r = -.4917) and effective mandibular length (Co-Gn, r = .5181)., Conclusion: There is sexual differentiation in the pharyngeal airway morphology for Class III adults. As females present similar pharynx volume compared to a normal skeletal pattern, mandibular setback surgery should be carefully planned because of the risk of airway constriction.

Introduction: The surgically assisted rapid maxillary expansion is a procedure that reduces the resistance of the sutures correcting the posterior crossbite in adults., Objective: The aim of this study was to evaluate the periodontal status of 17 adults submitted to this procedure., Methods: The clinical attachment level (CAL), gingival recession, attached gingiva and bleeding were evaluated in maxillary first premolars and molars, central and lateral incisors of right and left sides before surgery, 5 days and 6 months after. Means, standard deviation, medians, minimum and maximum values were compared among the evaluations using the Friedman and McNemar tests., Results: There was a statistically significant increase in CAL in the right central incisor, right and left premolars and right and left molars. There was a statistically significant increase in gingival recession in the right and left premolars and molars. The amount of attached gingiva significantly decreased in right premolars and right and left molars. There was increase in bleeding in most of the teeth., Conclusion: Results indicated that the surgically assisted rapid maxillary expansion might cause alterations in periodontal tissue.

Introduction: The purposes of this study were to identify buccolingual inclinations and alveolar bone thickness in patients with Class III dentofacial deformities and to compare these measurements with those from subjects with normal occlusions to verify, based on the correlation between these 2 variables, whether the natural process of bone remodeling provides uniformity of bone thickness or whether it varies around the roots due to tooth inclination., Methods: The sample consisted of 35 adults with normal occlusions and 35 adults with Class III dentofacial deformities with no previous orthodontic treatment. Buccolingual inclinations and alveolar bone thickness were measured at 3 heights from the cementoenamel junction from 3-dimensional images generated by cone-beam computed tomography., Results: The region corresponding to the maxillary canines appeared to be thinner, and the palatal area of the maxillary central incisors and the distobuccal region of the mandibular second molars appeared to be thicker. Greater tooth inclinations were observed in the maxillary incisors and mandibular canines, and smaller tooth inclinations were observed on the buccal roots of the mandibular second molars., Conclusions: In subjects with Class III deformities, more statistically significant correlations were found between inclination and thickness in the mandibular teeth, whereas in subjects with normal occlusion, few statistically significant correlations were found between these 2 variables., (Copyright © 2017 American Association of Orthodontists. Published by Elsevier Inc. All rights reserved.)

We propose a novel single-deoxynucleoside-based assay that is easy to perform and provides accurate values for the absolute length (in units of time) of each of the cell cycle stages (G1, S and G2/M). This flow-cytometric assay takes advantage of the excellent stoichiometric properties of azide-fluorochrome detection of DNA substituted with 5-ethynyl-2'-deoxyuridine (EdU). We show that by pulsing cells with EdU for incremental periods of time maximal EdU-coupled fluorescence is reached when pulsing times match the length of S phase. These pulsing times, allowing labelling for a full S phase of a fraction of cells in asynchronous populations, provide accurate values for the absolute length of S phase. We characterized additional, lower intensity signals that allowed quantification of the absolute durations of G1 and G2 phases.Importantly, using this novel assay data on the lengths of G1, S and G2/M phases are obtained in parallel. Therefore, these parameters can be estimated within a time frame that is shorter than a full cell cycle. This method, which we designate as EdU-Coupled Fluorescence Intensity (E-CFI) analysis, was successfully applied to cell types with distinctive cell cycle features and shows excellent agreement with established methodologies for analysis of cell cycle kinetics.

Aims: We have previously shown that low-dose ionizing radiation (LDIR) induces angiogenesis but there is no evidence that it induces neovascularization in the setting of peripheral arterial disease. Here, we investigated the use of LDIR as an innovative and non-invasive strategy to stimulate therapeutic neovascularization using a model of experimentally induced hindlimb ischemia (HLI)., Methods and Results: After surgical induction of unilateral HLI, both hindlimbs of female C57BL/6 mice were sham-irradiated or irradiated with four daily fractions of 0.3 Gy, in consecutive days and allowed to recover. We demonstrate that LDIR, significantly improved blood perfusion in the murine ischemic limb by stimulating neovascularization, as assessed by laser Doppler flow, capillary density, and collateral vessel formation. LDIR significantly increased the circulating levels of VEGF, PlGF, and G-CSF, as well as the number of circulating endothelial progenitor cells (EPCs) mediating their incorporation to ischemic muscles. These effects were dependent upon LDIR exposition on the ischemic niche (thigh and shank regions). In irradiated ischemic muscles, these effects were independent of the recruitment of monocytes and macrophages. Importantly, LDIR induced a durable and simultaneous up-regulation of a repertoire of pro-angiogenic factors and their receptors in endothelial cells (ECs), as evident in ECs isolated from the irradiated gastrocnemius muscles by laser capture microdissection. This specific mechanism was mediated via vascular endothelial growth factor (VEGF) receptor signaling, since VEGF receptor inhibition abrogated the LDIR-mediated gene up-regulation and impeded the increase in capillary density. Finally, the vasculature in an irradiated non-ischemic bed was not affected and after 52 week of LDIR exposure no differences in the incidence of morbidity and mortality were seen., Conclusions: These findings disclose an innovative, non-invasive strategy to induce therapeutic neovascularization in a mouse model of HLI, emerging as a novel approach in the treatment of critical limb ischemia patients., (Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2017. For permissions please email: journals.permissions@oup.com.)

Spliceostatin A, meayamycin, and pladienolide B are small molecules that target the SF3b subunit of the spliceosomal U2 small nuclear ribonucleoprotein (snRNP). These compounds are attracting much attention as tools to manipulate splicing and for use as potential anti-cancer drugs. We investigated the effects of these inhibitors on mRNA transport and stability in human cells. Upon splicing inhibition, unspliced pre-mRNAs accumulated in the nucleus, particularly within enlarged nuclear speckles. However, a small fraction of the pre-mRNA molecules were exported to the cytoplasm. We identified the export adaptor ALYREF as being associated with intron-containing transcripts and show its requirement for the nucleo-cytoplasmic transport of unspliced pre-mRNA. In contrast, the exon junction complex (EJC) core protein eIF4AIII failed to form a stable complex with intron-containing transcripts. Despite the absence of EJC, unspliced transcripts in the cytoplasm were degraded by nonsense-mediated decay (NMD), suggesting that unspliced transcripts are degraded by an EJC-independent NMD pathway. Collectively, our results indicate that although blocking the function of SF3b elicits a massive accumulation of unspliced pre-mRNAs in the nucleus, intron-containing transcripts can still bind the ALYREF export factor and be transported to the cytoplasm, where they trigger an alternative NMD pathway., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2017. Published by The Company of Biologists Ltd.)

Single mRNA molecules can be imaged in living cells by a method that consists in genetically inserting binding sites for a bacteriophage protein in the gene of interest. The resulting reporter transgene is then integrated in the genome of cells that express the phage protein fused to a fluorescent tag. Upon transcription, binding of the fluorescent protein to its target sequence makes the RNA visible. With this approach it is possible to track, in real time, the life cycle of a precursor mRNA at the site of transcription in the nucleus and transport of mature mRNA to the cytoplasm. In order to measure the fluorescence associated with individual RNA molecules over time, we developed a semi-automated quantitative image analysis tool termed STaQTool. We describe in detail the implementation and application of the STaQTool software package, which is a generic tool able to process large 4D datasets allowing quantitative studies of different steps in gene expression.

Abnormal accumulation of aggregated α-synuclein (aSyn) is a hallmark of sporadic and familial Parkinson's disease (PD) and related synucleinopathies. Recent studies suggest a neuroprotective role of adenosine A2A receptor (A2AR) antagonists in PD. Nevertheless, the precise molecular mechanisms underlying this neuroprotection remain unclear. We assessed the impact of A2AR blockade or genetic deletion (A2AR KO) on synaptic plasticity and neuronal cell death induced by aSyn oligomers. We found that impairment of LTP associated with aSyn exposure was rescued in A2AR KO mice or upon A2AR blockade, through an NMDA receptor-dependent mechanism. The mechanisms underlying these effects were evaluated in SH-SY5Y cells overexpressing aSyn and rat primary neuronal cultures exposed to aSyn. Cell death in both conditions was prevented by selective A2AR antagonists. Interestingly, blockade of these receptors did not interfere with aSyn oligomerization but, instead, reduced the percentage of cells displaying aSyn inclusions. Altogether, our data raise the possibility that the well-documented effects of A2AR antagonists involve the control of the latter stages of aSyn aggregation, thereby preventing the associated neurotoxicity. These findings suggest that A2AR represent an important target for the development of effective drugs for the treatment of PD and related synucleinopathies., (© The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.)

Introduction:: Craniofacial pattern diagnosis is vital in Orthodontics, as it influences decision-making regarding treatment options and prognosis. Capelozza Filho proposed a subjective method for facial classification comprising five patterns: I, II, III, Long Face and Short Face., Objective:: To investigate the accuracy of a subjective classification method of facial patterns applied to adults., Methods:: A sample consisting of 52 adults was used for this study. Frontal and lateral view photographs were taken with subjects at rest position, including frontal smile. Lateral cephalometric radiographs were organized in a PowerPoint® presentation and submitted to 20 raters. Method performance was assessed by examining reproducibility with Kappa test and calculating accuracy, sensitivity and positive predictive values, for which 70% was set as critical value. The gold standard of the classification was personally set by the author of the method., Results:: Reproducibility was considered moderate (Kappa = 0.501); while accuracy, sensitivity and positive predictive values yielded similar results, but below 70%., Conclusions:: The subjective method of facial classification employed in the present study still needs to have its morphological criteria improved in order to be used to discriminate the five facial patterns.

The vast majority of human protein-coding genes contain up to 90% of non-coding sequence in the form of introns that must be removed from the primary transcripts or pre-mRNAs. Diverse forms of mRNAs encoded from a single gene are created by the differential use of splice sites and alternative splicing is rapidly evolving. Although the kinetic properties of splicing are thought to be critical for proofreading and regulatory mechanisms, tools for making direct experimental measurements of splicing rates are still limited. We recently developed a strategy that permits real-time imaging of fluorescent-labelled introns in single pre-mRNA molecules. Here we describe the software tool that we created for automatic tracking and quantification of intronic fluorescence at the site of transcription in live human cells., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Microscopy protocols that allow live-cell imaging of molecules and subcellular components tagged with fluorescent conjugates are indispensable in modern biological research. A breakthrough was recently introduced by the development of genetically encoded fluorescent tags that combined with fluorescence-based microscopic approaches of increasingly higher spatial and temporal resolution made it possible to detect single protein and nucleic acid molecules inside living cells. Here, we describe an approach to visualize single nascent pre-mRNA molecules and to measure in real time the dynamics of intron synthesis and excision.

Background: Rheumatoid arthritis (RA) is a chronic immune-mediated inflammatory disease characterized by cellular infiltration into the joints, hyperproliferation of synovial cells and bone damage. Available treatments for RA only induce remission in around 30% of the patients, have important adverse effects and its use is limited by their high cost. Therefore, compounds that can control arthritis, with an acceptable safety profile and low production costs are still an unmet need. We have shown, in vitro, that celastrol inhibits both IL-1β and TNF, which play an important role in RA, and, in vivo, that celastrol has significant anti-inflammatory properties. Our main goal in this work was to test the effect of celastrol in the number of sublining CD68 macrophages (a biomarker of therapeutic response for novel RA treatments) and on the overall synovial tissue cellularity and joint structure in the adjuvant-induced rat model of arthritis (AIA)., Methods: Celastrol was administered to AIA rats both in the early (4 days after disease induction) and late (11 days after disease induction) phases of arthritis development. The inflammatory score, ankle perimeter and body weight were evaluated during treatment period. Rats were sacrificed after 22 days of disease progression and blood, internal organs and paw samples were collected for toxicological blood parameters and serum proinflammatory cytokine quantification, as well as histopathological and immunohistochemical evaluation, respectively., Results: Here we report that celastrol significantly decreases the number of sublining CD68 macrophages and the overall synovial inflammatory cellularity, and halted joint destruction without side effects., Conclusions: Our results validate celastrol as a promising compound for the treatment of arthritis.

Viral interactions with host nucleus have been thoroughly studied, clarifying molecular mechanisms and providing new antiviral targets. Considering that African swine fever virus (ASFV) intranuclear phase of infection is poorly understood, viral interplay with subnuclear domains and chromatin architecture were addressed. Nuclear speckles, Cajal bodies, and promyelocytic leukaemia nuclear bodies (PML-NBs) were evaluated by immunofluorescence microscopy and Western blot. Further, efficient PML protein knockdown by shRNA lentiviral transduction was used to determine PML-NBs relevance during infection. Nuclear distribution of different histone H3 methylation marks at lysine's 9, 27 and 36, heterochromatin protein 1 isoforms (HP1α, HPβ and HPγ) and several histone deacetylases (HDACs) were also evaluated to assess chromatin status of the host. Our results reveal morphological disruption of all studied subnuclear domains and severe reduction of viral progeny in PML-knockdown cells. ASFV promotes H3K9me3 and HP1β foci formation from early infection, followed by HP1α and HDAC2 nuclear enrichment, suggesting heterochromatinization of host genome. Finally, closeness between DNA damage response factors, disrupted PML-NBs, and virus-induced heterochromatic regions were identified. In sum, our results demonstrate that ASFV orchestrates spatio-temporal nuclear rearrangements, changing subnuclear domains, relocating Ataxia Telangiectasia Mutated Rad-3 related (ATR)-related factors and promoting heterochromatinization, probably controlling transcription, repressing host gene expression, and favouring viral replication.

Introduction: Some patients refer to pre-banding orthodontic separation as a painful orthodontic procedure. Low-level laser therapy (LLLT) has been reported to have local analgesic effect., Objective: The aim of this single-blind study was to investigate the perception of pain caused by orthodontic elastomeric separators with and without a single LLLT application (6J)., Methods: The sample comprised 79 individuals aged between 13 and 34 years old at orthodontic treatment onset. Elastomeric separators were placed in first maxillary molars at mesial and distal surfaces and kept in place for three days. The volunteers scored pain intensity on a visual analogue scale (VAS) after 6 and 12 hours, and after the first, second and third days. One third of patients received laser applications, whereas another third received placebo applications and the remaining ones were controls. Applications were performed in a split-mouth design. Thus, three groups (laser, placebo and control) were assessed., Results: No differences were found among groups considering pain perception in all periods observed., Conclusion: The use of a single-dose of LLLT did not cause significant reduction in orthodontic pain perception. Overall pain perception due to orthodontic separator placement varied widely and was usually mild.

Expression of genetic information in eukaryotes involves a series of interconnected processes that ultimately determine the quality and amount of proteins in the cell. Many individual steps in gene expression are kinetically coupled, but tools are lacking to determine how temporal relationships between chemical reactions contribute to the output of the final gene product. Here, we describe a strategy that permits direct measurements of intron dynamics in single pre-mRNA molecules in live cells. This approach reveals that splicing can occur much faster than previously proposed and opens new avenues for studying how kinetic mechanisms impact on RNA biogenesis., (© 2015 Elsevier Inc. All rights reserved.)