Discovery of new ERRγ agonists regulating dopaminergic neuronal phenotype in SH-SY5Y cells.

The discovery of small molecules that regulate specific neuronal phenotypes is important for the development of new therapeutic candidates for neurological diseases. Estrogen-related receptor γ (ERRγ), an orphan nuclear receptor widely expressed in the central nervous system (CNS), is closely related to the regulation of neuronal metabolism and differentiation. We previously reported that upregulation of ERRγ could enhance dopaminergic neuronal phenotypes in the neuroblastoma cell line, SH-SY5Y. In this study, we designed and synthesized a series of new ERRγ agonists using the X-ray crystal structure of the GSK4716-bound ERRγ complex and known synthetic ligands. Our new ERRγ agonists exhibited increased transcriptional activities of ERRγ. In addition, our molecular docking results supported the experimental findings for ERRγ agonistic activity of the potent analogue, 5d. Importantly, 5d not only enhanced the expression of dopaminergic neuronal-specific molecules, TH and DAT but also activated the relevant signaling events, such as the CREB-mediated signaling pathway. The results of the present study may provide useful clues for the development of novel ERRγ agonists for neurological diseases related to the dopaminergic nervous system.

ERRγ ligand HPB2 upregulates BDNF-TrkB and enhances dopaminergic neuronal phenotype.

Brain derived neurotrophic factor (BDNF) promotes maturation of dopaminergic (DAergic) neurons in the midbrain and positively regulates their maintenance and outgrowth. Therefore, understanding the mechanisms regulating the BDNF signaling pathway in DAergic neurons may help discover potential therapeutic strategies for neuropsychological disorders associated with dysregulation of DAergic neurotransmission. Because estrogen-related receptor gamma (ERRγ) is highly expressed in both the fetal nervous system and adult brains during DAergic neuronal differentiation, and it is involved in regulating the DAergic neuronal phenotype, we asked in this study whether ERRγ ligand regulates BDNF signaling and subsequent DAergic neuronal phenotype. Based on the X-ray crystal structures of the ligand binding domain of ERRγ, we designed and synthesized the ERRγ agonist, (E)-4-hydroxy-N'-(4-(phenylethynyl)benzylidene)benzohydrazide (HPB2) (Kd value, 8.35 µmol/L). HPB2 increased BDNF mRNA and protein levels, and enhanced the expression of the BDNF receptor tropomyosin receptor kinase B (TrkB) in human neuroblastoma SH-SY5Y, differentiated Lund human mesencephalic (LUHMES) cells, and primary ventral mesencephalic (VM) neurons. HPB2-induced upregulation of BDNF was attenuated by GSK5182, an antagonist of ERRγ, and siRNA-mediated ERRγ silencing. HPB2-induced activation of extracellular-signal-regulated kinase (ERK) and phosphorylation of cAMP-response element binding protein (CREB) was responsible for BDNF upregulation in SH-SY5Y cells. HPB2 enhanced the DAergic neuronal phenotype, namely upregulation of tyrosine hydroxylase (TH) and DA transporter (DAT) with neurite outgrowth, both in SH-SY5Y and primary VM neurons, which was interfered by the inhibition of BDNF-TrkB signaling, ERRγ knockdown, or blockade of ERK activation. HPB2 also upregulated BDNF and TH in the striatum and induced neurite elongation in the substantia nigra of mice brain. In conclusion, ERRγ activation regulated BDNF expression and the subsequent DAergic neuronal phenotype in neuronal cells. Our results might provide new insights into the mechanism underlying the regulation of BDNF expression, leading to novel therapeutic strategies for neuropsychological disorders associated with DAergic dysregulation.

Synthesis of 13R,20-dihydroxy-docosahexaenoic acid by site-directed mutagenesis of lipoxygenase derived from Oscillatoria nigro-viridis PCC 7112.

Lipoxygenases (LOXs) are implicated in the biosynthesis of pro- and anti-inflammatory lipid mediators involved in immune cell signaling, most of which catalyze peroxidation of polyunsaturated fatty acids by distinct regio- and stereoselectivity. Current reports suggested that conserved amino acid, Gly in R-LOXs and Ala in S-LOXs, in the catalytic domain play an important role in determining the position as well as the stereochemistry of the functional group. Recently, we have confirmed that the catalytic specificity of cyanobacterial lipoxygenase, named Osc-LOX, with alanine at 296 was 13S-type toward linoleic acid, and producing a 17S- hydroxy-docosahexaenoic acid from docosahexaenoic acid (DHA). Here, we aimed to change the catalytic property of LOX from13S-LOX to 9R-LOX by replacing Ala with Gly and to produce a lipid mediators different from the wild-type using DHA. Finally, we succeeded in generating human endogenous a 13R-hydroxy-docosahexaenoic acid and a 13R,20-dihydroxy-docosahexaenoic acid from DHA through an enzymatic reaction using the Osc-LOX-A296G. Our study could enable physiological studies and pharmaceutical research for the 13R,20-dihydroxy-docosahexaenoic acid.

Anatomical relationship between the maxillary posterior teeth and the sinus floor according to an anterior overbite.

OBJECTIVE: To evaluate the vertical relationship between the maxillary sinus floor (MSF) and the maxillary posterior teeth (MPT) according to an anterior overbite. SETTINGS AND SAMPLE POPULATION: The patients were divided into three groups according to an anterior overbite. The open bite group (OBG) included patients with an anterior overbite of ≤0 mm, the normal overbite group (NBG) included patients with an overbite of 0-3 mm, and the deep bite group (DBG) included patients with an overbite ≥3 mm. Thirty patients were randomly matched into each group based on age and sex. MATERIALS AND METHODS: The distances and vertical relationship between the MSF and the MPT was analysed using cone-beam computer tomography. The vertical relationship between the two was classified as either favourable or unfavourable for the intrusion of the posterior teeth. RESULTS: A higher ratio of patients in the OBG had MPT positioned close to the MSF than the DBG. Intrusion of the posterior teeth was unfavourable for an average of 87.2%, 77.2% and 70.2% molars in the OBG, NBG and DBG, respectively. CONCLUSIONS: The vertical distance between the MSF and the MPT is associated with an anterior overbite. The posterior teeth were in an unfavourable position for the orthodontic intrusion in the OBG as compared to the other groups. Additional caution is needed when planning orthodontic intrusion treatment for patients with an anterior open bite. But, there were some individual variations within each group.

Comparison of maxillary basal arch forms using the root apex in adult women with different skeletal patterns: A pilot study.

INTRODUCTION: This study aimed to establish maxillary basal arch forms using the root apices and to determine the differences in the basal arch forms in adult women with different sagittal skeletal patterns. METHODS: This retrospective study included 91 adult women, with either a Class I (n = 24), Class II Division 1 (n = 22), Class II Division 2 (n = 23), or Class III (n = 22) malocclusion, who underwent cone-beam computed tomography. Three-dimensional coordinates of the root apices were determined using the multiplanar reformation mode of OnDemand3D software (Cybermed Inc, Seoul, South Korea). Two-dimensional coordinates were converted from acquired 3-dimensional coordinates via projection on the palatal plane, and the Procrustes superimposition method was used to build the basal arch form. Finally, interroot width measurements were performed for basal arch form comparisons. RESULTS: There were significant differences among the 4 groups (P <0.05) with respect to the intercanine width. The intercanine width of Class II Division 1 group was significantly narrower than that of the other groups. The Class II Division 1 and Class II Division 2 groups tended to have tapered arch forms and squared arch forms, respectively. CONCLUSIONS: We established maxillary basal arch forms using the root apices. The Class II Division 1 group had a significantly narrower intercanine distance. The use of the root apex to depict the basal arch form seems reasonable.

Asymmetric Synthesis of (-)-6-Desmethyl-Fluvirucinine A1 via Conformationally-Controlled Diastereoselective Lactam-Ring Expansions.

The versatile synthesis of (-)-6-desmethyl-fluvirucinine A1 was accomplished at a 24% overall yield through a thirteen-step process from a known vinylpiperidine. The key part involved the elaboration of the distal stereocenters and a macrolactam skeleton via conformationally-induced diastereocontrol and the iterative aza-Claisen rearrangements of lactam precursors.

Conversion of Medium-Sized Lactams to α-Vinyl or α-Acetylenyl Azacycles via N,O-Acetal TMS Ethers.

α-Vinyl or α-acetylenyl azacycles were easily synthesized from 7- to 9-membered lactams and 6- to 9-membered lactams via N,O-acetal trimethylsilyl (TMS) ethers. Organocopper and organostannane reagents afforded reasonable yields for the respective N-acyliminium ion vinylation and acetylenylation intermediates generated from N,O-acetal TMS ethers in the presence of a Lewis acid.

Conformation-Enabled Total Syntheses of Ohmyungsamycins†A and B and Structural Revision of Ohmyungsamycin†B.

The first total syntheses of the bioactive cyclodepsipeptides ohmyungsamycin A and B are described. Key features of our synthesis include the concise preparation of a linear cyclization precursor that consists of N-methyl amides and non-proteinogenic amino acids, and its macrolactamization from a bent conformation. The proposed structure of ohmyungsamycin B was revised based on its synthesis. The cyclic core of the ohmyungsamycins was shown to be responsible for the excellent antituberculosis activity, and ohmyungsamycin variants with truncated chains were evaluated for their biological activity.

Preliminary Study to Determine the Reference Plane of Patients With a Unilateral Cleft Lip and Palate.

OBJECTIVE: The objective of this study was to determine the three-dimensional midsagittal reference planes for unilateral cleft lip and palate (UCLP) patients that can be easily applied in a clinical setting. DESIGN: This was a retrospective analysis. PATIENTS: There were 35 UCLP patients (25 men, 10 women; 28.1 ± 6.9 years old) in this study. METHODS: With landmark's three-dimensional coordinates obtained from cone-beam computed tomography, the symmetric midsagittal reference planes were calculated by applying the ordinary Procrustes superimposition method using the original and mirror images. Procrustes analysis was also used to find the closest landmarks to the calculated symmetric midsagittal reference plane and test its compatibility with the symmetrical midsagittal reference plane. MAIN OUTCOME MEASURE: The three nearest landmarks to the symmetric midsagittal reference plane were Opisthion, Basion, and Nasion. RESULTS: The averages of the sums of the squared Euclidean distance and squared Procrustes distance differences between the two configurations and shapes fabricated by the symmetrical and landmark-based midsagittal reference planes, respectively, were calculated as 1.836 ± 3.295 and 1.519 × 10-5 ± 2.351 × 10-5. CONCLUSION: It was confirmed that the midsagittal reference planes from these selected landmarks for UCLP patients were compatible with symmetric midsagittal reference planes from the Procrustes analysis and the asymmetric measurements.

Assessment of the root apex position of impacted maxillary canines on panoramic films.

INTRODUCTION: In this study, we aimed to analyze the root apex positions of impacted maxillary canines on panoramic radiographs according to their labiopalatal positions and to propose a new panoramic radiographic predictor of impacted maxillary canines. METHODS: Paired panoramic films and cone-beam computed tomography images of 104 patients with unilateral impacted maxillary canines were selected. The sector locations of the impacted canine root apices on the panoramic radiographs were compared with the labiopalatal positions of impacted maxillary canines on cone-beam computed tomography. Statistical correlations between panoramic and cone-beam computed tomography findings were examined with the chi-square test. RESULTS: A statistically significant association was observed between the sector locations of impacted canine root apices and the labiopalatal positions of the canines (P <0.001). The root apices of labially impacted canines were more frequently located in sector 2 (corresponding to the lateral incisor position), those of palatally impacted canines were more frequently located in sector 4 (corresponding to the first premolar), and those of midalveolus impacted canines were more frequently located in sector 3 (corresponding to the canine). CONCLUSIONS: On panoramic films, the root apices of palatally impacted canines tended toward the root apices of the maxillary first premolars, whereas those of labially impacted canines tended toward the root apices of the maxillary lateral incisors. Therefore, the sector locations of impacted canine root apices on panoramic radiographs could be used for the early prediction of maxillary canine impactions and determination of the labiopalatal positions of impacted canines.

How far is the root apex of a unilateral impacted canine from the root apices' arch form?

INTRODUCTION: The purpose of this study was to determine the arch form of the root apices of normally erupting teeth and then determine the differences in the location of the apex of impacted canines relative to normally erupting canines. In addition, we sought to determine whether the labiopalatal position of the impacted canines influences the position of the apices. METHODS: The study included 21 patients with unerupted canines that subsequently had a normal eruption, 21 patients with palatally impacted canines, 27 patients with labially impacted canines, and 17 patients with midalveolus impacted canines. Images were obtained using cone beam computed tomography, and the x, y, and z coordinates of the root apices were determined using Ondemand3D software (Cybermed Co., Seoul, Korea). Two-dimensional coordinates were converted from acquired 3-dimensional coordinates via projection on a palatal plane, and the Procrustes method was used to process the converted 2-dimensional coordinates and to draw the arch forms of the root apices. Finally, we measured the extent of root apex deviation from the arch forms of the root apices. RESULTS: Normally erupting canines showed that even though calcifications may be immature, their positions were aligned with a normal arch form. The root apices of the impacted canines were an average of 6.572 mm away from the root apices' arch form, whereas those of the contralateral nonimpacted canines were an average distance of 2.221 mm away, a statistically significant difference. The palatally impacted canines' root apices distribution tended toward the first premolar root apices. CONCLUSIONS: Incompletely calcified, unerupted teeth with a subsequent normal eruption showed a normal arch form of the root apices. The root apices of impacted canines were farther from the arch forms than were the nonimpacted canines. Also, the root apices of impacted canines in the palatal area showed distributions different from those of the other impacted canine groups.

Matrix metalloproteinase-8 plays a pivotal role in neuroinflammation by modulating TNF-α activation.

Matrix metalloproteinases (MMPs) play important roles in normal brain development and synaptic plasticity, although aberrant expression of MMPs leads to brain damage, including blood-brain barrier disruption, inflammation, demyelination, and neuronal cell death. In this article, we report that MMP-8 is upregulated in LPS-stimulated BV2 microglial cells and primary cultured microglia, and treatment of MMP-8 inhibitor (M8I) or MMP-8 short hairpin RNA suppresses proinflammatory molecules, particularly TNF-α secretion. Subsequent experiments showed that MMP-8 exhibits TNF-α-converting enzyme (TACE) activity by cleaving the prodomain of TNF-α (A(74)/Q(75), A(76)/V(77) residues) and, furthermore, that M8I inhibits TACE activity more efficiently than TAPI-0, a general TACE inhibitor. Biochemical analysis of the underlying anti-inflammatory mechanisms of M8I revealed that it inhibits MAPK phosphorylation, NF-κB/AP-1 activity, and reactive oxygen species production. Further support for the proinflammatory role of microglial MMP-8 was obtained from an in vivo animal model of neuroinflammatory disorder. MMP-8 is upregulated in septic conditions, particularly in microglia. Administration of M8I or MMP-8 short hairpin RNA significantly inhibits microglial activation and expression/secretion of TNF-α in brain tissue, serum, and cerebrospinal fluid of LPS-induced septic mice. These results demonstrate that MMP-8 critically mediates microglial activation by modulating TNF-α activity, which may explain neuroinflammation in septic mouse brain.

Phosphorylation of hepatitis C virus RNA polymerases ser29 and ser42 by protein kinase C-related kinase 2 regulates viral RNA replication.

UNLABELLED: Hepatitis C virus (HCV) nonstructural protein 5B (NS5B), an RNA-dependent RNA polymerase (RdRp), is the key enzyme for HCV RNA replication. We previously showed that HCV RdRp is phosphorylated by protein kinase C-related kinase 2 (PRK2). In the present study, we used biochemical and reverse-genetics approaches to demonstrate that HCV NS5B phosphorylation is crucial for viral RNA replication in cell culture. Two-dimensional phosphoamino acid analysis revealed that PRK2 phosphorylates NS5B exclusively at its serine residues in vitro and in vivo. Using in vitro kinase assays and mass spectrometry, we identified two phosphorylation sites, Ser29 and Ser42, in the Δ1 finger loop region that interacts with the thumb subdomain of NS5B. Colony-forming assays using drug-selectable HCV subgenomic RNA replicons revealed that preventing phosphorylation by Ala substitution at either Ser29 or Ser42 impairs HCV RNA replication. Furthermore, reverse-genetics studies using HCV infectious clones encoding phosphorylation-defective NS5B confirmed the crucial role of these PRK2 phosphorylation sites in viral RNA replication. Molecular-modeling studies predicted that the phosphorylation of NS5B stabilizes the interactions between its Δ1 loop and thumb subdomain, which are required for the formation of the closed conformation of NS5B known to be important for de novo RNA synthesis. Collectively, our results provide evidence that HCV NS5B phosphorylation has a positive regulatory role in HCV RNA replication. IMPORTANCE: While the role of RNA-dependent RNA polymerases (RdRps) in viral RNA replication is clear, little is known about their functional regulation by phosphorylation. In this study, we addressed several important questions about the function and structure of phosphorylated hepatitis C virus (HCV) nonstructural protein 5B (NS5B). Reverse-genetics studies with HCV replicons encoding phosphorylation-defective NS5B mutants and analysis of their RdRp activities revealed previously unidentified NS5B protein features related to HCV replication and NS5B phosphorylation. These attributes most likely reflect potential structural changes induced by phosphorylation in the Δ1 finger loop region of NS5B with two identified phosphate acceptor sites, Ser29 and Ser42, which may transiently affect the closed conformation of NS5B. Elucidating the effects of dynamic changes in NS5B phosphorylation status during viral replication and their impacts on RNA synthesis will improve our understanding of the molecular mechanisms of NS5B phosphorylation-mediated regulation of HCV replication.

Proximal Segment Changes After Bilateral Sagittal Split Ramus Osteotomy in Facial Asymmetry Patients.

PURPOSE: To investigate the 3-dimensional postoperative changes in the proximal segments in patients with facial asymmetry according to the anteroposterior skeletal patterns. MATERIALS AND METHODS: A total of 51 patients with facial asymmetry who had undergone Le Fort I osteotomy and sagittal split ramus osteotomy (SSRO) with rigid fixation were classified according to their anteroposterior skeletal patterns. Cone-beam computed tomography (CBCT) data were obtained before (T0) and 6 months (T1) after surgery. The position, angulation, and distance of the proximal segment were measured from the CBCT superimposition. RESULTS: Comparing the T0 and T1 CBCT data, almost no significant differences were found in the condylar head position in any of the groups (P > .05), except for the axial condylar head position on the deviated side in the skeletal Class I group (P < .05) and the sagittal condylar head position on the deviated side in the skeletal Class III group (P < .05). However, the changes in the coronoid process and ramus down varied (P < .05), and these movements were related to changes in the ramal plane. Comparing the deviated and nondeviated sides, significant differences were found only in the skeletal Class I group (P < .05). CONCLUSIONS: The most influential factors affecting differences between the deviated and nondeviated sides in patients with facial asymmetry after bilateral SSRO could be the direction of the surgical movement of the distal segment of the mandible rather than its extent.

Differences Among Deviations, Genders, and Observers in the Perception of Eye and Nose Asymmetry.

PURPOSE: This study investigated the degree and range of recognition of canting of the interpupillary line and deviation of the nose, which were regarded as "normal," "acceptable," and "needing surgical correction," according to different observer groups. MATERIALS AND METHODS: Four different groups (40 each for laypeople, dental students, general dentists, and orthodontists) rated 11 simulated asymmetric images with canted eyes and a deviated nose separately. The raters categorized each image as normal, socially "acceptable" and not requiring correction, or abnormal and would benefit from correction. Survival analysis was used to evaluate the scope of perception. RESULTS: The degree of recognition for eye-canting and nose deviation was 2.31 and 2.92, respectively. Eye-canting and nose deviation had lower degrees of recognition in the clockwise direction (2.13°) and on the left side (2.65°), respectively. Women showed a lower degree of the "upper limit of the acceptable range" for canted eyes, and men showed a lower "degree of recognition" and "upper limit of the acceptable range" for a deviated nose. Orthodontists showed a larger upper limit of the acceptable range for eye-canting (P < .001). CONCLUSIONS: The perceptions of asymmetry of canted eyes and deviated nose were affected by gender, direction of asymmetry, and observer groups.

Quantitative skeletal maturation estimation using cone-beam computed tomography-generated cervical vertebral images: a pilot study in 5- to 18-year-old Japanese children.

OBJECTIVES: The purpose of this study was to establish multivariable regression models for the estimation of skeletal maturation status in Japanese boys and girls using the cone-beam computed tomography (CBCT)-based cervical vertebral maturation (CVM) assessment method and hand-wrist radiography. MATERIALS AND METHODS: The analyzed sample consisted of hand-wrist radiographs and CBCT images from 47 boys and 57 girls. To quantitatively evaluate the correlation between the skeletal maturation status and measurement ratios, a CBCT-based CVM assessment method was applied to the second, third, and fourth cervical vertebrae. Pearson's correlation coefficient analysis and multivariable regression analysis were used to determine the ratios for each of the cervical vertebrae (p < 0.05). RESULTS: Four characteristic parameters ((OH2 + PH2)/W2, (OH2 + AH2)/W2, D2, AH3/W3), as independent variables, were used to build the multivariable regression models: for the Japanese boys, the skeletal maturation status according to the CBCT-based quantitative cervical vertebral maturation (QCVM) assessment was 5.90 + 99.11 × AH3/W3 - 14.88 × (OH2 + AH2)/W2 + 13.24 × D2; for the Japanese girls, it was 41.39 + 59.52 × AH3/W3 - 15.88 × (OH2 + PH2)/W2 + 10.93 × D2. CONCLUSIONS: The CBCT-generated CVM images proved very useful to the definition of the cervical vertebral body and the odontoid process. The newly developed CBCT-based QCVM assessment method showed a high correlation between the derived ratios from the second cervical vertebral body and odontoid process. CLINICAL RELEVANCE: There are high correlations between the skeletal maturation status and the ratios of the second cervical vertebra based on the remnant of dentocentral synchondrosis.

Structural overview of toxin-antitoxin systems in infectious bacteria: a target for developing antimicrobial agents.

The bacterial toxin-antitoxin (TA) system is a module that may play a role in cell survival under stress conditions. Generally, toxin molecules act as negative regulators in cell survival and antitoxin molecules as positive regulators. Thus, the expression levels and interactions between toxins and antitoxins should be systematically harmonized so that bacteria can escape such harmful conditions. Since TA systems are able to control the fate of bacteria, they are considered potent targets for the development of new antimicrobial agents. TA systems are widely prevalent with a variety of systems existing in bacteria: there are three types of bacterial TA systems depending on the property of the antitoxin which binds either the protein toxin or mRNA coding the toxin protein. Moreover, the multiplicity of TA genes has been observed even in species of bacteria. Therefore, knowledge on TA systems such as the individual characteristics of TA systems, integrative working mechanisms of various TA systems in bacteria, interactions between toxin molecules and cellular targets, and so on is currently limited due to their complexity. In this regard, it would be helpful to know the structural characteristics of TA modules for understanding TA systems in bacteria. Until now, 85 out of the total structures deposited in PDB have been bacterial TA system proteins including TA complexes or isolated toxins/antitoxins. Here, we summarized the structural information of TA systems and analyzed the structural characteristics of known TA modules from several bacteria, especially focusing on the TA modules of several infectious bacteria.

Effects of vertical positions of anterior teeth on smile esthetics in Japanese and korean orthodontists and orthodontic patients.

STATEMENT OF PROBLEM: The perception of a pleasing smile may differ between the countries in Asia or may be converging on a more internationally pleasing one. PURPOSE: The aim of this study was to evaluate the influence of vertical positions of anterior teeth on smile esthetics as assessed by Japanese and Korean orthodontists as well as by orthodontic patients. MATERIALS AND METHODS: A standard composite smile was constructed from different females' intraoral and extraoral photographs. Vertical positions of anterior teeth were modified digitally in 1-mm increments, from 5 mm upper lip coverage of the incisors to 5 mm gingival exposure. Using a visual analog scale, 41 Japanese orthodontists and 96 Japanese orthodontic patients, 25 Korean orthodontists, and 72 Korean orthodontic patients rated the attractiveness of 11 smiles with altered gingival displays. RESULTS: Overall, the Japanese and Korean raters assigned similar esthetic scores between the corresponding groups. The male orthodontic patients assigned higher scores to the smiles with upper lip coverage of the teeth than the corresponding females did in both countries. The female patients assigned clinically significant higher scores to the smiles from 2 mm upper lip coverage of the teeth to 0 mm gingival exposure in both countries. CONCLUSIONS: In both countries, the orthodontists and the female patients shared the similar preference in the smile evaluation of vertical positions of anterior teeth, whereas the male orthodontic patients were more tolerant of upper lip coverage of the anterior teeth than the orthodontists and the female patients. CLINICAL SIGNIFICANCE: This study proposes 0 mm to 2 mm of upper lip coverage of the teeth as a threshold of acceptability for esthetic smile evaluations in female orthodontic patients. For male orthodontic patients, the paper proposes negative gingival display as a threshold of acceptability for esthetic smile evaluations.

Cannabidiol mediates epidermal terminal differentiation and redox homeostasis through aryl hydrocarbon receptor (AhR)-dependent signaling.

BACKGROUND: Cannabidiol, a non-psychoactive phytocannabinoid, has antioxidant and anti-inflammatory activity in keratinocytes. However, the signaling pathway through which cannabidiol exerts its effect on keratinocytes or whether it can modulate keratinocyte differentiation has not been fully elucidated yet. OBJECTIVE: We investigated whether cannabidiol modulates epidermal differentiation and scavenges reactive oxygen species through the aryl hydrocarbon receptor (AhR) in keratinocytes and epidermal equivalents. METHODS: We investigated the cannabidiol-induced activation of AhR using AhR luciferase reporter assay, qRT-PCR, western blot, and immunofluorescence assays. We also analyzed whether keratinocyte differentiation and antioxidant activity are regulated by cannabidiol-induced AhR activation. RESULTS: In both keratinocytes and epidermal equivalents, cannabidiol increased both the mRNA and protein expression of filaggrin, involucrin, NRF2, and NQO1 and the mRNA expression of the AhR target genes, including CYP1A1 and aryl hydrocarbon receptor repressor. Additionally, cannabidiol showed antioxidant activity that was attenuated by AhR knockdown or co-administration with an AhR antagonist. Moreover, cannabidiol increased the ratio of OVOL1/OVOL2 mRNA expression, which is a downstream regulator of AhR that mediates epidermal differentiation. In addition to increased expression of barrier-related proteins, cannabidiol-treated epidermal equivalent showed a more prominent granular layer than the control epidermis. The increased granular layer by cannabidiol was suppressed by the AhR antagonist. CONCLUSION: Cannabidiol can be a modulator of the AhR-OVOL1-filaggrin axis and AhR-NRF2-NQO1 signaling, thus indicating a potential use of cannabidiol in skin barrier enhancement and reducing oxidative stress.

Structure determination of a membrane protein in proteoliposomes.

An NMR method for determining the three-dimensional structures of membrane proteins in proteoliposomes is demonstrated by determining the structure of MerFt, the 60-residue helix-loop-helix integral membrane core of the 81-residue mercury transporter MerF. The method merges elements of oriented sample (OS) solid-state NMR and magic angle spinning (MAS) solid-state NMR techniques to measure orientation restraints relative to a single external axis (the bilayer normal) from individual residues in a uniformly (13)C/(15)N labeled protein in unoriented liquid crystalline phospholipid bilayers. The method relies on the fast (>10(5) Hz) rotational diffusion of membrane proteins in bilayers to average the static chemical shift anisotropy and heteronuclear dipole-dipole coupling powder patterns to axially symmetric powder patterns with reduced frequency spans. The frequency associated with the parallel edge of such motionally averaged powder patterns is exactly the same as that measured from the single line resonance in the spectrum of a stationary sample that is macroscopically aligned parallel to the direction of the applied magnetic field. All data are collected on unoriented samples undergoing MAS. Averaging of the homonuclear (13)C/(13)C dipolar couplings, by MAS of the sample, enables the use of uniformly (13)C/(15)N labeled proteins, which provides enhanced sensitivity through direct (13)C detection as well as the use of multidimensional MAS solid-state NMR methods for resolving and assigning resonances. The unique feature of this method is the measurement of orientation restraints that enable the protein structure and orientation to be determined in unoriented proteoliposomes.