Association between triglycerides, known risk SNVs and conserved rare variation in SLC25A40 in a multi-ancestry cohort.

BACKGROUND: Elevated triglycerides (TG) are associated with, and may be causal for, cardiovascular disease (CVD), and co-morbidities such as type II diabetes and metabolic syndrome. Pathogenic variants in APOA5 and APOC3 as well as risk SNVs in other genes [APOE (rs429358, rs7412), APOA1/C3/A4/A5 gene cluster (rs964184), INSR (rs7248104), CETP (rs7205804), GCKR (rs1260326)] have been shown to affect TG levels. Knowledge of genetic causes for elevated TG may lead to early intervention and targeted treatment for CVD. We previously identified linkage and association of a rare, highly conserved missense variant in SLC25A40, rs762174003, with hypertriglyceridemia (HTG) in a single large family, and replicated this association with rare, highly conserved missense variants in a European American and African American sample. METHODS: Here, we analyzed a longitudinal mixed-ancestry cohort (European, African and Asian ancestry, N = 8966) from the Electronic Medical Record and Genomics (eMERGE) Network. We tested associations between median TG and the genes of interest, using linear regression, adjusting for sex, median age, median BMI, and the first two principal components of ancestry. RESULTS: We replicated the association between TG and APOC3, APOA5, and risk variation at APOE, APOA1/C3/A4/A5 gene cluster, and GCKR. We failed to replicate the association between rare, highly conserved variation at SLC25A40 and TG, as well as for risk variation at INSR and CETP. CONCLUSIONS: Analysis using data from electronic health records presents challenges that need to be overcome. Although large amounts of genotype data is becoming increasingly accessible, usable phenotype data can be challenging to obtain. We were able to replicate known, strong associations, but were unable to replicate moderate associations due to the limited sample size and missing drug information.

Sulfated tyrosines 27 and 29 in the N-terminus of human CXCR3 participate in binding native IP-10.

AIM: Human CXCR3, a seven-transmembrane segment (7TMS), is predominantly expressed in Th1-mediated responses. Interferon-gamma-inducible protein 10 (IP-10) is an important ligand for CXCR3. Their interaction is pivotal for leukocyte migration and activation. Tyrosine sulfation in 7TMS is a posttranslational modification that contributes substantially to ligand binding. We aimed to study the role of tyrosine sulfation of CXCR3 in the protein's binding to IP-10. METHODS: Plasmids encoding CXCR3 and its mutants were prepared by PCR and site-directed mutagenesis. HEK 293T cells were transfected with plasmids encoding CXCR3 or its variants using calcium phosphate. Transfected cells were labeled with [(35)S]-cysteine and methionine or [(35)S]-Na(2)SO(3) and then analyzed by immunoprecipitation to measure sulfation. Experiments with (125)I-labeled IP-10 were carried out to evaluate the affinity of CXCR3 for its ligand. Calcium influx assays were used to measure intercellular signal transduction. RESULTS: Our data show that sulfate moieties are added to tyrosines 27 and 29 of CXCR3. Mutation of these two tyrosines to phenylalanines substantially decreases binding of CXCR3 to IP-10 and appears to eliminate the associated signal transduction. Tyrosine sulfation of CXCR3 is enhanced by tyrosyl protein sulfotransferases (TPSTs), and it is weakened by shRNA constructs. The binding ability of CXCR3 to IP-10 is increased by TPSTs and decreased by shRNAs. CONCLUSIONS: This study identifies two sulfated tyrosines in the N-terminus of CXCR3 as part of the binding site for IP-10, and it underscores the fact that tyrosine sulfation in the N-termini of 7TMS receptors is functionally important for ligand interactions. Our study suggests a molecular target for inhibiting this ligand-receptor interaction.

[Regulation of tyrosylprotein sulfotransferases activity by sulfotyrosine].

OBJECTIVE: To investigate the role of sulfated tyrosine in regulating the activity of tyrosylprotein sulfotransferases (TPST) 1 and TPST2. METHODS: Constructs of TPST 1 and TPST2 were amplified by polymerase chain reaction (PCR), then fused into immunoglobulin G1 Fc region. All the variants in which sulfated tyrosines were mutated to phenylalanine were made by the PCR-based Quick Change method and confirmed by sequencing the entire reading frame. Small hairpin RNA (shRNA) constructs-targeting nucleotides 259-275 of TPST1 and nucleotides 73-94 of TPST2 were generated and subcloned into pBluescript. Human embryonic kidney (HEK) 293T cells were transfected with these plasmids. One day later, cells were split: one part was labeled with 35S-cysteine and methionine or 35S-Na2SO3 overnight, the second part was used for 125I labeled binding experiment, and the third part was retained for binding and flow cytometry. RESULTS: Tyrosines at position 326 of TPST1 and position 325 of TPST2 were sulfated posttranslationally. Tyrosine sulfation of TPSTs was effectively inhibited by sulfation inhibitors, including specific shRNAs and non-specific NaCIO3. shRNAs reduced the sulfation of C3a receptor and C5a receptor, and partially blocked the binding of these two receptors to their respective ligands. CONCLUSIONS: The activities of TPSTs were regulated by tyrosine sulfation. Inhibition of sulfotyrosine decreases the binding ability of C3a receptor and C5a receptor to their respective ligands.

[Association analysis of 30 type 2 diabetes candidate genes in Chinese Han population].

OBJECTIVE: To identify the susceptibility genes of type 2 diabetes in Chinese Han population. METHODS: Single nucleotide polymorphism (SNP) discovery, genotyping and haplotype construction were performed in 30 candidate genes. Case-control study were carried out in a population-based sample and confirmed by the transmission disequilibrium test (TDT) analysis in 77 trio pedigrees. The effects of the SNP rs5210 on gene expression were studied by reporter gene technique. RESULTS: The case-control studies showed that several SNPs on KCNJ11 gene was associated with type 2 diabetes in Chinese Han population, in which the allele frequency of SNP rs5219, the genotype frequency of rs5210, rs2285676 and rs5219, and the frequency of haplotype GA combined of the rs5219 and rs5215 showed significant difference between these two groups (P < 0.05). In addition, TDT test also showed statistical significance on this haplotype GA (P < 0. 05). The reporter gene assay showed that the effect on gene expression was significantly different between two alleles of rs5210 (P < 0.05). CONCLUSION: KCNJII gene is one of the susceptibility genes of type 2 diabetes in Chinese Han population.

Bond strength of metal brackets bonded to a silica-based ceramic with light-cured adhesive : Influence of various surface treatment methods.

OBJECTIVE: The purpose of this work was to evaluate the effects of several surface treatment methods on the shear bond strengths of metal brackets bonded to a silica-based ceramic with a light-cured adhesive. MATERIALS AND METHODS: Silica-based ceramic (IPS Classic(®)) with glazed surfaces was cut into discs that were used as substrates. A total of 80 specimens were randomly divided into four groups according to the method used: 9.6 % hydrofluoric acid (group 1), 9.6 % hydrofluoric acid (HF) + silane coupling agent (group 2), sandblasting (aluminum trioxide, 50 µm) + silane (group 3), and tribochemical silica coating (CoJet™ sand, 30 µm) + silane (group 4). Brackets were bonded to the treated specimens with a light-cure adhesive (Transbond XT, 3 M Unitek). Shear bond strength was tested after bracket bonding, and the Adhesive Remnant Index (ARI) scores were quantified after debonding. RESULTS: Group 4 showed the highest bond strength (12.3 ± 1.0 MPa), which was not significantly different from that of group 3 (11.6 ± 1.2 MPa, P > 0.05); however, the bond strength of group 4 was substantially higher than that of group 2 (9.4 ± 1.1 MPa, P < 0.05). The shear bond strength of group 1 (3.1 ± 0.6 MPa, P< 0.05) was significantly lower than that of the other groups. CONCLUSION: Shear bond strengths exceeded the optimal range of ideal bond strength for clinical practice, except for the isolated HF group. HF acid etching followed by silane was the best suited method for bonding on IPS Classic(®). Failure modes in the sandblasting and silica-coating groups revealed signs of damaged ceramic surfaces.

[Nuclear localization region in soluble adenylyl cyclase].

OBJECTIVE: To locate the region responsible for nuclear localization of protein sAC. METHODS: The eukaryotic expression vector of vairous sAC deletion mutants were transfected into Hela cells. The localization of each mutant was observed using confocal microscope. RESULTS: For some mutants, the localization of sAC changed. Deletion of some region made it unable to locate in the nuclear. CONCLUSION: It is possible to figure out that the nucleotide region (739-1038 and 1045-1261) take charge of nuclear localization of sAC.

[Dental arch form reverting by four-point method].

PURPOSE: To explore a simple method of reverting individual dental arch form template for wire bending. METHODS: Individual dental arch form was reverted by four-point method. By defining central point of bracket on bilateral lower second premolar and first molar, certain individual dental arch form could be generated. The arch form generating procedure was then be developed to computer software for printing arch form. Four-point method arch form was evaluated by comparing with direct model measurement on linear and angular parameters. The accuracy and reproducibility were assessed by paired t test and concordance correlation coefficient with Medcalc 9.3 software package. RESULTS: The arch form by four-point method was of good accuracy and reproducibility (linear concordance correlation coefficient was 0.9909 and angular concordance correlation coefficient was 0.8419). CONCLUSION: The dental arch form reverted by four-point method could reproduce the individual dental arch form.

[Effects of different premolar extraction on lower third molar eruption].

PURPOSE: To analyze the effect of different premolar extraction on the position of the mandibular third molar during orthodontic treatment. METHODS: Cephalometric radiographs were taken before and after 2 years of treatment in 28 female patients selected from Department of Orthodontic, Ninth People's Hospital, School of Medicine, Shanghai Jiao Tong University. Treatment for 12 patients in group I included extraction of 4 second premolars,the other 16 patients in group II were treated with extraction of 4 first premolars. Modified sectional arch wire technique was used to redistribute the spaces.Change of the mandibular third molar position was valued by tracing. The data was analyzed using SAS6.12 software package for student's t test. RESULTS: The vertical change in the third molar position in group I and II was (4.58+/-1.85)mm and (0.21+/-1.11)mm (P<0.01), the horizontal change was (2.95+/-2.55)mm and (-0.03+/-1.74)mm (P<0.01), the mesio-angulation was decreased (-10.12+/-8.27) degrees and (-5.06+/-3.60) degrees (P<0.05),and the angle to the second molar decreased (-15.58+/-9.65) degrees and (-4.21+/-3.68) degrees (P<0.001), respectively. CONCLUSION: Second premolar extraction may increase the rates of movement of the third molar in horizontal and vertical direction and decrease the mesio-angulation and the angle to the second molar than first premolar extraction.

[Evaluations of self-perceived orthodontic treatment need using 2 different scales in adolescents].

PURPOSE: To evaluate the self-perceived dental aesthetics through visualized analogue scale (VAS) and aesthetic component (AC) of index of orthodontic treatment (IOTN) among adolescents in Shanghai area. METHODS: The investigation was carried out among 302 students (148 boys and 144 girls) aged 11-13 years. Self-perception of the dental aesthetic appearance was evaluated through VAS and AC. Additionally, the objective dental aesthetics were scored by orthodontists using AC and the actual dental attractiveness satisfaction was determined by a simple question. The data was analyzed using SPSS11.0 software package for Spearman correlation analysis. RESULTS: Generally, no statistically significant sex differences were found in relation to the VAS score and SAC degree (P > 0.05). No correlation was found between actual dental attractiveness satisfaction and self-perceived AC grade (r = 0.04. P = 0.441). However, statistically significant, positive, strong correlations were found between the actual dental attractiveness satisfaction and VAS score (r = 0.80, P = 0.000). And meaningful relation between the AC and VAS score was found. CONCLUSIONS: VAS showed high ability to predict the self-perceived dental aesthetics and act as a simple and useful tool, which can be used in further investigations.