Modulating Phase Behavior in Fatty Acid-Modified Elastin-like Polypeptides (FAMEs): Insights into the Impact of Lipid Length on Thermodynamics and Kinetics of Phase Separation.

Although post-translational lipidation is prevalent in eukaryotes, its impact on the liquid-liquid phase separation of disordered proteins is still poorly understood. Here, we examined the thermodynamic phase boundaries and kinetics of aqueous two-phase system (ATPS) formation for a library of elastin-like polypeptides modified with saturated fatty acids of different chain lengths. By systematically altering the physicochemical properties of the attached lipids, we were able to correlate the molecular properties of lipids to changes in the thermodynamic phase boundaries and the kinetic stability of droplets formed by these proteins. We discovered that increasing the chain length lowers the phase separation temperature in a sigmoidal manner due to alterations in the unfavorable interactions between protein and water and changes in the entropy of phase separation. Our kinetic studies unveiled remarkable sensitivity to lipid length, which we propose is due to the temperature-dependent interactions between lipids and the protein. Strikingly, we found that the addition of just a single methylene group is sufficient to allow tuning of these interactions as a function of temperature, with proteins modified with C7-C9 lipids exhibiting non-Arrhenius dependence in their phase separation, a behavior that is absent for both shorter and longer fatty acids. This work advances our theoretical understanding of protein-lipid interactions and opens avenues for the rational design of lipidated proteins in biomedical paradigms, where precise control over the phase separation is pivotal.

Antibody agonists trigger immune receptor signaling through local exclusion of receptor-type protein tyrosine phosphatases.

Antibodies can block immune receptor engagement or trigger the receptor machinery to initiate signaling. We hypothesized that antibody agonists trigger signaling by sterically excluding large receptor-type protein tyrosine phosphatases (RPTPs) such as CD45 from sites of receptor engagement. An agonist targeting the costimulatory receptor CD28 produced signals that depended on antibody immobilization and were sensitive to the sizes of the receptor, the RPTPs, and the antibody itself. Although both the agonist and a non-agonistic anti-CD28 antibody locally excluded CD45, the agonistic antibody was more effective. An anti-PD-1 antibody that bound membrane proximally excluded CD45, triggered Src homology 2 domain-containing phosphatase 2 recruitment, and suppressed systemic lupus erythematosus and delayed-type hypersensitivity in experimental models. Paradoxically, nivolumab and pembrolizumab, anti-PD-1-blocking antibodies used clinically, also excluded CD45 and were agonistic in certain settings. Reducing these agonistic effects using antibody engineering improved PD-1 blockade. These findings establish a framework for developing new and improved therapies for autoimmunity and cancer.

Dentin remineralization using a stimuli-responsive engineered small molecule GSK3 antagonists-functionalized adhesive.

OBJECTIVES: Tideglusib has shown great performance in terms of dentin regenerative properties. This study aims to evaluate bonding ability, of demineralized dentin infiltrated with polymeric nanoparticles (NPs) doped with tideglusib (TG) (TG-NPs). METHODS: Dentin conditioned surfaces were infiltrated with NPs and TG-NPs. Bonded interfaces were created and stored for 24 h and then submitted to mechanical, chemical and thermal challenging. The resin-dentin interface was evaluated through a doubled dye fluorescent technique and a calcium chelator fluorophore under a confocal laser scanning microscopy, and by field emission scanning electron microscopy. RESULTS: Dentin surfaces treated with TG-NPs and load cycled produced higher bond strength than the rest of the groups. Immersion of dentin specimens treated with undoped-NPs in collagenase solution attained the lowest microtensile bond strength (MTBS) values. Both porosity and nanoleakage decreased when dentin was infiltrated with TG-NPs, that revealed strong signals of xylenol orange stain at both hybrid layer and dentinal tubules. The presence of NPs, in general, inducted the presence of mineralized interfaces after mechanical loading and thermocycling. CONCLUSIONS: Nanoparticles doped with tideglusib promoted the highest dentin bonding efficacy among groups, as they facilitated the maximum bond strength values with creation of mineral deposits at the hybrid layer and dentinal walls. Tideglusib enabled scarce porosity, nanoleakage and advanced sealing among dentin groups. SIGNIFICANCE: Doping hydrophilic polymeric NPs with tideglusib, infiltrated in etched dentin represents a reproducible technique to create reparative dentin at the resin-dentin interface, by inducing therapeutic bioactivity.

Calvaria defect regeneration via human periodontal ligament stem cells and prevascularized scaffolds in athymic rats.

BACKGROUND: Vascularization plays an important role in dental and craniofacial regenerations. Human periodontal ligament stem cells (hPDLSCs) are a promising cell source and, when co-cultured with human umbilical vein endothelial cells (hUVECs), could promote vascularization. The objectives of this study were to develop a novel prevascularized hPDLSC-hUVEC-calcium phosphate construct, and investigate the osteogenic and angiogenic efficacy of this construct with human platelet lysate (hPL) in cranial defects in rats for the first time. METHODS: hPDLSCs and hUVECs were co-cultured on calcium phosphate cement (CPC) scaffolds with hPL. Cell proliferation, angiogenic gene expression, angiogenesis, alkaline phosphatase activity, and cell-synthesized minerals were determined. Bone and vascular regenerations were investigated in rat critical-sized cranial defects in vivo. RESULTS: hPDLSC-hUVEC-CPC-hPL group had 2-fold greater angiogenic expressions and cell-synthesized mineral synthesis than hPDLSC-hUVEC-CPC group (p < 0.05). Microcapillary-like structures were formed on scaffolds in vitro. hPDLSC-hUVEC-CPC-hPL group had more vessels than hPDLSC-hUVEC-CPC group (p < 0.05). In cranial defects in rats, hPDLSC-hUVEC-CPC-hPL group regenerated new bone amount that was 2.1 folds and 4.0 folds, respectively, that of hPDLSC-hUVEC-CPC group and CPC control (p < 0.05). New blood vessel density of hPDLSC-hUVEC-CPC-hPL group was 2 folds and 7.9 folds, respectively, that of hPDLSC-hUVEC-CPC group and CPC control (p < 0.05). CONCLUSION: The hPL pre-culture method is promising to enhance bone regeneration via prevascularized CPC. Novel hPDLSC-hUVEC-CPC-hPL prevascularized construct increased new bone formation and blood vessel density by 4-8 folds over CPC control. CLINICAL SIGNIFICANCE: Novel hPDLSC-hUVEC-hPL-CPC prevascularized construct greatly increased bone and vascular regeneration in vivo and hence is promising for a wide range of craniofacial applications.

Mechanisms of adaptive hypertrophic cardiac remodeling in a large animal model of premature ventricular contraction-induced cardiomyopathy.

Frequent premature ventricular contractions (PVCs) promoted eccentric cardiac hypertrophy and reduced ejection fraction (EF) in a large animal model of PVC-induced cardiomyopathy (PVC-CM), but the molecular mechanisms and markers of this hypertrophic remodeling remain unexplored. Healthy mongrel canines were implanted with pacemakers to deliver bigeminal PVCs (50% burden with 200-220 ms coupling interval). After 12 weeks, left ventricular (LV) free wall samples were studied from PVC-CM and Sham groups. In addition to reduced LV ejection fraction (LVEF), the PVC-CM group showed larger cardiac myocytes without evident ultrastructural alterations compared to the Sham group. Biochemical markers of pathological hypertrophy, such as store-operated Ca2+ entry, calcineurin/NFAT pathway, ß-myosin heavy chain, and skeletal type α-actin were unaltered in the PVC-CM group. In contrast, pro-hypertrophic and antiapoptotic pathways including ERK1/2 and AKT/mTOR were activated and/or overexpressed in the PVC-CM group, which appeared counterbalanced by an overexpression of protein phosphatase 1 and a borderline elevation of the anti-hypertrophic factor atrial natriuretic peptide. Moreover, the potent angiogenic and pro-hypertrophic factor VEGF-A and its receptor VEGFR2 were significantly elevated in the PVC-CM group. In conclusion, a molecular program is in place to keep this structural remodeling associated with frequent PVCs as an adaptive pathological hypertrophy.

Teaching of posterior composites for the restoration of permanent teeth in undergraduate dental training programmes: Systematic review and meta-analysis.

OBJECTIVES: To systematically assess aspects of teaching of posterior composite restorations (PCRs) in permanent teeth in dental schools. STUDY SELECTION: Quantitative studies reporting on dental schools' teaching regarding the placement of PCRs in permanent teeth. Random-effects meta-analyses and meta-regressions were performed. Risk of bias was assessed based on the Medical Education Research Study Quality Instrument (MERSQI). SOURCES: Electronic databases (MEDLINE via PubMed, EMBASE via Ovid, Web of Science, and Scopus) were searched in January 2023. DATA: Forty sources reporting on 34 studies having surveyed 1,286 dental schools were included. Overall, 92.7% (95%-CI: 88.2-95.5) of dental schools reported to teach PCRs. PCRs in three-surface Class II cavities are taught by 82.0% (95%-CI: 70.4-89.7). The mandatory use of liners in deep cavities is taught by 78.3% (95%-CI: 68.9-85.5), and 44.0% (95%-CI: 34.3-54.2) reported to teach bulk-fill composites. While most posterior restorations placed by students were composites (56.1%; 95%-CI: 46.0-65.8), 94.7% (95%-CI: 86.6-98.0) of dental schools (still) teach posterior amalgam restorations. The proportion of dental schools teaching PCRs in three-surface Class II cavities increased and the mean proportion of PCRs among all posterior restorations increased over time (padj.≤0.003). CONCLUSIONS: The teaching of PCRs in dental schools around the world reflects the increased use of resin composite in clinical practice, with students in countries where dental amalgam continues to be used, placing more posterior composites than restorations of dental amalgam. The teaching of PCRs, which is anticipated to increase, will continue to be refined with further developments in adhesive materials, devices, instrumentation, and techniques. CLINICAL SIGNIFICANCE: Graduating dentists can be expected to be familiar with the use of resin composites for the restoration of posterior teeth.

Doped Electrospinned Material-Guides High Efficiency Regional Bone Regeneration.

The main target of bone tissue engineering is to design biomaterials that support bone regeneration and vascularization. Nanostructured membranes of (MMA)1-co-(HEMA)1/(MA)3-co-(HEA)2 loaded with 5% wt of SiO2-nanoparticles (Si-M) were doped with zinc (Zn-Si-M) or doxycycline (Dox-Si-M). Critical bone defects were effectuated on six New Zealand-bred rabbit skulls and then they were covered with the membranes. After six weeks, a histological analysis (toluidine blue technique) was employed to determine bone cell population as osteoblasts, osteoclasts, osteocytes, M1 and M2 macrophages and vasculature. Membranes covering the bone defect determined a higher count of bone cells and blood vessels than in the sham group at the top regions of the defect. Pro-inflammatory M1 appeared in a higher number in the top regions than in the bottom regions, when Si-M and Dox-Si-M were used. Samples treated with Dox-Si-M showed a higher amount of anti-inflammatory and pro-regenerative M2 macrophages. The M1/M2 ratio obtained its lowest value in the absence of membranes. On the top regions, osteoblasts were more abundant when using Si-M and Zn-Si-M. Osteoclasts were equally distributed at the central and lateral regions. The sham group and samples treated with Zn-Si-M attained a higher number of osteocytes at the top regions. A preferential osteoconductive, osteoinductive and angiogenic clinical environment was created in the vicinity of the membrane placed on critical bone defects.

'Thermo-mechanical behavior of alternative material combinations for full-arch implant-supported hybrid prostheses with short cantilevers'.

OBJECTIVES: To compare the fracture resistance (FR) of three combinations of materials for full-arch maxillary implant-supported hybrid prostheses (HPs) with short cantilevers (≤ 10 mm). METHODS: Maxillary HPs were fabricated and classified as follows (n = 5 each): Group-1 (CC-A, control): acrylic-resin-veneered Co-Cr frameworks; Group-2 (CF-A): acrylic-resin-veneered carbon-fiber mesostructures; and Group-3 (CF-R): composite-resin-veneered carbon-fiber frames. Specimens were thermal-cycled (5,000 cycles; 5 °C-55 °C; dwell time: 30 s). Vertical loads were applied until failure, first at the 10-mm-long cantilever (LC), and, afterwards, at the anterior region (AR), using a universal testing machine (crosshead speed: 0.05 mm/s). The fracture pattern was assessed by stereomicroscope and SEM. The one-way ANOVA, the Bonferroni, and the independent samples t tests, were run (α= 0.05). RESULTS: At LC, CF-A, and CC-A samples exhibited the highest FR values (p< 0.001), showing no differences to each other. At AR, CC-A specimens recorded the highest FR, followed by CF-A samples (p< 0.001). CF-R HPs displayed the lowest FR at both locations (p< 0.001). The only group with differences between the tested sites was the CC-A, the AR being more resistant (p< 0.001). Most CC-A and CF-A HPs failed cohesively. CF-R prostheses mainly failed adhesively. CONCLUSIONS: Maxillary HPs with short cantilevers (≤ 10 mm) made of Co-Cr or carbon-fiber veneered with acrylic resin demonstrated an adequate mechanical resistance (> 900 N). CLINICAL SIGNIFICANCE: For maxillary HPs with cantilevers up to 10 mm, acrylic-veneered carbon- fiber mesostructures may be recommended, whereas coating carbon-fiber frames with composite resin seems not suitable.

Teaching of composite restoration repair in Brazilian dental schools.

OBJECTIVES: The aim was to investigate aspects of the teaching of restoration repair as a minimally invasive alternative to replacing defective direct composite restorations in undergraduate curricula teaching programs in Brazilian dental schools. METHODS: A 14-item validated survey questionnaire was mailed to directors/coordinators of operative/restorative dentistry teachers of Brazilian Dental Schools. Data were collected on demographic characteristics of the teachers and institutions, together with questions on the teaching of the repair of defective resin-based composite restorations as part of the school curriculum; the rationale behind the teaching; the nature of the teaching (preclinical and/or clinical); how techniques were taught, indications for repair, operative techniques, materials used, patient acceptability and expected longevity of completed repairs. RESULTS: Two hundred and twenty-two (94%) directors/ coordinators of dental curricula in Brazil were contacted. One hundred and thirty-one directors/coordinators (59%) replied, providing the e-mail address from the teacher responsible for the operative/restorative dentistry program in their school. Of these, 104 responded to the questionnaire (79% response rate). Ninety-three (89%) of the participating schools reported teaching composite repairs as an alternative to replacing restorations. Of the theoretical content, 43% was taught at preclinical and clinical levels, whereas most practical experience (53%) was acquired at clinical levels. Eighty-eight schools (95%) reported tooth substance preservation being the main reason for teaching repair techniques. All schools that taught repairs reported high patient acceptability. CONCLUSIONS: The teaching of composite restoration repair as an alternative to restoration replacement is established in undergraduate programs in most of the Brazilian dental schools surveyed. CLINICAL SIGNIFICANCE: The reasons for teaching restoration repair in Brazil were found to be quite unanimous among teachers, especially regarding the preservation of tooth structure. Variations were found in the clinical indications for repair, suggesting the need for further investigations. Monitoring repaired restorations should be encouraged and could contribute to future studies.

Alterations of sarcoplasmic reticulum-mediated Ca2+ uptake in a model of premature ventricular contraction (PVC)-induced cardiomyopathy.

Premature ventricular contractions (PVCs) are the most frequent ventricular arrhythmias in the overall population. PVCs are known to acutely enhance contractility by the post-extrasystolic potentiation phenomenon, but over time persistent PVCs promote PVC-induced cardiomyopathy (PVC-CM), characterized by a reduction of the left ventricular (LV) ejection fraction. Ca2+ cycling in myocytes commands muscle contraction and in this process, SERCA2 leads the Ca2+ reuptake into the sarcoplasmic reticulum (SR) shaping cytosolic Ca2+ signal decay and muscle relaxation. Altered Ca2+ reuptake can contribute to the contractile dysfunction observed in PVC-CM. To better understand Ca2+ handling using our PVC-CM model (canines with 50% PVC burden for 12 weeks), SR-Ca2+ reuptake was investigated by measuring Ca2+ dynamics and analyzing protein expression. Kinetic analysis of Ca2+ reuptake in electrically paced myocytes showed a ~ 21 ms delay in PVC-CM compared to Sham in intact isolated myocytes, along with a ~ 13% reduction in SERCA2 activity assessed in permeabilized myocytes. Although these trends were not statistically significant between groups using hierarchical statistics, relaxation of myocytes following contraction was significantly slower in PVC-CM vs Sham myocytes. Western blot analyses indicate a 22% reduction in SERCA2 expression, a 23% increase in phospholamban (PLN) expression, and a 50% reduction in PLN phosphorylation in PVC-CM samples vs Sham. Computational analysis simulating a 20% decrease in SR-Ca2+ reuptake resulted in a ~ 22 ms delay in Ca2+ signal decay, consistent with the experimental result described above. In conclusion, SERCA2 and PLB alterations described above have a modest contribution to functional adaptations observed in PVC-CM.

Dexamethasone and zinc loaded polymeric nanoparticles reinforce and remineralize coronal dentin. A morpho-histological and dynamic-biomechanical study.

OBJECTIVE: To investigate the effect of novel polymeric nanoparticles (NPs) doped with dexamethasone (Dex) on viscoelasticity, crystallinity and ultra-nanostructure of the formed hydroxyapatite after NPs dentin infiltration. METHODS: Undoped-NPs, Dex-doped NPs (Dex-NPs) and zinc-doped-Dex-NPs (Zn-Dex-NPs) were tested at dentin, after 24 h and 21 d. A control group without NPs was included. Coronal dentin surfaces were studied by nano-dynamic mechanical analysis measurements, atomic force microscopy, X-ray diffraction and transmission electron microscopy. Mean and standard deviation were analyzed by ANOVA and Student-Newman-Keuls multiple comparisons (p < 0.05). RESULTS: At 21 d of storage time, both groups doped with Dex exhibited the highest complex, storage and loss moduli among groups. Zn-Dex-NPs and Dex-NPs promoted the highest and lowest tan delta values, respectively. Dex-NPs contributed to increase the fibril diameters of dentin collagen over time. Dentin surfaces treated with Zn-Dex-NPs attained the lowest nano-roughness values, provoked the highest crystallinity, and produced the longest and shortest crystallite and grain size. These new crystals organized with randomly oriented lattices. Dex-NPs induced the highest microstrain. Crystalline and amorphous matter was present in the mineral precipitates of all groups, but Zn and Dex loaded NPs helped to increase crystallinity. SIGNIFICANCE: Dentin treated with Zn-Dex-NPs improved crystallographic and atomic order, providing structural stability, high mechanical performance and tissue maturation. Amorphous content was also present, so high hydroxyapatite solubility, bioactivity and remineralizing activity due to the high ion-rich environment took place in the infiltrated dentin.

Novel rechargeable nano-calcium phosphate and nano-calcium fluoride resin cements.

OBJECTIVE: In most clinical circumstances, secondary caries at the margin of fixed dental restorations leads to restoration failure and replacement. Accordingly, the objectives of this study were to: (1) develop a novel rechargeable nano-calcium phosphate (NACP) and nano-calcium fluoride (nCaF2) resin-based cement; and (2) investigate their mechanical properties and calcium (Ca), phosphate (P), and fluoride (F) ion release, recharge, and re-release for the first time. METHODS: The cement matrix consisted of pyromellitic glycerol dimethacrylate (PMGDM), ethoxylated bisphenol-A-dimethacrylate (EBPADMA) was denoted PEHB. Four cements were fabricated: (1) PEHB+0%NACP+0%nCaF2 (experimental control); (2) PEHB+25%NACP+0%nCaF2, (3) PEHB+0%NACP+25%nCaF2; (4) PEHB+12.5%NACP+12.5% nCaF2. RelyX luting cement was used as a commercial control. Mechanical properties and long-term Ca, P, and F ion release, recharge, and re-release were evaluated. RESULTS: Adding 25% NACP, 25% nCaF2 and adding both 12.5% NACP and 12.5% nCaF2 to the cement matrix presented a significantly higher shear bond strength, flexural strength compared to the commercial control (p < 0.05) with a comparable outcome with no significant different (p > 0.05) compared to experimental control. The film thickness results of all cement groups met the ISO requirement (<50 µm). The resin cement group with both 12.5% NACP and 12.5% nCaF2 successfully released Ca, P, and F ions at 3.1 ± 0.01, 1.1 ± 0.05, and 0.51±0.01 mmol/L respectively. Moreover, it showed the ability to re-release Ca, P, and F ions at 0.62±0.01, 0.12±0.01, and 0.42±0.01 mmol/L respectively. CONCLUSIONS: The resin cement group with both 12.5% NACP and 12.5% nCaF2 demonstrated the advantages of both types of bio-interactive fillers as it could release a higher level of ions than the resin cement with 25%nCAF2 and exhibited a better rechargeability compared to the resin cement with 25%NACP. CLINICAL SIGNIFICANCE: The ability of this novel resin-based cement to release, recharge, and re-release Ca, P, and F ions could be one of the keys to lengthening the survivability of fixed dental restorations. These features could help to reduce the onset of secondary caries by enhancing the remineralization and preventing the demineralization of tooth structures.

Response rates to questionnaire-based studies in the contemporary dental literature: A systematic review.

OBJECTIVES: This systematic review aimed to investigate what is a reasonable response rate for dental questionnaire-based studies in recent literature and to assess the factors that affect the response rates. METHODS: We used MEDLINE/PubMed to search the dental literature of 2021 (January-October). Two reviewers independently assessed studies eligibility and extracted data using standardized electronic extraction form. RESULTS: One hundred and seventy-two studies were eligible, of these a total of 149 response rates were reported from 133 studies, whereas the remaining 39 studies were excluded as they did not report response rates. The median response rate across the included studies was 77% (mean = 70.8%). We found significant negative correlation between the response rate and the actual number of distributed questionnaires (sample size) (r = -0.4127; P < 0.001). We also found an association between the response rate and the area of distribution, e.g., national or international (P = 0.0012). However, a wide variation was observed in the quality of information reported within this review and we did not find clear evidence of association between the response rate and other variables such as questionnaire piloting, number of questions in the questionnaire and the journal impact factor. CONCLUSIONS: The findings of this systematic review confirm the association between the response rate and the sample size, where the response rate increases when the sample size less than 300 participants. In addition, a higher response rate could be achieved when the study conducted within the same institution (e.g., university). SIGNIFICANCE: Questionnaire-based research can provide answers to several questions that could not be answered by other types of research related to the field of dentistry, dental health practitioners and students' attitudes and behaviours and more. Questionnaire-based publications can effectively contribute to dental research; thus, dental journals should consider development of a minimum set of guidelines in the reporting of questionnaire-based manuscripts.

Estimating the Health Effects of Adding Bicycle and Pedestrian Paths at the Census Tract Level: Multiple Model Comparison.

BACKGROUND: Adding additional bicycle and pedestrian paths to an area can lead to improved health outcomes for residents over time. However, quantitatively determining which areas benefit more from bicycle and pedestrian paths, how many miles of bicycle and pedestrian paths are needed, and the health outcomes that may be most improved remain open questions. OBJECTIVE: Our work provides and evaluates a methodology that offers actionable insight for city-level planners, public health officials, and decision makers tasked with the question "To what extent will adding specified bicycle and pedestrian path mileage to a census tract improve residents' health outcomes over time?" METHODS: We conducted a factor analysis of data from the American Community Survey, Center for Disease Control 500 Cities project, Strava, and bicycle and pedestrian path location and use data from two different cities (Norfolk, Virginia, and San Francisco, California). We constructed 2 city-specific factor models and used an algorithm to predict the expected mean improvement that a specified number of bicycle and pedestrian path miles contributes to the identified health outcomes. RESULTS: We show that given a factor model constructed from data from 2011 to 2015, the number of additional bicycle and pedestrian path miles in 2016, and a specific census tract, our models forecast health outcome improvements in 2020 more accurately than 2 alternative approaches for both Norfolk, Virginia, and San Francisco, California. Furthermore, for each city, we show that the additional accuracy is a statistically significant improvement (P<.001 in every case) when compared with the alternate approaches. For Norfolk, Virginia (n=31 census tracts), our approach estimated, on average, the percentage of individuals with high blood pressure in the census tract within 1.49% (SD 0.85%), the percentage of individuals with diabetes in the census tract within 1.63% (SD 0.59%), and the percentage of individuals who had >2 weeks of poor physical health days in the census tract within 1.83% (SD 0.57%). For San Francisco (n=49 census tracts), our approach estimates, on average, that the percentage of individuals who had a stroke in the census tract is within 1.81% (SD 0.52%), and the percentage of individuals with diabetes in the census tract is within 1.26% (SD 0.91%). CONCLUSIONS: We propose and evaluate a methodology to enable decision makers to weigh the extent to which 2 bicycle and pedestrian paths of equal cost, which were proposed in different census tracts, improve residents' health outcomes; identify areas where bicycle and pedestrian paths are unlikely to be effective interventions and other strategies should be used; and quantify the minimum amount of additional bicycle path miles needed to maximize health outcome improvements. Our methodology shows statistically significant improvements, compared with alternative approaches, in historical accuracy for 2 large cities (for 2016) within different geographic areas and with different demographics.

Effects of stimulus waveform on transcranial magnetic stimulation metrics in proximal and distal arm muscles.

OBJECTIVES: The purpose of this study was to determine the effect of common transcranial magnetic stimulation (TMS) waveforms (monophasic and biphasic) on resting motor threshold (RMT), active motor threshold (AMT), and motor evoked potential (MEP) amplitudes in the biceps and first dorsal interosseous (FDI) because waveforms may affect motor targets differently. We also determined test-retest reliability. METHODS: Ten individuals participated in two sessions of TMS delivered to the motor cortex. Monophasic stimulation to induce a posterior-anterior current in the brain (monoPA) and biphasic posterior-anterior then anterior-posterior (biPA-AP) were applied in each session in random order. In each session, there were four blocks of measurements (2 muscles × 2 waveforms) of RMT, AMT and MEPs at the hotspot location. MEPs were normalized to the maximum EMG signal. RESULTS: RMTs and AMTs were lower for monoPA compared to biPA-AP stimulation for the biceps (p<0.01) and FDI (p<0.01). Normalized MEPs were greater for monoPA compared to biPA-AP stimulation in the FDI (p=0.01) and not different in the biceps (p=0.86). Motor thresholds were not different between sessions suggesting high reliability (p<0.01). Normalized MEPs had very low reliability across sessions in the FDI, and moderate reliability in the biceps. DISCUSSION: Preliminary investigation suggests the effect of TMS waveform on motor thresholds is similar in upper limb proximal and distal muscles, but the effect differs per motor target for MEPs. Further, test-retest reliability of waveform effects was sensitive to target muscle. These findings may contribute to improve the efficacy and reliability of TMS for clinical use.

Human periodontal ligament stem cell encapsulation in alginate-fibrin-platelet lysate microbeads for dental and craniofacial regeneration.

OBJECTIVE: Tissue engineering is promising for dental and craniofacial regeneration. The objectives of this study were to develop a novel xeno-free alginate-fibrin-platelet lysate hydrogel with human periodontal ligament stem cells (hPDLSCs) for dental regeneration, and to investigate the proliferation and osteogenic differentiation of hPDLSCs using hPL as a cell culture nutrient supplement. METHODS: hPDLSCs were cultured with Dulbecco's modified eagle medium (DMEM), DMEM + 10% fetal bovine serum (FBS), and DMEM + hPL (1%, 2.5%, and 5%). hPDLSCs were encapsulated in alginate-fibrin microbeads (Alg+Fib), alginate-hPL microbeads (Alg+hPL), or alginate-fibrin-hPL microbeads (Alg+Fib+hPL). hPDLSCs encapsulated in alginate microbeads were induced with an osteogenic medium containing hPL or FBS. Quantitative real-time polymerase chain reaction (qRT-PCR), alkaline phosphatase (ALP) activity, ALP staining, and alizarin red (ARS) staining was investigated. RESULTS: hPDLSCs were released faster from Alg+Fib+hPL than from Alg+hPL. At 14 days, ALP activity was 44.1 ± 7.61 mU/mg for Alg+Fib+hPL group, higher than 28.07 ± 5.15 mU/mg of Alg+Fib (p<0.05) and 0.95 ± 0.2 mU/mg of control (p<0.01). At 7 days, osteogenic genes (ALP, RUNX2, COL1, and OPN) in Alg+Fib+hPL and Alg+Fib were 3-10 folds those of control. At 21 days, the hPDLSC-synthesized bone mineral amount in Alg+Fib+hPL and Alg+Fib was 7.5 folds and 4.3 folds that of control group, respectively. CONCLUSIONS: The 2.5% hPL was determined to be optimal for hPDLSCs. Adding hPL into alginate hydrogel improved the viability of the hPDLSCs encapsulated in the microbeads. The hPL-based medium enhanced the osteogenic differentiation of hPDLSCs in Alg+Fib+hPL construct, showing a promising xeno-free approach for delivering hPDLSCs to enhance dental, craniofacial and orthopedic regenerations.

Effects of thermal cycling on mechanical and antibacterial durability of bioactive low-shrinkage-stress nanocomposite.

OBJECTIVES: Recent studies developed low-shrinkage-stress composite with remineralizing and antibacterial properties to combat secondary caries and increase restoration longevity. However, their long-term durability in thermal cycling is unclear. The objectives of this study were to develop an antibacterial, remineralizing and low-shrinkage-stress composite, and to investigate its durability in thermal cycling for 20,000 cycles, equivalent to two years of clinical life. METHODS: The resin consisted of urethane dimethacrylate (UDMA) and triethylene glycol divinylbenzyl ether (TEG-DVBE). Composites were made with 5% dimethylaminohexadecyl methacrylate (DMAHDM) and 20% of nanoparticles of amorphous calcium phosphate (NACP). Composites were thermal cycled at 5°C and 55°C for 20,000 cycles. A human salivary biofilm model was used to evaluate antibiofilm activity before and after thermal cycling. RESULTS: After 20,000 cycles, the flexural strength of bioactive low-shrinkage-stress composite matched commercial control with no antibacterial activity (p > 0.05). Surface roughness was clinically acceptable at less than 0.2 µm. UV+NACP+DMAHDM composite reduced the total microorganisms, total streptococci, and mutans streptococci by 2-5 logs, compared to commercial composite. Biofilm lactic acid production was reduced by 11 folds. The antibacterial performance was maintained after thermal cycling, with no decrease after 20,000 cycles. CONCLUSIONS: Bioactive low-shrinkage-stress composite possessed good mechanical properties that matched commercial composite both before and after thermal cycling. The new composite had potent antibacterial activity, which was maintained and did not decrease after thermal cycling. CLINICAL SIGNIFICANCE: The new bioactive low-shrinkage-stress composite could reduce polymerization shrinkage stress and release calcium and phosphate ions, with good mechanical properties and strong antibacterial function that were durable after thermal cycling. These properties indicate great potential for inhibiting recurrent caries and increasing the restoration longevity.

Current and future trends in the teaching of removable partial dentures in dental schools in Malaysia: A cross sectional study.

AIMS: To investigate, using a validated questionnaire, the teaching of removable partial dentures (RPDs) in dental schools of Malaysia. MATERIALS AND METHODS: A validated questionnaire to investigating trends in the teaching of RPDs in dental schools in Oceania was emailed (in English version form) to Heads of Restorative/Prosthodontics/course coordinators in the 13 dental schools in Malaysia. Follow-up reminders were sent and participants were given six weeks to complete and return the questionnaire. Data was entered into an Excel spreadsheet and results compiled and analyzed. RESULTS: Completed questionnaires were received from 13 dental school - a 100% response rate. All schools (n = 13) provided a preclinical technical course in RPD design. In most schools (n = 9, 69.2%), course work was supervised by senior lecturers while rest of the institutions made use of associate professor/professors. There were significant differences (p<0.05) between dental schools in terms of the contact hours dedicated to preclinical teaching. Students received an average of 62 h of instruction. Didactic instruction was the primary focus with practical (78 h) and didactic teaching (32 h). All dental schools (n = 13) provided practical surveyor design teaching (8 h). The staff student ratio for formal lectures (1:61), tutorials (1:29) and lab demonstrations (1:12) were recorded. Majority of the schools (n = 11, 84.6%) employed paired teaching in clinical sessions. All schools (n = 13, 100%) emphasized on increased teaching of RPD design and prescription writing in future clinical RPD courses. CONCLUSION: Teaching of RPDs in Malaysia may be considered sufficient and comparable to the teaching in other parts of the world. CLINICAL SIGNIFICANCE: Whilst the teaching of partial dentures at Dental Schools in Malaysia provides the core competencies involved in partial denture design and construction based on sound fundamental, scientific principles they should address the challenges of teaching partial dentures and other areas of dental education including improving working conditions for dental professionals.