Three-dimensional decision support system for treatment of canine impaction.

INTRODUCTION: This study aimed to develop a 3-dimensional (3D) characterization of the severity of maxillary impacted canines and to test the clinical performance of this characterization as a treatment decision support tool. METHODS: Cone-beam computed tomography images obtained from 83 patients with 120 impacted maxillary canines were included. Quantitative information on the canine 3D position and qualitative assessment of root damage of adjacent teeth were evaluated. A severity index was constructed on the basis of the quantitative findings. Clinical applicability was tested by comparing clinical diagnosis and treatment planning for conventional records vs the 3D characterization via a 2-part survey. RESULTS: The average quantitative assessments of impacted maxillary canine position were 6.4 ± 3.6 mm from the midsagittal plane, 11.6 ± 3.1 mm in height relative to the occlusal plane, 31.5° ± 18° of roll, and 48.8° ± 14.3° of pitch. The severity index ranged from 0-13 with a mean score of 4.5 ± 2.2. Overlap with adjacent teeth was the greatest contributor (33%) to the index. Bicortically impacted canines caused the most severe root damage. Cone-beam computed tomography was preferred for assessing root damage and overall severity, whereas conventional imaging was sufficient for height and angulation assessment. The 3D report was very important or important for evaluating root damage, canine position, overall severity, and overlap. The 3D report changed most of the decisions relating to biomechanics, patient education, and treatment time estimate. The decision of exposure and traction vs extraction was changed 22% of the time after the presentation of the 3D report. CONCLUSIONS: The overlap with adjacent teeth frequently contributes the most to the severity index. The 3D report provided relevant clinical information regarding the canine position, damage to adjacent teeth, and the severity index, with a profound impact on the decisions of the clinicians regarding biomechanics, patient education, and treatment time estimate.

Dental long axes using digital dental models compared to cone-beam computed tomography.

OBJECTIVE: Standard methods of evaluating tooth long axes are not comparable (digital dental models [DDMs], panoramic and cephalometric radiographs) or expose patients to more radiation (cone-beam computed tomography [CBCT]). This study aimed to compare angular changes in tooth long axes using DDMs vs using CBCTs. SETTINGS AND SAMPLE POPULATION: Secondary data analysis of DDMs and CBCTs, taken before and after orthodontic treatment with piezocision of 24 patients. METHODS: Angular changes in tooth long axes were evaluated using landmarks on first molars (centre of the occlusal surface and centre of the furcation), canines and incisors (cusp tip and centre of the root at the cementoenamel junction). Wilcoxon test, intraclass correlation coefficient (ICC) and Bland-Altman plots were used to test intra- and inter-rater agreement and compare DDM and CBCT measurements. RESULTS: The mesiodistal angulation and buccolingual inclination DDM measurements were reproducible. Overall mean differences between DDM and CBCT measurements of mesiodistal angulation, 1.9°±1.5°, and buccolingual inclination, 2.2 ± 2.2°, were not significant for all teeth. ICC between DDM and CBCT measurements ranged from good (0.85 molars) to excellent (0.94 canines; 0.96 incisors). The percentages of measurements outside the range of ±5 were 17.4% for molars, 13.8% for canines and 4.5% for incisors. CONCLUSIONS: DDM assessment of changes in tooth long axes has good reproducibility and yields comparable measurements to those obtained from CBCT within a 5° range. These findings lay the groundwork for machine learning approaches that synthesize crown and root canal information towards planning tooth movement without the need for ionizing radiation scans.

Presurgical orthodontic decompensation with clear aligners.

INTRODUCTION: Orthodontists, surgeons, and patients have taken an interest in using clear aligners in combination with orthognathic surgery. This study aimed to evaluate the accuracy of tooth movements with clear aligners during presurgical orthodontics using novel 3-dimensional superimposition techniques. METHODS: The study sample consisted of 20 patients who have completed presurgical orthodontics using Invisalign clear aligners. Initial (pretreatment) digital dental models, presurgical digital dental models, and ClinCheck prediction models were obtained. Presurgical models were superimposed onto initial ones using stable anatomic landmarks; ClinCheck models were superimposed onto presurgical models using surface best-fit superimposition. Five hundred forty-five teeth were measured for 3 angular movements (buccolingual torque, mesiodistal tip, and rotation) and 4 linear movements (buccolingual, mesiodistal, vertical, and total scalar displacement). The predicted tooth movement was compared with the achieved amount for each movement and tooth, using both percentage accuracy and numerical difference. RESULTS: Average percentage accuracy (63.4% ± 11.5%) was higher than in previously reported literature. The most accurate tooth movements were buccal torque and mesial displacement compared with lingual torque and distal displacement, particularly for mandibular posterior teeth. Clinically significant inaccuracies were found for the buccal displacement of maxillary second molars, lingual displacement of all molars, intrusion of mandibular second molars, the distal tip of molars, second premolars, and mandibular first premolars, buccal torque of maxillary central and lateral incisors, and lingual torque of premolars and molars. CONCLUSIONS: Superimposition techniques used in this study lay the groundwork for future studies to analyze advanced clear aligner patients. Invisalign is a treatment modality that can be considered for presurgical orthodontics-tooth movements involved in arch leveling and decompensation are highly accurate when comparing the simulated and the clinically achieved movements.

Allosteric suppression of HIV-1 reverse transcriptase structural dynamics upon inhibitor binding.

Efavirenz is a second-generation nonnucleoside reverse transcriptase inhibitor (NNRTI) and a common component of clinically approved anti-AIDS regimens. NNRTIs are noncompetitive inhibitors that bind in a hydrophobic pocket in the p66 subunit of reverse transcriptase (RT) ∼10 Å from the polymerase active site. Hydrogen exchange mass spectrometry (HXMS) shows that efavirenz binding reduces molecular flexibility in multiple regions of RT heterodimer in addition to the NNRTI binding site. Of the 47 peptic fragments monitored by HXMS, 15 showed significantly altered H/D exchange rates in the presence of efavirenz. The slow cooperative unfolding of a ß-sheet in the NNRTI binding pocket, which was previously observed in unliganded RT, is dramatically suppressed by efavirenz. HXMS also defines an extensive network of allosterically coupled sites, including four distinct regions of allosteric stabilization, and one region of allosteric destabilization. The effects of efavirenz binding extend > 60 Å from the NNRTI binding pocket. Allosteric changes to the structural dynamics propagate to the thumb and connection subdomains and RNase H domain of the p66 subunit as well as the thumb and palm subdomains of the p51 subunit. These allosteric regions may represent potential new drug targets.

Efavirenz binding to HIV-1 reverse transcriptase monomers and dimers.

Efavirenz (EFV) is a nonnucleoside reverse transcriptase inhibitor (NNRTI) of HIV-1 reverse transcriptase (RT) used for the treatment of AIDS. RT is a heterodimer composed of p66 and p51 subunits; p51 is produced from p66 by C-terminal truncation by HIV protease. The monomers can form p66/p66 and p51/p51 homodimers as well as the p66/p51 heterodimer. Dimerization and efavirenz binding are coupled processes. In the crystal structure of the p66/p51-EFV complex, the drug is bound to the p66 subunit. The binding of efavirenz to wild-type and dimerization-defective RT proteins was studied by equilibrium dialysis, tryptophan fluorescence, and native gel electrophoresis. A 1:1 binding stoichiometry was determined for both monomers and homodimers. Equilibrium dissociation constants are approximately 2.5 microM for both p66- and p51-EFV complexes, 250 nM for the p66/p66-EFV complex, and 7 nM for the p51/p51-EFV complex. An equilibrium dissociation constant of 92 nM for the p66/p51-EFV complex was calculated from the thermodynamic linkage between dimerization and inhibitor binding. Binding and unbinding kinetics monitored by fluorescence were slow. Progress curve analyses revealed a one-step, direct binding mechanism with association rate constants k(1) of approximately 13.5 M(-1) s(-1) for monomers and heterodimer and dissociation rate constants k(-1) of approximately 9 x 10(-5) s(-1) for monomers. A conformational selection mechanism is proposed to account for the slow association rate. These results show that efavirenz is a slow, tight-binding inhibitor capable of binding all forms of RT and suggest that the NNRTI binding site in monomers and dimers is similar.

Efavirenz binding site in HIV-1 reverse transcriptase monomers.

Efavirenz (EFV) is a potent nonnucleoside reverse transcriptase inhibitor (NNRTI) used in the treatment of AIDS. NNRTIs bind in a hydrophobic pocket located in the p66 subunit of reverse transcriptase (RT), which is not present in crystal structures of RT without an inhibitor. Recent studies showed that monomeric forms of the p66 and p51 subunits bind efavirenz with micromolar affinity. The effect of efavirenz on the solution conformations of p66 and p51 monomers was studied by hydrogen-deuterium exchange mass spectrometry (HXMS) and Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). HXMS data reveal that five peptides, four of which contain efavirenz contact residues seen in the crystal structure of the RT-EFV complex, exhibit a reduced level of exchange in monomer-EFV complexes. Moreover, peptide 232-246 undergoes slow cooperative unfolding-refolding in the bound monomers, but at a rate much slower than that observed in the p66 subunit of the RT heterodimer [Seckler, J. M., Howard, K. J., Barkley, M. D., and Wintrode, P. L. (2009) Biochemistry 48, 7646-7655]. These results suggest that the efavirenz binding site on p66 and p51 monomers is similar to the NNRTI binding pocket in the p66 subunit of RT. Nanoelectrospray ionization FT-ICR mass spectra indicate that the intact monomers each have (at least) two different conformations. In the presence of efavirenz, the mass spectra change significantly and suggest that p51 adopts a single, more compact conformation, whereas p66 undergoes facile, electrospray-induced cleavage. The population shift is consistent with a selected-fit binding mechanism.

Solution structural dynamics of HIV-1 reverse transcriptase heterodimer.

Crystal structures and simulations suggest that conformational changes are critical for the function of HIV-1 reverse transcriptase. The enzyme is an asymmetric heterodimer of two subunits, p66 and p51. The two subunits have the same N-terminal sequence, with the p51 subunit lacking the C-terminal RNase H domain. We used hydrogen exchange mass spectrometry to probe the structural dynamics of RT. H/D exchange revealed that the fingers and palm subdomains of both subunits form the stable core of the heterodimer. In the crystal structure, the tertiary fold of the p51 subunit is more compact than that of the polymerase domain of the p66 subunit, yet both subunits show similar flexibility. The p66 subunit contains the polymerase and RNase H catalytic sites. H/D exchange indicated that the RNase H domain of p66 is very flexible. The beta-sheet beta12-beta13-beta14 lies at the base of the thumb subdomain of p66 and contains highly conserved residues involved in template/primer binding and NNRTI binding. Using the unique ability of hydrogen exchange mass spectrometry to resolve slowly interconverting species, we found that beta-sheet beta12-beta13-beta14 undergoes slow cooperative unfolding with a t(1/2) of <20 s. The H/D exchange results are discussed in relation to existing structural, simulation, and sequence information.

Kinetics of association and dissociation of HIV-1 reverse transcriptase subunits.

The biologically active form of HIV-1 reverse transcriptase (RT) is the p66/p51 heterodimer. The process of maturation of the heterodimer from precursor proteins is poorly understood. Previous studies indicated that association of p66 and p51 is very slow. Three techniques, a pre-steady-state activity assay, intrinsic tryptophan fluorescence, and a FRET assay, were used to monitor the dimerization kinetics of RT. Kinetic experiments were conducted with purified p66 and p51 proteins in aqueous buffer. All three techniques gave essentially the same results. The dissociation kinetics of p66/p51 were first-order with rate constants (k(diss)) of approximately 4 x 10(-6) s(-1) (t(1/2) = 48 h). The association kinetics of p66 and p51 were concentration-dependent with second-order rate constants (k(ass)) of approximately 1.7 M(-1) s(-1) for the simple bimolecular association reaction. The implications of slow dimerization of p66/p51 for the maturation process are discussed. A reaction-controlled model invoking conformational selection is proposed to explain the slow protein-protein association kinetics.

Dependence of tryptophan emission wavelength on conformation in cyclic hexapeptides.

The wavelength of maximum emission of tryptophan depends on the local electrostatic environment of the indole chromophore. The time-resolved emission spectra of seven rigid cyclic hexapeptides containing a single tryptophan residue were measured. The emission maxima of the three decay-associated spectra for the seven peptides ranged from 341 to 359 nm, suggesting that different tryptophan rotamers have different emission maxima even in the case of solvent-exposed tryptophans. This conclusion is supported by quantum mechanical/molecular dynamics simulations of the six canonical side chain rotamers of tryptophan in solvated hexapeptides. The calculated range of emission maxima for the tryptophan rotamers of the seven peptides is 344-365 nm. The precision of the wavelength calculations and the peptide, water, and charged side chain contributions to the spectral shifts are examined. The results indicate that the emission maxima of decay-associated spectra can aid in the assignment of fluorescence lifetimes to tryptophan rotamers.

Endogenous tryptophan residues of cAPK regulatory subunit type IIbeta reveal local variations in environments and dynamics.

The amino terminal dimerization/docking domain and the two-tandem, carboxy-terminal cAMP-binding domains (A and B) of cAMP-dependent protein kinase regulatory (R) subunits are connected by a variable linker region. In addition to providing a docking site for the catalytic subunit, the linker region is a major source of sequence diversity between the R-subunit isoforms. The RIIbeta isoform uniquely contains two endogenous tryptophan residues, one at position 58 in the linker region and the other at position 243 in cAMP-binding domain A, which can act as intrinsic reporter groups of their dynamics and microenvironment. Two single-point mutations, W58F and W243F, allowed the local environment of each Trp to be probed using steady-state and time-resolved fluorescence techniques. We report that: (a) the tryptophan fluorescence of the wild-type protein largely reflects Trp243 emission; (2) cAMP selectively quenches Trp243 and thus acts as a cAMP sensor; (3) Trp58 resides in a highly solvated, unstructured, and mobile region of the protein; and (4) Trp243 resides in a stable, folded domain and is relatively buried and rigid within the domain. The use of endogenous Trp residues presents a non-perturbing method for studying R-subunit subdomain characteristics in addition to providing the first biophysical data on the RIIbeta linker region.

Correlation of tryptophan fluorescence spectral shifts and lifetimes arising directly from heterogeneous environment.

Tryptophan (Trp) fluorescence is potentially a powerful probe for studying the conformational ensembles of proteins in solution, as it is highly sensitive to the local electrostatic environment of the indole side chain. However, interpretation of the wavelength-dependent complex fluorescence decays of proteins has been stymied by controversy about two plausible origins of the typical multiple fluorescence lifetimes: multiple ground-state populations or excited-state relaxation. The latter naturally predicts the commonly observed wavelength-lifetime correlation between decay components, which associates short lifetimes with blue-shifted emission spectra and long lifetimes with red-shifted spectra. Here we show how multiple conformational populations also lead to the same strong wavelength-lifetime correlation in cyclic hexapeptides containing a single Trp residue. Fluorescence quenching in these peptides is due to electron transfer. Quantum mechanics-molecular mechanics simulations with 150-ps trajectories were used to calculate fluorescence wavelengths and lifetimes for the six canonical rotamers of seven hexapeptides in aqueous solution at room temperature. The simulations capture most of the unexpected diversity of the fluorescence properties of the seven peptides and reveal that rotamers having blue-shifted emission spectra, i.e., higher average energy, have an increased probability for quenching, i.e., shorter average lifetime, during large fluctuations in environment that bring the nonfluorescent charge transfer state and the fluorescing state into resonance. This general mechanism should also be operative in proteins that exhibit multiexponential fluorescence decays, where myriad other sources of conformational heterogeneity besides rotamers are possible.

Effects of efavirenz binding on the subunit equilibria of HIV-1 reverse transcriptase.

Recent studies showed that nonnucleoside reverse transcriptase inhibitors (NNRTIs) have variable effects on dimerization of p66 and p51 subunits of HIV-1 reverse transcriptase (RT). Efavirenz, one of three NNRTIs currently used in highly active anti-retroviral therapy, enhances subunit dimerization. Sedimentation equilibrium experiments on each subunit and equimolar mixtures of both subunits were used to measure dissociation constants for the three coupled dimerization reactions of RT in the absence and presence of saturating concentrations of the drug. The dimerization constants of the p51/p51 homodimer, the p66/p66 homodimer, and the p66/p51 heterodimer increased 600-, 50-, and 25-fold, respectively, upon binding of efavirenz. The effects of NNRTIs on RT dimerization are consistent with a thermodynamic linkage between subunit association/dissociation and inhibitor binding. Analysis of crystal structures of the p66/p51 heterodimer reveals that efavirenz binding induces small structural changes at the dimer interface.

Attenuation of DNA replication by HIV-1 reverse transcriptase near the central termination sequence.

Previous pre-steady-state kinetic studies of equine infectious anemia virus-1 (EIAV) reverse transcriptase (RT) showed two effects of DNA substrates containing the central termination sequence (CTS) on the polymerization reaction: reduction of burst amplitude in single nucleotide addition experiments and accumulation of termination products during processive DNA synthesis [Berdis, A. J., Stetor, S. R., Le Grice, S. F. J., and Barkley, M. D. (2001) Biochemistry 40, 12140-12149]. The present study of HIV RT uses pre-steady-state kinetic techniques to evaluate the molecular mechanisms of the lower burst amplitudes using both random sequence and CTS-containing DNA substrates. The effects of various factors, including primer/template length, binding orientation, and protein concentration, on the burst amplitude were determined using random sequence DNA substrates. The percent active RT increases with total RT concentration, indicating that reversible dissociation of RT dimer is responsible for substoichiometric burst amplitudes with normal substrates. This finding was confirmed by gel mobility shift assays. Like EIAV RT, HIV RT showed lower burst amplitudes on CTS-containing DNA substrates compared to random sequences. The dissociation kinetics of RT-DNA complexes were monitored by enzyme activity and fluorescence. Biphasic kinetics were observed for both random sequence and CTS-containing DNA complexes, revealing two forms of the RT-DNA complex. A mechanism is proposed to account for reduction in burst amplitude of CTS-containing DNA that is consistent with the results of both single nucleotide addition and dissociation experiments. The two forms of the RT-DNA complex may represent partitioning of primer/template between the P- and N-sites on RT for the nucleic acid substrate.

Conformational effects on tryptophan fluorescence in cyclic hexapeptides.

The peptide bond quenches tryptophan fluorescence by excited-state electron transfer, which probably accounts for most of the variation in fluorescence intensity of peptides and proteins. A series of seven peptides was designed with a single tryptophan, identical amino acid composition, and peptide bond as the only known quenching group. The solution structure and side-chain chi(1) rotamer populations of the peptides were determined by one-dimensional and two-dimensional (1)H-NMR. All peptides have a single backbone conformation. The -, psi-angles and chi(1) rotamer populations of tryptophan vary with position in the sequence. The peptides have fluorescence emission maxima of 350-355 nm, quantum yields of 0.04-0.24, and triple exponential fluorescence decays with lifetimes of 4.4-6.6, 1.4-3.2, and 0.2-1.0 ns at 5 degrees C. Lifetimes were correlated with ground-state conformers in six peptides by assigning the major lifetime component to the major NMR-determined chi(1) rotamer. In five peptides the chi(1) = -60 degrees rotamer of tryptophan has lifetimes of 2.7-5.5 ns, depending on local backbone conformation. In one peptide the chi(1) = 180 degrees rotamer has a 0.5-ns lifetime. This series of small peptides vividly demonstrates the dominant role of peptide bond quenching in tryptophan fluorescence.

Tailoring nutrition education intervention programs to meet needs and interests of older adults.

Methods for determining appropriate content of older adult nutrition education intervention programs and strategies for effectively delivering nutrition messages to older learners are presented. Educators can determine the nutrition education needs and interests of their older learners by using results of food intake surveys and assessment screening tools, written surveys, interviews and group discussions. Findings of recent reports using these methods are summarized. Additionally, published experiences with and suggestions for tailoring education intervention programs for older adult audiences, including those of particular racial/ethnic groups, are reviewed. The need for research in this area is presented. This article is one of a series of literature reviews of topics related to nutrition education for older adults.

Concepts, theories and design components for nutrition education programs aimed at older adults.

This article examines characteristics of older adult learners and discusses adult education theory and empowerment concepts, along with nutrition education and behavioral change strategies for older adult nutrition education programs. Design components for older adult nutrition education programs are presented. Educational and behavioral change strategies should be selected based on characteristics of the intended audience, including their nutrition needs, wants and desires, and should be based on appropriate theory. Multi-disciplinary research is needed to develop behavioral and educational theoretical frameworks, as well as designs, intervention strategies, and evaluation methods for educational programs that lead to older adults adopting more healthful nutrition practices. This is one of a series of recent literature reviews on nutrition education for older adults.

Improving effectiveness of nutrition education resources for older adults.

This article discusses published reports and studies from the past decade that focused primarily on using written or other tangible nutrition educational resources with older adults, such as brief "handouts," newsletters, brochures, booklets, curricular lessons, board games, audiotapes and videotapes. Studies of health professionals' needs and desires for such materials are also reviewed. Thirteen articles, of which four were theory-based, were found. They are summarized in tables that include details regarding the educational resource(s) used; a description of the subjects and controls, if any; the evaluation methods used; and results obtained. Ten practical suggestions are offered to help educators select or develop more effective printed, audiotape, videotape and other tangible nutrition education resources appropriate for older adults. Much research remains to be done in this area. This article is one of a series of literature reviews of topics related to nutrition education for older adults.

Group nutrition education classes for older adults.

A thorough search of the literature revealed only nine articles published since 1993 that focused on nutrition education for older adults attending group classes and that measured outcomes. A table summarizes the reports, including the theoretical bases, descriptions of interventions, participants and comparison groups, program outcomes, methods of verification, and follow-up after interventions. Only three of the studies explicitly indicated that elements of a stated behavioral change theory had been incorporated. All of the educators employed a variety of older adult educational strategies to enhance learning. Six research teams reported on classes where nearly half or more of the participants represented minority groups. Six studies included comparison groups. Types of outcomes included measurements of change in knowledge, attitudes/ beliefs, behaviors, and/or physiological measures, but the actual variables examined differed among reports. No consistent patterns were detected among reported outcomes. The longest follow-up after interventions ceased was seven months. The review addresses issues raised from an analysis of the quantity, quality and findings of the articles and makes suggestions for future research and offers preliminary ideas for developing group nutrition education classes for older adults. This is one of a series of reviews of recent literature on nutrition education for older adults.

Barriers to nutrition education for older adults, and nutrition and aging training opportunities for educators, healthcare providers,volunteers and caregivers.

Literature citations of barriers to nutrition education found in those who teach and care for older adults, as well as within older adults themselves, are discussed. No attempt was made to compare educational barriers for learners of varying ages. These obstacles need to be addressed in order for nutrition to be taught or learned effectively so that nutrition practices and health improve. Barriers for healthcare professionals to providing nutrition education include misconceptions and stereotypes about older adults and about their nutritional concerns; lack of attention to and lack of funding for older adult educational programs; and difficulties recruiting older learners. Hindrances for older adults in responding to nutrition education can be categorized as attitudinal, motivational, environmental, and related to low literacy and poverty. Published examples of opportunities for education and training about nutrition and aging that are in place for health educators, healthcare providers, volunteers and caregivers regarding nutrition and aging are discussed. Suggestions are presented regarding future efforts to minimize educational barriers and to provide training for healthcare professionals, volunteers and caregivers. New research is needed in this field of study in order to realize the potential quality of life benefits and reduced healthcare costs associated with providing effective nutrition education to older adults. This is one of a series of reviews of recent literature on nutrition education for older adults.

Two-step FRET as a structural tool.

The power of FRET to study molecular complexes is expanded by the use of two or more donor/acceptor pairs. A general theoretical framework for distance measurements in three-chromophore systems is presented. Three energy transfer schemes applicable to many diverse situations are considered: (I) two-step FRET relay with FRET between the first and second chromophores and between the second and third, (II) FRET from a single donor to two different acceptors, and (III) two-step FRET relay with FRET also between the first and third chromophores. Equations for the efficiencies involving multiple energy transfer steps are derived for both donor quenching and sensitized emission measurements. The theory is supported by experimental data on model systems of known structure using steady-state donor quenching, lifetime quenching, and sensitized emission. The distances measured in the three-chromophore systems agree with those in two-chromophore systems and molecular models. Finally, labeling requirements for diagnosis of the energy transfer scheme and subsequent distance measurements are discussed.