In Vivo Activity of Repurposed Amodiaquine as a Host-Targeting Therapy for the Treatment of Anthrax.

Anthrax is caused by Bacillus anthracis and can result in nearly 100% mortality due in part to anthrax toxin. Antimalarial amodiaquine (AQ) acts as a host-oriented inhibitor of anthrax toxin endocytosis. Here, we determined the pharmacokinetics and safety of AQ in mice, rabbits, and humans as well as the efficacy in the fly, mouse, and rabbit models of anthrax infection. In the therapeutic-intervention studies, AQ nearly doubled the survival of mice infected subcutaneously with a B. anthracis dose lethal to 60% of the animals (LD60). In rabbits challenged with 200 LD50 of aerosolized B. anthracis, AQ as a monotherapy delayed death, doubled the survival rate of infected animals that received a suboptimal amount of antibacterial levofloxacin, and reduced bacteremia and toxemia in tissues. Surprisingly, the anthrax efficacy of AQ relies on an additional host macrophage-directed antibacterial mechanism, which was validated in the toxin-independent Drosophila model of Bacillus infection. Lastly, a systematic literature review of the safety and pharmacokinetics of AQ in humans from over 2 000 published articles revealed that AQ is likely safe when taken as prescribed, and its pharmacokinetics predicts anthrax efficacy in humans. Our results support the future examination of AQ as adjunctive therapy for the prophylactic anthrax treatment.

Microimplant-assisted rapid palatal expansion appliance to orthopedically correct transverse maxillary deficiency in an adult.

This case report describes the use of a microimplant-assisted rapid palatal expansion (MARPE) appliance to orthopedically correct a transverse maxillary deficiency in an adult patient. Expansion forces transmitted through the teeth in traditional rapid palatal expansion appliances create unwanted dental effects rather than true skeletal expansion, particularly in older patients with more rigid interdigitation of the midpalatal suture. This 19-year-old patient had maxillary constriction with a unilateral posterior crossbite. A MARPE appliance secured to the palatal bones with 4 microimplants was expanded by 10 mm. Pre-MARPE and post-MARPE cone-beam computed tomography cross sections demonstrated 4 to 6 mm of expansion of the maxillofacial structures, including the zygoma and nasal bone area, and widening of the circummaxillary sutures. Minor buccal tipping of the dentition was observed, but the integrity of the alveolar bone was preserved. This report demonstrates that careful design and application of the MARPE appliance can achieve successful transverse expansion of the maxilla and the surrounding structures in a patient beyond the age typically considered acceptable for traditional rapid palatal expansion.

Neutralizing antibody and functional mapping of Bacillus anthracis protective antigen-The first step toward a rationally designed anthrax vaccine.

Anthrax is defined by the Centers for Disease Control and Prevention as a Category A pathogen for its potential use as a bioweapon. Current prevention treatments include Anthrax Vaccine Adsorbed (AVA). AVA is an undefined formulation of Bacillus anthracis culture supernatant adsorbed to aluminum hydroxide. It has an onerous vaccination schedule, is slow and cumbersome to produce and is slightly reactogenic. Next-generation vaccines are focused on producing recombinant forms of anthrax toxin in a well-defined formulation but these vaccines have been shown to lose potency as they are stored. In addition, studies have shown that a proportion of the antibody response against these vaccines is focused on non-functional, non-neutralizing regions of the anthrax toxin while some essential functional regions are shielded from eliciting an antibody response. Rational vaccinology is a developing field that focuses on designing vaccine antigens based on structural information provided by neutralizing antibody epitope mapping, crystal structure analysis, and functional mapping through amino acid mutations. This information provides an opportunity to design antigens that target only functionally important and conserved regions of a pathogen in order to make a more optimal vaccine product. This review provides an overview of the literature related to functional and neutralizing antibody epitope mapping of the Protective Antigen (PA) component of anthrax toxin.

Presentation of peptides from Bacillus anthracis protective antigen on Tobacco Mosaic Virus as an epitope targeted anthrax vaccine.

The current anthrax vaccine requires improvements for rapidly invoking longer-lasting neutralizing antibody responses with fewer doses from a well-defined formulation. Designing antigens that target neutralizing antibody epitopes of anthrax protective antigen, a component of anthrax toxin, may offer a solution for achieving a vaccine that can induce strong and long lasting antibody responses with fewer boosters. Here we report implementation of a strategy for developing epitope focused virus nanoparticle vaccines against anthrax by using immunogenic virus particles to present peptides derived from anthrax toxin previously identified in (1) neutralizing antibody epitope mapping studies, (2) toxin crystal structure analyses to identify functional regions, and (3) toxin mutational analyses. We successfully expressed two of three peptide epitopes from anthrax toxin that, in previous reports, bound antibodies that were partially neutralizing against toxin activity, discovered cross-reactivity between vaccine constructs and toxin specific antibodies raised in goats against native toxin and showed that antibodies induced by our vaccine constructs also cross-react with native toxin. While protection against intoxication in cellular and animal studies were not as effective as in previous studies, partial toxin neutralization was observed in animals, demonstrating the feasibility of using plant-virus nanoparticles as a platform for epitope defined anthrax vaccines.

Identification of agents effective against multiple toxins and viruses by host-oriented cell targeting.

A longstanding and still-increasing threat to the effective treatment of infectious diseases is resistance to antimicrobial countermeasures. Potentially, the targeting of host proteins and pathways essential for the detrimental effects of pathogens offers an approach that may discover broad-spectrum anti-pathogen countermeasures and circumvent the effects of pathogen mutations leading to resistance. Here we report implementation of a strategy for discovering broad-spectrum host-oriented therapies against multiple pathogenic agents by multiplex screening of drugs for protection against the detrimental effects of multiple pathogens, identification of host cell pathways inhibited by the drug, and screening for effects of the agent on other pathogens exploiting the same pathway. We show that a clinically used antimalarial drug, Amodiaquine, discovered by this strategy, protects host cells against infection by multiple toxins and viruses by inhibiting host cathepsin B. Our results reveal the practicality of discovering broadly acting anti-pathogen countermeasures that target host proteins exploited by pathogens.

Early mixed dentition treatment of bilateral impaction of permanent mandibular first molars.

Available scientific literature is scarce in covering cases involving impaction of permanent mandibular first molars treated in mixed dentition patients. The purpose of this report was to present the case of a 7-year-old patient with bilateral impaction of permanent mandibular first molars treated early using a simple and effective removable appliance. The proposed intervention involved the construction of a removable acrylic appliance with bilateral titanium-molybdenum alloy distalizing springs to disimpact and allow complete eruption of the molars. A button was bonded to the occlusal surface of each molar to be used as a leverage point for the distalizing springs. Following seven months of treatment, both the permanent mandibular right and left first molars were upright and in physiologic position. This case demonstrates that, when this problem is identified and treated early, pediatric dentists have the opportunity to minimize the complexity of future orthodontic treatment and limit the extent of malocclusion in the permanent dentition.

Influence of the vertical position of maxillary central incisors on the perception of smile esthetics among orthodontists and laypersons.

OBJECTIVE: The purpose of this study was to determine the perception of smile esthetics among orthodontists and laypeople with respect to different upper central incisor vertical positions in a frontal smile analysis. MATERIALS AND METHODS: A frontal close-up smile photo of an adult Caucasian woman was selected. The patient had healthy upper anterior dentition and had no history of orthodontic treatment. Images were altered in order to create six different central incisor vertical positions in 0.5-mm increments. All images were assessed in three different views: full smile, gingival close-up excluding incisal edges, and incisal close-up excluding gingival margins. Images were randomly assembled in an album, which was given to 120 judges: 60 orthodontists and 60 laypersons. Each rater was asked to evaluate the attractiveness of the images using the visual analog scale. The data collected were then statistically analyzed. RESULTS: The highest rated smiles showed two notable characteristics: the central-to-lateral incisal step was 1.5 mm; and the central incisor gingival margins matched the laterals, and both were 0.5 mm below the line of the canine gingival margins. The least attractive smile was the one with no step between the centrals and laterals, and with the central incisor gingival margins 1.0 mm above the canine gingival margins. CONCLUSION: The results of this study indicate that slightly extruded upper central incisors are more esthetically preferred than intruded. CLINICAL SIGNIFICANCE: The upper central incisors are the key determinant in evaluating smile esthetics, and thus, the assessment of their ideal vertical positioning is an aspect of paramount importance.

Predictors of velopharyngeal insufficiency after Le Fort I maxillary advancement in patients with cleft palate.

PURPOSE: Approximately 25% to 40% of patients with cleft lip/palate develop maxillary retrusion that requires Le Fort I osteotomy. Maxillary advancement brings the soft palate forward, and this may cause velopharyngeal insufficiency (VPI). The goal of this study was to identify predictors that place patients with repaired cleft palate at risk of developing VPI after Le Fort I advancement. MATERIALS AND METHODS: This was a retrospective study of nonsyndromic patients with cleft lip/palate who had a Le Fort I osteotomy between 2000 and 2008. Charts were reviewed and data were collected on patient characteristics, preoperative speech assessments, and nasopharyngoscopic reports. Pre- and postoperative cephalometric radiographs were used to measure maxillary advancement and to assess the structure of the velopharynx. Simple logistic regression analysis examined the association between each predictive variable and postoperative VPI, as indicated by need for pharyngeal flap. Predictors with P ≤ .10 were included in the multivariate regression model. In both the univariate and the multivariate analyses, P ≤ .05 was considered statistically significant. RESULTS: Univariate analysis showed a significant association between preoperative soft palatal length and need for a pharyngeal flap (P = .005). By multivariate analysis, both preoperative soft palatal length and postoperative pharyngeal depth were associated with need for pharyngeal flap (P = .003 and P = .030). CONCLUSION: This study shows that a short soft palate is associated with VPI after Le Fort I osteotomy. Assessment of palatal length and pharyngeal depth on cephalometric radiographs is helpful in predicting postoperative VPI and need for a pharyngeal flap in patients with cleft palate after maxillary advancement.