Etiologies of Acute Undifferentiated Febrile Illnesses in and near Iquitos from 1993 to 1999 in the Amazon River Basin of Peru.

The objective of this study was to determine the etiology of febrile illnesses among patients from October 1, 1993 through September 30, 1999, in the urban community of Iquitos in the Amazon River Basin of Peru. Epidemiological and clinical data as well as blood samples were obtained from consenting patients at hospitals, health clinics and private residences. Samples were tested for arboviruses in cell cultures and for IgM and IgG antibodies by ELISA. Blood smears were examined for malaria, and sera were tested for antibodies to Leptospira spp. by ELISA and microscopic agglutination. Among 6,607 febrile patients studied, dengue viruses caused 14.6% of the cases, and Venezuelan equine encephalitis virus caused 2.5%, Oropouche virus 1.0%, Mayaro virus 0.4%, and other arboviruses caused 0.2% of the cases. Also, 22.9% of 4,844 patients tested were positive for malaria, and of 400 samples tested, 9% had evidence of acute leptospirosis. Although the study was not designed to assess the importance of these pathogens as a cause of human morbidity in the total population, these results indicate that arboviruses, leptospirosis, and malaria were the cause of approximately 50% of the febrile cases. Although the arboviruses that were diagnosed can produce asymptomatic infections, our findings increased the overall understanding of the relative health burden of these infections, as well as baseline knowledge needed for designing and implementing further studies to better assess the health impact and threat of these pathogens in the Amazon Basin of Peru.

Reliability of meniscus tear description: a study using MRI from the Osteoarthritis Initiative.

Despite being one of the most common indications for surgery, data on the types of meniscus tear that should be treated surgically are limited. Improving patient selection requires agreement on meniscus tear description. This study evaluates a simple MRI tear classification system for inter-observer agreement. Knee MRI examinations from 57 subjects from the Osteoarthritis Initiative cohort were reviewed by two sub-specialty trained, musculoskeletal radiologists. Based on two pulse sequences, each meniscus was classified by: tear or no tear; location of tear in anterior, middle or posterior third or multiple thirds; and displaced or non-displaced radial, horizontal, longitudinal or complex tear pattern. A tear was defined as signal abnormality extending to the surface on at least two images and displacement as more than 2 mm of extrusion or separation measured orthogonal to the tear plane. Kappa, weighted Kappa and percentage agreement were calculated. For the medial meniscus, Kappa and percentage agreement estimates were, respectively: the presence of tear, 0.79 and 89.5%; tear with displacement, 0.70 (weighted Kappa) and 66.0%; tear description, 0.47 and 61.4%; tear location, 0.64 and 79.0%. For the lateral meniscus, estimates were: the presence of tear, 0.75 and 89.5%; tear with displacement, 0.81 (weighted Kappa) and 86.0%; tear description, 0.56 and 78.9%; tear location, 0.74 and 87.7%. The strength of agreement between readers was moderate to substantial underscoring the challenge of meniscus tear classification.

Correlation between the accessory anterolateral talar facet, bone marrow edema, and tarsal coalitions.

OBJECTIVE: The purposes of this study was to determine the prevalence of the accessory anterolateral talar facet (AALTF); to evaluate the relationship between AALTF, focal abutting bone marrow edema (FABME), and sinus tarsi edema; and to study the prevalence of tarsal coalitions in patients with the AALTF utilizing ankle MR images. MATERIALS AND METHODS: 5-T ankle MR images were reviewed for the presence of AALTF, FABME, sinus tarsi edema, tarsal coalition, and location and type of coalition (cartilaginous, fibrous, and osseous). Multivariate analysis was performed to examine the correlation between AALTF and the other variables. RESULTS: Three hundred ninety-one consecutive patients were included (age range 5-86 years; mean age 45 years). An AALTF was present in 3.6% (14/391) of patients. The AALTF prevalence was 2% in women and 6.6% in men. There was a significantly association between AALTF and FABME (9/14, p < 0.05), sinus tarsi edema (13/14, p < 0.05), and tarsal coalition (4/14, p < 0.05). CONCLUSION: AALTF is relatively often detected on MRI of the ankle and significantly associated with BME, sinus tarsi edema, and subtalar coalition. Patients with a tarsal coalition should be evaluated for the concurrent presence of an AALTF.

Case report: vertebral foreign body granuloma mimicking a skeletal metastasis.

Intraosseous foreign body granuloma formation related to migrated surgical material is a rarely reported condition with variable imaging appearance. In this case report, we describe a foreign body granuloma that occurred in a lumbar vertebral body one level above a prior surgical fusion. The lytic appearance mimicked a skeletal metastasis in a 65-year-old patient with recently diagnosed renal cell carcinoma. To the best of our knowledge, this is the first reported case of a lumbar vertebral foreign body granuloma occurring distant from the site of surgery, indistinguishable from skeletal metastasis on radiologic examination.

A novel approach for determining three-dimensional acetabular orientation: results from two hundred subjects.

BACKGROUND: The inherently complex three-dimensional morphology of both the pelvis and acetabulum create difficulties in accurately determining acetabular orientation. Our objectives were to develop a reliable and accurate methodology for determining three-dimensional acetabular orientation and to utilize it to describe relevant characteristics of a large population of subjects without apparent hip pathology. METHODS: High-resolution computed tomography studies of 200 patients previously receiving pelvic scans for indications not related to orthopaedic conditions were selected from our institution's database. Three-dimensional models of each osseous pelvis were generated to extract specific anatomical data sets. A novel computational method was developed to determine standard measures of three-dimensional acetabular orientation within an automatically identified anterior pelvic plane reference frame. Automatically selected points on the osseous ridge of the acetabulum were used to generate a best-fit plane for describing acetabular orientation. RESULTS: Our method showed excellent interobserver and intraobserver agreement (an intraclass correlation coefficient [ICC] of >0.999) and achieved high levels of accuracy. A significant difference between males and females in both anteversion (average, 3.5°; 95% confidence interval [CI], 1.9° to 5.1° across all angular definitions; p < 0.0001) and inclination (1.4°; 95% CI, 0.6° to 2.3° for anatomic angular definition; p < 0.002) was observed. Intrapatient asymmetry in anatomic measures showed bilateral differences in anteversion (maximum, 12.1°) and in inclination (maximum, 10.9°). CONCLUSIONS: Significant differences in acetabular orientation between the sexes can be detected only with accurate measurements that account for the entire acetabulum. While a wide range of interpatient acetabular orientations was observed, the majority of subjects had acetabula that were relatively symmetrical in both inclination and anteversion. CLINICAL RELEVANCE: A highly accurate and reproducible method for determining the orientation of the acetabulum's aperture will benefit both surgeons and patients, by further refining the distinctions between normal and abnormal hip characteristics. Enhanced understanding of the acetabulum could be useful in the diagnostic, planning, and execution stages for surgical procedures of the hip or in advancing the design of new implant systems.

Use of a validated algorithm to judge the appropriateness of total knee arthroplasty in the United States: a multicenter longitudinal cohort study.

OBJECTIVE: In previous studies conducted outside the US, ∼20% of total knee arthroplasty (TKA) surgeries were judged to be inappropriate. The present study was undertaken to determine the prevalence rates of TKA surgeries classified as appropriate, inconclusive, and inappropriate in a knee osteoarthritis population in the US. METHODS: We used a modification of a validated appropriateness classification system and applied it to patients in the Osteoarthritis Initiative data set who underwent TKA. A variety of preoperative data were used in the classification, including Western Ontario and McMaster Universities Osteoarthritis Index pain and physical function scores, radiographic features, knee motion and laxity measures, and age. RESULTS: Data on 205 patients who underwent TKA were examined. The prevalence rates for classification of the procedure as appropriate, inconclusive, and inappropriate were 44.0% (95% confidence interval [95% CI] 37-51%), 21.7% (95% CI 16-28%), and 34.3% (95% CI 27-41%), respectively. CONCLUSION: Approximately one-third of TKA surgeries were judged to be inappropriate. Variation in the characteristics of patients undergoing TKA was extensive. These data support the need for consensus development of criteria for patient selection among US practitioners treating patients who are potential candidates for TKA. Among the important issues, consensus development needs to address variation in patient characteristics and the relative importance of preoperative status and subsequent outcome.

Plantar measurements to determine success of surgical correction of Stage IIb adult acquired flatfoot deformity.

Adult acquired flatfoot deformity is a degenerative disease causing medial arch dysfunction. Surgical correction has typically involved tendon reconstruction with calcaneal osteotomy; however, the postoperative changes have not been fully characterized. The present study assessed the success of surgical correction of Stage IIb adult acquired flatfoot deformity through changes in plantar pressures and patient-generated outcome scores. With Institutional Review Board approval, 6 participants were evaluated before and after surgery using pedobarography, the Foot and Ankle Outcome Score, and the Medical Outcomes Study 36-item short-form questionnaire. The plantar pressures were recorded using a TekScan HRMat(®) during walking and in a 1- and 2-foot stance. The resulting contour maps were segmented into 9 regions, with the peak pressure, normalized force, and arch index calculated. Surgical effects were analyzed using paired t tests. Postoperatively, the Foot and Ankle Outcome Score and Medical Outcomes Study 36-item short-form questionnaire scores increased significantly from 180 ± 78 to 360 ± 136 (p < .03) and 47 ± 18 to 71 ± 19 (p = .06), respectively. During the 2-foot stance, the normalized force had increased significantly in the lateral midfoot (p < .03), although no significant differences were found in peak pressures. No significant differences were observed in the 1-foot stance. During walking, the normalized force increased significantly in the lateral mid- and forefoot (p < .05). The peak pressure increased significantly in the lateral forefoot (p < .01). The arch index values demonstrated no significant changes. The increased questionnaire scores indicated that surgical correction improved the self-perceived health of the participants. Lateral shifts in the peak pressure and normalized force suggest that forefoot and midfoot loading is altered postoperatively, consistent with the goal of offloading the dysfunctional arch. Thus, the present study has demonstrated that surgical treatment of adult acquired flatfoot deformity can be accurately assessed using patient-reported outcome measures and plantar pressures.

Validation of a population of patient-specific adult acquired flatfoot deformity models.

Adult acquired flatfoot deformity (AAFD) is a degenerative disease resulting in malalignment of the mid- and hindfoot secondary to posterior tibial tendon dysfunction and increasing implication of ligament pathologies. Despite the complex 3D nature of AAFD, 2D radiographs are still employed to diagnose and stage the disease. Computer modeling techniques allow for accurate 3D recreations of musculoskeletal systems for the investigation of biomechanical factors contributing to disease. Following Institutional Review Board approval, the lower limbs of six diagnosed AAFD sufferers were imaged with MRI, photographs, and X-ray. Next, a radiologist graded the MRI attenuation of eight soft-tissues implicated in AAFD. Six patient-specific rigid-body models were then created and loaded according to patient weight, graded soft-tissues, and extrinsic muscles. Model function was validated using clinically relevant kinematic measures in three planes. Agreement varied depending on the measure, with average absolute deviations of < 7° for angles and <4 mm for distances. Additionally, the clinically favored AP talonavicular coverage angle, ML talo-1st metatarsal angle, and ML 1st cuneiform height showed strong correlations of R(2) = 0.63, 0.75, and 0.85, respectively. Thus, computer modeling offers a promising methodology for the non-invasive investigation of in vivo kinematic behavior in pathologic feet and, once validated, may further be used to investigate biomechanical parameters that are difficult to measure clinically.

Imaging of glenoid labrum lesions.

This article reviews the current status of the imaging of the glenoid labrum and associated structures, including anatomic variants and the different types of labral disease.

Considerations when designing a disease-modifying osteoarthritis drug (DMOAD) trial using radiography.

OBJECTIVE: Using placebo data from a recently completed disease-modifying osteoarthritis (OA) drug trial, we seek to inform study design of future radiographic studies. METHODS: Eligible patients aged ≥40 years, with body mass index (BMI) 25-40kg/m(2) and symptomatic knee OA diagnosed by modified Kellgren and Lawrence grade (KLG) 2 or 3 and pain/stiffness and/or use of medication for knee pain in the past year, were assessed by radiography using a modified Lyon-schuss (mL/S) protocol for joint space narrowing (JSN) (primary outcome variable) at baseline and weeks 48 and 96. Multifaceted quality control was conducted throughout. Repeat images were requested when the medial tibial plateau (MTP) was not aligned (inter-margin distance [IMD] >1.5mm) or for other quality issues. Data are given mean ± standard deviation. RESULTS: Patients (74.9% female; 61.3 ± 9.1 years) had BMI 31.6 ± 4.1kg/m(2) at baseline; 222 (173 females) had KLG2, 264 (191 female) KLG3. A significant loss in joint space width (JSW) from baseline to week 48 (-0.13 ± 0.36mm) and to week 96 (-0.22 ± 0.45mm) was observed for all randomised placebo patients (p < 0.001 for both), and at both time points when stratified by KLG2 or KLG3. Standard deviations were small relative to mean changes, providing standardised response means for all placebo patients of 0.35 (week 48) and 0.48 (week 96). CONCLUSIONS: Using a tightly controlled radiographic technique, JSN is a viable outcome variable for determining disease progression in mild-to-moderate knee OA. The mL/S protocol is a sensitive and feasible method for OA studies aiming to assess rate of JSN in the knee.

A 2-year randomised, double-blind, placebo-controlled, multicentre study of oral selective iNOS inhibitor, cindunistat (SD-6010), in patients with symptomatic osteoarthritis of the knee.

OBJECTIVE: To determine if inhibition of inducible nitric oxide synthase (iNOS) with cindunistat hydrochloride maleate slows progression of osteoarthritis (OA) METHODS: This 2-year, multinational, double-blind, placebo-controlled trial enrolled patients with symptomatic knee OA (Kellgren and Lawrence Grade (KLG) 2 or 3). Standard OA therapies were permitted throughout. Patients were randomly assigned to cindunistat (50 or 200 mg/day) or placebo. Randomisation was stratified by KLG. Radiographs to assess joint space narrowing (JSN) were acquired using the modified Lyon-schuss protocol at baseline, week 48 and 96. RESULTS: Of 1457 patients (50 mg/day, n=485; 200 mg/day, n=486; placebo, n=486), 1048 (71.9%) completed the study. Patients were predominantly women; 56% had KLG3. The primary analysis did not demonstrate superiority of cindunistat versus placebo for rate of change in JSN. In KLG2 patients, JSN after 48 weeks was lower with cindunistat 50 mg/day versus placebo (p=0.032). Least-squares mean±SE JSN with cindunistat 50 mg/day ( -0.048±0.028 mm) and 200 mg/day (-0.062±0.028 mm) were 59.9% (95% CI 6.8% to 106.9%) and 48.7% (95% CI -8.4% to 93.9%) of placebo, improvement was not maintained at 96 weeks. No improvement was observed for KLG3 patients at either time-point. Cindunistat did not improve joint pain or function, but was generally well tolerated. CONCLUSIONS: Cindunistat (50 or 200 mg/day) did not slow the rate of JSN versus placebo. After 48-weeks, KLG2 patients showed less JSN; however, the improvement was not sustained at 96-weeks. iNOS inhibition did not slow OA progression in KLG3 patients. CLINICAL TRIAL LISTING: NCT00565812.

ACR appropriateness criteria(®) acute trauma to the knee.

There are more than 1 million visits to the ER annually in the United States for acute knee trauma. Many of these are twisting injuries in young patients who can walk and bear weight, and emergent radiography is not required. Several clinical decision rules have been devised that can considerably reduce the number of radiographic studies ordered without missing a clinically significant fracture. Although fractures are seen on only 5% of emergency department knee radiographs, 86% of knee fractures result from blunt trauma. In patients with falls or twisting injuries who have focal tenderness, effusion, or inability to bear weight, radiography should be the first imaging study performed. If radiography shows no fracture, MRI is best for evaluating for a suspected meniscal or ligament tear or patellar dislocation. Patients with knee dislocation should undergo radiography and MRI, as well as fluoroscopic angiography, CT angiography, or MR angiography. The ACR Appropriateness Criteria(®) are evidence-based guidelines for specific clinical conditions that are reviewed every 2 years by a multidisciplinary expert panel. The guideline development and review include an extensive analysis of current medical literature from peer-reviewed journals and the application of a well-established consensus methodology (modified Delphi) to rate the appropriateness of imaging and treatment procedures by the panel. In those instances in which evidence is lacking or not definitive, expert opinion may be used to recommend imaging or treatment.

Immunogenicity and protective efficacy of a vaxfectin-adjuvanted tetravalent dengue DNA vaccine.

A prototype dengue-1 DNA vaccine was shown to be safe and immunogenic in a previous Phase 1 clinical trial. Anti-dengue-1 neutralizing antibody responses were detectable only in the group of volunteers receiving the high dose of nonadjuvanted vaccine and the antibody titers were low. Vaxfectin(®), a lipid-based adjuvant, enhances the immunogenicity of DNA vaccines. We conducted a nonhuman primate study to evaluate the effect of Vaxfectin(®) on the immunogenicity of a tetravalent dengue DNA vaccine. Animals were immunized on days 0, 28 and 84, with each immunization consisting of 3mg of Vaxfectin(®)-adjuvanted tetravalent dengue DNA vaccine. The use of Vaxfectin(®) resulted in a significant increase in anti-dengue neutralizing antibody responses against dengue-1, -3 and -4. There was little to no effect on T cell responses as measured by interferon gamma ELISPOT assay. Animals immunized with the Vaxfectin(®)-formulated tetravalent DNA vaccine showed significant protection against live dengue-2 virus challenge compared to control animals (0.75 mean days of viremia vs 3.3 days). Animals vaccinated with nonadjuvanted DNA had a mean 2.0 days of viremia. These results support further evaluation of the Vaxfectin(®)-adjuvanted tetravalent dengue DNA vaccine in a Phase 1 clinical trial.

Evaluation of a prototype dengue-1 DNA vaccine in a Phase 1 clinical trial.

Candidate dengue DNA vaccine constructs for each dengue serotype were developed by incorporating pre-membrane and envelope genes into a plasmid vector. A Phase 1 clinical trial was performed using the dengue virus serotype-1 (DENV-1) vaccine construct (D1ME(100)). The study was an open-label, dose-escalation, safety and immunogenicity trial involving 22 healthy flavivirus-naïve adults assigned to one of two groups. Each group received three intramuscular injections (0, 1, and 5 months) of either a high dose (5.0mg, n=12) or a low dose (1.0mg, n=10) DNA vaccine using the needle-free Biojector(®) 2000. The most commonly reported solicited signs and symptoms were local mild pain or tenderness (10/22, 45%), local mild swelling (6/22, 27%), muscle pain (6/22, 27%) and fatigue (6/22, 27%). Five subjects (41.6%) in the high dose group and none in the low dose group developed detectable anti-dengue neutralizing antibodies. T-cell IFN gamma responses were detected in 50% (4/8) and 83.3% (10/12) of subjects in the low and high dose groups, respectively. The safety profile of the DENV-1 DNA vaccine is acceptable at both doses administered in the study. These results demonstrate a favorable reactogenicity and safety profile of the first in human evaluation of a DENV-1 DNA vaccine.

A review of the surgical treatment options for the obstructive sleep apnea/hypopnea syndrome patient.

Obstructive sleep apnea/hypopnea syndrome (OSAHS) is a medical condition that has received significant attention within the medical community and mainstream media due to its potentially serious physiological consequences and relatively frequent occurrence within the general population. From the military perspective, the impact on individual readiness for deployment, and the potential degradation of performance in critically important military duties, often results in tremendous expenditures of training resources, time, and expertise to replace the military member with a suitable substitute or release of the individual from active duty. This article reviews common surgical techniques for clinical management of OSAHS patients in a presentation format for primary care and sleep medicine specialists, as well as surgeons interested in the philosophies of surgical management of sleep disordered breathing. Presentation of risks and benefits of surgical treatment are discussed in a manner to facilitate communication between patient and health care provider.

Development of a clinical workflow tool to enhance the detection of vertebral fractures: accuracy and precision evaluation.

STUDY DESIGN: Image analysis model development. OBJECTIVE: The objective of this study was to develop a novel clinical workflow tool that uses model-based shape recognition technology to allow efficient, semiautomated detailed annotation of each vertebra between T4 and L4 on plain lateral radiographs. SUMMARY OF BACKGROUND DATA: Identification of prevalent vertebral fractures, especially when not symptomatic, has been problematic despite their importance. There is a recognized need to increase the opportunities to detect vertebral fractures so that clinically beneficial therapeutic interventions can be initiated. METHODS: Radiographs obtained from 165 subjects in the Canadian Multicenter Osteoporosis Study (CaMos) were used to construct a vertebral shape model of the vertebral column from T4 to L4 using a statistical learning technique, as well as to estimate the accuracy and precision of this automated software tool for vertebral shape analysis. Radiographs showing scoliosis greater than 15 degrees were excluded. RESULTS: Vertebral contours defined by 95 points per vertebra, represented by 79,895 points in total, were assessed on 841 individual vertebrae. The mean absolute accuracy error calculated over each vertebra in each test image was 1.06 +/- 1.2 mm. This value corresponded to an average 3.4% of vertebral height. The mean precision error, reflecting interobserver variability, per vertebra of the resulting annotations was 0.61 +/- 0.73 mm. This value corresponded to an average 2.3% of vertebral height. Accuracy and precision error estimates did not differ notably by vertebral level. CONCLUSION: The results of the current study indicate that statistical modeling can provide a robust tool for the accurate and precise semiautomated annotation of vertebral body shape from T4 to L4 in patients who do not have scoliosis greater than 15 degrees . This method may prove useful as a clinical workflow tool to aid the physician in vertebral fracture assessment and might contribute to decision-making about pharmacologic treatment of osteoporosis.

A dry-format field-deployable quantitative reverse transcriptase-polymerase chain reaction assay for diagnosis of dengue infections.

We have systematically evaluated a dry-format, quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) assay developed by Tetracore Inc. for the Cepheid SmartCycler platform to facilitate rapid diagnosis of dengue virus infections. A panel of related flaviviruses was used to evaluate the clinical specificity of the assay, and it was found to be specific to dengue. Eighty-one clinical samples previously confirmed dengue positive by virus isolation, along with 25 dengue negative control specimens were used to validate this new diagnostic assay. Using these clinical samples, the assay exhibited 98.77% sensitivity and 100% specificity. Over 85% of the clinical specimen exhibited viral loads ranging from 10(3) to 10(7) plaque-forming units per milliliter (PFU/mL). In addition, this dry-format assay is stable at ambient temperatures and requires minimal technical expertise to perform in a small thermocycler platform. These characteristics make it a promising candidate for diagnosis of dengue in mobile laboratories in the field.

A tetravalent dengue vaccine based on a complex adenovirus vector provides significant protection in rhesus monkeys against all four serotypes of dengue virus.

Nearly a third of the human population is at risk of infection with the four serotypes of dengue viruses, and it is estimated that more than 100 million infections occur each year. A licensed vaccine for dengue viruses has become a global health priority. A major challenge to developing a dengue vaccine is the necessity to produce fairly uniform protective immune responses to all four dengue virus serotypes. We have developed two bivalent dengue virus vaccines, using a complex adenovirus vector, by incorporating the genes expressing premembrane (prM) and envelope (E) proteins of dengue virus types 1 and 2 (dengue-1 and -2, respectively) (CAdVax-Den12) or dengue-3 and -4 (CAdVax-Den34). Rhesus macaques were vaccinated by intramuscular inoculation of a tetravalent dengue vaccine formulated by combining the two bivalent vaccine constructs. Vaccinated animals produced high-titer antibodies that neutralized all four serotypes of dengue viruses in vitro. The ability of the vaccine to induce rapid, as well as sustained, protective immune responses was examined with two separate live-virus challenges administered at 4 and 24 weeks after the final vaccination. For both of these virus challenge studies, significant protection from viremia was demonstrated for all four dengue virus serotypes in vaccinated animals. Viremia from dengue-1 and dengue-3 challenges was completely blocked, whereas viremia from dengue-2 and dengue-4 was significantly reduced, as well as delayed, compared to that of control-vaccinated animals. These results demonstrate that the tetravalent dengue vaccine formulation provides significant protection in rhesus macaques against challenge with all four dengue virus serotypes.

A heterologous DNA prime-Venezuelan equine encephalitis virus replicon particle boost dengue vaccine regimen affords complete protection from virus challenge in cynomolgus macaques.

A candidate vaccine (D1ME-VRP) expressing dengue virus type 1 premembrane and envelope proteins in a Venezuelan equine encephalitis (VEE) virus replicon particle (VRP) system was constructed and tested in conjunction with a plasmid DNA vaccine (D1ME-DNA) expressing identical dengue virus sequences. Cynomolgus macaques were vaccinated with three doses of DNA (DDD), three doses of VRP (VVV group), or a heterologous DNA prime-VRP boost regimen (DDV) using two doses of DNA vaccine and a third dose of VRP vaccine. Four weeks after the final immunization, the DDV group produced the highest dengue virus type 1-specific immunoglobulin G antibody responses and virus-neutralizing antibody titers. Moderate T-cell responses were demonstrated only in DDD- and DDV-vaccinated animals. When vaccinated animals were challenged with live virus, all vaccination regimens showed significant protection from viremia. DDV-immunized animals were completely protected from viremia (mean time of viremia = 0 days), whereas DDD- and VVV-vaccinated animals had mean times of viremia of 0.66 and 0.75 day, respectively, compared to 6.33 days for the control group of animals.