Evaluation of the reliability and accuracy of using cone-beam computed tomography for diagnosing periapical cysts from granulomas.

INTRODUCTION: The purpose of this study was to evaluate the reliability and accuracy of cone-beam computed tomographic (CBCT) imaging against the histopathologic diagnosis for the differential diagnosis of periapical cysts (cavitated lesions) from (solid) granulomas. METHODS: Thirty-six periapical lesions were imaged using CBCT scans. Apicoectomy surgeries were conducted for histopathological examination. Evaluator 1 examined each CBCT scan for the presence of 6 radiologic characteristics of a cyst (ie, location, periphery, shape, internal structure, effects on surrounding structure, and perforation of the cortical plate). Not every cyst showed all radiologic features (eg, not all cysts perforate the cortical plate). For the purpose of finding the minimum number of diagnostic criteria present in a scan to diagnose a lesion as a cyst, we conducted 6 receiver operating characteristic curve analyses comparing CBCT diagnoses with the histopathologic diagnosis. Two other independent evaluators examined the CBCT lesions. Statistical tests were conducted to examine the accuracy, inter-rater reliability, and intrarater reliability of CBCT images. RESULTS: Findings showed that a score of ≥4 positive findings was the optimal scoring system. The accuracies of differential diagnoses of 3 evaluators were moderate (area under the curve = 0.76, 0.70, and 0.69 for evaluators 1, 2, and 3, respectively). The inter-rater agreement of the 3 evaluators was excellent (α = 0.87). The intrarater agreement was good to excellent (κ = 0.71, 0.76, and 0.77). CONCLUSIONS: CBCT images can provide a moderately accurate diagnosis between cysts and granulomas.

Bacterial flora of dental periradicular lesions analyzed by the 454-pyrosequencing technology.

INTRODUCTION: Symptomatic teeth with periradicular lesions of infectious origin remain a significant challenge in dentistry, and the reason for the acute perturbation is incompletely understood. The present study used pyrosequencing of bacterial 16S ribosomal RNA (rRNA) genes to characterize the microbiota of periradicular lesions. METHODS: Thirteen periradicular lesions from 11 symptomatic and 2 asymptomatic teeth were sampled during apical surgery. Samples were subjected to DNA extraction and 16S rRNA polymerase chain reaction (PCR) amplification. PCR amplicons were then sequenced by using the Roche 454 GS FLX platform. Data were analyzed with the Quantitative Insights into Microbial Ecology (QIIME) software package. RESULTS: Seven of the 13 periradicular lesions (53.8%) yielded PCR amplicons, which generated 35,731 high-quality DNA sequences belonging to 10 bacterial phyla and 73 bacterial genera. All 7 lesions were associated with symptoms. The phyla with most bacterial taxa were Proteobacteria (proportion of total bacterial taxa, 33.3%), Firmicutes (30.9%), Actinobacteria (12.2%), and Bacteroidetes (11.4%). The most abundant genera were Fusobacterium (average of total sequences, 21.0%), Streptococcus (8.0%), Prevotella (7.5%), Corynebacterium (7.2%), Porphyromonas (6.0%). and Actinomyces (5.8%). CONCLUSIONS: This study demonstrated that the microbiota of symptomatic periapical lesions is predominated by anaerobic bacteria but also contains substantial levels of streptococci, actinomyces, and bacteria not previously identified in the oral cavity. The etiopathogenic role and therapeutic implication of periradicular bacteria need to be determined.

Metagenomic analysis of the viral flora of pine marten and European badger feces.

A thorough understanding of the diversity of viruses in wildlife provides epidemiological baseline information about potential pathogens. Metagenomic analysis of the enteric viral flora revealed a new anellovirus and bocavirus species in pine martens and a new circovirus-like virus and geminivirus-related DNA virus in European badgers. In addition, sequences with homology to viruses from the families Paramyxo- and Picornaviridae were detected.

Significance of human cytomegalovirus and Epstein-Barr virus in inducing cytokine expression in periapical lesions.

INTRODUCTION: Because herpesviruses might be etiologically involved in periapical pathosis of endodontic origin, this study aimed to determine the occurrence of human cytomegalovirus (HCMV), Epstein-Barr virus (EBV), and the expression of mRNA transcripts of tumor necrosis factor (TNF)-α, γ-interferon (IFN), interleukin (IL)-1ß, IL-6, IL-12, and IL-10 in periapical granulomatous lesions collected in conjunction with apicoectomy. METHODS: A total of 9 symptomatic and 6 asymptomatic teeth with periapical lesions were studied. Periapical samples were collected in conjunction with apicoectomy, which was being performed because of radiographic evidence of incomplete periapical healing after conventional root canal therapy. By using established polymerase chain reaction primers and procedures, polymerase chain reaction assays were used to identify herpesvirus and cytokine gene expression. RESULTS: The difference in occurrence of HCMV, EBV, and cytokines between symptomatic and asymptomatic periapical lesions was statistically significant: HCMV (P = .048), EBV (P = .002), IFN (P = .001), IL-1 (P = .012), IL-6 (P = .026), IL-10 (P = .026), IL-12 (P = .012), and TNF (P < .001) (Mann-Whitney U test). There was a significant correlation between EBV, HCMV, and TNF, γ-IFN, IL-1, and IL-12 in symptomatic periapical lesions (Spearman test). CONCLUSIONS: The present findings provide evidence of a putative role of HCMV and EBV in the pathogenesis of symptomatic periapical pathosis. The release of tissue-destructive cytokines might be of pathogenetic significance.

Genogroup I and II picobirnaviruses in respiratory tracts of pigs.

Sequence-independent amplification and specific reverse transcription PCRs identified genogroup I and II picobirnaviruses in respiratory tracts of pigs. These data expand knowledge of picobirnavirus diversity and tropism. Genetic relationships between porcine respiratory and human enteric picobirnaviruses suggest cross-species transmission of picobirnaviruses between pigs and humans.

Efficacy of the enamel matrix derivative to induce cementogenesis in vital and endodontically treated teeth with osseous dehiscence defects.

This experiment assessed the efficacy of the enamel matrix derivative (EMD) to regenerate cementum in vital and endodontically treated teeth with osseous dehiscence defects. Five adult female beagle dogs were used. Thirty maxillary teeth (bilateral maxillary canines and second and fourth premolars) were randomly divided into two experimental groups (groups A and B, containing 12 teeth each) and one control group (group C). Endodontic treatment was only performed on teeth in group A compared with teeth in groups B and C. Buccal osseous dehiscence defects were surgically created in teeth from all groups. Teeth in the experimental group were treated with the EMD, whereas the controls were not. After 5 months, the animals were sacrificed and block sections of the teeth in experimental and control groups were processed for histological analysis. Newly regenerated cementum was observed in all teeth in groups A and B. No cementum regeneration was observed in group C. There was a significant difference in cementum generation between the experimental and control groups (P < 0.001). EMD therapy induces cementogenesis in vital and endodontically treated teeth with osseous dehiscence defects.

Pulmonary pathology of pandemic influenza A/H1N1 virus (2009)-infected ferrets upon longitudinal evaluation by computed tomography.

We investigated the development of pulmonary lesions in ferrets by means of computed tomography (CT) following infection with the 2009 pandemic A/H1N1 influenza virus and compared the scans with gross pathology, histopathology and immunohistochemistry. Ground-glass opacities observed by CT scanning in all infected lungs corresponded to areas of alveolar oedema at necropsy. These areas were most pronounced on day 3 and gradually decreased from days 4 to 7 post-infection. This pilot study shows that the non-invasive imaging procedure allows quantification and characterization of influenza-induced pulmonary lesions in living animals under biosafety level 3 conditions and can thus be used in pre-clinical pharmaceutical efficacy studies.

Longevity of the protective immune response induced after vaccination with one or two doses of AS03A-adjuvanted split H5N1 vaccine in ferrets.

It is crucial that a safe and effective pandemic vaccine be rapidly available to combat a new pandemic threat. In this study we investigated the magnitude and persistence of the protective efficacy induced by one or two doses (3.75 µg HA/dose) of AS03(A)-adjuvanted H5N1 A/Indonesia/5/05 split vaccine in a lethal ferret challenge model. All ferrets that received at least one dose of adjuvanted vaccine 4 weeks before homologous challenge survived and showed reduced or undetectable virus replication in the lungs and the upper airways. Ferrets receiving two doses of adjuvanted vaccine 19 and 16 weeks before the challenge also showed high level of protection from replication in the lungs and the upper airways, albeit with only 83% survival. Animals in the control groups (non-adjuvanted vaccine or saline) and animals immunized with one dose of adjuvanted vaccine administered 10 or 16 weeks before challenge showed only 17-33% survival rate after challenge. In conclusion, our observations support the possibility that a single dose of AS03(A)-adjuvanted H5N1 split vaccine can offer a rapid and short term but partial protection against disease. A second dose of the adjuvanted vaccine, which can be given with a flexible injection schedule, was shown to be essential to induce appreciable levels of antibodies and long-term protection.

Pandemic H1N1 vaccine requires the use of an adjuvant to protect against challenge in naïve ferrets.

In the context of an A/H1N1 influenza pandemic situation, this study demonstrates that heterologous vaccination with an AS03-adjuvanted 2008/2009 seasonal trivalent and pandemic H5N1 monovalent split vaccine conferred partial protection in influenza-naïve ferrets after challenge with the influenza pandemic H1N1 A/The Netherlands/602/09 virus. Further, unlike saline control and non-adjuvanted vaccine, it was shown that immunization of naïve ferrets with an AS03-adjuvanted pandemic H1N1 A/California/7/09 influenza split vaccine induced increased antibody response and enhanced protection against the challenge strain, including significant reduction in viral shedding in the upper respiratory tract and reduced lung pathology post-challenge. These results show the need for vaccination with the adjuvanted vaccine to fully protect against viral replication and influenza disease in unprimed ferrets.

Identification and characterization of deer astroviruses.

The threat of emerging infectious viruses in humans requires a more effective approach regarding virus surveillance. A thorough understanding of virus diversity in wildlife provides epidemiological baseline information about pathogens and may lead to the identification of newly emerging pathogens in the future. In this study, diarrhoea samples from an outbreak of gastrointestinal illness in a Danish population of European roe deer were gathered for which no aetiological agent could be identified. Large-scale molecular RNA virus screening, based on host nucleic acid depletion, sequence-independent amplification and sequencing of partially purified viral RNA, revealed the presence of novel astroviruses, CcAstV-1 and CcAstV-2, in two of ten diarrhoea samples. Whether these viruses were responsible for causing diarrhoea remains to be determined. Phylogenetic analyses on amplified sequences showed that these viruses were most closely related to each other, were a novel species in the genus Mamastrovirus and may represent two different serotypes.

Human astrovirus infection in a patient with new-onset celiac disease.

Many diseases with unknown etiology may be caused by unidentified viruses. Sequence-independent amplification revealed a new astrovirus, similar to VA1, in a 4-year-old male diagnosed with celiac disease. This expands the geographic range of this virus to include Europe and may associate astrovirus infection with the onset of celiac disease.

Human picobirnaviruses identified by molecular screening of diarrhea samples.

The global threat of (re)emerging infectious viruses requires a more effective approach regarding virus surveillance and diagnostic assays, as current diagnostics are often virus species specific and not able to detect highly divergent or unknown viruses. A systematic exploration of viruses that infect humans is the key to effectively counter the potential public health threat caused by new and emerging infectious diseases. The human gut is a known reservoir of a wide variety of microorganisms, including viruses. In this study, Dutch clinical diarrhea samples for which no etiological agent could be identified by available cell culture, serological, or nucleic acid-based tests were gathered. Large-scale molecular RNA virus screening based on host nucleic acid depletion, sequence-independent amplification, and sequencing of partially purified viral RNA from a limited number of clinical diarrhea samples revealed four eukaryotic virus species. Among the detected viruses were a rhinovirus and a new picobirnavirus variant. In total, approximately 20% of clinical diarrhea samples contained human picobirnavirus sequences. The Dutch picobirnaviruses belonged to different phylogenetic clades and did not group with other picobirnaviruses according to year of isolation or host species. Interestingly, the average age of patients infected with picobirnavirus was significantly higher than that of uninfected patients. Our data show that sequence-independent amplification of partially purified viral RNA is an efficient procedure for identification of known and highly divergent new RNA viruses in clinical diarrhea samples.

MTA preparations from different origins may vary in their antimicrobial activity.

OBJECTIVE: The antimicrobial effects of 4 mineral trioxide aggregate (MTA) preparations, 2 white-colored (WMTA-1, WMTA-2) and 2 gray-colored (GMTA-1, GMTA-2), against C. albicans and E. faecalis were assessed in vitro. METHODOLOGY: Minimal inhibitory concentration (MIC) for each preparation was determined using the tube dilution test and Sabouraud agar media for C. albicans and brain heart infusion media for E. faecalis. Broth tubes were prepared and divided into experimental and control groups. Aliquots of each of the tested microorganisms were taken from a stock culture and added to each experimental and positive control group. All groups were incubated at 37 degrees C and evaluated for turbidity at 24-, 48-, and 72-hour time periods. Samples of 0.1 mL from each of the experimental and control tubes were subcultured on agar or brain heart infusion plates to confirm visible signs of bacterial or fungal growth. RESULTS: MIC of MTA against the 2 microorganisms tested varied among the 4 preparations tested. WMTA-1 and WMTA-2 inhibited C. albicans growth at concentrations of 3.125 mg/10 mL and 25 mg/10 mL, respectively, and statistically significant differences were found between WMTA-1 and WMTA-2 (P < .001). WMTA-1 and WMTA-2 inhibited E. faecalis growth at concentrations of 12.5 mg/10 mL and 50 mg/10 mL, respectively, and statistically significant differences were found between WMTA-1 and WMTA-2 (P < .001). GMTA-1 and GMTA-2 inhibited E. faecalis growth at concentrations of 12.5 mg/10 mL and 3.125 mg/10 mL, respectively, and statistically significant differences were found between GMTA-1 and GMTA-2 (P < .001). Both GMTA-1 and GMTA-2 inhibited C. albicans growth at a concentration of 3.125 mg/10 mL and no statistical differences were found between the preparations. Subculture of the broth tubes in agar or brain heart infusion plates confirmed the turbidity test result. CONCLUSION: The origin of MTA as well as the type of preparation may affect its antimicrobial characteristics. Clinicians should be aware of variations that may exist among such MTA preparations.

Successful treatment of a radicular groove by intentional replantation and Emdogain therapy: four years follow-up.

Radicular groove is an anatomical malformation that often causes severe periodontal defects. Treatments of such an anomaly present a clinical challenge to the operator. Presented is a case of successful treatment of radicular groove associated with a maxillary lateral incisor of a 15-year-old girl. A combination of endodontics, intentional replantation, and Emdogain therapy was used. At 4-year follow-up, the patient was comfortable and complete resolution of the periapical pathology was evident.

Cross-protection against lethal H5N1 challenge in ferrets with an adjuvanted pandemic influenza vaccine.

BACKGROUND: Unprecedented spread between birds and mammals of highly pathogenic avian influenza viruses (HPAI) of the H5N1 subtype has resulted in hundreds of human infections with a high fatality rate. This has highlighted the urgent need for the development of H5N1 vaccines that can be produced rapidly and in sufficient quantities. Potential pandemic inactivated vaccines will ideally induce substantial intra-subtypic cross-protection in humans to warrant the option of use, either prior to or just after the start of a pandemic outbreak. In the present study, we evaluated a split H5N1 A/H5N1/Vietnam/1194/04, clade 1 candidate vaccine, adjuvanted with a proprietary oil-in- water emulsion based Adjuvant System proven to be well-tolerated and highly immunogenic in the human (Leroux-Roels et al. (2007) The Lancet 370:580-589), for its ability to induce intra-subtypic cross-protection against clade 2 H5N1/A/Indonesia/5/05 challenge in ferrets. METHODOLOGY AND PRINCIPAL FINDINGS: All ferrets in control groups receiving non-adjuvanted vaccine or adjuvant alone failed to develop specific or cross-reactive neutralizing antibodies and all died or had to be euthanized within four days of virus challenge. Two doses of adjuvanted split H5N1 vaccine containing >or=1.7 microg HA induced neutralizing antibodies in the majority of ferrets to both clade 1 (17/23 (74%) responders) and clade 2 viruses (14/23 (61%) responders), and 96% (22/23) of vaccinees survived the lethal challenge. Furthermore lung virus loads and viral shedding in the upper respiratory tract were reduced in vaccinated animals relative to controls suggesting that vaccination might also confer a reduced risk of viral transmission. CONCLUSION: These protection data in a stringent challenge model in association with an excellent clinical profile highlight the potential of this adjuvanted H5N1 candidate vaccine as an effective tool in pandemic preparedness.

Clinical indications for digital imaging in dento-alveolar trauma. Part 1: traumatic injuries.

Traumatized teeth present a clinical challenge with regard to their diagnosis, treatment plan, and prognosis. Recent developments in imaging systems have enabled clinicians to visualize structural changes effectively. Computed tomography, magnetic resonance imaging and cone beam computed tomography are among the most commonly used systems for dental and maxillofacial surgery. The purpose of this review is to describe the advantages and disadvantages of each technique and the clinical application for dento-alveolar trauma. Three clinical cases are described to illustrate the potential use of the NewTom 3G for diagnosis and treatment plan of dento-alveolar traumatic injuries.

Clinical indications for digital imaging in dento-alveolar trauma. Part 2: root resorption.

Common complications of dento-alveolar trauma are pulp necrosis, pulp canal obliteration, periapical pathosis and root resorption. Different types of root resorption have been identified with traumatic injuries. Repair-related (surface), infection-related (inflammatory), ankylosis-related (osseous replacement) or extraradicular invasive cervical resorption are among the most common. Recent developments in imaging systems have enabled clinicians to visualize structural changes effectively. The diagnosis and three-dimensional imaging assessment of the resorption is important in order to determine the treatment complexity and expected outcome based on the location and extension of the root defect. This article discusses and illustrates the clinical application of cone beam computed tomography for diagnosis and treatment plan of root resorption. Four clinical cases are presented to illustrate the potential use of the NewTom 3G for root resorption.

Antibacterial effect of two mineral trioxide aggregate (MTA) preparations against Enterococcus faecalis and Streptococcus sanguis in vitro.

The antibacterial effects of gray-colored MTA (GMTA) and white-colored MTA (WMTA) against Enterococcus faecalis and Streptococcus sanguis were assessed in vitro using the tube dilution test. Broth tubes were prepared and divided into experimental and control groups. Aliquots of each of the tested microorganisms were taken from a stock culture and added to each experimental and positive control group. All groups were incubated at 37 degrees C and evaluated for turbidity at 0, 1, 24, 48, and 72-hour time periods. A direct correlation was found between GMTA and WMTA concentrations and their antibacterial effect. Tubes containing GMTA in concentrations of 50, 25, and 12.5 mg/ml did not show E. faecalis growth at any of the time periods tested whereas tubes containing WMTA showed E. faecalis growth at all concentrations and time periods tested. Statistically significant differences were found between tubes containing GMTA in concentrations of 50, 25 and 12.5 mg/ml and tubes containing similar concentrations of WMTA (p < 0.001). Tubes containing GMTA in concentrations of 50, 25, 12.5, 6.25, and 3.12 mg/ml and tubes containing WMTA in concentrations of 50, 25, and 12.5 mg/ml did not show S. sanguis growth at any of the time periods tested. Statistically significant differences were found between tubes containing GMTA in concentrations 6.25 and 3.12 mg/ml and tubes containing similar concentrations of WMTA (p < 0.001). It appears that the susceptibility of E. faecalis and S. sanguis to MTA differed and that GMTA requires lower concentrations than WMTA to exert the same antibacterial effect against each of the microorganisms tested.

Differential diagnosis of large periapical lesions using cone-beam computed tomography measurements and biopsy.

This study compared the differential diagnosis of large periapical lesions (granuloma versus cyst) to traditional biopsy using Cone Beam Computed Tomography (CBCT), the NewTom 3G. Large lesions were scanned and a preoperative diagnosis based on gray value measurements of the imaged lesion area was made. After surgery a biopsy report was obtained and compared to the CBCT diagnosis. In 13 out of 17 cases, the diagnosis coincided. In 4 out of 17 cases, the CBCT read cyst with the oral pathologist's diagnosis being granuloma. Thus, the CBCT may provide a more accurate diagnosis than biopsy and histology providing a diagnosis without invasive surgery and/or waiting a year to see if nonsurgical therapy is effective.