Musculoaponeurotic architecture of the human masseter muscle: an in vivo ultrasonographic study of architectural changes during mandibular protrusion and lateral excursions.

OBJECTIVE: The present study evaluated the in vivo musculoaponeurotic architecture of the superficial head (SH) of the masseter muscle (MM) of asymptomatic participants in excursive mandibular movements compared to the relaxed state as examined with ultrasonography. It was hypothesized that the mean fiber bundle length (FBL) and mean height of the aponeurosis (HA) of the laminae of the SH would differ significantly between the relaxed state and protrusion, ipsilateral excursion, and contralateral excursion. STUDY DESIGN: The MM was studied volumetrically in 12 female and 12 male asymptomatic participants bilaterally by using ultrasound imaging. Mean FBL and HA in protrusion and ipsilateral and contralateral excursion were compared to these values in the relaxed state using paired t tests (P < .05). The intraclass correlation coefficient was used to assess intraexaminer reliability. RESULTS: The SH exhibited multiple laminae. Fiber bundles were found to attach to bone and the superior and inferior aponeuroses. Mean FBL was significantly shorter and mean HA significantly longer in protrusion and the excursions than in the relaxed state although the pattern of altered laminae and aponeuroses differed among the mandibular movements. Intraexaminer reliability was excellent. CONCLUSION: Specific changes in mean FBL and mean HA suggest differential contraction of the SH of the MM based on laminar morphology. These findings provide a baseline to investigate musculoaponeurotic changes in patients with myogenic masseter muscle pain.

Detection of PSMA expression on circulating tumor cells by blood-based liquid biopsy in prostate cancer.

Background: For patients with mCRPC, PSMA-targeted radioligand treatment has significantly improved the clinical outcome. A blood-based liquid biopsy assay for recognizing PSMA protein expression on circulating tumor cells may be beneficial for better informing therapeutic decision-making and identifying the patients most likely to benefit from PSMA-targeted radioligand therapy. Methods: Using high-throughput imaging and digital AI pathology algorithms, a four-color immunofluorescence assay has been developed to find PSMA protein expression on CTCs on a glass slide. Cell line cells (LNCaP/PC3s/22Rv1) spiked into healthy donor blood were used to study the precision, specificity, sensitivity, limit of detection, and overall accuracy of the assay. Clinical validation and low-pass whole-genome sequencing were performed in PSMA-PET-positive patients with high-risk mCRPC (N = 24) utilizing 3 mL of blood. Results: The PSMA CTC IF assay achieved analytical specificity, sensitivity, and overall accuracy above 99% with high precision. In the clinical validation, 76% (16/21) of the cases were PSMA positive with CTC heterogeneity, and 88% (21/24) of the patients contained at least one conventional CTC per milliliter of blood. Thirty-six low-pass-sequenced CTCs from 11 individuals with mCRPC frequently exhibited copy number increases in AR and MYC and losses in RB1, PTEN, TP53, and BRCA2 locus. Conclusions: The analytical validation utilizing Epic Sciences' liquid biopsy CTC platform demonstrated the potential to detect PSMA protein expression in CTCs from patients with mCRPC. This assay is positioned as an effective research tool to evaluate PSMA expression, heterogeneity, and therapeutic response in many ongoing clinical studies to target tumors that express PSMA.

Optimizing effective dose and image quality in cone beam CT sialography.

OBJECTIVES: The purpose of this study was to develop protocols that optimize patient radiation dose and image quality for cone beam computed tomographic (CBCT) sialography for the major salivary glands. STUDY DESIGN: Radiation absorbed dose measurements were repeated in triplicate using 25 sites in the head and neck of a Radiation ANalog DOsimetry system (RANDO) phantom, and effective doses were calculated across a range of peak kilovoltage (kVp) and milliamperage (mA) settings using an 8 cm (diameter) by 5 cm (height) field of view (FOV) for submandibular imaging and an 8 cm (diameter) by 8 cm (height) FOV for parotid imaging. Image signal difference-to-noise ratio (SDNR) was determined, and the figure-of-merit (FOM), a measure of image quality, was calculated. RESULTS: For submandibular sialography, 85 kVp and 6 mA were chosen as the optimal exposure parameters, resulting in a mean effective dose of 82.47 µSv and a mean SDNR of 13.86, with a mean FOM of 2.33 µSv-1. For parotid sialography, 70 kVp and 6 mA were chosen, and these settings resulted in a mean effective dose of 39.99 µSv, a mean SDNR of 17.43, and a mean FOM of 7.60 µSv-1. CONCLUSIONS: Low-dose 3-dimensional sialography with high image quality and minimal effective dose can be delivered using CBCT with localized, small FOVs.

Analysis of BRCA2 Copy Number Loss and Genomic Instability in Circulating Tumor Cells from Patients with Metastatic Castration-resistant Prostate Cancer.

BACKGROUND: BRCA2 alterations predict for a response to poly-ADP-ribose polymerase inhibition in metastatic castration-resistant prostate cancer (mCRPC). However, detection is hindered by insufficient tumor tissue and low sensitivity of cell-free DNA for detecting copy number loss. OBJECTIVE: To evaluate the BRCA2 loss detection using single-cell, shallow whole-genome sequencing (sWGS) of circulating tumor cells (CTCs) in patients with mCRPC. DESIGN, SETTING, AND PARTICIPANTS: We analyzed CTC samples collected concurrently with tumor biopsies intended for clinical sequencing in patients with progressing mCRPC. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: Differences in proportions were evaluated using the chi-square test. Correlations between assays were analyzed in linear regression models. Associations between alterations and genomic instability were assessed on the single-cell level using mixed-effect negative binomial models. RESULTS AND LIMITATIONS: We identified 138 patients with concurrent CTC and biopsy samples. CTC sWGS generated copy number profiles in a similar proportion of patients to biopsy samples (83% vs 78%, p = 0.23), but was more effective than bone biopsies (79% vs 50%; p = 0.009). CTC sWGS detected BRCA2 loss in more patients than tissue at the ≥1 (42% vs 16%; p < 0.001) and ≥2 (27% vs 16%; p = 0.028) CTC thresholds. The overall prevalence of BRCA2 loss was not increased in CTCs using sample-level composite z scores (p = 0.4), but was significantly increased compared with a lower-than-expected prevalence in bone samples (21% vs 3%, p = 0.014). Positive/negative predictive values for CTC BRCA2 loss were 89%/96% using the ≥1 CTC threshold and 67%/92% using the composite z score. CTC BRCA2 loss was associated with higher genomic instability in univariate (1.4-fold large-scale transition difference, 95% confidence interval [CI]: 1.2-1.6; p < 0.001) and multivariable analysis (1.4-fold difference, 95% CI: 1.2-1.6; p < 0.001). CONCLUSIONS: Copy number profiles can reliably be generated using CTC sWGS, which detected a majority of tissue-confirmed BRCA2 loss and "CTC-only" losses. BRCA2 losses were supported by increases in genomic instability. PATIENT SUMMARY: Current testing strategies have limitations in their ability to detect BRCA2 loss, a relatively common alteration in prostate cancer that is used to identify patients who may benefit from targeted therapy. In this paper, we evaluated whether we could detect BRCA2 loss in individual tumor cells isolated from patient blood samples and found this method to be suitable for further analysis.

Interexaminer Agreement in the Radiologic Identification of Apical Periodontitis/Rarefying Osteitis in the National Dental Practice-Based Research Network PREDICT Endodontic Study.

INTRODUCTION: Periapical images are routinely made in endodontics to support diagnosis and treatment decisions, but conventional imaging may not readily demonstrate inflammatory changes. This study aims to quantify disagreement in the radiologic interpretation of apical periodontitis/rarefying osteitis between 2 expert examiners and to determine if differences exist based on anatomic location. METHODS: We used 1717 pretreatment periapical images made before orthograde endodontic treatment as part of the Predicting Outcomes of Root Canal Treatment (PREDICT) study conducted within the National Dental Practice-Based Research Network. Periapical changes were assessed independently by 2 board-certified specialists, an oral and maxillofacial radiologist and an endodontist, blinded to other clinical information. If the examiners disagreed about whether a diagnosis of apical periodontitis/rarefying osteitis was justified, an adjudication was made by a third examiner. RESULTS: The overall prevalence of this radiologic diagnosis in the periapical images was 55%, and interexaminer agreement measured with the Cohen kappa statistic was calculated to be 0.56 (95% confidence interval, 0.52-0.60). Diagnostic disagreements between the 2 examiners occurred for 377 teeth (22%), with disagreements more frequent for jaw location (P = .038) and tooth type (P = .021). Differences between root number (P = .058) and jaw location and tooth groups (P = .069) were found not to be statistically significant. CONCLUSIONS: The variability of diagnostic disagreements across anatomic location and tooth type may reflect the inability of periapical images to reveal bone changes masked by the complexity and density of overlying anatomic structures, a limitation that could potentially be overcome with the use of 3-dimensional imaging.

Variation of the response to metal ions and nonsense-mediated mRNA decay across different Saccharomyces cerevisiae genetic backgrounds.

Regulation of mRNA steady-state levels is important in controlling gene expression particularly in response to environmental stimuli. This allows cells to rapidly respond to environment changes. The highly conserved nonsense-mediated mRNA decay (NMD) pathway was initially identified as a pathway that degrades aberrant mRNAs. NMD is now recognized as a pathway with additional functions including precisely regulating the expression of select natural mRNAs. Majority of these natural mRNAs encode fully functional proteins. Regulation of natural mRNAs by NMD is activated by NMD targeting features and environmental cues. Here, we show that Saccharomyces cerevisiae strains from three genetic backgrounds respond differentially to NMD depending on the environmental stimuli. We found that wild type and NMD mutant W303a, BY4741, and RM11-1a yeast strains respond similarly to copper in the environment but respond differentially to toxic cadmium. Furthermore, the PCA1 alleles encoding different mRNAs from W303a and RM11-1a strains are regulated similarly by NMD in response to the bio-metal copper but differentially in response to toxic cadmium.

A dynamic ultrasonographic in vivo study of the musculoaponeurotic architecture of the human masseter muscle.

OBJECTIVE: In the present study, we sought to investigate the in vivo musculoaponeurotic architecture of the masseter muscle (MM) volumetrically with ultrasound in asymptomatic participants. It was hypothesized that the mean fiber bundle length (FBL) and mean aponeurotic height of laminae of the MM differ significantly between the relaxed state and maximally contracted state upon elevation of the mandible. STUDY DESIGN: The MM was investigated volumetrically in 12 male and 12 female asymptomatic participants (mean age, 25.8 ± 4.1 years) using ultrasound. The mean FBL and mean height of aponeuroses in the relaxed and maximally contracted states were compared using paired t tests, with significance established at P ≤ .05. Intrarater reliability was assessed using the intraclass correlation coefficient (ICC). RESULTS: The MM consisted of the superficial head (SH) and deep head, each arranged in multiple laminae. Fiber bundles extended between superior and inferior aponeuroses and/or bone. Statistically significant differences (P ≤ .05) were observed in mean FBL and in mean height of aponeuroses between the relaxed and contracted states only in superficial laminae of the SH. CONCLUSIONS: These results suggest there is differential contraction of the laminae of the MM in the transition from relaxed to contracted states. Future comparison with pathologic patients can be made on the basis of an established normative database.

Radiograph prescription practices of dentists in Ontario, Canada.

BACKGROUND: With the advent of new ionizing radiation technologies, consideration must be given to the use of radiography in clinical practice. The authors aimed to determine how general dentists obtain oral and maxillofacial images and whether dentists' demographic characteristics and practice environments affect imaging decisions. METHODS: The authors conducted a cross-sectional study using an electronic survey to collect information from general dentists in Ontario, Canada's most populous province. RESULTS: The authors received 1,332 responses from 9,052 general dentists registered to practice in Ontario. A total of 57.2% of dentists used a nonindividualized approach to obtaining images for new adult dentate patients. Using clinical case scenarios, the authors found that 18.8% through 55.1% of dentists did not follow evidence-based guidelines when obtaining images. Overprescribing imaging was 5.8 times more common than underprescribing. Seeing patients after images were obtained (odds ratio [OR], 5.5), owning a panoramic and a cone-beam computed tomography imaging system (OR, 4.4), and the belief that images should be obtained to screen for occult pathoses (OR, 3.5) were the factors most strongly associated with overprescription. CONCLUSIONS: Radiography prescription practices varied considerably among general dentists in Ontario and did not conform to established guidelines. Factors other than patient needs appeared to influence imaging practices. PRACTICAL IMPLICATIONS: Overprescription of imaging by general dentists can result in unnecessary radiation exposure and financial costs to patients. To mitigate this, changes are needed in dental education, continuing education courses, and regulatory requirements.

Effectiveness of Commercial Software-Enhanced Image Artifact Reduction Software.

INTRODUCTION: Artifact reduction (AR) software has been incorporated into some cone-beam computed tomographic (CBCT) systems to reduce the severity of beam hardening (BH) artifacts and improve image quality. This study quantifies BH artifact and evaluates the effectiveness of AR in 2 CBCT systems. METHODS: Palatal roots of Dent-Alike (Dentsply Sirona, Tulsa, OK) teeth were prepared and root filled with gutta-percha and EndoSequence BC Sealer (Brasseler, Savannah, GA). Six teeth were imaged with and without AR software using the ProMax3D (Planmeca Oy, Helsinki, Finland) and the Pax-i3D (Vatech, Hwaseong-si, South Korea) systems. FSL (FMRIB, Oxford, UK) software was used to quantify the light and dark components of the BH artifact along the tooth root using a specific region of interest approach and an image-wide analysis approach. Statistical analysis was performed using paired t tests and corrected for multiple comparisons with cluster mass correction using a nonparametric statistical analysis to evaluate the differences in the artifact volumes and areas with and without AR. RESULTS: A significant reduction in the light artifact was observed with the Planmeca system (P < .05), but no significant differences were observed for either the light or dark artifacts with the Vatech system when AR was applied. There were also significant reductions in the volumes of light and dark artifacts along the entire root length when the AR was applied with the Planmeca system (cluster mass P < .05), but no significant differences were observed with the Vatech system. CONCLUSIONS: Proprietary AR software is not equally effective in reducing the light and/or dark components of CBCT BH artifacts.

The use of radiographic imaging technologies by general dentists in Ontario, Canada.

OBJECTIVE: The aim of this study was to determine the prevalence of 2-dimensional and 3-dimensional digital imaging technologies, the methods used by general dentists to limit patient exposure to ionizing radiation, and the impact of dentists' education on imaging technologies and patient dose-reducing techniques. STUDY DESIGN: A cross-sectional, web-based survey of all general dentists in Ontario was conducted. RESULTS: Responses from 1332 (14.7%) of the 9052 registered general dentists in Ontario were included in the analysis. Approximately 89% reported using digital intraoral technology, 81.1% reported owning panoramic imaging systems, 71.2% reported making referrals for cone beam computed tomography (CBCT), and 9.5% reported including CBCT in their practices. CBCT was most commonly used for dental implant treatment planning (85.8%), followed by endodontics (45.4%), evaluation of pathology (39.6%), and surgical assessment for impacted teeth or difficult extractions (36.8%). Approximately 32.7% used only collimators with a long focal point-receptor distance and 8% used only rectangular collimation; 86.9% reported using a thyroid collar when imaging patients. Differences in educational backgrounds correlated with differences in the use of imaging and dose-reducing techniques. CONCLUSIONS: There is widespread adoption of digital imaging technologies by general dentists in Ontario, including CBCT. Greater implementation of long and/or rectangular collimation could markedly reduce the ionizing radiation dose to patients. Changes in dental education curricula and continuing education course offerings may address these issues.

Structural abnormalities in the temporalis musculo-aponeurotic complex in chronic muscular temporomandibular disorders.

Some forms of chronic pain are thought to be driven and maintained by nociceptive input, which can drive plasticity within nociceptive pathways. We have previously identified abnormalities along the entire nociceptive pathway in chronic myalgic temporomandibular disorders (mTMD), including the trigeminal nerves, brainstem pathways, and in the thalamus and somatosensory cortex. These data suggest that there is a peripheral nociceptive drive in mTMD, but the source of this nociceptive activity remains unknown. Here, our aim was to determine whether structural abnormalities exist in the muscles of mastication of patients with chronic mTMD. Specifically, we tested whether the volume of the temporalis muscle and its tendon-aponeurosis complex (TAC, a structure that dissipates forces in a muscle) in mTMD patients differ compared to age- and sex-matched controls. To do so, we segmented these structures on T1-weighted structural magnetic resonance images. We found that muscle volumes in mTMD were not different to controls. However, the mTMD group had significantly smaller volumes of the bilateral temporalis TAC, and thus a smaller TAC-to-muscle volume ratio. These findings were consistent across 2 independent cohorts of 17 mTMD patients, compared to 17 age- and sex-matched controls. We propose a model where reduced TAC-to-muscle ratio could result in a predisposition to muscle tissue injury. In sum, abnormalities of the temporalis muscles in mTMD supports our hypothesis that chronic mTMD pathophysiology may be related to peripheral nociceptive barrage originating from the muscles of mastication.

Effectiveness of overnight learning commons: A comparative study.

This study explores the overnight usage of Chi Wah Learning Commons and The University of Hong Kong Libraries on the same campus. By adopting quantitative research, it investigates students' preferences in these areas and levels of social capital in the library, further, it explores the similarities and differences of user behavior between the university's undergraduate and postgraduate students. Our findings showed that HKUL had not taken full advantage of its prime location and comprehensive collection in the main library, and the full potential of overnight services. The results of this study contribute to the library management in understanding the changing needs of users and propose the potential enhancements on social capital among students and the community.

Multi-platform discovery of haplotype-resolved structural variation in human genomes.

The incomplete identification of structural variants (SVs) from whole-genome sequencing data limits studies of human genetic diversity and disease association. Here, we apply a suite of long-read, short-read, strand-specific sequencing technologies, optical mapping, and variant discovery algorithms to comprehensively analyze three trios to define the full spectrum of human genetic variation in a haplotype-resolved manner. We identify 818,054 indel variants (<50 bp) and 27,622 SVs (≥50 bp) per genome. We also discover 156 inversions per genome and 58 of the inversions intersect with the critical regions of recurrent microdeletion and microduplication syndromes. Taken together, our SV callsets represent a three to sevenfold increase in SV detection compared to most standard high-throughput sequencing studies, including those from the 1000 Genomes Project. The methods and the dataset presented serve as a gold standard for the scientific community allowing us to make recommendations for maximizing structural variation sensitivity for future genome sequencing studies.

Long-read single-molecule maps of the functional methylome.

We report on the development of a methylation analysis workflow for optical detection of fluorescent methylation profiles along chromosomal DNA molecules. In combination with Bionano Genomics genome mapping technology, these profiles provide a hybrid genetic/epigenetic genome-wide map composed of DNA molecules spanning hundreds of kilobase pairs. The method provides kilobase pair-scale genomic methylation patterns comparable to whole-genome bisulfite sequencing (WGBS) along genes and regulatory elements. These long single-molecule reads allow for methylation variation calling and analysis of large structural aberrations such as pathogenic macrosatellite arrays not accessible to single-cell second-generation sequencing. The method is applied here to study facioscapulohumeral muscular dystrophy (FSHD), simultaneously recording the haplotype, copy number, and methylation status of the disease-associated, highly repetitive locus on Chromosome 4q.

Genome maps across 26 human populations reveal population-specific patterns of structural variation.

Large structural variants (SVs) in the human genome are difficult to detect and study by conventional sequencing technologies. With long-range genome analysis platforms, such as optical mapping, one can identify large SVs (>2 kb) across the genome in one experiment. Analyzing optical genome maps of 154 individuals from the 26 populations sequenced in the 1000 Genomes Project, we find that phylogenetic population patterns of large SVs are similar to those of single nucleotide variations in 86% of the human genome, while ~2% of the genome has high structural complexity. We are able to characterize SVs in many intractable regions of the genome, including segmental duplications and subtelomeric, pericentromeric, and acrocentric areas. In addition, we discover ~60 Mb of non-redundant genome content missing in the reference genome sequence assembly. Our results highlight the need for a comprehensive set of alternate haplotypes from different populations to represent SV patterns in the genome.

Structural Variation Detection and Analysis Using Bionano Optical Mapping.

The need to accurately identify the complete structural variation profile of genomes is becoming increasingly evident. In contrast to reference-based methods like sequencing or comparative methods like aCGH, optical mapping is a de novo assembly-based method that enables better realization of true genomic structure. It allows for independently detecting balanced and unbalanced structural variants (SVs) from separate alleles and for discovering de novo events. Here we show how Bionano Genome Mapping creates de novo assemblies from native and intact, megabase-scale DNA molecules and uses those assemblies to detect a wide range of structural variants.

High-resolution comparative analysis of great ape genomes.

Genetic studies of human evolution require high-quality contiguous ape genome assemblies that are not guided by the human reference. We coupled long-read sequence assembly and full-length complementary DNA sequencing with a multiplatform scaffolding approach to produce ab initio chimpanzee and orangutan genome assemblies. By comparing these with two long-read de novo human genome assemblies and a gorilla genome assembly, we characterized lineage-specific and shared great ape genetic variation ranging from single- to mega-base pair-sized variants. We identified ~17,000 fixed human-specific structural variants identifying genic and putative regulatory changes that have emerged in humans since divergence from nonhuman apes. Interestingly, these variants are enriched near genes that are down-regulated in human compared to chimpanzee cerebral organoids, particularly in cells analogous to radial glial neural progenitors.

A Novel Method for Characterizing Beam Hardening Artifacts in Cone-beam Computed Tomographic Images.

INTRODUCTION: The beam hardening (BH) artifact produced by root filling materials in cone-beam computed tomographic (CBCT) images is influenced by their radiologic K absorption edge values. The purpose of this study was to describe a novel technique to characterize BH artifacts in CBCT images produced by 3 root canal filling materials and to evaluate the effects of a zirconium (Zr)-based root filling material with a lower K edge (17.99 keV) on the production of BH artifacts. METHODS: The palatal root canals of 3 phantom model teeth were prepared and root filled with gutta-percha (GP), a Zr root filling material, and calcium hydroxide paste. Each phantom tooth was individually imaged using the CS 9000 CBCT unit (Carestream, Atlanta, GA). The "light" and "dark" components of the BH artifacts were quantified separately using ImageJ software (National Institutes of Health, Bethesda, MD) in 3 regions of the root. Mixed-design analysis of variance was used to evaluate differences in the artifact area for the light and dark elements of the BH artifacts. RESULTS: A statistically significant difference in the area of the dark portion of the BH artifact was found between all fill materials and in all regions of the phantom tooth root (P < .05). GP generated a significantly greater dark but not light artifact area compared with Zr (P < .05). Moreover, statistically significant differences between the areas of both the light and dark artifacts were observed within all regions of the tooth root, with the greatest artifact being generated in the coronal third of the root (P < .001). CONCLUSIONS: Root canal filling materials with lower K edge material properties reduce BH artifacts along the entire length of the root canal and reduce the contribution of the dark artifact.

The Performance of a Zirconium-based Root Filling Material with Artifact Reduction Properties in the Detection of Artificially Induced Root Fractures Using Cone-beam Computed Tomographic Imaging.

INTRODUCTION: Limited field of view cone-beam computed tomographic (CBCT) imaging has been used to augment clinical testing of vertical root fractures (VRFs); however, the presence of gutta-percha (GP) in the canal space generates substantial imaging artifacts that make fracture detection difficult. The purpose of this study was to evaluate the influence of a zirconium (Zr)-based root filling material with radiologic properties that reduce beam hardening (BH) artifacts using CBCT imaging in the in vitro diagnosis of VRFs. METHODS: One hundred seventy-six single-rooted mandibular premolar teeth were obtained, and half of these teeth were filled with GP or Zr (CPoint; EndoTechnologies, LLC, Shrewsbury, MA). VRFs were induced in 44 decoronated teeth in each group using an Instron (Norwood, MA) Universal Testing Machine. Each root was then placed in a dry human mandible and imaged with the Carestream 9000 3D CBCT system (Carestream Dental, Atlanta, GA). The images were evaluated by 6 oral maxillofacial radiologists (OMRs) and residents. RESULTS: The sensitivity was greater for detecting VRFs in the Zr group than the GP group (P = .035). However, the specificity was greater for the GP group than the Zr group (P = .028). Receiver operating characteristic area under the curve values were greater for the Zr group than the GP group, but these differences were not statistically significant. The OMRs outperformed the residents in the detection of VRFs in the Zr group with respect to specificity (P = .006) and positive predictive value (P = .012). CONCLUSIONS: The reduced BH of the Zr group improved the sensitivity of the detection of artificially induced VRFs. The ability to detect VRFs in the Zr group was further enhanced by clinical experience.