Oral cancer detection and progression prediction using noninvasive cytology-based DNA ploidy approach.

BACKGROUND: Despite the oral cavity being readily accessible, oral cancer (OC) remains a significant burden. The objective of this study is to develop a DNA ploidy-based cytology test for early detection of high-risk oral lesions. METHODS: This retrospective study was conducted using 569 oral brushing samples collected from 95 normal and 474 clinically abnormal mucosa with biopsy diagnosis of reactive, low-grade or high-grade precancer or cancers. Brushing cells were processed to characterize DNA ploidy. A two-step DNA ploidy-based algorithm, the DNA ploidy oral cytology (DOC) test, was developed using a training set, and verified in test and validation sets to differentiate high-grade lesions (HGLs) from normal. The prognostic value of the test was evaluated by an independent outcome cohort, including progressed and non-progressing normal, reactive and low-grade lesions. Classification performance was assessed by accuracy, sensitivity, and specificity, while the prognostic value was evaluated by using the Cox proportional hazards analysis on 3-year progression-free survival (PFS). RESULTS: The developed DOC test exhibited high accuracy for detecting HGLs in the test and validation sets, with a sensitivity of 0.97 and 0.96, respectively. Its application to the Outcome cohort demonstrated significant prognostic value for 3-year PFS (log rank, p < 0.001). Multivariate analysis showed that high-grade pathology was the only variable explaining positive DOC test, not age, smoking, or lesional site. CONCLUSION: Clinical implementation of the DOC test could provide an effective screening method for detecting HGLs for biopsy and lesions at risk of progression.

Three-Dimension Epithelial Segmentation in Optical Coherence Tomography of the Oral Cavity Using Deep Learning.

This paper aims to simplify the application of optical coherence tomography (OCT) for the examination of subsurface morphology in the oral cavity and reduce barriers towards the adoption of OCT as a biopsy guidance device. The aim of this work was to develop automated software tools for the simplified analysis of the large volume of data collected during OCT. Imaging and corresponding histopathology were acquired in-clinic using a wide-field endoscopic OCT system. An annotated dataset (n = 294 images) from 60 patients (34 male and 26 female) was assembled to train four unique neural networks. A deep learning pipeline was built using convolutional and modified u-net models to detect the imaging field of view (network 1), detect artifacts (network 2), identify the tissue surface (network 3), and identify the presence and location of the epithelial-stromal boundary (network 4). The area under the curve of the image and artifact detection networks was 1.00 and 0.94, respectively. The Dice similarity score for the surface and epithelial-stromal boundary segmentation networks was 0.98 and 0.83, respectively. Deep learning (DL) techniques can identify the location and variations in the epithelial surface and epithelial-stromal boundary in OCT images of the oral mucosa. Segmentation results can be synthesized into accessible en face maps to allow easier visualization of changes.

Fecal Microbiota, Live-jslm for the Prevention of Recurrent Clostridioides difficile Infection : Subgroup Analysis of PUNCH CD2 and PUNCH CD3.

GOALS: To assess fecal microbiota, live-jslm (REBYOTA, abbreviated as RBL, formerly RBX2660) efficacy and safety in participants grouped by recurrent Clostridioides difficile infection (rCDI) risk factors and treatment-related variables. BACKGROUND: RBL is the first microbiota-based live biotherapeutic approved by the US Food and Drug Administration for the prevention of rCDI in adults after antibiotic treatment for rCDI. STUDY: Treatment success rates across subgroups for PUNCH CD3 (NCT03244644) were estimated using a Bayesian hierarchical model, borrowing data from PUNCH CD2 (NCT02299570). Treatment-emergent adverse events were summarized for the double-blind treatment period within 8 weeks. RESULTS: Treatment differences between RBL and placebo at 8 weeks were similar to the total population for most subgroups. Treatment effect sizes were similar between CDI tests, higher for oral vancomycin courses >14 days versus ≤14 days and higher for antibiotic washout periods of 3 days versus ≤2 days. The largest reductions in the rate of rCDI with RBL versus placebo were observed for participants with a 3-day CDI antibiotic washout period and participants with ≥4 previous CDI episodes. Most RBL-treated participants experienced TEAEs that were mild or moderate in severity and related to preexisting conditions. CONCLUSION: This analysis provides further evidence of RBL efficacy and safety across subgroups, including those at high risk for rCDI.

Microbiota-Based Live Biotherapeutic RBX2660 for the Reduction of Recurrent Clostridioides difficile Infection in Older Adults With Underlying Comorbidities.

Background: Advanced age and underlying comorbidities are associated with greater rates of recurrence in patients with Clostridioides difficile infection (CDI). Reducing the likelihood of recurrence through treatment with an antimicrobial followed by a microbiota replacement therapy can decrease the burden of this infection and improve patient outcomes. We report the efficacy and safety of RBX2660, a microbiota-based live biotherapeutic, in older adults with recurrent CDI, grouped by comorbidities. Methods: In this post hoc subgroup analysis of the PUNCH CD3 trial, we assessed outcomes in older adults (age ≥65 years) grouped by Charlson Comorbidity Index severity scores at screening (moderate [3-4] and severe [≥5]) and by the presence of underlying cardiac, renal, or gastrointestinal disorders. Results: RBX2660 treatment success rates in older adults with comorbidities were consistent across subgroups and similar to those in the total RBX2660-treated population. A greater percentage of RBX2660-treated older adults remained free of CDI recurrence through 8 weeks following treatment compared with placebo-treated participants in all but 2 subgroups assessed. Across all subgroups, most treatment-emergent adverse events (TEAEs) were mild or moderate in severity and related to a preexisting condition. None of the serious or life-threatening TEAEs that occurred were related to RBX2660 or its administration. Occurrence of TEAEs did not cluster in any subgroup. Conclusions: RBX2660 is efficacious and safe in older adults with recurrent CDI and underlying comorbidities.

Separation of Emulsions with Fibrous Filter-Coalescers.

During froth treatment, a water-in-diluted bitumen emulsion is obtained. The emulsified water contains chloride ions that form hydrochloric acid in downstream oil processing, leading to catalyst deactivation and equipment corrosion. Emulsified water, drops smaller than 10 µm, cannot be effectively removed by gravity settling and centrifugation to below 2 wt %. In this work, a filter-coalescer was used to promote the coalescence and separation of water-in-bitumen emulsion. The larger water drops (>300 µm) exiting the coalescer undergo gravity settling, reducing the water content in diluted bitumen emulsions to values lower than 0.1 vol %. The performance of the coalescer was interpreted via the colloid filtration theory of Rajagopalan and Tien (RT), improved in this work with a coalescence probability (CP) prefactor. This new RTCP framework was able to reproduce the experimental data, allowing its potential use as a predictive model for emulsion filtration and the operation of filter-coalescers. A capillary number analysis was used to account for the detachment of coalesced drops and interpret the drop sizes with different superficial velocities and bed porosities.

Review of Treatment Options for Irritable Bowel Syndrome with Constipation and Chronic Idiopathic Constipation.

Irritable bowel syndrome with constipation (IBS-C) and chronic idiopathic constipation (CIC) are two common disorders of gut-brain interaction. Affected patients often first present to their primary care providers seeking care for symptoms of constipation, abdominal pain, and bloating, which have a significant impact on their health-related quality of life. These patients often require extensive counseling and reassurance, and knowledge of reliable diagnostic criteria and treatment options is imperative to managing their conditions. Family medicine practitioners, including nurse practitioners and physician assistants, are uniquely qualified to provide a diagnosis and safe, effective management of these disorders. This article reviews the latest evidence and provides practical advice related to diagnosis and management of IBS-C and CIC.

Interfacial Tension of the Water-Diluted Bitumen Interface at High Bitumen Concentrations Measured Using a Microfluidic Technique.

The interfacial tension (IFT) is a critical parameter to inform our understanding of the phenomena of drop breakup and droplet-droplet coalescence in sheared water-in-diluted bitumen (dilbit) emulsions. A microfluidic extensional flow device (MEFD) was used to determine the IFT of the dilbit-water emulsion system for bitumen concentrations of 33%, 50%, and 67% by weight (solvent to bitumen ratio (S/B) = 2, 1, and 0.5, respectively) and two different pH values of water: 8.3 and 9.9. The IFT was observed to increase with the bitumen concentration and decrease significantly upon lowering the water pH. The time scale for achieving the steady state IFT increased with bitumen concentration and was less sensitive to the water pH. But the most important feature of our measurements is that the IFTs recorded were significantly smaller than the values reported in the literature. We recognized two important differences between our studies and prior investigations: measurement of the IFT of water drops in dilbit as opposed to dilbit drops in water in earlier studies, and time scales of measurement of IFT that ranged from hundreds of milliseconds to a few seconds, as compared to a minute or longer in past investigations. These differences were examined carefully, but neither was found to explain the low IFTs measured in our studies. Our work leads to the following hypothesis: the mechanical properties of the interface of a sheared water drop in bitumen are significantly different from a stagnant one.

Oral health status and possible explanatory factors of an inner-city low-income community.

BACKGROUND/PURPOSE: Individuals with low income bear a number of health challenges to healthcare services. Vancouver's Downtown Eastside (DTES) is known to be a low-income community in a metropolitan city. Because it is difficult to reach, the oral health (OH) status of these residents is unknown. The objectives of this study are (1) to design a tool and strategy to collect OH information in a low-income community, (2) to characterize the OH status and related factors among low-income adults, and (3) to identify the explanatory factors for their OH status. MATERIALS AND METHODS: Mobile screening clinics were established in the gathering centers of the DTES, and those of 19 years of age or older were recruited. Data were collected through survey interviews and clinical examinations. Potential explanatory factors were investigated by regression analysis. RESULTS: The 356 screened participants were mostly males, middle-aged, less educated, and living with low income (≤CAD$20,000/y). About 80% had dental coverage, mostly from public programs (94%). Many (86%) perceived a dental need. Among dentate participants (n = 306), on average, 3.8 decayed, 8.6 missing, 4.9 filled teeth, and a care index of 41.5% were observed. Social factors (barriers to care and length of DTES residence), dental hygiene (brushing/flossing), and personal (hepatitis C virus infection/methadone usage) factors contributed to their care index level. CONCLUSION: This is the first time that comprehensive information regarding OH status has been collected from a low-income, inner-city community in Canada. Further investigations in the challenges and needs in accessing dental care may develop solutions for better OH in similar communities.

An actionable test using loss of heterozygosity in identifying high-risk oral premalignant lesions.

OBJECTIVES: To develop an actionable test using fluorescence capillary electrophoresis (FCE) to assess loss of heterozygosity (LOH) of histologically similar low-grade lesions (LGLs) to identify high-risk lesions for oral cancer progression. STUDY DESIGN: To determine the cutoffs of LOH, the FCE results of 52 surgical margin samples were used to compare with the existing LOH results from the previously validated 32 P-GE approach. Using the developed FCE workflow, an independent set of 102 LGLs with known progression status was used to determine the LOH molecular risk (MR) patterns and associated risk of progression. RESULTS: Using 65% cutoff LOH-FCE, the agreement of LOH-32 P-GE had an average of 82.3% (76.8-87.8). Compared with nonprogressors (n = 61), anatomic site and MR patterns (LOH at 9 p21, 3 p14, or 17 p13) were independent risk factors. High-risk profile of tongue and MR3 (LOH at 9 p21 and/or 3 p14 and 17 p13) was significantly associated with progression (hazard ratio [HR] 6.7; 95% confidence interval [CI] 2.6-17.6) with specificity of 98.4% at identifying progressors. CONCLUSIONS: We have developed an objective test using LOH to stratify the risk of LGLs. With further validation, it can be used in the clinical settings to provide clinicians additional information guiding the management of these lesions.

Decision making on detection and triage of oral mucosa lesions in community dental practices: screening decisions and referral.

UNLABELLED: Oral cancer is a substantial, often unrecognized issue globally, with close to 300 000 new cases reported annually. It is a management conundrum: a cancer site that is easily examined; yet more than 40% of oral cancers are diagnosed at a late stage when prognosis is poor and treatment can be devastating. Opportunistic screening within the dental office could lead to earlier diagnosis and intervention with improved survival. OBJECTIVE: To describe how clinicians make decisions about referral based on the risk classification of the lesion. METHODS: Eighteen dentists from 15 dental offices participated in a 1-day workshop on oral cancer screening. Participants then screened patients (medical history, conventional oral exam, fluorescent visualization examination) in-office for 11 months, triaging patients by apparent clinical risk: low risk (common benign conditions, geographic tongue, candidiasis, trauma), intermediate risk (lichenoid lesions) and high risk (white or red lesions or ulcers without apparent cause). Clinicians made the decision on which lesions to reassess in 3 weeks based on risk assessment and clinical judgment. Lesions of concern were seen by a community facilitator or referred to an oral medicine specialist. RESULTS: Of 2542 patients were screened, and 389 lesions were identified (15% of patients). 350 were determined to be low risk (90%), 19 intermediate risk (IR) (5%), and 20 high risk (HR) (5%). One hundred and sixty-six (43%) patients were recalled for 3-week reassessment: 90% of HR lesions, 63% of IR lesions (63%), and 39% of low-risk lesions. Compliance to recall was high (92% of cases). Reassessment eliminated the referral of 99/166 (60%) of lesions that had resolved. six lesions were biopsied with three low-grade dysplasias identified. CONCLUSIONS: Three key decision points were tested: risk assessment, need for reassessment, and need for referral. A 3-week reassessment appointment was invaluable to prevent the unnecessary referral due to confounders. There is a need for a well-defined triage pathway to facilitate oral cancer screening and a methodical and consistent approach to opportunistic screening in the dental office.

Influence of fluorescence on screening decisions for oral mucosal lesions in community dental practices.

BACKGROUND: Quality of oral screening examinations is dependent upon the experience of the clinician and can vary widely. Deciding when a patient needs to be referred is a critical and difficult decision for general practice clinicians. A device to aid in this decision would be beneficial. The objective of this study was to to examine the utility of direct fluorescence visualization (FV) by dental practitioners as an aid in decision-making during screening for cancer and other oral lesions. METHODS: Dentists were trained to use a stepwise protocol for evaluation of the oral mucosa: medical history, head, neck and oral exam, and fluorescent visualization exam. They were asked to use clinical features to categorize lesions as low (LR), intermediate (IR), or high (HR) risk and then to determine FV status of these lesions. Clinicians made the decision of which lesions to reassess in 3 weeks and based on this reassessment, to refer forward. RESULTS: Of 2404 patients screened over 11 months, 357 initially had lesions with 325 (15%) identified as LR, 16 (4.5%) IR, and 16 (4.5%) HR. Lesions assessed initially as IR and HR had a 2.7-fold increased risk of FV loss persisting to the reassessment appointment versus the LR lesions. The most predictive model for lesion persistence included both FV status and lesion risk assessment. CONCLUSION: A protocol for screening (assess risk, reassess, and refer) is recommended for the screening of abnormal intraoral lesions. Integrating FV into a process of assessing and reassessing lesions significantly improved this model.

The use of quantitative cytology in identifying high-risk oral lesions in community practice.

OBJECTIVE: This study evaluated whether quantitative cytology (QC) can disclose abnormal DNA content (aneuploidy) and abnormal nuclear morphology of high-risk potentially malignant disorders (PMDs) of the oral mucosa found in the community in reference to clinicohistopathologic features. STUDY DESIGN: A total of 171 patients at community-based clinic with suspicious oral lesions were evaluated with concurrent but independent histopathologic and QC assessments. RESULTS: QC-positive results were associated with oral lesions with higher clinical risk factors: large size, nonhomogeneous surface texture, and located at high-risk anatomic sites. Only 3% of benign/reactive and 5% of low-risk PMDs were QC positive, while 92% of high-risk PMDs and 88% of squamous cell carcinomas (SCCs) were QC positive. The sensitivity and specificity of QC for detection of high-grade dysplasia/SCC were 89% and 97%. CONCLUSIONS: QC could serve as an adjunctive tool for the detection of high-risk PMD/SCC requiring immediate clinical care.

Management of Frey syndrome using botulinum neurotoxin: a case report.

Frey syndrome is manifested clinically by hemifacial flushing and sweating after a gustatory stimulus. Frey syndrome is usually secondary to traumatic injury in the parotid region and is thought to be the result of misdirected re-sprouting of damaged autonomic nerve fibres. In this case report, we highlight the clinical and psychosocial aspects of Frey syndrome from a patient"s perspective, outline the pathophysiology of the condition and current management strategies, and describe the use of botulinum neurotoxin in the treatment of Frey syndrome.

Evaluation of a suspicious oral mucosal lesion.

Dentists who encounter a change in the oral mucosa of a patient must decide whether the abnormality requires further investigation. In this paper, we describe a systematic approach to the assessment of oral mucosal conditions that are thought likely to be premalignant or an early cancer. These steps, which include a comprehensive history, step-by-step clinical examination (including use of adjunctive visual tools), diagnostic testing and formulation of diagnosis, are routinely used in clinics affiliated with the British Columbia Oral Cancer Prevention Program (BC OCPP) and are recommended for consideration by dentists for use in daily practice.

Biopsy and histopathologic diagnosis of oral premalignant and malignant lesions.

Accurate diagnosis of premalignant or malignant oral lesions depends on the quality of the biopsy, adequate clinical information and correct interpretation of the biopsy results. The purpose of this paper is to review the procedures for obtaining appropriate biopsy samples, and the criteria for diagnosing and grading dysplasias. The World Health Organization's description of the architectural and cytologic epithelial changes that characterize dysplasia is detailed, and guidelines for following up patients with premalignant and malignant lesions are provided. The benefits of using the centralized services and expertise of the British Columbia Oral Biopsy Service are also reviewed.

Direct fluorescence visualization of clinically occult high-risk oral premalignant disease using a simple hand-held device.

BACKGROUND: A considerable proportion of oral cancer and precancer is not clinically apparent and could contribute significantly to the late diagnosis and high mortality of oral cancer. A simple method to identify such occult change is needed. METHODS: Patients in the Oral Dysplasia Clinics at British Columbia are currently being examined with a simple hand-held device that permits the direct visualization of alterations to autofluorescence in the oral cavity. Tissue showing loss of autofluorescence is biopsied. RESULTS: We present 3 representative cases in which occult lesions were identified with fluorescence visualization during longitudinal follow-up, resulting in the diagnosis of a primary dysplasia in case 1, a second primary cancer in case 2, and cancer recurrence in case 3. CONCLUSIONS: This is the first report of the diagnosis of occult oral disease using a simple noninvasive device. These early examples indicate the potential value of this technology to guide the management of patients with oral lesions, facilitating the detection of high-risk changes not apparent with white-light visualization.

Fluorescence visualization detection of field alterations in tumor margins of oral cancer patients.

PURPOSE: Genetically altered cells could become widespread across the epithelium of patients with oral cancer, often in clinically and histologically normal tissue, and contribute to recurrent disease. Molecular approaches have begun to yield information on cancer/risk fields; tissue optics could further extend our understanding of alteration to phenotype as a result of molecular change. EXPERIMENTAL DESIGN: We used a simple hand-held device in the operating room to directly visualize subclinical field changes around oral cancers, documenting alteration to fluorescence. A total of 122 oral mucosa biopsies were obtained from 20 surgical specimens with each biopsy being assessed for location, fluorescence visualization (FV) status, histology, and loss of heterozygosity (LOH; 10 markers on three regions: 3p14, 9p21, and 17p13). RESULTS: All tumors showed FV loss (FVL). For 19 of the 20 tumors, the loss extended in at least one direction beyond the clinically visible tumor, with the extension varying from 4 to 25 mm. Thirty-two of 36 FVL biopsies showed histologic change (including 7 squamous cell carcinoma/carcinomas in situ, 10 severe dysplasias, and 15 mild/moderate dysplasias) compared with 1 of the 66 FV retained (FVR) biopsies. Molecular analysis on margins with low-grade or no dysplasia showed a significant association of LOH in FVL biopsies, with LOH at 3p and/or 9p (previously associated with local tumor recurrence) present in 12 of 19 FVL biopsies compared with 3 of 13 FVR biopsies (P=0.04). CONCLUSIONS: These data have, for the first time, shown that direct FV can identify subclinical high-risk fields with cancerous and precancerous changes in the operating room setting.

Simple device for the direct visualization of oral-cavity tissue fluorescence.

Early identification of high-risk disease could greatly reduce both mortality and morbidity due to oral cancer. We describe a simple handheld device that facilitates the direct visualization of oral-cavity fluorescence for the detection of high-risk precancerous and early cancerous lesions. Blue excitation light (400 to 460 nm) is employed to excite green-red fluorescence from fluorophores in the oral tissues. Tissue fluorescence is viewed directly along an optical axis collinear with the axis of excitation to reduce inter- and intraoperator variability. This robust, field-of-view device enables the direct visualization of fluorescence in the context of surrounding normal tissue. Results from a pilot study of 44 patients are presented. Using histology as the gold standard, the device achieves a sensitivity of 98% and specificity of 100% when discriminating normal mucosa from severe dysplasia/carcinoma in situ (CIS) or invasive carcinoma. We envisage this device as a suitable adjunct for oral cancer screening, biopsy guidance, and margin delineation.