Loss Of Autofluorescence Research Articles

The onset and progression of oral potentially malignant disorders in Fanconi anemia patients: Highlighting early detection of oral cancer

In 2020, Fanconi anemia (FA) was classified as a syndrome with insufficient epidemiological evidence in the oral potentially malignant disorder (OPMD) group by the WHO Collaborating Centre. The paucity of case reports on FA-associated OPMD limits evidence-based management, and such cases have not been analyzed collectively in detail. Hence, the objective of this short communication is to summarize the evidence on the onset and progression of OPMD in FA patients, so as to better understand the natural history of oral cancer development in patients affected by FA. A total of 11 eligible papers containing 1332 FA patients are involved in onset and progression of OPMD in FA patients. Of these, 186 (14.0%) were diagnosed with OPMD. With available data from 4 follow-up studies, 30 (41.1%) of 73 FA patients compatible with OPMD further developed into OSCC at young age (10–30 years old). The evidence on FA with malignant potential comprise clinical epidemiology, oral cytology abnormalities, DNA aneuploidy, loss of autofluorescence, loss of heterozygosity, high-risk human papillomavirus infection, DNA mutations in saliva and plasma samples. Collectively, these can consummate the evidence on FA as a syndrome that may potentiate cancer development in oral cavity mentioned by the WHO. Importantly, it highlights close surveillance is instrumental for FA patients with OPMD to early detect oral cancer.

COMPLEMENT INHIBITION FOR GEOGRAPHIC ATROPHY: Review of Salient Functional Outcomes and Perspective.

To evaluate available rationale and outcomes of randomized trial results for complement inhibition for geographic atrophy. Data from recently completed randomized trials of complement inhibition, particularly for pegcetacoplan and avacincaptad pegol, were evaluated for both the outcome, area of autofluorescence loss, and functional vision tests. Pegcetacoplan 2 mg showed statistically significant reduction in expansion of the area of autofluorescence loss with monthly, but not every-other-month dosing, in a 12-month phase two trial. Nearly 40% of patients recruited for the monthly arm did not complete the treatment. In two parallel phase 3 studies there was a statistically significant reduction in the area of atrophy in one but not both studies as compared with untreated controls. Data released at 24 months follow-up showed statistically significant reduction in the area of autofluorescence-detected atrophy in both studies compared with sham. Patients did not show functional difference in best-corrected visual acuity, maximum reading speed, Functional Reading Independence Index, and mean microperimetry threshold sensitivities in the treatment versus sham arms. Avacincaptad pegol was evaluated in two randomized pivotal studies and showed a statistically significant reduction in the expansion of autofluorescence loss at 12 months. Patients in the treatment arms did not show any difference as compared with sham in the best-corrected visual acuity or low luminance visual acuity, the only functional outcomes mentioned. Both drugs increased the risk of macular neovascularization. Both avacincaptad pegol and pegcetacoplan show significant differences compared with sham in autofluorescence imaging but no benefit in visual function at 12 and 24 months, respectively.

Assessment of DNA Ploidy in Oral Potentially Malignant Disorders Using a VELscope

Introduction: The device VELscope (Visually enhanced lesion scope) is intended as an aid to the clinician to complement a conventional white light exam (whether it be a general oral cavity examination or examination of a particular lesion) to detect abnormal tissue that might have otherwise been overlooked. In this study, it was used concomitantly with ploidy status to evaluate the malignancy potential of Oral Potentially Malignant Disorders (OPMD). Aim: To evaluate potential of Deoxyribonucleic Acid (DNA) ploidy as a preliminary adjunct to oral biopsy to identify patients with aneuploidy for the necessity of biopsy, and to assess the sensitivity of VELscope in the determination of accurate clinical parameters of the lesions in the oral mucosa. Materials and Methods: This in-vivo cross-sectional study was carried out in Department of Oral and Maxillofacial Pathology and Oral Microbiology, Bharati Vidyapeeth Deemed to be University, Pune from February 2020 to February 2021. Clinically diagnosed OPMD were included in the study, after verification of clinical extension boundaries by VELscope. DNA ploidy status was evaluated with DNA image cytometry and exclusive individuals with aneuploidy were biopsied followed by histopathological evaluation and surgical removal of the lesion. One way Analysis of Variance (ANOVA) test was used to compare the mean in 4 study groups [Oral leukoplakia, Oral submucous fibrosis, Oral lichen planus and Tobacco pouch keratosis] vs control. Sensitivity, specificity, positive predictive value and negative positive value was calculated by using the ROC (Receiver Operating Characteristic Curve) analysis. A p-value <0.05 was considered as significant for evaluating sensitivity of VELscope and evaluation of ploidy status. Results: The study included patients whose mean age was 45.8 years. The groups had tobacco habits in one of the various forms [smoke or smokeless] both in experimental and control group. VELscope could identify the clinical borders of the OPMDs considered via the loss of autofluorescence. Lesion borders were identified precisely with the loss of autofluorescence. It was quite helpful especially in aneuploid cases where the whole margin could be removed leaving no genetically aberrant cells behind. Oral submucous fibrosis was concluded to have maximal sensitivity of 86.7%, 100% in specificity and positive predictive and 88.2% negative predictive value. Conclusion: DNA ploidy could decipher the malignancy potential and could identify the individuals who needed biopsy. VELscope was able to mark the clinical diameters which were not visible to naked eyes clinically. Oral submucous fibrosis was found to be group with the maximum potential of malignancy followed by Oral Lichen Planus, Oral Leukoplakia and Tobacco Pouch Keratosis.

Mildly dysplastic oral lesions with optically-detectable abnormalities share genetic similarities with severely dysplastic lesions

ObjectiveOptical imaging studies of oral premalignant lesions have shown that optical markers, including loss of autofluorescence and altered morphology of epithelial cell nuclei, are predictive of high-grade pathology. While these optical markers are consistently positive in lesions with moderate/severe dysplasia or cancer, they are positive only in a subset of lesions with mild dysplasia. This study compared the gene expression profiles of lesions with mild dysplasia (stratified by optical marker status) to lesions with severe dysplasia and without dysplasia. Materials and methodsForty oral lesions imaged in patients undergoing oral surgery were analyzed: nine without dysplasia, nine with severe dysplasia, and 22 with mild dysplasia. Samples were submitted for high throughput gene expression analysis. ResultsThe analysis revealed 116 genes differentially expressed among sites without dysplasia and sites with severe dysplasia; 50 were correlated with an optical marker quantifying altered nuclear morphology. Ten of 11 sites with mild dysplasia and positive optical markers (91%) had gene expression similar to sites with severe dysplasia. Nine of 11 sites with mild dysplasia and negative optical markers (82%) had similar gene expression as sites without dysplasia. ConclusionThis study suggests that optical imaging may help identify patients with mild dysplasia who require more intensive clinical follow-up. If validated, this would represent a significant advance in patient care for patients with oral premalignant lesions.

USEFULNESS OF FLUORESCENCE IMAGING FOR EARLY DIAGNOSIS OF ORAL POTENTIALLY MALIGNANT DISORDERS

Objectives The objective of this study was to review in the literature the use of fluorescence imaging as a method of early diagnosis in patients with oral potentially malignant disorders. Study Design A bibliographic survey covering the previous 10 years was carried out in the PubMed, MEDLINE, and LILACS databases. Results Thirty articles were collected, and all were selected. Most studies have stated that fluorescence is helpful to complement the clinical examination performed under conventional white light, improving the accuracy of the lesion margin and exposing the loss of autofluorescence, meaning potential oral epithelial dysplasia (OED) or malignancy. As a complement, this method has been described in 4 studies, showing a significant improvement in the diagnostic results and a slight increase in the effectiveness of the clinical examination when performed by a specialist. However, 5 articles stated the need for studies in this area to confirm the accuracy of fluorescence. It remains undefined in the detection of erythematous lesions of mild inflammation because it can result in a false-positive test. Conclusions Fluorescence imaging is helpful as a diagnostic aid, with biopsy being the reference technique for assessing OED. The objective of this study was to review in the literature the use of fluorescence imaging as a method of early diagnosis in patients with oral potentially malignant disorders. A bibliographic survey covering the previous 10 years was carried out in the PubMed, MEDLINE, and LILACS databases. Thirty articles were collected, and all were selected. Most studies have stated that fluorescence is helpful to complement the clinical examination performed under conventional white light, improving the accuracy of the lesion margin and exposing the loss of autofluorescence, meaning potential oral epithelial dysplasia (OED) or malignancy. As a complement, this method has been described in 4 studies, showing a significant improvement in the diagnostic results and a slight increase in the effectiveness of the clinical examination when performed by a specialist. However, 5 articles stated the need for studies in this area to confirm the accuracy of fluorescence. It remains undefined in the detection of erythematous lesions of mild inflammation because it can result in a false-positive test. Fluorescence imaging is helpful as a diagnostic aid, with biopsy being the reference technique for assessing OED.

Autofluorescence imaging as a noninvasive tool of risk stratification for malignant transformation of oral leukoplakia: A follow-up cohort study

BackgroundThere is few longitudinal study on oral potentially malignant disorder (OPMDs) focusing on oral leukoplakia (OLK) with clinical endpoints to investigate the role of autofluorescence imaging (AFI) for surveilling the malignant transformation. MethodsBased on our previous prospective diagnostic study enrolled 517 OPMD patients, 184 OLK patients were retrieved to further investigate the role of AFI using VELscope in predicting malignant transformation. During a median follow-up period of 44 months, 19 (10.33%) developed into oral cancer. ResultsOLK patients were divided into loss of autofluorescence (LAF, n = 124) and retention of autofluorescence (RAF, n = 60) groups according to the results of AFI. Interestingly, difference between malignant transformation rate (MTR, 14.52%) of group LAF and overall MTR (10.33%) was not significant, but MTR (1.67%) of group RAF was significantly lower than overall MTR. Kaplan-Meier and Cox-regression analyses revealed that LAF could not directly distinguish the high-risk lesions, but RAF significantly discriminate the low-risk lesions. Importantly, time-dependent ROC curve analysis found that the sensitivity and negative predictive value (NPV) of AFI for the prediction of malignant transformation was 100% and 100% (2-year follow-up), 94.7% and 98.3% (5-year follow-up), respectively. Also, calibration curve and decision curve analyses showed high levels of predictive value and clinical relevance. ConclusionThis follow-up cohort study firstly evaluated AFI using VELscope for risk stratification of OLK malignant transformation. Whether conservative management is appropriate for OLK patients with RAF imaging due to minimal rate and risk of malignant transformation and great specificity and NPV is required to be further investigated.

Velscope guided oral cancer screening

Context:Early oral cancer diagnosis is an important factor in reducing cancer-associated mortality and morbidity. Traditional oral cancer screening by Conventional Oral Examination (COE) is limited. A new approach in this regard is the use of “optical biopsy systems” like VELscope.Aims:Hence, this study aimed to determine the diagnostic value of VELscope, an autofluorescence-based handheld device in detection of dysplasia and oral squamous cell carcinoma (OSCC) in general oral potentially malignant disorder.Settings and Design:A prospective, cross-sectional study was conducted at a tertiary hospital in western Maharashtra, India.Materials and Methods:Thirty patients with presence of clinically suspicious oral lesions were included, and for each lesion, both the COE and Autofluorescence Examination by VELscope were carried out at the same appointment by different experts. All lesions were biopsied and histopathological findings were documented and analyzed.Statistical Analysis Used:Sensitivity, specificity, positive predictive value and negative predictive value were estimated to determine the accuracy of VELscope examination analysis outcome.Results:The study included a total of 30 subjects including 19 (63.33%) males and 11 (26.66%) females. Autofluorescence examination by VELscope (AFV) revealed Autofluorescence Loss in 24 (80%) patients, while 6 (20%) patients showed Autofluorescence Retained. Comparison between the “high-risk lesions” (moderate/severe dysplasia and OSCC) and “low-risk lesions” (no/mild dysplasia) showed a 90.47% sensitivity but only 44.44% specificity.Conclusions:The findings of our study indicate that although AFV cannot be a substitute for COE, it can be used as a potential complementary diagnostic aid in surveillance of the high-risk patient population.

Non-Invasive Diagnostic System Based on Light for Detecting Early-Stage Oral Cancer and High-Risk Precancerous Lesions-Potential for Dentistry.

Simple SummaryThe early detection of oral cancer and oral potentially malignant disorders can facilitate minimum intervention and subsequent improvements in prognosis and quality of life after treatment. Recently, several non-invasive adjunctive fluorescence-based detection systems have improved the accuracy of detection and diagnosis of oral cancer and oral potentially malignant disorders. This article summarizes current knowledge about fluorescence-based diagnostic methods and discusses their benefits and drawbacks from a clinical viewpoint.Oral health promotion and examinations have contributed to the early detection of oral cancer and oral potentially malignant disorders, leading to the adaptation of minimally invasive therapies and subsequent improvements in the prognosis/maintenance of the quality of life after treatments. However, the accurate detection of early-stage oral cancer and oral epithelial dysplasia is particularly difficult for conventional oral examinations because these lesions sometimes resemble benign lesions or healthy oral mucosa tissues. Although oral biopsy has been considered the gold standard for accurate diagnosis, it is deemed invasive for patients. For this reason, most clinicians are looking forward to the development of non-invasive diagnostic technologies to detect and distinguish between cancerous and benign lesions. To date, several non-invasive adjunctive fluorescence-based detection systems have improved the accuracy of the detection and diagnosis of oral mucosal lesions. Autofluorescence-based systems can detect lesions as a loss of autofluorescence through irradiation with blue-violet lights. Photodynamic diagnosis using 5-aminolevulinic acid (ALA-PDD) shows the presence of very early oral cancers and oral epithelial dysplasia as a red fluorescent area. In this article, currently used fluorescence-based diagnostic methods are introduced and discussed from a clinical point of view.

Clinico-pathological correlation of optical fluorescence imaging in oral mucosal lesions.

This study aimed to identify clinical and pathological characteristics of oral mucosal lesions that may be predictive of optical autofluorescence imaging patterns. Clinical data and archival histopathological material were collected from patients who presented with at least one oral mucosal lesion and underwent assessment via conventional oral examination, optical autofluorescence imaging and histopathological analysis. An open-source digital pathology image analysis software was used to perform histomorphometric measurements. Classification and regression trees were used to determine histopathological characteristics most predictive of a clinical autofluorescence outcome. Histomorphometric features associated with tissue architecture, epithelial changes, inflammation and vasculature were found to be significantly associated with autofluorescence patterns. Diascopic fluorescence was found to be significantly predicted by lichenoid inflammation and was significantly associated with a diagnosis of oral lichen planus. Loss of autofluorescence with partial blanching was significantly associated with histopathological features noted in dysplastic and malignant lesions. This study provides evidence for the use of diascopic fluorescence as a technique to aid in clinical differentiation of benign inflammatory lesions from potentially malignant pathology. Based on the findings of this study, optical fluorescence imaging is a technique of added value in discernment of oral mucosal lesions, and our results support its clinical use.

Interobserver and intraobserver variability affecting the assessment of loss of autofluorescence of oral mucosal lesions.

The assessment of loss of tissue autofluorescence (LAF) has been proposed as an adjunct to comprehensive oral examination to enhance the detection of mucosal lesions harbouring dysplasia or carcinoma. The assessment of LAF is not based on completely objectified parameters therefore intraobserver and interobserver variability cannot be neglected alongside the issue of correct interpretation of LAF. The present study evaluated intraobserver and interobserver variability in the clinical assessment of LAF as performed by oral medicine practitioners (OMPs) or general dental practitioners (GDPs). Couples of clinical pictures, acquired under white incandescent dental operatory light and during the assessment of LAF performed by VELscope were retrieved. Four OMPs and eight GDPs were asked to assess the pictures and to score the LAF. Kappa statistics allowed the assessment of intra- and inter-observer related variability. Pictures of 109 lesions representative of all oral mucosal sites and clinical appearances were selected. OMPs had a better intraobserver agreement than GDPs (substantial versus moderate). The moderate (k = 0.506) interobserver agreement observed among both OMPs and GDPs in a 2-score model (positive versus negative), lowered down to poor values only among GDPs when a 3-score or 4-score model (including uncertain judgements) was applied. A good agreement (k>0.8) was never observed and the present results are similar to previously reported data about conventional oral examination. Irrespective of the diagnostic accuracy, the assessment of AF seems not to be able to improve observer-related variability in the clinical assessment of oral mucosal lesions.

Potential role of autofluorescence imaging in determining biopsy of oral potentially malignant disorders: A large prospective diagnostic study

Autofluorescence examination of oral tissues using the VELscope has been suggested as an adjunctive tool for cancer detection and diagnosis. This study aimed to determine the diagnostic value of VELscope in a large prospective study of 517 patients with oral potentially malignant disorders (OPMD). For the outcome assessments of discrimination of carcinoma form general OPMD and distinguishing high-risk lesions (moderate/severe dysplasia and carcinoma) from low-risk lesions (no/mild dysplasia), high sensitivity (100% and 95.9% respectively) and negative predictive value (100% and 98.2% respectively) were observed. All the carcinoma and showed loss of autofluorescence (LAF) and only 3 (0.6%) moderate/severe dysplasia were observed without LAF.These data indicate that the cases without LAF using VELscope substantially rule out the presence of high-risk lesions including cancer. This may prove to be useful specially to alleviate patient anxiety regarding a clinically suspicious oral lesion without the LAF, and to avoid a unnecessary biopsy for these cases. Collectively, a perspective to highlight was that a no biopsy strategy may be appropriate for OPMD without LAF using VELscope after conventional oral examination.

Usefulness of optical fluorescence imaging in identification and triaging of oral potentially malignant disorders: A study of VELscope in the LESIONS programme.

Optical fluorescence imaging (OFI) has been shown to enhance the detection of oral potentially malignant disorders (OPMD), by providing additional information about the nature of these lesions, with loss of autofluorescence (LAF) signifying potential underlying oral epithelial dysplasia (OED) or malignancy. This study aims to assess the usefulness of OFI in identification and triaging of OPMDs by primary oral health practitioners, in a community setting. Participants were recruited from the LESIONS programme in South-East Queensland, Australia. Participants underwent both conventional oral examination (COE) of mucosal tissues as well as screening with OFI by trained dentists and oral health therapists (OHTs). Lesions were classified as homogeneous, non-homogeneous, lichenoid or other. The study protocol included blanching of lesions detected with VELscope® (diascopy) and a two-week review and reassessment of all lesions to reduce false-positive findings. Analysis of decisions made by dentists and OHTs about lesion identification and triaging was undertaken. A total of 360 participants underwent both COE and OFI screening and presented with at least one lesion detected with either method. Lesion size increased and border distinctness and visibility improved with OFI. Of the 170 lesions that underwent two-week reassessment, 70% resolved or were confirmed as benign. OHTs were more likely to request review and referral for OPMDs compared to dentists. Optical fluorescence imaging is a useful tool to help dentists and OHTs identify and triage patients with possible OPMD in the primary care setting.

ORAL TISSUE CONDITION IN PROSTHODONTIC GERIATRIC PATIENTS

The world population is continuously getting older in developed and undeveloped countries. According to WHO over 20% of the population in Europe is over 65 years old. Geriatric population in North Macedonia is presented with 16% and with high prevalence of anodontia over 45, 1%, poor oral hygiene and urgent demand of prosthodontics treatment. Aging is a process with different and specific changes of all systems and organs, including the orophacial system. In this population, the number of remaining teeth is evidently reduced, their condition has changed, and soft and hard oral tissues are affected. It is very important to discover all the factors that lead to the development of leukoplakia and other precancerous lesions as early as possible. Precancerous lesions and oral carcinoma are more common in the elderly compared to the younger population. Almost 90% of oral cancers occur in patients older than 50 years. Early detection of oral mucosa diseases in elderly and potentially malignant disorders requires careful examination because of high malignant transformation frequency. The aim of the study was to determine the condition of oral soft tissues in geriatric prosthodontics patients, their general health and the impact of oral health on their quality of life. The value of the VELscope instrument as an adjunct to oral examination in geriatric prosthodontics patients was also evaluated. The study was performed in the University Dental Clinic Ss’ Panteleimon in Skopje, NM. Clinical protocol for 300 patients over 60 years old was questionnaire with risk factors, clinical intra and extraoral observation and VELscope mucosal tissue examination. Tissue changes were classified as inflammatory, traumatic, dysplastic and other. Abnormal tissue was associated with auto fluorescence loss and dark appearance in contrast to the surrounding healthy tissue. In 21% of patients abnormal premalignant lesions were detected. Leukoplakia was the most common premalignant disorder 42 (12, 6%), 16 (4, 8%) had lichen planus, 6 (1, 8%) cheilitis actinica and 5 patients (1, 5%) were diagnosed with squamous cell carcinoma. We found 118 inflammatory lesions (35, 4%) where 58 (17, 4%) were denture stomatitis, 18 (5, 4%) angular cheilitis, traumatic lesions were found in 38 patients (11, 4%) and 4 (1, 2%) were diagnosed with epulis fissuratum. In 54 patients (16, 2%) we diagnosed infectious changes from which aphtous ulcerations and candidiasis were most common. The VELscope fluorescence instrument can be used as a part of diagnostic process to detect abnormal tissue and oral lesions that might have been overlooked. A validation of the study was carried out and it was approved by the ethics committee of the Faculty for dentistry Ss Cyril and Methodius University Skopje on 18.12.2018. No. 02-38363.

Auto-Fluorescence and Histopathologic Evaluation of Medication-Related Osteonecrosis of the Jaws: Perspectives for Treatment.

Auto-fluorescence (AF) of healthy bone tissue has recently been described. Loss of AF (LAF) has, on the contrary, been reported in necrotic bone. Further, the use of LAF as a possible guidance to distinguish viable from necrotic bone during surgical treatment of osteonecrosis has been proposed. The aim of this study is to detail 8 patients of medication-related osteonecrosis of the jaws treated through an AF-guided surgical resection. The authors also provide the histopathologic description of hypo-fluorescent and hyper-fluorescent bone in each patient. After removal of necrotic bone block, Er:YAG laser was used for vaporizing further necrotic bone, up to the detection of strongly hyper-fluorescent bone. Samples of hyper-fluorescent bone were collected around areas of necrosis. Histopathologic evaluation revealed viable bone tissue in all hyper-fluorescent specimens. On the basis of these data, AF-guided surgical resection could be effective in highlighting surgical margins of necrotic bone tissue and it might have some utility in a range of applications of bone surgery.

Exophytic verrucous hyperplasia in oral submucous fibrosis: A single-center study.

Introduction:The present study analyzed the occurrence of exophytic verrucous hyperplasia (EVH) in the background of oral submucous fibrosis (OSF), which presents clinically as a solitary verrucopapillary lesion (VPL) mimicking malignancy. We also aimed to obtain additional information on VELscope appearance and histopathological features of EVH.Materials and Methods:The prevalence of EVH in OSF background was assessed from January 2014 to December 2018 using VELscope and histopathological examination.Results:Six hundred and sixty-two OSF patients were examined. Thirteen patients presented with solitary VPL in OSF background. A VELscope examination found ten cases with increased autofluorescence (fluorescence visualization increase, FVI), two cases with autofluorescence loss (fluorescence visualization loss, FVL), whereas one case exhibited dual autofluorescence (focal areas of FVL within FVI regions). Histopathologic examination revealed two FVL cases as oral verrucous carcinoma (OVC) and oral squamous cell carcinoma (OSCC) and one dual autofluorescence case as OVC, while six FVI cases showed nondysplastic epithelium having verrucopapillary pattern without connective tissue invasion, consistent with the clinicopathological diagnosis of EVH.Conclusion:The present study demonstrated the evidence of EVH in OSF background, which on histopathological examination revealed nondysplastic epithelium exhibiting the verrucopapillary pattern. A VELscope examination of these lesions showed increased autofluorescence, suggesting its nonneoplastic nature of clinically malignant-looking exophytic VPLs in OSF background. Present study suggests newer perspective for using the term oral verrucous hyperplasia (OVH) and EVH with justification and also proposes to introduce new terminology such as oral verrucous dysplasia and exophytic verrucous dysplasia.

Risk Stratification of Oral Potentially Malignant Disorders in Fanconi Anemia Patients Using Autofluorescence Imaging and Cytology-On-A Chip Assay

Fanconi anemia (FA) is a hereditary genomic instability disorder with a predisposition to leukemia and oral squamous cell carcinomas (OSCCs). Hematopoietic stem cell transplantation (HSCT) facilitates cure of bone marrow failure and leukemia and thus extends life expectancy in FA patients; however, survival of hematologic malignancies increases the risk of OSCC in these patients. We developed a “cytology-on-a-chip” (COC)–based brush biopsy assay for monitoring patients with oral potentially malignant disorders (OPMDs). Using this COC assay, we measured and correlated the cellular morphometry and Minichromosome Maintenance Complex Component 2 (MCM2) expression levels in brush biopsy samples of FA patients’ OPMD with clinical risk indicators such as loss of autofluorescence (LOF), HSCT status, and mutational profiles identified by next-generation sequencing. Statistically significant differences were found in several cytology measurements based on high-risk indicators such as LOF-positive and HSCT-positive status, including greater variation in cell area and chromatin distribution, higher MCM2 expression levels, and greater numbers of white blood cells and cells with enlarged nuclei. Higher OPMD risk scores were associated with differences in the frequency of nuclear aberrations and differed based on LOF and HSCT statuses. We identified mutation of FAT1 gene in five and NOTCH-2 and TP53 genes in two cases of FA patients’ OPMD. The high-risk OPMD of a non-FA patient harbored FAT1, CASP8, and TP63 mutations. Use of COC assay in combination with visualization of LOF holds promise for the early diagnosis of high-risk OPMD. These minimally invasive diagnostic tools are valuable for long-term surveillance of OSCC in FA patients and avoidance of unwarranted scalpel biopsies.

Pesticide dissipation and microbial community changes in a biopurification system: influence of the rhizosphere.

The dissipation of atrazine, chlorpyrifos and iprodione in a biopurification system and changes in the microbial and some biological parameters influenced by the rhizosphere of Lolium perenne were studied in a column system packed with an organic biomixture. Three column depths were analyzed for residual pesticides, peroxidase, fluorescein diacetate activity and microbial communities. Fungal colonization was analyzed by confocal laser scanning microscopy to assess the extent of its proliferation in wheat straw. The L. perenne rhizosphere enhanced pesticide dissipation and negligible pesticide residues were detected at 20-30cm column depth. Atrazine, chlorpyrifos and iprodione removal was 82, 89 and 74% respectively in the first 10cm depth for columns with vegetal cover. The presence of L. perenne in contaminated columns stimulated peroxidase activity in all three column depth sections. Fluorescein diacetate activity decreased over time in all column sections with the highest values in biomixtures with vegetal cover. Microbial communities, analyzed by PCR-DGGE, were not affected by the pesticide mixture application, presenting high values of similarity (>65%) with and without vegetal cover. Microbial abundance of Actinobacteria varied according to treatment and no clear link was observed. However, bacterial abundance increased over time and was similar with and without vegetal cover. On the other hand, fungal abundance decreased in all sections of columns after 40days, but an increase was observedin response to pesticide application. Fungal colonization and straw degradation during pesticide dissipation were verified by monitoring the lignin autofluorescence loss.

Characterization ofCDH3-Related Congenital Hypotrichosis With Juvenile Macular Dystrophy

Congenital hypotrichosis with juvenile macular dystrophy (HJMD) is a rare disorder presenting in childhood and adolescence with central visual disturbance and sparse scalp hair. Reported retinal imaging is lacking, and whether the condition is progressive remains unclear. To investigate a series of patients with HJMD due to biallelic mutations in CDH3 and thereby characterize the disorder. Ten patients from 10 families underwent detailed clinical assessment, including serial retinal imaging and electrophysiologic evaluation, at Moorfields Eye Hospital, St James's University Hospital, and Calderdale Royal Infirmary. Patients ranged in age from 3 to 17 years at onset and 5 to 57 years at last assessment. The molecular genetic investigation included bidirectional Sanger sequencing of all exons and intron-exon boundaries of CDH3 and whole-exome sequencing in 2 patients. The study was conducted from June 5, 2013, to January 15, 2016, with final follow-up completed on December 15, 2015. Results of clinical assessment and molecular genetic testing. All 10 patients (7 male and 3 female) presented with central visual disturbance in childhood and had lifelong sparse scalp hair with normal facial hair. Fundus examination revealed chorioretinal atrophy of the posterior pole contiguous with the disc in all but 1 patient that was associated with marked loss of autofluorescence on fundus autofluorescence imaging. Optical coherence tomography (OCT) demonstrated variable degrees of atrophy of the outer retina, retinal pigment epithelium, and choroid, with outer retinal tubulations frequently observed. One patient had mild disruption of the inner segment ellipsoid band on OCT and additional mild digit abnormalities. Electrophysiologic evaluation in 5 patients demonstrated macular dysfunction with additional mild, generalized retinal dysfunction in 2 patients. Eight patients had more than 1 evaluation; of these, 5 patients showed deterioration of visual acuity over time, 1 patient remained stable, and 2 patients had severe visual loss at presentation that precluded assessment of visual deterioration. The area of atrophy did not progress with time, but retinal thickness decreased on OCT. Electrophysiologic evaluation in 1 patient found deterioration of macular function after 13 years of follow-up, but the mild, generalized photoreceptor dysfunction remained stable. Biallelic mutations were identified in all patients, including 6 novel mutations. These results suggest that CDH3-related disease is characterized by a childhood-onset, progressive chorioretinal atrophy confined to the posterior pole. The disease is readily distinguished from other juvenile macular dystrophies by the universally thin and sparse scalp hair. Patients may have additional limb abnormalities.

Acridine Orange Indicates Early Oxidation of Wood Cell Walls by Fungi.

Colonization of wood blocks by brown and white rot fungi rapidly resulted in detectable wood oxidation, as shown by a reduced phloroglucinol response, a loss of autofluorescence, and acridine orange (AO) staining. This last approach is shown to provide a novel method for identifying wood oxidation. When lignin was mildly oxidized, the association between AO and lignin was reduced such that stained wood sections emitted less green light during fluorescence microscopy. This change was detectable after less than a week, an interval that past work has shown to be too short for significant delignification of wood. Although fungal hyphae were observed in only a few wood lumina, oxidation was widespread, appearing relatively uniform over regions several hundred micrometers from the hyphae. This observation suggests that both classes of fungi release low molecular weight mild oxidants during the first few days of colonization.

Autofluorescence imaging in recurrent oral squamous cell carcinoma.

The survival of patients with oral cancer is decreased by locoregional recurrence after an initial multimodal treatment. In order to identify lesions in the oral cavity for a possible recurrence, clinical evaluation as well as MRI or CT scanning is advised. The evaluation of mucosa lesions is hampered by changes related to radio- and chemotherapy as well as reconstruction with tissue flaps. Several techniques for easier identification of tissue abnormalities in the oral cavity have been advocated as adjuncts in order to facilitate identification. Especially methods using altered tissue fluorescence have gained much interest during the last decade. The aim of our prospective study was to evaluate fluorescence properties of undiagnosed mucosa lesions with the VELscope device in patients with multimodal treated oral cancer prior to histological confirmation. In total, 41 patients with a history of oral squamous cell carcinomas (OSCC) (19 females and 22 males) with undiagnosed mucosa lesions where included in the study. After clinical evaluation, examination and documentation using the VELscope® device were performed. Then, an incisional biopsy was performed. An autofluorescence loss indicating a malignant or dysplastic mucosa condition could be detected in six patients (14.6%); however, only one OSCC and one SIN revealed a complete autofluorescence loss. In four patients, OSCC was present in lesions with retained autofluorescence. Sensitivity and specificity for the VELscope® examination to identify malignant oral lesions by autofluorescence were 33.3 and 88.6%, respectively. The positive and negative predictive values were 33.3 and 88.6%, respectively. No statistical correlation between gender and lesion appearance versus autofluorescence loss could be detected. In contrast to mucosa lesions in patients with no prior treatment, the autofluorescence evaluation with the VELscope reveals no additional information in our analysis. Accordingly, invasive biopsies as gold standard are still needed to get sufficient evidence regarding potential malignancy in patients after multimodal treatment for oral cancer.