Short- and long-term effects of a community-based suicide prevention program: A Hong Kong experience.

BACKGROUND: A multidisciplinary, multilayer, community-based suicide prevention program (2008-2012) was implemented in the Eastern District, Hong Kong. This article documents the program and reports on short- and longer-term program evaluation. METHODS: Characteristics and rates of self-harm/suicidal behaviors and suicide deaths by age group and gender in the Eastern District before, during, and after the intervention were calculated and compared with the rest of Hong Kong, using Kruskal-Wallis and chi-squared tests, and Jonckheere-Terpstra and Cochran-Mantel-Haenszel tests for trend analyses. RESULTS: The program impacts varied by age and gender subgroups. Suicide rates in the Eastern District were lower compared to the rest of Hong Kong during the intervention period. They slowly rebounded after the intervention ceased; nevertheless, they remained lower than the rest of Hong Kong until 2016. The rates of self-harm continuously dropped and remained lower than the rest of Hong Kong. During the intervention period in the Eastern District, the age of people who died by suicide increased; more deaths occurred from jumping and fewer by charcoal burning. CONCLUSIONS: The program coincided with the lowered self-harm and suicide rates after the implementation. Some of the strategies need to be rebooted or routinely and continuously implemented to ensure the sustainability.

RAF1-MEK/ERK pathway-dependent ARL4C expression promotes ameloblastoma cell proliferation and osteoclast formation.

Ameloblastoma is an odontogenic neoplasm characterized by slow intraosseous growth with progressive jaw resorption. Recent reports have revealed that ameloblastoma harbours an oncogenic BRAFV600E mutation with mitogen-activated protein kinase (MAPK) pathway activation and described cases of ameloblastoma harbouring a BRAFV600E mutation in which patients were successfully treated with a BRAF inhibitor. Therefore, the MAPK pathway may be involved in the development of ameloblastoma; however, the precise mechanism by which it induces ameloblastoma is unclear. The expression of ADP-ribosylation factor (ARF)-like 4c (ARL4C), induced by a combination of the EGF-MAPK pathway and Wnt/ß-catenin signalling, has been shown to induce epithelial morphogenesis. It was also reported that the overexpression of ARL4C, due to alterations in the EGF/RAS-MAPK pathway and Wnt/ß-catenin signalling, promotes tumourigenesis. However, the roles of ARL4C in ameloblastoma are unknown. We investigated the involvement of ARL4C in the development of ameloblastoma. In immunohistochemical analyses of tissue specimens obtained from 38 ameloblastoma patients, ARL4C was hardly detected in non-tumour regions but tumours frequently showed strong expression of ARL4C, along with the expression of both BRAFV600E and RAF1 (also known as C-RAF). Loss-of-function experiments using inhibitors or siRNAs revealed that ARL4C elevation depended on the RAF1-MEK/ERK pathway in ameloblastoma cells. It was also shown that the RAF1-ARL4C and BRAFV600E-MEK/ERK pathways promoted cell proliferation independently. ARL4C-depleted tumour cells (generated by knockdown or knockout) exhibited decreased proliferation and migration capabilities. Finally, when ameloblastoma cells were co-cultured with mouse bone marrow cells and primary osteoblasts, ameloblastoma cells induced osteoclast formation. ARL4C elevation in ameloblastoma further promoted its formation capabilities through the increased RANKL expression of mouse bone marrow cells and/or primary osteoblasts. These results suggest that the RAF1-MEK/ERK-ARL4C axis, which may function in cooperation with the BRAFV600E-MEK/ERK pathway, promotes ameloblastoma development. © 2021 The Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Mean flow and turbulence structure over exposed roots on a forested floodplain: Insights from a controlled laboratory experiment.

The time-averaged and instantaneous flow velocity structures of flood waters are not well understood for irregular surfaces such as are created by the presence of roots and fallen branches on forested floodplains. Natural flow structures commonly depart systematically from those described for idealised roughness elements, and an important knowledge gap exists surrounding the effects of natural flow structures on vertical exchanges of fluid and momentum. An improved understanding of the flow structure is required to model flows over forested floodplains more accurately, and to distinguish their dynamics from non-vegetated floodplains or indeed floodplains with other vegetation types, such as reed or grass. Here we present a quantification of the three-dimensional structure of mean flow velocity and turbulence as measured under controlled conditions in an experimental flume using a physical reproduction of a patch of forested floodplain. The results conform in part to existing models of local flow structure over simple roughness elements in aspects such as flow separation downstream of protruding roots, flow reattachment, and the lowering of the velocity maximum further downstream. However, the irregular shape of the surface of the floodplain with exposed roots causes the three-dimensional flow structure to deviate from that anticipated based on previous studies of flows over idealised two-dimensional roughness elements. The results emphasise varied effects of inheritance of flow structures that are generated upstream-the local response of the flow to similar obstacles depends on their spatial organisation and larger-scale context. Key differences from idealised models include the absence of a fully-developed flow at any location in the test section, and various interactions of flow structures such as a reduction of flow separation due to cross-stream circulation and the diversion of the flow over and around the irregular shapes of the roots.

Immunomodulatory role of Keratin 76 in oral and gastric cancer.

Keratin 76 (Krt76) is expressed in the differentiated epithelial layers of skin, oral cavity and squamous stomach. Krt76 downregulation in human oral squamous cell carcinomas (OSCC) correlates with poor prognosis. We show that genetic ablation of Krt76 in mice leads to spleen and lymph node enlargement, an increase in regulatory T cells (Tregs) and high levels of pro-inflammatory cytokines. Krt76-/- Tregs have increased suppressive ability correlated with increased CD39 and CD73 expression, while their effector T cells are less proliferative than controls. Loss of Krt76 increases carcinogen-induced tumours in tongue and squamous stomach. Carcinogenesis is further increased when Treg levels are elevated experimentally. The carcinogenesis response includes upregulation of pro-inflammatory cytokines and enhanced accumulation of Tregs in the tumour microenvironment. Tregs also accumulate in human OSCC exhibiting Krt76 loss. Our study highlights the role of epithelial cells in modulating carcinogenesis via communication with cells of the immune system.

A Public Health Approach in Responding to the Spread of Helium Suicide in Hong Kong.

BACKGROUND: The use of the helium suicide method has been increasing in popularity in Hong Kong since 2012. We have learned a valuable lesson in curbing the spread of charcoal burning (CB) suicide in the past 15 years and hope to prevent the helium suicide method from taking off in the community. AIMS: To document what actions have been taken to contain the spread of the helium suicide method and review the preliminary impact of these actions. METHOD: We adopted a public health approach by engaging stakeholders from multiple sectors, including the police force, the fire services department, coroners, pathologists, mass media, and online media outlets. RESULTS: A monitoring system was established by compiling data extracted from news reports, coroners' reports, and police investigations. Risk and protective factors were identified. Intervention strategies were developed to strengthen protective factors and minimize risk factors. This novel suicide method has not spread as rapidly as the CB suicide method. The preliminary outcomes suggest our actions to be effective. LIMITATIONS: The count of helium suicides in 2015 might be low. The impacts of the interventions are only estimated and require additional empirical verifications. CONCLUSION: The public health approach of engaging multiple partners in the early phase of a potential epidemic can be a good guide to meeting the challenges posed by any new suicide methods that emerge in the future.

Image-based DNA ploidy analysis aids prediction of malignant transformation in oral lichen planus.

OBJECTIVES: To investigate the potential of image-based DNA ploidy analysis to predict malignant transformation in patients with oral lichen planus (OLP). STUDY DESIGN: DNA ploidy analysis was performed on biopsy samples from 14 patients with OLP who underwent malignant transformation. As controls, 42 OLP lesions showing unusual clinical features suggesting a transformation risk and 68 samples of clinically and histologically typical OLP were included. Cases with dysplasia on initial biopsy were excluded. Eighty fibroepithelial polyps acted as methodologic controls. Epithelial nuclei were isolated from formalin-fixed paraffin embedded biopsy samples and monolayers stained with Feulgen for automated image cytometry to establish DNA content. Ploidy status was correlated to outcome using Kaplan-Meier analysis and log-rank Mantel-Cox tests. RESULTS: All controls and typical OLP were diploid and none underwent malignant transformation in mean follow-up of 14 years (10-18 years). One unusual OLP developed carcinoma and all were diploid. The 14 patients with transformation developed 21 carcinomas. In the 11 patients who had a prior biopsy, 4 were aneuploid. CONCLUSIONS: DNA ploidy analysis predicted malignant transformation in more than one third (36.4%) of patients with OLP with a preceding biopsy (n = 11). This premalignant nature could not have been diagnosed clinically or by histologic dysplasia assessment.

High frequency of BRAF V600E mutations in ameloblastoma.

Ameloblastoma is a benign but locally infiltrative odontogenic neoplasm. Although ameloblastomas rarely metastasise, recurrences together with radical surgery often result in facial deformity and significant morbidity. Development of non-invasive therapies has been precluded by a lack of understanding of the molecular background of ameloblastoma pathogenesis. When addressing the role of ERBB receptors as potential new targets for ameloblastoma, we discovered significant EGFR over-expression in clinical samples using real-time RT-PCR, but observed variable sensitivity of novel primary ameloblastoma cells to EGFR-targeted drugs in vitro. In the quest for mutations downstream of EGFR that could explain this apparent discrepancy, Sanger sequencing revealed an oncogenic BRAF V600E mutation in the cell line resistant to EGFR inhibition. Further analysis of the clinical samples by Sanger sequencing and BRAF V600E-specific immunohistochemistry demonstrated a high frequency of BRAF V600E mutations (15 of 24 samples, 63%). These data provide novel insight into the poorly understood molecular pathogenesis of ameloblastoma and offer a rationale to test drugs targeting EGFR or mutant BRAF as novel therapies for ameloblastoma.

Predictive value of dysplasia grading and DNA ploidy in malignant transformation of oral potentially malignant disorders.

Dysplasia grading is widely used to assess risk of transformation in oral potentially malignant disorders despite limited data on predictive value. DNA ploidy analysis has been proposed as an alternative. This study examines the prognostic value for both tests used in a routine diagnostic setting to inform clinical management. A retrospective study of conventional dysplasia grading was conducted on 1,401 patients. DNA ploidy analysis was conducted on a subset of 273 patients and results correlated with clinical information, pathologic diagnosis, and outcome over 5 to 15 years. Malignant transformation occurred in 32 of 273 patients (12%) and, of these, 20 (63%) of preexisting index lesions were aneuploid. Of 241 patients not developing carcinoma, only 39 (16%) of index lesions were aneuploid. Epithelial dysplasia correlated with DNA ploidy status (P < 0.001). The overall positive predictive value for malignant transformation by DNA aneuploidy was 38.5% (sensitivity 65.2% and specificity 75%) and by severe dysplasia grade 39.5% (sensitivity 30% and specificity 98%). DNA diploid and tetraploid status had negative predictive value of 90% to 96%. Combining DNA ploidy analysis with dysplasia grading gives a higher predictive value than either technique alone. Each of three traditional dysplasia grades predicts a significantly different risk of carcinoma development and time to transformation. DNA ploidy analysis had equivalent predictive value and also detected additional risk lesions in the absence of dysplasia.

In vitro studies on odontogenic tumors.

Ameloblastomas are uncommon benign neoplasms of the jaws. They originate from dental epithelial cells, but they are not capable of mineralizing or forming enamel. The study of these tumors is limited to live tissue collected from patients during scheduled surgery. Ameloblastomas grow slowly in vivo and this property is translated to their behavior in vitro. Here, we describe the methods to culture ameloblastomas in organotypic cultures, as well as to isolate stem/progenitor cells from these tumors.

Differentiation-promoting culture of competent and noncompetent keratinocytes identifies biomarkers for head and neck cancer.

Aberrant contact-inhibited proliferation and differentiation induction couple with tumor severity, albeit with an imprecise association with prognosis. Assessment of contact inhibition and differentiation-promoting culture in this study of normal and immortalized oral keratinocytes (NOK and SVpgC2a, respectively) demonstrated elevated cloning ability and saturation density in the immortalized versus normal state, including consistent absence of differentiated morphological features. Transcriptomic analysis implicated 48 gene ontology categories, 8 molecular networks, and 10 key regulator genes in confluency-induced differentiation of NOK, all of which remained nonregulated in SVpgC2a. The SVpgC2a versus NOK transcriptome enriched 52 gene ontology categories altogether, 18 molecular networks, and 39 key regulator genes, several of which were associated with epithelial-mesenchymal transition. Assessment of the previously described gene sets relative to training data sets of head and neck squamous cell carcinoma samples, one including data on tumor differentiation and patient outcome and one present in the Human Gene Expression Map, identified four genes with association to poor survival (COX7A1, MFAP5, MPDU1, and POLD1). This gene set predicted poor outcome in an independent data set of 71 head and neck squamous cell carcinomas. The present study defines, for the first time to our knowledge, the broad gene spectrum that couples to induction, and loss, of oral keratinocyte differentiation. Bioinformatics assessments of the results relative to clinical data generated novel differentiation-related tumor biomarkers relevant to patient outcome.

Squamous cell carcinoma of the tonsillar remnant--clinical presentation and oncological outcome.

BACKGROUND: Squamous cell carcinoma (SCC) of the tonsil is the most common malignant tumour of the oropharynx. Paediatric tonsillectomy is one of the most commonly performed procedures in Otorhinolaryngology. SCC of the tonsil remnant (SCCTR) in a previously tonsillectomised patient is rare. METHODS: Retrospective review of patients with SCCTR presenting to the Otorhinolaryngology, Head and Neck Unit January 2000 to December 2007. RESULTS: Two hundred and fifty patients with tonsil SCC were identified. Ten (4%) of these had previously undergone tonsillectomy in childhood. Nine patients underwent radical treatment including surgery, radiotherapy and in four cases concomitant chemotherapy. Eight patients are alive with no signs of recurrence with follow-up of a minimum of 24 months. One has been lost from follow-up. CONCLUSIONS: Clinicians should be aware that SCC can arise from a tonsillar remnant. SCCTR has similar oncological outcomes as tonsillar tumours.

Oral effects of low-dose methotrexate treatment.

Methotrexate is used increasingly in low-dose regimes for a variety of conditions, particularly rheumatoid arthritis. While certain adverse effects of low-dose methotrexate have been described in detail, oral complications have received little attention. This article includes a summary of the uses and pharmacology of low-dose methotrexate and the mechanisms that lead to general and oral toxicity. The literature relevant to potential oral adverse effects is discussed and 7 illustrative cases are presented. The oral effects noted range from nonhealing ulcers to lymphoma-like lesions. Dental practitioners should be aware of the possible oral effects of low-dose methotrexate that have so far been largely unrecognized.

Analysis of proliferation, apoptosis and keratin expression in cultured normal and immortalized human buccal keratinocytes.

The current study was undertaken to analyse growth and differentiation-related functions of normal keratinocytes (NOK) and an SV40T-immortalized keratinocyte line (SVpgC2a) from buccal mucosa, viewing the latter cell line as a model of a dysplastic epithelium. Morphological and immunohistochemical assessments of organotypic epithelia generated from 10 or 17 d of culture showed three- to five-fold higher apoptotic and proliferative activity in SVpgC2a relative to NOK. Conditions with or without serum (up to 10%) did not significantly influence these parameters in NOK whereas serum supported proliferation of SVpgC2a. Both cell types showed basal expression of collagen IV and laminin 1, indicating basal lamina, as well as vimentin, indicating an activated, proliferative state. Reduced expression of keratin, including the non-keratinizing marker K13, was seen in SVpgC2a. Assessment of proliferative monolayer cultures by microarray showed that NOK transcribed tissue-specific keratins, but also the epidermal keratin K2a, several simple epithelial keratins and low levels of hair keratins. SVpgC2a transcribed keratins seen in epithelial dysplasia, and K2a and hair keratins, albeit at low level. Overall, the results implied aberrant apoptosis, proliferation and keratin expression in the immortalized state of SVpgC2a. Comparison of NOK and SVpgC2a under identical culture conditions may serve to model the progression from a normal to a pre-neoplastic state of buccal epithelium.