Investigating the perceptions and experiences of Canadian dentists on dental regulatory bodies' communications and guidelines during the COVID-19 pandemic.

OBJECTIVE: Dental regulatory bodies aim to ensure the health and safety of dentists, dental staff patients and the public. An important responsibility during a pandemic is to communicate risk and guidelines for patient care. Limited data exist on the perceptions and experiences of dentists navigating new guidelines for mitigating risk in dental care during the pandemic. The objective of this study was to use a qualitative approach to explore how dentists in Canada experienced and perceived their regulatory bodies' communication about COVID-19 risks and guidelines during the pandemic. METHODS: Participants were Canadian dentists (N = 644) recruited through the email roster of nine provincial dental associations or regulatory bodies. This qualitative analysis was nested within a prospective longitudinal cohort study in which data were collected using online questionnaires at regular intervals from August 2020 to November 2021. To address the objective reported in this paper, a conventional qualitative content analysis method was applied to responses to three open-ended questions included in the final questionnaire. RESULTS: Participants encountered challenges and frustrations amid the COVID-19 pandemic, grappling with diverse regulations and communications from dental bodies. While some bodies offered helpful guidance, many participants felt the need for improved communication on guidelines. Dentists urged for expedited, clearer and more frequent updates, expressing difficulty in navigating overwhelming information. Negative views emerged on the vague and unclear communication of COVID-19 guidelines, contributing to confusion and frustration among participants. CONCLUSION: As COVID-19 persists and in planning for future pandemics, these experiential findings will help guide regulatory bodies in providing clear, timely and practical guidelines to protect the health and safety of dentists, dental staff, patients and the public.

Validity and reliability of the COVID-19 Anxiety Syndrome Scale in Canadian dentists.

BACKGROUND: The COVID-19 pandemic has resulted in a high level of mental health problems for the population worldwide including healthcare workers. Several studies have assessed these using measurements for anxiety for general populations. The COVID-19 Anxiety Syndrome Scale (C-19ASS) is a self-report measure developed to assess maladaptive forms of coping with COVID-19 (avoidance, threat monitoring and worry) among a general adult population in the United States. We used it in a prospective cohort study of COVID-19 incidence rates in practising Canadian dentists. We therefore need to ensure that it is valid for dentists in French and English languages. This study aimed to evaluate the validity of the C-19ASS in that population. METHODS: Cross-sectional data from the January 2021 monthly follow-up in our prospective cohort study were used. Exploratory factor analysis (EFA) and confirmatory factor analysis (CFA) were performed. RESULTS: The results of EFA revealed a 2-factor structure solution that explained 47% of the total variance. The CFA showed a good model fit on the data in both English and French languages. The Cronbach's alpha indicated acceptable levels of reliability. Furthermore, the C-19ASS showed excellent divergent validity from the Generalized Anxiety Disorder-7 (GAD-7) scale. CONCLUSIONS: The C-19ASS is valid and reliable instrument to measure COVID-19-related anxiety in English and French among Canadian dentists. PRACTICAL IMPLICATIONS: This validated measure will contribute to understanding of the mental health impact of the pandemic on dentists in Canada and enable the dental regulatory authorities and organizations to intervene to help dentists.

Occupation as a measure of life course socioeconomic position and the risk of oral cancers in India.

OBJECTIVES: Evidence suggests that different indicators of socioeconomic position (SEP) contribute to oral cancer risk. Occupational status, as a measure of SEP, may be able to capture aspects of social hierarchy in societies in which employment is highly correlated with other social structures such as caste systems. Often in such societies, the life course of an individual is also influenced by this hierarchy. However, the influence of life course occupational status on the risk of oral cancer is not well understood. This study aims to identify the life course model that is best supported by the data using life course SEP-as represented by occupation-on oral cancer risk in a population in the South of India. METHODS: Data from the HeNCe Life study, Indian site were used. Incident oral cancer cases (N = 350) were recruited from two major referral hospitals in Kozhikode, Kerala, South India, from 2008 to 2012. Controls (N = 371), frequency-matched by age (5 years) and sex were recruited from the outpatient clinics at the same hospitals as the cases. Life grid-based structured interviews collected information on an array of exposures throughout the life course of the participant. Occupation was coded with the 1988 International Standard Classification of Occupations, transformed to the simplified European Socioeconomic Classification, and further dichotomized into advantageous and disadvantageous SEP at three different life periods (childhood, early adulthood and late adulthood). The analysis was conducted using the Bayesian relevant life course exposure model with a Dirichlet noninformative prior and a weakly informative Cauchy prior to the overall lifetime effect and confounders. RESULTS: Participants in disadvantaged SEP throughout their life had 3.6 times higher risk of oral cancer than those in advantaged SEP (OR = 3.6; 95% CrI = 1.6-7.2), after adjusting for potential confounders. While the crude and sex- and age-adjusted models showed a clear childhood sensitive period for this risk, the model further adjusted for behavioural factors could not distinguish the specific life course period best explained by data. CONCLUSION: Occupation status alone could provide a similar overarching risk estimate for oral cancer to those obtained from more complex measures of SEP.

Do cutaneous human papillomavirus genotypes affect head and neck cancer? Evidence and bias-correction from a case-control study.

BACKGROUND: Three genera of human papillomavirus (HPV) infect the oral cavity and oropharynx- alpha (α), beta (ß) and gamma (γ). While α-HPV infection is an established risk factor for head and neck cancers (HNC), the role of other genera remains unclear. We aimed to estimate the effect of α-, ß-, γ-HPV on HNC using a hospital-based case-control study. METHODS: We recruited incident HNC cases (396) and controls (439), frequency-matched by age and sex from four main referral hospitals in Montreal, Canada. We collected information on sociodemographic and behavior characteristics using in-person interviews, and tested rinse, brush and tumor specimens for HPV genotypes. We estimated adjusted odds ratios (aOR) and 95% confidence intervals (CI) for the effect of HPV on HNC using logistic regression, adjusting for confounding. We conducted probabilistic bias analysis to account for potential exposure misclassification, selection bias, and residual confounding. RESULTS: α-HPV genus had a strong effect on HNC, particularly HPV16 (aOR=22.6; 95% CI: 10.8, 47.2). ß-HPV was more common among controls (aOR=0.80; 95% 0.57, 1.11). After adjustment for HPV16, we found weaker evidence for γ-HPV (aOR= 1.29; 95% CI: 0.80, 2.08). Combined bias analyses for HPV16 increased the strength of the point estimate, but added imprecision (aOR=54.2, 95% CI: 10.7, 385.9). CONCLUSIONS: α-HPV, especially HPV16, appears to increase the risk for HNC, while there is little evidence for an effect of ß- or γ-HPV. ß-HPV may have a preventive effect, while γ-HPV may increase the risk of HNC, although to a lesser extent than that of α-HPV. Results for cutaneous HPV were imprecise and less conclusive. Due to possible epidemiologic biases, the true relation between HPV and HNC could be underestimated in the literature. Further improvement in current methods and more studies of the biologic mechanisms of the three genera in HNC development are warranted.

Differential impact of socioeconomic position across life on oral cancer risk in Kerala, India: An investigation of life-course models under a time-varying framework.

OBJECTIVES: The incidence of oral cancer has been rapidly increasing in India, calling for evidence contributing to a deeper understanding of its determinants. Although disadvantageous life-course socioeconomic position (SEP) is independently associated with the risk of these cancers, the explanatory mechanisms remain unclear. Possible pathways may be better understood by testing which life-course model most influences oral cancer risk. We estimated the association between life-course SEP and oral cancer risk under three life-course models: critical period, accumulation and social mobility. METHODS: We recruited incident oral cancer cases (N = 350) and controls (N = 371) frequency-matched by age and sex from two main referral hospitals in Kozhikode, Kerala, India, between 2008 and 2012. We collected information on childhood (0-16 years), early adulthood (17-30 years) and late adulthood (above 30 years) SEP and behavioural factors along the life span using interviews and a life-grid technique. Odds ratios (OR) and 95% confidence intervals (CI) were estimated for the association between life-course SEP and oral cancer risk using inverse probability weighted marginal structural models. RESULTS: Relative to an advantageous SEP in childhood and early adulthood, a disadvantageous SEP was associated with oral cancer risk [(OR = 2.76, 95% CI: 1.99, 3.81) and (OR = 1.84, 95% CI: 1.21, 2.79), respectively]. In addition, participants who were in a disadvantageous (vs advantageous) SEP during all three periods of life had an increased oral cancer risk (OR = 4.86, 95% CI: 2.61, 9.06). The childhood to early adulthood social mobility model and overall life-course trajectories indicated strong influence of exposure to disadvantageous SEP in childhood on the risk for oral cancer. CONCLUSIONS: Using novel approaches to existing methods, our study provides empirical evidence that disadvantageous childhood SEP is critical for oral cancer risk in this population from Kerala, India.

No role for human papillomavirus infection in oral cancers in a region in southern India.

Oral cancer is a major public health issue in India with ∼ 77,000 new cases and 52,000 deaths yearly. Paan chewing, tobacco and alcohol use are strong risk factors for this cancer in India. Human papillomaviruses (HPVs) are also related to a subset of head and neck cancers (HNCs). We examined the association between oral HPV and oral cancer in a sample of Indian subjects participating in a hospital-based case-control study. We recruited incident oral cancer cases (N = 350) and controls frequency-matched by age and sex (N = 371) from two main referral hospitals in Kerala, South India. Sociodemographic and behavioral data were collected by interviews. Epithelial cells were sampled using Oral CDx® brushes from the oral cancer site and the normal mucosa. Detection and genotyping of 36 HPV genotypes were done using a polymerase chain reaction protocol. Data collection procedures were performed by qualified dentists via a detailed protocol with strict quality control, including independent HPV testing in India and Canada. HPV DNA was detected in none of the cases or controls. Associations between oral cancer and risk factors usually associated with HPV infection, such as oral sex and number of lifetime sexual partners, were examined by logistic regression and were not associated with oral cancer. Lack of a role for HPV infection in this study may reflect cultural or religious characteristics specific to this region in India that are not conducive to oral HPV transmission. A nationwide representative prevalence study is needed to investigate HPV prevalence variability among Indian regions.

Maternal and paternal contribution to intergenerational psychosocial transmission of paan chewing.

OBJECTIVES: Paan chewing is a recognized risk factor for oral cancer in the Asian population. However, there is currently little evidence about the intergenerational psychosocial transmission of paan chewing in South Indian families. We investigated the association between parental and participant's paan chewing in a South Indian population. METHODS: A subset of data was drawn from a hospital-based case-control study on oral cancer, the HeNCe Life study, conducted at Government Dental and Medical Colleges of Kozhikode, South India. Analyses were based on 371 noncancer control participants having diseases unrelated to known risk factors for oral cancer. Demographics, behavioral habits (e.g., paan chewing, smoking), and indicators of socioeconomic position (SEP) of both participants and their parents were collected with the use of a questionnaire-based interview and a life grid technique. Unconditional logistic regression assessed odds ratios (OR) and 95% confidence intervals (95% CI) for the associations between parental and participant's paan chewing, adjusted for confounders. RESULTS: Over half of the participants were males (55.2%), and the mean age of participants was 59 (SD = 12) years. After adjusting for age, religion, parents' SEP, parents' education, smoking and alcohol consumption, and perceived parenting behavior, we observed that maternal paan chewing and paternal paan chewing were significantly associated with the participant's paan chewing ([OR = 2.40, 95% CI = 1.11-5.21] and [OR = 3.05, 95% CI = 1.48-6.27], respectively). CONCLUSIONS: Intergenerational psychosocial transmission of the habit of paan chewing could occur through shared sociocultural or environmental factors.

Novel functional association of rat testicular membrane-associated cytosolic glutathione S transferases and cyclooxygenase in vitro.

AIM: To analyze the role of cytosolic glutathione S-transferases cGSTs and membrane-associated cytosolic GSTs macGSTs in prostaglandin biosynthesis and to evaluate the possible interaction between glutathione S-transferases GSTs and cyclooxygenase (COX) in vitro. METHODS: SDS-PAGE analysis was undertaken for characterization of GSTs, thin layer chromatography (TLC) to monitor the effect of GSTs on prostaglandin biosynthesis from arachidonic acid (AA) and spectrophotometric assays were done for measuring activity levels of COX and GSTs. RESULTS: SDS-PAGE analysis indicates that macGSTs have molecular weights in the range of 25-28 kDa. In a coupled assay involving GSTs, arachidonic acid and cyclooxygenase-1, rat testicular macGSTs produced prostaglandin E2 and F2alfa, while the cGSTs caused the generation of prostaglandin D2, E2 and F2alfa. In vitro interaction studies on GSTs and COX at the protein level have shown dose-dependent inhibition of COX activity by macGSTs and vice versa. This effect, however, is not seen with cGSTs. The inhibitory effect of COX on macGST activity was relieved with increasing concentrations of reduced glutathione (GSH) but not with 1-chloro 2,4-dinitrobenzene (CDNB). The inhibition of COX by macGSTs, on the other hand, was potentiated by glutathione. CONCLUSION: We isolated and purified macGSTs and cGSTs from rat testis and analyzed their involvement in prostaglandin biosynthesis. These studies reveal a reversible functional interaction between macGSTs and COX in vitro, with possible interactions between them at the GSH binding site of macGSTs.

Evidence for the association of synaptotagmin with glutathione S-transferases: implications for a novel function in human breast cancer.

OBJECTIVE: To analyze the pattern of changes in GSTs in cancerous and adjacent non-cancerous tissues obtained from breast cancer patients undergoing surgery. DESIGN AND METHODS: Cytosolic GST purification, assay of GST, protein expression levels, and GST-synaptotagmin association were analyzed using standard biochemical techniques like GSH-affinity purification, spectrophotometry, SDS-PAGE, Western blots, and matrix-assisted laser desorption and ionization-time of flight (MALDI-TOF). RESULTS: GST activity in cancerous tissues (0.26 U/mg protein) was significantly higher (P < 0.05) as compared to those from adjacent non-cancerous tissues (0.14 U/mg protein) of breast cancer patients. Further analysis of GST subunits on SDS-PAGE and Western blots using class-specific GST antibodies revealed significant elevation in GST-pi levels in cancer tissues with no appreciable changes in GST-alpha and GST-mu. Along with the elevation of GST-pi levels, high molecular weight proteins (approximately 70 kDa) cross reacting with GST antibodies were detected only in surgically resected tumor biopsies but not in the non-cancerous tissues adjacent to the tumor. Based on MALDI-TOF analysis, the high molecular weight band was identified as synaptotagmin V bound to GST-M1 with 47% sequence coverage after processing on an MS-FIT search engine. CONCLUSIONS: Our results suggest a novel putative functional role for the GST-synaptotagmin complex in human breast cancers. As this association of GST M1-synaptotagmin was not seen in adjacent non-cancerous tissues, this can be used as a marker for breast cancers.

Expression of cyclooxygenase-2 in rat testis.

Cyclooxygenase, the rate-limiting enzyme in the production of prostaglandins, exists in two isoforms, the constitutive cyclooxygenase 1 (Cox-1) and the inducible cyclooxygenase 2 (Cox-2). Cox-1 is involved in homeostatic functions while Cox-2 is implicated in various pathological processes such as inflammation and cancer. The present study describes the constitutive expression of Cox-2 in immature and mature rat testis, primarily localized in the spermatogonial cells. An interesting observation is the presence of Cox-2 on the chromatin and also in cytosol, apart from nuclear and endoplasmic reticular membranes. The significance of this observation is not yet clear, though its presence in the nucleus raises the possibility of Cox-2 having a more direct role in gene regulation than was thought earlier. In addition, the Cox-2 mRNA in testis is the smaller (2.8 kb) of the two isoform transcripts reported for Cox-2. Further hormone treatment regimes (testosterone/follicle stimulating hormone) increased the levels of Cox-2 protein within 6 h after treatment, suggesting that the sustained levels of Cox-2 protein in testis can be further influenced by gonadotrophins and androgens.

A modified protocol for quantitative fit testing using the PortaCount.

A modified quantitative fit testing method has been developed for testing half masks using the TSI PortaCount respirator fit tester. This approach focuses on shortening the time for each exercise during fit testing; however, the shortened protocol is applied only to the very good-fitting masks. For marginal-fitting masks, the testing is carried out according to the full Occupational Safety and Health Administration (OSHA) respiratory protection standard (29CFR1910.134).(1) The shortened protocol (currently not approved by OSHA) still uses all the exercises required by the OSHA standard but for a shorter time (30 seconds [s] for each exercise instead of the usual 60 s). How good the fit has to be to qualify for a shortened exercise is determined by the statistical analysis of a large data set containing pass and fail fit-test data. The statistical analysis involves calculating the sensitivity and specificity of the pass and failed fit tests on half masks. From this analysis, a multiplication factor (K) to the OSHA pass/fail criterion was developed. For a respirator to undergo the shortened protocol, the fit factor obtained during any exercise must be K times the OSHA pass/fail criterion of 100 for half masks. Hence, this approach is more conservative than fit testing protocols that involve shortened exercises regardless of the fit. Nevertheless, this approach still saves time without compromising the accuracy of the fit test expressed in terms of sensitivity and specificity. For the existing data, 85 percent of the fit tests would have been performed according to the faster test protocol while only 15 percent of the tests would have been tested according to the full-length OSHA test protocol.

New experimental studies of the basic performance characteristics of aerosol samplers.

The study of the basic physical performance characteristics of aerosol samplers, like those used in the occupational hygiene setting, will provide insights to enable the improved development of new instruments and cost-effective testing procedures. These will be required as the new particle size-selective sampling criteria become the basis of new occupational exposure standards. A new body of work is being conducted, in which the factors influencing sampler performance are being investigated using idealized samplers of spherical shape in small wind tunnels. By the experimental methods described, a large amount of performance data can be acquired in a very short time. The results for wide ranges of particle size, wind speed, sampling flow rate, and sampler orientation conditions show that there are strong trends as functions of these variables. Those trends are very complicated. But it is encouraging that they are broadly consistent with recent semi-empirical models, suggesting that extensions of that type of modelling approach--supported by the large amount of new experimental data now being generated by such experiments--might provide new models of aerosol sampler performance accessible to researchers and occupational hygienists.

Antisense GADD45 expression results in decreased DNA repair and sensitizes cells to u.v.-irradiation or cisplatin.

Loss of p53 function in cancer cells commonly results in a condition of genomic instability. This is believed to emanate from a loss of the G1 checkpoint response to DNA damage. While the role of p53 in the induction of a G1 arrest is well-accepted, additional p53 functions are being discovered. Cell cycle checkpoints presumably function to allow additional time for DNA repair after damage is incurred, however, genetic studies in yeast suggest that components of the checkpoint pathway may also be involved in DNA lesion processing (Lydall and Weinert, 1995). Recent evidence suggests that this may also be the case for p53, as suggested by numerous reports linking p53 function to DNA repair. Thus, loss of p53 function might contribute to genomic instability independent of G1-arrest. In the present study, we explored the effect of p53 disruption and consequences of antisense GADD45 expression on the DNA repair capacity of human colon carcinoma RKO cells. DNA repair was assayed using host-cell reactivation of u.v.-damaged reporter plasmids and unscheduled DNA synthesis experiments in transiently-transfected cells. We show that a number of transfected genes that suppress p53 function reduce the ability of cells to repair u.v.-induced DNA damage. Moreover, cells in which expression of the p53-regulated gene GADD45 was blocked by antisense vectors, also showed altered levels of DNA repair. Blocking Gadd45 expression by constitutive antisense expression sensitized cells to killing by u.v.-radiation or by cis-platinum (II) diamine-dichloride (CDDP, or cisplatin), a cancer chemotherapy drug which produces DNA cross-links. These findings suggest the involvement of downstream effectors of the p53 pathway in the coordination of cell cycle arrest and DNA repair.

Quantitation of mitochondrial DNA in human lymphoblasts by a competitive polymerase chain reaction method: application to the study of inhibitors of mitochondrial DNA content.

With increasing awareness of the mitochondrial toxicity associated with certain 2',3'-dideoxynucleosides used in anti-human immunodeficiency virus therapy, procedures for quantitative analyses of drug effects on mitochondrial DNA (mtDNA) have assumed enhanced importance. For this reason we have developed a method to measure the copy numbers of mtDNA in cultured MOLT-4 cells. First a hybrid competitive DNA template was synthesized by conventional polymerase chain reaction (PCR), using two custom-synthesized 40-mer composite primers incorporating mitochondrial displacement loop sequences linked by a non-mitochondrial cDNA template (a 76-base pair sequence from the tat/rev region of human immunodeficiency virus cDNA). For the competitive assay, increasing known copy numbers of the hybrid competitive template were added as an internal control to samples containing total cellular DNA. With this approach, two competitive PCR products were generated, 1) a mitochondrial displacement loop-derived fragment (182 base pairs) and 2) a competitive DNA template-derived fragment (156 base pairs). Absolute quantitation was achieved by radiometric comparison of the relative amounts of the two products. To test the versatility of this method, varying amounts of competitive template (6.6 x 10(4) to 6.6 x 10(9) copies) were used with a fixed quantity of total cellular DNA taken from cells cultured for 9 days in the presence or absence of selected pyrimidine and purine dideoxynucleosides. The results showed that the copy number of cellular mtDNA is 823 +/- 71 copies/cell in MOLT-4 cells. Little selective depletion of mtDNA, compared with total cellular DNA, was seen with the purine dideoxynucleosides examined; however, when the cells were exposed to the pyrimidine dideoxynucleoside 2',3'-dideoxycytidine (50 nM) for 9 days, mtDNA content was specifically depleted, although total cellular DNA decreased by only 10%. Thus, in addition to the presently used methods of assessing mitochondrial impairment, i.e., Southern blot analysis and electron microscopy, the competitive PCR method provides a third and convenient assay, with particular applicability to determination of mtDNA in very small numbers of cells.

Comparison of biochemical parameters of benzamide riboside, a new inhibitor of IMP dehydrogenase, with tiazofurin and selenazofurin.

The biochemical and cytotoxic activities of the IMP dehydrogenase (IMPDH) inhibitors benzamide riboside, tiazofurin, and selenazofurin were compared. These three C-nucleosides exert their cytotoxicity by forming an analogue of NAD, wherein nicotinamide is replaced by the C-nucleoside base. The antiproliferative activities of these three agents were compared in a panel of 60 human cancer cell lines. To examine the relationship of benzamide riboside and selenazofurin to tiazofurin, COMPARE computer analysis was performed, and correlation coefficients of 0.761 and 0.815 were obtained for benzamide riboside and selenazofurin, respectively. The biochemical activities of these agents were examined in human myelogenous leukemia K562 cells. Incubation of K562 cells for 4 hr with 10 microM each of benzamide riboside, selenazofurin and tiazofurin resulted in a 49, 71, and 26% decrease in IMPDH activity with a concurrent increase in intracellular IMP pools. As a consequence of IMPDH inhibition, GTP and dGTP concentrations were curtailed. These studies demonstrated that selenazofurin was the most potent of the three agents. To compare the cellular synthesis of NAD analogues of these agents, K562 cells were incubated with 10 microM each of benzamide riboside, tiazofurin and selenazofurin after prelabeling the cells with [2,8-3H]adenosine. The results demonstrated that benzamide riboside produced 2- and 3-fold more of NAD analogue (BAD) than tiazofurin and selenazofurin did. To elucidate the effects of the three compounds on other NAD-utilizing enzymes, the inhibitory activities of purified benzamide adenine dinucleotide (BAD), thiazole-4-carboxamide adenine dinucleotide (TAD) and selenazole-4-carboxamide adenine dinucleotide (SAD) were studied in commercially available purified preparations of lactate dehydrogenase, glutamate dehydrogenase and malate dehydrogenase. TAD and SAD did not inhibit these three dehydrogenases. Although BAD did not influence lactate and glutamate dehydrogenases, it selectively inhibited 50% of malate dehydrogenase activity at a 3.2 microM concentration. These studies demonstrate similarities and differences in the biochemical actions of the three C-nucleosides, even though they share similar mechanisms of action.