Evidence that the catalytic mechanism of heme a synthase involves the formation of a carbocation stabilized by a conserved glutamate.

In eukaryotes and many aerobic prokaryotes, the final step of aerobic respiration is catalyzed by an aa3-type cytochrome c oxidase, which requires a modified heme cofactor, heme a. The conversion of heme b, the prototypical cellular heme, to heme o and ultimately to heme a requires two modifications, the latter of which is conversion of a methyl group to an aldehyde, catalyzed by heme a synthase (HAS). The N- and C-terminal halves of HAS share homology, and each half contains a heme-binding site. Previous reports indicate that the C-terminal site is occupied by a heme b cofactor. The N-terminal site may function as the substrate (heme o) binding site, although this has not been confirmed experimentally. Here, we assess the role of conserved residues from the N- and C-terminal heme-binding sites in HAS from prokaryotic (Shewanella oneidensis) and eukaryotic (Saccharomyces cerevisiae) species - SoHAS/CtaA and ScHAS/Cox15, respectively. A glutamate within the N-terminal site is found to be critical for activity in both types of HAS, consistent with the hypothesis that a carbocation forms transiently during catalysis. In contrast, the residue occupying the analogous C-terminal position is dispensable for enzyme activity. In SoHAS, the C-terminal heme ligands are critical for stability, while in ScHAS, substitutions in either heme-binding site have little effect on global structure. In both species, in vivo accumulation of heme o requires the presence of an inactive HAS variant, highlighting a potential regulatory role for HAS in heme o biosynthesis.

Radiotherapy Increases aMMP-8-Levels and Neutrophil/Lymphocyte Ratio Rapidly in Head and Neck Cancer Patients: A Pilot Study.

Radiotherapy for head and neck carcinoma (HNC) has both curative and palliative purposes. This study investigated mouthrinse aMMP-8 levels, molecular forms of MMP-8, blood neutrophil counts and neurophil/lymphocyte ratios before and 3 weeks after HNC radiotherapy started. Thirteen HNC patients undergoing radiotherapy were included. Mouthrinse samples (before and 3 weeks after HNC radiotherapy had started) were assayed quantitatively by aMMP-8 point-of-care-kit (PerioSafe®/ORALyzer®) and by western immunoblot. Total neutrophil counts and neutrophil/lymphocyte ratios were evaluated in the hemogram results. Three weeks after HNC radiotherapy started, significant increases in aMMP-8 levels and neutrophil/lymphocyte ratios were observed. No significant difference was found in total neutrophil counts. Elevations of the activated and fragmented MMP-8 levels after HNC radiotherapy application were observed on western immunoblot analysis. The increase in the aMMP-8 levels and neutrophil/lymphocyte ratios indicate inflammation both locally and systemically suggesting increased risk for periodontitis due to the HNC radiotherapy.

Influence of periodontitis and diabetes on structure and cytokine content of platelet-rich fibrin.

BACKGROUND: Platelet-rich fibrin (PRF) is a second-generation platelet concentrate with multiple applications in wound healing and regeneration in both periodontitis and diabetes. However, the three dimensional (3-D) structure and cytokine content of PRF might be altered in patients suffering from either/both of the chronic inflammatory conditions, ultimately influencing the efficacy of PRF as a biomaterial for regenerative medicine. AIM: The aim of the present study was hence to evaluate the effect of both these chronic inflammatory diseases on the 3-D structure of PRF membrane. An attempt was also made to compare the growth factor content between the plasma and RBC ends of the prepared PRF gel. MATERIALS & METHODS: L-PRF was prepared for twenty participants, healthy (5), periodontitis (5), T2DM (5) and T2DM with periodontitis (5). Porosity and fiber diameter of PRF membranes was visualized under FE-SEM and measured using ImageJ Software. PDGF-BB and TGF-ß1 levels in PRF gel were assessed by ELISA. RESULTS: The average diameter of fibrin fibers under FE-SEM was 0.15 to 0.30 micrometers. Porosity was higher at the plasma end (p = 0.042). Red blood cell (RBC) end of the membrane had thinner fibers arranged in a comparatively more dense and compact structure with smaller porosities. Healthy subjects had the least porous PRF compared to subjects with either/both of the chronic conditions. PDGF-BB levels were similar along all the four groups. TGF-ß1 levels were highest in healthy subjects. DISCUSSION: 3-D structure and growth factor content of PRF are influenced by a person's periodontal and/or diabetic status. The RBC end of the PRF membrane, as compared to the plasma end, has thinner fibers arranged in a comparatively more dense and compact structure with smaller porosities, and hence should be favored during periodontal regenerative procedures. CONCLUSION: Both periodontitis and diabetes have a significant influence on the 3-D structure and growth factor content of PRF produced.

Infective Endocarditis at a Referral Children's Hospital During 19-Year Period: Trends and Outcomes.

BACKGROUND: We noted a recent increase in cases of infective endocarditis (IE) at our institution. The purpose of the study is to examine the incidence, risk factors, microbiology and outcome of IE in our pediatric population. METHODS: Retrospective review of IE cases during 2002-2020 at Children's Hospital of Michigan, Detroit. RESULTS: 68 patients with IE were identified. There was a 2-fold increase in incidence during the 2012-2020 (late period) compared to the 2002-2011 (early period). The most common predisposing conditions were congenital heart disease (CHD) in 39 (57.4%) and central venous catheter (CVC) in 19 (27.9%). CHD was more frequent in the late period (29/43, 67.4%) compared to early period (10/25, 40.0%) (p = 0.042). In CHD patients, palliative or corrective cardiac surgery was performed prior to IE diagnosis in 4/25 (16%) in early period and 23/43 (53.5%) in the late period (p = 0.004). S. aureus was the most common causative organism (35.3%) followed by streptococci (22.1%). Valve replacement or valvuloplasty was performed in 22.1% of patients. Complications occurred in 20 (29.4%). Mortality occurred in 7 (10.3%): 3 had CHD, 3 had CVC and underlying conditions and 1 had fulminant MRSA infection. CONCLUSION(S): The higher incidence of IE during the late period is likely due to an increase in patients with CHD who had undergone prior cardiac surgery. S. aureus was the predominant pathogen in all patients including those with CHD, followed by streptococci. IE in children continues to be associated with high rates of morbidity and mortality.

Photochemistry and Photophysics of Cholesta-5,7,9(11)-trien-3ß-ol in Ethanol.

Cholesta-5,7,9(11)-trien-3ß-ol (9,11-dehydroprovitamin D3, CTL) is used as a fluorescent probe to track the presence and migration of cholesterol in vivo. We recently described the photochemistry and photophysics of CTL in degassed and air-saturated tetrahydrofuran (THF) solution, an aprotic solvent. The zwitterionic nature of the singlet excited state, 1CTL* is revealed in ethanol, a protic solvent. In ethanol, the products observed in THF are accompanied by ether photoadducts and by photoreduction of the triene moiety to four dienes, including provitamin D3. The major diene retains the conjugated s-trans-diene chromophore and the minor is unconjugated, involving 1,4-addition of H at the 7 and 11 positions. In the presence of air, peroxide formation is a major reaction channel as in THF. X-ray crystallography confirmed the identification of two of the new diene products as well as of a peroxide rearrangement product.

Repeatability of dental plaque quantitation by light induced fluorescence technology in current, former, and never smokers.

BACKGROUND: The effects of smoking on the accumulation of dental plaque have not been studied in depth. We compared dental plaque quantitation obtained with a novel light induced fluorescence technology among current, former, and never smokers and verified measurements' repeatability. METHODS: Dental plaque quantitation was objectively assessed by quantitative light induced fluorescence (QLF) technology on three separate study visits in current, former, and never smokers: baseline (day 0), day 7, day 30. Increase in the fluorescence intensity of at least 30% (ΔR30) and 120% (ΔR120) together with the simple oral hygiene (SOH) scoring were considered for analysis. RESULTS: The QLF parameters were highly repeatable in each study group (p < 0.0001, by regression analyses). All QLF parameters showed a significant difference between never smokers and current smokers (p = 0.041 for ΔR30; p = 0.027 for ΔR120; p = 0.04 for SOH). No significant differences were observed between never and former smokers and between current and former smokers except for ΔR120 (p = 0.033). CONCLUSION: Dental plaque measurements by QLF technology were highly reproducible and showed greater plaque formation among current smokers compared to non-smokers. Objective and reproducible quantitation of dental plaque can be a valuable clinical and regulatory science endpoint to investigate the effect of smoking cessation medications, combustion-free tobacco products, and consumer care products on oral health. CLINICAL RELEVANCE: There is a need to objectively evaluate the relationship between smoking and plaque build-up as well as maturation. Current smokers demonstrated greater and more mature plaque buildup when compared to never and former smokers. Differences in plaque build-up and maturation between current, former and non-smokers may be utilized as an effective tool for patient motivation, identifying therapeutic end-points, translational research as well as prognostication. TRIAL REGISTRATION: The study is a pilot study parts of a larger project with registration ID: NCT04649645. As preliminary study, the pilot study referred into this paper started before the larger study registered in ClinicalTrials.gov.

Plasmonic nanoparticle's anti-aggregation application in sensor development for water and wastewater analysis.

Colorimetric sensors have emerged as a powerful tool in the detection of water pollutants. Plasmonic nanoparticles use localized surface plasmon resonance (LSPR)-based colorimetric sensing. LSPR-based sensing can be accomplished through different strategies such as etching, growth, aggregation, and anti-aggregation. Based on these strategies, various sensors have been developed. This review focuses on the newly developed anti-aggregation-based strategy of plasmonic nanoparticles. Sensors based on this strategy have attracted increasing interest because of their exciting properties of high sensitivity, selectivity, and applicability. This review highlights LSPR-based anti-aggregation sensors, their classification, and role of plasmonic nanoparticles in these sensors for the detection of water pollutants. The anti-aggregation based sensing of major water pollutants such as heavy metal ions, anions, and small organic molecules has been summarized herein. This review also provides some personal insights into current challenges associated with anti-aggregation strategy of LSPR-based colorimetric sensors and proposes future research directions.

Association between periodontitis and cognitive impairment in adults.

DESIGN: To summarize the data on association between periodontal diseases and cognitive impairment in adults this systematic review scrutinized various observational studies till September 2021. This review was carried out in accordance with the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA 2020) guidelines. The authors used PECO framework question,: population-Adults (18 years or older), exposure-adults suffering from periodontitis, comparator-adult group without periodontitis, outcome-adults at high risk for cognitive impairment. CASE/CONTROL SELECTION: Search for the literature was conducted on PubMed, Web of Science, and Cumulative Index to Nursing and Allied Health Literature (CINAHL). Search was limited to human studies with no limitation to year of publication prior to September 2021. Search terms used were related to gingiva, oral bacteria like Porphyromonas gingivalis, gum inflammation, periodontitis, dementia, neuroinflammation, cognitive impairment, Alzheimer's disease, Parkinson disease. Following research, all the studies providing association between periodontal diseases and neurodegenerative diseases with quantitative measures were included in the study. Non-human studies, studies on patients below 18 year old, studies related to influence of treatment and in subjects already suffering from neurological disease were excluded. After removing duplicates, eligible studies were identified and data extracted by two reviewers to make ensure inter examiner reliability and to prevent data entry errors. Data from the studies were tabulated as study design, sample characteristics, diagnosis, exposure biomarkers/measures, outcomes and results. DATA ANALYSIS: Methodological quality of studies was assessed by adapted Newcastle-Ottawa scale. Selection of study groups, comparability and exposure/outcome were used as parameters. Case-control and cohort studies were considered as high-quality studies if six or more stars were awarded out of nine maximum stars and four or more stars for cross-sectional studies out of six stars. Comparability among the groups was studied by taking into account primary factors for Alzheimer's disease such as age and sex and secondary factors like hypertension, osteoarthritis, depression, diabetes mellitus, and cerebrovascular disease. For cohort studies, 10 year follow up and dropout of <10% was considered to be successful. RESULTS: A total of 3693 studies were identified by two independent reviewers and finally 11 studies were included in the final analysis. Six cohort studies, three cross-sectional and two case-control studies were included after excluding remaining studies. Bias in studies was assessed by adapted Newcastle-Ottawa Scale. All included studies were of high methodological quality. Association between periodontitis and cognitive impairment was determined by using different criteria like International classification of disease, clinical measurement of periodontitis subjects, inflammatory biomarkers, microbes and antibodies. It was suggested that subjects with chronic periodontitis since 8 years or more, are at a higher risk of having dementia. Clinical measures of periodontal disease like probing depth, clinical attachment loss, alveolar bone loss were found to be positively associated with cognitive impairment. Inflammatory biomarkers and pre-existing elevated levels of serum IgG specific to periodontopathogens was reported to be associated with cognitive impairment. Within the limitations of the study, the authors concluded that though the patients with long-standing periodontitis are at greater risk for developing cognitive impairment by neurodegenerative diseases, the mechanism by which periodontitis can lead to cognitive impairment is still vague. CONCLUSIONS: Evidence suggests a strong association between periodontitis and cognitive impairment. Still further studies should be done to explore the mechanism involved.

Does nonsurgical periodontal therapy improve glycemic control?

DATA SOURCES: The authors searched Medline via Pubmed, EMBASE, Cochrane Database of Systematic Reviews and Scielo. Additionally, grey literature was also searched with no restrictions regarding date of publication and journal up to March 2022. The search was conducted by two pre-calibrated independent reviewers using AMSTAR 2 and PRISMA checklists. Both MeSH terms, relevant free text and their combinations were utilised to conduct the search. STUDY SELECTION: The authors screened the articles on the basis of their titles and abstracts. Duplicates were removed. Full-text publications were evaluated. Any disagreement was resolved by discussion amongst themselves or with a third reviewer. Only the systematic reviews that included RCTs and CCTs were included involving the articles comparing nonsurgical periodontal treatment alone vs no treatment or nonsurgical periodontal treatment with adjunctive therapeutic modality (antibiotics, laser) vs no treatment or nonsurgical periodontal therapy alone. PICO method was used to define the inclusion criteria and changes in glycated haemoglobin post-intervention 3 months was taken as primary outcome. All the articles with the use of adjunctive therapy other than antibiotics (local or systemic) or laser were excluded. The selection was restricted to English only. DATA EXTRACTION AND SYNTHESIS: Data extraction was performed by two reviewers. For each systematic review and each study, mean and standard deviation of glycated hemoglobin level at each follow-up, number of patients both in intervention and control group, type of diabetes, design of study, follow-up period, number of comparisons in meta-analysis, quality assessment of systematic review was assessed by 16 items AMSTAR 2 (Assessment of Multiple Systematic Reviews) and 27 itemed PRISMA (Preferred Reporting Items for Systematic Review and Meta-Analysis) checklist. JADAD scale was used to assess the risk of bias for included RCTs. Q test was used to calculate statistical heterogeneity and percentage of variation by I2 Index. Both Fixed (Mantel-Haenszel [Peto] test) and random (Dersimonian-Laird test) models were used to estimate individual study. Funnel plot and Egger's linear regression methods were used to evaluate publication bias. RESULTS: Following initial electronic and hand search, 1062 articles were screened for title and abstract and 112 articles were considered for full text eligibility. Finally, 16 systematic reviews were considered for qualitative synthesis of results. 16 systematic reviews described 30 unique meta-analyses. Publication bias was assessed in nine out of 16 systematic reviews. Compared to control or non-treatment group, nonsurgical periodontal therapy resulted in statistically significant mean difference of -0.49% HBA1c reduction at 3 months (p = 0.0041), -0.38% (p = 0.0851). The effect of periodontal therapy with antibiotics compared to NSPT alone was not statistically significant (CI -0.32-0.06, 3 months; CI -0.31-0.53, 6 month). The effect of NSPT and laser vs NSPT for HbA1c was not statistically significant (CI -0.73-0.17, 3-4 month). CONCLUSIONS: Based on included systematic reviews and limitations within the study, nonsurgical periodontal therapy is an effective treatment modality in glycaemic control in diabetic patients in terms of HbA1c reduction both at 3 months and 6 months follow-up. The adjunctive therapies like antibiotic administration whether local or systemic and use of lasers with NSPT does not show statistically significant differences as compared to NSPT alone. However, these findings are based on analysis of available literature based on systematic reviews on this subject.

Is periodontal therapy an effective treatment modality for preventing or managing cardiovascular disease in patients with periodontitis?

DATA SOURCES: Cochrane Oral Health Information specialist searched databases: Cochrane Oral Health's Trials Register, Cochrane Central Register of Controlled Trials in the Cochrane diary, MEDLINE Ovid, Embase Ovid, CINAHL EBSCO and Open Grey up to 17 November 2021 without language, publication status or year restriction. Additionally, Chinese Bio Medical Literature Database, China National Knowledge Infrastructure and VIP database were searched up to 4 March 2022. For ongoing trials, the US National Institutes of Health Trials Register, the World Health Organization (WHO) Clinical Trials Registry Platform (up to 17 November 2021), and Sciencepaper Online (up to 4 March 2022) were also searched. A reference list of included studies, hand searching for important journals, and Chinese professional journals in the relevant field was performed until March 2022. STUDY SELECTION: Authors screened the articles on the basis of their titles and abstracts. Duplicates were removed. Full-text publications were evaluated. Any disagreement was resolved by discussion amongst themselves or in consultation with a third reviewer. Only randomised controlled trials assessing the effects of periodontal treatment on participants having chronic periodontitis with cardiovascular disease (CVD) (secondary prevention) or without cardiovascular disease (primary prevention) with minimum follow-up of one year were considered. Patients having known genetic or congenital heart defects, other sources of inflammation, aggressive periodontitis, or were pregnant and/or lactating were excluded. Subgingival scaling and root planning (SRP) with or without combination of systemic antibiotics with or without active remedies were compared with supragingival scaling, mouth rinse, or no periodontal treatment. DATA EXTRACTION AND SYNTHESIS: Data extraction was performed by two independent reviewers in duplicate. A formal, customised pilot-based data extraction form was used to capture data. Overall risk of bias for each study was categorised as low, medium, and high. For trials having missing data or unclear data, clarification from the authors were sought by mail. Testing for heterogeneity was planned by I2 test. For dichotomous data, fixed-effect model (Mantel-Haenszel) was used; and for continuous data, mean difference and 95% confidence intervals were used as measures of treatment effect. For time-to-event data, Peto or inverse variance method was used. Sensitivity and subgroup analysis was planned to test the stability of conclusion. RESULTS: Following initial electronic and hand search, 1690 articles were screened for title and abstract and 82 articles were considered for full-text eligibility. Finally, two studies out of the reported six articles were included in this review for qualitative synthesis of results, and no study was included in the quantitative analysis. Publication bias was determined using funnel plots which were further assessed using dichotomous and continuous outcome. For primary prevention of CVD in participants with periodontitis and metabolic syndrome, one study (165 participants) provided very low certainty evidence. Scaling and root planning plus amoxicillin and metronidazole could reduce incidence of all-cause death (Peto odds ratio [OR] 7.48, 95% confidence interval [CI] 0.15 to 376.98), or all CVD-related death (Peto OR 7.48, 95% CI 0.15 to 376.98). The possibility that scaling and root planning plus amoxicillin and metronidazole could increase cardiovascular events (Peto OR 7.77, 95% CI 1.07 to 56.1) compared with supragingival scaling measured at 12-month follow-up was observed. For secondary prevention of CVD, one pilot study randomised 303 participants to receive scaling and root planning plus oral hygiene instruction or oral hygiene instruction plus a copy of radiographs and recommendation to follow-up with a dentist (community care). As cardiovascular events had been measured for different time periods between 6 and 25 months, and only 37 participants were available with at least one-year follow-up, the data was not sufficiently robust for inclusion in the review. The study did not evaluate all-cause death and all CVD-related death. Conclusions about the effects of periodontal therapy on secondary prevention of CVD were not drawn. CONCLUSIONS: There is very limited evidence assessing the impact of periodontal therapy on the prevention of cardiovascular disease, and it is insufficient to generate any implications for practice. Further trials are needed before reliable conclusions can be drawn.

Photochemistry and photophysics of cholesta-5,7,9(11)-trien-3ß-ol: a fluorescent analogue of cholesterol.

Cholesta-5,7,9(11)-trien-3ß-ol (9,11-dehydroprovitamin D3, CTL) is used as a fluorescent probe to track the presence and migration of cholesterol in vivo. CTL is known to be photochemically active, but little consideration has been given to the formation efficiency and possible toxicity of its photoproducts. In degassed tetrahydrofuran (THF) solution, we isolated the photoproduct of CTL and of its 25-hydroxy derivative (HOCTL), and X-ray crystal structures were obtained for HOCTL and the photorearrangement product. The X-ray crystal structure and its 1H NMR spectrum confirm the product structure as a pentacyclic HOCTL isomer. In the presence of air in THF, endoperoxide formation via [2+4] addition of 1O2* across the B ring of CTL or HOCTL becomes the dominant photoreaction. The UV spectrum and decay kinetics of the triplet state of HOCTL, the precursor of 1O2*, are determined by transient absorption spectroscopy. We confirm the proposed structure of the endoperoxide by X-ray crystallography. Kinetics analysis of quantum yields provides rate constants for photophysical and photochemical events.

Burden and Associated Genotype Patterns of High-Risk Human Papilloma Virus Infection and Cervical Cytology Abnormalities among Women in Central India.

Background: The epidemiology of human papilloma virus (HPV) infection and the pattern of HPV genotype distribution are much-needed parameters to assess the risk of cervical cancer among females. However, due to less availability of data on HPV burden and its genotypes from various geographical regions in India makes cervical cancer screening modalities and vaccination strategies difficult to implement. Objective: The present study was conducted to identify the various genotypes particularly high-risk HPV types in premalignant or malignant cervical lesions. Methods: The study was a hospital-based cross-sectional study wherein 295 symptomatic women were screened by Pap smear and multiplex real-time PCR was performed for HPV genotypes identification in women with abnormal cervical cytology. Results: Out of 295 women, 237 (80.3%), 45 (15.3%), and 13 (4.4%) women had normal Pap smear, squamous cell carcinoma and precancerous cytology, respectively. Among these 58 women having abnormal cervical cytology, HPV was detected in 48 (81.0%) participants. Most common HPV genotypes in our study were HPV 16 (n = 29; 60.4%) followed by mixed infections; i.e., more than one type of HPV was detected (n = 10, 20.8%). HPV 18 was detected only in 6.25%, whereas other high-risk HPV genotypes were found to be 12.5%. Conclusion: HPV positivity was >80% in women having abnormal Pap smear. The prevalence of HPV 18 was found to be much less in Central India, compared to other parts of country. HPV 16 was the most common genotype followed by mixed HPV genotype infections. It is evident from our study that symptomatic women even if having normal Pap smear should be screened for HPV and followed up with periodic Pap smears for detecting any change in cervical cytology, thus preventing cervical cancer in women.

The clinical association between Periodontitis and COVID-19.

OBJECTIVES: The study aimed to clinically assess the association between periodontitis and COVID-19-related outcomes. MATERIAL AND METHODS: Data pertaining to patient demographics, medical history, blood parameters, periodontal clinical examination and aMMP-8 point-of-care diagnostics (both site-level and patient-level) was recorded for eighty-two COVID-19-positive patients. COVID-19-related outcomes such as COVID-19 pneumonia, death/survival, types of hospital admission and need of assisted ventilation were also assessed. RESULTS: Males were predominantly afflicted with COVID-19, with advanced age exhibiting a greater association with the presence of periodontitis. Higher severity of periodontitis led to 7.45 odds of requiring assisted ventilation, 36.52 odds of hospital admission, 14.58 odds of being deceased and 4.42 odds of COVID-19-related pneumonia. The aMMP-8 mouthrinse kit was slightly more sensitive but less specific than aMMP-8 site-specific tests. CONCLUSIONS: Based on the findings of the present study, periodontitis seems to be related to poorer COVID-19-related outcomes. However, within the constraints of this work, a direct causality may not be established. Periodontitis, by means of skewing the systemic condition for a number of comorbidities, may eventually influence COVID-19 outcomes in an indirect manner. CLINICAL RELEVANCE: The study is the first to clinically, and by means of a validated point-of-care diagnostic methodology, assess the association between periodontal health and COVID-19-related outcomes. Assessment of the periodontal status of individuals can aid in the identification of risk groups during the pandemic along with reinforcing the need to maintain oral hygiene and seeking periodontal care.

Do clinical decision support tools have a role to play in dealing with paediatric dental trauma?

Design Randomised controlled trial.Case selection In order to assess baseline knowledge of dental trauma, paediatric dentists and medical students were administered a pre-test, subsequent to which a random assignment was carried out to one of three learning groups: mobile app clinical decision support tools (CDST), print CDST and no CDST, for the purposes of a post-test.Data analysis The correct answers for the pre-test and post-test, time to completion and answers to the demographic survey were subjected to descriptive statistical analyses using IBM SPSS v25.0 for Windows (SPSS, IBM Corp, Armonk, NY). In order to compare the pre- and post-tests for each group, a paired-sample t test was conducted. An independent-sample t test and Pearson χ2 test were utilised to assess for significant differences between paediatric dentists and medical students. Least significant post-hoc and one-way analysis of variance (ANOVA) tests were conducted among the three groups. The level of significance was set at p <0.05.Results Paediatric dentists obtained significantly higher scores on both the pre- and post-tests when compared to medical students (8.57 ± 0.96 vs 4.20 ± 1.58; p <0.001 and 8.37 ± 1.09 vs 4.96 ± 1.99; p <0.001, respectively).No significant difference was noted pertaining to the time taken to complete the tests among both the groups. In both the groups, the highest scores were obtained by those who used the mobile app version of the CDST; these subjects, however, recorded the longest time to complete the post-test (p <0.001).Conclusions When compared to the absence of an aid, both the mobile app and print versions of the CDST improved the diagnosis and management of injuries to the primary dentition. With medical students demonstrating significant improvement in primary dental trauma management with CDST usage, these tools are recommended to enhance the diagnosis and treatment for such patients.

The association between second-hand smoke and the periodontal status of children and adolescents.

Data sources The authors searched Embase, Medline/PubMed, Web of Science, Scopus and LILACS ranging from inception to 23 November 2021. A grey literature search was run utilising OpenGrey along with Google Scholar (most relevant first 200 hits). For the identification of additional articles of relevance, a hand-search was also conducted upon the list of references of all the included studies. No restrictions in terms of date and language were utilised in the search. Controlled vocabulary in the form of free text and MeSH terms was utilised to execute the search strategy.Study selection A screening of the studies potentially satisfying the eligibility criteria was conducted independently by two reviewers on the basis of their abstracts and titles. The records so retrieved were classified under the headings of 'exclude', 'include' and 'uncertain'. Upon the completion of this exercise, an acquisition was made of the full texts of papers judged relevant. Data extraction was performed on all the studies which met the eligibility criteria. Diverging views among the reviewers were resolved with consensus. The judgement of a third reviewer was sought to resolve any disagreements.Data extraction and synthesis Information extraction covered measurement details (cotinine levels of children and/or self-reported questionnaires from children and/or parents); study identification (year of publication, first author and location); distribution according to the status of second-hand smoke and sample characteristics; exposure criteria (according to cotinine levels or severity of parental smoking); analytic approach utilised (multivariate or bivariate analysis); results along with assessment of outcomes (partial- or full-mouth protocol used) and criteria (index type used). Due to only cross-sectional studies being included upon retrieval, the methodological quality assessment was performed utilising an adapted variant of the Newcastle-Ottawa scale. The judgement criteria used included non-response rate, sample size and representativity, confounding control, determination of exposure, outcome assessment and statistical analysis. Evidence tables were built utilising qualitative synthesis in order to ascertain differences in the results and characteristics of the included studies. In case the eligibility criteria were met by three or more papers, the data was subjected to pair-wise comparisons between those unexposed to second-hand smoke with those exposed to it in terms of the specified periodontal outcome measures. Periodontal outcomes were evaluated as continuous measures with the random-effects model. Cochran's Q-test was utilised to assess the heterogeneity and in cases where the heterogeneity was unexplained, a sensitivity analysis was conducted to evaluate whether a single study exerted a significant effect upon the pooled results.Results Six hundred and thirty-seven records were obtained from the search. After screening of the abstracts and titles, eight studies met the eligibility criteria subsequent to the reading of the complete text of 16 reports. One study was conducted in a high socio-demographic index (SDI) country with four in the middle-high, one in the middle and two in the lower-middle SDI countries. Four studies did not provide details pertaining to outcome assessment, while one included a full-mouth and another three, a partial-mouth assessment protocol. Two studies were deemed to be of high quality in terms of methodological quality assessment. Three reports possessed a satisfactory sample size while none of the included studies included data relating to non-response rate. The authors deemed meta-analysis to be possible only in the case of periodontal probing depth (PPD) and gingival index (GI) scores. There was a positive association reported between a high GI score and second-hand smoke exposure. There was, however, no difference present at the subgroup level in terms of the SDI of the country of the study, the age of the cohort as well as the period in which the study was carried out. In contrast, a significant effect was observed for the location of the study data collection in that children from areas with a higher current-smoker prevalence exhibited greater GI scores as a result of increased second-hand smoke exposure. There was no observable difference from the pooled estimate for second-hand smoke exposure compared to the unexposed cohort in terms of PPD. No effects were demonstrable at the subgroup level upon data stratification for the study country SDI and period of data collection. There was greater clinical attachment loss among children who were exposed to second-hand smoke, particularly for those whose fathers were smokers despite adjustments being made for gender, plaque scores and age.Conclusions The systematic review demonstrates a very low level of certainty in evidence to point towards second-hand smoke exposure of children up to 15 years of age leads to higher GI scores as compared to their unexposed counterparts. Even though no differences have been demonstrated in terms of PPD, there is a necessity for further research to draw conclusions on other pertinent periodontal parameters such as bleeding on probing and clinical attachment loss.

Active matrix metalloproteinase-8 (aMMP-8) point-of-care test (POCT) in the COVID-19 pandemic.

INTRODUCTION: Active matrix metalloproteinase (aMMP)-8 utilized in point-of-care testing (POCT) is regarded as a potential biomarker for periodontal and peri-implant diseases. Various host and microbial factors eventually influence the expression, degranulation, levels and activation of aMMP-8. The type of oral fluids (saliva, mouthrinse, gingival crevicular, and peri-implant sulcular fluids [GCF/PISF], respectively) affect the analysis. AREAS COVERED: With this background, we aimed to review here the recent studies on practical, inexpensive, noninvasive and quantitative mouthrinse and GCF/PISF chair-side POCT lateral flow aMMP-8 immunoassays (PerioSafe and ImplantSafe/ORALyzer) and how they help to detect, predict, monitor the course, treatment and prevention of periodontitis and peri-implantitis. The correlations of aMMP-8 POCT to other independent and catalytic activity assays of MMP-8 are also addressed. EXPERT OPINION: The mouthrinse aMMP-8 POCT can also detect prediabetes/diabetes and tissue destructive oral side-effects due to the head and neck cancers' radiotherapy. Chlorhexidine and doxycycline can inhibit collagenolytic human neutrophil and GCF aMMP-8. Furthermore, by a set of case-series we demonstrate the potential of mouthrinse aMMP-8 POCT to real-time/online detect periodontitis as a potential risk disease for coronavirus disease 2019 (COVID-19). The clinical interdisciplinary utilization of aMMP-8 POCT requires additional oral, medical, and interdisciplinary studies.

Phylogenetic analysis of VP7 and VP4 genes of the most predominant human group A rotavirus G12 identified in children with acute gastroenteritis in Himachal Pradesh, India during 2013-2016.

G12 strains are now considered to be the sixth most prevalent human rotaviruses globally. India has introduced rotavirus vaccine Rotavac® into the national immunization program in 2016 and Himachal Pradesh (HP) is the first state to launch it. During epidemiological rotavirus surveillance in HP, predominance of G12 rotaviruses was observed. This study investigated the genetic variability and evolution of HP G12 strains (n = 15) associated with P-genotypes P[6], P[4], and P[8] identified between 2013 and 2016. Phylogenetic analysis of VP7 gene revealed that all characterized G12 strains clustered in lineage-III and diversified into three subclusters indicating that these strains may have originated from three different ancestral G12 strains. The comparative sequence analysis of HP strains with Rotavac® and Rotarix® vaccine strains revealed various amino acid substitutions in epitope regions of VP7 and VP4 proteins especially at the antibody neutralization sites. Only 12/29 VP7 epitope residues and 2/25 VP4 epitope residues were found to be conserved between HP rotavirus strains and vaccine strains. Both long and short electropherotypes were observed in G12P[4] strains, while a single long electropherotype was observed in G12P[6] strains. Children of ≤11 months were significantly infected with G12 rotaviruses. The frequency of vomiting episodes (≥5/day) was significantly higher in children infected with G12 rotavirus strains as compared to non-G12 rotaviruses (p = 0.0405). Our study provides the comprehensive data on clinical characteristics and evolutionary pattern of the G12 rotavirus, the most prevalent strain in HP and emphasizes the need to monitor these strains for inclusion in future vaccine.

Is saltwater mouth rinse as effective as chlorhexidine following periodontal surgery?

Design Randomised prospective double-blind study.Case selection Thirty-seven chronic periodontitis patients were randomly prescribed either saltwater (n = 17) or chlorhexidine (n = 20) mouth rinse following open flap debridement. Gingival Index (GI), post-operative pain, mouth rinse satisfaction, matrix metalloproteinase activity and tasting were assessed at baseline, one week and 12 weeks.Data analysis Inter-group and intra-group comparisons were done for all the parameters recorded at different time intervals using Friedman, Wilcoxon signed-rank and Mann-Whitney U tests. P value less than 0.05 was considered statistically significant.Results There was a statistically significant decrease in GI from baseline to week 1/week 12 following surgery in both the groups. However, there was no significant difference between the groups at any time point.Conclusions Saltwater rinses are as efficient as 0.12% chlorhexidine in reducing inflammation following minimal invasive periodontal surgery. Being an inexpensive and easily accessible option, it might be considered the mouth rinse of choice during the early stages of wound healing.

Epidemiology and clinical features of rotavirus, adenovirus, and astrovirus infections and coinfections in children with acute gastroenteritis prior to rotavirus vaccine introduction in Meerut, North India.

There are limited reports on the etiology of multiple enteric viruses causing acute gastroenteritis (AGE) in North India. In the present study we have determined the prevalence of three enteric viruses, namely rotavirus, astrovirus (AstV) and adenovirus (AdV) in a total of 312 diarrheic children (<5 years) hospitalized at Lala Lajpat Rai Memorial Medical College, Meerut, Uttar Pradesh from August 2014 to July 2016; and results were compared with data from Delhi. The fecal samples were individually screened for group A rotavirus (RVA), AdV, and AstV using enzyme immunoassay kits. At least one viral agent was detected in 29.2% of 312 fecal specimens. RNA of rotavirus antigen-positive samples was extracted by TRIzol method. Rotavirus G/P genotyping was performed using seminested multiplex reverse transcriptase-polymerase chain reaction. RVA was the most predominant virus (18.3%) followed by AstV (12.5%), and AdV (9.9%). Coinfections were detected in 10.6% cases and the most common coinfection in diarrheic children was RVA combined with AstV (36.4%). Overall, the enteric viruses were found most prevalent in the 6 to 11 months age group (P = .01). Increased duration of vomiting (≥3 days) was significantly (P = .04) associated with AdV infection (61.3%) as compared with AstV (30.76%) and rotavirus (26.31%). G1P[8] was detected throughout as the most prevalent rotavirus strain (10.5%). Unusual RV strains like G2P[6] and G2P[8] were also detected. Of note G3, G4, and G12 rotavirus were detected for the first time in Meerut. This is the first report that demonstrated the important contribution of multiple enteric viruses causing AGE in young children in this part of Uttar Pradesh (Meerut).

SMN deficiency in severe models of spinal muscular atrophy causes widespread intron retention and DNA damage.

Spinal muscular atrophy (SMA), an autosomal recessive neuromuscular disease, is the leading monogenic cause of infant mortality. Homozygous loss of the gene survival of motor neuron 1 (SMN1) causes the selective degeneration of lower motor neurons and subsequent atrophy of proximal skeletal muscles. The SMN1 protein product, survival of motor neuron (SMN), is ubiquitously expressed and is a key factor in the assembly of the core splicing machinery. The molecular mechanisms by which disruption of the broad functions of SMN leads to neurodegeneration remain unclear. We used an antisense oligonucleotide (ASO)-based inducible mouse model of SMA to investigate the SMN-specific transcriptome changes associated with neurodegeneration. We found evidence of widespread intron retention, particularly of minor U12 introns, in the spinal cord of mice 30 d after SMA induction, which was then rescued by a therapeutic ASO. Intron retention was concomitant with a strong induction of the p53 pathway and DNA damage response, manifesting as γ-H2A.X positivity in neurons of the spinal cord and brain. Widespread intron retention and markers of the DNA damage response were also observed with SMN depletion in human SH-SY5Y neuroblastoma cells and human induced pluripotent stem cell-derived motor neurons. We also found that retained introns, high in GC content, served as substrates for the formation of transcriptional R-loops. We propose that defects in intron removal in SMA promote DNA damage in part through the formation of RNA:DNA hybrid structures, leading to motor neuron death.