Cannabidiol alters mitochondrial bioenergetics via VDAC1 and triggers cell death in hormone-refractory prostate cancer.

In spite of the huge advancements in both diagnosis and interventions, hormone refractory prostate cancer (HRPC) remains a major hurdle in prostate cancer (PCa). Metabolic reprogramming plays a key role in PCa oncogenesis and resistance. However, the dynamics between metabolism and oncogenesis are not fully understood. Here, we demonstrate that two multi-target natural products, cannabidiol (CBD) and cannabigerol (CBG), suppress HRPC development in the TRansgenic Adenocarcinoma of the Mouse Prostate (TRAMP) model by reprogramming metabolic and oncogenic signaling. Mechanistically, CBD increases glycolytic capacity and inhibits oxidative phosphorylation in enzalutamide-resistant HRPC cells. This action of CBD originates from its effect on metabolic plasticity via modulation of VDAC1 and hexokinase II (HKII) coupling on the outer mitochondrial membrane, which leads to strong shifts of mitochondrial functions and oncogenic signaling pathways. The effect of CBG on enzalutamide-resistant HRPC cells was less pronounced than CBD and only partially attributable to its action on mitochondria. However, when optimally combined, these two cannabinoids exhibited strong anti-tumor effects in TRAMP mice, even when these had become refractory to enzalutamide, thus pointing to their therapeutical potential against PCa.

Proposal of a protocol for thermographic assessment of the orbicularis oris muscle.

BACKGROUND: The orbicularis oris muscle is extremely important to performing various oral functions, including mastication, swallowing and speech. Infrared thermography is a non-invasive painless technique that does not require either contrast or ionisation. It has been increasingly used in speech-language-hearing therapy in the last years. OBJECTIVE: The objective of the study was to propose a thermographic analysis method for the orbicularis oris muscle. METHODS: This is an observational, analytical, cross-sectional study. The area of the orbicularis oris muscle was defined based on its anatomy with thermographic images of 11 children and 13 adults. Then, this area was divided into four quadrants, each of which was analysed considering four different shapes: a triangle; a rectangle; a triangle with a rounded side, including the vermilion zone; and a customised shape encompassing only the region of the orbicularis oris muscle, not including the vermilion zone. Data were collected and analysed with the coefficient of variation and interrater agreement. RESULTS: Data variability for the four shapes had similar dispersions per region of the orbicularis oris muscle, in both maximum and mean temperatures and in both children and adults. The rectangle was the shape with the lowest coefficient of variation in more regions of both adults and children. Interrater agreement was excellent for all shapes, in both children and adults. CONCLUSION: Currently, the best way to analyse the orbicularis oris muscle's quadrants is to use the maximum temperature and the rectangle, based on an initial ellipsis encompassing the whole muscle.

Enhancement of hepatic autophagy increases ureagenesis and protects against hyperammonemia.

Ammonia is a potent neurotoxin that is detoxified mainly by the urea cycle in the liver. Hyperammonemia is a common complication of a wide variety of both inherited and acquired liver diseases. If not treated early and thoroughly, it results in encephalopathy and death. Here, we found that hepatic autophagy is critically involved in systemic ammonia homeostasis by providing key urea-cycle intermediates and ATP. Hepatic autophagy is triggered in vivo by hyperammonemia through an α-ketoglutarate-dependent inhibition of the mammalian target of rapamycin complex 1, and deficiency of autophagy impairs ammonia detoxification. In contrast, autophagy enhancement by means of hepatic gene transfer of the master regulator of autophagy transcription factor EB or treatments with the autophagy enhancers rapamycin and Tat-Beclin-1 increased ureagenesis and protected against hyperammonemia in a variety of acute and chronic hyperammonemia animal models, including acute liver failure and ornithine transcarbamylase deficiency, the most frequent urea-cycle disorder. In conclusion, hepatic autophagy is an important mechanism for ammonia detoxification because of its support of urea synthesis, and its enhancement has potential for therapy of both primary and secondary causes of hyperammonemia.

A Rapid Antigen Detection Test to Diagnose SARS-CoV-2 Infection Using Exhaled Breath Condensate by A Modified Inflammacheck® Device.

BACKGROUND: The standard test that identifies the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is based on reverse transcriptase-polymerase chain reaction (RT-PCR) from nasopharyngeal (NP) swab specimens. We compared the accuracy of a rapid antigen detection test using exhaled breath condensate by a modified Inflammacheck® device with the standard RT-PCR to diagnose SARS-CoV-2 infection. METHODS: We performed a manufacturer-independent, cross-sectional, diagnostic accuracy study involving two Italian hospitals. Sensitivity, specificity, positive (PLR) and negative likelihood ratio (NLR), positive (PPV) and negative predictive value (NPV) and diagnostic accuracy with 95% confidence intervals (95% CI) of Inflammacheck® were calculated using the RT-PCR results as the standard. Further RT-PCR tests were conducted on NP specimens from test positive subjects to obtain the Ct (cycle threshold) values as indicative evidence of the viral load. RESULTS: A total of 105 individuals (41 females, 39.0%; 64 males, 61.0%; mean age: 58.4 years) were included in the final analysis, with the RT-PCR being positive in 13 (12.4%) and negative in 92 (87.6%). The agreement between the two methods was 98.1%, with a Cohen's κ score of 0.91 (95% CI: 0.79-1.00). The overall sensitivity and specificity of the Inflammacheck® were 92.3% (95% CI: 64.0%-99.8%) and 98.9% (95% CI: 94.1%-100%), respectively, with a PLR of 84.9 (95% CI: 12.0-600.3) and a NLR of 0.08 (95% CI: 0.01-0.51). Considering a 12.4% disease prevalence in the study cohort, the PPV was 92.3% (95% CI: 62.9%-98.8%) and the NPV was 98.9% (95% CI: 93.3%-99.8%), with an overall accuracy of 98.1% (95% CI: 93.3%-99.8%). The Fagan's nomogram substantially confirmed the clinical applicability of the test in a realistic scenario with a pre-test probability set at 4%. Ct values obtained for the positive test subjects by means of the RT-PCR were normally distributed between 26 and 38 cycles, corresponding to viral loads from light (38 cycles) to high (26 cycles). The single false negative record had a Ct value of 33, which was close to the mean of the cohort (32.5 cycles). CONCLUSIONS: The modified Inflammacheck® device may be a rapid, non-demanding and cost-effective method for SARS-CoV-2 detection. This device may be used for routine practice in different healthcare settings (community, hospital, rehabilitation).

Clinical application of nasal nitric oxide measurement in allergic rhinitis: A systematic review and meta-analysis.

BACKGROUND: Nasal nitric oxide (nNO) is considered a biomarker of nasal inflammation. OBJECTIVE: To perform a systematic review with meta-analysis and meta-regressions on the association between nNO levels and allergic rhinitis (AR). METHODS: PubMed, Web of Science, Scopus, and EMBASE databases were systematically searched. Differences between cases and controls were expressed as standardized mean differences (SMD) with 95% confidence intervals (CI). RESULTS: Overall, 39 articles were included: 30 containing data on nNO measured by nasal aspiration (1881 patients with AR and 1337 controls) and 12 assessing nNO by nasal exhalation (525 patients with AR and 350 controls). Compared with controls, AR presented significantly higher nNO values both during nasal aspiration (SMD, 1.309; 95% CI, 0.841-1.777; P < .001) and nasal exhalation (SMD, 0.708; 95% CI, 0.303-1.114; P = .001). Sensitivity and subgroup analyses confirmed that the results for the evaluated outcomes were not affected by the presence of clinical confounding factors (asthma, nasal polyps, inhaled corticosteroids, smoking history), this being valid for both perennial and seasonal diseases during exposure to allergens. For the aspiration method, meta-regressions indicated that older age and a better pulmonary function were associated with a lower difference in nNO levels between patients with AR and controls, whereas an increasing aspiration flow was associated with a high effect size. CONCLUSION: nNO levels are higher in AR, particularly when using high aspiration flows and in younger patients, who often perceive this condition as a source of disability. Further studies are needed to evaluate the usefulness of this biomarker for monitoring airway disorders and optimizing strategies in different settings (community, hospital, rehabilitation).

Routine blood tests as a potential diagnostic tool for COVID-19.

Objectives The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) to date, the epidemic has gradually spread to 209 countries worldwide with more than 1.5 million infected people and 100,000 deaths. Amplification of viral RNA by rRT-PCR serves as the gold standard for confirmation of infection, yet it needs a long turnaround time (3-4 h to generate results) and shows false-negative rates as large as 15%-20%. In addition, the need of certified laboratories, expensive equipment and trained personnel led many countries to limit the rRT-PCR tests only to individuals with pronounced respiratory syndrome symptoms. Thus, there is a need for alternative, less expensive and more accessible tests. Methods We analyzed the plasma levels of white blood cells (WBCs), platelets, C-reactive protein (CRP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), γ-glutamyl transpeptidase (GGT), alkaline phosphatase and lactate dehydrogenase (LDH) of 207 patients who, after being admitted to the emergency room of the San Raffaele Hospital (Milan, Italy) with COVID-19 symptoms, were rRT-PCR tested. Of them, 105 tested positive, whereas 102 tested negative. Results Statistically significant differences were observed for WBC, CRP, AST, ALT and LDH. Empirical thresholds for AST and LDH allowed the identification of 70% of either COVID-19-positive or -negative patients on the basis of routine blood test results. Conclusions Combining appropriate cutoffs for certain hematological parameters could help in identifying false-positive/negative rRT-PCR tests. Blood test analysis might be used as an alternative to rRT-PCR for identifying COVID-19-positive patients in those countries which suffer from a large shortage of rRT-PCR reagents and/or specialized laboratory.

The influence of tongue mobility on children's performance in computer games that depend on lingual movements.

BACKGROUND: Alteration in tongue mobility requires rehabilitation work through oral motor exercises. These exercises can be integrated with computer games to increase the patient's motivation during treatment. OBJECTIVE: To investigate the influence of tongue mobility on children's motor performance in a computer game reliant on lingual movements. METHODS: A cross-sectional descriptive observational study was carried out with 16 children with altered tongue mobility and 16 children with normal tongue mobility. The subjects were between 8 and 12 years of age. They underwent a clinical evaluation of the tongue and performed an activity using an intra-oral joystick controlled by the tongue to play a simple computer game. The game consisted of targets appearing on the screen that the participants had to reach by moving the joystick control rod. Afterward, the participants answered a feedback questionnaire. Motor performance in the game, measured by the number of reached targets and by the time to reach the targets, was compared between groups and across directions of tongue movement and order of appearance of the targets. RESULTS: The group with altered tongue mobility presented a higher time to reach the target in the downward direction and in the first and last 12 targets and a lower number of targets reached in the left direction, upward direction, and in the first 12 targets than the control group. The direction of the movement influenced tongue performance in both groups. CONCLUSION: Children with altered tongue mobility exhibited a worse performance than those with normal tongue mobility.

The influence of tongue strength on children's performance in computer games reliant on lingual force generation.

Oral motor exercises, for recovering tongue strength, can be integrated with computer games to increase motivation, especially for treatments in children. The aim of this study was to investigate the influence of tongue strength on motor performance in computer games reliant on lingual force generation. An observational study was carried out at a speech-language pathology outpatient university clinic. Twenty participants (10 with normal tongue strength and 10 with reduced tongue strength) aged 8-13 years used an intra-oral joystick controlled by the tongue to play six computer games during which they had to reach targets that appeared on the screen. Motor performance was measured by the number of attempts to score and the time during which the target force was maintained. Tongue motor performance was compared between groups and across directions of tongue movement, resistance force levels, order of target appearance, continuous force application time on the target, age and sex. Children with normal tongue strength had a lower number of attempts to score (P = .014) and maintained the target force for longer periods (P = .002) than those with reduced tongue strength. The performance was better for both groups (a) in the downward direction compared with the performance in other directions, (b) in games with the lowest resistive force level (0.5 N) compared to those with other levels of resistive force and (c) in the second and third rounds compared with the first round. There were no gender-related differences in performance. Older participants performed better than younger participants. Tongue strength, direction of movement, force to reach the target, time of continuous force application, order of target appearance and age influenced tongue motor performance.

NMR Profiling of Exhaled Breath Condensate Defines Different Metabolic Phenotypes of Non-Cystic Fibrosis Bronchiectasis.

Nuclear-magnetic-resonance (NMR) profiling of exhaled breath condensate (EBC) provides insights into the pathophysiology of bronchiectasis by identifying specific biomarkers. We evaluated whether NMR-based metabolomics discriminates the EBC-derived metabolic phenotypes ("metabotypes") of 41 patients with non-cystic fibrosis (nCF) bronchiectasis of various etiology [24 subjects with Primary Ciliary Dyskinesia (PCD); 17 patients with bronchiectasis not associated with PCD (nCF/nPCD)], who were compared to 17 healthy subjects (HS). NMR was used for EBC profiling, and Orthogonal Projections to Latent Structures with partial least-squares discriminant analysis (OPLS-DA) was used as a classifier. The results were validated by using the EBC from 17 PCD patients not included in the primary analysis. Different statistical models were built, which compared nCF/nPCD and HS, PCD and HS, all classes (nCF/nPCD-PCD-HS), and, finally, PCD and nCF/nPCD. In the PCD-nCF/nPCD model, four statistically significant metabolites were able to discriminate between the two groups, with only a minor reduction of the quality parameters. In particular, for nCF/nPCD, acetone/acetoin and methanol increased by 21% and 18%, respectively. In PCD patients, ethanol and lactate increased by 25% and 28%, respectively. They are all related to lung inflammation as methanol is found in the exhaled breath of lung cancer patients, acetone/acetoin produce toxic ROS that damage lung tissue in CF, and lactate is observed in acute inflammation. Interestingly, a high concentration of ethanol hampers cilia beating and can be associated with the genetic defect of PCD. Model validation with 17 PCD samples not included in the primary analysis correctly predicted all samples. Our results indicate that NMR of EBC discriminates nCF/nPCD and PCD bronchiectasis patients from HS, and patients with nCF/nPCD from those with PCD. The metabolites responsible for between-group separation identified specific metabotypes, which characterize bronchiectasis of a different etiology.

An In Vitro Model to Investigate the Role of Helicobacter pylori in Type 2 Diabetes, Obesity, Alzheimer's Disease and Cardiometabolic Disease.

Helicobacter pylori (Hp) is a Gram-negative bacterium colonizing the human stomach. Nuclear Magnetic Resonance (NMR) analysis of intracellular human gastric carcinoma cells (MKN-28) incubated with the Hp cell filtrate (Hpcf) displays high levels of amino acids, including the branched chain amino acids (BCAA) isoleucine, leucine, and valine. Polymerase chain reaction (PCR) Array Technology shows upregulation of mammalian Target Of Rapamycin Complex 1 (mTORC1), inflammation, and mitochondrial dysfunction. The review of literature indicates that these traits are common to type 2 diabetes, obesity, Alzheimer's diseases, and cardiometabolic disease. Here, we demonstrate how Hp may modulate these traits. Hp induces high levels of amino acids, which, in turn, activate mTORC1, which is the complex regulating the metabolism of the host. A high level of BCAA and upregulation of mTORC1 are, thus, directly regulated by Hp. Furthermore, Hp modulates inflammation, which is functional to the persistence of chronic infection and the asymptomatic state of the host. Finally, in order to induce autophagy and sustain bacterial colonization of gastric mucosa, the Hp toxin VacA localizes within mitochondria, causing fragmentation of these organelles, depletion of ATP, and oxidative stress. In conclusion, our in vitro disease model replicates the main traits common to the above four diseases and shows how Hp may potentially manipulate them.

Metabolite variation in three edible Italian Allium cepa L. by NMR-based metabolomics: a comparative study in fresh and stored bulbs.

INTRODUCTION: In fruits and vegetables, comparative analysis of metabolic plant profiles has a high potential for quality control of active components. Onion (Allium cepa L.) is used fresh or stored as food, spice, and in traditional medicine. Its metabolic content, often with nutraceutical value, makes its level an important factor in agronomic production. OBJECTIVE: To describe for the first time the metabolome of "San Pietro" white onion (WP), and compare its chemical profile with the red onion var. Tropea (RT) and the yellow onion var. Montoro (CM). Furthermore, we also aim to obtain a multivariate model based on NMR fingerprints to discriminate the three Italian A. cepa L. cultivars. METHODS: For the chemical fingerprinting we used NMR-based metabolomics. We investigated the aqueous and chloroform extracts of fresh onion at harvesting time, and after 9-month storage. Principal component analysis (PCA), Partial least squares discriminant analysis (PLS-DA) and Orthogonal partial least squares (OPLS-DA) were used to build reliable models. RESULTS: We obtained a clear discrimination of A. cepa L. varieties for the fresh and stored batches. The statistical model highlighted higher levels of fructo-oligosaccharides (FOS) in the fresh WP; RT showed a high content of glucose, citrate and amino acids, while CM had many sulfur components. In the stored samples (CMS, RTS), carbohydrates and sulfur components decreased, while in WPS the free monosaccharides concentration increased. Linoleic acid was overexpressed in the apolar extracts of CMF and WPF cultivars. CONCLUSION: Metabolomics allows a reliable differentiation among onion varieties, and highlights the potential of fingerprinting for food authentication purposes.

Exhaled and nasal nitric oxide measurement in the evaluation of chronic cough.

Chronic cough is one of the most common and troublesome nonspecific respiratory symptom for which patients seek a general practitioner and specialist advice. It is conventionally defined as a cough lasting for more than 8 weeks. Exhaled nitric oxide has proven to be a specific biomarker capable to discriminate between differential diagnoses of chronic cough and simultaneously provide information about the response to specific treatment. In this review, we will discuss the potential use of exhaled and nasal nitric oxide in the diagnosis of chronic chough.

Association between solar ultraviolet doses and vitamin D clinical routine data in European mid-latitude population between 2006 and 2018.

The major source of vitamin D in humans is the ultraviolet radiation-dependent cutaneous synthesis of cholecalciferol; however, low vitamin D status is common in Europe even at mid-latitudes. The UV-radiation that reached the Earth's surface near Milan between May 2006 and December 2018 was retrieved from the TEMIS database and matched with the serum vitamin D levels measured in 30 400 people living in the same area. The results showed a high percentage of insufficient vitamin D levels (measured as 25-hydroxy-vitamin D) throughout the years. During the "vitamin D winter" (November-March) up to 60-90% of the population shows deficient/insufficient (<20-30 ng mL-1) levels of vitamin D and it is explained by the difficulty in obtaining the recommended UV vitamin D doses. In contrast, the warm season provides plenty of UV-radiation, but still 30-50% of the population shows deficient/insufficient vitamin D levels. The circannual vitamin D variations were less evident in the female groups which, in the cold season, show values higher than the corresponding male groups. An age group analysis explained this difference by the strongly recommended vitamin D intake for post-menopausal women. In conclusion, increasing the medical advice for vitamin D intake is strongly recommended to improve the vitamin D status at European mid-latitudes. Our findings suggest that UV availability alone cannot explain the vitamin D status of the population which instead is likely to be influenced by several other factors related to both the people's lifestyle and their personal characteristics. A desirable vitamin D range considering the time of the year and sun exposure, but also including factors not related to UV-radiation, would probably result in a more accurate diagnosis of the patients' vitamin D status. Despite the relatively large time interval, no evident effects due to climate changes were observed in the vitamin D levels during the almost 13 years of analysis.

D2A sequence of the urokinase receptor induces cell growth through αvß3 integrin and EGFR.

The urokinase receptor (uPAR) stimulates cell proliferation by forming a macromolecular complex with αvß3 integrin and the epidermal growth factor receptor (EGFR, ErbB1 or HER1) that we name the uPAR proliferasome. uPAR transactivates EGFR, which in turn mediates uPAR-initiated mitogenic signal to the cell. EGFR activation and EGFR-dependent cell growth are blocked in the absence of uPAR expression or when uPAR activity is inhibited by antibodies against either uPAR or EGFR. The mitogenic sequence of uPAR corresponds to the D2A motif present in domain 2. NMR analysis revealed that D2A synthetic peptide has a particular three-dimensional structure, which is atypical for short peptides. D2A peptide is as effective as EGF in promoting EGFR phosphorylation and cell proliferation that were inhibited by AG1478, a specific inhibitor of the tyrosine kinase activity of EGFR. Both D2A and EGF failed to induce proliferation of NR6-EGFR-K721A cells expressing a kinase-defective mutant of EGFR. Moreover, D2A peptide and EGF phosphorylate ERK demonstrating the involvement of the MAP kinase signalling pathway. Altogether, this study reveals the importance of sequence D2A of uPAR, and the interdependence of uPAR and EGFR.

A new method for tongue rehabilitation with computer games: Pilot study.

BACKGROUND: Lingual exercises are commonly used in clinical practice for swallowing rehabilitation. Associating lingual exercises with computer games increases motivation, which influences tongue motor performance. OBJECTIVE: To investigate the effects of tongue movement direction; resistance force level; repetition number; sustained tongue contraction duration; age and gender on tongue motor performance in healthy adults using computer games. METHODS: An observational pilot study was carried out at a university laboratory. Nine healthy adults, aged 22 to 38 years, used an intra-oral joystick controlled by the tongue to play four computer games. The participants had to reach 12 targets that appeared on the computer screen using the intra-oral joystick. Motor performance was measured by the number of attempts to score and the time during which the target force was maintained. Tongue motor performance was compared among tongue movement direction, resistance force level, game round number, and continuous force application time on the target, age and gender. RESULTS: The number of attempts depended significantly on the direction, continuous force application time on the target and age. The time during which the target force was maintained depended significantly on the direction, continuous force application time on the target and game round number. There were no significant differences in the comparisons by gender or by resistance force level. CONCLUSIONS: It was seen that young adults had their best performance in the downward direction, on the third round, holding the force for a shorter time. The performance deteriorated as age increased.

Coexistence of obesity and asthma determines a distinct respiratory metabolic phenotype.

BACKGROUND: Epidemiologic and clinical evidence supports the existence of an obesity-related asthma phenotype. No distinct pathophysiologic elements or specific biomarkers have been identified thus far, but increased oxidative stress has been reported. OBJECTIVE: We aimed at verifying whether metabolomics of exhaled breath condensate from obese asthmatic (OA) patients, lean asthmatic (LA) patients, and obese nonasthmatic (ONA) subjects could recognize specific and statistically validated biomarkers for a separate "asthma-obesity" respiratory metabolic phenotype, here defined as "metabotype." METHODS: Twenty-five OA patients, 30 ONA subjects, and 30 mild-to-moderate LA age-matched patients participated in a cross-sectional study. Nuclear magnetic resonance (NMR) profiles were analyzed by using partial least-squares discriminant analysis, and the results were validated with an independent patient set. RESULTS: From NMR profiles, we obtained strong regression models that distinguished OA patients from ONA subjects (quality parameters: goodness-of-fit parameter [R2] = 0.81 and goodness-of-prediction parameter [Q2] = 0.79), as well as OA patients from LA patients (R2 = 0.91 and Q2 = 0.89). The all-classes comparison (R2 = 0.86 and Q2 = 0.83) indicated that OA patients possess a respiratory metabolic profile fully divergent from those obtained in the other patient groups. We also identified specific biomarkers for between-class separation, which are independent from clinical bias. They are involved in the methane, pyruvate, and glyoxylate and dicarboxylate metabolic pathways. CONCLUSIONS: NMR-based metabolomics indicates that OA patients are characterized by a respiratory metabolic fingerprint fully different from that of patients independently affected by asthma or obesity. Such a phenotypic difference strongly suggests unique pathophysiologic pathways involved in the pathogenesis of asthma in adult obese subjects. Furthermore, the OA metabotype could define a strategy for patient stratification based on unbiased biomarkers, with important diagnostic and therapeutic implications.

Analytical evaluation of the performances of Diazyme and BRAHMS procalcitonin applied to Roche Cobas in comparison with BRAHMS PCT-sensitive Kryptor.

BACKGROUND: Procalcitonin (PCT) is a recognized marker of sepsis, and its use is expanding to antibiotic stewardship. The aim of this study was the evaluation of two methods: Diazyme PCT on Roche Cobas c702 (PCT-D) and BRAHMS PCT on Roche Cobas e602 analyzers (PCT-BR) in comparison with BRAHMS PCT-sensitive Kryptor (PCT-BK). METHODS: Imprecision was assessed at six critical concentrations following the CLSI EP5-A3; limits of detection (LoDs) were checked according to CLSI EP17-A2; linearity was tested, and method comparison was performed on 239 serum samples. RESULTS: Overall CVs ranged from 12.58% to 5.97% for PCT-D, from 3.94% to 1.70% for PCT-BR and from 6.57% to 1.90% for PCT-BK. LoDs were 0.143 µg/L, 0.014 µg/L, 0.040 µg/L for PCT-D, PCT-BR and PCT-BK, respectively. The functional assay sensitivity was 0.24 µg/L for PCT-D, 0.045 µg/L for PCT-BK and <0.035 µg/L for PCT-BR. PCT-BR was linear up to 68.7 µg/L, PCT-BK up to 43 µg/L and PCT-D up to 27.2 µg/L. Method comparison: PCT-D=0.6543 PCT-BK+0.014, r=0.8463 (but 0.44 if calculated on 0-5 µg/L range); PCT-BR=0.9125 PCT-BK+0.021, r=0.9917. Cohen's κ ranged from 45.2% at 0.25 µg/L to 57.0% at 2.00 µg/L between PCT-D and PCT-BK, whereas it ranged from 89% to 81.3% between PCT-BR and PCT-BK. CONCLUSIONS: The PCT-D performances were significantly different from those of PCT-BR and PCT-BK regarding sensitivity, precision, linearity and agreement at clinical cutoffs. For some patients with serial testing, significantly deviating results were obtained compared to reference. In contrast to Roche PCT assay, it does not seem feasible to use BRAHMS PCT cutoffs for the Diazyme test.

Morphological Simulation of Different Incisal Embrasures: Perception of Laypersons, Orthodontic Patients, General Dentists and Orthodontists.

OBJECTIVE: This study aimed to evaluate how different forms of incisal embrasures influence the esthetic perception of the smile, the preferences of each group of evaluators, and the influence of gingival display. MATERIAL AND METHODS: Two photographs of a broad male and female smiles were digitally manipulated to remove imperfections and asymmetries. A second manipulation was performed on the four maxillary incisors, creating 3 different forms of incisal embrasures: rounded, semi-rounded and squared. The lower region of the upper lip was shifted down to simulate a smile with and without gingival display, resulting in a total of 12 photos. A presentation in digital media was structured with instructions and the photos to be studied by 240 evaluators, who were divided into four groups: laypersons, patients undergoing orthodontic treatment, general dentists and orthodontists. The statistical analysis comprised the Multiple Factor ANOVA (SPANOVA), and the Tukey's post-test was applied at a 5% significance level. RESULTS: The semi-rounded form was generally preferred, with a score of 70.83 for the male smile, and 77.26 for the female smile. Gingival display generated a statistically significant influence on how the embrasures were perceived, with the semi-rounded being preferred when associated with gingival display. Orthodontists were the most stringent in their evaluations. Patients and laypersons exhibited different patterns of esthetic perception. CONCLUSIONS: The form of the incisal embrasures associated with gingival display influenced the evaluators' esthetic perception. There was a statistically significant difference between the groups of patients and orthodontists. CLINICAL SIGNIFICANCE: Incisal embrasures play an important role in smile esthetics and influence the attractiveness perception. In general, the semi-rounded embrasure form is the most preferred. Different groups' perceptions on tooth morphology reinforce the importance of patient participation in treatment planning. (J Esthet Restor Dent 29:68-78, 2017).

Variations between maxillary central and lateral incisal edges and smile attractiveness.

INTRODUCTION: In this study, we aimed to verify whether different levels of maxillary incisal edges influence the perception of smile attractiveness and whether gingival display affects this perception according to groups of orthodontists, dentists, orthodontic patients, and laypersons. METHODS: Photographs of the smiles of 1 man and 1 woman showing the gingival contours of the incisors and the canines were digitally altered, creating steps from 0 to 2.0 mm in 0.5-mm increments, with and without gingival exposure. The 20 pictures were shown in random order to 240 evaluators divided into 4 groups who were asked to provide attractiveness scores on visual analog scales. RESULTS: Both the steps (P <0.001) and the gingival exposure (P <0.05) had statistically significant influences on the evaluations in all groups. There was also a statistically significant difference (P <0.001) between the evaluations of orthodontists and the other groups, with distinct patterns. CONCLUSIONS: The most accepted vertical relationship of incisor borders was the 1.0-mm step. There were significant differences in the evaluation of orthodontists when compared with the other 3 groups, and no significant difference was detected between these groups. The gingival display altered significantly the esthetic perception of the smiles evaluated. There were significant differences between the evaluations of the smiles of the man and the woman.