Detection of a C4 Criegee Intermediate: Fourier-Transform Microwave Spectroscopy of Methacrolein Oxide.

Pure rotational transitions of methacrolein oxide (MACRO) were observed by Fourier-transform microwave spectroscopy. Among the four low-lying conformers existing within an energy window of 3 kcal/mol, only the lowest-energy conformer, the anti-trans conformer, was detected in a discharged jet of a 1,3-diiode-2-methylprop-1-ene and O2 mixture diluted in Ar. Nineteen pure rotational transitions, in the frequency range from 10 to 25 GHz, most of them showing A/E splitting due to the methyl-top internal rotation, were observed and analyzed by the XIAM program, yielding the internal rotation barrier of 559 cm-1, which very well agrees with a theoretically calculated value, 558 cm-1, at the CCSD(T)/cc-pVTZ level of theory.

Gas-phase spectroscopic identification of the chlorovinyl radical.

Fourier transform microwave spectra for two isomers of the chlorine substituted vinyl radical have been observed in the 4-52 GHz frequency region. The observed radicals (2A') have been generated using electric discharges of diluted dichloro derivatives of ethylene as molecular precursors. Fine and hyperfine components observed for each rotational transition are fully assigned in the present study to two isotopologues (35Cl and 37Cl), and precise molecular constants are determined for both radicals.

In vitro model systems for exploring oral biofilms: From single-species populations to complex multi-species communities.

Numerous in vitro biofilm model systems are available to study oral biofilms. Over the past several decades, increased understanding of oral biology and advances in technology have facilitated more accurate simulation of intraoral conditions and have allowed for the increased generalizability of in vitro oral biofilm studies. The integration of contemporary systems with confocal microscopy and 16S rRNA community profiling has enhanced the capabilities of in vitro biofilm model systems to quantify biofilm architecture and analyse microbial community composition. In this review, we describe several model systems relevant to modern in vitro oral biofilm studies: the constant depth film fermenter, Sorbarod perfusion system, drip-flow reactor, modified Robbins device, flowcells and microfluidic systems. We highlight how combining these systems with confocal microscopy and community composition analysis tools aids exploration of oral biofilm development under different conditions and in response to antimicrobial/anti-biofilm agents. The review closes with a discussion of future directions for the field of in vitro oral biofilm imaging and analysis.

Convolution Neural Networks and Targeted Fluorescent Nanoparticles to Detect and ICDAS Score Caries.

Previous work has shown targeted fluorescent starch nanoparticles (TFSNs) can label the subsurface of carious lesions and assist dental professionals in the diagnostic process. In this study, we aimed to evaluate the potential of using artificial intelligence (AI) to detect and score carious lesions using ICDAS in combination with fluorescent imaging following application of TFSNs on teeth with a range of lesion severities, using ICDAS-labeled images as the reference standard. A total of 130 extracted human teeth with ICDAS scores from 0 to 6 were selected by a calibrated cariologist. Then, the same surface was imaged with a stereomicroscope under white light illumination, without visible fluorescence, and blue light illumination with an orange filter following application of the TFSNs. Both sets of images were labeled by another blinded ICDAS-calibrated cariologist to demarcate lesion position and severity. Convolutional neural networks, state-of-the-art models in imaging AI, were trained to determine the presence, location, ICDAS score (severity), and lesion surface porosity (as an indicator of activity) of carious lesions, and tested by 30 k-fold validation for white light, blue light, and the combined image sets. The best models showed high performance for the detection of carious lesions (sensitivity 80.26%, PPV 76.36%), potential for determining the severity via ICDAS scoring (accuracy 72%, SD 5.67%), and the detection of surface porosity as an indicator of the activity of the lesions (accuracy 90%, SD 7.00%). More broadly, the combination of targeted biopolymer nanoparticles with imaging AI is a promising combination of novel technologies that could be applied to many other applications.

Tooth Age Impact on Dental Erosion Susceptibility and Treatment Efficacy.

This laboratory study investigated the impact of tooth age on dental erosion susceptibility and preventive treatment efficacy. Extracted human premolars were selected and had their age estimated (∼10-100 years old) using established dental forensic methods. Enamel and root dentin slabs were prepared, embedded in acrylic blocks, flattened, and polished. The specimens were randomly assigned to one of three treatments (n = 93): Sn+F (800 ppm Sn as SnCl2 and 250 ppm F as NaF, pH 4.5), NaF (250 ppm F, pH 4.5), or deionized water (DIW). Each specimen was subjected for 10 days to a daily cycling protocol consisting of six 5-min erosive challenges (0.3% citric acid, pH 2.6), six 60-min remineralization periods (artificial saliva), and three 2-min treatments with the test solutions. Surface loss (SL) was measured after 3, 5, and 10 days, using optical profilometry. Effects of tooth age, antierosive treatment, and time on SL were evaluated using linear mixed effects regression analysis. SL increased with age for all substrate-treatment-time combinations (p < 0.0001). Sn+F and NaF solutions significantly reduced SL compared to DIW, regardless of substrate, time, or age (p < 0.0001), with best results shown for Sn+F. Efficacy of Sn+F increased with tooth age on enamel, but tooth age did not affect the efficacy of NaF on enamel. For dentin, increased efficacy was observed with age after 5 (for Sn+F) and 10 days (for Sn+F and NaF). In conclusion, increase of tooth age rendered enamel and root dentin more susceptible to dental erosion. NaF preventive efficacy improved with tooth age for dentin, in advanced erosion simulation. Sn+F reduced enamel SL due to erosion regardless of tooth age.

Use of tocilizumab in kidney transplant recipients with COVID-19.

Acute respiratory distress syndrome associated with coronavirus infection is related to a cytokine storm with large interleukin-6 (IL-6) release. The IL-6-receptor blocker tocilizumab may control the aberrant host immune response in patients with coronavirus disease 2019 (COVID-19) . In this pandemic, kidney transplant (KT) recipients are a high-risk population for severe infection and showed poor outcomes. We present a multicenter cohort study of 80 KT patients with severe COVID-19 treated with tocilizumab during hospital admission. High mortality rate was identified (32.5%), related with older age (hazard ratio [HR] 3.12 for those older than 60 years, P = .039). IL-6 and other inflammatory markers, including lactic acid dehydrogenase, ferritin, and D-dimer increased early after tocilizumab administration and their values were higher in nonsurvivors. Instead, C-reactive protein (CRP) levels decreased after tocilizumab, and this decrease positively correlated with survival (mean 12.3 mg/L in survivors vs. 33 mg/L in nonsurvivors). Each mg/L of CRP soon after tocilizumab increased the risk of death by 1% (HR 1.01 [confidence interval 1.004-1.024], P = .003). Although patients who died presented with worse respiratory situation at admission, this was not significantly different at tocilizumab administration and did not have an impact on outcome in the multivariate analysis. Tocilizumab may be effective in controlling cytokine storm in COVID-19 but randomized trials are needed.

Rotational spectra of van der Waals complexes: pyrrole-Ne and pyrrole-Ne2.

The van der Waals 1 : 1 and 1 : 2 adducts between the aromatic molecule pyrrole (Pyr) and the rare gas atom neon (Ne) have been investigated using a combination of chirped pulse Fourier transform microwave spectroscopy and quantum-chemical calculations. Rotational spectra of two and three isotopologues of Pyr-Ne and Pyr-Ne2, respectively, arising from the combinations of the 20Ne and 22Ne isotopes, were identified and a partial rs structure determined. Unusual spectral intensities have been observed with a significant enrichment of heavier isotopic species in the jet molecular expansion. The observed rotational constants of Pyr-Ne are consistent with a nearly symmetric prolate top with the Ne atom located above the plane of pyrrole. The trimer presents C2v symmetry with the Ne atoms located one on each side of the ring plane. The experimental 14N nuclear quadrupole coupling constants have been determined for all the isotopologues of Pyr-Ne and Pyr-Ne2 complexes. Similar values to those of isolated pyrrole have been found, which suggests that the electrical gradient field of pyrrole does not change much upon complexation. The observed spectroscopic parameters have been compared with those of other aromatic-rare gas complexes.

Probing Criegee intermediate reactions with methanol by FTMW spectroscopy.

Criegee intermediates (CIs) are carbonyl oxides generated from ozonolysis of unsaturated hydrocarbons in the atmosphere. The relatively long lifetime of CIs makes possible the bimolecular reactions with other atmospheric agents. These reactions can potentially be fast enough to contribute significantly to the tropospheric budgets of those species or to alter the rate of generation of secondary organic aerosols (SOAs). In particular, the new adducts formed in these reactions contribute SOA formation because they have larger molecular weights and lower vapor pressures than the reactants and are more condensable. α-alkoxyalkyl hydroperoxides are the nascent products derived from the insertion reactions of CIs with alcohols. In this work we report the direct detection of methoxymethyl hydroperoxide (HOOCH2OCH3, MMHP) and methoxyethyl hydroperoxide (HOOC(CH3)HOCH3, MEHP) as the reaction products between the CIs, CH2OO and CH3CHOO and methanol. High resolution Fourier transform microwave spectroscopy has been used to identify one and two conformers of MMHP and MEHP, respectively. The pre-reactive complex CH2OO-CH3OH was not observed in this experiment, which indicates that CH2OO shows a different reactivity toward methanol than that toward water. Our results for CH3CHOO + CH3OH show that MEHP is produced in similar ratios when syn- and anti-CH3CHOO react with methanol vapor.

Observation of hydroperoxyethyl formate from the reaction between the methyl Criegee intermediate and formic acid.

Rapid reactions of carboxylic acids with Criegee intermediates provide efficient gas phase removal processes and are proposed to trigger the formation of new molecular compounds, which are implicated in the growth of atmospheric aerosols. The new adducts formed in these reactions are known as hydroperoxide esters and are predicted to be condensable, with lower vapor pressures than the reactants. We report here the direct detection of hydroperoxyethyl formate (HOOCH(CH3)OCHO) formed in the gas phase reaction between the methyl substituted Criegee intermediate, CH3CHOO, and formic acid. Two different isomers of this hydroperoxide ester have been observed using the combination of pulsed Fourier-transform microwave spectroscopy with isotopic substitution experiments and high-level quantum chemical calculations. The identification of hydroperoxyethyl formate as the main nascent product of the CH3CHOO and HCOOH reaction differs from previous results that reported the formation of vinyl hydroperoxide as the reaction product of this reaction.

Intrinsic folding of the cysteine residue: competition between folded and extended forms mediated by the -SH group.

A dual microwave and optical spectroscopic study of a capped cysteine amino acid isolated in a supersonic expansion, combined with quantum chemistry modelling, enabled us to characterize the conformational preferences of Cys embedded in a protein chain. IR/UV double resonance spectroscopy provided evidence for the coexistence of two conformers, assigned to folded and extended backbones (with classical C7 and C5 backbone H-bonding respectively), each of them additionally stabilized by specific main-chain/side-chain H-bonding, where the sulfur atom essentially plays the role of H-bond acceptor. The folded structure was confirmed by microwave spectroscopy, which demonstrated the validity of the DFT-D methods currently used in the field. These structural and spectroscopic results, complemented by a theoretical Natural Bond Orbital analysis, enabled us to document the capacity of the weakly polar -CH2-SH side chain of Cys to adapt itself to the intrinsic local preferences of the peptide backbone, i.e., a γ-turn or a ß-sheet extended secondary structure. The corresponding local H-bonding bridges the side chain acceptor S atom to the backbone NH donor site of the same or the next residue along the chain, through a 5- or a 6-membered ring respectively.

Introducing BAIT (Biofilm Architecture Inference Tool): a software program to evaluate the architecture of oral multi-species biofilms.

Biofilm model systems are used to study biofilm growth and predict the effects of anti-biofilm interventions within the human oral cavity. Many in vitro biofilm model systems use a confocal laser scanning microscope (CLSM) in conjunction with image analysis tools to study biofilms. The aim of this study was to evaluate an in-house developed image analysis software program that we call BAIT (Biofilm Architecture Inference Tool) to quantify the architecture of oral multi-species biofilms following anti-biofilm interventions using a microfluidic biofilm system. Differences in architecture were compared between untreated biofilms and those treated with water (negative control), sodium gluconate ('placebo') or stannous fluoride (SnF2). The microfluidic system was inoculated with pooled human saliva and biofilms were developed over 22 h in filter-sterilized 25 % pooled human saliva. During this period, biofilms were treated with water, sodium gluconate, or SnF2 (1000, 3439 or 10 000 p.p.m. Sn2+) 8 and 18 h post-inoculation. After 22 h of growth, biofilms were stained with LIVE/DEAD stain, and imaged by CLSM. BAIT was used to calculate biofilm biovolume, total number of objects, surface area, fluffiness, connectivity, convex hull porosity and viability. Image analysis showed oral biofilm architecture was significantly altered by 3439 and 10 000 p.p.m. Sn2+ treatment regimens, resulting in decreased biovolume, surface area, number of objects and connectivity, while fluffiness increased (P<0.01). In conclusion, BAIT was shown to be able to measure the changes in biofilm architecture and detects possible antimicrobial and anti-biofilm effects of candidate agents.

Direct Quantification of Gadolinium Retention in Young Patients by ICP-MS Analysis of Extracted Teeth.

In this 10-patient prospective pilot study, we show the feasibility of pragmatic direct ex vivo measurement of gadolinium retention from group II gadolinium-based contrasts agents (GBCAs) in young patients after routine tooth extraction. This noninvasive method may support future research attempting to understand the link between GBCA exposure and clinical outcomes.

The Criegee intermediate-formic acid reaction explored by rotational spectroscopy.

The atmospheric reaction of the simplest Criegee intermediate, CH2OO, with formic acid has been investigated in the gas phase by pulsed Fourier-transform microwave spectroscopy. The dominant nascent product from this reaction was identified as hydroperoxymethyl formate (HOOCH2OCHO), for which two different conformations, formed through independent insertion mechanisms, were observed in the discharged plasma of a CH2I2/O2/formic acid gas mixture. The conformational identifications are supported by the observation of 13C species in natural abundance together with the chemically mono substituted deuterium isotopologues. These isotopic observations further suggest that hydroperoxymethyl formate slightly decomposes, producing formic anhydride (OHCOCHO) in a dehydration reaction.

Fourier transform microwave spectroscopy of Criegee intermediates: The conformational behaviour of butyraldehyde oxide.

Four conformers of the n-propyl-substituted Criegee intermediate, CH3CH2CH2CHOO, also named n-butyraldehyde oxide, have been observed by Fourier transform microwave spectroscopy. The transient species was produced using a pulsed electric discharge of a gas mixture of 1,1-diiodobutane/O2 diluted in Ar or Ne. The observation of only syn species is in contrast to the results of other previous studies of alkyl-substituted Criegee intermediates, in particular, those for the structural isomers, isobutyraldehyde oxide ((CH3)2CHCHOO), and methyl-ethyl-ketone oxide (C2H5C(CH3)OO), for which syn and anti species have been observed coexisting in the gas phase.

Alkaline and Alkaline-earth Cyanoacetylides: A Combined Theoretical and Rotational Spectroscopic Investigation.

The metallic cyanoacetylides LiC3N, NaC3N, MgC3N and CaC3N have been investigated by combined spectroscopy measurements and theoretical calculations. The theoretical calculations predict for the four species that the linear isomer with formula MCCCN (M= Li, Na, Mg and Ca) is the most stable one. We used the laser ablation molecular beam Fourier transform microwave spectroscopy to synthesize these species by the reaction of metal vapors, produced by laser ablation, and the 3-bromo-2-propynenitrile (BrCCCN). The pure rotational spectra were observed by Fourier transform microwave spectroscopy in the 2-18 GHz frequency region only for LiCCCN and NaCCCN, while no spectral signatures for MgCCCN and CaCCCN could be detected. Finally, we have searched for LiCCCN and NaCCCN species towards the carbon-rich evolved star IRC + 10216 but only upper limits to their abundances have been obtained.

European Organization for Caries Research Workshop: Methodology for Determination of Potentially Available Fluoride in Toothpastes.

Toothpastes are the most universally accepted form of fluoride delivery for caries prevention. To provide anti-caries benefits, they must be able to release fluoride during the time of tooth brushing or post brushing into the oral cavity. However, there is no standard accepted procedure to measure how much fluoride in a toothpaste may be (bio) available for release. The European Organization for Caries Research proposed and supported a workshop with experts in fluoride analysis in toothpastes and representatives from industry. The objective of the workshop was to discuss issues surrounding fluoride analysis in toothpaste and reach consensus on terminology and best practices, wherever the available evidence allowed it. Participants received a background paper and heard presentations followed by structured discussion to define the problem. The group also reviewed evidence on the validity, reliability and feasibility of each technique (namely chromatography and fluoride electroanalysis) and discussed their strengths and limitations. Participants were able to reach a consensus on terminology and were also able to identify and summarize the advantages and disadvantages of each technique. However, they agreed that most currently available methods were developed for regulatory agencies several decades ago, utilizing the best available data from clinical trials then, but require to be updated. They also agreed that although significant advances to our understanding of the mechanism of action of fluoride in toothpaste have been achieved over the past 4 decades, this clearly is an extraordinarily complex subject and more work remains to be done.

Bleaching of simulated stained-remineralized caries lesions in vitro.

OBJECTIVE: Non-invasive esthetic treatment options for stained arrested caries lesions have not been explored. This study aimed to develop laboratory models to create stained-remineralized caries-like lesions (s-RCLs) and to test the efficacy of bleaching on their esthetic treatment. MATERIALS AND METHODS: One hundred twelve enamel/dentin specimens were prepared from human molars, embedded, and had their color measured spectrophotometrically at baseline and after demineralization. They were randomly divided into four groups (n = 14) based on the staining/remineralization protocols for a total of 5 days: G1, no staining/no remineralization; G2, no staining/remineralization in artificial saliva (AS); G3, non-metallic staining/remineralization with sodium fluoride/AS; and G4, metallic staining/remineralization with silver diamine fluoride/AS. The lesion mineral loss (ΔZ) and depth (L) were measured using transverse microradiography along with color change (ΔE). Specimens were bleached and color was re-evaluated. Data were analyzed using ANOVA models followed by Fisher's PLSD tests (α = 0.05). RESULTS: s-RCLs in G4 were significantly (p < 0.001) darker than G3, G2, and G1 regardless of substrate type and condition. s-RCLs in G2, G3, and G4 showed significantly lower ΔZ and L than G1 (all p < 0.001), confirming occurrence of remineralization. G4 exhibited significantly lower ΔZ and L compared to G2 (p < 0.001). Bleaching was more effective in non-metallic than in metallic stained lesions regardless of substrate type (p < 0.001). CONCLUSION: The proposed models created distinct s-RCLs. Non-metallic s-RCLs were lighter and more responsive to bleaching compared to metallic s-RCLs. CLINICAL RELEVANCE: The developed experimental models allow the further investigation of the efficacy and safety of different clinical strategies for the esthetic management of s-RCLs.

Unveiling the Neutral Forms of Glutamine.

Neutral glutamine has been evaporated by laser ablation of its solid sample to seed a rare gas carrier prior to a supersonic expansion and proved by Fourier transform microwave techniques. We report on three distinct neutral conformers that show a singular non-interacting and flexible amide sidechain in contrast with the other proteinogenic aliphatic amino acids. It could explain the essential biological role of glutamine as a nitrogen source, and its unique ability to form a variety of hydrogen bonds with peptide backbones. Common computational methods fail to predict the delicate balance of intramolecular interactions controlling the geometry of the most stable conformer. The spectroscopic data here reported can be used to benchmark novel computational methods in quantum chemistry.

The reaction between the methyl Criegee intermediate and hydrogen chloride: an FTMW spectroscopic study.

Experiments using pulsed Fourier-transform microwave spectroscopy have been performed to probe the reaction between the methyl substituted Criegee intermediate, CH3CHOO, and hydrogen chloride. Two conformers of the product derived from the insertion of HCl within CH3CHOO, chloroethyl hydroperoxide, formed in the discharged plasma of a CH3CHI2/O2/HCl gas mixture, were identified through their experimentally determined rotational and quadrupole coupling constants. The relative abundance of the detected species indicates that the reaction of the anti-CH3CHOO conformer is faster than that of the syn-CH3CHOO conformer. The results presented in this work constitute the first experimental information about the reaction between CH3CHOO and HCl.

The reactivity of the Criegee intermediate CH3CHOO with water probed by FTMW spectroscopy.

The reaction of Criegee intermediates with water is one of the dominant removal mechanisms for these species in the atmosphere. The reactivity of alkyl substituted Criegee intermediates has been shown to be affected by the nature and location of the substituents. CH3CHOO, acetaldehyde oxide, can be considered as a prototypical Criegee intermediate with a single alkyl substituent to examine the conformer specific reactivity for Criegee intermediates. Pulsed Fourier-transform microwave spectroscopy has been used to probe the products resulting from the reaction between CH3CHOO and water. The hydrogen-bonded complex between CH3CHOO and water together with the reaction product, hydroxyethyl hydroperoxide, were observed in the discharged plasma of a CH3CHI2/O2/water gas mixture. The experimentally determined rotational parameters support the identification of the complex between water and the syn-CH3CHOO conformer and two conformers of hydroxyethyl hydroperoxide, produced from the anti-CH3CHOO conformer and water.