Effect of topical fluoride gel and fluoride varnish in the prevention of white spot lesions in patients undergoing fixed orthodontic treatment: An in vivo study

Objectives: Our primary objective was to establish the efficacy of fluoride gel and fluoride varnish in the prevention of white spot lesions (WSLs) development during fixed orthodontic treatment. Materials and Methods: The study sample consisted of 60 adult patients in a prospective split-mouth study design. Interventions, that is, topical fluoride gel and topical fluoride varnish were assigned at the time of bonding to either the right or left halves of the dentition. In all subjects, repeated evaluation of demineralization was done on the facial surfaces of sample teeth in each quadrant. Evaluation using laser fluorescence and by direct visual observation under magnification was carried out at bonding (T0), 3 months (T1), and 6 months (T2). Results: The distribution of mean DIAGNOdent score at T1 (3.14 ± 1.00 vs. 2.81 ± 0.852) and T2 (4.17 ± 1.41 vs. 3.51 ± 1.13) was observed which is significantly higher in the Gel group compared to the Varnish group. In the Gel group, the distribution of mean DIAGNOdent score at T1 (3.14 ± 1.00) and T2 (4.17 ± 1.41) is significantly higher compared to mean DIAGNOdent score at baseline T0 (2.07 ± 0.66). In the Varnish group, the distribution of mean DIAGNOdent score at T1 (2.81 ± 0.852) and T2 (3.51 ± 1.13) is significantly higher compared to the mean DIAGNOdent score at T0 (2.07 ± 0.66). Visual scores also correlated with DIAGNOdent scores. Conclusion: Fluoride varnish is more efficacious than fluoride gel in reducing enamel demineralization. Initial application of fluoride varnish around the orthodontic bracket at bonding appointment can offer significant protection against WSLs.

LanM Peptides – Unravelling the Binding Properties of the EF-Hand Loop Sequences Stripped from the Structural Corset

Since the discovery of the biological relevance of lanthanides (Lns) for methylotrophic bacteria in the last decade, the field has seen a steady rise in discoveries of bacteria using Lns. The major role of lanthanides here is in the active sites of enzymes: methanol dehydrogenases. Additionally, lanthanide binding proteins have also been identified. One such protein is lanmodulin (LanM) and, with a remarkable selectivity for Lns over Ca(II) and affinities in the picomolar range, it makes an attractive target to address challenges in lanthanide separation. Why LanM has such a high selectivity is currently not entirely understood, both the specific amino acid sequences of the EF-hand loops, together with cooperativity effects have been suggested. Consequently, we decided to remove the effect of cooperativity by focusing on the amino acid level. Thus, we synthesized all four 12-amino acid EF-Hand loop peptides of LanM using solid phase peptide synthesis and investigated their affinity for Lns (Eu(III), Tb(III)), the actinide Cm(III) and Ca(II). Using isothermal titration calorimetry and time resolved laser fluorescence spectroscopy combined with parallel factor analysis, we show that in the absence of cooperativity the short EF-Hand loop peptides have all similar affinities for lanthanides and that these are all in the micromolar range. Furthermore, calcium was shown not to bind to the peptides which was verified with circular dichroism spectroscopy. This technique also revealed that the peptides undergo a change to a more ordered state when lanthanides are added. These experimental observations were further supported by molecular dynamics simulations. Lastly, we put Eu(III) and Cm(III) in direct competition using TRLFS. Remarkably, a slightly higher affinity for the actinide, as was also observed for LanM, was found. Our results demonstrate that the picomolar affinities in LanM are largely an effect of pre-structuring in the full protein and therefore reduction of flexibility in combination with cooperative effects, and that all EF-Hand loops possess similar affinities when detached from the protein backbone, albeit still retaining the high selectivity for lanthanides and actinides over calcium.

Effect of Temperature and Cell Viability on Uranium Biomineralization by the Uranium Mine Isolate Penicillium simplicissimum

The remediation of heavy-metal-contaminated sites represents a serious environmental problem worldwide. Currently, cost- and time-intensive chemical treatments are usually performed. Bioremediation by heavy-metal-tolerant microorganisms is considered a more eco-friendly and comparatively cheap alternative. The fungus Penicillium simplicissimum KS1, isolated from the flooding water of a former uranium (U) mine in Germany, shows promising U bioremediation potential mainly through biomineralization. The adaption of P. simplicissimum KS1 to heavy-metal-contaminated sites is indicated by an increased U removal capacity of up to 550 mg U per g dry biomass, compared to the non-heavy-metal-exposed P. simplicissimum reference strain DSM 62867 (200 mg U per g dry biomass). In addition, the effect of temperature and cell viability of P. simplicissimum KS1 on U biomineralization was investigated. While viable cells at 30°C removed U mainly extracellularly via metabolism-dependent biomineralization, a decrease in temperature to 4°C or use of dead-autoclaved cells at 30°C revealed increased occurrence of passive biosorption and bioaccumulation, as confirmed by scanning transmission electron microscopy. The precipitated U species were assigned to uranyl phosphates with a structure similar to that of autunite, via cryo-time-resolved laser fluorescence spectroscopy. The major involvement of phosphates in U precipitation by P. simplicissimum KS1 was additionally supported by the observation of increased phosphatase activity for viable cells at 30°C. Furthermore, viable cells actively secreted small molecules, most likely phosphorylated amino acids, which interacted with U in the supernatant and were not detected in experiments with dead-autoclaved cells. Our study provides new insights into the influence of temperature and cell viability on U phosphate biomineralization by fungi, and furthermore highlight the potential use of P. simplicissimum KS1 particularly for U bioremediation purposes.Graphical Abstract

Neuron-Recognizable Characteristics of Peptides Recombined Using A Neuronal Binding Domain of Botulinum Neurotoxin

Recombinant peptides were designed using the C-terminal domain (receptor binding domain, RBD) and its subdomain (peptide A2) of a heavy chain of botulinum neurotoxin A-type 1 (BoNT/A1), which can bind to the luminal domain of synaptic vesicle glycoprotein 2C (SV2C-LD). Peptide A2- or RBD-containing recombinant peptides linked to an enhanced green fluorescence protein (EGFP) were prepared by expression in Escherichia coli. A pull-down assay using SV2C-LD-covered resins showed that the recombinant peptides for CDC5328 BoNT/A1, referred to EGFP-A2ʹ and EGFP-RBDʹ, exhibited ≥ 2.0-times stronger binding affinity to SV2C-LD than those for the wild-type BoNT/A1. Using bio-layer interferometry, an equilibrium dissociation rate constant (KD) of EGFP-RBDʹ to SV2C-LD was determined to be 5.45 mM, which is 33.87- and 15.67-times smaller than the KD values for EGFP and EGFP-A2ʹ, respectively. Based on confocal laser fluorescence micrometric analysis, the adsorption/absorption of EGFP-RBDʹ to/in differentiated PC-12 cells was 2.49- and 1.29-times faster than those of EGFP and EGFP-A2ʹ, respectively. Consequently, the recombinant peptides acquired reasonable neuron-specific binding/internalizing ability through the recruitment of RBDʹ. In conclusion, RBDs of BoNTs are versatile protein domains that can be used to mark neural systems and treat a range of disorders in neural systems.

Laser Fluorescence and Extinction Methods for Measuring the Flow and Composition of Milk in a Milking Machine

Automation of milking systems is linked to accurate measurement of fluctuations in milk flow during milking. To assess the fluctuations of the milk flow, the formation and movement of milk portions in the milking machine-milk pipeline system was studied. By considering the movement of a milk plug along the milk pipeline, a hydraulic model of the formation of a critical volume of milk in the milking machine manifold was compiled. In practice, the most expedient way of determining milk flow parameters may be to measure the laser fluorescent and extinction responses of moving air-milk mixture. We have implemented a new laser sensing method for measuring the flow rate and composition of milk on the basis of counting the optical response pulses received from moving dispersed components by a CCD array or a randomized fiber optic bundle. Using the developed laser sensors, the theoretical model of milk flow was tested.

Systematic review and meta-analysis of diagnostic studies of proximal surface caries

Aim This systematic review and meta-analysis aimed to assess the diagnostic accuracy and reliability of commonly used caries detection methods for proximal caries diagnostics. Visual examination (VE), bitewing radiography (BWR), laser fluorescence (LF), and fibre-optic transillumination (FOTI) were considered in detail. Material and methods PRISMA guidelines for the reporting of systematic reviews and meta-analyses were applied. The mnemonic PIRDS (problem, index test, reference test, diagnostic and study type) concept was used to guide the literature search. Next, studies that met the inclusion criteria were stepwise selected and evaluated for their quality with a risk of bias (RoB) assessment tool. Studies with low/moderate bias and sufficient reporting were considered for meta-analysis. The pooled sensitivity (SE), specificity (SP), diagnostic odds ratio (DOR), and area under the ROC curve (AUC) were calculated. Results From 129 studies meeting the selection criteria, 31 in vitro studies and five clinical studies were finally included in the meta-analysis. The AUC values for in vitro VE amounted to 0.84 (caries detection) and 0.85 (dentin caries detection). BWR ranged in vitro from 0.55 to 0.82 (caries detection) and 0.81–0.92 (dentin caries detection). LF showed higher AUC values for overall caries detection (0.91) and dentin caries detection (0.83) than did other methods. Clinical data are limited. Conclusion The number of diagnostic studies with low/moderate RoB was found to be low and indicates a need for high-quality, well-designed caries diagnostic studies. Clinical relevance BWR and LF showed good diagnostic performance on proximal surfaces. However, because of the low number of includable clinical studies, these data should be interpreted with caution.

Efficacy of sealants and bonding materials during fixed orthodontic treatment to prevent enamel demineralization: a systematic review and meta-analysis

To analyse clinical studies investigating coating agents such as sealants and other bonding materials to prevent the initiation or inhibit the progress of white spot lesions (WSL) during orthodontic treatment with fixed appliances. Electronic databases (Pubmed, CENTRAL, EMBASE) were screened for studies. No language restrictions were applied. Study selection, data extraction and quality assessment were done in duplicate. Primary outcome included assessment of WSL with visual-tactile assessment and/or laser fluorescence measurements. Twenty-four studies with 1117 patients (age: 11–40 years) and 12,809 teeth were included. Overall, 34 different sealants or bonding materials were analysed. Fourteen studies analysed fluoride and 14 studies non-fluoride releasing materials. Meta-analysis for visual tactile assessment revealed that sealants significantly decreased the initiation of WSL compared to untreated control (RR [95%CI] = 0.70 [0.53; 0.93]; very low level of evidence). Materials releasing fluoride did not decrease initiation of WSL compared to those with no fluoride release (RR [95%CI] = 0.84 [0.70; 1.01]; very low level of evidence). For laser fluorescence measurements no meta-analysis could be performed. The use of sealants seems to be effective in preventing the initiation of post-orthodontic WSL. Furthermore, there is no evidence supporting that fluoride-releasing sealants or bonding materials are more effective than those without fluoride release. No gold standard prevention strategy to prevent WSL during treatment with fixed orthodontic appliances has been established yet. However, based on only a limited number of studies the use of sealants seems to be effective in preventing the initiation of post-orthodontic WSL.