Efficacy of the TuttleNumbNow Intraosseous Method for Pulpal Anesthesia in the Mandibular First Molar: A Prospective, Randomized, Crossover Study.

INTRODUCTION: Previous studies on intraosseous (IO) anesthesia as a primary injection have shown high success rates. The TuttleNumbNow (TNN; Orem, UT) is a new primary IO injection technique that has not been scientifically evaluated. Therefore, the purpose of this prospective randomized, crossover study was to evaluate the anesthetic efficacy of the TNN IO technique using the Septoject Evolution needle (Septodont, Saint-Maur-des-Fosses, France) compared with buccal infiltration for pulpal anesthesia in mandibular first molars. METHODS: One hundred four healthy subjects were randomly assigned to 2 treatment groups separated by at least 2 weeks. One set of injections consisted of buccal infiltration of the mandibular first molar using 1.8 mL 4% articaine with 1:100,000 epinephrine followed by a mock TNN injection distal to the mandibular first molar. The other set of injections was a mock buccal infiltration of the mandibular first molar followed by a TNN injection of 1.8 mL 4% articaine with 1:100,000 epinephrine distal to the mandibular first molar. Statistical analyses were performed. RESULTS: For the mandibular first molar, which had a 42% anesthetic success rate (highest 80 reading) with buccal infiltration compared with 49% with the TNN, no statistically significant difference in success was observed (P = .2115). CONCLUSIONS: The TNN technique has been advocated as an IO injection. However, the inability to deliver anesthetic solution to the cancellous bone resulted in an anesthetic success rate of 49%. The success was statistically similar to a buccal infiltration (42%) and would not provide adequate pulpal anesthesia as a primary injection.

Effect of Intrapapillary Local Anesthetic on Postoperative Pain Following Dental Treatment Under General Anesthesia in Pediatric Patients.

OBJECTIVE: This randomized, prospective, blinded study compared pain in children following dental treatment under general anesthesia (GA) using 1 of 2 established analgesia methods. METHODS: Patients age 4 to 7 years were randomly assigned to a control group (intravenous [IV] analgesics) or experimental group (IV analgesics and intrapapillary local anesthetic infiltrations) between July 2017 and February 2018. During recovery from surgery, Faces, Legs, Activity, Cry, and Consolability (FLACC) scores were recorded upon regaining consciousness and reassessed every 15 minutes until discharge. Overall pain occurrence (FLACC ≥1) and moderate/severe pain occurrence (FLACC ≥4) were analyzed using mixed effects logistic regression (N = 88). RESULTS: The experimental group had a 17% lower overall pain occurrence than the control group (16 vs 33%; p = .02). Moderate/severe pain occurrence between the groups was not significant (9 vs 22%; p = .23). The dental treatment subjects received (number of completed stainless steel crowns, extractions, and/or pulpotomies) did not significantly affect pain occurrence. CONCLUSION: Local anesthesia intrapapillary infiltrations around stainless steel crowns decrease overall pain occurrence but not moderate/severe pain occurrence following dental treatment under GA in pediatric patients.

Nutrition and Frailty: Opportunities for Prevention and Treatment.

Frailty is a syndrome of growing importance given the global ageing population. While frailty is a multifactorial process, poor nutritional status is considered a key contributor to its pathophysiology. As nutrition is a modifiable risk factor for frailty, strategies to prevent and treat frailty should consider dietary change. Observational evidence linking nutrition with frailty appears most robust for dietary quality: for example, dietary patterns such as the Mediterranean diet appear to be protective. In addition, research on specific foods, such as a higher consumption of fruit and vegetables and lower consumption of ultra-processed foods are consistent, with healthier profiles linked to lower frailty risk. Few dietary intervention studies have been conducted to date, although a growing number of trials that combine supplementation with exercise training suggest a multi-domain approach may be more effective. This review is based on an interdisciplinary workshop, held in November 2020, and synthesises current understanding of dietary influences on frailty, focusing on opportunities for prevention and treatment. Longer term prospective studies and well-designed trials are needed to determine the causal effects of nutrition on frailty risk and progression and how dietary change can be used to prevent and/or treat frailty in the future.

Calystegines are Potential Urine Biomarkers for Dietary Exposure to Potato Products.

SCOPE: Metabolites derived from specific foods present in urine samples can provide objective biomarkers of food intake (BFIs). This study investigated the possibility that calystegines (a class of iminosugars) may provide BIFs for potato (Solanum tuberosum L.) product exposure. METHODS AND RESULTS: Calystegine content is examined in published data covering a wide range of potato cultivars. Rapid methods are developed for the quantification of calystegines in cooked potato products and human urine using triple quadrupole mass spectrometry. The potential of calystegines as BFIs for potato consumption is assessed in a controlled food intervention study in the United Kingdom and validated in an epidemiological study in Portugal. Calystegine concentrations are reproducibly above the quantification limit in first morning void urines the day after potato consumption, showing a good dose-response relationship, particularly for calystegine A3 . The design of the controlled intervention mimicks exposure to a typical UK diet and showed that neither differences in preparation/cooking method or influence of other foods in the diet has significant impact on biomarker performance. Calystegine biomarkers also perform well in the independent validation study. CONCLUSION: It is concluded that calystegines have many of the characteristics needed to be considered as specific BFIs for potato product intake.

Integrated Analytical and Statistical Two-Dimensional Spectroscopy Strategy for Metabolite Identification: Application to Dietary Biomarkers.

A major purpose of exploratory metabolic profiling is for the identification of molecular species that are statistically associated with specific biological or medical outcomes; unfortunately, the structure elucidation process of unknowns is often a major bottleneck in this process. We present here new holistic strategies that combine different statistical spectroscopic and analytical techniques to improve and simplify the process of metabolite identification. We exemplify these strategies using study data collected as part of a dietary intervention to improve health and which elicits a relatively subtle suite of changes from complex molecular profiles. We identify three new dietary biomarkers related to the consumption of peas (N-methyl nicotinic acid), apples (rhamnitol), and onions (N-acetyl-S-(1Z)-propenyl-cysteine-sulfoxide) that can be used to enhance dietary assessment and assess adherence to diet. As part of the strategy, we introduce a new probabilistic statistical spectroscopy tool, RED-STORM (Resolution EnhanceD SubseT Optimization by Reference Matching), that uses 2D J-resolved 1H NMR spectra for enhanced information recovery using the Bayesian paradigm to extract a subset of spectra with similar spectral signatures to a reference. RED-STORM provided new information for subsequent experiments (e.g., 2D-NMR spectroscopy, solid-phase extraction, liquid chromatography prefaced mass spectrometry) used to ultimately identify an unknown compound. In summary, we illustrate the benefit of acquiring J-resolved experiments alongside conventional 1D 1H NMR as part of routine metabolic profiling in large data sets and show that application of complementary statistical and analytical techniques for the identification of unknown metabolites can be used to save valuable time and resources.

Diversity and association of phenotypic and metabolomic traits in the close model grasses Brachypodium distachyon, B. stacei and B. hybridum.

Background and Aims: Morphological traits in combination with metabolite fingerprinting were used to investigate inter- and intraspecies diversity within the model annual grasses Brachypodium distachyon, Brachypodium stacei and Brachypodium hybridum . Methods: Phenotypic variation of 15 morphological characters and 2219 nominal mass ( m / z ) signals generated using flow infusion electrospray ionization-mass spectrometry (FIE-MS) were evaluated in individuals from a total of 174 wild populations and six inbred lines, and 12 lines, of the three species, respectively. Basic statistics and multivariate principal component analysis and discriminant analysis were used to differentiate inter- and intraspecific variability of the two types of variable, and their association was assayed with the rcorr function. Key Results: Basic statistics and analysis of variance detected eight phenotypic characters [(stomata) leaf guard cell length, pollen grain length, (plant) height, second leaf width, inflorescence length, number of spikelets per inflorescence, lemma length, awn length] and 434 tentatively annotated metabolite signals that significantly discriminated the three species. Three phenotypic traits (pollen grain length, spikelet length, number of flowers per inflorescence) might be genetically fixed. The three species showed different metabolomic profiles. Discriminant analysis significantly discriminated the three taxa with both morphometric and metabolome traits and the intraspecific phenotypic diversity within B. distachyon and B. stacei . The populations of B. hybridum were considerably less differentiated. Conclusions: Highly explanatory metabolite signals together with morphological characters revealed concordant patterns of differentiation of the three taxa. Intraspecific phenotypic diversity was observed between northern and southern Iberian populations of B. distachyon and between eastern Mediterranean/south-western Asian and western Mediterranean populations of B. stacei . Significant association was found for pollen grain length and lemma length and ten and six metabolomic signals, respectively. These results would guide the selection of new germplasm lines of the three model grasses in ongoing genome-wide association studies.

High-throughput, nontargeted metabolite fingerprinting using nominal mass flow injection electrospray mass spectrometry.

Flow injection electrospray-mass spectrometry (FIE-MS) is finding utility as a first-pass metabolite fingerprinting tool in many research fields. We provide a protocol that has proved reliable in large-scale research projects involving diverse sample matrices originating from plants, microbes and mammalian biofluids. Using Brachypodium leaf material as an example matrix all steps are summarized from sample extraction to data quality assessment. Alternative procedures for dealing with other common matrices such as bloods and urine are included. With little sample pretreatment, no chromatography and instrument cycle times of <5 min it is feasible to analyze >1,000 samples per week. Analysis of a typical batch of 240 samples (including first-pass data analysis) can be accomplished within 48 h.

Representation, comparison, and interpretation of metabolome fingerprint data for total composition analysis and quality trait investigation in potato cultivars.

Understanding attributes of crop varieties and food raw materials underlying desirable characteristics is a significant challenge. Metabolomics technology based on flow infusion electrospray ionization mass spectrometry (FIE-MS) has been used to investigate the chemical composition of potato cultivars associated with quality traits in harvested tubers. Through the combination of metabolite fingerprinting with random forest data modeling, a subset of metabolome signals explanatory of compositional differences between individual genotypes were ranked for importance. Interpretative analysis of highlighted signals based on ranking behavior, intensity correlations, and mathematical relationships of ion masses correctly predicted metabolites associated with flavor and pigmentation traits in potato tubers. GC-MS profiling was used to further validate proposed compositional differences. The potential for the development of a database strategy for large scale, long-term projects requiring comparison of chemical composition in plant breeding, mutant population analysis in functional genomics experiments, or food raw material analysis is described.

Hierarchical metabolomics demonstrates substantial compositional similarity between genetically modified and conventional potato crops.

There is current debate whether genetically modified (GM) plants might contain unexpected, potentially undesirable changes in overall metabolite composition. However, appropriate analytical technology and acceptable metrics of compositional similarity require development. We describe a comprehensive comparison of total metabolites in field-grown GM and conventional potato tubers using a hierarchical approach initiating with rapid metabolome "fingerprinting" to guide more detailed profiling of metabolites where significant differences are suspected. Central to this strategy are data analysis procedures able to generate validated, reproducible metrics of comparison from complex metabolome data. We show that, apart from targeted changes, these GM potatoes in this study appear substantially equivalent to traditional cultivars.

Comparison of rapid liquid chromatography-electrospray ionization-tandem mass spectrometry methods for determination of glycoalkaloids in transgenic field-grown potatoes.

Two rapid methods for highly selective detection and quantification of the two major glycoalkaloids in potatoes, alpha-chaconine and alpha-solanine, were compared for robustness in high-throughput operations for over 1000 analytical runs using potato tuber samples from field trials. Glycoalkaloids were analyzed using liquid chromatography coupled to tandem mass spectrometry in multiple reaction monitoring mode. An electrospray interface was used in the detection of glycoalkaloids in positive ion mode. Classical reversed phase (RP) and hydrophilic interaction (HILIC) columns were investigated for chromatographic separation, ruggedness, recovery, precision, and accuracy. During the validation procedure both methods proved to be precise and accurate enough in relation to the high degree of endogenous biological variability found for field-grown potato tubers. However, the RP method was found to be more precise, more accurate, and, more importantly, more rugged than the HILIC method for maintaining the analytes' peak shape symmetry in high-throughput operation. When applied to the comparison of six classically bred potato cultivars to six genetically modified (GM) lines engineered to synthesize health beneficial inulins, the glycoalkaloid content in potato peels of all GM lines was found within the range of the six cultivars. We suggest complementing current unbiased metabolomic strategies by validating quantitative analytical methods for important target analytes such as the toxic glycoalkaloids in potato plants.