Investigating Oral Microbiome Profiles in Children with Cleft Lip and Palate for Prognosis of Alveolar Bone Grafting

<div><p>In this study, we sought to investigate the oral microbiota structure of children with cleft lip and palate (CLP) and explore the pre-operative oral bacterial composition related to the prognosis of alveolar bone grafting. In total, 28 patients (19 boys, 9 girls) with CLP who were scheduled to undergo alveolar bone grafting for the first time were recruited. According to the clinical examination of operative sites at the third month after the operation, the individuals were divided into a non-inflammation group (n = 15) and an inflammation group (n = 13). In all, 56 unstimulated saliva samples were collected before and after the operation. The v3-v4 hypervariable regions of the <i>16S rRNA</i> gene were sequenced using an Illumina MiSeq sequencing platform. Based on the beta diversity of the operational taxonomic units (OTUs) in the inflammation and non-inflammation samples, the microbial variation in the oral cavity differed significantly between the two groups before and after the operation (P < 0.05). Analysis of the relative abundances of pre-operative OTUs revealed 26 OTUs with a relative abundance higher than 0.01%, reflecting a significant difference of the relative abundance between groups (P < 0.05). According to a principal component analysis of the pre-operative samples, the inflammation-related OTUs included <i>Tannerella sp</i>., <i>Porphyromonas sp</i>., <i>Gemella sp</i>., <i>Moraxella sp</i>., <i>Prevotella nigrescens</i>, and <i>Prevotella intermedia</i>, most of which were enriched in the inflammation group and showed a significant positive correlation. A cross-validated random forest model based on the 26 different OTUs before the operation was able to fit the post-operative status of grafted sites and yielded a good classification result. The sensitivity and specificity of this classified model were 76.9% and 86.7%, respectively. These findings show that the oral microbiota profile before alveolar bone grafting may be related to the risk of post-operative inflammation at grafted sites.</p></div

Principal component analysis (PCA) of the pre-operative OTUs information for the post-operative inflammation and non-inflammation groups.

<p>Based on 26 different OTUs of the pre-operation (relative abundance higher than 0.01%) between the two groups, the inflammation samples appeared to cluster together in the direction opposite to that of the non-inflammation samples. The 14 OTUs with the pink shadow on the right of the diagram were closely related to post-operative inflammation at the grafted sites.</p

OTUs differing between the non-inflammation and inflammation groups before and after the operation.

<p>In total, 26 OTUs with relative abundances higher than 0.01% exhibited significant differences in mean relative abundances between the non-inflammation and inflammation groups before the operation (Mann-Whitney test, P < 0.05). Bars indicate means ± SEM relative abundances. The upper part of the diagram shows that the levels of 13 pre-operative OTUs were higher in the inflammation group, and the lower part shows that another 13 OTUs were lower. The relative abundance of four OTUs marked with the stars (★) were significantly different between the inflammation and non-inflammation groups both pre- and post-operation.A higher proportion of OTU4340587, corresponding to <i>Porphyromonas sp</i>. (with the pink shadow), was found in inflammation subjects than in non-inflammation subjects both pre- and post-operation.</p

Pre-operative OTUs enriched in saliva samples of inflammation and non-inflammation subjects.

<p>Co-occurrence networks were constructed based on the relative abundance profiles of pre-operative OTUs differing between the inflammation and non-inflammation groups. Each node represents a different OTU, and the most abundant OTUs (relative abundance higher than 0.01%) are colored and annotated to indicate the species or unclassified species in a genus (sp.). The sizes of the nodes are proportional to the average relative abundances of the OTUs. Blue and red edges between each pair of OTUs indicate significantly positive and negative correlations, respectively (Spearman’s correlation coefficient > 0.4 or < -0.4, P < 0.05).</p

Beta diversity of the oral microbiome of non-inflammation and inflammation groups based on the unweighted UniFrac metric.

<p>(A) The inflammation samples were significantly more similar to one another than the non-inflammation samples both in the pre- and post-operative comparison. Principal coordinate analysis (PCoA) plot shows clustering of most inflammation samples, which were separated from non-inflammation samples (B) before and (C) after the operation. Red and green dots indicate inflammation and non-inflammation samples, respectively. * 0.01 < P < 0.05, ** P < 0.01.</p

Dehmographic and clinical characteristics of studied subjects.

<p>Dehmographic and clinical characteristics of studied subjects.</p

Design, Synthesis, and Biological Evaluation of Novel Chromanone Derivatives as Multifunctional Agents for the Treatment of Alzheimer’s Disease

Based on a multitarget strategy, a series of novel chromanone–1-benzyl-1,2,3,6-tetrahydropyridin hybrids were identified for the potential treatment of Alzheimer’s disease (AD). Biological evaluation demonstrated that these hybrids exhibited significant inhibitory activities toward acetylcholinesterase (AChE) and monoamine oxidase B (MAO-B). The optimal compound C10 possessed excellent dual AChE/MAO-B inhibition both in terms of potency and equilibrium (AChE: IC50 = 0.58 ± 0.05 μM; MAO-B: IC50 = 0.41 ± 0.04 μM). Further molecular modeling and kinetic investigations revealed that compound C10 was a dual-binding inhibitor bound to both the catalytic anionic site and peripheral anionic site of AChE. In addition, compound C10 exhibited low neurotoxicity and potently inhibited AChE enzymatic activity. Furthermore, compound C10 more effectively protected against mitochondrial dysfunction and oxidation than donepezil, strongly inhibited AChE-induced amyloid aggregation, and moderately reduced glutaraldehyde-induced phosphorylation of tau protein in SH-SY5Y cells. Moreover, compound C10 displayed largely enhanced improvements in cognitive behaviors and spatial memory in a scopolamine-induced AD mice model with better efficacy than donepezil. Overall, the multifunctional profiles of compound C10 suggest that it deserves further investigation as a promising lead for the prospective treatment of AD