Journal: bioRxiv

Article Title: Cannabis-enriched oral Actinomyces induces anxiety-like behavior via impairing mitochondria and GABA signaling

doi: 10.1101/2025.11.21.689724

Figure Lengend Snippet: The oral swab samples were collected from the mice after chronic oral inoculation of the microbiome, while feces were collected from the intestines at the time of mouse sacrifice (fresh catch). The quantification of A. meyeri, A. odontolyticus , and N. elongata specific DNA was evaluated using qPCR with their specific primers. To count colony-forming units and construct a standard curve for each bacterium, genomic DNA was isolated from four serial dilutions of pure culture through the same DNA extraction method (starting from 1 × 10 9 ). This genomic DNA was then used in the qPCR assay, and the Ct values obtained from the qPCR were employed to generate a standard curve and determine the CFU/mL titer of respective bacteria in mouse oral samples. Data are represented as mean±SEM (n=6 in each group), **p < 0.001, One-way ANOVA, followed by Mann-Whitney U-test.

Article Snippet: Total microbial DNA was extracted from the oral samples using a QIAmp DNA Microbiome kit (Qiagen) according to the manufacturer’s protocol. qPCR was performed using a CFX96 Real-time system (Bio-Rad).

Techniques: Construct, Isolation, DNA Extraction, Bacteria, MANN-WHITNEY

Journal: bioRxiv

Article Title: Cannabis-enriched oral Actinomyces induces anxiety-like behavior via impairing mitochondria and GABA signaling

doi: 10.1101/2025.11.21.689724

Figure Lengend Snippet: (a) Baseline levels before starting the battery of behavioral tests. Home cage locomotor activity was increased in all microbiome treatments as compared to the control (F (3,20) =10.12, p=0.0003). All mice healthily gained their body weight during oral treatments of A. meyeri, A. odontolyticus , and N. elongata . The mice did not show any changes in their serum corticosterone levels, indicating no handling or environmental stress. (b) Y-maze activity showed no change in the spontaneous alternation in any of the microbiome groups as compared to the vehicle control. (c) Different parameters were tested in the Barnes maze on the recall day after extensive five-day training. No significant change was observed in any parameter in any of the microbiome groups. (d) The forced swim test showed a trending increase in immobility time in the A. meyeri treatment, but it was not significant. Data are represented as mean±SEM (n=6 in each group), **p < 0.01, ***p <0.001; One-way ANOVA, followed by Tukey’s multiple comparison test.

Article Snippet: Total microbial DNA was extracted from the oral samples using a QIAmp DNA Microbiome kit (Qiagen) according to the manufacturer’s protocol. qPCR was performed using a CFX96 Real-time system (Bio-Rad).

Techniques: Battery, Activity Assay, Control, Comparison

Journal: bioRxiv

Article Title: Cannabis-enriched oral Actinomyces induces anxiety-like behavior via impairing mitochondria and GABA signaling

doi: 10.1101/2025.11.21.689724

Figure Lengend Snippet: (a) The single cells obtained from each integrated sample, and their UMAP plots, labeled with the cell types, showing that more excitatory neurons were detected and colored by condition. (b) The UMAP plots show the integrated dataset split by individual treatment as compared to the other microbiome groups. Twenty-two distinct cell types were identified through single-cell RNA sequencing, shown in different colors. (c) Gene ontology analysis of KEGG pathways representing the up-(pink) and down-regulated (blue) pathways, gene count (circle size), and their log10 p values in each dataset from different comparisons. (d) Gene ontology analysis based on Reactome/biological processes representing up- and down-regulated processes, -log(10) p-values (color coded), gene count (circle size), and their enrichment scores [-log10(geometric mean of p-values] through the DAVID bioinformatics functional annotation tool. (f) Heatmaps representing the up- and downregulated differentially expressed genes (according to the average log2 fold-change) in different cell types in each integrated dataset.

Article Snippet: Total microbial DNA was extracted from the oral samples using a QIAmp DNA Microbiome kit (Qiagen) according to the manufacturer’s protocol. qPCR was performed using a CFX96 Real-time system (Bio-Rad).

Techniques: Labeling, RNA Sequencing, Functional Assay

Journal: bioRxiv

Article Title: Cannabis-enriched oral Actinomyces induces anxiety-like behavior via impairing mitochondria and GABA signaling

doi: 10.1101/2025.11.21.689724

Figure Lengend Snippet: (A) Schematic illustrating the downregulation of genes associated with GABAergic neurotransmission. Reduced GABA synthesis was indicated by GAD1/GAD2 downregulation; impaired reuptake and recycling by reduced SLC6A1 (GAT1); diminished presynaptic release by CACNA1A downregulation; and weakened postsynaptic signaling by decreased expression of GABRA2, PRKCG, and PRKCA. (B) Quantification of GABA levels in mouse brain homogenates, after normalizing with the total protein content in each sample, revealed significant reductions in A. meyeri –treated mice compared with other microbiome groups. (C) UMAP plots (between-group comparison with A. meyeri group as reference) identified three downregulated genes in GABAergic neurons (Lamp5) and astrocytes: GAD1 (glutamate decarboxylase 67), GAD2 (glutamate decarboxylase 65), and SLC6A1 (GAT1). (D–E) Western blot validation demonstrated decreased GAD1 and GAD2 protein levels, with a trend toward reduced GAT1 expression in both Actinomyces groups relative to vehicle controls. Data are presented as mean ± SEM (n = 3–6 per group). *p < 0.05, **p < 0.01, ***p < 0.001, *** p < 0.00001; one-way ANOVA with Tukey’s post hoc test .

Article Snippet: Total microbial DNA was extracted from the oral samples using a QIAmp DNA Microbiome kit (Qiagen) according to the manufacturer’s protocol. qPCR was performed using a CFX96 Real-time system (Bio-Rad).

Techniques: Expressing, Comparison, Western Blot, Biomarker Discovery

Journal: Journal of Clinical Medicine

Article Title: Does Postoperative Oral and Intestinal Microbiota Correlate with the Weight-Loss Following Bariatric Surgery?—A Cohort Study

doi: 10.3390/jcm9123863

Figure Lengend Snippet: Differences in oral microbiota between Group 1 and Group 2 (1—Group 1; 0—Group 2).

Article Snippet: To increase the accuracy and decrease the risk of bias, three negative controls and two positive controls and ATCC standards for oral microbiome (ATCC ® MSA-1004TM) and gut microbiome (ATCC ® MSA-1006TM, Manassas, Virginia, United States) were included [ ].

Techniques:

Journal: Journal of Clinical Medicine

Article Title: Does Postoperative Oral and Intestinal Microbiota Correlate with the Weight-Loss Following Bariatric Surgery?—A Cohort Study

doi: 10.3390/jcm9123863

Figure Lengend Snippet: Differences in oral microbiota between Group 1 and Group 2 (1—Group 1; 0—Group 2).

Article Snippet: To increase the accuracy and decrease the risk of bias, three negative controls and two positive controls and ATCC standards for oral microbiome (ATCC ® MSA-1004TM) and gut microbiome (ATCC ® MSA-1006TM, Manassas, Virginia, United States) were included [ ].

Techniques: