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Background: Malondialdehyde–acetaldehyde (MAA) adducts lead to generation of anti‐MAA autoantibodies and have been independently identified in inflamed periodontal and rheumatoid arthritis (RA) tissues. This study evaluates serum samples from RA cases and osteoarthritis (OA) controls to quantify associations between periodontal clinical measures, alveolar bone loss (ABL), and anti‐Porphyromonas gingivalis, anti‐Prevotella intermedia, and anti‐Fusobacterium nucleatum antibody concentrations with anti‐MAA antibody concentrations. Methods: Participants (n = 284 RA cases, n = 330 OA controls) underwent periodontal clinical assessments and ABL measurements. Serum immunoglobulin (Ig) A, IgG, and IgM anti‐MAA and serum IgG antibacterial antibody concentrations were quantified by enzyme‐linked immunosorbent assay (ELISA). Analyses utilized simple linear regression and multivariable adjusted models. Results: No significant associations of periodontal clinical measures with serum anti‐MAA were found. Moderate (p = 0.038 and p = 0.036, respectively) and high ABL (p = 0.012 and p = 0.014, respectively) in RA cases (but not in OA) were positively associated with IgG and IgM anti‐MAA. Anti‐P. gingivalis and anti‐P. intermedia antibody concentrations were positively associated with IgA (p = 0.001 for both), IgG (p = 0.007 and p = 0.034, respectively), and IgM anti‐MAA antibody concentrations (p < 0.001 and p = 0.020, respectively), while anti‐F. nucleatum was positively associated with IgG anti‐MAA (p = 0.042), findings that were similar across groups. Conclusions: A positive association was demonstrated between ABL and serum IgG and IgM anti‐MAA antibody concentrations that was unique to RA and not observed in OA. Serum anti‐P. gingivalis, anti‐P. intermedia, and anti‐F. nucleatum antibody concentrations displayed significant associations with anti‐MAA antibody in both groups. These findings suggest MAA may play a role in the interrelationship between the periodontium and RA. [ABSTRACT FROM AUTHOR]

Objectives: In the present study, we aimed to clarify the mechanisms by which periodontal pathogens, particularly Prevotella intermedia, induce severe neutrophilic inflammation. In addition, we aimed to test the efficacy of macrolides, which has not been resolved in the neutrophilic inflammation induced by P. intermedia. Methods: NCl-H292 human airway epithelial cells were pre-incubated with clarithromycin for 2 h before incubation with P. intermedia supernatants. Then, C-X-C motif chemokine ligand 8 (CXCL8) transcription and interleukin (IL)-8 production were measured. To elucidate the signaling pathway, mitogen-activated protein kinase inhibitors were added to the cell culture, and the cells were subjected to Western blotting. Results:P. intermedia supernatants promoted CXCL8 transcription and IL-8 production, and the reactions were significantly suppressed by clarithromycin pretreatment. Only trametinib, the selective mitogen-activated extracellular signal-regulated kinase inhibitor, downregulated CXCL8 transcription and IL-8 production. Furthermore, Western blotting revealed that stimulation with P. intermedia supernatants specifically induces extracellular signal-regulated kinases (ERK) 1/2 phosphorylation, which is suppressed by clarithromycin pretreatment. Notably, the interference analysis revealed that ERK3 might be dispensable for IL-8 production under the stimulation of P. intermedia supernatants. Conclusions: Our results provide new insight into the mechanism underlying P. intermedia-induced production of IL-8 from human airway epithelial cells. Furthermore, macrolides might have therapeutic potential in regulating periodontal pathogen-induced neutrophilic inflammation in the lungs. [ABSTRACT FROM AUTHOR]

AbstractPrevotella intermedia is a Gram-negative anaerobic bacterium that is a common pathogen of periodontitis. Recent studies have revealed that P. intermedia is closely associated with a variety of diseases involving multiple systems. Under the action of its virulence factors such as cysteine protease and adhesins, P. intermedia has the ability to bind and invade various host cells including gingival fibroblasts. It can also copolymerize a variety of pathogenic bacteria, leading to interference with the host’s immune inflammatory response and causing various diseases. In this article, we review the progress of research on P. intermedia virulence factors and bacterial pathogenesis, and the correlation between P. intermedia and various diseases. [ABSTRACT FROM AUTHOR]

Abstract Background Oral microbial dysbiosis contributes to the development of oral squamous cell carcinoma (OSCC). Our previous study showed that Prevotella intermedia (P. intermedia) were enriched in the oral mucosal surface, plaque, and saliva of patients with OSCC. Intratumoral microbiome could reshape the immune system and influence the development of various tumors. However, the invasion status of human OSCC tissues by P. intermedia and the pathway through which intratumoral P. intermedia potentiates tumor progression remain unexplored. Methods P. intermedia in human OSCC or normal tissues was detected by FISH. A mouse OSCC cell line SCC7 was adopted to investigate the effects of heat-killed P. intermedia treatment on cell proliferation, invasion, and cytokine release by using CCK-8 assay, transwell invasion assay and ELISA. Moreover, we established a mouse transplanted tumor model by using SCC7 cells, injected heat-killed P. intermedia into tumor tissues, and investigated the effects of heat-killed P. intermedia on tumor growth, invasion, cytokine levels, immune cell infiltrations, and expression levels by using gross observation, H&E staining, ELISA, immunohistochemistry, mRNA sequencing, and transcriptomic analysis. Results Our results indicated that P. intermedia were abundant in OSCC and surrounding muscle tissues. Heat-killed P. intermedia promoted SCC7 cell proliferation, invasion and proinflammatory cytokine secretions, accelerated transplanted tumor growth in mice, exacerbate muscle and perineural invasion of OSCC, elevated the serum levels of IL-17A, IL-6, TNF-α, IFN-γ, and PD-L1, induced Treg cells M2 type macrophages in mouse transplanted tumors. The data of transcriptomic analysis revealed that heat-killed P. intermedia increased the expression levels of inflammatory cytokines and chemokines while reduced the expression levels of some tumor suppressor genes in mouse transplanted tumors. Additionally, IL-17 signaling pathway was upregulated whereas GABAergic system was downregulated by heat-killed P. intermedia treatment. Conclusions Taken together, our results suggest that P. intermedia could inhibit the expression of tumor suppressors, alter the tumor microenvironment, and promote the progression of OSCC.

Purpose: To investigate the effects of periopathogens on bone grafts for maxillary sinus floor augmentation and implant survival. Materials and Methods: A cohort study was designed. Samples from maxillary sinus floor augmentation sites and from periodontal pockets in adjacent teeth of the same patients were collected during the surgery and following periodontal maintenance and plaque removal. Polymerase chain reaction (PCR) was performed by an external lab service for analysis of the following periopathogens: Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythia, and Treponema denticola. Correlations between the presence of periopathogens in sinus sites and periodontal pockets in adjacent teeth and the clinical findings (bone and implant survival) were studied. Results: A total of 23 patients were suitable for study inclusion. Periopathogens were found in sinus sites in 6 patients (26%) and in tooth sites in 15 patients (65%). There was low correlation for the presence of periopathogens between tooth sites and sinus sites and for complications. Conclusion: The presence of periopathogens as revealed by PCR was lower in sinus sites compared with tooth sites. Their presence neither caused bone pathology nor affected dental implant survival. The presence of periopathogens in sinus sites is inevitable even under periodontal maintenance. Although its presence did not endanger implant survival, future studies are recommended for studying this issue. Further analyses of the presence of periopathogens in cases of bone resorption at the apex of dental implants inserted into sites of augmented sinuses are warranted. [ABSTRACT FROM AUTHOR]

Purpose: Periodontitis-associated bacteria, such as Porphyromonas gingivalis and Fusobacterium nucleatum, are closely linked to the risk of oral squamous cell carcinoma (OSCC). Emerging studies have indicated that another common periodontal pathogen, Prevotella intermedia (P. intermedia), is enriched in OSCC and could affect the occurrence and progression of OSCC. Our aim is to determine the effects of P. intermedia on the progression of OSCC and the role of antibiotics in reversing these effects. Methods: In this study, a murine xenograft model of OSCC was established, and the mice were injected intratumorally with PBS (control group), P. intermedia (P.i group), or P. intermedia combined with an antibiotic cocktail administration (P.i + ABX group), respectively. The effects of P. intermedia and ABX administration on xenograft tumor growth, invasion, angiogenesis, and metastasis were investigated by tumor volume measurement and histopathological examination. Enzyme-linked immunosorbent assay (ELISA) was used to investigate the changes in serum cytokine levels. Immunohistochemistry (IHC) was adopted to analyze the alterations in the levels of inflammatory cytokines and infiltrated immune cells in OSCC tissues of xenograft tumors. Transcriptome sequencing and analysis were conducted to determine differential expression genes among various groups. Results: Compared with the control treatment, P. intermedia treatment significantly promoted tumor growth, invasion, angiogenesis, and metastasis, markedly affected the levels of inflammatory cytokines, and markedly altered M2 macrophages and regulatory T cells (Tregs) infiltration in the tumor microenvironment. However, ABX administration clearly abolished these effects of P. intermedia. Transcriptome and immunohistochemical analyses revealed that P. intermedia infection increased the expression of interferon-stimulated gene 15 (ISG15). Correlation analysis indicated that the expression level of ISG15 was positively correlated with the Ki67 expression level, microvessel density, serum concentrations and tissue expression levels of inflammatory cytokines, and quantities of infiltrated M2 macrophages and Tregs. However, it is negatively correlated with the quantities of infiltrated CD4+ and CD8+ T cells. Conclusion: In conclusion, intratumoral P. intermedia infection aggravated OSCC progression, which may be achieved through upregulation of ISG15. This study sheds new light on the possible pathogenic mechanism of intratumoral P. intermedia in OSCC progression, which could be a prospective target for OSCC prevention and treatment. [ABSTRACT FROM AUTHOR]

The indigenous microbial milieu within tumorous tissues exerts a pivotal influence on the genesis and advancement of gastric cancer (GC). This investigation scrutinizes the functions and molecular mechanisms attributed to Prevotella intermedia in the malignant evolution of GC. Isolation of P. intermedia from paired GC tissues was undertaken. Quantification of P. intermedia abundance in 102 tissues was accomplished using quantitative real‐time PCR (qRT‐PCR). Assessment of the biological effects of P. intermedia on GC cells was observed using culture medium supernatant. Furthermore, the protein profile of GC cells treated with tumor‐derived P. intermedia was examined through label‐free protein analysis. The functionality of perilipin 3 (PLIN3) was subsequently confirmed using shRNA. Our investigation revealed that the relative abundance of P. intermedia in tumor tissues significantly surpassed that of corresponding healthy tissues. The abundance of P. intermedia exhibited correlations with tumor differentiation (p = 0.006), perineural invasion (p = 0.004), omentum majus invasion (p = 0.040), and the survival duration of GC patients (p = 0.042). The supernatant derived from tumor‐associated P. intermedia bolstered the proliferation, clone formation, migration, and invasion of GC cells. After indirect co‐cultivation with tumor‐derived P. intermedia, dysregulation of 34 proteins, including PLIN3, was discerned in GC cells. Knockdown of PLIN3 mitigated the malignancy instigated by P. intermedia in GC cells. Our findings posit that P. intermedia from the tumor microenvironment plays a substantial role in the malignant progression of GC via the modulation of PLIN3 expression. Moreover, the relative abundance of P. intermedia might serve as a potential biomarker for the diagnosis and prognosis of GC. [ABSTRACT FROM AUTHOR]

Background: Oral microbial dysbiosis contributes to the development of oral squamous cell carcinoma (OSCC). Our previous study showed that Prevotella intermedia (P. intermedia) were enriched in the oral mucosal surface, plaque, and saliva of patients with OSCC. Intratumoral microbiome could reshape the immune system and influence the development of various tumors. However, the invasion status of human OSCC tissues by P. intermedia and the pathway through which intratumoral P. intermedia potentiates tumor progression remain unexplored. Methods: P. intermedia in human OSCC or normal tissues was detected by FISH. A mouse OSCC cell line SCC7 was adopted to investigate the effects of heat-killed P. intermedia treatment on cell proliferation, invasion, and cytokine release by using CCK-8 assay, transwell invasion assay and ELISA. Moreover, we established a mouse transplanted tumor model by using SCC7 cells, injected heat-killed P. intermedia into tumor tissues, and investigated the effects of heat-killed P. intermedia on tumor growth, invasion, cytokine levels, immune cell infiltrations, and expression levels by using gross observation, H&E staining, ELISA, immunohistochemistry, mRNA sequencing, and transcriptomic analysis. Results: Our results indicated that P. intermedia were abundant in OSCC and surrounding muscle tissues. Heat-killed P. intermedia promoted SCC7 cell proliferation, invasion and proinflammatory cytokine secretions, accelerated transplanted tumor growth in mice, exacerbate muscle and perineural invasion of OSCC, elevated the serum levels of IL-17A, IL-6, TNF-α, IFN-γ, and PD-L1, induced Treg cells M2 type macrophages in mouse transplanted tumors. The data of transcriptomic analysis revealed that heat-killed P. intermedia increased the expression levels of inflammatory cytokines and chemokines while reduced the expression levels of some tumor suppressor genes in mouse transplanted tumors. Additionally, IL-17 signaling pathway was upregulated whereas GABAergic system was downregulated by heat-killed P. intermedia treatment. Conclusions: Taken together, our results suggest that P. intermedia could inhibit the expression of tumor suppressors, alter the tumor microenvironment, and promote the progression of OSCC. [ABSTRACT FROM AUTHOR]

Objectives: In the present study, we aimed to clarify the mechanisms by which periodontal pathogens, particularly Prevotella intermedia, induce severe neutrophilic inflammation. In addition, we aimed to test the efficacy of macrolides, which has not been resolved in the neutrophilic inflammation induced by P. intermedia. Methods: NCl-H292 human airway epithelial cells were pre-incubated with clarithromycin for 2 h before incubation with P. intermedia supernatants. Then, C-X-C motif chemokine ligand 8 (CXCL8) transcription and interleukin (IL)-8 production were measured. To elucidate the signaling pathway, mitogen-activated protein kinase inhibitors were added to the cell culture, and the cells were subjected to Western blotting. Results:P. intermedia supernatants promoted CXCL8 transcription and IL-8 production, and the reactions were significantly suppressed by clarithromycin pretreatment. Only trametinib, the selective mitogen-activated extracellular signal-regulated kinase inhibitor, downregulated CXCL8 transcription and IL-8 production. Furthermore, Western blotting revealed that stimulation with P. intermedia supernatants specifically induces extracellular signal-regulated kinases (ERK) 1/2 phosphorylation, which is suppressed by clarithromycin pretreatment. Notably, the interference analysis revealed that ERK3 might be dispensable for IL-8 production under the stimulation of P. intermedia supernatants. Conclusions: Our results provide new insight into the mechanism underlying P. intermedia-induced production of IL-8 from human airway epithelial cells. Furthermore, macrolides might have therapeutic potential in regulating periodontal pathogen-induced neutrophilic inflammation in the lungs.

Nitro-conjugated linoleic acid (NO2-CLA) has been observed to manifest salutary signaling responses, including anti-inflammatory and antioxidant properties. Here, the authors have explored the influence and underlying mechanisms of NO2-CLA on the proinflammatory reaction of murine macrophages that were challenged with lipopolysaccharide (LPS) derived from Prevotella intermedia, a putative periodontopathic bacterium. Treatment of LPS-activated RAW264.7 cells with NO2-CLA notably dampened the secretion of iNOS-derived NO, IL-1β and IL-6 as well as their gene expressions and significantly enhanced the markers for M2 macrophage polarization. NO2-CLA promoted the HO-1 expression in cells challenged with LPS, and tin protoporphyrin IX, an HO-1 inhibitor, significantly reversed the NO2-CLA-mediated attenuation of NO secretion, but not IL-1β or IL-6. We found that cells treated with NO2-CLA significantly increased mRNA expression of PPAR-γ compared to control cells, and NO2-CLA significantly reverted the decrease in PPAR-γ mRNA caused by LPS. Nonetheless, antagonists to PPAR-γ were unable to reverse the NO2-CLA-mediated suppression of inflammatory mediators. In addition, NO2-CLA did not alter the p38 and JNK activation elicited by LPS. Both NF-κB reporter activity and IκB-α degradation caused by LPS were notably diminished by NO2-CLA. NO2-CLA was observed to interrupt the nuclear translocation and DNA binding of p50 subunits caused by LPS with no obvious alterations in p65 subunits. Further, NO2-CLA attenuated the phosphorylation of STAT1/3 elicited in response to LPS. We propose that NO2-CLA could be considered as a possible strategy for the therapy of periodontal disease, although additional researches are certainly required to confirm this. [ABSTRACT FROM AUTHOR]

Abstract Prevotella intermedia, a Gram‐negative bacterium from the Bacteroidota phylum, is associated with periodontitis. Other species within this phylum are known to possess the general O‐glycosylation system. The O‐glycoproteome has been characterized in several species, including Tannerella forsythia, Porphyromonas gingivalis, and Flavobacterium johnsoniae. In our study, we used electron cryotomography (cryoET) and glycoproteomics to reveal the ultrastructure of P. intermedia and characterize its O‐glycoproteome. Our cryoET analysis unveiled the ultrastructural details of the cell envelope and outer membrane vesicles (OMVs) of P. intermedia. We observed an electron‐dense surface layer surrounding both cells and OMVs. The OMVs were often large (>200 nm) and presented two types, with lumens being either electron‐dense or translucent. LC‐MS/MS analyses of P. intermedia fractions led to the identification of 1655 proteins, which included 62 predicted T9SS cargo proteins. Within the glycoproteome, we identified 443 unique O‐glycosylation sites within 224 glycoproteins. Interestingly, the O‐glycosylation motif exhibited a broader range than reported in other species, with O‐glycosylation found at D(S/T)(A/I/L/M/T/V/S/C/G/F/N/E/Q/D/P). We identified a single O‐glycan with a delta mass of 1531.48 Da. Its sequence was determined by MS2 and MS3 analyses using both collision‐induced dissociation and high‐energy collisional dissociation fragmentation modes. After partial deglycosylation with trifluoromethanesulfonic acid, the O‐glycan sequence was confirmed to be dHex‐dHex‐HexNAc (HPO3‐C6H12O5)‐dHex‐Hex‐HexA‐Hex(dHex). Bioinformatic analyses predicted the localization of O‐glycoproteins, with 73 periplasmic proteins, 53 inner membrane proteins, 52 lipoproteins, 26 outer membrane proteins, and 14 proteins secreted by the T9SS.

Abstract Background Isolated Prevotella intermedia, a rare gram-negative, rod-shaped, anaerobic bacterium, is rarely detected in clinical practice. It has been associated with infections of the oral cavity and female genital tract, but has never been detected in cerebrospinal fluid (CSF) of patients in China. Accurate detection of causative pathogens is still an arduous task owing to the difficult conditions of anaerobic bacterial culture. Isolated Prevotella intermedia can be detected by metagenomic next generation sequencing (mNGS) of the CSF. Correct diagnosis and antibiotic treatment can help patients avoid life-threatening events. Case presentation Herein, we describe the case of a 64-year-old Chinese woman who presented with typical features of meningoencephalitis. Routine CSF culture failed to identify the causative pathogen. Isolated Prevotella intermedia was detected by mNGS, and the patient was treated with antibacterial agents including ceftriaxone, vancomycin, moxifloxacin, meropenem, metronidazole, and linezolid. The patient underwent surgical treatment for abscess of left frontal parietal lobe, which was observed on magnetic resonance imaging (MRI) and was suspected to be caused by Prevotella intermedia. It was further confirmed that it was a secondary infection from the oral cavity, and the possible etiology might have been dental surgery. Treatment was rendered to the patient based on metagenomic test result, and her condition improved after two months. Conclusions This case highlights the role of mNGS in accurate diagnosis of patients with central nervous system infection. In particular, mNGS can be used to identify rare pathogens and confirm the diagnosis in patients with unknown etiology.

ABSTRACTAims Hemophore-like proteins sequester heme from host hemoproteins. We aimed to determine whether the host immune system can recognize not only Porphyromonas gingivalis HmuY but also its homologs expressed by other periodontopathogens, and how periodontitis influences the production of respective antibodies.Methods The reactivity of total bacterial antigens and purified proteins with serum IgG antibodies of 18 individuals with periodontitis and 17 individuals without periodontitis was examined by enzyme-linked immunosorbent assay (ELISA). To compare IgG reactivity between groups with and without periodontitis and within the various dilutions of sera, statistical analysis was performed using the Mann-Whitney U-test and two-way ANOVA test with the post-hoc Bonferroni test.Results Individuals with periodontitis produced IgG antibodies reacting more strongly not only with total P. gingivalis antigens (P = 0.0002; 1:400) and P. gingivalis HmuY (P = 0.0016; 1:100) but also with Prevotella intermedia PinA (P = 0.0059; 1:100), and with low efficiency with P. intermedia PinO (P = 0.0021; 1:100). No increase in the reactivity of IgG antibodies with Tannerella forsythia Tfo and P. gingivalis HusA was found in individuals with periodontitis.Conclusions Although hemophore-like proteins are structurally related, they are differentially recognized by the host immune system. Our findings point to specific antigens, mainly P. gingivalis HmuY and P. intermedia PinA, whose immunoreactivity could be further investigated to develop markers of periodontitis.

Aim: To evaluate the antibacterial activity of a-mangostin compounds against Prevotella intermedia bacteria using the diffusion method. Material and Methods: This type of research is a prospective analysis and in vivo experimental laboratory study. Research on a-mangostin as an antibacterial to inhibit the growth of the P. intermedia bacteria type (ATCC25611) with catalog number R3602016 was produced by Thermo Scientific, Inc. US. Fifty P. intermedia culture samples were chosen at random technique. The diffusion method is then used to assess the antibacterial activity of the a-mangostin compounds. Statistical Analysis: The data obtained were then examined using statistical assessment using the Kruskall-Wallis tests. Results: The results showed a concentration of a-mangostin which produces a zone of inhibition of the growth of P. intermedia with a diameter approaching the positive control group is the concentration of a-mangostin 100% then 50%, 25%, and 12.5%. Meanwhile, the concentration of a-mangostin which resulted in a zone of inhibition of the growth of P. intermedia bacteria with the same diameter as the negative control group was a-mangostin concentrations of 6.25%, 3.125%, 1.56%, and 0.78%. Conclusion: The a-mangostin compound has effective activity against the growth of the P. intermedia bacteria. [ABSTRACT FROM AUTHOR]

Background: Isolated Prevotella intermedia, a rare gram-negative, rod-shaped, anaerobic bacterium, is rarely detected in clinical practice. It has been associated with infections of the oral cavity and female genital tract, but has never been detected in cerebrospinal fluid (CSF) of patients in China. Accurate detection of causative pathogens is still an arduous task owing to the difficult conditions of anaerobic bacterial culture. Isolated Prevotella intermedia can be detected by metagenomic next generation sequencing (mNGS) of the CSF. Correct diagnosis and antibiotic treatment can help patients avoid life-threatening events. Case presentation: Herein, we describe the case of a 64-year-old Chinese woman who presented with typical features of meningoencephalitis. Routine CSF culture failed to identify the causative pathogen. Isolated Prevotella intermedia was detected by mNGS, and the patient was treated with antibacterial agents including ceftriaxone, vancomycin, moxifloxacin, meropenem, metronidazole, and linezolid. The patient underwent surgical treatment for abscess of left frontal parietal lobe, which was observed on magnetic resonance imaging (MRI) and was suspected to be caused by Prevotella intermedia. It was further confirmed that it was a secondary infection from the oral cavity, and the possible etiology might have been dental surgery. Treatment was rendered to the patient based on metagenomic test result, and her condition improved after two months. Conclusions: This case highlights the role of mNGS in accurate diagnosis of patients with central nervous system infection. In particular, mNGS can be used to identify rare pathogens and confirm the diagnosis in patients with unknown etiology. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the antibacterial and antibiofilm activities of the essential oils of Cuminum cyminum (CEO) and Foeniculum vulgare (FEO) on the clinically isolated Porphyromonas gingivalis (P. gingivalis) and Prevotella intermedia (P. intermedia). Subgingival plaque samples were collected from periodontal pockets (≥6 mm) of patients with generalized severe periodontitis. Microbiological and molecular tests were performed to isolate and confirm P. intermedia. A previously isolated P. gingivalis was used. The hydrodistillation method was used to extract the CEO and FEO. The antibacterial activity of the examined oils against the confirmed clinical strains was determined using the disc diffusion and broth dilution methods to determine the minimum inhibitory (MIC) and bactericidal concentrations (MBC). The antibiofilm properties of CEO and FEO were determined using the tube method. The t-test was used for comparisons of inhibition zones. Both CEO and FEO presented inhibition zones against both clinically isolated bacteria. The CEO and FEO revealed antibacterial activity with MIC values of 3.125 µL/mL and MBC of 6.25 µL/mL against P. gingivalis, respectively. In addition, their MIC and MBC against P. intermedia were 0.195 µL/mL and 3.125 µL/mL, respectively. Finally, the essential oils showed moderate to strong antibiofilm activities against the clinically isolated P. gingivalis and P. intermedia, respectively. This study supports the antibacterial and antibiofilm properties of CEO and FEO against clinically isolated P. gingivalis and P. intermedia. Further studies should focus on using these essential oils as an adjunct to periodontal therapy. [ABSTRACT FROM AUTHOR]

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Background: People globally are turning to herbal products to reconnect with nature. Cost efficacy and minimal side effects are the reasons for this changeover. This study assessed the effect of Amorphophallus paeoniifolius as an antimicrobial agent against Porphyromonas gingivalis, Prevotella intermedia, and Fusobacterium nucleatum. Aim: To determine and compare the antimicrobial activity of aqueous and ethanolic extracts of A. paeoniifolius on periodontal pathogens. Materials and Methods: Aqueous and ethanolic extracts of A. paeoniifolius were tested against the standard strains of the selected bacteria. Minimum inhibitory concentrations (MIC) and minimum bactericidal concentration (MBC) were used. These tests assessed the lowest concentrations of test agent, either by showing a lack of turbidity or by no or few bacterial growth colonies, respectively. In this study, tetracycline hydrochloride was used as the control group. Results: Aqueous and ethanolic extracts of A. paeoniifolius showed antibacterial activity at various concentrations against the selected organisms. While assessing the MBC, the aqueous and ethanolic extracts of A. paeoniifolius and tetracycline hydrochloride exhibited bactericidal activity against F. nucleatum at all concentrations. The ethanolic extract of Amorphophallus paeoniifolius and tetracycline hydrochloride showed bactericidal action, whereas the aqueous extract exhibited bacteriostatic action against P. gingivalis. The aqueous and ethanolic extracts of A. paeoniifolius showed bacteriostatic action, whereas tetracycline hydrochloride showed bactericidal action against P. intermedia. Conclusion: Both aqueous and ethanolic extracts of A. paeoniifolius showed antibacterial activity against standard strains of P. gingivalis, P. intermedia, and F. nucleatum. The ethanolic extract showed a significant antibacterial effect against the selected microorganisms when compared to the aqueous extract of A. paeoniifolius.

Background: Prevotella is a Gram-negative anaerobic bacilli. The phenotypic characteristics of the various species of Prevotella are similar, which often makes it difficult in routine differentiation and identification of all the species. Aim: The purpose of the study was to detect and compare presence of Prevotella intermedia, Prevotella nigrescens, Prevotella melaninogenica, and Prevotella loescheii in subgingival plaque samples of chronic periodontitis and healthy individuals. Materials and Methods: Two hundred and thirty-six subjects were considered consisting of chronic periodontitis (128) and healthy (108) individuals. Subgingival plaque sample was collected in reduced transport fluid and analyzed. DNA extraction and polymerase chain reaction (PCR) were performed for genus Prevotella followed by positive samples were considered for the detection of selected species through multiplex PCR using specific primers. Results: Out of 236 samples, 94.1% were positive for genus Prevotella. Out of 222 cases P. nigrescens showed the highest number of cases positive (59.5%) followed by P. melaninogenica (57.2%), P. intermedia (55.4%), and P. loescheii (40.1%). Species were analyzed individually between chronic periodontitis and healthy, P. intermedia, P. nigrescens, and P. loescheii showed greater positivity in healthy compared to chronic periodontitis. Positivity for P. melaninogenica was high in chronic periodontitis compared to healthy. Conclusion: The number of positive cases for species, when correlated with clinical parameters showed an increase in mean score for all clinical parameters assessed, suggesting the presence of variation in the prevalence of Prevotella species and geographic variation do exist in oral microflora. Findings suggest that they can be normal commensals and opportunistic.

Abstract Background Owing to the heterogeneity of microbiota among individuals and populations, only Fusobacterium nucleatum and Bacteroides fragilis have been reported to be enriched in colorectal cancer (CRC) in multiple studies. Thus, the discovery of additional bacteria contributing to CRC development in various populations can be expected. We aimed to identify bacteria associated with the progression of colorectal adenoma to carcinoma and determine the contribution of these bacteria to malignant transformation in patients of Han Chinese origin. Methods Microbiota composition was determined through 16S rRNA V3–V4 amplicon sequencing of autologous adenocarcinomas, adenomatous polyps, and non-neoplastic colon tissue samples (referred to as “tri-part samples”) in patients with CRC. Enriched taxa in adenocarcinoma tissues were identified through pairwise comparison. The abundance of candidate bacteria was quantified through genomic quantitative polymerase chain reaction (qPCR) in tissue samples from 116 patients. Associations of candidate bacteria with clinicopathological features and genomic and genetic alterations were evaluated through odds ratio tests. Additionally, the effects of candidate bacteria on CRC cell proliferation, migration, and invasion were evaluated through the co-culture of CRC cells with bacterial cells or with conditioned media from bacteria. Results Prevotella intermedia was overrepresented in adenocarcinomas compared with paired adenomatous polyps. Furthermore, co-abundance of P. intermedia and F. nucleatum was observed in tumor tissues. More notably, the coexistence of these two bacteria in adenocarcinomas was associated with lymph node involvement and distant metastasis. These two bacteria also exerted additive effects on the enhancement of the migration and invasion abilities of CRC cells. Finally, conditioned media from P. intermedia promoted the migration and invasion of CRC cells. Conclusion This report is the first to demonstrate that P. intermedia is enriched in colorectal adenocarcinoma tissues and enhances the migration and invasion abilities of CRC cells. Moreover, P. intermedia and F. nucleatum exert additive effects on the malignant transformation of colorectal adenomas into carcinomas. These findings can be used to identify patients at a high risk of malignant transformation of colorectal adenomas or metastasis of CRC, and they can accordingly be provided optimal clinical management.

Background: Prevotella is a Gram-negative anaerobic bacilli. The phenotypic characteristics of the various species of Prevotella are similar, which often makes it difficult in routine differentiation and identification of all the species. Aim: The purpose of the study was to detect and compare presence of Prevotella Intermedia, Prevotella nigrescens, Prevotella melaninogenica, and Prevotella loescheii in subgingival plaque samples of chronic periodontitis and healthy individuals. Materials and Methods: Two hundred and thirty-six subjects were considered consisting of chronic periodontitis (128) and healthy (108) individuals. Subgingival plaque sample was collected in reduced transport fluid and analyzed. DNA extraction and polymerase chain reaction (PCR) were performed for genus Prevotella followed by positive samples were considered for the detection of selected species through multiplex PCR using specific primers. Results: Out of 236 samples, 94.1% were positive for genus Prevotella. Out of 222 cases P. nigrescens showed the highest number of cases positive (59.5%) followed by P. melaninogenica (57.2%), P. intermedia (55.4%), and P. loescheii(40.1%). Species were analyzed individually between chronic periodontitis and healthy, P. intermedia, P. nigrescens, and P. loescheii showed greater positivity in healthy compared to chronic periodontitis. Positivity for P. melaninogenica was high in chronic periodontitis compared to healthy. Conclusion: The number of positive cases for species, when correlated with clinical parameters showed an increase in mean score for all clinical parameters assessed, suggesting the presence of variation in the prevalence of Prevotella species and geographic variation do exist in oral microflora. Findings suggest that they can be normal commensals and opportunistic. [ABSTRACT FROM AUTHOR]

Objective: To analyse the pan-genome of three black-pigmented periodontal pathogens: Porphyromonas gingivalis, Prevotella intermedia and Prevotella nigrescens. Methods: Pan-genome analyses of 66, 33 and 5 publicly available whole-genome sequences of P. gingivalis, P. intermedia and P. nigrescens, respectively, were performed using Pan-genome Analysis Pipeline software (version 1.2.1; Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, PR China). Phylogenetic trees were constructed based on the entire pangenome and single nucleotide polymorphisms within the core genome. The distribution and abundance of virulence genes in the core and dispensable genomes were also compared in the three species. Results: All three species possess an open pan-genome. The core genome of P. gingivalis, P. inter)media and P. nigrescens included 1001, 1514 and 1745 orthologous groups, respectively, which were mainly related to basic cellular functions such as metabolism. The dispensable genome of P. gingivalis, P. intermedia and P. nigrescens was composed of 2814, 2689 and 906 orthologous groups, respectively, and it was enriched in genes involved in pathogenicity or with unknown functions. Phylogenetic trees presented a clear separation of P. gingivalis, P. intermedia and P. nigrescens, verifying the reclassification of the black-pigmented species. Furthermore, the three species shared almost the same virulence factors involved in adhesion, proteolysis and evasion of host defences. Some of these virulence genes were conserved across species whereas others belonged to the dispensable genome, which might be acquired through horizontal gene transfer. Conclusion: This study highlighted the usefulness of pan-genome analysis to infer evolutionary cues for black-pigmented species, indicating their homology and phylogenomic di [ABSTRACT FROM AUTHOR]

ABSTRACT Bacteria have to persist under low iron conditions in order to adapt to the nutritional immunity of a host. Since the knowledge of iron stimulon of Bacteroidetes is sparse, we examined oral (Porphyromonas gingivalis and Prevotella intermedia) and gut (Bacteroides thataiotaomicron) representatives for their ability to adapt to iron deplete and iron replete conditions. Our transcriptomics and comparative genomics analysis show that many iron-regulated mechanisms are conserved within the phylum. They include genes upregulated in low iron, as follows: fldA (flavodoxin), hmu (hemin uptake operon), and loci encoding ABC transporters. Downregulated genes were frd (ferredoxin), rbr (rubrerythrin), sdh (succinate dehydrogenase/fumarate reductase), vor (oxoglutarate oxidoreductase/dehydrogenase), and pfor (pyruvate:ferredoxin/flavodoxin oxidoreductase). Some genus-specific mechanisms, such as the sus of B. thetaiotaomicron coding for carbohydrate metabolism and the xusABC coding for xenosiderophore utilization were also identified. While all bacteria tested in our study had the nrfAH operon coding for nitrite reduction and were able to reduce nitrite levels present in culture media, the expression of the operon was iron dependent only in B. thetaiotaomicron. It is noteworthy that we identified a significant overlap between regulated genes found in our study and the B. thetaiotaomicron colitis study (W. Zhu, M. G. Winter, L. Spiga, E. R. Hughes et al., Cell Host Microbe 27:376–388, 2020, http://dx.doi.org/10.1016/j.chom.2020.01.010). Many of those commonly regulated genes were also iron regulated in the oral bacterial genera. Overall, this work points to iron being the master regulator enabling bacterial persistence in the host and paves the way for a more generalized investigation of the molecular mechanisms of iron homeostasis in Bacteroidetes. IMPORTANCE Bacteroidetes are an important group of anaerobic bacteria abundant both in the oral and gut microbiomes. Although iron is a required nutrient for most living organisms, the molecular mechanisms of adaptation to the changing levels of iron are not well known in this group of bacteria. We defined the iron stimulon of Bacteroidetes by examination of the transcriptomic response of Porphyromonas gingivalis and Prevotella intermedia (both belong to the oral microbiome) and Bacteroidetes thetaiotaomicron (belongs to the gut microbiome). Our results indicate that many of the iron-regulated operons are shared among the three genera. Furthermore, using bioinformatics analysis, we identified a significant overlap between our in vitro studies and transcriptomic data derived from a colitis study, thus underscoring the biological significance of our work. Defining the iron-dependent stimulon of Bacteroidetes can help to identify the molecular mechanisms of iron-dependent regulation as well as better understand the persistence of the anaerobes in the human host.

Aim: The aim of the present study was to detect and correlate the levels of Prevotella Intermedia with clinical parameters after nonsurgical periodontal therapy (NSPT) in chronic periodontitis female patients with or without oral contraceptive pills, using quantitative polymerase chain reaction (Q-PCR) method. Materials and Methods: Sixty female patients equally divided into three groups, i.e. Periodontally health female patients (Group -I), females with Chronic Periodontitis (CP) without taking oral contraceptive pills(OCP) (Group-II), females with Chronic Periodontitis (CP) taking oral contraceptive pills(OCP) (Group-III) and were correlated for the presence of P. Intermedia in the respective groups. PPD, CAL, and saliva samples for microbiological evaluation were assessed at baseline, 1-, and 3-month post-NSPT. Results: Significant reduction of PPD was found 2.4 ± 0.48mm versus 1.4 ± 0.37 mm in Group I, 4.8 ± 0.74 versus 2.8 ± 0.53 mm in Group II, and 6.6 ± 0.48 mm versus 4.4 ± 0.39 mm in Group III post-NSPT at 3 months. Similarly, a notable reduction of CAL was exhibited in both Group II (5.4 ± 0.65 vs. 3.1 ± 0.42 mm) and Group III (6.6 ± 0.48 vs 3.6 ± 0.35 mm) patients after NSPT at 3 months. A greater reduction of P. Intermedia concentrations was observed in both Group II and Group III at 3-month post-NSPT. Conclusion: The substantial improvement of clinical parameters was found to be in correlation with the load of P. Intermedia, which was reduced more in Group II than in Group III, emphasizing the applicability and sensitivity of Q-PCR method for its assessment. [ABSTRACT FROM AUTHOR]

Aims: Hemophore-like proteins sequester heme from host hemoproteins. We aimed to determine whether the host immune system can recognize not only Porphyromonas gingivalis HmuY but also its homologs expressed by other periodontopathogens, and how periodontitis influences the production of respective antibodies. Methods: The reactivity of total bacterial antigens and purified proteins with serum IgG antibodies of 18 individuals with periodontitis and 17 individuals without periodontitis was examined by enzyme-linked immunosorbent assay (ELISA). To compare IgG reactivity between groups with and without periodontitis and within the various dilutions of sera, statistical analysis was performed using the Mann-Whitney U-test and two-way ANOVA test with the post-hoc Bonferroni test. Results: Individuals with periodontitis produced IgG antibodies reacting more strongly not only with total P. gingivalis antigens (P = 0.0002; 1:400) and P. gingivalis HmuY (P = 0.0016; 1:100) but also with Prevotella intermedia PinA (P = 0.0059; 1:100), and with low efficiency with P. intermedia PinO (P = 0.0021; 1:100). No increase in the reactivity of IgG antibodies with Tannerella forsythia Tfo and P. gingivalis HusA was found in individuals with periodontitis. Conclusions: Although hemophore-like proteins are structurally related, they are differentially recognized by the host immune system. Our findings point to specific antigens, mainly P. gingivalis HmuY and P. intermedia PinA, whose immunoreactivity could be further investigated to develop markers of periodontitis. [ABSTRACT FROM AUTHOR]

Background: People globally are turning to herbal products to reconnect with nature. Cost efficacy and minimal side effects are the reasons for this changeover. This study assessed the effect of Amorphophallus paeoniifolius as an antimicrobial agent against Porphyromonas gingivalis, Prevotella intermedia, and Fusobacterium nucleatum. Aim: To determine and compare the antimicrobial activity of aqueous and ethanolic extracts of A. paeoniifolius on periodontal pathogens. Materials and Methods: Aqueous and ethanolic extracts of A. paeoniifolius were tested against the standard strains of the selected bacteria. Minimum inhibitory concentrations (MIC) and minimum bactericidal concentration (MBC) were used. These tests assessed the lowest concentrations of test agent, either by showing a lack of turbidity or by no or few bacterial growth colonies, respectively. In this study, tetracycline hydrochloride was used as the control group. Results: Aqueous and ethanolic extracts of A. paeoniifolius showed antibacterial activity at various concentrations against the selected organisms. While assessing the MBC, the aqueous and ethanolic extracts of A. paeoniifolius and tetracycline hydrochloride exhibited bactericidal activity against F. nucleatum at all concentrations. The ethanolic extract of Amorphophallus paeoniifolius and tetracycline hydrochloride showed bactericidal action, whereas the aqueous extract exhibited bacteriostatic action against P. gingivalis. The aqueous and ethanolic extracts of A. paeoniifolius showed bacteriostatic action, whereas tetracycline hydrochloride showed bactericidal action against P. intermedia. Conclusion: Both aqueous and ethanolic extracts of A. paeoniifolius showed antibacterial activity against standard strains of P. gingivalis, P. intermedia, and F. nucleatum. The ethanolic extract showed a significant antibacterial effect against the selected microorganisms when compared to the aqueous extract of A. paeoniifolius. [ABSTRACT FROM AUTHOR]

Background: As periodontitis progresses, the oral microbiota community changes dynamically. In this study, we evaluated the dominant bacteria and their roles in the potential pathway in young males with stage III periodontitis. Methods: 16S rRNA sequencing was performed to evaluate variations in the composition of oral bacteria between males with stage I and III periodontitis and identify the dominant bacteria of each group. Function prediction was obtained based on 16S rRNA sequencing data. The inhibitor of the predominant pathway for stage III periodontitis was used to investigate the role of the dominant bacteria in periodontitis in vivo and in vitro. Results: Chao1 index, Observed Species and Phylogenetic Diversity (PD) whole tree values were significantly higher in the stage III periodontitis group. β-diversity suggested that samples could be divided according to the stages of periodontitis. The dominant bacteria in stage III periodontitis were Prevotella, Prevotella_7, and Dialister, whereas that in stage I periodontitis was Cardiobacterium. KEGG analysis predicted that variations in the oral microbiome may be related to the NOD-like receptor signaling pathway. The inhibitor of this pathway, NOD-IN-1, decreased P. intermedia -induced Tnf-α mRNA expression and increased P. intermedia -induced Il-6 mRNA expression, consistent with the ELISA results. Immunohistochemistry confirmed the downregulation of TNF-α and IL-6 expressions by NOD-IN-1 in P. intermedia-induced periodontitis. Conclusion: The composition of the oral bacteria in young males varied according to the stage of periodontitis. The species richness of oral microtia was greater in young males with stage III periodontitis than those with stage I periodontitis. Prevotella was the dominant bacteria in young males with stage III periodontitis, and inhibition of the NOD-like receptor signaling pathway can decrease the periodontal inflammation induced by P. intermedia. [ABSTRACT FROM AUTHOR]

Objective To study the distribution of pathogenic infection and relevance in combined periodontal-endodontic lesions of periodontal origin,and provide the basis for clinical treatment. Methods Totally 43 cases of combined periodontal-endodontic lesions of periodontal origin from Jan. 2018 to Jun. 2020 treated in the hospital were selected, including a total of 43 teeth as the experimental group. Another 41 teeth from 41 cases with severe periodontitis during the same period were set as the control group. subgingival plaque samples and root canal samples of ill teeth were made for test. Quantitative Real-time PCR was used to detect the quantity of Tannerella forsythia (Tf), Porphyromonas gingivalis (Pg), Fusobacterium nucleatum (Fn), Prevotella intermedia (Pi), Treponema denticola (Td), Digestive streptococcus (Ds), Enterococcus faecalis (Ef), Porphyromanus endodontics (Pe). Results There was no significant difference in the quantity of Digestive streptococcus and Porphyromanus endodontics in the root canal tissue and subgingival plaque of the experimental group (Ρ>0.05), other six pathogenic bacteria in the subgingival plaque samples was significantly higher than that from the root canal tissue (P

Background/purpose: Bacterial biofilms formed on the surface of tissues and biomaterials are major causes of chronic infections in humans. Among them, Porphyromonas gingivalis (P. gingivalis) and Prevotella intermedia (P. intermedia) are anaerobic pathogens causing dental infections associated with periodontitis. In this study, we evaluated the killing effect and underlying mechanisms of direct current (DC) as an antimicrobial method in vitro. Materials and methods: We chose P. gingivalis and P. intermedia in different states to make comparisons of the killing effect of DC. By viable bacteria counting, fluorescent live/dead staining, reactive oxygen species (ROS) assay, addition of ROS scavenger DMTU and mRNA expression assay of ROS scavenging genes, the role of ROS in the killing effect was explored. Results: The planktonic and biofilm states of two bacteria could be effectively killed by low-intensity DC. For the killing effect of 1000 μA DC, there were significant differences whether on planktonic P. gingivalis and P. intermedia (mean killing values: 2.40 vs 2.62 log10 CFU/mL) or on biofilm state of those (mean killing values: 0.63 vs 0.98 log10 CFU/mL). 1000 μA DC greatly induced ROS production and the mRNA expression of ROS scavenging genes. DMTU could partially decrease the killing values of DC and downregulate corresponding gene’s expression. Conclusion: 1000 μA DC can kill P. gingivalis and P. intermedia in two states by promoting overproduction of ROS, and P. intermedia is more sensitive to DC than P. gingivalis. These findings indicate low-intensity DC may be a promising approach in treating periodontal infections.

Background: The development and progression of periodontal diseases is a result of the dynamic interaction of microorganisms within their habitat, and changes in this habitat generate a dysbiotic state. Fusobacterium nucleatum and Prevotella intermedia are bridging microorganisms between the pioneer communities and other microorganisms responsible for periodontitis such as Porphyromonas gingivalis. Tetracycline hydrochloride (TTC-HCl) is commonly used as a coadjutant in periodontal treatment in the form of an antiseptic. However, there are no clear dilution or concentration protocols. Objective: This study aimed to evaluate the in vitro antimicrobial activity of TTC-HCl diluted in sterile water, saline solution, and 2% lidocaine with epinephrine 1:80,000 at concentration of 125, 250, and 500 mg, at three time points– 30, 60, and 120 s – on P. intermedia, F. nucleatum, and P. gingivalis using the Kelsey–Maurer technique. Materials and Methods: The antimicrobial activity of TTC-HCl was evaluated at the proposed concentrations and times, dissolved in the different vehicles at pH 1.9 and 7.0, on F. nucleatum, P. intermedia, and P. gingivalis. The Kelsey–Maurer test was used to verify the presence or absence of colony-forming units. Each test was performed in triplicates with its respective viability controls. Results: Inhibition of F. nucleatum, P. intermedia, and P. gingivalis was achieved with TTC-HCl at all concentrations, dissolved in distilled water, saline solution, and 2% lidocaine with epinephrine 1:80,000 for all times. Conclusions: The results show that TTC-HCl is a good antimicrobial alternative against F. nucleatum, P. intermedia, and P. gingivalis regardless of the vehicle in which it was dissolved, concentration, pH, or time used in this investigation.

ObjectivePeriodontitis progression is related to the dynamic dysbiosis of oral microbiome. We identified the dominant bacteria and the potential pathway in young women with stage-III periodontitis.Materials and methodsSamples of subgingival plaque were collected from 26 young women with periodontitis (20 with stage-I and 6 with stage-III). Using 16S rRNA-sequencing, we determined the variation in oral bacterial communities of the two groups, and identified the dominant bacteria of each group. We used the Kyoto Encyclopedia of Genes and Genomes (KEGG) database to evaluate the signaling pathways related to the difference in oral bacterial composition. The role of the dominant bacteria of stage-III periodontitis was investigated in vivo and in vitro using an endoplasmic reticulum stress inhibitor.ResultsYoung women with stage-I periodontitis had higher values for the Chao1 Index, Observed Species and Phylogenetic Diversity Whole Tree Index than those for women with stage-III periodontitis. β-diversity analyses revealed that samples could be divided into different groups according to the periodontitis stage. The most representative biomarkers of stage-III periodontitis in young women were bacteria of the phylum Bacteroidetes, its order, family and genera Bacteroidales, Prevotellaceae and Prevotella. The KEGG database revealed that the change in oral bacterial composition of young women with stage-III periodontitis may be related to protein processing in an endoplasmic reticulum pathway. Salubrinal (an endoplasmic reticulum stress regulator) controlled expression of Runx2, Col1a1, Ocn in mouse bone-marrow mesenchymal cells. Salubrinal administration showed that moderate endoplasmic reticulum stress inhibited alveolar bone loss in periodontitis induced by Prevotella intermedia lipopolysaccharide.ConclusionDifferences between periodontitis stages were noted and bacteria of Prevotella species were abundant in young women with stage-III periodontitis. This phenomenon was related to protein processing in an endoplasmic reticulum pathway.

Purpose: To compare the qualitative and quantitative profile of Epstein-Barr virus (EBV) at external and internal implant surfaces between participants with peri-implantitis and healthy peri-implant tissues and to quantitatively assess the relation between EBV and periopathogens inside the microbiologic profile associated with peri-implantitis. Materials and Methods: Microbiologic specimens were retrieved from 84 patients wearing 190 implants to estimate the levels of EBV and 10 periopathogens in the peri-implant pocket and internal-implant connection using quantitative polymerase chain reaction. Results: The study sample consisted of 113 healthy and 77 peri-implantitis--affected implants. Statistical significance was not reached in EBV prevalence between peri-implantitis and healthy controls. EBV-positive participants demonstrated higher levels of Prevotella intermedia (Pi) and Campylobacter rectus (Cr) compared with EBV-negative participants. A positive correlation was demonstrated among EBV and Tannerella forsythia (Tf), Parvimonas micra (Pm), Fusobacterium nucleatum (Fn), and Cr levels in peri-implantitis--affected implants, while healthy controls demonstrated a positive correlation between EBV and Aggregatibacter actinomycetemcomitans (Aa), Pi, and Pm. Conclusion: EBV cannot be considered as a microbiologic marker of peri-implantitis. However, EBV could be considered as a risk factor and a peri-implantitis enhancer based on its positive correlations with pathogens associated with peri-implantitis. [ABSTRACT FROM AUTHOR]

Background: Owing to the heterogeneity of microbiota among individuals and populations, only Fusobacterium nucleatum and Bacteroides fragilis have been reported to be enriched in colorectal cancer (CRC) in multiple studies. Thus, the discovery of additional bacteria contributing to CRC development in various populations can be expected. We aimed to identify bacteria associated with the progression of colorectal adenoma to carcinoma and determine the contribution of these bacteria to malignant transformation in patients of Han Chinese origin. Methods: Microbiota composition was determined through 16S rRNA V3–V4 amplicon sequencing of autologous adenocarcinomas, adenomatous polyps, and non-neoplastic colon tissue samples (referred to as "tri-part samples") in patients with CRC. Enriched taxa in adenocarcinoma tissues were identified through pairwise comparison. The abundance of candidate bacteria was quantified through genomic quantitative polymerase chain reaction (qPCR) in tissue samples from 116 patients. Associations of candidate bacteria with clinicopathological features and genomic and genetic alterations were evaluated through odds ratio tests. Additionally, the effects of candidate bacteria on CRC cell proliferation, migration, and invasion were evaluated through the co-culture of CRC cells with bacterial cells or with conditioned media from bacteria. Results: Prevotella intermedia was overrepresented in adenocarcinomas compared with paired adenomatous polyps. Furthermore, co-abundance of P. intermedia and F. nucleatum was observed in tumor tissues. More notably, the coexistence of these two bacteria in adenocarcinomas was associated with lymph node involvement and distant metastasis. These two bacteria also exerted additive effects on the enhancement of the migration and invasion abilities of CRC cells. Finally, conditioned media from P. intermedia promoted the migration and invasion of CRC cells. Conclusion: This report is the first to demonstrate that P. intermedia is enriched in colorectal adenocarcinoma tissues and enhances the migration and invasion abilities of CRC cells. Moreover, P. intermedia and F. nucleatum exert additive effects on the malignant transformation of colorectal adenomas into carcinomas. These findings can be used to identify patients at a high risk of malignant transformation of colorectal adenomas or metastasis of CRC, and they can accordingly be provided optimal clinical management. [ABSTRACT FROM AUTHOR]

The hypothesis of the present study was that nitro-fatty acids (NO 2 -FAs) would suppress inflammation associated with periodontal disease. To test this hypothesis, we investigated the influence of nitrooleic acid, a prototypical NO 2 -FA, on the inflammatory response of murine macrophages activated with lipopolysaccharide (LPS) from Prevotella intermedia , a pathogen associated the etiology of different types of periodontal diseases. LPS was prepared from P. intermedia cells by using phenol-water protocol. Culture supernatants were assayed for nitric oxide (NO), interleukin-1β (IL-1β), and IL-6. Real-time polymerase chain reaction and immunoblotting analyses were performed to quantify messenger RNA and protein expression, respectively. The secreted embryonic alkaline phosphatase reporter assay was performed to measure NF-κB activation. The transcription factor assay kit was used to measure DNA-binding of NF-κB subunits. Findings obtained from the present study revealed that nitrooleic acid suppresses the generation and messenger RNA expression of inducible NO synthase-derived NO, IL-1β, and IL-6 in RAW264.7 cells activated with P. intermedia LPS and promotes macrophage polarization toward anti-inflammatory M2 phenotype. We also found that nitrooleic acid exerts its effect via heme oxygenase-1 induction and suppression of NF-κB signaling. The inhibition of NO and proinflammatory cytokine production by nitrooleic acid was independent from PPAR-γ, JNK, p38, and STAT1/3. Nitrooleic acid may represent a novel class of agent as a host modulator which has therapeutic benefit in periodontal disease, though more work is needed to confirm this. Proposed cellular mechanisms involved in the inhibition of Prevotella intermedia lipopolysaccharide (LPS)-induced production of nitric oxide (NO), interleukin-1β (IL-1β), and IL-6 by nitrooleic acid (OA-NO 2) in RAW264.7 cells. The generation and messenger RNA expression of inducible NO synthase-derived NO, IL-1β, and IL-6 induced by P. intermedia LPS were suppressed by OA-NO 2. OA-NO 2 encouraged the switch from proinflammatory M1 macrophages to anti-inflammatory M2 macrophage phenotype in cells activated with LPS. OA-NO 2 exerts its effect via heme oxygenase-1 induction and suppression of NF-κB signaling. [Display omitted] [ABSTRACT FROM AUTHOR]

Background: The study focuses on evaluating the parasitic potential of novel metronidazole analogs using computational methods. Specifically, it aims to target key enzymes of oral anaerobes, including UDP-N-acetylglucosamine 1-carboxyvinyltransferase (MurA) of Fusobacterium nucleatum and DNA topoisomerase (Topo) of Prevotella intermedia., Objective: The objective is to assess the pharmacokinetic and toxicity properties of 368 novel nitroimidazole candidates through virtual screening. Additionally, the study aims to determine the binding affinity of the most promising candidates with the target proteins through molecular docking analyses., Methods: A combinatorial library of nitroimidazole candidates was constructed, and virtual screening was performed. Molecular docking analyses were conducted to evaluate the binding affinity of selected compounds with MurA and Topo. Further investigation involved molecular dynamic simulation to assess the stability of the compounds within the active sites of MurA and Topo., Results: All selected compounds exhibited activity against both MurA and Topo. Among them, Mnz11, Mnz12, and Mnz15 demonstrated the lowest binding free energies and IC50 values. Molecular dynamic simulation indicated that these three compounds remained stable within the active sites of MurA and Topo, with RMSD values consistently below 2Å. Additionally, the antibacterial potential of the most potent compound, Mnz15, was evaluated against a series of oral microbes., Conclusion: The study concludes that the newly identified nitroimidazole candidates show promise as anti-parasitic agents, based on their activity against key enzymes of oral anaerobes and their pharmacokinetic properties evaluated through computational methods., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

Objective: To determine the correlation between levels of methyl mercaptan (CH3SH) hydrogen sulfide (H2S), the proportion of Prevotella intermedia (Pi), and matrix metalloproteinase-8 (MMP-8) gene expression levels in periodontitis patients accompanied by halitosis. Material and Methods: Samples were obtained from gingival crevicular fluid (GCF) in the deepest pocket and by swabbing in the tongue coating area in patients with periodontitis presenting with halitosis (n = 23) and healthy subjects as controls (n = 7). The values of CH3SH and H2S were obtained using Oral Chroma. The proportion of Pi and MMP-8 expression levels were evaluated using PCR-RT. All the result was statistically analyzed using SPSS software. Results: The levels of CH3SH and H2S in participants with PD ≥ 6 mm showed a robust negative correlation with the proportion of P. intermedia in GCF and tongue coating. No statistically significant association was detected between CH3SH and H2S levels and MMP-8 expression levels (p>0.05). Conclusion: There is no association between CH3SH and H2S levels, the proportion of P. intermedia, and MMP-8 expression in patients with periodontitis accompanied by halitosis.

This study aimed to investigate the antibacterial and antibiofilm activities of the essential oils of Cuminum cyminum (CEO) and Foeniculum vulgare (FEO) on the clinically isolated Porphyromonas gingivalis (P. gingivalis) and Prevotella intermedia (P. intermedia). Subgingival plaque samples were collected from periodontal pockets (≥6 mm) of patients with generalized severe periodontitis. Microbiological and molecular tests were performed to isolate and confirm P. intermedia. A previously isolated P. gingivalis was used. The hydrodistillation method was used to extract the CEO and FEO. The antibacterial activity of the examined oils against the confirmed clinical strains was determined using the disc diffusion and broth dilution methods to determine the minimum inhibitory (MIC) and bactericidal concentrations (MBC). The antibiofilm properties of CEO and FEO were determined using the tube method. The t-test was used for comparisons of inhibition zones. Both CEO and FEO presented inhibition zones against both clinically isolated bacteria. The CEO and FEO revealed antibacterial activity with MIC values of 3.125 µL/mL and MBC of 6.25 µL/mL against P. gingivalis, respectively. In addition, their MIC and MBC against P. intermedia were 0.195 µL/mL and 3.125 µL/mL, respectively. Finally, the essential oils showed moderate to strong antibiofilm activities against the clinically isolated P. gingivalis and P. intermedia, respectively. This study supports the antibacterial and antibiofilm properties of CEO and FEO against clinically isolated P. gingivalis and P. intermedia. Further studies should focus on using these essential oils as an adjunct to periodontal therapy.

The Prevotella genus is a normal constituent of the oral microbiota, and is commonly isolated from mechanically treated polymicrobial infections. However, antibiotic treatment is necessary for some patients. This study compared the antibiotic susceptibility and the presence of resistance genes in clinical oral isolates of P. intermedia, P. nigrescens, and P. melaninogenica. Antibiotic susceptibility was assessed using the agar dilution method. PCR confirmed the species and resistance gene frequency in the Prevotella species. The frequencies of species P. intermedia, P. nigrescens, and P. melaninogenica were 30.2%, 45.7%, and 24.1%, respectively. No isolates of P. intermedia were resistant to amoxicillin/clavulanic acid, tetracycline, or clindamycin. P. nigrescens and P. melaninogenica were resistant to amoxicillin/clavulanic acid and tetracycline at frequencies of 40% and 20%, respectively. P. intermedia was resistant to metronidazole at a frequency of 30%, P. nigrescens at 20%, and P. melaninogenica at 40%. P. nigrescens and P. melaninogenica were resistant to 50% and 10% clindamycin, respectively. The gene most frequently detected was tetQ, at 43.3%, followed by tetM at 36.6%, blaTEM at 26.6%, ermF at 20%, cfxA, cfxA2, and nimAB at 16.6%, and nimAEFI at 3.3%. P. nigrescens was the species with the highest resistance to antibiotics such as amoxicillin/clavulanic acid, amoxicillin, and clindamycin, in addition to being the species with the largest number of genes compared to P. intermedia and P. melaninogenica. [ABSTRACT FROM AUTHOR]

Pendahuluan: Prevotella intermedia merupakan salah satu bakteri utama pada penyakit periimplantitis. Periimplantitis merupakan inflamasi jaringan lunak dan keras disekitar implan yang dapat dicegah menggunakan ekstrak tanaman antibakteri. Salah satunya yaitu kulit jeruk nipis, yang dapat menghambat proses inflamasi karena mengandung alkaloid, steroid, saponin, flavonoid, tanin sebagai senyawa antibakteri. Penelitian ini bertujuan untuk mengetahui peranan antibakteri kulit jeruk nipis dengan mengukur Konsentrasi Hambat Minimal (KHM) dan Konsentrasi Bunuh Minimal (KBM) ekstrak kulit jeruk nipis terhadap pertumbuhan bakteri Prevotella intermedia. Metode: Eksperimental laboratorium dengan rancangan penelitian post test only control group design. Pengujian KHM dan KBM dilakukan dengan metode dilusi, kulit jeruk nipis dimaserasi menggunakan pelarut etanol 70% sehingga didapatkan ekstrak kulit jeruk nipis dengan konsentrasi 0,78, 1,56, 3,125, 6,25, 12,5, 25, 50, dan 100% dengan chlorhexidine sebagai kontrol positif dan akuades sebagai kontrol negatif. Media kultur bakteri menggunakan Tripton Soya Agar. Hasil: Berdasarkan analisis statistik menggunakan Kruskal Wallis menunjukkan perbedaan penurunan jumlah koloni bakteri yang signifikan (p=0,0001) pada KBM dan KHM dari berbagai konsentrasi ekstrak kulit jeruk nipis terhadap pertumbuhan bakteri Prevotella intermedia, uji lanjutan Mann Whitney menunjukkan perbedaan penurunan jumlah koloni bakteri yang signifikan (p=0,021) antar masing-masing konsentrasi dan kelompok kontrol. Simpulan: Konsentrasi hambat minimal ekstrak kulit jeruk nipis terhadap bakteri Prevotella intermedia 12,5%, konsentrasi bunuh minimalnya 25%. Kata kunci: agen antibakteri; ekstrak jeruk nipis; Prevotella intermedia ABSTRACT Introduction: Prevotella intermedia is one of the main bacteria in periimplantitis. Periimplantitis is inflammation of the soft and hard tissues around the implant that can be prevented using antibacterial plant extracts. One of them is lime peel, which can inhibit the inflammatory process due to its alkaloids, steroids, saponins, flavonoids, and tannins as antibacterial compounds. This study was aimed to determine the antibacterial activity of lime peel by measuring the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of lime peel extract on the growth of Prevotella intermedia. Methods: Experimental laboratory with post-test only control group design. The MIC and MBC tests were performed by the dilution method. The lime peel was macerated using 70% ethanol solvent to obtain lime peel extract with concentrations of 0.78, 1.56, 3.125, 6.25, 12.5, 25, 50, and 100% with chlorhexidine as a positive control and aquadest as a negative control. Bacterial culture media using Tripton Soy Agar. Results: Based on statistical analysis using Kruskal Wallis showed a significant difference in the decrease of the number of bacterial colonies (p=0.0001) in MBC and MIC from various concentrations of lime peel extract on the growth of Prevotella intermedia bacteria. Furthermore, the Mann Whitney follow-up test showed differences in the decrease of the number of bacterial colonies, which was significant (p=0.021) between each concentration and control group. Conclusions: The minimum inhibitory concentration of lime peel extract towards the growth of Prevotella intermedia was 12.5%, with the minimum bactericidal concentration of 25%. Keywords: antibacterial agent; citrus extracts; Prevotella intermedia

For the treatment of periodontitis stage III/IV, a quadrant/week-wise debridement (Q-SRP) was compared with three full-mouth approaches: full-mouth scaling (FMS, accelerated Q-SRP within 24 h), full-mouth scaling with chlorhexidine-based disinfection (FMD), and FMD with adjuvant erythritol air polishing (FMDAP). The objective of this prospective, randomized study (a substudy of ClinicalTrials.gov, identifier: NCT03509233) was to compare the clinical and microbiological effects of the treatments. In total, 105 patients were randomized to one of the four aforementioned treatment groups, with n = 25, 28, 27, and 25 patients allocated to each group, respectively. At baseline and 3 and 6 months after treatment, the clinical parameters, including the pocket probing depths, clinical attachment level, and bleeding on probing, were recorded, and the prevalence of the total bacteria and four periodontal pathobionts (Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, and Tannerella forsythia) was determined using real-time quantitative PCR. Concerning the clinical outcomes, all the treatment modalities were effective, but the full-mouth approaches, especially FMDAP, were slightly superior to Q-SRP. Using the FMD approach, the reduction in the bacterial load and the number of pathobionts was significantly greater than for FMS, followed by Q-SRP. FMDAP was the least effective protocol for microbial reduction. However, after a temporary increase 3 months after therapy using FMDAP, a significant decrease in the key pathogen, P. gingivalis, was observed. These findings were not consistent with the clinical results from the FMDAP group. In conclusion, the dynamics of bacterial colonization do not necessarily correlate with clinical outcomes after full-mouth treatments for periodontitis stage III/IV. [ABSTRACT FROM AUTHOR]

Background: The development and progression of periodontal diseases is a result of the dynamic interaction of microorganisms within their habitat, and changes in this habitat generate a dysbiotic state. Fusobacterium nucleatum and Prevotella intermedia are bridging microorganisms between the pioneer communities and other microorganisms responsible for periodontitis such as Porphyromonas gingivalis. Tetracycline hydrochloride (TTC-HCl) is commonly used as a coadjutant in periodontal treatment in the form of an antiseptic. However, there are no clear dilution or concentration protocols. Objective: This study aimed to evaluate the in vitro antimicrobial activity of TTC-HCl diluted in sterile water, saline solution, and 2% lidocaine with epinephrine 1:80,000 at concentration of 125, 250, and 500 mg, at three time points– 30, 60, and 120 s – on P. intermedia, F. nucleatum, and P. gingivalis using the Kelsey–Maurer technique. Materials and Methods: The antimicrobial activity of TTC-HCl was evaluated at the proposed concentrations and times, dissolved in the different vehicles at pH 1.9 and 7.0, on F. nucleatum, P. intermedia, and P. gingivalis. The Kelsey–Maurer test was used to verify the presence or absence of colony-forming units. Each test was performed in triplicates with its respective viability controls. Results: Inhibition of F. nucleatum, P. intermedia, and P. gingivalis was achieved with TTC-HCl at all concentrations, dissolved in distilled water, saline solution, and 2% lidocaine with epinephrine 1:80,000 for all times. Conclusions: The results show that TTC-HCl is a good antimicrobial alternative against F. nucleatum, P. intermedia, and P. gingivalis regardless of the vehicle in which it was dissolved, concentration, pH, or time used in this investigation. [ABSTRACT FROM AUTHOR]

Background: Periodontitis, an inflammatory disease of multifactorial etiology, has bacteria playing an essential role in its pathogenesis. Prevotella intermedia plays an important role in disease initiation and progression. Objectives: The objective was to detect immunoglobulin G (IgG) antibodies against P. intermedia in blood of periodontally healthy individuals and patients with chronic periodontitis and compare their levels. Materials and Methodology: A total of 72 subjects were included, 36 subjects in the healthy group and 36 subjects in the chronic periodontitis group. Subgingival plaque sample and blood sample were obtained from each study subject. Samples were processed in the Department of Microbiology and Immunology. P. intermedia were confirmed using the culture method and serum IgG levels were assessed using ELISA (enzyme-linked immunosorbent assay) technique. Comparison between healthy and chronic periodontitis groups was done using independent t-test. Results: IgG levels against P. intermedia were more in the chronic periodontitis group compared to the healthy group, and the difference was statistically significant. Interpretations and Conclusions: Increased levels of IgG antibodies against P. intermedia are associated with periodontal disease. This elevated antibody activity might help to neutralize the effects of the bacterium. IgG antibody level against P. intermedia is a promising indicator in the serological diagnosis of periodontal disease. In chronic periodontitis, the antibody titer in the patient's serum against P. intermedia is raised and could be used as a diagnostic aid.

Aim: The study aimed at comparing the antimicrobial activity of mycelium and spore of Ganoderma lucidum on Prevotella intermedia isolated from subgingival plaque of chronic periodontitis patients. Settings and Design: Written informed consent was obtained from each subject enrolled in the study. The institutional ethics committee granted ethical clearance for the study. Materials and Methods: Twenty patients diagnosed with chronic periodontitis were included in the study. Using sterile curettes, pooled subgingival plaque samples were collected from the deepest sites of periodontal pockets. The collected samples were then transported in 1 mL of reduced transport fluid. The organisms were cultured and confirmed and were used for the minimum inhibitory concentration (MIC) procedure. Statistical Analysis: Mean of the MIC value obtained was calculated. Comparison between the antimicrobial activity of mycelium and spore against P. intermedia groups with respect to mean MIC value (in mcg/ml) was done by Mann–Whitney U test. Comparison of antimicrobial activity of mycelium and spores of G. lucidum against P. intermedia expressed by the MIC value at different concentrations was done by Chi-square test. Results: The mean MIC obtained for mycelium was 5.64 mcg/ml. The mean MIC obtained for spore was 3.62 mcg/ml. Results of the comparison of antimicrobial activity implied that the antimicrobial activity of mycelium and spore of G. lucidum was comparable. Conclusions: Both mycelium and spore of G. lucidum with its bioactivity could be used as an antimicrobial, in combination with conventional therapy in periodontal disease.

Summary: An 11‐year‐old American Quarter Horse gelding was referred to the JT Vaughan Large Animal Teaching Hospital at Auburn University, College of Veterinary Medicine, for investigation of acute and severe right‐sided facial swelling and nasal discharge. Standing computed tomographic (CT) examination of the head identified severe soft tissue swelling surrounding the right mandible, emphysema within the soft tissues tracking along fascial planes and right‐sided caudal and rostral maxillary sinusitis. Using CT identification and ultrasound guidance, several targeted fasciotomies were created into the right masseter, cranial cervical musculature, supraorbital space and caudo‐medial aspect of the mandible. The right‐sided sinusitis was treated by right conchofrontal sinus trephination and lavage. Aerobic and anaerobic cultures obtained from the fasciotomy sites and conchofrontal sinus both yielded Prevotella intermedia and Peptostreptococcus asaccharolyticus. During hospitalisation, serial, standing CT examinations were performed for monitoring case progression and guiding further fasciotomies in the face of continued myonecrosis. Follow‐up CT performed at 1 month showed resolution of the emphysema and presence of chronic right ventral conchal sinusitis. The sinusitis was treated by fenestration of the right ventral concha with a diode laser via the nasal passage. [ABSTRACT FROM AUTHOR]

Objective: To determine the correlation between levels of methyl mercaptan (CH3SH) hydrogen sulfide (H2S), the proportion of Prevotella intermedia (Pi), and matrix metalloproteinase-8 (MMP-8) gene expression levels in periodontitis patients accompanied by halitosis. Material and Methods: Samples were obtained from gingival crevicular fluid (GCF) in the deepest pocket and by swabbing in the tongue coating area in patients with periodontitis presenting with halitosis (n = 23) and healthy subjects as controls (n = 7). The values of CH3SH and H2S were obtained using Oral Chroma. The proportion of Pi and MMP-8 expression levels were evaluated using PCR-RT. All the result was statistically analyzed using SPSS software. Results: The levels of CH3SH and H2S in participants with PD ≥ 6 mm showed a robust negative correlation with the proportion of P. intermedia in GCF and tongue coating. No statistically significant association was detected between CH3SH and H2S levels and MMP-8 expression levels (p>0.05). Conclusion: There is no association between CH3SH and H2S levels, the proportion of P. intermedia, and MMP-8 expression in patients with periodontitis accompanied by halitosis. [ABSTRACT FROM AUTHOR]

Bacterial biofilms formed on the surface of tissues and biomaterials are major causes of chronic infections in humans. Among them, Porphyromonas gingivalis (P. gingivalis) and Prevotella intermedia (P. intermedia) are anaerobic pathogens causing dental infections associated with periodontitis. In this study, we evaluated the killing effect and underlying mechanisms of direct current (DC) as an antimicrobial method in vitro. We chose P. gingivalis and P. intermedia in different states to make comparisons of the killing effect of DC. By viable bacteria counting, fluorescent live/dead staining, reactive oxygen species (ROS) assay, addition of ROS scavenger DMTU and mRNA expression assay of ROS scavenging genes, the role of ROS in the killing effect was explored. The planktonic and biofilm states of two bacteria could be effectively killed by low-intensity DC. For the killing effect of 1000 μA DC, there were significant differences whether on planktonic P. gingivalis and P. intermedia (mean killing values: 2.40 vs 2.62 log 10 CFU/mL) or on biofilm state of those (mean killing values: 0.63 vs 0.98 log 10 CFU/mL). 1000 μA DC greatly induced ROS production and the mRNA expression of ROS scavenging genes. DMTU could partially decrease the killing values of DC and downregulate corresponding gene's expression. 1000 μA DC can kill P. gingivalis and P. intermedia in two states by promoting overproduction of ROS, and P. intermedia is more sensitive to DC than P. gingivalis. These findings indicate low-intensity DC may be a promising approach in treating periodontal infections. [ABSTRACT FROM AUTHOR]