Tumor-derived Prevotella intermedia aggravates gastric cancer by enhancing Perilipin 3 expression.

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.

Current research progress on Prevotella intermedia and associated diseases.

Prevotella 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.

Effect of nitro-conjugated linoleic acid on the inflammatory response of murine macrophages activated with lipopolysaccharide derived from Prevotella intermedia.

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-1ß 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-1ß 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.

Case report: isolated prevotella intermedia causing intracranial infection detected using metagenomic next generation sequencing.

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.

Antibacterial Efficacy of Matricaria recutita Essential Oil against Porphyromonas gingivalis and Prevotella intermedia: In Vitro Study.

AIM: To evaluate in vitro the antibacterial efficacy of Matricaria recutita (chamomile) essential oil at 50 and 75% against Porphyromonas gingivalis ATCC 33277 and Prevotella intermedia ATCC 25611 at 24 and 48 hours. MATERIAL AND METHODS: The sample consisted of 80 discs and Mueller-Hinton Agar, the medium chosen for the culture. To determine the bacterial sensitivity, discs were placed in each Petri dish with concentrations of essential oil at 50 and 75%, distilled water and 0.12% chlorhexidine. Subsequently, the inhibition halos were measured in millimeters at 24 and 48 hours after culture, with the Kirby-Bauer disk diffusion method. RESULTS: In groups treated with Porphyromonas gingivalis, measurements at 24 and 48 hours yielded 22.14 ± 2.61 and 22.63 ± 2.67 mm for 0.12% chlorhexidine, 18.90 ± 0.41 and 19.22 ± 0.54 mm for 75% essential oil, and 15.55 ± 0.45 and 15.77 ± 0.46 mm for 50% essential oil, respectively. No statistically significant differences were observed among the groups (p > 0.05). CONCLUSION: No significant differences were found between the antibacterial efficacy of 0.12% chlorhexidine and 50 and 75% essential oil of Matricaria recutita on Porphyromonas gingivalis and Prevotella intermedia at 24 and 48 hours. CLINICAL SIGNIFICANCE: The study demonstrates that essential oil derived from Matricaria recutita may effectively combat bacteria associated with periodontal disease. This discovery has the potential to impact dental practice by introducing a natural treatment option. Further research is warranted to fully elucidate the clinical significance and potential applications of this finding.

Effectiveness of Diode (810 nm) Laser in Periodontal Parameters and Reduction of Subgingival Bacterial Load in Periodontitis Patients.

AIM: This split-mouth randomized trial (RCT) aimed to assess the effect of diode laser on the clinical parameters in patients with periodontitis, compare the results with scaling and root planing (SRP) alone, and assess the implications of diode laser (DL) on plaque bacteria. MATERIALS AND METHODS: Seventeen periodontitis patients were randomly assigned into two equal groups based on the therapy delivered. Group I (control site) received just SRP at baseline, while group II (test site) received both SRP and DL irradiation. For both groups, the clinical periodontal parameters probing pocket depth (PPD), and clinical attachment level (CAL) were measured at baseline, 30 days, and 90 days. Microbiological amount was also measured at baseline, 30, and 90 days after periodontal treatment. The amounts of Aggregatibacter actinomycetemcomitans (A.a), Prevotella intermedia (Pr. intermedia), and Porphyromonas gingivalis (P. gingivalis) were determined using real-time PCR probing with specific bacterial primers. RESULTS: In both groups, PPD and CAL showed statistically significant reductions at different time intervals (p < 0.05). No significant difference were observed in CAL values after 1 and 3 months in both test and control groups (p > 0.05). The mean values of the concentration of A.a, Pr. intermedia and P. gingivalis were lower in the case group as compared to the control group and the difference was statistically significant after 1 month (*p = 0.001). CLINICAL SIGNIFICANCE: According to this study, non-invasive laser treatment has the potential to improve clinical outcomes by lowering the quantity of A.a, Pr. intermedia and P. gingivalis. CONCLUSION: In both groups, a considerable decrease in the periodontal pathogens A.a, Pr. intermedia and P. gingivalis were discovered; however, the intergroup comparison was insignificant in relation to PD and CAL. The adjunctive treatment with diode laser showed better efficacy in ensuring a better periodontal treatment than SRP alone. How to cite this article: Abdullah LA, Hashim N, Rehman MM, et al. Effectiveness of Diode (810 nm) Laser in Periodontal Parameters and Reduction of Subgingival Bacterial Load in Periodontitis Patients. J Contemp Dent Pract 2023;24(12):1008-1015.

Enrichment of Prevotella intermedia in human colorectal cancer and its additive effects with Fusobacterium nucleatum on the malignant transformation of colorectal adenomas.

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.

Isoeugenol-functionalized nanogels inhibit peri-implantitis associated bacteria in vitro.

OBJECTIVES: Obligate and facultative anaerobic bacteria adhering to dental implants are a major cause for peri-implant inflammation, which, if left untreated, can lead to implant loss. Previously, our group developed a new route for the synthesis of isoeugenol-functionalized aqueous nanogels for implant coatings. METHODS: Here, the antimicrobial activity of several new nanogels differing in spacer length (n = 6, 9, 44), radius (60-200 nm), and amount of isoeugenol functional substance (1-20 mol%) was tested against the following peri-implantitis-associated species: Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella intermedia, Aggregatibacter actinomycetemcomitans, Escherichia coli, Actinomyces viscosus, Enterococcus faecalis, Staphylococcus aureus, Streptococcus oralis, S. parasanguinis, and the yeast Candida albicans. The minimal bactericidal concentration (MBC) and fungicidal concentration (MFC) were determined for each combination. In addition, transmission electron microscopy (TEM) and fluorescence microscopy after live-dead-staining (LD-S) were performed to visualize nanogel-microbe interactions. RESULTS: Two nanogels, NG9-3 and NG9-4 (colloids of 80-150 nm, with a spacer length of n = 9 and feeding between 5 and 10 mol% isoeugenol), had an inhibitory effect on all Gram-positive species and on P. gingivalis and P. intermedia with MBC ≥31.25 µg/ml. TEM and LD-S images showed that cellular adhesion and uptake of nanogels resulted in swelling, shedding, or even complete detachment of the cell wall and then to bursting (see graphical abstract). CONCLUSIONS: Functional nanogels can be used as building blocks in the design of bioactive coatings on implants to prevent infection and accelerate tissue regeneration, but the concentrations required are higher than for antibiotics.

Microbiological and molecular identification of the anaerobic component of the oral microbiota in patients with cancer of the oropharyngeal region.

A research objective - to study tumor tissues of primary and recurrent patients with cancer of the oropharyngeal region for the frequency of occurrence of four types of anaerobic periodontogens and their associations by two methods: real-time PCR and cultural. There is speculation that bacteria can influence the pathogenesis of cancer. A comparative assessment of the content of four anaerobic periodontogens (Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis, Treponema denticola) in the tumor tissue and in the healthy tissue of the oral mucosa of patients with oropharyngeal cancer was carried out. It was found that the identification of odontopathogens by the real-time PCR method is much more informative than the traditional culture method, with the exception of P. intermedia, for the identification of which the traditional culture method was more effective. In 33.3% of patients, both primary and secondary, the composition of microorganisms was the same in both healthy and tumor tissue. In 20% of primary patients and in 13.3% of repeat patients, no associations of microorganisms included in the study were found in healthy tissue. Associations of 4 bacteria were recorded only in tumor tissue in both primary and repeated patients, and in repeated patients - statistically significantly more often. In 53.3% of repeat patients, associations of 4 bacteria were recorded in tumor tissue, whereas in primary patients, only in one case. P. gingivalis from tumor tissue in repeat patients was statistically significantly more often than in primary patients. T. forsythensis in primary patients was found statistically significantly more often in healthy tissues than in repeat patients, in which T. forsythensis was found statistically significantly more often from tumor tissue than in healthy tissue). T. denticola in healthy tissue was detected in both primary and repeated patients in isolated cases. T. denticola in tumor tissue was found statistically significantly more frequently in both primary and repeated patients compared to healthy tissue. P. gingivalis, T. forsythensis, and T. denticola should perhaps be considered risk indicators indicating the level of significance of their associations with oropharyngeal cancer.

Prevotella intermedia produces two proteins homologous to Porphyromonas gingivalis HmuY but with different heme coordination mode.

As part of the infective process, Porphyromonas gingivalis must acquire heme which is indispensable for life and enables the microorganism to survive and multiply at the infection site. This oral pathogenic bacterium uses a newly discovered novel hmu heme uptake system with a leading role played by the HmuY hemophore-like protein, responsible for acquiring heme and increasing virulence of this periodontopathogen. We demonstrated that Prevotella intermedia produces two HmuY homologs, termed PinO and PinA. Both proteins were produced at higher mRNA and protein levels when the bacterium grew under low-iron/heme conditions. PinO and PinA bound heme, but preferentially under reducing conditions, and in a manner different from that of the P. gingivalis HmuY. The analysis of the three-dimensional structures confirmed differences between apo-PinO and apo-HmuY, mainly in the fold forming the heme-binding pocket. Instead of two histidine residues coordinating heme iron in P. gingivalis HmuY, PinO and PinA could use one methionine residue to fulfill this function, with potential support of additional methionine residue/s. The P. intermedia proteins sequestered heme only from the host albumin-heme complex under reducing conditions. Our findings suggest that HmuY-like family might comprise proteins subjected during evolution to significant diversification, resulting in different heme coordination modes. The newer data presented in this manuscript on HmuY homologs produced by P. intermedia sheds more light on the novel mechanism of heme uptake, could be helpful in discovering their biological function, and in developing novel therapeutic approaches.

In vivo pharmacodynamics of lascufloxacin and levofloxacin against Streptococcus pneumoniae and Prevotella intermedia in a pneumonia mixed-infection mouse model.

This study aimed to evaluate the antimicrobial activity of a new quinolone, lascufloxacin, for the treatment of complicated pneumonia caused by Streptococcus pneumoniae and Prevotella intermedia using a neutropenic mice pneumonia mixed-infection model. In this study, one S. pneumoniae and four P. intermedia isolates were utilized. Antimicrobial efficacy was calculated for each isolate as the reduction of the bacterial count comparatively to the non-treated mice (log10 colony forming units (cfu)/mL) obtained in the lungs of the treated mice after 24 h. Consequently, the bacterial densities of S. pneumoniae (KY-9) and P. intermedia (335) in the lungs of control animals were 8.20 ± 0.19 log10 cfu/mL and 5.26 ± 1.50 log10 cfu/mL, respectively. At human-simulated doses, lascufloxacin and levofloxacin showed high antimicrobial activities against not only S. pneumoniae (lascufloxacin: 1.88 ± 0.43 log10 cfu/mL, p < 0.001; levofloxacin 4.30 ± 0.75 log10 cfu/mL, p < 0.001), but also P. intermedia (lascufloxacin: 1.54 ± 0.57 log10 cfu/mL, p < 0.001; levofloxacin: 2.79 ± 0.55 log10 cfu/mL, p = 0.0102). Additionally, levofloxacin demonstrated attenuated antimicrobial efficacies against S. pneumoniae in the mixed-infection model compared with that in the single infection model. In contrast, lascufloxacin showed enhanced antimicrobial activities against S. pneumoniae and P. intermedia in the mixed-infection model. In conclusion, lascufloxacin resulted in enhanced efficacies against S. pneumoniae and P. intermedia, in both the single and mixed-infection models used. These data support the clinical utility of lascufloxacin for use against S. pneumoniae and P. intermedia in the treatment of pneumonia.

Removable denture is a risk indicator for peri-implantitis and facilitates expansion of specific periodontopathogens: a cross-sectional study.

BACKGROUND: The prevalence of peri-implantitis ranges between 7 and 38.4% depending on risk indicators such as smoking, diabetes mellitus, lack of periodontal maintenance program, and history or presence of periodontitis. Currently, the possible effect of the type of superstructure on peri-implant health is unclear. This cross-sectional study aims to investigate the influence of the superstructure on the prevalence of peri-implant mucositis, peri-implantitis and peri-implant dysbiosis. METHODS: During a 32-month recruitment period dental implants were assessed to diagnose healthy peri-implant tissues, mucositis or peri-implantitis. The study included 1097 implants in 196 patients. Out of all peri-implantitis cases 20 randomly chosen submucosal biofilms from implants with fixed denture (FD) originating from 13 patients and 11 biofilms from implants with removable dentures (RD) originating from 3 patients were studied for microbiome analysis. Composition of transcriptionally active biofilms was revealed by RNAseq. Metatranscriptomic profiles were created for thirty-one peri-implant biofilms suffering from peri-implantitis and microbiome changes associated with superstructure types were identified. RESULTS: 16.41% of the implants were diagnosed with peri-implantitis, 25.00% of implants with RD and 12.68% of implants with FD, respectively. Multivariate analysis showed a significant positive association on patient (p = < 0.001) and implant level (p = 0.03) between the prevalence of peri-implantitis and RD. Eight bacterial species were associated either with FD or RD by linear discriminant analysis effect size method. However, significant intergroup confounders (e.g. smoking) were present. CONCLUSIONS: Within the limitations of the present work, RDs appear to be a risk indicator for peri-implantitis and seem to facilitate expansion of specific periodontopathogens. Potential ecological and pathological consequences of shift in microbiome from RDs towards higher activity of Fusobacterium nucleatum subspecies animalis and Prevotella intermedia require further investigation.

Characterization of substrate specificity and novel autoprocessing mechanism of dipeptidase A from Prevotella intermedia.

Prevotella intermedia, a Gram-negative anaerobic rod, is frequently observed in subgingival polymicrobial biofilms from adults with chronic periodontitis. Peptidases in periodontopathic bacteria are considered to function as etiological reagents. Prevotella intermedia OMA14 cells abundantly express an unidentified cysteine peptidase specific for Arg-4-methycoumaryl-7-amide (MCA). BAU17746 (locus tag, PIOMA14_I_1238) and BAU18827 (locus tag, PIOMA14_II_0322) emerged as candidates of this peptidase from the substrate specificity and sequence similarity with C69-family Streptococcus gordonii Arg-aminopeptidase. The recombinant form of the former solely exhibited hydrolyzing activity toward Arg-MCA, and BAU17746 possesses a 26.6% amino acid identity with the C69-family Lactobacillus helveticus dipeptidase A. It was found that BAU17746 as well as L. helveticus dipeptidase A was a P1-position Arg-specific dipeptidase A, although the L. helveticus entity, a representative of the C69 family, had been reported to be specific for Leu and Phe. The full-length form of BAU17746 was intramolecularly processed to a mature form carrying the N-terminus of Cys15. In conclusion, the marked Arg-MCA-hydrolyzing activity in Pre. intermedia was mediated by BAU17746 belonging to the C69-family dipeptidase A, in which the mature form carries an essential cysteine at the N-terminus.

Analysis of Oral Microbiota Revealed High Abundance of Prevotella Intermedia in Gout Patients.

BACKGROUND/AIMS: Microbes reside in a number of body sites, including the oral cavity, and are associated with the progression of many systemic diseases. In this study, we aimed to investigate the effects of gout and hyperuricemia (HUA) on the composition of oral microbiomes. METHODS: Analysis of the oral microbiota from 12 gout patients, 11 HUA patients, and 19 healthy control subjects was performed using a deep sequencing approach, and validation of significant changes in Prevotella intermedia and Serratia marcescens in new patient cohorts was performed using quantitative PCR (qPCR). RESULTS: Our analysis indicated that both gout and HUA significantly altered the composition of the oral microbiome in patients. Patients with gout or HUA had significantly greater levels of salivary Prevotella intermedia but significantly lower levels of Serratia marcescens than healthy control subjects. CONCLUSION: We demonstrated the association between the oral microbiome and gout and HUA for the first time. In particular, 16S sequencing and qPCR analysis revealed significantly higher levels of oral Prevotella intermedia in gout/HUA patients, which suggests that these patients might be at risk for the development of periodontitis.

Chemical composition, cytotoxic effect and antimicrobial activity of Stachys koelzii Rech.f. essential oil against periodontal pathogen Prevotella intermedia.

Prevotella intermedia is associated with periodontal diseases and endodontic infections. Periodontitis can be suppressed by utilizing the antiseptics, which target the infectious bacteria. The member of Stachys sp. has been used traditionally in the form of decoction or infusion for management of infectious diseases. The subject of this article was to evaluate the chemical composition, antimicrobial and cytotoxic effect of Stachys koelzii essential oil and its main components against Prevotella intermedia. GC-FID and GC-MS analysis were used to determine the chemical composition. The antimicrobial effects of S. koelzii essential oil was evaluated by micro-broth dilution assay. Time kill curve assays, leakage of cytoplasmic materials and anti-biofilm effects were determined. Its cytotoxic effect was evaluated by MTT assay. Essential oil with main components of α-pinene, trans-caryophyllene and 1,8-cineole inhibited P. intermedia with MIC and MBC values of 0.1 and 0.2 mg/mL. Its biofilm formation was higher than α-pinene, followed by trans-caryophyllene and 1,8-cineole. Essential oil and its main components increased the leakage of cytoplasmic components. Essential oil showed cytotoxic effect on HeLa cell lines with IC50 0.06 mg/mL. The cytotoxic effect of α-pinene on healthy cell lines was higher than essential oil. S. koelzii essential oil can be used in mouthwash formulations and its efficacy should be evaluated in large clinical studies.

A chinese case of prevotella intermedia and streptococcus constellatus intracranial mixed infection.

Streptococcal Species is increasingly recognized as a potentially preventable emerging infection in human's brain with high prevalence around the world. Streptococcus constellatus is one of the most common pathogens. Meanwhile, anaerobic bacteria are the rare causes for intracranial infection. To date, intracranial mixed infection caused by Prevotella intermedia and Streptococcus constellatus has not been reported. We reported a Chinese case to raise the global awareness of severity of the intracranial mixed infection. Here, we illustrated the epidemiological risk factors, clinical manifestations and outcomes of the patient. For patients who suffer from exacerbated brain infection with fetid cerebrospinal fluid, early repeated imaging is urgently needed and empiric antibiotic therapy should consider anaerobic and aerobic bacteria in these situations.

Degradation of Incretins and Modulation of Blood Glucose Levels by Periodontopathic Bacterial Dipeptidyl Peptidase 4.

Severe periodontitis is known to aggravate diabetes mellitus, though molecular events related to that link have not been fully elucidated. Porphyromonas gingivalis, a major pathogen of periodontitis, expresses dipeptidyl peptidase 4 (DPP4), which is involved in regulation of blood glucose levels by cleaving incretins in humans. We examined the enzymatic characteristics of DPP4 from P. gingivalis as well as two other periodontopathic bacteria, Tannerella forsythia and Prevotella intermedia, and determined whether it is capable of regulating blood glucose levels. Cell-associated DPP4 activity was found in those microorganisms, which was effectively suppressed by inhibitors of human DPP4, and molecules sized 73 kDa in P. gingivalis, and 71 kDa in T. forsythia and P. intermedia were immunologically detected. The kcat/Km values of recombinant DPP4s ranged from 721 ± 55 to 1,283 ± 23 µM-1s-1 toward Gly-Pro-4-methylcoumaryl-7-amide (MCA), while those were much lower for His-Ala-MCA. Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) analysis showed His/Tyr-Ala dipeptide release from the N termini of incretins, glucagon-like peptide 1 (GLP-1) and glucose-dependent insulinotropic polypeptide, respectively, with the action of microbial DPP4. Moreover, intravenous injection of DPP4 into mice decreased plasma active GLP-1 and insulin levels, accompanied by a substantial elevation in blood glucose over the control after oral glucose administration. These results are the first to show that periodontopathic bacterial DPP4 is capable of modulating blood glucose levels the same as mammalian DPP4; thus, the incidence of periodontopathic bacteremia may exacerbate diabetes mellitus via molecular events of bacterial DPP4 activities.

Quantitative detection of periodontopathic bacteria in lower respiratory tract specimens by real-time PCR.

The presence of common periodontopathic bacteria, the Fusobacterium nucleatum-periodonticum-simiae group, Prevotella intermedia, and Porphyromonas gingivalis was determined from respiratory tract specimens of bacterial pneumonia by real-time PCR using universal and species-specific TaqMan probe/primer sets. 42 patients with infectious pneumonia and 45 patients without infectious pneumonia were retrospectively enrolled in clinical studies. Periodontopathic bacterial DNA was found in 57.1% cases of infectious pneumonia and 31.1% cases of noninfectious pulmonary disease. However, the proportion of periodontopathic bacterial DNA did not differ between the two groups, and the presence or proportion of periodontopathic bacterial DNA was not related to any clinical index of pneumonia. Only two pneumonia cases consisted of >30% Fusobacterium DNA, suggesting that Fusobacterium was the causal pathogen in these cases. Our findings suggest that the periodontopathic bacteria rarely proliferate and be etiological pathogen in lower airway tract. However, further study is necessary, focusing on the pathogenicity of F. nucleatum in pulmonary infectious disease.

Selective photoantisepsis.

BACKGROUND AND OBJECTIVE: Selective killing of pathogens by laser is possible due to the difference in absorption of photon energy by pathogens and host tissues. The optical properties of pathogenic microorganisms are used along with the known optical properties of soft tissues in calculations of the laser-induced thermal response of pathogen colonies embedded in a tissue model. The objective is to define the laser parameters that optimize pathogen destruction and depth of the bactericidal effect. MATERIALS AND METHODS: The virtual periodontium is a computational model of the optical and time-dependent thermal properties of infected periodontal tissues. The model simulates the periodontal procedure: Laser Sulcular Debridement.1 Virtual pathogen colonies are placed at different depths in the virtual periodontium to determine the depth for effective bactericidal effects given various laser parameters (wavelength, peak power, pulse duration, scan rate, fluence rate) and differences in pathogen sensitivities. RESULTS: Accumulated background heat from multiple passes increases the depth of the bactericidal effect. In visible and near-IR wavelengths the large difference in absorption between normal soft tissue and Porphyromonas gingivalis (Pg) and Prevotella intermedia (Pi) results in selective destruction. Diode laser (810 nm) efficacy and depth of the bactericidal effect are variable and dependent on hemin availability. Both pulsed-Nd:YAG and the 810 nm diode lasers achieve a 2-3 mm deep damage zone for pigmented Pg and Pi in soft tissue without surface damage (selective photoantisepsis). The model predicts no selectivity for the Er:YAG laser (2,940 nm). Depth of the bactericidal effect is highly dependent on pathogen absorption coefficient. Highly sensitive pathogens may be destroyed as deep as 5-6 mm in soft tissue. Short pulse durations enable confinement of the thermal event to the target. Temporal selectivity is achieved by adjusting pulse duration based on target size. CONCLUSION: The scatter-limited phototherapy model of the infected periodontium is applied to develop a proper dosimetry for selective photoantisepsis. Dosimetry planning is essential to the development of a new treatment modality. Lasers Surg. Med. 48:763-773, 2016. © 2016 Wiley Periodicals, Inc.

Accuracy of commercial kits and published primer pairs for the detection of periodontopathogens.

OBJECTIVES: Despite the input of microbiome research, a group of 20 bacteria continues to be the focus of periodontal diagnostics and therapy. The aim of this study was to compare three commercial kits and laboratory-developed primer pairs for effectiveness in detecting such periodontopathogens. MATERIALS AND METHODS: Fourteen bacterial mock communities, consisting of 16 randomly assembled bacterial strains, were used as reference standard for testing kits and primers. Extracted DNA from mock communities was analyzed by PCR in-house with specific primers and forwarded for analysis to the manufacturer's laboratory of each of the following kits: ParoCheck®Kit 20, micro-IDent®plus11, and Carpegen® Perio Diagnostik. RESULTS: The kits accurately detected Fusobacterium nucleatum, Prevotella intermedia/Prevotella nigrescens, Parvimonas micra, Aggregatibacter actinomycetemcomitans, Campylobacter rectus/showae, Streptococcus mitis, Streptococcus mutans, and Veillonella parvula. The in-house primers for F.nucleatum were highly specific to subtypes of the respective periopathogen. Other primers repeatedly detected oral pathogens not present in the mock communities, indicating reduced specificity. CONCLUSIONS: The commercial kits used in this study are reliable tools to support periodontal diagnostics. Whereas the detection profile of the kits is fixed at a general specificity level, the design of primers can be adjusted to differentiate between highly specific strains. In-house primers are more error-prone. Bacterial mock communities can be established as a reference standard for any similar testing. CLINICAL RELEVANCE: The tested kits render good results with selected bacterial species. Primers appear to be less useful for routine clinical diagnostics and of limited applicability in research. Basic information about the periodontopathogens identified in this study supports clinical decision-making.