Metaproteomics Characterizes the Human Gingival Crevicular Fluid Microbiome Function in Periodontitis.

Periodontitis is the leading cause of tooth loss in adults worldwide. The human proteome and metaproteome characterization of periodontitis is not clearly understood. Gingival crevicular fluid samples were collected from eight periodontitis and eight healthy subjects. Both the human and microbial proteins were characterized by liquid chromatography coupled with high-resolution mass spectrometry. A total of 570 human proteins were found differentially expressed, which were primarily associated with inflammatory response, cell death, cellular junction, and fatty acid metabolism. For the metaproteome, 51 genera were identified, and 10 genera were found highly expressed in periodontitis, while 11 genera were downregulated. The analysis showed that microbial proteins related to butyrate metabolism were upregulated in periodontitis cases. In particular, correlation analysis showed that the expression of host proteins related to inflammatory response, cell death, cellular junction, and lipid metabolism correlates with the alteration of metaproteins, which reflect the changes of molecular function during the occurrence of periodontitis. This study showed that the gingival crevicular fluid human proteome and metaproteome could reflect the characteristics of periodontitis. This might benefit the understanding of the periodontitis mechanism.

Mandibular Buccal Bifurcation Cyst: Report of Two Cases.

Mandibular buccal bifurcation cyst is a rare inflammatory odontogenic cyst. We reported two cases who complained of painful swelling of extraoral soft tissue. Intraoral examination revealed the partially erupted mandibular first molar. Cone beam computed tomography showed a well-defined cystic lesion surrounding the first molar. Histopathologic images showed the cyst wall was infiltrated by a large number of plasma cells, neutrophils and eosinophils, and lined with a thin layer of non-keratinized stratified squamous epithelium. Finally, the two patients were diagnosed as mandibular buccal bifurcation cyst and treated with cyst enucleation and curettage.

Three-dimensional analysis of mandible ramus morphology and transverse stability after intraoral vertical ramus osteotomy.

OBJECTIVES: The purpose of this study was to investigate short- and long-term postoperative changes of both morphology and transverse stability in mandibular ramus after intraoral vertical ramus osteotomy (IVRO) in patients with jaw deformity using three-dimensional (3D) orthognathic surgery planning treatment software for measurement of distances and angles. STUDY DESIGN: This retrospective study included consecutive patients with skeletal Class III malocclusion who had undergone intraoral vertical ramus osteotomy and computed tomography images before (T0), immediately after (T1), and 1 year after (T2) surgery. Reference points, reference lines and evaluation items were designated on the reconstructed 3D surface models to measure distances, angles and volume. The average values at T0, T1, T2 and time-dependent changes in variables were obtained. RESULTS: After surgery, the condylar length, ramal height, mandibular body length and mandibular ramus volume were significantly decreased (P < 0.01), while clinically insignificant change was observed from T1 to T2. The angular length was increased immediately after surgery (P < 0.05), but it was decreased 1 year after surgery (P < 0.05). Lateral ramal inclination showed significant increase after surgery (P < 0.05) and maintained at T2. CONCLUSION: Changes in the morphology of the mandibular ramus caused by IVRO do not obviously bring negative effect on facial appearance. Furthermore, despite position and angle of mandibular ramus changed after IVRO, good transverse stability was observed postoperatively. Therefore, IVRO technique can be safely used without compromising esthetic results.

A qualitative and quantitative analysis of the human gingival crevicular fluid proteome and metaproteome.

Gingival crevicular fluid (GCF) is an integral part of oral fluid that plays a special role in maintaining the structure of junctional epithelium and defending against bacterial infection. In this study, we comprehensively analysed the composition of the human GCF proteome and metaproteome simultaneously to obtain multidimensional information about GCF. A total of 3680 human proteins (2540 with at least two unique peptides) were identified in the normal GCF sample, and their functions were mainly associated with immune function and inflammation. Among these proteins, 1874 proteins could be quantified by the iBAQ algorithm, and their abundances spanned a dynamic range of six orders of magnitude. For the GCF metaproteome, a total of 3082 proteins and 69 genera were found. In addition, 16 genera were not identified by GCF metagenomic analysis. Compared to the saliva metaproteome, 32 genera were found to be in common. The protein quantitative analysis showed that the abundance of GCF metaproteome contributed to approximately 4.17% of the total GCF proteome. The top three most abundant genera were Fusobacterium, Corynebacterium, and Leptotrichia. The above data will be useful for future research on GCF-related diseases.

Opto-Thermophoretic Attraction, Trapping, and Dynamic Manipulation of Lipid Vesicles.

Lipid vesicles are important biological assemblies, which are critical to biological transport processes, and vesicles prepared in the lab are a workhorse for studies of drug delivery, protein unfolding, biomolecular interactions, compartmentalized chemistry, and stimuli-responsive sensing. The current method of using optical tweezers for holding lipid vesicles in place for single-vesicle studies suffers from limitations such as high optical power, rigorous optics, and small difference in the refractive indices of vesicles and water. Herein, we report the use of plasmonic heating to trap vesicles in a temperature gradient, allowing long-range attraction, parallel trapping, and dynamic manipulation. The capabilities and limitations with respect to thermal effects on vesicle structure and optical spectroscopy are discussed. This simple approach allows vesicle manipulation using down to 3 orders of magnitude lower optical power and at least an order of magnitude higher trapping stiffness per unit power than traditional optical tweezers while using a simple optical setup. In addition to the benefit provided by the relaxation of these technical constraints, this technique can complement optical tweezers to allow detailed studies on thermophoresis of optically trapped vesicles and effects of locally generated thermal gradients on the physical properties of lipid vesicles. Finally, the technique itself and the large-scale collection of vesicles have huge potential for future studies of vesicles relevant to detection of exosomes, lipid-raft formation, and other areas relevant to the life sciences.

A Comparative Proteomics Analysis of Five Body Fluids: Plasma, Urine, Cerebrospinal Fluid, Amniotic Fluid, and Saliva.

PURPOSE: Body fluid is considered a rich source of disease biomarkers. Proteins in many body fluids have potential clinical applications for disease diagnostic and prognostic predictions. EXPERIMENTAL DESIGN: To determine differences in the protein components and functional features of body fluids, a proteomic comparison of five body fluids (plasma, urine, cerebrospinal fluid, saliva, and amniotic fluid) was conducted by high-resolution mass spectrometry. RESULTS: A total of 4717 nonredundant proteins were identified, and the concentrations of 3433 proteins were estimated by an intensity-based algorithm quantitation method. Among them, 564 proteins were shared among the five body fluids, with common functions in the coagulation/prothrombin system and inflammatory response. A total of 36.7% of the proteins were detected in only one body fluid and were closely related to their adjacent tissues by function. The functional analysis of the remaining 2986 proteins showed that similar functions might be shared among different body fluids, which highlighted intimate connection in the body. CONCLUSIONS AND CLINICAL RELEVANCE: The quantitative comparative functional analysis indicated that body fluids might reflect the diverse functions of the whole body rather than the characteristics of their adjacent tissues. The above data might indicate the potential application of body fluids for biomarker discovery.

Comparative proteomic analysis of the influence of gender and acid stimulation on normal human saliva using LC/MS/MS.

PURPOSE: Human saliva is an important source for disease biomarker discovery. This study is to investigate the influence of gender and acid stimulation on the normal human salivary proteome. EXPERIMENTAL DESIGN: Unstimulated and acid-stimulated saliva samples from 5 males and 5 females were labeled with 4-plex iTRAQ and analyzed by 2-DLC MS/MS. By bioinformatics analysis the gender and acid stimulation related proteins were defined. According to protein annotation the important proteins were validated by multiple reaction monitor analysis. RESULTS: A total of 1770 proteins were identified, and 82 proteins in unstimulated saliva were found to be gender-specific, mainly associated with immune function, metabolism and inflammation. However, no gender-specific proteins were found in acid-stimulated saliva. In addition, 182 and 307 differential proteins were found to be acid stimulation-specific in male samples and female samples, respectively, mainly participated in the process of cellular movement, immune function and inflammatory response. Besides, it was found that acid stimulation caused more significant alteration and played a more important role in the human salivary proteome than gender. Gender-specific (IGHG2 and TIMP1) and acid stimulation (PERL, ENOA, ACTB, B4E022 and CALL3) related proteins were validated by MRM analysis. CONCLUSIONS AND CLINICAL RELEVANCE: The results indicate that gender differences exist in the unstimulated salivary proteome, and the influence of acid stimulation on the salivary proteome was more significant than that of gender. The above results may be helpful for salivary proteome research in the future.