中国林蛙蝌蚪的口器发育

采用扫描电镜和组织学技术观察了中国林蛙(Rana chensinensis)蝌蚪发育过程中口器外部形态结构的变化,以及中国林蛙蝌蚪口器内部结构特征.结果表明,在口器发育的初期,角质颌最先出现,接着出现唇乳突以及唇齿;在变态高峰期(G4l～G42),口器结构如唇齿、角质颌和唇乳突则是按以下顺序消失的,即唇齿最先消失,其次...     

Characterization of the Oral Fungal Microbiome (Mycobiome) in Healthy Individuals

The oral microbiome–organisms residing in the oral cavity and their collective genome–are critical components of health and disease. The fungal component of the oral microbiota has not been characterized. In this study, we used a novel multitag pyrosequencing approach to characterize fungi present in the oral cavity of 20 healthy individuals, using the pan-fungal internal transcribed spacer (ITS) primers. Our results revealed the “basal” oral mycobiome profile of the enrolled individuals, and showed that across all the samples studied, the oral cavity contained 74 culturable and 11 non-culturable fungal genera. Among these genera, 39 were present in only one person, 16 genera were present in two participants, and 5 genera were present in three people, while 15 genera (including non-culturable organisms) were present in ≥4 (20%) participants. Candida species were the most frequent (isolated from 75% of participants), followed by Cladosporium (65%), Aureobasidium, Saccharomycetales (50% for both), Aspergillus (35%), Fusarium (30%), and Cryptococcus (20%). Four of these predominant genera are known to be pathogenic in humans. The low-abundance genera may represent environmental fungi present in the oral cavity and could simply be spores inhaled from the air or material ingested with food. Among the culturable genera, 61 were represented by one species each, while 13 genera comprised between 2 and 6 different species; the total number of species identified were 101. The number of species in the oral cavity of each individual ranged between 9 and 23. Principal component (PCO) analysis of the obtained data set followed by sample clustering and UniFrac analysis revealed that White males and Asian males clustered differently from each other, whereas both Asian and White females clustered together. This is the first study that identified the “basal mycobiome” of healthy individuals, and provides the basis for a detailed characterization of the oral mycobiome in health and disease.     

Antibacterial Activity and Inhibition of Adherence of Streptococcus mutans by Propolis Electrospun Fibers

Mouth-dissolving fibers with antibacterial activity for the oral cavity were prepared by an electrospinning technique. Propolis extract was used as an active ingredient and polyvinylpyrrolidone (PVP) K90 as the polymer matrix. The morphology and diameter of the fibers were characterized by scanning electron microscopy. Antibacterial activity against Streptococcus mutans and the inhibition of S. mutans adhesion on a smooth glass surface during the biofilm formation were tested. Propolis, 5% (w/v), was combined with a PVP K90 solution, 8% (w/v), with or without Tween 80 including flavor additives and electrospun with an applied voltage of 15 kV. Uniform and smooth fibers of propolis-PVP K90 were obtained. The results showed that electrospun fibers with propolis extract can dissolve and release the propolis in water. Propolis-PVP electrospun fibers showed better antibacterial activity by reduction of bacteria adhesion on a smooth glass surface when compared to some commercial mouthwash products. These results indicated the potential of electrospun fibers to be used as mouth-dissolving fibers for effective antibacterial activity in the oral cavity.KEY WORDS: antibacterial activity, electrospun fibers, inhibition of adherence, propolis, Streptococcus mutans     

Osteological development of the feeding apparatus in early stage larvae of the seabass,Lates calcarifer

The osteological development of the feeding apparatus was examined in early stage larvae of laboratory-reared seabass,Lates calcarifer. At initial mouth openings 40 hours after hatching, the larvae were equipped with the fundamental elements forming the oral cavity, such as the trabecular roof, the lower branchial and hyoid arches forming the floor, the quadrate and symplectic-hyomandibular cartilages making up the sides, and the maxilla and Meckel's cartilage bordering the jaws. The cleithrum appeared almost simultaneously. The mechanics of creating a negative pressure in the oral cavity, which results in a “sucking” mode of feeding, were elucidated from these elements. During a period from 50–60 to 100–110 hours after initial mouth opening (HAMO), new elements such as the premaxilla and jaw teeth appeared, and the ossification of existing elements started. The new elements apparently enabled the larvae to “grasp” food organisms, in addition to the already existing and increasing sucking ability, from 100–110 HAMO.     

Axenic Culture of a Candidate Division TM7 Bacterium from the Human Oral Cavity and Biofilm Interactions with Other Oral Bacteria

The diversity of bacterial species in the human oral cavity is well recognized, but a high proportion of them are presently uncultivable. Candidate division TM7 bacteria are almost always detected in metagenomic studies but have not yet been cultivated. In this paper, we identified candidate division TM7 bacterial phylotypes in mature plaque samples from around orthodontic bonds in subjects undergoing orthodontic treatment. Successive rounds of enrichment in laboratory media led to the isolation of a pure culture of one of these candidate division TM7 phylotypes. The bacteria formed filaments of 20 to 200 μm in length within agar plate colonies and in monospecies biofilms on salivary pellicle and exhibited some unusual morphological characteristics by transmission electron microscopy, including a trilaminated cell surface layer and dense cytoplasmic deposits. Proteomic analyses of cell wall protein extracts identified abundant polypeptides predicted from the TM7 partial genomic sequence. Pleiomorphic phenotypes were observed when the candidate division TM7 bacterium was grown in dual-species biofilms with representatives of six different oral bacterial genera. The TM7 bacterium formed long filaments in dual-species biofilm communities with Actinomyces oris or Fusobacterium nucleatum. However, the TM7 isolate grew as short rods or cocci in dual-species biofilms with Porphyromonas gingivalis, Prevotella intermedia, Parvimonas micra, or Streptococcus gordonii, forming notably robust biofilms with the latter two species. The ability to cultivate TM7 axenically should majorly advance understanding of the physiology, genetics, and virulence properties of this novel candidate division oral bacterium.     

Serous goblet cells: The protein secreting cells in the oral cavity of a catfish, Rita rita (Hamilton, 1822) (Bagridae,Siluriformes)

Serous goblet cells in the oral epithelium of Rita rita are characterized by the presence of distinct eosinophilic granules occupying large parts of the cytoplasm. In R. rita, a range of histochemical results reveal that these cells are involved in proteinaceous secretions, and thus likely contribute to various functions analogous to those of mammalian saliva. The secretions of these cells have also been associated with specific functions and are discussed in relation to their physiological importance with special reference to their roles in lubrication, alteration in viscosity, various functions of mucus such as handling, maneuvering and driving of food items toward the esophagus, maintaining taste sensitivity and protection of the oral epithelium. In addition, the serous goblet cells may also be considered as the primary defensive cell of the oral epithelium of R. rita. The results significantly add to very limited set of literature on the serous goblet cells and provide noteworthy information on the mucous secretions in the oral cavity of fish.     

Detection of Mycobacterium leprae in saliva and the evaluation of oral sensitivity in patients with leprosy

The aim of this study was to investigate sensitivity disorders in the oral cavity related to the presence of Mycobacterium leprae in the saliva of treatment-naïve patients with leprosy in the state of Amazonas, Brazil. A cross-sectional study was conducted involving 45 subjects with leprosy. The subjects were interviewed to evaluate the sensitivity of the oral cavity. For the detection of M. leprae, saliva and slit-skin smear samples were collected. The samples were analysed using a bacteriological index (BI) protocol and the real-time quantitative polymerase chain reaction (qPCR). The results indicated that 15 of the 45 (33.3%) subjects with leprosy showed decreased oral sensitivity, which confirmed the importance of the oral cavity sensitivity evaluation. There was not a direct relationship between the presence of M. leprae in saliva and changes in oral sensitivity. Positive saliva qPCR results from six (31.6%) of 19 paucibacillary (PB) patients suggested the possibility of a new site for sample collection. Positive results using these diagnostic techniques (BI, slit-skin smear and saliva qPCR) increased to 55.5%, thus opening the possibility of combining these different techniques to increase the rate of positive diagnoses, especially in PB patients.     

FOOD VACUOLE MEMBRANE GROWTH WITH MICROTUBULE-ASSOCIATED MEMBRANE TRANSPORT IN PARAMECIUM

Evidence from a morphological study of the oral apparatus of Paramecium caudatum using electron microscope techniques have shown the existence of an elaborate structural system which is apparently designed to recycle digestive-vacuole membrane. Disk-shaped vesicles are filtered out of the cytoplasm by a group of microtubular ribbons. The vesicles, after being transported to the cytostome-cytopharynx region in association with these ribbons, accumulate next to the cytopharynx before they become fused with the cytopharyngeal membrane. This fusion allows the nascent food vacuole to grow and increase its membrane surface area. The morphology of this cytostome-cytopharynx region is described in detail and illustrated with a three-dimensional drawing of a portion of this region and a clay sculpture of the oral apparatus of Paramecium. Evidence from the literature for the transformation of food vacuole membrane into disk-shaped vesicles both from condensing food vacuoles in the endoplasm and from egested food vacuoles at the cytoproct is presented. This transformation would complete a system of digestive vacuole membrane recycling.     

Oral Colonization of Fungi

Opportunistic microbes are able to exist as commensals or pathogens depending on local environmental conditions. The bacterial microbiome at mucosal sites (gut, oral and vaginal) has been well characterized but there has been less focus on the fungal component of the microbiome, the “mycobiome”, especially of the oral mucosa. Genomic characterization studies have shown that Candida species are the most prevalent fungal species in the mycobiomes of the murine gut and human oral cavity, with C. albicans being the most abundant fungal species in the oral cavity. In this review, we outline recent advances in the characterization of the oral mycobiome, how different Candida species colonize, invade and infect the oral cavity, and how epithelial surfaces play a key role in antifungal activity and discriminate between commensal and pathogenic Candida.     

PCR analysis of Actinobacillus actinomycetemcomitans,Porphyromonas gingivalis,Treponema denticola andFusobacterium nucleatum in middle ear effusion

Anaerobes contribute to the severity and chronicity of infections that occur in and around the oral cavity. One of the factors involved in the pathogenesis of otitis media with effusion (OME) is the retrograde movement of bacteria from the oropharynx into the middle ear cavity. OME is one of the most common causes of hearing loss in children. We have used a PCR-based method to identify Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola andFusobacterium nucleatum in 65 middle ear effusion (MEE) samples obtained from paediatric patients seen for myringotomy and tube placement. DNA was extracted from MEE samples and PCR was initially done with DNA extracts by using the universal primers within the 16S rRNA gene sequence common to all bacterial species. The positive samples were further assessed with four species-specific primers. With the universal primers, 27 of 65 samples (41.5%) showed positive reaction indicating the presence of bacterial DNA. F. nucleatum was present in 10 out of 27 PCR-positive samples (37%) while one sample was positive for both T. denticola and F. nucleatum (3.7%). A. actinomycetemcomitans and P. gingivalis were not detected in any of the samples. The results of this study suggest that oral bacterial species may also play a role in the aetiopathogenesis of paediatric MEE.     

Do Maternal Microbes Shape Newborn Oral Microbes?

Strong evidence suggests that the early composition of the oral microbiota of neonates plays an important role for the postnatal development of the oral health or immune system. However, the relationship between the maternal microbiome and the initial neonatal microbiome remains unclear. In this study, 25 pregnant women and their neonates were recruited, and the samples were collected from the maternal oral cavity, amniotic fluid, placenta and neonatal oral cavity. High-throughput sequencing of 16S rRNA was performed using the Illumina MiSeq platform to analyze the correlation with microbial community structure between the maternal and the neonatal oral cavity. The results indicated that the number of shared OTUs was up to 635 in four groups. The PCoA showed that there were certain similarities in the microbial community structure of the four groups. The dominant bacterial genera of the shared OTUs were consistent with human oral microbes, including Streptococcus, Fusobacterium and Prevotella. The results showed that there might be a correlation between the maternal and neonatal oral microbiome, through the amniotic fluid and placenta.Electronic supplementary materialThe online version of this article (10.1007/s12088-020-00901-7) contains supplementary material, which is available to authorized users.     

Influence of Oral Probiotic <Emphasis Type="Italic">Streptococcus salivarius</Emphasis> K12 on Ear and Oral Cavity Health in Humans: Systematic Review

Traditionally, probiotics are linked to the good health of the intestine and most clinical studies focus on that field. Evidence of oral probiotic use for ear and oral cavity disease prevention with impact on human health is limited. This work reviews existing studies and literature on Streptococcus salivarius K12 as an oral probiotic and effects of S. salivarius K12 on human ear and oral cavity human health. The studies were accessed via database searches: MEDLINE, PubMed, and Elsevier. The search included/focused on/encompassed publications from 2003 to 2016 with keywords related to K12 Streptococcus salivarius, bacteriocin-like inhibitory substances (BLIS) K12, probiotic K12 salivarius, and K12 probiotic health effects. Only a small amount of studies was identified: the total of 68 studies was identified, 35 of which were relevant after screening, and 9 were included in the final analysis. Very little literature is available about the association/correlation between/connection/interrelation of S. salivarius K12 with/and human ear and oral cavity health. S. salivarius K12 may have a role in reducing the occurrence and/or severity of secretory otitis media (SOM) and also in prevention of streptococcal and viral pharyngotonsillitis in children. Research highlights that S. salivarius K12 has shown promising results in treatment of halitosis, but data are still deficient. Further studies need to be initiated to improve understanding of the association of oral probiotic S. salivarius K12 with human ear and oral cavity health.     

Oral Porphyromonas gingivalis translocates to the liver and regulates hepatic glycogen synthesis through the Akt/GSK-3β signaling pathway

Periodontal diseases are common chronic inflammatory disorders that result in the destruction of tissues around teeth. Many clinical studies suggest that periodontal diseases are risk factors for insulin resistance and diabetic mellitus development. However, the molecular mechanisms by which periodontal diseases regulate the progress of diabetes mellitus remain unknown. In this study, we investigated whether Porphyromonas gingivalis (P.g.), a major pathogen of periodontal diseases, present in the oral cavity, moves to the liver and affects hepatic glycogen synthesis. SNAP26b-tagged P.g. (SNAP-P.g.) was introduced into the oral cavity to induce periodontal disease in 4-week old female Balb/c mice. SNAP-P.g. was detected in the liver extracted from SNAP-P.g.-treated mice using nested PCR analysis. High blood glucose levels tended to promote SNAP-P.g. translocation from the oral cavity to the liver in mice. Periodic acid-Schiff staining suggested that hepatic glycogen synthesis decreased in SNAP-P.g.-treated mice. SNAP-P.g. was also internalized into the human hepatoma cell line HepG2, and this attenuated the phosphorylation of insulin receptor substrate (IRS)-1, Akt and glycogen synthase kinase-3β induced by insulin. Insulin-induced glycogen synthesis was suppressed by SNAP-P.g. in HepG2 cells. Our results suggest that P.g. translocation from the oral cavity to the liver may contribute to the progress of diabetes mellitus by influencing hepatic glycogenesis.     

Taste attractiveness of different hydrobionts for roach Rutilus rutilus, bitterling Rhodeus sericeus amarus, and rainbow trout Oncorhynchus mykiss

Comparative study of the taste attractiveness of different aquatic invertebrates (daphnids Daphnia longispina and D. pulex, larvae of Chironomidae, whirligig Gyrinus marinus, water skaters Gerris spp.) and plants (duckweed Lemna minor, filamentous alga Cladophora sp.) for roach Rutilus rutilus, bitterling Rhodeus sericeus amarus, and rainbow trout Oncorhynchus mykiss has been performed. Roach’s taste preferences for agar-agar pellets, containing the aqueous extracts of the organisms under study, varies from maximum (daphnids, filamentous alga, larvae of chironomids) to minimum (duckweed) or it can be absent (whirligig). Different taste preferences to pellets containing the extracts of aquatic organisms has been also found in bitterling (daphnids, larvae of chironomids, water skaters) but not in rainbow trout (daphnids, water skaters). It has been suggested that whirligigs and water skaters do not contain any deterrent substances ensuring chemical defense from predators. It has been demonstrated that intraoral gustatory reception of food items follows two alternative behavioral stereotypes that are different from each other in the time during which food items are retained in the oral cavity and the number of manipulations (subsequent grasps of a food item) during the perception process. We discuss the importance of gustatory reception as the major sensory mechanism that ensures feeding selectivity of fish and decreases interspecific competition over food.     

In vitro evaluation of the biomedical properties of chitosan and quaternized chitosan for dental applications

The aim of this study was to evaluate the potential dental applications of chitosan (CS) and N-[1-hydroxy-3-(trimethylammonium)propyl]chitosan chloride (HTCC). HTCC was prepared by reacting CS with glycidyltrimethylammonium chloride (GTMAC). CS and HTCC were characterized by infrared (FITR) and ¹H NMR spectroscopy. The antibacterial activity of CS and HTCC against oral pathogens, their proliferation activity and effects on the ultrastructure of human periodontal ligament cells (HPDLCs) were investigated. The results indicated that four oral strains were susceptible to CS and HTCC with minimum inhibitory concentrations (MICs) ranging from 0.25 to 2.5 mg/mL. The in vitro 3-(4,5-dimethyl-2-thizolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay determined that CS at 2000, 1000, 100, and 50 μg/mL could stimulate the proliferation of HPDLCs. Instead, HTCC inhibited the proliferation at the same concentrations but accelerated the proliferation of HPDLCs at relatively low concentrations (10, 3, 1.5, 1, and 0.3 μg/mL). Transmission electron microscopy (TEM) observations revealed that the ultra-architecture of HPDLC was seriously destroyed by HTCC treatment at 1000 μg/mL. Taken together, these results contribute information necessary to enhance our understanding of CS and HTCC in the dental field.     

Adaptation and Probiotic Potential of Lactobacilli,Isolated from the Oral Cavity and Intestines of Healthy People

The present study shows that, from 300 Lactobacillus strains isolated from the oral cavity and large intestine of 600 healthy people, only 9 had high antagonistic activity against pathogens and opportunistic pathogens. All antagonistic strains of lactobacilli have been identified by 16S rRNA sequencing and assigned to four species: Lactobacillus fermentum, Lactobacillus rhamnosus, Lactobacillus plantarum, and Lactobacillus casei. In addition, these lactobacilli appeared to be nonpathogenic and had some probiotic potential: the strains produced lactic acid and bacteriocins, showed high sensitivity to broad-spectrum antibiotics, and were capable of forming biofilms in vitro. With the help of PCR and specific primers, the presence of genes for prebacteriocins in L. plantarum (plnEF, plnJ, plnN) and L. rhamnosus (LGG_02380 and LGG_02400) has been revealed. It was found that intestinal strains of lactobacilli were resistant to hydrochloric acid and bile. Lactobacilli isolated from the oral cavity were characterized by a high degree of adhesion, whereas intestinal strains were characterized by average adhesion. Both types of lactobacilli had medium to high rates of auto-aggregation and hydrophobicity and could coaggregate with pathogens and opportunistic pathogens. Additionally, the ability of the lactobacilli strains to produce gasotransmitters, CH₄, CO₂, C₂H₆, CO, and NH₃, has been revealed.     

Understanding the Oral Mucosal Absorption and Resulting Clinical Pharmacokinetics of Asenapine

Absorption of drugs from the oral cavity into the mucosal tissues is typically a fast event. Dissolved drugs partition into the mucosal membranes and within minutes will reach equilibrium with drug in solution in the oral cavity. However, this does not always equate to rapid drug appearance in the systemic circulation. This has been attributed to slow partitioning out of the mucosal tissues and into the systemic circulation. Based on information from literature, physicochemical properties of asenapine, and clinical data, we conclude that for sublingually administered asenapine, the exposure is primarily a function of rapid partitioning into the mucosal membranes. This is followed by slow partitioning out of the mucosal tissues and into the systemic circulation, leading to a T_max value of about 1 h. The bioavailability of asenapine at doses below the saturation solubility in the mouth does not change and is controlled primarily by mass transport equilibrium. At doses above the saturation solubility, the bioavailability becomes more dependent not only on the distribution equilibrium but also on contact time in the mouth because additional variables (e.g. dissolution rate of the drug) need to be accounted for. These explanations are consistent with oral cavity absorption models from the literature and can be used to accurately describe the clinical data for asenapine.KEY WORDS: asenapine, exposure, oral mucosal absorption, T_max     

Ellagitannins modulate the inflammatory response of human neutrophils ex vivo

BackgroundTannin-rich plant materials are commonly used in the traditional medicine as external anti-inflammatory, antioxidant and antimicrobial agents. Plant extracts containing significant quantities of tannins are often used in the prevention and treatment of oral cavity diseases such as periodontosis or gingivitis. The contribution of pure ellagitannins to the observed anti-inflammatory activity of tannin-rich remedies is still not resolved.PurposeThe aim of the present study the study was to establish if ellagitannins and their precursor – pentagalloylglucose (1) can modulate the inflammatory response of ex-vivo stimulated neutrophils.MethodsHuman neutrophils were isolated from the buffy coats obtained from healthy volunteers. Neutrophils were cultivated with or without tested compounds. The influence of ellagitannins and 1 on the production and release of pro-inflammatory factors such as elastase, reactive oxygen species, interleukin-8, tumour necrosis factor alpha (TNF-α) and metalloproteinase-9 was evaluated using ELISA sets or chemical methods. The effect on surface expression of toll like receptor 4 (TLR-4) and apoptosis was also checked using flow cytometry.ResultsThe results showed that ellgitannins modulate the inflammatory response of human neutrophils by the inhibition of production and release of chosen cytokines and pro-inflammatory enzymes. By the induction of TNF-α ellagitannins enhance neutrophil apoptosis, which is of interest in the case of chronic inflammation within oral cavity. Ellagitannins also decrease the surface expression of TLR-4 in activated neutrophils.ConclusionThe results support the traditional use of tannin-rich products in the prevention and treatment of oral cavity diseases. The present study proves the substantial contribution of ellagitannins to the anti-inflammatory activity of tannin-rich medicinal plant materials.     

Microtubules, Microfilaments, and Membranes in Phagocytosis: Structure and Function of the Oral Apparatus of the Ciliate Climacostomum virens

Climacostomum virens uses oral membranelles to drive suspended food particles into its buccal cavity. The cavity leads to a buccal tube which extends into the cell by as much as half a cell length. The inner end of this tube is delimited by a haplokinety (two rows of basal bodies). Internal to this zone is the cytostome and cytopharynx where food vacuoles form. The buccal tube is encircled by a ring of fibrous material, the cytostomal cord, in the region of the cytostome immediately below the haplokinety. Ribbons of postciliary microtubules extend from the kinetosomes of the haplokinety, attach to the cytopharyngeal membrane, and pass under the cytostomal cord. They become broader and expand into the cytoplasm. Cytopharyngeal vesicles pass between the microtubular ribbons and fuse with the cytopharyngeal membrane to generate membrane for forming food vacuoles. The cytopharyngeal vesicles contain a material which is secreted into the forming food vacuoles. Ciliates continue to feed after incubation in a medium containing cycloheximide, indicating that they draw on a pre-existing pool of membrane when forming the food vacuole.