改进Trizol法提取白色念珠菌总RNA初探

目的:比较Trizol法、改进Trizol法和超声粉碎法提取白色念珠菌总RNA产量、纯度和完整性的差异。方法:对生长至静止期的白色念珠菌标准株ATCC10231分别采用Trizol法、改进Trizol法和超声粉碎法提取总RNA,用紫外分光光度计测量其A260,A280的值,并进行琼脂糖凝胶电泳,对总RNA的产量、纯度和完整性进行比较。结果:三种方法提取白色念珠菌总RNA产量分别为1.904μg、7.208μg和5.000μg,A260/A280比值分别为1.896、1.415和1.612。电泳结果显示改进Trizol法提取的总RNA呈现28 s rRNA,18 s rRNA和5 s rRNA三条清晰的条带,而Trizol法和超声粉碎法提取的总RNA均有降解。结论:改进Trizol法提取的白色念珠菌总RNA产量及纯度均高于Trizol法和超声粉碎法,且完整性更好。该法优于Trizol法和超声粉碎法。     

IL-1alpha, IL-1ra and IL-8 are differentially induced by Candida in experimental oral candidiasis

OBJECTIVE: To investigate the expression of interleukin-1alpha (IL-1alpha), IL-1ra and IL-8 by the oral epithelium challenged by various Candida species. MATERIALS AND METHODS: In vitro candidiasis was induced by C. albicans wild type SC5314, its EFG1, CPH1 and secretory aspartyl proteinase (SAP) mutants and, ATCC isolates of C. albicans, C. tropicalis and C. dubliniensis using a reconstituted human oral epithelium (RHOE) model. IL-1alpha, IL-1ra and IL-8 levels in culture media were quantified by an enzyme-linked immunosorbent assay at 12, 24 and 48 h. Fungal invasion and IL-1ra expression in RHOE were detected by periodic acid-Schiff staining and immunohistochemistry. RESULTS: Overall, the invasive Candida induced relatively higher levels of IL-1alpha, IL-1ra and IL-8 in the culture media than the noninvasive isolates. IL-1alpha and IL-1ra levels induced by Candida with hyphal invasion were significantly higher (P < 0.05) than those induced by the isolates without hyphal invasion at 12, 24 and 48 h. Candida albicans SC5314 induced IL-1ra expression in RHOE at 12 and 24 h but not at 48 h consistent with its hyphal invasion; while the noninvasive mutants and non-albicans Candida induced IL-1ra expression at 48 h. CONCLUSIONS: The cytokine expression profiles in experimental oral candidiasis may be associated with the invasive potential of Candida.     

Presence of mutans streptococci and Candida spp. in dental plaque/dentine of carious teeth and early childhood caries

This study determined the presence of mutans streptococci and Candida spp. in supragingival dental plaque and infected dentine of caries-free children, with early childhood caries and caries. Pooled samples of dental plaque and infected dentine were collected from 56 children aged 1-5 years, which were divided into 3 groups: early childhood caries (ECC); caries and caries-free. Infected dentine was collected in ECC and caries groups to compare the frequency of these microorganisms in the collected sites. The samples were inoculated in SB20 and SA medium, for mutans streptococci and Candida spp., respectively, and incubated at 37 degrees C for 48 h. Colony growth was verified and the identification was performed by biochemical tests and CHROMagar Candida. Fisher's test or chi-square (chi(2)) were applied (p=0.05). The more prevalent species were S. mutans and Candida albicans in ECC (85.4% and 60.4%, respectively), independently of the sample site. S. mutans only was significantly associated with carious teeth, whether in early childhood caries or not. However, the frequency of C. albicans in ECC was higher when compared to caries and caries-free groups. There is a significant association between the presence of C. albicans and early childhood caries.     

白假丝酵母菌对人脐静脉内皮细胞株ECV304细胞增殖的诱导作用

目的 研究白假丝酵母菌(S.albicans)对人脐静脉内皮细胞株ECV304细胞增殖及细胞周期的影响.方法 体外培养ECV304,实验分为S.albicans上清液组、S.albicans灭活菌液组、上清液和灭活菌液混合组、对照组.采用MTT法、细胞计数法、倒置显微镜及流式细胞术分别观察各组对ECV304细胞增殖及细...     

Candida attachment to oral epithelium

Inflamed oral mucosa biopsies from patients with thrush and high candidal density were observed in a transmission electron microscope (TEM) using ultra-histochemical staining with ruthenium red for glycocalyx visualization. Fimbriae comprising the glycocalyx and enabling yeast adhesion to epithelial cells were clearly visualized by ruthenium red. All internalized portions of the yeast walls were devoid of glycocalyx, indicating that the growing tips of the hyphae mechanically penetrated the host cells. The attachment of Candida occurred in two ways: by fimbria-mediated adhesion enabling colonization of the epithelial surface, and by invasion of the superficial epithelial cells via hyphae. As the interaction between adhesin receptors and adhesins stimulates the production of proinflammatory cytokines, Candida adhesion itself is assumed to induce mucosal inflammation.     

Candidal carriage and plaque distribution before, during and after removable orthodontic appliance therapy

Candidal counts at 6 mucosal sites in 33 subjects were estimated by imprint culture before, during and after removable orthodontic appliance therapy. Additionally, dental plaque scores and salivary pH were measured at each stage. Whole mouth and site prevalence of candida rose significantly during therapy before falling to levels approximating initial values. A similar pattern was seen for candidal counts for the whole mouth and at individual mucosal sites, although post-treatment densities were usually lower than originally seen. The number of sites colonised by candida and salivary pH also increased significantly during treatment and fell significantly afterwards. Only the upper palatal plaque scores rose during therapy and most other dental sites showed a gradual decrease in value during the course of the study. This could be attributed to regular monitoring of the patient's oral hygiene procedures. This investigation demonstrated a direct relationship between the presence of an acrylic appliance, candida and low salivary pH levels. Furthermore, removable appliance therapy had a positive, though transient influence upon the prevalence and density of oral candidal carriage, suggesting that the appliance may initiate the carrier state.     

ANTIMICROBIAL ACTIVITY OF SODIUM HYPOCHLORITE ASSOCIATED WITH INTRACANAL MEDICATION FOR Candida albicans AND Enterococcus faecalis INOCULATED IN ROOT CANALS

Objective:

The purpose of this study was to evaluate the action of sodium hypochlorite (NaOCl) associated with an intracanal medication against Candida albicans and Enterococcus faecalis inoculated in root canals.

Material and Methods:

Thirty-six human single-rooted teeth with single root canals were used. The canals were contaminated with C. albicans and E. faecalis for 21 days and were then instrumented with 1% NaOCl. The roots were divided into 3 groups (n=12) according to the intracanal medication applied: calcium hydroxide paste, 2% chlorhexidine (CHX) gel, and 2% CHX gel associated with calcium hydroxide. The following collections were made from the root canals: a) initial sample (IS): 21 days after contamination (control), b) S1: after instrumentation, c) S2: 14 days after intracanal medication placement; S3: 7 days after intracanal medication removal. The results were analyzed statistically by the Kruskal-Wallis test at 5% significance level.

Results and Conclusions:

Both 1% NaOCl irrigation and the intracanal medications were effective in eliminating E. faecalis and C. albicans inoculated in root canals.     

Adherence in vitro of Candida albicans to plasma treated acrylic resin. Effect of plasma parameters, surface roughness and salivary pellicle

The adhesion of Candida albicans to surfaces is the prerequisite for occurrence of denture stomatitis.

Objective

Hence, this study investigated if surface modifications with plasma treatments could reduce the adherence of C. albicans to a denture base resin.

Methods

Specimens (n = 180) with roughened and smooth surfaces were made and divided into five groups: control—specimens were left untreated; experimental groups—specimens were submitted to plasma treatments to obtain surfaces with different hydrophobicity (Ar/50 W; ArO₂/70 W; AAt/130 W) or incorporation of fluorine (Ar/SF₆70 W). Contact angle measurements were performed immediately after the treatments and after immersion in water for 48 h. For each group, half of the specimens were incubated with saliva prior to the adhesion assay. The number of adherent yeasts was evaluated by XTT reduction method.

Results

For the experimental groups, there was significant change in the mean contact angle after 48 h of immersion in water. Groups ArO₂/70 W and ArSF₆/70 W showed significantly lower absorbance readings than the other groups, regardless the presence or absence of saliva and surface roughness.

Conclusions

Results demonstrated that ArO₂/70 W and ArSF₆/70 W plasma treatments showed promising potential for reducing the adherence of C. albicans to denture base resins.     

Hydrophobicity of graphene as a driving force for inhibiting biofilm formation of pathogenic bacteria and fungi

Objective

To evaluate the surface and wettability characteristics and the microbial biofilm interaction of graphene coating on titanium.

口腔链球菌对白假丝酵母菌的拮抗作用

目的 观测3种口腔链球菌对白假丝酵母菌的生长有无抑制作用.方法 采用直接点种法、反点种菌落计数法和液体共培养法观测口腔变异链球菌、血链球菌、唾液链球菌对白假丝酵母菌的拮抗作用.结果 1)直接点种法中各组均未观察到清晰的抑菌圈;2)唾液链球菌软琼脂上的白假丝酵母菌菌落数较对照组减少(P<0.05);3)活菌与白假丝酵母菌...