Protein Folding Activity of the Ribosome (PFAR) –– A Target for Antiprion Compounds

Prion diseases are fatal neurodegenerative diseases affecting mammals. Prions are misfolded amyloid aggregates of the prion protein (PrP), which form when the alpha helical, soluble form of PrP converts to an aggregation-prone, beta sheet form. Thus, prions originate as protein folding problems. The discovery of yeast prion(s) and the development of a red-/white-colony based assay facilitated safe and high-throughput screening of antiprion compounds. With this assay three antiprion compounds; 6-aminophenanthridine (6AP), guanabenz acetate (GA), and imiquimod (IQ) have been identified. Biochemical and genetic studies reveal that these compounds target ribosomal RNA (rRNA) and inhibit specifically the protein folding activity of the ribosome (PFAR). The domain V of the 23S/25S/28S rRNA of the large ribosomal subunit constitutes the active site for PFAR. 6AP and GA inhibit PFAR by competition with the protein substrates for the common binding sites on the domain V rRNA. PFAR inhibition by these antiprion compounds opens up new possibilities for understanding prion formation, propagation and the role of the ribosome therein. In this review, we summarize and analyze the correlation between PFAR and prion processes using the antiprion compounds as tools.     

UPLC-Q-TOF-MS对桑白皮总黄酮提取物的快速分析

目的:对桑白皮总黄酮提取物中的各化合物进行快速的分离和鉴定,并对其主要的裂解碎片和二级裂解规律进行探讨。方法:采用超高效液相串联四级杆飞行时间质谱联用技术(UPLC-Q-TOF-MS),在电喷雾离子源(ESI)负离子模式下进行分离和检测;使用Waters公司BRH分析柱(2.1 mm×50 mm,1.7μm);以0.1%甲酸水为流动相A,乙腈为流动相B,梯度洗脱;流速0.3 m L·min~(-1);柱温为25℃。结果:基于UPLC-Q-TOF-MS技术分离所得精确相对分子质量,结合二级质谱裂解信息,使用Masslynx 4.1软件对数据进行统计和分析,并参考Sci Finder数据库(美国化学学会在线数据库学术版),桑白皮总黄酮提取物中共鉴定出13种黄酮成分,其中有7个Diels-Alder加合物,主要代表物为桑根酮C(sanggenon C),桑根酮D(sanggenon D),6种异戊烯基黄酮类化合物,代表物为桑辛素(morusin)。结论:通过超高效液相的分离,首次发现了桑白皮总黄酮提取物中的化合物在ESI源中脱去H_2和H_2O的现象,并对其二级裂解碎片的规律进行探讨,同时也为桑白皮总黄酮提取物的成分鉴定提供了一个快速、简便、可靠的方法。     

牙龈卟啉单胞菌膜泡诱导牙龈上皮细胞炎性反应的体外研究

目的建立牙龈卟啉单胞菌膜泡诱导牙龈上皮细胞炎性反应的体外模型，探讨牙龈卟啉单胞菌在牙周炎中的致病作用。方法用酶联免疫吸附法检测牙龈卟啉单胞菌膜泡对牙龈上皮细胞前列腺素E2（prostaglandin E2，PGE2）分泌的影响，以实时反转录聚合酶链反应法检测牙龈卟啉单胞菌膜泡对牙龈上皮细胞环氧化物酶（cyclooxygenase，COX）-2和白细胞介素（interleukin，IL）-6、IL-8基因表达的作用。结果牙龈卟啉单胞菌膜泡浓度依赖性地促进了牙龈上皮细胞PGE2的分泌，并使COX-2、IL-6、IL-8的mRNA表达水平显著上调。结论牙龈卟啉单胞菌膜泡诱导牙龈上皮细胞发生的细胞炎性反应，可能是牙周炎发生、发展的重要因素。     

感官分析法和便携式硫化物检测仪检测口臭的相关性研究

目的比较感官分析法(鼻测法)和便携式硫化物检测仪(商品名Halimeter)检测口臭之间的准确性和一致性。方法首先对2名嗅辨员进行嗅觉检测试验,以确定其进行感官分析评分的资质。选择86例符合试验条件的受试者,分别用感官分析法评价口臭程度,硫化物检测仪检测口腔挥发性硫化物(VSC)含量,使用SPSS11.0软件分析鼻测法得分均值和VSC水平的关系。结果2名嗅辨员一致性良好,Kappa值达到0.568,P<0.01。鼻测法得分均值与Halimeter检测所得硫化物峰值之间相关系数r=0.538,P<0.01;与硫化物稳定值之间相关系数r=0.546,P<0.01。结论鼻测法得分和Halimeter检测的口腔VSC含量之间存在显著相关关系,提示Halimeter可在一定程度上作为鼻测法的辅助手段,用以检测口臭。     

Insoluble zinc, cupric and tin pyrophosphates inhibit the formation of volatile sulphur compounds

Oral malodour is mainly a result of the production of volatile sulphur compounds (VSC). The present study was concerned with investigating the anti-VSC effect of insoluble pyrophosphates (PP) of zinc, copper(II) and tin(II). The hypothesis to be tested was that the sulphide anions produced when VSC are solubilized in water have a higher affinity for the respective metal ions than the PP anion. The anti-VSC effects of insoluble PP were compared with the corresponding soluble metal salts using three in vitro methods: saliva putrefaction; dialysis of a suspension of PP and saliva against water; and analysis of water containing hydrogen sulphide and methyl mercaptan gases, and gases in the headspace. The levels of VSC were analysed by gas chromatography in the first and third methods, and released metal ions were analysed by atomic absorption spectroscopy in the second. The results showed that: the insoluble metal PP inhibited VSC formation in saliva by 99-100%; under dialysis, only minute amounts of metal ions are released from the combination of PP and saliva; and the PP lost their metal cations in water containing dissolved gases and inhibited VSC formation. Hence, the results support the experimental hypothesis. Sulphide ions are obviously very strong ligands for these metal ions.     

Clinical effects of a new mouthrinse containing chlorhexidine,cetylpyridinium chloride and zinc-lactate on oral halitosis. A dual-center,double-blind placebo-controlled study

OBJECTIVES: The aim of this double-blind, parallel study was to test the clinical efficacy of a newly developed mouthrinse in the treatment of oral halitosis in patients without periodontitis. MATERIAL AND METHODS: Forty volunteers, recruited in two centers, participated in this study. Patients were selected on the basis of (1) halitosis of oral origin, (2) full-mouth organoleptic score>1, using an arbitrary 0-5 scale, (3) level of volatile sulfur compounds (VSC)>170 parts per billion (ppb) and (4) Winkel tongue coating index (WTCI)>4 (0-12). Intervention included gargling with a mouthrinse containing chlorhexidine (0.05%), cetylpyridinium chloride (0.05%) and zinc-lactate (0.14%) or with a placebo mouthrinse without active ingredients. At days 0 and 14 clinical variables were assessed in order of performance: (1) organoleptic assessments, (2) levels of VSC, and (3) WTCI. RESULTS: Treatment with the active mouthrinse resulted in a significant mean reduction in the organoleptic score from 2.8 to 1.5 (p<0.005). In the placebo group, no significant reduction in the mean organoleptic score occurred. Consequently, this resulted, after 2 weeks, in a greater change of the organoleptic scores in the test group in comparison to the placebo group (p<0.005). The mean VSC scores were reduced from 292 to 172 ppb in the test group (p<0.005), whereas no reduction was observed in the placebo group. At the 2-week examination, the mean change of the VSC scores in the test group was significantly greater than the mean change in the placebo group (p<0.005). Neither in the test nor in the placebo group a significant reduction in tongue coating was observed. CONCLUSIONS: In conclusion, the tested mouthrinse is effective in the treatment of oral halitosis.     

Subjective patients' opinion and evaluation of halitosis using halimeter and organoleptic scores

Objectives:  The aim of the study was to evaluate the level of interest in fetor ex ore among respondents, patients of the Department of Conservative Dentistry in Warsaw.
Materials and Methods:  A questionnaire was completed by 295 patients, 202 females and 93 males aged 18–74 years (average 38.73 years). Each person was also examined for organoleptic score and volatile sulfur compounds (VSC) by Halimeter. Halitosis was diagnosed if the average level of VSC was ≥125 ppb and the organoleptic measurement using a 0–5 point scale was ≥2. Statistical analysis was performed using chi-square test.
Results:  Incidence of halitosis was greatest in age ranges 25–34 years (29.68%) and 45–54 years (24.52%). Sixty-eight (43.87%) persons with diagnosed halitosis frequently reported having a problem with bad breath and only 5.81% persons with halitosis did not notice this problem ( P  < 0.001). Among 54 healthy persons three (5.56%) reported having a problem with halitosis ( P  < 0.001).
Conclusion:  A statistically significant correlation was found between clinical organoleptic diagnosis of halitosis and VSC level by halimeter. Subjective patients' opinion correlated well with objective evaluation of halitosis.     

Combined effect of zinc ions and cationic antibacterial agents on intraoral volatile sulphur compounds (VSC)

OBJECTIVE: Volatile sulphur compounds (VSC) are major components of oral malodour. As both zinc ions and cationic antibacterial agents inhibit the formation of oral VSC, this study aimed to determine whether these agents combined have synergistic anti-VSC actions. METHODS: Baseline oral VSC measurements of mouth air from 10 volunteers following cysteine rinsing (6mM, pH 7.2) were obtained using gas chromatography (GC). Subjects rinsed for 1 min with 10ml of the test solutions, 0.3% zinc acetate (Zn), 0.025% chlorhexidine (CHX), 0.025% cetyl pyridinium (CPC), and the combinations Zn+CHX and Zn+CPC. Cysteine rinses were repeated at 1h, 2h and 3h and VSC measurements recorded. Three subjects rinsed with the Zn+CHX combination and fasted for 9h, undergoing cysteine rinses and VSC measurements at 3h intervals. 10 microl of the test solutions were also added to 1ml aliquots of human whole saliva (n=8). Following incubation at 37 degrees C for 24h VSC levels in the saliva headspace were measured by GC. Inhibition of VSC formation and the fractional inhibitory index indicating synergy were calculated. RESULTS: Zn+CHX mouthrinse had a synergistic anti-VSC effect, and was effective for at least 9h. Zn+CPC mouthrinse was less effective. Both combinations showed a synergistic inhibiting effect in-vitro. CONCLUSIONS: Synergy between Zn and the antibacterial agents confirms different mechanisms of operation.     

Association between oral malodor and adult periodontitis: a review

BACKGROUND: Bad breath has a significant impact on our daily social life to those who suffer from it. The majority of bad breath originates within the oral cavity. However, it is also possible that it can come from other sources such as gastric-intestine imbalance. The term "oral malodor" is used to describe a foul or offensive odor emanating from the oral cavity, in which proteolysis, metabolic products of the desquamating cell, and bacterial putrefaction are involved. Recent evidence has demonstrated a link between oral malodor and adult periodontitis. The process of developing bad breath is similar to that noted in the progression of gingivitis/periodontitis. Oral malodor is mainly attributed to volatile sulfur compounds (VSC) such as hydrogen sulfide, methyl mercaptan and dimethyl sulfide. The primary causative microbes are gram-negative, anaerobic bacteria that are similar to the bacteria causing periodontitis. These bacteria produce the VSC by metabolizing different cells/tissues (i.e., epithelial cells, leukocytes, etc.) located in saliva, dental plaque, and gingival crevicular fluid. Tongue surface is composed of blood components, nutrients, large amounts of desquamated epithelial cells and bacteria, suggesting that it has the proteolytic and putrefactive capacity to produce VSC. One of the challenges in dealing with oral malodor is to identify a reliable test for detecting bad breath. AIMS: The purposes of this review article were: (1) to correlate the relationship between oral malodor and adult periodontitis; (2) to analyze current malodor tests and discuss available treatment regimens.