Clinical diagnosis of hyposalivation in hospitalized patients

Objective

The aim of this study was to evaluate the effectiveness of clinical criteria for the diagnosis of hyposalivation in hospitalized patients.

Material and Methods

A clinical study was carried out on 145 subjects (48 males; 97 females; aged 20 to 90 years). Each subject was clinically examined, in the morning and in the afternoon, along 1 day. A focused anamnesis allowed identifying symptoms of hyposalivation, like xerostomia complaints (considered as a reference symptom), chewing difficulty, dysphagia and increased frequency of liquid intake. Afterwards, dryness of the mucosa of the cheecks and floor of the mouth, as well as salivary secretion during parotid gland stimulation were assessed during oral examination.

Results

Results obtained with Chi-square tests showed that 71 patients (48.9%) presented xerostomia complaints, with a significant correlation with all hyposalivation symptoms (p<0.05). Furthermore, xerostomia was also significantly correlated with all data obtained during oral examination in both periods of evaluation (p<0.05).

Conclusion

Clinical diagnosis of hyposalivation in hospitalized patients is feasible and can provide an immediate and appropriate therapy avoiding further problems and improving their quality of life.     

Caffeine increases the expression of cystatin SN in human submandibular acinar-like HSG cells

ObjectiveThe study aimed at evaluating in vitro the effect of caffeine on expression of cystatin SN, a potential marker of sensitivity to bitterness in humans.MethodsDifferentiation of human submandibular gland (HSG) cells was induced by culturing cells on Matrigel. Caffeine cytotoxicity was assessed over 3 days by the Resazurin test. Finally, effects of 5, 50 and 100 μM caffeine exposure on cystatin SN expression were explored over 3 days by ELISA.ResultsAt concentrations relevant to human adult plasma levels (5, 50 and 100 μM), caffeine did not affect cell viability whether cells were differentiated or not. Cystatin SN levels were overall higher in differentiated cells and increased with time in both conditions. There was a significant (p < 0.001) effect of caffeine on cystatin SN expression specifically in differentiated cells.ConclusionsThe HSG cell line proved to be a relevant tool to study in vitro the effect of caffeine at concentrations consistent with dietary intake in human subjects. The results suggest that salivary cystatin SN abundance may depend on caffeine intake, with possible consequences on taste sensitivity.     

Growth hormone and cortisol in serum and saliva

Salivary diagnosis is a developing area in clinical chemistry and dentistry. Cortisol analyses from saliva have been used in pediatric practice and as doping tests. Growth hormone (hGH), also a stress hormone, has not been analyzed from saliva. We studied the serum and saliva of 51 healthy subjects. The samples were taken at 8:00 in the morning after 12 h fasting. Cortisol concentrations were analyzed using RIA. An immunoradiometric assay was applied for analyzing serum and salivary hGH. The validity of this method developed in our laboratory was found to be good. The results showed correlation of salivary cortisol with that of serum (r = 0.47, P &lt; 0.001). Salivary hGH concentrations were 1000-fold lower than the respective values in serum, but a clear correlation was found between salivary and serum hGH levels (r = 0.59, P &lt; 0.001).     

Influence of exposure time to saliva and antioxidant treatment on bond strength to enamel after tooth bleaching: an in situ study

Objectives

This study evaluated the influence of different exposure times to saliva in situ in comparison with an antioxidant treatment on composite resin bond strength to human enamel restored after tooth bleaching.

Material and Methods

Forty human teeth specimens measuring 5x5 mm were prepared and randomly allocated into 5 groups with 8 specimens each: Gct (control group, restored on unbleached enamel); Gbl (restored immediately after bleaching); Gsa (bleached, treated with 10% sodium ascorbate gel for 60 min and restored); G7d (bleached, exposed to saliva in situ for 7 days and restored); and G14d (bleached, exposed to saliva in situ for 14 days and restored). Restored samples were cut into 0.8 mm² sticks that were tested in microtensile. Specimens were microscopically analyzed and failure modes were classified as adhesive, cohesive, or mixed. Pretest and cohesive failures were not considered in the statistical analysis, which was performed with one-way ANOVA and Tukey''s post-hoc test (α=0.05), with the dental specimen considered as the experimental unit.

Results

Mean bond strength results found for Gbl in comparison with Gct indicated that bleaching significantly reduced enamel adhesiveness (P<0.01). However, no statistically significant differences were found between Gct, Gsa and G7d (P>0.05). Bond strength found for G14d was significantly higher than for Gsa (P<0.01). Fractures modes were predominantly of a mixed type.

Conclusions

Bonding strength to bleached enamel was immediately restored with the application of sodium ascorbate and exposure to human saliva in situ for at least 7 days. Best results were obtained with exposure to human saliva in situ for 14 days. Treatment with sodium ascorbate gel for 60 min may be recommended in cases patients cannot wait for at least 7 days for adhesive techniques to be performed.     

Oral gram-positive bacterial DNA-based identification of saliva from highly degraded samples

Analyzing degraded evidence is an important challenge in forensic casework. Saliva remaining at a crime scene may deteriorate, due to various factors, making it difficult to identify. This study aims to clarify the efficacy of oral gram-positive and -negative bacterial DNA-based identification of saliva for analyzing highly degraded samples. Saliva samples were subjected to three different degradation treatments (heat denaturation: 40–80 °C in wet conditions; microbial decomposition: 1–5 days in humid soil; and ultraviolet (UV) irradiation: 0.01–1 J/cm²). We compared saliva markers’ detectability from the degraded samples—oral gram-positive bacterial DNA (Streptococcus salivarius and Streptococcus oralis), oral gram-negative bacterial DNA (Veillonella atypica and Prevotella maculosa) and salivary α-amylase. Oral bacterial DNA was detected using a melting curve analysis following real-time PCR. The efficacy of short tandem repeats (STR) and mitochondrial DNA (mtDNA) analyses were also compared. All oral bacterial DNA were detected with specific melting peaks from the heat-denatured samples, while neither catalytic nor immunochromatographic tests detected salivary α-amylase from the heat (80 °C) samples. The gram-positive bacterial DNA (S. salivarius and S. oralis) was detected from the microbial degradation (1–5 days) samples. In contrast, the gram-negative bacterial DNA (V. atypica and P. maculosa) and salivary α-amylase were not detected from samples treated for more than two days. UV exposure made bacterial DNA-based saliva identification difficult in a dose–dependent manner; however, UV irradiation did not influence protein-based saliva tests using salivary α-amylase as an indicator. As a result of STR and mtDNA typing, partial or null STR profiles were generated from the severely degraded (microbial (2–5 days) and UV (0.1–1 J/cm²) degradation) samples, but full mtDNA profiles were obtained from all degraded samples. The forensic applicability of bacterial DNA test evaluated, using mock case samples, indicates that the oral gram-positive bacterial DNA was more resistant to degradation than the other markers. We conclude that the oral gram-positive bacterial DNA-based examination could be useful for identifying saliva from severely environmentally-exposed forensic samples as well as mtDNA typing.     

脆弱拟杆菌DNA探针在临床上的应用

目的：在DNA水平上准确鉴定B.fragilis(脆弱拟杆菌）。方法：用脆弱拟杆菌种特异的探针pBF-15与分离株或标本进行DNA点杂交。结果：8株生化鉴定为脆弱拟杆菌的7株经DNA探针证实为脆弱拟杆菌；1株粪拟杆菌、1株吉氏拟杆菌和2株未能鉴定到种的拟杆菌经探针重新鉴定的为脆弱拟杆菌。用pBF-15直接检测了46例临床标本，其结果与常规法符合。结论：DNA探针鉴定脆弱拟杆菌或诊断脆弱拟杆菌感染比常规法准确、快速、简便。     

Evaluation of HPV-16 and HPV-18 specific antibody measurements in saliva collected in oral rinses and merocel® sponges

BackgroundCurrent Human papillomavirus (HPV) L1 VLP vaccines protect against HPV-16 and HPV-18-associated cancers, in females and males. Although correlates of protection have not been identified, HPV-specific antibodies at sites of infection are thought to be the main mechanism of protection afforded by vaccination. Oral sampling has gained increased attention as a potential alternative to serum in monitoring immunity to vaccination and understanding local immunity in oral cancers.MethodsSerum was collected via venipuncture, and saliva was collected via oral rinses and Merocel® sponges from healthy volunteers: 16 unvaccinated females, 6 females (ages 24–41) and 6 mid-adult aged males (ages 27–45) recipients of three doses of the HPV-16/18/6/11 vaccine (Gardasil®). Mid-adult male vaccine trial participants were compared to female participants. Samples were tested for anti-HPV-16 and anti-HPV-18 immunoglobulin G levels by an L1 virus-like particle-based enzyme-linked immunosorbent assay (ELISA).ResultsAll vaccinated participants had detectable serum anti-HPV-16 and anti-HPV-18 antibodies. Optimal standard concentration range and sample serial dilutions for oral rinses were determined. The standard curve was not affected by the type of solution examined. Reproducibility of HPV-16 and HPV-18 antibody titers in mouthwash (overall CV < 10%) or in Merocel® extraction buffer was robust (CV < 13%). Excellent assay linearity (R² > 0.9) was observed for sera spiked controls in both solutions. HPV-16 and HPV-18 specific antibodies were detectable in saliva from vaccine recipients, both in mouthwash and in Merocel® sponges but levels were several logs lower than those in serum.ConclusionsThis study confirms the application of HPV-16 and HPV-18 ELISAs currently used in sero-epidemiological studies of immunogenicity of HPV vaccines for use with oral samples. Oral samples may be a useful resource for the detection of HPV-16 and HPV-18-specific antibodies in saliva following vaccination.     

Detection and serotype distribution of Actinobacillus actinomycetemcomitans in cardiovascular specimens from Japanese patients

Actinobacillus actinomycetemcomitans, an important pathogen in periodontitis, has also been detected in cardiovascular tissues. Sixty heart valves were collected during valve replacement surgery from 60 patients (one from each), 10 were from patients with infective endocarditis (IE group) and 50 were from patients with other valvular diseases (non-IE group). In addition, 46 samples of aneurysmal tissue were taken from 46 patients with a thoracic or abdominal aneurysm (Aneurysm group, one from each). Dental plaque samples were taken from 54 of the patients, 31 in the IE and non-IE groups and 23 in the aneurysm group. First, the distribution of A. actinomycetemcomitans in all specimens was analysed using a polymerase chain reaction method, which resulted in a positive reaction in 33 (31.1%) of the cardiovascular specimens and 25 (46.3%) of the dental plaque samples. Next, using serotype-specific sets of primers, the serotype distribution of A. actinomycetemcomitans in the cardiovascular specimens and dental plaque samples was found to be significantly different compared to dental plaque samples from Japanese subjects reported previously.     

切胶免疫制备腮腺液高丰度蛋白多克隆抗体

目的 通过切胶免疫制备人腮腺液高丰度蛋白多克隆抗体,为下一步腮腺液高丰度蛋白单克隆抗体制备提供抗原解决方案.方法 将腮腺液经超滤法浓缩蛋白后,进行SDS-PAGE分析,切取分子量约为50-65kDa的高丰度蛋白条带,研磨后注射新西兰大耳白兔诱导产生免疫应答并制备兔抗人腮腺液高丰度蛋白多克隆抗体,并应用蛋白免疫印记(Western blot)等方法进行抗体鉴定.结果 成功制备和鉴定了人腮腺液高丰度蛋白多克隆抗体.结论 成功制备和鉴定了人腮腺液高丰度蛋白多克隆抗体,为下一步腮腺液高丰度蛋白单克隆抗体制备及唾液蛋白质组学研究奠定基础.