Saliva profiles in children using heart failure medication: a pilot study.

AIM: To study the saliva profiles in children with severe heart disease taking heart failure medication compared with the saliva from healthy age and gender matched controls. STUDY DESIGN: Cross sectional case-control design. METHODS: Twenty-four age and gender matched pairs of children, mean age 12.0 years participated. Stimulated saliva was collected in a standardized way before lunchtime and the subjects were asked to refrain from all eating, drinking and tooth brushing 90 mins before sampling. Stimulated salivary secretion rate, buffering capacity, total salivary viable count of bacteria, mutans streptococci and lactobacilli, calcium, chloride, magnesium, potassium, sodium and salivary IgA were determined. RESULTS: There were 7 of the 24 children in the cardiac group who had secretions below 0.5 ml/min compared with no child in the control group (p<0.01). Lower [corrected] total viable counts of bacteria (TVC) were detected in the cardiac group 1.4x106 ± 1.2x107 vs. 2.7x106 ± 2.9x107 in the control group (p<0.05). Mutans streptococci (MS) in the cardiac group were 5.2x104 ± 1.5x105 vs. 8.1 x10³ ± 1.3x104 in the control group, (p>0.05) and MS ratio of TVC constituted 0.11±0.35 per cent compared to 0.01±0.02 per cent for the control group (p>0.05). STATISTICS: Continuous data were analysed by an analysis of variance (ANOVA) and categorical data by chi-square test. CONCLUSION: Reduced salivary secretion could be a caries risk factor in children taking heart failure medication.

Effect of long-term consumption of milk supplemented with probiotic lactobacilli and fluoride on dental caries and general health in preschool children: a cluster-randomized study.

The aim of this study was to evaluate the effect of milk supplemented with probiotic bacteria and fluoride on caries development and general health in preschool children. Children 1-5 years of age (n = 248) attending 14 day care centres with 27 units in northern Sweden entered the study. The centres were randomly assigned to two parallel groups: children in the intervention group were served 150 ml milk supplemented with Lactobacillus rhamnosus LB21 (10(7) CFU/ml) and 2.5 mg fluoride per litre for lunch while the control group received standard milk. The double-blind intervention lasted for 21 months (weekdays) and data were collected through clinical examinations and questionnaires. The primary outcome was caries increment and secondary outcomes were measures of general health. The dropout rate was 25%. The mean baseline caries experience was 0.5 dmfs in the intervention units and 0.6 in the control units and after 21 months 0.9 and 2.2 (p < 0.05). The number of days with sick leave was similar in both groups but the children of the intervention units displayed 60% fewer days with antibiotic therapy (mean 1.9 vs. 4.7 days) and 50% less days with otitis media (0.5 vs. 1.0) (p > 0.05). In children who had participated during the whole 21-month intervention, fewer days with otitis media were reported (0.4 vs. 1.3 days, p < 0.05). No serious side effects were reported. It is concluded that daily consumption of milk containing probiotic bacteria and fluoride reduced caries in preschool children with a prevented fraction of 75%. Additional beneficial health effects were evident.

Sugar fermentation in probiotic bacteria--an in vitro study.

INTRODUCTION: Food supplemented with probiotic bacteria is a rapidly growing sector of the market. The aim of the present study was to evaluate and compare the acid production of selected probiotic strains available in commercial products. METHODS: Six Lactobacillus strains (Lactobacillus plantarum 299v and 931; Lactobacillus rhamnosus GG and LB21; Lactobacillus paracasei subsp. paracasei F19, and Lactobacillus reuteri PTA 5289) were cultivated at 37 degrees C in an anaerobic atmosphere on Man, Rogosa, Shape (MRS) agar for 48 h or MRS broth for 16 h. After centrifugation, the cells were washed and resuspended in sterile phosphate-buffered saline and immediately subjected to a fermentation assay with 12 different carbohydrates (nine sugars and three sugar alcohols) in microtiter plates with a pH indicator. The plates were examined for color changes after 24, 48, and 72 h of incubation under aerobic and anaerobic conditions. Three scores were used: negative (pH > 6.8); weak (pH 5.2-6.8), and positive (pH < 5.2). The strains were characterized with the API 50 CH system to confirm their identity. RESULTS: L. plantarum fermented all the sugars except for melibiose, raffinose, and xylitol. Both L. rhamnosus strains were generally less active although L. rhamnosus GG was slightly more active than strain LB21 in the 5% CO(2) setting. The latter strain exhibited negative reactions for sucrose, maltose, arabinose, and sorbitol under anaerobic conditions. The assays with L. paracasei and L. reuteri had negative or weak reactions for all tested sugars under both aerobic and anaerobic conditions. CONCLUSION: The metabolic capacity to form acid from dietary sugars differed significantly between the various probiotic strains.

Influence of a low xylitol-dose on mutans streptococci colonisation and caries development in preschool children.

AIM: To evaluate the effect of xylitol-containing tablets on mutans streptococci colonisation and caries development in preschool children. STUDY DESIGN: Randomised single-blind prospective design. METHODS: The material consisted of 132 healthy 2-year-old children, 71 boys and 61 girls and they were assigned to a xylitol tablet (test) group or a non-intervention control group. The mean age was 2 years + 1 month in both groups. The drop-out rate was 10.6% during the 2-year trial. The test group was given 1-2 xylitol tablets (0.5-1g) per day during 1.5 years. Mutans streptococci (MS) enumeration was performed at baseline and semi-annually in the children and at baseline or shortly after in the mothers with a chair-side technique. Caries prevalence was scored at baseline and the age of 4 years. RESULTS: No statistically significant differences in MS colonisation were disclosed between the test and control groups at baseline or any of the designated follow-ups. A statistically significant positive relationship was found between the maternal salivary MS levels and the colonisation of the children in the control group at 2.5 years, 3 years and 3.5 years (r=0.39, r=0.35; r=0.30; p<0.01, p<0.01 and p<0.05) but not in the xylitol tablet group (p<0.05). The mean caries prevalence was lower in the test group compared with the control group at 4 years of age (dmfs 0.38 +/-1.05 vs. 0.80 +/-2.60) but the difference was not statistically significant. CONCLUSION: The findings do not support a low-dose xylitol tablet program for caries prevention in preschool children.

Xylitol concentration in saliva and dental plaque after use of various xylitol-containing products.

The study consisted of two sets of experiments, one in saliva and one in dental plaque. The xylitol concentration in saliva was determined enzymatically in 12 children (mean age 11.5 years) after a standardised use of various xylitol products: (A) chewing gums (1.3 g xylitol), (B) sucking tablets (0.8 g xylitol), (C) candy tablets (1.1 g xylitol), (D) toothpaste (0.1 g xylitol), (E) rinse (1.0 g xylitol), and (F) a non-xylitol paraffin. Unstimulated saliva was sampled 1, 3, 8, 16 and 30 min after use. The concentration in dental plaque was determined after mouthrinses with contrasting amounts of xylitol (LX = 2.0 g, HX = 6.0 g, and control) and supragingival plaque was collected and pooled after 5, 15 and 30 min. The mean xylitol concentration in saliva at baseline was approximately 0.1 mg/ml. All xylitol-containing products resulted in significantly increased levels (p < 0.05) immediately after intake and remained elevated for 8-16 min in the different groups. The highest mean value in saliva was obtained immediately after use of chewing gums (33.7 +/- 16.4 mg/ml) and the lowest was demonstrated after using toothpaste (8.2 +/- 4.9 mg/ml). No significant differences were demonstrated between chewing gums (A), sucking tablets (B), candy (C) and rinses (E). In dental plaque, the mean values were 8.6 +/- 5.4 and 5.1 +/- 4.0 mg/ml 5 min after HX and LX rinses. Concerning the higher concentration, the values remained significantly elevated (p < 0.05) during the entire 30-min follow-up. In conclusion, commonly advocated xylitol-containing products gave elevated concentrations of xylitol in unstimulated whole saliva and dental plaque for at least 8 min after intake.

Fluoride concentration in saliva after consumption of a dinner meal prepared with fluoridated salt.

The aim was to determine the fluoride concentration in saliva after intake of a dinner meal prepared with fluoridated salt. The investigation had a randomized cross-over design, and 10 healthy adolescents with natural fluoride content (1.06 ppm) in their drinking water participated after informed consent. After a run-in week, the subjects were served a standardized dinner of spaghetti with minced meat sauce prepared with either fluoridated salt (test arm) or non-fluoridated salt (control arm). The fluoride concentration of the test salt was 250 ppm. Samples of stimulated whole saliva was collected at baseline, directly after eating (0 min) and then after 10, 30 and 180 min. After a 1-week wash-out period, the experimental procedure was repeated with the opposite salt. Fluoride concentration in saliva was measured with a fluoride-specific electrode and the post-ingestion levels were compared with baseline using repeated-measures ANOVA. The mean baseline concentrations were 10.9 and 8.0 microg/l in the test and control arms, respectively. Immediately after the intake, the mean fluoride values increased significantly to 81.6 microg/l in the test arm and to 31.5 microg/l in the control arm (p<0.05). The fluoride levels remained elevated (p<0.05) for 30 min after ingestion of the test meal but not following the control meal. In conclusion, consumption of a dinner meal prepared with fluoridated salt increased the salivary fluoride levels for about 30 min.

Fluoride levels in saliva and dental plaque after consumption of snacks prepared with fluoridated salt.

AIM: To determine the concentration of fluoride in saliva and supragingival dental plaque at designated time intervals after consumption of snacks prepared with a standardised amount of fluoridated salt. STUDY DESIGN: The investigation had a single blind prospective crossover design. METHODS: A group of 11 healthy young adults volunteered to participate after verbal and written information and consent. After a 1-week fluoride depletion period, the subjects consumed popcorn prepared with either fluoridated (250 mg/kg) or non-fluoridated salt during 30 minutes. Unstimulated whole saliva and samples of supragingival plaque were collected before consumption (baseline) and at 30, 60 and 120 minutes after the intake. Fluoride concentration was determined with a fluoride-specific electrode and the post-ingestion levels were compared with baseline by ANOVA. RESULTS: In saliva, the mean fluoride concentrations at baseline ranged from 0.021 to 0.027 mg/L and after the 30 minutes consumption of fluoride prepared snacks a 15-fold increase (p<0.001) was found. The same pattern was disclosed in the plaque samples. In both saliva and plaque, the fluoride levels remained significantly elevated after 2 hours (p<0.001 and p<0.05, respectively). CONCLUSION: Consumption of snacks prepared with fluoridated table salt resulted in significantly increased fluoride levels in saliva and supragingival plaque for a period of at least two hours.

The Saccharomyces cerevisiae SOP1 and SOP2 genes, which act in cation homeostasis, can be functionally substituted by the Drosophila lethal(2)giant larvae tumor suppressor gene.

By complementation of a salt-sensitive mutant of Saccharomyces cerevisiae, we cloned the SOP1 gene, encoding a 114.5-kDa protein of 1033 amino acids. Cells deleted for SOP1 exhibited sensitivity to sodium stress, but showed no sensitivity to general osmotic stress. Following exposure of sop1Delta cells to NaCl stress, the intracellular Na+ level and the Na+/K+ ratio rose to values significantly higher than in wild type cells. Deletion of SOP2, encoding a protein sharing 54% amino acid identity with Sop1p, produced only slight Na+ sensitivity. Cells carrying a sop1Deltasop2Delta double deletion became, however, hypersensitive to Na+ and exhibited increased sensitivity also to Li+ and K+, suggesting involvement of both SOP1 and SOP2 in cation homeostasis. The predicted amino acid sequences of Sop1p and Sop2p show significant homologies with the cytoskeletal-associated protein encoded by the Drosophila lethal(2)giant larvae tumor suppressor gene. Immunolocalization of Sop1p revealed a cytoplasmic distribution and cell fractionation studies showed that a significant fraction of Sop1p was recovered in a sedimentable fraction of the cytosolic material. Expression of a Drosophila l(2)gl cDNA in the sop1Deltasop2Delta strain partially restored the Na+ tolerance of the cells, indicating a functional relationship between the Sop proteins and the tumor suppressor protein, and a novel function in cell homeostasis for this family of proteins extending from yeast to human.

Purification and characterisation of a plasminogen-binding protein from Haemophilus influenzae. Sequence determination reveals identity with aspartase.

Plasminogen binding proteins have been described both for Gram positive and Gram negative bacteria. In the present work we describe the purification and characterization of a plasminogen binding protein from Haemophilus influenzae (strain HI-23459). Bacteria were sonicated in order to solubilize plasminogen-binding proteins. The supernatant was subjected to affinity chromatography on plasminogen kringle-4 fragment bound to Sepharose 4B and subsequently processed by ion-exchange chromatography on DEAE-Sepharose CL-6B. Characterization of the protein by SDS-PAGE displayed a single band with a molecular mass of about 55,000, both prior to and after reduction. The purified protein stimulates tPA (tissue plasminogen activator) catalysed plasminogen activation by a factor of approximately 300, mainly due to a decrease in K(m). Antibodies were raised in rabbits and used in quantitative and qualitative analysis. However, using a FITC-conjugate we failed to demonstrate the presence of the purified protein on the surface of intact bacteria. The corresponding gene was isolated from a lambda EMBL3 phage library prepared from chromosomal DNA from the same H. influenzae strain, using an oligonucleotide probe based on the NH2-terminal amino acid sequence. An open reading frame corresponding to 472 amino acid was found. The amino acid sequence of the translated gene demonstrates 97% identity with the recently published sequence from aspartate ammonia lyase (aspartase) from H. influenzae. Enzymatic analysis of the purified protein revealed a high aspartase activity.