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The aims of this study were to describe and validate an in vitro multispecies microbial biofilm model for caries development by evaluating the effects of varying medium concentration of sucrose (0.5 and 1.0%) and fluoride (0.4, 0.8 and 1.0 ppm F) in study 1, and calcium (1.0 and 2.0 mM Ca) in study 2. Defined-multispecies biofilms, formed by Lactobacillus casei, Streptococcus mutans, S. salivarius and S. sanguinis, were grown on the surface of salivary-pellicle-coated enamel slabs, with known baseline surface hardness; growth medium was changed daily. Counts of viable cells on biofilms and the percentage of surface microhardness change (%SMC), lesion depth (LD) and integrated mineral loss (IML) on enamel slabs were assessed after 4 days of biofilm formation under the tested conditions. Counts of viable cells on biofilms were significantly affected by sucrose, fluoride and calcium concentrations (p < 0.05). There was a decrease in %SMC in response to increased fluoride and calcium concentrations (p < 0.001). Lower IML (p < 0.001) and LD (p < 0.05) were found in the presence of 0.8 and 1.0 ppm F. A negative correlation was found between the response variables (%SMC, LD and IML) and fluoride and calcium concentrations. The results suggest that the microbial caries model developed was able to show distinct levels of caries inhibition in response to fluoride and calcium concentrations, corroborating clinical observations. An effect of sucrose concentration on caries development was found only in the presence of the lowest fluoride concentration., (Copyright © 2013 S. Karger AG, Basel.)

Current understanding of dental caries considers this disease a demineralization of the tooth tissues due to the acid produced by sugar-fermenting microorganisms. Thus, caries is considered a diet- and pH-dependent process. We present here the first metagenomic analysis of the bacterial communities present at different stages of caries development, with the aim of determining whether the bacterial composition and biochemical profile are specific to the tissue affected. The data show that microbial composition at the initial, enamel-affecting stage of caries is significantly different from that found at subsequent stages, as well as from dental plaque of sound tooth surfaces. Although the relative proportion of Streptococcus mutans increased from 0.12% in dental plaque to 0.72% in enamel caries, Streptococcus mitis and Streptococcus sanguinis were the dominant streptococci in these lesions. The functional profile of caries-associated bacterial communities indicates that genes involved in acid stress tolerance and dietary sugar fermentation are overrepresented only at the initial stage (enamel caries), whereas other genes coding for osmotic stress tolerance as well as collagenases and other proteases enabling dentin degradation are significantly overrepresented in dentin cavities. The results support a scenario in which pH and diet are determinants of the disease during the degradation of enamel, but in dentin caries lesions not only acidogenic but also proteolytic bacteria are involved. We propose that caries disease is a process of varying etiology, in which acid-producing bacteria are the vehicle to penetrate enamel and allow dentin degrading microorganisms to expand the cavity., (© 2013 S. Karger AG, Basel.)

Background: Establishing animal models with metabolic disorders similar to human metabolic syndrome (MS) is important. In terms of eliciting a full array of MS, we have previously shown that Wistar rats are more responsive to sucrose water drinking than are C57BL/6J mice. This study was aimed at investigating the underlying molecular mechanism of sucrose water-induced MS in Wistar rats., Methods: Male Wistar rats were divided into 2 groups (n = 8 for each group) which were given plain water (C group) or 30% sucrose water (SW group) to drink ad libitum. After 20 weeks, the transcriptional levels and protein translocation of hepatic sterol regulatory element-binding protein-1c (SREBP-1c) and carbohydrate response element-binding protein (ChREBP) as well as the protein levels of protein tyrosine phosphatase-1B (PTP-1B) in insulin-responsive tissues (liver, muscle, and adipose tissue) were measured., Results: The sucrose water regimen successfully elicited visceral obesity, hypertriglyceridemia, insulin resistance, and high blood pressure. The upregulation of de novo lipogenesis in the liver of the sucrose water-treated rats was demonstrated by an increased activity of enzymes, mRNA levels of lipogenic proteins, and nuclear levels of SREBP-1c and ChREBP. Moreover, in the sucrose water-treated rats, protein levels of PTP-1B were significantly increased in liver and skeletal muscle but decreased in adipose tissue., Conclusion: The susceptibility of Wistar rats to sucrose water-induced MS is associated with the transactivation of SREBP-1c and ChREBP in the liver, and PTP-1B is involved in the upregulation of de novo lipogenesis in the liver and the pathology of systemic insulin resistance in rats with MS chronically induced by drinking sucrose water., (Copyright © 2010 S. Karger AG, Basel.)

Background/aims: Isomaltulose (alpha-D-glucosylpyranosyl-1,6-D-fructofuranose) is a natural disaccharide used in human nutrition. It is structurally related to sucrose, but more slowly hydrolyzed and absorbed. Because this sugar's metabolic effects are poorly characterized, we compared the effects of chronic ad libitum access to high-isomaltulose and high-sucrose diets on glucose metabolism in rats., Methods: Adult male rats were offered 62% isomaltulose, sucrose or starch diets ad libitum for 26 (trial 1) or 56 (trial 2) days. After 2- to 3-week adaptation, plasma glucose, fructose and insulin were measured after test meals of the adaptation diet., Results: The main finding was that both plasma glucose and plasma insulin concentrations were transiently but markedly increased after sucrose test meals compared to isomaltulose or starch meals. These differences were not associated with consistent differences in food intake, body weight gain or adiposity., Conclusions: Chronic isomaltulose feeding has beneficial effects on postprandial glucose metabolism in comparison to sucrose feeding in rats, although the effects are modest. Further work is warranted to determine whether substitution of isomaltulose for sucrose or other sweet carbohydrates might be therapeutically useful in patients with, or at risk for, insulin resistance or type 2 diabetes mellitus.

Background/aims: The L/N-type calcium channel blocker (CCB) cilnidipine has been demonstrated to suppress progressive renal disease in a variety of experimental models, but the characteristic effects of N-type calcium channel blocking action on renal injury have not been examined in detail. Therefore, we investigated the beneficial effects of cilnidipine on renal injury in Dahl salt-sensitive (Dahl S) rats fed a high-sucrose diet (HSD), which mimics metabolic syndrome, and compared them with the effects of an L-type CCB, amlodipine., Methods: Male Dahl S rats were divided into groups with similar blood pressure at 8 weeks of age and fed an HSD. They received vehicle, cilnidipine or amlodipine for 27 weeks. At 35 weeks of age, urine and blood samples were collected for physiological analysis, and the kidneys were removed for histopathological evaluation., Results: Cilnidipine reduced albuminuria, glomerular hypertrophy, glomerular expression of ICAM-1, ED-1-positive cell infiltration and interstitial fibrosis compared with vehicle-treated rats. In contrast, amlodipine had no effect on these parameters. Urinary norepinephrine excretion, renal expression of renin mRNA and renal tissue levels of angiotensin II were increased only in the amlodipine-treated group., Conclusion: Cilnidipine provided superior protection against renal damage compared with amlodipine in Dahl S rats given an HSD. The different effects between these two drugs may be partly explained by their different actions on the renal sympathetic nerve activity and the renin-angiotensin system through the N-type calcium channel blocking action.

Unlabelled: Oral clearance and acid production were observed in 30 volunteers following the ingestion of sharp cheddar cheese (CC) and in 9 volunteers following the ingestion of milk chocolate (MC) and low-fat yogurt, and then when MC was eaten immediately after CC (CC/MC) and when CC was eaten following MC (MC/CC). After each test food or food combination had been ingested the volunteers were monitored at five different tooth sites. At each site, using an absorbent paper point, 4 oral fluid samples were collected at 30 min intervals. The five paper points from each sampling occasion were pooled, extracted with 1 ml of water and analyzed qualitatively and quantitatively for both carbohydrates and organic acids using HPLC. Data obtained for each food was averaged and subjected to statistical evaluation. With the CC, glucose clearance was prolonged, due to intermediate lactose degradation into galactose and glucose. The quantity of lactic acid produced during the four intervals was monitored for each of the test foods and their combinations., Results: CC, MC, CC/MC, MC/CC 30 min after ingestion: 1.64, 3.47, 4.68, 2.97; after 60 min: 1.30, 1.24, 1.28, 1.43; after 90 min: 1.58, 1.02, 0.76, 0.43; after 120 min: 1.27, 0.90, 0.70, 0.42 mmol lactic acid/l oral fluid, respectively. The average total amount of lactic acid obtained during the two hour test period was (highest) CC/MC > MC > CC > MC/CC (lowest). The lowest amount of intra-oral lactic acid was observed (Student's t test, p < 0.05) when cheddar cheese (CC) was eaten immediately after the milk chocolate (MC)., Clinical Significance: Consumption of cheese (cheddar), immediately after a sweet meal, significantly reduces (approx. 30%) the amount of lactic acid produced in the oral cavity, when compared to the amount of acid obtained from the sweet food alone., (Copyright 2001 S. Karger AG, Basel)

Background/aims: In order to determine whether the response to a sucrose-rich diet (SRD) during pregnancy in the rat varies depending on the strain, the responsiveness to a SRD (63 g sucrose/100 g) during pregnancy in Wistar and Sprague-Dawley rats was studied., Methods: One group of rats of each strain was fed the SRD, whereas another group received the same diet except that sucrose was replaced by an equal amount of cornstarch. Half of the rats were mated, and all animals were studied 20 days later., Results: Initial body weight did not differ among groups, but final body weight of pregnant Wistar rats was lower than in Sprague-Dawley, and this difference corresponded to a decrease in fetal body weight in the former. Feeding a SRD did not modify pregnancy outcome in either rat strain. Plasma triglycerides increased with a SRD, although this effect was milder in Wistar pregnant rats than in the other groups. Adipose tissue lipoprotein lipase activity was lower in pregnant than in virgin rats, but no differences were found as result of either diet or rat strain. Liver triglyceride concentration increased in virgin rats fed SRD, the effect being greater in Sprague-Dawley than in Wistar rats., Conclusion: Differences in the response to a SRD in pregnant and virgin rats do not depend on the strain of rats used., (Copyright 2001 S. Karger AG, Basel)

Unlabelled: Oral lactic acid production was studied on 11 healthy dental student volunteers (5 males and 6 females) during clearance of five solutions containing 5, 10, 15, 20 and 30% of sucrose. Oral fluid samples were collected at times zero, immediately before taking the sucrose solutions ('baseline'), and 15 min following intake of the solutions. The samples were analyzed qualitatively and quantitatively for the presence of lactic acid and remaining sugars using high-performance liquid chromatography (HPLC). Results indicate that the amount of lactic acid in oral fluid significantly increased with increasing sucrose concentrations, up to 15% sucrose. With the higher sucrose concentrations (20 and 30%) equivalent or lower amounts of lactic acid were obtained. The threshold level for maximum lactic acid production was found to be between 10 and 15% sucrose. Statistical analysis of the data (Student's t test) indicated a significant difference in lactic acid production between the 5 and 10% sucrose solutions versus the 15, 20 and 30% sucrose solutions tested (p < 0.05)., Clinical Significance: Ingestion of solutions with higher sucrose concentrations (>15%) produced similar amounts or less of lactic acid during oral clearance than solutions containing lower sucrose concentrations (<15%)., (Copyright 2000 S. Karger AG, Basel)

The presence of carbohydrates and organic acids was monitored in the oral cavity over a 3-hour period following the ingestion of six foods containing cooked starch (popcorn, potato chips, corn flakes, bread stick, hard pretzel and wheat cracker) and compared to a food containing sugar (chocolate-covered candy bar). Oral fluid samples were collected at 30-min intervals from five different tooth sites from 7 volunteers using absorbent paper points. Samples were analyzed for carbohydrates and organic acids using high-performance liquid chromatography. Analytical data for each food were pooled and compared to the results of the sugar food. The amount of lactic acid produced 30 min after ingestion was highest with the potato chips and lowest with the corn flakes. Potato starch contributed more readily to oral lactic acid production than wheat or corn starch. A direct linear relationship existed between lactic acid production and the presence of oral glucose produced from starch, which occurred via the metabolites maltotriose and maltose. Oral clearance of foods containing cooked starch proceeded significantly slower than that of the sugar food, thus contributing to a prolonged period of lactic acid production.