Effect on oral pH changes and taste perception in 10-14-year-old children, after calcium fortification of a fruit juice.

AIM: The aim was to determine the effect of calcium fortification of a commercially available mixed-fruit juice on oral pH changes and taste perception in a group of 10 to 14 year-old Indian children. METHOD: A controlled, blinded, non-randomised clinical trial was adopted, consisting of a sample of 100 healthy children (DMFT <3; age 10-14 years), who were exposed to three test juices one by one [Group A: original fruit juice (control group); Group B: calcium-fortified fruit juice and Group C: calcium + vitamin D fortified fruit juice]. Oral pH, collection of saliva and plaque sampling was undertaken, before and after the juice exposure by each subject at 0, 1, 5, 15, 30 and 45 min. The respective pH was measured with a digital pH meter. For taste perception, a scoring system was used after exposure of the juices to the subjects in a blind manner. The statistical evaluation was done using one-way ANOVA for salivary and plaque pH and Kruskal-Wallis test for buffer capacity and taste perception. RESULTS: There was a smaller drop in salivary and plaque pH (p < 0.5) and a significant reduction in perceived taste (p < 0.001) by the subjects after calcium modification of fruit juice. CONCLUSION: The calcium-modified mixed fruit juices was less acidogenic compared with the unfortified juice, and hence will be less cariogenic and erosive towards teeth.

An in-vitro assessment of erosive potential of a calcium-fortified fruit juice.

AIM: To evaluate and compare the in vitro pH, buffer capacity and calcium loss from tooth enamel before and after calcium fortification of a packaged fruit juice. METHODS: An approved brand of packaged mixed fruit juice was selected as a test drink on the basis of a pilot questionnaire. The test drink was fortified with 1,000 mg/l (0.1% w/v) of calcium citrate malate to obtain two test groups: Group 1: original beverage (serving as control) and Group 2: calcium-fortified drink. The pH and buffering capacity for the test drinks were measured before and after calcium fortification; 90 prepared enamel samples were divided and immersed into three test subgroups: (1) buffer solution pH 7 (positive control), (2) original fruit juice (negative control) and (3) calcium-fortified fruit juice for 3 min. Calcium loss from the enamel of immersed teeth was measured as a quantitative estimate of tooth mineral loss. RESULTS: After calcium fortification of the fruit juice the mean pH raised from 3.4 to 4.0 (p = 0.029), the mean buffer capacity decreased from 9.73 to 9.16 (p < 0.001) and the mean calcium loss from enamel specimens decreased from 3.5 to 0.26 mg/dl (p < 0.001). STATISTICS: To compare the change in mean pH and buffering capacity between the subject groups, t test was used, and to compare the calcium loss from enamel specimens, among the three subgroups, ANOVA was used. CONCLUSION: Calcium fortification of packaged fruit juice in vitro, improves its pH and buffering capacity. Consequently, the fortified juice causes significantly less mineral loss from human enamel. Fortifying juice with calcium may exert a significant protective potential against dental erosion particularly due to frequent exposure of acidic drinks.

Correlation of susceptibility of Cryptococcus neoformans to amphotericin B with clinical outcome.

Testing of Cryptococcus neoformans for susceptibility to antifungal drugs by standard microtiter methods has not been shown to correlate with clinical outcomes. This report describes a modified quantitative broth macrodilution susceptibility method showing a correlation with both the patient's quantitative biological response in the cerebrospinal fluid (CSF) and the survival of 85 patients treated with amphotericin B (AMB). The Spearman rank correlation between the quantitative in vitro measure of susceptibility and the quantitative measure of the number of organisms in the patient's CSF was 0.37 (P < 0.01; 95% confidence interval [95% CI], 0.20, 0.60) for the first susceptibility test replicate and 0.46 (P < 0.001; 95% CI, 0.21, 0.62) for the second susceptibility test replicate. The median in vitro estimated response (defined as the fungal burden after AMB treatment) at 1.5 mg/liter AMB for patients alive at day 14 was 5 CFU (95% CI, 3, 8), compared to 57 CFU (95% CI, 4, 832) for those who died before day 14. These exploratory results suggest that patients whose isolates show a quantitative in vitro susceptibility response below 10 CFU/ml were more likely to survive beyond day 14.

Detection of differentially expressed genes in synovial fibroblasts by restriction fragment differential display.

OBJECTIVE: To identify differentially expressed genes in synovial fibroblasts and examine the effect on gene expression of exposure to TNF-alpha and IL-1beta. METHODS: Restriction fragment differential display was used to isolate genes using degenerate primers complementary to the lysophosphatidic acid acyl transferase gene family. Differential gene expression was confirmed by reverse transcription-polymerase chain reaction and immunohistochemistry using a variety of synovial fibroblasts, including cells from patients with osteoarthritis and self-limiting parvovirus arthritis. RESULTS: Irrespective of disease process, synovial fibroblasts constitutively produced higher levels of IL-6 and monocyte chemoattractant protein 1 (MCP-1) (CCL2) than skin fibroblasts. Seven genes were differentially expressed in synovial fibroblasts compared with skin fibroblasts. Of these genes, four [tissue factor pathway inhibitor 2 (TFPI2), growth regulatory oncogene beta (GRObeta), manganese superoxide dismutase (MnSOD) and granulocyte chemotactic protein 2 (GCP-2)] were all found to be constitutively overexpressed in synoviocytes derived from patients with osteoarthritis. These four genes were only weakly expressed in other synovial fibroblasts (rheumatoid and self-limiting parvovirus infection). However, expression in all types of fibroblasts was increased after stimulation with TNF-alpha and IL-1beta. Three other genes (aggrecan, biglycan and caldesmon) were expressed at higher levels in all types of synovial fibroblasts compared with skin fibroblasts even after stimulation with TNF-alpha and IL-1. CONCLUSIONS: Seven genes have been identified with differential expression patterns in terms of disease process (osteoarthritis vs rheumatoid arthritis), state of activation (resting vs cytokine activation) and anatomical location (synovium vs skin). Four of these genes, TFPI2, GRObeta (CXCL2), MnSOD and GCP-2 (CXCL6), were selectively overexpressed in osteoarthritis fibroblasts rather than rheumatoid fibroblasts. While these differences may represent differential behaviour of synovial fibroblasts in in vitro culture, these observations suggest that TFPI2, GRObeta (CXCL2), MnSOD and GCP-2 (CXCL6) may represent new targets for treatments specifically tailored to osteoarthritis.

Amphotericin B colloidal dispersion combined with flucytosine with or without fluconazole for treatment of murine cryptococcal meningitis.

Studies with animals and in vitro studies have demonstrated that flucytosine plus amphotericin B or fluconazole has significantly improved mycologic activity against meningitis caused by Cryptococcus neoformans compared to the activity of amphotericin B or fluconazole used alone. However, few doses have been tested in combination. This study evaluated the antifungal efficacy of amphotericin B colloidal dispersion (ABCD) combined with flucytosine with and without fluconazole in a murine model of cryptococcal meningitis. The following dosages were tested: ABCD at 0 to 12.5 mg/kg of body weight given intravenously 3 days/week, flucytosine at 0 to 110 mg/kg/day, and fluconazole at 0 to 50 mg/kg/day. Meningitis was established in male BALB/c mice by intracerebral injection of C. neoformans. Treatment with flucytosine with or without fluconazole dissolved in the sole source of drinking water was started on day 2; animals were sacrificed at 16 days, and the numbers of fungal colonies in the brain were quantified. A survival rate of 100% was achieved with ABCD plus flucytosine without fluconazole; however, the addition of fluconazole was required to prevent weight loss (P < 0.00001) and to achieve the maximum antifungal effect (P < 0.00001). The only region of dose combinations for which the 99% confidence intervals were less than 100 CFU/g of brain was defined by ABCD at 5.0 to 7.5 mg/kg combined with flucytosine at 20 to 60 mg/kg/day and fluconazole at 30 to 40 mg/kg/day. The triple combination of ABCD plus flucytosine and fluconazole was necessary to achieve the greatest antifungal activity.

Effect of severity of meningitis on fungicidal activity of flucytosine combined with fluconazole in a murine model of cryptococcal meningitis.

We studied the effect of the severity of meningitis on the response to therapy with fluconazole and flucytosine in a murine model of cryptococcal meningitis. Meningitis was established by intracerebral injection of Cryptococcus neoformans. The severity of meningitis was varied by delaying the onset of treatment from 3 to 7 days. Animals were sacrificed after 14 days of treatment, and the numbers of C. neoformans per gram of brain tissue were quantified. The range of effective dose combinations of fluconazole and flucytosine became progressively reduced as the severity of meningitis increased. The magnitude of treatment effect, as measured by the numbers of CFU/gram of brain tissue, was also reduced with increasing severity of meningitis. In this model, as the severity of meningitis increases, higher doses of fluconazole are required to achieve equivalent levels of activity. The combination of fluconazole and flucytosine appears to have the most-potent antifungal effects. This is most readily observed in animals with more-severe meningitis.