The aim of this study was to make cellular-level measurements of the mechanical efficiency of mouse cardiac muscle and to use these measurements to determine (1) the work performed by a cross-bridge in one ATP-splitting cycle and (2) the fraction of the free energy available in metabolic substrates that is transferred by oxidative phosphorylation to free energy in ATP (i.e. mitochondrial thermodynamic efficiency). Experiments were performed using isolated left ventricular mouse papillary muscles (n = 9; studied at 27°C) and the myothermic technique. The production of work and heat was measured during and after 40 contractions at a contraction frequency of 2 Hz. Each contraction consisted of a brief isometric period followed by isovelocity shortening. Work output, heat output and enthalpy output were all independent of shortening velocity. Maximum initial mechanical efficiency (mean ± SEM) was 31.1 ± 1.3% and maximum net mechanical efficiency 16.9 ± 1.5%. It was calculated that the maximum work per cross-bridge cycle was 20 zJ, comparable to values for mouse skeletal muscle, and that mitochondrial thermodynamic efficiency was 72%. Analysis of data in the literature suggests that mitochondrial efficiency of cardiac muscle from other species is also likely to be between 70 and 80%.

Sea buckthorn berries contain many bioactive compounds that have anticancer properties. To investigate whether the anti proliferative effects Could be associated with the presence of certain compounds. a sequential extraction was performed. The extraction started with heptane followed by ethyl acetate, ethanol, and water. A second protocol using ethanol:water (1:1) was also used. The contents of the extracts were determined and their effects on cell proliferation were investigated in both Caco-2 and Hep G2 cells. The ethyl acetate fraction was exclusively found to contain high levels of ursolic acid, together with low amounts of phenolics. The ethanol:water extracts contained high levels of phenolic compounds and proanthyocyanidin, but little ursolic acid. When the antiproliferative effects were examined, the strongest inhibitory effect was found in the ethyl acetate extract for the Caco-2 cells and in the ethanol:water extract for the Hep G2 cells. The antiproliferative effects were in both cases dose-dependent and were in the case of the ethyl acetate extract associated with an increase in apoptosis. The results obtained show that the choice of extraction solvent is of considerable importance and that ursolic acid might be more important than the polyphenols in inhibiting the cancer cell proliferation. (C) 2009 Elsevier Ltd. All rights reserved.

Objective.

The aim of this double-blind randomized placebo-controlled trial was to evaluate the effects of probiotic supplements in adjunct to conventional management of peri-implant mucositis.

Materials and methods.

Forty-nine adult patients with peri-implant mucositis were consecutively recruited after informed consent. After initial mechanical debridement and oral hygiene instructions, the patients received a topical oil application (active or placebo) followed by twice-daily intake of lozenges (active or placebo) for 3 months. The active products contained a mix of two strains of Lactobacillus reuteri. Patients were clinically monitored and sampled at baseline and after 1, 2, 4, 12 and 26 weeks. The clinical end-points were pocket-probing depth (PPD), plaque index (PI) and bleeding on probing (BOP). In addition, the subgingival microbiota was processed with checkerboard DNA-DNA hybridization and samples of gingival crevicular fluid (GCF) were analyzed for selected cytokines with the aid of multiplex immunoassays.

Results.

After 4 and 12 weeks, all clinical parameters were improved in both the test and the placebo group. PPD and BOP were significantly reduced compared with baseline (p < 0.05), but no significant differences were displayed between the groups. The clinical improvements persisted 3 months after the intervention. No major alterations of the subgingival microflora were disclosed and the levels of inflammatory mediators in GCF did not differ between the groups.

Conclusions.

Mechanical debridement and oral hygiene reinforcement resulted in clinical improvement of peri-implant mucositis and a reduction in cytokine levels. Probiotic supplements did not provide added benefit to placebo.

Harpagophytum, Devil's Claw, is a genus of tuberiferous xerophytic plants native to southern Africa. Some of the taxa are appreciated for their medicinal effects and have been traditionally used to relieve symptoms of inflammation. The objectives of this pilot study were to investigate the antioxidant capacity and the content of total phenols, verbascoside, isoverbascoside, and selected iridoids, as well as to investigate the capacity of various Harpagophytum taxa in suppressing respiratory burst in terms of reactive oxygen species produced by human neutrophils challenged with phorbol myristate acetate (PMA), opsonised Staphylococcus aureus, and Fusobacterium nucleatum. Harpagophytum plants were classified into different taxa according to morphology, and DNA analysis was used to confirm the classification. A putative new variety of H. procumbens showed the highest degree of antioxidative capacity. Using PMA, three Harpagophytum taxa showed anti-inflammatory effects with regard to the PBS control. A putative hybrid between H. procumbens and H. zeyheri in contrast showed proinflammatory effect on the response of neutrophils to F. nucleatum in comparison with treatment with vehicle control. Harpagophytum taxa were biochemically very variable and the response in suppressing respiratory burst differed. Further studies with larger number of subjects are needed to corroborate anti-inflammatory effects of different taxa of Harpagophytum.

AIM: The aim was to assess clinical inflammatory parameters, cytokine levels, and bacterial counts in samples from implant crevicular fluid in cases with untreated peri-implantitis.

MATERIAL AND METHODS: Several bacterial species known to up-regulate pro-inflammatory cytokines have been associated with peri-implantitis. The Luminex magnet bead technology was used to study cytokines in crevicular fluid. The checkerboard DNA-DNA hybridization method was used to study bacterial counts in samples from 41 implants (41 individuals).

RESULTS: Profuse bleeding and suppuration was found in 25/41 (61.0%) and 17/41 (41.5%) of the implants. The reliability of duplicate cytokine processing was high. In the presence of profuse bleeding, higher pg/ml levels of IL-1β (p = 0.016), IL-8 (p = 0.003), TNF-α (p = 0.024), and VEGF (p = 0.004) were found. Higher concentrations of IL-1β were found in the presence of suppuration, and if Escherichia coli (p = 0.001) or Staphylococcus epidermidis (p = 0.05) could be detected.

CONCLUSION: Profuse bleeding and/or suppuration in untreated peri-implantitis can be associated with higher concentrations of IL-1β, IL-8, TNF-α and VEGF in peri-implant crevicular fluid. A higher concentration of IL-1β in peri-implant crevicular fluid was found in samples that were positive for E. coli or S. epidermidis.

AIM: To study whether cytokine levels and bacterial counts in p atients with peri-implantitis reflect clinical treatment outcome following non-surgical management.

MATERIALS AND METHODS: Luminex magnet bead technology and checkerboard DNA-DNA hybridization were used to assess treatment outcome after treatment at the implant with the most severe peri-implantitis in 41 participants.

RESULTS: Study group mean age was 40.3 years (SD ± 9.9). Stable treatment outcome after 6 months (no further bone loss, probing pocket depth decrease ≥0.5 mm, no bleeding/suppuration) was identified in 9 of 41 (22%) participants. Peri-implant crevicular fluid (PICF) levels were also lower for Il-1β (P < 0.01), and with trends of lower cytokine levels in PICF for TNF-α (P = 0.071), PDGFBB (P = 0.071), as well as for VEGF (vascular endothelial growth factor) (P = 0.071), and bacterial counts for Actinomyces israelii, Aggregatibacter actonomycetemcomitans (Y4), Campylobacter gracilis, Echerichia coli, Fusobacterium periodonticum, Leptotrichia buccalis, Parvimonas micra, Staphylococcus haemolyticus, Streptococcus anginosus, and Tannerella forsythia. Increasing levels of IL-1 β and S. aureus (r(2)  = 0.856) were found only at implants with non-stable outcome. A reduction of PICF levels for selected cytokines and bacteria studied had a sensitivity of 0.77, and a specificity of 0.80 against the clinical outcome as gold standard. Data analysis failed to differences in treatments (PerioFlow(®) versus YAG: ER laser) for changes in the expression of cytokines and bacteria studied.

CONCLUSIONS: At 6 months, clinically stable treatment outcome of peri-implantitis is associated lower levels of putative pathogens total bacterial load with ≥30% reduction of IL1-β, L-6, and VEGF levels in PICF.

Berries contain several bioactive compounds that can protect against oxidative stress. In this study we evaluated the protective effect of different sequential extracts (ethyl acetate, ethanol and water) of seven berry species: bilberry (Vaccinium myrtillus), blackcurrant (Ribes nigrum), elderberry (Sambucus nigra), lingonberry (Vaccinium vitis-idaea), rose hips (Rosa sp.), sea buckthorn (Hippohae rhamnoides) and strawberry (Fragaria × ananassa). The protective effect was tested on human erythrocytes and the antioxidant capacity was also evaluated in vitro by the FRAP assay. In the erythrocyte assay all sea buckthorn extracts were superior in antioxidant effect to other berry extracts. The ethyl acetate extract of bilberries, and the ethanol and water extracts of blackcurrants, also protected the erythrocytes from oxidation. In contrast, water extracts of rose hips, bilberries and strawberries had a pro-oxidant effect on erythrocytes. The water extract of rose hips was superior to the other berry extracts in the FRAP assay. Thus, the results of the erythrocyte assay did not correlate with the results of the FRAP assay, but provided additional insights into the potential protective effects of berry extracts against oxidative stress.

Bioactive molecules in berries may be helpful in reducing the risk of oral diseases. The aim of this study was to determine the effect of bilberry consumption on the outcome of a routine dental clinical parameter of inflammation, bleeding on probing (BOP), as well as the impact on selected biomarkers of inflammation, such as cytokines, in gingival crevicular fluid (GCF) in individuals with gingivitis. Study individuals who did not receive standard of care treatment were allocated to either a placebo group or to groups that consumed either 250 or 500 g bilberries daily over seven days. The placebo group consumed an inactive product (starch). A study group, receiving standard of care (debridement only) was also included to provide a reference to standard of care treatment outcome. Cytokine levels were assayed using the Luminex MagPix system. The mean reduction in BOP before and after consumption of test product over 1 week was 41% and 59% in the groups that consumed either 250 or 500 g of bilberries/day respectively, and was 31% in the placebo group, and 58% in the standard of care reference group. The analysis only showed a significant reduction in cytokine levels in the group that consumed 500 g of bilberries/day. A statistically significant reduction was observed for IL-1b (p = 0.025), IL-6 (p = 0.012) and VEGF (p = 0.017) in GCF samples in the group that consumed 500 g of bilberries daily. It appears that berry intake has an ameliorating effect on some markers of gingival inflammation reducing gingivitis to a similar extent compared to standard of care.

There is currently an interest in the possible anti-inflammatory effects of intake of fruits and berries. The aim of this study was to determine whether the twice daily administration of a berry beverage rich in polyphenols had effects on salivary levels of a selected group of pro-inflammatory cytokines for one week in a pre-and post-study design. Levels of selected cytokines were compared in whole saliva and saliva obtained using commercially available collection devices (Salivette (R) Cotton and Salivette (R) Synthetic rolls). Twenty healthy subjects drank 200 mL of a berry beverage consisting of equal parts of bilberries (Vaccinium myrtillus), black currant (Ribes nigrum), lingonberries (Vaccinium vitis-idaea), sea buckthorn (Hippophae rhamnoides) diluted with 50% water. Levels of cytokines, IL-1 beta, IL-8, IL-12 and TNF-alpha were assessed. Levels of cytokines differed between sources of collection but were highest in whole saliva. The use of cotton or synthetic rolls does not seem to be useful as a method for saliva collection and cytokine analysis. There was no significant change in the levels of selected cytokines at baseline and after intake of the berry beverage in whole stimulated saliva. There was a large inter-individual variation in cytokine levels.

Rose hips are popular in health promoting products as the fruits contain high content of bioactive compounds. The aim of this study was to investigate whether health benefits are attributable to ascorbic acid, phenols, or other rose-hip-derived compounds. Freeze-dried powder of rose hips was preextracted with metaphosphoric acid and the sample was then sequentially eluted on a C₁₈ column. The degree of amelioration of oxidative damage was determined in an erythrocyte in vitro bioassay by comparing the effects of a reducing agent on erythrocytes alone or on erythrocytes pretreated with berry extracts. The maximum protection against oxidative stress, 59.4±4.0% (mean ± standard deviation), was achieved when incubating the cells with the first eluted meta-phosphoric extract. Removal of ascorbic acid from this extract increased the protection against oxidative stress to 67.9±1.9%. The protection from the 20% and 100% methanol extracts was 20.8±8.2% and 5.0±3.2%, respectively. Antioxidant uptake was confirmed by measurement of catechin by HPLC-ESI-MS in the 20% methanol extract. The fact that all sequentially eluted extracts studied contributed to protective effects on the erythrocytes indicates that rose hips contain a promising level of clinically relevant antioxidant protection.

AIM: A causative relationship between acute coronary syndrome (ACS) and periodontitis has yet to be defined. The aim of this study was to assess differences in levels of serum cytokines between individuals with or without ACS or periodontal comorbidity.

MATERIAL AND METHODS: In a case-control study, individuals with ACS (78 individuals, 10.3% females) and matching healthy controls (78 individuals, 28.2% females) were included. Medical and dental examinations were performed to diagnose ACS and periodontitis. Serum levels of cytokines were assessed using Luminex technology.

RESULTS: A diagnosis of periodontitis in the ACS and control group was diagnosed in 52.6% and 12.8% of the individuals, respectively. The unadjusted odds-ratio that individuals with ACS also had periodontitis was 7.5 (95% CI: 3.4, 16.8, p<0.001). Independent of periodontal conditions, individuals with ACS had significantly higher serum levels of IL8 (mean: 44.3 and 40.0 pg/ml) and vascular endothelial growth factor (VEGF) (mean: 82.3 and 55.3 pg/ml) than control individuals. A diagnosis of periodontitis made no difference in serum cytokine expressions.

CONCLUSION: Elevated serum levels of VEGF were associated with ACS. Serum cytokine expression in individuals with ACS is unrelated to periodontal conditions. This article is protected by copyright. All rights reserved.

Objective

The aim of the present study was to evaluate in vitro antibacterial activities of blackcurrant and sea buckthorn juices on bacteria associated with gingival inflammation.

Materials and methods

The growth of selected bacteria (Streptococcus mitis, Streptococcus mutans, Streptococcus sanguinis, Streptococcus gordonii, Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa) was studied in vitro on agar plates. The content of phenols in the different extracts was measured with HPLC-ESI-MS.

Results

The spectrometric analysis identified that the highest level of the single phenols studied was found for ferulic acid (113 μg/ml) in blackcurrant juice. Sea buckthorn contained low levels of selected phenols. Total bacterial inhibition for all bacterial species studied was found at 20% berry juice concentration with pH varying between 4.1-5.4.

Conclusions

The present study identified that in vitro bacterial growth on agar plates was inhibited by blackcurrant and sea buckthorn juices and that low juice pH explains bacterial in vitro growth. This may have clinical implications in biofilm development, reducing the risks for both tooth decay and gingivitis.

Aim of study The present study was undertaken to evaluate the effects from different leaf extracts of the traditional medicinal herb Plantago major L. (plantain) on cell proliferation and migration in vitro, as a test for potential wound healing properties. Materials and methods Water and ethanol-based extracts were prepared from Plantago major fresh and dried leaves, and tested in vitro in a scratch assay with oral epithelial cells. Results The scratch assay produced reliable results after 18&#xa0;h. Most of the tested extracts increased the proliferation/migration of the oral epithelial cells compared to the negative control. A concentration of 1.0&#xa0;mg/mL (on dry weight basis) appears to be optimal regardless of type of extract, and among the alternatives, 0.1&#xa0;mg/mL was always better than 10&#xa0;mg/mL. Ethanol-based extracts with a concentration of 10&#xa0;mg/mL had very detrimental effects on cell proliferation/migration. At the other two concentrations, ethanol-based extracts had the most beneficial effect, followed by water extracts of fresh leaves, ethanol plus water extracts of dried leaves and, finally, water extracts of dried leaves. This study suggests that both the water extracts and the more polyphenol-rich ethanol-based extracts of Plantago major leaves have medicinal properties. Further research is, however, needed to determine what compounds are responsible for the wound healing effects.