Evaluation of plant and fungal extracts for their potential antigingivitis and anticaries activity.

The link between diet and health has lead to the promotion of functional foods which can enhance health. In this study, the oral health benefits of a number of food homogenates and high molecular mass and low molecular mass fractions were investigated. A comprehensive range of assays were performed to assess the action of these foods on the development of gingivitis and caries using bacterial species associated with these diseases. Both antigingivitis and anticaries effects were investigated by assays examining the prevention of biofilm formation and coaggregation, disruption of preexisting biofilms, and the foods' antibacterial effects. Assays investigating interactions with gingival epithelial cells and cytokine production were carried out to assess the foods' anti- gingivitis properties. Anti-caries properties such as interactions with hydroxyapatite, disruption of signal transduction, and the inhibition of acid production were investigated. The mushroom and chicory homogenates and low molecular mass fractions show promise as anti-caries and anti-gingivitis agents, and further testing and clinical trials will need to be performed to evaluate their true effectiveness in humans.

Non-HDL cholesterol predicts coronary heart disease in primary prevention: findings from an Italian 40-69 year-old cohort in general practice.

OBJECTS: Non-HDL cholesterol is now recommended as an index of risk associated with combined dyslipidemia, and it has also been found useful in predicting coronary heart disease (CHD) risk in patients with diabetes. We studied the association between known CHD risk factors, enclosed non-HDL cholesterol, and a "high CHD risk condition", i.e. a "5-years CHD risk >15%" in general practice. METHODS: We studied 4,085 40-69 year-old diabetic (no. 489) and non-diabetic (no. 3,596) individuals from an opportunistic cohort. Cross-sectional descriptive statistics, and age- and gender-adjusted multiple logistic exponential betas have been calculated. RESULTS: About 12% of the participants had diabetes. Age- and gender-adjusted comparison showed that all the study variables were significantly worse in diabetic vs. non-diabetic individuals (except cigarette smoking, total blood cholesterol and the ratio of total to HDL cholesterol). They had a mean "5-year CHD-risk" significantly higher than non-diabetic individuals (18.8+/-11.9% vs 7.5+/-6.9%, P<0.01), and a four-fold prevalence of "5-years CHD risk >15%" (55.4% vs 11.1%, P<0.01). As to diabetic individuals, the study variables associated to a "high CHD risk condition" were cigarette smoking, systolic blood pressure, and non-HDL blood cholesterol levels. As to non-diabetic individuals cigarette smoking, systolic blood pressure, and HDL (inversely) and non-HDL blood cholesterol levels were associated to a "high CHD risk condition". CONCLUSIONS: Non-HDL cholesterol--and cigarette smoking and systolic blood pressure--strongly predicted a "high CHD risk condition" both in diabetic and non-diabetic individuals.

In vitro and ex vivo anti- and prooxidant components of Cichorium intybus.

The water soluble antioxidant properties of Cichorium intybus var. Silvestre, whose production zone is around Chioggia, Italy, were investigated. Vegetable juices were obtained by centrifugation, and (1) filtration at 2 degrees C; (2) filtration at 25 degrees C, and stored for 3 h; (3) boiled for 30 min at 102 degrees C, and then analysed. The antioxidant properties were evaluated in vitro as antioxidant activity (AA) (model system beta-carotene-linoleic acid) and ex vivo as protective activity (PA) against rat liver cell microsome lipid peroxidation measured as 2-thiobarbituric acid-reactive substances (TBA-RS) generated by peroxide degradation. All the vegetable juices showed high but very variable AA (> 83%) and PA (> 64%). After dialysis and analysis of fractions it was shown that the vegetable contained both biological antioxidant and prooxidant compounds. The prooxidants had MW < 3000, conversely the very active antioxidants (PA = 100%) had MW > 15,000. Electrophoretic analysis revealed that the most active fraction was a complex mixture of brown components at MW > 300,000.

In vitro antioxidant and ex vivo protective activities of green and roasted coffee.

The antioxidant properties of green and roasted coffee, in relation to species (Coffea arabica and Coffea robusta) and degree of roasting (light, medium, dark), were investigated. These properties were evaluated by determining the reducing substances (RS) of coffee and its antioxidant activity (AA) in vitro (model system beta-carotene-linoleic acid) and ex vivo as protective activity (PA) against rat liver cell microsome lipid peroxidation measured as TBA-reacting substances. RS of C. robustasamples were found to be significantly higher when compared to those of C. arabica samples (p < 0.001). AA for green coffee samples were slightly higher than for the corresponding roasted samples while PA was significantly lower in green coffee compared to that of all roasted samples (p < 0.001). Extraction with three different organic solvents (ethyl acetate, ethyl ether, and dichloromethane) showed that the most protective compounds are extracted from acidified dark roasted coffee solutions with ethyl acetate. The analysis of acidic extract by gel filtration chromatography (GFC) gave five fractions. Higher molecular mass fractions were found to possess antioxidant activity while the lower molecular mass fractions showed protective activity. The small amounts of these acidic, low molecular mass protective fractions isolated indicate that they contain very strong protective agents.

Isolation of an antibacterial component from roasted coffee.

A coffee beverage obtained from instant dark coffee that had been previously shown to possess high antibacterial activity, was acidified (pH 2) and extracted with ethyl acetate. After alkalinization (pH 12) the aqueous phase was re-extracted with the organic solvent. The acidic and basic extracts were evaporated to dryness and the aqueous phase freeze-dried. Residues were dissolved in sterile water and assayed for antibacterial activity against two reference bacteria (Staphylococcus aureus ATCC 25923 and Streptococcus mutans 9102). The acidic extract was found to be highly active and was separated by gel permeation chromatography (GPC) into five fractions. Fractions GPC4 and GPC5 were found to possess antibacterial activity: most of the activity was evident in fraction GPC5. These fractions were separated by RP-HPLC using a gradient elution with methanol water as mobile phase. Both GPC fractions gave an active subfraction with methanol-water (70:30, v/v). The experimental conditions used to separate the antibacterial compound that originates during the roasting process, indicate that it possesses low molecular mass (probably no more than 200 Da), weak acidic properties and an lambda(max) at 205 nm. The very small amount of this compound isolated from roasted coffee, indicates that it may be a very strong antibacterial agent.

Increased cAMP levels in human hair follicles due to local treatment with trichoriboside.

Local therapy with trichoriboside and trichosaccharide, which have been found to be beneficial for scalp hair maintenance in adult males affected by androgenic alopecia, was found to increase cAMP levels in human scalp hair follicles. The increase was significant in men affected by androgenic alopecia, whereas it was not significant in unaffected control men. Trichoriboside showed a greater activity than trichosaccharide, and such activity was accompanied by a significant concomitant reduction of ATP in the hair.

Pharmacokinetics of dipyridamole-beta-cyclodextrin complex in healthy volunteers after single and multiple doses.

Dipyridamole is a well known anti-aggregating agent characterized by poor water solubility as well as scant and variable bioavailability. Recently, the compound was complexed with beta-cyclodextrin forming a molecular encapsulation resulting in better oral absorption and stronger biological activities in animals. In the present study, a randomized double blind cross-over comparison between dipyridamole-beta-cyclodextrin complex (dip-beta-CD) and dipyridamole was performed in 12 healthy subjects after single (75mg) and multiple oral treatments (75mg TID). Dip-beta-CD showed better bioavailability and less interindividual variability than dipyridamole either after single or multiple doses. In particular, dip-beta-CD had a greater AUC and Cmax, and a smaller Tmax even at the steady state. In addition, 100% of the subjects receiving a single dose of dip-beta-CD, as compared to 66.7% of those treated with dipyridamole, had plasma levels superior to 1 microgram/ml (which is the supposed anti-aggregating threshold level). In contrast, 0 and 33.03% of the subjects showed plasma levels superior to 2.5 micrograms/ml (which might cause the appearance of side-effects) on the 7th day of the multiple treatment with dip-beta-CD and dipyridamole, respectively. In fact, the subjects presenting higher levels after uncomplexed dipyridamole also complained of headache and/or dizziness on occasion. No adverse side effects were reported for dip-beta-CD.

Erdosteine protection from cigarette smoke-induced loss of alpha 1-antitrypsin activity in rat lungs.

Acute inhalation of cigarette smoke induced alpha 1-antitrypsin inactivation in rat lungs. The elastase inhibitory capacity of the protein could be prevented by oral treatment with erdosteine (a mucoregulator endowed with free radical scavenging properties). A partial dose-dependent protection was observed with 500 and 1,000 mg/kg. The mechanism of its action might be related either to the inhibition of alpha 1-antitrypsin oxidation manifested by two SH groups liberated in vivo or to the anti-inflammatory and mucoregulator properties present in the substance.

Effect of storage on mutagenicity, absorbance and content of furfural in the ribose-lysine maillard system.

The stability with time of browned mixtures characterized by different water activities (aw 0.98, 0.84 and 0.60) and heating temperatures (100, 120, 140 and 160 degrees C) was analysed using the ribose-lysine model system. The results obtained demonstrated the occurrence, during storage, of changes both in the composition of the browned mixtures and in their mutagenic properties, as detected with the Ames test. Only the browned mixtures obtained at 100 degrees C showed a progressive increase in mutagenicity during storage, matched by an increase in ultraviolet and visible light absorbance and of furfural content. In the browning mixtures characterized by a more advanced stage of the Maillard reaction, in which mutagenic activity had initially been found to be undetectable, such activity became evident during storage.

In vitro protection by erdosteine against oxidative inactivation of alpha-1-antitrypsin by cigarette smoke.

Direct exposure in vitro of the protein alpha 1-antitrypsin (alpha 1-AT; human neutrophil elastase inhibitor, alpha 1-proteinase inhibitor) to gas phase cigarette smoke causes a loss of elastase-inhibitory capacity (EIC). This effect appears to be related to the formation of reactive oxygen species in the smoke that inactivate alpha 1-AT by oxidizing the methionine terminal amino acid. Reducing agents such as glutathione and ascorbic acid prevent this inactivation. In the present investigation erdosteine, a novel thiol derivative, which contains two blocked SH groups with potential reducing properties, was tested in vitro for its capacity to protect human alpha 1-AT. For the purpose, the compound, previously hydrolyzed with bicarbonate-carbonate buffer or with microsomal enzymes was put in contact with alpha 1-AT and exposed to gas phase cigarette smoke. The EIC of alpha 1-AT was then measured by incubating the samples with leukocyte elastase and, subsequently, by titrating the residual elastolytic activity against a synthetic substrate. Under these conditions erdosteine effectively protected alpha 1-AT against the smoke injury and, after alkaline hydrolysis, it appeared to be as active as glutathione and ascorbic acid (EC50 being respectively 6.4, 7.2 and 6.2 mM). This evidence suggests that the erdosteine SH groups, which can become free, may have an important role in the mechanism of action, by blocking highly reactive oxygen-free radicals. Erdosteine may have a therapeutic application in preventing oxidative lung damage induced by cigarette smoke.

Influence of water activity and reaction temperature of ribose-lysine and glucose-lysine Maillard systems on mutagenicity, absorbance and content of furfurals.

The effect of water activity (aw 0.98, 0.84 and 0.60) and reaction temperature (100, 120, 140 and 160 degrees C) on the mutagenic activity of the Maillard reaction products in heated ribose-lysine and glucose-lysine model systems, was investigated. In the ribose-lysine system, heated at 100 degrees C, the mutagenic activity of the mixture increased as the water activity was lowered. On the contrary, no dependence between mutagenic activity and water activity was observed in the glucose-lysine system. At higher temperatures, in both systems, the presence in the browned mixtures of an antibacterial activity interfering with the bacterial mutagenicity assay was observed. Under all the conditions tested, the ribose-lysine system turned out to be the most reactive by producing higher levels of mutagens. Furthermore, in this system, the antimicrobial interference was more easily detectable. In the model systems used, the browning reaction mixtures were analysed for their absorption spectrum between 200-460 nm, and for the accumulation of furfurals. The results obtained showed that, at temperatures between 120 and 140 degrees C there is a correlation among reaction temperature, absorbance at 420 and around 280 nm, mutagenic activity of the mixture and the level of furfurals. Changes in the levels of furfurals can be related to changes in mutagenicity of the browned mixtures.

Human pharmacokinetics of erythromycin propionate-N-acetylcysteinate: comparative evaluation with erythromycin stearate and N-acetylcysteine.

The pharmacokinetic pattern of erythromycin propionate-N-acetylcysteinate (EPAC) (erythromycin stinoprate I.N.N.), a new derivative, was studied on 12 healthy volunteers after single and multiple oral treatments. Microbiological and/or HPLC analytical methods were used to titer either erythromycin as base, propionate and total or N-acetylcysteine (NAC). In the acute experiment, a comparative evaluation was performed with erythromycin stearate (ES) and with N-acetylcysteine, according to a randomized-multi-crossover design. EPAC showed a better bioavailability than ES with longer-lasting serum levels of active antibiotic. NAC concentrations in the serum after EPAC were practically identical to those found after an oral administration of NAC alone. The multiple treatment study, performed in the same 12 volunteers with only EPAC, indicated that the pharmacokinetic pattern is somewhat different from that observed after a single dose, since higher concentrations were present at the steady state conditions.

Influence of erdosteine, a mucolytic agent, on amoxycillin penetration into sputum in patients with an infective exacerbation of chronic bronchitis.

Twenty four patients with acute infective exacerbations of chronic bronchitis received amoxycillin alone or in combination with erdosteine (a mucolytic agent) for a week in a double blind, placebo controlled study. Clinical assessment scores, body temperature, serum and sputum amoxycillin concentrations, and sputum culture results were recorded in each group. Erdosteine significantly increased antibiotic concentrations in sputum but not in serum. The combined treatment also caused a more rapid decrease in sputum viscosity and in body temperature and faster sterilisation of the sputum. These results show that erdosteine increases amoxycillin concentration in sputum in patients with acute exacerbations of chronic bronchitis. This effect may be due to a reduction in the viscosity of the bronchial secretions produced by erdosteine.

Serum, sputum and bronchial concentrations of erythromycin in chronic bronchitis after single and multiple treatments with either propionate-N-acetylcysteinate or stearate erythromycin.

Serum and bronchial concentrations of erythromycin were determined in 30 chronic bronchitic patients during an exacerbation phase of bacterial infections. The levels were measured after single and multiple oral treatments with erythromycin-propionate-N-acetylcysteinate (EPAC) or erythromycin stearate (ES) in a double-blind design. EPAC showed higher and longer-lasting erythromycin levels in serum, sputum and pure bronchial mucus than ES. It is believed that EPAC is better absorbed because of its greater stability in the gastrointestinal juices. Higher concentrations in bronchial secretions not always depend on the blood levels. It seems to be possible that the N-acetylcysteine moiety in the molecule of EPAC drug can facilitate antibiotic penetration because of its mucolytic activity. The clinical response (disappearance of fever, clearance of bacterial pathogens from sputum, reduction of quantity and viscosity of sputum) also occurred faster in the EPAC than in the ES group.

Long-term multicentre trial with TA-RO CAP, a new spermicidal product.

326 healthy couples were selected to enter the trial of a new intravaginal contraceptive called the TA-RO CAP. The trial lasted three years. Clinical and laboratory examinations were performed periodically to determine the long-term effectiveness, tolerability and acceptance. Statistical evaluation of results was performed by life-table analysis.

PIP: A new vaginal cap (TA-RO-CAP) composed of multiple spermicidal contraceptive agents was tested clinically by 326 couples to evaluate the long-term effectiveness, tolerability, and acceptance of this new contraceptive method. The CAP, in the form of a watch-glass, designed for intravaginal use, contained: benzethoniom chloride, 4.5 mg; nonoxynol 9, 39 mg; polyethylene-glycol 600, 78.6 mg; polyethylene-glycol-1500, 64.4 mg; dipropylene-glycol, 11.2 mg; mannitol, 100.8 mg; starch, 40 mg; and perfume, 1.5 mg. 230 couples (70.5%) completed the trial, which involved 4582 months of use, with each couple averaging 14 months. The total pregnancies which occurred was 26; these were divided among 3 centers. 2 of 26 pregnancies were shown to result from improper usage (insertion after coitus). 29 couples discontinued use for personal reasons, and none had any side effects during the trial. 16 of these discontinuations were provoked by the male partner. 5.5% of couples experienced mild side effects, such as redness or irritation, but only 1 case discontinued because of them. None of the women showed neoplastic degeneration of the portion or in the vagina after colposcopy and vaginal oncological cytology examinations. The statistical evaluation of this 3-year long trial is presented in a life-table.