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PubMed1998

Salicylic acid induces a cytosolic Ca2+ elevation in yeast.

Cytosolic free calcium ion concentration ([Ca2+]cyt) after a salicylic acid (SA)-stimulus was monitored in cells of the yeast Saccharomyces cerevisiae expressing apoaequorin, which constitutes a Ca(2+)-sensitive luminescent protein, aequorin, when combined with coelenterazine. SA induced a transient [Ca2+]cyt elevation that was dependent on the concentration of SA and pH of the SA solution. The SA-induced [Ca2+]cyt elevation was not reduced in Ca(2+)-deficient medium, suggesting that Ca2+ was mobilized from an intracellular Ca2+ store(s). Benzoic acid, butyric acid and sorbic acid did not induced a [Ca2+]cyt elevation.

Aequorin↗
PubMed1996

In vitro endothelialization of bioprosthetic heart valves.

BACKGROUND AND AIMS OF THE STUDY: The main disadvantage of xenograft bioprosthetic heart valves is limited durability due to tissue degeneration. METHODS: To study possible alternatives to conventional bioprosthetic heart valves, glutaraldehyde preserved porcine aortic valve leaflets were precoated with fibronectin-heparin and acidic fibroblast growth factor (aFGF) to enhance cell proliferation. Furthermore, different methods of storage and preservation (1.0% benzoic acid, 1.0% sorbic acid, 0.05% and 0.5% dialdehyde starch) were compared to conventional preservation procedures. Valve leaflets were lined with adult human saphenous vein endothelial cells (1.0 x 10(4) AHSVEC/cm2). The different methods of preservation were compared to leaflets precoated additionally with 1.0 microgram/cm2 aFGF protein, derived from gene manipulated bacteria. Endothelial growth kinetics were studied and verified by an increase of activation following 3H-thymidine incorporation, while the maintained metabolic cell activity was demonstrated by prostacyclin (PGI2) release measurements. In subsequent experiments in vivo degeneration and mineralization were determined by subcutaneous implantation in rats for up to 40 days. RESULTS: Our results favored alternatively treated valve leaflets; endothelial cells grew to persistent monolayers in six to 12 days in contrast to conventional processed leaflets where no endothelial cell growth was possible. Atomic absorption spectroscopic assessment of subdermal valve implants showed significantly lower contents of calcium, magnesium and phosphate in alternatively treated leaflets. Furthermore, precoating of alternatively preserved valve leaflets with aFGF protein and endothelial cells improved in vitro and in vivo results significantly. CONCLUSIONS: Our study shows that endothelial cell growth as well as significantly reduced in vivo degeneration and mineralization of valve leaflets may be feasible if bioprosthetic heart valves are processed according to alternative, non-toxic conservation procedures and are precoated with angiogenic growth factor protein.

Adult↗
PubMed1989

Survival and growth of food poisoning bacteria following inoculation into cottage cheese varieties.

Following inoculation into cottage cheese varieties with and without sorbic acid, obtained directly from the manufacturer, strains of enteropathogenic Escherichia coli and other E. coli survived but failed to multiply during storage at 7, 10 or 25 degrees C. In the absence of sorbic acid spoilage due to Pseudomonas fluorescens occurred after storage for 5-13 days at 7 or 10 degrees C and 1-2 days at 25 degrees C. Salmonella enteritidis, S. hadar, S. saint-paul, S. typhimurium and S. virchow survived but failed to multiply at 10 degrees C and, in the case of most strains, at 20 or 25 degrees C. S. typhimurium multiplied 100-fold in one batch of cottage cheese with peppers and onion in the absence of sorbic acid during storage at 25 degrees C for 2 days; spoilage of this batch occurred due to yeasts or yeasts and moulds after storage for 4-8 days at 10 degrees C and 0-2 days at 20 or 25 degrees C. Following inoculation into cottage cheese varieties, prepared in the laboratory and which did not contain sorbic acid, as contaminants of the added protein or vegetable ingredients the numbers of Staphylococcus aureus declined during storage at 10 and 20 degrees C, the numbers of Bacillus cereus and S. typhimurium increased at both temperatures, and the numbers of Yersinia enterocolitica increased at 10 degrees C, but declined at 20 degrees C. Spoilage occurred due to the growth of moulds and P. fluorescens after storage for 5-14 days at 10 degrees C, and due to P. fluorescens after storage for up to 2 days at 20 degrees C. In products inoculated in a similar way but which contained sorbic acid (500-530 mg/kg), the numbers of S. aureus and B. cereus declined and in most products the numbers of S. typhimurium and Y. enterocolitica remained constant. In cottage cheese with chicken, however, the numbers of Y. enterocolitica increased 100-fold during storage of the product for 14 days at 10 degrees and the numbers of S. typhimurium increased 100-fold during storage for 2 days at 20 degrees C. Spoilage of this product due to P. fluorescens occurred after storage for 8-14 days at 10 degrees C, but was not evident at 20 degrees C after 2 days.

Bacillus cereus↗
PubMed2004

Prevalence of Clostridium species and behaviour of Clostridium botulinum in gnocchi, a REPFED of italian origin.

Sales and consumption of refrigerated processed foods of extended durability (REPFEDs) have increased many-fold in Europe over the last 10 years. The safety and quality of these convenient ready-to-eat foods relies on a combination of mild heat treatment and refrigerated storage, sometimes in combination with other hurdles such as mild preservative factors. The major hazard to the microbiological safety of these foods is Clostridium botulinum. This paper reports on the prevalence and behaviour of proteolytic C. botulinum and non-proteolytic C. botulinum in gnocchi, a potato-based REPFED of Italian origin. Attempts to isolate proteolytic C. botulinum and non-proteolytic C. botulinum from gnocchi and its ingredients were unsuccessful. Based on assessment of the adequacy of the methods used, it was estimated that for proteolytic C. botulinum there was < 25 spores/kg of gnocchi and < 70 spores/kg of ingredients. The total anaerobic microbial load of gnocchi and its ingredients was low, with an estimated 1 MPN/g in processed gnocchi. Most of the anaerobic flora was facultatively anaerobic. A few obligately anaerobic bacteria were isolated from gnocchi and its ingredients and belonged to different Clostridium species. The protection factor, number of decimal reductions in the probability of toxigenesis from a single spore, was determined for eight different gnocchi formulations by challenge test studies. For all gnocchi stored at 8 degrees C (as recommended by the manufacturer) or 12 degrees C (mild temperature abuse), growth and toxin production were not detected in 75 days. The protection factor was >4.2 for proteolytic C. botulinum, and >6.2 for non-proteolytic C. botulinum. When inoculated packs were stored at 20 degrees C (severe temperature abuse), toxin production in 75 days was prevented by the inclusion of 0.09% (w/w) sorbic acid (protection factors as above), however in the absence of sorbic acid the packs became toxic before the end of the intended shelf-life and the protection factors were lower. Providing sorbic acid (0.09% w/w) is included in the gnocchi, the safety margin would seem to be very large with respect to the foodborne botulism hazard.

Ascorbic Acid↗
PubMed1997

trans,trans-muconic acid as a biomarker of non-occupational environmental exposure to benzene.

Excretion of trans,trans-muconic acid (2,4-hexadienedioic acid; t,t-MA), a potential biomarker of low-level exposure to benzene, was determined in 32 smokers and 82 nonsmokers. In smokers the median background excretion of t,t-MA was 0.13 (0.06-0.39) mg/g creatinine and was significantly higher (P < 0.05) than the value of 0.065 (0.02-0.59) mg g creatinine in nonsmokers. For nonsmokers, the correlation between t,t-MA excretion and environmental exposure to benzene in ambient air, which was determined during the 8-day study period by personal diffusion samplers, was not significant (r = 0.164, P = 0.18). Nonsmokers living in the city tended to have higher t,t-MA excretion rates than nonsmokers living in the suburbs. However, the difference was only significant for nonsmokers from nonsmoking homes. For the same location (suburb or city), smoking at home leads to a marginal increase in t,t-MA excretion of the nonsmoking members of the household. In a further study with eight nonsmokers we found that dietary supplementation with 500 mg sorbic acid significantly increased (P < 0.001) the mean urinary t,t-MA excretion from 0.08 (0.04-0.12) to 0.88 (0.57-1.48) mg/24 h. Under study conditions 0.12% of the sorbic acid dose was excreted in urine as t,t-MA, thereby indicating that a typical dietary intake of 6-30 mg/day sorbic acid accounts for 10-50% of the background t,t-MA excretion in nonsmokers, and for 5-25% in smokers. As a consequence, sorbic acid in the diet is a significant confounding factor in assessing low-level benzene exposure if t,t-MA excretion in urine is used as a biomarker.

Benzene↗
PubMed1985

Multiple modes of inhibition of spore germination and outgrowth by reduced pH and sorbate.

Germination and outgrowth of three strains of Clostridium botulinum in PYEG medium were measured by phase contrast microscopy. Reduction in pH from 7 to 5.5 completely inhibited germination of strain 12885A, reduced the extent of germination of strain 62A and had no effect on the extent of germination of strain 53B. At pH 5.5, 225 mg/l of undissociated sorbic acid had no effect on the germination of strain 53B, while at pH 6.5, 225 mg/l of undissociated sorbic acid completely inhibited germination of strains 62A and 12885A. Outgrowth of germinated spores of strains 62A and 53B was not inhibited at pH 5.5, but the addition of sorbate (225 mg/l undissociated sorbic acid) completely inhibited outgrowth. Sorbate inhibited germination of Cl. botulinum and Bacillus cereus spores triggered to germinate by amino acids. Inhibition occurred after germinant binding, as measured by commitment to germinate.

Bacillus cereus↗
PubMed2000

Simultaneous determination of sweeteners and preservatives in preserved fruits by micellar electrokinetic capillary chromatography.

A micellar electrokinetic capillary method for the simultaneous determination of the sweeteners dulcin, aspartame, saccharin, and acesulfame-K and the preservatives sorbic acid; benzoic acid; sodium dehydroacetate; and methyl-, ethyl-, propyl-, isopropyl-, butyl-, and isobutyl-p-hydroxybenzoate in preserved fruits is developed. These additives are ion-paired and extracted using sonication followed by solid-phase extraction from the sample. Separation is achieved using a 57-cm fused-silica capillary with a buffer comprised of 0.05 M sodium deoxycholate, 0.02 M borate-phosphate buffer (pH 8.6), and 5% acetonitrile, and the wavelength for detection is 214 nm. The average recovery rate for all sweeteners and preservatives is approximately 90% with good reproducibility, and the detection limits range from 10 to 25 microg/g. Fifty preserved fruit samples are analyzed for the content of sweeteners and preservatives. The sweeteners found in 28 samples was aspartame (0.17-11.59 g/kg) or saccharin (0.09-5.64 g/kg). Benzoic acid (0.02-1.72 g/kg) and sorbic acid (0.27-1.15 g/kg) were found as preservatives in 29 samples.

Acetonitriles↗
PubMed2003

trans,trans-Muconic acid excretion in relation to environmental exposure to benzene.

OBJECTIVES: Potential environmental sources of benzene exposure, and intake of foods and beverages susceptible to being preserved with sorbic acid, were investigated in relation to their contribution to the inter-individual variation in background urinary trans,trans-muconic acid ( t,t-MA) excretion among subjects non-occupationally exposed to benzene. METHODS: We measured urinary t,t-MA excretion in 65 subjects, 34 women and 31 men. A spot sample of morning urine was collected for each subject 10-12 h after they had consumed their last meal. Questionnaire information was collected on diet and possible sources of environmental benzene exposure in the surroundings of the subjects' residences. For each subject, an estimate of the average daily intake of sorbic acid with diet was calculated, based on questionnaire information and laboratory data on samples of local food items. RESULTS: The t,t-MA geometric mean was significantly higher among women (28.7 vs 11.5 microg/g creatinine, P<0.05) and among smokers (37.6 vs 15.6 microg/g creatinine, P<0.05), and increased by years of education among women, but not among men. In the multivariate analysis, smoking was the only significant predictor of elevated t,t-MA excretion. In our study, the average estimated daily sorbic-acid intake with diet was 0.33 ppm (standard deviation: 0.28), and it did not show a correlation with t,t-MA excretion. Urban traffic and residence within 100 m of a fuel station also did not show an association with elevated t,t-MA values. CONCLUSIONS: Our study confirms that, among subjects non-occupationally exposed to benzene, smoking contributes significantly to increased background t,t-MA excretion. Further studies should be addressed to confirm our observation of elevated t,t-MA levels among women.

Adult↗
PubMed2001

Prevalence and behaviour of Bacillus cereus in a REPFED of Italian origin.

This paper reports on the prevalence and behaviour of Bacillus cereus in gnocchi, a REPFED of Italian origin. A survey of gnocchi under varying storage conditions, revealed that, although B. cereus was found in 33% of the samples, the contamination level was lower than 10(2) CFU/g for the unstored and the refrigerated (8 degrees C) samples. Counts increased with increasing storage temperatures and prolonged storage times in samples prepared without sorbic acid. The effect of different formulations (sorbic, citric and lactic acid) and storage conditions (8, 12 and 20 degrees C) have been evaluated in a challenge testing with spores of B. cereus. Results indicate that the use of sorbic acid in association with citric or lactic acid to pH 5.0 is effective in inhibiting growth of B. cereus and the anticipated shelf life of the product is safe even if temperature abuse occurs. If sorbic acid is omitted, lactic acid can inhibit B. cereus growth during storage at 8 degrees C. On the contrary, when temperature abuse occurs (12 and 20 degrees C), lactic or citric acid are not able to prevent growth of B. cereus.

Bacillus cereus↗
PubMed2000

Positive and negative control of multidrug resistance by the Sit4 protein phosphatase in Kluyveromyces lactis.

The nuclear gene encoding the Sit4 protein phosphatase was identified in the budding yeast Kluyveromyces lactis. K. lactis cells carrying a disrupted sit4 allele are resistant to oligomycin, antimycin, ketoconazole, and econazole but hypersensitive to paromomycin, sorbic acid, and 4-nitroquinoline-N-oxide (4-NQO). Overexpression of SIT4 leads to an elevation in resistance to paromomycin and to lesser extent tolerance to sorbic acid, but it has no detectable effect on resistance to 4-NQO. These observations suggest that the Sit4 protein phosphatase has a broad role in modulating multidrug resistance in K. lactis. Expression or activity of a membrane transporter specific for paromomycin and the ABC pumps responsible for 4-NQO and sorbic acid would be positively regulated by Sit4p. In contrast, the function of a Pdr5-type transporter responsible for ketoconazole and econazole extrusion, and probably also for efflux of oligomycin and antimycin, is likely to be negatively regulated by the phosphatase. Drug resistance of sit4 mutants was shown to be mediated by ABC transporters as efflux of the anionic fluorescent dye rhodamine 6G, a substrate for the Pdr5-type pump, is markedly increased in sit4 mutants in an energy-dependent and FK506-sensitive manner.

4-Nitroquinoline-1-oxide↗
PubMedDate not supplied

Mechanisms of subjective (sensory) irritation. Propensity to non-immunologic contact urticaria and objective irritation in stingers.

Subjective (sensory) irritation was studied with lactic acid test on 74 females; 8 reacted positively with "stinging" and/or itching or burning and another 5 had mild subjective sensations. History of previous corresponding sensations from contact with cosmetic products was significantly more common in stingers than in non-stingers (p less than 0.001); other factors, such as dermatologic history, sensitivity to ultraviolet light or skin dryness had no characteristic patterns. In the objective, immediate non-immunologic contact urticaria tests with sorbic acid and benzoic acid, the stingers developed significantly more erythema to 0.5% sorbic acid (p less than 0.05) and to 1% benzoic acid (p less than 0.02). 1% sorbic acid also induced more edema in stingers (p less than 0.02). Increased reactivity to a 24 h sodium lauryl sulfate (SLS) patch test was demonstrated only with laser Doppler velocimetry (p less than 0.05). Stingers and non-stingers reacted similarly to open, cumulative SLS irritation as measured with transepidermal water loss. These studies provide clinical and biological information about subjective irritation; the data suggest a functional correlation for the what had been considered a purely subjective (sensory) phenomena.

Adolescent↗
PubMed1985

Potential for Development of Tolerance by Penicillium digitatum and Penicillium italicum after Repeated Exposure to Potassium Sorbate.

Two strains of Penicillium digitatum and one strain of Penicillium italicum were exposed to various levels of sorbic acid and potassium sorbate, and the MICs were determined. Selected strains of the molds were then repeatedly exposed to subinhibitory levels of the compounds to determine whether increased tolerance might develop. The MIC of sorbic acid (pH 4.75) to P. digitatum was between 0.02 and 0.025%. The MIC of sorbate (pH 5.5) to two strains of P. digitatum and P. italicum was found to be between 0.06 and 0.08%. Increasing levels of sorbate resulted in increasing growth suppression of the molds. Populations of P. digitatum were tested for increased tolerance to sorbic acid, and none was found. Individual molds that started from the same parent colony were examined for increased tolerance to potassium sorbate. Two P. digitatum strains developed no observable increased tolerance, but P. italicum developed a slight increase in tolerance to sorbate. When spores of P. italicum and P. digitatum were exposed to higher levels of sorbate for prolonged times, the fungicidal or fungistatic activity of the inhibitor was dependent upon pH, length of exposure time, level of sorbate, and the mold strain.

Journal Article↗
PubMed2006

Use of experimental design methodology to prepare Maillard reaction products from glucose and cysteine inhibitors of polyphenol oxidase from eggplant (Solanum melongena).

Polyphenol oxidase (PPO) from eggplant was extracted and partially purified by a two-step fractionation-precipitation using ammonium sulfate and phenylsepharose hydrophobic interaction chromatography. The eggplant PPO extract was characterized concerning its kinetic properties. Optimal conditions to obtain Maillard reaction products (MRPs) with a maximal inhibitory potency (IP) toward PPO activity were determined using the surface response methodology and a four-factor and five-level experimental design. The MRPs were prepared from cysteine (0.25 M) and glucose (0-1 M), at several initial pH values (2-6) and at differing heating times (3-19 h) and temperatures (95-115 degrees C). The maximal IP was obtained after heating a model system of glucose/cysteine (1/0.25 M) at pH 2 for 3 h 20 min at 115 degrees C. The soluble part of this MRP, called MRP(IPmax), was a noncompetitive inhibitor toward eggplant PPO. The IP of MRP(IPmax) on PPO activity was very potent as compared to that displayed by benzoic, p-coumaric, and t-cinnamic acids, as well as sorbic acid and 4-hexylresorcinol. The activity of preincubated PPO at 0 degrees C with MRP(IPmax) was only slightly restored after dialysis or gel filtration.

Benzoic Acid↗
PubMedDate not supplied

Simultaneous gas chromatographic determination of carboxylic acids in soft drinks and jams.

A method was developed for simultaneous gas chromatographic determination of sorbic acid, dehydroacetic acid, and benzoic acid used as preservatives, and succinic acid, fumaric acid, malic acid, and tartaric acid used as acidulants in soft drinks and jams. A sample was dissolved in NH4OH-NH4Cl pH 9 buffer solution, and an aliquot of the solution was passed through a QAE-Sephadex A 25 column. The column was washed with water, and the carboxylic acids were eluted with 0.1N HCl. Sorbic acid, dehydroacetic acid, and benzoic acid were extracted with ethyl ether-petroleum ether (1 + 1), and determined on a 5% DEGS + 1% H3PO4 column. Succinic acid, fumaric acid, malic acid, and tartaric acid in the lower layer were derivatized with N,O-bis(trimethylsilyl)acetamide and trimethylchlorosilane, and determined on a 3% SE-30 column. Recoveries from soft drink and jam samples fortified with 0.1% each of 7 carboxylic acids ranged from 92.4 to 102.6% for preservatives, and from 88.1 to 103.2% for acidulants.

Beverages↗
PubMed1989

The response of Gluconobacter oxydans to sorbic and benzoic acids.

The minimal inhibitory concentrations (MIC) of sorbic and benzoic acids for Gluconobacter oxydans were 1000 mg/l and 900 mg/l respectively at pH 3.8. A reduction in the pH of the test medium to 3.3 reduced the MIC of both preservatives by about 300 mg/l. When G. oxydans was grown in the presence of sublethal concentrations of sorbic or benzoic acids before the MIC was determined, the MIC of both compounds increased substantially within 1 h. Growth of G. oxydans was modified in several ways by the presence of sorbic acid in the medium. The duration of the lag phase increased and there was a substantial decrease in the viable count during the lag phase in the presence of high concentrations. The generation time increased and the viable count at the end of the logarithmic phase was reduced. At 1 degree C, G. oxydans grew in the absence of sorbic acid but was inactivated by 400 mg sorbic acid/l. At 37 degrees C the viable count of suspensions of G. oxydans decreased in both the absence and presence of sorbic acid. Sorbic acid increased the death rate. Growth of G. oxydans was prevented by eliminating air from culture vessels, combined with the addition of ascorbate to the medium containing 400 mg sorbic acid/l.

Aerobiosis↗