Supercritical fluid extraction of hydrocarbons and 2-alkylcyclobutanones for the detection of irradiated foodstuffs.

Supercritical carbon dioxide can be used to carry out a selective and fast extraction (30 min) of volatile hydrocarbons and 2-alkylcyclobutanones contained in irradiated foods. After elimination of the traces of triglycerides still contained in the extracts on a silica column, the compounds were analysed by gas chromatography-mass spectroscopy (2-alkylcyclobutanones) and gas chromatography-flame ionization detection (volatile hydrocarbons). The present method was applied successfully to freeze-dried samples (1 g or less) of cheese, chicken, avocados and to various ingredients (chocolate, liquid whole eggs) included in non-irradiated cookies. It was faster (4-5 h) than the reference methods EN 1784 (volatile hydrocarbons) and EN 1785 (2-alkylcyclobutanones), which take 1.5 days each. The minimal dose detectable by this method is, in addition, slightly lower than those of the reference methods.

Determination of folates in foods by high-performance liquid chromatography with fluorescence detection after precolumn conversion to 5-methyltetrahydrofolates.

A liquid chromatographic-fluorimetric determination of folates in foodstuffs including their extraction, without or with deconjugation, chemical conversion to 5-CH3-H4PteGlu(n) and purification of the extract by affinity chromatography is reported. The conversion enables the analysis of total folates and also of the contents of the different mono- and polyglutamate forms of the folates. The method has a satisfactory day-to-day repeatability (never,more than 10%) and a very low detection limit (0.02 pmol per injection). Depending on the folate studied, the recovery rates varied from 78% (10-CHO-PteGlu) to 98% (5-CHO-H4PteGlu). Furthermore it has been possible to show that the deconjugation of the folates by rat plasma conjugase was incomplete in foodstuffs whereas chicken pancreas conjugase effectively converted the different folate polyglutamates into folate diglutamates. It could not be demonstrated that prior hydrolysis with a protease and amylase was useful for the analysis of the different foodstuffs studied (yeast, spinach, beef liver, beef fillet and peas) when deconjugation was performed with the chicken pancreas conjugase.

Fluorimetric determination of pantothenic acid in foods by liquid chromatography with post-column derivatization.

A method to determine the content of free pantothenic acid in various foods by reverse phase liquid chromatography-fluorimetry is reported. It includes a purification of the samples by successive passages through anion and cation exchange cartridges and a post-column derivatization of pantothenic acid as the fluorescent 1-alkylthio-2-alkylisoindole (reaction of beta-alanin, formed by hot alkaline hydrolysis of pantothenic acid, with orthophthaldialdehyde in the presence of 3-mercaptopropionic acid). An enzymatic hydrolysis prior to the purification step (pepsin at 50 degrees C for 3 h, then pantetheinase and alkaline phosphatase at 20 degrees C for 18 h) made it possible to release the bound pantothenic acid and thus to obtain the total Vitamin B5 content of these foodstuffs. The method proposed for the determination of free and bound pantothenic acid gives a good recovery rate (96-101%) and a satisfactory repeatability (R.S.D.r less than 8%). Owing to its low detection limit (0.65 microg g(-1)) and the good resolution of the pantothenic acid peak, it could most probably be applied to the determination of this vitamin in any foodstuff.

2-Alkylcyclobutanones as markers for irradiated foodstuffs. III. Improvement of the field of application on the EN 1785 method by using silver ion chromatography.

The inclusion of a purification step by silver ion chromatography in the EN 1785 analytical protocol for 2-alkylcyclobutanones (validated by the European Committee for Standardization for the detection of ionizing radiation treatment) has considerably improved the quality of the chromatograms obtained, allowing the detection of food samples irradiated at very low doses (0.1 kGy) or irradiated ingredients included in low proportions in non irradiated foodstuffs. This analytical modification of the protocol EN 1785 ought thus to permit a very considerable extension of its current field of application.

Inhibition of Listeria monocytogenes in a smear-surface soft cheese by Lactobacillus plantarum WHE 92, a pediocin AcH producer.

The anti-Listeria monocytogenes activity of Lactobacillus plantarum WHE 92, a pediocin AcH producer, was investigated in Munster cheese, a smear-surface soft cheese. The appearance of L. monocytogenes in the cheese, which naturally occurs solely in the crust and never before 1 week of ripening, could be prevented by spraying a cell suspension of L. plantarum WHE 92 (ca. 10(5) CFU/ml) on the cheese surface at the beginning of the ripening period. L. monocytogenes was sometimes detected at low levels (<5.0 x 10(1) CFU/g) after 7 to 11 days of ripening. However, this pathogen not be able to grow, nor did it survive the presence of L. plantarum WHE 92 in any of the samples examined until the end of the ripening not be able to grow, nor did it survive the presence of than 10(4) CFU/g in control samples. In other respects, L. plantarum WHE 92, which exists naturally in Munster cheese, did not adversely affect the evolution of the ripening process. This procedure has allowed manufacturers to successfully put an antilisterial treatment into practice in their ripening rooms.

[Aflatoxins in food: tests of decontamination of peanut cakes by ionizing treatment]. / Aflatoxines dans les aliments: essais de décontamination des tourteaux d'arachide par traitement ionisant.

The efficacy of ionising treatment for decontaminating peanut cakes was tested. The influence of cakes water content and the effect of ionisation dose rate were studied. The results obtained after a reverse phase liquid chromatographic determination of B1, B2, G1 and G2 aflatoxins have revealed an important contamination of the peanut cakes (up to 1000 ppb of total aflatoxin's contents). After ionising treatment at 25 kGy, the aflatoxins degradation in peanut cake's was less important in dried samples (about 5-10% at 0.55 water activity: aw) than in the humid ones (40-60% degradation at 0.95 water activity). At this dose, any indicative difference of the degradation rate of aflatoxins, with regard to the ionising process was observed. The efficacy of ionising treatment for decontaminating peanut cakes could probably be improved, however the economic interest of such process as alternative of the treatment with ammonia is questionable.

Continuous subcutaneous insulin infusion allows tolerance induction and diabetes treatment in a type 1 diabetic child with insulin allergy.

AIM: Insulin allergy is a rare but serious and challenging condition in patients with type 1 diabetes (T1D). This is a case report of an 8-year-old boy with T1D and an allergy to insulin. CASE REPORT: Three months after being diagnosed with T1D, the patient developed progressive skin reactions to insulin, characterized by small 1.5-cm pruritic wheals at injection sites that persisted for several days. Seven months after diagnosis, he experienced two episodes of generalized urticaria with systemic symptoms that were seen within a few seconds of insulin injection. Examination revealed lipoatrophy of the thighs. Intradermal skin tests were positive for protamine, glargine and lispro. The patient was started on a continuous subcutaneous insulin infusion (CSII) tolerance induction protocol, consisting of a very low basal rate that was progressively increased, with the first bolus given under medical supervision, and was well tolerated for 4 months. After this period of time, the skin wheals reappeared, localized to the infusion sites, but without urticaria or any other generalized reactions. Intradermal skin tests were repeated and were again positive. Serum insulin-specific IgE measured 30 months after the first allergic reactions were positive. After 3 years, pump therapy is ongoing and blood glucose control has remained relatively good (HbA1c 7.6%). CONCLUSION: In T1D children with insulin allergy, CSII can successfully be used to both induce insulin tolerance and allow diabetes insulin therapy, although insulin desensitization cannot always be fully achieved. The induction protocol was easily manageable partly due to the "honeymoon" period that the patient was still in, but it should nonetheless be used even when the patient has higher insulin requirements.

[Benefits of continuous subcutaneous insulin infusion in children with type 1 diabetes mellitus]. / Bénéfices de l'insulinothérapie par pompe chez les enfants diabétiques de type 1.

BACKGROUND: The Diabetes Control and Complications Trial clearly demonstrated the benefits of blood glucose control, especially in children and adolescents, in the prevention of long-term complications of type 1 diabetes (T1D). This can be achieved with intensive insulin treatment with either multiple daily insulin injections (MDI) or continuous subcutaneous insulin infusion (CSII), also known as insulin pump. The aim of this study was to compare glycemic control of T1D children treated with either CSII or MDI. PATIENTS AND METHODS: Thirty-eight T1D children treated with CSII were compared to 38 children treated with MDI, matched for age, gender, and duration of diabetes. Collected data, including daily doses of insulin in IU/kg/d, HbA1c levels, body mass index expressed in standard deviation/age, number of severe hypoglycemia episodes and of admissions related to T1D expressed in events/patient/year, were retrospectively collected every 3 months. RESULTS: There was no difference between the 2 groups at baseline. During the 3 years of follow-up, patients treated with CSII had lower daily doses of insulin (0.78 ± 0.19 vs. 0.87 ± 0.22 IU/kg/d, p<0.05), significantly lower levels of HbA1c (7.5 ± 0.6 vs. 8.0 ± 1.3 %, p<0.05), and a decreased number of admissions related to T1D (0.07 ± 0.14 vs. 0.17 ± 0.22 events/patient/year, p<0.05) than children treated with MDI. In contrast, body mass index and number of severe hypoglycemic episodes did not differ between the two groups. No diabetic ketoacidosis episode was recorded in either group. CONCLUSION: The results from this study suggest that treatment with CSII provided better metabolic control than treatment with MDI, in spite of lower daily doses of insulin and without increasing acute complications, in children with T1D.

Impaired neuromuscular transmission after recovery of the train-of-four ratio.

BACKGROUND: Residual neuromuscular blockade may increase the risk of development of post-operative pulmonary complications, but is difficult to detect clinically. It was speculated that patients may have impaired neuromuscular transmission after surgery of long duration, despite the recovery of the train-of-four (TOF) ratio. METHODS: The muscle force (mechanomyography), motor compound muscle action potential amplitude and fatigue of the adductor pollicis (AP) muscle were assessed after recovery of the TOF ratio to 0.9. Thirteen patients receiving repetitive administration of neuromuscular blocking agents (NMBAs) during surgery (median, 5.3 h; interquartile range, 3.4-6 h) were studied post-operatively in the intensive care unit. At the time of the measurements, patients were scheduled for extubation and the AP TOF ratio amounted to a mean (standard deviation, SD) of 0.94 (0.05). Six healthy volunteers of similar age, weight and gender were studied for comparison. Force-frequency curves were generated by stimulation (10-80 Hz) of the ulnar nerve, and the AP electromyogram (EMG) amplitude was measured, in parallel, before and after evoked muscle fatigue. RESULTS: The maximum AP force at a stimulation frequency of 20-80 Hz was significantly lower in patients than in controls [40 N (16 N) vs. 65 N (18 N) at 80 Hz]. In patients, but not in controls, the EMG amplitude decreased with increasing nerve stimulation frequency, and a tetanic fade of both force and EMG, amounting to 0.41 (0.33) (EMG) and 0.61 (0.35) (mechanomyography) at 80 Hz, was observed. Force after fatiguing contractions did not differ between the groups. CONCLUSION: After repetitive administration of NMBAs during surgery, even with recovery of the TOF ratio to 0.9 or more, muscle weakness from impaired neuromuscular transmission can occur. The clinician should consider that post-operative recovery of the TOF ratio to 0.9 does not exclude an impairment of neuromuscular transmission.

Antilisterial activity of enterocin 81, a bacteriocin produced by Enterococcus faecium WHE 81 isolated from cheese.

Enterocin 81, a bacteriocin produced by Enterococcus faecium WHE 81 previously isolated from cheese, exhibited a very narrow spectrum of activity, which is mainly directed against enterococci and Listeria spp. including Listeria monocytogenes. Enterocin 81 activity, which was extremely rapid with maximal effect achieved within 30 min, could not be detected after treatment with various proteolytic enzymes. This activity was bactericidal in nature and induced an important efflux of intracellular material, which was visualized under electron microscopy as filaments coming out of L. monocytogenes cells. However, enterocin 81 did not display bacterial lysis on sensitive cells, as no changes in cell morphology were detected following the bactericidal action. Furthermore, this bacteriocin was shown to be equally active at pH values ranging from 4.0 to 8.0, which, along with the narrow activity spectrum, are two factors of paramount interest with regards to possible use of this bacteriocin in fermented foods.

[Residue analysis of methiocarb, repellent for the Quelea, in crops var. sorghum sp. (author's transl)]. / Analyse dans les graminées de variété sorghum sp. des résidus d'un répulsif pour le Quelea: le méthiocarb.

The methiocarb (4-methylthio 3,5-dimethylphenyl N-methylcarbamate) has repellent properties towards against Quelea, a small bird deteriorating crops in Africa. The residue analysis of the carbamate compound and of its possible metabolites on treated Sorghum sp. was completed by TLC after chloroformic quantitative extraction. The isolation of metabolites by two dimensional thin layer chromatography demonstrates the presence of methiocarb sulfone. The quantitative survey of residues was achieved by the treatment of the methiocarb by hydrolyzed derivatives by dansylchloride, a very fluorescent component. The fluorescent densitometry of dansylated by methylamine allows a total measurement of the residues at a level of less than 10 ng. This investigation reveals the disappearance curve of methiocarb in our samples in process of time and to define the half-live and, therefore, the intervals of treatment. Taking into account the RI 50 index, our results provide encouraging support for the utilization of methiocarb in the eradication of the harmful Quelea.

Production of pediocin AcH by Lactobacillus plantarum WHE 92 isolated from cheese.

Among 1,962 bacterial isolates from a smear-surface soft cheese (Munster cheese) screened for activity against Listeria monocytogenes, six produced antilisterial compounds other than organic acids. The bacterial strain WHE 92, which displayed the strongest antilisterial effect, was identified at the DNA level as Lactobacillus plantarum. The proteinaceous nature, narrow inhibitory spectrum, and bactericidal mode of action of the antilisterial compound produced by this bacterium suggested that it was a bacteriocin. Purification to homogeneity and sequencing of this bacteriocin showed that it was a 4.6-kDa, 44-amino-acid peptide, the primary structure of which was identical to that of pediocin AcH produced by different Pediococcus acidilactici strains. We report the first case of the same bacteriocin appearing naturally with bacteria of different genera. Whereas the production of pediocin AcH from P. acidilactici H was considerably reduced when the final pH of the medium exceeded 5.0, no reduction in the production of pediocin AcH from L. plantarum WHE 92 was observed when the pH of the medium was up to 6.0. This fact is important from an industrial angle. As the pH of dairy products is often higher than 5.0, L. plantarum WHE 92, which develops particularly well in cheeses, could constitute an effective means of biological combat against L. monocytogenes in this type of foodstuff.