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Indirect food additive migration from polymeric food packaging materials.

Many foods contact polymeric packaging materials which contain residues of the polymerization process or additives employed to facilitate processing. The extent of migration of such materials from the packaging to foods is the focus of the present article. A major experimental program using eight polymer-migrant systems is described. Migration was measured to food-simulating liquids (FSL) and to foods. Accelerated tests were conducted with FSL under FDA guidelines conditions so as to develop correlations between such data and those found using foods under normal storage temperatures and shelf lives. In the majority of tests, the migration was found to be approximately proportional to the square root of time, to increase significantly with a rise in temperature, and to be proportional to the initial concentration of migrant in the polymer. Stirring in the FSL or food phase was generally not important except for the system involving dioctyl adipate migrating from polyvinyl chloride film. In some instances, after a period of time, migration rates became very low, and this effect was attributed to saturating the FSL or food phase with migrant. The foods comprised a variety of types, including liquid, semisolid, solid, and dry; both oily and aqueous foods were included. The physical steps involved in migration include the diffusion of the migrant from the interior of the film to the surface, where it can dissolve in the external FSL or food phase. The nature of the FSL or food is shown to be very important in that components can penetrate the polymer and dramatically increase migration rates. Consistent with the FDA guidelines in effect at the time of this study, testing was performed with five FSL (water, 3% acetic acid, 8% and 50% ethanol, and n-heptane) at 49 degrees C. Detailed comparisons were made between the migrations to foods and to FSL; following are the more relevant conclusions. (1) Three percent acetic acid showed no advantage over water as a food simulant even in those cases where the food could be considered acidic in nature. (2) Water, when used as an FSL at 49 degrees C for 5 days, overestimated migration in aqueous foods in about 75% of the cases. In some instances, however, the water phase became saturated with migrant. In other situations, this test protocol underpredicted migration--especially in those cases where there were components in the food that were able to penetrate into the polymer and enhance migration (such as orange juice).(ABSTRACT TRUNCATED AT 400 WORDS)

Chemical Phenomena↗

Properties of cholesterol-reduced butter made with beta-cyclodextrin and added evening primrose oil and phytosterols.

The present study was carried out to examine changes in the chemical and sensory properties of butter in which the cholesterol was reduced and to which evening primrose oil (EPO) and phytosterols were added. Crosslinked beta-cyclodextrin (beta-CD) made from adipic acid was used, and approximately 90% of the cholesterol was removed. The color measurement values "L" and "a" were significantly different between the control (butter with no beta-CD treatment and no added EPO and phytosterols) and treatment A (butter treated with 10% crosslinked beta-CD); however, the color values for "L" and "a" were similar. The color value "b" in treatment B (butter treated with 10% crosslinked beta-CD and 5% phytosterols and 3% EPO added) was significantly higher than in the other treatments. The thiobarbituric acid value of treatment B was significantly higher than that of the control and treatment A. Scores for hardness, elasticity, and cohesiveness were significantly lower in the control than in treatment A. Differences in sensory characteristics did not result from the beta-CD treatment but from the addition of EPO and phytosterols. In microscopic examinations, no noticeable differences were found among the treatments, and a smooth texture and a fine, uniform crystalline structure were observed. Results indicated that about 90% of the cholesterol was reduced by crosslinked beta-CD and that the beta-CD treatment itself did not adversely influence the chemical and sensory properties of the butter. However, the addition of EPO and phytosterols to the butter appeared to impair its sensory properties, especially in terms of rancidity and overall acceptability.

Butter↗

Method for the routine quantitative gas chromatographic analysis of major free fatty acids in butter and cream.

A rapid quantiative method was developed for routine analysis of the major, even carbon-numbered free fatty acids in butter and cream. Free fatty acids were isolated directly from intact samples by a modified silicic acid-potassium hydroxide arrestant column and were separated by gas chromatography with a 1.8 m x 2 mm inner diameter glass column packed with 10% neopentyl glycol adipate on 80/100 Chromosorb W. Purified, formic acid-saturated carrier gas was required for minimal peak tailing and extended column life. The accuracy and reproducibility of the mmethod was established through quantitative recovery studies of free fatty acid mixtures, free fatty acids added to butter, and replicate analysis of butter and cream samples.

Chromatography, Gas↗

Effect of vitamin B6 deficiency on collagen metabolism in rats.

The effects of VB6 deficiency on collagen metabolism were studied using young and adult rats. It was observed in VB6 deficient rats, that both the amount of urinary hydroxyproline and salt soluble fraction of skin collagen decreased. Furthermore, decrease of the alpha-component and increase of the beta-component were also found in the soluble collagen. However, the activity of plasma amine oxidase was lower and the aldehyde content in the acid soluble collagen were also lower than the control group. These changes were revealed more clearly in young growing rats than in adult rats. Based on these results, it may be that VB6 participates in the allysine (alpha-amino-adipic-delta-semialdehyde) formation which is considered as the first step of collagen maturation and also in the synthesis of protocollagen peptide chains.

2-Aminoadipic Acid↗

[Identification of unknown substances in polyvinyl chloride gloves containing non-phthalate plasticizers].

The use of polyvinyl chloride gloves containing di(2-ethylhexyl) phthalate for food contact applications is restricted. In their place, polyvinyl chloride gloves containing non-phthalate plasticizers (PVC-NP) have been introduced. They contained unknown substances, so they were studied by GC/MS, HR-MS and NMR. The chemical structures of the unknown substances were confirmed to be diethylene glycol dibenzoate, triethylene glycol dibenzoate, dipropylene glycol dibenzoate, and alkylsulfonic acid phenyl ester, which are plasticizers. Including di-isononyl adipate, the contents of the plasticizers were 37.5-48.9% in the PVC-NP gloves, and their migration levels were 1,010-1,390 ppm into n-heptane. These are very high levels. These plasticizers are not widely used for food contact polyvinyl chloride throughout the world, and they have also not been registered as self-standards by Japanese manufacturers. Careful consideration will be necessary for the selection of a suitable plasticizer substitute.

Gas Chromatography-Mass Spectrometry↗

[Contents of plasticizers in cap-sealing for bottled food and their migration into food].

Plasticizers in cap-sealing for bottled foods were analyzed. Twenty-three domestic samples and 80 imported samples, a total of 103 samples, were tested. Among them, 93 contained chloride, and 62 contained di(2-ethylhexyl) phthalate (DEHP), diisodecyl phthalate, O-acetyl tributyl citrate, diacetyllauroyl glycerol, di(2-ethylhexyl) adipate, diisononyl phthalate or dicyclohexyl phthalate. Twelve samples with DEHP-containing caps were further tested for plasticizer levels in the foods. Higher DEHP was detected in oily and fluid food specimens. However, the intake of plasticizers calculated based on usual food consumption did not exceed the tolerable daily intake level in any of the cases. A food sample containing the highest level of DEHP was stored under various conditions, and DEHP that migrated from the cap-sealing into the food was determined. Shaking the bottles increased migration of DEHP into foods.

Diethylhexyl Phthalate↗

A rare case of enteropathy-associated T-cell lymphoma presenting as acute renal failure.

Enteropathy-associated T-cell lymphoma (EATCL) is a high grade, pleomorphic peripheral T-cell lymphoma usually with cytotoxic phenotypes. We describe a first case of patient with EATCL that is remarkable for its fulminant course and invasion of both kidneys manifested as acute renal failure. The patient was a 23 year old woman with a long history of celiac disease. She was presented with acute renal failure and enlarged mononuclear infiltrated kidneys. Diagnosis of tubulointerstitial nephritis and polyserositis was confirmed with consecutive pulse doses of steroid therapy. After recovery, she had disseminated disease two months later. Magnetic resonance imaging showed thickened intestine wall, extremely augmented kidneys, enlarged intra-abdominal lymph nodes with extra-luminal compression of common bile duct. Laparotomy with mesenterial adipous tissue and lymph glands biopsy was done. Consecutive pathophysiological and immunohistochemical analyses confirmed the diagnosis of EATCL: CD45RO+, CD43+, CD3+. The revision of renal pathophysiology substantiated the diagnosis. The patient received chemotherapy, but unfortunately she died manifesting signs of pulmonary embolism caused by tumor cells.

Acute Kidney Injury↗

Toxicity of rubber chemicals towards three-day chicken embryos.

Three-day chicken embryos were exposed to 80 different rubber chemicals dissolved in either acetone or water. The following classes of chemicals were studied: thiurams, dithiocarbamates, thioureas, benzothiazoles, benzenesulfohydrazide, dithiodimorpholine, amines, acrylates, guanidines, resorcinol, phthalates, adipates, phosphates, oils, peroxides, heavy metal salts, and sodium nitrite. Dose-response curves and median lethal doses (LD50) or median effective doses (ED50) were calculated for mortality in 2 d after the treatment, for total mortality in 11 d after the treatment, and for the total effect, including deaths and malformations 11 d after the treatment, when the test was finished. Sixteen of the chemicals had no effect on the embryos when injected into the air chamber. Incomplete and irregular dose-response curves for the total effect were obtained with 13 of the chemicals. For them, the ED50 could be calculated. Among the remaining 51 chemicals, the most potent were the dithiocarbamates and cadmium and copper acetates, with the total effect ED50 from 2.4 to 160 nmol/egg. Other chemicals that had an ED50 smaller than 100 nmol/egg were thiurams, cyclohexylthiophthalimide, acrolein, and dithiodimorpholine. The majority of the chemicals had an ED50 between 100 nmol/egg and 10 mumol/egg. The least potent were sodium nitrite and methylmethacrylate with an ED50 of 22 mumol/egg. All the 64 embryotoxic chemicals caused malformations with only one exception (dibutylthiourea). The maximum observed frequencies of malformed embryos varied from 3 to 100% of the treated embryos.

Abnormalities, Drug-Induced↗

RIT 2214, a new biosynthetic penicillin produced by a mutant of Cephalosporium acremonium.

A number of lysine-requiring auxotrophs of Cephalosporium acremonium were investigated for incorporation of side-chain precursors and for accumulation of beta-lactam compounds. One of the auxotrophs, Acremonium chrysogenum ATCC 20389, producing cephalosporin C and penicillin N only if grown in media supplemented with DL-alpha-amino-adipic acid (DL-alpha-AAA), was found to use L-S-carboxymethylcysteine (L-CMC) as a side-chain precursor for the synthesis of a new penicillin (RIT 2214). No corresponding cephalosporin was detected. The penicillin present in the culture filtrate, was concentrated by adsorption on activated carbon and successive column chromatography on Amberlite IRA-68 and Amberlite XAD-4. Final purification was achieved by cellulose column chromatography. RIT 2214 was identified as 6-(D)-[(2-amino-2-carboxy)-ethylthio]-acetamido]-penicillanic acid by spectral analysis, bioactivity spectrum, elucidation of side-chain structure and finally by semisynthesis. Its biological properties were also evaluated.

2-Aminoadipic Acid↗

Purification, characterization, DNA sequence and cloning of a pimeloyl-CoA synthetase from Pseudomonas mendocina 35.

A pimeloyl-CoA synthetase from Pseudomonas mendocina 35 was purified and characterized, the DNA sequence determined, and the gene cloned into Escherichia coli to yield an active enzyme. The purified enzyme had a pH optimum of approximately 8.0, Km values of 0.49 mM for pimelic acid, 0.18 mM for CoA and 0.72 mM for ATP, a subunit Mr of approximately 80000 as determined by SDS/PAGE, and was found to be a tetramer by gel-filtration chromatography. The specific activity of the purified enzyme was 77.3 units/mg of protein. The enzyme was not absolutely specific for pimelic acid. The relative activity for adipic acid (C6) was 72% and for azaleic acid (C9) was 18% of that for pimelic acid (C7). The N-terminal amino acid was blocked to amino acid sequencing, but controlled proteolysis resulted in three peptide fragments for which amino acid sequences were obtained. An oligonucleotide gene probe corresponding to one of the amino acid sequences was synthesized and used to isolate the gene (pauA, pimelic acid-utilizing A) coding for pimeloyl-CoA synthetase. The pauA gene, which codes for a protein with a theoretical Mr of 74643, was then sequenced. The deduced amino acid sequence of the enzyme showed similarity to hypothetical proteins from Archaeoglobus fulgidus, Methanococcus jannaschii, Pyrococcus horikoshii, E. coli and Streptomyces coelicolor, and some limited similarity to microbial succinyl-CoA synthetases. The similarity with the protein from A. fulgidus was especially strong, thus indicating a function for this unidentified protein. The pauA gene was cloned into E. coli, where it was expressed and resulted in an active enzyme.

Acyl Coenzyme A↗

[A case of riboflavin-responsive multiple acyl-CoA dehydrogenase deficiency (glutaric aciduria type II)].

We reported a male infant with multiple acyl CoA dehydrogenase deficiency, probably due to electron transfer flavoprotein dehydrogenase deficiency. He was noted to have severe muscle weakness, a high serum creatine kinase (CK) level up to 6920 IU/L, lipid storage myopathy and fatty liver at 6 months of age. A GC/MS analysis of urinary organic acids showed excess excretion of dicarboxylic acids, including glutaric, 2-hydroxyglutaric, adipic, suberic, sebacic, malonic, ethylmalonic and methylsuccinic acids. On a urinary acylglycine analysis, hexanoylglycine and suberylglycine were increased, but not isovalerylglycine, in amount. No ketosis was noted. The muscle pathology showed increased oil-red O positive lipid droplets of various sizes indicative of lipid storage myopathy. There was diffuse decrease in the activity of cytochrome c oxidase. No ragged-red fibers were noted. His clinical symptoms improved remarkably after the administration of riboflavin (100 mg/day) and L-carnitine (1000 mg/day). He was then diagnosed as having probable riboflavin-responsive multiple acyl-CoA dehydrogenase deficiency. The glutaryl CoA dehydrogenase activity in lymphocytes was normal, as were the alpha- and beta-subunits of electron transfer flavoprotein. These findings led us to suspect electron transfer flavoprotein dehydrogenation deficiency. Although he had several episodes of short-term deterioration in clinical and laboratory findings, he developed normally with normal intelligent till 10 years of age.

Acyl-CoA Dehydrogenases↗

Adipyl crosslinked bovine hemoglobins as new models of allosteric systems.

As indicated by peptide analyses and mass spectrometry estimations, intramolecular crosslink with bis(3,5-dibromosalicyl)adipate of bovine hemoglobin results in the formation of two main components covalently bridged across the beta-cleft. In one component the crosslink joins the beta(1)V1-beta(2)K81 residues (XL-Peak-1), in the other the bridge is between the beta(1)K81-beta(2)K81 residues (XL-Peak-2). Both components are tetrameric with a mass near MW = 67 kDa as estimated by gel filtration, and a hydrodynamic radius near 3. 20 nm, estimated by dynamic light scattering. They have very low oxygen affinity with Pm near 100 mmHg (XL-Peak-1) and near 70 mmHg (XL-Peak-2) respectively at 37 degrees C, at neutral pH. The Bohr effect is almost absent in XL-Peak-1, while in XL-Peak-2 it is very near normal. Both systems show oxygen binding cooperativity with an index near n = 2.0. Flash photolysis kinetics of the recombination with CO could be resolved into a fast and a slow component. The amplitude of the fast rates were not concentration-dependent. The stopped-flow kinetics were autoaccelerating, consistent with their ligand-binding cooperativity. All rates were very similar to those of normal hemoglobin, suggesting that the oxy- rather than the deoxy-forms of the systems were affected by the crosslink. Proteins 2000;39:166-169.

Allosteric Site↗

Pharmacokinetics of sebacic acid in rats.

The pharmacokinetics of disodium sebacate (Sb) was studied in Wistar rats of both sexes. Sebacate was administered either as intra-peritoneal (i.p.) bolus (six doses ranging from 10 mg to 320 mg) or as oral bolus (two doses: 80 and 160 mg). Plasma and urinary concentrations of Sb and urinary concentrations of Sb and its products of beta-oxidation (suberic and adipic acids) were measured by an improved method using gas-liquid chromatography/mass-spectrometry. A single compartment with two linear elimination routes was selected after no increase in significance was shown by an additional compartment and after a saturable mechanism was found to be unsuitable. Both renal and non-renal elimination parameters were obtained by Marquardt non linear fitting of plasma concentrations together with urinary elimination. The data reported are calculated from the analysis on the whole population of rats and referred to an average body weight (bw) of 100 g. The Sb half-time was 31.5 min. The tissue elimination rate was 0.0122 min-1. The overall volume of distribution was found to be 26.817 ml/100 g bw. The renal clearance was 0.291 ml/min/100 g of bw, which is much less than the value of GFR reported in literature (about 1 ml/min/100 g bw), suggesting the presence of Sb reabsorption from the ultrafiltrate. The value of Sb renal clearance was found to be a concentration-independent function, suggesting the presence of a passive back-diffusion. The relative bioavailability of the oral form compared to the i.p. form was 69.09%, showing a good absorption of the drug.

Administration, Oral↗

Peroxisomes: biochemistry, molecular biology and genetic diseases- a video programme for teaching students.

Peroxisome proliferation in rodent liver is a good biochemical marker of toxicology for several classes of xenobiotics, including fibrates; phthalates and adipates; or chlorophenoxy-acetate, a herbicide. Research in peroxisomes provides a good example of the integration of fields related to basic sciences and biomedical and industrial health. A 25min video programme illustrates techniques involved in the characterization of purified peroxisomes (Fig. 1) and membranes, immunoblotting, measurement of proliferation markers, mRNA analysis at the post-transcriptional level, DNA techniques, cell cultures as biological models and computer analysis. It is aimed at undergraduates and non-biochemist advanced students in biology.

Journal Article↗

Effect of intravenous magnesium on ventricular tachyarrhythmias associated with acute myocardial infarction.

Ventricular ectopy and left ventricular dysfunction are important predictive factors for an unfavourable outcome following an acute myocardial infarction (MI). Tachyarrhythmias are a major cause of death subsequent to MI. Magnesium was postulated to have an antiarrhythmic effect after MI. Therefore we have investigated the influence of intravenous and oral magnesium (Mg) therapy on ventricular tachyarrhythmias. 67 patients with myocardial infarction (MI) diagnosed according to the WHO criteria of anamnesis, infarct-specific electrocardiogram (ECG), and enzymatic status were included in a prospective study. 23 patients (group 1) received 2 g Mg per day (= 82 mmol Mg/24 h) intravenously for the first 3 days followed by oral magnesium adipate administration of 3 x 2 coated tablets of magnesium 50 Apogepha (= 300 mg Mg/24 h or 12.34 mmol Mg/24 h, respectively) for the full duration of the study. 26 patients (group 2) received only i.v. magnesium for the first 3 days after admission (2 g Mg/24 h). The results of this treatment were compared to those of a control group of 18 MI patients without magnesium administration. All groups were identical with regard to other forms of treatment. The magnesium levels in serum and erythrocytes of all patients were measured at the following time points: days 0 (admission time), 1, 2, the day of discharge (about day 20) and after 12 weeks. The tachyarrhythmias were monitored by 24-h-continuous-electrocardiography on days 0, 1 and on the day before discharge (about day 20). The serum magnesium levels rose significantly during i.v. Mg-administration (1 and 2 day) but decreased in group 2 subsequently until the time of discharge from hospital. In contrast group 1 patients receiving oral as well as intravenous magnesium did not show this drop. The uptake of magnesium into the erythrocytes was less obvious. The erythrocyte magnesium concentration of the control group remained significantly low in serum and red blood cells. Significantly less ventricular premature beats and runs (< 5 ventricular premature beats and > 5 ventricular premature beats) compared to admission day were observed in both treated groups. These data suggest that the frequency of ventricular tachyarrhythmias is reduced by administration of intravenous magnesium and support an early high dose administration of intravenous magnesium in the wake of myocardial infarction.

Administration, Oral↗

[The development of light-curing soft denture liner].

In this study, a visible light-curing soft denture liner was developed, and 6 methacrylate polyurethane elastomer and a diluent were synthesised, their effect on tensile strength, compression set, Shore A hardness, absorption and solubility were also investigated. The results showed that the elastomer made of polyethylene adipate or polybutylene ether glycol possessed the highest tensile strength and hardness, the elastomer made of polypropylene ether glycol showed low tensile strength and low shore A hardness, the elastomer made of hydroxyl-terminated liquid polybutadiene rubber had a moderate tensile strength and hardness.

Denture Liners↗

Polyvinyl chloride-based membranes for flow injection analysis of quinine in beverages.

Potentiometric response characteristics were evaluated for quinine selective sensors based on a lipophilic ion-exchanger potassium tetrakis[3,5-bis(trifluoromethylphenyl)]borate (PTFB) immobilized together with plasticizing solvents in polyvinyl chloride membranes. The use of dioctyl phthalate (DOP), 2-nitrophenyl phenyl ether (NPPE), and bis(2-ethylhexyl)adipate (BEHA) plasticizers produced good quality quinine sensors that were sensitive and fast responding, and exhibited near Nernstian responses when used as batch-sensors. These membranes were further tested in a wall-jet flow-through potentiometric flow injection analysis (FIA) detector. Quinine sensors containing BEHA were the most suitable membrane, with no noticeable differences in sensitivity even after 5 h of continuous exposure to solutions. Interference by foreign species such as alkali, alkaline earth metal ions, sugars, and sodium benzoate was minimal in either the batch-mode (log selectivity coefficients <-2.60) or as FIA detector. When the FIA method was used, recoveries averaged 99.70% from mineral water samples spiked with 16.22 and 162.20 ppm quinine. Results for determination of quinine in other drinks such as carbonated (64.88 and 71.37 ppm) and bitter lemon drinks (55.15 ppm) agreed with those of the manufacturer's claimed values of <68, <81, and 51 ppm, respectively. With a sample throughput of 50 samples/h under optimized flow conditions, the method can be routinely used to monitor quinine in such samples.

Beverages↗

[Calcium, magnesium, phosphorus, chloride and glucose in the blood of cattle following intravenous infusion of solutions with various calcium and magnesium concentration aimed at improving the treatment of hypomagnesemia].

Recent findings on changes in the mineral content of tissues in bovine hypocalcaemia and hypomagnesaemia show the necessity of using different mineral solutions for the treatment and prevention of these conditions. Infusion of 2% CaCl2-6H2O with 10% MgCl2-6H2O and 5% glucose one after the other or in combination produced severe functional changes in cardiac activity and respiration. Recommended concentrations for treatment or prophylaxis of hypomagnesaemia were 10 g magnesium chloride and 2 g calcium chloride, or alternatively 12 g magnesium adipate and 5 g calcium gluconate, in 100 ml of distilled water (total dose 500 ml). This was well tolerated and it overcame the mineral deficiency in tissues.

Animals↗