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Fish intake and serum levels of organochlorines among Japanese women.

This study evaluates background serum levels of selected organochlorine compounds among Japanese women of reproductive age and investigates whether lifestyle factors, especially dietary factors, may be associated with these levels. A cross-sectional study was performed on 80 Japanese women, aged 26-43 years, who complained of infertility and were confirmed not to have endometriosis. The serum levels of total toxic equivalency (TEQ), 18 polychlorinated dibenzo-p-dioxins (PCDDs)/polychlorinated dibenzofurans (PCDFs), 4 coplanar polychlorinated biphenyls (cPCBs), 36 ortho-substituted polychlorinated biphenyls (PCBs), and 13 chlorinated pesticides or their metabolites were measured and data were collected on the women's age, residence, occupation, body mass index (BMI), smoking and alcohol habit and 6 dietary intakes (fish, meats, rice, vegetables, fruits and dairy products). The serum median level of total TEQ was 25.1 pg TEQ/g lipid, that of PCDDs/PCDFs/cPCBs was 11.5 pmol/g lipid, that of PCBs was 0.46 nmol/g lipid, and that of total pesticides was 1.32 nmol/g lipid. The serum levels of total TEQ, PCDDs/PCDFs/cPCBs, PCBs and pesticides were positively associated with age (P for trend=0.003, 0.01, 0.005 and 0.01, respectively) and frequent fish consumption (P for trend=0.002, 0.003, 0.0003 and 0.006, respectively). Other lifestyle factors were not associated with serum organochlorine levels. The present study suggests that Japanese women who consume fish frequently in their reproductive period tend to accumulate organochlorines in their bodies.

Adult↗

The enhancement of nonheme iron bioavailability by beef protein in the rat.

Beef protein was found to enhance the bioavailability of nonheme iron in the rat. Anemic rats were fed diets containing soy protein concentrate or rice bran as the source of nonheme iron with either lactalbumin or distilled water-washed beef, which was heme free. The criteria used to determine the relative biological value (RBV) of iron was the difference between the products of final hemoglobin x final weight and initial hemoglobin x initial weight during the repletion period. Animals fed diets with only lactalbumin as a source of dietary protein and graded levels of FeSO4 (RBV of FeSO4 = 100%) served as controls. The RBV of the endogenous iron in soy protein and rice bran was found to be 91 and 46%, respectively. Substituting washed beef for lactalbumin increased the RBV of soy protein iron to 96% (results not statistically significant) and of rice bran iron to 75% (results significant, P less than or equal to 0.05). These findings demonstrate the "meat factor" effect in the rat for the first time. Two days after completion of the 11-d hemoglobin regeneration period, the apparent absorption of iron was measured during a 60-h balance period. The apparent absorption of iron by rats fed diets containing beef tended to be higher, compared to animals fed diets containing lactalbumin.

Animals↗

Mutagens from the cooking of food. I. Improved extraction and characterization of mutagenic fractions from cooked ground beef.

Ground beef was fried at 200 degrees C (392 degrees F) to a well-done, non-charred state, and the extracted organic base fraction was found to be highly mutagenic in Salmonella strain TA1538 (6300 revertants/100 g equivalent, gE, fresh weight). The neutral and acidic extracts showed no mutagenic activity in any of the 5 standard strains of Salmonella. A new procedure based upon extraction and protein precipitation with acetone is described, which is simpler and more efficient than previously described methods. The organic base fraction was mutagenic only in Salmonella strains TA1537, TA1538, and TA98, all sensitive to frameshift mutations. Strains sensitive to base-substitution mutations showed no activity. Metabolic activation was an absolute requirement for mutagenesis; however cell toxicity was decreased by the presence of S9 activation mixture. After normal cooking, more than 20 times as much mutagenic material remained in the meat as was recovered in the pan grease and vapors. The results confirm that mutagens are formed under conventional frying conditions, and show that mutagen can be isolated by an improved extraction method.

Animals↗

Repeatability of the Petrifilm HEC test and agreement with a hydrophobic grid membrane filtration method for the enumeration of Escherichia coli O157:H7 on beef carcasses.

The Petrifilm HEC test (3M Canada Inc., London, Ontario), a quantitative microbiological test for Escherichia coli O157:H7, was evaluated for its performance as a beef-carcass monitoring test. Test repeatability and agreement and agreement with an E. coli O157:H7 detection method using a hydrophobic grid membrane filter (HGMF) overlaid onto cefixime-tellurite-sorbitol MacConkey agar (CT-SMAC) followed by a latex agglutination test for the O157 antigen were determined by using pure cultures of E. coli O157:H7, beef samples experimentally contaminated with bovine feces containing E. coli O157:H7, and naturally contaminated beef carcasses of unknown E. coli O157:H7 status from a local abattoir. The Petrifilm HEC test showed excellent repeatability and excellent agreement with the HGMF-CT-SMAC method when test samples were obtained from pure cultures and experimentally contaminated meat. All 125 naturally contaminated beef carcasses surveyed were negative for E. coli O157:H7 with both microbial methods. The Petrifilm HEC test, however, demonstrated a significantly lower proportion of cross-reactive organisms (false-positive reactions) than the HGMF-CT-SMAC method. Given the performance of this test coupled with its ease of use and compact size, it shows considerable promise for carcass testing where abattoir laboratory facilities are limited and as a substitute for more complex laboratory testing methods used in established laboratories.

Abattoirs↗

Survival of cold-stressed Campylobacter jejuni on ground chicken and chicken skin during frozen storage.

Campylobacter jejuni is prevalent in poultry, but the effect of combined refrigerated and frozen storage on its survival, conditions relevant to poultry processing and storage, has not been evaluated. Therefore, the effects of refrigeration at 4 degrees C, freezing at -20 degrees C, and a combination of refrigeration and freezing on the survival of C. jejuni in ground chicken and on chicken skin were examined. Samples were enumerated using tryptic soy agar containing sheep's blood and modified cefoperazone charcoal deoxycholate agar. Refrigerated storage alone for 3 to 7 days produced a reduction in cell counts of 0.34 to 0.81 log10 CFU/g in ground chicken and a reduction in cell counts of 0.31 to 0.63 log10 CFU/g on chicken skin. Declines were comparable for each sample type using either plating medium. Frozen storage, alone and with prerefrigeration, produced a reduction in cell counts of 0.56 to 1.57 log10 CFU/g in ground chicken and a reduction in cell counts of 1.38 to 3.39 log10 CFU/g on chicken skin over a 2-week period. The recovery of C. jejuni following freezing was similar on both plating media. The survival following frozen storage was greater in ground chicken than on chicken skin with or without prerefrigeration. Cell counts after freezing were lower on chicken skin samples that had been prerefrigerated for 7 days than in those that had been prerefrigerated for 0, 1, or 3 days. This was not observed for ground chicken samples, possibly due to their composition. C. jejuni survived storage at 4 and -20 degrees C with either sample type. This study indicates that, individually or in combination, refrigeration and freezing are not a substitute for safe handling and proper cooking of poultry.

Animals↗

Replacing protein in corn-soybean turkey diets with methionine and lysine.

The extent to which dietary protein can be minimized by using Met and Lys supplements for market turkeys has been investigated in three experiments involving 2,750 birds. Large White (Nicholas) male turkey poults were fed corn-soybean meal diets of varying protein level to supply Lys from 80 to 120 of NRC (1984) from day-old to 18 wk of age. Performance with diets of 90% NRC Lys was equal to that with diets of higher Lys, provided that SAA were at 100% of NRC. Up to .2% Lys.HCl could be substituted into the 100 or 90% of NRC diets with no depression in performance, suggesting that the other essential amino acids were present in adequate amounts. When turkeys were fed diets of 85% of NRC (1984) Lys, maximum growth and breast meat yield were obtained with supplemental SAA at 100% during 0 to 18 wk of age and Lys at 100% during 12 to 18 wk of age. The requirements for other amino acids were supplied by diets formulated to provide 85% of the Lys requirement from protein. These values were for turkeys subjected to 18 C during final growout; they were not adequate in warmer temperatures.

Animal Feed↗

Characterization and kinetics studies of water buffalo (Bubalus bubalis) myoglobin.

The colour of buffalo (Bubalus bubalis L.) meat is darker than bovine meat. Since meat colour depends on the concentration of myoglobin (Mb) and its oxidation state, we have determined the main structural and functional properties of buffalo Mb. Buffalo Mb was purified from longissimus dorsi muscles and its molecular mass determined by ESI Q-TOF mass spectrometry. The molecular mass 17,034.50 was 86.20 Da higher than the bovine Mb. This was confirmed by analysing its primary structure, using a combined approach based on Edman degradation and MALDI-TOF mass spectrometry. Comparing the amino acid sequences of both Mbs, we found three amino acid differences out of 153 amino acid residues. One is a conservative substitution (D(bov)141E(buf)), and the other two (A(bov)19T(buf) and A(bov)117D(buf)) are nonconservative. These amino acid substitutions are unlikely to cause structural changes because they are located far from the heme binding pocket, as revealed by the 3D structure of buffalo Mb elaborated by homology modelling. Stability analyses show no difference with the bovine Mb for helix E and only minor differences in the stability values for helices A and G. Moreover, autoxidation rates of purified buffalo and bovine myoglobins at 37 degrees C, pH 7.2, were almost identical, 0.052+/-0.001 h(-1) and 0.054+/-0.002 h(-1), respectively, as were their oxygen-binding Kd values, 3.7+/-0.1 microM and 3.5+/-0.1 microM, respectively. The percent of MetMb values were almost identical. The results presented here suggest that the darker buffalo meat depends on factors other than the oxidation rate of its Mb, as, for example, the Mb content (0.393+/-0.005 g/100 g of tissue) and consequently MetMb, which are almost twice as high as bovine meat (Mb: 0.209+/-0.003 g/100 g of tissue).

Amino Acid Sequence↗

Photon absorptiometric studies of elements, mixtures and substances of biomedical interest.

Photon absorptiometric measurements of several elements, mixtures and substances of biomedical interest have been performed. The 144.5 keV photons of 141Ce and the associated x-ray emissions of 203Hg at 75 keV and 137Cs at 33 keV in addition to the more commonly applied energies of 279.2 and 661.6 keV respectively, have been utilised. The concept of effective atomic numbers and the choice of tissue-equivalent substitutes has been examined with reference to the experimental findings.

Animals↗

[Diet as a source of human exposure to polychlorinated dibenzo-P-dioxins (PCDD)].

Dioxins are highly toxic by-products of many industrial processes e.g. chemical and municipal waste incineration or production of chlorophenols. These compounds penetrate the environment via air, water and soil and are then incorporated in food chains. The major source of human exposure (90% of total exposition) is consumption of a wide variety of common foods (meat, fish and dairy products) containing small amounts of dioxins. The average daily intake of PCDD via food was estimated in the range of 1-3 pg/kg bw. Common characteristics of all congeners detected in foodstuffs were 2,3,7,8-chlorine substitution. Commercial milk samples could be contaminated by PCDD/DFs present in the pulp used for the production of the milk cartoon. Also higher levels of these xenobiotics were found in samples from cows grazing in the vicinity of incinerators as compared to samples from rural farms. Human exposure to dioxin begins early in life since breast-feeding is the best way to nourish an infant. The nursing baby in developed countries get a particularly high dose of dioxins (about 85 pg TEQ/kg bw/day). Food contamination with dioxins leads to enhanced accumulation of these compounds in human tissues to the extent of exceeding acceptable level.

Diet↗

Zinc absorption from composite meals. II. Influence of the main protein source.

The absorption of zinc in man from composite meals of conventional foods with various main protein sources was measured with a radionuclide technique. The meals were extrinsically labeled with 65Zn and the absorption of zinc was determined from a measurement of the whole-body retention of the isotope about 2 weeks after ingestion of the labeled meal. The meals consisted of a meat patty made from either chicken, beef, or soybeans, and included potatoes, tomatoes, and white bread. For meals based on animal protein a positive correlation was found between zinc absorption and zinc content. When 25% of the animal protein in the chicken meal was replaced by defatted soy flour neither the zinc content nor zinc absorption were significantly influenced. The same substitution in a beef meal resulted in a decrease in zinc content and also a lower zinc absorption. The absorption from a soybean meal did not differ from an animal protein meal with the same zinc content. Lower zinc absorption was found when the calcium content of a soybean meal was increased by the addition of milk. For this study, the total zinc content of the meal was the most important factor influencing the amount of zinc absorbed.

Adult↗

Genetic characterization and expression of the novel fungal protease, EPg222 active in dry-cured meat products.

EPg222 protease is a novel extracellular enzyme produced by Penicillium chrysogenum (Pg222) isolated from dry-cured hams that has the potential for use over a broad range of applications in industries that produce dry-cured meat products. The gene encoding EPg222 protease has been identified. Peptide sequences of EPg222 were obtained by de novo sequencing of tryptic peptides using mass spectrometry. The corresponding gene was amplified by PCR using degenerated primers based on a combination of conserved serine protease-encoding sequences and reverse translation of the peptide sequences. EPg222 is encoded as a gene of 1,361 bp interrupted by two introns. The deduced amino acid sequence indicated that the enzyme is synthesized as a preproenzyme with a putative signal sequence of 19 amino acids (aa), a prosequence of 96 aa and a mature protein of 283 aa. A cDNA encoding EPg222 has been cloned and expressed as a functionally active enzyme in Pichia pastoris. The recombinant enzyme exhibits similar activities to the native enzyme against a wide range of protein substrates including muscle myofibrillar protein. The mature sequence contains conserved aa residues characteristic of those forming the catalytic triad of serine proteases (Asp42, His76 and Ser228) but notably the food enzyme exhibits specific aa substitutions in the immunoglobulin-E recognition regions that have been identified in protein homologues that are allergenic.

Amino Acid Sequence↗

hOGG1 Ser326Cys polymorphism modifies the significance of the environmental risk factor for colon cancer.

AIM: To determine the association of hOGG1 (8-oxoguanine glycosylase I, OGG1) polymorphism of Ser326Cys substitution with colon cancer risk and possible interaction with known environmental risk factors. METHODS: A case-control study with 125 colon cancer cases and 247 controls was conducted. RESULTS: There was no major difference in Ser326Cys genotype distribution between cases and controls. The meat intake tended to increase the odds ratio for colon cancer with an OR of 1.72 (95 % confidence interval; CI=1.12-2.76). Such tendency was more prominent in Cys/Cys carriers (OR=4.31, 95 % CI=1.64-11.48), but meat intake was not a significant risk factor for colon cancer in Ser/Ser or Ser/Cys carriers. The OR for colon cancer was elevated with marginal significance in smokers who were Cys/Cys carriers (OR=2.75, 95 % CI=1.07-7.53) but not in Ser/Ser or Ser/Cys carriers. CONCLUSION: These results suggest that the hOGG1 Ser326Cys polymorphism is probably not a major contributor to individual colon cancer susceptibility overall, but the Cys/Cys genotype may alter the impact of some environmental factors on colon cancer development.

Base Sequence↗

Hypocholesterolemic effect of substituting soybean protein for animal protein in the diet of healthy young women.

The effect of dietary protein on the level of plasma cholesterol in young, healthy, normolipidemic women was investigated in two separate studies by feeding either a conventional diet containing mixed protein, or a plant protein diet in which the animal protein of the first diet was replaced by soy protein meat analogues and soy milk. The diets were similar with respect to carbohydrate, fat and sterol composition. The first study, lasting 73 days and involving six subjects, gave an indication that plasma cholesterol levels were lower on the plant protein diet. The second study, which incorporated a number of improvements based on experience, lasted 78 days and used a cross-over design involving two groups of five subjects each. In this study, the mean plasma cholesterol level was found to be significantly lower on the plant protein diet.

Adult↗

Heat treatment modifies the allergenicity of beef and bovine serum albumin.

The effect of heat on the allergenicity of beef and bovine serum albumin was investigated among 10 toddlers skin prick test (SPT)-positive to raw and cooked beef. The meat-allergy diagnosis was confirmed during double-blind, placebo-controlled food challenge (DBPCFC) with 180 g of beef cooked for 5 min at 100 degrees C. SPT with homogenized and freeze-dried beef, and heated and unheated bovine serum albumin were performed. Both heated and unheated bovine serum albumin, homogenized beef, and freeze-dried beef were used in trial DBPCFC. All children were SPT-positive to unheated bovine serum albumin. Seven were positive to heated bovine serum albumin, one to freeze-dried beef, and none to homogenized beef. DBPCFCs were negative for homogenized beef and freeze-dried beef, positive for unheated bovine serum albumin in five patients, and positive for heated albumin in four children. We conclude that heating reduces sensitization to beef and bovine serum albumin but does not abolish reactivity to albumin under home conditions. However, industrially heat-treated and sterilized homogenized beef and freeze-dried beef may be suitable substitutes in beef-allergic children's diets.

Allergens↗

Quantitative structure-activity relationships of heterocyclic amine mutagens formed during the cooking of food.

The major protein-rich foods, particularly muscle meats, contain part-per-billion quantities of potent mutagens formed by frying or broiling to a well-done state. Related mutagens are formed by pyrolysis of amino acids or proteins and in heated model systems. The thermic mutagens so far identified are heterocyclic aromatic amines of aminoimidazo-azaarene (AIA) and aminocarboline classes. The chemicals require activation by enzymes to form metabolites reactive with nucleic acids. These thermic mutagens, and numerous synthetic congeners, exhibit an enormous range of potency as frameshift mutagens in the Ames/Salmonella assay. However, structural variations are nominal within the two classes. Structural parameters that appeared relevant to determining potency were selected for 38 AIAs and 23 amino-carbolines. For the AIA class these were: the number of fused rings, the number of heteroatoms in Rings 2 and 3, methyl substitution on imidazo ring nitrogen atoms, and methyl substitution on ring carbon atoms. For the amino-carboline class the structural parameters were: the position of the pyridine-type nitrogen atom in Ring 1, the substitution position of the exocyclic amino group on Ring 1, and methyl substitution on ring carbon atoms. These structural parameters may influence mutagenic potency in the following ways. 1) Electronic or steric effects may determine the reactivity and stability of the ultimate mutagenic metabolite. Optimal balance of reactivity and lifetime of this transient intermediate may be required for access to and reaction with nuclear DNA to cause mutations. 2) Substitution on the rings may block detoxication reactions. The structural parameters identified should prove useful in predicting the mutagenicity of untested compounds of these types.

Amines↗

DNA adduct levels and intestinal lesions in congenic rapid and slow acetylator syrian hamsters admi food mutagens 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) or 2-amino-3-methylimidazo[4,5-f]quinoline (IQ).

Epidemiological studies indicate that rapid acetylators with a high intake of well-done red meat have an increased risk of colorectal cancer. Arylamine N-acetyltransferase enzymes (E.C. 2.3.1.5) activate carcinogenic heterocyclic amines found in the crust of fried meat via O-acetylation of their N-hydroxylamines to reactive intermediates that bind covalently to DNA and produce mutations. Syrian hamsters as well as humans express two N-acetyltransferase isozymes (NAT1 and NAT2) which differ in substrate specificity and genetic control. Nucleic acid substitutions in the NAT2 gene segregate individuals into rapid, intermediate and slow acetylator phenotypes. In the present paper, we examined the role of the polymorphic NAT2 acetylator genotype in carcinogenesis induced by the food mutagens 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) or 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) by comparing Syrian hamster lines congenic at the NAT2 locus. No differences were found between rapid and slow acetylator congenic hamsters in levels of intestinal PhIP-DNA adducts. In contrast to previous studies in rats, no carcinogen-related induction of the preneoplastic lesions aberrant crypt foci or tumors was found in the intestines of rapid and slow acetylator congenic Syrian hamsters administered PhIP or IQ.

Acetylation↗

[Use of earthworms as a protein supplement in diets for rabbits].

The purpose of the present study was to evaluate the nutritive value of earthworms as protein feed in rabbit rations. Earthworm meal was obtained from Eisenia foetida and Lumbricus rubellus. Its proximate chemical composition, amino acid composition and protein digestibility in vitro were determined. In addition, growing rabbits were fed a diet containing 30% of the total protein as earthworm meal, diet which was compared with a control diet containing soybean meal as protein feed. Both diets were isocaloric and isonitrogenous. Feed intake, weight gain, feed conversion and apparent digestibility were measured. Results showed high protein (50.86%) and fat (10.16%) contents, and low fiber percentage (2.67%). Amino acids content including the essential, and in vitro protein digestibility percentage were similar to fish and meat meals and higher than soybean meal. There were no differences in feed intake, weight gain and feed conversion. Apparent digestibility was 5.09% higher (P less than 0.05) with the diet containing earthworm meal than with the control diet. It was concluded that it is possible to substitute 30% of the protein in the diet of growing rabbits, with earthworm meal, without any adverse physiological effects. Similar results to those achieved when conventional protein supplements are used for rabbit rations, were obtained.

Amino Acids↗