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In vivo cytogenetic effects of cooked food mutagens.

Using a variety of in vivo cytogenetic endpoints, we have investigated the effects of several compounds formed during the cooking of meat. C57Bl/6 mice were used to test for an increase in the frequency of sister-chromatid exchanges (SCEs), chromosomal aberrations, and micronucleated erythrocytes by 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx). 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (DiMeIQx), and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). MeIQx and DiMeIQx did not induce SCEs in mouse bone marrow cells. PhIP induced sister-chromatid exchanges, but not chromosomal aberrations in bone marrow. In peripheral blood lymphocytes, PhIP did induce aberrations at 100 mg/kg, the highest dose tested. PhIP induced a low but significantly increased frequency of micronuclei in normochromatic but not polychromatic erythrocytes in bone marrow and peripheral blood. However, dose responses were not observed. With the exception of the SCEs induced by PhIP, these results contrast with observations made in vitro, where these compounds were found to have significant genotoxicity in mammalian cells and a very high mutation frequency in prokaryotic systems.

Animals↗

Indoor sources of mutagenic aerosol particulate matter: smoking, cooking and incense burning.

The emission of aerosol particles and their mutagenic activity as well as the emission of some gaseous pollutants has been studied experimentally in order to compare the emission from some indoor pyrolysis processes. Cigarette (tobacco and herbal) smoking, incense and mosquito-coil burning and frying of experimental lean minced pork emitted particulate matter. Their extracts were mutagenic in the Ames Salmonella test with TA98 and activation as well as, with a higher response, in a microsuspension test with the same strain and activation condition. The response of the particles from the smoking and burning processes varied from 3000 to 50,000 revertants per gram of smoked or burnt material in the conventional Salmonella test and from 50,000 to 350,000 revertants per gram in the microsuspension assay. The frying of lean minced pork gave an airborne emission of about 53 and 560 revertants per gram of fried pork, respectively, in the 2 assays. The frying of some common food items following cookbook recipes also emitted mutagenic aerosol particles but the emitted activity was less than that in the pork experiment. Carbon monoxide, isoprene and benzene were present in the emissions from the smoking and burning processes but were not detectable in the frying fumes. The results suggest that incense and mosquito-coil burning can cause indoor air pollution akin to that from cigarette smoking. Indoor air pollution from cooking requires further study.

Aerosols↗

Carcinogenicities of heterocyclic amines in cooked food.

Mutagenic heterocyclic amines in cooked foods were carcinogenic to mice, rats and/or monkeys, when they were given orally continuously. The most common target organ was the liver, but in CDF1 mice lung tumors, forestomach tumors, lymphomas/leukemias, and blood vessel tumors in the brown adipose tissues were also induced. In F344 rats, in addition to liver tumors, tumors in the Zymbal gland, skin, clitoral gland, small and large intestines, oral cavity, and mammary gland were also induced. Monkeys given IQ developed metastatic hepatocellular carcinomas.

Amines↗

Activation of mutagens in cooked ground beef by human-liver microsomes.

The total organic base fraction purified from fried ground beef is metabolized by human-liver microsomes to form mutagens detectable by the Ames/Salmonella bacterial assay. The mutagens produced have an absolute requirement for metabolic activation; without it, no increase in the number of revertants over background is seen. Microsomes from human liver activate the mutagens significantly more than microsomes from uninduced mouse or rat liver; the microsomes from one individual were nearly as active as those of Aroclor-induced mice and rats. alpha-Naphthoflavone (ANF) inhibits activation of these mutagenic bases, implying that the metabolism is mediated by the inducible form(s) of cytochrome P-448. Thus, the human liver has the potential to metabolize the cooked beef mutagen(s) to active intermediates, posing a possible mutagenic risk. However, unlike the animal metabolizing system, which needs to be artificially induced, the human system appears to be naturally induced through diet or environmental exposure.

Animals↗

Micronucleus induction in bone-marrow cells following consumption of cooked beef in mice. Preliminary investigations.

The bone-marrow micronucleus assay was used to investigate whether the consumption of cooked meat could induce chromosome damage. There was no difference in the micronucleus frequency of mice on normal diet (cereal-based, non-purified diet) and mice on normal diet supplemented with microwaved meat. However, supplementing the normal diet with well-done pan-fried meat or rare charcoal-barbecued meat or well-done charcoal-barbecued meat produced significant increments in the micronucleus frequency of polychromatic erythrocytes--the increments were of the order of 73% (p = 0.045), 90% (p = 0.047) and 136% (p = 0.001) respectively and they were observed after a 21-day feeding trial. These results suggest that ingestion of well-done pan-fried or barbecued meat may increase genetic damage, however, the accompanying decreased intake of vegetable constituents may have also contributed to the observed changes.

Animals↗

Infectivity of Toxoplasma gondii in mutton following curing, smoking, freezing or microwave cooking.

To investigate the effects of curing with sodium chloride and sucrose, low temperature smoking, freezing at -20 degrees C, and cooking in a microwave oven, respectively, on the infectivity of Toxoplasma gondii encysted in mutton, meat from three experimentally and one naturally infected sheep was used. Samples of meat prepared accordingly as well as untreated, raw meat from each animal were assayed by mouse inoculation. Infective T. gondii was isolated from untreated samples from all animals used, but in no case from cured, smoked or frozen meat. However, in two of four steaks processed in a microwave oven, according to a standard household recipe, the parasite remained infective, possibly due to uneven heating of the meat.

Animals↗

Improvements in the microbiological quality of food samples from a hospital cook-chill system since the introduction of HACCP.

A cook-chill system of catering was introduced in Waterford Regional Hospital in December 1989. An intensive system of microbiological monitoring was put in place from day one and after 16 months a Hazard Analysis Critical Control Point (HACCP) system was introduced. Continuous monitoring showed an improvement in the microbiological quality of the food when using HACCP, with more than 90% of samples having total viable counts of less than 1 x 10(3) g-1 and the virtual elimination of pathogens. The introduction of HACCP allowed a reduction in the level of sampling and provided a useful set of records for quality assurance.

Cooking↗

Counteraction of the genotoxicity of some cooked-food mutagens by biogenic amines.

The biogenic amines tryptamine, 5-hydroxytryptamine, tyramine and histamine were assessed for their abilities to modify the genotoxicity of the cooked-food mutagens IQ, MeIQ, MeIQx, Trp-P-1 and Trp-P-2. These measurements were made using a bacterial mutation assay with hepatic fractions from either SWR mice or DSN Syrian hamsters as the activating system and Salmonella typhimurium TA98 as the indicator organism. Although histamine had very little effect on the genotoxicity of these mutagens, the other amines reduced genotoxicity, with tryptamine and 5-hydroxytryptamine exerting the greatest effect. Generally the amines exhibited greater potency when S-9 fractions from mice rather than from hamsters were used.

Animals↗

Mutagen formation during the cooking of fish.

Compounds mutagenic toward Salmonella typhimurium strains sensitive to frameshift mutation (1537, 1538 and TA98) were formed when fish flesh was fried at 190 degrees c. Four species of marine fish commonly consumed in the United States were cooked in an electric skillet and broiled beneath the elements of an electric oven. Organic extracts of the fish were tested in the Salmonella mutagenic assay using strains 1535, 1537, 1538, TA98 and TA100. Basic organic extracts of fried but not raw or broiled samples exhibited significant mutagenicity with metabolic activation. Mutagenic activity ratios ranging from 3.3 to 15.7 for the extract from 20 g of fish were observed. The mutagenicity produced during the frying of fish was dependent on time. Frying times of less than 6 min produced no mutagenic activity, while at 6 min or greater substantial mutagenicity was generated.

Animals↗

Formation of mutagens in cooked foods. IV. Effect of fat content in fried beef patties.

Cooking beef patties results in the formation of mutagens detectable by Salmonella typhimurium TA98 with metabolic activation. We now show that the amount of fat in beef affects the quantity of mutagens formed. While char increases with increasing fat, over the range of 5-309% of added fat content, mutagenicity reaches a peak at 10% added fat and subsequently decreases. Thus, char formation is not an accurate measure of mutagenicity. These results suggest that fat plays an important role in mutagen formation in fried beef.

Animals↗

History of Tropical Medicine William Ernest Cooke, FRCSI (1879-1967) and his Asian tour of 1929-1930.

Dr. W.E. Cooke was not one of the most distinguished of tropical physicians, but the specialty was heavily dependent on his likes. After several junior appointments, he was Superintendent of the Hospital for Tropical Diseases in London. He carried out an extensive tour of Asia in 1929-1930 and kept careful daily notes throughout this period, giving a valuable insight into the challenges faced by the pioneering tropical physicians and surgeons from European countries.

Asia, Southeastern↗

Development and validation of stages-of-change questions to assess consumers' readiness to use a food thermometer when cooking small cuts of meat.

Consumers' readiness to use a food thermometer when cooking small cuts of meat was assessed using Prochaska's Transtheoretical Model of Behavior Change. Face, content, and concurrent validity were assessed by peer review, cognitive interviews, and mail surveys. The self-administered mail survey was sent to two groups of Washington and Idaho residents: 1,000 randomly selected consumers (41% return rate), and 231 employees and volunteers of Cooperative Extension involved in food and nutrition education (60% return rate). Two-stage classification questions were compared with a behavior question about thermometer use, and validated using Cronbach's if-item-deleted option for alpha. Concurrent validity was assessed using cross-tabulation chi2 test. The detailed classification question more accurately classified respondents in both Consumer and Extension groups. Cronbach's alpha of the detailed question with the behavior question showed a consistency level of alpha=.73 compared to alpha=.35 for the simple question format. As expected, Consumer and Extension groups differed significantly in their stages of change (P<0.0001), verifying concurrent validity. We recommend use of the detailed classification question when staging persons related to food thermometer use. The process used for development and testing can be used to refine instruments for use in other types of interventions.

Consumer Product Safety↗

Determination of less polar heterocyclic aromatic amines in standardised beef extracts and cooked meat consumed in Austria by liquid chromatography and fluorescence detection.

An analysis method was developed for the determination of trace levels of less polar heterocyclic aromatic amines (HAs) in food samples. The development started from a frequently used sample pre-treatment scheme which was slightly improved to make it applicable with high-performance liquid chromatography (HPLC) with fluorescence detection. The method was applied for the analysis of a standardised beef extract containing 5-15 ng/g of HAs and the results are compared with those of the other participants in the same European project. In addition, the method was used for the analysis of less polar HAs in cooked meat consumed in Austria.

Amines↗

PhIP metabolites in human urine after consumption of well-cooked chicken.

We devised an assay to quantify the metabolites of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) in human urine following a single exposure to well-cooked meat. Our method uses LC/MS/MS to detect four metabolites and four deuterated internal standard peaks in a single chromatographic run. N2-OH-PhIP-N2-glucuronide was the most abundant urinary metabolite excreted by the 12 individuals who participated in our study. N2-PhIP glucuronide was the second most abundant metabolite for 8 of the 12 volunteers. The stability of PhIP metabolism over time was studied in three of the volunteers who repeated the assay eight times over a 2.5 year-period. PhIP metabolite excretion varied in each subject over time, although the rate of excretion was more constant. Our results suggest that quantifying PhIP metabolites should make future studies of individual susceptibility and dietary interventions possible.

Animals↗

An improved method for the extraction of mutagens from human urine and cooked meat using blue rayon.

A reproducible method has been developed and validated that allows the detection of mutagenic material in human urine following the intake of a meal containing pan-fried beef patties. The mutagens are extracted from the urine with blue rayon and eluted with methanol/ammonia (100:1). Using 14C-2-amino-3-methylimidazo[4,5-f]quinoline (14C-IQ) as a tracer, the extraction efficiency of heterocyclic amines was consistently found to exceed 90%. It was observed that the critical factor in attaining high extraction efficiencies was the concentration of blue rayon, which depended on the nature of the matrix. Constituents of human urine were demonstrated to compete with IQ for the binding sites on the blue rayon. For the determination of the mutagenic activity in the extracted urine, the O-acetylase over-expressing Salmonella typhimurium strain YG1024 was utilised, and the Ames test procedure was optimised with respect to S9 concentration and incorporation of a pre-incubation step. Using the developed procedure, a marked rise in mutagenic activity, determined both in the absence and presence of an activation system, was evident in human urine following the intake of a beef patty meal. Normal mutagenicity levels were restored within 24 h. Finally, a method is described, once again employing blue rayon, for the extraction of heterocyclic amines from cooked beef patties with a 92% efficiency, as exemplified by IQ.

Animals↗

Association of cooking oil fumes exposure with lung cancer: involvement of inhibitor of apoptosis proteins in cell survival and proliferation in vitro.

Cooking oil fumes (COF) have been shown to be associated with lung cancer incidence in Chinese women. Our recent report indicates that inhibitor of apoptosis protein 2 (IAP2) induced by COF may contribute to the survival and proliferation of A549 lung cancer cells. In this study, to further verify whether other antiapoptosis proteins including IAP1, X-linked IAP (XIAP), and survivin, were linked with lung cancer cell survival and proliferation, these IAPs expressions in A549 cells after treatment with COF and its two major components, benzo[a]pyrene (BaP) and 2,4-decadienal (2,4-DDE) were evaluated by Western blotting. Our data showed that IAP2 was significantly induced by COF, BaP, and 2,4-DDE, but XIAP was decreased by COF and 2,4-DDE, but not by BaP. Even though different effects of COF and 2,4-DDE on IAP2 and XIAP protein expressions were observed, the caspase-3 expression was diminished by COF and 2,4-DDE. In addition, induction of IAP2 and phosphorylated Akt proteins by COF and 2,4-DDE were simultaneously abolished by LY294002. Flow cytometry and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) analysis showed that the proportion of A549 cells at the S-phase was increased significantly after treatment with COF or 2,4-DDE. The cell proliferation induced by COF is associated with the attenuation of p21(Cip/Waf1) expression. Therefore, increases of IAP1, IAP2, survivin, and cyclin D1 expressions and decreases of XIAP, caspase-3, and p21 expressions might partly contribute to the survival and proliferation of lung cancer cells after exposure to 2,4-DDE and COF. In conclusion, the lung cancer cell growth promoted by COF might support previous epidemiological reports indicating that exposure of COF was associated with lung cancer development among Chinese women.

Air Pollution, Indoor↗

Mass spectrometric methods prove the use of beeswax and ruminant fat in late Roman cooking pots.

Lipid extracts of sherds of archaeological late Roman cooking pots were analysed using high temperature-gas chromatography coupled to a mass spectrometer and liquid chromatography with atmospheric pressure chemical ionization mass spectrometer detection (LC-APCI-MS). With these advanced techniques the use of beeswax was shown through identification of the constituting alkanes, mono and diesters. The detection of high amounts of saturated triacylglycerols (TAGs) further indicated that animal fat was processed in these pots. Part of the animal fat was characterised as originating from ruminants due to the presence of trans-fatty acids. The distribution of saturated TAGs and the higher concentration of stearic acid compared to palmitic acid in the transesterified lipid extract indicated that this was sheep fat. The results illustrate how complex mixtures can be unravelled and original contents of ancient ceramic vessels can be determined using specialised analytical equipment.

Animals↗

Analysis of nonpolar heterocyclic amines in cooked foods and meat extracts using gas chromatography-mass spectrometry.

Heat processing of muscle foods gives rise to the formation of mutagenic and carcinogenic heterocyclic amines, often at ng/g levels. A gas chromatographic-mass spectrometric (GC-MS) technique was introduced for the analysis of nonpolar heterocyclic amines in common cooked meats, pan residues, and meat extracts after solid-phase extraction. The mutagenic heterocyclic amines 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1), 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2), 2-amino-9H-pyrido[2,3-b]indole (A alpha C) and 2-amino-3-methyl-9H-pyrido[2,3-b]indole (MeA alpha C) were identified in several samples in amounts up to 8 ng/g. Also the comutagenic substances 1-methyl-9H-pyrido [3,4-b]indole (harman) and 9H-pyrido[3,4-b]indole (norharman) were detected in the samples in amounts up to almost 200 ng/g. The GC-MS method can be applied without derivatisation of the sample. The technique offers high chromatographic efficiency, yielding detection limits for pure references in the range 0.1-2 ng per injection.

Amines↗