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

A fully automated method including column-switching and isocratic high-performance liquid chromatography (HPLC) was developed for simultaneous determination of the tricyclic antidepressant clomipramine and its metabolites demethylclomipramine, 2-, 8-, and 10-hydroxyclomipramine, 2-, and 8-hydroxydemethylclomipramine and didemethylclomipramine in serum. After serum injection into the HPLC system and on-line sample clean-up on a clean-up column (Hypersil CN; 10 x 4.6 mm) by an eluent consisting of 35% acetonitrile and 65% deionized water, the chromatographic separation was performed on an analytical column (LiChrospher CN; 250 x 4.6 mm I.D.) by an eluent consisting of 38% acetonitrile and 62% aqueous sodium perchlorate (0.02 M, pH 2.5). The UV detector was set at 260 nm. The limit of quantification was about 15 ng/ml for all analytes. The coefficients of variation ranged between 3 and 12% with recovery rates between 64 and 110%. Linear regression analyses revealed coefficients of correlation between 0.98 and 0.99. The method could be applied to therapeutic drug monitoring as well as metabolism studies in man and rat.     

Carcinogenic heterocyclic amines in model systems and cooked foods: a review on formation, occurrence and intake

Frying or grilling of meat and fish products may generate low ppb levels of mutagenic/carcinogenic heterocyclic amines (HAs). Many heterocyclic amines are formed via the Maillard reaction from creatine, free amino acids and monosaccharides; compounds naturally occurring in protein-rich foods of animal origin. The formation and yield of HAs are dependent on physical parameters, such as cooking temperature and time, cooking technique and equipment, heat and mass transport, and on chemical parameters, especially the precursors to HAs. This paper reviews the current knowledge on the formation of HAs in cooked foods and model systems, and summarizes data on the content of HAs in various cooked foods, and estimates of the dietary intake of HAs. It should be noted that the presence of carcinogens of other types in food (e.g. nitrosamines, aromatic amines, cholesterol oxide products) and that their generation during frying and grilling are outside the scope of this review.     

Sparse sampling for assessment of drug exposure in toxicological studies

A dose of 200 cGy of total-body irradiation (TBI) is nonlethal in dogs: Following a granulocyte nadir in the third week post-TBI, peripheral blood cell counts recover to normal values by about 5 weeks. In the context of studies on a potential role of major histocompatibility (MHC) class II antigens in the regulation of stress hematopoiesis, we tested the effect of anti-MHC class II monoclonal antibodies (mAbs) on hematologic recovery after TBI. Thirteen dogs were given 200 cGy of TBI not followed by marrow infusion. Five received no additional treatment (concurrent controls) and eight were given daily intravenous (IV) injections of anti-class II mAbs H81.9 (anti-HLA-DR; n = 6) or B1F6 (anti-HLA-DR and -DP; n = 2) at 0.6 (n = 4) or 1.2 mg/kg/d (n = 4) on days 0-4 (n = 7) or days 0-9 (n = 1). One control dog died early from an intercurrent infection and four recovered uneventfully. Dogs given mAbs after TBI showed significantly different granulocyte and platelet kinetics. The granulocyte nadir was lower (p = 0.09) and was reached later (p = 0.005), the duration of neutropenia was longer (p = 0.08), and recovery occurred later (p = 0.02) than among controls. Similarly the platelet nadir was lower (p = 0.05), thrombocytopenia lasted longer (p = 0.02), and recovery occurred later (p = 0.02) than among controls. Four of eight mAb-treated dogs died with marrow aplasia. We propose that following irradiation, HLA-DR mediated signals result in terminal differentiation in more mature hematopoietic precursors but interfere with replication or differentiation in early hematopoietic precursors. These observations suggest a role for MHC class II molecules in the regulation of stress hematopoiesis.     

Priorities for HIV testing in developing countries?

The specificity and sensitivity of human immunodeficiency virus (HIV) antibody tests have increased and their use has expanded in developed and developing countries since 1985. Lately, more expensive rapid (US $3-12/test) and cheaper HIV tests have been proposed. HIV testing by the inexpensive enzyme-linked immunosorbent assays (ELISAs) is often infeasible in developing countries because of the lack of equipment. In 1990, in Kinshasa, Zaire, only about 50% of blood donors were screened for HIV. Counseling before and after testing at centers sponsored by non-governmental organizations is limited in developing countries. HIV testing can lead to discrimination: people have been put in quarantine because they were found to be HIV positive. In regions of high prevalence, high priority testing for blood transfusion is cost beneficial. A positive HIV test would justify the start of treatment for toxoplasmosis in a patient with a focal neurological deficit. In a region of high HIV prevalence, a patient with chronic diarrhea would benefit more from an HIV test than from gastroenterological tests. The absence of facilities for CD4 cell counting makes it impossible to advise symptom-free HIV infected subjects. Prophylactic isoniazid may decrease the incidence of Mycobacterium tuberculosis infection among HIV-infected people but the potential for non-compliance with treatment and the risk that drug resistance will develop exist. Confidential HIV testing plus counseling and condom promotion decreased the incidence of HIV infection and gonorrhea among women living in Kigali, Rwanda. There has been successful counseling of discordant couples with HIV infection in Kigali, Rwanda, and Kinshasa, Zaire. HIV testing in the most cost-beneficial way should be done for both members of a steady couple at the same time. HIV tests should be more available in developing countries, and national guidelines for HIV testing should be set.     

Bioactivation of the cooked food mutagen N-hydroxy-2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine by estrogen sulfotransferase in cultured human mammary epithelial cells

Cooked food mutagens from fried meat and fish have recently been suggested to contribute to the etiology of breast cancer. Thus, the most prevalent of these compounds, i.e. 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine, or rather its more mutagenic N-hydroxylated metabolite (N-OH-PhIP), forms DNA adducts in mammary cells, including human mammary epithelial (HME) cells. The objective of this study was to determine the involvement of estrogen sulfotransferase (EST), the only sulfotransferase identified in HME cells, in the further bioactivation of N-OH-PhIP. These studies were done in vitro using human recombinant EST and in intact HME cells. Human recombinant EST increased the covalent binding of [3H]N-OH-PhIP to calf thymus DNA approximately 3.5-fold in the presence of the sulfotransferase co-substrate 3'-phosphoadenosine-5'-phosphosulfate at each N-OH-PhIP concentration (1, 10 and 100 microM) (n = 6, P < 0.001). In contrast, EST did not catalyze the DNA binding of two other cooked food mutagens, N-hydroxy-2-amino-3-methylimidazo[4,5-f]quinoline and N-hydroxy-2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline, which are mainly hepatocarcinogens. Cultured HME cells displayed high EST activity, which could be completely inhibited by 1 microM estrone. When the cells were incubated with [3H]N-OH-PhIP, binding to native DNA occurred at 60-240 pmol/mg DNA. This binding was inhibited to 55% of control by 1 microM estrone (P < 0.01, n = 8), suggesting that EST plays a significant role in carcinogen bioactivation in human breast tissue.     

In vivo assessment of dental plaque pH changes in children after ingestion of snack foods

The aim of this study was to investigate the effects of some snack foods on plaque pH in children. In vivo dental plaque pH changes after consumption of cola, diet cola, milk, sweet biscuits, milk chocolate, banana, cheddar cheese, potato chips, boiled potato, bread and positive control of 10 percent sucrose solution were measured by using MEPH 2 pH-microelectrode. Six males and four females, ages eight to twelve years (dft: 1.3 and DMFT: 0.4) participated in this study. The measurements of plaque pH were made on forty-eight-hour accumulated plaque, at baseline to determine the resting plaque pH of the fasted plaque and at time intervals of 1, 5, 10, 15, 20, 30, 45 and 60 minutes. The plaque pH curves, delta pH values and time spent below pH 6.00 for each test product were determined. The test products were ranked according to maximum pH drop (delta pH) in ascending order as follows: cheddar cheese < diet cola < milk < banana < potato < potato chips < biscuits < cola < bread < milk chocolate < 10 percent sucrose solution.     

An assessment of the reinforcing properties of foods after amygdaloid lesions in rhesus monkeys.

Assessed the reinforcing strengths of foods in 11 rhesus monkeys before and after bilateral lesions of the lateral, basolateral, and total amygdala. Findings indicate that none of the lesions altered preoperative preferences between 3 highly palatable foods. Moreover, the lesion had no discernible effect on Ss' responses to different food rewards as measured by a progressive ratio schedule, although performance on this schedule proved sensitive to the size and type of food reward and to the degree of deprivation. Results suggest that amygdalectomy leaves a normal appreciation of at least 1 class of rewards, foods. It is concluded that the dietary changes typically seen after amygdalectomy, such as eating meat, which were also observed in the same Ss, probably reflect a loss of neophobia. (25 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Reduction of urinary mutagen excretion in rats fed garlic

Naturally occurring substances of plant origin are known to possess antimutagenic potential. Garlic (Allium sativum) was fed to rats in dried powdered form at 0.1%, 0.5% and 1% concentrations in their diet for 4 weeks. At the end of the experiment benzo[a]pyrene (1 mg/rat) was injected intraperitoneally and 24-h urine was collected from the rats. Urinary mutagens were quantitated by the Salmonella typhimurium assay. There was a significant reduction in the excretion of urinary mutagens by carcinogen-exposed rats fed garlic. Further, there was a stimulation in the activities of liver cytosolic glutathione-S-transferase and liver and lung quinone reductases. The study suggested that the antimutagenic potential of garlic may be mediated through induction of detoxification enzymes in target tissues.     

Tissue chambers--a useful model for in vivo studies of cytokine production in the pig

An in vivo tissue chamber model was developed to enable studies of local cytokine production and cellular events during inflammatory and immune reactions in the pig. Tissue chambers made of sialistic rubber tubing were surgically implanted in the subcutaneous tissue- and samples of tissue chamber fluid (TCF) and inflammatory cells were collected by aspiration with a syringe. To evaluate the model for local cytokine production, two cytokine inducers, polyribinosinic-polyribocytidylic acid (poly I:C) and fixed Aujeszky's disease virus infected PK15 cells (ADV-PK15), were injected into the tissue chambers and samples of TCF were collected 0, 4, 8, 12, 24 and 48 h post injection. Poly I:C injections induced local production of interferon-alpha (IFN-alpha) as well as tumor necrosis factor (TNF) in the TCF but kinetic differences in the production of the cytokines were noted. Poly I:C also induced an increase in cell numbers in the TCF, mainly due to increased neutrophil numbers. Injections of ADV-PK15 induced local IFN-alpha production in the TCF as long as the pigs were serologically negative to ADV. Immunofluorescence and in situ hybridization techniques could be applied for characterization of TCF cells. Moreover, cells recovered from the tissue chambers were viable and could be used in functional in vitro tests. Taken together, this tissue chamber model could prove very useful in in vivo studies of inflammatory/immune responses and cytokine production in the pig.     

Exposure assessment in industry specific retrospective occupational epidemiology studies

Quantitative estimation of exposure for occupational epidemiology studies has received increasing attention in recent years and, as a result, a body of methodological literature has begun to take form. This paper reviews the generic issues in the methodology of exposure assessment, particularly methods for quantitative retrospective assessment studies. A simple framework, termed an exposure data matrix (EDM), for defining and analysing exposure data is proposed and discussed in terms of the definition of matrix dimensions and scales. Several methods for estimation, interpolation, and extrapolation, ranging from subjective ratings to quantitative statistical modelling are presented and discussed. The various approaches to exposure assessment based on the EDM concept are illustrated with studies of lung disease among coal miners and other dust and chemically induced chronic occupational diseases. The advantages of validated statistical models are emphasised. The importance of analysis and control of errors in exposure assessments, and integration of the exposure assessment and exposure-response processes, especially for emerging occupational health issues, is emphasised.     

Targeted studies as a learning tool in outcomes assessment

BACKGROUND: Gastric lipase secretion is stimulated by gastrin in plasma, but its regulation by secretin is unknown. METHODS: In 7 normal persons we investigated the effect of exogenous secretin on the output of gastric lipase stimulated by intravenous gastrin-17. The gastric content was measured using a nasogastric tube for aspiration. The quantitative lipase secretion was measured by an enzyme-linked immunosorbant assay (ELISA) and the lipolytic activity by a kinetic assay. Plasma concentrations of secretin and gastrin were measured by radioimmunoassay. RESULTS: Gastric lipase secretion (the quantity as well as the lipolytic activity) was significantly stimulated by gastrin. In response to secretin infusion, the lipolytic activity increased as acid secretion decreased. CONCLUSION: Secretin in postprandial concentrations does not influence the quantitative gastric lipase secretion stimulated by gastrin, but it increases lipolytic activity due to inhibition of acid secretion.     

Modeling epidemiologic studies of occupational cohorts for the quantitative assessment of carcinogenic hazards

Epidemiologic studies of occupational cohorts have played a major role in the quantitative assessment of risks associated with several carcinogenic hazards and are likely to play an increasingly important role in this area. Relatively little attention has been given in either the epidemiologic or the risk assessment literature to the development of appropriate methods for modeling epidemiologic data for quantitative risk assessment (QRA). The purpose of this paper is to review currently available methods for modeling epidemiologic data for risk assessment. The focus of this paper is on methods for use with retrospective cohort mortality studies of occupational groups for estimating cancer risk, since these are the data most commonly used when epidemiologic information is used for QRA. Both empirical (e.g., Poisson regression and Cox proportionate hazards model) and biologic (e.g., two-stage models) models are considered. Analyses of a study of lung cancer among workers exposed to cadmium are used to illustrate these modeling methods. Based on this example it is demonstrated that the selection of a particular model may have a large influence on the resulting estimates of risk.     

A methodological approach for exposure assessment studies in residences using volatile organic compound-contaminated water

This paper presents a methodological approach for assessing total exposures to volatile organic compounds (VOCs) in residences using contaminated water supplies. This approach is founded on assessment of ingestion, inhalation, and dermal exposures; both long-term (i.e., 12 to 24 hr) low-level exposures and short-term (i.e., approximately 10 min) high-level exposures are considered. The methodology is based on the collection of water samples to establish the identity of the contaminants, maximum source terms, and possible dermal and ingestion exposures; integrated whole-air samples are collected to assess long- and short-term inhalation exposures; whole-air grab samples are used to confirm peak and typical inhalation exposures; and alveolar breath samples are used to confirm exposures and to estimate contaminant concentrations in the blood of the test subjects. While we do not suggest that this methodology should supersede any current investigative approach, this material is primarily offered as a consolidated reference to the many people or organizations who might contemplate a study of this type. Application of this investigative protocol should provide detailed exposure assessment information, while it supplies critical real world data for risk assessment specialists, toxicologists, and modeling experts. Data from a recent field study assessing exposures to trichloroethylene are presented to illustrate the utility and some of the limitations of this strategy.     

Epidemiologic-hygienic aspects of the assessment of carcinogenic risk in methacrylate production

The authors studied oncologic mortality among the workers exposed to some acryl monomers. The oncologic risk was estimated for the individuals exposed to acrylonitrile, methyl methacrylate, methyl- and butyl acrylate, methacrylic acid. Occurrence of the malignancies appeared to depend on the dose of methyl methacrylate.     

Preference for foods in relation to cost.

Wholesale cost of meat dishes served on the standard army installation were related to preference ratings obtained in food attitude surveys. A curve of the form[Equation omitted](Y = preference, X = cost) fitted the data closely (curvilinear r = .86). Applications of this finding are discussed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Development of a shortened center for epidemiological studies depression scale for assessment of depression in rheumatoid arthritis.

Objective: The purpose was to develop a short-form version of the Center for Epidemiologic Studies Depression Scale (CES-D) for the identification of persons with major depressive disorder (MDD) within a population with rheumatoid arthritis (RA). Study Design: Data were analyzed from 337 persons with RA. Forty-six participants met the diagnostic criteria for MDD; 291 participants were classified in the non-MDD category (including 21 participants with dysthymia and 18 participants with minor depressive disorder). A short-form version of the CES-D was developed, and multiple cutoff scores were examined. Results: A cutoff score of ≥5 from a 9-item, short-form CES-D was found to be generally as efficient as the more commonly used full-scale cutoff score of ≥16 for classifying participants with MDD within an RA population. Although the shortened CES-D scale (cutoff score ≥5) was slightly more sensitive, it also exhibited slightly less specificity than the full-scale cutoff score of ≥16. Conclusion: The results suggest that a short-form CES-D can be used to screen for MDD within an RA sample with a degree of efficiency that is generally comparable to that of the full-scale instrument. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Identification and characterization of Leuconostoc carnosum, associated with production and spoilage of vacuum-packaged, sliced, cooked ham

Leuconostoc carnosum was shown to be the specific spoilage organism in vacuum-packaged, sliced, cooked ham showing spoilage during 3 weeks of shelf life. Identification of the specific spoilage organism was done by use of phenotypic data and ClaI, EcoRI, and HindIII reference strain ribopatterns. One hundred L. carnosum isolates associated with the production and spoilage of the ham were further characterized by pulsed-field gel electrophoresis (PFGE), together with some meat-associated Leuconostoc species: L. citreum, L. gelidum, L. mesenteroides subsp. dextranicum, and L. mesenteroides subsp. mesenteroides. ApaI and SmaI digests divided the industrial L. carnosum strains into 25 different PFGE types, ApaI and SmaI types being consistent. Only one specific PFGE type was associated with the spoiled packages. This type also was detected in air and raw-meat mass samples. The spoilage strain did not produce bacteriocins. Only seven isolates belonging to three different PFGE types produced bacteriocins. Similarity analysis of the industrial L. carnosum strains revealed a homogeneous cluster which could be divided into eight subclusters consisting of strains having at most three-fragment differences. The L. carnosum cluster was clearly distinguished from the other meat-associated leuconostoc clusters, with the exception of the L. carnosum type strain. Ribotyping can be very helpful in the identification of L. carnosum, but its discriminatory power is too weak for strain characterization. PFGE provides good discrimination for studies dealing with the properties of homogeneous L. carnosum strains.