Characterizing the pre-clinical phase of inflammatory bowel disease.

Understanding the biological changes that precede a diagnosis of inflammatory bowel disease (IBD) could facilitate pre-emptive interventions, including risk factor modification, but this pre-clinical phase of disease remains poorly characterized. Using measurements from 17 hematological and biochemical parameters taken up to 10 years before diagnosis in over 20,000 IBD patients and population controls, we address this at massive scale. We observe widespread significant changes in multiple biochemical and hematological parameters that occur up to 8 years before diagnosis of Crohn's disease (CD) and up to 3 years before diagnosis of ulcerative colitis. These changes far exceed previous expectations regarding the length of this pre-diagnostic phase, revealing an opportunity for earlier intervention, especially in CD. In summary, using a nationwide, case-control dataset-obtained from the Danish registers-we provide a comprehensive characterization of the hematological and biochemical changes that occur in the pre-clinical phase of IBD.

Medication-Wide Study: Exploring Medication Use 10 Years Before a Diagnosis of Inflammatory Bowel Disease.

INTRODUCTION: There is growing interest in the prediagnostic phase of inflammatory bowel disease (IBD) and in the overlap of IBD with other diseases. We described and compared use of any prescription medication between individuals with and without IBD in a 10-year period preceding diagnosis. METHODS: Based on cross-linked nationwide registers, we identified 29,219 individuals diagnosed with IBD in Denmark between 2005 and 2018 and matched to 292,190 IBD-free individuals. The primary outcome was use of any prescription medication in years 1-10 before IBD diagnosis/matching date. Participants were considered as medication users if they redeemed ≥1 prescription for any medication in the World Health Organization Anatomical Therapeutic Chemical (ATC) main groups or subgroups before diagnosis/matching. RESULTS: The IBD population had a universally increased use of medications compared with the matched population before IBD diagnosis. At 10 years before diagnosis, the proportion of users was 1.1-fold to 1.8-fold higher in the IBD population in 12 of 14 ATC main groups of medication ( P -value < 0.0001). This applied across age, sex, and IBD subtypes, although it was the most pronounced for Crohn's disease (CD). Two years before diagnosis, the IBD population had a steep increase in medication use for several organ systems. When analyzing therapeutic subgroups of medication, the CD population exhibited 2.7, 2.3, 1.9, and 1.9 times more users of immunosuppressants, antianemic preparations, analgesics, and psycholeptics, respectively, than the matched population 10 years before diagnosis ( P -value < 0.0001). DISCUSSION: Our findings demonstrate universally increased medication use years before IBD, especially CD, diagnosis and indicates multiorgan involvement in IBD.

Endocrine disease history and the risk of postpartum depression.

BACKGROUND: Previous research has suggested that some women are at increased risk of postpartum depression (PPD) because of an extra sensitivity to fluctuating hormones before and after parturition. This may particularly apply to women with endocrine disease, characterised by a less than optimal capability to self-regulate the hormonal feedback system. AIMS: To investigate if women with endocrine disease history are at increased risk of developing PPD. METHOD: Based on information from Danish national registers, this nationwide cohort study included 888 989 deliveries (1995-2018). Endocrine disease history was defined as thyroid disease, pre-pregnancy diabetes, polycystic ovary syndrome and/or previous gestational diabetes within 10 years before pregnancy start. PPD was defined as use of antidepressants and/or hospital contact for depression within 6 months after childbirth. RESULTS: Among 888 989 deliveries, 4.1% had a history of endocrine disease and 0.5% had a PPD episode. Overall, women with an endocrine disease history had a 42% (risk ratio 1.42, 95% CI 1.24-1.62) higher risk of PPD when compared with women with no endocrine disease. However, we also found the reverse association, whereby women with a PPD history had a 50% (hazard ratio 1.5, 95% CI 1.4-1.6) higher risk of endocrine disease when compared with women with no PPD history. CONCLUSIONS: Women with endocrine disease history had a 40% higher risk of PPD compared with women with no endocrine disease. More attention should be given to pregnant women with endocrine disease history to increase awareness of early signs of PPD. The bi-directionality of the association points to a common underlying factor.

Familial risk of postpartum depression.

OBJECTIVE: Many psychiatric diseases have a strong familial aggregation, but it is unknown whether postpartum depression (PPD) without prior psychiatric history aggregates in families. METHODS: Based on Danish national registers, we constructed a cohort with information on 848,544 singleton deliveries (1996-2017). Women with an episode of PPD were defined as having used antidepressant medication and/or had a hospital contact for depression within 6 months after delivery. Those with psychiatric history prior to the delivery were excluded. We estimated relative risk (RR) of PPD, comparing women with female relatives with and without PPD history, respectively. RESULTS: Overall, women with a PPD history in female blood relatives had themselves a higher risk of PPD (RR = 1.64, 95% CI 1.16-2.34). Having the first-degree female relative with PPD history was associated with a more than 2.5 times (RR = 2.65, 95% CI 1.79-3.91) increased risk of PPD. However, having the second/third-degree female relative and/or a female non-blood relative with PPD history did not increase the woman's own risk of PPD (RR = 0.58, 95% CI 0.26-1.28, RR = 1.09, 95% CI 0.83-1.44). CONCLUSION: Postpartum depression aggregates in families with no other psychiatric history, but the findings do not support a strong genetic trait as a major cause. Other possible mechanisms are shared environment and/or health-seeking behavior in close relationships.

Psychometric properties and reference data for Danish versions of Free and Cued Selective Reminding Test, Category Cued Memory Test and Logical Memory.

Memory assessment is a key element in neuropsychological testing. Gold standard evaluation is based on updated normative data, but in many small countries (e.g. in Scandinavia) such data are sparse. In Denmark, reference data exist for non-verbal memory tests and list-learning tests but there is no normative data for memory tests which capture narrative recall and cued recall. In a nation-wide study, Free and Cued Selective Reminding Test (FCSRT), WMS-III Logical Memory (LM) and a newly developed test Category Cued Memory Test (CCMT-48) were applied in 131 cognitively intact persons (aged 60-96 years). Regression-based reference data for Danish versions of FCSRT, CCMT-48 and LM adjusted for age, education and gender are provided. Gender and age-group had a significant impact on the expected scores, whereas the effect of education had a limited effect on expected scores. Test performances were significantly correlated in the range 0.21-0.51. Based on these findings and previous results it may be relevant to assess both free recall, cued recall and recognition to tap the earliest changes associated with neurodegeneration, and this study therefore provides an important supplement to existing Danish normative data. Future studies should investigate the discriminative validity of the tests and the clinical utility of the presented reference data.

EuroFIR eBASIS: application for health claims submissions and evaluations.

BACKGROUND: The European Food Information Resource (EuroFIR) network has established the eBASIS (Bioactive Substances in Food Information System) online food composition and biological effects database for plant-derived bioactive compounds (phytochemicals). On the basis of submitted evidence, the European Food Safety Authority (EFSA) expert panel on Dietetic Products, Nutrition and Allergies assesses whether claims made under articles 13.1, 13.5 or 14 of the Regulation (EC) 1924/2006, which governs the use of nutrition and health claims on foods, are scientifically justified. This report evaluates the eBASIS biological effects database in the preparation and evaluation of health claims dossiers. METHODS: The eBASIS biological effects database is a compilation of expert-evaluated data extracted from the literature, prioritizing human intervention studies to investigate health effects of phytochemicals. Currently included are >750 records from 445 studies providing data on 56 validated biomarkers, mainly relating to cardio-metabolic and bone health outcomes. The data cover 144 bioactive compounds from 17 compound classes. Using the EFSA Register of Questions and the database of general function health claims, we identified claims relating to phytochemicals made under articles 13.1, 13.5 and 14 and compared them with the eBASIS database to identify overlap between them. RESULTS: The EFSA online health claims database contains 4240 submissions under article 13.1, of which 2157 pertain to plants or plant-based bioactive compounds; 496 of these relate to plants or bioactive compounds included in the eBASIS biological effects database. Out of the 18 current 13.5 'new function' claims on EFSA's register of questions, 7 are for plants or plant-based bioactive compounds, of which 6 are included in eBASIS. Of the 222 defined article 14 claims, 21 pertain to plants or plant-based bioactive compounds, of which 19 are in eBASIS. CONCLUSIONS: There is extensive overlap between eBASIS and the submitted health claims that relate to plant-based bioactive compounds. EuroFIR eBASIS is a useful tool for regulators to independently check completeness of health claims applications relating to phytochemicals and is a potentially valuable resource to assist claimants in the compilation of dossiers on functional foods and health claims.

Information on plant foods in eBASIS: what is in a correct botanical scientific name?

This paper presents the plant information included in the eBASIS (BioActive Substances in Foods Information System) database on composition and biological activity of selected bioactive compounds from European plant/mushroom foods with putative beneficial and/or toxic effects. The European Food Information Resource (EuroFIR)-NETTOX Plant List (2007) presents scientific and vernacular names in 15 European languages for around 325 major European plant/mushroom foods and also for different parts of these foods. This list and its predecessor, the NETTOX List of Food Plants, have been used by national food authorities and within the European Union for consideration of plants and mushrooms that have been used to a significant degree up to 1997 and are therefore not covered by the novel food regulation (European Parliament and Council of the European Union, 1997). The species and the plant part studied are insufficiently characterised in many scientific papers. This paper informs about the naming of plants and mushrooms as an aid for scientists who are not botanists or mycologists themselves. Knowledge on scientific names used, including synonyms, may also be important for finding all relevant papers when searching the literature. In many cases, vernacular/trivial names in, for example, English do not uniquely identify the species. Finally, recommendations are given to assist researchers and reviewers of papers dealing with botanical/mycological information.

Agaritine content of 53 Agaricus species collected from nature.

Fifty-three different species of the genus Agaricus were collected in the Czech Republic during the period 1998-2001 and identified by an experienced mycologist. The samples were analysed for agaritine (N2-(gamma-L-glutamyl)-4-hydroxymethylphenylhydrazine) content, a precursor to a suspected rodent carcinogen. There was a huge variation in agaritine content between species, but less variation between samples of a species. Whereas the cultivated mushroom Agaricus bisporus commonly contain 200-500 mg agaritine kg(-1) fresh weight, no less than 24 of the 53 species contained agaritine levels above 1000 mg kg(-1) fresh weight. The highest level was found in A. elvensis containing up to 10, 000 mg kg(-1) fresh weight. Twenty species contained intermediate levels (100-1000 mg kg(-1)), and nine species were below 100 mg kg(-1). Some of the species producing low levels of agaritine might be candidates for future strain development of Agaricus mushrooms for cultivation. No correlation could be observed between agaritine content and size of the mushroom, week of the year when collected, year of collection, or site of collection. Besides occurring in the genus Agaricus, some species of the genera Leucoagaricus and Macrolepiota were also shown to contain agaritine.

Subchronic oral toxicity study on the three flavouring substances: octan-3-ol, 2-methylcrotonic acid and oct-3-yl 2-methylcrotonate in Wistar rats.

Groups of 10 male and 10 female rats were administered 0, 25, 100 or 400 mg octan-3-ol/kg body weight per day, 77 mg 2-methylcrotonic acid/kg body weight per day or 163 mg oct-3-yl 2-methylcrotonate/kg body weight per day by gavage for 90 days. Relative liver weights of high-dose octan-3-ol males, and males and females dosed with oct-3-yl 2-methylcrotonate were significantly greater than those of the control. In male and female rats dosed with the highest level of octan-3-ol and in male rats dosed with 2-methylcrotonic acid, incidences of bile duct proliferation were increased. In the kidneys of males dosed with mid- and high level of octan-3-ol and oct-3-yl 2-methylcrotonate, tubular karyomegaly and desquamation of tubular epithelial cells were observed. Based on increased liver weight and microscopic evaluation of the liver and kidney, a no-observed-effect level (NOEL) of 25 mg/kg for octan-3-ol in rats was established. The histopathological evaluation of the liver of rats dosed with oct-3-yl 2-methylcrotonate revealed lesions corresponding to the lesions seen in rats dosed mid-dose with octan-3-ol. This observation is in accordance with the general assumption that oct-3-yl 2-methylcrotonate is completely hydrolysed to octan-3-ol and 2-methylcrotonic acid. However, when comparing the liver histopathology of oct-3-yl 2-methylcrotonate and 2-methylcrotonic acid and the kidney lesions of all three substances, conflicting results were seen and the present study does not allow the conclusion to be drawn that oct-3-yl 2-methylcrotonate and structurally-related esters are completely hydrolysed, at least under the conditions of the present study.

Stability of agaritine - a natural toxicant of Agaricus mushrooms.

Agaritine (N-(gamma-L(+)-glutamyl)-4-hydroxymethylphenylhydrazine) is a phenylhydrazine derivative found in the cultivated Agaricus mushroom which is claimed to give rise to carcinogenic products when metabolized. The stability of a synthetic sample of agaritine was tested in water and methanol. In tap water kept in open vials, agaritine was totally degraded within 48h. Since agaritine degradation was less pronounced in closed than in open vials, and slower in Milli Q water and, in particular, in Milli Q water purged with N(2), the degradation seems to be oxygen-dependent. The antioxidant dithiothreitol reduced the degradation. Four or possibly five ultraviolet-absorbing compounds were formed during degradation, but these have not yet been identified. Whereas the rate of degradation was similar at temperatures between 4 and 22 degrees C, it was quicker at an acidic than at a neutral pH. The latter observation was confirmed in experiments where agaritine was incubated in simulated gastric fluid (pH 1.2). The importance of the degradation when performing toxicological studies with agaritine is discussed.

Influence of storage and household processing on the agaritine content of the cultivated Agaricus mushroom.

Agaritine (N-(gamma-L(+)-glutamyl)-4-hydroxymethyl-phenylhydrazine) was identified and quantified by high-pressure liquid chromatography and used as a marker for the occurrence of phenylhydrazine derivatives in the cultivated Agaricus bitorquis and A. garicus hortensis mushrooms. Although relatively high levels of agaritine (around 700 mg kg(-1)) could be found in freshly harvested A. bitorquis from early flushes, samples from supermarkets contained less agaritine. The content of 28 samples varied between 165 and 457 mg kg(-1), on average being 272 +/- 69 mg kg(-1). The highest amounts of agaritine were found in the skin of the cap and in the gills, the lowest being in the stem. There was no significant difference in agaritine content of the two mushroom species in our study. Pronounced reduction in agaritine content was observed during storage of mushrooms in the refrigerator or freezer, as well as during drying of the mushrooms. The degree of reduction was dependent on the length and condition of storage and was usually in the region 20-75%. No reduction in agaritine content was observed during freeze-drying. Depending on the cooking procedure, household processing of cultivated Agaricus mushrooms reduced the agaritine content to various degrees. Boiling extracted around 50% of the agaritine content into the cooking broth within 5min and degraded 20-25% of the original agaritine content of the mushrooms. Prolonged boiling, as when preparing a sauce, reduced the content in the solid mushroom further (around 10% left after 2h). Dry baking of the cultivated mushroom, a process similar to pizza baking, reduced the agaritine content by approximately 25%, whereas frying in oil or butter or deep frying resulted in a more marked reduction (35-70%). Microwave processing of the cultivated mushrooms reduced the agaritine content to one-third of the original level. Thus, the exposure to agaritine was substantially less when consuming processed Agaricus mushrooms as compared with consuming the raw mushrooms. However, it is not yet known to what extent agaritine and other phenylhydrazine derivatives occurring in the cultivated mushroom are degraded into other biologically active compounds during the cooking procedure.

Agaritine content in processed foods containing the cultivated mushroom (Agaricus bisporus) on the Nordic and the Czech market.

The level of agaritine was measured in fresh and canned cultivated mushroom (Agaricus bisporus) as well as in other food products containing A. bisporus, by reversed phase high performance liquid chromatography. The two fresh samples were purchased on the open market and contained 212 and 229 mg/kg, respectively. Of the 35 different trademarks of canned mushroom products studied, 25 were based on cut mushrooms and 10 on whole mushrooms. On average, whole mushrooms contained 14.9 +/- 6.7 mg agaritine per kg product whereas cut mushrooms contained 18.1 +/- 7.8 mg/kg. There was no statistically significant difference between these two values. Agaritine levels in brine were generally slightly lower than the levels detected in canned mushrooms. Thus, the level of agaritine in A. bisporus is reduced more than 10 times during the wet canning process, resulting in low levels in canned products. On a portion basis, somewhat higher amounts of agaritine may be found in some other food products (mushroom soup and pasta sauce) containing A. bisporus.

Food plant toxicants and safety Risk assessment and regulation of inherent toxicants in plant foods.

The ADI as a tool for risk management and regulation of food additives and pesticide residues is not readily applicable to inherent food plant toxicants: The margin between actual intake and potentially toxic levels is often small; application of the default uncertainty factors used to derive ADI values, particularly when extrapolating from animal data, would prohibit the utilisation of the food, which may have an overall beneficial health effect. Levels of inherent toxicants are difficult to control; their complete removal is not always wanted, due to their function for the plant or for human health. The health impact of the inherent toxicant is often modified by factors in the food, e.g. the bioavailability from the matrix and interaction with other inherent constituents. Risk-benefit analysis should be made for different consumption scenarios, without the use of uncertainty factors. Crucial in this approach is analysis of the toxicity of the whole foodstuff. The relationship between the whole foodstuff and the pure toxicant is expressed in the `product correction factor' (PCF). Investigations in humans are essential so that biomarkers of exposure and for effect can be used to analyse the difference between animals and humans and between the food and the pure toxicant. A grid of the variables characterising toxicity is proposed, showing their inter-relationships. A flow diagram for risk estimate is provided, using both toxicological and epidemiological studies.

Failure of the cultivated mushroom (Agaricus bisporus) to induce tumors in the A/J mouse lung tumor model.

We studied whether the cultivated mushroom (Agaricus bisporus) or 4-(carboxy)phenylhydrazine (CP) induce lung adenomas in the A/J mouse lung tumor model. For 26 weeks female mice were fed a semisynthetic diet where 11 or 22% of the diet was replaced by freeze-dried mushrooms. The intake of the mushroom diets was equivalent to an intake of agaritine, the major phenylhydrazine derivative occurring in the mushroom, of 92 or 166 mg/kg body weight per day. The intake of CP was 106 mg/kg body weight per day. Neither the freeze-dried mushroom nor CP induced statistically significant increased numbers of lung adenomas in female A/J mice in the administered dosages.

Carcinogenicity study of the emulsifier TOSOM and the release agent TOS in Wistar rats.

Groups of 60 Wistar rats of each sex were fed diets containing 3, 6 or 12% of the margarine emulsifier TOSOM (thermally oxidized soybean oil interacted with mono- and diglycerides of fatty acids) for 2.5 yr. In addition, three groups of 60 rats of each sex were fed two products of the release agent TOS (thermally oxidized soybean oil) in dietary levels of 1.2% TOS(G) (TOS from Grindsted Product A/S, Denmark) and 0.3 and 1.2% TOS(N) (TOS from Nexus Aps, Denmark), respectively for 2.5 yr. 120 rats of each sex fed a diet containing mono- and diglycerides served as controls. The diets given to all groups were isocaloric. Clinical appearance, food consumption, body weight and weight gain, survival, haematology, and clinical chemistry parameters were examined. Gross and histopathological examinations, including neoplastic and non-neoplastic lesions, were performed on all groups. Time to occurrence of tumours was recorded. No substance-related effect, including carcinogenicity, was found.

HPLC profiles of mutagens in lean ground pork fried at different temperatures.

Ground lean pork was formed into patties and fried under ordinary conditions making sure that the crust was not charred. No fat was added when frying. The meat was fried at pan temperatures of 200 degrees C, 250 degrees C, and 300 degrees C until the temperature at the centre of the patties was either 65 degrees C or 70 degrees C. The crust was extracted with aqueous acid followed by concentration of the mutagens on an XAD-2 column and elution with acetone. The total mutagenic activity and high-pressure liquid chromatography (HPLC) analysis of the mutagenic components ("mutagrams") in the eluates were determined for the different frying procedures using the Salmonella/mammalian microsome test strain TA 98. Each 50 degrees C increase in the pan temperature (from 200 degrees C to 250 degrees C and from 250 degrees C to 300 degrees C) resulted in a doubling of the total mutagenic activity. The HPLC profiles of the mutagens were quite similar for the different frying temperatures, although a strong increase in the relative amount of more apolar mutagens was seen at 300 degrees C (the highest temperature). The major mutagenic activity of the HPLC fractions was confined to seven regions (mutagenic peaks) and a comparison of the HPLC profiles of the mutagens in fried beef and pork patties showed identical profiles. It is therefore concluded that the mutagenic compounds formed in fried beef and pork are similar in structure.

Animal feeding study with nitrite-treated meat.

In order to detect possible formation of carcinogenic N-nitroso compounds from nitrite and nitrosatable compounds in meat, studies were carried out with 70 male and 140 female F0 rats, divided into six groups, and 60, 100, 70, 60, 60 and 66 of their male and female offspring. One control group received casein and other groups chopped pork as the sole protein source (45%, mass/mass) on a fresh basis, either salted (sodium chloride) or not. For test groups, nitrite was also added to the meat before autoclaving and storing the diet and represented mass fractions of 200, 1 000 and 4 000 mg/kg, as sodium nitrite. The results do not demonstrate any effect on reproduction and no significant carcinogenic effect was revealed. However, an observed tendency toward an increased number of tumour-bearing rats in the highest dose group, plus the possible formation of carcinogenic N-nitroso compounds in nitrite-treated meat products, led to a recommendation to reduce the use of nitrite. Results from a concomitant study demonstrate that it is possible to produce many cured-meat products with the addition of only 50 mg/kg nitrite.

Effects of rapeseed oil compared to hydrogenated marine oil in rats. Histopathological effects of erucic acid and isomers on the heart.

In a recent review on the role of fats in human nutrition Vergroesen and Gottenbos (1975) emphasize, that the presence of erucic acid isomers in hydrogenated marine oils (HMO) may constitute a toxicological problem as serious as erucic acid in rapeseed oil (RSO). On the contrary Christophersen et al. (1976) stress, that these fatty acids in HMO represent a minor toxicological problem, and that conclusions drawn from HMO to RSO are questionable. Preliminary studies at our institute indicate lipidosis in the heart both in rats fed RSO and HMO, containing equivalent amounts of erucic acid and erucic acid isomers respectively. Further experiments of longer duration are performed with HMO and RSO in order to compare the effect on the heart, especially in relation to the formation of cell necrosis.

Metabolism of L(+)-and D(-)-tartaric acids in different animal species.

L(+)-Tartaric acid, which is the naturally occurring form, is used as a food additive in a variety of foodstuffs. Recently, there has been an increasing interest in using synthetic (D,L)-tartaric acid as a substitute. Several toxicological studies have been published on L(+)-tartaric acid, but practically nothing concerning the racemic form. The metabolism of L(+)-tartaric acid has been investigated in a variety of species including man. Species differences have been noted in the ability to excrete the acid in the urine after oral administration. The decomposition of tartaric acid by the intestinal flora has been implicated as an important factor. (Underhill et al., 1931 a,b). In the present study the metabolic fate of L(+)- and D(-)-tartaric acids is compared in different species, in vivo after oral administration and in vitro after incubation with caecal extract.