Ulrich Halekoh

Multielemental Fingerprinting as a Tool for Authentication of Organic Wheat, Barley, Faba Bean, and Potato

The multielemental composition of organic and conventional winter wheat, spring barley, faba bean, and potato was analyzed with inductively coupled plasma-optical emission spectrometry (ICP-OES) and -mass spectrometry (ICP-MS). The crops were cultivated in two years at three geographically different field locations, each accommodating one conventional and two organic cropping systems. The conventional system produced the highest harvest yields for all crops except the nitrogen-fixing faba bean, whereas the dry matter content of each crop was similar across systems. No systematic differences between organic and conventional crops were found in the content of essential plant nutrients when statistically analyzed individually. However, chemometric analysis of multielemental fingerprints comprising up to 14 elements allowed discrimination. The discrimination power was further enhanced by analysis of up to 25 elements derived from semiquantitative ICP-MS. It is concluded that multielemental fingerprinting with semiquantitative ICP-MS and chemometrics has the potential to enable authentication of organic crops.

Effects of Organic and Conventional Growth Systems on the Content of Flavonoids in Onions and Phenolic Acids in Carrots and Potatoes

The demand for organic food products is steadily increasing partly due to the expected health benefits of organic food consumption. Polyphenols, such as flavonoids and phenolic acids, are a group of secondary plant metabolites with presumably beneficial health effects, and contents in plants are affected by, for example, plant nutrient availability, climate, pathogen infection, and pest attack. In the current study, onions, carrots, and potatoes were cultivated in two-year field trials in three different geographical locations, comprising one conventional and two organic agricultural systems. The contents of flavonoids and phenolic acids in plants were analyzed by pressurized liquid extraction and high-performance liquid chromatography-ultraviolet quantification. In onions and carrots, no statistically significant differences between growth systems were found for any of the analyzed polyphenols. On the basis of the present study carried out under well-controlled conditions, it cannot be concluded that organically grown onions, carrots, and potatoes generally have higher contents of health-promoting secondary metabolites in comparison with the conventionally cultivated ones.

Reproductive performance and bone status markers of gilts and lactating sows supplemented with two different forms of vitamin D.

In swine nutrition, little is known about the vitamin D requirements for reproductive processes and bone health. Consequently, the vitamin D recommendation for sows during gestation and lactation is not based on scientific reports. The current study was undertaken to obtain information on the dose-response pattern of 2 vitamin D sources, the commonly used cholecalciferol, called vitamin D(3), and a newly developed Hy.D product (25-hydroxycholecalciferol). In Exp. 1, a total of 160 gilts were randomly assigned from the first estrus until d 28 of gestation to dietary treatments containing 4 concentrations of 1 of the 2 different vitamin D sources [200, 800, 1,400, and 2,000 IU/kg of vitamin D from cholecalciferol or corresponding doses of 5, 20, 35, and 50 microg/kg of feed from 25(OH)D(3) (Hy.D)]. In a concurrent experiment, the same 8 dietary treatments were provided to 160 multiparous sows from the first day of mating until weaning. Plasma concentrations of 25(OH)D(3) were influenced by a dose x form interaction (P < 0.001); furthermore, plasma 25(OH)D(3) concentrations were influenced by the lactation state of the sows. Irrespective of the dietary dose and form of vitamin D provided to the sows, very little vitamin D was transferred to the progeny. Reproductive performance was not influenced by dietary vitamin D treatments, except for a decreased number of stillborn piglets (P = 0.03, SE = 0.40) with the larger doses of vitamin D (1,400 and 2,000 IU of vitamin D, resulting in 1.17 and 1.13 stillborn piglets per litter, respectively) compared with the smaller doses of vitamin D (200 and 800 IU of vitamin D, resulting in 1.98 and 1.99 stillborn piglets per litter, respectively). In the gilt trial, the ultimate strength of the bones (P = 0.01) and their content of ash (P = 0.02) were greater when vitamin D(3) was supplemented in doses larger than 800 IU, compared with the same amount of Hy.D supplementation. In the sow experiment, lactation day (P < 0.001), rather than dietary vitamin D, influenced the concentrations of osteocalcin and Ca as well as the activities of total alkaline phosphatase and bone alkaline phosphatase in plasma. Age of the suckling piglets affected their plasma bone health markers. In conclusion, at doses greater than 200 IU, Hy.D was more bioavailable than vitamin D(3) and, as such, could be considered an equivalent or even more advantageous source of vitamin D. In addition, a dietary dose of approximately 1,400 IU of vitamin D is recommended for reproducing swine. Irrespective of the dietary dose and form of vitamin D provided to the sows, very little vitamin D was transferred to the progeny.

Comparison of polyacetylene content in organically and conventionally grown carrots using a fast ultrasonic liquid extraction method.

A rapid and sensitive analytical method for quantification of polyacetylenes in carrot roots was developed. The traditional extraction method (stirring) was compared to a new ultrasonic liquid processor (ULP)-based methodology using high-performance liquid chromatography-ultraviolet (HPLC-UV) and mass spectrometry (MS) for identification and quantification of three polyacetylenes. ULP was superior because a significant reduction in extraction time and improved extraction efficiencies were obtained. After optimization, the ULP method showed good selectivity, precision [relative standard deviations (RSDs) of 2.3-3.6%], and recovery (93% of falcarindiol) of the polyacetylenes. The applicability of the method was documented by comparative analyses of carrots grown organically or conventionally in a 2 year field trial study. The average concentrations of falcarindiol, falcarindiol-3-acetate, and falcarinol in year 1 were 222, 30, and 94 mug of falcarindiol equiv/g of dry weight, respectively, and 3-15% lower in year 2. The concentrations were not significantly influenced by the growth system, but a significant year-year variation was observed for falcarindiol-3-acetate.

Is it really organic? - Multi-isotopic analysis as a tool to discriminate between organic and conventional plants

Novel procedures for analytical authentication of organic plant products are urgently needed. Here we present the first study encompassing stable isotopes of hydrogen, carbon, nitrogen, oxygen, magnesium and sulphur as well as compound-specific nitrogen and oxygen isotope analysis of nitrate for discrimination of organically and conventionally grown plants. The study was based on wheat, barley, faba bean and potato produced in rigorously controlled long-term field trials comprising 144 experimental plots. Nitrogen isotope analysis revealed the use of animal manure, but was unable to discriminate between plants that were fertilised with synthetic nitrogen fertilisers or green manures from atmospheric nitrogen fixing legumes. This limitation was bypassed using oxygen isotope analysis of nitrate in potato tubers, while hydrogen isotope analysis allowed complete discrimination of organic and conventional wheat and barley grains. It is concluded, that multi-isotopic analysis has the potential to disclose fraudulent substitutions of organic with conventionally cultivated plants.

A Randomized Controlled Trial Comparing Telemedical and Standard Outpatient Monitoring of Diabetic Foot Ulcers

OBJECTIVE The role of telemedical monitoring in diabetic foot ulcer care is still uncertain. Our aim was to compare telemedical and standard outpatient monitoring in the care of patients with diabetic foot ulcers in a randomized controlled trial. RESEARCH DESIGN AND METHODS Of the 736 screened individuals with diabetic foot ulcers, 401 met the eligibility criteria and were randomized between October 2010 and November 2014. The per-protocol telemedical monitoring consisted of two consultations in the patient’s own home and one consultation at the outpatient clinic. Standard practice consisted of three outpatient clinic visits. The three-visit cycle was repeated until study end point. The study end points were defined as complete ulcer healing, amputation, or death. RESULTS One hundred ninety-three individuals were randomized to telemedical monitoring and 181 to standard care. Demographics were similar in both groups. A cause-specific Cox proportional hazards model showed no difference in individuals monitored through telemedicine regarding wound healing (hazard ratio 1.11 [95% CI 0.87, 1.42], P = 0.42) or amputation (0.87 [0.54, 1.42], P = 0.59). We found a higher mortality incidence in the telemedical monitoring group compared with the standard outpatient monitoring group (8.68 [6.93, 10.88], P = 0.0001). CONCLUSIONS The findings of no significant difference regarding amputation and healing between telemedical and standard outpatient monitoring seem promising; however, for telemedical monitoring, a higher mortality throws into question the role of telemedicine in monitoring diabetic foot ulcers. Further studies are needed to investigate effects of telemedicine on mortality and other clinical outcomes and to identify patient subgroups that may have a poorer outcome through telemedical monitoring.

Heritability of metoprolol and torsemide pharmacokinetics

Genetic variation in the pharmacokinetics of metoprolol and torsemide due to polymorphisms in CYP2D6, CYP2C9, and OATP1B1 has been extensively studied. However, it is still unknown how much of the variation in pharmacokinetics of these two clinically important drugs in total is due to genetic factors. Metoprolol and torsemide were intravenously administered to 44 monozygotic and 14 dizygotic twin pairs. Metoprolol area under the curve (AUC) varied 4.7‐fold and torsemide AUC 3.5‐fold. A very high fraction of AUC variations, 91% of metoprolol and 86% of torsemide, were found to be due to additive genetic effects. However, known genetic variants of CYP2D6, ‐2C9, and OATP1B1 explained only 39%, 2%, and 39% of that variation, respectively. Comparable results for genetically explained variation in pharmacokinetics and pharmacodynamics have been found for other substrates of these enzymes earlier. These findings indicate that a substantial fraction of the heritable variability in the pharmacokinetics of metoprolol and torsemide remains to be elucidated.

The Heritability of Breast Cancer among Women in the Nordic Twin Study of Cancer

Background: Family history is an established risk factor for breast cancer. Although some important genetic factors have been identified, the extent to which familial risk can be attributed to genetic factors versus common environment remains unclear. Methods: We estimated the familial concordance and heritability of breast cancer among 21,054 monozygotic and 30,939 dizygotic female twin pairs from the Nordic Twin Study of Cancer, the largest twin study of cancer in the world. We accounted for left-censoring, right-censoring, as well as the competing risk of death. Results: From 1943 through 2010, 3,933 twins were diagnosed with breast cancer. The cumulative lifetime incidence of breast cancer taking competing risk of death into account was 8.1% for both zygosities, although the cumulative risk for twins whose co-twins had breast cancer was 28% among monozygotic and 20% among dizygotic twins. The heritability of liability to breast cancer was 31% [95% confidence interval (CI), 10%–51%] and the common environmental component was 16% (95% CI, 10%–32%). For premenopausal breast cancer these estimates were 27% and 12%, respectively, and for postmenopausal breast cancer 22% and 16%, respectively. The relative contributions of genetic and environmental factors were constant between ages 50 and 96. Our results are compatible with the Peto–Mack hypothesis. Conclusion: Our findings indicate that familial factors explain almost half of the variation in liability to develop breast cancer, and results were similar for pre- and postmenopausal breast cancer Impact: We estimate heritability of breast cancer, taking until now ignored sources of bias into account. Cancer Epidemiol Biomarkers Prev; 25(1); 145–50. ©2015 AACR.

Paternal age and telomere length in twins: the germ stem cell selection paradigm

Telomere length, a highly heritable trait, is longer in offspring of older fathers. This perplexing feature has been attributed to the longer telomeres in sperm of older men and it might be an ‘epigenetic’ mechanism through which paternal age plays a role in telomere length regulation in humans. Based on two independent (discovery and replication) twin studies, comprising 889 twin pairs, we show an increase in the resemblance of leukocyte telomere length between dizygotic twins of older fathers, which is not seen in monozygotic twins. This phenomenon might result from a paternal age‐dependent germ stem cell selection process, whereby the selected stem cells have longer telomeres, are more homogenous with respect to telomere length, and share resistance to aging.

Efficacy and safety of fosaprepitant for the prevention of nausea and emesis during 5 weeks of chemoradiotherapy for cervical cancer (the GAND-emesis study): a multinational, randomised, placebo-controlled, double-blind, phase 3 trial

BACKGROUND The role of the neurokinin-1 (NK-1) receptor antagonists in the prevention of radiation-induced nausea and vomiting has not been established. The purpose of the GAND-emesis study was to investigate the efficacy and safety of fosaprepitant in combination with palonosetron and dexamethasone in the prevention of nausea and vomiting during 5 weeks of fractionated radiotherapy and concomitant weekly cisplatin in patients with cervical cancer. METHODS This investigator initiated, multinational, randomised, double-blind, placebo-controlled phase 3 trial, included women with cervical cancer scheduled to receive fractionated radiotherapy and weekly cisplatin 40 mg/m(2) for 5 weeks. Patients had to be naive to chemotherapy and radiotherapy. Patients were randomly assigned to receive either single doses of fosaprepitant 150 mg intravenously or placebo (saline) in combination with palonosetron 0·25 mg intravenously and dexamethasone 16 mg orally before cisplatin administration. Randomisation was done by the unmasked pharmacist, who used a list of six numbers (a block) provided in a sealed envelope. A web-based randomisation number generator was used to generate the full list of randomisation numbers that was split up in blocks of six numbers. All patients received oral dexamethasone 8 mg twice a day on day 2, 4 mg twice a day on day 3, and 4 mg once on day 4. The treatment was repeated for 5 weeks. The primary endpoint was the proportion of patients with sustained no emesis after 5 weeks of treatment. The modified intention-to-treat population (all patients who received study medication) was used for the statistical analyses. The study was registered with ClinicalTrials.gov, number NCT01074697. FINDINGS Between June 15, 2010, and March 8, 2015, 246 patients from four countries consented to the study and were randomly assigned. Of these, 234 patients were eligible, having received study medication (118 received fosaprepitant, 116 received placebo). The proportion of patients with sustained no emesis at 5 weeks (competing risk analysis) was 48·7% (95% CI 25·2-72·2) for the placebo group compared with 65·7% (42·2-89·2) of patients for the fosaprepitant group. There was a significantly lower cumulative risk of emesis in the fosaprepitant group compared with the placebo group (subhazard ratio 0·58 [95% CI 0·39-0·87]; p=0·008). Treatments were generally well tolerated with few grade 3 adverse events none of which were related to the study treatment; the most common grade 3 adverse event during the 5 weeks of treatment was diarrhoea (11 [9%] of 118 patients in the fosaprepitant group vs six [5%] of 116 patients in the placebo group). There was only one report of a grade 4 adverse event (neutropenia), in the fosaprepitant group. No deaths were recorded in either group. INTERPRETATION To our knowledge, this is the first study to investigate safety and efficacy of a NK-1 receptor antagonist during 5 weeks of radiotherapy and concomitant weekly cisplatin. Patients receiving fosaprepitant in addition to palonosetron and dexamethasone were less likely to experience emesis and nausea compared with those receiving palonosetron and dexamethasone alone. Both treatments were safe and well tolerated. Further investigations in other radiotherapy settings are warranted. FUNDING Private and hospital or university funding, unrestricted grants from Biovitrum and Helsinn Healthcare SA.