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J Böhm

Publications and source records attributed to J Böhm.

At least 19 recordsLinked to original sources

Dietary inulin affects the morphology but not the sodium-dependent glucose and glutamine transport in the jejunum of broilers.

Inulin, a prebiotic, is a fermentable oligosaccharide that may affect the intestinal mucosal architecture and the electrophysiological parameters. The effects of a diet with added inulin were tested on the jejunal morphology and electrogenic transport of Glc and Gln from the jejunal mucosa in broilers. Short-circuit current and transmucosal tissue resistance of jejunal flaps were measured in Ussing chambers. The feeding experiment was carried out in broilers (n = 40) using 1% inulin with an application period of 5 wk. The inulin-containing diet resulted in longer jejunal villi (P < 0.05) and deeper crypts (P < 0.01) than in control birds without affecting villus:crypt depth. Basal short-circuit current value remained unaffected by dietary treatment. Inulin supplementation did not modify the electrogenic transport of Glc and Gln in the jejunal mucosa. The basal value of transmucosal tissue resistance was significantly lower (P < 0.001) in the inulin-fed group compared with the control group. In conclusion, inulin supplementation affected the jejunal mucosal architecture but did not modify the electrogenic transport of Glc and amino acid under present experimental condition.

Animal Feed↗

In vitro effects of deoxynivalenol on small intestinal D-glucose uptake and absorption of deoxynivalenol across the isolated jejunal epithelium of laying hens.

Deoxynivalenol (DON) is a common mycotoxin contaminant in feedstuffs. It has been shown to cause diverse toxic effects in animals. The aim of the present study was to evaluate the effects of DON on the glucose transport capacity in chickens' jejunum and to investigate the permeation of DON itself by the Ussing chamber technique. Glucose uptake into chicken jejunal epithelia was measured after the addition of 200 mumol/L of (14)C-labeled glucose to the mucosal solution. Glucose uptake under control condition was 3.28 +/- 0.53 nmol/cm(2) x min. The contribution of sodium glucose-linked transporter 1 (SGLT-1) to total glucose uptake was estimated by inhibiting SGLT-1 with phlorizin (100 micromol/L). In the presence of phlorizin, glucose uptake was reduced (P < 0.05) to 1.21 +/- 0.19 nmol/cm(2) x min. Deoxynivalenol decreased (P < 0.05) the glucose uptake in the absence of phlorizin to 1.81 +/- 0.24 nmol/cm(2) x min but had no additional effect on the glucose uptake in the presence of phlorizin (0.97 +/- 0.17 nmol/cm(2) x min). Mucosal-to-serosal permeation of DON was proportional to the initial DON concentration over a concentration range from 1 to 10 mug/mL on the mucosal side. Apparent permeability at 10 microg/mL of DON measured 60 to 90 min after DON application was 1.7 x 10(-05) cm/s. It can be concluded that DON (10 mg/L) decreases glucose uptake almost as efficiently as phlorizin. The similarity between the effects of phlorizin and DON on glucose uptake evidences their common ability to inhibit Na(+)-D-glucose cotransport. In addition to local effects, DON can be absorbed from the jejunum. A predominant part of DON passes across the chicken intestinal epithelium by passive diffusion, which is likely on the paracellular pathway. The results imply that the exposure to DON-contaminated feeds may negatively affect animal health and performance by local (i.e., inhibition of intestinal SGLT-1) and systemic effects.

Absorption↗

Biological and prognostic role of acid cysteine proteinase inhibitor (ACPI, cystatin A) in non-small-cell lung cancer.

BACKGROUND: Acid cysteine protease inhibitor (ACPI) is an intracellular protein, often linked to neoplastic changes in epithelium and thought to have an inhibitory role in malignant transformation. AIM: To analyse the expression and prognostic role of ACPI in non-small-cell lung cancer (NSCLC). METHOD: Histological samples from 199 patients with resected NSCLC were stained immunohistochemically for the expression of ACPI in normal and preneoplastic bronchial epithelium, and in various types of lung carcinomas. RESULTS: A normal bronchial epithelium showed positive staining for ACPI in the basal cells, whereas the upper two-thirds of the dysplastic epithelium was ACPI positive. High staining for ACPI was found in 74% (91/123) of squamous-cell carcinomas, whereas 16% (8/49) of adenocarcinomas and 30% of (8/27) large-cell carcinomas showed the high expression of ACPI (p<0.001). Among squamous-cell carcinomas, low expression of ACPI was correlated with poor tumour differentiation (p=0.032). In the whole tissue, reduced expression of ACPI was associated with tumour recurrence (p=0.024). In overall survival (OS) and disease-free survival (DFS) analyses, the histological type of the tumour (both p<0.001) and stage of the tumour (p=0.001, p=0.013, respectively) were related to patient outcome. Low expression of ACPI in tumour cells was associated with poor OS and DFS (p<0.041, p=0.004, respectively). In multivariate analysis, ACPI did not retain its prognostic value, whereas the traditional factors were the most important prognostic factors. CONCLUSIONS: ACPI expression is linked with the malignant transformation of the bronchial epithelium and predicts a risk of tumour recurrence as well as poor rate of survival for the patients. However, ACPI does not have any independent prognostic value in NSCLC.

Adenocarcinoma↗

[Extragenital endometriosis leading to piriformis syndrome].

We report on a 44-year-old woman with a history of sciatica fluctuating with her menstrual cycle and going back over 10 years; ultimately it was present continuously and became disabling. Over the years the patient developed ipsilateral foot-drop, a sensory disorder in the lateral aspect of the lower limb and back of the foot, and atrophy of the gluteus muscle. MRI confirmed the suspicion of extragenital endometriosis, which had caused piriformis syndrome by compression with consequent damage to the sciatic and inferior gluteal nerves. After hormonal therapy had been tried without success, the endometrioma was excised to relieve the pressure on the nerves, and the diagnosis was confirmed histopathologically. The motor deficit remained up to the 15 months since surgery, but the patient is now free of pain.

Adult↗

Effect of addition of a probiotic microorganism to broiler diets contaminated with deoxynivalenol on performance and histological alterations of intestinal villi of broiler chickens.

An experiment was conducted to study the effects of deoxynivalenol (DON) on the performance of broilers, organ weights, and intestinal histology and to evaluate the efficacy of a probiotic feed additive (PB, Eubacterium sp.) with the ability to deepoxidize DON. Two hundred seventy-seven 1-d-old broiler chicks were randomly assigned to 1 of the 3 dietary treatments for 6 wk. The dietary treatments were 1) control; 2) artificially contaminated diets with 10 mg of DON/kg of diet; 3) DON-contaminated diets plus probiotic feed additive (DON-PB). The BW and the efficiency of feed utilization were not adversely affected (P > 0.05) by the inclusion of DON in the diets. A slight improvement in feed intake and BW gain over the course of the experiment was observed in broilers fed DON-PB with no change in feed efficiency. The absolute or relative organ weights were not altered (P > 0.05) in broilers fed the diet containing DON compared with controls and the DON-PB group. The absolute liver weights were numerically increased (P < 0.1) for broilers receiving the diet containing DON-PB. There were no significant differences in the absolute and relative weights of the gizzard, duodenum, pancreas, heart, and spleen. However, the absolute and relative weights of the jejunum and cecum were increased for DON-PB-fed broilers compared with the controls and DON group. No pathological lesions were found in the gut of birds fed DON-contaminated diets during the feeding trial, but mild intestinal changes were observed. The DON altered small intestinal morphology, especially in the duodenum and jejunum, where villi were shorter and thinner (P < 0.05). The addition of the eubacteria to the DON-contaminated feed of the broilers effectively alleviated the histological alterations caused by DON and led to comparable villus length as in the control group. In conclusion, diets with DON contamination below levels that induce a negative impact on health and performance could affect small intestinal morphology in broilers. The histological alterations caused by DON were reduced by supplementing the DON-containing diets with PB. This indicates that in case of DON contamination of feedstuffs, the addition of PB would be a proper way to counteract the possible effects caused by this mycotoxin.

Animals↗

Effects of feeding deoxynivalenol contaminated wheat on growth performance, organ weights and histological parameters of the intestine of broiler chickens.

A feeding trial was conducted to evaluate the effects of moderate dietary concentrations of deoxynivalenol (DON) during a 21-day feeding experiment on the performance of broilers. Fifteen 1-day-old broiler chicks were randomly divided into two groups. The control group was fed non-contaminated diet. Another group of broilers was fed a diet naturally contaminated with 5 mg DON/kg diet. Deoxynivalenol had no effect (p > 0.05) on feed consumption, feed conversion, body-weight gain, live body weight or mortality. The absolute and relative weight of the organs (gizzard, pancreas, heart, spleen, colon and caecum) were not altered by the dietary inclusion of DON contaminated grain. However, both the absolute and relative weight of small intestine was decreased (p < 0.01) in DON fed broilers compared to the controls. No gross lesions were detected in any of the organs of birds fed contaminated wheat during the feeding trial. The microscopic examination revealed that, the height and the width of villi in duodenum decreased (p < 0.05) in birds fed DON contaminated wheat compared to controls. On the other hand the height and the width of jejunum villi were not affected (p > 0.05). This study indicates that feeding DON for 21 days to broiler chickens at a concentration of up to 5 mg/kg of diet influenced the weight of the small intestine as well as intestinal histology, especially the duodenum, as evidenced by shorter and thinner villi. In conclusion, diets with DON contamination below levels that induce negative impact on health and performance could affect small intestinal morphology in broilers.

Animal Feed↗

Functional expression of the renin-angiotensin system in human podocytes.

Experimental and clinical studies impressively demonstrate that angiotensin-converting enzyme inhibitors (ACEI) and angiotensin receptor blockers (ARB) significantly reduce proteinuria and retard progression of glomerular disease. The underlying intraglomerular mechanisms are not yet fully elucidated. As podocyte injury constitutes a critical step in the pathogenesis of glomerular proteinuria, beneficial effects of ACEI and ARB may partially result from interference with a local renin-angiotensin system (RAS) in podocytes. The knowledge of expression and function of a local RAS in podocytes is limited. In this study, we demonstrate functional expression of key components of the RAS in differentiated human podocytes: podocytes express mRNA for angiotensinogen, renin, ACE type 1, and the AT(1) and AT(2) angiotensin receptor subtypes. In Western blot experiments and immunostainings, expression of the AT(1) and AT(2) receptor was demonstrated both in differentiated human podocytes and in human kidney cortex. ANG II induced a concentration-dependent increase in cytosolic Ca(2+) concentration via AT(1) receptors in differentiated human podocytes, whereas it did not increase cAMP. Furthermore, ANG II secretion was detected, which was blocked by neither the ACEI captopril nor the renin inhibitor remikiren nor the chymase inhibitor chymostatin. ANG II secretion of podocytes was not increased by mechanical stress. Finally, ANG II was found to increase staurosporine-induced apoptosis in podocytes. We speculate that ACEI and ARB exert their beneficial effects, in part, by interfering with a local RAS in podocytes. Further experiments are required to identify the underlying molecular mechanism(s) of podocyte protection.

Angiotensinogen↗

In vitro effects of deoxynivalenol on electrical properties of intestinal mucosa of laying hens.

Deoxynivalenol (DON) is common in European cereal grains, and of all the trichothecenes, poses the greatest problems to animal health. The present study investigated the effects of DON on electrophysiological parameters in laying hens' jejunum mounted in Ussing chambers. In vitro studies were performed to measure the effects of different luminal concentrations of DON (0.5, 1, 5, and 10 microg/mL) on the transmural potential difference, electrical tissue resistance, and electrogenic ion flux rates (short-circuit current, Isc) across the isolated gut mucosa. Deoxynivalenol did not alter (P > 0.05) the transmural potential difference. Resistance was higher (P < 0.05) in the tissues exposed to DON compared with basal values. Deoxynivalenol caused a dose-dependent decrease in Isc (P < 0.05). To investigate the mechanism of action of DON, amiloride (a specific inhibitor for Na+ transport) was added after incubation of the tissue with DON. Amiloride did not decrease (P > 0.05) Isc under these conditions. This may indicate that DON inhibited the Na+ transport before addition of amiloride, which did not then show further inhibitory effects. The addition of D-glucose (5 mmol/L) on the luminal side of the isolated mucosa increased (P < 0.05) Isc, and this effect was reversed by phlorizin (a specific inhibitor of sodium/glucose transporter 1), indicating that the glucose-induced Isc increase may be due to Na+-D-glucose cotransport. In our study, DON decreased (P < 0.05) the glucose-induced Isc in a similar way to phlorizin. The remarkable similarity between the effects of phlorizin and DON on electrical properties seemed to be consistent with their common ability to inhibit Na+-D-glucose cotransport. In conclusion, DON decreased the Isc via inhibition of Na+ transport. The effect on intestinal electrical properties was similar to that of phlorizin after addition of glucose, suggesting that DON may inhibit Na+-D-glucose cotransport. The inhibition of Na+ transport and Na+-D-glucose cotransport are important mechanisms of DON toxicity in the intestine of laying hens.

Amiloride↗

Effects of luminal deoxynivalenol and L-proline on electrophysiological parameters in the jejunums of laying hens.

Most amino acids are cotransported with sodium. Deoxynivalenol (DON) decreases glucose absorption in the chicken small intestine in vivo and in vitro, and this effect is apparently mediated by the inhibition of the sodium D-glucose cotransporter. DON could selectively modulate the activities of other intestinal transporters. In order to assess this hypothesis, a study was conducted to characterize the in vitro effects of DON in the presence of mucosal amino acids, using L-proline as a model, on the electrophysiological parameters in the jejunums of laying hens. L-Proline (mucosal concentration of 1 mmol/L) was added to a stripped proximal part of jejunum sheets mounted in Ussing chambers in Ringer buffer, and the electrical properties were measured. The transmural potential difference (PD) was nearly constant between the treatments. The tissue resistance (Rt) was higher (P < 0.05) in the tissues exposed to DON compared with basal values and the values after addition of L-proline. Addition of L-proline on the luminal side of the isolated mucosa increased (P < 0.05) the short circuit-current (Isc), and it decreased (P < 0.05) after addition of DON, indicating that the proline-induced Isc was altered by DON. The addition of proline after incubation of the tissues with DON had no effect (P > 0.05) on PD or Rt. Proline did not increase the Isc under these conditions. DON decreased (P < 0.1) the Isc after addition of proline, indicating that DON inhibited the Na+-amino acid co-transport. We concluded from the present study that the amino acid cotransporter activity appears to be highly sensitive to DON suppression.

Animals↗

Elevated expression of hyaluronan and its CD44 receptor in the duodenal mucosa of coeliac patients.

AIMS: Since hyaluronan (HA) metabolism is disturbed in some malignant tumours and in inflammatory diseases, we analysed HA and its receptor CD44 as well as the expression of the Ki67 nuclear protein, a marker of cell proliferation, in histological sections of duodenal biopsies of coeliac disease patients and controls. METHODS AND RESULTS: The study group consisted of 52 patients with coeliac disease in remission, 40 patients with newly diagnosed disease and 10 healthy control subjects. HA was detected with a specific biotinylated probe prepared from cartilage aggrecan and link protein, and CD44 with an antibody recognizing all forms of CD44 and another specific for its v6 variant. For the expression of the nuclear protein, monoclonal antibody MIB-1 was used. The percentage of HA-positive cells in surface epithelium was higher in newly diagnosed patients (13%) compared with patients in remission (11%) and controls (2%). In addition, HA intensity in the lamina propria was decreased in the newly diagnosed patients. In patients with active disease, 22-26% of the surface epithelium was CD44+, whereas the corresponding figure in patients in remission was 5%, and that of controls 1%. The more intensive MIB-1 labelling in the duodenal epithelium of coeliac patients without treatment was normalized after gluten-free diet. CONCLUSIONS: The HA-positive coat on surface epithelium seen even in patients in remission suggests persistent or even permanent changes in the epithelial permeability barrier in coeliac disease.

Adult↗

Effects of Fusarium toxin-contaminated wheat grain on nutrient turnover, microbial protein synthesis and metabolism of deoxynivalenol and zearalenone in the rumen of dairy cows.

The aim of the present study was to examine the effects of feeding Fusarium toxin-contaminated wheat to dairy cows on nutrient utilization in the rumen and on duodenal flow of deoxynivalenol (DON), zearalenone (ZON) and their metabolites. Six dairy cows fitted with a large rumen cannula and a simple T-shaped cannula at the proximal duodenum was used in two experiments. The experiments included a control period in which the uncontaminated control wheat was fed and a period in which the control wheat was replaced by the Fusarium toxin-contaminated wheat (8.05 and 7.15 mg DON/kg and 0.26 and 0.1 mg ZON/kg in Expts 1 and 2 respectively). The wheat portion of the daily ration amounted to 50% on a dry matter (DM) basis and rations were completed with hay or grass silage. Five of the six cows were non-lactating and the total daily DM-intake ranged between 4 and 12 kg. The pH-values and the concentration of volatile fatty acids in ruminal fluid were not significantly influenced by feeding the contaminated wheat. In contrast, the postprandial ammonia concentration was consistently higher when the mycotoxin-contaminated wheat was fed. Moreover, the flow of microbial protein and utilizable protein at the duodenum were reduced at the same time. The concentrations of DON and ZON and of their metabolites in freeze-dried duodenal digesta were either not detectable or negligible during the control periods whereas distinct concentrations were measured during the periods where the contaminated wheat was fed. DON was nearly completely metabolized to de-epoxy-DON and the flow at the duodenum ranged between 4% and 28% of DON-intake. The ZON metabolites alpha-zearalenol (ZOL) and beta-ZOL were recovered at the duodenum beside the parent toxin ZON. Their recovery as a percentage of ZON-intake ranged between 43% and 132%. In conclusion, feeding of Fusarium toxin-contaminated wheat altered the ruminal protein utilization. The question of whether this effect was a result of the mycotoxin being present in the rumen or of Fusarium growth-related structural (cell wall) changes of the wheat grain needs to be clarified. The low recovery of DON at the duodenum would indicate either a nearly complete degradation of the molecule in the rumen or an absorption by the mucosa of the rumen, whereas the higher ZON recovery would suggest a lower degradation of the parent toxin in the rumen and/or recovery of some bile-originating entero-hepatic cycling ZON/metabolites.

Animal Feed↗

Cardiac magnetic resonance imaging: long term reproducibility of the late enhancement signal in patients with chronic coronary artery disease.

OBJECTIVE: To determine long term reproducibility of the late enhancement (LE) signal in contrast enhanced magnetic resonance imaging (MRI) and potential changes of the signal after revascularisation. METHODS: 33 patients (29 men, mean (SD) 61 (11) years) with coronary artery disease (CAD) and left ventricular dysfunction (ejection fraction 30 (7)%) underwent two contrast enhanced MRI procedures within 9 (3) months. Fifteen patients (group A: 14 men, 59 (12) years) had no interventions between the two studies. Eighteen patients underwent revascularisation after MRI 1 (group B: 15 men, 62 (9) years). Changes in the LE signal between the first and second MRIs were investigated in both groups as well as intraobserver and interobserver variabilities for delineation of the signal. RESULTS: The LE signal was highly reproducible in groups A and B for segmental analysis (concordance 86% v 82%, respectively; kappa = 0.70 v 0.67) and summed scores (group A: r = 0.97, p < 0.001; group B: r = 0.93, p < 0.001). The LE signal was quantified as 27 (27) cm3 in group A versus 30 (16) cm3 in group B in the first MRI and 26 (25) cm3 versus 30 (15) cm3, respectively, for the second MRI (both not significant). Moreover, low intraobserver and interobserver variabilities were observed in segmental analysis (kappa = 0.86 and 0.74, respectively, for group A, and kappa = 0.87 and 0.82, respectively, for group B). CONCLUSION: In patients with chronic CAD, the LE signal in contrast enhanced MRI is very stable over an extended time period. These results further characterise contrast enhanced MRI as a useful tool for myocardial viability assessment. Low intraobserver and interobserver variabilities promise robustness of the method for clinical application.

Aged↗

Effects of deoxynivalenol on general performance and electrophysiological properties of intestinal mucosa of broiler chickens.

A feeding trial was conducted to evaluate the effects of diets contaminated with deoxynivalenol (DON on the performance of broilers and on the electro-physiological parameters of the gut. The control group was fed the starter and finisher diets without addition of DON. Another group of broilers was fed the starter and finisher diets with 10 mg/kg DON, whereas another group was fed the DON-contaminated diets supplemented with a microbial feed additive (Eubacterium sp.). The diets were provided ad libitum for 6 wk. DON had no effect (P > 0.05) on feed consumption, feed conversion, or body weight. The effect of DON on the electrophysiological parameters of the jejunum was studied in vitro using isolated gut mucosa in Ussing chambers. At the end of the feeding period, 7 birds from each group were killed, and the basal and glucose stimulated transmural potential difference (PD), short-circuit current (Isc), and electrical resistance (R) were measured in the isolated gut mucosa to characterize the electrical properties of the gut. The transmural PD did not differ (P > 0.05) among groups. The tissue resistance was greater (P < 0.05) in birds receiving DON and the microbial feed additive than in the controls and DON group. Addition of D-glucose on the luminal side of the isolated mucosa increased (P < 0.05) Isc in the control and DON-probiotic (Eubacterium sp.; PB) groups, whereas it decreased (P < 0.05) in the DON group indicating that the glucose-induced Isc was altered by DON. Addition of the eubacteria to the DON contaminated feed of the broilers led to electrophysiological properties in the gut that were comparable with those of the control group. It could be concluded that 10 mg/kg DON in the diet impaired the Na(+)-D-glucose cotransport in the jejunum of broilers. In the absence of clinical signs, and without impaired performance, DON appeared to alter the gut function of broilers. The addition of Eubacterium sp. may be useful in counteracting the toxic effects of DON on intestinal glucose transport.

Animal Feed↗

Feeding laying hens seal blubber oil: effects on egg yolk incorporation, stereospecific distribution of omega-3 fatty acids, and sensory aspects.

Seventy-two 26-wk-old Single Comb White Leghorn laying hens were randomly assigned to 36 cages (2 per cage) in a 3-orthogonal 4 x 4 latin square, with the fourth row suppressed, to assess the effect of feeding refined seal blubber oil (SBO, containing 22.2% omega-3 fatty acids) on the fatty acid composition and position in the egg yolk lipids. The experiment was conducted over a period of 9 wk. Eggs were collected and numbered, and the weights were recorded for each week and cage. Eggs collected at wk 5 and 9 were used for total lipid, lipid class, fatty acid, and positional analyses. Sensory evaluation was carried out on eggs collected at wk 6 and 7. Feeding SBO at 1.25% led to an increase (P < 0.0001) in the long-chain omega-3 polyunsaturated fatty acids (LCn3PUFA) and a concomitant decrease (P < 0.0001) in arachidonic acid (ARA) in the egg yolk lipids. Yet this amount of SBO in the diet had no effect (P > 0.1) on the sensory attributes of the egg and on production parameters such as egg weight, number of eggs laid, and feed intake (P > 0.05). When feeding SBO in amounts higher than 1.25% proportionately, a plateau effect of the LCn3PUFA content of the eggs was observed. This appears to be because the PUFA content in the sn-2 position of the phospholipids cannot exceed a certain amount. When this amount is reached, the LCn3PUFA will be increasingly stored in triglycerides. The results presented here clearly indicate how eggs can be produced with optimized composition of LCn3PUFA without affecting (P > 0.1) the sensory properties of the eggs. The procedures elaborated herein provide directly applicable consequences for the food industry.

Animals↗

Adaptation of the present concept of dosimetric radiation protection quantities for external radiation to radiation protection practice.

The present concept of dosimetric radiation protection quantities for external radiation is reviewed. For everyday application of the concept some adaptations are recommended. The check of the compliance with dose limits should be performed either by the comparison with values of the respective operational quantities directly or by the calculation of the protection quantity by means of the operational quantity, the appertaining conversion coefficient and additional information of the radiation field. Only four operational quantities are regarded to be sufficient for most applications in radiation protection practice. The term equivalent should be used in the connection dose equivalent only. Proposals are made for names of frequently used operational quantities which are denoted up to now by symbols only.

Calibration↗

Simultaneous determination of major B-trichothecenes and the de-epoxy-metabolite of deoxynivalenol in pig urine and maize using high-performance liquid chromatography-mass spectrometry.

A selective analytical method based on high-performance liquid chromatography (HPLC), combined with atmospheric pressure chemical ionisation (APCI-) mass spectrometry (MS), has been developed for simultaneous determination of B-trichothecenes and the major metabolites of deoxynivalenol. The method allows simultaneous analysis of nivalenol (NIV), deoxynivalenol (DON), 15-acetyldeoxynivalenol (15-AcDON), 3-acetyldeoxynivalenol (3-AcDON), fusarenon X (Fus-X) and de-epoxydeoxynivalenol (DOM-1). The method is based on one-step sample clean-up using a multifunctional MycoSep column. A linear gradient mobile phase system, consisting of water:acetonitrile:methanol (H2O:ACN:MeOH) at a flow-rate of 1 ml/min, and a Polar-RP C18 column, were utilised to obtain the best resolution of all tested compounds along with column and equilibrating within 30 min. Dexamethasone (Dex) was used as internal standard. The developed method shows good repeatability for inter- and intra-day precisions as well as good linearity of calibration curves (r2 ranged from 0.9936 to 0.9998). Average recoveries for tested compounds in both matrices have been determined ranging from 63.7 to 102.3% and limit of quantification (LOQ) ranged from 25 to 150 ng/g. The utility and practical impact of the method is demonstrated using contaminated pig urine and maize samples.

Animals↗

Dioxin residues in the edible tissue of broiler chicken.

The aim of this study was to determine the contamination of broiler chicken with polychlorinated dibenzodioxins and dibenzofurans (PCDD/Fs) after feeding either uncontaminated feed or feed contaminated with 1, 2 or 4 ng/kg toxic equivalents (TEQ). The feed was mixed with pure substances of PCDD/Fs to get the intended contamination. Ten groups of seven 1-day-old chickens each were housed in special cages for broiler fattening. The fattening period lasted for 6 weeks. The contaminated feed was given for either 2, 4, or 6 weeks, one group received uncontaminated feed (control group). After slaughtering the edible parts of the chickens breast and leg including the skin were homogenized. Body weight gain and feed conversion (kg feed/kg body weight gain) were in the normal range (final weight 1.98+/-0.07 kg; feed conversion 1.74+/-0.03). One kilogram edible tissue contained an average of 21.2+/-4.1% of the total TEQ-intake in all groups. The PCDD/Fs residues in the edible tissues significantly correlated with the amount of PCDD/Fs-intake (r=0.99; Pearson correlation). There was no decrease in dioxin residues (% of total PCDD/Fs intake) after a 2 or 4 weeks withdrawal period. The results of this trial predict that a threshold value of 2 pg/g chicken fat can be met if the PCDD/Fs content in chicken feed is not higher than 0.4 ng/kg.

Animal Feed↗

Evidence of clonality in chronic neutrophilic leukaemia.

BACKGROUND: Chronic neutrophilic leukaemia (CNL) is a rare myeloproliferative disorder of elderly patients characterised by sustained neutrophilia and splenomegaly. The diagnosis of CNL requires the exclusion of BCR/ABL positive chronic myelogenous leukaemia (CML) and of leukaemoid reactions (LRs). The differentiation between CNL and LR is problematic because both conditions share similar morphological features; it is also important because patients with CNL generally have a poor prognosis. AIMS: To determine whether CNL and LR could be distinguished on the basis of different clonality patterns. METHODS: Blood samples from 52 women were studied using the human androgen receptor gene assay (HUMARA). RESULTS: Monoclonality was found in the neutrophils in all 17 patients with different myeloproliferative syndromes (MPSs), including those with CNL. In four of the patients with CNL, autologous T cells were also monoclonal, suggesting that they belonged to the neoplastic clone. This finding was in contrast to other MPSs in which T cells were almost always polyclonal. Of nine patients with clinically suspected LR, the neutrophils of five were polyclonal, whereas three patients had monoclonal neutrophils, suggesting that they might be in the process of developing an MPS. Among 26 healthy blood donors, 20 had polyclonal neutrophils and five showed skewed clonality patterns. One case of LR and one normal blood donor were scored "not informative" at the HUMARA locus. CONCLUSIONS: Clonality studies of blood neutrophils using HUMARA aid in distinguishing female patients with monoclonal CNL from those with LR. For the diagnosis of CNL, monoclonality of the neutrophils should be demonstrated whenever possible.

Adult↗