Field and laboratory findings following the large-scale use of intermediate type infectious bursal disease vaccines in Denmark.

Following a period of clinical outbreaks of very virulent infectious bursal disease virus (vvIBDV) in Denmark, the histological bursal lesion score (HBLS) was used on a national scale to screen broiler flocks vaccinated with intermediate IBD vaccines for lesions indicative of IBDV challenge. High lesion scores were detected in a high percentage of healthy and well performing flocks despite the lack of other indications of the presence of vvIBDV. RT-PCR and subsequent sequencing showed the frequent presence of H253Q and H253N IBDV strains that were genetically close to the sequence of the intermediate vaccines with a relative risk ratio of 13.0 (P < 0.0001) in intermediate vaccine A or B vaccinated flocks compared to unvaccinated flocks. The relevance of these H253Q and H253N strains was tested under experimental conditions using a protocol derived from the European Pharmacopoeia for safety of live IBD vaccines. The results confirmed the higher pathogenicity for the bursa of these strains compared to intermediate vaccines as well as the negative effect on antibody response to a Newcastle disease (ND) vaccination performed at the peak of the bursa damage. The efficacy of the ND vaccination was still 100% showing that the H253N and H253Q IBDV strains would be considered as safe vaccine viruses. In conclusion, the use of the HBLS to screen commercial broiler flocks vaccinated with intermediate IBD vaccines for the presence of vvIBDV does not seem to be a reliable method due to the frequent occurrence of H253N and H253Q strains in those flocks. For screening of IBD vaccinated flocks for the presence of vvIBDV or other field strains, the RT-PCR with subsequent sequencing seems to be most suitable.

Correlates of objectively measured physical activity in 5-6-year-old preschool children.

AIM: The aim of this study was to identify gender specific physical activity correlates in Danish preschool children, using a socioecological approach. METHODS: This was a cross-sectional study conducted in Odense, Denmark. The gender specific models were based on data from 174 boys and 177 girls, 5-6 years of age and enrolled in 40 randomly selected preschools. Percentage of total daily time spent in moderate and vigorously physical activity (MVPA) was measured using ActiGraph accelerometers over 5 preschool days and 2 days off. Thirty-nine potential correlates of child MVPA across 5 domains were tested for associations with gender specific MVPA. RESULTS: The gender specific models consistently identified motor coordination and the parents' perceptions of their child's activity preferences during free play to be positively associated with MVPA. Days with rain or no preschool attendance were negatively associated with MVPA. For boys, rural area and the size of the preschool playground were positively associated with MVPA, whereas for girls, it was the age and the relative preschool indoor area. CONCLUSION: Individual and overall environmental correlates of MVPA were identified. However, most correlates were not or only weakly related to MVPA. It is possible that the interaction with peers and surrounding adults might contribute the most to preschool child MVPA.

The occurrence of volatile and non-volatile N-nitrosamines in cured meat products from the Danish market.

Several epidemiological studies emphasize the consumption of processed meat products as a risk factor of colorectal cancer, linking N-nitrosamines (NAs) formed during nitrite curing to this cancer risk. The occurrence of volatile N-nitrosamines (VNAs) has over the years been intensively studied while the knowledge on the occurrence and toxicity of non-volatile N-nitrosamines (NVNAs) is still limited. Therefore, this study focuses on quantification of both VNAs and NVNAs in a large selection of processed meat products. For this purpose, a robust, specific and sensitive method allowing analysis of seven VNAs and two NVNAs was optimized and validated using kassler, sausage, and salami. The limit of quantification achieved was 0.1-0.5 ng·g-1 for most of the VNA, and 2.3-4.2 ng·g-1 for the NVNA. In one hundred commercial samples N-nitroso-thiazolidine-4-carboxylic acid (NTCA) was the most frequently detected (97 samples) among all target NAs and it was found at concentrations ranging from 3.1 ng·g-1 to 1660 ng·g-1. The samples contained relatively low mean levels of the individual VNAs (≤1 ng·g-1). The levels of N-nitrosodimethylamine (NDMA), N-nitrosopyrrolidine (NPYR), and N-nitrosopiperidine (NPIP) ranged from non-detectable to 3.8, 10.8 and 2.9 ng·g-1, respectively. A correlation between the detected residual levels of nitrite and/or nitrate and concentrations of individual NAs could not be demonstrated. Based on principle component analysis (PCA) some correlations between salami, sausage and bacon and NAs could be shown.

Axonal lesion-induced microglial proliferation and microglial cluster formation in the mouse.

Microglia are innate immune cells and form the first line of defense of the CNS. Proliferation is a key event in the activation of microglia in acute pathology, and has been extensively characterized in rats, but not in mice. In this study we investigated axonal-lesion-induced microglial proliferation and surface antigen expression in C57BL/6 mice. Transection of the entorhino-dentate perforant path projection results in an anterograde axonal and a dense terminal degeneration that induces a region-specific activation of microglia in the dentate gyrus. Time-course analysis showed activation of microglial cells within the first week post-lesion and cell counting demonstrated a significant 1.6-fold increase in microglial numbers 24 h post-lesion reaching a maximal 3.8-fold increase 3 days post-lesion compared with controls. Double staining for the microglial macrophage antigen-1 and the proliferation marker bromodeoxyuridine, injected 1 h prior to perfusion, showed that lesion-reactive microglia accounted for the vast majority of proliferating cells. Microglia proliferated as soon as 24 h after lesion and 25% of all microglial cells were proliferating 3 days post-lesion. Immunofluorescence double staining showed that most activated, proliferating microglia occurred in multicellular clusters and co-expressed the intercellular adhesion molecule-1 and the hematopoietic stem cell marker cluster of differentiation 34. In conclusion, this study extends observations of axonal lesion-induced microglial proliferation in rats to mice, and provides new information on early microglial proliferation and microglial cluster formation and surface antigen expression in the mouse.

Formation and mitigation of N-nitrosamines in nitrite preserved cooked sausages.

Literature on formation and mitigation of N-nitrosamine (NA) and especially non-volatile NA (NVNA) in meat products is scarce and the present study is therefore a relevant contribution to the field. We found positive correlation between the levels of N-nitrosopiperidine (NPIP), N-nitrosohydroxyproline (NHPRO), N-nitrosoproline (NPRO), N-nitrosothiazolidine-4-carboxylic acid (NTCA) and N-nitroso-2-methyl-thiazolidine-4-carboxylic acid (NMTCA) and the amount of nitrite added to cooked pork sausages. The levels studied were 0, 60, 100, 150, 250 and 350 mg kg(-)(1). The levels of N-nitrosodimethylamine (NDMA) and N-nitrosopyrrolidine (NPYR) remained at or below limit of quantification. Erythorbic acid inhibited the formation of NHPRO, NPRO, NPIP and NTCA. This inhibition was for NTCA and NMTCA counteracted by addition of free iron. Ascorbyl palmitate had less inhibitory effect than erythorbic acid and a combination of the two provided no further protection. Increasing the black pepper content increased the levels of NPIP and NMTCA. Only slight effects of increased fat content and addition of tripolyphosphate were observed.

Dietary exposure to volatile and non-volatile N-nitrosamines from processed meat products in Denmark.

Recent epidemiological studies show a positive association between cancer incidence and high intake of processed meat. N-nitrosamines (NAs) in these products have been suggested as one potential causative factor. Most volatile NAs (VNAs) are classified as probable human carcinogens, whereas the carcinogenicity for the majority of the non-volatile NA (NVNA) remains to be elucidated. Danish adults (15-75 years) and children (4-6 years) consume 20 g and 16 g of processed meat per day (95th percentile), respectively. The consumption is primarily accounted for by sausages, salami, pork flank (spiced and boiled) and ham. This consumption results in an exposure to NVNA of 33 and 90 ng kg bw(-1) day(-1) for adults and children, respectively. The exposure to VNA is significantly lower amounting to 0.34 and 1.1 ng kg bw(-1) day(-1) for adults and children, respectively. Based on a BMDL10 of 29 µg kg bw(-1) day(-1) a MOE value ≥17,000 was derived for the exposure to NA known to be carcinogenic (VNA including NSAR), indicating an exposure of low concern. The exposure to the NVNA is substantially higher and if found to be of toxicological significance the exposure may be of concern.

Prediction of relapse with the TRH test and prophylactic amitriptyline in 39 patients with endogenous depression.

The authors conducted a double-blind prospective study of 39 patients with unipolar endogenous depression who recovered after ECT. Thyrotropin (TSH)-releasing hormone (TRH) tests were performed before and after ECT. Patients were divided into three groups on the basis of their altered TSH response: Persistent remission was predicted for patients in group 1 (N = 15) and relapse was predicted for groups 2A (N = 13) and 2B (N = 11). Patients in groups 1 and 2A received placebo and those in group 2B received amitriptyline for 6 months. Fewer relapses occurred in groups 1 and 2B than in group 2A (p less than .05), showing that relapse can be predicted by the TRH test and prevented by amitriptyline.

Phosphorylation of proteins in chick ciliary ganglion under conditions that induce long-lasting changes in synaptic transmission: phosphoprotein targets for nitric oxide action.

Production of nitric oxide and the activation of protein kinases are required for long-term potentiation of synaptic transmission at the giant synapses in chicken ciliary ganglion. In the present study, we investigated the ability of nitric oxide to regulate the phosphorylation of endogenous proteins under conditions that induced long-term potentiation in intact ciliary ganglion and the protein kinases responsible for the phosphorylation of these proteins in lysed ciliary ganglion. Using Calcium Green-1 we showed that the nitric oxide donor sodium nitroprusside did not change the intraterminal Ca2+ dynamics in ciliary ganglion. Two dimensional phosphopeptide analysis of 32Pi-labelled intact ciliary ganglion showed that the sodium nitroprusside (300 microM) increased the phosphorylation of several phosphopeptides (P50a, P50b and P41) derived from proteins at 50,000 and 41,000 mol. wts which we have called nitric oxide-responsive phosphoproteins. A similar stimulation of phosphorylation was achieved by 8-bromo-cyclic AMP (100 microM), which also induced long-term potentiation, but not by phorbol dibutyrate (2 microM) that does not induce long-term potentiation in ciliary ganglion. When subcellular fractions from lysed ciliary ganglion were labelled in vitro by [gamma-32P]ATP in the presence of purified cGMP-dependent, cAMP-dependent or Ca2+-phospholipid-dependent protein kinases, we identified cyclic GMP-dependent protein kinase substrates that gave rise to phosphopeptides co-migrating with P50a, P50b and P41 from 32Pi-labelled intact ciliary ganglion. P50a and P41 were derived from soluble proteins while P50b was derived from a membrane-associated protein. The proteins giving rise to P50a, P50b and P41 were also substrates for cyclic AMP-dependent protein kinase, but not for calcium and phospholipid-dependent protein kinase in vitro, suggesting that nitric oxide-responsive phosphoproteins are convergence points in information processing in vivo and their phosphorylation might represent an important mechanism in nitric oxide-mediated synaptic plasticity in ciliary ganglion.

Automatic prediction of platelet utilization by time series analysis in a large tertiary care hospital.

The authors investigated the potential of various time series models to aid in the prediction of platelet utilization patterns at the authors' institution. During a seven-week feasibility study, prediction accuracy of time series models was slightly inferior to that of an experienced platelet transfusion coordinator. The models that adjusted for day-of-the-week variability were superior to simpler models in a prospective evaluation over a three-month period. Conversion to a time series model for platelet utilization prediction has resulted in significant savings in personnel costs for management of platelet concentrate inventory in the blood bank. Outdate rates have been lower than those achieved by the platelet transfusion coordinator, with no sacrifice in availability occurring during the evaluation period.

Prevention and management of drug-induced peripheral neuropathy.

When symptoms of peripheral neuropathy appear, the possibility that they have been induced by drugs should be considered. A large number of drugs of all kinds, several of which are considered indispensable, have been implicated in peripheral neuropathy. A list of some of these drugs is provided. Neuropathy is a universal and dose-limiting factor during treatment with vinca alkaloids, but is otherwise a rare complication of drug therapy. Drug-induced peripheral neuropathy is almost always due to a dose-dependent primary axonal degeneration caused either by toxic reactions or by metabolic changes in neurons or their surroundings. The use of drugs should be restricted, especially in patients with a risk for development of neuropathy or with already existing neuropathy, e.g. patients with hepatic or renal failure, diabetes mellitus, or malnutrition. Patients should be given vitamins, prophylactically or therapeutically, which will sometimes allow a treatment to be continued. In other cases of drug-induced neuropathy the drug should be stopped. Reversal depends on the severity of the neuropathy, intensity and duration of the treatment and existence of causative cofactors, but generally the prognosis is good. While waiting for recovery physiotherapy is of importance, and when paraesthesia and pain are troublesome the patient should be treated with carbamazepine, imipramine or lidocaine (lignocaine).

Effect of amitriptyline on the thyrotropin response to thyrotropin-releasing hormone in endogenous depression.

Patients with endogenous depression whose depressive episodes were clinically resolved after electroconvulsive therapy were divided into two groups: one in which patients remained well (n = 16) and another in which patients relapsed within 6 months (n = 11). Treatment with amitriptyline for 3 weeks did not affect the median thyrotropin (thyroid-stimulating hormone; TSH) response to thyrotropin-releasing hormone (TRH) in recovered patients, whereas that in relapsed patients was significantly enhanced. The data suggest that amitriptyline affects the TSH response to TRH differently in stably recovered and relapsed patients. If this effect is maintained beyond the 3-week period studied, treatment with amitriptyline will invalidate the predictive value of the TRH test.

Purification and characterisation of a malto-oligosaccharide-forming amylase active at high pH from Bacillus clausii BT-21.

Bacillus clausii BT-21 produced an extracellular malto-oligosaccharide-forming amylase active at high pH when grown on starch substrates. The enzyme was purified to homogeneity by affinity and anion-exchange chromatography. The molecular weight of the enzyme estimated by sodium dodecyl sulfate polyacrylamide electrophoresis was 101 kDa. The enzyme showed an optimum of activity at pH 9.5 and 55 degrees C. Maltohexaose was detected as the main initially formed starch hydrolysis product. Maltotetraose and maltose were the main products obtained after hydrolysis of starch by the enzyme for an extended period of time and were not further degraded. The enzyme readily hydrolysed soluble starch, amylopectin and amylose, while cyclodextrins, pullulan or dextran were not degraded. The mode of action during hydrolysis of starch indicated an exo-acting type of amylolytic enzyme mainly producing maltohexaose and maltotetraose. Amino acid sequencing of the enzyme revealed high homology with the maltohexaose-forming amylase from Bacillus sp. H-167.

Suitability and limitations of methods for characterisation of activity of malto-oligosaccharide-forming amylases.

The suitability and limitations of essential methods and reference substrates used for characterisation of activity of amylolytic enzymes is investigated. Saccharogenic, chromogenic and chromatographic methods are included. The results are discussed in relation to the measurement of reaction rates, determination of action mode and product specificity and the impact on identification and nomenclature of malto-oligosaccharide-forming amylases. An accurate determination of reaction rates using the saccharogenic methods strongly depends on the degree of polymerisation (DP) of the standards used and the hydrolysis products formed by the amylase. Particularly the use of glucose as standard can lead to overestimates due to the differences in the reducing potential of glucose and malto-oligosaccharides. The reliability of the chromogenic methods for determination of action mode depends on the DP of the substrate and the specificity of the amylase. For a characterisation of the starch hydrolysis products and the variation in the DP during hydrolysis, high performance anion-exchange chromatography with pulsed amperometric detection provided a fast and reliable method. A literature survey revealed varying and inconsistent use of nomenclature of malto-oligosaccharide forming amylases. Therefore a systematic approach identifying three main classes of activity is suggested using not only the mode of action and the DP of the major product but also the stage of hydrolysis at which this product is formed.

The etiology and possible treatment of chronic fatigue syndrome/fibromyalgia.

It is suggested that chronic fatigue syndrome/fibromyalgia is caused by virus injury to the calcium channels leading to larger quantities than usual of calcium ions entering the striated muscle cells. Should this be true, then treatment with a calcium antagonist (CA) may possibly be of value.

[Poisoning with the brown roll-rim mushroom, Paxillus involutus]. / Forgiftning med svampen almindelig netbladhat, Paxillus involutus.

Brown Roll-Rim (Paxillus involutus) is a dangerous poisonous mushroom with symptoms of poisoning which occur a few hours after consumption. Repeated consumption may cause sensitization to a heat-stable toxin, resulting in haemolysis which may be of all degrees. In addition, severe gastrointestinal symptoms may be due to a heat-instable toxin. In cases with immune haemolysis, treatment consists of corticosteroid and possibly plasmapheresis and symptomatic therapy.

[Malignant T-cell neoplasia during treatment with azathioprine and prednisone after kidney transplantation]. / Malign T-celle neoplasi under azatioprin- og prednisonbehandling efter nyretransplantation.

Patients receiving immunosuppressive treatment show an increased incidence of malignant lymphoproliferative disease, mainly in the form of B-lymphomata, on account of constant antigen stimulation from an implanted organ and/or impaired immunological defense which permit cells which have undergone mutation or have been transformed by viruses to proliferate. Malignant T-proliferative disease is rare in this patient category. The authors present a case in which a man treated with azathioprin and prednisone who had been well and without symptoms of rejection during a total of seven years after renal transplantation, which suggests that the T-cell neoplasia was due to the immunosuppressive treatment.

[Lactic acidosis associated with metformin]. / Metforminassocieret laktatacidose.

Lactic acidosis is an extremely rare but serious complication of treatment with metformin. In order to avoid lactate acidosis, it is important to select patients correctly and to ensure that contraindications such as cases of renal involvement, advanced age and chronic alcoholism are observed prior to treatment with metformin. In addition, it is necessary to carry out meticulous follow-up control of the patients with annual investigations of renal and hepatic functions and clinical assessment approximately four times per annum with the object of revealing commencing symptoms of diabetes and ischaemic conditions. In this manner, the majority of cases of lactate acidosis can be avoided.

[Laryngeal oncocytoid changes--hyperplasia or neoplasia?]. / Onkocytoide forandringer i larynx--hyperplasi eller neoplasi?

It is suggested that oncocytoid changes in glandular tissue represent a continuum in the development from metaplasia via increasing hyperplasia to neoplasia. This theory, which is supported in the literature, is based on three of the authors' own cases of oncocytoid changes in the larynx. The significance of differentiating between hyperplasia and neoplasia is emphasized as the neoplastic changes such as the oncocytoid papillary cystadenoma should be extirpated radically in order to avoid recurrence.

[Obstruction of the larynx caused by an oncocytic papillary cystadenoma]. / Larynxobstruktion forårsaget af et onkocytaert papillaert cystadenom.

A case of oncocytic papillary cystadenoma in the larynx in an elderly woman forms the basis for a discussion of the causes of, the symptoms and diagnosis of chronic obstructive conditions of the upper respiratory tract.