Polar bear encephalitis: establishment of a comprehensive next-generation pathogen analysis pipeline for captive and free-living wildlife.

This report describes three possibly related incidences of encephalitis, two of them lethal, in captive polar bears (Ursus maritimus). Standard diagnostic methods failed to identify pathogens in any of these cases. A comprehensive, three-stage diagnostic 'pipeline' employing both standard serological methods and new DNA microarray and next generation sequencing-based diagnostics was developed, in part as a consequence of this initial failure. This pipeline approach illustrates the strengths, weaknesses and limitations of these tools in determining pathogen caused deaths in non-model organisms such as wildlife species and why the use of a limited number of diagnostic tools may fail to uncover important wildlife pathogens.

Plasmin-induced expression of cytokines and tissue factor in human monocytes involves AP-1 and IKKbeta-mediated NF-kappaB activation.

It was previously shown that plasmin activates human peripheral monocytes in terms of lipid mediator release and chemotactic migration. Here it is demonstrated that plasmin induces proinflammatory cytokine release and tissue factor (TF) expression by monocytes. Plasmin 0.043 to 1.43 CTA U/mL, but not active site-blocked plasmin, triggered concentration-dependent expression of mRNA for interleukin-1alpha (IL-1alpha), IL-1beta, tumor necrosis factor-alpha (TNF-alpha), and TF with maximum responses after 4 hours. Plasmin-mediated mRNA expression was inhibited in a concentration-dependent manner by the lysine analogue trans-4-(aminomethyl)cyclohexane-1-carboxylic acid (t-AMCA). Increases in mRNA levels were followed by concentration- and time-dependent release of IL-1alpha, IL-1beta and TNF-alpha and by TF expression on monocyte surfaces. Neither cytokines nor TF could be detected when monocytes were preincubated with actinomycin D or cycloheximide. Electrophoretic mobility shift assays indicated plasmin-induced activation of NF-kappaB; DNA-binding complexes were composed of p50, p65, and c-Rel, as shown by supershift experiments. Nuclear translocation of NF-kappaB/Rel proteins coincided with IkappaBalpha degradation. At variance with endotoxic lipopolysaccharide, plasmin elicited the rapid degradation of another cytoplasmic NF-kappaB inhibitor, p105. Proteolysis of NF-kappaB inhibitors was apparently due to transient activation of IkappaB kinase (IKK) beta that reached maximum activity at 1 hour after plasmin stimulation. In addition, AP-1 binding was increased in plasmin-treated monocytes, with most complexes composed of JunD, c-Fos, and FosB. These findings further substantiate the role of plasmin as a proinflammatory activator of human monocytes and reveal an important new link between the plasminogen-plasmin system and inflammation. (Blood. 2001;97:3941-3950)

Cardiac effects of lappaconitine and N-deacetyllappaconitine, two diterpenoid alkaloids from plants of the Aconitum and Delphinium species.

Aconitum and Delphinium alkaloids are currently under investigation in search for new analgesic and anti-inflammatory drugs. It has been reported that the analgesic compound lappaconitine (LA), a C19 diterpenoid alkaloid from Aconitum sinomontanum nakai is an inhibitor of tetrodotoxin-sensitive, voltage-dependent sodium channels. In the present study we investigated the cardiac effects of LA and its metabolite N-deacetyllappaconitine (DLA) in electrically stimulated left and spontaneously beating right atria isolated from guinea-pig hearts. In all experiments, equieffective concentrations were larger with DLA than with LA. At a stimulation frequency of 2.5 Hz the time constant for the onset of LA effects (tau = 56 +/- 29 min) was markedly larger than the one for DLA effects (tau = 14 +/- 8 min). The compounds exerted a significant negative inotropic action at 0.06 microM (LA) and 0.2 microM (DLA). Asystolia of right atria occurred at 4.5 microM (LA) and 10 microM (DLA). Therefore, cardiotoxicity of LA and DLA was much lower compared to aconitine, which caused arrhythmia at 10 nM in our model. For both alkaloids a use-dependent mode of action could be demonstrated. In addition, preincubation with 0.3 microM LA prevented arrhythmia induced by aconitine or ouabain. We conclude that lappaconitine is a naturally occurring compound with class-I antiarrhythmic action.

Risk factors for acquired myasthenia gravis in dogs: 1,154 cases (1991-1995).

OBJECTIVE: To determine frequency of initial clinical signs and risk factors for acquired myasthenia gravis (MG) in dogs. DESIGN: Retrospective study. SAMPLE POPULATION: 1,154 dogs residing within the United States from 1991 to 1995 with a confirmed diagnosis of acquired MG and 7,176 dogs with other neuromuscular disorders, including generalized weakness, megaesophagus, and dysphagia (control group). PROCEDURE: Records were retrieved from a database containing results of serum samples tested for acetylcholine receptor antibodies. Signalment, breed, age, state of origin, and month of onset of clinical signs were obtained. An antibody titer > 0.6 nmol/L was diagnostic for acquired MG. Unconditional logistic regression was used for statistical analysis. RESULTS: In comparison with mixed-breed dogs, dogs with the highest risk of acquired MG were Akitas, terrier group, Scottish Terriers, German Shorthaired Pointers, and Chihuahuas. Rottweilers, Doberman Pinschers, Dalmatians, and Jack Russell Terriers had low relative risks. Sexually intact males and dogs less than 1 year old had some protection from risk. Generalized weakness with megaesophagus and megaesophagus alone were the most common initial clinical signs. CLINICAL IMPLICATIONS: Breed predispositions for acquired MG were demonstrated. Age and sex were contributing factors. Although most dogs had generalized clinical signs, a substantial proportion of dogs had focal signs.

[3H]-monoamine uptake inhibition properties of kava pyrones.

Three kava pyrones, the natural compounds (+)-methysticine and (+)-kavain, and the synthetic racemate (+/-)-kavain, were tested concerning their action on in vitro uptake of monoamines in synaptosomes prepared from the cerebral cortex and hippocampus of rats. (+/-)-Kavain and (+)-kavain were found to potently inhibit the uptake of [3H]-noradrenaline. Uptake of [3H]-noradrenaline was inhibited in the following order of potency: (+/-)-kavain = (+)-kavain > (+)-methysticine, whereas none of the kava pyrones efficiently blocked the uptake of [3H]-serotonin. The results indicate a pyrone-specific non-stereo-selective inhibition of the [3H]-noradrenaline uptake which might be responsible for or, at least, contribute to the psychotropic properties of kava pyrones.

[Are loss of glistening of normal colour and increased friability normal aspects of the oesophagus in old age? (author's transl)]. / Glanzverlust, Farbveränderungen und erhöhte Lädierbarkeit der Speiseröhre: altersbedingte Normvarianten?

Friability of the esophageal mucosq increases with old age. Old patients without esophageal disease also show loss of glistening and of the normal pink color of the mucosa. These findings on their own are therefore no signs of esophagitis,

[Endoscopic diagnosis of oesophagitis: problems of differentiation from normal (author's transl)]. / Endoskopische Diagnose der Osophagitis Wo liegt die Grenze zum Normalen

The diagnostic significance of endoscopically early signs of oesophagitis in the absence of macroscopic evidence of epithelial abnormalities has been studied prospectively. Changes from the usually observed shininess, pale-pink colour, smooth texture, regular capillaries, sharp Z-line and a decreased mechanical resistance of the mucosa to mechanical damage do not indicate oesophagitis. Nor should one infer gastro-oesophageal reflux from these endoscopic findings. They are presumably ageing changes in the oesophagus.