Bull spermatozoa were examined for the presence and localization of constitutive Nitric Oxide Synthase (NOS), as nitric oxide (NO) is involved in calcium-dependent capacitation. In bull spermatozoa, NO generation is enhanced by l-arginine (3 microm) and abolished by the NOS-inhibitor N-nitro-l-arginine methyl ester (l-NAME). In addition, presence of NOS in bull spermatozoa was verified by immunohistochemistry, revealing the existence of both neuronal NOS (nNOS) and endothelial NOS (eNOS) immunoreaction. These findings were confirmed by Western blot technique, showing immunoreactive bands at 161 kDa (nNOS) and 133 kDa (eNOS). Confocal laser microscopy localized nNOS related immunofluorescence at the acrosome cap of sperms and their flagellum-mainpart. This technique also identified eNOS staining spread over the spermatozoan head. In conclusion, immunohistochemistry, Western blot technique, and NO generation suggest the presence of n- and eNOS in bull spermatozoa.
We demonstrated that the TT-genotype of the CD14 C(-260) T polymorphism in a South-German population is not associated with an increased risk of cerebral ischemia in general. However, we found that the TT-genotype is associated with a risk of atherosclerotic or microangiopathic stroke. This finding requires confirmation by future studies in larger populations.
Results of this study suggest a reference range that should be useful for aiding in the diagnosis of cyanide poisoning. Also, cattle can apparently accommodate a serum cyanide concentration of 206 microM without adverse effects.
Poisonings of domestic and wildlife animals are frequently caused by anticoagulants. Between 1996 and 2003, 70% of all suspected animal poisoning cases were confirmed in this laboratory following analysis of a variety of specimens. Thirty-nine percent of all animal poisonings detected in this laboratory were caused by anticoagulants. Suspicious veterinarians and pet owners involved with animal poisonings ask for toxicological analyses. With respect to these demands, a new high-performance liquid chromatography (HPLC) method (system-I, reversed-phase C-18 column, methanol/ammonium acetate) combined with specific extraction procedures was developed for the sensitive detection of nine substances with anticoagulative properties. This method yields high recoveries of the analytes, which can simultaneously be detected. In addition, another HPLC (system-II, acetonitrile/potassium phosphate) was used to confirm and separate chromatographic peaks, which could not be resolved by HPLC system-I. The reliable method has good reproducibility and is most suitable for forensic analyses. Using these analytical procedures, concentrations of eight organic anticoagulants were determined in more than 100 aliquots of animal liver, blood, bait, gastric content, bloody intestinal content, and water. Over the last eight years, dogs and wild birds were the most frequently poisoned species. Interestingly, lethally poisoned birds had lower anticoagulant residues in their organs than mammals.
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