Crustaceans can store excess copper in the hepatopancreas, an organ playing a role in digestive activity as well as in neurosecretory control. Here, we studied the effect of copper exposure on the level of histamine, an indicator of food spoilage in edible crustaceans. Histamine is also a neuromodulator in the intestinal nervous system of crustaceans, and a human allergen. Marbled crayfish {Procambarus fallax forma virginalis) were exposed to average measured values of 0.031 mg Cu/1 and 0.38 mg Cu/1, respectively, for 14 days and then transferred to copper-free water for another 14 days. Concentrations of copper and histamine in the hepatopancreas and muscle were evaluated at different time points. Histamine levels were significantly higher in hepatopancreas and muscle tissues at the highest exposure level, but only after transfer of the animals to copper-free water. The increased histamine concentration following copper exposure may be explained by a (delayed) stress response, and by up-regulated histidine synthesis induced by copper, followed by decarboxylation to histamine.
In patients with high-grade squamous intraepithelial lesion (HSIL) of the vulva, the presence of multiple lesions, called multifocal HSIL, is common. The aim of this exploratory study was to investigate biomarker expression profiles in multifocal HSIL. In total, 27 lesions from 12 patients with high-risk human papillomavirus (HPV)-positive multifocal HSIL were tested for HPV genotype, expression of p16INK4a and Ki-67, and DNA methylation of six genes. HPV16 was found most commonly in 21 (77.8%) HSILs. In two (16.4%) patients, HPV genotype differed between the lesions. All lesions demonstrated diffuse p16INK4a staining, of which three (11.1%) were combined with patchy staining. One patient (8.3%) demonstrated markedly different DNA methylation levels between lesions. Generally, heterogeneity in methylation profiles was observed between different patients, even when other biomarkers showed similar expression. In conclusion, this study is the first to demonstrate heterogeneity of individual lesions in patients with multifocal HSIL. The studied biomarkers have the potential to refine prognostic and predictive diagnostics. Future prospective, longitudinal studies are needed to further explore the potential of a biomarker profile for management of patients with multifocal HSIL.
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