Novel Pathogenic PRSS1 Variant p.Glu190Lys in a Case of Chronic Pancreatitis

Mutations in the PRSS1 (serine protease 1) gene encoding human cationic trypsinogen cause hereditary pancreatitis or may be associated with sporadic chronic pancreatitis. The mutations exert their pathogenic effect either by increasing intra-pancreatic trypsinogen activation (trypsin pathway) or by causing proenzyme misfolding and endoplasmic reticulum stress (misfolding pathway). Here we report a novel heterozygous c.568G>A (p.Glu190Lys) variant identified in a case with chronic pancreatitis. The parents of the index patient had no history of pancreatitis but were unavailable for genetic testing. Functional characterization revealed 2.5-fold increased autoactivation of the mutant trypsinogen relative to wild type. Unlike many other clinically relevant PRSS1 mutations, p.Glu190Lys did not alter the chymotrypsin C (CTRC)-dependent degradation of trypsinogen nor did it increase CTRC-mediated processing of the trypsinogen activation peptide. Cellular secretion of the mutant protein was unchanged indicating normal folding behavior. Based on the genetic and functional evidence, we classify the p.Glu190Lys PRSS1 variant as likely pathogenic, which stimulates autoactivation of cationic trypsinogen independently of CTRC

Electrophysiological and biochemical response in rats on intratracheal instillation of manganese

Abstract Chronic exposure to excess manganese via inhalation of metal fumes causes central nervous system damage. For modelling Mn aerosol inhalation, male Wistar rats were intratracheally instilled with MnCl2 solution (0.5 mg/kg b.w. MnCl2; n=12) 5 days a week for 5 weeks. At the end of the treatment, somatosensory cortical evoked potentials, elicited by double-pulse stimulation, were recorded from the animals in urethane anaesthesia. Body weight gain, organ weights, and Mn level in brain, lung and blood samples were also measured. In brain samples, gene expression level of MnSOD (Mn superoxide dismutase) was determined. The effect of Mn was mainly seen on the evoked potential amplitudes, and on the second:first ratio of these. Tissue Mn concentration was elevated in brain and lungs, but changed hardly in the blood. Relative weight of heart, thymus, lungs and brain was significantly altered. The level of MnSOD transcript in brain tissue decreased. The observed effects showed that Mn had access to the brain and that somatosensory cortical responses evoked by double-pulse stimulation might be suitable biomarkers of Mn intoxication.</jats:p

Gut region-specific rearrangement of the cellular and subcellular compartments of nitric oxide synthase isoforms after chronic ethanol consumption in rats

We recently provided evidence of cell-typespecific differences in the subcellular distributions of the three nitric oxide synthase (NOS) isoforms in the myenteric neurons, enteric smooth muscle cells and the capillary endothelium of the rat duodenum. We hypothesized that the presence of three NOS isoforms in the same type of cells with differences in subcellular compartmentalization might reflect a functional plasticity. Therefore, investigation of the possible rearrangement of cellular and subcellular NOS compartments in different gut segments following chronic ethanol treatment was the aim of this study. Rats were randomly divided into two groups and received water or 20% ethanol solution, preceded by short periods of adaptation with 10% and 15% ethanol. After 8 weeks, segments of duodenum, ileum and colon of the control and the alcohol-treated rats were processed for post-embedding immunohistochemistry and RT-PCR. The quantitative differences in the numbers of gold particles indicative of the different NOSs and their relative mRNA levels between the two experimental groups varied greatly, depending on the gut segment, and also on the cellular and subcellular compartments investigated. The chronic ethanol administration had the opposite effect on the quantitative distribution of the neuronal and endothelial NOS labelling gold particles in the different cellular compartments and resulted in subcellular rearrangement of NOS labels along the gastrointestinal tract. The intestinal region-specific rearrangement of the cellular and subcellular NOS compartments may possibly result in functional plasticity and help to maintain the optimum NO level under pathological conditions