Assessment of plasma chitotriosidase activity, CCL18/PARC concentration and NP-C suspicion index in the diagnosis of Niemann-Pick disease type C: A prospective observational study

Background: Niemann-Pick disease type C (NP-C) is a rare, autosomal recessive neurodegenerative disease caused by mutations in either the NPC1 or NPC2 genes. The diagnosis of NP-C remains challenging due to the non-specific, heterogeneous nature of signs/symptoms. This study assessed the utility of plasma chitotriosidase (ChT) and Chemokine (C-C motif) ligand 18 (CCL18)/pulmonary and activation-regulated chemokine (PARC) in conjunction with the NP-C suspicion index (NP-C SI) for guiding confirmatory laboratory testing in patients with suspected NP-C. Methods: In a prospective observational cohort study, incorporating a retrospective determination of NP-C SI scores, two different diagnostic approaches were applied in two separate groups of unrelated patients from 51 Spanish medical centers (n = 118 in both groups). From Jan 2010 to Apr 2012 (Period 1), patients with =2 clinical signs/symptoms of NP-C were considered ''suspected NP-C'' cases, and NPC1/NPC2 sequencing, plasma chitotriosidase (ChT), CCL18/PARC and sphingomyelinase levels were assessed. Based on findings in Period 1, plasma ChT and CCL18/PARC, and NP-C SI prediction scores were determined in a second group of patients between May 2012 and Apr 2014 (Period 2), and NPC1 and NPC2 were sequenced only in those with elevated ChT and/or elevated CCL18/PARC and/or NP-C SI =70. Filipin staining and 7-ketocholesterol (7-KC) measurements were performed in all patients with NP-C gene mutations, where possible. Results: In total across Periods 1 and 2, 10/236 (4%) patients had a confirmed diagnosis o NP-C based on gene sequencing (5/118 4.2%] in each Period): all of these patients had two causal NPC1 mutations. Single mutant NPC1 alleles were detected in 8/236 (3%) patients, overall. Positive filipin staining results comprised three classical and five variant biochemical phenotypes. No NPC2 mutations were detected. All patients with NPC1 mutations had high ChT activity, high CCL18/PARC concentrations and/or NP-C SI scores =70. Plasma 7-KC was higher than control cut-off values in all patients with two NPC1 mutations, and in the majority of patients with single mutations. Family studies identified three further NP-C patients. Conclusion: This approach may be very useful for laboratories that do not have mass spectrometry facilities and therefore, they cannot use other NP-C biomarkers for diagnosis

Antioxidant activity and duodenum transporter gene expression in quail fed citric acid

The study aimed to evaluate the effects of citric acid supplementation on the performance and the gene expression in the duodenum of Japanese quail. Genes related to antioxidant activity: superoxide dismutase (SOD) and glutathione peroxidase 7 (GPX7); and genes related to nutrient uptake in the enterocytes’ apical membrane: sodium-glucose cotransporter 1 (SGLT1, responsible for the glucose absorption), glucose transporter 2 (GLUT2) and B0AT1 (responsible for the absorption of neutral amino acids in brush board membrane) were evaluated. For this, 270 laying quails (Coturnix japonica) were fed a diet supplemented with 0%, 0.6% or 1.2% of citric acid. Gene expression was evaluated by the quantitative reverse transcription-polymerase chain reaction (qRT-PCR) at 35 days old (n = 5). The highest expression of the SGLT1 gene was observed in quails fed the 1.2% citric acid supplemented diet (0.120 AU). Birds fed diets with both levels of citric acid supplementation showed a higher B0AT1 gene expression than quails fed no citric acid. Quails fed the 1.2% of citric acid supplemented diet had the lowest SOD gene expression (0.3455 AU). The birds receiving the diets supplemented with citric acid showed a lower GPX7 gene expression than the quails fed the 0% citric acid diet. These results suggests that the effect of citric acid on gene expression in the digestive tract may have contributed to the greater weight gain and lower feed intake observed in birds fed diets supplemented with citric acid.Keywords: B0AT1, intestinal health, organic acid, pathogenic microorganisms, SGLT

Characterization of Isomeric Cationic Porphyrins with β-Pyrrolic Substituents by Electrospray Mass Spectrometry: The Singular Behavior of a Potential Virus Photoinactivator

Electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (ESI-MS/MS) have been used to differentiate the 2- and 4-methylpyridyl isomers of free-base and metallated cationic β-vinylpyridylporphyrins. The analysis by ESI-MS/MS of the deuterated analogs and semiempirical calculations of structural and electronic parameters were also undertaken. The two free-base isomers are easily differentiated by ESI-MS/MS but the presence of a metallic center renders differentiation of the metallated isomers less effective. The data acquired show that of all the studied compounds, the free-base 2-methylpyridyl isomer, which was operative in the in vitro photoinactivation of Herpes simples virus, has a different gas-phase behavior. Local distortion of the macrocycle due to the presence of the β-vinylpyridyl substituent occurs for all the compounds, but a different electron density distribution can account for the observed gas-phase behavior of this potential virus photoinactivator