Sphingosylphosphorylcholine in Niemann-Pick Disease Brain: Accumulation in Type A But Not in Type B

A study of brain lipids in patients with the sphingomyelinase-deficient types of Niemann-Pick disease demonstrated that abnormal accumulation of sphingomyelin occurs only in the brain of neuronopathic type A patients but not in the non-neuronopathic type B. Additional lipid abnormalities were present in the type A brain. In contrast, the brain lipid profile was normal in type B patients. Since lysosphingolipids have been implicated in the biochemical pathogenesis of other genetic lysosomal sphingolipidoses, the occurrence of Sphingosylphosphorylcholine (lysosphingomyelin) was specifically investigated in brain and extraneural tissues, using an HPLC method with fluorescent detection of orthophtalaldehyde derivatives. Levels close to or below the limit of detection (10 pmol/mg tissue protein) were observed in normal and pathological controls. A striking accumulation was observed in brain of two Niemann-Pick type A patients (830 and 430 pmol/mg protein in 27-and 16-month-old children with severe and milder neurological course, respectively), which was not present at the fetal stage of the disease. No significant increase was found in brain tissue from a 3.5 year-old type B patient. In liver and spleen, abnormally high Sphingosylphosphorylcholine levels were observed in both types of the disease, with indication of a progressive increase during development. This study establishes the integrity of brain tissue in Niemann-Pick disease type B and suggests that the lysocompound Sphingosylphosphorylcholine could play a role in the pathophysiology of brain dysfunction in the neuronopathic type A.     

Are prognostic factors more favorable for breast cancer detected by organized screening than by opportunistic screening or clinical diagnosis? A study in Loire-Atlantique (France)

Purpose: Comparisons of breast cancer characteristics between organized and opportunistic screening have been limited. This study was designed to compare characteristics of cancers detected by either organized or opportunistic screening as well as clinically diagnosed cancers in Loire-Atlantique (a French administrative entity), from 2003 to 2007. Methods: This study is based on data from the population-based Loire-Atlantique Cancer Registry. Stage at diagnosis and prognostic characteristics of carcinomas detected by organized screening were compared, by age-adjusted logistic regressions, to those of cancers detected by opportunistic screening and diagnosed clinically. Analyses were restricted to women aged 50–74 years (the age group targeted by the organized screening program) for the 2003–2007 period. Results: Between 2003 and 2007, 2864 invasive and 400 in situ breast cancer cases were diagnosed in women aged 50–74 years in Loire-Atlantique. Compared to cancers diagnosed clinically, cancers detected by organized screening were more likely to be in situ (13.7% vs. 3.8%), diagnosed at an early stage (74.4% vs. 51.3%), have a low SBR grade (grade 1: 35.4% vs. 18.5%), and be positive for estrogen–progesterone receptors (68.3% vs. 59.0%). The distribution of stage at diagnosis and prognostic characteristics between organized and opportunistic screening were similar. Conclusion: These findings are consistent with the hypothesis that breast cancers are detected early by organized screening. Cancer characteristics were similar between the two screening modes. Estimating the impact of mammography screening on mortality in Loire-Atlantique should be the object of further investigations.     

The Niemann-Pick C1 protein resides in a vesicular compartment linked to retrograde transport of multiple lysosomal cargo

Niemann-Pick C disease (NP-C) is a neurovisceral lysosomal storage disorder. A variety of studies have highlighted defective sterol trafficking from lysosomes in NP-C cells. However, the heterogeneous nature of additional accumulating metabolites suggests that the cellular lesion may involve a more generalized block in retrograde lysosomal trafficking. Immunocytochemical studies in fibroblasts reveal that the NPC1 gene product resides in a novel set of lysosome-associated membrane protein-2 (LAMP2)(+)/mannose 6-phosphate receptor(-) vesicles that can be distinguished from cholesterol-enriched LAMP2(+) lysosomes. Drugs that block sterol transport out of lysosomes also redistribute NPC1 to cholesterol-laden lysosomes. Sterol relocation from lysosomes in cultured human fibroblasts can be blocked at 21 degrees C, consistent with vesicle-mediated transfer. These findings suggest that NPC1(+) vesicles may transiently interact with lysosomes to facilitate sterol relocation. Independent of defective sterol trafficking, NP-C fibroblasts are also deficient in vesicle-mediated clearance of endocytosed [14C]sucrose. Compartmental modeling of the observed [14C]sucrose clearance data targets the trafficking defect caused by mutations in NPC1 to an endocytic compartment proximal to lysosomes. Low density lipoprotein uptake by normal cells retards retrograde transport of [14C]sucrose through this same kinetic compartment, further suggesting that it may contain the sterol-sensing NPC1 protein. We conclude that a distinctive organelle containing NPC1 mediates retrograde lysosomal transport of endocytosed cargo that is not restricted to sterol.     

Catalytic Efficiency,Structure, and Recycling Behavior of Electrospun Polyvinyl Alcohol-Xylanase Fibers Cross-Linked by Glutaraldehyde

Food Biophysics - A versatile and effective method of producing polyvinyl alcohol (PVA)-xylanase (XY) fibers cross-linked by glutaraldehyde vapor (GA) is reported in the present study. Crosslinking...     

Isolation of NPC1-deficient Chinese hamster ovary cell mutants by gene trap mutagenesis

Chinese hamster ovary cell mutants defective in the NPC1 gene (NPC1-trap) were generated by retrovirus-mediated gene trap mutagenesis from a parental cell line JP17 expressing an ecotropic retrovirus receptor. Insertion of the gene trap vector in the NPC1 gene and the absence of the gene product were verified by 5'RACE and immunological analyses, respectively. NPC1-trap cells showed intracellular accumulation of low-density lipoprotein (LDL)-derived cholesterol and had an increased level of unesterified cellular cholesterol. Cholesterol biosynthesis through the mevalonate pathway was upregulated in the mutant cells as assessed by [(14)C]acetate incorporation into cellular sterols. When JP17 cells were depleted of lipoproteins and then loaded with LDL, cell surface LDL receptors were promptly downregulated and the mature form of the sterol regulatory element-binding protein-1 disappeared from the nucleus. These responses to LDL were obviously retarded in NPC1-trap cells, suggesting an impaired response of the cholesterol-regulatory system to LDL. NPC1-trap cells will be a useful tool to study the regulation of cellular cholesterol homeostasis and the pathogenesis of Niemann-Pick disease type C.