ABSTRACT
The chromosome 9p21 amyotrophic lateral sclerosis-frontotemporal dementia (ALS-FTD) locus contains one of the last major unidentified autosomal-dominant genes underlying these common neurodegenerative diseases. We have previously shown that a founder haplotype, covering the MOBKL2b, IFNK, and C9ORF72 genes, is present in the majority of cases linked to this region. Here we show that there is a large hexanucleotide (GGGGCC) repeat expansion in the first intron of C9ORF72 on the affected haplotype. This repeat expansion segregates perfectly with disease in the Finnish population, underlying 46.0% of familial ALS and 21.1% of sporadic ALS in that population. Taken together with the D90A SOD1 mutation, 87% of familial ALS in Finland is now explained by a simple monogenic cause. The repeat expansion is also present in one-third of familial ALS cases of outbred European descent, making it the most common genetic cause of these fatal neurodegenerative diseases identified to date.
Subject(s)
Amyotrophic Lateral Sclerosis/genetics , Chromosomes, Human, Pair 9 , Frontotemporal Dementia/genetics , Microsatellite Repeats , Alleles , Female , Finland , Genetic Predisposition to Disease , Genotype , Haplotypes , Humans , Male , Pedigree , Polymorphism, Single NucleotideABSTRACT
The present study examined the impact of the menstrual cycle and oral contraceptive (OC) use on the growth hormone response to non-motorized treadmill sprinting. Nine monophasic OC users (21.5 ± 4.7 years old), and 8 normally menstruating women (NM; 21.4 ± 2.9 years old) participated in the study. Each participant completed 2 main trials, each consisting of an all-out 30-s treadmill sprint. The NM group performed one trial in the midfollicular phase (NM follicular) and one in the midluteal phase (NM luteal); the OC group's trials occurred one week into the start of the pill-taking cycle and once during the week in which pills were not taken.Venous blood samples were analyzed for growth hormone, pH, lactate, glucose, and progesterone concentrations. Peak and mean power output did not differ between the groups or with menstrual phase, or between the OC-free and OC trials. Integrated growth hormone was greater in the OC group than in the NM group (p = 0.04) with no phase difference (p = 0.80, mean (SD); NM follicular: 421 (335) and NM luteal: 345 (304) vs. OC free: 737 (471) and OC: 758 (389) µg·L(-1)·90 min(-1)). Blood lactate was higher in the OC group than in the NM group (p = 0.007) and, conversely, pH was lower in the OC group (p = 0.01). These results demonstrate that OC users who take high-androgenicity pills have a higher growth hormone response to sprint running than do normally menstruating women.
Subject(s)
Contraceptives, Oral/pharmacology , Human Growth Hormone/blood , Menstrual Cycle/physiology , Physical Exertion/physiology , Running/physiology , Adult , Blood Glucose , Contraceptives, Oral/blood , Female , Human Growth Hormone/drug effects , Humans , Hydrogen-Ion Concentration , Lactic Acid/blood , Progesterone/blood , Time Factors , Young AdultABSTRACT
BACKGROUND: Low density arrays (LDAs) have recently been introduced as a novel approach to gene expression profiling. Based on real time quantitative RT-PCR (QRT-PCR), these arrays enable a more focused and sensitive approach to the study of gene expression than gene chips, while offering higher throughput than more established approaches to QRT-PCR. We have now evaluated LDAs as a means of determining the expression of multiple genes simultaneously in human tissues and cells. RESULTS: Comparisons between LDAs reveal low variability, with correlation coefficients close to 1. By performing 2-fold and 10-fold serial dilutions of cDNA samples in the LDAs we determined a clear linear relationship between the gene expression data points over 5 orders of magnitude. We also showed that it is possible to use LDAs to accurately and quantitatively detect 2-fold changes in target copy number as well as measuring genes that are expressed with low and high copy numbers in the range of 1 x 10(2)-1 x 10(6) copies. Furthermore, the data generated by the LDA from a cell based pharmacological study were comparable to data generated by conventional QRT-PCR. CONCLUSION: LDAs represent a valuable new approach for sensitive and quantitative gene expression profiling.
Subject(s)
Gene Expression Profiling/methods , Oligonucleotide Array Sequence Analysis/methods , DNA Primers , Endothelium, Vascular/cytology , Endothelium, Vascular/drug effects , Endothelium, Vascular/metabolism , Humans , RNA, Messenger/analysis , RNA, Messenger/metabolism , Reproducibility of Results , Reverse Transcriptase Polymerase Chain Reaction/methods , Thrombin/pharmacologyABSTRACT
Increased release of oxidants has been implicated in the pathogenesis of pulmonary fibrosis. Previous work in the rat showed that formation of the early fibrotic lesion is associated with increased expression of inducible nitric oxide synthase (iNOS) in pulmonary fibroblasts. The aim of this study was to test the hypothesis that NO is involved in the activation of pulmonary fibroblasts. The effects of endogenous and exogenous NO on proliferation of human pulmonary fibroblasts were investigated by administration of cytomix or SNAP, respectively. At low concentrations, both treatments increased cell numbers, an effect attenuated by iNOS inhibitor or NO scavenger. Induction of iNOS was confirmed by measurement of nitrate/nitrite production and by immunodetection. Quantitative RT-PCR showed an increase in iNOS mRNA as early as 3 h after stimulation. These results support the hypothesis and show that upregulation of the iNOS gene is an early event in the proliferative response of human lung fibroblasts to inflammatory stimuli.
