ABSTRACT
A repeat expansion mutation in the C9orf72 gene is the leading known genetic cause of FTD and ALS. The C9orf72-ALS/FTD field has been plagued by a lack of reliable tools to monitor this genomic locus and its RNA and protein products. We have validated assays that quantify C9orf72 pathobiology at the DNA, RNA and protein levels using knock-out human iPSC lines as controls. Here we show that single-molecule sequencing can accurately measure the repeat expansion and faithfully report on changes to the C9orf72 locus in what has been a traditionally hard to sequence genomic region. This is of particular value to sizing and phasing the repeat expansion and determining changes to the gene locus after gene editing. We developed ddPCR assays to quantify two major C9orf72 transcript variants, which we validated by selective excision of their distinct transcriptional start sites. Using validated knock-out human iPSC lines, we validated 4 commercially available antibodies (of 9 tested) that were specific for C9orf72 protein quantification by Western blot, but none were specific for immunocytochemistry. We tested 15 combinations of antibodies against dipeptide repeat proteins (DPRs) across 66 concentrations using MSD immunoassay, and found two (against poly-GA and poly-GP) that yielded a 1.5-fold or greater signal increase in patient iPSC-motor neurons compared to knock-out control, and validated them in human postmortem and transgenic mouse brain tissue. Our validated DNA, RNA and protein assays are applicable to discovery research as well as clinical trials.
Subject(s)
Amyotrophic Lateral Sclerosis , Craniocerebral Trauma , Frontotemporal Dementia , Animals , Mice , Humans , Amyotrophic Lateral Sclerosis/genetics , C9orf72 Protein/genetics , Antibodies , Mice, Transgenic , DNA , RNAABSTRACT
OBJECTIVE: To determine the accuracy of 1.5 Tesla magnetic resonance imaging in identifying the bony defect in superior semi-circular canal dehiscence syndrome. METHODS: A retrospective case-control study was conducted in tertiary university settings. The study included 12 patients with a definite diagnosis of superior semi-circular canal dehiscence syndrome and a control group comprising 12 non-superior semi-circular canal dehiscence syndrome patients, all with available magnetic resonance imaging data. The imaging scans were anonymised, and reviewed blindly and independently by three experienced radiologists. RESULTS: There was low sensitivity and specificity, with average values of 47 per cent (range, 16.7-66.7 per cent) and 69 per cent (range, 66.7-75 per cent) respectively. Cohen's kappa was 0.75, indicating substantial inter-rater reliability. CONCLUSION: Given the low accuracy of 1.5T magnetic resonance imaging in identifying the bony defect in superior semi-circular canal dehiscence syndrome, despite its value in inner-ear imaging, computed tomography or equivalent should be used when superior semi-circular canal dehiscence syndrome is suspected.
Subject(s)
Magnetic Resonance Imaging , Semicircular Canals , Humans , Case-Control Studies , Magnetic Resonance Imaging/methods , Reproducibility of Results , Retrospective Studies , Semicircular Canals/diagnostic imaging , Semicircular Canals/pathologyABSTRACT
Ectopic submandibular thyroid tissue is a rare entity and poses difficult diagnostic and management problems. The first case of ectopic submandibular thyroid with a normotopic multinodular goitre is presented
Subject(s)
Choristoma/diagnosis , Goiter, Nodular/diagnosis , Thyroid Gland , Aged , Biopsy, Fine-Needle , Choristoma/complications , Choristoma/surgery , Female , Goiter, Nodular/complications , Goiter, Nodular/surgery , Humans , Neck/diagnostic imaging , Thyroidectomy , Tomography, X-Ray ComputedABSTRACT
Nicotine dependence plays a critical role in addiction to tobacco products, and thus contributes to a variety of devastating tobacco-related diseases (SGR 2014). Annual costs associated with smoking in the US are estimated to be between $289 and $333 billion. Effective interventions for nicotine dependence, especially in smokers, are a critical barrier to the eradication of tobacco-related diseases. This overview highlights research presented at the Plenary Symposium of Behavior, Biology and Chemistry: Translational Research in Addiction Conference (BBC), hosted by the UT Health Science Center San Antonio, on March 9-10, 2013. The Plenary Symposium focused on tobacco addiction, and covered topics ranging from basic science to national policy. As in previous years, the meeting brought together globally-renowned scientists, graduate student recruits, and young scientists from underrepresented populations in Texas and other states with the goal of fostering interest in drug addiction research in young generations.
Subject(s)
Behavior, Addictive/drug therapy , Smoking Cessation/methods , Smoking/drug therapy , Substance Withdrawal Syndrome/diagnosis , Tobacco Use Disorder/drug therapy , Behavior, Addictive/physiopathology , Humans , Research , Smoking/physiopathology , Substance Withdrawal Syndrome/physiopathology , Tobacco Use Disorder/physiopathologySubject(s)
Fibrinopeptide A/immunology , Thrombosis/diagnosis , Animals , Biomarkers , Cross Reactions , HumansABSTRACT
Real-time PCR methods with primers and a probe targeting conserved regions of the bacterial 16S ribosomal DNA (rDNA) revealed a larger amount of rDNA in blood specimens from healthy individuals than in matched reagent controls. However, the origins and identities of these blood-associated bacterial rDNA sequences remain obscure.
Subject(s)
DNA, Bacterial/blood , DNA, Ribosomal/blood , Polymerase Chain Reaction/methods , RNA, Ribosomal, 16S/genetics , Bacteria/genetics , DNA Primers/genetics , HumansABSTRACT
Urines from 19 autistic children (3 female, 16 male) were analyzed by high-resolution, high-pressure anion-exchange chromatography. The results showed abnormalities in the excretion of hippuric acid, 4-hydroxyhippuric acid, and N1-methyl-2-pyridone-5-carboxamide, the end product of the nicotinic acid pathway. Considerations as the metabolic origin on the 4-hydroxyhippuric acid are discussed and related to the possibility of bacterial action and malabsorption.
Subject(s)
Autistic Disorder/urine , Hippurates/urine , Autistic Disorder/drug therapy , Bacteria/metabolism , Child , Chromatography, High Pressure Liquid , Chromatography, Ion Exchange , Digestive System/microbiology , Female , Humans , Malabsorption Syndromes/urine , Male , Nicotinic Acids/metabolism , Spectrophotometry, Ultraviolet , Toxins, Biological/biosynthesis , Vitamins/therapeutic useABSTRACT
The ultrasonic features of 162 bladder tumours are described. Comparing clinical staging by the TNM system and the ultrasonic appearances, there is a high degree of accuracy in staging by ultrasound. The use of this painless non-invasive technique is assessed and should be used in staging bladder tumours only in conjunction with other established methods, and not in isolation.
Subject(s)
Ultrasonography , Urinary Bladder Neoplasms/pathology , Cystoscopy , Humans , Infant, Newborn , Obesity/complications , Radiography , Urinary Bladder Neoplasms/diagnosis , Urinary Bladder Neoplasms/diagnostic imagingABSTRACT
Thirty-three patients with end-stage renal failure have had transplants over a three-year period, four patients receiving kidneys from siblings and the remainder cadaver organs. Twenty-seven kidneys survived with stable function for periods of six months to three years. Graft survival at one year was 85% and at two years 82%. One patient died and five were returned to dialysis. Complications included rejection episodes, technical problems, respiratory and wound infections, gastrointestinal disorders, and side effects of steroids.