RÉSUMÉ
BACKGROUND: Autosomal recessive ABCA3 (ATP-binding cassette protein A3) gene mutations have been associated with neonatal respiratory distress and pediatric interstitial lung disease. The clinical course of the disease depends on the underlying mutations. Therefore, knowledge of course, symptoms and treatment of the disease is important. PATIENT AND METHODS: A term newborn suffered from progressive respiratory insufficiency, which led to death at the age of 4.8 months. The girl developed interstitial lung disease. Infections as well as structural and functional disorders of the lung were systematically excluded. A homozygous c.4681C > T (Arg 1561 Stop) mutation of the ABCA3 gene was identified. A literature review of the pathophysiology and treatment options of the disease was done. Therapeutic approaches with corticosteroids, macrolide, and hydroxychloroquine did not improve the clinical course. RESULTS: Therapeutic strategies for chronic interstitial lung disease have been used successfully in cases of a mild clinical course in juvenile patients with ABCA3 gene mutation. In our patient with homozygous ABCA3 gene mutation,they were not effective. Lung transplantation remains as a therapeutic option, but because of donor organ shortage and associated morbidity and mortality it is rarely feasible. CONCLUSION: More experience in the treatment of newborns with ABCA3 gene mutations is needed. Randomized, prospective evaluation of the different therapeutic approaches in a specific registry may improve prognosis and treatment of affected individuals.
Sujet(s)
Transporteurs ABC/génétique , Codon stop/génétique , Homozygote , Mutation/génétique , Syndrome de détresse respiratoire du nouveau-né/génétique , Insuffisance respiratoire/génétique , Hormones corticosurrénaliennes/usage thérapeutique , Aberrations des chromosomes , Issue fatale , Femelle , Gènes récessifs/génétique , Humains , Hydroxychloroquine/usage thérapeutique , Nourrisson , Nouveau-né , Pneumopathies interstitielles/traitement médicamenteux , Pneumopathies interstitielles/génétique , Macrolides/usage thérapeutique , Syndrome de détresse respiratoire du nouveau-né/traitement médicamenteux , Insuffisance respiratoire/traitement médicamenteux , Échec thérapeutiqueRÉSUMÉ
BACKGROUND AND AIM: The C1q/TNF-related protein (CTRP) family represents a growing family of adiponectin paralogous proteins. CTRP-5 was detected in adipose tissue of mice and plays a role in the context of the metabolic syndrome. It was the aim to investigate the detailed expression profile of CTRP-5 in a variety of adipocytic cells and to determine whether CTRP-5 circulates in human serum. Moreover, regulation and function of CTRP-5 was studied in the context of adipocyte biology. MATERIAL AND METHODS: CTRP-5 serum levels were measured in 50 healthy subjects by ELISA. Genotype analysis was performed by direct sequencing in 200 probands. CTRP-5 mRNA and protein expression was analyzed by RT-PCR, real-time RT-PCR and Western blot. Recombinant CTRP-5 and fatty acids were used for stimulation experiments in 3T3-L1 adipocytes. siRNA-mediated knockdown of CTRP-3 was performed during adipocyte differentiation. RESULTS: CTRP-5 mRNA and protein was strongly expressed in a wide variety of human and murine adipocytic cells and was induced during adipocyte differentiation. Saturated fatty acids increased CTRP-5 expression in adipocytes. siRNA-mediated cellular knockdown of CTRP-3 in adipocytes resulted in an upregulation of CTRP-5 expression. CTRP-5 inhibited the release of resistin and adiponectin dose-dependently. CTRP-5 circulates abundantly in human sera with a broad interindividual variation. The SNP 1014 T/A was not associated with type 2 diabetes mellitus in 200 Caucasian probands. CONCLUSIONS: CTRP-5 might be a novel adipokine that circulates abundantly in human sera. CTRP-5 is functionally involved in adipocyte biology and there might exist a counter-regulatory connection with its family member, CTRP-3.
Sujet(s)
Adipocytes/métabolisme , Adipocytes/physiologie , Protéines membranaires/génétique , Protéines membranaires/physiologie , Cellules 3T3-L1 , Adipocytes/effets des médicaments et des substances chimiques , Adipogenèse/effets des médicaments et des substances chimiques , Adipogenèse/génétique , Adulte , Animaux , Études cas-témoins , Cellules cultivées , Diabète de type 2/épidémiologie , Diabète de type 2/génétique , Diabète de type 2/anatomopathologie , Femelle , Régulation de l'expression des gènes/effets des médicaments et des substances chimiques , Régulation de l'expression des gènes/physiologie , Fréquence d'allèle , Humains , Mâle , Protéines membranaires/sang , Protéines membranaires/métabolisme , Souris , Souris de lignée C57BL , Adulte d'âge moyen , Polymorphisme de nucléotide simple/physiologie , Petit ARN interférent/pharmacologie , Jeune adulteRÉSUMÉ
The authors report, for the first time in the literature, a case of respiratory distress syndrome in a term baby due to homozygosity for a p.Trp308Arg/W308R substitution in the ATP-binding cassette transporter 3. The sequence was confirmed by genetic analysis of the baby and both parents. Management and long-term outcome of a patient carrying this novel genetic defect have not been reported in the literature before. Currently, lung transplant appears to be the only long-term survival option available, for which, our patient is being evaluated.
Sujet(s)
Transporteurs ABC/génétique , Mutation , Syndrome de détresse respiratoire du nouveau-né/génétique , Femelle , Homozygote , Humains , Nouveau-né , Naissance à termeRÉSUMÉ
The ATP-binding cassette transporter A1 (ABCA1) is a membrane-bound protein that is abundant in macrophages and is essential for the first step of reverse cholesterol transport and maintenance of homeostasis of high-density lipoprotein (HDL)-bound cholesterol. Low serum HDL levels are associated with increased risk for cardiovascular disease. Homozygous and heterozygous mutations in the ABCA1 gene may be associated with increased atherosclerosis. Here we report about two heterozygous mutations c.5398A>C and c.2369G>A in the ABCA1 gene associated with HDL cholesterol deficiency in serum.
Sujet(s)
Transporteurs ABC/génétique , Cholestérol HDL/génétique , Hypoalphalipoprotéinémies/génétique , Membre-1 de la sous-famille A des transporteurs à cassette liant l'ATP , Cholestérol HDL/sang , Humains , Adulte d'âge moyen , MutationRÉSUMÉ
Congenital insensitivity to pain with anhidrosis (CIPA) or hereditary sensory and autonomic neuropathy type IV is a rare, autosomal recessive neurologic disorder, characterized by absence of reaction to painful stimuli, mental retardation, self- mutilating behavior, anhidrosis, and recurrent episodes of hyperthermia. Mutations in the neurotrophic tyrosine kinase receptor 1, a receptor phosphorylated by nerve growth factor, have been documented in diverse ethnic groups. We identified the same novel nonsense mutation in two unrelated families of Moroccan Jewish descent, each with two affected siblings. This possible founder mutation may trace to the rural Jewish village in southern Morocco from where both these families originated. Genetic screening for the causative mutation among 300 unrelated Moroccan Jews did not reveal carriers for the causative mutation, thus excluding high risk for CIPA in this ethnic subpopulation.
Sujet(s)
Hypohidrose/génétique , Mutation , Analgésie congénitale/génétique , Récepteur trkA/génétique , Adolescent , Enfant , Famille , Femelle , Humains , Juif/génétique , Mâle , Maroc , Analgésie congénitale/ethnologie , PedigreeSujet(s)
Acné juvénile/génétique , Récepteur FGFR2/génétique , Récepteurs à l'interleukine-1/métabolisme , Tumeurs cutanées/génétique , Acné juvénile/anatomopathologie , Adolescent , Prolifération cellulaire , Cellules épithéliales , Humains , Mâle , Naevus/génétique , Naevus/anatomopathologie , Tumeurs cutanées/anatomopathologieRÉSUMÉ
INTRODUCTION: Cell culture media with high glucose concentration are normally used. Data on the secretion of the adipokines adiponectin and resistin from adipocytes in response to insulin and growth hormone (GH) both under normo- and hyperglycemic conditions are not available. It was the aim of the study to investigate the impact of standard metabolic conditions (normo-/hyperglycemia, normo-/hyperinsulinemia) and of GH on the secretion of adiponectin and resistin. MATERIAL AND METHODS: 3T3-L1 preadipocytes were differentiated into adipocytes and then incubated under normoglycemia (100 mg/dl), hyperglycemia (450 mg/dl), in combination with insulin (0, 0.2, 2.0 nM) and/or GH (1 nM). Adiponectin and resistin secretion was measured by ELISA. RESULTS: Insulin significantly stimulates adiponectin and resistin secretion under normo- and hyperglycemia. Hyperglycemia PER SE stimulates adiponectin and resistin secretion both in the absence and presence of low or high insulin concentrations. GH stimulates adiponectin secretion both under normoglycemic and hyperglycemic conditions. Whereas insulin does not modulate GH-induced adiponectin secretion under normoglycemia, insulin augments adiponectin release under hyperglycemia. GH stimulates resistin secretion only under normoglycemia, but not under hyperglycemic conditions. Since scavenger receptor B-I expression did not change, these effects are specific and not caused by a simple enhancement of adipocyte differentiation. DISCUSSION: Glucose, insulin and growth hormone have significant and interfering effects on the secretion of resistin and adiponectin. Several of the well-known in vivo phenomena such as diurnal variation or effects of re-feeding and weight-loss might be explained by direct effects of these hormones on adipocytes. Finally, when effects of hormones on adipocyte function are investigated, it is a prerequisite to take glucose levels of the cell culture media into account.
Sujet(s)
Adipocytes/effets des médicaments et des substances chimiques , Adipocytes/métabolisme , Adipokines/métabolisme , Glucose/administration et posologie , Hormone de croissance/pharmacologie , Insuline/pharmacologie , Cellules 3T3-L1 , Adipocytes/composition chimique , Adiponectine/métabolisme , Animaux , Technique de Western , Interactions médicamenteuses , Souris , Résistine/métabolisme , Récepteurs éboueurs de classe B/analyseRÉSUMÉ
Niemann-Pick type C disease is a fatal neurovisceral disorder linked to dysregulation in cholesterol processing. A medication for this disease is currently being tested in clinical trials. However, there is a lack of information on neuropsychological testing parameters for this disease. One aim of this pilot study was to evaluate a test battery that could be used to assess cognitive deficits in different stages of the disease. A second aim was to determine whether specific functional deficits are associated with certain disease stages. Eight men and two women (19-40 years of age) harbouring mutations in the gene coding for the cholesterol trafficking protein NPC1 were put through the same test battery independently of their disease stage. The external staging criterion was based on a five-step clinical scale. Trail Making tests A & B and verbal fluency were sensitive indicators at early stages of NPC. Corsi Block-Tapping, Mini Mental Status, Find Similarities and Clock Drawing showed abnormal results in patients with advanced disease. The Grooved Pegboard, Trail Making and Mosaic tests were unsuitable in advanced disease due to impaired fine motor skills. We observed that visuospatial working memory was less affected by the neurodegenerative process than verbal working memory. The series of tests used here could be supplemented by the severe impairment battery and Raven matrices tests for patients with advanced disease.
Sujet(s)
Protéines de transport/génétique , Glycoprotéines membranaires/génétique , Mutation , Maladies de Niemann-Pick/diagnostic , Maladies de Niemann-Pick/génétique , Maladies de Niemann-Pick/psychologie , Adulte , Études de cohortes , Évolution de la maladie , Femelle , Humains , Protéines et peptides de signalisation intracellulaire , Mâle , Mémoire , Aptitudes motrices , Tests neuropsychologiques , Protéine NPC1 , Facteurs tempsRÉSUMÉ
The aim of this controlled retrospective study was to evaluate the influence of an IL-1 gene polymorphism on the clinical and radiographic healing outcomes of GTR therapy. The study included 47 adult periodontitis patients with 94 deep intrabony defects treated by GTR using different membrane materials. The following clinical parameters were recorded at baseline and 12 months after surgery: papillary bleeding index (PBI), gingival recession (REC), probing pocket depth (PPD), clinical attachment level (CAL), and the vertical relative attachment gain (V-rAG). Bone changes in the defect regions due to GTR therapy were quantitatively evaluated using digital subtraction radiography (DSR). Polymorphisms of the IL-1A gene at position - 889 and of the IL-1B gene at position + 3953 were analyzed by PCR. Statistical analysis was performed using the Mann-Whitney-U and the Wilcoxon-Signed-Rank tests (alpha = 0.05). The study comprised 19 IL-1 genotype positive (IL-1 +) patients and 28 IL-1 genotype negative (IL-1 -) patients. Twelve months after GTR therapy, both patient groups revealed statistically significant PPD reductions and CAL gain [median (25/75% percentiles)]: Delta PPD [IL-1 + : 4.0 (2.5/5.0) mm; IL-1-: 3.8 (3.0/4.9) mm], Delta CAL [IL-1 + : 3.5 (3.0/4.8) mm; IL-1 -: 3.0 (1, 2/4, 5) mm]. V-rAG amounted to 60.0 (47.7/78.6)% in IL-1 + patients and 53.1 (43.4/81.9)% in IL-1 - patients. Both patient groups showed significant bone density gain in 40% (IL-1 +) and 43.6% (IL-1 -) of the initial defect area due to GTR. Neither the clinical nor the radiographic healing parameters revealed any statistically significant differences in the GTR healing outcome between IL-1 + and IL-1 - patients. In conclusion, these 12-month findings indicate that the IL-1 gene polymorphism has no influence on the clinical and radiographic regeneration results following GTR therapy.
Sujet(s)
Résorption alvéolaire/chirurgie , Régénération osseuse/génétique , Régénération tissulaire guidée parodontale , Interleukine-1/génétique , Polymorphisme génétique/génétique , Adulte , Résorption alvéolaire/imagerie diagnostique , Résorption alvéolaire/génétique , Densité osseuse/génétique , Femelle , Études de suivi , Hémorragie gingivale/chirurgie , Récession gingivale/chirurgie , Humains , Traitement d'image par ordinateur , Mâle , Adulte d'âge moyen , Perte d'attache parodontale/chirurgie , Poche parodontale/chirurgie , Parodontite/chirurgie , Radiographie , Études rétrospectives , Statistique non paramétrique , Technique de soustraction , Résultat thérapeutique , Cicatrisation de plaieRÉSUMÉ
Niemann-Pick disease type C (NPC) is an inherited neuro-degenerative disorder associated with intracellular cholesterol trafficking defects. Mutations in two distinct genes, NPC1 and HE1, have recently been shown to cause this disease. We have analysed the NPC1 gene in five German patients with NPC from four unrelated families. We identified a total of five novel mutations in the coding region of the NPC1 gene (G231V, D874V, 1642M, 11094T and R116stop). All affected individuals displayed compound heterozygosity. The mutated alleles were transmitted by the nonaffected parents with the exception of one patient, in whom a de novo mutation (G231V) had occurred. Interestingly, the G231V/P237S NPC1 genotype in this individual is associated with an early-onset form of NPC. In contrast, we found that the D874V/D948N genotype, observed in another NPC patient, is characterized by a late onset of clinical symptoms that presents with a pronounced white-matter disease. Our results will contribute to defining the association between the clinical phenotypes and the genetic abnormalities in Niemann-Pick C disease.
Sujet(s)
Mutation faux-sens , Maladies de Niemann-Pick/génétique , Adolescent , Adulte , Femelle , Allemagne , Hétérozygote , Humains , MâleRÉSUMÉ
BACKGROUND: The ABCA1 gene encodes for a member of subfamily A of the ATP-binding cassette transporters that plays an important role in cellular export of cholesterol and phospholipids; therefore, quantification of the ABCA1 mRNA is critical in many studies related to its expression and regulation by metabolic factors, nutritional status, and new antiatherogenic drug candidates. We developed a rapid, sensitive, specific, and reproducible real-time reverse transcription-PCR (RT-PCR) method for detection and quantification of ABCA1 transcripts in total RNA isolated from cultured human cells and tissues. METHODS: To quantify ABCA1 mRNA, we generated a calibration curve from serial dilutions of in vitro-transcribed RNA corresponding to an amplified ABCA1 cDNA 205-bp fragment (homologous calibrator). Two pairs of fluorescent hybridization probes were used to detect the ABCA1 and porphobilinogen deaminase (PBGD) mRNAs; the latter served as an internal control. PCR was performed as real-time amplification of ABCA1 mRNA in 100 ng of total RNA isolated from various human tissues, and cultured cells were calculated from the calibration curve. In addition, normalized values of target (ABCA1/PBGD ratio) were calculated. RESULTS: Using this method, we quantified ABCA1 transcripts in various human tissue samples as well as in monocytes, THP-1 cells, fibroblasts, and adipocytes. We demonstrated ABCA1 mRNA up-regulation during human adipocyte and monocyte differentiation. In addition, we examined the effect of cholesterol loading and deloading on ABCA1 expression in monocytes, THP-1 cells, and fibroblasts. CONCLUSIONS: Our RT-PCR assay allows the specific and highly reproducible detection and quantification of minute amounts of human ABCA1 mRNA. This new method is more accurate, more informative, and less laborious than the classic RT-PCR methods and Northern blot; it therefore could simplify all studies on ABCA1 mRNA expression.
Sujet(s)
Transporteurs ABC/génétique , ARN messager/analyse , Membre-1 de la sous-famille A des transporteurs à cassette liant l'ATP , Adipocytes/composition chimique , Technique de Northern , Cellules cultivées , Fibroblastes/composition chimique , Humains , Monocytes/composition chimique , Spécificité d'organe , Reproductibilité des résultats , RT-PCR , Sensibilité et spécificitéRÉSUMÉ
Familial high-density lipoprotein (HDL)-deficiency syndromes are caused by mutations of the ABCA1 gene, coding for the ATP-binding cassette transporter 1. We have developed a homogeneous assay based on 52 primer sets to amplify all 50 ABCA1 exons and approximately 1 kb of its promoter. The assay allows for convenient amplification of the gene from genomic DNA and easy mutational analysis through automatic sequencing. It obviates the need to use mRNA preparations, which were difficult to handle and posed a risk to miss splice junction or promoter mutations. The application of the test to the molecular analysis of a new patient with familial HDL-deficiency (Tangier disease) led to a discovery of two novel ABCA1 mutations: C2665del and C4457T.
Sujet(s)
Transporteurs ABC/génétique , Hypolipoprotéinémies/génétique , Lipoprotéines HDL/sang , Mutation , Membre-1 de la sous-famille A des transporteurs à cassette liant l'ATP , Adulte , Amorces ADN , Humains , Hypolipoprotéinémies/sang , Mâle , Pedigree , Réaction de polymérisation en chaîne/méthodes , SyndromeRÉSUMÉ
Pharmacogenomics deals with the interactions of individual genetic constitution with drug therapy. It has potentially far reaching consequences for drug development and future treatment strategies, but also for clinical in vitro diagnostics. With increasing knowledge about interactions between genes and drug treatment, there will be an equally increasing demand for rapid and reliable diagnostic tests prior to the institution of therapy. In fact, it is very likely that pharmacogenetic tests will make up a significant proportion of total molecular biology testing in the coming years. Therefore, this review focuses on the implications of pharmacogenomics on the clinical laboratory.
Sujet(s)
Techniques de laboratoire clinique/tendances , Pharmacogénétique/tendances , Conception de médicament , Techniques génétiques/tendances , Humains , Mutation/génétique , Polymorphisme génétique/génétique , Polymorphisme de nucléotide simple/génétique , Thérapeutique/tendancesRÉSUMÉ
HDL cholesterol (HDL-C) deficiency is the most common lipid abnormality observed in patients with premature coronary heart disease (CHD). Recently, our laboratory and others demonstrated that mutations in the ATP-binding cassette transporter 1 (ABCA1) gene are responsible for Tangier disease, a rare genetic disorder characterized by severely diminished plasma HDL-C concentrations and a predisposition for CHD. To address the question of whether common variants within the coding sequence of ABCA1 may affect plasma HDL-C levels and CHD risk in the general population, we determined the frequencies of three common ABCA1 variants (G596A, A2589G and G3456C) in men participating in the Veterans Affairs Cooperative HDL Cholesterol Intervention Trial (VA-HIT), a study designed to examine the benefits of HDL raising in men having low HDL-C (< or =40 mg/dl) and established CHD, as well as in CHD-free men from the Framingham Offspring Study (FOS). Allele frequencies (%) in VA-HIT were 31, 16, and 4 for the G596A, A2589G, and G3456C variants, respectively, versus 27, 12, and 2 in FOS (P<0.03). None of the variants were significantly associated with plasma HDL-C concentrations in either population; however, in VA-HIT, the G3456C variant was associated with a significantly increased risk for CHD end points, suggesting a role for this variant in the premature CHD observed in this population.
Sujet(s)
Transporteurs ABC/génétique , Cholestérol HDL/sang , Maladie coronarienne/sang , Maladie coronarienne/génétique , Variation génétique , Membre-1 de la sous-famille A des transporteurs à cassette liant l'ATP , Sujet âgé , Allèles , Fréquence d'allèle , Prédisposition génétique à une maladie , Humains , Mâle , Adulte d'âge moyenRÉSUMÉ
Congenital insensitivity to pain with anhidrosis (CIPA), also called hereditary sensory and autonomic neuropathy type IV (HSAN IV), is caused by mutations of the NTRK1 gene coding for the neurotrophic tyrosine kinase receptor type 1. We report the results of the NTRK1 sequence analysis in a CIPA family from Poland. We found that the patient was in a state of compound heterozygosity. He had one mutant allele with a novel G>A substitution in the conserved splice junction donor site affecting the first base pair of intron 5 (IVS5+1G>A). In the other allele he had a cluster of four single nucleotide substitutions in exon 15: an 1876C>T change (relative to the transcription start site) and three G>T changes (1904G>T, 1909G>T and 1915G>T). All of these mutations change the sense of the codons: H598Y, G607V, E609X and V611L, respectively. Mutations E609X and V611L are novel and unique to the patient family and at least one of them, which creates a premature stop codon in position 609, should have a deleterious effect on the gene function. The other two substitutions H598Y and G607V are most likely rare polymorphisms, which are in linkage disequilibrium. They occur together with an estimated allele frequency of about 6%. Our report increases the spectrum of NTRK1 mutations in CIPA patients and describes an unusual case of a cluster of four mutations located close to each other in one exon.
Sujet(s)
Allèles , Épissage alternatif/génétique , Exons/génétique , Neuropathies héréditaires sensitives et autonomes/génétique , Introns/génétique , Mutation/génétique , Récepteur trkA/génétique , Séquence d'acides aminés , Substitution d'acide aminé/génétique , Cellules cultivées , Enfant , Femelle , Dépistage des porteurs génétiques , Neuropathies héréditaires sensitives et autonomes/métabolisme , Humains , Mâle , Données de séquences moléculaires , PedigreeSujet(s)
Céroïdes-lipofuscinoses neuronales/génétique , Peptide hydrolases/génétique , Aminopeptidases , Dipeptidyl-Peptidases and Tripeptidyl-Peptidases , Endopeptidases , Femelle , Fluorescence , Marqueurs génétiques , Génotype , Humains , Mâle , Mutation , Réaction de polymérisation en chaîne , Protéases à sérine , Tripeptidyl-peptidase-1Sujet(s)
Analyse de mutations d'ADN/méthodes , Oxidoreductases acting on CH-NH group donors/génétique , Réaction de polymérisation en chaîne/méthodes , Prothrombine/génétique , Systèmes informatiques , Génome humain , Humains , Methylenetetrahydrofolate reductase (NADPH2) , Mutation ponctuelle , Spectrométrie de fluorescenceRÉSUMÉ
Tangier disease (TD) is an autosomal recessive disorder of lipid metabolism. It is characterized by absence of plasma high-density lipoprotein (HDL) and deposition of cholesteryl esters in the reticulo-endothelial system with splenomegaly and enlargement of tonsils and lymph nodes. Although low HDL cholesterol is associated with an increased risk for coronary artery disease, this condition is not consistently found in TD pedigrees. Metabolic studies in TD patients have revealed a rapid catabolism of HDL and its precursors. In contrast to normal mononuclear phagocytes (MNP), MNP from TD individuals degrade internalized HDL in unusual lysosomes, indicating a defect in cellular lipid metabolism. HDL-mediated cholesterol efflux and intracellular lipid trafficking and turnover are abnormal in TD fibroblasts, which have a reduced in vitro growth rate. The TD locus has been mapped to chromosome 9q31. Here we present evidence that TD is caused by mutations in ABC1, encoding a member of the ATP-binding cassette (ABC) transporter family, located on chromosome 9q22-31. We have analysed five kindreds with TD and identified seven different mutations, including three that are expected to impair the function of the gene product. The identification of ABC1 as the TD locus has implications for the understanding of cellular HDL metabolism and reverse cholesterol transport, and its association with premature cardiovascular disease.