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1.
BMC Neurol ; 24(1): 31, 2024 Jan 17.
Artigo em Inglês | MEDLINE | ID: mdl-38233770

RESUMO

BACKGROUND: SCN8A-related disorders are a group of variable conditions caused by pathogenic variations in SCN8A. Online Mendelian Inheritance in Man (OMIM) terms them as developmental and epileptic encephalopathy 13, benign familial infantile seizures 5 or cognitive impairment with or without cerebellar ataxia. METHODS: In this study, we describe clinical and genetic results on eight individuals from six families with SCN8A pathogenic variants identified via exome sequencing. RESULTS: Clinical findings ranged from normal development with well-controlled epilepsy to significant developmental delay with treatment-resistant epilepsy. Three novel and three reported variants were observed in SCN8A. Electrophysiological analysis in transfected cells revealed a loss-of-function variant in Patient 4. CONCLUSIONS: This work expands the clinical and genotypic spectrum of SCN8A-related disorders and provides electrophysiological results on a novel loss-of-function SCN8A variant.


Assuntos
Disfunção Cognitiva , Epilepsia Generalizada , Epilepsia , Humanos , Epilepsia/genética , Genótipo , Fenótipo , Mutação/genética , Canal de Sódio Disparado por Voltagem NAV1.6/genética
2.
Bioorg Med Chem Lett ; 45: 128133, 2021 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-34044121

RESUMO

We describe the synthesis and biological evaluation of a series of novel aryl sulfonamides that exhibit potent inhibition of NaV1.5. Unlike local anesthetics that are currently used for treatment of Long QT Syndrome 3 (LQT-3), the most potent compound (-)-6 in this series shows high selectivity over hERG and other cardiac ion channels and has a low brain to plasma ratio to minimize CNS side effects. Compound (-)-6 is also effective inshortening prolonged action potential durations (APDs) in a pharmacological model of LQT-3 syndrome in pluripotent stem cell-derived cardiomyocytes (iPSC-CMs). Unlike most aryl sulfonamide NaV inhibitors that bind to the channel voltage sensors, these NaV1.5 inhibitors bind to the local anesthetic binding site in the central pore of the channel.


Assuntos
Canal de Sódio Disparado por Voltagem NAV1.5/metabolismo , Sulfonamidas/farmacologia , Relação Dose-Resposta a Droga , Humanos , Estrutura Molecular , Relação Estrutura-Atividade , Sulfonamidas/síntese química , Sulfonamidas/química
3.
Clin Genet ; 85(3): 282-5, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23509941

RESUMO

Individuals with mosaic paternal uniparental disomy (UPD) of apparently all chromosomes have recently been described. They show a 46,XX karyotype, but with a mixture of normal biparental cells and cells entirely of paternal isodisomic origin. We describe an infant who primarily showed signs of Beckwith-Wiedemann syndrome (BWS), but also had other severe and eventually lethal medical problems, notably refractory hypoglycemia. We performed methylation studies for BWS, but incidentally for Angelman syndrome (AS) on leukocytes and in a skin FFPE sample. We also performed chromosome microarray [CNV and single-nucleotide polymorphism (SNP) array] on leukocytes. We found that the patient had hypomethylation consistent with both BWS and AS. Remarkably, this was due to mosaic paternal UPD for chromosomes 11 and 15, respectively. The SNP microarray showed mosaic paternal UPD for all chromosomes. Patients with unusual phenotypes for a typical imprinting disorder should be studied further with assays for imprinted loci on other chromosomes. Chromosomal SNP microarrays are useful in identifying patients with multiple UPDs, sometimes of the whole genome.


Assuntos
Estudo de Associação Genômica Ampla , Mosaicismo , Síndrome de Angelman/genética , Síndrome de Beckwith-Wiedemann/genética , Variações do Número de Cópias de DNA , Metilação de DNA , Feminino , Estudos de Associação Genética , Loci Gênicos , Humanos , Lactente , Polimorfismo de Nucleotídeo Único , Dissomia Uniparental/genética
4.
Clin Genet ; 85(4): 376-80, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23586500

RESUMO

We describe a family with recurrent 11q23-qter deletion Jacobsen syndrome in two affected brothers, with unique mosaic deletion 'rescue' through development of uniparental disomy (UPD) in the mother and one of the brothers. Inheritance studies show that the deleted chromosome is of maternal origin in both boys, and microarray shows a break near the ASAM gene. Parental lymphocyte chromosomes were normal. However, the mother is homozygous in lymphocytes for all loci within the deleted region in her sons, and presumably has UPD for this region. In addition, she is mosaic for the 11q deletion seen in her sons at a level of 20-30% in skin fibroblasts. We hypothesize that one of her #11 chromosomes shows fragility, that breakage at 11q23 occurred with telomeric loss in some cells, but 'rescue' from the deletion occurred in most cells by the development of mitotic UPD. She apparently carries the 11q deletion in her germ line resulting in recurrence of the syndrome. The older son is mosaic for the 11q cell line (70-88%, remainder 46,XY), and segmental UPD11 'rescue' apparently also occurred in his cytogenetically normal cells. This is a novel phenomenon restoring disomy to an individual with a chromosomal deletion.


Assuntos
Cromossomos Humanos Par 11 , Síndrome da Deleção Distal 11q de Jacobsen/genética , Dissomia Uniparental , Deleção Cromossômica , Feminino , Humanos , Masculino , Mosaicismo , Linhagem
5.
J Equine Vet Sci ; 133: 105007, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38237706

RESUMO

A two-year-old Arabian horse presented for abnormal external genitalia and dangerous stallion-like behavior was diagnosed with disorder of sexual development (DSD), also known as intersex/hermaphroditism. Standing 1-stage surgical procedure performed under sedation, and local anesthesia to concurrently eliminate stallion-like behavior, risk of neoplastic transformation of intraabdominal gonads, and to replace ambiguous external genital with a functional, and cosmetically more acceptable anatomy. Step-1) Laparoscopic abdominal exploration and gonadectomy; Step-2) Rudimentary penis resection and perineal urethrostomy. The horse tolerated surgery well (combined surgery time 185 min) with no complications. At macroscopic examination of the gonads, they resembled hypoplastic testis-like tissues. Microscopic examination confirmed presence of seminiferous tubules, Leydig and Sertoli/granulosa cells. Cytogenetic evaluation revealed a 64,XX karyotype, SRY-negative. The stallion-like behavior subsided within days post-operatively. Long-term follow-up revealed the genitoplasty site healed without urine scalding or urethral stricture. The owner satisfaction was excellent and the horse could be used post-surgery as an athlete.


Assuntos
Transtornos do Desenvolvimento Sexual , Doenças dos Cavalos , Feminino , Masculino , Cavalos , Animais , Administração de Caso , Transtornos do Desenvolvimento Sexual/genética , Transtornos do Desenvolvimento Sexual/cirurgia , Transtornos do Desenvolvimento Sexual/veterinária , Gônadas , Cariotipagem/veterinária , Cariótipo , Doenças dos Cavalos/cirurgia
6.
ACS Chem Neurosci ; 15(6): 1169-1184, 2024 03 20.
Artigo em Inglês | MEDLINE | ID: mdl-38359277

RESUMO

Voltage-gated sodium channel (NaV) inhibitors are used to treat neurological disorders of hyperexcitability such as epilepsy. These drugs act by attenuating neuronal action potential firing to reduce excitability in the brain. However, all currently available NaV-targeting antiseizure medications nonselectively inhibit the brain channels NaV1.1, NaV1.2, and NaV1.6, which potentially limits the efficacy and therapeutic safety margins of these drugs. Here, we report on XPC-7724 and XPC-5462, which represent a new class of small molecule NaV-targeting compounds. These compounds specifically target inhibition of the NaV1.6 and NaV1.2 channels, which are abundantly expressed in excitatory pyramidal neurons. They have a > 100-fold molecular selectivity against NaV1.1 channels, which are predominantly expressed in inhibitory neurons. Sparing NaV1.1 preserves the inhibitory activity in the brain. These compounds bind to and stabilize the inactivated state of the channels thereby reducing the activity of excitatory neurons. They have higher potency, with longer residency times and slower off-rates, than the clinically used antiseizure medications carbamazepine and phenytoin. The neuronal selectivity of these compounds is demonstrated in brain slices by inhibition of firing in cortical excitatory pyramidal neurons, without impacting fast spiking inhibitory interneurons. XPC-5462 also suppresses epileptiform activity in an ex vivo brain slice seizure model, whereas XPC-7224 does not, suggesting a possible requirement of Nav1.2 inhibition in 0-Mg2+- or 4-AP-induced brain slice seizure models. The profiles of these compounds will facilitate pharmacological dissection of the physiological roles of NaV1.2 and NaV1.6 in neurons and help define the role of specific channels in disease states. This unique selectivity profile provides a new approach to potentially treat disorders of neuronal hyperexcitability by selectively downregulating excitatory circuits.


Assuntos
Epilepsia , Canais de Sódio Disparados por Voltagem , Humanos , Neurônios/metabolismo , Canais de Sódio Disparados por Voltagem/metabolismo , Epilepsia/metabolismo , Encéfalo/metabolismo , Convulsões/tratamento farmacológico , Convulsões/metabolismo , Potenciais de Ação/fisiologia
7.
Eur Spine J ; 22 Suppl 3: S450-9, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23291785

RESUMO

PURPOSE: Hemangiopericytoma (HPC) is a rare tumor of the central nervous system. Primary spinal occurrence of this tumor is extremely uncommon and cases involving the intramedullary spinal cord are even more rare. The purpose of this study was to explore the clinical features, surgical strategies, outcome and pathology in a consecutive series of patients treated at a single institution. METHODS: The authors performed a retrospective review of the clinicopathological characteristics of four patients with a pathological diagnosis of spinal HPC. RESULTS: Four cases with intradural as well as intra/extra-medullary components were identified. Gross total resection with no recurrence at the operative site was achieved in the majority of patients with a spinal HPC. One patient had significant recurrence and eventually, succumbed to the disease. CONCLUSION: Increased awareness of these tumors' capability to occur intradurally and intramedullarly can help surgeons accurately diagnose and choose an effective plan of care. Gross total resection of hemangiopericytomas is the mainstay of treatment and should be pursued if feasible. Histopathology is essential to the diagnosis.


Assuntos
Hemangiopericitoma/cirurgia , Neoplasias da Medula Espinal/cirurgia , Adulto , Feminino , Hemangiopericitoma/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Procedimentos Neurocirúrgicos , Estudos Retrospectivos , Neoplasias da Medula Espinal/patologia
8.
Biomed Pharmacother ; 165: 115173, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37453200

RESUMO

Nav1.1 is an important pharmacological target as this voltage-gated sodium channel is involved in neurological and cardiac syndromes. Channel activators are actively sought to try to compensate for haploinsufficiency in several of these pathologies. Herein we used a natural source of new peptide compounds active on ion channels and screened for drugs capable to inhibit channel inactivation as a way to compensate for decreased channel function. We discovered that JzTx-34 is highly active on Nav1.1 and subsequently performed a full structure-activity relationship investigation to identify its pharmacophore. These experiments will help interpret the mechanism of action of this and formerly identified peptides as well as the future identification of new peptides. We also reveal structural determinants that make natural ICK peptides active against Nav1.1 challenging to synthesize. Altogether, the knowledge gained by this study will help facilitate the discovery and development of new compounds active on this critical ion channel target.


Assuntos
Peptídeos , Canais de Sódio Disparados por Voltagem , Humanos , Peptídeos/farmacologia , Peptídeos/química , Relação Estrutura-Atividade
9.
Res Sq ; 2023 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-37609289

RESUMO

Background: SCN8A-related disorders are a group of variable conditions caused by pathogenic variations in SCN8A. Online Mendelian Inheritance in Man (OMIM) terms them as developmental and epileptic encephalopathy 13, benign familial infantile seizures 5 or cognitive impairment with or without cerebellar ataxia. Methods: In this study, we describe clinical and genetic results on eight individuals from six families with SCN8A pathogenic variants identified via exome sequencing. Results: Clinical findings ranged from normal development with well-controlled epilepsy to significant developmental delay with treatment-resistant epilepsy. Three novel and three reported variants were observed in SCN8A. Electrophysiological analysis in transfected cells revealed a loss-of-function variant in Patient 4. Conclusions: This work expands the clinical and genotypic spectrum of SCN8A-related disorders and provides electrophysiological results on a novel loss-of-function SCN8A variant.

10.
Elife ; 112022 03 02.
Artigo em Inglês | MEDLINE | ID: mdl-35234610

RESUMO

NBI-921352 (formerly XEN901) is a novel sodium channel inhibitor designed to specifically target NaV1.6 channels. Such a molecule provides a precision-medicine approach to target SCN8A-related epilepsy syndromes (SCN8A-RES), where gain-of-function (GoF) mutations lead to excess NaV1.6 sodium current, or other indications where NaV1.6 mediated hyper-excitability contributes to disease (Gardella and Møller, 2019; Johannesen et al., 2019; Veeramah et al., 2012). NBI-921352 is a potent inhibitor of NaV1.6 (IC500.051 µM), with exquisite selectivity over other sodium channel isoforms (selectivity ratios of 756 X for NaV1.1, 134 X for NaV1.2, 276 X for NaV1.7, and >583 Xfor NaV1.3, NaV1.4, and NaV1.5). NBI-921352is a state-dependent inhibitor, preferentially inhibiting inactivatedchannels. The state dependence leads to potent stabilization of inactivation, inhibiting NaV1.6 currents, including resurgent and persistent NaV1.6 currents, while sparing the closed/rested channels. The isoform-selective profile of NBI-921352 led to a robust inhibition of action-potential firing in glutamatergic excitatory pyramidal neurons, while sparing fast-spiking inhibitory interneurons, where NaV1.1 predominates. Oral administration of NBI-921352 prevented electrically induced seizures in a Scn8a GoF mouse,as well as in wild-type mouse and ratseizure models. NBI-921352 was effective in preventing seizures at lower brain and plasma concentrations than commonly prescribed sodium channel inhibitor anti-seizure medicines (ASMs) carbamazepine, phenytoin, and lacosamide. NBI-921352 waswell tolerated at higher multiples of the effective plasma and brain concentrations than those ASMs. NBI-921352 is entering phase II proof-of-concept trials for the treatment of SCN8A-developmental epileptic encephalopathy (SCN8A-DEE) and adult focal-onset seizures.


Assuntos
Epilepsia , Canal de Sódio Disparado por Voltagem NAV1.6 , Animais , Mutação com Ganho de Função , Camundongos , Mutação , Canal de Sódio Disparado por Voltagem NAV1.6/genética , Neurônios/fisiologia , Ratos , Sódio , Bloqueadores dos Canais de Sódio/farmacologia
11.
J Med Chem ; 64(6): 2953-2966, 2021 03 25.
Artigo em Inglês | MEDLINE | ID: mdl-33682420

RESUMO

Nav1.7 is an extensively investigated target for pain with a strong genetic link in humans, yet in spite of this effort, it remains challenging to identify efficacious, selective, and safe inhibitors. Here, we disclose the discovery and preclinical profile of GDC-0276 (1) and GDC-0310 (2), selective Nav1.7 inhibitors that have completed Phase 1 trials. Our initial search focused on close-in analogues to early compound 3. This resulted in the discovery of GDC-0276 (1), which possessed improved metabolic stability and an acceptable overall pharmacokinetics profile. To further derisk the predicted human pharmacokinetics and enable QD dosing, additional optimization of the scaffold was conducted, resulting in the discovery of a novel series of N-benzyl piperidine Nav1.7 inhibitors. Improvement of the metabolic stability by blocking the labile benzylic position led to the discovery of GDC-0310 (2), which possesses improved Nav selectivity and pharmacokinetic profile over 1.


Assuntos
Azetidinas/farmacologia , Benzamidas/farmacologia , Descoberta de Drogas , Canal de Sódio Disparado por Voltagem NAV1.7/metabolismo , Sulfonamidas/farmacologia , Bloqueadores do Canal de Sódio Disparado por Voltagem/farmacologia , Animais , Azetidinas/química , Azetidinas/farmacocinética , Benzamidas/química , Benzamidas/farmacocinética , Células Cultivadas , Células HEK293 , Humanos , Piperidinas/química , Piperidinas/farmacocinética , Piperidinas/farmacologia , Ratos Sprague-Dawley , Sulfonamidas/química , Sulfonamidas/farmacocinética , Bloqueadores do Canal de Sódio Disparado por Voltagem/química , Bloqueadores do Canal de Sódio Disparado por Voltagem/farmacocinética
12.
J Exp Med ; 160(5): 1350-9, 1984 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-6208309

RESUMO

The monoclonal antibodies Genox 3.53, S1, and R1 define polymorphic epitopes localized to DQ molecules. All three antibodies showed a strong association with the HLA-DR antigens 1, 2, and w6 when tested on a panel of 68 unrelated individuals, suggesting that they all recognized the DQw1 allospecificity. However, segregation analysis and binding studies with a panel of HLA-D/DR homozygous cells indicated that these monoclonal antibodies defined two different alloantigens. Cells homozygous for DR 1, 2, or w6 expressed the epitopes defined by all three antibodies (i.e., S1, R1, and Ge) while cells homozygous for DR4 and DRw8 expressed only the S1 and R1 epitopes. Sequential immunoprecipitation analyses in S1+/R1+/Ge+ individuals, in which the three epitopes were shown by segregation analysis to be encoded by the same chromosome, revealed two distinct DQ-like molecules. While R1 and S1 appeared to reside on the same molecule, the epitope defined by Genox 3.53 was on a different molecule. Identical results were obtained with DR1-, DR2-, or DRw6-bearing cells. Thus it appears that DQw1-bearing individuals express two cis-encoded DQ-like molecules that carry distinct alloantigenic specificities.


Assuntos
Epitopos/análise , Antígenos de Histocompatibilidade Classe II/análise , Isoantígenos/imunologia , Anticorpos Monoclonais/imunologia , Reações Antígeno-Anticorpo , Precipitação Química , Epitopos/imunologia , Código Genético , Antígenos HLA-DQ , Haploidia , Antígenos de Histocompatibilidade Classe II/imunologia , Humanos
13.
J Exp Med ; 161(2): 366-77, 1985 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-3973533

RESUMO

Monoclonal antibodies (mAb) were selected for differential binding to sections of freshly frozen biopsy material of human malignant melanomas and their precursor lesions, the melanocytic nevi. Both melanomas and normal nevi expressed human Ia-like antigens, transferrin receptor and the transferrin-related molecule p97. In contrast, only 1 nevus of 21 tested expressed both glycoprotein gp75, defined by mAb 15.75, and protein p89, defined by mAb P3.58, whereas 12 of 15 melanomas tested expressed both antigens. mAb P3.58 reacted with one additional melanoma and one nevus. The expression of these two molecules therefore appears to be correlated with the appearance of the malignant phenotype of melanocytes.


Assuntos
Antígenos de Neoplasias/análise , Antígenos de Superfície/análise , Melanoma/imunologia , Proteínas de Neoplasias/análise , Animais , Anticorpos Monoclonais , Reações Antígeno-Anticorpo , Antígenos de Neoplasias/imunologia , Antígenos de Superfície/imunologia , Diferenciação Celular , Feminino , Humanos , Melanócitos/citologia , Melanócitos/imunologia , Melanócitos/patologia , Melanoma/patologia , Antígenos Específicos de Melanoma , Camundongos , Camundongos Endogâmicos BALB C , Proteínas de Neoplasias/imunologia , Nevo/imunologia , Nevo/patologia , Distribuição Tecidual
14.
J Exp Med ; 176(4): 1083-90, 1992 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-1402654

RESUMO

The assembly of the classical, polymorphic major histocompatibility complex class I molecules in the endoplasmic reticulum requires the presence of peptide ligands and beta 2-microglobulin (beta 2m). Formation of this trimolecular complex is a prerequisite for efficient transport to the cell surface, where presented peptides are scanned by T lymphocytes. The function of the other class I molecules is in dispute. The human, nonclassical class I gene, HLA-E, was found to be ubiquitously transcribed, whereas cell surface expression was difficult to detect upon transfection. Pulse chase experiments revealed that the HLA-E heavy chain in transfectants, obtained with the murine myeloma cell line P3X63-Ag8.653 (X63), displays a significant reduction in oligosaccharide maturation and intracellular transport compared with HLA-B27 in corresponding transfectants. The accordingly low HLA-E cell surface expression could be significantly enhanced by either reducing the culture temperature or by supplementing the medium with human beta 2m, suggesting inefficient binding of endogenous peptides to HLA-E. To analyze whether HLA-E binds peptides and to identify the corresponding ligands, fractions of acid-extracted material from HLA-E/X63 transfectants were separated by reverse phase HPLC and were tested for their ability to enhance HLA-E cell surface expression. Two fractions specifically increased the HLA class I expression on the HLA-E transfectant clone.


Assuntos
Genes MHC Classe I , Antígenos HLA/biossíntese , Antígenos de Histocompatibilidade Classe I/biossíntese , Animais , Transporte Biológico , Linhagem Celular , Membrana Celular/imunologia , Citometria de Fluxo , Imunofluorescência , Expressão Gênica , Antígenos HLA/genética , Antígenos HLA/isolamento & purificação , Antígeno HLA-B27/biossíntese , Antígeno HLA-B27/genética , Antígenos de Histocompatibilidade Classe I/genética , Antígenos de Histocompatibilidade Classe I/isolamento & purificação , Humanos , Substâncias Macromoleculares , Camundongos , Mieloma Múltiplo , Ligação Proteica , Transfecção , Antígenos HLA-E
15.
J Exp Med ; 156(1): 104-11, 1982 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-6177816

RESUMO

A murine monoclonal antibody directed against a human B cell surface antigen with the characteristics of HLA-DR is described. The antigen detected is tightly linked to HLA and is correlated with the alloantigen HLA-Dw/DR3. Reactivity with a fraction of Dw/DRw6 cells is also observed. The determinant recognized by this antibody has been shown to be present on the smaller molecular weight beta subunit of the HLA-DR antigen.


Assuntos
Anticorpos Monoclonais , Epitopos , Antígenos de Histocompatibilidade Classe II/imunologia , Alótipos de Imunoglobulina/imunologia , Animais , Antígenos Heterófilos/imunologia , Fenômenos Químicos , Química , Antígeno HLA-DR3 , Humanos , Melanoma/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Peso Molecular
16.
Nanotechnology ; 21(29): 295708, 2010 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-20601769

RESUMO

Gold monolayer protected clusters (MPCs) exhibit strong quantum confinement effects and size dependent electronic, optical and chemical properties. Chemical tuning of these properties can be achieved by established synthesis methods, providing an excellent system for the study of the relationship between chemical and electronic structure. In this paper, the first electronic spectra of individual Au MPCs (Au(25)) acquired by single electron tunneling force spectroscopy on non-conducting silicon dioxide surfaces are reported. A HOMO-LUMO (highest occupied molecular orbital-lowest unoccupied molecular orbital) energy gap is observed in the Au(25) spectra. Hysteretic charging of the particles is also observed while obtaining the energy spectra. The new single electron tunneling measurement methodology is described. A model explaining the measurements supports the existence of mid-HOMO-LUMO gap defect states.

17.
PLoS One ; 15(8): e0238121, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32845893

RESUMO

Variants implicated in childhood epilepsy have been identified in all four voltage-gated sodium channels that initiate action potentials in the central nervous system. Previous research has focused on the functional effects of particular variants within the most studied of these channels (NaV1.1, NaV1.2 and NaV1.6); however, there have been few comparative studies across channels to infer the impact of mutations in patients with epilepsy. Here we compare patterns of variation in patient and public databases to test the hypothesis that regions of known functional significance within voltage-gated sodium (NaV) channels have an increased burden of deleterious variants. We assessed mutational burden in different regions of the Nav channels by (1) performing Fisher exact tests on odds ratios to infer excess variants in domains, segments, and loops of each channel in patient databases versus public "control" databases, and (2) comparing the cumulative distribution of variant sites along DNA sequences of each gene in patient and public databases (i.e., independent of protein structure). Patient variant density was concordant among channels in regions known to play a role in channel function, with statistically significant higher patient variant density in S4-S6 and DIII-DIV and an excess of public variants in SI-S3, DI-DII, DII-DIII. On the other hand, channel-specific patterns of patient burden were found in the NaV1.6 inactivation gate and NaV1.1 S5-S6 linkers, while NaV1.2 and NaV1.6 S4-S5 linkers and S5 segments shared patient variant patterns that contrasted with those in NaV1.1. These different patterns may reflect different roles played by the NaV1.6 inactivation gate in action potential propagation, and by NaV1.1 S5-S6 linkers in loss of function and haploinsufficiency. Interestingly, NaV1.2 and NaV1.6 both lack amino acid substitutions over significantly long stretches in both the patient and public databases suggesting that new mutations in these regions may cause embryonic lethality or a non-epileptic disease phenotype.


Assuntos
Epilepsia/patologia , Ativação do Canal Iônico/genética , Canal de Sódio Disparado por Voltagem NAV1.1/genética , Canal de Sódio Disparado por Voltagem NAV1.2/genética , Canal de Sódio Disparado por Voltagem NAV1.6/genética , Substituição de Aminoácidos/genética , Sequência de Bases , Encéfalo/fisiologia , Epilepsia/genética , Variação Genética/genética , Humanos , Potenciais da Membrana/genética , Mutação/genética , Técnicas de Patch-Clamp , Análise de Sequência de DNA
18.
Science ; 225(4663): 745-6, 1984 Aug 17.
Artigo em Inglês | MEDLINE | ID: mdl-6463652

RESUMO

When isolated apical membrane vesicles prepared from cultured A6 epithelia were incubated in vitro with the methyl donor S-adenosylmethionine, the control rate of amiloride-inhibitable sodium transport was doubled. The methylation inhibitors 3-deazaadenosine and S-adenosyl homocysteine returned the S-adenosyl-methionine-stimulated sodium transport to control levels. Neither these agents nor adenosine affected sodium transport into control vesicles. In vesicles incubated with S-adenosyl-[3H-methyl]methionine, both membrane phospholipids and proteins were labeled, and this labeling was inhibited by deazaadenosine. In vesicles prepared from A6 cells treated with aldosterone, sodium transport was twice the control value and S-adenosylmethionine did not cause any further stimulation of transport. In those vesicles, both lipid and protein methylation were increased. These results suggest that methylation, which increases the rate of amiloride-sensitive sodium transport is involved in the action of aldosterone at the apical membrane level in epithelia.


Assuntos
Aldosterona/fisiologia , Membrana Celular/metabolismo , Sódio/metabolismo , Amilorida/farmacologia , Anfíbios , Animais , Transporte Biológico Ativo/efeitos dos fármacos , Linhagem Celular , Membrana Celular/efeitos dos fármacos , Rim/metabolismo , Metilação , S-Adenosil-Homocisteína/farmacologia , S-Adenosilmetionina/farmacologia , Tubercidina/farmacologia
19.
Nanotechnology ; 20(5): 055701, 2009 Feb 04.
Artigo em Inglês | MEDLINE | ID: mdl-19417360

RESUMO

We report the first atomic scale imaging and spectroscopic measurements of electron trap states in completely non-conducting surfaces by dynamic tunnelling force microscopy/spectroscopy. Single electrons are dynamically shuttled to/from individual states in thick films of hafnium silicate and silicon dioxide. The new method opens up surfaces that are inaccessible to the scanning tunnelling microscope for imaging and spectroscopy on an atomic scale.


Assuntos
Háfnio/química , Microscopia de Força Atômica/métodos , Nanoestruturas/química , Nanoestruturas/ultraestrutura , Silicatos/química , Espectrofotometria Atômica/métodos , Transporte de Elétrons , Elétrons , Teste de Materiais , Semicondutores , Estresse Mecânico
20.
J Med Chem ; 62(8): 4091-4109, 2019 04 25.
Artigo em Inglês | MEDLINE | ID: mdl-30943032

RESUMO

Using structure- and ligand-based design principles, a novel series of piperidyl chromane arylsulfonamide Nav1.7 inhibitors was discovered. Early optimization focused on improvement of potency through refinement of the low energy ligand conformation and mitigation of high in vivo clearance. An in vitro hepatotoxicity hazard was identified and resolved through optimization of lipophilicity and lipophilic ligand efficiency to arrive at GNE-616 (24), a highly potent, metabolically stable, subtype selective inhibitor of Nav1.7. Compound 24 showed a robust PK/PD response in a Nav1.7-dependent mouse model, and site-directed mutagenesis was used to identify residues critical for the isoform selectivity profile of 24.


Assuntos
Canal de Sódio Disparado por Voltagem NAV1.7/química , Sulfonamidas/química , Bloqueadores do Canal de Sódio Disparado por Voltagem/química , Analgésicos/química , Analgésicos/metabolismo , Analgésicos/farmacologia , Analgésicos/uso terapêutico , Animais , Sítios de Ligação , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Dor Crônica/tratamento farmacológico , Dor Crônica/patologia , Cães , Meia-Vida , Humanos , Ligantes , Masculino , Camundongos , Simulação de Acoplamento Molecular , Mutagênese Sítio-Dirigida , Canal de Sódio Disparado por Voltagem NAV1.7/genética , Canal de Sódio Disparado por Voltagem NAV1.7/metabolismo , Isoformas de Proteínas/antagonistas & inibidores , Isoformas de Proteínas/metabolismo , Ratos , Relação Estrutura-Atividade , Sulfonamidas/metabolismo , Sulfonamidas/farmacologia , Sulfonamidas/uso terapêutico , Bloqueadores do Canal de Sódio Disparado por Voltagem/metabolismo , Bloqueadores do Canal de Sódio Disparado por Voltagem/farmacologia , Bloqueadores do Canal de Sódio Disparado por Voltagem/uso terapêutico
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