RESUMO
Cellular lipid requirements are achieved through a combination of biosynthesis and import programs. Using isotope tracer analysis, we show that type I interferon (IFN) signaling shifts the balance of these programs by decreasing synthesis and increasing import of cholesterol and long chain fatty acids. Genetically enforcing this metabolic shift in macrophages is sufficient to render mice resistant to viral challenge, demonstrating the importance of reprogramming the balance of these two metabolic pathways in vivo. Unexpectedly, mechanistic studies reveal that limiting flux through the cholesterol biosynthetic pathway spontaneously engages a type I IFN response in a STING-dependent manner. The upregulation of type I IFNs was traced to a decrease in the pool size of synthesized cholesterol and could be inhibited by replenishing cells with free cholesterol. Taken together, these studies delineate a metabolic-inflammatory circuit that links perturbations in cholesterol biosynthesis with activation of innate immunity.
Assuntos
Colesterol/metabolismo , Imunidade Inata , Interferon gama/metabolismo , Transdução de Sinais , Animais , Linhagem Celular Tumoral , Humanos , Interferon beta-1b , Proteínas de Membrana/metabolismo , Ácido Mevalônico/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Proteína de Ligação a Elemento Regulador de Esterol 1/metabolismo , Proteína de Ligação a Elemento Regulador de Esterol 2/metabolismoRESUMO
Studies have shown that disrupting the formation of the ligand-RET-GFRα complex could be an effective way of treating pain and itch. Compared to traditional high-throughput screens, DNA encoded libraries (DELs) have distinguished themselves as a powerful technology for hit identification in recent years. The present work demonstrates the use of DEL technology identifying compound 16 as the first GFRa2/GFRa3 small molecule inhibitor (0.1/0.2 µM respectively) selective over RET. This molecule represents an opportunity to advance the development of small-molecule inhibitors targeting the GFRα-RET interface for the treatment of pain and itch.
Assuntos
DNA , Receptores de Fator Neurotrófico Derivado de Linhagem de Célula Glial , Bibliotecas de Moléculas Pequenas , Bibliotecas de Moléculas Pequenas/química , Bibliotecas de Moléculas Pequenas/farmacologia , Bibliotecas de Moléculas Pequenas/síntese química , Humanos , DNA/química , DNA/metabolismo , Receptores de Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Receptores de Fator Neurotrófico Derivado de Linhagem de Célula Glial/antagonistas & inibidores , Descoberta de Drogas , Relação Estrutura-Atividade , Estrutura Molecular , Relação Dose-Resposta a DrogaRESUMO
Visceral leishmaniasis causes considerable mortality and morbidity in many parts of the world. There is an urgent need for the development of new, effective treatments for this disease. Here we describe the development of an anti-leishmanial drug-like chemical series based on a pyrazolopyrimidine scaffold. The leading compound from this series (7, DDD853651/GSK3186899) is efficacious in a mouse model of visceral leishmaniasis, has suitable physicochemical, pharmacokinetic and toxicological properties for further development, and has been declared a preclinical candidate. Detailed mode-of-action studies indicate that compounds from this series act principally by inhibiting the parasite cdc-2-related kinase 12 (CRK12), thus defining a druggable target for visceral leishmaniasis.
Assuntos
Quinases Ciclina-Dependentes/antagonistas & inibidores , Leishmania donovani/efeitos dos fármacos , Leishmania donovani/enzimologia , Leishmaniose Visceral/tratamento farmacológico , Leishmaniose Visceral/parasitologia , Terapia de Alvo Molecular , Pirazóis/farmacologia , Pirimidinas/farmacologia , Animais , Quinase 9 Dependente de Ciclina/química , Quinases Ciclina-Dependentes/química , Quinases Ciclina-Dependentes/metabolismo , Modelos Animais de Doenças , Humanos , Camundongos , Simulação de Acoplamento Molecular , Proteoma/efeitos dos fármacos , Proteômica , Pirazóis/química , Pirazóis/uso terapêutico , Pirimidinas/química , Pirimidinas/uso terapêutico , Reprodutibilidade dos Testes , Especificidade por SubstratoRESUMO
This article explores the impact of combining tissue viability and lymphoedema techniques on optimising time to healing. AIM: To investigate the healing rates observed in patients who presented to wound and lymphoedema specialist clinics, located in the south eastern region of England, with venous/lymphovenous ulceration of the lower limb during the COVID-19 pandemic in 2020-2022 (30 months in all). METHODOLOGY: A retrospective analysis of patient outcomes. RESULTS: 1041 patients were referred to the service, with a healing rate of 88.5% over 78 days. DISCUSSION: When comparing 2013-2019 healing rates/time to healing vs 2020-2022 there was a decrease of 1.5% in the rate of healing and a mean reduction in time to healing of 40 days. CONCLUSION: Despite the pandemic the service was able to maintain previous levels of outcomes and observed a decrease in the mean time to healing.
Assuntos
COVID-19 , Linfedema , Úlcera Varicosa , Humanos , Pandemias , Estudos Retrospectivos , Inglaterra/epidemiologia , Extremidade InferiorRESUMO
Developmental delay and intellectual disability (DD and ID) are heterogeneous phenotypes that arise in many rare monogenic disorders. Because of this rarity, developing cohorts with enough individuals to robustly identify disease-associated genes is challenging. Social-media platforms that facilitate data sharing among sequencing labs can help to address this challenge. Through one such tool, GeneMatcher, we identified nine DD- and/or ID-affected probands with a rare, heterozygous variant in the gene encoding the serine/threonine-protein kinase BRSK2. All probands have a speech delay, and most present with intellectual disability, motor delay, behavioral issues, and autism. Six of the nine variants are predicted to result in loss of function, and computational modeling predicts that the remaining three missense variants are damaging to BRSK2 structure and function. All nine variants are absent from large variant databases, and BRSK2 is, in general, relatively intolerant to protein-altering variation among humans. In all six probands for whom parents were available, the mutations were found to have arisen de novo. Five of these de novo variants were from cohorts with at least 400 sequenced probands; collectively, the cohorts span 3,429 probands, and the observed rate of de novo variation in these cohorts is significantly higher than the estimated background-mutation rate (p = 2.46 × 10-6). We also find that exome sequencing provides lower coverage and appears less sensitive to rare variation in BRSK2 than does genome sequencing; this fact most likely reduces BRSK2's visibility in many clinical and research sequencing efforts. Altogether, our results implicate damaging variation in BRSK2 as a source of neurodevelopmental disease.
Assuntos
Deficiências do Desenvolvimento/genética , Deleção de Genes , Deficiência Intelectual/genética , Transtornos do Neurodesenvolvimento/genética , Proteínas Serina-Treonina Quinases/genética , Adolescente , Transtorno Autístico/genética , Criança , Transtornos do Comportamento Infantil/genética , Pré-Escolar , Exoma , Feminino , Predisposição Genética para Doença , Variação Genética , Heterozigoto , Humanos , Masculino , Transtornos das Habilidades Motoras/genética , Mutação , Fenótipo , Sequenciamento do Exoma , Adulto JovemRESUMO
Following the modern synthesis, mating signals were thought of principally as species recognition traits, a view later challenged by a burgeoning interest in sexual selection-specifically mate choice. In the 1990s, these different signal functions were proposed to represent a single process driven by the shape of female preference functions across both intra- and interspecific signal space. However, the properties of reliable 'recognition' signals (stereotyped; low intraspecific variation) and informative 'quality' signals (condition dependent; high intraspecific variation) seem at odds, perhaps favouring different signal components for different functions. Surprisingly, the idea that different components of mating signals are evaluated in series, first to recognize generally compatible mates and then to select for quality, has never been explicitly tested. Here I evaluate patterns of (i) intraspecific signal variation, (ii) female preference function shape and (iii) phylogenetic signal for male cricket call components known to be processed in series. The results show that signal components processed first tend to have low variation, closed preference functions and low phylogenetic signal, whereas signal components processed later show the opposite, suggesting that mating signal evaluation follows an 'order-of-operations'. Applicability of this finding to diverse groups of organisms and sensory modalities is discussed.
Assuntos
Preferência de Acasalamento Animal , Animais , Feminino , Masculino , Fenótipo , Filogenia , Reprodução , Comportamento Sexual Animal , Seleção SexualRESUMO
PURPOSE: SouthSeq is a translational research study that undertook genome sequencing (GS) for infants with symptoms suggestive of a genetic disorder. Recruitment targeted racial/ethnic minorities and rural, medically underserved areas in the Southeastern United States, which are historically underrepresented in genomic medicine research. METHODS: GS and analysis were performed for 367 infants to detect disease-causal variation concurrent with standard of care evaluation and testing. RESULTS: Definitive diagnostic (DD) or likely diagnostic (LD) genetic findings were identified in 30% of infants, and 14% of infants harbored an uncertain result. Only 43% of DD/LD findings were identified via concurrent clinical genetic testing, suggesting that GS testing is better for obtaining early genetic diagnosis. We also identified phenotypes that correlate with the likelihood of receiving a DD/LD finding, such as craniofacial, ophthalmologic, auditory, skin, and hair abnormalities. We did not observe any differences in diagnostic rates between racial/ethnic groups. CONCLUSION: We describe one of the largest-to-date GS cohorts of ill infants, enriched for African American and rural patients. Our results show the utility of GS because it provides early-in-life detection of clinically relevant genetic variations not detected by current clinical genetic testing, particularly for infants exhibiting certain phenotypic features.
Assuntos
Testes Diagnósticos de Rotina , Testes Genéticos , Sequência de Bases , Mapeamento Cromossômico , Testes Genéticos/métodos , Genômica , HumanosRESUMO
Visceral leishmaniasis (VL), caused by the protozoan parasites Leishmania donovani and Leishmania infantum, is one of the major parasitic diseases worldwide. There is an urgent need for new drugs to treat VL, because current therapies are unfit for purpose in a resource-poor setting. Here, we describe the development of a preclinical drug candidate, GSK3494245/DDD01305143/compound 8, with potential to treat this neglected tropical disease. The compound series was discovered by repurposing hits from a screen against the related parasite Trypanosoma cruzi Subsequent optimization of the chemical series resulted in the development of a potent cidal compound with activity against a range of clinically relevant L. donovani and L. infantum isolates. Compound 8 demonstrates promising pharmacokinetic properties and impressive in vivo efficacy in our mouse model of infection comparable with those of the current oral antileishmanial miltefosine. Detailed mode of action studies confirm that this compound acts principally by inhibition of the chymotrypsin-like activity catalyzed by the ß5 subunit of the L. donovani proteasome. High-resolution cryo-EM structures of apo and compound 8-bound Leishmania tarentolae 20S proteasome reveal a previously undiscovered inhibitor site that lies between the ß4 and ß5 proteasome subunits. This induced pocket exploits ß4 residues that are divergent between humans and kinetoplastid parasites and is consistent with all of our experimental and mutagenesis data. As a result of these comprehensive studies and due to a favorable developability and safety profile, compound 8 is being advanced toward human clinical trials.
Assuntos
Antiprotozoários/administração & dosagem , Leishmania donovani/efeitos dos fármacos , Leishmania infantum/efeitos dos fármacos , Leishmaniose Visceral/diagnóstico por imagem , Inibidores de Proteassoma/administração & dosagem , Proteínas de Protozoários/antagonistas & inibidores , Animais , Antiprotozoários/química , Sítios de Ligação , Modelos Animais de Doenças , Avaliação Pré-Clínica de Medicamentos , Humanos , Leishmania donovani/química , Leishmania donovani/enzimologia , Leishmania infantum/química , Leishmania infantum/enzimologia , Leishmaniose Visceral/parasitologia , Masculino , Camundongos , Complexo de Endopeptidases do Proteassoma/química , Complexo de Endopeptidases do Proteassoma/metabolismo , Inibidores de Proteassoma/química , Conformação Proteica , Proteínas de Protozoários/química , Proteínas de Protozoários/metabolismoRESUMO
The paper engages Christopher Boorse's Bio-Statistical Theory (BST). In its current form, BST runs into a significant challenge. For BST to account for its central tenet-that lower-level part-dysfunction is sufficient for higher-level pathology-it must provide criteria for how to decide which lower-level parts are the ones to be analyzed for health or pathology. As BST is a naturalistic theory, such choices must be based solely on naturalistic considerations. An argument is provided to show that, if BST is to be preserved, such parthood choices are based on non-naturalistic considerations. We demonstrate that even when parthood choices are based on the best way to preserve BST, there are counterintuitive results which bring the central tenet of BST into question.
Assuntos
Doença , Filosofia Médica , Dissidências e Disputas , Saúde , HumanosRESUMO
PURPOSE: To evaluate the effectiveness and specificity of population-based genomic screening in Alabama. METHODS: The Alabama Genomic Health Initiative (AGHI) has enrolled and evaluated 5369 participants for the presence of pathogenic/likely pathogenic (P/LP) variants using the Illumina Global Screening Array (GSA), with validation of all P/LP variants via Sanger sequencing in a CLIA-certified laboratory before return of results. RESULTS: Among 131 variants identified by the GSA that were evaluated by Sanger sequencing, 67 (51%) were false positives (FP). For 39 of the 67 FP variants, a benign/likely benign variant was present at or near the targeted P/LP variant. Variants detected within African American individuals were significantly enriched for FPs, likely due to a higher rate of nontargeted alternative alleles close to array-targeted P/LP variants. CONCLUSION: In AGHI, we have implemented an array-based process to screen for highly penetrant genetic variants in actionable disease genes. We demonstrate the need for clinical validation of array-identified variants in direct-to-consumer or population testing, especially for diverse populations.
Assuntos
Testes Genéticos , Genômica , Alabama , Variação Genética , Sequenciamento de Nucleotídeos em Larga Escala , HumanosRESUMO
BACKGROUND: Dopamine D1 receptor signaling plays key roles in core domains of neural function, including cognition and reward processing; however, many questions remain about the functions of circuits modulated by dopamine D1 receptor, largely because clinically viable, selective agonists have yet to be tested in humans. METHODS: Using a novel, exploratory neurofunctional domains study design, we assessed the safety, tolerability, pharmacodynamics, and pharmacokinetics of PF-06412562, a selective D1/D5R partial agonist, in healthy male volunteers who met prespecified criteria for low working memory capacity. Functional magnetic resonance imaging, electrophysiologic endpoints, and behavioral paradigms were used to assess working memory, executive function, and motivation/reward processing following multiple-dose administration of PF-06412562. A total of 77 patients were assigned PF-06412562 (3 mg twice daily and 15 mg twice daily) or placebo administered for 5 to 7 days. Due to the exploratory nature of the study, it was neither powered for any specific treatment effect nor corrected for multiple comparisons. RESULTS: Nominally significant improvements from baseline in cognitive endpoints were observed in all 3 groups; however, improvements in PF-06412562-treated patients were less than in placebo-treated participants. Motivation/reward processing endpoints were variable. PF-06412562 was safe and well tolerated, with no serious adverse events, severe adverse events, or adverse events leading to dose reduction or temporary discontinuation except for 1 permanent discontinuation due to increased orthostatic heart rate. CONCLUSIONS: PF-06412562, in the dose range and patient population explored in this study, did not improve cognitive function or motivation/reward processing more than placebo over the 5- to 7-day treatment period. CLINICALTRIALS.GOV IDENTIFIER: NCT02306876.
Assuntos
Encéfalo/efeitos dos fármacos , Cognição/efeitos dos fármacos , Agonistas de Dopamina/administração & dosagem , Memória de Curto Prazo/efeitos dos fármacos , Motivação/efeitos dos fármacos , Receptores de Dopamina D1/agonistas , Receptores de Dopamina D5/agonistas , Adolescente , Adulto , Encéfalo/diagnóstico por imagem , Encéfalo/metabolismo , Agonistas de Dopamina/efeitos adversos , Agonistas de Dopamina/farmacocinética , Método Duplo-Cego , Esquema de Medicação , Agonismo Parcial de Drogas , Função Executiva/efeitos dos fármacos , Voluntários Saudáveis , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Receptores de Dopamina D1/metabolismo , Receptores de Dopamina D5/metabolismo , Adulto JovemRESUMO
COTI (collar therapy indicator) has been recently introduced for the detection of gamma rays with emphasis on thyroid investigations. The aim of this study was to test the feasibility of a prototype version of COTI including activity detectors with low sensitivity in performing thyroid uptake measurements for a large group of patients. Consequently, thyroid uptake tests were carried out for a total of 89 patients (22 males and 67 females; age: 44 ± 13 years) with thyroid cancer (n = 74), hyperthyroidism (n = 16) at 2 and 24 h after administration of 0.44-2 MBq of 131I. Eight individuals among the thyroid cancer patients were monitored up to 96 h after administration. The COTI device was equipped with two CsI (Tl) detectors, known as LoHi type, sensitive to activity ranges from 0.02 to 30 MBq of 131I. The uptake values from COTI were compared with those measured with a standard probe. It was found that the mean uptake of thyroid activity in thyroid cancer patients was 2.1 ± 1.3% at 2 h when measured with the standard probe, while it was 2.2 ± 1.2% when measured with COTI. In addition, the average uptake at 24 h after administration was 2.5 ± 3.2% and 3.2 ± 3.8% measured with COTI and the standard probe, respectively. A strong correlation was found at 24 h between the results obtained with COTI and the standard probe, while a weaker correlation was seen at 2 h. Overall, there was no significant difference between the results obtained with the standard probe and those obtained with COTI at both 2 and 24 h (Pvalue ≥ 0.05). Besides, 85% of the uptake values measured with COTI were less than those measured with the standard probe at the 24 h after administration. The average uptake value was 0.9 ± 0.8% after 96 h by COTI, and 1.4 ± 1.3% by the standard probe. Pertaining to the hyperthyroidism patients, COTI showed mean uptake values of 20 ± 16% and 23 ± 18% at 2 and 24 h, respectively. In contrast, the standard probe suggested higher mean uptake values of 26 ± 18% and 30 ± 22%, respectively. It is concluded that the prototype of COTI used in the present study has been proved to be a feasible and promising tool in thyroid investigations. It is noted, however, that the next COTI generation should include detectors equipped with collimator and energy discrimination.
Assuntos
Radioisótopos do Iodo , Traçadores Radioativos , Glândula Tireoide/metabolismo , Administração Oral , Adulto , Feminino , Humanos , Hipertireoidismo/metabolismo , Hipertireoidismo/radioterapia , Hipertireoidismo/cirurgia , Masculino , Pessoa de Meia-Idade , Dosagem Radioterapêutica , Neoplasias da Glândula Tireoide/metabolismo , Neoplasias da Glândula Tireoide/radioterapia , Neoplasias da Glândula Tireoide/cirurgia , TireoidectomiaRESUMO
Chagas' disease, which is caused by the Trypanosoma cruzi parasite, has become a global health problem that is currently treated with poorly tolerated drugs that require prolonged dosing. Therefore, there is a clinical need for new therapeutic agents that can mitigate these issues. The phosphomannomutase (PMM) and GDP-mannose pyrophosphorylase (GDP-MP) enzymes form part of the de novo biosynthetic pathway to the nucleotide sugar GDP-mannose. This nucleotide sugar is used either directly, or indirectly via the formation of dolichol-phosphomannose, for the assembly of all mannose-containing glycoconjugates. In T. cruzi, mannose-containing glycoconjugates include the cell-surface glycoinositol-phospholipids and the glycosylphosphatidylinositol-anchored mucin-like glycoproteins that dominate the cell surface architectures of all life cycle stages. This makes PMM and GDP-MP potentially attractive targets for a drug discovery program against Chagas' disease. To assess the ligandability of these enzymes in T. cruzi, we have screened 18,117 structurally diverse compounds exploring drug-like chemical space and 16,845 small polar fragment compounds using an assay interrogating the activities of both PMM and GDP-MP enzymes simultaneously. This resulted in 48 small fragment hits, and on retesting 20 were found to be active against the enzymes. Deconvolution revealed that these were all inhibitors of T. cruzi GDP-MP, with compounds 2 and 3 acting as uncompetitive and competitive inhibitors, respectively. Based on these findings, the T. cruzi PMM and GDP-MP enzymes were deemed not ligandable and poorly ligandable, respectively, using small molecules from conventional drug discovery chemical space. This presents a significant hurdle to exploiting these enzymes as therapeutic targets for Chagas' disease.
Assuntos
Antiprotozoários/farmacologia , Manose/metabolismo , Nucleotidiltransferases/metabolismo , Fosfotransferases (Fosfomutases)/metabolismo , Trypanosoma cruzi/enzimologia , Doença de Chagas/parasitologia , Descoberta de Drogas/métodos , Manosefosfatos/metabolismo , Nucleotidiltransferases/genética , Fosfotransferases (Fosfomutases)/genéticaRESUMO
Chagas' disease, caused by the protozoan parasite Trypanosoma cruzi, is a potentially life-threatening condition that has become a global issue. Current treatment is limited to two medicines that require prolonged dosing and are associated with multiple side effects, which often lead to treatment discontinuation and failure. One way to address these shortcomings is through target-based drug discovery on validated T. cruzi protein targets. One such target is the proteasome, which plays a crucial role in protein degradation and turnover through chymotrypsin-, trypsin-, and caspase-like catalytic activities. In order to initiate a proteasome drug discovery program, we isolated proteasomes from T. cruzi epimastigotes and characterized their activity using a commercially available glow-like luminescence-based assay. We developed a high-throughput biochemical assay for the chymotrypsin-like activity of the T. cruzi proteasome, which was found to be sensitive, specific, and robust but prone to luminescence technology interference. To mitigate this, we also developed a counterscreen assay that identifies potential interferers at the levels of both the luciferase enzyme reporter and the mechanism responsible for a glow-like response. Interestingly, we also found that the peptide substrate for chymotrypsin-like proteasome activity was not specific and was likely partially turned over by other catalytic sites of the protein. Finally, we utilized these biochemical tools to screen 18,098 compounds, exploring diverse drug-like chemical space, which allowed us to identify 39 hits that were active in the primary screening assay and inactive in the counterscreen assay.
Assuntos
Ensaios de Triagem em Larga Escala/métodos , Complexo de Endopeptidases do Proteassoma/metabolismo , Inibidores de Proteassoma/farmacologia , Tripanossomicidas/farmacologia , Trypanosoma cruzi/efeitos dos fármacos , Sistema Livre de Células , Luminescência , Proteínas de Protozoários/antagonistas & inibidores , Proteínas de Protozoários/metabolismo , Reprodutibilidade dos Testes , Trypanosoma cruzi/químicaRESUMO
Leishmaniasis is a neglected disease that is caused by different species of the protozoan parasite Leishmania, and it currently affects 12 million people worldwide. The antileishmanial therapeutic arsenal remains very limited in number and efficacy, and there is no vaccine for this parasitic disease. One pathway that has been genetically validated as an antileishmanial drug target is the biosynthesis of uridine diphosphate-glucose (UDP-Glc), and its direct derivative UDP-galactose (UDP-Gal). De novo biosynthesis of these two nucleotide sugars is controlled by the specific UDP-glucose pyrophosphorylase (UGP). Leishmania parasites additionally express a UDP-sugar pyrophosphorylase (USP) responsible for monosaccharides salvage that is able to generate both UDP-Gal and UDP-Glc. The inactivation of the two parasite pyrophosphorylases UGP and USP, results in parasite death. The present study reports on the identification of structurally diverse scaffolds for the development of USP inhibitors by fragment library screening. Based on this screening, we selected a small set of commercially available compounds, and identified molecules that inhibit both Leishmania major USP and UGP, with a half-maximal inhibitory concentration in the 100 µM range. The inhibitors were predicted to bind at allosteric regulation sites, which were validated by mutagenesis studies. This study sets the stage for the development of potent USP inhibitors.
Assuntos
Leishmania major/efeitos dos fármacos , Bibliotecas de Moléculas Pequenas/química , UTP-Glucose-1-Fosfato Uridililtransferase/antagonistas & inibidores , Técnicas Biossensoriais , Descoberta de Drogas , Avaliação Pré-Clínica de Medicamentos , Humanos , Cinética , Simulação de Acoplamento Molecular , Açúcares de Uridina DifosfatoRESUMO
PURPOSE: Clinically relevant secondary variants were identified in parents enrolled with a child with developmental delay and intellectual disability. METHODS: Exome/genome sequencing and analysis of 789 "unaffected" parents was performed. RESULTS: Pathogenic/likely pathogenic variants were identified in 21 genes within 25 individuals (3.2%), with 11 (1.4%) participants harboring variation in a gene defined as clinically actionable by the American College of Medical Genetics and Genomics. These 25 individuals self-reported either relevant clinical diagnoses (5); relevant family history or symptoms (13); or no relevant family history, symptoms, or clinical diagnoses (7). A limited carrier screen was performed yielding 15 variants in 48 (6.1%) parents. Parents were also analyzed as mate pairs (n = 365) to identify cases in which both parents were carriers for the same recessive disease, yielding three such cases (0.8%), two of which had children with the relevant recessive disease. Four participants had two findings (one carrier and one noncarrier variant). In total, 71 of the 789 enrolled parents (9.0%) received secondary findings. CONCLUSION: We provide an overview of the rates and types of clinically relevant secondary findings, which may be useful in the design and implementation of research and clinical sequencing efforts to identify such findings.
Assuntos
Sequenciamento do Exoma , Exoma/genética , Doenças Genéticas Inatas/genética , Testes Genéticos , Adulto , Mapeamento Cromossômico , Feminino , Triagem de Portadores Genéticos , Doenças Genéticas Inatas/classificação , Doenças Genéticas Inatas/fisiopatologia , Variação Genética , Genoma Humano/genética , Humanos , Masculino , Pessoa de Meia-Idade , Mutação , Pais , Sequenciamento Completo do GenomaRESUMO
PURPOSE: As massively parallel sequencing is increasingly being used for clinical decision making, it has become critical to understand parameters that affect sequencing quality and to establish methods for measuring and reporting clinical sequencing standards. In this report, we propose a definition for reduced coverage regions and describe a set of standards for variant calling in clinical sequencing applications. METHODS: To enable sequencing centers to assess the regions of poor sequencing quality in their own data, we optimized and used a tool (ExCID) to identify reduced coverage loci within genes or regions of particular interest. We used this framework to examine sequencing data from 500 patients generated in 10 projects at sequencing centers in the National Human Genome Research Institute/National Cancer Institute Clinical Sequencing Exploratory Research Consortium. RESULTS: This approach identified reduced coverage regions in clinically relevant genes, including known clinically relevant loci that were uniquely missed at individual centers, in multiple centers, and in all centers. CONCLUSION: This report provides a process road map for clinical sequencing centers looking to perform similar analyses on their data.
Assuntos
Sequenciamento do Exoma/métodos , Análise de Sequência de DNA/métodos , Sequenciamento Completo do Genoma/métodos , Sequência de Bases , Mapeamento Cromossômico , Exoma , Genoma Humano , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Humanos , Análise de Sequência de DNA/normas , SoftwareRESUMO
Vestigial morphological traits are common and well known in a variety of taxa. Identification of vestigial genes has illustrated the potential for evolutionary reversals and the re-expression of atavistic traits. Here we induce expression of a behavioural sexual signal, male calling song, in a cricket species, Gryllus ovisopis, which lacks a functional calling song. We successfully used acetylcholine injections in the frontal space of the head of male crickets to activate cerebral command neurons for cricket calling, and we recorded calling songs with a temporal chirp pattern similar to that of G. ovisopis' close evolutionary relatives, G. firmus and G. pennsylvanicus, implying that the neural pattern generators that underlie cricket calling behaviour persist in a vestigial state in G. ovisopis To our knowledge, this is the first demonstration of the induced expression of a vestigial behaviour in any organism. The retention of latent neural capacity to express sexual behaviours could have important implications for rapid evolution, trait re-emergence and reproductive isolation.
Assuntos
Gryllidae/fisiologia , Vocalização Animal/fisiologia , Acetilcolina/administração & dosagem , Acetilcolina/farmacologia , Animais , Gryllidae/efeitos dos fármacos , Masculino , Comportamento Sexual/efeitos dos fármacos , Vocalização Animal/efeitos dos fármacosRESUMO
A three-step synthetic route to a structurally unique π-expanded pyrrolo[3,2-b]pyrrole derived bis-ketone has been developed. In contrast to all previous ladder-type pyrrolopyrroles, the new dye exhibits a low-energy absorption band in the visible region which is responsible for its red-purple color. Interestingly, even though the compound is centrosymmetric, this band coincides with the lowest energy two-photon absorption (TPA) transition. This non-typical behaviour has been computationally rationalized by finding two close lying excited states, one of which (S1) is active for OPA and the other (S2) for TPA processes, which arise from the mixing of two symmetric partial charge-transfer states. The ultrafast excited-state dynamics was characterized by means of transient absorption analysis. A relaxation process involving S1 symmetry breaking occurs in a few ps, leading to the formation of the lowest energy charge-transfer state. This is weakly emitting, with a measured lifetime in the order of tens of picoseconds. Interestingly, two-photon polymerization has been achieved using this new ketone. The high yield of radical photo-initiation upon two-photon excitation was demonstrated by the fabrication of woodpile photonic crystal templates by direct laser writing using a zirconium-silicon hybrid composite.