RESUMO
Clustered regularly interspaced short palindromic repeats (CRISPR) technology has revolutionized creating targeted genetic variation in crops. Although CRISPR enzymes have been reported to have high sequence-specificity, careful design of the editing reagents can also reduce unintended edits at highly homologous sites. This work details the first large-scale study of the heritability of on-target edits and the rate of edits at off-target sites in soybean (Glycine max), assaying ~700 T1 plants each resulting from transformation with LbCas12a constructs containing CRISPR RNAs (crRNAs) predicted to be either "unique" with no off-target sites or "promiscuous" with >10 potential off-targets in the soybean genome. Around 80% of the on-target edits observed in T0 plants were inherited in the T1 generation, and ~49% of the total observed on-target edits in T1 were not observed at T0, indicating continued activity of LbCas12a throughout the life cycle of the plant. In planta editing at off-target sites was observed for the Promiscuous but not the Unique crRNA. Examination of the edited off-target sites revealed that LbCas12a was highly tolerant to mismatches between the crRNA and target site in bases 21-23 relative to the start of the protospacer, but even a single mismatch in the first 20 nt drastically reduced the editing rate. In addition, edits at off-target sites have lower inheritance rates than on-target edits, suggesting that they occur later in the plant's lifecycle. Plants with a desired on-target edit and no off-target edits could be identified in the T1 generation for 100% of the T0 plants edited with the Unique crRNA compared with the 65% of T0 plants edited with the Promiscuous crRNA. This confirms that proper crRNA selection can reduce or eliminate off-target editing. Even when potential off-target sites are predicted, plants containing only the intended edits can still be identified and propagated.
RESUMO
Hybrid strategies that combine conventional top-down lithography with bottom-up molecular assembly are of interest for a range of applications including nanolithography and sensors. Interest in these strategies stems from the ability to create complex architectures over large areas with molecular-scale control and precision. The molecular-ruler process typifies this approach where the sequential layer-by-layer assembly of mercaptoalkanoic acid molecules and metal ions are combined with conventional top-down lithography to create precise, registered nanogaps. However, the quality of the metal-ligated mercaptoalkanoic acid multilayer is a critical characteristic in generating reproducible and robust nanoscale structures via the molecular-ruler process. Therefore, we explore the assembly of alkanethiolate monolayers, mercaptohexadecanoic acid (MHDA) monolayers, and Cu-ligated MHDA multilayers on Au{111} substrates using atomic force microscopy and in situ dynamic spectroscopic ellipsometry. The chemical film thicknesses in situ dynamic spectroscopic ellipsometry agree with previous ex situ surface analytical methods. Moreover, in situ dynamic spectroscopic ellipsometry provides insight into the assembly process without interrupting the assembly process and potentially altering the characteristics of the resulting chemical film. By following the real-time dynamics of each deposition step, the assembly of the Cu-ligated MHDA multilayers can be optimized to minimize deposition time while having minimal impact to the quality of the chemical film.
RESUMO
Recognition of distinct phenotypic features is an important component of genetic diagnosis. Although CHARGE syndrome, Kabuki syndrome, and a recently delineated KMT2D Ex 38/39 allelic disorder exhibit significant overlap, differences on neuroimaging may help distinguish these conditions and guide genetic testing and variant interpretation. We present an infant clinically diagnosed with CHARGE syndrome but subsequently found to have a de novo missense variant in exon 38 of KMT2D, the gene implicated in both Kabuki syndrome and a distinct KMT2D allelic disorder. We compare her brain and inner ear morphology to a retrospective cohort of 21 patients with classic Kabuki syndrome and to typical CHARGE syndrome findings described in the literature. Thirteen of the 21 Kabuki syndrome patients had temporal bone imaging (5/13 CT, 12/13 MRI) and/or brain MRI (12/13) which revealed findings distinct from both CHARGE syndrome and the KMT2D allelic disorder. Our findings further elucidate the spectrum of inner ear dysmorphology distinguishing Kabuki syndrome and the KMT2D allelic disorder from CHARGE syndrome, suggesting that these three disorders may be differentiated at least in part by their inner ear anomalies.
Assuntos
Anormalidades Múltiplas/genética , Síndrome CHARGE/genética , Proteínas de Ligação a DNA/genética , Face/anormalidades , Predisposição Genética para Doença , Doenças Hematológicas/genética , Proteínas de Neoplasias/genética , Doenças Vestibulares/genética , Anormalidades Múltiplas/diagnóstico por imagem , Anormalidades Múltiplas/patologia , Síndrome CHARGE/diagnóstico por imagem , Síndrome CHARGE/patologia , DNA Helicases/genética , Face/diagnóstico por imagem , Face/patologia , Feminino , Doenças Hematológicas/diagnóstico por imagem , Doenças Hematológicas/patologia , Histona Desmetilases/genética , Humanos , Lactente , Recém-Nascido , Masculino , Mutação/genética , Neuroimagem , Fenótipo , Estudos Retrospectivos , Doenças Vestibulares/diagnóstico por imagem , Doenças Vestibulares/patologiaRESUMO
PURPOSE: Contouring variation is one of the largest systematic uncertainties in radiotherapy, yet its effect on clinical outcome has never been analyzed quantitatively. We propose a novel, robust methodology to locally quantify target contour variation in a large patient cohort and find where this variation correlates with treatment outcome. We demonstrate its use on biochemical recurrence for prostate cancer patients. METHOD: We propose to compare each patient's target contours to a consistent and unbiased reference. This reference was created by auto-contouring each patient's target using an externally trained deep learning algorithm. Local contour deviation measured from the reference to the manual contour was projected to a common frame of reference, creating contour deviation maps for each patient. By stacking the contour deviation maps, time to event was modeled pixel-wise using a multivariate Cox proportional hazards model (CPHM). Hazard ratio (HR) maps for each covariate were created, and regions of significance found using cluster-based permutation testing on the z-statistics. This methodology was applied to clinical target volume (CTV) contours, containing only the prostate gland, from 232 intermediate- and high-risk prostate cancer patients. The reference contours were created using ADMIRE® v3.4 (Elekta AB, Sweden). Local contour deviations were computed in a spherical coordinate frame, where differences between reference and clinical contours were projected in a 2D map corresponding to sampling across the coronal and transverse angles every 3°. Time to biochemical recurrence was modeled using the pixel-wise CPHM analysis accounting for contour deviation, patient age, Gleason score, and treated CTV volume. RESULTS: We successfully applied the proposed methodology to a large patient cohort containing data from 232 patients. In this patient cohort, our analysis highlighted regions where the contour variation was related to biochemical recurrence, producing expected and unexpected results: (a) the interface between prostate-bladder and prostate-seminal vesicle interfaces where increase in the manual contour relative to the reference was related to a reduction of risk of biochemical recurrence by 4-8% per mm and (b) the prostate's right, anterior and posterior regions where an increase in the manual contour relative to the reference contours was related to an increase in risk of biochemical recurrence by 8-24% per mm. CONCLUSION: We proposed and successfully applied a novel methodology to explore the correlation between contour variation and treatment outcome. We analyzed the effect of contour deviation of the prostate CTV on biochemical recurrence for a cohort of more than 200 prostate cancer patients while taking basic clinical variables into account. Applying this methodology to a larger dataset including additional clinically important covariates and externally validating it can more robustly identify regions where contour variation directly relates to treatment outcome. For example, in the prostate case we use to demonstrate our novel methodology, external validation will help confirm or reject the counter-intuitive results (larger contours resulting in higher risk). Ultimately, the results of this methodology could inform contouring protocols based on actual patient outcomes.
Assuntos
Neoplasias da Próstata , Planejamento da Radioterapia Assistida por Computador , Humanos , Masculino , Neoplasias da Próstata/diagnóstico por imagem , Neoplasias da Próstata/radioterapia , Suécia , Resultado do TratamentoRESUMO
Importance: A targeted genomic sequencing platform focused on diseases presenting in the first year of life may minimize financial and ethical challenges associated with rapid whole-genomic sequencing. Objective: To report interim variants and associated interpretations of an ongoing study comparing rapid whole-genomic sequencing with a novel targeted genomic platform composed of 1722 actionable genes targeting disorders presenting in infancy. Design, Setting, and Participants: The Genomic Medicine in Ill Neonates and Infants (GEMINI) study is a prospective, multicenter clinical trial with projected enrollment of 400 patients. The study is being conducted at 6 US hospitals. Hospitalized infants younger than 1 year of age suspected of having a genetic disorder are eligible. Results of the first 113 patients enrolled are reported here. Patient recruitment began in July 2019, and the interim analysis of enrolled patients occurred from March to June 2020. Interventions: Patient (proband) and parents (trios, when available) were tested simultaneously on both genomic platforms. Each laboratory performed its own phenotypically driven interpretation and was blinded to other results. Main Outcomes and Measures: Variants were classified according to the American College of Medical Genetics and Genomics standards of pathogenic (P), likely pathogenic (LP), or variants of unknown significance (VUS). Chromosomal and structural variations were reported by rapid whole-genomic sequencing. Results: Gestational age of 113 patients ranged from 23 to 40 weeks and postmenstrual age from 27 to 83 weeks. Sixty-seven patients (59%) were male. Diagnostic and/or VUS were returned for 51 patients (45%), while 62 (55%) had negative results. Results were concordant between platforms in 83 patients (73%). Thirty-seven patients (33%) were found to have a P/LP variant by 2 or both platforms and 14 (12%) had a VUS possibly related to phenotype. The median day of life at diagnosis was 22 days (range, 3-313 days). Significant alterations in clinical care occurred in 29 infants (78%) with a P/LP variant. Incidental findings were reported in 7 trios. Of 51 positive cases, 34 (67%) differed in the reported result because of technical limitations of the targeted platform, interpretation of the variant, filtering discrepancies, or multiple causes. Conclusions and Relevance: As comprehensive genetic testing becomes more routine, these data highlight the critically important variant detection capabilities of existing genomic sequencing technologies and the significant limitations that must be better understood.
Assuntos
Doenças Genéticas Inatas/diagnóstico , Testes Genéticos/métodos , Variação Genética , Sequenciamento Completo do Genoma , Feminino , Medicina Genômica , Humanos , Lactente , Recém-Nascido , Masculino , Estudos Prospectivos , Estados UnidosRESUMO
PURPOSE: To identify the association between several contextual match factors, technical performance, and external movement demands on the subjective task load of elite rugby league players. METHODS: Individual subjective task load, quantified using the National Aeronautics and Space Administration Task Load Index (NASA-TLX), was collected from 29 professional rugby league players from one club competing in the European Super League throughout the 2017 season. The sample consisted of 26 matches (441 individual data points). Linear mixed modeling revealed that various combinations of contextual factors, technical performance, and movement demands were associated with subjective task load. RESULTS: Greater number of tackles (effect size correlation ± 90% confidence intervals; η2 = .18 ± .11), errors (η2 = .15 ± .08), decelerations (η2 = .12 ± .08), increased sprint distance (η2 = .13 ± .08), losing matches (η2 = .36 ± .08), and increased perception of effort (η2 = .27 ± .08) led to most likely-very likely increases in subjective total task load. The independent variables included in the final model for subjective mental demand (match outcome, time played, and number of accelerations) were unclear, excluding a likely small correlation with technical errors (η2 = .10 ± .08). CONCLUSIONS: These data provide a greater understanding of the subjective task load and their association with several contextual factors, technical performance, and external movement demands during rugby league competition. Practitioners could use this detailed quantification of internal loads to inform recovery sessions and current training practices.
Assuntos
Desempenho Atlético , Futebol Americano , Corrida , Aceleração , Sistemas de Informação Geográfica , Humanos , MovimentoRESUMO
PURPOSE: To examine responses to a simulated rugby league protocol designed to include more stochastic commands, and therefore require greater vigilance, than traditional team-sport simulation protocols. METHODS: Eleven male university rugby players completed 2 trials (randomized and control [CON]) of a rugby league movement simulation protocol, separated by 7 to 10 d. The CON trial consisted of 48 repeated â¼115-s cycles of activity. The stochastic simulation (STOCH) was matched for the number and types of activity performed every 5.45 min in CON but included no repeated cycles of activity. Movement using GPS, heart rate, rating of perceived exertion, and Stroop test performance was assessed throughout. Maximum voluntary contraction peak torque, voluntary activation (in percentage), and global task load were assessed after exercise. RESULTS: The mean mental demand of STOCH was higher than CON (effect size [ES] = 0.56; ±0.69). Mean sprint speed was higher in STOCH (22.5 [1.4] vs 21.6 [1.6] km·h-1, ES = 0.50; ±0.55), which was accompanied by a higher rating of perceived exertion (14.3 [1.0] vs 13.0 [1.4], ES = 0.87; ±0.67) and a greater number of errors in the Stroop test (10.3 [2.5] vs 9.3 [1.4] errors; ES = 0.65; ±0.83). Maximum voluntary contraction peak torque (CON = -48.4 [31.6] N·m and STOCH = -39.6 [36.6] N·m) and voluntary activation (CON = -8.3% [4.8%] and STOCH = -6.0% [4.1%]) was similarly reduced in both trials. CONCLUSIONS: Providing more stochastic commands, which requires greater vigilance, might alter performance and associated physiological, perceptual, and cognitive responses to team-sport simulations.
Assuntos
Desempenho Atlético/fisiologia , Desempenho Atlético/psicologia , Futebol Americano/fisiologia , Futebol Americano/psicologia , Frequência Cardíaca , Humanos , Masculino , Movimento , Contração Muscular , Esforço Físico , Corrida , Teste de StroopRESUMO
PURPOSE: Several hundred genetic muscle diseases have been described, all of which are rare. Their clinical and genetic heterogeneity means that a genetic diagnosis is challenging. We established an international consortium, MYO-SEQ, to aid the work-ups of muscle disease patients and to better understand disease etiology. METHODS: Exome sequencing was applied to 1001 undiagnosed patients recruited from more than 40 neuromuscular disease referral centers; standardized phenotypic information was collected for each patient. Exomes were examined for variants in 429 genes associated with muscle conditions. RESULTS: We identified suspected pathogenic variants in 52% of patients across 87 genes. We detected 401 novel variants, 116 of which were recurrent. Variants in CAPN3, DYSF, ANO5, DMD, RYR1, TTN, COL6A2, and SGCA collectively accounted for over half of the solved cases; while variants in newer disease genes, such as BVES and POGLUT1, were also found. The remaining well-characterized unsolved patients (48%) need further investigation. CONCLUSION: Using our unique infrastructure, we developed a pathway to expedite muscle disease diagnoses. Our data suggest that exome sequencing should be used for pathogenic variant detection in patients with suspected genetic muscle diseases, focusing first on the most common disease genes described here, and subsequently in rarer and newly characterized disease genes.
Assuntos
Exoma , Distrofia Muscular do Cíngulo dos Membros , Anoctaminas , Exoma/genética , Glucosiltransferases , Humanos , Distrofia Muscular do Cíngulo dos Membros/genética , Sequenciamento do ExomaRESUMO
OBJECTIVE: To determine the proportion of infant deaths occurring in the setting of a confirmed genetic disorder. STUDY DESIGN: A retrospective analysis of the electronic medical records of infants born from 1 January, 2011 to 1 June, 2017, who died prior to 1 year of age. RESULTS: Five hundred and seventy three deceased infants were identified. One hundred and seventeen were confirmed to have a molecular or cytogenetic diagnosis in a clinical diagnostic laboratory and an additional seven were diagnosed by research testing for a total of 124/573 (22%) diagnosed infants. A total of 67/124 (54%) had chromosomal disorders and 58/124 (47%) had single gene disorders (one infant had both). The proportion of diagnoses made by sequencing technologies, such as exome sequencing, increased over the years. CONCLUSIONS: The prevalence of confirmed genetic disorders within our cohort of infant deaths is higher than that previously reported. Increased efforts are needed to further understand the mortality burden of genetic disorders in infancy.
Assuntos
Doenças Genéticas Inatas/mortalidade , Mortalidade Infantil , Transtornos Cromossômicos/epidemiologia , Transtornos Cromossômicos/mortalidade , Feminino , Doenças Genéticas Inatas/epidemiologia , Humanos , Lactente , Morte do Lactente/etiologia , Recém-Nascido , Masculino , Prevalência , Estudos Retrospectivos , Estados Unidos/epidemiologiaRESUMO
The study assesses the test-retest reliability of movement and physiological measures during a simulated rugby match that employed activities performed in a stochastic order. Twenty male rugby players (21.4 ± 2.1 y) completed two trials of a 2 × 23 min rugby movement simulation protocol during which the order of events was performed in a stochastic order, with 7-10 days between trials. Movement characteristics, heart rate (HR), RPE, maximum voluntary contraction (MVC), voluntary activation (VA%) of the quadriceps, Stroop test and subjective task load rating (NASA-TLX) were measured. The most reliable measures of external load was relative distance (typical error [TE] and CV% = 1.5-1.6 m min-1 and 1.4-1.5%, respectively), with all other movement characteristics possessing a CV% <5%. The most reliable measure of internal load, neuromuscular function and perceptual measures were for %HRmax (TE and CV% = 1.4-1.7% and 1.4-2.1%, respectively), MVC before (TE and CV% = 10.8-14.8 N·m and 3.8-4.6%, respectively), and average RPE (TE and CV% = 0.5-0.8 AU and 3.6-5.5%, respectively). The Stroop test, NASA-TLX and blood lactate produced the least reliable measures (CV% >5%). Future studies can confidently examine changes in several perceptual, neuromuscular, physiological and movement measures related to rugby activity using stochastic movements.
Assuntos
Desempenho Atlético/fisiologia , Desempenho Atlético/psicologia , Cognição/fisiologia , Futebol Americano/fisiologia , Futebol Americano/psicologia , Movimento/fisiologia , Músculo Quadríceps/fisiologia , Comportamento Competitivo/fisiologia , Frequência Cardíaca/fisiologia , Humanos , Ácido Láctico/sangue , Masculino , Contração Muscular/fisiologia , Percepção/fisiologia , Esforço Físico/fisiologia , Processos Estocásticos , Teste de Stroop , Análise e Desempenho de Tarefas , Adulto JovemRESUMO
BACKGROUND: Dystroglycanopathies are a clinically and genetically heterogeneous group of disorders that are typically characterised by limb-girdle muscle weakness. Mutations in 18 different genes have been associated with dystroglycanopathies, the encoded proteins of which typically modulate the binding of α-dystroglycan to extracellular matrix ligands by altering its glycosylation. This results in a disruption of the structural integrity of the myocyte, ultimately leading to muscle degeneration. METHODS: Deep phenotypic information was gathered using the PhenoTips online software for 1001 patients with unexplained limb-girdle muscle weakness from 43 different centres across 21 European and Middle Eastern countries. Whole-exome sequencing with at least 250 ng DNA was completed using an Illumina exome capture and a 38 Mb baited target. Genes known to be associated with dystroglycanopathies were analysed for disease-causing variants. RESULTS: Suspected pathogenic variants were detected in DPM3, ISPD, POMT1 and FKTN in one patient each, in POMK in two patients, in GMPPB in three patients, in FKRP in eight patients and in POMT2 in ten patients. This indicated a frequency of 2.7% for the disease group within the cohort of 1001 patients with unexplained limb-girdle muscle weakness. The phenotypes of the 27 patients were highly variable, yet with a fundamental presentation of proximal muscle weakness and elevated serum creatine kinase. CONCLUSIONS: Overall, we have identified 27 patients with suspected pathogenic variants in dystroglycanopathy-associated genes. We present evidence for the genetic and phenotypic diversity of the dystroglycanopathies as a disease group, while also highlighting the advantage of incorporating next-generation sequencing into the diagnostic pathway of rare diseases.
Assuntos
Variação Genética , Distrofia Muscular do Cíngulo dos Membros/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Distroglicanas/metabolismo , Feminino , Predisposição Genética para Doença , Glicosilação , Heterozigoto , Homozigoto , Humanos , Masculino , Pessoa de Meia-Idade , Proteínas Musculares/genética , Distrofia Muscular do Cíngulo dos Membros/metabolismo , Mutação , Fenótipo , Sequenciamento do Exoma/métodos , Adulto JovemRESUMO
The transcription factor BCL11B is essential for development of the nervous and the immune system, and Bcl11b deficiency results in structural brain defects, reduced learning capacity, and impaired immune cell development in mice. However, the precise role of BCL11B in humans is largely unexplored, except for a single patient with a BCL11B missense mutation, affected by multisystem anomalies and profound immune deficiency. Using massively parallel sequencing we identified 13 patients bearing heterozygous germline alterations in BCL11B. Notably, all of them are affected by global developmental delay with speech impairment and intellectual disability; however, none displayed overt clinical signs of immune deficiency. Six frameshift mutations, two nonsense mutations, one missense mutation, and two chromosomal rearrangements resulting in diminished BCL11B expression, arose de novo. A further frameshift mutation was transmitted from a similarly affected mother. Interestingly, the most severely affected patient harbours a missense mutation within a zinc-finger domain of BCL11B, probably affecting the DNA-binding structural interface, similar to the recently published patient. Furthermore, the most C-terminally located premature termination codon mutation fails to rescue the progenitor cell proliferation defect in hippocampal slice cultures from Bcl11b-deficient mice. Concerning the role of BCL11B in the immune system, extensive immune phenotyping of our patients revealed alterations in the T cell compartment and lack of peripheral type 2 innate lymphoid cells (ILC2s), consistent with the findings described in Bcl11b-deficient mice. Unsupervised analysis of 102 T lymphocyte subpopulations showed that the patients clearly cluster apart from healthy children, further supporting the common aetiology of the disorder. Taken together, we show here that mutations leading either to BCL11B haploinsufficiency or to a truncated BCL11B protein clinically cause a non-syndromic neurodevelopmental delay. In addition, we suggest that missense mutations affecting specific sites within zinc-finger domains might result in distinct and more severe clinical outcomes.
Assuntos
Transtornos do Neurodesenvolvimento/genética , Proteínas Repressoras/genética , Proteínas Repressoras/fisiologia , Proteínas Supressoras de Tumor/genética , Proteínas Supressoras de Tumor/fisiologia , Adolescente , Animais , Criança , Pré-Escolar , Feminino , Regulação da Expressão Gênica/genética , Mutação em Linhagem Germinativa , Haploinsuficiência , Heterozigoto , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Lactente , Linfócitos/patologia , Linfócitos/fisiologia , Masculino , Camundongos , Mutação , Proteínas Repressoras/metabolismo , Linfócitos T/fisiologia , Fatores de Transcrição/genética , Proteínas Supressoras de Tumor/metabolismoRESUMO
The development of methods to produce nanoscale features with tailored chemical functionalities is fundamental for applications such as nanoelectronics and sensor fabrication. The molecular-ruler process shows great utility for this purpose as it combines top-down lithography for the creation of complex architectures over large areas in conjunction with molecular self-assembly, which enables precise control over the physical and chemical properties of small local features. The molecular-ruler process, which most commonly uses mercaptoalkanoic acids and metal ions to generate metal-ligated multilayers, can be employed to produce registered nanogaps between metal features. Expansion of this methodology to include molecules with other chemical functionalities could greatly expand the overall versatility, and thus the utility, of this process. Herein, we explore the use of alkanethiol molecules as the terminating layer of metal-ligated multilayers. During this study, it was discovered that the solution deposition of alkanethiol molecules resulted in low overall surface coverage with features that varied in height. Because features with varied heights are not conducive to the production of uniform nanogaps via the molecular-ruler process, the vapor-phase deposition of alkanethiol molecules was explored. Unlike the solution-phase deposition, alkanethiol islands produced by vapor-phase deposition exhibited markedly higher surface coverages of uniform heights. To illustrate the applicability of this method, metal-ligated multilayers, both with and without an alkanethiol capping layer, were utilized to create nanogaps between Au features using the molecular-ruler process.
RESUMO
Ribosomal RNA (rRNA) is transcribed from rDNA by RNA polymerase I (Pol I) to produce the 45S precursor of the 28S, 5.8S, and 18S rRNA components of the ribosome. Two transcription factors have been defined for Pol I in mammals, the selectivity factor SL1, and the upstream binding transcription factor (UBF), which interacts with the upstream control element to facilitate the assembly of the transcription initiation complex including SL1 and Pol I. In seven unrelated affected individuals, all suffering from developmental regression starting at 2.5-7 years, we identified a heterozygous variant, c.628G>A in UBTF, encoding p.Glu210Lys in UBF, which occurred de novo in all cases. While the levels of UBF, Ser388 phosphorylated UBF, and other Pol I-related components (POLR1E, TAF1A, and TAF1C) remained unchanged in cells of an affected individual, the variant conferred gain of function to UBF, manifesting by markedly increased UBF binding to the rDNA promoter and to the 5'- external transcribed spacer. This was associated with significantly increased 18S expression, and enlarged nucleoli which were reduced in number per cell. The data link neurodegeneration in childhood with altered rDNA chromatin status and rRNA metabolism.
Assuntos
Encefalopatias/genética , Nucléolo Celular/patologia , Doenças Neurodegenerativas/genética , Proteínas Pol1 do Complexo de Iniciação de Transcrição/genética , RNA Ribossômico 18S/biossíntese , Adolescente , Adulto , Atrofia/genética , Encéfalo/patologia , Encefalopatias/patologia , Criança , Cromatina/metabolismo , Proteínas de Ligação a DNA/genética , Feminino , Humanos , Masculino , Doenças Neurodegenerativas/patologia , Polimorfismo de Nucleotídeo Único/genética , Regiões Promotoras Genéticas/genética , Adulto JovemRESUMO
PURPOSE: To examine the influence of knowledge of exercise duration on pacing and performance during simulated rugby league match play. METHODS: Thirteen male university rugby players completed 3 simulated rugby league matches (RLMSP-i) on separate days in a random order. In a control trial, participants were informed that they would be performing 2 × 23-min bouts (separated by 20 min) of the RLMSP-i (CON). In a second trial, participants were informed that they would be performing 1 × 23-min bout of the protocol but were then asked to perform another 23-min bout (DEC). In a third trial, participants were not informed of the exercise duration and performed 2 × 23-min bouts (UN). RESULTS: Distance covered and high-intensity running were higher in CON (4813 ± 167 m, 26 ± 4.1 m/min) than DEC (4764 ± 112 m, 25.2 ± 2.8 m/min) and UN (4744 ± 131 m, 24.4 m/min). Compared with CON, high-intensity running and peak speed were typically higher for DEC in bout 1 and lower in bout 2 of the RLMSP-i, while UN was generally lower throughout. Similarly, DEC resulted in an increased heart rate, blood lactate, and rating of perceived exertion than CON in bout 1, whereas these variables were lower throughout the protocol in UN. CONCLUSIONS: Pacing and performance during simulated rugby league match play depend on an accurate understanding of the exercise endpoint. Applied practitioners should consider informing players of their likely exercise duration to maximize running.
Assuntos
Desempenho Atlético/psicologia , Futebol Americano/psicologia , Esforço Físico , Adulto , Desempenho Atlético/fisiologia , Futebol Americano/fisiologia , Frequência Cardíaca , Humanos , Ácido Láctico/sangue , Masculino , Corrida/fisiologia , Corrida/psicologia , Fatores de Tempo , Adulto JovemRESUMO
Gain-of-function variants in some RAS-MAPK pathway genes, including PTPN11 and NRAS, are associated with RASopathies and/or acquired hematological malignancies, most notably juvenile myelomonocytic leukemia (JMML). With rare exceptions, the spectrum of germline variants causing RASopathies does not overlap with the somatic variants identified in isolated JMML. Studies comparing these variants suggest a stronger gain-of-function activity in the JMML variants. As JMML variants have not been identified as germline defects and have a greater impact on protein function, it has been speculated that they would be embryonic lethal. Here we identified three variants, which have previously only been identified in isolated somatic JMML and other sporadic cancers, in four cases with a severe pre- or neo-natal lethal presentation of Noonan syndrome. These cases support the hypothesis that these stronger gain-of-function variants are rarely compatible with life.
Assuntos
GTP Fosfo-Hidrolases/genética , Mutação em Linhagem Germinativa , Leucemia Mielomonocítica Juvenil/genética , Proteínas de Membrana/genética , Síndrome de Noonan/genética , Proteína Tirosina Fosfatase não Receptora Tipo 11/genética , Feminino , Humanos , Recém-Nascido , Síndrome de Noonan/diagnóstico , GravidezRESUMO
This aim of this study was to examine the validity of energy expenditure derived from microtechnology when measured during a repeated-effort rugby protocol. Sixteen male rugby players completed a repeated-effort protocol comprising 3 sets of 6 collisions during which movement activity and energy expenditure (EEGPS) were measured using microtechnology. In addition, energy expenditure was estimated from open-circuit spirometry (EEVO2). While related (r = .63, 90%CI .08-.89), there was a systematic underestimation of energy expenditure during the protocol (-5.94 ± 0.67 kcal/min) for EEGPS (7.2 ± 1.0 kcal/min) compared with EEVO2 (13.2 ± 2.3 kcal/min). High-speed-running distance (r = .50, 95%CI -.66 to .84) was related to EEVO2, while PlayerLoad was not (r = .37, 95%CI -.81 to .68). While metabolic power might provide a different measure of external load than other typically used microtechnology metrics (eg, high-speed running, PlayerLoad), it underestimates energy expenditure during intermittent team sports that involve collisions.
Assuntos
Acelerometria , Metabolismo Energético , Futebol Americano/fisiologia , Sistemas de Informação Geográfica , Frequência Cardíaca/fisiologia , Humanos , Ácido Láctico/sangue , Masculino , Percepção , Condicionamento Físico Humano , Esforço Físico/fisiologia , Corrida/fisiologia , Adulto JovemRESUMO
PURPOSE: Tools for assessing the severity and risk of near-miss events in radiation oncology are few and needed. Recent work has described guidelines for the use of a 5-tier near-miss risk index (NMRI) for the classification of near-miss events. The purpose of this study was to assess the reliability of the NMRI among users in a radiation oncology department. METHODS AND MATERIALS: Reliability of the NMRI was assessed using an online survey distributed to members of a radiation oncology department. The survey contained 70 events extracted from the department's incident learning system (ILS). Survey participants rated each event using the NMRI guidelines, reported their attendance to weekly ILS meetings (used as a surrogate for familiarity with the ILS), and indicated their familiarity with the radiation oncology workflow. Interrater reliability was determined using Krippendorff's alpha. Use of the NMRI to rate actual events during 5 weekly ILS meetings was also assessed and interrater reliability determined. RESULTS: Twenty-eight survey respondents represented a wide variety of care providers. Krippendorff's alpha was calculated for the whole respondent cohort to be 0.376, indicating fair agreement among raters. Respondents who had the most participation at ILS meetings (n = 4) had moderate agreement with an alpha of 0.501. Interestingly, there were significant differences in reliability and median NMRI scores between professions. NMRI use during weekly NMRI meetings (80 events rated), participants showed moderate reliability (alpha = 0.607). CONCLUSIONS: Using the NMRI guidelines, raters from a wide variety of professions were able to assess the severity of near-miss incidents with fair agreement. Those experienced with the ILS showed better agreement, and higher agreement was seen during multidisciplinary ILS meetings. These data support the use the indices such as the NMRI for near-miss risk assessment in patient safety and prioritization of process improvements in radiation oncology.
Assuntos
Near Miss/estatística & dados numéricos , Variações Dependentes do Observador , Radioterapia (Especialidade) , Medição de Risco/métodos , Humanos , Reprodutibilidade dos Testes , Inquéritos e Questionários , Fluxo de TrabalhoRESUMO
PURPOSE: There is a growing interest in the application of incident learning systems (ILS) to radiation oncology. The purpose of the present study is to define statistical metrics that may serve as benchmarks for successful operation of an incident learning system. METHODS AND MATERIALS: A departmental safety and quality ILS was developed to monitor errors, near-miss events, and process improvement suggestions. Event reports were reviewed by a multiprofessional quality improvement committee. Events were scored by a near-miss risk index (NMRI) and categorized by event point of origination and discovery. Reporting trends were analyzed over a 2-year period, including total number and rates of events reported, users reporting, NMRI, and event origination and discovery. RESULTS: A total of 1897 reports were evaluated (1.0 reports/patient, 0.9 reports/unique treatment course). Participation in the ILS increased as demonstrated by total events (2.1 additional reports/month) and unique users (0.5 new users/month). Sixteen percent of reports had an NMRI of 0 (none), 42% had an NMRI of 1 (mild), 25% had an NMRI of 2 (moderate), 12% had an NMRI of 3 (severe), and 5% had an NMRI of 4 (critical). Event NMRI showed a significant decrease in the first 6 months (1.68-1.42, P < .001). Trends in origination and discovery of reports were broadly distributed between radiation therapy process steps and staff groups. The highest risk events originated in imaging for treatment planning (NMRI = 2.0 ± 1.1; P < .0001) and were detected in on-treatment quality management (NMRI = 1.7 ± 1.1; P = .003). CONCLUSIONS: Over the initial 2-year period of ILS operation, rates of reporting increased, staff participation increased, and NMRI of reported events declined. These data mirror previously reported findings of improvement in safety culture endpoints. These metrics may be useful for other institutions seeking to create or evaluate their own ILS.