RESUMO
To execute accurate movements, animals must continuously adapt their behavior to changes in their bodies and environments. Animals can learn changes in the relationship between their locomotor commands and the resulting distance moved, then adjust command strength to achieve a desired travel distance. It is largely unknown which circuits implement this form of motor learning, or how. Using whole-brain neuronal imaging and circuit manipulations in larval zebrafish, we discovered that the serotonergic dorsal raphe nucleus (DRN) mediates short-term locomotor learning. Serotonergic DRN neurons respond phasically to swim-induced visual motion, but little to motion that is not self-generated. During prolonged exposure to a given motosensory gain, persistent DRN activity emerges that stores the learned efficacy of motor commands and adapts future locomotor drive for tens of seconds. The DRN's ability to track the effectiveness of motor intent may constitute a computational building block for the broader functions of the serotonergic system. VIDEO ABSTRACT.
Assuntos
Aprendizagem , Modelos Neurológicos , Natação , Peixe-Zebra/fisiologia , Animais , Mapeamento Encefálico , Larva , Optogenética , Núcleos da Rafe/fisiologia , Neurônios Serotoninérgicos/citologia , Neurônios Serotoninérgicos/fisiologia , Processamento EspacialRESUMO
The Arc/Arg3.1 gene product is rapidly upregulated by strong synaptic activity and critically contributes to weakening synapses by promoting AMPA-R endocytosis. However, how activity-induced Arc is redistributed and determines the synapses to be weakened remains unclear. Here, we show targeting of Arc to inactive synapses via a high-affinity interaction with CaMKIIß that is not bound to calmodulin. Synaptic Arc accumulates in inactive synapses that previously experienced strong activation and correlates with removal of surface GluA1 from individual synapses. A lack of CaMKIIß either in vitro or in vivo resulted in loss of Arc upregulation in the silenced synapses. The discovery of Arc's role in "inverse" synaptic tagging that is specific for weaker synapses and prevents undesired enhancement of weak synapses in potentiated neurons reconciles essential roles of Arc both for the late phase of long-term plasticity and for reduction of surface AMPA-Rs in stimulated neurons.
Assuntos
Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/metabolismo , Proteínas do Citoesqueleto/metabolismo , Hipocampo/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Neurônios/metabolismo , Sinapses/metabolismo , Animais , Células Cultivadas , Espinhas Dendríticas/metabolismo , Hipocampo/citologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Ratos , Ratos Sprague-DawleyRESUMO
Serotonergic psychedelics are emerging therapeutics for psychiatric disorders, yet their underlying mechanisms of action in the brain remain largely elusive. Here, we developed a wide-field behavioral tracking system for larval zebrafish and investigated the effects of psilocybin, a psychedelic serotonin receptor agonist. Machine learning analyses of precise body kinematics identified latent behavioral states reflecting spontaneous exploration, visually-driven rapid swimming, and irregular swim patterns following stress exposure. Using this method, we found that acute psilocybin treatment has two behavioral effects: [i] facilitation of spontaneous exploration ("stimulatory") and [ii] prevention of irregular swim patterns following stress exposure ("anxiolytic"). These effects differed from the effect of acute SSRI treatment and were rather similar to the effect of ketamine treatment. Neural activity imaging in the dorsal raphe nucleus suggested that psilocybin inhibits serotonergic neurons by activating local GABAergic neurons, consistent with psychedelic-induced suppression of serotonergic neurons in mammals. These findings pave the way for using larval zebrafish to elucidate neural mechanisms underlying the behavioral effects of serotonergic psychedelics.
Assuntos
Ansiolíticos , Comportamento Animal , Alucinógenos , Psilocibina , Neurônios Serotoninérgicos , Peixe-Zebra , Animais , Psilocibina/farmacologia , Comportamento Animal/efeitos dos fármacos , Alucinógenos/farmacologia , Ansiolíticos/farmacologia , Neurônios Serotoninérgicos/efeitos dos fármacos , Larva/efeitos dos fármacos , Natação , Núcleo Dorsal da Rafe/efeitos dos fármacos , Agonistas do Receptor de Serotonina/farmacologia , Ansiedade/tratamento farmacológico , Neurônios GABAérgicos/efeitos dos fármacos , Inibidores Seletivos de Recaptação de Serotonina/farmacologia , Ketamina/farmacologia , Encéfalo/efeitos dos fármacosRESUMO
BACKGROUND: This study assessed the differences in daily virus reduction and the residual infectivity after the recommended home stay period in Japan in patients infected with influenza and treated with baloxavir (BA), laninamivir (LA), oseltamivir (OS), and zanamivir (ZA). METHODS: We conducted an observational study on children and adults at 13 outpatient clinics in 11 prefectures in Japan during seven influenza seasons from 2013/2014 to 2019/2020. Virus samples were collected twice from influenza rapid test-positive patients at the first and second visit 4-5 days after the start of treatment. The viral RNA shedding was quantified using quantitative RT-PCR. Neuraminidase (NA) and polymerase acidic (PA) variant viruses that reduce susceptibility to NA inhibitors and BA, respectively, were screened using RT-PCR and genetic sequencing. Daily estimated viral reduction was evaluated using univariate and multivariate analyses for the factors such as age, treatment, vaccination status, or the emergence of PA or NA variants. The potential infectivity of the viral RNA shedding at the second visit samples was determined using the Receiver Operator Curve based on the positivity of virus isolation. RESULTS: Among 518 patients, 465 (80.0%) and 116 (20.0%) were infected with influenza A (189 with BA, 58 with LA, 181 with OS, 37 with ZA) and influenza B (39 with BA, 10 with LA, 52 with OS, 15 with ZA). The emergence of 21 PA variants in influenza A was detected after BA treatment, but NA variants were not detected after NAIs treatment. Multiple linear regression analysis showed that the daily viral RNA shedding reduction in patients was slower in the two NAIs (OS and LA) than in BA, influenza B infection, aged 0-5 years, or the emergence of PA variants. The residual viral RNA shedding potentially infectious was detected in approximately 10-30% of the patients aged 6-18 years after five days of onset. CONCLUSIONS: Viral clearance differed by age, type of influenza, choice of treatment, and susceptibility to BA. Additionally, the recommended homestay period in Japan seemed insufficient, but reduced viral spread to some extent since most school-age patients became non-infectious after 5 days of onset.
Assuntos
Influenza Humana , Criança , Adulto , Humanos , Influenza Humana/tratamento farmacológico , Neuraminidase/genética , Pacientes Ambulatoriais , Japão , Estações do Ano , Antivirais/uso terapêutico , Antivirais/farmacologia , Zanamivir/uso terapêutico , Oseltamivir/uso terapêutico , Inibidores Enzimáticos/uso terapêutico , RNA Viral/genéticaRESUMO
Current techniques for monitoring GABA (γ-aminobutyric acid), the primary inhibitory neurotransmitter in vertebrates, cannot follow transients in intact neural circuits. To develop a GABA sensor, we applied the design principles used to create the fluorescent glutamate receptor iGluSnFR. We used a protein derived from a previously unsequenced Pseudomonas fluorescens strain and performed structure-guided mutagenesis and library screening to obtain intensity-based GABA sensing fluorescence reporter (iGABASnFR) variants. iGABASnFR is genetically encoded, detects GABA release evoked by electric stimulation of afferent fibers in acute brain slices and produces readily detectable fluorescence increases in vivo in mice and zebrafish. We applied iGABASnFR to track mitochondrial GABA content and its modulation by an anticonvulsant, swimming-evoked, GABA-mediated transmission in zebrafish cerebellum, GABA release events during interictal spikes and seizures in awake mice, and found that GABA-mediated tone decreases during isoflurane anesthesia.
Assuntos
Técnicas Biossensoriais/métodos , Encéfalo/metabolismo , Proteínas de Fluorescência Verde/metabolismo , Hipocampo/metabolismo , Imagem Molecular/métodos , Neurônios/metabolismo , Ácido gama-Aminobutírico/metabolismo , Anestesia , Animais , Animais Geneticamente Modificados , Feminino , Proteínas de Fluorescência Verde/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Ratos , Ratos Sprague-Dawley , Convulsões/metabolismo , Convulsões/patologia , Peixe-ZebraRESUMO
In the version of this article originally published, the bottom of Figure 4f,g was partially truncated in the PDF. The error has been corrected in the PDF version of this article.
RESUMO
We developed a new way to engineer complex proteins toward multidimensional specifications using a simple, yet scalable, directed evolution strategy. By robotically picking mammalian cells that were identified, under a microscope, as expressing proteins that simultaneously exhibit several specific properties, we can screen hundreds of thousands of proteins in a library in just a few hours, evaluating each along multiple performance axes. To demonstrate the power of this approach, we created a genetically encoded fluorescent voltage indicator, simultaneously optimizing its brightness and membrane localization using our microscopy-guided cell-picking strategy. We produced the high-performance opsin-based fluorescent voltage reporter Archon1 and demonstrated its utility by imaging spiking and millivolt-scale subthreshold and synaptic activity in acute mouse brain slices and in larval zebrafish in vivo. We also measured postsynaptic responses downstream of optogenetically controlled neurons in C. elegans.
Assuntos
Evolução Molecular Direcionada/métodos , Proteínas Luminescentes/química , Engenharia de Proteínas/métodos , Robótica , Peixe-Zebra/embriologia , Animais , Encéfalo/diagnóstico por imagem , Caenorhabditis elegans , Separação Celular , Feminino , Citometria de Fluxo , Fluorescência , Biblioteca Gênica , Genes Reporter , Células HEK293 , Hipocampo/citologia , Humanos , Masculino , Camundongos , Microscopia de Fluorescência , Neurônios/citologia , OptogenéticaRESUMO
An aberrant right subclavian artery (ARSCA) is a relatively rare congenital anomaly of the aortic arch branches. A 63-year-old man suffered from dysphagia, and was referred to our hospital. Computed tomography (CT) revealed an aortic aneurysm (Kommerell's diverticulum) and ARSCA which routed behind the esophagus. We performed total arch replacement with the open stent-grafting technique via median sternotomy. ARSCA was anastomosed to 1 branch of the arch graft at the right side of the trachea, which released esophageal compression. Postoperatively dysphagia disappeared and CT scan indicated successful reconstruction of the distal arch and ARSCA. The open stent-grafting technique is considered to be effective for aortic disease with ARSCA.
Assuntos
Anormalidades Cardiovasculares/cirurgia , Transtornos de Deglutição , Artéria Subclávia/anormalidades , Aorta Torácica , Anormalidades Cardiovasculares/complicações , Transtornos de Deglutição/etiologia , Divertículo , Humanos , Masculino , Pessoa de Meia-Idade , Stents , Artéria Subclávia/cirurgiaRESUMO
A 67-year-old woman was admitted with exertional dyspnea. She had undergone mitral valve replacement with Björk-Shiley prosthetic valve (convexo-concave type) 36 years previously for stenosis and regurgitation in another institute. An echocardiography showed a severe degree of aortic stenosis, and the implanted mitral valve function is normal. We performed aortic valve replacement with a bioprosthetic valve and no prophylactic reoperation for the implanted mitral valve. The postoperative courses were uneventful. The patient was discharged from the hospital in good clinical condition. Postoperative echocardiography showed the prosthetic valves were normal. The close follow-up should be needed to prevent fatal valve dysfunction.
Assuntos
Valva Aórtica , Próteses Valvulares Cardíacas , Idoso , Feminino , Humanos , Valva Mitral , Falha de Prótese , ReoperaçãoRESUMO
Right-sided aortic arch (RAA) is a rare congenital disorder. A 87-year-old man with chest pain was diagnosed with a right-sided aortic arch and Kommerell's diverticulum by computed tomography (CT). The diverticulum had a maximum diameter of 57 mm, and surgical intervention was chosen because of the possibility of rupture and dissection. We performed thoracic endovascular aortic repair( TEVAR) with a commercially available device, consisting of bypass grafting of the supra-aortic branches. The postoperative courses were uneventful. The patient was discharged from the hospital in good clinical condition. Postoperative CT showed complete exclusion of the diverticulum without endoleak and device migration. Debranching TEVAR for Kommerell's diverticulum was safe and effective.
Assuntos
Divertículo , Procedimentos Endovasculares , Idoso de 80 Anos ou mais , Aorta Torácica , Humanos , Masculino , Artéria Subclávia , Tomografia Computadorizada por Raios XRESUMO
The reactions of the diruthenium tetrahydrido complex, Cp*Ru(µ-H)4RuCp* (1) (Cp* = η5-C5Me5), with pyridines were investigated in relation to the dehydrogenative coupling of 4-substituted pyridines. Complex 1 reacted with γ-picoline to yield the bis(µ-pyridyl) complex, {Cp*Ru(µ-H)(µ-4-MeC5H3)}2 (2a), with the elimination of dihydrogen. Complex 2a immediately reacted with the liberated dihydrogen to yield µ-η2-dihydrobipyridine (dhbpy) complex 4avia C-C bond formation between the two pyridyl groups, in which one of the pyridine rings underwent partial hydrogenation. The X-ray structure of 4a shows that the dhbpy moiety adopts a µ-η2 coordination mode at the Ru2 site. Complex 4a was reversibly converted to 5avia the elimination of dihydrogen in which the dhbpy moiety adopts a µ-η2:η2 mode. Although 5a was coordinatively saturated, 5a readily reacted with tBuNC to yield 6a. This was owing to the ability of the dhbpy ligand changing its coordination mode between the µ-η2:η2 and µ-η2 modes. This also causes the dehydrogenation from the dhbpy ligand to yield µ-η2:η2-bipyridine complex 7a at 140 °C. However, 7a was not shown to be an intermediate of the catalysis. The reaction of 1 with 1,10-phenanthroline afforded µ-η2-phenanthroline complex 8 containing two hydrides, which can be a model compound for the bipyridine elimination from the Ru2 site. Dynamic NMR studies suggested that 8 was isomerised to an unsaturated µ-N-heterocyclic carbene (NHC) complex. The unsaturated nature of the µ-NHC complex is likely responsible for the uptake of the third pyridine molecule to turn over the catalytic cycle.
RESUMO
Understanding brain function requires monitoring and interpreting the activity of large networks of neurons during behavior. Advances in recording technology are greatly increasing the size and complexity of neural data. Analyzing such data will pose a fundamental bottleneck for neuroscience. We present a library of analytical tools called Thunder built on the open-source Apache Spark platform for large-scale distributed computing. The library implements a variety of univariate and multivariate analyses with a modular, extendable structure well-suited to interactive exploration and analysis development. We demonstrate how these analyses find structure in large-scale neural data, including whole-brain light-sheet imaging data from fictively behaving larval zebrafish, and two-photon imaging data from behaving mouse. The analyses relate neuronal responses to sensory input and behavior, run in minutes or less and can be used on a private cluster or in the cloud. Our open-source framework thus holds promise for turning brain activity mapping efforts into biological insights.
Assuntos
Potenciais de Ação/fisiologia , Mapeamento Encefálico/métodos , Armazenamento e Recuperação da Informação/métodos , Modelos Neurológicos , Rede Nervosa/fisiologia , Neurônios/fisiologia , Software , Animais , Encéfalo/fisiologia , Simulação por Computador , Metodologias Computacionais , Interpretação Estatística de Dados , Sistemas de Gerenciamento de Base de Dados , Bases de Dados Factuais , Humanos , Linguagens de ProgramaçãoRESUMO
Generally, infective endocarditis is found at the left side of the heart. The right side infective endocaritis accounts for only 5~10% of all cases of infective endocarditis. The right side infective endocarditis occurs especially among drug users employing intravenous injection. A typical site of infection is the tricupid valve, and isolated pulmonary valve involvement is rare. It is assumed that its rarity is due to the low pressure gradients within the right heart, the low prevalence of valve disease, and the lower oxygen content of the venous blood. We describe a case of isolated pulmonary valve endocarditis requiring valve replacement. Antibiotic therapy was conducted for 4 weeks before surgery and 2 weeks after surgery. Clinical course was favorable and the patent was discharged home 18 days after surgery.
Assuntos
Endocardite Bacteriana/cirurgia , Doenças das Valvas Cardíacas/cirurgia , Valva Pulmonar/cirurgia , Idoso , Procedimentos Cirúrgicos Cardíacos , Endocardite Bacteriana/complicações , Endocardite Bacteriana/diagnóstico por imagem , Doenças das Valvas Cardíacas/diagnóstico por imagem , Doenças das Valvas Cardíacas/etiologia , Humanos , Masculino , Streptococcus/isolamento & purificaçãoRESUMO
Identifying the neuronal ensembles that respond to specific stimuli and mapping their projection patterns in living animals are fundamental challenges in neuroscience. To this end, we engineered a synthetic promoter, the enhanced synaptic activity-responsive element (E-SARE), that drives neuronal activity-dependent gene expression more potently than other existing immediate-early gene promoters. Expression of a drug-inducible Cre recombinase downstream of E-SARE enabled imaging of neuronal populations that respond to monocular visual stimulation and tracking of their long-distance thalamocortical projections in living mice. Targeted cell-attached recordings and calcium imaging of neurons in sensory cortices revealed that E-SARE reporter expression correlates with sensory-evoked neuronal activity at the single-cell level and is highly specific to the type of stimuli presented to the animals. This activity-dependent promoter can expand the repertoire of genetic approaches for high-resolution anatomical and functional analysis of neural circuits.
Assuntos
Biologia Molecular/métodos , Neurônios/fisiologia , Regiões Promotoras Genéticas , Elementos de Resposta , Animais , Axônios , Cálcio/análise , Cálcio/metabolismo , Células Cultivadas , Dependovirus/genética , Feminino , Regulação da Expressão Gênica , Genes Reporter , Corpos Geniculados/citologia , Corpos Geniculados/fisiologia , Proteínas de Fluorescência Verde/análise , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Processamento de Imagem Assistida por Computador , Integrases/genética , Integrases/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Imagem Molecular/métodos , Estimulação Luminosa , Ratos , Ratos Sprague-Dawley , Análise de Célula Única/métodos , Córtex Visual/citologia , Córtex Visual/fisiologiaRESUMO
A 58-year-old man presented with severe aortic regurgitation(AR) with quadricuspid valve. Intraoperatively, the small accessory cusp was separated from non-coronary cusp( NCC). The NCC and small accessory cusp were sutured to obtain 1 competent cusp, aiming at an effective height of 8 mm. His AR was trivial at the postoperative 7th day.
Assuntos
Insuficiência da Valva Aórtica/cirurgia , Valva Aórtica/cirurgia , Anuloplastia da Valva Cardíaca/métodos , Humanos , Masculino , Pessoa de Meia-Idade , Período Pós-OperatórioRESUMO
We compared the duration of fever in children infected with A(H1N1)pdm09, A(H3N2), or influenza B viruses following treatment with baloxavir marboxil (baloxavir) or neuraminidase inhibitors (NAIs) (oseltamivir, zanamivir, or laninamivir). This observational study was conducted at 10 outpatient clinics across 9 prefectures in Japan during the 2012-2013 and 2019-2020 influenza seasons. Patients with influenza rapid antigen test positive were treated with one of four anti-influenza drugs. The type/subtype of influenza viruses were identified from MDCK or MDCK SIAT1 cell-grown samples using two-step real-time PCR. Daily self-reported body temperature after treatment were used to evaluate the duration of fever by treatment group and various underlying factors. Among 1742 patients <19 years old analyzed, 452 (26.0%) were A(H1N1)pdm09, 827 (48.0%) A(H3N2), and 463 (26.0%) influenza B virus infections. Among fours treatment groups, baloxavir showed a shorter median duration of fever compared to oseltamivir in univariate analysis for A(H1N1)pdm09 virus infections (baloxavir, 22.0 h versus oseltamivir, 26.7 h, P < 0.05; laninamivir, 25.5 h, and zanamivir, 25.0 h). However, this difference was not significant in multivariable analyses. For A(H3N2) virus infections, there were no statistically significant differences observed (20.3, 21.0, 22.0, and 19.0 h) uni- and multivariable analyses. For influenza B, baloxavir shortened the fever duration by approximately 15 h than NAIs (20.3, 35.0, 34.3, and 34.1 h), as supported by uni- and multivariable analyses. Baloxavir seems to have comparable clinical effectiveness with NAIs on influenza A but can be more effective for treating pediatric influenza B virus infections than NAIs.
Assuntos
Antivirais , Dibenzotiepinas , Febre , Guanidinas , Vírus da Influenza A Subtipo H1N1 , Vírus da Influenza A Subtipo H3N2 , Vírus da Influenza B , Influenza Humana , Morfolinas , Oseltamivir , Piranos , Piridonas , Ácidos Siálicos , Triazinas , Zanamivir , Humanos , Influenza Humana/tratamento farmacológico , Influenza Humana/virologia , Antivirais/uso terapêutico , Antivirais/farmacologia , Vírus da Influenza B/efeitos dos fármacos , Vírus da Influenza B/genética , Criança , Zanamivir/uso terapêutico , Zanamivir/análogos & derivados , Zanamivir/farmacologia , Triazinas/uso terapêutico , Triazinas/farmacologia , Guanidinas/uso terapêutico , Vírus da Influenza A Subtipo H3N2/efeitos dos fármacos , Vírus da Influenza A Subtipo H3N2/genética , Vírus da Influenza A Subtipo H1N1/efeitos dos fármacos , Piridonas/uso terapêutico , Dibenzotiepinas/uso terapêutico , Japão , Feminino , Masculino , Pré-Escolar , Oseltamivir/uso terapêutico , Febre/tratamento farmacológico , Febre/virologia , Adolescente , Morfolinas/uso terapêutico , Lactente , Estações do Ano , Tiepinas/uso terapêutico , Tiepinas/farmacologia , Oxazinas/uso terapêutico , Fatores de Tempo , Benzoxazinas/uso terapêuticoRESUMO
Baloxavir marboxil (baloxavir), approved as an anti-influenza drug in Japan in March 2018, can induce reduced therapeutic effectiveness due to PA protein substitutions. We assessed PA substitutions in clinical samples from influenza-infected children and adults pre- and post-baloxavir treatment, examining their impact on fever and symptom duration. During the 2022-2023 influenza season, the predominant circulating influenza subtype detected by cycling-probe RT-PCR was A(H3N2) (n = 234), with a minor circulation of A(H1N1)pdm09 (n = 10). Of the 234 influenza A(H3N2) viruses collected prior to baloxavir treatment, 2 (0.8%) viruses carry PA/I38T substitution. One virus was collected from a toddler and one from an adult, indicating the presence of viruses with reduced susceptibility to baloxavir, without prior exposure to the drug. Of the 54 paired influenza A(H3N2) viruses collected following baloxavir treatment, 8 (14.8%) viruses carried E23 K/G, or I38 M/T substitutions in PA. Variant calling through next-generation sequencing (NGS) showed varying proportions (6-100 %), a polymorphism and a mixture of PA/E23 K/G, and I38 M/T substitutions in the clinical samples. These eight viruses were obtained from children aged 7-14 years, with a median fever duration of 16.7 h and a median symptom duration of 93.7 h, which were similar to those of the wild type. However, the delayed viral clearance associated with the emergence of PA substitutions was observed. No substitutions conferring resistance to neuraminidase inhibitors were detected in 37 paired samples collected before and following oseltamivir treatment. These findings underscore the need for ongoing antiviral surveillance, informing public health strategies and clinical antiviral recommendations for seasonal influenza.
Assuntos
Substituição de Aminoácidos , Antivirais , Dibenzotiepinas , Farmacorresistência Viral , Vírus da Influenza A Subtipo H3N2 , Influenza Humana , Morfolinas , Piridonas , Triazinas , Proteínas Virais , Humanos , Dibenzotiepinas/uso terapêutico , Dibenzotiepinas/farmacologia , Influenza Humana/tratamento farmacológico , Influenza Humana/virologia , Vírus da Influenza A Subtipo H3N2/genética , Vírus da Influenza A Subtipo H3N2/efeitos dos fármacos , Vírus da Influenza A Subtipo H3N2/enzimologia , Triazinas/uso terapêutico , Triazinas/farmacologia , Japão , Antivirais/farmacologia , Antivirais/uso terapêutico , Morfolinas/uso terapêutico , Farmacorresistência Viral/genética , Criança , Adulto , Pré-Escolar , Adolescente , Proteínas Virais/genética , RNA Polimerase Dependente de RNA/genética , Feminino , Masculino , Tiepinas/uso terapêutico , Tiepinas/farmacologia , Lactente , Pessoa de Meia-Idade , Estações do Ano , Piridinas/uso terapêutico , Piridinas/farmacologia , Adulto Jovem , Vírus da Influenza A Subtipo H1N1/genética , Vírus da Influenza A Subtipo H1N1/efeitos dos fármacos , IdosoRESUMO
L-Leucine 5-hydroxylase (LdoA) previously found in Nostoc punctiforme PCC 73102 is a novel type of Fe(II)/α-ketoglutarate-dependent dioxygenase. LdoA catalyzed regio- and stereoselective hydroxylation of L-leucine and L-norleucine into (2S,4S)-5-hydroxyleucine and (2S)-5-hydroxynorleucine, respectively. Moreover, LdoA catalyzed sulfoxidation of L-methionine and L-ethionine in the same manner as previously described L-isoleucine 4-hydroxylase. Therefore LdoA should be a promising biocatalyst for effective production of industrially useful amino acids.
Assuntos
Dioxigenases/isolamento & purificação , Dioxigenases/metabolismo , Ferro/metabolismo , Ácidos Cetoglutáricos/metabolismo , Leucina/análogos & derivados , Leucina/metabolismo , Nostoc/enzimologia , Etionina/metabolismo , Metionina/metabolismo , Norleucina/metabolismo , Safrol/análogos & derivados , Safrol/metabolismoRESUMO
Calcium imaging has been widely adopted for its ability to record from large neuronal populations. To summarize the time course of neural activity, dimensionality reduction methods, which have been applied extensively to population spiking activity, may be particularly useful. However, it is unclear if the dimensionality reduction methods applied to spiking activity are appropriate for calcium imaging. We thus carried out a systematic study of design choices based on standard dimensionality reduction methods. We also developed a method to perform deconvolution and dimensionality reduction simultaneously (Calcium Imaging Linear Dynamical System, CILDS). CILDS most accurately recovered the single-trial, low-dimensional time courses from simulated calcium imaging data. CILDS also outperformed the other methods on calcium imaging recordings from larval zebrafish and mice. More broadly, this study represents a foundation for summarizing calcium imaging recordings of large neuronal populations using dimensionality reduction in diverse experimental settings.