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1.
Cureus ; 16(1): e52637, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-38249648

RESUMO

Background Cognition is a vital sign and its deterioration is a major concern in clinical medicine. It is usually evaluated using neuropsychological assessments, which have innate limitations such as the practice effect. To compensate for these assessments, the oscillatory power of resting-state brain activity has recently become available. The power is obtained noninvasively using magnetoencephalography and is summarized by spectral parameters such as the median frequency (MF), individual alpha frequency (IAF), spectral edge frequency 95 (SEF95), and Shannon's spectral entropy (SSE). As these parameters are less sensitive to practice effects, they are suitable for longitudinal studies. However, their reliability remains unestablished, hindering their proactive use in clinical practice. Therefore, we aimed to quantify the within-participant reliability of these parameters using repeated measurements of healthy participants to facilitate their clinical use and to evaluate the observed changes/differences in these parameters reported in previous studies. Methodology Resting-state brain activity with eyes closed was recorded using magnetoencephalography for five minutes from 15 healthy individuals (29.3 ± 4.6 years old: ranging from 23 to 28 years old). The following four spectral parameters were calculated: MF, IAF, SEF95, and SSE. To quantify reliability, the minimal detectable change (MDC) and intraclass correlation coefficient (ICC) were computed for each parameter. In addition, we used MDCs to evaluate the changes and differences in the spectral parameters reported in previous longitudinal and cross-sectional studies. Results The MDC at 95% confidence interval (MDC95) of MF, IAF, SEF95, and SSE were 0.61 Hz, 0.44 Hz, 2.91 Hz, and 0.028, respectively. The ICCs of these parameters were 0.96, 0.92, 0.94, and 0.83, respectively. The MDC95 of these parameters was smaller than the mean difference in the parameters between cognitively healthy individuals and patients with dementia, as reported in previous studies. Conclusions The spectral parameter changes/differences observed in prior studies were not attributed to measurement errors but rather reflected genuine effects. Furthermore, all spectral parameters exhibited high ICCs (>0.8), underscoring their robust within-participant reliability. Our results support the clinical use of these parameters, especially in the longitudinal monitoring and evaluation of the outcomes of interventions.

2.
EMBO J ; 40(5): e105671, 2021 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-33463726

RESUMO

The CENP-A nucleosome is a key structure for kinetochore assembly. Once the CENP-A nucleosome is established in the centromere, additional proteins recognize the CENP-A nucleosome to form a kinetochore. CENP-C and CENP-N are CENP-A binding proteins. We previously demonstrated that vertebrate CENP-C binding to the CENP-A nucleosome is regulated by CDK1-mediated CENP-C phosphorylation. However, it is still unknown how the phosphorylation of CENP-C regulates its binding to CENP-A. It is also not completely understood how and whether CENP-C and CENP-N act together on the CENP-A nucleosome. Here, using cryo-electron microscopy (cryo-EM) in combination with biochemical approaches, we reveal a stable CENP-A nucleosome-binding mode of CENP-C through unique regions. The chicken CENP-C structure bound to the CENP-A nucleosome is stabilized by an intramolecular link through the phosphorylated CENP-C residue. The stable CENP-A-CENP-C complex excludes CENP-N from the CENP-A nucleosome. These findings provide mechanistic insights into the dynamic kinetochore assembly regulated by CDK1-mediated CENP-C phosphorylation.


Assuntos
Proteína Centromérica A/metabolismo , Centrômero/metabolismo , Proteínas Cromossômicas não Histona/metabolismo , Microscopia Crioeletrônica/métodos , Cinetocoros/metabolismo , Nucleossomos/metabolismo , Animais , Centrômero/ultraestrutura , Proteína Centromérica A/ultraestrutura , Galinhas , Proteínas Cromossômicas não Histona/ultraestrutura , Cinetocoros/ultraestrutura , Modelos Moleculares , Nucleossomos/ultraestrutura , Fosforilação , Conformação Proteica
3.
Methods Mol Biol ; 2219: 49-68, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33074533

RESUMO

Here we describe methods for (a) collecting starfish during their breeding period; (b) maintaining adults with fully grown gonads in laboratory aquaria; (c) rearing fertilized eggs to brachiolaria larvae, and (d) inducing larvae to metamorphose into juveniles under laboratory conditions. Such protocols should facilitate various analyses of starfish development throughout the entire life cycle of these model organisms.


Assuntos
Asterina/crescimento & desenvolvimento , Animais , Aquicultura/instrumentação , Aquicultura/métodos , Asterina/embriologia , Desenho de Equipamento , Feminino , Larva/crescimento & desenvolvimento , Masculino , Metamorfose Biológica , Oócitos/citologia , Oogênese
4.
J Cell Biol ; 218(12): 4042-4062, 2019 12 02.
Artigo em Inglês | MEDLINE | ID: mdl-31676716

RESUMO

The kinetochore is essential for faithful chromosome segregation during mitosis. To form a functional kinetochore, constitutive centromere-associated network (CCAN) proteins are assembled on the centromere chromatin that contains the centromere-specific histone CENP-A. CENP-C, a CCAN protein, directly interacts with the CENP-A nucleosome to nucleate the kinetochore structure. As CENP-C is a hub protein for kinetochore assembly, it is critical to address how the CENP-A-CENP-C interaction is regulated during cell cycle progression. To address this question, we investigated the CENP-C C-terminal region, including a conserved CENP-A-binding motif, in both chicken and human cells and found that CDK1-mediated phosphorylation of CENP-C facilitates its binding to CENP-A in vitro and in vivo. We observed that CENP-A binding is involved in CENP-C kinetochore localization during mitosis. We also demonstrate that the CENP-A-CENP-C interaction is critical for long-term viability in human RPE-1 cells. These results provide deeper insights into protein-interaction network plasticity in centromere proteins during cell cycle progression.


Assuntos
Proteína Quinase CDC2/metabolismo , Proteína Centromérica A/metabolismo , Proteínas Cromossômicas não Histona/metabolismo , Cinetocoros/metabolismo , Animais , Centrômero/metabolismo , Galinhas , Ciclina B/metabolismo , Proteínas do Citoesqueleto/metabolismo , Feminino , Humanos , Mitose , Mutação , Oócitos/metabolismo , Fosforilação , Ligação Proteica , Proteínas Recombinantes/metabolismo , Estrelas-do-Mar
5.
Nat Cell Biol ; 20(12): 1434, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30451968

RESUMO

In the version of this Article originally published, the 'ON' and 'OFF' labels in panel c of Fig. 6 were incorrect. For the Tet treated cells (+Tet) in both image panels, CENP-T should have been 'OFF' and CENP-T Δ90 should have been 'ON'. For the cells untreated with Tet (-Tet) in both graph panels, CENP-T Δ90 should have been 'ON'. This has now been amended.

6.
Nat Cell Biol ; 20(12): 1378-1388, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30420662

RESUMO

To establish a functional kinetochore, the constitutive centromere-associated network (CCAN) forms a foundation on the centromere and recruits the KMN network, which directly binds to spindle microtubules. The CENP-C and CENP-T pathways in the CCAN recruit the KMN network to kinetochores, independently. The CENP-C pathway has been considered the major scaffold for the KMN network in vertebrate CCAN. However, we demonstrate that it is mainly the CENP-T pathway that recruits the KMN network onto the kinetochores and that CENP-T-KMN interactions are essential in chicken DT40 cells. By contrast, less Ndc80 binds to the CENP-C pathway in mitosis and the Mis12-CENP-C association is decreased during mitotic progression, which is consistent with the finding that the Mis12 complex-CENP-C binding is dispensable for cell viability. Furthermore, we find that multiple phosphoregulations of CENP-T and the Mis12 complex make the CENP-T pathway dominant. These results provide key insights into kinetochore dynamics during mitotic progression.


Assuntos
Centrômero/metabolismo , Cinetocoros/metabolismo , Microtúbulos/metabolismo , Transdução de Sinais , Fuso Acromático/metabolismo , Animais , Linhagem Celular Tumoral , Galinhas , Proteínas Cromossômicas não Histona/metabolismo , Proteínas Associadas aos Microtúbulos/metabolismo , Mitose , Complexos Multiproteicos/metabolismo , Fosforilação , Ligação Proteica
7.
PLoS One ; 11(12): e0168588, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27992543

RESUMO

PURPOSE: Quantitative imaging of neuromagnetic fields based on automated region of interest (ROI) setting was analyzed to determine the characteristics of cerebral neural activity in ischemic areas. METHODS: Magnetoencephalography (MEG) was used to evaluate spontaneous neuromagnetic fields in the ischemic areas of 37 patients with unilateral internal carotid artery (ICA) occlusive disease. Voxel-based time-averaged intensity of slow waves was obtained in two frequency bands (0.3-4 Hz and 4-8 Hz) using standardized low-resolution brain electromagnetic tomography (sLORETA) modified for a quantifiable method (sLORETA-qm). ROIs were automatically applied to the anterior cerebral artery (ACA), anterior middle cerebral artery (MCAa), posterior middle cerebral artery (MCAp), and posterior cerebral artery (PCA) using statistical parametric mapping (SPM). Positron emission tomography with 15O-gas inhalation (15O-PET) was also performed to evaluate cerebral blood flow (CBF) and oxygen extraction fraction (OEF). Statistical analyses were performed using laterality index of MEG and 15O-PET in each ROI with respect to distribution and intensity. RESULTS: MEG revealed statistically significant laterality in affected MCA regions, including 4-8 Hz waves in MCAa, and 0.3-4 Hz and 4-8 Hz waves in MCAp (95% confidence interval: 0.020-0.190, 0.030-0.207, and 0.034-0.213), respectively. We found that 0.3-4 Hz waves in MCAp were highly correlated with CBF in MCAa and MCAp (r = 0.74, r = 0.68, respectively), whereas 4-8 Hz waves were moderately correlated with CBF in both the MCAa and MCAp (r = 0.60, r = 0.63, respectively). We also found that 4-8 Hz waves in MCAp were statistically significant for misery perfusion identified on 15O-PET (p<0.05). CONCLUSIONS: Quantitatively imaged spontaneous neuromagnetic fields using the automated ROI setting enabled clear depiction of cerebral ischemic areas. Frequency analysis may reveal unique neural activity that is distributed in the impaired vascular metabolic territory, in which the cerebral infarction has not yet been completed.


Assuntos
Isquemia Encefálica/diagnóstico , Doenças das Artérias Carótidas/diagnóstico , Artéria Carótida Interna/diagnóstico por imagem , Magnetoencefalografia/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Mapeamento Encefálico , Circulação Cerebrovascular , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Tomografia por Emissão de Pósitrons
8.
Open Neuroimag J ; 10: 85-101, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27708745

RESUMO

We studied sex-related differences in gamma oscillation during an auditory oddball task, using magnetoencephalography and electroencephalography assessment of imaginary coherence (IC). We obtained a statistical source map of event-related desynchronization (ERD) / event-related synchronization (ERS), and compared females and males regarding ERD / ERS. Based on the results, we chose respectively seed regions for IC determinations in low (30-50 Hz), mid (50-100 Hz) and high gamma (100-150 Hz) bands. In males, ERD was increased in the left posterior cingulate cortex (CGp) at 500 ms in the low gamma band, and in the right caudal anterior cingulate cortex (cACC) at 125 ms in the mid-gamma band. ERS was increased in the left rostral anterior cingulate cortex (rACC) at 375 ms in the high gamma band. We chose the CGp, cACC and rACC as seeds, and examined IC between the seed and certain target regions using the IC map. IC changes depended on the height of the gamma frequency and the time window in the gamma band. Although IC in the mid and high gamma bands did not show sex-specific differences, IC at 30-50 Hz in males was increased between the left rACC and the frontal, orbitofrontal, inferior temporal and fusiform target regions. Increased IC in males suggested that males may acomplish the task constructively, analysingly, emotionally, and by perfoming analysis, and that information processing was more complicated in the cortico-cortical circuit. On the other hand, females showed few differences in IC. Females planned the task with general attention and economical well-balanced processing, which was explained by the higher overall functional cortical connectivity. CGp, cACC and rACC were involved in sex differences in information processing and were likely related to differences in neuroanatomy, hormones and neurotransmitter systems.

9.
J Cell Sci ; 129(16): 3153-66, 2016 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-27390173

RESUMO

Extracellular ligands control biological phenomena. Cells distinguish physiological stimuli from weak noise stimuli by establishing a ligand-concentration threshold. Hormonal control of the meiotic G2/M transition in oocytes is essential for reproduction. However, the mechanism for threshold establishment is unclear. In starfish oocytes, maturation-inducing hormones activate the PI3K-Akt pathway through the Gßγ complex of heterotrimeric G-proteins. Akt directly phosphorylates both Cdc25 phosphatase and Myt1 kinase, resulting in activation of cyclin-B-Cdk1, which then induces meiotic G2/M transition. Here, we show that cyclin-B-Cdk1 is partially activated after subthreshold hormonal stimuli, but this triggers negative feedback, resulting in dephosphorylation of Akt sites on Cdc25 and Myt1, thereby canceling the signal. We also identified phosphatase activity towards Akt substrates that exists independent of stimuli. In contrast to these negative regulatory activities, an atypical Gßγ-dependent pathway enhances PI3K-Akt-dependent phosphorylation. Based on these findings, we propose a model for threshold establishment in which hormonal dose-dependent competition between these new pathways establishes a threshold; the atypical Gßγ-pathway becomes predominant over Cdk-dependent negative feedback when the stimulus exceeds this threshold. Our findings provide a regulatory connection between cell cycle and signal transduction machineries.


Assuntos
Proteína Quinase CDC2/metabolismo , Ciclina B/metabolismo , Fase G2 , Meiose , Mitose , Estrelas-do-Mar/citologia , Estrelas-do-Mar/metabolismo , Adenina/análogos & derivados , Adenina/farmacologia , Animais , Ativação Enzimática/efeitos dos fármacos , Retroalimentação Fisiológica/efeitos dos fármacos , Fase G2/efeitos dos fármacos , Subunidades beta da Proteína de Ligação ao GTP/metabolismo , Humanos , Meiose/efeitos dos fármacos , Mitose/efeitos dos fármacos , Modelos Biológicos , Proteínas Mutantes/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação/efeitos dos fármacos , Fosfosserina/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Estrelas-do-Mar/efeitos dos fármacos , Especificidade por Substrato/efeitos dos fármacos , Fosfatases cdc25/metabolismo
10.
Brain Behav ; 6(3): e00427, 2016 03.
Artigo em Inglês | MEDLINE | ID: mdl-26855826

RESUMO

INTRODUCTION: Brain-imaging techniques have begun to be popular in evaluating the effectiveness of cognitive intervention training. Although gamma activities are rarely used as an index of training effects, they have several characteristics that suggest their potential suitability for this purpose. This pilot study examined whether cognitive training in elderly people affected the high-gamma activity associated with attentional processing and whether high-gamma power changes were related to changes in behavioral performance. METHODS: We analyzed (MEG) magnetoencephalography data obtained from 35 healthy elderly subjects (60-75 years old) who had participated in our previous intervention study in which the subjects were randomly assigned to one of the three types of intervention groups: Group V trained in a vehicle with a newly developed onboard cognitive training program, Group P trained with a similar program but on a personal computer, and Group C was trained to solve a crossword puzzle as an active control group. High-gamma (52-100 Hz) activity during a three-stimulus visual oddball task was measured before and after training. As a result of exclusion in the MEG data analysis stage, the final sample consisted of five subjects in Group V, nine subjects in Group P, and seven subjects in Group C. RESULTS: Results showed that high-gamma activities were differently altered between groups after cognitive intervention. In particular, members of Group V, who showed significant improvements in cognitive function after training, exhibited increased high-gamma power in the left middle frontal gyrus during top-down anticipatory target processing. High-gamma power changes in this region were also associated with changes in behavioral performance. CONCLUSIONS: Our preliminary results suggest the usefulness of high-gamma activities as an index of the effectiveness of cognitive training in elderly subjects.


Assuntos
Encéfalo/fisiologia , Terapia Cognitivo-Comportamental/métodos , Córtex Pré-Frontal/fisiologia , Idoso , Atenção/fisiologia , Cognição/fisiologia , Feminino , Lobo Frontal , Humanos , Magnetoencefalografia , Masculino , Pessoa de Meia-Idade , Neuroimagem/métodos , Projetos Piloto
11.
Clin Neurophysiol ; 127(1): 179-186, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26073183

RESUMO

OBJECTIVE: We hypothesized that high frequency oscillations (HFOs) are differently suppressed during rapid eye movement sleep (REM) between epileptogenic and less epileptogenic cortices, and that the suppressive effect can serve as a specific marker of epileptogenicity. METHODS: Intracranial electroencephalography (EEG) was recorded in 13 patients with drug-resistant epilepsy. HFOs between 80 and 200Hz were semi-automatically detected from total 15-min EEG epochs each for REM and slow wave sleep (SWS). z-Score of HFO occurrence rate was calculated from the baseline rate derived from non-epileptogenic cortex. Intracranial electrodes were labeled as REM dominant HFO (RdH) if REM z-score was greater than SWS z-score or as SWS dominant HFO (SdH) if SWS z-score was greater than REM z-score. Relationship of electrode location to the area of surgical resection was compared between RdH and SdH electrodes. RESULTS: Out of 1070 electrodes, 101 were defined as RdH electrodes and 115 as SdH electrodes. RdH electrodes were associated with the area of resection in patients with postoperative seizure freedom (P<0.001), but not in patients without seizure freedom. CONCLUSIONS: HFOs near the epileptogenic zone are less suppressed during REM. SIGNIFICANCE: The less suppressive effect of REM may provide a specific marker of epileptogenicity.


Assuntos
Potenciais de Ação , Epilepsia Resistente a Medicamentos/fisiopatologia , Sono REM , Adolescente , Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
12.
PLoS One ; 10(2): e0116918, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25659131

RESUMO

Deficits in attentional-inhibitory control have been reported to correlate to anger, hostility, and aggressive behavior; therefore, inhibitory control appears to play an important role in prosocial behavior. Moreover, recent studies have demonstrated that oxytocin (OT) exerts a prosocial effect (e.g., decreasing negative behaviors, such as aggression) on humans. However, it is unknown whether the positively valenced effect of OT on sociality is associated with enhanced attentional-inhibitory control. In the present study, we hypothesized that OT enhances attentional-inhibitory control and that the positively valenced effect of OT on social cognition is associated with enhanced attentional-inhibitory control. In a single-blind, placebo-controlled crossover trial, we tested this hypothesis using 20 healthy male volunteers. We considered a decrease in the hostility detection ratio, which reflects the positively valenced interpretation of other individuals' facial expressions, to be an index of the positively valenced effects of OT (we reused the results of our previously published study). As a measure of attentional-inhibitory control, we employed a modified version of the flanker task (i.e., a shorter conflict duration indicated higher inhibitory control). These results failed to demonstrate any significant behavioral effects of OT (i.e., neither a positively valenced effect on facial cognition nor an effect on attentional-inhibitory control). However, the enhancement of attentional-inhibitory control after OT administration significantly correlated to the positively valenced effects on the interpretation of uncertain facial cognition (i.e., neutral and ambiguous facial expressions).


Assuntos
Atenção/efeitos dos fármacos , Emoções Manifestas , Ocitocina/administração & dosagem , Percepção Visual/efeitos dos fármacos , Adulto , Face , Humanos , Masculino , Pessoa de Meia-Idade
13.
Neuroimage ; 100: 290-300, 2014 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-24960420

RESUMO

The current study used a magnetoencephalogram to investigate the relationship between high-gamma (52-100 Hz) activity within an attention network and individual differences in behavioral performance among healthy elderly adults. We analyzed brain activity in 41 elderly subjects performing a 3-stimulus visual oddball task. In addition to the average amplitude of event-related fields in the left intraparietal sulcus (IPS), high-gamma power in the left middle frontal gyrus (MFG), the strength of high-gamma imaginary coherence between the right MFG and the left MFG, and those between the right MFG and the left thalamus predicted individual differences in reaction time. In addition, high-gamma power in the left MFG was correlated with task accuracy, whereas high-gamma power in the left thalamus and left IPS was correlated with individual processing speed. The direction of correlations indicated that higher high-gamma power or coherence in an attention network was associated with better task performance and, presumably, higher cognitive function. Thus, high-gamma activity in different regions of this attention network differentially contributed to attentional processing, and such activity could be a fundamental process associated with individual differences in cognitive aging.


Assuntos
Atenção/fisiologia , Lobo Frontal/fisiologia , Ritmo Gama/fisiologia , Individualidade , Magnetoencefalografia/métodos , Rede Nervosa/fisiologia , Desempenho Psicomotor/fisiologia , Tálamo/fisiologia , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
14.
Clin Neurophysiol ; 125(12): 2358-63, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24856458

RESUMO

OBJECTIVE: Small focal cortical dysplasia (FCD) may be ambiguous or overlooked on magnetic resonance (MR) imaging. Source localization of EEG and magnetoencephalography (MEG) spikes was evaluated to confirm the diagnosis of small FCD. METHODS: This study included 6 epilepsy patients with a single small lesion on MR imaging suggesting FCD within a single gyrus among 181 consecutive epilepsy patients admitted to our epilepsy monitoring unit over 27 months. Stereotypical interictal spikes were detected on simultaneous EEG and MEG recordings and the onset-related source of averaged spikes was estimated. RESULTS: All 6 patients had unique clinical characteristics as follows: leg sensori-motor seizures in 5 patients and eye version in 1 patient; a small MR imaging lesion suggesting FCD in the dorsal peri-rolandic region, which had been overlooked until our evaluation; and both EEG and MEG dipoles were estimated adjacent to the MR imaging lesion. CONCLUSIONS: Source localization of EEG and MEG spikes can confirm the diagnosis of FCD based on a single small MR imaging lesion, which was overlooked by previous examination of MR images. SIGNIFICANCE: Examination of MR images should be based on spike source localization as well as seizure semiology to identify subtle MR imaging abnormalities.


Assuntos
Potenciais de Ação , Eletroencefalografia , Epilepsia/fisiopatologia , Magnetoencefalografia , Malformações do Desenvolvimento Cortical/fisiopatologia , Córtex Motor/fisiopatologia , Adolescente , Adulto , Eletroencefalografia/métodos , Epilepsia/diagnóstico , Feminino , Humanos , Magnetoencefalografia/métodos , Masculino , Malformações do Desenvolvimento Cortical/diagnóstico , Córtex Motor/anormalidades , Adulto Jovem
15.
J Cell Biol ; 204(6): 881-9, 2014 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-24616226

RESUMO

Entry into M phase is governed by cyclin B-Cdk1, which undergoes both an initial activation and subsequent autoregulatory activation. A key part of the autoregulatory activation is the cyclin B-Cdk1-dependent inhibition of the protein phosphatase 2A (PP2A)-B55, which antagonizes cyclin B-Cdk1. Greatwall kinase (Gwl) is believed to be essential for the autoregulatory activation because Gwl is activated downstream of cyclin B-Cdk1 to phosphorylate and activate α-endosulfine (Ensa)/Arpp19, an inhibitor of PP2A-B55. However, cyclin B-Cdk1 becomes fully activated in some conditions lacking Gwl, yet how this is accomplished remains unclear. We show here that cyclin B-Cdk1 can directly phosphorylate Arpp19 on a different conserved site, resulting in inhibition of PP2A-B55. Importantly, this novel bypass is sufficient for cyclin B-Cdk1 autoregulatory activation. Gwl-dependent phosphorylation of Arpp19 is nonetheless necessary for downstream mitotic progression because chromosomes fail to segregate properly in the absence of Gwl. Such a biphasic regulation of Arpp19 results in different levels of PP2A-B55 inhibition and hence might govern its different cellular roles.


Assuntos
Asterina/enzimologia , Proteína Quinase CDC2/metabolismo , Ciclina B/metabolismo , Proteína Fosfatase 2/metabolismo , Animais , Asterina/citologia , Células Cultivadas , Segregação de Cromossomos , Ativação Enzimática , Meiose , Fosfoproteínas/metabolismo , Fosforilação , Processamento de Proteína Pós-Traducional , Coelhos
16.
Methods Mol Biol ; 1128: 311-30, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24567224

RESUMO

Antibodies are widely utilized in cell and molecule biology for immunoblots, immunostaining, immunoprecipitation, immunoaffinity purification, and immunoassay. Some antibodies can be used for in vivo inhibition experiments. These antibodies bind to their target molecules and neutralize their functions, providing functional information in the study of their biological role. Here, we describe our methods for obtaining inhibitory antibodies against desired proteins. We then describe in the starfish oocyte system how to inhibit a target protein, even in the nucleus, by injection of antibody into the cytoplasm, and how to evaluate antibody inhibition of cell cycle regulators in small numbers of oocytes.


Assuntos
Anticorpos/farmacologia , Oócitos/efeitos dos fármacos , Estrelas-do-Mar/efeitos dos fármacos , Animais , Anticorpos/isolamento & purificação , Técnicas de Cultura de Células , Separação Celular , Sistema Livre de Células , Células Cultivadas , Ensaios Enzimáticos , Microinjeções , Oócitos/enzimologia , Protamina Quinase/antagonistas & inibidores , Protamina Quinase/imunologia , Protamina Quinase/metabolismo , Coelhos , Estrelas-do-Mar/citologia
17.
PLoS One ; 8(11): e80126, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24278247

RESUMO

Optimal brain sensitivity to the fundamental frequency (F0) contour changes in the human voice is important for understanding a speaker's intonation, and consequently, the speaker's attitude. However, whether sensitivity in the brain's response to a human voice F0 contour change varies with an interaction between an individual's traits (i.e., autistic traits) and a human voice element (i.e., presence or absence of communicative action such as calling) has not been investigated. In the present study, we investigated the neural processes involved in the perception of F0 contour changes in the Japanese monosyllables "ne" and "nu." "Ne" is an interjection that means "hi" or "hey" in English; pronunciation of "ne" with a high falling F0 contour is used when the speaker wants to attract a listener's attention (i.e., social intonation). Meanwhile, the Japanese concrete noun "nu" has no communicative meaning. We applied an adaptive spatial filtering method to the neuromagnetic time course recorded by whole-head magnetoencephalography (MEG) and estimated the spatiotemporal frequency dynamics of event-related cerebral oscillatory changes in beta band during the oddball paradigm. During the perception of the F0 contour change when "ne" was presented, there was event-related de-synchronization (ERD) in the right temporal lobe. In contrast, during the perception of the F0 contour change when "nu" was presented, ERD occurred in the left temporal lobe and in the bilateral occipital lobes. ERD that occurred during the social stimulus "ne" in the right hemisphere was significantly correlated with a greater number of autistic traits measured according to the Autism Spectrum Quotient (AQ), suggesting that the differences in human voice processing are associated with higher autistic traits, even in non-clinical subjects.


Assuntos
Transtorno Autístico/fisiopatologia , Encéfalo/fisiopatologia , Voz , Adulto , Feminino , Humanos , Magnetoencefalografia , Masculino , Adulto Jovem
18.
PLoS One ; 8(9): e73416, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24023873

RESUMO

Unilateral spatial neglect (USN) is one of the most common symptoms of right hemisphere damage; its classical symptom is that patients fail to respond to information on their left side. It has been postulated that disturbance of 2 separate attentional networks relates to the occurrence of USN. However, little is known about the underlying mechanism and neuronal substrates. In this study, we measured spontaneous neural activity by means of magnetoencephalography in 13 patients with brain damage and 5 control subjects. To study the relationship between functional connectivity at rest and severity of USN symptoms, we determined the imaginary coherence values relating to the inter-hemispherical ventral and dorsal attentional networks, as well as the clinical severity of USN using neuropsychological tests and behavioral rating scales. The present results showed that inter-hemispherical connectivity in the ventral attentional network, especially between the left and right angular gyri, detected in the alpha band is associated with the severity of USN symptoms. This may suggest that connectivity of inter-hemispherical homologous regions of the ventral attentional network in the alpha band could be one of the biomarkers of attentional network imbalance occurring in patients with USN.


Assuntos
Ritmo alfa/fisiologia , Atenção/fisiologia , Cérebro/fisiopatologia , Rede Nervosa/fisiopatologia , Transtornos da Percepção/fisiopatologia , Descanso/fisiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Magnetoencefalografia , Masculino , Pessoa de Meia-Idade , Transtornos da Percepção/diagnóstico
19.
Clin Neurophysiol ; 124(11): 2091-100, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-23756060

RESUMO

OBJECTIVE: Corpus callosotomy limits the bilateral synchrony of epileptic discharges. However, the instantaneous changes in bilateral synchrony during corpus callosotomy are unclear. The present study investigated how and when bilateral synchrony is suppressed in the anterior and then posterior steps of corpus callosotomy. METHODS: Intra-operative scalp electroencephalography (EEG) was recorded simultaneously with surgical video for six patients who underwent total corpus callosotomy for medically intractable drop attacks. The time-varying EEG inter-hemispheric coherence was quantified by wavelet transform coherence and trend analysis. RESULTS: The 4-13 Hz coherence decreased after corpus callosotomy in five patients. Significant decrease in coherence was observed only during the posterior step of callosal sectioning in three patients, but throughout both steps in two patients. CONCLUSIONS: Decrease in inter-hemispheric coherence is not always correlated with the stages of callosal sectioning. Inter-hemispheric coherence is decreased during the final stage of corpus callosotomy and the effect is maximized after sectioning is completed. SIGNIFICANCE: Various patterns of coherence decrease suggest individual variations in the participation of the corpus callosum in the genesis of bilateral synchrony. Time-varying inter-hemispheric EEG coherence is useful to monitor the physiological completeness of corpus callosotomy.


Assuntos
Corpo Caloso/fisiopatologia , Corpo Caloso/cirurgia , Eletroencefalografia , Epilepsia/fisiopatologia , Epilepsia/cirurgia , Lateralidade Funcional , Monitorização Intraoperatória , Criança , Pré-Escolar , Feminino , Seguimentos , Humanos , Masculino , Modelos Neurológicos , Cuidados Paliativos , Fatores de Tempo , Análise de Ondaletas
20.
Open Neuroimag J ; 7: 15-26, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23750187

RESUMO

BACKGROUND: We studied the imaginary coherence (IC) of gamma frequency oscillations between brain regions of male schizophrenia patients during an auditory oddball task using magnetoencephalography (MEG) and electroencephalography (EEG). METHODS: Subjects were 10 right-handed male schizophrenia patients, evaluated by the positive and negative symptom scale (PANSS), and 10 healthy controls. Functional connectivity during the auditory oddball task was reconstructed in low (30-50 Hz) and high (50-100 Hz) gamma bands, and represented by imaginary coherence (IC) based on significant oscillatory power changes. We calculated correlations between PANSS scores and IC. RESULTS: In the high gamma band, IC between left occipital and right prefrontal lobe areas during the time window 750-1000 ms from stimulus onset showed negative correlations with total negative scores, total positive scores, the sum of positive and negative scores in PANSS, conceptual disorganization, and social avoidance scores. In the low gamma band, IC between the same areas from 250-500 ms also showed a negative correlation with the conceptual disorganization score. In the same time window, IC between left occipital and right frontoparietal lobe areas in the low gamma band showed a positive correlation with hallucinatory behavior; IC between right temporal pole and left prefrontal lobe areas showed a positive correlation with delusion scores, although these ICs were decreased relative to controls. CONCLUSIONS: Functional disconnection of high and low gamma bands in auditory oddball task may play an important role in the auditory processing in schizophrenia patients.

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