RNA-Activated Protein Kinase, PKR

The double-stranded RNA (dsRNA) activated protein kinase, PKR, is one of the several enzymes induced by interferons and a key molecule mediating the antiviral effects of interferons. PKR contain an N-terminal, double-stranded RNA binding domain (dsRBD), which has two tandem copies of the motifs (dsRBM I and dsRBM II). Upon binding to viral dsRNA, PKR is activated via autophosphorylation. Activated PKR has several substrates; one of the examples is eukaryotic translation initiation factor 2 (eIF2a). The phosphorylation of eIF2a leads to the termination of cell growth by inhibiting protein synthesis in response to viral infection. The objective of this project was to characterize the dsRBM I and define the dsRNA binding using biophysical methods. First, the dsRBM I gene was cloned from a pET-28b to a pET-11a expression plasmid. N-terminal poly-histidine tags on pET-28b are for affinity purification; however, these tags can alter the structure and function of proteins, thus the gene of dsRBM I was transferred into the plasmid without tags (pET-11a) and expressed as a native protein. The dsRBM I was transformed into and expressed by Rosetta DE3plyS expression cells. Purification was done by FPLC using a Sepharose IEX ion exchange followed by Heparin affinity column; yielding pure protein was assayed by PAGE. Analytical Ultracentrifugation, Sedimentation Velocity, was used to characterize free solution association state and hydrodynamic properties of the protein. The slight decrease in S-value with concentration is due to the hydrodynamic non-ideality. No self association was observed. The obtained molecule weight was 10,079 Da. The calculated sedimentation constant at zero concentration at 20°C in water was 1.23 and its friction coefficient was 3.575 ´ 10-8. The frictional ratio of sphere and dsRBM I became 1.30. Therefore, dsRBM I must be non-globular and more asymmetric shape. Isolated dsRBM I exhibits the same tertiary fold as compared to context in the full domain but it exhibited weaker binding affinity than full domain to a 20 bp dsRNA. However, when the conditions allowed for its saturation, dsRBM I to 20 bp dsRNA has similar stoichiometry as full dsRBD

The mineral assemblage of symplectites in lunar meteorite Asuka-881757

Asuka-881757 with a gabbro-like texture is from lunar mare. It is different from other known lunar mare basaltic meteorites. It is coarse-grained basalt with symplectite texture in mesostasis. It is composed mainly of pyroxene and maskelynite (An_∿An_). Most ilmenites, troitites and some Fe-Ni metals are surrounded by symplectite. One of the symplectites consists of very fine-grained Fe-rich olivine (Fa_) and silica phase in pyroxene host around troilite. However, the troilite and primary pyroxene did not react to form the symplectite. The coexisting fayalite, hedenbergitic pyroxene and silica phase suggest that they crystallized from the primary melt at a pressure lower than 1.15GPa, indicating formation near the lunar surface

Behavioral destabilization induced by the selective serotonin reuptake inhibitor fluoxetine

<p>Abstract</p> <p>Background</p> <p>Selective serotonin reuptake inhibitors (SSRIs) are widely used to treat mood and anxiety disorders. However, neuronal bases for both beneficial and adverse effects of SSRIs remain poorly understood. We have recently shown that the SSRI fluoxetine can reverse the state of maturation of hippocampal granule cells in adult mice. The granule cell "dematuration" is induced in a large population of granule cells, and greatly changes functional and physiological properties of these cells. Here we show that this unique form of neuronal plasticity is correlated with a distinct change in behavior of mice.</p> <p>Results</p> <p>We chronically treated adult male mice with fluoxetine, and examined its effect on several forms of behavior of mice. During fluoxetine treatments, mice showed a marked increase in day-to-day fluctuations of home cage activity levels that was characterized by occasional switching between hypoactivity and hyperactivity within a few days. This destabilized cage activity was accompanied by increased anxiety-related behaviors and could be observed up to 4 weeks after withdrawal from fluoxetine. As reported previously, the granule cell dematuration by fluoxetine includes a reduction of synaptic facilitation at the granule cell output, mossy fiber, synapse to the juvenile level. Mossy fiber synaptic facilitation examined electrophysiologically in acute hippocampal slices also remained suppressed after fluoxetine withdrawal and significantly correlated with the fluctuation of cage activity levels in individual mice. Furthermore, in mice lacking the 5-HT₄receptor, in which the granule cell dematuration has been shown to be attenuated, fluoxetine had no significant effect on the fluctuation of cage activity levels.</p> <p>Conclusions</p> <p>Our results demonstrate that the SSRI fluoxetine can induce marked day-to-day changes in activity levels of mice in the familiar environment, and that the dematuration of the hippocampal granule cells is closely associated with the expression of this destabilized behavior. Based on these results, we propose that the granule cell dematuration can be a potential cellular basis underlying switching-like changes in the behavioral state associated with SSRI treatments.</p

Spherical functions of hermitian and symmetric forms

Thesis--University of Tsukuba, D.Sc.(B), no. 450, 1988. 3. 2

Melanocortin Systems on Pigment Dispersion in Fish Chromatophores

α-Melanocyte-stimulating hormone (α-MSH) is responsible for pigment dispersion in the chromatophores of fish and other tetrapods such as amphibians and reptiles. Recently, we discovered that α-MSH did not always stimulate pigment dispersion because this hormonal peptide exerted no effects on the melanophores of flounders. We assumed that the reduction of α-MSH activity was related to the co-expression of different α-MSH receptor subtypes – termed melanocortin receptors (MCR) – a member of G-protein-coupled receptors (GPCR) – based on several reports demonstrating that GPCR forms heterodimers with various properties that are distinct from those of the corresponding monomers. In this review, we summarize the relationships between the pigment-dispersing activity of α-MSH-related peptides, molecular forms of α-MSH-related peptides, and mcr subtypes expressed in fish chromatophores

A novel behavioral science-based health checkup program and subsequent metabolic risk reductions in a workplace: Checkup championship

The effectiveness of general health checkups and lifestyle counseling has been questioned. This study examined whether a workplace health promotion program implemented during a health checkup was associated with metabolic syndrome-related indicators. Hakuhodo DY group, one of Japan's largest advertising agencies, implemented a behavioral science-based program called "Checkup Championship" (Kenshin-sen in Japanese) in 2019, in which all employees could voluntarily participate. We studied 3697 employees (2818 men and 879 women, mean age: 40.7 years), consisting of 1509 program participants and 2188 non-participants. The characteristics of participants and non-participants were balanced using inverse probability weighting. We used their data from the health checkups in 2018 and 2019 together with other covariates and performed a difference-in-differences analysis using a linear mixed model. After program implementation, greater reductions were observed among participants compared with non-participants in weight (-0.66 kg, 95% confidence interval: -0.84 to -0.47), body mass index (-0.23 kg/m², -0.29 to -0.16), waist circumference (-0.67 cm, -0.91 to -0.43), systolic blood pressure (-1.13 mmHg, -2.10 to -0.16), and diastolic blood pressure (-0.84 mmHg, -1.53 to -0.15). In addition, we observed greater reductions in weight, body mass index, waist circumference, and low-density lipoprotein cholesterol among participants who were with two or more risk factors for metabolic syndrome than other participants. We found that participation in a health checkup program based on behavioral science was associated with reduced metabolic syndrome-related indicators. There may be room for improvement in the effectiveness of general health checkups

Validation of the Japanese version of the CPAQ-8

This study aimed to examine the reliability and validity of the Japanese version of the Chronic Pain Acceptance Questionnaire (CPAQ-8J). A total of 108 outpatients with chronic pain completed the CPAQ-8 questionnaire, along with the Acceptance and Action Questionnaire-II, Hospital Anxiety and Depression Scale, Pain Disability Assessment Scale, Numerical Rating Scale, and EuroQol 5 dimensions 5-level. Confirmatory factor analyses examined the factor structure. Results indicated that the CPAQ-8J comprised a two-component factor structure. Correlations between the CPAQ-8J and each variable were as expected, except between the “pain willingness” subscale and other scales ; thus, the CPAQ-8J had a certain degree of convergent validity. Internal consistency and test-retest reliability suggest that the CPAQ-8J is reliable. The psychometric properties of the CPAQ-8J meet a certain standard ; meanwhile, some issues must be addressed for its practical application. Further research should consider the influence of cultural characteristics in practical application

Efficacy of topiramate for intractable childhood generalized epilepsy with epileptic spasms: With special reference to electroencephalographic changes

AbstractPurposeEpileptic spasms (ES) beyond infancy are a highly refractory type of seizures that require the development of an effective treatment. We therefore studied the efficacy and safety of topiramate (TPM), which is a drug that is indicated to be effective for intractable childhood epilepsy, for ES.MethodsOut of 58 children with ES, we enrolled 33 patients treated with TPM at ≤12years of age. The administration of TPM was limited to cases of epilepsies that were resistant to any other potent treatment. We retrospectively investigated the efficacy of TPM for seizures and changes in electroencephalogram (EEG) findings.ResultsThe median age at the start of TPM treatment was 5years, 8months. All patients had ES and 28 also had tonic seizures. As for the efficacy of TPM for all seizures, five patients became seizure-free and two had a ≥50% reduction in seizures. Seizure aggravation was observed in six patients. Of 29 patients whose EEG findings were compared before and during TPM treatment, nine showed EEG improvement with reduced epileptic discharges. Adverse effects were observed in 13 patients and included somnolence, anorexia, and irritability. In general, TPM was well tolerated.ConclusionsTPM can be effective at suppressing very intractable ES in a proportion of patients who do not respond to any other treatment. The efficacy of TPM may be predictable based on EEG changes observed early in the course of treatment. TPM is promising for the treatment of extremely intractable childhood epilepsy and it has largely tolerable adverse effects