Structure-Substrate Binding Relationships of HIV-1 Reverse Transcriptase

Human Immunodeficiency Virus, type 1 (HIV-1), is the causative agent of the Acquired Immunodeficiency Syndrome (AIDS). HIV-1 reverse transcriptase (RT), a heterodimer p66/p51, has been the major target for treatment of AIDS. The significance of the p51 subunit and the RNase H domain of p66 in terms of their influence on the RNA-dependent DNA synthesis was investigated. Clones of the wildtype HIV-1 RT subunits, p66 and p51, and a recombinant C-terminal deletion mutant, p64, [Barr, P. J. (1987) Bio/Technoloav 5, 486-489] were employed to study the structure-substrate binding relationships of HIV-1 RT. The activity assays of RNA-dependent DNA synthesis on both poly(rA)(dT) and a random base RNA template hybridized with a DNA oligomer showed that p51 significantly affects the enzyme activity. The increase in processivity by p51 in the p66/p51 heterodimer was also demonstrated. These observations suggested that the integrity of p51 is important in subunit-interactions for maintaining a favorable conformation of the enzyme for optimal function. C-terminal deletion in p66 was seen to decrease the processivity. The dissociation constant (Kd) for poly(rA)(dT) obtained by nitrocellulose binding assays suggested that the processivity of HIV 1 RT on poly(rA)(dT) correlated with the affinity for the substrate. The processivity of RT on RNA335-DNA20 was seen to be affected by the pause sites observed on the autoradiograms. The pauses of DNA synthesis tended to occur at positions of template containing poly G-C sequences. The order of processivity observed on RNA335-DNA20 was p64/p64, p66/p66 \u3c p64/p51 \u3c p66/p51. The C-terminal deletion in p66 was shown to affect the ability to extend the DNA strand on RNA template. In those non-wildtype forms of HIV-1 RT (p66/p66, p64/p64, and p64/p51), the affinity for primer-template seemed to be sensitive to the structure of the RNA template as seen when comparing Kds between poly(rA)(dT) and RNA335-DNA20. The wildtype enzyme, p66/p51, appeared to have a similar affinity for both substrates

多様な拘束圧及びせん断 ひずみ条件における砂の粒子破砕に関する研究

首都大学東京, 2015-09-30, 博士（理学）, 甲第583号首都大学東

Comorbidity and confounding factors in attention-deficit/hyperactivity disorder and sleep disorders in children

Sleep problems are commonly reported in children with attention-deficit/hyperactivity disorder (ADHD) symptoms. Research data regarding the complex and reciprocal relationship between ADHD and sleep disturbances has now accumulated. This paper is focused on the types of sleep problems that are associated with ADHD symptomatology, and attempts to untangle confounding factors and overlapping symptoms. The goal is also to present an updated overview of the pathophysiology of and treatment strategies for sleep problems in children with ADHD. The review also points out that future research will be needed to clarify further the other psychiatric comorbidities and side effects of medication in order to improve treatment outcomes and prevent misdiagnosis in clinical practice

Anserine Reverses Exercise-Induced Oxidative Stress and Preserves Cellular Homeostasis in Healthy Men

The study tested whether anserine (beta-alanyl-3-methyl-l-histidine), the active ingredient of chicken essence affects exercise-induced oxidative stress, cell integrity, and haematology biomarkers. In a randomized placebo-controlled repeated-measures design, ten healthy men ingested anserine in either a low dose (ANS-LD) 15 mg·kg−1·bw−1, high dose (ANS-HD) 30 mg·kg−1·bw−1, or placebo (PLA), following an exercise challenge (time to exhaustion), on three separate occasions. Anserine supplementation increased superoxide dismutase (SOD) by 50% (p < 0.001, effect size d = 0.8 for both ANS-LD and ANS-HD), and preserved catalase (CAT) activity suggesting an improved antioxidant activity. However, both ANS-LD and ANS-HD elevated glutathione disulfide (GSSG), (both p < 0.001, main treatment effect), and consequently lowered the glutathione to glutathione disulfide (GSH/GSSG) ratio compared with PLA (p < 0.01, main treatment effect), without significant effects on thiobarbituric acid active reactive substances (TBARS). Exercise-induced cell damage biomarkers of glutamic-oxaloacetic transaminase (GOT) and myoglobin were unaffected by anserine. There were slight but significant elevations in glutamate pyruvate transaminase (GPT) and creatine kinase isoenzyme (CKMB), especially in ANS-HD (p < 0.05) compared with ANS-LD or PLA. Haematological biomarkers were largely unaffected by anserine, its dose, and without interaction with post exercise time-course. However, compared with ANS-LD and PLA, ANS-HD increased the mean cell volume (MCV), and decreased the mean corpuscular haemoglobin concentration (MCHC) (p < 0.001). Anserine preserves cellular homoeostasis through enhanced antioxidant activity and protects cell integrity in healthy men, which is important for chronic disease prevention. However, anserine temporal elevated exercise-induced cell-damage, together with enhanced antioxidant activity and haematological responses suggest an augmented exercise-induced adaptative response and recovery

BIOMECHANICAL ANALYSIS OF BADMINTON DIFFERENT FORWARD STEPS

The purpose of this study was to compare the biomechanical variables between 2-step and 3-step forward steps in badminton. Eight collegiate elite male badminton players participated in this study. Eight Vicon T-20 cameras (300 Hz) were used to record the 3D kinematics data and a Kistler force plate (1500 Hz) was used to collect the GRF data of the last steps. A Wilcoxon matched-pairs signed-rank nonparametric statistical test was conducted to compare the differences between two kinds of forward step movements. The results showed that the movement time for 3-step movement was significantly faster than 2-step. We recommend that the badminton players should practice 3-step forward footwork technique. The additional strength and power training for lower limbs should be carried out for the footwork training

Inhibition of Serine Protease Activity Protects Against High Fat Diet-Induced Inflammation and Insulin Resistance.

Recent evidence suggests that enhanced protease-mediated inflammation may promote insulin resistance and result in diabetes. This study tested the hypothesis that serine protease plays a pivotal role in type 2 diabetes, and inhibition of serine protease activity prevents hyperglycemia in diabetic animals by modulating insulin signaling pathway. We conducted a single-center, cross-sectional study with 30 healthy controls and 57 patients with type 2 diabetes to compare plasma protease activities and inflammation marker between groups. Correlations of plasma total and serine protease activities with variables were calculated. In an in-vivo study, LDLR-/- mice were divided into normal chow diet, high-fat diet (HFD), and HFD with selective serine protease inhibition groups to examine the differences of obesity, blood glucose level, insulin resistance and serine protease activity among groups. Compared with controls, diabetic patients had significantly increased plasma total protease, serine protease activities, and also elevated inflammatory cytokines. Plasma serine protease activity was positively correlated with body mass index, hemoglobin A1c, homeostasis model assessment-insulin resistance index (HOMA-IR), tumor necrosis factor-α, and negatively with adiponectin concentration. In the animal study, administration of HFD progressively increased body weight, fasting glucose level, HOMA-IR, and upregulated serine protease activity. Furthermore, in-vivo serine protease inhibition significantly suppressed systemic inflammation, reduced fasting glucose level, and improved insulin resistance, and these effects probably mediated by modulating insulin receptor and cytokine expression in visceral adipose tissue. Our findings support the serine protease may play an important role in type 2 diabetes and suggest a rationale for a therapeutic strategy targeting serine protease for clinical prevention of type 2 diabetes