Mutations in N-terminal flanking region of blue light-sensing light-oxygen and voltage 2 (LOV2) domain disrupt its repressive activity on kinase domain in the Chlamydomonas phototropin.

Phototropin is a light-regulated kinase that mediates a variety of photoresponses such as phototropism, chloroplast positioning, and stomata opening in plants to increase the photosynthetic efficiency. Blue light stimulus first induces local conformational changes in the chromophore-bearing light-oxygen and voltage 2 (LOV2) domain of phototropin, which in turn activates the serine/threonine (Ser/Thr) kinase domain in the C terminus. To examine the kinase activity of full-length phototropin conventionally, we employed the budding yeast Saccharomyces cerevisiae. In this organism, Ser/Thr kinases (Fpk1p and Fpk2p) that show high sequence similarity to the kinase domain of phototropins exist. First, we demonstrated that the phototropin from Chlamydomonas reinhardtii (CrPHOT) could complement loss of Fpk1p and Fpk2p to allow cell growth in yeast. Furthermore, this reaction was blue light-dependent, indicating that CrPHOT was indeed light-activated in yeast cells. We applied this system to a large scale screening for amino acid substitutions in CrPHOT that elevated the kinase activity in darkness. Consequently, we identified a cluster of mutations located in the N-terminal flanking region of LOV2 (R199C, L202L, D203N/G/V, L204P, T207I, and R210H). An in vitro phosphorylation assay confirmed that these mutations substantially reduced the repressive activity of LOV2 on the kinase domain in darkness. Furthermore, biochemical analyses of the representative T207I mutant demonstrated that the mutation affected neither spectral nor multimerization properties of CrPHOT. Hence, the N-terminal flanking region of LOV2, as is the case with the C-terminal flanking Jα region, appears to play a crucial role in the regulation of kinase activity in phototropin

Comparison between adhesion properties of adhesive bonding and adhesive-free adhesion for heat-assisted plasma-treated polytetrafluoroethylene (PTFE)

Heating during plasma treatment, known as heat-assisted plasma treatment, has recently reported to positively affect the adhesion properties of polytetrafluoroethylene (PTFE). In the present study, the adhesion properties of adhesive bonding and adhesive-free adhesion were compared for plasma-treated PTFE with different plasma treatment times and with or without heating during the plasma treatment. The relations among adhesion strength, plasma treatment time, radical density ratio, surface morphology, and surface hardness were investigated. No correlation was found between the adhesion strength and the radical density ratio or between the adhesion strength and the oxygen-containing-functional-group ratio. In contrast, correlation was observed between the adhesion strength and the surface hardness. In addition, the heat-assisted plasma treatment time affected the recovery of the weak boundary layer on the PTFE surface. Adhesive-free adhesion was found to require a longer heat-assisted plasma treatment time than adhesive bonding in order to achieve a high adhesion strength such as 1 N/mm

Effect of rubber compounding agent on adhesion strength between rubber and heat-assisted plasma-treated polytetrafluoroethylene (PTFE)

Although heat-assisted plasma treatment enables drastic improvement of the adhesion property of polytetrafluoroethylene (PTFE), plasma-treated PTFE does not strongly adhere to any adherend. To clarify which rubber compounding agents positively affect the adhesion strength of a plasma-treated PTFE/rubber assembly, six types of unvulcanised rubbers were prepared and thermally compressed to a plasma-treated PTFE sheet. Thus, it was found that SiO 2 addition to rubber drastically increased the adhesion strength of a plasma-treated PTFE/rubber assembly and cohesion failure of rubber occurred with large fractions of SiO 2 although no adhesives were used. To confirm the reaction between plasma-treated PTFE and SiO 2 powder, X-ray photoelectron spectroscopy (XPS) measurements were performed for the thermally compressed SiO 2 /PTFE assembly after repeated washing. The XPS results indicated that hydrophilic SiO 2 powder strongly adhered to the plasma-treated PTFE, whereas hydrophobic SiO 2 powder did not adhere to the PTFE. In this paper, a model was proposed for a possible mechanism of strong adhesion of a PTFE/rubber assembly through both hydrogen and covalent bonds between silanol groups of the SiO 2 powder surface in the rubber and hydroxyl or carboxyl groups on the plasma-treated PTFE

Drastic Improvement in Adhesion Property of Polytetrafluoroethylene (PTFE) via Heat-Assisted Plasma Treatment Using a Heater

Abstract The heating effect on the adhesion property of plasma-treated polytetrafluoroethylene (PTFE) was examined. For this purpose, a PTFE sheet was plasma-treated at atmospheric pressure while heating using a halogen heater. When plasma-treated at 8.3 W/cm2 without using the heater (Low-P), the surface temperature of Low-P was about 95 °C. In contrast, when plasma-treated at 8.3 W/cm2 while using the heater (Low-P+Heater), the surface temperature of Low-P+Heater was controlled to about 260 °C. Thermal compression of the plasma-treated PTFE with or without heating and isobutylene−isoprene rubber (IIR) was performed, and the adhesion strength of the IIR/PTFE assembly was measured via the T-peel test. The adhesion strengths of Low-P and Low-P+Heater were 0.12 and 2.3 N/mm, respectively. Cohesion failure of IIR occurred during the T-peel test because of its extremely high adhesion property. The surfaces of the plasma-treated PTFE with or without heating were investigated by the measurements of electron spin resonance, X-ray photoelectron spectroscopy, nanoindentation, scanning electron microscopy, and scanning probe microscopy. These results indicated that heating during plasma treatment promotes the etching of the weak boundary layer (WBL) of PTFE, resulting in a sharp increase in the adhesion property of PTFE

Progressively increased M50 responses to repeated sounds in autism spectrum disorder with auditory hypersensitivity: a magnetoencephalographic study.

The aim of this study was to investigate the differential time-course responses of the auditory cortex to repeated auditory stimuli in children with autism spectrum disorder (ASD) showing auditory hypersensitivity. Auditory-evoked field values were obtained from 21 boys with ASD (12 with and 9 without auditory hypersensitivity) and 15 age-matched typically developing controls. M50 dipole moments were significantly increased during the time-course study only in the ASD with auditory hypersensitivity compared with those for the other two groups. The boys having ASD with auditory hypersensitivity also showed more prolonged response duration than those in the other two groups. The response duration was significantly related to the severity of auditory hypersensitivity. We propose that auditory hypersensitivity is associated with decreased inhibitory processing, possibly resulting from an abnormal sensory gating system or dysfunction of inhibitory interneurons