Tyrosine kinase, phosphatidylinositol 3-kinase, and protein kinase C regulate insulin-stimulated NaCl absorption in the thick ascending limb

Tyrosine kinase, phosphatidylinositol 3-kinase, and protein kinase C regulate insulin-stimulated NaCl absorption in the thick ascending limb. We have previously shown a direct stimulatory effect of insulin on NaCl absorption in the medullary thick ascending limb of Henle's loop (mTAL). To further investigate the signal transduction involved, we determined whether tyrosine kinase, phosphatidylinositol 3-kinase (PI3-kinase), and/or protein kinase C (PKC) regulate insulin-stimulated NaCl absorption in the mTAL by in vitro microperfusion methods. In control experiments, insulin increased transepithelial voltage (Vte) and net lumen-to-bath Cl− flux (JCl). Genistein and methyl 2,5-dihydroxycinnamate, two specific tyrosine kinase inhibitors, abolished the effects of insulin. Wort-mannin, a specific PI3-kinase inhibitor, inhibited the action of insulin. The effects of insulin also were inhibited by staurosporin and calphostin C, which are dissimilar inhibitors of PKC. These results indicate that insulin stimulates NaCl absorption in the mTAL through tyrosine kinase, PI3-kinase, and PKC-mediated mechanisms. Moreover, because we have reported previously that insulin causes no detectable change in cytosolic free Ca2+ in the mTAL cells, the present results also suggest that insulin-induced PKC activation is not related to inositol 1,4,5-triphosphate (IP3) production

Pathogenetic roles of beet necrotic yellow vein virus RNA5 in the exacerbation of symptoms and yield reduction, development of scab‐like symptoms, and Rz1‐resistance breaking in sugar beet

Beet necrotic yellow vein virus (BNYVV) generally has a four‐segmented positive‐sense RNA genome (RNAs 1–4), but some European and most Asian strains have an additional segment, RNA5. This study examined the effect of RNA5 and RNA3 on different sugar beet cultivars using a Polymyxa‐mediated inoculation system under field and laboratory conditions. In field tests, the degree of sugar yield served as an index for assessing the virulence of BNYVV strains. Japanese A‐II type isolates without RNA5 caused mostly 15%–90% sugar yield reductions, depending on the susceptibility of sugar beet cultivars, whereas the isolates with RNA5 induced more than 90% yield losses in the seven susceptible cultivars, but small yield losses in one Rz1‐resistant and Rizor cultivars. However, a laboratory‐produced isolate containing RNA5 but lacking RNA3 caused higher yield losses in Rizor than in susceptible plants, and induced scab‐like symptoms on the root surface of both susceptible and resistant plants. In laboratory tests, A‐II type isolates without RNA5 had low viral RNA accumulation levels in roots of Rizor and Rz1‐resistant plants at early stages of infection, but in the presence of RNA5, viral RNA3 accumulation levels increased remarkably. This increased RNA3 accumulation was not observed in roots of the WB42 accession with the Rz2 gene. In contrast, the presence of RNA3 did not affect RNA5 accumulation levels. Collectively, this study demonstrated that RNA5 is involved in the development of scab‐like symptoms and the enhancement of RNA3 accumulation, and suggests these characteristics of RNA5 are associated with Rz1‐resistance breaking

Pharmacokinetics of Beclomethasone Dipropionate in an Hydrofluoroalkane-134a Propellant System in Japanese Children with Bronchial Asthma

ABSTRACTBackgroundHydrofluoroalkane-134a (HFA) has been shown to be a safe replacement for chlorofluorocarbons (CFCs) as a pharmaceutical propellant, with the advantage that it has no ozone-depleting potential. This is the first report of the pharmacokinetics of beclomethasone dipropionate (BDP) delivered from a pressurized solution formulation using an HFA propellant system (HFA-BDP) in Japanese children with bronchial asthma.MethodsPlasma concentrations of beclomethasone 17-monopropionate (17-BMP), a major metabolite of BDP, following an inhaled dose of HFA-BDP (200 μg as four inhalations from 50 μg/actuation) in five Japanese children with bronchial asthma were quantified and analyzed by a non-compartmental analysis to obtain pharmacokinetic parameters.ResultsThe area under the concentration-time curve from time zero to the last quantifiable time (AUC0-t) was 1659 ± 850 pg • h/mL (arithmetic mean ± standard deviation (SD)), the maximum concentration observed (Cmax) was 825 ± 453 pg/mL and the apparent elimination half-life (t1/2) was 2.1 ± 0.7 hours. The time to reach Cmax (Tmax) was 0.5 hours in all patients. No special relationship was observed between these parameters and age or body weight. These parameters were compared with the previously reported parameters of American children with bronchial asthma. The Japanese/American ratio of the geometric means of each parameter was 1.36 for AUC0-t, 1.04 for Cmax and 1.4 for t1/2. The median of Tmax was 0.5 hours in American patients as well as Japanese patients.ConclusionsThe pharmacokinetics of HFA-BDP in Japanese children with bronchial asthma are reported for the first time and a similarity to those in American children is suggested

Pathogenic mutations identified by a multimodality approach in 117 Japanese Fanconi anemia patients

Fanconi anemia is a rare recessive disease characterized by multiple congenital abnormalities, progressive bone marrow failure, and a predisposition to malignancies. It results from mutations in one of the 22 known FANC genes. The number of Japanese Fanconi anemia patients with a defined genetic diagnosis was relatively limited. In this study, we reveal the genetic subtyping and the characteristics of mutated FANC genes in Japan and clarify the genotype-phenotype correlations. We studied 117 Japanese patients and successfully subtyped 97% of the cases. FANCA and FANCG pathogenic variants accounted for the disease in 58% and 25% of Fanconi anemia patients, respectively. We identified one FANCA and two FANCG hot spot mutations, which are found at low percentages (0.04-0.1%) in the whole-genome reference panel of 3,554 Japanese individuals (Tohoku Medical Megabank). FANCB was the third most common complementation group and only one FANCC case was identified in our series. Based on the data from the Tohoku Medical Megabank, we estimate that approximately 2.6% of Japanese are carriers of disease-causing FANC gene variants, excluding missense mutations. This is the largest series of subtyped Japanese Fanconi anemia patients to date and the results will be useful for future clinical management

Ultrafast Excited State Reaction Dynamics in Aqueous Nanodroplets and Fluorescent Proteins

EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Anisotropic defect distribution in He+-irradiated 4H-SiC: Effect of stress on defect distribution

Irradiation-induced anisotropic swelling in hexagonal alpha-SiC is known to degrade the mechanical properties of SiC; however, the associated physical mechanism and microstructural process remain insufficiently understood. In this study, an anisotropic swelling condition where the surface normal direction was allowed to freely expand with constraint in the lateral direction was introduced in 4H-SiC using selected-area He+ irradiation, and the internal defect distribution was investigated using transmission electron microscopy (TEM) and advanced scanning TEM. The defect distribution was compared to that in non-selected-area He+-irradiated 4H-SiC and electron-irradiated TEM-foil 4H-SiC. An anisotropic defect distribution was observed in the selected-area He+-ion-irradiated 4H-SiC, with interstitial defects preferentially redistributed in the surface normal direction ([0 004]) and negative volume defects (such as vacancies and/or carbon antisite defects) dominantly located in the lateral directions ([11 (2) over bar0] and [10 (1) over bar0]). This anisotropy of the defect distribution was substantially lower in the non-selected-area He+-irradiated and electron-irradiated samples. The stress condition in the three samples was also measured and analyzed. In the selected-area He+-irradiated 4H-SiC, compressive stress was introduced in the lateral directions (([10 (1) over bar0] and [11 (2) over bar0])), with little stress introduced in the surface normal direction ([0 004]); this stress condition was introduced at the beginning of ion irradiation. The compressive stress likely inhibits the formation of interstitial defects in the lateral directions, enhancing the anisotropy of the defect distribution in SiC

Electron energy-loss spectroscopic evaluation of depth-dependent swelling of He+ ion-irradiated 4H-SiC correlated with defect type

Various defects and amorphous transitions are the primary mechanism behind the accumulation of swelling in silicon carbide (SiC). In this study, selected-area He+ ion irradiation was carried out on single-crystal 4H-SiC using fluences of 1x10(15), 5x10(16), and 1x10(17)cm(-2) at room temperature. The defect distribution in the samples with varying irradiation fluences was analyzed using transmission electron microscopy (TEM), while the local swelling of regions under varying damage conditions was estimated using electron energy-loss spectroscopy. The results provide the range of swelling in SiC possessing different primary defect types, such as point defects or tiny clusters, black spot defects, and amorphous SiC. A saturation swelling with a value of 2%-3% in the near-surface region, induced by point defects or tiny clusters (invisible in TEM), was observed at room temperature over the fluence range of 1x10(15) to 1x10(17)cm(-2). This saturation has already reached at a great low dose of about 0.02dpa. The swelling of the region containing black spot defects ranges from about 3% to 7%. Helium bubbles increase the volume swelling of SiC, while the He+ ion irradiation may also perform a decreasing effect on the volume swelling below a certain irradiation fluence

Non-destructive evaluation of the strain distribution in selected-area He+ ion irradiated 4H-SiC

Residual strain in silicon carbide (SiC) greatly affects its physical and chemical properties and thus the performance of SiC-based devices. Herein, the detailed strain distribution in selected-area He+ ion-irradiated 4H-SiC was evaluated using the non-destructive techniques of electron backscattering diffraction and confocal Raman microscopy (CRM). In addition to the strain introduced in the irradiated area, excessive strain induced by irradiation-induced swelling also extended into the surrounding substrate. Furthermore, great compressive strain was concentrated around the interface between the irradiated and unirradiated areas. In the strain-introduced substrate, an A(1)(LO)/A(1)(LOPC) peak variation was detected by CRM, suggesting a variation of the carrier density