Fast X-ray fluorescence microscopy provides high-throughput phenotyping of element distribution in seeds.

The concentration, chemical speciation, and spatial distribution of essential and toxic mineral elements in cereal seeds have important implications for human health. To identify genes responsible for element uptake, translocation, and storage, high-throughput phenotyping methods are needed to visualize element distribution and concentration in seeds. Here, we used X-ray fluorescence microscopy (µ-XRF) as a method for rapid and high-throughput phenotyping of seed libraries and developed an ImageJ-based pipeline to analyze the spatial distribution of elements. Using this method, we nondestructively scanned 4,190 ethyl methanesulfonate (EMS)-mutagenized M1 rice (Oryza sativa) seeds and 533 diverse rice accessions in a genome-wide association study (GWAS) panel to simultaneously measure concentrations and spatial distribution of elements in the embryo, endosperm, and aleurone layer. A total of 692 putative mutants and 65 loci associated with the spatial distribution of elements in rice seed were identified. This powerful method provides a basis for investigating the genetics and molecular mechanisms controlling the accumulation and spatial variations of mineral elements in plant seeds.

Nutrient accumulation and transcriptome patterns during grain development in rice.

Rice is an important source of calories and mineral nutrients for more than half of the world's population. The accumulation of essential and toxic mineral elements in rice grain affects its nutritional quality and safety. However, the patterns and processes by which different elements progressively accumulate during grain filling remain largely unknown. In the present study, we investigated temporal changes in dry matter, elemental concentrations, and the transcriptome in the grain of field-grown rice. We also investigated the effects of seed setting rate and the position of the grain within the rice panicle on element accumulation. Three different patterns of accumulation were observed: (i) elements including K, Mn, B, and Ca showed an early accumulation pattern; (ii) dry matter and elements including N, P, S, Mg, Cu, Zn, Mo, As, and Cd showed a mid accumulation pattern; and (iii) elements such as Fe showed a gradual increase pattern. These different accumulation patterns can be explained by the differences in the biogeochemical behavior of the various elements in the soil, as well as differences in plant nutrient redistribution, gene expression, and the sink-source relationship. These results improve our knowledge of the dynamics of elemental accumulation in rice grain and are helpful for identification of functional genes mediating the translocation of elements to grain.

Feasibility of Atrial AutoCapture™ to Detect Atrial Evoked Response: Experience from 102 Patients Implanted with Dual-chamber Pacemakers.

BACKGROUND: Atrial AutoCapture™ (ACap™) was a new technological development that confirmed atrial capture by analyzing evoked response (ER) with a new method - paced depolarization integral ER detection - and optimized energy output to changes in the stimulation threshold. The purpose of this study was to evaluate the clinical performance of ACap™ function. METHODS: This was a prospective, observational, nonrandomized two-center study. Between November 2008 and August 2014, 102 patients were enrolled from two different institutions. Data were collected by case report forms at enrollment, hospital discharge, and in-office follow-ups scheduled at 1, 2, 3, 6, and 12 months postimplantation. RESULTS: Ambulatory ACap™ function started to become available for 20.6% of patients at 1 day, then progressed to 30.4% at 7 days, 38.6% at 1 month, 41.6% at 2 months, 47.5% at 3 months, 53.5% at 6 months, and 63.4% at 1 year. The cause of the unsuccessful attempts to perform ACap™ threshold was ER/polarization <2:1. Availability for SD, BND, and HOCM indications had shown better results than AVB indication. For SD indication cases, feasibility was significantly better for SD with paroxysmal atrial fibrillation (pAF) than SD without pAF (78.4% vs. 35.0% at 1 year, n = 71, P< 0.001). At each stage of the clinical follow-ups, there had been a strict correlation between ACap™ measurements and those conducted manually with P 0.001 (n = 299). CONCLUSIONS: It has been concluded that ACap™ function was safe and effective to confirm atrial threshold and reduce energy output automatically. ACap™ function is unavailable for some patients at early stages of the implantation; however, availability has been progressively increasing during follow-up.

[Effects of oxidized-low density lipoprotein on expression of urotensin II receptor GPR14 in rat aortic smooth muscle cells].

OBJECTIVE: To investigate the effects of oxidized-low density lipoprotein (ox-LDL) and urotensin II (U II) on the proliferation of rat aortic smooth muscle cells (VSMCs) and the effect of ox-LDL on the mRNA expression of U II receptor GPR14 in. METHODS: Rat VSMCs were incubated with UII and ox-LDL at different concentrations. The viability of rat VSMCs was detected by MTT assay. Rat VSMCs were incubated with ox-LDL at the concentrations of 0, 0.1, 1, 10, and 50 microg/ml. Twelve hours later competitive RT-PCR was used to detect the effects of concentration of ox-LDP on the mRNA expression of GPR14 in the VSMCs. Another VSMCs were incubated with ox-LDL at the final concentration of 50 microg/ml, and 0, 2, 6, 12, and 24 hours later competitive RT-PCR was used to detect the mRNA expression of GPR14 in the VSMCs. (125)I-labelled U II was added into the culture fluid of VSMCs, and radioligand binding assay was used to calculate the maximum binding (B(max)). RESULTS: UII of the concentration of 50 nmol/L significantly increased the proliferation of VSMCs. When the concentration of ox-LDL was 10 microg/ml the proliferation of VSMCs (P < 0.01) was 0.678 +/- 0.061, increased by 121% compared with the control group (0.325 +/- 0.052, P < 0.01). 0.01 microg/ml of ox-LDL and 10 nmol/L of UII showed a synergistic effect on the proliferation of VSMCs with the MTT value increased to 162% +/- 29% that of the control group (P < 0.01). When the concentration of ox-LDL increase to 0.1 microg/ml the synergistic effect was attenuated with the MTT value of 153% +/- 22% that of the control group (P < 0.05) and when the concentration of ox-LDL increased to 1 microg/ml the synergistic effect diminished, with a MTT value of 123% +/- 13% that of the control group (P > 0.05). When the concentration of ox-LDL was 50 microg/ml the proliferation of VSMCs the MTT value decreased to 59% that of the of control group, however, ox-LDL of the concentration of 50 microg/ml combined with UII of the concentration of 10 nmol/L the MTT value increased to 68% that of the control group, showing that UII of that concentration significantly inhibited the cytotoxic effect of ox-LDL. Incubation of VSMCs with the ox-LDL at the concentration of 0.1 microg/ml the GPR14 mRNA expression of the VSMCs was 125.1% that of the control group (P < 0.01) as assessed by competitive RT-PCR, however, when the concentration of ox-LDL increased over 1 microg/ml the GPR14 mRNA expression does-dependently decreased as much as 72.6% that of the control group (all P < 0.01), with the maximal effect when the concentration of ox-LDL reached 50 microg/ml. ox-LDL at the concentration of 50 microg/ml time-dependently down-regulated the GPR14 mRNA expression with the maximal effect 12 hours after addition of ox-LDL and this effect was sustained for up to 24 hours. After incubation for 12 hours the binding of (125)I-UII to VSMCs was 117.5% that of the control group (P < 0.01) with the ox-LDL at the concentration of 0.1 microg/ml, and was 68.8% that of the control group (P < 0.01) with the ox-LDL at the concentration of 50 microg/ml. CONCLUSION: ox-LDL and with U-II. GPR14 at certain concentrations show synergetic effects on the proliferation of VSMCs. The GPR14 mRNA expression in VSMCs can be upregulated by low concentration of ox-LDL, while downregulated by high concentration of ox-LDL.

[The pulmonary toxicity of amiodarone: six-case report].

OBJECTIVE: To report the clinical features and the prognosis, the methods of diagnosis and treatment, and the early detection of the pulmonary toxicity induced by amiodarone. METHOD: The clinical course, the findings of X-ray and CT and the results of treatment were summarized and analyzed in six patients with amiodarone-induced pulmonary toxicity. RESULTS: Five males, one female, aged 62 - 69 (66.0 +/- 2.4) years. Amiodarone was used because of paroxysmal atrial fibrillation in five patients and ventricular arrhythmia in one. The loading dose was 7 g and the maintaining dose was 0.2 g/d or less. Pulmonary toxicity was recognized at the times in 0.5 - 4.0 (2.1 +/- 1.3) years after amiodarone therapy. Dyspnea occurred, crack rales were audible in both lower parts of the lungs, and the chest X-ray showed grid-like changes in case one. No symptom was found in the others. Their diagnosis was made according to the pulmonary intestinal changes by high-resolution computerized tomography when the lung marking was increased or deranged by chest X-ray during the regular follow-up. Pulmonary function examination showed that the restrictive ventilation and the CO diffusing capacity decreased in case one, while the CO diffusing capacity was normal in the others. The decreased obstructive ventilation capacity was found in case six. Amiodarone was discontinued in all the cases after the diagnosis of induced pulmonary toxicity. One patient was treated with corticosteroid, three with azithromycin, and the another two patients were not treated with drug. During 0.1 - 5.0 year period of follow-up the symptoms were markedly attenuated in case one, and no new symptoms and radiography findings were found in the others. CONCLUSIONS: Pulmonary toxicity is a serious adverse effect of amiodarone. The typical feature is pulmonary intestinal fibrosis and thick pulmonary intestine in the early stage. Corticosteroid treatment seems effective. It would be helpful for early diagnosis to take chest X-ray examination regularly and CT examination in suspicions cases during the therapy. The prognosis may be good in the early diagnosed cases.

[Effect of different isoforms of tocopherols on expression of intercellular adhesion molecule-1 in human umbilical vein endothelial cells].

OBJECTIVE: To observe the influence of different tocopherol isoforms on oxidized low density lipoprotein (oxLDL) or recombinant human C-reactive protein (rhCRP)-induced expression of intercellular adhesion molecule-1 (ICAM-1) in human umbilical vein endothelial cells (HUVECs) and to investigate the potential mechanisms and effects of different tocopherols on atherosclerosis. METHODS: Cultured HUVECs were incubated with oxLDL, oxLDL + alpha-tocopherol, oxLDL + gamma-tocopherol, oxLDL + mixed-tocopherols, rhCRP, rhCRP + alpha-tocopherol, rhCRP + gamma-tocopherol rhCRP + mixed-tocopherols for 24 hours, respectively. The ICAM-1 expressions of protein and mRNA were detected by cell enzyme linked immunosorbent assay (ELISA), flow cytometric technique and RT-PCR. RESULTS: Incubation of HUVECs with oxLDL or rhCRP for 24 hours significantly increased ICAM-1 expressions of proteins and mRNA. The different tocopherols inhibited oxLDL-induced ICAM-1 expression in HUVECs in a concentration-dependent manner (50-200 micromol/L) and mixed-tocopherols were more potent than alpha-tocopherol or gamma-tocopherol alone. However, rhCRP-induced ICAM-1 expression in HUVECs was not inhibited by tocopherols. CONCLUSION: The different tocopherols inhibited oxLDL-induced ICAM-1 expression in HUVECs and mixed-tocopherols were more potent than alpha-tocopherol or gamma-tocopherol alone, which may be important for the beneficial effects of tocopherols on atherosclerosis and cardiovascular disease.