Secondary-Ion Mass Spectrometry Images Cardiolipins and Phosphatidylethanolamines at the Subcellular Level

Millions of diverse molecules constituting the lipidome act as important signals within cells. Of these, cardiolipin (CL) and phosphatidylethanolamine (PE) participate in apoptosis and ferroptosis, respectively. Their subcellular distribution is largely unknown. Imaging mass spectrometry is capable of deciphering the spatial distribution of multiple lipids at subcellular levels. Here we report the development of a unique 70 keV gas-cluster ion beam that consists of (CO₂)_n⁺(n>10 000) projectiles. Coupled with direct current beam buncher-time-of-flight secondary-ion mass spectrometry, it is optimized for sensitivity towards high-mass species (up to m/z 3000) at high spatial resolution (1 μm). In combination with immunohistochemistry, phospholipids, including PE and CL, have been assessed in subcellular compartments of mouse hippocampal neuronal cells and rat brain tissue.     

流体力学半径对估算胶体微球聚集速率常数的影响

胶体粒子聚集速率常数实验值远低于理论值一直是被普遍关注的问题.聚集速率常数的理论推导是基于粒子的几何半径来考虑的,但决定粒子扩散速率及聚集速率的应该是粒子的流体力学半径(大于几何半径),因而它是使聚集速率常数实验值低于理论值的因素之一.影响流体力学半径的因素很多,其中,带电粒子在溶液中因表面存在双电层,会明显增大流体力学半径,造成聚集速率减慢.而双电层的厚度又随溶液中离子强度的不同而改变.本工作在聚集速率的公式中引入了修正因子,即几何半径与其流体力学半径之比,以修正由于用几何半径代替流体力学半径带来的误差.其中几何半径和流体力学半径可以分别用扫描电镜(SEM)和动态光散射(DLS)来测定.以两种粒径的聚苯乙烯带电微球为例,考察了在不同离子强度下,该误差的大小.结果发现,对于半径为30 nm的微球,用流体力学半径计算的慢聚集速率常数比理论值偏低约8%.该误差随离子强度增加而减少.对于快聚集情况,流体力学半径对聚集速率基本没有影响.     

Nitroxides scavenge myeloperoxidase-catalyzed thiyl radicals in model systems and in cells

Nitroxide radicals possess important antioxidant activity in live tissues because of their ability to scavenge reactive radicals. Despite the fact that, in cells, damaging free radicals are primarily quenched by glutathione (GSH) with subsequent formation of harmful glutathionyl radical (GS(*)), interactions of nitroxide radicals with GS(*) and thiols have not been studied in detail. In addition, intracellular metabolic pathways leading to the formation of secondary amines from nitroxides are unknown. Here we report that GS(*) radicals react efficiently and irreversibly with nitroxides to produce secondary amines. We developed a sensitive method for the detection of GS(*) based on their specific interaction with Ac-Tempo, a nonfluorescent conjugate of fluorogenic acridine with paramagnetic nitroxide Tempo, and used it to characterize interactions between nitroxide and thiyl radicals generated through phenoxyl radical recycling by peroxidase. During reaction of Ac-Tempo with GS(*), Tempo EPR signals decayed and acridine fluorescence concurrently increased. DMPO and PBN, spin traps for GS(*), inhibited this interaction. Using combined HPLC and mass spectrometry, we determined that 90% of the Ac-Tempo was converted into fluorescent acridine (Ac)-piperidine; GSH was primarily oxidized into sulfonic acid. In myeloperoxidase-rich HL-60 cells, Ac-piperidine fluorescence was observed upon stimulation of GS(*) generation by H(2)O(2) and phenol. Development of fluorescence was prevented by preincubation of cells with the thiol-blocking reagent N-ethylmaleimide as well as with peroxidase inhibitiors. Furthermore, Ac-Tempo preserved intracellular GSH and protected cells from phenol/GS(*) toxicity, suggesting a new mechanism for the free-radical scavenging activity of nitroxides in live cells.     

Mitochondrial targeting of radioprotectants using peptidyl conjugates

Ionizing radiation activates a mitochondrial nitric oxide synthase, leading to inhibition of the respiratory chain, generation of excess superoxide, peroxynitrite production and nitrosative damage. We have measured the radioprotective effects of a nitric oxide synthase antagonist (AMT) versus a free radical scavenger (4-amino-TEMPO) using electrochemical detection of nitric oxide and peroxynitrite. To enhance their efficacy, we have conjugated these compounds to peptides and peptide isosteres--derived from the antibiotic gramicidin S--that target the mitochondria. The targeting ability of these peptidyl conjugates was measured using quantitative mass spectrometry.     

Secondary‐Ion Mass Spectrometry Images Cardiolipins and Phosphatidylethanolamines at the Subcellular Level

Millions of diverse molecules constituting the lipidome act as important signals within cells. Of these, cardiolipin (CL) and phosphatidylethanolamine (PE) participate in apoptosis and ferroptosis, respectively. Their subcellular distribution is largely unknown. Imaging mass spectrometry is capable of deciphering the spatial distribution of multiple lipids at subcellular levels. Here we report the development of a unique 70 keV gas‐cluster ion beam that consists of (CO₂)_n⁺(n>10 000) projectiles. Coupled with direct current beam buncher‐time‐of‐flight secondary‐ion mass spectrometry, it is optimized for sensitivity towards high‐mass species (up to m/z 3000) at high spatial resolution (1 μm). In combination with immunohistochemistry, phospholipids, including PE and CL, have been assessed in subcellular compartments of mouse hippocampal neuronal cells and rat brain tissue.     

Peak effect studies in single crystals CeRu2 and 2H-NbS2

We have studied the peak effect (PE) phenomenon in single crystals of weakly pinned superconductors CeRu₂ and 2H-NbS₂. 2H-NbS₂ is iso-structural and iso-electronic to 2H-NbSe₂, whose similarity with CeRu₂ as regards the PE representing the order-to-disorder transformation of the flux line lattice was claimed some time ago. We report on the step change in equilibrium magnetization across the peak effect in CeRu₂. We also present the vortex phase diagram of 2H-NbS₂ obtained from the magnetization data, and compare the PE phenomenon in 2H-NbS₂ and 2H-NbSe₂.     

Temperature hysteretic effect and its influence on colossal magnetoresistance of La0.33Nd0.33Ca0.33MnO3

Electrical resistance (R) measurements of a bulk La_0.33Nd_0.33Ca_0.33MnO₃ perovskite in magnetic fields up to 40 kOe have revealed anomalous temperature hysteretic effects both in 0 Oe and 20 kOe magnetic fields. The sharp peak observed in the R vs. T plot indicates the occurrence of metal-to-insulator (M-I) transition at a temperature of T _MI=110 K and 140 K, for cooling and warming paths, respectively. An applied magnetic field of 20 kOe reduces the resistance and shifts T _MI to 160 K and 185 K for cooling and warming, respectively. We have observed a much higher resistance in the cooling path than in the warming path leading to the hysteretic resistance ratio (R _cool/R _warm) of 200 at 110 K and 1.8 at 160 K for 0 Oe and 20 kOe, respectively. Record values of colossal magnetoresistance (CMR) have been achieved. The CMR value reaches nearly 99% in the temperature ranges of 90 K to 140 K and 90 K to 170 K for 20 kOe and 40 kOe magnetic fields in the cooling mode, respectively. The observed unusual behavior is attributed to the co-existence of La-rich and Nd-rich domains assumed to be distributed randomly in the compound.     

Synthetic magnetic opals

We present studies of novel nanocomposites of BiNi impregnated into the structure of opals as well as inverse opals. Atomic force microscopy and high resolution elemental analyses show a highly ordered structure and uniform distribution of the BiNi filler in the matrix. These BiNi-based nanocomposites are found to exhibit distinct ferromagnetic-like ordering with transition temperature of about 675 K. As far as we know there exists no report in literature on any BiNi compound which is magnetic.