Modulation of viability of live cells by focused ion‐beam exposure

Introduction of membrane‐impermeant substances into living cells is the key method to understand contemporary cellular processes by investigating cellular responses and phenotypes. Here, we performed gold ion beam exposure into live cells by using the focused ion beam implantation method, which was originally developed to precisely control semiconductor device performances. We evaluated the viability of the gold‐irradiated cells by measuring the concentration of adenosine triphosphate (ATP), which is an intracellular energy source produced in the mitochondrial membrane. The viability of the irradiated cells was found to be 20% higher than that of the unirradiated control cells. The atoms might promote the energy generating processes within the mitochondrion. Our results suggest that the viability of living cells can be modulated by accurately controlling the dopant atom numbers. Our technique may be considered as a potential tool in life and medical sciences to quantitatively elucidate the dose‐dependent effects of dopants. Biotechnol. Bioeng. 2011; 108:222–225. © 2010 Wiley Periodicals, Inc.     

Submillisecond Conformational Changes in Proteins Resolved by Photothermal Beam Deflection

Photothermal beam deflection together with photo-acoustic calorimetry and thermal grating belongs to the family of photothermal methods that monitor the time-profile volume and enthalpy changes of light induced conformational changes in proteins on microsecond to millisecond time-scales that are not accessible using traditional stop-flow instruments. In addition, since overall changes in volume and/or enthalpy are probed, these techniques can be applied to proteins and other biomacromolecules that lack a fluorophore and or a chromophore label. To monitor dynamics and energetics of structural changes associated with Ca²⁺ binding to calcium transducers, such neuronal calcium sensors, a caged calcium compound, DM-nitrophen, is employed to photo-trigger a fast (τ < 20 μsec) increase in free calcium concentration and the associated volume and enthalpy changes are probed using photothermal beam deflection technique.     

Changes in DNA base sequences in the mutant of Arabidopsis thaliana induced by low-energy N+ implantation

To reveal the mutation effect of low-energy ion implantation on Arabidopsis thaliana in vivo, T80II, a stable dwarf mutant, derived from the seeds irradiated by 30 keV N⁺ with the dose of 80×10¹⁵ ions/cm² was used for Random Amplified Polymorphic DNA (RAPD) and base sequence analysis. The results indicated that among total 397 RAPD bands observed, 52 bands in T80II were different from those of wild type showing a variation frequency 13.1%. In comparison with the sequences of A. thalianain GenBank, the RAPD fragments in T80II were changed greatly in base sequences with an average rate of one base change per 16.8 bases. The types of base changes included base transition, transversion, deletion and insertion. Among the 275 base changes detected, single base substitutions (97.09%) occurred more frequently than base deletions and insertions (2.91%). And the frequency of base transitions (66.55%) was higher than that of base transversions (30.55%). Adenine, thymine, guanine or cytosine could be replaced by any of other three bases in cloned DNA fragments in T80II. It seems that thymine was more sensitive to the irradiation than other bases. The flanking sequences of the base changes in RAPD fragments in T80II were analyzed and the mutational “hotspot” induced by low-energy ion implantation was discussed.     

低剂量率下高线性能量转移碳离子束辐照人类唾液腺细胞的存活效应

利用日本千叶重离子医用加速器 HIMAC 提供的碳离子束,对人类唾液腺细胞 (HSG) 在剂量率为 0.5 Gy/h 的低剂量率条件下进行了辐照,运用标准的克隆形成法得到了 3 种不同剂量平均线性能量转移 (LET) 碳离子束辐照 HSG 细胞的剂量存活效应 . 与先前 HSG 细胞在治癌剂量率 (1~5 Gy/min) 下对相近剂量平均 LET 碳离子束辐照的剂量存活效应数据相比, HSG 细胞对高 LET 碳离子束辐射表现出明显的剂量率效应 . 为在相同条件下得到碳离子束对 HSG 细胞的相对生物学效应 (RBE) ,利用 60Co-γ射线在剂量率为 0.5 Gy/h 的条件下辐照了 HSG 细胞,得到该细胞系对低 LET 射线响应的剂量存活效应 . 与先前在治癌剂量率下得到的 RBE 值相比,低剂量率条件下得到的 RBE 值总体减小 . 由实验发现的剂量率效应及低剂量率条件下 RBE 值的减小,表明由高 LET 碳离子束造成的辐射损伤在低剂量率条件下也存在着显著的修复效应 . 据此,对辐射造成细胞致死的原因进行了探讨 .     

Fabrication and Operation of a Nano-Optical Conveyor Belt

The technique of using focused laser beams to trap and exert forces on small particles has enabled many pivotal discoveries in the nanoscale biological and physical sciences over the past few decades. The progress made in this field invites further study of even smaller systems and at a larger scale, with tools that could be distributed more easily and made more widely available. Unfortunately, the fundamental laws of diffraction limit the minimum size of the focal spot of a laser beam, which makes particles smaller than a half-wavelength in diameter hard to trap and generally prevents an operator from discriminating between particles which are closer together than one half-wavelength. This precludes the optical manipulation of many closely-spaced nanoparticles and limits the resolution of optical-mechanical systems. Furthermore, manipulation using focused beams requires beam-forming or steering optics, which can be very bulky and expensive. To address these limitations in the system scalability of conventional optical trapping our lab has devised an alternative technique which utilizes near-field optics to move particles across a chip. Instead of focusing laser beams in the far-field, the optical near field of plasmonic resonators produces the necessary local optical intensity enhancement to overcome the restrictions of diffraction and manipulate particles at higher resolution. Closely-spaced resonators produce strong optical traps which can be addressed to mediate the hand-off of particles from one to the next in a conveyor-belt-like fashion. Here, we describe how to design and produce a conveyor belt using a gold surface patterned with plasmonic C-shaped resonators and how to operate it with polarized laser light to achieve super-resolution nanoparticle manipulation and transport. The nano-optical conveyor belt chip can be produced using lithography techniques and easily packaged and distributed.     

Occurrence of the pugnose pipefish Bryx dunckeri in the Sargasso Sea

Juvenile specimens of the pugnose pipefish, Bryx dunckeri, were collected during a multipurpose research survey conducted within the Sargasso Sea Subtropical Convergence Zone, extending the known distribution range of this species to include open ocean areas of the Western North Atlantic. Novel spatial data are of scientific interest as information on the distribution, population structure, and population size of this species is limited. Additionally, we present detailed photographs and morphological data on the collected specimens. The results are discussed in relation to the dispersal abilities and population structure in syngnathids.     

Importance of discards of a beam trawl fishery as input of organic matter into nursery areas within the Tagus estuary

The brown-shrimp beam trawl fishery carried out within the Tagus estuary produces discards due to the little commercial interest of most of the species caught. Between 1994 and 1996, monthly samples were collected in the two major fishing areas within the Tagus estuary, using a beam trawl, in order to estimate the amount of fish and crustaceans caught per unit of effort. The fishing effort of the commercial fleet was determined based on surveys of professional fishermen. Mortality estimates of discards were also evaluated experimentally. The main fish and crustacean species discarded after capture were Crangon crangon (Linnaeus, 1758), Liza ramada (Risso, 1826), Carcinus maenas (Linnaues, 1758) and Pomatoschistus minutus (Pallas, 1770). The estimated total amount of fishery discards in the upper part of the Tagus estuary is approximately 1500 tonnes per year, which represents ca. 90% of the captures. The mortality rate of the fishes and crustaceans discarded varied according to species and season, with the highest rates during Summer months. Considering nitrogen and carbon content of the main discarded species, an input of particulate organic matter of more than 140 tonnes of carbon and 35 tonnes of nitrogen per year were estimated for these estuarine areas.     

CBCT引导放疗摆位误差对中上段食管癌患者受照射剂量的影响

摘要 目的：探究锥形束CT（CBCT）引导放疗摆位误差对中上段食管癌患者受照射剂量的影响。方法：选取2017年5月～2019年5月于我院收治的60例中上段食管癌患者为研究对象,所有患者均行CBCT图像、计划CT图像采集。在患者放疗前进行CBCT扫描,将CBCT图像与计划CT图像匹配,得到左右（x轴）、头脚（y轴）、前后（z轴）三个方向的线性误差,分析出现的误差及误差的分布规律。利用模拟实际照射系统,进行模拟计划,得到实际照射靶区及正常组织受照射剂量,将其与治疗前计划比较,研究摆位误差对患者受照剂量的影响。结果：患者整体摆位误差为x轴（2.91±2.20）mm,y轴（3.89±2.17）mm,z轴（2.44±1.64）mm,x轴的MPTV为4.054 mm,y轴的MPTV为8.183 mm,z轴的MPTV为3.482 mm。模拟计划的CI、PTV的Dmin、Dmean、D95%均低于标准计划差异显著（P均＜0.05）,而模拟计划的HI低于标准计划（P＜0.05）。模拟计划的脊髓Dmax高于标准计划（P＜0.05）,而标准计划与模拟计划的双肺V20、Dmean,心脏V40差异比较无统计学意义（P均＞0.05）。结论：CBCT引导放疗摆位误差对中上段食管癌患者影响较小,提高PTV受照射剂量及治疗准确程度,对脊髓有保护效果。摆位误差对心、肺的剂量分布无明显影响。     

Using focus ion beam to prepare crystal lamella for electron diffraction

Electron diffraction provides a powerful tool to solve the structures of small protein crystals. However, strong interactions between the electrons and the materials limit the application of the electron crystallographic method on large protein crystals with micrometer or larger sizes. Here, we used the focused ion beam (FIB) equipped on the scanning electron microscope (SEM) to mill a large crystal to thin lamella. The influences of the milling on the crystal lamella were observed and investigated, including radiation damage on the crystal surface during the FIB imaging, deformation of the thin crystal lamella, and variation in the diffraction intensities under electron radiation. These observations provide important information to optimize the FIB milling, and hence is important to obtain high-quality crystal samples for routine structure determination of protein crystals using the electron cryo-microscope.     

Design of spread-out Bragg peaks in hadron therapy with oxygen ions

Aim

Design of a numerical method for creating spread-out Bragg peak (SOBP) and evaluation of the best parameter in Bortfeld Model to this aim in oxygen ion therapy.