No evidence for radiation-induced clastogenic factors after in vitro or in vivo exposure of human blood

Experiments were performed with human plasma irradiated in vitro or in vivo in order to evaluate the extent to which clastogenic factors might disturb the adaptive response to DNA-damaging factors currently studied in our laboratory. The studies were carried out with plasma isolated from whole blood given 4 Gy of X-rays in vitro and with plasma from people receiving local radiotherapy at a total dose of about 60 Gy gamma rays. Addition of irradiated plasma to culture medium did not result in a statistically significant increase in structural aberrations in chromosomes of non-irradiated normal blood.     

Photobiological and Structural Studies of Light-driven Movements in the Solar-tracking Leaf of Lupinus palaestinus Bioss. (Fabaceae)

The compound, palmate lamina of Lupinus palaestinus reorients photonastically, as well as phototropically in response to non-directional and directional light signals, respectively, by structural deformations of pulvini. When the excitation provided by directional light is maintained constant (fluence rate, angle of incidence and azimuth, with respect to the leaflet laminae), the entire lamina reorients towards it at a constant angular velocity over a considerable time interval and displacement. The laminar pulvinules are considerably longer than the subtending common petiolar pulvinus and therefore contribute most to laminar reorientation. The pulvinar region is characterized by transverse folds around its circumference, and longitudinal rib-like thickenings on the external walls of its epidermis that facilitate axial and transverse deformations. Specialized “joints”, at the distal and proximal ends of each pulvinule, contribute most to its flexing. Anthocyanin is notable by its absence. Specialized “motor” tissues surrounding the central vascular core participate in pulvinar deformation by undergoing directional and differential volume changes. The bundle sheath is characterized by numerous starch grains. The multi-layered cortical parenchyma exhibits an abundance of transversely oriented primary pit fields and associated plasmodesmata. When the leaflet lamina rotates around its midrib, the pulvinus twists along its axis, exhibiting epidermal and cortical deformation. The functional significance of these specializations is discussed.     

Performance of the marginal structural models under various scenarios of incomplete marker's values: A simulation study

Marginal structural models (MSMs) have been proposed for estimating a treatment's effect, in the presence of time‐dependent confounding. We aimed to evaluate the performance of the Cox MSM in the presence of missing data and to explore methods to adjust for missingness. We simulated data with a continuous time‐dependent confounder and a binary treatment. We explored two classes of missing data: (i) missed visits, which resemble clinical cohort studies; (ii) missing confounder's values, which correspond to interval cohort studies. Missing data were generated under various mechanisms. In the first class, the source of the bias was the extreme treatment weights. Truncation or normalization improved estimation. Therefore, particular attention must be paid to the distribution of weights, and truncation or normalization should be applied if extreme weights are noticed. In the second case, bias was due to the misspecification of the treatment model. Last observation carried forward (LOCF), multiple imputation (MI), and inverse probability of missingness weighting (IPMW) were used to correct for the missingness. We found that alternatives, especially the IPMW method, perform better than the classic LOCF method. Nevertheless, in situations with high marker's variance and rarely recorded measurements none of the examined method adequately corrected the bias.     

Structure Basis for Shaping the Nse4 Protein by the Nse1 and Nse3 Dimer within the Smc5/6 Complex

The Smc5/6 complex facilitates chromosome replication and DNA break repair. Within this complex, a subcomplex composed of Nse1, Nse3 and Nse4 is thought to play multiple roles through DNA binding and regulating ATP-dependent activities of the complex. However, how the Nse1-Nse3-Nse4 subcomplex carries out these multiple functions remain unclear. To address this question, we determine the crystal structure of the Xenopus laevis Nse1-Nse3-Nse4 subcomplex at 1.7 Å resolution and examine how it interacts with DNA. Our structural analyses show that the Nse1-Nse3 dimer adopts a closed conformation and forms three interfaces with a segment of Nse4, forcing it into a Z-shaped conformation. The Nse1-Nse3-Nse4 structure provides an explanation for how the lung disease immunodeficiency and chromosome breakage syndrome-causing mutations could dislodge Nse4 from Nse1-Nse3. Our DNA binding and mutational analyses reveal that the N-terminal and the middle region of Nse4 contribute to DNA interaction and cell viability. Integrating our data with previous crosslink mass spectrometry data, we propose potential roles of the Nse1-Nse3-Nse4 complex in binding DNA within the Smc5/6 complex.     

Structural basis for IL-12 and IL-23 receptor sharing reveals a gateway for shaping actions on T versus NK cells

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Preparation and Properties of Human Kappa-casein-like Fraction

A κ-casein-like fraction was prepared from human whole casein by gel filtration with Sephadex G-150 and Biogel A-150 m. The fraction was calcium-insensitive and its solution became turbid by rennin. In polyacrylamide gel electrophoretic (PAE) analysis, the fraction gave 11~12 bands after reduction with 2-mercaptoethanol. It appeared to exist as a disulfide- bound complex of many components. The occurrence of human para-IC-caseins from the fraction after rennin treatment was confirmed by PAE. When the reduced, alkylated human κ-casein-like fraction was chromatographed by DEAE-cellulose, several fractions were obtained. After rennin treatment, they formed either a para-SCM-κ-casein band moving toward the cathode on PAE pattern or the one which moved toward the anode. These results suggest that two para-κ-caseins were formed from human whole casein or human κ-casein-like fraction by the action of rennin.     

Module-based construction of plasmids for chromosomal integration of the fission yeast Schizosaccharomyces pombe

Integration of an external gene into a fission yeast chromosome is useful to investigate the effect of the gene product. An easy way to knock-in a gene construct is use of an integration plasmid, which can be targeted and inserted to a chromosome through homologous recombination. Despite the advantage of integration, construction of integration plasmids is energy- and time-consuming, because there is no systematic library of integration plasmids with various promoters, fluorescent protein tags, terminators and selection markers; therefore, researchers are often forced to make appropriate ones through multiple rounds of cloning procedures. Here, we establish materials and methods to easily construct integration plasmids. We introduce a convenient cloning system based on Golden Gate DNA shuffling, which enables the connection of multiple DNA fragments at once: any kind of promoters and terminators, the gene of interest, in combination with any fluorescent protein tag genes and any selection markers. Each of those DNA fragments, called a ‘module’, can be tandemly ligated in the order we desire in a single reaction, which yields a circular plasmid in a one-step manner. The resulting plasmids can be integrated through standard methods for transformation. Thus, these materials and methods help easy construction of knock-in strains, and this will further increase the value of fission yeast as a model organism.     

墨江蜈蚣与少棘蜈蚣的比较研究──Ⅱ．药效和毒理

本文比较了墨江蜈蚣与少棘蜈蚣的药效学和毒理学作用，实验包括抗惊厥试验、对致病性真菌和细菌体外生长的影响、急性毒性试验和染色体畸变实验等。结果两种蜈蚣的３％醋酸提取液对所试的真菌有抑制作用但无杀真菌作用，而对细菌无抑制作用。两种蜈蚣的毒性很低，在给小鼠５０ｇ／ｋｇ的剂量下都无法测出ＬＤ５０。在２０５ｍｇ／ｋｇ条件下的致突变率与未给药的突变率接近，表明两者都无遗传毒性。