Molecular dynamics simulation of single-walled carbon nanotubes inside liquid crystals

Molecular dynamic simulations of systems of single-walled carbon nanotubes (CNTs) in liquid crystalline solvents were performed, in order to investigate the effect of the molecular structure and phase of the liquid crystal (LC) on the interactions between the CNTs. Three different LC molecules (5CB, 8CB and 5CF) were considered in our study. Our results with 5CB and 8CB suggest that increasing the chain length of the hydrophobic part of the LC molecule by three carbon atoms is insufficient to decrease the tendency for the CNTs to aggregate in the LCs. Additionally, varying the phase of the LC is also insufficient to decrease the aggregation tendency of the CNTs. However, simulations with 5CF (which has fluorine atoms in the head group of the LC molecule) suggest that this LC solvent can decrease the tendency of the CNTs to aggregate. This study is relevant to assist experimentalists with the development of high-quality dispersions of large concentrations of CNTs in the LCs.     

Inositol 1,4,5-trisphosphate receptor-isoform diversity in cell death and survival

Cell-death and -survival decisions are critically controlled by intracellular Ca^2 + homeostasis and dynamics at the level of the endoplasmic reticulum (ER). Inositol 1,4,5-trisphosphate (IP₃) receptors (IP₃Rs) play a pivotal role in these processes by mediating Ca^2 + flux from the ER into the cytosol and mitochondria. Hence, it is clear that many pro-survival and pro-death signaling pathways and proteins affect Ca^2 + signaling by directly targeting IP₃R channels, which can happen in an IP₃R-isoform-dependent manner. In this review, we will focus on how the different IP₃R isoforms (IP₃R1, IP₃R2 and IP₃R3) control cell death and survival. First, we will present an overview of the isoform-specific regulation of IP₃Rs by cellular factors like IP₃, Ca^2 +, Ca^2 +-binding proteins, adenosine triphosphate (ATP), thiol modification, phosphorylation and interacting proteins, and of IP₃R-isoform specific expression patterns. Second, we will discuss the role of the ER as a Ca^2 + store in cell death and survival and how IP₃Rs and pro-survival/pro-death proteins can modulate the basal ER Ca^2 + leak. Third, we will review the regulation of the Ca^2 +-flux properties of the IP₃R isoforms by the ER-resident and by the cytoplasmic proteins involved in cell death and survival as well as by redox regulation. Hence, we aim to highlight the specific roles of the various IP₃R isoforms in cell-death and -survival signaling. This article is part of a Special Issue entitled: Calcium signaling in health and disease. Guest Editors: Geert Bultynck, Jacques Haiech, Claus W. Heizmann, Joachim Krebs, and Marc Moreau.     

Hemoglobin regulates the migration of glioma cells along poly(ε‐caprolactone)‐aligned nanofibers

Aligned fibers have been shown to facilitate cell migration in the direction of fiber alignment while oxygen (O₂)‐carrying solutions improve the metabolism of cells in hypoxic culture. Therefore, U251 aggregate migration on poly(ε‐caprolactone) (PCL)‐aligned fibers was studied in cell culture media supplemented with the O₂ storage and transport protein hemoglobin (Hb) obtained from bovine, earthworm and human sources at concentrations ranging from 0 to 5 g/L within a cell culture incubator exposed to O₂ tensions ranging from 1 to 19% O₂. Individual cell migration was quantified using a wound healing assay. In addition, U251 cell aggregates were developed and aggregate dispersion/cell migration quantified on PCL‐aligned fibers. The results of this work show that the presence of bovine or earthworm Hb improved individual cell viability at 1% O₂, while human Hb adversely affected cell viability at increasing Hb concentrations and decreasing O₂ levels. The control data suggests that decreasing the O₂ tension in the incubator from 5 to 1% O₂ decreased aggregate dispersion on the PCL‐aligned fibers. However, the addition of bovine Hb at 5% O₂ significantly improved aggregate dispersion. At 19% O₂, Hb did not impact aggregate dispersion. Also at 1% O₂, aggregate dispersion appeared to increase in the presence of earthworm Hb, but only at the latter time points. Taken together, these results show that Hb‐based O₂ carriers can be utilized to improve O₂ availability and the migration of glioma spheroids on nanofibers. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:1214–1220, 2014     

Familial secondary erythrocytosis due to increased oxygen affinity is caused by destabilization of the T state of hemoglobin Brigham (α(2) β(2) (Pro100Leu) )

Hemoglobin Brigham (β Pro100 to Leu) was first reported in a patient with familial erythrocytosis. Erythrocytes of an affected individual from the same family contain both HbA and Hb Brigham and exhibit elevated O(2) affinity compared with normal cells (P(50) = 23 mm Hg vs. 31 mmHg at pH 7.4 at 37°C). O(2) affinities measured for hemolysates were sensitive to changes in pH or chloride concentrations, indicating little change in the Bohr and Chloride effects. Hb Brigham was separated from normal HbA by nondenaturing cation exchange liquid chromatography, and the amino acid substitution was verified by mass spectrometry. The properties of Hb Brigham isolated from the patient's blood were then compared with those of recombinant Hb Brigham expressed in Escherichia coli. Kinetic experiments suggest that the rate constants for ligand binding and release in the high (R) and low (T) affinity quaternary states of Hb Brigham are similar to those of native hemoglobin. However, the Brigham mutation decreases the T to R equilibrium constant (L) which accelerates the switch to the R state during ligand binding to deoxy-Hb, increasing the rate of association by approximately twofold, and decelerates the switch during ligand dissociation from HbO(2) , decreasing the rate approximately twofold. These kinetic data help explain the high O(2) affinity characteristics of Hb Brigham and provide further evidence for the importance of the contribution of Pro100 to intersubunit contacts and stabilization of the T quaternary structure.     

Action mechanism of tachyplesin I and effects of PEGylation

PEGylation of protein and peptide drugs is frequently used to improve in vivo efficacy. We investigated the action mechanism of tachyplesin I, a membrane-acting cyclic antimicrobial peptide from Tachypleus tridentatus and the effects of PEGylation on the mechanism. The PEGylated peptide induced the leakage of calcein from egg yolk l-α-phosphatidylglycerol/egg yolk l-α-phosphatidylcholine large unilamellar vesicles similarly to the parent peptide. Both peptides induced lipid flip-flop coupled to leakage and was translocated into the inner leaflet of the bilayer, indicating that tachyplesin I forms a toroidal pore and that PEGylation did not alter the basic mechanism of membrane permeabilization of the parent peptide. Despite their similar activities against model membranes, the peptides showed very different biological activities. The cytotoxicity of tachyplesin I was greatly reduced by PEGylation, although the antimicrobial activity was significantly weakened. We investigated the enhancement of the permeability of inner membranes induced by the peptides. Our results suggested that outer membranes and peptidoglycan layers play an inhibitory role in the permeation of the PEG moiety. Furthermore, a reduction in DNA binding by PEGylation may also contribute to the weak activity of the PEGylated peptide.     

液体培养条件下细菌浓度两种测定方法比较

液体培养细菌后,分别测定培养液、培养液沉淀 无菌盐水的吸光度,结果存在明显差异。采用稀释平板法和显微计数法测定了培养液中细菌数量,并推导出嗜水气单胞菌活菌数与吸光度的关系为logI0/I=6.848×10-25·C。     

A globin gene of ancient evolutionary origin in lower vertebrates: evidence for two distinct globin families in animals

Hemoglobin, myoglobin, neuroglobin, and cytoglobin are four types of vertebrate globins with distinct tissue distributions and functions. Here, we report the identification of a fifth and novel globin gene from fish and amphibians, which has apparently been lost in the evolution of higher vertebrates (Amniota). Because its function is presently unknown, we tentatively call it globin X (GbX). Globin X sequences were obtained from three fish species, the zebrafish Danio rerio, the goldfish Carassius auratus, and the pufferfish Tetraodon nigroviridis, and the clawed frog Silurana tropicalis. Globin X sequences are distinct from vertebrate hemoglobins, myoglobins, neuroglobins, and cytoglobins. Globin X displays the highest identity scores with neuroglobin (approximately 26% to 35%), although it is not a neuronal protein, as revealed by RT-PCR experiments on goldfish RNA from various tissues. The distal ligand-binding and the proximal heme-binding histidines (E7 and F8), as well as the conserved phenylalanine CD1 are present in the globin X sequences, but because of extensions at the N-terminal and C-terminal, the globin X proteins are longer than the typical eight alpha-helical globins and comprise about 200 amino acids. In addition to the conserved globin introns at helix positions B12.2 and G7.0, the globin X genes contain two introns in E10.2 and H10.0. The intron in E10.2 is shifted by 1 bp in respect to the vertebrate neuroglobin gene (E11.0), providing possible evidence for an intron sliding event. Phylogenetic analyses confirm an ancient evolutionary relationship of globin X with neuroglobin and suggest the existence of two distinct globin types in the last common ancestor of Protostomia and Deuterostomia.     

Mutational study of the bacterial hemoglobin distal heme pocket

Ligand binding experiments on three mutants in the distal heme pocket of Vitreoscilla hemoglobin (GlnE7His, ProE8Ala, and GlnE7His,ProE8Ala) were used to probe the role of GlnE7 and ProE8 in the pocket's unusual structure. The oxygen dissociation constants for the wild type, E8Ala mutant, and E7His mutant proteins were 4.5, 4.7, and 1.7microM, respectively; the K(d) for the double mutant was not determinable by our technique. Visible-Soret spectra of the carbonyl and cyanyl forms and FT-IR of the carbonyl form of the E8 mutant were similar to those of the wild type; the opposite was true for the GlnE7His and GlnE7His,ProE8Ala mutants, which also differed from wild type in the visible-Soret spectra of their oxidized forms. Models of the effects of the mutations on distal pocket structure were consistent with the experimental findings, particularly the larger effects of the GlnE7His change.     

Effect of soil moisture on leaf ecophysiology of <Emphasis Type="Italic">Parasenecio yatabei</Emphasis>, a summer-green herb in a cool–temperate forest understory in Japan

Leaf physiological and gas-exchange traits of a summer-green herbaceous perennial, Parasenecio yatabei, growing along a stream were examined in relation to leaf age. In its vegetative phase, the aerial part of this plant consists of only one leaf and provides an ideal system for the study of leaf longevity. Volumetric soil water content (SWC) decreased with increasing distance from the stream, whereas relative light intensity was nearly constant. The light-saturated net CO₂ assimilation rate (A _sat) and leaf stomatal conductance (gs) were approximately 1.5-fold and 1.4-fold higher, respectively, in the lower slope near the mountain stream than in the upper slope far from the mountain stream. The lifespan of aerial parts of vegetative plants significantly increased with decreasing SWC. The leaf mass-based nitrogen content of the leaves (N _mass) was almost constant (ca. 2.2%); however, the maximum carboxylation rate by ribulose-1,5-biphosphate carboxylase/oxygenase (rubisco) (V _cmax) and photosynthetic nitrogen use efficiency (PNUE, A _sat/N _area) decreased more slowly in the upper slope than in the lower slope. The higher leaf photosynthetic activity of P. yatabei plants growing lower on the slope leads to a decrease in V _cmax and PNUE in the early growing season, and to a shorter leaf lifespan.     

青霉素钠对萼花臂尾轮虫适合度的促进作用及其与藻密度的关系

以萼花臂尾轮虫（Brachionus calyciflorus）为受试动物,在1.0×10⁶（较低）、2.0×10⁶（中等）和4.0×10⁶个细胞/mL （较高）的斜生栅藻（Scenedesmus obliquus）密度下,研究了不同浓度（0、200、400、600、800和1000 μg/L）的青霉素钠对轮虫各生命表统计学参数的影响。结果表明,与各藻密度下的对照组相比,当藻密度为2.0×10⁶个细胞/mL时,200-1000 μg/L的青霉素钠处理组中轮虫的生命期望和世代时间分别延长了40.67%-70.67%和34.04%-50.23%（P<0.05）,净生殖率和内禀增长率分别提高了204.35%-358.70%和36.26%-62.09%（P<0.05）;藻密度的降低或升高均显著降低青霉素钠对轮虫存活、生殖和种群增长的促进幅度。1.0×10⁶ 个细胞/mL的斜生栅藻密度下,400 μg/L的青霉素钠处理组中轮虫的生命期望延长了21%（P<0.05）,200-400μg/L的青霉素钠处理组中轮虫的世代时间延长了23.15%-33.13%（P<0.05）,200-600和1000 μg/L的青霉素钠处理组中轮虫的净生殖率分别升高了40%-81.05%和41.05%（P<0.05）;但4.0×10⁶个细胞/mL的斜生栅藻密度下,各青霉素钠处理组中轮虫的所有生命表统计学参数均与对照组之间无显著性差异（P>0.05）。藻密度对轮虫的生命期望、净生殖率、内禀增长率和后代混交率均具有显著性影响（P<0.01）,青霉素钠浓度对轮虫的世代时间、净生殖率和内禀增长率均具有显著性影响（P<0.05）,藻密度和青霉素钠浓度的交互作用对轮虫的生命期望、净生殖率和内禀增长率也具有显著性影响（P<0.05）。在实验设置的青霉素钠浓度范围内,1.0×10⁶个细胞/mL藻密度下轮虫的生命期望、世代时间和净生殖率与青霉素钠浓度之间均具有显著的剂量-效应关系（P<0.05）;2.0×10⁶个细胞/mL藻密度下,轮虫的生命期望、世代时间和内禀增长率与青霉素钠浓度之间均具有显著的剂量-效应关系（P<0.05）。本研究结果提示,环境相关浓度的青霉素钠不会对萼花臂尾轮虫的存活、生殖和种群增长产生显著性影响。