An Improved Method for Separating Cardiolipin by HPLC

Herein we report an improved method to separate cardiolipin (Ptd(2)Gro) from tissue total lipid extracts using a biphasic solvent system combined with high performance liquid chromatography. This method uses a normal phase silica column and two mobile phases: mobile phase A that was n-hexane:2-propanol (3:2 by vol) and mobile phase B that was n-hexane:2-propanol:water (56.7:37.8:5.5 by vol). The initial solvent conditions were 95% A and 5% B, with a flow rate of 1.5 mL/min. The samples were from non-derivatized aliquots of liver, heart, or brain lipid extracts. The peak corresponding to Ptd(2)Gro appeared at 31 min, was well defined and did not overlap with neighboring peaks. The adjacent peak corresponded to ethanolamine glycerophospholipids and the remaining phospholipids were eluted in a single peak. The identity of the phospholipids separated by this method was verified by thin layer chromatography (TLC) and fatty acid analysis, which confirmed that the Ptd(2)Gro was well resolved from other phospholipids. This method is useful to separate and quantify Ptd(2)Gro from small tissue samples thereby avoiding the variability associated with TLC methods.     

Stable phase-modulated fiber-optic sensors for senior electricalengineering laboratory curricula

A set of fiber-optic sensing experiments that can be performed by senior undergraduates in a fiber optics laboratory is presented. The experiments are based on two stable phase-modulated sensors: a miniaturized Michelson-type fiber-optic interferometer that is relatively insensitive to temperature drifts; and a two-mode fiber used in a differential interferometric sensing scheme. Methods of construction are described, and several possible experiments are presented. These experiments introduce or strengthen the ideas of interference, elliptical-core fibers, and electromagnetic field propagation in fibers     

Trends in chemical engineering education: Process, product and sustainable chemical engineering challenges

Teaching chemical engineering has always been faced with a dilemma: either keep in touch with industry needs or incorporate new scientific concepts into the curriculum. In this paper, a short historical analysis of the evolution of chemical engineering teaching is presented and the recent trends of the two previous facets (industry and science) are briefly reviewed. The process vs product engineering concept is proposed as one of the means to achieve a better alignment between the curriculum and industry needs. A chemical engineering teaching framework, based in part on a product and a process oriented component, which has been in place in our department 5 years ago, is described and discussed. The concept of sustainable chemistry, including process and product considerations, which can be seen as the next frontier in chemical engineering education, is finally analysed from the education point of view.     

Estradiol Favors the Formation of Eicosapentaenoic Acid (20:5n-3) and n-3 Docosapentaenoic Acid (22:5n-3) from Alpha-Linolenic Acid (18:3n-3) in SH-SY5Y Neuroblastoma Cells

Whether neurosteroids regulate the synthesis of long chain polyunsaturated fatty acids in brain cells is unknown. We examined the influence of 17-β-estradiol (E2) on the capacity of SH-SY5Y cells supplemented with α-linolenic acid (ALA), to produce eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA) and docosahexaenoic acid (DHA). Cells were incubated for 24 or 72 h with ALA added alone or in combination with E2 (ALA + E2). Fatty acids were analyzed by gas chromatography of ethanolamine glycerophospholipids (EtnGpl) and phosphatidylcholine (PtdCho). Incubation for 24 h with ALA alone increased EPA and DPA in EtnGpl, by 330 and 430% compared to controls (P < 0.001) and DHA by only 10% (P < 0.05). Although DHA increased by 30% (P < 0.001) in ALA + E2-treated cells, the difference between the ALA and ALA + E2 treatments were not significant after 24 h (Anova-1, Fisher’s test). After 72 h, EPA, DPA and DHA further increased in EtnGpl and PtdCho of cells supplemented with ALA or ALA + E2. Incubation for 72 h with ALA + E2 specifically increased EPA (+34% in EtnGpl, P < 0.001) and DPA (+15%, P < 0.001) compared to ALA alone. Thus, SH-SY5Y cells produced membrane EPA, DPA and DHA from supplemental ALA. The formation of DHA was limited, even in the presence of E2. E2 significantly favored EPA and DPA production in cells grown for 72 h. Enhanced synthesis of ALA-elongation products in neuroblastoma cells treated with E2 supports the hypothesis that neurosteroids could modulate the metabolism of PUFA.     

Host Recognition by the Specialist Hoverfly <Emphasis Type="Italic">Microdon mutabilis</Emphasis>, a Social Parasite of the Ant <Emphasis Type="Italic">Formica lemani</Emphasis>

The larva of the hoverfly Microdon mutabilis is a specialist social parasite of the ant Formica lemani that is adapted to local groups of F. lemani colonies but mal-adapted to colonies of the same species situated only a few hundred meters away. At a study site in Ireland, F. lemani shares its habitat with four other ant species. All nest under stones, making the oviposition choice by M. mutabilis females crucial to offspring survival. In this study, we tested the hypothesis that, as an extreme specialist, M. mutabilis should respond to cues derived from its host rather than from its microenvironment, a phenomenon that has hitherto only been addressed in the context of herbivorous insects and their parasitoids. In behavioral assays, M. mutabilis females reacted to volatiles from F. lemani colonies by extending their ovipositors, presumably probing for an oviposition substrate. This behavior was not observed toward negative controls or volatiles from colonies of Myrmica scabrinodis, the host ant of the closely related Microdon myrmicae. Coupled gas chromatography-electroantennography (GC-EAG) that used antennal preparations of M. mutabilis located a single physiologically active compound within an extract of heads of F. lemani workers. Coupled GC-mass spectrometry (GC-MS) tentatively identified the compound as a methylated methylsalicylate. GC co-injection of the extract with authentic samples showed that of the four possible isomers (methyl 3-, 4-, 5-, and 6-methylsalicylate), only methyl 6-methylsalicylate co-eluted with the EAG-active peak. Furthermore, the response to methyl 6-methylsalicylate was four times higher than to those of the other isomers. Coupled GC-EAG and GC-MS also revealed physiological responses to two constituents, 3-octanone and 3-octanol, of the M. scabrinodis alarm pheromone. However, the behavioral trials did not reveal any behavior that could be attributed to these compounds. Results are discussed in the context of four phases of host location behavior, and of the characteristics, which volatile cues should provide to be useful for an extreme specialist such as M. mutabilis.     

Synthesis and characterization of cytidine derivatives that inhibit the kinase IspE of the non-mevalonate pathway for isoprenoid biosynthesis

The enzymes of the non-mevalonate pathway for isoprenoid biosynthesis are attractive targets for the development of novel drugs against malaria and tuberculosis. This pathway is used exclusively by the corresponding pathogens, but not by humans. A series of water-soluble, cytidine-based inhibitors that were originally designed for the fourth enzyme in the pathway, IspD, were shown to inhibit the subsequent enzyme, the kinase IspE (from Escherichia coli). The binding mode of the inhibitors was verified by co-crystal structure analysis, using Aquifex aeolicus IspE. The crystal structures represent the first reported example of a co-crystal structure of IspE with a synthetic ligand and confirmed that ligand binding affinity originates mainly from the interactions of the nucleobase moiety in the cytidine binding pocket of the enzyme. In contrast, the appended benzimidazole moieties of the ligands adopt various orientations in the active site and establish only poor intermolecular contacts with the protein. Defined binding sites for sulfate ions and glycerol molecules, components in the crystallization buffer, near the well-conserved ATP-binding Gly-rich loop of IspE were observed. The crystal structures of A. aeolicus IspE nicely complement the one from E. coli IspE for use in structure-based design, namely by providing invaluable structural information for the design of inhibitors targeting IspE from Mycobacterium tuberculosis and Plasmodium falciparum. Similar to the enzymes from these pathogens, A. aeolicus IspE directs the OH group of a tyrosine residue into a pocket in the active site. In the E. coli enzyme, on the other hand, this pocket is lined by phenylalanine and has a more pronounced hydrophobic character.     

Analysis of dynamic plasma behaviours in gas metal arc welding by imaging spectroscopy

For gas metal arc welding, the effect of CO₂ mixture in a shielding gas on a metal transfer process was investigated through the observation of the plasma characteristics and dynamic behaviour at the droplet’s growth-separation-transfer by the temperature measurement methods which were suitable, respectively, to the argon plasma region and the metal plasma region. At the present experimental conditions, the metal transfer process was a spray transfer type with 100%Ar shielding gas. On the other hand, with 85%Ar + 15%CO₂ shielding gas, the metal transfer process was a globular transfer type in which the arc length was shorter, the width was narrower and the time interval of the droplet separation was longer. For both shielding gases, the metal plasma region near the arc central axis exhibited 6500–7500 K, which was lower than the argon plasma region. With 85%Ar + 15%CO₂ shielding gas, when the metal droplet grew below the electrode wire, the region below the droplet has a high plasma temperature and a high concentration of iron vapour which surrounded the droplet. The region also exhibited a remarkably high electron number density. At the spray transfer process, the argon plasma region had an electron number density twice as high as the metal plasma region. Meanwhile, at the globular transfer process, the metal plasma region had a higher electron number density than the argon plasma region, which corresponded to a higher electrical conductivity near the arc axis. This means that the electric current goes through the arc axis easier than the spray transfer process. This condition increases the temperature below the droplet. The thermal expansion increases the force preventing the droplet from falling down. In consequence, the metal transfer takes the globular transfer type.     

Neutrophil Extracellular Traps and Their Implications in Cardiovascular and Inflammatory Disease

Neutrophils are primary effector cells of innate immunity and fight infection by phagocytosis and degranulation. Activated neutrophils also release neutrophil extracellular traps (NETs) in response to a variety of stimuli. These NETs are net-like complexes composed of cell-free DNA, histones and neutrophil granule proteins. Besides the evolutionarily conserved mechanism to capture and eliminate pathogens, NETs are also associated with pathophysiological processes of various diseases. Here, we elucidate the mechanisms of NET formation and their different implications in disease. We focused on autoinflammatory and cardiovascular disorders as the leading cause of death. Neutrophil extracellular traps are not only present in various cardiovascular diseases but play an essential role in atherosclerotic plaque formation, arterial and venous thrombosis, as well as in the development and progression of abdominal aortic aneurysms. Furthermore, NETosis can be considered as a source of autoantigens and maintains an inflammatory milieu promoting autoimmune diseases. Indeed, there is further need for research into the balance between NET induction, inhibition, and degradation in order to pharmacologically target NETs and their compounds without impairing the patient’s immune defense. This review may be of interest to both basic scientists and clinicians to stimulate translational research and innovative clinical approaches.     

High-Pressure Reverse Osmosis for Industrial Water Recycling: Permeate-Sided Pressure Drop as Performance-Limiting Factor

In the chemical industry large amounts of saline wastewater occur. Its disposal into rivers is a considerable burden to the ecosystem. To strive for a circular economy and enable a viable raw material recycling, energy-efficient concentration processes are requisite. High-pressure reverse osmosis meets this criterion, but its industrial application demands suitable membrane elements that withstand the exceptional operation conditions and provide sufficient performance. Hence, new requirements regarding the design of spiral-wound elements arise. To identify those, specific performance-limiting effects need a better understanding.     

Idiosyncratic Drug-Induced Liver Injury: Mechanistic and Clinical Challenges

Idiosyncratic drug-induced liver injury (IDILI) remains a significant problem for patients and drug development. The idiosyncratic nature of IDILI makes mechanistic studies difficult, and little is known of its pathogenesis for certain. Circumstantial evidence suggests that most, but not all, IDILI is caused by reactive metabolites of drugs that are bioactivated by cytochromes P450 and other enzymes in the liver. Additionally, there is overwhelming evidence that most IDILI is mediated by the adaptive immune system; one example being the association of IDILI caused by specific drugs with specific human leukocyte antigen (HLA) haplotypes, and this may in part explain the idiosyncratic nature of these reactions. The T cell receptor repertoire likely also contributes to the idiosyncratic nature. Although most of the liver injury is likely mediated by the adaptive immune system, specifically cytotoxic CD8+ T cells, adaptive immune activation first requires an innate immune response to activate antigen presenting cells and produce cytokines required for T cell proliferation. This innate response is likely caused by either a reactive metabolite or some form of cell stress that is clinically silent but not idiosyncratic. If this is true it would make it possible to study the early steps in the immune response that in some patients can lead to IDILI. Other hypotheses have been proposed, such as mitochondrial injury, inhibition of the bile salt export pump, unfolded protein response, and oxidative stress although, in most cases, it is likely that they are also involved in the initiation of an immune response rather than representing a completely separate mechanism. Using the clinical manifestations of liver injury from a number of examples of IDILI-associated drugs, this review aims to summarize and illustrate these mechanistic hypotheses.