Coordinate packaging of newly synthesized phosphatidylcholine and phosphatidylglycerol in lamellar bodies in alveolar type II cells

Methylamine, a weak base, inhibits packaging of newly synthesized phosphatidylcholine (PC) in lamellar bodies in 20–22 h cultured alveolar type II cells, suggesting a role for acidic pH of lamellar bodies. In this study, we tested if (i) the packaging of PC is similarly regulated in freshly isolated type II cells and (ii) methylamine also inhibits the packaging of other surfactant phospholipids, particularly, phosphatidylglycerol (PG). The latter would suggest coordinated packaging so as to maintain the phospholipid composition of lung surfactant. During the short-term metabolic labeling experiments in freshly isolated type II cells, methylamine treatment decreased the incorporation of radioactive precursors into PC, disaturated PC (DSPC), and PG of lamellar bodies but not of the microsomes, when compared with controls. The calculated packaging (the percentage of microsomal lipid packaged in lamellar bodies) of each phospholipid was similarly decreased (∼50%) in methylamine-treated cells, suggesting coordinated packaging of surfactant phospholipids in lamellar bodies. Equilibrium-labeling studies with freshly isolated type II cells (as is routinely done for studies on surfactant secretion) ± methylamine showed that in methylamine-treated cells, the secretion of PC and PG was decreased (possibly due to decreased packaging), but the phospholipid composition of released surfactant (measured by radioactivity distribution) was unchanged; and the PC content (measured by mass or radioactivity) of lamellar bodies was lower, but the PC composition (as percentage of total phospholipids) was unchanged when compared with control cells. We speculate that the newly synthesized surfactant phospholipids, PC, DSPC, and PG, are coordinately transported into lamellar bodies by a mechanism requiring the acidic pH, presumably, of lamellar bodies.     

Erythrocyte membrane lipid alterations in undernourished cerebral palsied children during high intakes of a soy oil-based enteral formula

Five undernourished children with severe cerebral palsy (CP) were tube-fed sufficient volumes of Isocal^TM to allow rapid weight gain. Isocal^TM provided, on average, 88% of their daily energy intake for at least 25 days. The purpose of our study was to correct the undernutrition and to analyze the major erythrocyte phospholipids before and after feeding periods for possible feeding and disease-related differences. The fatty acid profiles of erythrocyte membranes from CP children were compared with those from 12 healthy children and with the fatty acid composition of the formula. There were no clinical or biochemical indications of essential fatty acid deficiency. The feeding of a soy oil-based formula increased the proportions of 18∶2n−6 in the phospholipids. The increases occurred predominantly in phosphatidylcholine followed by phosphatidylethanolamine. Despite such large dietary intakes of soy oil, no changes were observed in the phospholipid concentrations of 20∶4n−6, 18∶3n−3, 20∶5n−3, or in the C₂₂n−6 and C₂₂n−3 fatty acids. These findings are consistent with an inhibition of the Δ6 desaturase by high dietary linoleate.     

Distribution of the phosphatidate phosphohydrolase activity in the lamellar body and lysosomal fractions of lung tissue

Phosphatidate phosphohydrolase (PAPase) activity was measured in lamellar bodies purified from porcine lung tissue. After repeated freeze-thawing, only a negligible amount of PAPase activity was released into the soluble fractions, whereas there was release of 2 lysosomal marker enzyme activities, glucosaminidase and β-galactosidase into the soluble fraction. In addition, a lysosomal-enriched fraction was prepared from adult rat lung tissue by prior treatment of the rats with Triton WR 1339. Treatment with Triton WR 1339 resulted in the significant shift of the activities of the lysosomal marker enzymes, glucosaminidase and β-galactosidase, to less dense subcellular fractions. The highest specific activity of PAPase was found in a subcellular fraction which had a density that was intermediate between that of the mitochondrial and microsomal fractions and the distribution of PAPase activity was not affected by the prior treatment of the rats with Triton WR 1339.     

Insulin status-dependent alterations in lipid/phospholipid composition of rat kidney microsomes and mitochondria

Early and late effects of alloxan-diabetes on lipid/phospholipid composition in rat kidney microsomes and mitochondria were examined. In microsomes, early diabetic state resulted in an increase in contents of total phospholipids (TPL), cholesterol (CHL), with an increase in the lysophospholipids (Lyso), phosphatidylcholine (PC), and phosphatidylinositol (PI) components. The sphinogmyelin (SPM), phosphatidylethanolamine (PE), phosphatidylserine (PS), and phosphatidic acid (PA) content decreased. Treatment with insulin had no effect on PC but PE increased and the other components decreased. In the 1-month diabetic group PI, PS, PE, and PA components decreased, whereas Lyso and PC increased. Treatment with insulin had restorative effects on PE, PI, and PS; Lyso was further elevated whereas PA decreased. In mitochondria, at an early stage of diabetes marginally increased CHL content was restored by insulin treatment. Long-term diabetes lowered the TPL and elevated the CHL content. Treatment with insulin partially restored the TPL and CHL content. A diabetic state decreased the proportion of PE and diphosphatidylglycerol (DPG) components but increased the Lyso, SPM, PC, PI, and PS components in the mitochondria. Treatment with insulin had a partial restorative effect. The membrane fluidity of both microsomes and mitochondria decreased in general in the diabetic condition and was not corrected by insulin treatment at a late stage. However, at an early stage, treatment with insulin fluidized both membranes.     

Essential metalloelement chelates facilitate repair of radiation injury

Treatment with essential metalloelement (Cu, Fe, Mn, and Zn) chelates or combinations of them before and/or after radiation injury is a useful approach to overcoming radiation injury. No other agents are known to increase survival when they are used to treat after irradiation, in a radiorecovery treatment paradigm. These chelates may be useful in facilitating de novo syntheses of essential metalloelement-dependent enzymes required to repair radiation injury. Reports of radioprotection, which involves treatment before irradiation, with calcium-channel blockers, acyl Melatonin homologs, and substituted anilines, which may serve as chelating agents after biochemical modification in vivo, as well as Curcumin, which is a chelating agent, have been included in this review. These inclusions are intended to suggest additional approaches to combination treatments that may be useful in facilitating radiation recovery. These approaches to radioprotection and radiorecovery offer promise in facilitating recovery from radiation-induced injury experienced by patients undergoing radiotherapy for neoplastic disease and by individuals who experience environmental, occupational, or accidental exposure to ultraviolet, x-ray, or gamma-ray radiation. Since there are no existing treatments of radiation-injury intended to facilitate tissue repair, studies of essential metalloelement chelates and combinations of them, as well as combinations of them with existing organic radioprotectants, seem worthwhile.     

Effect of inhibitors and phenobarbital pretreatment upon hepatic lipid peroxidation during protein and riboflavin dietary stress in male rats

Male young albino rats divided into three groups were maintained on the following diets. The normal group was maintained on 13% casein, 45% corn starch, 31% sucrose, 6% salt mixture, 4% peanut oil, and 1% vitamin mixture. The low protein group animals recieved only 5% casein, and the riboflavin-deficient group was fed normal diet, except that the riboflavin was absent from the vitamin mixture and ordinary casein was replaced by 13% vitamin-free casein. The effects of various inhibitors upon triphosphopyridine nucleotide, reduced form-linked lipid peroxide formation by the supernatant fraction of liver at 9000 × g from rats fed a normal diet, a low protein diet, or a riboflavin-deficient diet for 2,4, and 7 weeks were investigated. A significant decrease in triphosphoryridine nucleotide, reduced form-linked and ascorbate-induced lipid peroxidation was noticed in rats fed on low protein and riboflavin-deficient diet. Glutathione inhibited the triphosphopyridine nucleotide, reduced form-linked lipid peroxidation in rats from all three groups. However, the observed response was variable due to the nature of the diet. The magnitude of inhibition was greater in low protein-fed animals than in animals from the riboflavin-deficient and control groups. Cytochrome C inhibited peroxide formation, but the inhibition was greater in rats from the low protein and riboflavin-deficient groups than in animals from the normal group. Tocopherol exhibited the antioxidant property in all three groups of rats. Deoxycholate inhibited lipid peroxide formation in all the three groups.     

直升机机体“鼓包”损伤的胶接修理

针对某型直升机机体“鼓包”损伤,分析了损伤的原因,采用不同的胶接修理方法进行修理,达到了较好的效果。     

Fluorescent products of phospholipids during lipid peroxidation

During peroxidation, phosphatidyl ethanolamine and phosphatidyl serine formed fluorescent chromophores with maximum emission at 435 nm and maximum excitation at 365 nm. The development of fluorescence was related to formation of thiobarbituric acid reactive substance during lipid peroxidation. This relationship was studied by reacting dipalmityl phosphatidyl ethanolamine with the oxidation products of the methyl esters of arachidonic, linolenic and linoleic fatty acids. Reaction parameters affecting the development of lipid-extractable fluorescent chromophores are: the production of peroxidation products, especially malonaldehyde, from autoxidation of polyunsaturated fatty acids; the length of time these products react; and the availability of reactive amino groups on the phospholipids.     

Bioorthogonal chemical reporters for analyzing protein lipidation and lipid trafficking

Protein lipidation and lipid trafficking control many key biological functions in all kingdoms of life. The discovery of diverse lipid species and their covalent attachment to many proteins has revealed a complex and regulated network of membranes and lipidated proteins that are central to fundamental aspects of physiology and human disease. Given the complexity of lipid trafficking and the protein targeting mechanisms involved with membrane lipids, precise and sensitive methods are needed to monitor and identify these hydrophobic molecules in bacteria, yeast, and higher eukaryotes. Although many analytical methods have been developed for characterizing membrane lipids and covalently modified proteins, traditional reagents and approaches have limited sensitivity, do not faithfully report on the lipids of interest, or are not readily accessible. The invention of bioorthogonal ligation reactions, such as the Staudinger ligation and azide-alkyne cycloadditions, has provided new tools to address these limitations, and their use has begun to yield fresh insight into the biology of protein lipidation and lipid trafficking. In this Account, we discuss how these new bioorthogonal ligation reactions and lipid chemical reporters afford new opportunities for exploring the biology of lipid-modified proteins and lipid trafficking. Lipid chemical reporters from our laboratory and several other research groups have enabled improved detection and large-scale proteomic analysis of fatty-acylated and prenylated proteins. For example, fatty acid and isoprenoid chemical reporters in conjunction with bioorthogonal ligation methods have circumvented the limited sensitivity and hazards of radioactive analogues, allowing rapid and robust fluorescent detection of lipidated proteins in all organisms tested. These chemical tools have revealed alterations in protein lipidation in different cellular states and are beginning to provide unique insights in mechanisms of regulation. Notably, the purification of proteins labeled with lipid chemical reporters has allowed both the large-scale analysis of lipidated proteins as well as the discovery of new lipidated proteins involved in metabolism, gene expression, and innate immunity. Specific lipid reporters have also been developed to monitor the trafficking of soluble lipids; these species are enabling bioorthogonal imaging of membranes in cells and tissues. Future advances in bioorthogonal chemistry, specific lipid reporters, and spectroscopy should provide important new insight into the functional roles of lipidated proteins and membranes in biology.     

Morphology of undeformed and deformed polyethylene lamellar crystals

Morphology of undeformed polyethylene crystals obtained by high pressure crystallization was investigated by SEM. It was revealed by exposing the interior of the samples by microtoming followed by permanganic etching. The etching procedure was refined to reveal defected sites of lamellae in addition to differentiation of crystalline and amorphous phases. From 1 to 3 screw dislocations with large Burgers vector per 1 μm² of lamellae basal planes were detected. Lamellae, when viewed edge-on give an impression of a “blocky architecture”, while their real shape, as seen on SEM images of flat-on and oblique lamellae, resembles platelets with a few defects in the form of screw dislocations protruding a platelet.High pressure-crystallized polyethylene samples were deformed plastically by uniaxial compression and were studied by SEM and AFM. When the deformation is interrupted dislocations are arrested within the crystals. It was observed that in contrast to undeformed samples, the side faces of deformed lamellae were not any longer smooth and a large number of screw dislocations with low Burgers vectors crossing the lamellae thickness could be distinguished. These observations are in accordance with polymer crystal plasticity theory that relies on the rate controlled nucleation and propagation of screw dislocations across polymer crystals. An existence of numerous screw dislocations arrested in lamellae is a direct proof of action of fine crystallographic slips along the macromolecular chains in PE crystals during plastic deformation. The kinking of lamellae due to plastic deformation was also observed. Large sections of lamellae between kinks rotated towards the plane of compression while the chain stems in lamellae rotated in the opposite direction, away from the compression direction, which is a signature of the fine crystallographic slip.Plastically deformed polyethylene crystals are highly defected due to many dislocations incorporated within them - the density of dislocations was approximated as 10¹⁶ m⁻². However, deformed crystal melting temperature is nearly unaffected while the heat of melting is slightly reduced, yet only in thin crystals. It suggests that the arrested dislocations contribute more to the surface energy of lamellae basal planes rather than to a bulk energy of polyethylene crystals.     

球罐法兰焊缝损坏的特殊修复方法

多晶硅生产工艺中,盛装氯硅烷球罐本体上的法兰焊缝损坏,在无法将设备内部的氯硅烷彻底置换干净的情况下,应用简易制作的活塞杆塞进法兰短接处,起到密封作用,在保证洁净度的前提下,使设备内部与需修复的法兰焊缝处完全隔离,通过动火分析合格后,对损坏的焊缝进行修复,确保了设备的安全运行,同时,对以后处理相类似的问题提供了借鉴。     

Structure and properties of graphite-chromyl chloride lamellar compound

Crystal structure, electric conductivity and thermal stability of graphite-CrO₂Cl₂ lamellar compound were investigated. X-ray diffraction studies of C_27·5CrO₂Cl₂ show that it possesses a 3rd stage lamellar structure with I_c of 14·87 Å. This lamellar compound is unusually stable and its electric conductivity, as well as its composition, show only a slight change after complete evacuation of reacting CrO₂Cl₂ gas. When heattreated in a vacuum, its electric conductivity shows a considerable decrease, whereas its composition changes very slowly up to 300°C.     

Self-assembly template during morphological transition of a linear ABC triblock copolymer from lamellar to Gyroid structure

Morphological transition of the ABC-type triblock copolymer from lameller to tricontinuous Gyroid structure has been observed. The sample used is a symmetric poly(isoprene-block-styrene-block-2-vinylpyridine), whose volume fractions of three components are 0.26, 0.48 and 0.26, and the total molecular weight is 1.0×10⁵. The solvent cast films were prepared and dried well, followed by annealing at 423 K, the temperature well above the glass transition temperatures of three polymer components. The structural transition was investigated by varying the annealing time from 1 h up to 7 days with two methods, transmission electron microscopy and small-angle X-ray scattering. It was confirmed that the as-cast film shows lamellar structure, however, the structure started to transform into Gyroid structure after annealing 14 h and it took more than 2 days to accomplish the transition. Furthermore, it was also confirmed that Gyroid structure is oriented with its (110) normal to the film surface, the length of its repeating unit along with (110), 55 nm, is similar to its original lamellar domain spacing, 58 nm.     

Formation of giant protein vesicles by a lipid cosolvent method

This paper describes a method to create giant protein vesicles (GPVs) of ≥10 μm by solvent-driven fusion of large vesicles (0.1-0.2 μm) with reconstituted membrane proteins. We found that formation of GPVs proceeded from rotational mixing of protein-reconstituted large unilamellar vesicles (LUVs) with a lipid-containing solvent phase. We made GPVs by using n-decane and squalene as solvents, and applied generalized polarization (GP) imaging to monitor the polarity around the protein transmembrane region of aquaporins labeled with the polarity-sensitive probe Badan. Specifically, we created GPVs of spinach SoPIP2;1 and E. coli AqpZ aquaporins. Our findings show that hydrophobic interactions within the bilayer of formed GPVs are influenced not only by the solvent partitioning propensity, but also by lipid composition and membrane protein isoform.     

Bacterial lipid modification of proteins for novel protein engineering applications

Functioning of proteins efficiently at the solid-liquid interface is critical to not only biological but also modern man-made systems such as ELISA, liposomes and biosensors. Anchoring hydrophilic proteins poses a major challenge in this regard. Lipid modification, N-acyl-S-diacylglyceryl-Cys, providing an N-terminal hydrophobic membrane anchor is a viable solution that bacteria have successfully evolved but remains unexploited. Based on the current understanding of this ubiquitous and unique bacterial lipid modification it is possible to use Escherichia coli, the popular recombinant protein expression host, for converting a non-lipoprotein to a lipoprotein with a hydrophobic anchor at the N-terminal end. We report two strategies applicable to non-lipoproteins (with or without signal sequences) employing minimal sequence change. Taking periplasmic Shigella apyrase as an example, its signal sequence was engineered to include a lipobox, an essential determinant for lipid modification, or its mature sequence was fused to the signal sequence of abundant outer membrane lipoprotein, Lpp. Lipid modification was proved by membrane localization, electrophoretic mobility shift and mass spectrometric analysis. Substrate specificity and specific activity measurements indicated functional integrity after modification. In conclusion, a convenient protein engineering strategy for converting non-lipoprotein to lipoprotein for commercial application has been devised and tested successfully.     

Effect of protein source and resistance training on body composition and sex hormones

Background  

Evidence suggests an inverse relationship between soy protein intake and serum concentrations of male sex hormones. Anecdotal evidence indicates that these alterations in serum sex hormones may attenuate changes in lean body mass following resistance training. However, little empirical data exists regarding the effects of soy and milk-based proteins on circulating androgens and exercise induced body composition changes.     

The spherulitic and lamellar morphology of melt-crystallized isotactic polypropylene

A study is presented concerning the basic morphology of melt-crystallized isotactic polypropylene (iPP). Involved within, is the coordinated application of optical and electronmicroscopy on a range of commercial iPP-s, crystallized in the temperature range 100°C–150°C. For electron microscopy in particular, the permanganic etching technique has been used throughout, providing the simultaneous combination of both real space microstructures with electron diffraction information. The investigation itself has centred on the five different spherulite types, as identified optically, which were then correlated with the details of their particular lamellar morphology. It was found that each spherulite type is characterized by virtue of the arrangement of its constituent lamellae, in terms of orientation, habit type and crystal structure. Thus, specific correlations were obtained between the structural entities on all scales of the structure hierarchy.     

Obtaining enzymatic protein and lipid hydrolysates from byproducts of the meat processing industry

Development of methods for recycling the byproducts of food industry is one of the most important areas of biotechnology. Fat- and keratin-containing byproducts (horns, hooves, and feathers of slaughtered animals) are the most valuable because they are rich in biologically active substances; in addition, their processing makes meat processing industry more environmentally pure. The purpose of this study was to develop the methods for increasing the efficiency of enzymatic hydrolysis of industrial substances with technical preparations of hydrolases. The results of the study have been used for developing kinetic models of the processes and determining their optimal conditions ensuring a degree of substrate conversion of at least 95%. Methods for intensification of these enzymatic processes with the use of alkyl hydroxybenzenes have been suggested that allow both the expenditure of the enzymatic preparation (EP) and the duration of enzymatic hydrolysis to be decreased (by a factor of four). Hydrolysis at increased temperatures (55–65°C) decreases the risk of extraneous microflora growth, making it unnecessary to maintain aseptic conditions during hydrolysis and permitting the use of less elaborate equipment. It has been demonstrated that alkyl hydroxy-benzenes can be used to increase the efficiency of enzymatic processes using technical oxidases and hydrolases. The results of the study may be used at meat processing plants to organize the production of highly competitive, valuable products based on enzymatic hydrolysates through recycling the byproducts of the major processes.