Chemical Oxidation (Using Ozone) of the Spore Wall of Lycopodium Clavatum

Spore walls of Lycopodium clavatum, after oxidation using ozone for varying times and treatment with dilute alkali, showed a striking loss of the outer layer (muri) of the exine. Almost all trace of the muri and undetermined portions of the remaining exine wall were removed after 20 hours' oxidation. The exine wall material seems to be evenly removed. The only degradation patterns observed are rare pits after 20 hours oxidation. A degradation-related colour change of the spore wall occurs after 11 to 16 hours' oxidation.     

Molecular Dynamics of Thermoenzymes at High Temperature and Pressure: A Review

Lipases are known for their versatility in addition to their ability to digest fat. They can be used for the formulation of detergents, as food ingredients and as biocatalysts in many industrial processes. Because conventional enzymes are frangible at high temperatures, the replacement of conventional chemical routes with biochemical processes that utilize thermostable lipases is vital in the industrial setting. Recent theoretical studies on enzymes have provided numerous fundamental insights into the structures, folding mechanisms and stabilities of these proteins. The studies corroborate the experimental results and provide additional information regarding the structures that were determined experimentally. In this paper, we review the computational studies that have described how temperature affects the structure and dynamics of thermoenzymes, including the thermoalkalophilic L1 lipase derived from Bacillus stearothermophilus. We will also discuss the potential of using pressure for the analysis of the stability of thermoenzymes because high pressure is also important for the processing and preservation of foods.     

超高静压协同中温对凝结芽孢杆菌芽孢灭活动力学规律的研究

研究了超高静压协同中温对凝结芽孢杆菌芽孢在磷酸缓冲液和牛奶(经超高温灭菌)中灭活的动力学规律, 并对超高静压的升压过程及相应的灭活效果进行了研究。结果表明, 升压过程对凝结芽孢杆菌芽孢灭活的影响不能忽略, 且随压力增加这种效果越强, 最高使其下降1.77个数量级; 凝结芽孢杆菌芽孢在牛奶中比在磷酸缓冲液中有更高的抗性; 在3种拟合模型(线性、Weibull和Log-logistic模型)中, 线性模型不适合模拟这些存活曲线, 而Log-logistic模型能更好地模拟这些存活曲线, 其次是Weibull模型。     

超高静压协同中温对凝结芽孢杆菌芽孢灭活动力学规律的研究

研究了超高静压协同中温对凝结芽孢杆菌芽孢在磷酸缓冲液和牛奶(经超高温灭菌)中灭活的动力学规律,并对超高静压的升压过程及相应的灭活效果进行了研究.结果表明,升压过程对凝结芽孢杆菌芽孢灭活的影响不能忽略,且随压力增加这种效果越强,最高使其下降1.77个数量级;凝结芽孢杆菌芽孢在牛奶中比在磷酸缓冲液中有更高的抗性;在3种拟合模型(线性、Weibull和Log-logistic模型)中,线性模型不适合模拟这些存活曲线,而Log-logistic模型能更好地模拟这些存活曲线,其次是Weibull模型.     

Abiotic Formation of Valine Peptides Under Conditions of High Temperature and High Pressure

We investigated the oligomerization of solid valine and the stabilities of valine and valine peptides under conditions of high temperature (150–200 °C) and high pressure (50–150 MPa). Experiments were performed under non-aqueous condition in order to promote dehydration reaction. After prolonged exposure of monomeric valine to elevated temperatures and pressures, the products were analyzed by liquid chromatography mass spectrometry comparing their retention times and masses. We identified linear peptides that ranged in size from dimer to hexamer, as well as a cyclic dimer. Previous studies that attempted abiotic oligomerization of valine in the absence of a catalyst have never reported valine peptides larger than a dimer. Increased reaction temperature increased the dissociative decomposition of valine and valine peptides to products such as glycine, β-alanine, ammonia, and amines by processes such as deamination, decarboxylation, and cracking. The amount of residual valine and peptide yields was greater at higher pressures at a given temperature, pressure, and reaction time. This suggests that dissociative decomposition of valine and valine peptides is reduced by pressure. Our findings are relevant to the investigation of diagenetic processes in prebiotic marine sediments where similar pressures occur under water-poor conditions. These findings also suggest that amino acids, such as valine, could have been polymerized to peptides in deep prebiotic marine sediments within a few hundred million years.     

银耳子实体多糖高温高压提取工艺研究

通过对银耳子实体多糖高温高压提取工艺中,提取温度、料液比、提取时间,提取次数等影响因素的实验分析,确定银耳子实体多糖高温高压提取的最佳条件为:提取温度110℃(对应压力0.04 MPa);料液比为1∶70;提取时间为2 h;提取次数为2次,在此条件下,银耳子实体多糖提取率可达36.5%。     

Interactions of Oxygen at High Pressure and Radiation in Drosophila

Oxygen at high pressure (OHP) and X-irradiation can interact in the fruit fly Drosophila melanogaster to potentiate toxic actions characteristic of one agent alone. 1000 kvp X-irradiation in doses of 30, 60, and 75 kr accelerated the acute immobilization of young male Drosophila by oxygen at 7.8 atm, up to rates twice that observed with such oxygen pressure alone. X-irradiation alone in these dosages did not acutely immobilize the Drosophila. X-irradiation during exposure to 7.8 atm pO₂ was more effective and consistent in producing this potentiation than was X-irradiation that preceded exposure to OHP. Acute OHP toxicity in young female Drosophila was not potentiated by 75 kr of X-irradiation. On the other hand, shortening of the life span of young male Drosophila by the above doses of X-irradiation was augmented significantly by a concurrent 40 min exposure to OHP (which alone did not significantly decrease life span). This shows, for the first time, that oxygen can affect not only the acute effects of radiation, but also the residual irreversible effects indicated by the life span shortening.     

Topography and Non-Topography in Exine Organization

Table I shows that two letters (S, N) and two figures (1, 2) can be used as symbols or “labels” instead of more or less complicated and/or ambiguous terms in labelling the main topographical and/or non-topographical sub-units or parts of angiosperm exine types (at least as far as present knowledge goes). Special attention has been paid to the characteristics of the nexine in Epilobium montanum believed to be “nexine-2” (N2) by Erdtman and others, albeit classified as “nexine-1, footlayer of the ektexine” by Rowley et al. (1970; p. 7).     

Exine and tapetum development in Symphytum officinale (Boraginaceae). Exine substructure and its interpretation

After detailing the exine ontogeny, our purpose was to find out whether the sequence of sporoderm developmental events corresponds to self-assembling micellar mesophases, initiated by genomically determined physicochemical parameters and induced by surfactant glycoproteins at increasing concentrations. Indeed, a scaffolding of the future exine, i.e., the glycocalyx, initiates with scattered clots, which then appear as clusters of spherical and worm-like micelles, derived from surface-active glycoproteins. At the middle tetrad stage, a continuous layer of the glycocalyx emerges, consisting of parallel, tightly packed cylinder-like units, which we interpret as a layer of cylindrical micelles, the so-called middle mesophase. These units bear dark-contrasted particles, arranged in strings or columns. These sites of the glycocalyx units?Cmicelles accumulate initial sporopollenin, hence the term ??sporopollenin acceptor particles?? (SAPs). This process leads to the appearance of procolumellae at the late tetrad stage. The glycocalyx units are rooted into callose and into the microspore cytoplasm. After formation of the tectum and the foot layer, the endexine initiates as a thin layer, and the latter develops into a very thick layer in the post-tetrad period. When callose disintegrates, ??bouquets?? of SAPs become evident on the tectum, which were evidently hidden inside the callose layer; these structures self-assemble into supratectal gemmae. An unusual, ??hybrid?? type of tapetum was observed. What is observed in Symphytum exine development allows us to obtain more evidence for the hypothesis of the participation of micellar self-assembly in sporoderm development and to bring together the concepts of micelles and of SAPs.     

Induction of Colicin la at High Temperature

Induction of colicin la at a high temperature (42 C) was demonstrated in a wild strain of Shigella sonnei. After transfer of the Col factor, a similar effect was observed in those recipient cells acquiring colicinogeny. The induction is suggested to be due to the presence of a thermosensitive colicin repressor. The wild-type strain segregated sensitive cells and showed heterogeneity in colicin production.     

On the Ultrastructure of Pollen Exine in Metasequoia Hu and Cheng

Metasequoia is endemic to China. Present study deals with ultrastructure of pollen exine of M. glyptostroboides Hu et Cheng, and in comparision with other genera of the family. Pollen grains of Metasequoia are spheroidal or subsphoroidal and 27.8(24.3–32.3) μm in diameter. There is a papilla in the distal face. The papilla is wide at the base, 3.5–5.2 μm high, with pointed and circular end and the base crooked toward one side. Exine is about L5 μm thick, layers distinct, Nexine is as thick as sexine. Surface weakly granulate. According to observation by SEM, exine is covered with fine granules and rather coarse tuberculae. The former can be easily separated from the latter. The loose and uneven tuberculae are provided with minute spinules on the surface and generally fall off after acetolysis. The fine and dense granulae, however, remain intact after acetolysis. The study by TEM shows that ektexine is made of granules densely arranged and connected with each other. In addition, sparse Ubisch bodies are unevenly distributed on granular layer with geminate surface. The thick endexine, is composed of 10–15 lamellae. It is worthy to note that all lamellae possess tripartite structure. But lamellae of endexine in other genera of Taxodiaceae have no tripartite structure except the lamella near ektexine. Number of lamella and thickness of endexine in Metasequoia differ from those of other genera in Taxodiaceae; for example endexine with 8–10 lamellae in Taxodium, 8–9 lamellae in Sequoia, 6–7 lamellae in Glyptostrobus, 6–8 lamellae in Cunninghamia, about 16 lamellae in Cryptomeria etc.     

Vascular architecture in shoots of early divergent vascular plants, Lycopodium clavatum and Lycopodium annotinum

Lycopodium represents a phylogenetically distinct clade of basal vascular plants with anatomical characters that have no parallel in other lineages. Thus, knowledge of lycopod structure and development may reveal important information about the common ancestors of all vascular plants. Here we report the unique architecture of the conducting system in Lycopodium annotinum and Lycopodium clavatum. Based on multiple series of anatomical sections, we reconstructed spatial relationships between microphylls and the stelar system. Analysis revealed that protoxylem ribs (PXR) were vertical, regardless of type of phyllotaxis, and their numbers were variable. Microphyll traces (MTr) were randomly distributed between ribs, resulting in the absence of defined sympodia and varied lengths of MTr. Dichotomous branching contributed to additional features, for example occurrence of mesarch protoxylem, affecting stele structure and PXR numbers. Our data showed limited interrelationships between lycopod vasculature and microphyll phyllotaxis. This may suggest that both systems developed independently, then evolved together to form the integrated supply system. Thus vasculature in extant lycophytes may be less functionally efficient than in seed plants, where consistent leaf-trace lengths guarantee predictable energy utilization during ontogeny. Differences may result from the phylogenetically different origin of microphylls, and the level of vascular complexity.     

Fruiting at High Temperature and Its Genetic Control in the Basidiomycete Flammulina velutipes

Of 10 geographic strains of Flammulina velutipes, 4 were found capable of fruiting at 22°C (Fr_H) rather than at the typical 15°C (Fr_L). Crosses made between Fr_H and Fr_L monokaryons were never observed to fruit at 22°C. However, some hybrids did fruit at the intermediate temperature of 18°C when grown on appropriate substrates, indicating incomplete dominance of the low-temperature requirement. Analysis of progeny of five Fr_H × Fr_L crosses indicated that a minimum of two genes appears to control the requirement for fruiting at ≤15°C. The genes are not closely linked to either incompatibility locus.     

Combined Effects of High Hydrostatic Pressure and Temperature for Inactivation of Bacillus anthracis Spores

Spores of Bacillus anthracis are known to be extremely resistant to heat treatment, irradiation, desiccation, and disinfectants. To determine inactivation kinetics of spores by high pressure, B. anthracis spores of a Sterne strain-derived mutant deficient in the production of the toxin components (strain RP42) were exposed to pressures ranging from 280 to 500 MPa for 10 min to 6 h, combined with temperatures ranging from 20 to 75°C. The combination of heat and pressure resulted in complete destruction of B. anthracis spores, with a D value (exposure time for 90% inactivation of the spore population) of approximately 4 min after pressurization at 500 MPa and 75°C, compared to 160 min at 500 MPa and 20°C and 348 min at atmospheric pressure (0.1 MPa) and 75°C. The use of high pressure for spore inactivation represents a considerable improvement over other available methods of spore inactivation and could be of interest for antigenic spore preparation.     

Imaging Leukocyte Adhesion to the Vascular Endothelium at High Intraluminal Pressure

Worldwide, hypertension is reported to be in approximately a quarter of the population and is the leading biomedical risk factor for mortality worldwide. In the vasculature hypertension is associated with endothelial dysfunction and increased inflammation leading to atherosclerosis and various disease states such as chronic kidney disease², stroke³ and heart failure⁴. An initial step in vascular inflammation leading to atherogenesis is the adhesion cascade which involves the rolling, tethering, adherence and subsequent transmigration of leukocytes through the endothelium. Recruitment and accumulation of leukocytes to the endothelium is mediated by an upregulation of adhesion molecules such as vascular cell adhesion molecule-1 (VCAM-1), intracellular cell adhesion molecule-1 (ICAM-1) and E-selectin as well as increases in cytokine and chemokine release and an upregulation of reactive oxygen species⁵. In vitro methods such as static adhesion assays help to determine mechanisms involved in cell-to-cell adhesion as well as the analysis of cell adhesion molecules. Methods employed in previous in vitro studies have demonstrated that acute increases in pressure on the endothelium can lead to monocyte adhesion, an upregulation of adhesion molecules and inflammatory markers⁶ however, similar to many in vitro assays, these findings have not been performed in real time under physiological flow conditions, nor with whole blood. Therefore, in vivo assays are increasingly utilised in animal models to demonstrate vascular inflammation and plaque development. Intravital microscopy is now widely used to assess leukocyte adhesion, rolling, migration and transmigration^7-9. When combining the effects of pressure on leukocyte to endothelial adhesion the in vivo studies are less extensive. One such study examines the real time effects of flow and shear on arterial growth and remodelling but inflammatory markers were only assessed via immunohistochemistry¹⁰. Here we present a model for recording leukocyte adhesion in real time in intact pressurised blood vessels using whole blood perfusion. The methodology is a modification of an ex vivo vessel chamber perfusion model⁹ which enables real-time analysis of leukocyte -endothelial adhesive interactions in intact vessels. Our modification enables the manipulation of the intraluminal pressure up to 200 mmHg allowing for study not only under physiological flow conditions but also pressure conditions. While pressure myography systems have been previously demonstrated to observe vessel wall and lumen diameter¹¹ as well as vessel contraction this is the first time demonstrating leukocyte-endothelial interactions in real time. Here we demonstrate the technique using carotid arteries harvested from rats and cannulated to a custom-made flow chamber coupled to a fluorescent microscope. The vessel chamber is equipped with a large bottom coverglass allowing a large diameter objective lens with short working distance to image the vessel. Furthermore, selected agonist and/or antagonists can be utilized to further investigate the mechanisms controlling cell adhesion. Advantages of this method over intravital microscopy include no involvement of invasive surgery and therefore a higher throughput can be obtained. This method also enables the use of localised inhibitor treatment to the desired vessel whereas intravital only enables systemic inhibitor treatment.     

Damage in Escherichia coli Cells Treated with a Combination of High Hydrostatic Pressure and Subzero Temperature

The relationship between membrane permeability, changes in ultrastructure, and inactivation in Escherichia coli strain K-12TG1 cells subjected to high hydrostatic pressure treatment at room and subzero temperatures was studied. Propidium iodide staining performed before and after pressure treatment made it possible to distinguish between reversible and irreversible pressure-mediated cell membrane permeabilization. Changes in cell ultrastructure were studied using transmission electron microscopy (TEM), which showed noticeable condensation of nucleoids and aggregation of cytosolic proteins in cells fixed after decompression. A novel technique used to mix fixation reagents with the cell suspension in situ under high hydrostatic pressure (HHP) and subzero-temperature conditions made it possible to show the partial reversibility of pressure-induced nucleoid condensation. However, based on visual examination of TEM micrographs, protein aggregation did not seem to be reversible. Reversible cell membrane permeabilization was noticeable, particularly for HHP treatments at subzero temperature. A correlation between membrane permeabilization and cell inactivation was established, suggesting different mechanisms at room and subzero temperatures. We propose that the inactivation of E. coli cells under combined HHP and subzero temperature occurs mainly during their transiently permeabilized state, whereas HHP inactivation at room temperature is related to a balance of transient and permanent permeabilization. The correlation between TEM results and cell inactivation was not absolute. Further work is required to elucidate the effects of pressure-induced damage on nucleoids and proteins during cell inactivation.     

Monounsaturated but Not Polyunsaturated Fatty Acids Are Required for Growth of the Deep-Sea Bacterium Photobacterium profundum SS9 at High Pressure and Low Temperature

There is considerable evidence correlating the production of increased proportions of membrane unsaturated fatty acids (UFAs) with bacterial growth at low temperatures or high pressures. In order to assess the importance of UFAs to microbial growth under these conditions, the effects of conditions altering UFA levels in the psychrotolerant piezophilic deep-sea bacterium Photobacterium profundum SS9 were investigated. The fatty acids produced by P. profundum SS9 grown at various temperatures and pressures were characterized, and differences in fatty acid composition as a function of phase growth, and between inner and outer membranes, were noted. P. profundum SS9 was found to exhibit enhanced proportions of both monounsaturated (MUFAs) and polyunsaturated (PUFAs) fatty acids when grown at a decreased temperature or elevated pressure. Treatment of cells with cerulenin inhibited MUFA but not PUFA synthesis and led to a decreased growth rate and yield at low temperature and high pressure. In addition, oleic acid-auxotrophic mutants were isolated. One of these mutants, strain EA3, was deficient in the production of MUFAs and was both low-temperature sensitive and high-pressure sensitive in the absence of exogenous 18:1 fatty acid. Another mutant, strain EA2, produced little MUFA but elevated levels of the PUFA species eicosapentaenoic acid (EPA; 20:5n-3). This mutant grew slowly but was not low-temperature sensitive or high-pressure sensitive. Finally, reverse genetics was employed to construct a mutant unable to produce EPA. This mutant, strain EA10, was also not low-temperature sensitive or high-pressure sensitive. The significance of these results to the understanding of the role of UFAs in growth under low-temperature or high-pressure conditions is discussed.     

Tuberization in Potato at High Temperatures: Interaction between Temperature and Irradiance

Potato plants (Solanum tuberosum L., cv. Sebago) responded similarlyto high temperatures and low irradiance by diverting dry matterto the shoots rather than the tubers, and changes were notedin a range of morphological characteristics. It is proposedthat the effect of both high temperature and low irradianceis brought about by the increased production of a growth substance,possibly gibberellin, which inhibits tuber formation, and thattuber yield is determined by the balance between temperatureand irradiance. Solanum tuberosum L., potato, tuberization, temperature, irradiance, gibberellin