Sensitive Detection of the Leaching-out from Membrane-surrounded Compartments: a New Approach Explored with Cells and Tissues of Nicotiana tabacum L.

In search of a simple method for testing the very early events of ozone damage to susceptible plants as well as complete destruction after threshold-exceeding treatments an over-all measurement of cell ingredients by their optical density in the UVB and UVC range was investigated. The cell particles were liberated after membrane permeabilization or after cell bursting. Uncontrolled results could be excluded. Furthermore, the results of the developed spectrophotometrical test could be, in the case of tissue samples (leaf discs), very well reproduced with an osmometrical measurement. The latter was less sensitive and not suitable for cellular samples because the protoplasts must be dissolved in a nearly isotonic medium which caused too large a background for this method but not for the UVS test. Contrary to the osmometric measurement, the photometric one cannot be used for determining the absolute amount of cell ingredients but only for relative measurement between samples in a given range of concentrations. Oxidative changes of the liberated ingredients do not influence their detection, which was demonstrated with ascorbate. The developed leaching test was also useful for determining the membrane damage caused by the detergent Triton X-100, although this was known to have UV absorbance by itself. It was noted that the far UV maximum is not only caused by absorbance and scattering is discussed as an additional explanation.     

Seasonal differences in mineral content, distribution and leakage of sweet pepper seeds

Pepper seed quality is sensitive to variations in climatic conditions during seed development, which might be associated with accumulation, distribution and leakage of mineral elements from the seeds. This was examined in hybrid seeds of sweet pepper (Capsicum annuum L. cv.‘Hazera’ 1195) in two experiments during two growing seasons. The mean daily temperature (day/night) and daily total radiation receipt during seed development were 27.9/23.2°C and 8.63 kW m^?2 in the summer and 18.3/ 14.9°C and 3.18 kW m^?2 in the winter, respectively. Seeds developed in the summer had lower percentage of seedling emergence and leaked a larger portion (45%) of their K content into the water medium than in winter‐developed seeds. Summer seeds accumulated more K and Cl, but less P, Mg, Ca and the weight ratio of linoleic acid to oleic acid was lower than in the winter seeds. The season did not significantly affect N, S and total fatty acids. The most abundant element on the seed coat surface was K in the summer and Ca in the winter seeds. The cotyledon and endosperm of the summer seeds contained relatively higher ratios of K and Ca and lower ratios of P and Mg than the winter seeds. Transportation of mineral nutrients appeared to be involved in the effect of heat and moisture stresses on emergence quality of the pepper seeds.     

Desiccation tolerance in developing soybean seeds: The role of stress proteins

The consistent correlation between desiccation tolerance in orthodox seed tissue and an accumulation of certain "late embryogenesis abundant" (LEA) proteins suggests that these proteins reduce desiccation-induced cellular damage. The aim of the present work was to test this hypothesis. Exogenous abscisic acid (ABA) was used to elevate the level of heal-soluble LEA-like proteins in axes from immature (30 days after flowering: mid-development) seeds of soybean ( Glycine max [L.] Merrill cv. Chippewa 64). As the LEA-like proteins accumulated in response to ABA, the leakage of all elements after desiccation and subsequent rehydration markedly declined. Both LEA-like protein accumulation and the decline in desiccation-induced electrolyte leakage were apparently dependent on the presence of ABA. Both effects of ABA were inhibited by cycloheximide. Light microscopy revealed a marked effect of the ABA on cellular integrity following desiccation. Osmotic stress also caused a decrease in desiccation-induced electrolyte leakage and stimulated the accumulation of LEA-like proteins. Our data are consistent with the hypothesis that the LEA-like proteins contribute to the increase in desiccation tolerance in response to ABA, and are consistent with a general protective role for these proteins in desiccation tolerance.     

Evaluation of quenching methods for metabolite recovery in photoautotrophic Synechococcus sp. PCC 7002

The first step of many metabolomics studies is quenching, a technique vital for rapidly halting metabolism and ensuring that the metabolite profile remains unchanging during sample processing. The most widely used approach is to plunge the sample into prechilled cold methanol; however, this led to significant metabolite loss in Synecheococcus sp. PCC 7002. Here we describe our analysis of the impacts of cold methanol quenching on the model marine cyanobacterium Synechococcus sp. PCC 7002, as well as our brief investigation of alternative quenching methods. We tested several methods including cold methanol, cold saline, and two filtration approaches. Targeted central metabolites were extracted and metabolomic profiles were generated using liquid chromatography-tandem mass spectrometry (LC-MS/MS). The results indicate that cold methanol quenching induces dramatic metabolite leakage in Synechococcus, resulting in a majority of central metabolites being lost prior to extraction. Alternatively, usage of a chilled saline quenching solution mitigates metabolite leakage and improves sample recovery without sacrificing rapid quenching of cellular metabolism. Finally, we illustrate that metabolite leakage can be assessed, and subsequently accounted for, in order to determine absolute metabolite pool sizes; however, our results show that metabolite leakage is inconsistent across various metabolite pools and therefore must be determined for each individually measured metabolite.     

Aerosol generation using nanometer liposome suspensions for pulmonary drug delivery applications

Abstract

Pulmonary lung targeting finds applications in drug delivery to the lung itself and to other body organs, via blood circulation following transfer across alveolar membranes. Understanding pulmonary drug delivery systems towards improving their efficacy needs identification of particle sizes of relevance and elucidation of links between suspension properties, techniques of atomisation and properties of the generated aerosols. This review article is focussed on understanding the elements of pulmonary drug delivery, specifically related to suspensions of small liposomes. Specific objectives of this review include (a) understanding aerosol particle deposition and absorption on pulmonary surface, (b) links between properties of aerosol generation and colloidal drug carriers used for drug encapsulation, and (c) investigation on the controlled properties of liposome aerosols generated using different atomisation techniques for efficacious aerosol therapy.     

Reduction of molecular gas diffusion through gaskets in leaf gas exchange cuvettes by leaf‐mediated pores

There is an ongoing debate on how to correct leaf gas exchange measurements for the unavoidable diffusion leakage that occurs when measurements are done in non‐ambient CO₂ concentrations. In this study, we present a theory on how the CO₂ diffusion gradient over the gasket is affected by leaf‐mediated pores (LMP) and how LMP reduce diffusive exchange across the gaskets. Recent discussions have so far neglected the processes in the quasi‐laminar boundary layer around the gasket. Counter intuitively, LMP reduce the leakage through gaskets, which can be explained by assuming that the boundary layer at the exterior of the cuvette is enriched with air from the inside of the cuvette. The effect can thus be reduced by reducing the boundary layer thickness. The theory clarifies conflicting results from earlier studies. We developed leaf adaptor frames that eliminate LMP during measurements on delicate plant material such as grass leaves with circular cross section, and the effectiveness is shown with respiration measurements on a harp of Deschampsia flexuosa leaves. We conclude that the best solution for measurements with portable photosynthesis systems is to avoid LMP rather than trying to correct for the effects.     

Salicylic acid-induced physiological and biochemical changes in lemongrass varieties under water stress

Abstract

Salicylic acid (SA) treatment reduces the damaging action by water deficit on growth and accelerates a restoration of growth processes. The aim of the present work was to study the physiological and biochemical alteration induced by SA in lemongrass plants under stress conditions. Therefore, a pot culture experiment was conducted to test whether SA application at concentration of (10^?5 M) through foliar spray could protect lemongrass (Cymbopogon flexuosus Steud. Wats.) varieties (Neema and Krishna), subjected to drought stress on the basis of growth parameters and biochemical constituents, proline metabolism and quality attributes including citral content. The treatments were as follows: (i) 100% FC + 0 SA; (ii) 75% FC + 0 SA; (iii) 50% FC + 0 SA; (iv) 75% FC + 10^?5 M SA; and (v) 50% FC + 10^?5 M SA. The growth parameters were significantly reduced under the applied water stress levels; however, foliar application of salicylic acid (10^?5 M) improved the growth parameters in stress-affected plants. The plants under water stress exhibited a significant increase in activities of nitrate reductase and carbonic anhydrase, and electrolyte leakage, proline content, free amino acid and in PEP carboxylase activity. Content and yield of essential oil also significantly decreased in plants that faced water stress. Thus, it was concluded that variety Neema is the more tolerant variety as compared to Krishna on the basis of content and oil yield and well adapted to drought stress conditions.     

On mitigating emissions leakage under biofuel policies

A reason for much pessimism about the environmental benefits of today's biofuels, essentially corn and sugarcane ethanol, is the so‐called indirect land‐use change (ILUC) emissions associated with expanding biofuel production. While there exist several simulation‐based estimates of indirect emissions, the empirical basis underlying key input parameters to such simulations is not beyond doubt, while empirical verification of indirect emissions is hard. Regardless, regulators have adopted global warming intensity ratings for biofuels based on those simulations and in some case are holding regulated firms accountable for (some forms of) leakage. Suffice to say that both the estimates of and the approach to regulating leakage are controversial. The objective of this study is therefore to review a wider economic in order to identify a broader set of policy options for mitigating emissions leakage. We find that controlling leakage by affixing responsibility to regulated firms lacks support in the broader literature, which emphasizes alternative approaches.     

Detecting cell lysis using viscosity monitoring in E. coli fermentation to prevent product loss

Monitoring the physical or chemical properties of cell broths to infer cell status is often challenging due to the complex nature of the broth. Key factors indicative of cell status include cell density, cell viability, product leakage, and DNA release to the fermentation broth. The rapid and accurate prediction of cell status for hosts with intracellular protein products can minimise product loss due to leakage at the onset of cell lysis in fermentation. This article reports the rheological examination of an industrially relevant E. coli fermentation producing antibody fragments (Fab'). Viscosity monitoring showed an increase in viscosity during the exponential phase in relation to the cell density increase, a relatively flat profile in the stationary phase, followed by a rapid increase which correlated well with product loss, DNA release and loss of cell viability. This phenomenon was observed over several fermentations that a 25% increase in broth viscosity (using induction‐point viscosity as a reference) indicated 10% product loss. Our results suggest that viscosity can accurately detect cell lysis and product leakage in postinduction cell cultures, and can identify cell lysis earlier than several other common fermentation monitoring techniques. This work demonstrates the utility of rapidly monitoring the physical properties of fermentation broths, and that viscosity monitoring has the potential to be a tool for process development to determine the optimal harvest time and minimise product loss. © 2016 The Authors. Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers, 32:1069–1076, 2016     

椎体静脉稀疏区注入骨水泥对PVP术中骨水泥渗漏的影响

目的:探讨椎体静脉稀疏区注入骨水泥对骨质疏松椎体压缩性骨折患者行经皮穿刺椎体成形术(percutaneous vertebroplasty,PVP)术中骨水泥渗漏的影响。方法:选择西安交通大学第二附属医院2014年1月至2018年6月收治的61例骨质疏松椎体压缩性骨折患者,根据骨水泥注入区域的不同,将所有患者分为A组(30例)及B组(31例),A组骨水泥注入区域为椎体静脉密集区(椎体中1/3平面处),B组骨水泥注入区域为椎体静脉稀疏区(椎体上1/3及下1/3平面处),对比两组的骨水泥渗漏率,术前、术后6个月时的视觉模拟评分(Visual analogue scale,VAS),治疗中的骨水泥用量、椎体高度恢复率及cobb角恢复度数。结果:B组的骨水泥渗漏率及骨水泥用量均明显低于A组(P0.05)。两组的VAS评分、椎体高度恢复率、cobb角恢复情况对比差异无统计学意义(P0.05)。结论:与椎体静脉密集区相比,在椎体静脉稀疏区注入骨水泥可显著降低骨质疏松椎体压缩性骨折患者PVP术中骨水泥渗漏率,椎体静脉稀疏区可作为PVP术中骨水泥注射的一个相对安全区域。