Determination of Protein

Elastic and inelastic dark-field micrographs simultaneously recorded by the scanning transmission electron microscope (STEM) provide structural and quantitative chemical information at a spatial resolution of some nanometers. Simultaneous acquisition is essential as it warrants: (i) the geometrical identity of picture elements (pixels) in these micrographs, and (ii) the identity of recording conditions (focus, electron dose) for the elastic and inelastic images. Suitable off-line processing of such multichannel images allows the concentration of protein or nucleic acid within embedding material to be evaluated. In addition, location and number of chemical elements such as phosphorus can be determined. Theoretical sensitivity is a few phosphorus atoms in a volume of approximately 1000 nm3 (not demonstrated in this paper). Quantitative electron microscopy of this kind, however, requires the influence of mass loss as well as plural scattering to be correctly accounted for. Experimental conditions that provide a sound basis for concentration determination as well as element mapping are discussed

Cryo-Preparation and Planar Magnetron Sputtering for Low Temperature Scanning Electron Microscopy

Cryo-preparation is a reliable technique for the structural investigation of food products in low temperature scanning electron microscopy (SEM). Artifacts, such as, the segregation of water/non-water ingredients, occur during the freezing process by the crystallization of ice; they can be helpful for correct interpretation of visualized details, e.g., the detection of water containing compartments. The size of the segregation structures depends on water concentration and specimen thickness. The condensation of water vapor (ice contamination) is influenced by the specimen temperature and the partial pressure of the water inside the vacuum system. Furthermore, the evaporation (sublimation, etching) of specimen water can be regulated by monitoring the specimen temperature. Sublimation under SEM observation, i.e., in situ etching at low acceleration voltage, allows the progress of etching to be observed continuously, prior to the coating of the specimen inside a dedicated cryo-preparation system attached to the SEM. Coating of specimens provides superior structural resolution compared with the observation of uncoated samples. A coating layer of platinum ( ~ 1-2 nm thick), deposited on a cold substrate by planar magnetron sputtering, is almost homogenous and has a density close to that of the solid metal. Its use allows bulk biological specimens to be observed in low temperature SEM with a structural resolution up to the visualization of transmembrane proteins

Rhizobia contribute to salinity tolerance in common beans (Phaseolus vulgaris L.)

Rhizobia are soil bacteria that induce nodule formation on leguminous plants. In the nodules, they reduce dinitrogen to ammonium that can be utilized by plants. Besides nitrogen fixation, rhizobia have other symbiotic functions in plants including phosphorus and iron mobilization and protection of the plants against various abiotic stresses including salinity. Worldwide, about 20% of cultivable and 33% of irrigation land is saline, and it is estimated that around 50% of the arable land will be saline by 2050. Salinity inhibits plant growth and development, results in senescence, and ultimately plant death. The purpose of this study was to investigate how rhizobia, isolated from Kenyan soils, relieve common beans from salinity stress. The yield loss of common bean plants, which were either not inoculated or inoculated with the commercial R. tropici rhizobia CIAT899 was reduced by 73% when the plants were exposed to 300 mM NaCl, while only 60% yield loss was observed after inoculation with a novel indigenous isolate from Kenyan soil, named S3. Expression profiles showed that genes involved in the transport of mineral ions (such as K+, Ca2+, Fe3+, PO43−, and NO3−) to the host plant, and for the synthesis and transport of osmotolerance molecules (soluble carbohydrates, amino acids, and nucleotides) are highly expressed in S3 bacteroids during salt stress than in the controls. Furthermore, genes for the synthesis and transport of glutathione and γ-aminobutyric acid were upregulated in salt-stressed and S3-inocculated common bean plants. We conclude that microbial osmolytes, mineral ions, and antioxidant molecules from rhizobia enhance salt tolerance in common beans

A report on the study of land-sourced pollutants and their impacts on water quality in and adjacent to the Great Barrier Reef: an assessment to guide the development of management plans to halt any decline in the water quality of river catchments draining to the Reef, as a result of land-based pollution, and to achieve the long-term goal of reversing any trend in declining water quality

Includes 'Summary statement of the Reef Science Panel regarding water quality in and adjacent to the Great Barrier Reef