Phenotypic characterization of Synechocystis sp. PCC 6803 substrains reveals differences in sensitivity to abiotic stress

© 2017 Zavřel et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Synechocystis sp. PCC 6803 is a widely used model cyanobacterium, whose substrains can vary on both genotype and phenotype levels. Previously described phenotypic variations include ability of mixotrophic growth, ability of movement on agar plates and variations in pigments composition or cell size. In this study, we report for the first time significant variation among Synechocystis substrains in complex cellular traits such as growth rate, photosynthesis efficiency, cellular dry weight and cellular composition (including protein or carbohydrates content). We also confirmed previously reported differences in cell size. Synechocystis cultures were cultivated in controlled environment of flat panel photobioreactors under red, blue and white light of intensities up to 790 μmol(photons) m-2 s-1, temperatures 23C–60C, input CO2 concentrations ranging from 400 to 15 000 ppm and in BG11 cultivation medium with and without addition of NaCl. Three Synechocystis substrains were used for the comparative experiments: GT-L, GT-B (Brno, CZ) and PCC-B (Brno, CZ). Growth rates of Synechocystis GT-B were inhibited under high intensities of red light (585–670 nm), and growth rates of both substrains GT-B and PCC-B were inhibited under photons of wavelengths 485–585 nm and 670–700 nm. Synechocystis GT-B was more sensitive to low temperatures than the other two tested substrains, and Synechocystis GT-L was sensitive to the presence of NaCl in the cultivation media. The results suggest that stress sensitivity of commonly used Synechocystis substrains can strongly vary, similarly as glucose tolerance or motility as reported previously. Our study further supports the previous statement that emphasizes importance of proper Synechocystis substrains selection and awareness of phenotypical differences among Synechocystis substrains which is crucial for comparative and reproducible research. This is highly relevant for studies related to stress physiology and development of sustainable biotechnological applications

Optimizing cyanobacterial product synthesis: Meeting the challenges.

The synthesis of renewable bioproducts using photosynthetic microorganisms holds great promise. Sustainable industrial applications, however, are still scarce and the true limits of phototrophic production remain unknown. One of the limitations of further progress is our insufficient understanding of the quantitative changes in photoautotrophic metabolism that occur during growth in dynamic environments. We argue that a proper evaluation of the intra- and extracellular factors that limit phototrophic production requires the use of highly-controlled cultivation in photobioreactors, coupled to real-time analysis of production parameters and their evaluation by predictive computational models. In this addendum, we discuss the importance and challenges of systems biology approaches for the optimization of renewable biofuels production. As a case study, we present the utilization of a state-of-the-art experimental setup together with a stoichiometric computational model of cyanobacterial metabolism for quantitative evaluation of ethylene production by a recombinant cyanobacterium Synechocystis sp. PCC 6803

Can algal biotechnology bring effective solution for closing the phosphorus cycle? Use of algae for nutrient removal - Review of past trends and future perspectives in the context of nutrient recovery

Eutrophication of water by nutrient pollution is a global environmental issue. Biological methods for removing nutrients are environmentally friendly and sustainable. Therefore, this article summarizes main trends in the use of algae for removing nutrients from wastewater using both suspended and attached algal-based systems. A wide variety of algal species and experimental approaches has been tested to date. Researchers report that algae are able to effectively remove a variety of pollutants and nutrients. This review also discusses the potential of algal-based technology for nutrient, especially phosphorus, recovery. Despite the fact that effective nutrient removal has been demonstrated, there are still many challenges to be overcome in the development of succesfull technologies

Further support for the alignment of cattle along magnetic field lines: reply to Hert et al.

Hert et al. (J Comp Physiol A, 2011) challenged one part of the study by Begall et al. (PNAS 105:13451–13455, 2008) claiming that they could not replicate the finding of preferential magnetic alignment of cattle recorded in aerial images of Google Earth. However, Hert and co-authors used a different statistical approach and applied the statistics on a sample partly unsuitable to examine magnetic alignment. About 50% of their data represent noise (resolution of the images is too poor to enable unambiguous measurement of the direction of body axes, pastures are on slopes, near settlements or high voltage power-lines, etc.). Moreover, the authors have selected for their analysis only ~ 40% of cattle that were present on the pastures analyzed. Here, we reanalyze all usable data and show that cattle significantly align their body axes in North–South direction on pastures analyzed by Hert and co-authors. This finding thus supports our previous study. In addition, we show by using aerial Google Earth images with good resolution, that the magnetic alignment is more pronounced in resting than in standing cattle

Fermat Principle in Finsler Spacetimes

It is shown that, on a manifold with a Finsler metric of Lorentzian signature, the lightlike geodesics satisfy the following variational principle. Among all lightlike curves from a point (emission event) to a timelike curve (worldline of receiver), the lightlike geodesics make the arrival time stationary. Here ``arrival time'' refers to a parametrization of the timelike curve. This variational principle can be applied (i) to the vacuum light rays in an alternative spacetime theory, based on Finsler geometry, and (ii) to light rays in an anisotropic non-dispersive medium with a general-relativistic spacetime as background.Comment: 18 pages, submitted to Gen. Rel. Gra

Quantitative insights into the cyanobacterial cell economy

© Zavřel et al. Phototrophic microorganisms are promising resources for green biotechnology. Compared to heterotrophic microorganisms, however, the cellular economy of phototrophic growth is still insufficiently understood. We provide a quantitative analysis of light-limited, light-saturated, and light-inhibited growth of the cyanobacterium Synechocystis sp. PCC 6803 using a reproducible cultivation setup. We report key physiological parameters, including growth rate, cell size, and photosynthetic activity over a wide range of light intensities. Intracellular proteins were quantified to monitor proteome allocation as a function of growth rate. Among other physiological acclimations, we identify an upregulation of the translational machinery and downregulation of light harvesting components with increasing light intensity and growth rate. The resulting growth laws are discussed in the context of a coarse-grained model of phototrophic growth and available data obtained by a comprehensive literature search. Our insights into quantitative aspects of cyanobacterial acclimations to different growth rates have implications to understand and optimize photosynthetic productivity

Effects of a red card on goal-scoring in World Cup football matches

We examine the effect of the sending-off of a player on the goal-scoring rates in FIFA World Cup matches in tournaments from 1998 to 2014. We use a hazard rate framework in which the effect of a red card is modeled as a shift in the goal-scoring rate. A red card may harm the team that receives a red card and may be beneficial for their opponent. Indeed, we find that the goal-scoring rate of the sanctioned team goes down, while the goal-scoring rate of the non-sanctioned team goes up

The CHK1 inhibitor MU380 significantly increases the sensitivity of human docetaxel-resistant prostate cancer cells to gemcitabine through the induction of mitotic catastrophe.

As treatment options for patients with incurable metastatic castration-resistant prostate cancer (mCRPC) are considerably limited, novel effective therapeutic options are needed. Checkpoint kinase 1 (CHK1) is a highly conserved protein kinase implicated in the DNA damage response (DDR) pathway that prevents the accumulation of DNA damage and controls regular genome duplication. CHK1 has been associated with prostate cancer (PCa) induction, progression, and lethality; hence, CHK1 inhibitors SCH900776 (also known as MK-8776) and the more effective SCH900776 analog MU380 may have clinical applications in the therapy of PCa. Synergistic induction of DNA damage with CHK1 inhibition represents a promising therapeutic approach that has been tested in many types of malignancies, but not in chemoresistant mCRPC. Here, we report that such therapeutic approach may be exploited using the synergistic action of the antimetabolite gemcitabine (GEM) and CHK1 inhibitors SCH900776 and MU380 in docetaxel-resistant (DR) mCRPC. Given the results, both CHK1 inhibitors significantly potentiated the sensitivity to GEM in a panel of chemo-naïve and matched DR PCa cell lines under 2D conditions. MU380 exhibited a stronger synergistic effect with GEM than clinical candidate SCH900776. MU380 alone or in combination with GEM significantly reduced spheroid size and increased apoptosis in all patient-derived xenograft 3D cultures, with a higher impact in DR models. Combined treatment induced premature mitosis from G1 phase resulting in the mitotic catastrophe as a prestage of apoptosis. Finally, treatment by MU380 alone, or in combination with GEM, significantly inhibited tumor growth of both PC339-DOC and PC346C-DOC xenograft models in mice. Taken together, our data suggest that metabolically robust and selective CHK1 inhibitor MU380 can bypass docetaxel resistance and improve the effectiveness of GEM in DR mCRPC models. This approach might allow for dose reduction of GEM and thereby minimize undesired toxicity and may represent a therapeutic option for patients with incurable DR mCRPC

Quantifying Cyanothece growth under DIC limitation.

The photoautotrophic, unicellular N2-fixer, Cyanothece, is a model organism that has been widely used to study photosynthesis regulation, the structure of photosystems, and the temporal segregation of carbon (C) and nitrogen (N) fixation in light and dark phases of the diel cycle. Here, we present a simple quantitative model and experimental data that together, suggest external dissolved inorganic carbon (DIC) concentration as a major limiting factor for Cyanothece growth, due to its high C-storage requirement. Using experimental data from a parallel laboratory study as a basis, we show that after the onset of the light period, DIC was rapidly consumed by photosynthesis, leading to a sharp drop in the rate of photosynthesis and C accumulation. In N2-fixing cultures, high rates of photosynthesis in the morning enabled rapid conversion of DIC to intracellular C storage, hastening DIC consumption to levels that limited further uptake. The N2-fixing condition allows only a small fraction of fixed C for cellular growth since a large fraction was reserved in storage to fuel night-time N2 fixation. Our model provides a framework for resolving DIC limitation in aquatic ecosystem simulations, where DIC as a growth-limiting factor has rarely been considered, and importantly emphasizes the effect of intracellular C allocation on growth rate that varies depending on the growth environment

Optimum and standard beam widths for numerical modeling of interface scattering problems

Author Posting. © Acoustical Society of America, 2000. This article is posted here by permission of Acoustical Society of America for personal use, not for redistribution. The definitive version was published in Journal of the Acoustical Society of America 107 (2000): 1095-1102, doi:10.1121/1.428399.Gaussian beams provide a useful insonifying field for surface or interface scattering problems such as encountered in electromagnetics, acoustics and seismology. Gaussian beams have these advantages: (i) They give a finite size for the scattering region on the interface. (ii) The incident energy is restricted to a small range of grazing angles. (iii) They do not have side lobes. (iv) They have a convenient mathematical expression. The major disadvantages are: (i) Insonification of an interface is nonuniform. The scattered field will depend on the location of the scatterers within the beam. (ii) The beams spread, so that propagation becomes an integral component of the scattering problem. A standard beam parameterization is proposed which keeps propagation effects uniform among various models so that the effects of scattering only can be compared. In continuous wave problems, for a given angle of incidence and incident amplitude threshold, there will be an optimum Gaussian beam which keeps the insonified area as small as possible. For numerical solutions of pulse beams, these standard parameters provide an estimate of the smallest truncated domain necessary for a physically meaningful result.This work was carried out under Office of Naval Research Grant Nos. N00014-90-I-1493, N00014-96-1-0460, and N00014-95-1-0506 and under a Mellon Independent Study Award from Woods Hole Oceanographic Institution