UV Disinfection sensitivity index of spores or protozoa: A model to predict the required fluence of spores or protozoa.

During UV disinfection, the required UV dose in terms of fluence depends upon the species of bacteria spore and protozoa. To rank their UV disinfection sensitivity, spore sensitivity index (SPSI) and protozoan sensitivity index (PSI) are defined. For spores, shoulder effect exists, therefore, SPSI is defined as the ratio between the ki of any spores for the linear portion of the dose response curve to the kir of Bacillus subtilis as the reference spore. After statistical analysis, the fluence of any spore can be predicted by SPSI through equation, H = (0.8358 ± 0.126)*LogI*SPSI + H0. PSI is defined as the ratio between the inactivation rate constants of a protozoa in reference to that of Cryptosporidium parvum. The equation predicting the fluence of any protozoa in reference to Cryptosporidium parvum is: H = 107.45*(3.86 ± 2.68)*LogI/PSI. Two regression equations suggest that protozoa require significantly higher UV dose than bacteria spores.

The origin of SPA reveals the divergence and convergence of light signaling in Archaeplastida.

Plants have evolved various photoreceptors to adapt to changing light environments, and photoreceptors can inactivate the large CONSTITUTIVE PHOTOMORPHOGENIC/DE-ETIOLATED/FUSCA (COP/DET/FUS) protein complex to release their repression of photoresponsive transcription factors. Here, we tracked the origin and evolution of COP/DET/FUS in Archaeplastida and found that most components of COP/DET/FUS were highly conserved. Intriguingly, the COP1-SUPPRESSOR OF PHYA-105 (SPA) protein originated in Chlorophyta but subsequently underwent a distinct evolutionary history in Viridiplantae. SPA experienced duplication events in the ancestors of specific clades after the colonization of land by plants and was divided into two clades (clades A and B) within euphyllophytes (ferns and seed plants). Our phylogenetic and experimental evidences support a new evolutionary model to clarify the divergence and convergence of light signaling during plant evolution.

Light affects picocyanobacterial grazing and growth response of the mixotrophic flagellate Poterioochromonas malhamensis.

We measured the grazing and growth response of the mixotrophic chrysomonad flagellate Poterioochromonas malhamensis on four closely related picocyanobacterial strains isolated from subalpine lakes in central Europe. The picocyanobacteria represented different pigment types (phycoerythrin-rich, PE, and phycocyanin-rich, PC) and phylogenetic clusters. The grazing experiments were conducted with laboratory cultures acclimated to 10 µmol photon/m2/sec (low light, LL) and 100 µmol photon/m2/sec (moderate light, ML), either in the dark or at four different irradiances ranging from low (6 µmol photon/m2/sec) to high (1,500 µmol photon/m2/sec) light intensity. Poterioochromonas malhamensis preferred the larger, green PC-rich picocyanobacteria to the smaller, red PE-rich picocyanobacterial, and heterotrophic bacteria. The feeding and growth rates of P. malhamensis were sensitive to the actual light conditions during the experiments; the flagellate performed relatively better in the dark and at LL conditions than at high light intensity. In summary, our results found strain-specific ingestion and growth rates of the flagellate; an effect of the preculturing conditions, and, unexpectedly, a direct adverse effect of high light levels. We conclude that this flagellate may avoid exposure to high surface light intensities commonly encountered in temperate lakes during the summer.

DNA-protein cross-links: Formidable challenges to maintaining genome integrity.

DNA is associated with proteins that are involved in its folding and transaction processes. When cells are exposed to chemical cross-linking agents or free radical-generating ionizing radiation, DNA-associated proteins are covalently trapped within the DNA to produce DNA-protein cross-links (DPCs). DPCs produced by these agents contain cross-linked proteins in an undisrupted DNA strand. Some DNA-metabolizing enzymes that form covalent reaction intermediates can also be irreversibly trapped in the presence of inhibitors or DNA damage to give rise to abortive DPCs. The abortive DPCs often contain cross-linked proteins attached to the 5' or 3' end of a DNA strand break. In vitro studies show that steric hindrance caused by cross-linked proteins impedes the progression of DNA helicases and polymerases and of RNA polymerases. The modes and consequences by which DPCs impede replication and transcription processes are considerably different from those with conventional DNA lesions. Thus, DPCs are formidable challenges to maintaining genome integrity and faithful gene expression. Current models of DPC repair involve direct and indirect removal of DPCs. The direct mechanism works for DPCs that contain topoisomerase 2 attached to the 5' end of DNA. The Mre11-Rad50-Nbs1 complex cleaves the site internal to the DPC and directly releases a DPC-containing oligonucleotide. The indirect mechanism involves degradation of cross-linked proteins by proteasomes or the recently identified DPC proteases Wss1 and Sprtn to relieve steric hindrance of DPCs. The resulting peptide-cross-links might be processed by translesion synthesis or other canonical repair mechanisms: however, the exact mechanism remains to be elucidated.

DNA protein crosslink proteolysis repair: From yeast to premature ageing and cancer in humans.

DNA-protein crosslinks (DPCs) are a specific type of DNA lesion consisting of a protein covalently and irreversibly bound to DNA, which arise after exposure to physical and chemical crosslinking agents. DPCs can be bulky and thereby pose a barrier to DNA replication and transcription. The persistence of DPCs during S phase causes DNA replication stress and genome instability. The toxicity of DPCs is exploited in cancer therapy: many common chemotherapeutics kill cancer cells by inducing DPC formation. Recent work from several laboratories discovered a specialized repair pathway for DPCs, namely DPC proteolysis (DPCP) repair. DPCP repair is carried out by replication-coupled DNA-dependent metalloproteases: Wss1 in yeast and SPRTN in metazoans. Mutations in SPRTN cause premature ageing and liver cancer in humans and mice; thus, defective DPC repair has great clinical ramifications. In the present review, we will revise the current knowledge on the mechanisms of DPCP repair and on the regulation of DPC protease activity, while highlighting the most significant unresolved questions in the field. Finally, we will discuss the impact of faulty DPC repair on disease and cancer therapy.

ROS-dependent signalling pathways in plants and algae exposed to high light: Comparisons with other eukaryotes.

Like all aerobic organisms, plants and algae co-opt reactive oxygen species (ROS) as signalling molecules to drive cellular responses to changes in their environment. In this respect, there is considerable commonality between all eukaryotes imposed by the constraints of ROS chemistry, similar metabolism in many subcellular compartments, the requirement for a high degree of signal specificity and the deployment of thiol peroxidases as transducers of oxidising equivalents to regulatory proteins. Nevertheless, plants and algae carry out specialised signalling arising from oxygenic photosynthesis in chloroplasts and photoautotropism, which often induce an imbalance between absorption of light energy and the capacity to use it productively. A key means of responding to this imbalance is through communication of chloroplasts with the nucleus to adjust cellular metabolism. Two ROS, singlet oxygen (1O2) and hydrogen peroxide (H2O2), initiate distinct signalling pathways when photosynthesis is perturbed. 1O2, because of its potent reactivity means that it initiates but does not transduce signalling. In contrast, the lower reactivity of H2O2 means that it can also be a mobile messenger in a spatially-defined signalling pathway. How plants translate a H2O2 message to bring about changes in gene expression is unknown and therefore, we draw on information from other eukaryotes to propose a working hypothesis. The role of these ROS generated in other subcellular compartments of plant cells in response to HL is critically considered alongside other eukaryotes. Finally, the responses of animal cells to oxidative stress upon high irradiance exposure is considered for new comparisons between plant and animal cells.

Production of lutein, and polyunsaturated fatty acids by the acidophilic eukaryotic microalga Coccomyxa onubensis under abiotic stress by salt or ultraviolet light.

In this study, the effect of abiotic stress on the acidophilic eukaryotic microalga, Coccomyxa onubensis, was analyzed for the production of lutein and PUFAs (polyunsaturated fatty acids). It grows autotrophically at a pH of 2.5. It showed a growth rate of 0.30 d-1, and produced approximately 122.50 mg·L-1·d-1 biomass, containing lipids (300.39 mg g-1dw), lutein (5.30 mg g-1dw), and ß-carotene (1.20 mg g-1dw). The fatty acid methyl ester (FAME) fraction was 89.70 mg g-1dw with abundant palmitic acid (28.70%) and linoleic acid (37.80%). The addition of 100 mM NaCl improved the growth rate (0.54 d-1), biomass productivity (243.75 mg·L-1·d-1), and lipids accumulation (416.16 mg g-1dw). The microalga showed a lutein content of 6.70 mg g-1dw and FAME fraction of 118.90 mg g-1dw; 68% of the FAMEs were PUFAs. However, when 200-500 mM salt was added, its growth was inhibited but there was a significant induction of lutein (up to 7.80 mg g-1dw). Under continuous illumination with PAR (photosynthetically active radiations) +UVA (ultraviolet A, 8.7 W m-2), C. onubensis showed a growth rate of 0.40 d-1, and produced 226.3 mg·L-1·d-1 biomass, containing lipids, (487.26 mg g-1dw), lutein (7.07 mg g-1dw), and FAMEs (232.9 mg g-1dw); 48.4% of the FAME were PUFAs. The illumination with PAR + UVB (ultraviolet B, 0.16 W m-2) was toxic for cells. These results indicate that C. onubensis biomass is suitable as a supplement for functional foods and/or source of high added value products.

Ammonium removal using algae-bacteria consortia: the effect of ammonium concentration, algae biomass, and light.

In this study, the effects of ammonium nitrogen concentration, algae biomass concentration, and light conditions (wavelength and intensity) on the ammonium removal efficiency of algae-bacteria consortia from wastewater were investigated. The results indicated that ammonium concentration and light intensity had a significant impact on nitrification. It was found that the highest ammonia concentration (430 mg N/L) in the influent resulted in the highest ammonia removal rate of 108 ± 3.6 mg N/L/days, which was two times higher than the influent with low ammonia concentration (40 mg N/L). At the lowest light intensity of 1000 Lux, algae biomass concentration, light wavelength, and light cycle did not show a significant effect on the performance of algal-bacterial consortium. Furthermore, the ammonia removal rate was approximately 83 ± 1.0 mg N/L/days, which was up to 40% faster than at the light intensity of 2500 Lux. It was concluded that the algae-bacteria consortia can effectively remove nitrogen from wastewater and the removal performance can be stabilized and enhanced using the low light intensity of 1000 Lux that is also a cost-effective strategy.

Eukaryotic Translesion DNA Synthesis on the Leading and Lagging Strands: Unique Detours around the Same Obstacle.

During S-phase, minor DNA damage may be overcome by DNA damage tolerance (DDT) pathways that bypass such obstacles, postponing repair of the offending damage to complete the cell cycle and maintain cell survival. In translesion DNA synthesis (TLS), specialized DNA polymerases replicate the damaged DNA, allowing stringent DNA synthesis by a replicative polymerase to resume beyond the offending damage. Dysregulation of this DDT pathway in human cells leads to increased mutation rates that may contribute to the onset of cancer. Furthermore, TLS affords human cancer cells the ability to counteract chemotherapeutic agents that elicit cell death by damaging DNA in actively replicating cells. Currently, it is unclear how this critical pathway unfolds, in particular, where and when TLS occurs on each template strand. Given the semidiscontinuous nature of DNA replication, it is likely that TLS on the leading and lagging strand templates is unique for each strand. Since the discovery of DDT in the late 1960s, most studies on TLS in eukaryotes have focused on DNA lesions resulting from ultraviolet (UV) radiation exposure. In this review, we revisit these and other related studies to dissect the step-by-step intricacies of this complex process, provide our current understanding of TLS on leading and lagging strand templates, and propose testable hypotheses to gain further insights.

Algal light sensing and photoacclimation in aquatic environments.

Anoxygenic photosynthetic prokaryotes arose in ancient oceans ~3.5 billion years ago. The evolution of oxygenic photosynthesis by cyanobacteria followed soon after, enabling eukaryogenesis and the evolution of complex life. The Archaeplastida lineage dates back ~1.5 billion years to the domestication of a cyanobacterium. Eukaryotic algae have subsequently radiated throughout oceanic/freshwater/terrestrial environments, adopting distinctive morphological and developmental strategies for adaptation to diverse light environments. Descendants of the ancestral photosynthetic alga remain challenged by a typical diurnally fluctuating light supply ranging from ~0 to ~2000 µE m-2  s-1 . Such extreme changes in light intensity and variations in light quality have driven the evolution of novel photoreceptors, light-harvesting complexes and photoprotective mechanisms in photosynthetic eukaryotes. This minireview focuses on algal light sensors, highlighting the unexpected roles for linear tetrapyrroles (bilins) in the maintenance of functional chloroplasts in chlorophytes, sister species to streptophyte algae and land plants.

Survival, DNA Integrity, and Ultrastructural Damage in Antarctic Cryptoendolithic Eukaryotic Microorganisms Exposed to Ionizing Radiation.

Life dispersal between planets, planetary protection, and the search for biosignatures are main topics in astrobiology. Under the umbrella of the STARLIFE project, three Antarctic endolithic microorganisms, the melanized fungus Cryomyces antarcticus CCFEE 515, a hyaline strain of Umbilicaria sp. (CCFEE 6113, lichenized fungus), and a Stichococcus sp. strain (C45A, green alga), were exposed to high doses of space-relevant gamma radiation (60Co), up to 117.07 kGy. After irradiation survival, DNA integrity and ultrastructural damage were tested. The first was assessed by clonogenic test; viability and dose responses were reasonably described by the linear-quadratic formalism. DNA integrity was evaluated by PCR, and ultrastructural damage was observed by transmission electron microscopy. The most resistant among the tested organisms was C. antarcticus both in terms of colony formation and DNA preservation. Besides, results clearly demonstrate that DNA was well detectable in all the tested organisms even when microorganisms were dead. This high resistance provides support for the use of DNA as a possible biosignature during the next exploration campaigns. Implication in planetary protection and contamination during long-term space travel are put forward. Key Words: Biosignatures-Ionizing radiation-DNA integrity-Eukaryotic microorganisms-Fingerprinting-Mars exploration. Astrobiology 17, 126-135.

Stress and Protists: No life without stress.

We report a summary of the symposium "Stress and Protists: No life without stress", which was held in September 2015 on the VII European Congress of Protistology in partnership with the International Society of Protistologists (Seville, Spain). We present an overview on general comments and concepts on cellular stress which can be also applied to any protist. Generally, various environmental stressors may induce similar cell responses in very different protists. Two main topics are reported in this manuscript: (i) metallic nanoparticles as environmental pollutants and stressors for aquatic protists, and (ii) ultraviolet radiation - induced stress and photoprotective strategies in ciliates. Model protists such as Chlamydomonas reinhardtii and Tetrahymena thermophila were used to assess stress caused by nanoparticles while stress caused by ultraviolet radiation was tested with free living planktonic ciliates as well as with the symbiont-bearing model ciliate Paramecium bursaria. For future studies, we suggest more intensive analyses on protist stress responses to specific environmental abiotic and/or biotic stressors at molecular and genetic levels up to ecological consequences and food web dynamics.

Gene expression characterizes different nutritional strategies among three mixotrophic protists.

Mixotrophic protists, i.e. protists that can carry out both phototrophy and heterotrophy, are a group of organisms with a wide range of nutritional strategies. The ecological and biogeochemical importance of these species has recently been recognized. In this study, we investigated and compared the gene expression of three mixotrophic protists, Prymnesium parvum, Dinobyron sp. and Ochromonas sp. under light and dark conditions in the presence of prey using RNA-Seq. Gene expression of the obligately phototrophic P. parvum and Dinobryon sp. changed significantly between light and dark treatments, while that of primarily heterotrophic Ochromonas sp. was largely unchanged. Gene expression of P. parvum and Dinobryon sp. shared many similarities, especially in the expression patterns of genes related to reproduction. However, key genes involved in central carbon metabolism and phagotrophy had different expression patterns between these two species, suggesting differences in prey consumption and heterotrophic nutrition in the dark. Transcriptomic data also offered clues to other physiological traits of these organisms such as preference of nitrogen sources and photo-oxidative stress. These results provide potential target genes for further exploration of the mechanisms of mixotrophic physiology and demonstrate the potential usefulness of molecular approaches in characterizing the nutritional modes of mixotrophic protists.

Repair of UV induced DNA lesions in ribosomal gene chromatin and the role of "Odd" RNA polymerases (I and III).

In fast growing eukaryotic cells, a subset of rRNA genes are transcribed at very high rates by RNA polymerase I (RNAPI). Nuclease digestion-assays and psoralen crosslinking have shown that they are open; that is, largely devoid of nucleosomes. In the yeast Saccharomyces cerevisae, nucleotide excision repair (NER) and photolyase remove UV photoproducts faster from open rRNA genes than from closed and nucleosome-loaded inactive rRNA genes. After UV irradiation, rRNA transcription declines because RNAPI halt at UV photoproducts and are then displaced from the transcribed strand. When the DNA lesion is quickly recognized by NER, it is the sub-pathway transcription-coupled TC-NER that removes the UV photoproduct. If dislodged RNAPI are replaced by nucleosomes before NER recognizes the lesion, then it is the sub-pathway global genome GG-NER that removes the UV photoproducts from the transcribed strand. Also, GG-NER maneuvers in the non-transcribed strand of open genes and in both strands of closed rRNA genes. After repair, transcription resumes and elongating RNAPI reopen the rRNA gene. In higher eukaryotes, NER in rRNA genes is inefficient and there is no evidence for TC-NER. Moreover, TC-NER does not occur in RNA polymerase III transcribed genes of both, yeast and human fibroblast.

Physiological aspects of UV-excitation of DNA.

Solar ultraviolet (UV) radiation, mainly UV-B (280-315 nm), is one of the most potent genotoxic agents that adversely affects living organisms by altering their genomic stability. DNA through its nucleobases has absorption maxima in the UV region and is therefore the main target of the deleterious radiation. The main biological relevance of UV radiation lies in the formation of several cytotoxic and mutagenic DNA lesions such as cyclobutane pyrimidine dimers (CPDs), 6-4 photoproducts (6-4PPs), and their Dewar valence isomers (DEWs), as well as DNA strand breaks. However, to counteract these DNA lesions, organisms have developed a number of highly conserved repair mechanisms such as photoreactivation, excision repair, and mismatch repair (MMR). Photoreactivation involving the enzyme photolyase is the most frequently used repair mechanism in a number of organisms. Excision repair can be classified as base excision repair (BER) and nucleotide excision repair (NER) involving a number of glycosylases and polymerases, respectively. In addition to this, double-strand break repair, SOS response, cell-cycle checkpoints, and programmed cell death (apoptosis) are also operative in various organisms to ensure genomic stability. This review concentrates on the UV-induced DNA damage and the associated repair mechanisms as well as various damage detection methods.

Potential impact of biofouling on the photobioreactors of the Offshore Membrane Enclosures for Growing Algae (OMEGA) system.

The influence of PBR composition [clear polyurethane (PolyU) vs. clear linear low-density polyethylene (LLDPE) (top) and black opaque high-density polyethylene (bottom)] and shape (rectangular vs. tubular) on biofouling and the influence of biofouling on algae productivity were investigated. In 9-week experiments, PBR biofouling was dominated by pennate diatoms and clear plastics developed macroalgae. LLDPE exhibited lower photosynthetic-active-radiation (PAR) light transmittance than PolyU before biofouling, but higher transmittance afterwards. Both rectangular and tubular LLDPE PBRs accumulated biofouling predominantly along their wetted edges. For a tubular LLDPE PBR after 12 weeks of biofouling, the correlation between biomass, percent surface coverage, and PAR transmittance was complex, but in general biomass inversely correlated with transmittance. Wrapping segments of this biofouled LLDPE around an algae culture reduced CO2 and NH3-N utilization, indicating that external biofouling must be controlled.

Does the Cyanophora paradoxa genome revise our view on the evolution of photorespiratory enzymes?

In the present-day O2 -rich atmosphere, the photorespiratory pathway is essential for organisms performing oxygenic photosynthesis; i.e. cyanobacteria, algae and land plants. The presence of enzymes for the plant-like 2-phosphoglycolate cycle in cyanobacteria indicates that, together with oxygenic photosynthesis, genes for photorespiratory enzymes were endosymbiotically conveyed from ancient cyanobacteria to photosynthetic eukaryotes. The genome information for Cyanophora paradoxa, a member of the Glaucophyta representing the first branching group of primary endosymbionts, and for many other eukaryotic algae was used to shed light on the evolutionary relationship of photorespiratory enzymes among oxygenic phototrophs. For example, it became possible to analyse the phylogenies of 2-phosphoglycolate phosphatase, serine:glyoxylate aminotransferase and hydroxypyruvate reductase. Analysis of the Cyanophora genome provided clear evidence that some photorespiratory enzymes originally acquired from cyanobacteria were lost, e.g. glycerate 3-kinase, while others were replaced by the corresponding enzymes from the α-proteobacterial endosymbiont, e.g. serine:glyoxylate aminotransferase. Generally, our analysis supports the view that many C2 cycle enzymes in eukaryotic phototrophs were obtained from the cyanobacterial endosymbiont, but during the subsequent evolution of algae and land plants multiple losses and replacements occurred, which resulted in a reticulate provenance of photorespiratory enzymes with different origins in different cellular compartments.

Energy-efficient photobioreactor configuration for algal biomass production.

An internally illuminated photobioreactor (IIPBR) design is proposed for energy-efficient biomass production. Theoretical rationale of the IIPBR design and its advantages over the traditional bubble column photobioreactors (PBRs) are presented, followed by experimental results from prototype scale cultivation of freshwater and marine algal strains in an 18L IIPBR. Based on theoretical considerations, the proposed IIPBR design has the potential to support 160% higher biomass density and higher biomass productivity per unit energy input, B/E, than a bubble column PBR of equal incident area per unit culture volume. Experimental B/E values recorded in this study with fresh water algae and marine algae (1.42 and 0.37 gW(-1)d(-1), respectively) are at least twice as those reported in the literature for comparable species cultivated in bubble column and airlift PBRs.

Short-term responses of unicellular planktonic eukaryotes to increases in temperature and UVB radiation.

BACKGROUND: Small size eukaryotes play a fundamental role in the functioning of coastal ecosystems, however, the way in which these micro-organisms respond to combined effects of water temperature, UVB radiations (UVBR) and nutrient availability is still poorly investigated. RESULTS: We coupled molecular tools (18S rRNA gene sequencing and fingerprinting) with microscope-based identification and counting to experimentally investigate the short-term responses of small eukaryotes (<6 µm; from a coastal Mediterranean lagoon) to a warming treatment (+3°C) and UVB radiation increases (+20%) at two different nutrient levels. Interestingly, the increase in temperature resulted in higher pigmented eukaryotes abundances and in community structure changes clearly illustrated by molecular analyses. For most of the phylogenetic groups, some rearrangements occurred at the OTUs level even when their relative proportion (microscope counting) did not change significantly. Temperature explained almost 20% of the total variance of the small eukaryote community structure (while UVB explained only 8.4%). However, complex cumulative effects were detected. Some antagonistic or non additive effects were detected between temperature and nutrients, especially for Dinophyceae and Cryptophyceae. CONCLUSIONS: This multifactorial experiment highlights the potential impacts, over short time scales, of changing environmental factors on the structure of various functional groups like small primary producers, parasites and saprotrophs which, in response, can modify energy flow in the planktonic food webs.

[Incomplete membrane restoration and radiation ageing].

An suggestion is put forward according to which the incomplete restoration of membranes in irradiated brain cells can self-perpetuate, down regulate their activity and accelerate ageing.