Choline kinase inhibition induces exacerbated endoplasmic reticulum stress and triggers apoptosis via CHOP in cancer cells

Endoplasmic reticulum (ER) is a central organelle in eukaryotic cells that regulates protein synthesis and maturation. Perturbation of ER functions leads to ER stress, which has been previously associated with a broad variety of diseases. ER stress is generally regarded as compensatory, but prolonged ER stress has been involved in apoptosis induced by several cytotoxic agents. Choline kinase α (ChoKα), the first enzyme in the Kennedy pathway, is responsible for the generation of phosphorylcholine (PCho) that ultimately renders phosphatidylcholine. ChoKα overexpression and high PCho levels have been detected in several cancer types. Inhibition of ChoKα has demonstrated antiproliferative and antitumor properties; however, the mechanisms underlying these activities remain poorly understood. Here, we demonstrate that ChoKα inhibitors (ChoKIs), MN58b and RSM932A, induce cell death in cancer cells (T47D, MCF7, MDA-MB231, SW620 and H460), through the prolonged activation of ER stress response. Evidence of ChoKIs-induced ER stress includes enhanced production of glucose-regulated protein, 78 kDa (GRP78), protein disulfide isomerase, IRE1α, CHOP, CCAAT/enhancer-binding protein beta (C/EBPβ) and TRB3. Although partial reduction of ChoKα levels by small interfering RNA was not sufficient to increase the production of ER stress proteins, silencing of ChoKα levels also show a decrease in CHOP overproduction induced by ChoKIs, which suggests that ER stress induction is due to a change in ChoKα protein folding after binding to ChoKIs. Silencing of CHOP expression leads to a reduction in C/EBPβ, ATF3 and GRP78 protein levels and abrogates apoptosis in tumor cells after treatment with ChoKIs, suggesting that CHOP maintains ER stress responses and triggers the pro-apoptotic signal. Consistent with the differential effect of ChoKIs in cancer and primary cells previously described, ChoKIs only promoted a transient and moderated ER stress response in the non-tumorogenic cells MCF10A. In conclusion, pharmacological inhibition of ChoKα induces cancer cell death through a mechanism that involves the activation of exaggerated and persistent ER stress supported by CHOP overproduction.     

Sponge Mass Mortalities in a Warming Mediterranean Sea: Are Cyanobacteria-Harboring Species Worse Off?

Mass mortality events are increasing dramatically in all coastal marine environments. Determining the underlying causes of mass mortality events has proven difficult in the past because of the lack of prior quantitative data on populations and environmental variables. Four-year surveys of two shallow-water sponge species, Ircinia fasciculata and Sarcotragus spinosulum, were carried out in the western Mediterranean Sea. These surveys provided evidence of two severe sponge die-offs (total mortality ranging from 80 to 95% of specimens) occurring in the summers of 2008 and 2009. These events primarily affected I. fasciculata, which hosts both phototrophic and heterotrophic microsymbionts, while they did not affect S. spinosulum, which harbors only heterotrophic bacteria. We observed a significant positive correlation between the percentage of injured I. fasciculata specimens and exposure time to elevated temperature conditions in all populations, suggesting a key role of temperature in triggering mortality events. A comparative ultrastructural study of injured and healthy I. fasciculata specimens showed that cyanobacteria disappeared from injured specimens, which suggests that cyanobacterial decay could be involved in I. fasciculata mortality. A laboratory experiment confirmed that the cyanobacteria harbored by I. fasciculata displayed a significant reduction in photosynthetic efficiency in the highest temperature treatment. The sponge disease reported here led to a severe decrease in the abundance of the surveyed populations. It represents one of the most dramatic mass mortality events to date in the Mediterranean Sea.     

Evidence of negative impacts of ecological tourism on turtlegrass (<Emphasis Type="Italic">Thalassia testudinum</Emphasis>) beds in a marine protected area of the Mexican Caribbean

Many marine protected areas (MPAs) have been established in recent years. Some MPAs are open to tourists to foster environmental education and generate revenue for the MPA. This has been coined “ecological tourism”. Here, we examine the impact of ecological tourism on turtlegrass (Thalassia testudinum) health in one area of the “Costa Occidental de Isla Mujeres, Punta Cancún y Punta Nizuc” MPA in the Mexican Caribbean. A heavily visited location was compared with an unvisited location. Turtlegrass leaves at the visited location were sparser, shorter, grew more slowly, and had more epiphytes than at the unvisited location. Vertical and horizontal rhizomes of turtlegrass also grew more slowly at the visited than at the unvisited location. There is reasonable evidence to suggest that the observed differences are likely due to the deleterious impacts of novice and careless snorkelers. If continuing, these impacts could cause severe degradation of the visited areas in this MPA and, thus, changes in management policies seem in order.     

Coexistence of Low Coral Cover and High Fish Biomass at Farquhar Atoll,Seychelles

We report a reef ecosystem where corals may have lost their role as major reef engineering species but fish biomass and assemblage structure is comparable to unfished reefs elsewhere around the world. This scenario is based on an extensive assessment of the coral reefs of Farquhar Atoll, the most southern of the Seychelles Islands. Coral cover and overall benthic community condition at Farquhar was poor, likely due to a combination of limited habitat, localized upwelling, past coral bleaching, and cyclones. Farquhar Atoll harbors a relatively intact reef fish assemblage with very large biomass (3.2 t ha⁻¹) reflecting natural ecological processes that are not influenced by fishing or other local anthropogenic factors. The most striking feature of the reef fish assemblage is the dominance by large groupers, snappers, and jacks with large (>1 m) potato cod (Epinephelus tukula) and marbled grouper (E. polyphekadion), commonly observed at many locations. Napoleon wrasse (Cheilinus undulatus) and bumphead parrotfish (Bolbometopon muricatum) are listed as endangered and vulnerable, respectively, but were frequently encountered at Farquhar. The high abundance and large sizes of parrotfishes at Farquhar also appears to regulate macroalgal abundance and enhance the dominance of crustose corallines, which are a necessary condition for maintenance of healthy reef communities. Overall fish biomass and biomass of large predators at Farquhar are substantially higher than other areas within the Seychelles, and are some of the highest recorded in the Indian Ocean. Remote islands like Farquhar Atoll with low human populations and limited fishing pressure offer ideal opportunities for understanding whether reefs can be resilient from global threats if local threats are minimized.     

Role of first-order consumers in ecosystem carbon flow

Herbivores and detritus consumers (i.e. microbial decomposers and invertebrate and vertebrate detritivores) are pivotal components of trophic food webs and thus play a paramount role in the trophic transference and turnover of producer‐fixed carbon. Hence, elucidating patterns in carbon flux through these first‐order consumers is important to understand the nature and controls of carbon flow in ecosystems. Here, using the largest literature compilation to date, I show that, in contrast with the current belief, aquatic herbivores accumulate on average three times as much biomass as do terrestrial herbivores for a given level of primary production and, as a consequence, turn over the ingested carbon only slightly faster than do terrestrial herbivores. Conversely, aquatic detritus consumers generally accumulate a much lower biomass (i.e. over ten times lower) than their terrestrial counterparts for a given level of primary production and, thus, they turn over the ingested carbon much more quickly (i.e. over ten times faster). Because the detrital pathway generally dominates the trophic flow of carbon in both aquatic and terrestrial ecosystems, carbon also tends to flow through the total compartment of first order‐consumers (both herbivores and detritus consumers) at a much faster rate in aquatic than in terrestrial ecosystems. Thus, aquatic systems, because of faster carbon recycling rates through their basal and first‐order levels of the food chain, appear to have a lower capacity than do terrestrial systems for retaining carbon under natural or anthropogenic increases in photosynthetic fixation.     

Aggregation dynamics and foraging behaviour of striped red mullet Mullus surmuletus in the western Mediterranean Sea

Habitat‐related heterogeneity of striped red mullet Mullus surmuletus heterospecific foraging assemblages was examined off the coast of Spain. Video‐based focal‐follows conducted on 122 M. surmuletus assemblages (446 total individuals) revealed an array of attendant species (n = 7) with composition linked to benthic habitat complexity; bare sandy substrata were characterized by homospecific groups of M. surmuletus, while habitats with rock and vegetation attracted a variety of scrounging labrids and sparids. Although the nature of the relationship between M. surmuletus and attendants requires further exploration, the present study indicates that substratum composition can be a driving factor explaining the dynamics of this heterospecific assemblage.     

Genome structure of cottontail rabbit herpesvirus.

The genome structure of a herpesvirus isolated from primary cultures of kidney cells from the cottontail rabbit Sylvilagus floridanus was elucidated by using electron microscopy and restriction enzyme analysis. The genome, which was about 150 kilobase pairs long and which had an average G + C composition of 45%, consisted of two regions with unique base sequences (54 and 47 kilobase pairs) enclosed by reiterations of a 925-base-pair sequence with a variable copy number. The internal repeats were in opposite polarity with respect to the terminal repeats, and both unique regions underwent inversion. The nucleotide sequence of the repeat unit was determined, and virion DNA termini were precisely localized within this sequence. Elements showing homology with the cleavage-packaging signals common to other herpesviruses were detected. The data indicate that this virus is different from the previously described herpesvirus sylvilagus.     

"Endless" viral DNA in cells infected with channel catfish virus

The state of intracellular viral DNA in cells infected with channel catfish virus has been studied by the Hirt selective extraction procedure and by restriction endonuclease digestion. The sedimentation properties and restriction patterns of viral DNA in the Hirt supernatant fraction indicate that the majority, if not all, of the DNA is in the form of linear unit-length (Mr approximately equal to 85 x 10(6)) molecules. However, restriction digests of viral DNA in the pellet fraction lacked two fragments corresponding to the molecular ends of unit-length DNA. In addition, there appeared in HpaI digests of pellet DNA a new restriction fragment interpretable as the product of fusion between the ends of unit-length molecules. The size of the new fragment requires that fusion occur in such a way that one copy of the terminally repeated sequences (Mr approximately equal to 12.3 x 10(6)) of the unit-length DNA is lost in the process. In pulse-chase experiments, radioactivity flowed from the pellet fraction to the supernatant fraction, suggesting a precursor-product relationship for these DNA species. The results are easily understood if unit-length virion DNA is generated by excision from concatemeric structures.     

Effects of UV radiation and nitrate limitation on the production of biogenic sulfur compounds by marine phytoplankton

We tested the effects of UV radiation (UVR) and nitrate limitation on the production of dimethylsulfide (DMS), particulate dimethylsulfoniopropionate (DMSPp), and particulate dimethylsulfoxide (DMSOp) in natural seawater from the Gulf of Mexico and in phytoplankton cultures. DMS/Chl a ratios in PAR-only and PAR + UV-exposed seawater were 0.44–2.0 and 0.46–1.9 nmol DMS μg⁻¹ Chl a, respectively, whereas the ratios in cultures of Amphidinium carterae were 1.0–2.2 nmol μg⁻¹ in PAR-exposed samples and 0.91–2.1 nmol μg⁻¹ in PAR + UV-exposed samples. These results suggested that UVR did not substantially affect DMS/Chl a ratios in seawater and A. carterae culture samples. Similarly, UVR had no significant effect on DMSOp/Chl a in seawater samples (0.83–1.6 nmol DMSO μg⁻¹ Chl a for PAR + UV vs. 0.70–1.5 nmol μg⁻¹ for PAR-exposed seawater samples, respectively) or in A. carterae cultures (0.20–1.3 and 0.19–0.88 nmol DMSO μg⁻¹ Chl a in PAR + UV- and PAR-exposed cultures, respectively). In an experiment with the diatom, Thalassiosira oceanica, the culture was grown in high nitrate (30 μM) or low nitrate (6 μM) media and exposed to PAR-only or PAR + UV. The low nitrate, PAR-only samples showed an increase of intracellular dimethylsulfoniopropionate (DMSP) concentration from 2.1 to 15 mmol L⁻¹ in 60 h, but the increase occurred only after cultures reached the stationary phase. Cultures of T. oceanica grown under UVR had lower growth rates than those under PAR-only (μ′ = 0.17 and 0.32 d⁻¹, respectively) and perhaps did not experience severe nitrate limitation even in the low nitrate treatment. These results suggest that the elevated UVR in low nitrate environments could result in reduction of DMSP in some species, whereas DMSP concentrations would not be affected in eutrophic areas.