Ca2+ activates glycogenolysis in isolated mantle storage cells of Mytilus galloprovincialis Lmk.

Glycogenolytic activity (GA) in isolated mantle storage cells (MSC) from Mytilus galloprovincialis was studied, while glycogen and free-glucose content, as well as glucose released from cells were tested. In the period studied (November-December), the glucose releasing activity measured can be considered as an output of GA. In both, whole cells system (WCS) and crude cell-free system (CFS), a non-stimulated GA was detected. In WCS, dopamine and 5-hydroxytryptamine (5-HT) stimulated glycogenolysis, while epinephrine, norepinephrine and isoproterenol did not show any effect. Furthermore, mellitin and the Ca(2+)-ionophore, A23187, had a stimulating effect on the GA. In CFS, the absence of Ca2+ ions was a sufficient condition to depress GA. These and other findings suggest that: 1) GA in MSC may be stimulated by dopamine and 5-HT and not by adrenergic agonists; 2) cytosolyc Ca2+ signalling may have become an absolute requirement for activation of the glycogenolytic cascade in MSC; 3) a rapid high-affinity glucose transport may occur in these cells.     

EDC3 phosphorylation regulates growth and invasion through controlling P‐body formation and dynamics

Regulation of mRNA stability and translation plays a critical role in determining protein abundance within cells. Processing bodies (P‐bodies) are critical regulators of these processes. Here, we report that the Pim1 and 3 protein kinases bind to the P‐body protein enhancer of mRNA decapping 3 (EDC3) and phosphorylate EDC3 on serine (S)161, thereby modifying P‐body assembly. EDC3 phosphorylation is highly elevated in many tumor types, is reduced upon treatment of cells with kinase inhibitors, and blocks the localization of EDC3 to P‐bodies. Prostate cancer cells harboring an EDC3 S161A mutation show markedly decreased growth, migration, and invasion in tissue culture and in xenograft models. Consistent with these phenotypic changes, the expression of integrin β1 and α6 mRNA and protein is reduced in these mutated cells. These results demonstrate that EDC3 phosphorylation regulates multiple cancer‐relevant functions and suggest that modulation of P‐body activity may represent a new paradigm for cancer treatment.     

Mutational analysis of Asp51 of Anabaena azollae glutamine synthetase. D51E mutation confers resistance to the active site inhibitors L-methionine-DL-sulfoximine and phosphinothricin.

The role of Asp51 in the catalytic activity of glutamine synthetase from the cyanobacterium Anabaena azollae has been analyzed. Five mutant enzymes, D51S, D51A, D51E, D51N and D51R, were constructed by site-directed mutagenesis and characterized. Asp51 appears to participate in the binding of ammonium ion, as affinity for this substrate was affected in all cases, although it varied according to the charge and/or size of the amino-acid residue, decreasing in the order Glu > Asn > Ser > Ala. The replacement of Asp51 by Glu (D51E) conferred besides a high resistance to the herbicides L-methionine-DL-sulfoximine and phosphinothricin, as a result of a decreased phosphorylation ability.     

Perspectives of fluorescence spectroscopy for online monitoring in microalgae industry

Microalgae industrial production is viewed as a solution for alternative production of nutraceuticals, cosmetics, biofertilizers, and biopolymers. Throughout the years, several technological advances have been implemented, increasing the competitiveness of microalgae industry. However, online monitoring and real-time process control of a microalgae production factory still require further development. In this mini-review, non-destructive tools for online monitoring of cellular agriculture applications are described. Still, the focus is on the use of fluorescence spectroscopy to monitor several parameters (cell concentration, pigments, and lipids) in the microalgae industry. The development presented makes it the most promising solution for monitoring up-and downstream processes, different biological parameters simultaneously, and different microalgae species. The improvements needed for industrial application of this technology are also discussed.     

Antibody-based diagnosis of small ruminant lentivirus infection in seminal fluid

Antibody-based diagnosis of small ruminant lentiviruses (SRLVs) has been efficiently achieved using serum and milk, but not semen, for which polymerase chain reaction (PCR) has been proposed as a confirmatory technique. This work, involving 296 ovine (Ovis aries) and caprine (Capra hircus) semen donors, investigates whether seminal fluid (SF) can be reliably used in antibody-based SRLV diagnosis. First, a gold standard was established to assess the infection status and determine the sensitivity and specificity of three commercial enzyme-linked immunosorbent assays (ELISAs) in serum testing using Western blot and PCR as confirmatory tests. For SF testing, both gold standard and serum testing results were used as reference. The performance of SF testing was affected not only by the ELISA assay sensitivity (related to antigen spectrum) compared with that of the gold standard (as it occurred in serum testing) but also by SF sample quality and SF working dilution. Nonturbid SF samples, commonly collected in artificial insemination centers (AICs), were required. Compared with serum, SF testing had a decreased sensitivity in two of the ELISA assays (with original serum working dilutions ≤1/20 in serum testing) but reached a similar sensitivity (and specificity) in the assay designed to work at the highest serum dilution (1/500). A SF concentration of about 1/2 (250-fold that used in serum testing) was found optimal in this assay, yielding highly repeatable results that were in almost perfect agreement with those of serum testing (κ ± SE, 0.91 ± 0.81). Thus, SF ELISA can be reliably applied in antibody-based SRLV diagnosis. This information may be useful to control infection in AICs and animal and semen trade programs requiring health-certified quality of semen donors.     

Coccomyxa sp. (Chlorophyta: Chlorococcales), a new pathogen in mussels (Mytilus galloprovincialis) of Vigo estuary (Galicia, NW Spain)

In this work, we describe the occurrence of irregular shaped green aggregations in the mantle, gill filaments, adductor muscle, visceral mass and haemolymph of wild mussels (Mytilus galloprovincialis) collected from the Vigo estuary (Galicia, NW Spain). Microscopic examination of these masses revealed that they consist of intracellular green algae which are spherical to oval in shape, 5 μm in length and 3 μm in width, without flagella and with a smooth surface. The algal cells present a small single nucleus, a mitochondrion, 1-2 parietal chloroplasts and lack pyrenoids. Reproduction is by formation of 2-4 autospores or daughter cells. Pigment analysis reveals the presence of photopigments typical of green algae in addition to alloxanthin, diadinoxanthin and diatoxanthin. These carotenoids are noted for the first time in a parasitic chlorophyte. The signs of infection, together with the morphological observations, suggest that this parasitic algae may be Coccomyxa parasitica. However, further molecular studies are required for confirmation. This is the first report of Coccomyxa algae parasitizing the species M. galloprovincialis.     

Stream friendly coffee: evaluating the impact of coffee farming on high-elevation streams of the Tarrazú coffee region of Costa Rica

Coffee farming is an important practice in the tropics, but studies addressing its effect on streams, especially at high elevations in high productivity regions, are limited. We evaluated streams draining the high-elevation coffee farming region of Tarrazú, Costa Rica, in terms of water quality and benthic macroinvertebrates over the course of a year. We compared our observations to regulatory benchmarks to determine overall condition, and compared our sites in terms of riparian vegetation (i.e., canopy cover, and riparian width) to determine the role of this management practice. Overall, we found that most streams fell within recommended water quality criteria and supported high proportion of pollution-sensitive taxa. Nevertheless, we found high levels of conductivity during the drier period of the year, which may signal agrochemical pollution. Also during the drier period, we found significantly higher diversity, % scrapers, and % predators, which suggests a boost in basal food sources associated to the observed high conductivity levels. We found that riparian buffer widths were not associated to improvements in water quality indicators or bio-integrity parameters on our sites. However, canopy cover was positively correlated to % shredders and family diversity, and negatively correlated to certain tolerant taxa. Our findings help to better characterize a system that is of great economic importance in the tropics, but that remains relatively under studied. It also helps inform management practices to enhance the potential of coffee agroforestry systems in promoting tropical biodiversity conservation within aquatic ecosystems.