Empirical evidence for large‐scale human impact on intertidal aggregations,larval supply and recruitment of Pyura praeputialis around the Bay of Antofagasta,Chile

In northern Chile, Pyura praeputialis is an invasive species inhabiting rocky intertidal and subtidal habitats restricted exclusively to the Bay of Antofagasta where it forms extensive aggregations. The negative impact of Pyura gathering on mid‐intertidal abundances of this species has recently been reported at the south‐eastern end of this bay. In the present study we have increased sampling sites to cover the entire bay toward the north‐western end and the northern section, where a coastal marine reserve for the scallop fishery partially restricts shellfish gathering. Therefore, the sampling sites were chosen to represent different levels of shellfish gathering access along the northern shore of the bay. Long‐term monitoring (1999–2014) of changes in tunicate cover and the abundances of larvae and recruits at seven sites are reported. The opening of a remodelled artificial and recreational beach in 2012, on the central‐eastern shore of the bay, has increased accessibility to rocky intertidal platforms that started to be massively visited by Pyura gatherers from the summer of 2013. This allowed for the implementation of an intensive short‐term monitoring program of changes in tunicate cover and the abundances of their larvae and recruits. When gathering access was present the reduction in intertidal cover was generalized to the entire bay and followed by reductions in larvae and recruits. However, these reductions were not found in sites with more restricted gathering access. We conclude that continuous extraction by Pyura gatherers followed by reductions of conspecific larvae and recruits are the main drivers behind the reduced abundance of P. praeputialis in the entire bay of Antofagasta. Thus, if gathering is not stopped important ecosystem services provided by this tunicate in the bay may be threatened. Similar consequences may be expected if other massive and irreversible reductions in other species of the Pyura complex, that inhabit other coasts in the southern hemisphere, occur. The controversy concerning the impacts of invasive species and whether they cause negative, positive or neutral impacts to original ecosystems and fisheries is discussed.     

Separate taurine and chloride efflux pathways activated during regulatory volume decrease

Organic osmolyte and halide permeability pathways activated inepithelial HeLa cells by cell swelling were studied by radiotracer efflux techniques and single-cell volume measurements. The replacement of extracellular Cl byanions that are more permeant through the volume-activated Cl channel, as indicated byelectrophysiological measurements, significantly decreasedtaurine efflux. In the presence of less-permeant anions, an increase intaurine efflux was observed. Simultaneous measurement of the¹²⁵I, used as a tracer forCl, and[³H]taurine effluxshowed that the time courses for the two effluxes differed. InCl-rich medium the increasein I efflux was transient,whereas that for taurine was sustained. OsmosensitiveCl conductance, assessed bymeasuring changes in cell volume, increased rapidly after hypotonicshock. The influx of taurine was able to counteractCl conductance-dependentcell shrinkage but only ~4 min after triggering cell swelling. Thistaurine-induced effect was blocked by DIDS. Differences in anionsensitivity, the time course of activation, and sensitivity to DIDSsuggest that the main cell swelling-activated permeability pathways fortaurine and Cl are separate.

     

Kin or self‐recognition? Colonial fusibility of the bryozoan Celleporella hyalina

We estimated fusion frequency with respect to coancestry in the bryozoan Celleporella hyalina, whose briefly planktonic sexually produced larvae settle on algal substrata and proceed to form encrusting colonies by iterative budding. Frequency of fusion between paired colonies growing on an artificial substratum was positively correlated with coefficient of relatedness, with allorecognition ability increasing during the early stages of colonial growth after larval settlement. Parents repressed the growth of F1 progeny with which they had fused. The results are concordant with the Feldgarden-Yund model of selection for self-recognition, which regards fusion with kin as an inevitable source of error whose cost diminishes with increasing relatedness. Contrary to fusion compatibility, gametic compatibility is negatively correlated with coancestry, indicating a need for further research on the possibility of common or linked genetic control that has opposite effect at somatic and gametic levels.     

Revisiting the mouse mitochondrial DNA sequence

The existence of reliable mtDNA reference sequences for each species is of great relevance in a variety of fields, from phylogenetic and population genetics studies to pathogenetic determination of mtDNA variants in humans or in animal models of mtDNA-linked diseases. We present compelling evidence for the existence of sequencing errors on the current mouse mtDNA reference sequence. This includes the deletion of a full codon in two genes, the substitution of one amino acid on five occasions and also the involvement of tRNA and rRNA genes. The conclusions are supported by: (i) the re-sequencing of the original cell line used by Bibb and Clayton, the LA9 cell line, (ii) the sequencing of a second L-derivative clone (L929), and (iii) the comparison with 12 other mtDNA sequences from live mice, 10 of them maternally related with the mouse from which the L cells were generated. Two of the latest sequences are reported for the first time in this study (Balb/cJ and C57BL/6J). In addition, we found that both the LA9 and L929 mtDNAs also contain private clone polymorphic variants that, at least in the case of L929, promote functional impairment of the oxidative phosphorylation system. Conse quently, the mtDNA of the strain used for the mouse genome project (C57BL/6J) is proposed as the new standard for the mouse mtDNA sequence.     

Nitric oxide triggers the toxicity due to glutathione depletion in midbrain cultures through 12-lipoxygenase

Glutathione (GSH) depletion is the earliest biochemical alteration shown to date in brains of Parkinson's disease patients. However, data from animal models show that GSH depletion by itself is not sufficient to induce nigral degeneration. We have previously shown that non-toxic inhibition of GSH synthesis with l-buthionine-(S,R)-sulfoximine in primary midbrain cultures transforms a nitric oxide (NO) neurotrophic effect, selective for dopamine neurons, into a toxic effect with participation of guanylate cyclase (GC) and cGMP-dependent protein kinase (PKG) (Canals, S., Casarejos, M. J., de Bernardo, S., Rodríguez-Martín, E., and Mena, M. A. (2001) J. Neurochem. 79, 1183-1195). Here we demonstrate that arachidonic acid (AA) metabolism through the 12-lipoxygenase (12-LOX) pathway is also central for this GSH-NO interaction. LOX inhibitors (nordihydroguaiaretic acid and baicalein), but not cyclooxygenase (indomethacin) or epoxygenase (clotrimazole) ones, prevent cell death in the culture, even when added 10 h after NO treatment. Furthermore, the addition of AA to GSH-depleted cultures precipitates a cell death process that is indistinguishable from that initiated by NO in its morphology, time course, and 12-LOX, GC, and PKG dependence. The first AA metabolite through the 12-LOX enzyme, 12-hydroperoxyeicosatetraenoic acid, induces cell death in the culture, and its toxicity is greatly enhanced by GSH depletion. In addition we show that if GSH synthesis inhibition persists for up to 4 days without any additional treatment, it will induce a cell death process that also depends on 12-LOX, GC, and PKG activation. In this study, therefore, we show that the signaling pathway AA/12-LOX/12-HPETE/GC/PKG may be important in several pathologies in which GSH decrease has been documented, such as Parkinson's disease. The potentiating effect of NO over such a signaling pathway may be of relevance as part of the cascade of events leading to and sustaining nerve cell death.     

Ligand-dependent structural changes and limited proteolysis of Escherichia coli phosphofructokinase-2

An isoform (rhesus UGT1A01) orthologus to the human UGT1A1 was cloned and sequenced from female rhesus monkey liver cDNA using primers designed from the human nucleotide sequences. Open reading frame analysis of the PCR-generated product encodes a 533-amino acid protein with a proposed 27-residue signal peptide. Nucleotide sequence comparison of rhesus UGT1A01 to other rhesus UGT1A isoforms detected a single-transition mutation at nucleotide 1520 (T-->C), resulting in a neutral F to S substitution at position 507. Rhesus UGT1A01 was greater than 99 and 95% identical to cynomolgus UGT1A01 and human UGT1A1, respectively. The rhesus UGT1A01 was expressed in HK-293 cells for functional analysis. Catalytic activity of UGT1A01 was determined with 7-hydroxy-4-(trifluoromethyl)-coumarin and more specific human UGT1A1 substrates (1-naphthol, beta-estradiol, 17 alpha-ethinylestradiol, and bilirubin). Expression of UGT1A01 protein was also detected by a Western blot utilizing a polyclonal antibody developed against the human UGT1A family.     

The role of astroglia on the survival of dopamine neurons

Glial cells play a key role in the function of dopamine (DA) neurons and regulate their differentiation, morphology, physiological and pharmacological properties, survival, and resistance to different models of DA lesion. Several studies suggest that glial cells may be important in the pathogenesis of Parkinson’s disease (PD), a common neurodegenerative disorder characterized by degeneration of the nigrostriatal DA system. In this disease the role of glia could be due to the excessive production of toxic products such as nitric oxide (NO) or cytokines characteristic of inflammatory process, or related to a defective release of neuroprotective agents, such as small antioxidants with free radical scavenging properties or peptidic neurotrophic factors.     

Absolute TCD4+ counting by a minimalist dual-platform flow cytometric method

The aim of this work was to compare the performance of an absolute TCD4+ counting method based on total WBC gating versus the standard lymphocyte (Ly) gating method, in order to develop a flow cytometric (FCM) minimalist strategy for TCD4+ enumeration. METHOD: 132 routine peripheral blood samples, mainly from HIV infected patients, were labelled with CD3-FITC/CD4-PE/CD45-PECy5 and analyzed by two gating methods: a) standard method based on Ly immunological gating (CD45++SSClow), followed by the determination of CD3+CD4+ percentage and absolute number (# calculation using Ly # from hematological analyser (HA); b) total WBC immunological gate on biparametric scatter CD45/CD4, followed by CD4++SSClow percentage determination and absolute number calculation using WBC absolute number from hematological counter without using the WBC differential. Moreover on 63 samples Ly # based on Ly % from FCM and WBC counting from HA was compared with Ly # from HA. RESULTS: The TCD4+/microL ranged from 3 to 3277 and the statistical analysis results showed: a) linear regression: r2 = 0.9847; b) Bland & Altman analysis: difference mean = -56.22; agreement range = +95.68 / -208.12; c) the mean of result difference/mean value*100 between two methods was -9.06%; d) comparison between regression line and the boundaries for acceptable residual values based on regressed confidence limits found by A. Kunkl et al showed regression line within boundaries near the upper limits. The Ly/microL count ranged from 635 to 8752. The statistical analysis results showed: a) linear regression: r2 = 0.9764; b) Bland & Altman analysis: difference mean = -362.93; agreement range = +134.51 / -860.37; c) the mean of result difference/mean value*100 between two methods was -16.12%. CONCLUSIONS: Our results suggest a fair agreement between the two gating methods, but the one based on total WBC gate gives TCD4+/microL counts systematically higher than the standard method. This finding can be attributed to a systematic lower estimation of Ly% by HA.