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61.
We have examined permeation by Ca(2+) and Ba(2+), and block by Mg(2+), using whole-cell recordings from alpha1G T-type calcium channels stably expressed in HEK 293 cells. Without Mg(o)(2+), inward currents were comparable with Ca(2+) and Ba(2+). Surprisingly, three other results indicate that alpha1G is actually selective for Ca(2+) over Ba(2+). 1) Mg(2+) block is approximately 7-fold more potent with Ba(2+) than with Ca(2+). With near-physiological (1 mM) Mg(o)(2+), inward currents were approximately 3-fold larger with 2 mM Ca(2+) than with 2 mM Ba(2+). The stronger competition between Ca(2+) and Mg(2+) implies that Ca(2+) binds more tightly than Ba(2+). 2) Outward currents (carried by Na(+)) are blocked more strongly by Ca(2+) than by Ba(2+). 3) The reversal potential is more positive with Ca(2+) than with Ba(2+), thus P(Ca) > P(Ba). We conclude that alpha1G can distinguish Ca(2+) from Ba(2+), despite the similar inward currents in the absence of Mg(o)(2+). Our results can be explained by a 2-site, 3-barrier model if Ca(2+) enters the pore 2-fold more easily than Ba(2+) but exits the pore at a 2-fold lower rate. 相似文献
62.
Nestedness has been widely used to measure the structure of biological communities and occurs when species-poor sites contain subsets of species-rich ones. Here, we examine nested patterns across the macroinvertebrate assemblages of 91 ponds in Doñana National Park, Spain, and explore temporal variation of nestedness and species richness in 19 temporary ponds over 2 years with differing rainfall. Macroinvertebrate assemblages were significantly nested; both pond spatial arrangement and environmental variation being important in driving nested patterns. Despite the nested structure observed, a number of taxa and ponds deviate from this pattern (termed idiosyncratic), by occurring more frequently than expected in species-poor sites, or having assemblages dominated by species largely absent from species-rich sites. Aquatic adults of winged insects, capable of dispersal, were more highly nested than non-dispersing taxa and life-history stages. Idiosyncratic taxa were found in ponds spanning a wide range of hydroperiods, although nestedness was higher in more permanent waterbodies. Monthly sampling demonstrated a gradual increase of species richness and nestedness from pond filling to April–May, when the most temporary ponds started to dry. Although the degree of nestedness of individual pond assemblages varied from month to month, the overall degree of nestedness in the two study years was practically identical despite marked differences in hydroperiod. Our results suggest that differential colonization and environmental variation are key processes driving the nested structure of Doñana ponds, that macroinvertebrate assemblages change in a predictable manner each year in response to cycles of pond wetting and drying, and that connectivity and environmental variability maintain biodiversity in pond networks. 相似文献
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64.
Sofia Graça Aranha Alexandra Teodósio Vânia Baptista Karim Erzini Ester Dias 《Journal of fish biology》2023,102(3):655-668
Deep-water sharks are among the most vulnerable deep-water taxa because of their extremely conservative life-history strategies (i.e., late maturation, slow growth, and reproductive rates), yet little is known about their biology and ecology. Thus, this study aimed at investigating the trophic ecology of five deep-water shark species, the birdbeak dogfish (Deania calcea), the arrowhead (D. profundorum), the smooth lanternshark (Etmopterus pusillus), the blackmouth catshark (Galeus melastomus) and the knifetooth dogfish (Scymnodon ringens) sampled onboard a crustacean bottom-trawler off the south-west coast of Portugal. We combined carbon and nitrogen stable isotopes with RNA and DNA (RD) ratios to investigate the main groups of prey assimilated by these species and their nutritional condition, respectively. Stable isotopes revealed overall small interspecific variability in the contribution of different taxonomic groups to sharks' tissues, as well as in the origin of their prey. S. ringens presented higher δ15N and δ13C values than the other species, suggesting reliance on bathyal cephalopods, crustaceans and teleosts; the remaining species likely assimilated bathy-mesopelagic prey. The RD ratios indicated that most of the individuals had an overall adequate nutritional condition and had recently eaten. This information, combined with the fact that stable isotopes indicate that sharks assimilated prey from the local or nearby food webs (including commercially important shrimps), suggests a potential overlap between this fishing area and their foraging grounds, which requires further attention. 相似文献
65.
Eva Borràs Ester Cantó Meena Choi Luisa Maria Villar José Carlos álvarez-Cerme?o Cristina Chiva Xavier Montalban Olga Vitek Manuel Comabella Eduard Sabidó 《Molecular & cellular proteomics : MCP》2016,15(1):318-328
Multiple sclerosis is an inflammatory, demyelinating, and neurodegenerative disease of the central nervous system. In most patients, the disease initiates with an episode of neurological disturbance referred to as clinically isolated syndrome, but not all patients with this syndrome develop multiple sclerosis over time, and currently, there is no clinical test that can conclusively establish whether a patient with a clinically isolated syndrome will eventually develop clinically defined multiple sclerosis. Here, we took advantage of the capabilities of targeted mass spectrometry to establish a diagnostic molecular classifier with high sensitivity and specificity able to differentiate between clinically isolated syndrome patients with a high and a low risk of developing multiple sclerosis. Based on the combination of abundances of proteins chitinase 3-like 1 and ala-β-his-dipeptidase in cerebrospinal fluid, we built a statistical model able to assign to each patient a precise probability of conversion to clinically defined multiple sclerosis. Our results are of special relevance for patients affected by multiple sclerosis as early treatment can prevent brain damage and slow down the disease progression.Multiple sclerosis is an inflammatory, demyelinating, and neurodegenerative disease of the central nervous system, and although the etiology of the disease is not fully understood, it is probably caused by the interaction of a complex genetic architecture and environmental factors. Multiple sclerosis affects over 2 million people worldwide, and it is typically diagnosed between ages 20 and 40, thus making a significant impact on public health and its economy (1).In most patients, the disease initiates with an episode of neurological disturbance referred to as clinically isolated syndrome. However, not all patients with this syndrome develop multiple sclerosis over time (2), and currently, the magnetic resonance imaging (MRI) abnormalities and the presence of IgG oligoclonal bands in cerebrospinal fluid (CSF) are used as predictors for later conversion to clinically definite multiple sclerosis (CDMS)1 (3–5). Although such abnormalities are considered important factors that influence the likelihood of developing CDMS, there is currently no clinical test that can conclusively establish whether a patient with a clinically isolated syndrome will eventually develop CDMS.The lack of diagnostic and prognostic biomarkers is a common problem for many diseases lacking a complete etiology, which is the case for most neurological disorders related to the central nervous system such as Parkinson''s and Alzheimer''s diseases, schizophrenia, and multiple sclerosis. In the particular case of multiple sclerosis, early treatment of patients with a clinically isolated syndrome can prevent brain damage and slow down the disease progression (6). Therefore, the availability of a diagnostic test in the initial stages of the disease is not only desirable but also of extreme relevance to attenuate the degenerative effects of the disease.Biomarker validation has traditionally been dominated by enzyme linked immuno-sorbent assays (ELISA), but recent advances in proteomics techniques have enabled the measurement of a subset of selected proteins over a large dynamic concentration range in multiple samples. Targeted mass spectrometry has thus become the method of choice when quantifying simultaneously a panel of proteins across many different biological samples (7–9). In particular, selected reaction monitoring (SRM) is the gold standard targeted mass spectrometry method for protein quantification due to its high precision, reliability, and throughput (10–13). This targeted mass spectrometry method is performed on triple quadrupole instruments, in which a predefined peptide precursor ion is first isolated, and then selected fragment ions arising from its collisional dissociation are measured over time. Each pair of precursor and fragment ion is called a transition, and multiple transitions can be coordinately measured and used to conclusively identify and quantify a peptide in a clinical complex sample.In a previous study, we used a screening mass spectrometric approach to discover potential markers for multiple sclerosis conversion in patients that initially presented a clinical isolated syndrome (14). In that discovery phase, quantitative mass spectrometry with iTRAQ labeling was used to measure protein abundances in pooled CSF samples from patients presenting a clinical isolated syndrome that either remained normal (CIS) or had eventually converted to clinically definite multiple sclerosis (CDMS) (n = 60). In the initial screening, several proteins exhibited significant differences in abundance when comparing these two groups of patients. The abundance change in one of the altered proteins, chitinase 3-like 1 (CH3L1), was confirmed by ELISA in CSF of individual patients, whereas for others, such as semaphorin 7A (SEM7A) and ala-β-his-dipeptidase (CNDP1), their abundance changes were confirmed by targeted mass spectrometry in follow-up studies with independent cohorts (15). Moreover, the levels of CH3L1 were associated with brain MRI abnormalities and disability progression during the follow-up period, as well as with shorter time to conversion to clinically definite multiple sclerosis (14).We now set out to establish a diagnostic protein classifier with high sensitivity and specificity able to differentiate between patients with a clinically isolated syndrome that have either a high or a low risk of developing clinically definite multiple sclerosis over time. For this purpose, CSF samples from an independent patient cohort from the one used in the discovery study were collected, and a set of preselected protein biomarker candidates were systematically quantified by targeted mass spectrometry (SRM) and evaluated for their classification power. Out of this study, we established a protein classifier based on the combination of abundances of proteins chitinase 3-like 1 and ala-β-his-dipeptidase, which is able to differentiate with high sensitivity and specificity between patients with a clinically isolated syndrome that have either a high or low risk of developing clinically definite multiple sclerosis. Moreover, the statistical model built around this protein classifier enables clinicians to easily assign to each patient a precise probability of conversion to clinically definite multiple sclerosis (Fig. 1).Open in a separate windowFig. 1.General workflow used in the present study. Initially, protein candidates identified in our previous discovery studies—together with several proteins described by other groups—were selected and quantified by targeted mass spectrometry (SRM) in a relatively large cohort individual patients. Protein quantities were then evaluated by their capability of classifying patients with clinical isolated syndrome, and thus, the best prognostic protein combination was identified. 相似文献
66.
Magda Vila Maria Grazia Giacobbe Mercedes Mas Ester Gangemi Antonella Penna Nagore Sampedro Filippo Azzaro Jordi Camp Luca Galluzzi 《Harmful algae》2005,4(4):673-695
Alexandrium minutum is a toxic dinoflagellate widespread along the Mediterranean coasts. This species is frequently detected year-round at low concentrations within the Mediterranean basin. However, it only proliferates recurrently in some localities. Two affected areas are the Catalan and Sicilian coasts. In order to identify the factors determining the A. minutum blooms in the Mediterranean Sea, we compare the bloom conditions in two harbours: Arenys de Mar (Catalan coast, Spain) and Syracuse (Sicily, Italy), during 2002–2003. Arenys de Mar harbour is a fishing and leisure harbour and receives an input of freshwater rich in nutrients. Likewise, the Syracuse harbour – located on the Ionian coast of Sicily – is subject to freshwater inputs. Some points of this site are used for productive activities such as shellfish farming. A. minutum from the two areas studied were morphologically and genetically identical. In both sites, recurrent blooms take place from winter to spring. Surface water temperatures and salinities during A. minutum bloom events were 12–14.5 °C and 32–38, and 16–24 °C and 32–37.7 for Arenys and Syracuse, respectively. During the blooms, the spatial distribution of A. minutum in the two harbours, the physicochemical characteristics and the phytoplankton community were studied. Similarities in composition of the phytoplankton community were evidenced, with a clear dominance of dinoflagellates over the other taxa. In Arenys, the second dominant species was Prorocentrum micans followed by Scrippsiella spp. and Dinophysis sacculus. The same species were found in Syracuse although P. triestinum, and alternatively Lingulodinium polyedrum, reached cell densities much higher than the other dinoflagellates giving marked water discolourations. 相似文献
67.
68.
Cerdà-Costa N Esteras-Chopo A Avilés FX Serrano L Villegas V 《Journal of molecular biology》2007,366(4):1351-1363
Understanding the initial steps of protein aggregation leading to the formation of amyloid fibrils remains a challenge. Here, the kinetics of such a process is determined for a misfolding protein model, ADA2h. The double nature of the very early kinetics suggests a step model of aggregation, where the denatured polypeptide folds into an aggregated beta-intermediate that subsequently reorganises into a more organised beta-sheet-richer structure that finally results in amyloid fibre formation. To determine the regions of the protein involved in amyloidosis, we have analysed a series of mutants previously made to study ADA2h folding. Using the algorithm TANGO, we have designed mutants that should enhance or decrease aggregation. Experimental analysis of the mutants shows that the C terminus of the molecule (comprising the last and edge beta-strand) is the major contributor to amyloid fibril formation, in good agreement with theoretical predictions. Comparison with proteins with similar topology reveals that family folds do not necessarily share the same principles of protein folding and/or aggregation. 相似文献
69.
Ferrer B Bermúdez-Silva FJ Bilbao A Alvarez-Jaimes L Sanchez-Vera I Giuffrida A Serrano A Baixeras E Khaturia S Navarro M Parsons LH Piomelli D Rodríguez de Fonseca F 《The Biochemical journal》2007,408(1):97-104
FgFtr1 and FgFtr2 are putative iron permeases, and FgFet1 and FgFet2 are putative ferroxidases of Fusarium graminearum. They have high homologies with iron permease ScFtr1 and ferroxidase ScFet3 of Saccharomyces cerevisiae at the amino acid level. The genes encoding iron permease and ferroxidase were localized to the same chromosome in the manner of FgFtr1/FgFet1 and FgFtr2/FgFet2. The GFP (green fluorescent protein)-fused versions of FgFtr1 and FgFtr2 showed normal functions when compared with FgFtr1 and FgFtr2 in an S. cerevisiae system, and the cellular localizations of FgFtr1 and FgFtr2 in S. cerevisiae depended on the expression of their putative ferroxidase partners FgFet1 and FgFet2 respectively. Although FgFtr1 was found on the plasma membrane when FgFet1 and FgFtr1 were co-transformed in S. cerevisiae, most of the FgFtr1 was found in the endoplasmic reticulum compartment when co-expressed with FgFet2. Furthermore, FgFtr2 was found on the vacuolar membrane when FgFet2 was co-expressed. From the two-hybrid analysis, we confirmed the interaction of FgFtr1 and FgFet1, and the same result was found between FgFtr2 and FgFet2. Iron-uptake activity also depended on the existence of the respective partner. Finally, the FgFtr1 and FgFtr2 were found on the plasma and vacuolar membrane respectively, in F. graminearum. Taken together, these results strongly suggest that FgFtr1 and FgFtr2 from F. graminearum encode the iron permeases of the plasma membrane and vacuolar membrane respectively, and require their specific ferroxidases to carry out normal function. Furthermore, the present study suggests that the reductive iron-uptake system is conserved from yeast to filamentous fungi. 相似文献
70.
López-Sánchez LM Collado JA Corrales FJ López-Cillero P Montero JL Fraga E Serrano J De La Mata M Muntané J Rodríguez-Ariza A 《Free radical research》2007,41(1):50-61
Nitric oxide (NO) participates in the cell death induced by d-Galactosamine (d-GalN) in hepatocytes, and NO-derived reactive oxygen intermediates are critical contributors to protein modification and hepatocellular injury. It is anticipated that S-nitrosation of proteins will participate in the mechanisms leading to cell death in d-GalN-treated human hepatocytes. In the present study, d-GalN-induced cell death was related to augmented levels of NO production and S-nitrosothiol (SNO) content. The biotin switch assay confirmed that d-GalN increased the levels of S-nitrosated proteins in human hepatocytes. S-nitrosocysteine (CSNO) enhanced protein S-nitrosation and altered cell death parameters that were related to S-nitrosation of the executioner caspase-3. Fifteen S-nitrosated proteins participating in metabolism, antioxidative defense and cellular homeostasis were identified in human hepatocytes treated with CSNO. Among them, seven were also identified in d-GalN-treated hepatocytes. The results here reported underline the importance of the alteration of SNO homeostasis during d-GalN-induced cell death in human hepatocytes. 相似文献