Centrioles generate a local pulse of Polo/PLK1 activity to initiate mitotic centrosome assembly.

Mitotic centrosomes are formed when centrioles start to recruit large amounts of pericentriolar material (PCM) around themselves in preparation for mitosis. This centrosome "maturation" requires the centrioles and also Polo/PLK1 protein kinase. The PCM comprises several hundred proteins and, in Drosophila, Polo cooperates with the conserved centrosome proteins Spd-2/CEP192 and Cnn/CDK5RAP2 to assemble a PCM scaffold around the mother centriole that then recruits other PCM client proteins. We show here that in Drosophila syncytial blastoderm embryos, centrosomal Polo levels rise and fall during the assembly process-peaking, and then starting to decline, even as levels of the PCM scaffold continue to rise and plateau. Experiments and mathematical modelling indicate that a centriolar pulse of Polo activity, potentially generated by the interaction between Polo and its centriole receptor Ana1 (CEP295 in humans), could explain these unexpected scaffold assembly dynamics. We propose that centrioles generate a local pulse of Polo activity prior to mitotic entry to initiate centrosome maturation, explaining why centrioles and Polo/PLK1 are normally essential for this process.

Evolution of the ecological niche behind the largest disjunct freshwater fish distribution in the world.

Ecological processes that are behind distributions of species that inhabit isolated localities, complex disjunct distributions, remain poorly understood. Traditionally, vicariance and dispersion have been proposed as explanatory mechanisms that drive such distributions. However, to date, our understanding of the ecological processes driving evolution of ecological niches associated with disjunct distributions remains rudimentary. Here, we propose a framework to deconstruct drivers of such distribution using World's most widespread freshwater fish Galaxias maculatus as a model and integrating marine and freshwater environments where its life cycle may occur. Specifically, we assessed ecological and historical factors (Gondwanan vicariance, marine dispersion) and potential dispersion (niche-tracking) that explain its distribution in the Southern Hemisphere. Estimated distribution was consistent with previously reported distribution and mainly driven by temperature and topography in freshwater environments and by primary productivity and nitrate in marine environments. Niche dynamics of G. maculatus provided evidence of synergy between vicariance and marine dispersion as explanatory mechanisms of its disjunct distribution, suggesting that its ecological niche was conserved since approximately 30 Ma ago. This integrated assessment of ecological niche in marine and freshwater environments serves as a generic framework that may be applied to understand processes underpinning complex distributions of diadromous species.

Los procesos ecológicos que subyacen a las distribuciones de especies que habitan en localidades aisladas, distribuciones disjuntas complejas, siguen siendo poco conocidos. Tradicionalmente, se han propuesto la dispersión y la vicarianza como mecanismos explicativos de tales distribuciones. Sin embargo, hasta la fecha, nuestra comprensión de los procesos ecológicos que impulsan la evolución de los nichos ecológicos de distribuciones disjuntas sigue siendo rudimentaria. Aquí proponemos un marco para de­construir los factores que impulsan dicha distribución, utilizando como modelo el pez de agua dulce con distribución más extendida del mundo, Galaxias maculatus, e integrando los entornos marinos y dulceacuícolas en los que se desarrolla su ciclo vital. En concreto, evaluamos los factores ecológicos e históricos (vicarianza gondwánica, dispersión marina) que explican su distribución en el hemisferio sur. La distribución estimada coincide con la descrita anteriormente para la especie y está determinada principalmente por la temperatura y la topografía en ambientes dulceacuícolas, y la productividad primaria y el nitrato en ambientes marinos. La dinámica de nicho de G. maculatus aportó pruebas de la sinergia entre vicarianza y dispersión marina como mecanismos explicativos de su distribución disjunta, lo que sugiere que su nicho ecológico se conservó desde hace aproximadamente 30 Ma. Esta evaluación integrada del nicho ecológico en ambientes marinos y dulceacuícolas puede aplicarse para comprender los procesos que subyacen a las distribuciones complejas de especies diádromas.

Assessment of the ecological risk and mobility of arsenic and heavy metals in soils and mine tailings from the Carmina mine site (Asturias, NW Spain).

An evaluation of the pollution, distribution, and mobility of arsenic and heavy metals in spoil heaps and soils surrounding the abandoned Carmina lead-zinc mine (Asturias, northern Spain) was carried out. Fractionation of arsenic was performed by an arsenic-specific sequential extraction method; while, heavy metal fractionations was carried out using the protocol of the Bureau Community of Reference (BCR) (now renamed Standards, Measurements and Testing Programme). Arsenic appeared predominantly associated with amorphous iron oxyhydroxides. Among the heavy metals, lead and zinc showed high availability since significant amounts were extracted in the nonresidual fractions; whereas, chromium, copper and nickel showed very low availability, indicating their lithogenic origins. The results showed that the extractability of heavy metals in soils is influenced mainly by the presence of iron and manganese oxides as well as by pH and Eh. Multiple pollution indices, including the enrichment factor (EF), geoaccumulation index (Igeo), ecological risk index (Er) and potential ecological risk index (PERI), were used to assess the degree of soil pollution in the mine area. All results showed that lead was the key factor causing the pollution and ecological risk in the studied area, and copper, zinc and arsenic also had significant contributions. Notably, the sites at higher risk coincided with those with high availability of arsenic and heavy metals. This study provides an integrative approach that serves as a powerful tool to evaluate the metal pollution status and potential threats to the local environment of abandoned mining areas, and the results are useful for making management decisions in these areas.

Ana1 helps recruit Polo to centrioles to promote mitotic PCM assembly and centriole elongation.

Polo kinase (PLK1 in mammals) is a master cell cycle regulator that is recruited to various subcellular structures, often by its polo-box domain (PBD), which binds to phosphorylated S-pS/pT motifs. Polo/PLK1 kinases have multiple functions at centrioles and centrosomes, and we have previously shown that in Drosophila phosphorylated Sas-4 initiates Polo recruitment to newly formed centrioles, while phosphorylated Spd-2 recruits Polo to the pericentriolar material (PCM) that assembles around mother centrioles in mitosis. Here, we show that Ana1 (Cep295 in humans) also helps to recruit Polo to mother centrioles in Drosophila. If Ana1-dependent Polo recruitment is impaired, mother centrioles can still duplicate, disengage from their daughters and form functional cilia, but they can no longer efficiently assemble mitotic PCM or elongate during G2. We conclude that Ana1 helps recruit Polo to mother centrioles to specifically promote mitotic centrosome assembly and centriole elongation in G2, but not centriole duplication, centriole disengagement or cilia assembly. This article has an associated First Person interview with the first author of the paper.

Hyperoxemia in postsurgical sepsis/septic shock patients is associated with reduced mortality.

BACKGROUND: Despite growing interest in treatment strategies that limit oxygen exposure in ICU patients, no studies have compared conservative oxygen with standard oxygen in postsurgical patients with sepsis/septic shock, although there are indications that it may improve outcomes. It has been proven that high partial pressure of oxygen in arterial blood (PaO2) reduces the rate of surgical-wound infections and mortality in patients under major surgery. The aim of this study is to examine whether PaO2 is associated with risk of death in adult patients with sepsis/septic shock after major surgery. METHODS: We performed a secondary analysis of a prospective observational study in 454 patients who underwent major surgery admitted into a single ICU. Patients were stratified in two groups whether they had hyperoxemia, defined as PaO2 > 100 mmHg (n = 216), or PaO2 ≤ 100 mmHg (n = 238) at the day of sepsis/septic shock onset according to SEPSIS-3 criteria maintained during 48 h. Primary end-point was 90-day mortality after diagnosis of sepsis. Secondary endpoints were ICU length of stay and time to extubation. RESULTS: In patients with PaO2 ≤ 100 mmHg, we found prolonged mechanical ventilation (2 [8] vs. 1 [4] days, p < 0.001), higher ICU stay (8 [13] vs. 5 [9] days, p < 0.001), higher organ dysfunction as assessed by SOFA score (9 [3] vs. 7 [5], p < 0.001), higher prevalence of septic shock (200/238, 84.0% vs 145/216) 67.1%, p < 0.001), and higher 90-day mortality (37.0% [88] vs. 25.5% [55], p = 0.008). Hyperoxemia was associated with higher probability of 90-day survival in a multivariate analysis (OR 0.61, 95%CI: 0.39-0.95, p = 0.029), independent of age, chronic renal failure, procalcitonin levels, and APACHE II score > 19. These findings were confirmed when patients with severe hypoxemia at the time of study inclusion were excluded. CONCLUSIONS: Oxygenation with a PaO2 above 100 mmHg was independently associated with lower 90-day mortality, shorter ICU stay and intubation time in critically ill postsurgical sepsis/septic shock patients. Our findings open a new venue for designing clinical trials to evaluate the boundaries of PaO2 in postsurgical patients with severe infections.

Determination of heavy metal baseline levels and threshold values on marine sediments in the Bay of Biscay.

Several international institutions have defined background or baseline levels to assess heavy metal concentrations on marine sediments in order to use these values as a reference for sediment quality indices. This criterion for marine sediment quality is applied to evaluate the potential risk of pollutants in aquatic ecosystems. However, those values were established using samples collected in large areas which present specific geochemical conditions. Then there may be a lack of accuracy in the results when using these parameters in other areas. In this context, 15 sediment cores (8 cm diameter; 2 m length) were recovered along the 400 km Asturian coastline, which is an area with representative lithological conditions for the Bay of Biscay, to determine more precise baseline levels for marine sediments from the Bay of Biscay. An evaluation of statistical and empirical methods was done to determine which method delivers the best results. Statistical methods such as mean±2SD and median±2* Median Absolut Deviation (MAD) are strongly influenced by outliers and data distributions which make these approaches less robust. Graphic techniques such as Cumulative Distribution Function (CDF) avoid the problems that asymmetrical data distributions may cause but introduce a certain level of subjectivity in the results due to the baseline values obtained depending on the researcher's experience. Finally, the Probability Curve (CP) method solves issues which may occur when using other techniques and allows one to establish baseline levels based on different percentiles. Regarding the features of the data analysed in this study, the baselines obtained via the CP method with the 95th percentile appear to be the most accurate for the Bay of Biscay. A wide variation has been found between the new baseline values and other international and national levels. Disparity between those levels and the baselines obtained in this study can be generated by granulometric and geological factors. The notable increase in Hg baseline values with respect to OSPAR Background Concentration values (BCs) (0.05 µg g-1 and 0.6 µg g-1 respectively) and the huge different with CEDEX levels and new threshold levels (0.35 µg g-1 and 1.2 µg g-1 respectively) emphasised the relevance of defining specific baselines and threshold levels, as the ones obtained in this study, not only to obtain more precise criteria for marine sediment quality to be used in environmental assessments, but also to propose new threshold levels for the evaluation of dredged material before dumping into ocean sites.

Interneuron NMDA Receptor Ablation Induces Hippocampus-Prefrontal Cortex Functional Hypoconnectivity after Adolescence in a Mouse Model of Schizophrenia.

Although the etiology of schizophrenia is still unknown, it is accepted to be a neurodevelopmental disorder that results from the interaction of genetic vulnerabilities and environmental insults. Although schizophrenia's pathophysiology is still unclear, postmortem studies point toward a dysfunction of cortical interneurons as a central element. It has been suggested that alterations in parvalbumin-positive interneurons in schizophrenia are the consequence of a deficient signaling through NMDARs. Animal studies demonstrated that early postnatal ablation of the NMDAR in corticolimbic interneurons induces neurobiochemical, physiological, behavioral, and epidemiological phenotypes related to schizophrenia. Notably, the behavioral abnormalities emerge only after animals complete their maturation during adolescence and are absent if the NMDAR is deleted during adulthood. This suggests that interneuron dysfunction must interact with development to impact on behavior. Here, we assess in vivo how an early NMDAR ablation in corticolimbic interneurons impacts on mPFC and ventral hippocampus functional connectivity before and after adolescence. In juvenile male mice, NMDAR ablation results in several pathophysiological traits, including increased cortical activity and decreased entrainment to local gamma and distal hippocampal theta rhythms. In addition, adult male KO mice showed reduced ventral hippocampus-mPFC-evoked potentials and an augmented low-frequency stimulation LTD of the pathway, suggesting that there is a functional disconnection between both structures in adult KO mice. Our results demonstrate that early genetic abnormalities in interneurons can interact with postnatal development during adolescence, triggering pathophysiological mechanisms related to schizophrenia that exceed those caused by NMDAR interneuron hypofunction alone.SIGNIFICANCE STATEMENT NMDAR hypofunction in cortical interneurons has been linked to schizophrenia pathophysiology. How a dysfunction of GABAergic cortical interneurons interacts with maturation during adolescence has not been clarified yet. Here, we demonstrate in vivo that early postnatal ablation of the NMDAR in corticolimbic interneurons results in an overactive but desynchronized PFC before adolescence. Final postnatal maturation during this stage outspreads the impact of the genetic manipulation toward a functional disconnection of the ventral hippocampal-prefrontal pathway, probably as a consequence of an exacerbated propensity toward hippocampal-evoked depotentiation plasticity. Our results demonstrate a complex interaction between genetic and developmental factors affecting cortical interneurons and PFC function.

Seed characterization and early nitrogen metabolism performance of seedlings from Altiplano and coastal ecotypes of Quinoa.

BACKGROUND: Early seed germination and a functional root system development during establishment are crucial attributes contributing to nutrient competence under marginal nutrient soil conditions. Chenopodium quinoa Willd (Chenopodiaceae) is a rustic crop, able to grow in marginal areas. Altiplano and Coastal/Lowlands are two representative zones of quinoa cultivation in South America with contrasting soil fertility and edaphoclimatic conditions. In the present work, we hypothesize that the ecotypes of Quinoa from Altiplano (landrace Socaire) and from Coastal/Lowland (landrace Faro) have developed differential adaptive responses in order to survive under conditions of low availability of N in their respective climatic zones of Altiplano and Lowlands. In order to understand intrinsic differences for N competence between landraces, seed metabolite profile and germinative capacity were studied. Additionally, in order to elucidate the mechanisms of N uptake and assimilation at limiting N conditions during establishment, germinated seeds of both landraces were grown at either sufficient nitrate (HN) or low nitrate (LN) supply. We studied the photosynthetic performance, protein storage, root morphometrical parameters, activity and expression of N-assimilating enzymes, and the expression of nitrate transporters of roots in plants submitted to the different treatments. RESULTS: Seeds from Socaire landrace presented higher content of free N-related metabolites and faster seed germination rate compared to Faro landrace. Seedlings of both ecotypes presented similar physiological performance at HN supply, but at LN supply their differences were exalted. At LN, Socaire plants showed an increased root biomass (including a higher number and total length of lateral roots), a differential regulation of a nitrate transporter (a NPF6.3-like homologue) belonging to the Low Affinity Transport System (LATS), and an upregulation of a nitrate transporter (a NRT2.1-like homologue) belonging to the High Affinity nitrate Transport System (HATS) compared to Faro. These responses as a whole could be linked to a higher amount of stored proteins in leaves, associated to an enhanced photochemical performance in Altiplano plants, in comparison to Lowland quinoa plants. CONCLUSIONS: These differential characteristics of Socaire over Faro plants could involve an adaptation to enhanced nitrate uptake under the brutal unfavorable climate conditions of Altiplano.

Deletion and Randomization of Structurally Variable Regions in B. subtilis Lipase A (BSLA) Alter Its Stability and Hydrolytic Performance Against Long Chain Fatty Acid Esters.

The continuous search for novel enzyme backbones and the engineering of already well studied enzymes for biotechnological applications has become an increasing challenge, especially by the increasing potential diversity space provided by directed enzyme evolution approaches and the demands of experimental data generated by rational design of enzymes. In this work, we propose a semi-rational mutational strategy focused on introducing diversity in structurally variable regions in enzymes. The identified sequences are subjected to a progressive deletion of two amino acids and the joining residues are subjected to saturation mutagenesis using NNK degenerate codons. This strategy offers a novel library diversity approach while simultaneously decreasing enzyme size in the variable regions. In this way, we intend to identify and reduce variable regions found in enzymes, probably resulting from neutral drift evolution, and simultaneously studying the functional effect of said regions. This strategy was applied to Bacillus. subtilis lipase A (BSLA), by selecting and deleting six variable enzyme regions (named regions 1 to 6) by the deletion of two amino acids and additionally randomizing the joining amino acid residues. After screening, no active variants were found in libraries 1% and 4%, 15% active variants were found in libraries 2% and 3%, and 25% for libraries 5 and 6 (n = 3000 per library, activity detected using tributyrin agar plates). Active variants were assessed for activity in microtiter plate assay (pNP-butyrate), thermal stability, substrate preference (pNP-butyrate, -palmitate), and compared to wildtype BSLA. From these analyses, variant P5F3 (F41L-ΔW42-ΔD43-K44P), from library 3 was identified, showing increased activity towards longer chain p-nitrophenyl fatty acid esters, when compared to BSLA. This study allowed to propose the targeted region 3 (positions 40-46) as a potential modulator for substrate specificity (fatty acid chain length) in BSLA, which can be further studied to increase its substrate spectrum and selectivity. Additionally, this variant showed a decreased thermal resistance but interestingly, higher isopropanol and Triton X-100 resistance. This deletion-randomization strategy could help to expand and explore sequence diversity, even in already well studied and characterized enzyme backbones such as BSLA. In addition, this strategy can contribute to investigate and identify important non-conserved regions in classic and novel enzymes, as well as generating novel biocatalysts with increased performance in specific processes, such as enzyme immobilization.

Cerebral Oxygen Saturation and Negative Postoperative Behavioral Changes in Pediatric Surgery: A Prospective Observational Study.

OBJECTIVE: To evaluate if an intraoperative cerebral regional oxygen saturation (crSO2) decrease, less pronounced than 20% below baseline (the current threshold believed to be associated with cognitive dysfunction in adults), is associated with negative postoperative behavioral changes (NPOBC) in the pediatric population after noncardiac surgeries. STUDY DESIGN: A prospective observational study was conducted with 198 children aged 2-12 years old scheduled for noncardiac procedures under general anesthesia. Intraoperatively, crSO2 was monitored with a cerebral oximeter. On postoperative day 7, the Post-Hospital Behavior Questionnaire was used to diagnose NPOBC. RESULTS: The incidence of NPOBC was 38.8%. Logistic regression analysis revealed that with every 1% reduction of crSO2 from the baseline value, the odds of developing NPOBC were 1.199 higher. Likewise, preoperative anxiety (OR 2.832, P = .006), duration of surgery (OR 1.026, P < .0001), and being between the ages of 2 and 3 years (OR 2.604, P = .048) were associated with NPOBC incidence. The multivariable logistic regression model receiver operating characteristic curve showed an area under the curve (95% CI) = 0.820 (0.759-0.881). CONCLUSIONS: During noncardiac surgeries in the pediatric population, an intraoperative decrease in crSO2 less pronounced than 20% from the baseline value is associated with negative postoperative behavior changes on postoperative day 7. The long-term implications remain to be determined, but this supports attention to crSO2 during noncardiac surgeries.