Clustered regularly interspaced short palindromic repeats and CRISPR‐associated protein‐9 (CRISPR‐Cas9) can be used as an efficient tool for genome editing in potato (Solanum tuberosum). From both a ...scientific and a regulatory perspective, it is beneficial if integration of DNA in the potato genome is avoided. We have implemented a DNA‐free genome editing method, using delivery of CRISPR‐Cas9 ribonucleoproteins (RNPs) to potato protoplasts, by targeting the gene encoding a granule bound starch synthase (GBSS, EC 2.4.1.242). The RNP method was directly implemented using previously developed protoplast isolation, transfection and regeneration protocols without further adjustments. Cas9 protein was preassembled with RNA produced either synthetically or by in vitro transcription. RNP with synthetically produced RNA (cr‐RNP) induced mutations, i.e. indels, at a frequency of up to 9%, with all mutated lines being transgene‐free. A mutagenesis frequency of 25% of all regenerated shoots was found when using RNP with in vitro transcriptionally produced RNA (IVT–RNP). However, more than 80% of the shoots with confirmed mutations had unintended inserts in the cut site, which was in the same range as when using DNA delivery. The inserts originated both from DNA template remnants from the in vitro transcription, and from chromosomal potato DNA. In 2–3% of the regenerated shoots from the RNP‐experiments, mutations were induced in all four alleles resulting in a complete knockout of the GBSS enzyme function.
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•Aqueous extract of Cystoseira baccata served to synthesize gold nanoparticles Au@CB.•Au@CB are spherical polycrystalline nanoparticles with a mean diameter of 8.4nm.•Au@CB exhibit ...strong cytotoxicity against Caco-2 and HT-29 and no effect on the healthy cells.•Au@CB induce apoptosis activation by the extrinsic and mitochondrial pathway.
This study is the first dealt with the use of brown macroalgae Cystoseira baccata (CB) extracts in obtaining gold nanoparticles (Au@CB) through an eco-friendly, fast, one-pot synthetic route. The formation of spherical, stable, polycrystalline nanoparticles with mean diameter of 8.4±2.2nm was demonstrated by UV–vis spectroscopy, TEM, HRTEM, STEM and zeta potential measurements. The extract appears to act as a protective agent where the particles are embedded, keeping them separated, avoiding aggregation and coalescence. The EELS and EDS analyses confirmed the elemental composition of the extract and nanoparticles. Moreover, the functional group of biomolecules present in CB and Au@CB were characterized by FTIR. The effects of CB extract and Au@CB were tested in vitro on the colon cancer cell lines HT-29 and Caco-2, as well as on normal primary neonatal dermal fibroblast cell line PCS-201-010. Results show a stronger cytotoxic effect against HT-29 than that on Caco-2; interestingly, a lack of toxicity on PCS-201-010 was obtained. Finally, the apoptotic activity was determined; Au@CB is able to induce apoptosis activation by the extrinsic and mitochondrial pathway in our CRC in vitro model. These encouraging results suggest that Au@CB has a significant potential for the treatment of colon rectal cancer.
Abstract Objective To investigate whether pregestational diabetes mellitus (DM) induces changes in vascular placental development detectable at first trimester. Methods This was a prospective ...case–control study in 69 women with pregestational DM and 94 controls undergoing first-trimester combined screening for aneuploidies. Maternal characteristics, fetal nuchal translucency thickness, maternal serum pregnancy-associated plasma protein A (PAPP-A) and free β human chorionic gonadotrophin (β-hCG) were evaluated. Three-dimensional ultrasound was used to measure placental volume and three dimensional power Doppler (3D-PD) placental vascular indices including: vascularization index (VI), flow index (FI) and vascularization flow index (VFI). Pregnancy-associated hypertensive complications (PAHC) and perinatal outcomes were analyzed. The total group of diabetic women and the group of diabetic women without PAHC were compared separately with the control group. Results 3D-PD placental vascular indexes were significantly lower in women with DM than in controls (VI p = 0.007, FI p = 0.003 and VFI p = 0.04). These differences remained on excluding cases with PAHC in the DM group. No differences were found in placental volumes between the DM group and controls. Serum PAPP-A levels were also lower in diabetic women ( p < 0.02) and negatively correlated with the degree of maternal metabolic control at first trimester. Conclusions Pregestational DM induces demonstrable alterations in first trimester placental development, with significantly reduced placental vascularization indices and PAPP-A values. This effect is independent of the later development of PAHC.
In this paper we analyze the evolution of Latin American (LATAM) Business Economics (BE) publications in international journals from 2005 to 2019. Using publications in Web of Science Core Collection ...(WoS), we analyze which characteristics of collaboration result in higher impact, i.e., total number of citations, and journals’ WoS impact factor. Our findings show that the number of publications in journals indexed in the WoS by researchers in LATAM have been rising in terms of the number of publications and impact measured by citations. Moreover, researchers in the region are publishing in journals with higher impact factor. The analysis shows that the main drivers of impact are multilateral and bilateral collaboration, number of countries, number of authors, and the number of categories of knowledge. Specifically, multilateral collaboration is a key factor of influential papers. Other aspects that increase the impact of publications are publishing in English and collaborating with authors from the United States. Our results also suggest a slight decrease in the impact as the number of coauthors increase.
A growing interest in extreme precipitation has spread through the scientific community due to the effects of global climate change on the hydrological cycle, and their threat to natural systems' ...higher than average climatic values. Understanding the variability of precipitation indices and their association to atmospheric processes could help to project the frequency and severity of extremes. This paper evaluates the trend of three precipitation extremes: the number of consecutive dry/wet days (CDD/CWD) and the quotient of the precipitation in days where daily precipitation exceeds the 95th percentile of the reference period and the total amount of precipitation (or contribution of very wet days, R95pTOT). The aim of this study is twofold. First, extreme indicators are compared against accumulated precipitation (RR) over Europe in terms of trends using non-parametric approaches. Second, we analyse the geographically opposite trends found over different parts of Europe by considering their relationships with large-scale processes, using different teleconnection patterns. The study is accomplished for the four seasons using the gridded E-OBS data set developed within the EU ENSEMBLES project. Different patterns of variability were found for CWD and CDD in winter and summer, with north-south and east-west configurations, respectively. We consider physical factors in order to understand the extremes' variability by linking large-scale processes and precipitation extremes. Opposite associations with the North Atlantic Oscillation in winter and summer, and the relationships with the Scandinavian and East Atlantic patterns as well as El Niño/Southern Oscillation events in spring and autumn gave insight into the trend differences. Significant relationships were found between the Atlantic Multidecadal Oscillation and R95pTOT during the whole year. The largest extreme anomalies were analysed by composite maps using atmospheric variables and sea surface temperature. The association of extreme precipitation indices and large-scale variables found in this work could pave the way for new possibilities regarding the projection of extremes in downscaling techniques.
Thyroid hormones participate in regulating growth and homeostatic processes in vertebrates, including development and adult functioning of the reproductive system. Here we report a new stimulatory ...role of thyroid hormone on the proliferation of Sertoli cells (SCs) and single, type A undifferentiated spermatogonia (Aund) in adult zebrafish testes. A role for T3 in zebrafish testis is suggested by in situ hybridization studies, which localized thyroid receptor α (thrα) in SCs and the β (thrβ) mRNA in Sertoli and Leydig cells. Using a primary zebrafish testis tissue culture system, the effect of T3 on steroid release, spermatogenesis, and the expression of selected genes was evaluated. Basal steroid release and Leydig cell gene expression did not change in response to T3. However, in the presence of FSH, T3 potentiated gonadotropin-stimulated androgen release as well as androgen receptor (ar) and 17α-hydroxylase/17,20 lyase (cyp17a1) gene expression. Moreover, T3 alone stimulated the proliferation of both SCs and Aund, potentially resulting in newly formed spermatogonial cysts. Additional tissue culture studies demonstrated that Igf3, a new, gonad-specific member of the IGF family, mediated the stimulatory effect of T3 on the proliferation of Aund and SCs. Finally, T3 induced changes in connexin 43 mRNA levels in the testis, a known T3-responsive gene. Taken together, our studies suggest that T3 expands the population of SCs and Aund involving Igf signaling and potentiates gonadotropin-stimulated testicular androgen production as well as androgen sensitivity.
Pre-operative administration of benzodiazepines can cause hypoventilation-a decrease in minute ventilation (MV)-commonly referred to as "respiratory compromise or respiratory depression." Respiratory ...depression can lead to hypercarbia and / or hypoxemia, and may heighten the risk of other respiratory complications. Current anesthesia practice often places patients at risk for respiratory complications even before surgery, as respiratory monitoring is generally postponed until the patient is in the operating room. In the present study we examined and quantified the onset of respiratory depression following the administration of a single dose of midazolam in pre-operative patients, using a non-invasive respiratory volume monitor that reports MV, tidal volume (TV), and respiratory rate (RR).
Impedance-based Respiratory Volume Monitor (RVM) data were collected and analyzed from 30 patients prior to undergoing orthopedic or general surgical procedures. All patients received 2.0 mg of midazolam intravenously at least 20 minutes prior to the induction of anesthesia and the effects of midazolam on the patient's respiratory function were analyzed.
Within 15 minutes of midazolam administration, we noted a significant decrease in both MV (average decrease of 14.3% ± 5.9%, p<0.05) and TV (22.3% ± 4.5%, p<0.001). Interestingly, the corresponding RR increased significantly by an average of 10.3% ± 4.7% (p<0.05). Further analysis revealed an age-dependent response, in which elderly patients (age≥65 years, n = 6) demonstrated greater reductions in MV and TV and a lack of compensatory RR increase. In fact, elderly patients experienced an average decrease in MV of 34% ± 6% (p<0.05) compared to an average decrease of 9% ± 6% (p<0.05) in younger patients.
We were able to quantify the effects of pre-operative midazolam administration on clinically significant respiratory parameters (MV, TV and RR) using a non-invasive RVM, uncovering that the respiratory depressive effect of benzodiazepines affect primarily TV rather than RR. Such respiratory monitoring data provide the opportunity for individualizing dosing and adjustment of clinical interventions, especially important in elderly patients. With additional respiratory data, clinicians may be able to better identify and quantify respiratory depression, reduce adverse effects, and improve overall patient safety.
•Results suggest that the network dynamics behind the generation of quality research output contrasts dramatically with that of quantity.•We find that the relational dimension of scientists matters ...for quality, but not for output.•The cognitive dimensions have the opposite effect, helping research output, while being indifferent towards impact.•The structural dimension of the network is the only area where there is some degree of convergence between output quantity and quality.
This paper aims to further our understanding of how embeddedness affects the research output and impact of scientists. The analysis uses an extensive panel data that allows an analysis of within person variation over time. It explores the simultaneous effects of different dimensions of network embeddedness over time at individual level. These include the establishment of direct ties, the strengths of these ties, as well as the density, structural holes, centrality, and cross-disciplinary links. Results suggest that the network dynamics behind the generation of quality output contrasts dramatically with that of quantity. We find that the relational dimension of scientists matters for quality, but not for output, while cognitive dimensions have the opposite effect, helping output, while being indifferent toward impact. The structural dimension of the network is the only area where there is some degree of convergence between output quantity and quality; here, we find a prevalence for the role of brokerage over cohesion. It concludes by discussing implications for both network research and science policy.
The Scientific Impact of Developing Nations Gonzalez-Brambila, Claudia N; Reyes-Gonzalez, Leonardo; Veloso, Francisco ...
PloS one,
03/2016, Volume:
11, Issue:
3
Journal Article
Peer reviewed
Open access
This paper analyzes science productivity for nine developing countries. Results show that these nations are reducing their science gap, with R&D investments and scientific impact growing at more than ...double the rate of the developed world. But this "catching up" hides a very uneven picture among these nations, especially on what they are able to generate in terms of impact and output relative to their levels of investment and available resources. Moreover, unlike what one might expect, it is clear that the size of the nations and the relative scale of their R&D investments are not the key drivers of efficiency.
The occurrence, distribution and bioaccumulation of five endocrine disrupting compounds (4-tert-octylphenol, 4-n-octylphenol, 4-n-nonylphenol, nonylphenol and bisphenol A) in water, sediment and ...biota (Corbicula fluminea) collected along the Minho River estuary (NW Iberian Peninsula) were examined. Samples were collected in two campaigns (May and November, 2012) and analyzed by different extraction procedures followed by liquid chromatography tandem mass spectrometry determination. The presence of linear isomers (4-n-octylphenol and 4-n-nonylphenol) was scarcely observed whereas branched isomers (4-tert-octylphenol and nonylphenol) were measured in almost all samples. Wastewater treatment plant effluents and nautical, fishing and agricultural activities are considered the primary source of pollution of the river by alkylphenols. The presence of bisphenol A in the river could be mainly associated to punctual sources of contamination from industrial discharges. A decrease in the total concentration of phenolic compounds in water was observed from spring to autumn (from 0.888μgL−1 in May to 0.05μgL−1 in November), while similar values were shown in C. fluminea samples from the two campaigns (1388 and 1228ngg−1 dw in spring and autumn, respectively). In sediments, the total concentration of the target compounds varied between 13 and 4536ngg−1 dw (average of 1041ngg−1 dw). Sediment–water partition coefficient (Kd), bioaccumulation factor (BAF) and biota–sediment accumulation factor (BSAF) were estimated and highest values were obtained for nonylphenol. Calculated risk quotients showed low and moderate risk for the aquatic environment from the presence of the target compounds at all sampling points. The estimation of the daily intake of the studied compounds via water and biota ingestion indicated no risk for human health.
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•Occurrence of 5 EDCs (APs and BPA) in water, sediment and biota from the Minho River•Higher EDC concentrations in water were observed during the spring campaign.•Low levels of EDCs measured in sediment and biota samples, except in the case of NP•Bioaccumulation factors and sediment–water partition coefficients were estimated.•Ecological and human health impacts were estimated and no risk was identified.
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