High resolution ultrasound-guided microinjection for interventional studies of early embryonic and placental development in vivo in mice

BACKGROUND: In utero microinjection has proven valuable for exploring the developmental consequences of altering gene expression, and for studying cell lineage or migration during the latter half of embryonic mouse development (from embryonic day 9.5 of gestation (E9.5)). In the current study, we use ultrasound guidance to accurately target microinjections in the conceptus at E6.5–E7.5, which is prior to cardiovascular or placental dependence. This method may be useful for determining the developmental effects of targeted genetic or cellular interventions at critical stages of placentation, gastrulation, axis formation, and neural tube closure. RESULTS: In 40 MHz ultrasound images at E6.5, the ectoplacental cone region and proamniotic cavity could be visualized. The ectoplacental cone region was successfully targeted with 13.8 nL of a fluorescent bead suspension with few or no beads off-target in 51% of concepti microinjected at E6.5 (28/55 injected). Seventy eight percent of the embryos survived 2 to 12 days post injection (93/119), 73% (41/56) survived to term of which 68% (38/56) survived and appeared normal one week after birth. At E7.5, the amniotic and exocoelomic cavities, and ectoplacental cone region were discernable. Our success at targeting with few or no beads off-target was 90% (36/40) for the ectoplacental cone region and 81% (35/43) for the exocoelomic cavity but tended to be less, 68% (34/50), for the smaller amniotic cavity. At E11.5, beads microinjected at E7.5 into the ectoplacental cone region were found in the placental spongiotrophoblast layer, those injected into the exocoelomic cavity were found on the surface or within the placental labyrinth, and those injected into the amniotic cavity were found on the surface or within the embryo. Following microinjection at E7.5, survival one week after birth was 60% (26/43) when the amniotic cavity was the target and 66% (19/29) when the target was the ectoplacental cone region. The survival rate was similar in sham experiments, 54% (33/61), for which procedures were identical but no microinjection was performed, suggesting that surgery and manipulation of the uterus were the main causes of embryonic death. CONCLUSION: Ultrasound-guided microinjection into the ectoplacental cone region at E6.5 or E7.5 and the amniotic cavity at E7.5 was achieved with a 7 day postnatal survival of ≥60%. Target accuracy of these sites and of the exocoelomic cavity at E7.5 was ≥51%. We suggest that this approach may be useful for exploring gene function during early placental and embryonic development

The 2009 multiwavelength campaign on Mrk 421: Variability and correlation studies

We performed a 4.5-month multi-instrument campaign (from radio to VHE gamma rays) on Mrk421 between January 2009 and June 2009, which included VLBA, F-GAMMA, GASP-WEBT, Swift, RXTE, Fermi-LAT, MAGIC, and Whipple, among other instruments and collaborations. Mrk421 was found in its typical (non-flaring) activity state, with a VHE flux of about half that of the Crab Nebula, yet the light curves show significant variability at all wavelengths, the highest variability being in the X-rays. We determined the power spectral densities (PSD) at most wavelengths and found that all PSDs can be described by power-laws without a break, and with indices consistent with pink/red-noise behavior. We observed a harder-when-brighter behavior in the X-ray spectra and measured a positive correlation between VHE and X-ray fluxes with zero time lag. Such characteristics have been reported many times during flaring activity, but here they are reported for the first time in the non-flaring state. We also observed an overall anti-correlation between optical/UV and X-rays extending over the duration of the campaign. The harder-when-brighter behavior in the X-ray spectra and the measured positive X-ray/VHE correlation during the 2009 multi-wavelength campaign suggests that the physical processes dominating the emission during non-flaring states have similarities with those occurring during flaring activity. In particular, this observation supports leptonic scenarios as being responsible for the emission of Mrk421 during non-flaring activity. Such a temporally extended X-ray/VHE correlation is not driven by any single flaring event, and hence is difficult to explain within the standard hadronic scenarios. The highest variability is observed in the X-ray band, which, within the one-zone synchrotron self-Compton scenario, indicates that the electron energy distribution is most variable at the highest energies.Comment: Accepted for publication in A&A, 18 pages, 14 figures (v2 has a small modification in the acknowledgments, and also corrects a typo in the field "author" in the metadata

miR-27b inhibits fibroblast activation via targeting TGFB signaling pathway

Background: MicroRNAs are a group of small RNAs that regulate gene expression at the posttranscriptional level. They regulate almost every aspect of cellular processes. In this study, we investigated whether miR-27b regulates pulmonary fibroblast activation.Results: We found that miR-27b was down-regulated in fibrotic lungs and fibroblasts from an experimental mouse model of pulmonary fibrosis. The overexpression of miR-27b with a lentiviral vector inhibited TGFB1-stimulated mRNA expression of collagens (COL1A1, COL3A1, and COL4A1) and alpha-smooth muscle actin, and protein expression of Col3A1 and alpha-smooth muscle actin in LL29 human pulmonary fibroblasts. miR-27b also reduced contractile activity of LL29. TGFB receptor 1 and SMAD2 were identified as the targets of miR-27b by 3'-untranslated region luciferase reporter and western blotting assays.Conclusions: Our results suggest that miR-27b is an anti-fibrotic microRNA that inhibits fibroblast activation by targeting TGFB receptor 1 and SMAD2. This discovery may provide new targets for therapeutic interventions of idiopathic pulmonary fibrosis.Peer reviewedPhysiological SciencesOklahoma Center for Respiratory and Infectious Disease

Recent progress in low-carbon binders

The development of low-carbon binders has been recognized as a means of reducing the carbon footprint of the Portland cement industry, in response to growing global concerns over CO2 emissions from the construction sector. This paper reviews recent progress in the three most attractive low-carbon binders: alkali-activated, carbonate, and belite-ye'elimite-based binders. Alkali-activated binders/materials were reviewed at the past two ICCC congresses, so this paper focuses on some key developments of alkali-activated binders/materials since the last keynote paper was published in 2015. Recent progress on carbonate and belite-ye'elimite-based binders are also reviewed and discussed, as they are attracting more and more attention as essential alternative low-carbon cementitious materials. These classes of binders have a clear role to play in providing a sustainable future for global construction, as part of the available toolkit of cements

Organisch-anorganische Wechselwirkungen in siliziklastischen Ölreservoiren in der Nordsee

Sandstones are important reservoir rocks for a variety of fluids. Their properties, such as porosity and permeability, which are crucial factors for petroleum production, primarily depend on depositional environments. Additionally, subsequent geochemical alterations by depth-related reactions (“classic diagenesis”), organic-inorganic interactions, and mixing of basin fluids can also significantly alter reservoir properties via mineral dissolution and (re-)precipitation. However, the impact of organic-inorganic interactions on mineral alteration is still poorly understood and the controlling factors for meteoric water intrusions are unknown. It is the aim of this dissertation to investigate the diagenesis of sandstones in order to figure out how organic-inorganic interactions affected mineral assemblages depending on oil degradation of different intensity, on mineralogical composition and on pore water composition. Furthermore, the controlling factors for meteoric waters intruding into deep marine turbidites and their consequence on organic-inorganic interactions were studied. To achieve these objectives, more than one hundred core samples were taken from four oil fields (Siri field, Magnus field, Frigg field and Grane field) in the North Sea. These samples vary in lithology (glauconite sandstone, arkose sandstone, and subarkose sandstone), and cover an age range of Jurassic, Palaeocene and Eocene, and a depth interval from shallow (1,734-1,798 m) to deep (3,200-3,240 m). An interdisciplinary approach including imaging techniques (thin section, SEM, TEM), inorganic geochemical techniques (XRD and XRF) and organic geochemical methodologies (GC-FID, GC-MS, FT-ICR-MS) was used for analyzing the inorganic and organic constituents of the samples, and their genetic relationships. In the glauconite-bearing Heimdal sandstone reservoir of the Siri field (2,096-2,137 m depth), authigenic berthierine is one of the most significant diagenetic phases. Berthierine mainly appears as pore-lining or pore-filling crystals, and is most abundant at the oil-water contact (OWC) where glauconite is relatively depleted and where extracted bitumens show the highest extent of biodegradation. Berthierine commonly occurs in semi-dissolved glauconite and its formation postdates oil filling. Based on these two observations, it can be inferred that the formation of berthierine was result of glauconite dissolution, and that this process was probably driven by organic-inorganic interactions due to oil degradation at the OWC. Due to these interconnected processes, other diagenetic alterations, such as feldspar dissolution and quartz overgrowth, also take place in the Heimdal glauconite-bearing sandstone. In the Magnus field, the Jurassic Magnus sandstone is the main reservoir which is at present deeply buried at depths of ca. 3,200-3,240 m. The Magnus sandstone is an arkose sandstone with a high feldspar content. This sandstone reservoir has experienced various intensive post-depositional alterations, including quartz overgrowth, formation of ankerite, siderite, albite, and kaolinite, as well as K-feldspar dissolution. Petrographic and geochemical analyses indicate that part of albite was formed within the reservoir at temperatures of around 80-100°C after oil emplacement. Three potential mechanisms for albite formation were proposed and tested by hydrogeochemical modelling approaches. The results indicate that (1) dissolution of unstable minerals (such as kaolinite and chalcedony) coupled to reduction of ferric iron minerals, but also (2) dissolution of non-end member feldspar coupled to illite formation can account for albite formation. The Eocene Frigg sandstones of the Frigg field and the Palaeocene Heimdal sandstones of the Grane field are both sub-arkose sandstones and are buried at 2,045-2,075 m and 1,734-1,798 m, respectively. However, the Eocene Frigg sandstones display intensive feldspar dissolution and kaolinite formation, and stronger oil degradation compared to the Palaeocene Heimdal sandstones of the Grane field. Most likely, meteoric water intrusion into the Frigg sandstones due to its adjacency of the East Shetland Platform could have led to both alterations. Furthermore, organic-inorganic interactions triggered by oil degradation could also be a reason for feldspar dissolution and kaolinite formation accompanied by siderite formation.Sandstein ist ein wichtigstes Reservoirgestein für Fluide. Seine Eigenschaften, wie zum Beispiel Porosität und Permeabilität, die entscheidend für die Erdöl- und Erdgasproduktion sind, hängen stark vom Ablagerungsmilieu ab. Zusätzlich können nachfolgende geochemische Prozesse wie z.B. tiefenabhängige Reaktionen (“Klassische Diagenese”), organisch-anorganische Wechselwirkungen, und Vermischungen von unterschiedlichen Beckenfluiden die Reservoir-Eigenschaften durch Lösung und Ausfällung von Mineralen erheblich verändern. Trotzdem ist der Einfluss organisch-anorganischer Wechselwirkungen auf Mineralveränderungen immer noch wenig erforscht und die kontrollierenden Faktoren von Intrusionen meteorischen Wassers weitestgehend unbekannt. Das Ziel der vorliegenden Dissertation ist die Untersuchung der Sandsteindiagenese in Bezug auf den Einfluss organisch-anorganischer Wechselwirkungen auf Mineralvergesellschaftungen und deren Abhängigkeit von der Intensität des Erdölabbaus, der primären mineralogischen Zusammensetzung, und von der Porenwasser-Zusammensetzung. Weiterhin wurden die Einflussfaktoren von Intrusionen meteorischen Wassers in tiefmarine Turbidite und deren Folgen auf organisch-anorganischen Wechselwirkungen untersucht. Hierzu wurden mehr als 100 Kernproben aus vier Ölfeldern in der Nordsee genommen (Siri Feld, Magnus Feld, Frigg Feld und Grane Feld). Diese Proben variieren im Hinblick auf Lithologie (glaukonitscher Sandstein, Arkose-Sandstein, subarkosischer Sandstein), Alter (von Jura, Paläozän, bis Eozän), und Tiefe, wo sie sich von flachen (1,734-1,798 m) bis zu tiefen (3,200-3,240 m) Bereichen erstrecken. Um organische und anorganische Bestandteile der Proben und deren genetische Zusammenhänge zu analysieren, wurde ein interdisziplinärer Ansatz mit bildgegebenden Verfahren (Dünnschliff, SEM, TEM), anorganisch-geochemischen (XRD und XRF), und organisch-geochemischen Methoden (GC-FID, GC-MS, FT-ICR-MS) angewendet. Im Glaukonit-haltigen Heimdal-Sandstein des Siri-Reservoirs (2,096-2,137 m) ist authigener Berthierin eine der bedeutendsten diagenetischen Mineralphasen. Berthierin kommt hauptsächlich als Porensaum oder Porenfüllung vor und tritt zumeist am Öl-Wasser-Kontakt (OWC) auf. Hier ist Glaukonit relativ verarmt und extrahierte Bitumina zeigen das höchste Biodegradationsstadium. Berthierin tritt normalerweise in angelöstem Glaukonit auf und seine Bildung eröffnet die Möglichkeit, die Füllungsgeschichte des Reservoirs zu datieren. Aus beiden Beobachtungen lässt sich schließen, dass der Übergang von Glaukonit zu Berthierin durch organisch-inorganische Wechselwirkungen verursacht wird und im Umkehrschluss durch die Öldegradation am OWC angetrieben wird. Aufgrund dieser miteinander verbundenen Prozesse finden zudem andere diagenetische Prozesse in den Glaukonit-haltigen Heimdal-Sandsteinen statt, zum Beispiel Auflösung von Feldspäten und Quarz-Überwachsung. Jurassische Magnus Sandsteine sind das Hauptreservoir im Magnus-Feld und bis in 3,200-3,240 m Tiefe versenkt. Diese bestehen aus Arkosesandsteinen mit hohem Feldspat-Anteil. Das Reservoir hat verschiedene, intensive Prozesse nach der Ablagerung erlebt, wie Quarz-Überwachsung, Neubildung von Ankerit, Siderit, Albit, sowie Auflösung vom Kalifeldspat. Petrographische und geochemische Untersuchungen weisen darauf hin, dass ein Teil des Albits bei Temperaturen von ca. 80-100°C im Reservoir gebildet wurde, nachdem das Öl eingedrungen ist. Drei mögliche Mechanismen für die Albit-Bildung wurden mit Hilfe von hydrogeochemischen Modellierungen überprüft. Die modellierten Ergebnisse deuten darauf hin, dass (1) die Auflösung von instabilen Mineralen (wie Kaolinit und Chalzedonit) mit der Reduktion Eisen(III)-haltiger Minerale verbunden sind, und (2) dass die Neubildung vom Albit auch durch Auflösung von Feldspatmischkristallen und gleichzeitiger Illit-Bildung verantwortlich sein kann. Die eozänen Sandsteine des Frigg Feldes und die paläozänen Sandsteine des Grane Felds sind beide Subarkosesandsteine und treten in Tiefen von jeweils 2,045-2,075 m und 1,734-1,798 m auf. Jedoch zeigen die Frigg Sandsteine eine intensivere Feldspatauflösung, eine stärkere Kaolinitbildung, sowie eine stärkere Erdöldegradation im Vergleich zu den paläozänen Heimdal-Sandsteinen des Grane Feldes. Es wird vermutet, dass die Intrusion meteorischen Wassers in die Frigg-Sandsteine aufgrund seiner Nähe zur East Shetland Plattform stattfand, und dies zu beiden genannten Alterationsprozessen geführt hat. Weiterhin könnten die vom Ölabbau ausgelösten organisch-anorganischen Interaktionen eine Ursache für Feldspatauflösungen und Bildung von Kaolinit sowie Siderit sein. Schlüsselwörter: Organisch-anorganische Wechselwirkung, Sandstein, Glaukonit, Berthierin, Albit, meteorische Wässer, FT-ICR-MS, Nordsee

Stereoselective Separation and Acute Toxicity of Tau-Fluvalinate to Zebrafish

Tau-fluvalinate (TFLV) is one of the most potent chiral synthetic pyrethroids to control a wide range of pests in agricultural fields, especially in apiary. In this study, two stereoisomers of TFLV were fully separated by high-performance liquid chromatography (HPLC) with a semipreparative chiral column using cellulose-tris(3,5-dimethylphenylcarbamate) as chiral stationary phase and n-hexane and 2-propanol (96/4, v/v) as mobile phase at a flow rate of 2.5 mL min−1. The (+)-stereoisomer was first eluted by detecting with an optical rotation detector. After obtaining pure single stereoisomer of TFLV, acute toxicities of each isomer and TFLV standard to zebrafish were evaluated. The results showed that the (+)-stereoisomer exhibited 273.4 times higher toxicity than the (−)-stereoisomer and 6.7 times higher than TFLV standard, according to their LC50 values at 96 h after exposure. This indicates that the toxicity of TFLV mainly originates from (+)-stereoisomer. These results are very helpful to prepare single stereoisomer of chiral pesticides and evaluate their different toxicological effects to aquatic organisms

Investigation of Rainfall-Runoff Processes and Soil Moisture Dynamics in Grassland Plots under Simulated Rainfall Conditions

The characteristics of rainfall-runoff are important aspects of hydrological processes. In this study, rainfall-runoff processes and soil moisture dynamics at different soil depths and slope positions of grassland with two different row spacings (5 cm and 10 cm, respectively, referred to as R5 and R10) were analyzed, by means of a solution of rainfall simulation experiments. Bare land was also considered as a comparison. The results showed that the mechanism of runoff generation was mainly excess infiltration overland flow. The surface runoff amount of R5 plot was greater than that of R10, while the interflow amount of R10 was larger than that of R5 plot, although the differences of the subsurface runoff processes between plots R5 and R10 were little. The effects of rainfall intensity on the surface runoff were significant, but not obvious on the interflow and recession curve, which can be described as a simple exponential equation, with a fitting degree of up to 0.854–0.996. The response of soil moisture to rainfall and evapotranspiration was mainly in the 0–20 cm layer, and the response at the 40 cm layer to rainfall was slower and generally occurred after the rainfall stopped. The upper slope generally responded fastest to rainfall, and the foot of the slope was the slowest. The results presented here could provide insights into understanding the surface and subsurface runoff processes and soil moisture dynamics for grasslands in semi-arid regions

Pore Structure Characteristics of Shale Oil Reservoirs with Different Lithofacies and Their Effects on Mobility of Movable Fluids: A Case Study of the Chang 7 Member in the Ordos Basin, China

The Chang 7 member of the Triassic Yanchang Formation in the Ordos Basin is a significant continent shale oil reservoir in China. Therefore, conducting an in-depth investigation into the pore structure and fluid mobility characteristics of the Chang 7 shale oil reservoir holds immense importance for advancing shale oil exploration. This study conducts a detailed analysis of the pore structures and their impact on fluid mobility of the Chang 7 shale oil reservoir using multiple methodologies, i.e., a cast thin section, scanning electron microscopy (SEM), X-ray diffraction (XRD), high-pressure mercury injection (HPMI), low-temperature nitrogen adsorption (LTNA), and nuclear magnetic resonance (NMR). The results show that the sandstone in the Yanwumao area of the Chang 7 shale oil reservoir consists mainly of lithic arkose and feldspathic litharenite, which can be classified into three lithofacies (massive fine-grained sandstone (Sfm), silt-fine sandstone with graded bedding (Sfgb), and silt-fine sandstone with parallel bedding (Sfp)). Moreover, three pore structures (Type I, II, and III), and four pore spaces (nanopores, micropores, mesopores, and macropores) can be characterized. Pore structure Type I, characterized by large pores, exhibits bimodal pore diameter curves, resulting in the highest levels of movable fluid saturation (MFS) and movable fluid porosity (MFP). Pore structure Type II demonstrates unimodal pore structures, indicating robust connectivity, and higher MFS and MFP. Pore structure Type III primarily consists of dissolved and intercrystalline pores with smaller pore radii, a weaker pore configuration relationship, and the least fluid mobility. Furthermore, a correlation analysis suggests that the pore structure significantly impacts the fluid flowability in the reservoir. Favorable petrophysical properties and large pores enhance fluid flowability. Micropores and mesopores with high fractal dimensions have a greater impact on reservoir fluid mobility compared to macropores and nanopores. Mesopores mainly control MFS and MFP, while micropores govern the shift from bound fluid to movable fluid states. Among the lithofacies types, the Sfm lithofacies exhibit the highest fluid mobility due to their significant proportion of macropores and mesopores, whereas the Sfgb lithofacies have lower values because they contain an abundance of micropores. The Sfp lithofacies also dominate macropores and mesopores, resulting in medium fluid mobility levels. This study combines lithofacies types, micro-reservoir pore structure characteristics, and mobile fluid occurrence characteristics to better understand the dominant reservoir distribution characteristics of the Chang 7 shale oil reservoirs in the Ordos Basin and provide theoretical information for further optimization of production strategies

Effects of Rainfall Intensity and Slope Gradient on Runoff and Soil Moisture Content on Different Growing Stages of Spring Maize

The rainfall-runoff process (RRP) is an important part of hydrologic process. There is an effective measure to study RRP through artificial rainfall simulation. This paper describes a study on three growing stages (jointing stage, tasseling stage, and mature stage) of spring maize in which simulated rainfall events were used to study the effects of various factors (rainfall intensity and slope gradient) on the RRP. The RRP was tested with three different rainfall intensities (0.67, 1.00, and 1.67 mm/min) and subjected to three different slopes (5°, 15°, and 20°) so as to study RRP characteristics in semiarid regions. Regression analysis was used to study the results of this test. The following key results were obtained: (1) With the increase in rainfall intensity and slope, the increasing relationship with rainfall duration, overland flow, and cumulative runoff, respectively, complied with logarithmic and quadratic functions before reaching stable runoff in each growing stage of spring maize; (2) The runoff coefficient increased with the increase in rainfall intensity and slope in each growing stages of spring maize. The relationship between runoff coefficient, slope, rainfall intensity, rainfall duration, antecedent soil moisture, and vegetation coverage was multivariate and nonlinear; (3) The runoff lag time decreased with the increase in rainfall intensity and slope within the same growing stage. In addition, the relationship between runoff lag time, slope, rainfall intensity, antecedent soil moisture, and vegetation coverage could also be expressed by a multivariate nonlinear equation; (4) The descent rate of soil infiltration rate curve increased with the increased rainfall intensity and slope in the same growing stage. Furthermore, by comparing the Kostiakov, Horton, and Philip models, it was found that the Horton infiltration model was the best for estimating soil infiltration rate and cumulative infiltration under the condition of test

Analysis of the Enantioselective Effects of PCB95 in Zebrafish (Danio rerio) Embryos through Targeted Metabolomics by UPLC-MS/MS.

As persistent organic pollutants, polychlorinated biphenyls (PCBs) accumulate in the bodies of animals and humans, resulting in toxic effects on the reproductive, immune, nervous, and endocrine systems. The biological and toxicological characteristics of enantiomers of chiral PCBs may differ, but these enantioselective effects of PCBs have not been fully characterized. In this study, we performed metabolomics analysis, using ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) to investigate the enantioselective toxic effects of PCB95 in zebrafish (Danio rerio) embryos after exposure to three dose levels of 0.1, 1, and 10 μg/L for 72 h. Multivariate analysis directly reflected the metabolic perturbations caused by PCB95. The effects of (-)-PCB95 and (+)-PCB95 were more prominent than those of the racemate in zebrafish embryos. A total of 26 endogenous metabolites were selected as potential marker metabolites with variable importance at projection values larger than 1 and significant differences (p<0.05). These metabolites included amino acids, organic acids, nucleosides, betaine, and choline. The changes in these biomarkers were dependent on the enantiomer-specific structures of PCB95. Fifteen metabolic pathways were significantly affected, and several nervous and immune system-related metabolites were significantly validated after exposure. These metabolic changes indicated that the toxic effects of PCB95 may be associated with the interaction of PCB95 with the nervous and immune systems, thus resulting in disruption of energy metabolism and liver function