Improved methods for detection of β-galactosidase (lacZ) activity in hard tissue

The ß-galactosidase gene (lacZ) of Escherichia coli is widely used as a reporter gene. The expression of lacZ can be detected by enzyme-based histochemical staining using chromogenic substrates such as 5-bromo-4-chloro-3-indolyl-ß-D: -galactoside (X-gal). Because the enzymatic activity of lacZ is vulnerable to high temperatures and acid treatment for demineralization, detection of lacZ on paraffinized sections is difficult, especially for hard tissues, which require demineralization before sectioning in paraffin. To circumvent this problem, whole-mount X-gal staining before sectioning is performed. However, detection of lacZ activity in the center of larger portions of hard whole adult tissues is challenging. In this study, focusing on fixation procedures, we determined the conditions conducive to improved detection of lacZ activity in deeper areas of whole tissues. We used an annexin a5 (Anxa5)-lacZ reporter mouse model in which the Anxa5 expression in hard tissue is indicated by lacZ activity. We found that lacZ activity could be detected throughout the periodontal ligament of adult mice when fixed in 100% acetone, whereas it was not detected in the periodontal ligament around the root apex fixed in glutaraldehyde and paraformaldehyde. This staining could not be detected in wild-type mice. Acetone maintains the lacZ activity within 48 h of fixation at both 4°C and at room temperature. In conclusion, acetone is the optimal fixative to improve permeability for staining of lacZ activity in large volumes of adult hard tissues

Lipid Composition and Antioxidant Activities of the Seed Oil from Three Malvaceae Species

The oil content and fatty acids, unsaponifiable composition and antioxidant activities of the seed oil from three Malvaceae species (Malva sylvestris L., Malva sylvestris L. var. mauritiana and Althaea officinalis L.) from Serbia were determined. The oil yields from the seeds varied from 7.18 to 9.60%. The main fatty acids of the seed oils were linoleic acid (44.14-54.49%), oleic acid (13.00-16.99%) and palmitic acid (11.45-24.29%). A small amount of cyclopropenoid acids, up to 1.85% was also established. The predominant sterol in all seed oils was beta-sitosterol (11.51-17.34 mg/g of oil). The antioxidant potential of all the investigated seed oils was evaluated by radical scavenging activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay

Autocorrelation of the Ground Vibrations Recorded by the SEIS-InSight Seismometer on Mars

Since early February 2019, the SEIS (Seismic Experiment for Interior Structure) seismometer deployed at the surface of Mars in the framework of the InSight mission has been continuously recording the ground motion at Elysium Planitia. In this study, we take advantage of this exceptional data set to put constraints on the crustal properties of Mars using seismic interferometry (SI). To carry out this task, we first examine the continuous records from the very broadband seismometer. Several deterministic sources of environmental noise are identified and specific preprocessing strategies are presented to mitigate their influence. Applying the principles of SI to the single-station configuration of InSight, we compute, for each Sol and each hour of the martian day, the diagonal elements of the time-domain correlation tensor of random ambient vibrations recorded by SEIS. A similar computation is performed on the diffuse waveforms generated by more than a hundred Marsquakes. A careful signal-to-noise ratio analysis and an inter-comparison between the two datasets suggest that the results from SI are most reliable in a narrow frequency band around 2.4 Hz, where an amplification of both ambient vibrations and seismic events is observed. The average autocorrelation functions (ACFs) contain well identifiable seismic arrivals, that are very consistent between the two datasets. Interpreting the vertical and horizontal ACFs as, respectively, the P- and S- seismic reflectivity below InSight, we propose a simple stratified velocity model of the crust, which is mostly compatible with previous results from receiver function analysis. Our results are discussed and compared to recent works from the literature.This study is InSight contribution number 164. The authors acknowledge both “Université Fédérale de Toulouse Midi Pyrénées” and the “Région Occitanie” for funding the PhD grant of Nicolas Compaire. The French authors acknowledge the French Space Agency CNES and ANR (ANR-14-CE36-0012-02 and ANR-19-CE31-0008-08) for funding the InSight Science analysis

The Far Side of Mars: Two Distant Marsquakes Detected by InSight

For over three Earth years the Marsquake Service has been analyzing the data sent back from the Seismic Experiment for Interior Structure¿the seismometer placed on the surface of Mars by NASA¿s InSight lander. Although by October 2021, the Mars seismic catalog included 951 events, until recently all these events have been assessed as lying within a radius of 100° of InSight. Here we report two distant events that occurred within days of each other, located on the far side of Mars, giving us our first glimpse into Mars¿ core shadow zone. The first event, recorded on 25 August 2021 (InSight sol 976), shows clear polarized arrivals that we interpret to be PP and SS phases at low frequencies and locates to Valles Marineris, 146° ± 7° from InSight. The second event, occurring on 18 September 2021 (sol 1000), has significantly more broadband energy with emergent PP and SS arrivals, and a weak phase arriving before PP that we interpret as Pdiff¿. Considering uncertain pick times and poorly constrained travel times for Pdiff¿, we estimate this event is at a distance between 107° and 147° from InSight. With magnitudes of MMaw 4.2 and 4.1, respectively, these are the largest seismic events recorded so far on Mars.Anna C. Horleston, Jessica C. E. Irving,and Nicholas A. Teanby are funded by the UKSA under Grant Numbers ST/R002096/1, ST/W002523/1, and ST/W002515/1.Nikolaj L. Dahmen, Cecilia Duran, Géraldine Zenhäusern, andSimon C. Stähler would like to acknowledge support from Eidgenössische Technische Hochschule (ETH) through the ETH+ funding scheme (ETH+02 19-1: “Planet Mars”). The French coauthors acknowledge the funding support provided by CNES and the Agence Nationale de la Recherche (ANR-19-CE31-0008-08 MAGIS) for SEIS operation and SEIS Science analysis. Alexander E. Stott acknowledges the French Space Agency CNES and ANR (ANR-19-CE31-0008-08). Caroline Beghein and Jiaqi Li were supported by NASA InSight Participating Scientist Program (PSP) Grant Number 80NSSC18K1679. This article is InSight Contribution Number 236

Seismic Constraints on the Thickness and Structure of the Martian Crust from InSight

NASA¿s InSight mission [1] has for the first time placed a very broad-band seismometer on the surface of Mars. The Seismic Experiment for Interior Structure (SEIS) [2] has been collecting continuous data since early February 2019. The main focus of InSight is to enhance our understanding of the internal structure and dynamics of Mars, which includes the goal to better constrain the crustal thickness of the planet [3]. Knowing the present-day crustal thickness of Mars has important implications for its thermal evolution [4] as well as for the partitioning of silicates and heat-producing elements between the different layers of Mars. Current estimates for the crustal thickness of Mars are based on modeling the relationship between topography and gravity [5,6], but these studies rely on different assumptions, e.g. on the density of the crust and upper mantle, or the bulk silicate composition of the planet and the crust. The resulting values for the average crustal thickness differ by more than 100%, from 30 km to more than 100 km [7]. New independent constraints from InSight will be based on seismically determining the crustal thickness at the landing site. This single firm measurement of crustal thickness at one point on the planet will allow to constrain both the average crustal thickness of Mars as well as thickness variations across the planet when combined with constraints from gravity and topography [8]. Here we describe the determination of the crustal structure and thickness at the InSight landing site based on seismic receiver functions for three marsquakes compared with autocorrelations of InSight data [9].We acknowledge NASA, CNES, partner agencies and institutions (UKSA, SSO,DLR, JPL, IPGP-CNRS, ETHZ, IC, MPS-MPG) and the operators of JPL, SISMOC, MSDS, IRIS-DMC and PDS for providing SEED SEIS data. InSight data is archived in the PDS, and a full list of archives in the Geosciences, Atmospheres, and Imaging nodes is at https://pds-geosciences.wustl.edu/missions/insight/. This work was partially carried out at the Jet Propulsion Laboratory, California Institute of Technology, under a contract with the National Aeronautics and Space Administration. ©2021, California Institute of Technology. Government sponsorship acknowledge

Seismic detection of the martian core

Clues to a planet's geologic history are contained in its interior structure, particularly its core. We detected reflections of seismic waves from the core-mantle boundary of Mars using InSight seismic data and inverted these together with geodetic data to constrain the radius of the liquid metal core to 1830 +/- 40 kilometers. The large core implies a martian mantle mineralogically similar to the terrestrial upper mantle and transition zone but differing from Earth by not having a bridgmanite-dominated lower mantle. We inferred a mean core density of 5.7 to 6.3 grams per cubic centimeter, which requires a substantial complement of light elements dissolved in the iron-nickel core. The seismic core shadow as seen from InSight's location covers half the surface of Mars, including the majority of potentially active regions-e.g., Tharsis-possibly limiting the number of detectable marsquakes.This is InSight contribution 200. We acknowledge NASA, CNES, and partner agencies and institutions (UKSA, SSO, ESA-PRODEX, DLR, JPL, IPGP-CNRS, ETHZ, IC, and MPS-MPG) for the development of SEIS. Numerical simulations were supported by a grant from the Swiss National Supercomputing Centre (CSCS) under project ID s922 as well as HPC resources of CINES under the allocation A0090407341, made by GENCI. We thank B. Dintrans, director of CINES, for his efficient handling of our request for computational time. Figures were created using matplotlib (83), seismic data processing was done in ObsPy (84), and numerical evaluation was done in NumPy and SciPy (85, 86). Funding: S.C.S., A.K., D.G., J.C., A.C.D., G.Z., and N.D. acknowledge support from ETHZ through the ETH+ funding scheme (ETH+2 19-1: “Planet MARS”). S.C.S. acknowledges funding from ETH research grant ETH-10 17-3. W.B.B., A.G.M., M.P.P., and S.E.S. were supported by the NASA InSight mission and funds from the Jet Propulsion Laboratory, California Institute of Technology, under a contract with the National Aeronautics and Space Administration (80NM0018D0004). D.A. has received funding from the European Research Council (ERC) under the European Union’s Horizon 2020 research and innovation program (grant agreement 724690). The French teams acknowledge support from CNES as well as Agence Nationale de la Recherche (ANR-14-CE36-0012-02 and ANR-19-CE31-0008-08). A.R. was financially supported by the Belgian PRODEX program managed by the European Space Agency in collaboration with the Belgian Federal Science Policy Office. M.S. wishes to thank SANIMS (RTI2018-095594-B-I00). M.v.D. received support from the ERC under the European Union’s Horizon 2020 program (grant no. 714069). D.S. and C.S. acknowledge funding from ETH research grant ETH-06 17-02. J.C.E.I. acknowledges support from NASA grant 80NSSC18K1633. N.S., D.K., Q.H., R.M., V.L., and A.G.M. acknowledge NASA grant 80NSSC18K1628 for support. V.L. acknowledges support from the Packard Foundation. W.T.P. and C.C. received funding from the UK Space Agency, grant ST/S001239/1. A.H. was funded by the UK Space Agency (grant ST/R002096/1). A.-C.P. acknowledges the financial support and endorsement from the DLR Management Board Young Research Group Leader Program and the Executive Board Member for Space Research and Technology. Author contributions: S.C.S., D.G., S.C., R.F.G., Q.H., D.K., V.L., M.S., N.S., D.S., É.S., C.S., and G.Z. analyzed the seismic data and made ScS arrival time picks. S.C.S., P.L., D.G., Z.X., C.C., and W.T.P. performed the statistical analysis of the observed signals. S.C.S., Q.H., N.S., R.M., and A.G.M. identified the arrivals as ScS waves based on interior models from A.K., H.S., and A.R. A.K., M.D., A.C.D., and H.S. performed the inversions. S.C.S., A.K., P.L., D.G., D.A., J.C.E.I., M.K., C.P., A.-C.P., A.R., T.G., and S.E.S. participated and contributed to the interpretation of the results. Review of the continuous data and detection of marsquakes was done by S.C.S., S.C., G.Z., C.C., N.D., J.C., M.v.D., T.K., M.P., and A.H. with operational support by É.B., C.P., and P.M.D. S.C.S. and A.K. wrote the central part of the paper with contributions from H.S., N.S., D.A., J.C.E.I., A.G.M., A.-C.P., A.R., J.C., and M.v.D. J.C.E.I., R.M., M.K., and V.L. reviewed the contributions to the supplementary materials. The InSight mission is managed by W.B.B., M.P.P., and S.E.S. The SEIS instrument development was led by P.L., D.G., W.T.P., and W.B.B. Supplementary section 1 was written by M.S., D.S., and É.S. with contributions from S.C.S., C.S., and Z.X. Supplementary section 2 was written by D.K. and V.L. with contributions from J.C.E.I. and N.S. Supplementary section 3 was written by M.S. and É.S. Supplementary section 4 was written by R.F.G. with contributions from M.D. Supplementary section 5 was written by Q.H. with contributions from N.S. Supplementary section 6 was written by S.C.S. with contributions from the authors of the other supplements. Supplementary section 7 was written by Z.X. and C.C. with contributions from P.L. and W.T.P. Supplementary section 8 was written by A.K., M.D., A.C.D., and H.S. Supplementary section 9 was written by M.D. Supplementary section 10 was written by A.C.D., A.K., and M.D. Supplementary section 11 was written by D.A. and A.R. with contributions from A.K. Competing interests: The authors declare that they have no competing interests. Data and materials availability: We thank the operators of JPL, SISMOC, MSDS, IRIS-DMC, and PDS for providing SEED SEIS data (87). Three hundred interior models derived in this study are available from MSDS (88)

Seismic detection of the martian core by InSight

A plethora of geophysical, geo- chemical, and geodynamical observations indicate that the terrestrial planets have differentiated into silicate crusts and mantles that surround a dense core. The latter consists primarily of Fe and some lighter alloying elements (e.g., S, Si, C, O, and H) [1]¿. The Martian meteorites show evidence of chalcophile element depletion, suggesting that the otherwise Fe-Ni- rich core likely contains a sulfide component, which influences physical state

Surface waves and crustal structure on Mars

We detected surface waves from two meteorite impacts on Mars. By measuring group velocity dispersion along the impact-lander path, we obtained a direct constraint on crustal structure away from the InSight lander. The crust north of the equatorial dichotomy had a shear wave velocity of approximately 3.2 kilometers per second in the 5- to 30-kilometer depth range, with little depth variation. This implies a higher crustal density than inferred beneath the lander, suggesting either compositional differences or reduced porosity in the volcanic areas traversed by the surface waves. The lower velocities and the crustal layering observed beneath the landing site down to a 10-kilometer depth are not a global feature. Structural variations revealed by surface waves hold implications for models of the formation and thickness of the martian crust.D.K., S.C., D.G., J.C., C.D., A. K., S.C.S., N.D., and G.Z. were supported by the ETH+ funding scheme (ETH+02 19-1: “Planet Mars”). Marsquake Service operations at ETH Zürich were supported by ETH Research grant ETH-06 17-02. N.C.S. and V.L. were supported by NASA PSP grant no. 80NSSC18K1628. Q.H. and E.B. are funded by NASA grant 80NSSC18K1680. C.B. and J.L. were supported by NASA InSight PSP grant no. 80NSSC18K1679. S.D.K. was supported by NASA InSight PSP grant no. 80NSSC18K1623. P.L., E.B., M.D., H.S., E.S., M.W., Z.X., T.W., M.P., R.F.G. were supported by CNES and the Agence Nationale de la Recherche (ANR-19-CE31-0008-08 MAGIS) for SEIS operation and SEIS Science analysis. A.H., C.C. and W.T.P. were supported by the UKSA under grant nos. ST/R002096/1, ST/ W002523/1 and ST/V00638X/1. Numerical computations of McMC Approach 2 were performed on the S-CAPAD/DANTE platform (IPGP, France) and using the HPC resources of IDRIS under the allocation A0110413017 made by GENCI. A.H. was supported by the UKSA under grant nos. ST/R002096/1 and ST/W002523/1. F.N. was supported by InSight PSP 80NSSC18K1627. I.J.D. was supported by NASA InSight PSP grant no. 80NSSC20K0971. L.V.P. was funded by NASANNN12AA01C with subcontract JPL-1515835. The research was carried out in part by W.B.B., M.G. and M.P.P. at the Jet Propulsion Laboratory, California Institute of Technology, under a contract with the National Aeronautics and Space Administration (80NM0018D0004)Peer reviewe

The role of mechanotransduction versus hypoxia during simulated orthodontic compressive strain—an in vitro study of human periodontal ligament fibroblasts

During orthodontic tooth movement (OTM) mechanical forces trigger pseudo-inflammatory, osteoclastogenic and remodelling processes in the periodontal ligament (PDL) that are mediated by PDL fibroblasts via the expression of various signalling molecules. Thus far, it is unknown whether these processes are mainly induced by mechanical cellular deformation (mechanotransduction) or by concomitant hypoxic conditions via the compression of periodontal blood vessels. Human primary PDL fibroblasts were randomly seeded in conventional six-well cell culture plates with O-2-impermeable polystyrene membranes and in special plates with gas-permeable membranes (Lumox (R), Sarstedt), enabling the experimental separation of mechanotransducive and hypoxic effects that occur concomitantly during OTM. To simulate physiological orthodontic compressive forces, PDL fibroblasts were stimulated mechanically at 2 g.cm(-2) for 48 h after 24 h of pre-incubation. We quantified the cell viability by MTT assay, gene expression by quantitative real-time polymerase chain reaction (RT-qPCR) and protein expression by western blot/enzyme-linked immunosorbent assays (ELISA). In addition, PDL-fibroblast-mediated osteoclastogenesis (TRAP(+) cells) was measured in a 72-h coculture with RAW264.7 cells. The expression of HIF-1 alpha, COX-2, PGE2, VEGF, COL1A2, collagen and ALPL, and the RANKL/OPG ratios at the mRNA/protein levels during PDL-fibroblast-mediated osteoclastogenesis were significantly elevated by mechanical loading irrespective of the oxygen supply, whereas hypoxic conditions had no significant additional effects. The cellular-molecular mediation of OTM by PDL fibroblasts via the expression of various signalling molecules is expected to be predominantly controlled by the application of force (mechanotransduction), whereas hypoxic effects seem to play only a minor role. In the context of OTM, the hypoxic marker HIF-1 alpha does not appear to be primarily stabilized by a reduced O-2 supply but is rather stabilised mechanically