Dental pulp pluripotent-like stem cells (DPPSC), a new stem cell population with chromosomal stability and osteogenic capacity for biomaterials evaluation

Background: Biomaterials are widely used to regenerate or substitute bone tissue. In order to evaluate their potential use for clinical applications, these need to be tested and evaluated in vitro with cell culture models. Frequently, immortalized osteoblastic cell lines are used in these studies. However, their uncontrolled proliferation rate, phenotypic changes or aberrations in mitotic processes limits their use in long-term investigations. Recently, we described a new pluripotent-like subpopulation of dental pulp stem cells derived from the third molars (DPPSC) that shows genetic stability and shares some pluripotent characteristics with embryonic stem cells. In this study we aim to describe the use of DPPSC to test biomaterials, since we believe that the biomaterial cues will be more critical in order to enhance the differentiation of pluripotent stem cells. Methods: The capacity of DPPSC to differentiate into osteogenic lineage was compared with human sarcoma osteogenic cell line (SAOS-2). Collagen and titanium were used to assess the cell behavior in commonly used biomaterials. The analyses were performed by flow cytometry, alkaline phosphatase and mineralization stains, RT-PCR, immunohistochemistry, scanning electron microscopy, Western blot and enzymatic activity. Moreover, the genetic stability was evaluated and compared before and after differentiation by short-comparative genomic hybridization (sCGH). Results: DPPSC showed excellent differentiation into osteogenic lineages expressing bone-related markers similar to SAOS-2. When cells were cultured on biomaterials, DPPSC showed higher initial adhesion levels. Nevertheless, their osteogenic differentiation showed similar trend among both cell types. Interestingly, only DPPSC maintained a normal chromosomal dosage before and after differentiation on 2D monolayer and on biomaterials. Conclusions: Taken together, these results promote the use of DPPSC as a new pluripotent-like cell model to evaluate the biocompatibility and the differentiation capacity of biomaterials used in bone regeneration

Rapid Intensity Decrease During the Second Half of the First Millennium BCE in Central Asia and Global Implications

Recent paleomagnetic studies have shown that important short-lived intensity fluctuations occurred during the first millennium BCE. However, the knowledge of the spatial and temporal extension of these features is still limited by the scarce availability of robust data. In this study we focus on the study of the intensity decrease that took place in Central Asia during the second half of the 1st millennium BCE after the high intensities that characterized the Levantine Iron Age Anomaly. Since previous archeointensities available for this period and region were obtained without accomplishing modern standards of quality, we present here new archeointensities that are derived from classical Thellier and Thellier experiments, including partial thermoremanent magnetization (pTRM) checks, thermoremanent magnetization (TRM) anisotropy and cooling rate corrections at the specimen level. The new 51 archeointensities, together with previous archeointensities, have been used to present a new local paleosecular variation curve for Central Asia. The results confirm the existence of an important geomagnetic field intensity decrease in South Uzbekistan from the 4th century BCE to the end of the 1st century BCE associated with rates of changes up to −15 μT/century. A critical analysis of the archeointensity global database indicates that this feature was present at continental scale, from Western Europe to Central Asia. However, this trend is not identified in other regions such as Japan or Mexico. Finally, the comparison with the dipole moment derived from recent global geomagnetic field reconstructions suggests a strong influence of non-dipolar sources upon this continental intensity feature

Mapping the ionized gas of the metal-poor HII galaxy PHL 293B with MEGARA

Here we report the first spatially resolved spectroscopic study for the galaxy PHL293B using the high-resolution GTC/MEGARA IFU. PHL293B is a local, extremely metal-poor, high ionization galaxy. This makes PHL 293B an excellent analogue for galaxies in the early Universe. The MEGARA aperture (~12.5''x 11.3'') covers the entire PHL 293B main body and its far-reaching ionized gas. We created and discussed maps of all relevant emission lines, line ratios and physical-chemical properties of the ionized ISM. The narrow emission gas appears to be ionized mainly by massive stars according to the observed diganostic line ratios, regardless of the position across the MEGARA aperture. We detected low intensity broad emission components and blueshifted absorptions in the Balmer lines (H$\alpha$,H$\beta$) which are located in the brightest zone of the galaxy ISM. A chemically homogeneity, across hundreds of parsecs, is observed in O/H. We take the oxygen abundance 12+log(O/H)=7.64 $\pm$ 0.06 derived from the PHL293B integrated spectrum as the representative metallicity for the galaxy. Our IFU data reveal for the first time that the nebular HeII4686 emission from PHL 293B is spatially extended and coincident with the ionizing stellar cluster, and allow us to compute its absolute HeII ionizing photon flux. Wolf-Rayet bumps are not detected excluding therefore Wolf-Rayet stars as the main HeII excitation source. The origin of the nebular HeII4686 is discussed.Comment: 14 pages, 9 Figures, 3 Tables; Accepted for publication in MNRA

Artificial intelligence-based software (AID-FOREST) for tree detection: A new framework for fast and accurate forest inventorying using LiDAR point clouds

Forest inventories are essential to accurately estimate different dendrometric and forest stand parameters. However, classical forest inventories are time consuming, slow to conduct, sometimes inaccurate and costly. To address this problem, an efficient alternative approach has been sought and designed that will make this type of field work cheaper, faster, more accurate, and easier to complete. The implementation of this concept has required the development of a specifically designed software called "Artificial Intelligence for Digital Forest (AID-FOREST)", which is able to process point clouds obtained via mobile terrestrial laser scanning (MTLS) and then, to provide an array of multiple useful and accurate dendrometric and forest stand parameters. Singular characteristics of this approach are: No data pre-processing is required either pre-treatment of forest stand; fully automatic process once launched; no limitations by the size of the point cloud file and fast computations.To validate AID-FOREST, results provided by this software were compared against the obtained from in-situ classical forest inventories. To guaranty the soundness and generality of the comparison, different tree spe-cies, plot sizes, and tree densities were measured and analysed. A total of 76 plots (10,887 trees) were selected to conduct both a classic forest inventory reference method and a MTLS (ZEB-HORIZON, Geoslam, ltd.) scanning to obtain point clouds for AID-FOREST processing, known as the MTLS-AIDFOREST method. Thus, we compared the data collected by both methods estimating the average number of trees and diameter at breast height (DBH) for each plot. Moreover, 71 additional individual trees were scanned with MTLS and processed by AID-FOREST and were then felled and divided into logs measuring 1 m in length. This allowed us to accurately measure the DBH, total height, and total volume of the stems.When we compared the results obtained with each methodology, the mean detectability was 97% and ranged from 81.3 to 100%, with a bias (underestimation by MTLS-AIDFOREST method) in the number of trees per plot of 2.8% and a relative root-mean-square error (RMSE) of 9.2%. Species, plot size, and tree density did not significantly affect detectability. However, this parameter was significantly affected by the ecosystem visual complexity index (EVCI). The average DBH per plot was underestimated (but was not significantly different from 0) by the MTLS-AIDFOREST, with the average bias for pooled data being 1.8% with a RMSE of 7.5%. Similarly, there was no statistically significant differences between the two distribution functions of the DBH at the 95.0% confidence level.Regarding the individual tree parameters, MTLS-AIDFOREST underestimated DBH by 0.16 % (RMSE = 5.2 %) and overestimated the stem volume (Vt) by 1.37 % (RMSE = 14.3 %, although the BIAS was not statistically significantly different from 0). However, the MTLS-AIDFOREST method overestimated the total height (Ht) of the trees by a mean 1.33 m (5.1 %; relative RMSE = 11.5 %), because of the different height concepts measured by both methodological approaches. Finally, AID-FOREST required 30 to 66 min per ha-1 to fully automatically process the point cloud data from the *.las file corresponding to a given hectare plot. Thus, applying our MTLS-AIDFOREST methodology to make full forest inventories, required a 57.3 % of the time required to perform classical plot forest inventories (excluding the data postprocessing time in the latter case). A free trial of AID -FOREST can be requested at [email protected]

Sarcoptic mange in wild ruminants in Spain: solving the epidemiological enigma using microsatellite markers

Background: In Spain, sarcoptic mange was first described in native wildlife in 1987 in Cazorla Natural Park, causing the death of nearly 95% of the local native population of Iberian ibex (Capra pyrenaica). Since then, additional outbreaks have been identified in several populations of ibex and other wild ungulate species throughout the country. Although the first epizootic outbreak in wildlife was attributed to the introduction of an infected herd of domestic goats, the origin and the cause of its persistence remain unclear. The main aims of this study are to understand (i) the number of Sarcoptes scabiei “strains” circulating in wild ruminant populations in Spain, and (ii) the molecular epidemiological relationships between S. scabiei and its hosts. Methods: Ten Sarcoptes microsatellite markers were used to characterize the genetic structure of 266 mites obtained from skin scrapings of 121 mangy wild ruminants between 2011 and 2019 from 11 areas in Spain. Results: Seventy-three different alleles and 37 private alleles were detected. The results of this study show the existence of three genetic strains of S. scabiei in the wild ruminant populations investigated. While two genetic clusters of S. scabiei were host- and geography-related, one cluster included multi-host mites deriving from geographically distant populations. Conclusions: The molecular epidemiological study of S. scabiei in wild ruminants in Spain indicates that the spreading and persistence of the parasite may be conditioned by host species community composition and the permissiveness of each host population/community to the circulation of individual “strains,” among other factors. Wildlife–livestock interactions and the role of human-driven introduction or trade of wild and domestic animals should be better investigated to prevent further spread of sarcoptic mange in as yet unaffected natural areas of the Iberian Peninsula

Resolving Structure and Mechanical Properties at the Nanoscale of Viruses with Frequency Modulation Atomic Force Microscopy

Structural Biology (SB) techniques are particularly successful in solving virus structures. Taking advantage of the symmetries, a heavy averaging on the data of a large number of specimens, results in an accurate determination of the structure of the sample. However, these techniques do not provide true single molecule information of viruses in physiological conditions. To answer many fundamental questions about the quickly expanding physical virology it is important to develop techniques with the capability to reach nanometer scale resolution on both structure and physical properties of individual molecules in physiological conditions. Atomic force microscopy (AFM) fulfills these requirements providing images of individual virus particles under physiological conditions, along with the characterization of a variety of properties including local adhesion and elasticity. Using conventional AFM modes is easy to obtain molecular resolved images on flat samples, such as the purple membrane, or large viruses as the Giant Mimivirus. On the contrary, small virus particles (25–50 nm) cannot be easily imaged. In this work we present Frequency Modulation atomic force microscopy (FM-AFM) working in physiological conditions as an accurate and powerful technique to study virus particles. Our interpretation of the so called “dissipation channel” in terms of mechanical properties allows us to provide maps where the local stiffness of the virus particles are resolved with nanometer resolution. FM-AFM can be considered as a non invasive technique since, as we demonstrate in our experiments, we are able to sense forces down to 20 pN. The methodology reported here is of general interest since it can be applied to a large number of biological samples. In particular, the importance of mechanical interactions is a hot topic in different aspects of biotechnology ranging from protein folding to stem cells differentiation where conventional AFM modes are already being used

Nivolumab and sunitinib combination in advanced soft tissue sarcomas : A multicenter, single-arm, phase Ib/II trial

Sarcomas exhibit low expression of factors related to immune response, which could explain the modest activity of PD-1 inhibitors. A potential strategy to convert a cold into an inflamed microenvironment lies on a combination therapy. As tumor angiogenesis promotes immunosuppression, we designed a phase Ib/II trial to test the double inhibition of angiogenesis (sunitinib) and PD-1/PD-L1 axis (nivolumab). This single-arm, phase Ib/II trial enrolled adult patients with selected subtypes of sarcoma. Phase Ib established two dose levels: level 0 with sunitinib 37.5 mg daily from day 1, plus nivolumab 3 mg/kg intravenously on day 15, and then every 2 weeks; and level-1 with sunitinib 37.5 mg on the first 14 days (induction) and then 25 mg per day plus nivolumab on the same schedule. The primary endpoint was to determine the recommended dose for phase II (phase I) and the 6-month progression-free survival rate, according to Response Evaluation Criteria in Solid Tumors 1.1 (phase II). From May 2017 to April 2019, 68 patients were enrolled: 16 in phase Ib and 52 in phase II. The recommended dose of sunitinib for phase II was 37.5 mg as induction and then 25 mg in combination with nivolumab. After a median follow-up of 17 months (4-26), the 6-month progression-free survival rate was 48% (95% CI 41% to 55%). The most common grade 3-4 adverse events included transaminitis (17.3%) and neutropenia (11.5%). Sunitinib plus nivolumab is an active scheme with manageable toxicity in the treatment of selected patients with advanced soft tissue sarcoma, with almost half of patients free of progression at 6 months

A search for charged massive long-lived particles

We report on a search for charged massive long-lived particles (CMLLPs), based on 5.2 fb$^{-1}$ of integrated luminosity collected with the D0 detector at the Fermilab Tevatron $p\bar{p}$ collider. We search for events in which one or more particles are reconstructed as muons but have speed and ionization energy loss $(dE/dx)$ inconsistent with muons produced in beam collisions. CMLLPs are predicted in several theories of physics beyond the standard model. We exclude pair-produced long-lived gaugino-like charginos below 267 GeV and higgsino-like charginos below 217 GeV at 95% C.L., as well as long-lived scalar top quarks with mass below 285 GeV.Comment: submitted to Phys. Rev. Letter