Assessment of soil organic matter supply: Challenges and opportunities

Soil organic matter (SOM) is recognized as an important factor for sustainable land use. Several analyzing techniques were focused on fractionation of soil organic carbon (SOC), on carbon sequestration, soil functions, or other approaches. We combined SOC fractionation with studies on carbon sequestration. Thermogravimetry (TG, recording of mass losses during heating up of soil samples) was selected as a supplemental method to standard analyzing techniques for soils. TG provides recording of thermal mass losses in dependency on temperature what facilitate fractionation together with SOM content determination via mass losses on ignition (MLI). Autocorrelation analyses of TG data enable to assess the carbon sequestration processes. After a gentle sample preparation, more than 370 soil samples in eight sample sets were analyzed from different types of soils and regions of origin. The results extend literature data by revealing quantifiable interrelations between content of SOM, SOC and clay with a coefficient of determination around 0.98. Deviations from the relationship become lower during incubation experiments, with increasing sampling depth, and with decreasing organic fertilization in plots of long term agriculture field experiments etc. We explained these results with changing quantities of extraneous (mostly fresh) organic residues not affected by soil carbon content regulation. These organic residues seems to be quantifiable via difference between measured MLI and the MLI calculated from content of SOC and clay both determined by standard methods. The practical use of found interrelation implies an acceptance of traditional definition of soil and SOM as products of long term ecosystem succession with content regulation as a unifying over regions soil feature. In contrast, the more common and simplified understanding of soil as carbon containing mineral substrates supports public recognition of soils. However, it does not facilitate the comparison of results from different regions and studies about soils. We conclude from these considerations about obligatory distinction between following types of organic carbon as an essential precondition for assessment of SOM supply: 1. SOC (or humus) as a product of long term carbon regulation processes, 2. fresh organic residues, and carbon of 3. geologic (turf, coal, graphite, diamond, …) or 4. anthropogenic origin (black carbon in ashes, cinder, soot, asphalt)

Organic carbon content determination in soils: challenges and opportunities of elemental analysis versus thermogravimetry

Sustainable soil management needs reliable and accurate monitoring of soil organic carbon (SOC) content. However, despite of the development of analytical techniques during last decades, the detection opportunities for short term and rather small changes in SOC induced by organic fertilization, organic amendments or land use changes are still limited with the available methods. This study aims to quantify the theoretical detection opportunities for changes in SOC content with elemental analysis (EA) as the standard method in comparing with thermogravimetry (TG) as an enhanced traditional approach derived from soil organic matter determination via mass losses on ignition. The carried out experiments consist of mixing soil samples from non-fertilized plots of three long-term agricultural experiments in Bad Lauchstaedt, Großbeeren and Muencheberg (silty loam, loamy sand and silty sand) with straw, farmyard manure, sheep faeces and charcoal in four quantities (3 t×ha-1, 20 t×ha-1, 60 t×ha-1 and 180 t×ha‑1fresh matter) under laboratory conditions.The quantities were based on fresh matter application in agricultural practice accepting different amounts of added organic carbon. The results confirm EA as a method of higher reliability and accuracy for carbon content determination. TG allows to distinguish the different types of added amendments with high sensitivity. This was achieved by using newly developed evaluation algorithms for the thermal decay dynamics. We conclude from these results that TG cannot substitute EA to determine organic carbon on a routine base. However, TG could be a supplementary fingerprinting technique for the detection of added organic carbon to soils from organic fertilizers and to distinguish sources of geological or anthropogenic origin enabling a future assessment of soil organic carbon quality

Novel insights into host-fungal pathogen interactions derived from live-cell imaging

Acknowledgments The authors acknowledge funding from the Wellcome Trust (080088, 086827, 075470 and 099215) including a Wellcome Trust Strategic Award for Medical Mycology and Fungal Immunology 097377 and FP7-2007–2013 grant agreement HEALTH-F2-2010-260338–ALLFUN to NARG.Peer reviewedPublisher PD

Inhibiting ex-vivo Th17 responses in Ankylosing Spondylitis by targeting Janus kinases

Treatment options for Ankylosing Spondylitis (AS) are still limited. The T helper cell 17 (Th17) pathway has emerged as a major driver of disease pathogenesis and a good treatment target. Janus kinases (JAK) are key transducers of cytokine signals in Th17 cells and therefore promising targets for the treatment of AS. Here we investigate the therapeutic potential of four different JAK inhibitors on cells derived from AS patients and healthy controls, cultured in-vitro under Th17-promoting conditions. Levels of IL-17A, IL-17F, IL-22, GM-CSF and IFN gamma were assessed by ELISA and inhibitory effects were investigated with Phosphoflow. JAK1/2/3 and TYK2 were silenced in CD4+ T cells with siRNA and effects analyzed by ELISA (IL-17A, IL-17F and IL-22), Western Blot, qPCR and Phosphoflow. In-vitro inhibition of CD4+ T lymphocyte production of multiple Th17 cytokines (IL-17A, IL-17F and IL-22) was achieved with JAK inhibitors of differing specificity, as well as by silencing of JAK1-3 and Tyk2, without impacting on cell viability or proliferation. Our preclinical data suggest JAK inhibitors as promising candidates for therapeutic trials in AS, since they can inhibit multiple Th17 cytokines simultaneously. Improved targeting of TYK2 or other JAK isoforms may confer tailored effects on Th17 responses in AS

A Sulfhydryl-Reactive Ruthenium (II) Complex and Its Conjugation to Protein G as a Universal Reagent for Fluorescent Immunoassays

To develop a fluorescent ruthenium complex for biosensing, we synthesized a novel sulfhydryl-reactive compound, 4-bromophenanthroline bis-2,2′-dipyridine Ruthenium bis (hexafluorophosphate). The synthesized Ru(II) complex was crosslinked with thiol-modified protein G to form a universal reagent for fluorescent immunoassays. The resulting Ru(II)-protein G conjugates were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The emission peak wavelength of the Ru(II)-protein G conjugate was 602 nm at the excitation of 452 nm which is similar to the spectra of the Ru(II) complex, indicating that Ru(II)-protein G conjugates still remain the same fluorescence after conjugation. To test the usefulness of the conjugate for biosensing, immunoglobulin G (IgG) binding assay was conducted. The result showed that Ru(II)-protein G conjugates were capable of binding IgG and the more cross-linkers to modify protein G, the higher conjugation efficiency. To demonstrate the feasibility of Ru(II)-protein G conjugates for fluorescent immunoassays, the detection of recombinant histidine-tagged protein using the conjugates and anti-histidine antibody was developed. The results showed that the histidine-tagged protein was successfully detected with dose-response, indicating that Ru(II)-protein G conjugate is a useful universal fluorescent reagent for quantitative immunoassays

Identification of Milk Component in Ancient Food Residue by Proteomics

Proteomic approaches based on mass spectrometry have been recently used in archaeological and art researches, generating promising results for protein identification. Little information is known about eastward spread and eastern limits of prehistoric milking in eastern Eurasia.In this paper, an ancient visible food remain from Subeixi Cemeteries (cal. 500 to 300 years BC) of the Turpan Basin in Xinjiang, China, preliminarily determined containing 0.432 mg/kg cattle casein with ELISA, was analyzed by using an improved method based on liquid chromatography (LC) coupled with MALDI-TOF/TOF-MS to further identify protein origin. The specific sequence of bovine casein and the homology sequence of goat/sheep casein were identified.The existence of milk component in ancient food implies goat/sheep and cattle milking in ancient Subeixi region, the furthest eastern location of prehistoric milking in the Old World up to date. It is envisioned that this work provides a new approach for ancient residue analysis and other archaeometry field

Crystal Structures of ABL-Related Gene (ABL2) in Complex with Imatinib, Tozasertib (VX-680), and a Type I Inhibitor of the Triazole Carbothioamide Class†

ABL2 (also known as ARG (ABL related gene)) is closely related to the well-studied Abelson kinase cABL. ABL2 is involved in human neoplastic diseases and is deregulated in solid tumors. Oncogenic gene translocations occur in acute leukemia. So far no structural information for ABL2 has been reported. To elucidate structural determinants for inhibitor interaction, we determined the cocrystal structure of ABL2 with the oncology drug imatinib. Interestingly, imatinib not only interacted with the ATP binding site of the inactive kinase but was also bound to the regulatory myristate binding site. This structure may therefore serve as a tool for the development of allosteric ABL inhibitors. In addition, we determined the structures of ABL2 in complex with VX-680 and with an ATP-mimetic type I inhibitor, which revealed an interesting position of the DFG motif intermediate between active and inactive conformations, that may also serve as a template for future inhibitor design

Allosteric Interactions between the Myristate- and ATP-Site of the Abl Kinase

Abl kinase inhibitors targeting the ATP binding pocket are currently employed as potent anti-leukemogenic agents but drug resistance has become a significant clinical limitation. Recently, a compound that binds to the myristate pocket of Abl (GNF-5) was shown to act cooperatively with nilotinib, an ATP-competitive inhibitor to target the recalcitrant “T315I” gatekeeper mutant of Bcr-Abl. To uncover an explanation for how drug binding at a distance from the kinase active site could lead to inhibition and how inhibitors could combine their effects, hydrogen exchange mass spectrometry (HX MS) was employed to monitor conformational effects in the presence of both dasatinib, a clinically approved ATP-site inhibitor, and GNF-5. While dasatinib binding to wild type Abl clearly influenced Abl conformation, no binding was detected between dasatinib and T315I. GNF-5, however, elicited the same conformational changes in both wild type and T315I, including changes to dynamics within the ATP site located approximately 25 Å from the site of GNF-5 interaction. Simultaneous binding of dasatinib and GNF-5 to T315I caused conformational and/or dynamics changes in Abl such that effects of dasatinib on T315I were the same as when it bound to wild type Abl. These results provide strong biophysical evidence that allosteric interactions play a role in Abl kinase downregulation and that targeting sites outside the ATP binding site can provide an important pharmacological tool to overcome mutations that cause resistance to ATP-competitive inhibitors