Combination of Trabectedin and Gemcitabine for Advanced Soft Tissue Sarcomas: Results of a Phase I Dose Escalating Trial of the German Interdisciplinary Sarcoma Group (GISG)

Background: Evaluation of the potential efficacy and safety of combination therapies for advanced soft tissue sarcomas (STS) has increased substantially after approval of trabectedin and pazopanib. Trabectedin’s introduction in Europe in 2007 depended mainly on its activity in so-called L-sarcomas (liposarcoma and leiomyosarcoma); combination of trabectedin with other chemotherapies used in STS seems of particular interest. Methods: We initiated within the German Interdisciplinary Sarcoma Group (GISG) a phase I dose escalating trial evaluating the combination of trabectedin and gemcitabine in patients with advanced and/or metastatic L-sarcomas (GISG-02; ClinicalTrials.gov {"type":"clinical-trial","attrs":{"text":"NCT01426633","term_id":"NCT01426633"}}NCT01426633). Patients were treated with increasing doses of trabectedin and gemcitabine. The primary endpoint was to determine the maximum tolerated dose. Results: Five patients were included in the study. Two patients were treated on dose level 1 comprising trabectedin 0.9 mg/m² on day 1 and gemcitabine 700 mg/m² on days 1 + 8, every 3 weeks. Due to dose-limiting toxicity (DLT) in both patients (elevated transaminases and thrombocytopenia), an additional three patients were treated on dose level −1 with trabectedin 0.7 mg/m² plus gemcitabine 700 mg/m². Of these three patients, two demonstrated another DLT; therefore, the trial was stopped and none of the dose levels could be recommended for phase II testing. Conclusion: The GISG-02 phase I study was stopped with the conclusion that the combination of gemcitabine and trabectedin is generally not recommended for the treatment of patients with advanced and/or metastatic leiomyosarcoma or liposarcoma. Also, this phase I study strongly supports the necessity for careful evaluation of combination therapies.     

Root Exudation of Organic Acids of Herbaceous Pioneer Plants and Their Growth in Sterile and Non-Sterile Nutrient-Poor, Sandy Soils from Post-Mining Sites

Nutrient-poor, sandy soils form the prevailing substrate at post-mining sites of the Lusatian region(Brandenburg, Germany) and present a challenge for vegetation development. We studied the organic acid quantity and composition of three commonly occurring pioneer plant species, the legumes Lotus corniculatus L. and Trifolium arvense L. and the grass Calamagrostis epigeios(L.) Roth, to determine if plant growth and exudation differed with(non-sterilized soil) and without(sterilized soil) an indigenous soil microbial community. We investigated whether organic acids were found in the rhizosphere and surrounding soil and whether this influenced nutrient mobilization. This study consists of linked field investigations and a greenhouse experiment. Plants were grown in the greenhouse in either sterilized or non-sterilized sandy soil from a reclamation site in the Lusatian mining landscape(Welzow Su¨d, East Germany). After seven months, the plant biomass, root morphology, organic acids, and water-soluble nutrients and root colonization with arbuscular mycorrhizal fungi(AMF) and dark septate endophytes(DSE) were analyzed. Roots of all three plants in the field and greenhouse experiments were highly colonized with AMF. Calamagrostis epigeios and T. arvense had a significantly higher colonization frequency with DSE than L. corniculatus. The quantity and composition of organic acids strongly differed among plant species, with the highest number of organic acids found for L. corniculatus and lowest for C. epigeios. The quantity of organic acids was greatly reduced in all plants under sterilized soil conditions. However, the composition of organic acids and plant growth in sterilized soil were reduced for both legumes, but not for C. epigeios, which had a higher biomass under sterilized conditions. Changes in nutrient concentrations in the field rhizosphere soil relative to those in the control were measurable after seven months. While the spectrum of organic acids and the growth of legumes seemed to be dependent on a highly diverse soil microbial community and a symbiotic partner, the grass C. epigeios appeared capable of mobilizing enough nutrients without an indigenous microbial community, and might be more competitive on sites where soil microbial diversity and activity are low.     

Influence of drying vs. freezing of archived soil samples on soil organic matter fractions

Archived soil samples are a valuable tool for any long‐term soil research. We analysed total carbon (C) and nitrogen (N) content and soil organic matter fractions in 38 archived soil samples that were stored for up to 21 years and compared air‐dried storage to frozen storage conditions. Samples include top‐ and upper subsoils, different soil texture and land use with C contents between 4.3 and 174 mg g^?1. The results from this study reveal no changes in total C and N contents with storage time up to 21 years or type of storage (freezing vs. air drying). The analyses of soil physical fractions also revealed no significant differences between air‐dried stored and frozen stored samples for most samples. However, we found indications, that freezing of soil material might lead to changes in the mineral fractions for soils containing high amounts of water. Therefore, and as archiving soils in a frozen state is more expensive than storing air‐dried samples, we recommend the use of air‐dried samples for C quality analyses of archived soil samples.     

The paramount influence of nitrate in increasing apoplastic pH of young sunflower leaves to induce Fe deficiency chlorosis,and the re-greening effect brought about by acidic foliar sprays

The main objective of the present work was to clarify the causal relationship between leaf apoplastic pH increase and Fe chlorosis under alkaline growth conditions. It has been shown that nitrate supply in contrast to ammonium supply induced a pH increase in the apoplast of young green leaves of Helianthus annuus which was followed within 12 hours by leaf yellowing. Hence nitrate nutrition is the primary cause of a high leaf apoplastic pH which induces Fe deficiency chlorosis and not the impaired provision of ATP for plasmalemma H⁺ pumps in yellow leaves. Supply of bicarbonate in physiological concentrations had virtually no influence on leaf apoplastic pH. Spraying leaves with diluted acids (citric acid, sulphuric acid) resulted in a decrease of apoplastic pH followed by leaf re-greening. Interestingly, the Fe concentrations remained the same in the yellow control leaves and in the sprayed green leaves. From this it follows that Fe efficiency in leaves is mainly related to the Fe distribution between apoplast and symplast. It was demonstrated that Fe chlorosis induced by nitrate nutrition begins from the base of the youngest leaves, presumably from growing interveinal microsites showing high nitrate uptake rates. Leaf yellowing spread gradually from the leaf base to the tip and after seven days of nitrate supply the leaf was almost completely yellow (98%). Leaf yellowing was measured by means of a video imaging technique. Leaf apoplastic pH recordings were conducted after loading the fluorescent dye FITC-Dextran (4000 D) into the leaf apoplast of intact plants thus simulating in vivo conditions. It was also shown using the new loading technique that the fluorescent dye did not penetrate the leaf symplast.     

Correlated effects of exotic pollen‐fertility restorer genes on agronomic and quality traits of hybrid rye

In CMS (cytoplasmic male sterility)‐based hybrid rye (Secale cereale L.) breeding, effective pollen‐fertility restoration is an essential prerequisite for achieving maximum grain yield on the one hand and for minimizing ergot (Claviceps purpurea) infestation on the other. Restorer genes for the CMS‐inducing ‘Pampa’ cytoplasm derived from landraces collected in Iran and Argentina are used by breeders for achieving this goal. Here, restorer genes from four germplasm sources (‘Altevogt 14160’, ‘IRAN III’, ‘Trenelense’ and ‘Pico Gentario’) were analysed by producing three‐way cross hybrids between an elite CMS single cross and pollinators with and without a given restorer gene. Materials were evaluated on large drilled plots for restorer index (RI), grain yield, plant height and other traits in six environments. In experiment 1, a restorer gene from ‘Altevogt 14160’ was used. Seven pairs of marker‐selected carrier and non‐carrier backcross lines served as pollinators. In experiment 2, the pollinators were 17 backcross line pairs from the other three germplasm sources. These lines were grouped as high (RI > 67%) and low restorers (RI < 30%), respectively, using testcrosses with a highly diagnostic CMS tester. Hybrids carrying an exotic restorer gene suffered from a significant grain yield reduction by 4.4% and 9.4% and were 9.3 and 4.8 cm taller in experiments 1 and 2, respectively. Thousand‐kernel weight was reduced, whereas quality traits were only slightly affected. For all traits, significant genetic variance existed among the testcrosses to the presence vs. absence of a given exotic restorer gene. This offers a chance for the breeder to reduce or ultimately overcome the presently observed performance reductions brought about by exotic restorer genes.     

Plant Availability and Uptake of Lead, Zinc, and Cadmium in Soils Contaminated with Anti-corrosion Paint from Pylons in Comparison to Heavy Metal Contaminated Urban Soils

Red lead (Pb₃O₄) has been used extensively in the past as an anti-corrosion paint for the protection of steel constructions. Prominent examples being some of the 200,000 high-voltage pylons in Germany which have been treated with red lead anti-corrosion paints until about 1970. Through weathering and maintenance work, paint compounds and particles are deposited on the soils beneath these constructions. In the present study, six such “pylon soils” were investigated in order to characterize the plant availability and plant uptake of Pb, Cd, and Zn. For comparison, three urban soils with similar levels of heavy metal contamination were included. One phase extractions with 1 M NH₄NO₃, sequential extractions (seven steps), and extractions at different soil pH were used to evaluate the heavy metal binding forms in the soil and availability to plants. Greenhouse experiments were conducted to determine heavy metal uptake by Lolium multiflorum and Lactuca sativa var. crispa in untreated and limed red lead paint contaminated soils. Concentrations of Pb and Zn in the pylon soils were elevated with maximum values of 783 mg Pb kg⁻¹ and 635 Zn mg kg⁻¹ while the soil Cd content was similar to nearby reference soils. The pylon soils were characterized by exceptionally high proportions of NH₄NO₃-extractable Pb reaching up to 17% of total Pb. Even if the relatively low pH of the soils is considered (pH 4.3–4.9), this appears to be a specific feature of the red lead contamination since similarly contaminated urban soils have to be acidified to pH 2.5 to achieve a similarly high Pb extractability. The Pb content in L. multiflorum shoots reached maximum values of 73 mg kg⁻¹ after a cultivation time of 4 weeks in pylon soil. Lime amendment reduced the plant uptake of Pb and Zn significantly by up to 91%. But L. sativa var. crispa cultivated on soils limed to neutral pH still contained critical Pb concentrations (up to 0.6 mg kg⁻¹ fresh weight). Possible mechanisms for the exceptionally high plant availability of soil Pb derived from red lead paint are discussed.