Evaluation of Concomitant Systemic Treatment in Older Adults With Head and Neck Squamous Cell Carcinoma Undergoing Definitive Radiotherapy

IMPORTANCE The number of older adults with head and neck squamous cell carcinoma (HNSCC) is increasing, and these patients are underrepresented in clinical trials. It is unclear whether the addition of chemotherapy or cetuximab to radiotherapy is associated with improved survival in older adults with HNSCC. OBJECTIVE To examine whether the addition of chemotherapy or cetuximab to definitive radiotherapy is associated with improved survival in patients with locoregionally advanced (LA) HNSCC. DESIGN, SETTING, AND PARTICIPANTS The Special Care Patterns for Elderly HNSCC Patients Undergoing Radiotherapy (SENIOR) study is an international, multicenter cohort study including older adults (≥65 years) with LA-HNSCCs of the oral cavity, oropharynx/hypopharynx, or larynx treated with definitive radiotherapy, either alone or with concomitant systemic treatment, between January 2005 and December 2019 at 12 academic centers in the US and Europe. Data analysis was conducted from June 4 to August 10, 2022. INTERVENTIONS All patients underwent definitive radiotherapy alone or with concomitant systemic treatment. MAIN OUTCOMES AND MEASURES The primary outcome was overall survival. Secondary outcomes included progression-free survival and locoregional failure rate. RESULTS Among the 1044 patients (734 men [70.3%]; median [IQR] age, 73 [69-78] years) included in this study, 234 patients (22.4%) were treated with radiotherapy alone and 810 patients (77.6%) received concomitant systemic treatment with chemotherapy (677 [64.8%]) or cetuximab (133 [12.7%]). Using inverse probability weighting to attribute for selection bias, chemoradiation was associated with longer overall survival than radiotherapy alone (hazard ratio [HR], 0.61; 95% CI, 0.48-0.77; P < .001), whereas cetuximab-based bioradiotherapy was not (HR, 0.94; 95% CI, 0.70-1.27; P = .70). Progression-free survival was also longer after the addition of chemotherapy (HR, 0.65; 95% CI, 0.52-0.81; P < .001), while the locoregional failure rate was not significantly different (subhazard ratio, 0.62; 95% CI, 0.30-1.26; P = .19). The survival benefit of the chemoradiation group was present in patients up to age 80 years (65-69 years: HR, 0.52; 95% CI, 0.33-0.82; 70-79 years: HR, 0.60; 95% CI, 0.43-0.85), but was absent in patients aged 80 years or older (HR, 0.89; 95% CI, 0.56-1.41). CONCLUSIONS AND RELEVANCE In this cohort study of older adults with LA- HNSCC, chemoradiation, but not cetuximab-based bioradiotherapy, was associated with longer survival compared with radiotherapy alone

Effekte von Landwirtschaft auf gelösten organischen Kohlenstoff und Stickstoff in Fließgewässern

Agricultural land use covers up to 40% of the terrestrial surface. Management practices associated with agricultural land use often alter the soil organic matter status and increase nutrient loads in soils and adjacent aquatic systems. In freshwaters dissolved organic matter (DOM) is the major form of organic matter and plays a key role in various ecological and biogeochemical processes. Thereby dissolved organic carbon (DOC) and nitrogen (DON) constitute main nutrient and energy sources to heterotrophic and autotrophic freshwater biota. As a consequence therof DOM, and in particular DON can contribute to eutrophication and support the growth of harmful phytoplankton. The identification of DOM and DON sources and composition as well as the determination of DOM degradability is therefore crucial e.g., for decision making in water conservation and resources management. In this thesis I aimed to fill the knowledge gaps on the effects of agriculture on DOM export to streams and on DOM processing within streams, focusing in particular on dissolved organic nitrogen. In order to investigate DOM and DON export, two field studies, one at local and one at global scale, were conducted as parts of this thesis. At local scale, DOC and DON amount and composition was monitored in 12 agricultural and forest headwater streams, situated in the Northeastern German Lowlands over a one year period. On global scale, land use effects were evaluated for 75 agricultural and 45 reference streams in 5 different climate zones, including areas with intensive and extensive farming. Thereby, sampling was performed during two main seasons to assess possible temporal variations in DOC and DON amount and composition. Furthermore, in order to enhance the understanding how DOM is further processed, biodegradability of DOM from pristine and agricultural catchments was investigated and the combined effects of DOM composition and inorganic nutrient concentration were studied in a batch experiment with benthic stream biolfilm bacteria. In all field and laboratory studies within this thesis, SEC and optical measurement were applied in parallel for the characterization and quantification of DOM. No information were available on the vulnerability of DOM samples for later SEC analysis to storage effects. Therefore, in this thesis a laboratory experiment provided information on the effects of cold storage and freezing on DOC and DON concentration and size fractions determined by SEC analysis, and therefore contribute to fill these knowledge gaps. Overall, this thesis revelealed that agricultural DOM was more microbially processed and had a much lower C:N ratio, and higher contributions of non-humic high molecular weight DOM than its forest counterpart. This was observed on local, as well as on global scale and presumably a consequence of accelarated organic matter processing in agricultural soils resulting from management processess, in particular fertilization. Strongly elevated DON loads from agricultural catchments indicate that agricultural soils can constitute important sources of DON and nitrogen-rich terrestrial DOM in streams. Even though the low C:N ratio of agricultural DOM was indicative of a lower content of refractory DOM sources in agricultural DOM, degradation of DOM was not evident during 81 days of laboratory experiment in this thesis. The absence of biodegradation of DOM by a benthic stream biofilm was likely not a result of the refractory character of DOM, but presumably can be attributed to the independence of the biofilm bacteria from external sources. However, from the results of the bioassay in this thesis no general conclusions on the biodegradability of DOM in freshwaters and its dependence on either DOM composition, or nutrient availability can be drawn. On the other hand, the findings indicated potentially different responses of planktonic versus benthic biofilm bacteria on DOM. Finally the findings of this thesis call for the integration of DON losses from agricultural soils to considerations in catchment nutrient management, as well as in terrestrial, but also aquatic N budgets. To completely unravel the fate and role of terrestrial N-rich terrestrial DOM in freshwaters, further research should also focus on the availability of this DOM to stream biofilm and planktonic organisms.Landwirtschaftlich genutzte Fläche bedeckt bis zu 40% der globalen Landfläche. Die Bewirtschaftung dieser Flächen geht oft mit Veränderungen des organischen Materials, sowie mit Erhöhung der Nährstofffrachten in Böden, und weiterführend auch in Gewässern einher. In Süßwasser spielt gelöstes organisches Material (DOM) eine Schlüsselrolle in vielen ökologischen und biogeochemischen Prozessen. Dabei stellen gelöster organischer Kohlenstoff (DOC) und Stickstoff (DON) eine Hauptquelle für die Versorgung von heterotrophen und autotrophen Süßwasserorganismen mit Nährstoffen und Energie dar. Eine Folge davon ist, dass im speziellen DON stark zu Eutrophierung und dem vermehrten Wachstum von schädlichem Phytoplankton führen kann. Die Quellen und Zusammensetzung von DOM und DON zu identifizieren, ist daher von maßgeblicher Bedeutung, zum Beispiel zur Enscheidungsfindung im Gewässerschutz und der Wasserwirtschaft. Mein Ziel war es, in der vorliegenden Arbeit bestehende Wissenslücken bezüglich des Einflusses von Landwirtschaft auf den Austrag und die Umsetzung von DOM und im speziellen DON, in Fließgewässern zu füllen. Der Austrag von DOM und DON wurde dabei im Rahmen zweier Feldstudien auf lokaler und globaler Ebene untersucht. Dazu wurden auf lokaler Ebene monatlich, für die Dauer eines Jahres, Bäche mit landwirtschaftlicher Nutzung oder Wald im Einzugsgebiet im Nordosten Deutschlands beprobt. Auf globaler Ebene wurden Landnutzungseffekte in Bächen mit landwirtschaftlicher Nutzung und Referenz-Bäche mit naturnaher Nutzung im Einzugsgebiet untersucht. Die auf globaler Ebene untersuchten Einzugsgebiete umfassten Gebiete mit intensiver, wie auch extensiver Landwirtschaft und waren über 5 Klimazonen verteilt. Mit dem Ziel Erkenntnisse über die Umsetzung von DOM und DON aus unterschiedlichen Landnutzungen zu erlangen wurde ein Abbauversuch mit DOM aus landwirtschaftlich genutzten und Waldeinzugsgebieten und unterschiedlich hohen Konzentrationen durchgeführt. In allen Untersuchungen innerhalb dieser Studie wurde zur Bestimmung der DOC und DON Konzentration und DOM Zusammensetzung Größenauschlusschromatographie (SEC) parallel mit der Analyse von Spektralen Eigenschaften des DOM durchgeführt. Die Ergebnisse eines Laborexperimentes zum Effekt von Probenlagerung auf DOC und DON Zusammensetzung, sowie Konzentration schließen Wissenslücken bezüglich der Effekte von Probenlagerung auf die Größenfraktionierung mittels SEC. Insgesamt demonstrierte diese Arbeit deutliche Unterschiede in der DOM Zusammensetzung und dem DON Austrag zwischen Landnutzungen. Dabei konnte herausgestellt werden, dass landwirtschaftliches DOM mikrobiell stärker umgesetzt und durch ein deutlich niedrigeres C:N Verhältnis sowie höhere Anteile an nicht-huminstoffartigen, hochmolekularen Substanzen gekennzeichnet war. Diese Unterschiede fanden sich auf lokaler und globaler Ebene wieder und sind wahrscheinlich das Resultat von verstärkter Umsetzung organischen Materials in landwirtschaftlich genutzten Böden durch Düngung. Die hohen DON und DOM Frachten zeigen, dass Böden eine bedeutende Quelle für Stickstoff-reiches organisches Material in Gewässern in landwirtschaftlichen Einzugsgebieten darstellen. Obwohl ein niedriges C:N Verhältnis auf eine höhere Verfügbarkeit hindeutete, lieferte ein Abbauversuch mit benthischen Biofilmbakterien keinerlei Hinweis auf den Abbau von landwirtschaftlichem DOM, während es Hinweise auf den Abbau durch pelagische Bakterien gab. Die Abwesenheit von Abbauprozesses war dabei weniger ein Resultat der DOM Zusammensetzung, sondern vermutlich eher ein Resultat der Unabhängigkeit von Biofilmbakterien von externen Nährstoffen. Obwohl auf der Grundlage dieser Erkenntnisse keine generellen Schlussfolgerungen bezüglich der Abbaubarkeit von DOM gezogen werden können, deuten die Ergebnisse auf unterschiedliche Reaktivität bezüglich DOM von Biofilmbakterien gegenüber planktonischen Bakterien hin. Abschließend verdeutlicht diese Arbeit, dass der erhöhe DON Austrag aus landwirtschaftlichen Böden in terrestrische, aber auch aquatisch Stickstoffbilanzen, sowie bei der Entscheidungsfindung im Gewässerschutz und Wasserwirtschaft integriert werden sollte. Um die Rolle von N-reichem terrestrischen DOM aus landwirtschaftlichen Gebieten in Gewässern abschließend zu klären, sollten der Fokus künftiger Studien auf der Verfügbarkeit dieses DOM für planktonische und Biofilm-Organismen liegen

Storage effects on quantity and composition of dissolved organic carbon and nitrogen of lake water, leaf leachate and peat soil water

This study aimed to evaluate the effects of freezing and cold storage at 4 °C on bulk dissolved organic carbon (DOC) and nitrogen (DON) concentration and SEC fractions determined with size exclusion chromatography (SEC), as well as on spectral properties of dissolved organic matter (DOM) analyzed with fluorescence spectroscopy. In order to account for differences in DOM composition and source we analyzed storage effects for three different sample types, including a lake water sample representing freshwater DOM, a leaf litter leachate of Phragmites australis representing a terrestrial, ‘fresh’ DOM source and peatland porewater samples. According to our findings one week of cold storage can bias DOC and DON determination. Overall, the determination of DOC and DON concentration with SEC analysis for all three sample types were little susceptible to alterations due to freezing. The findings derived for the sampling locations investigated here may not apply for other sampling locations and/or sample types. However, DOC size fractions and DON concentration of formerly frozen samples should be interpreted with caution when sample concentrations are high. Alteration of some optical properties (HIX and SUVA254) due to freezing were evident, and therefore we recommend immediate analysis of samples for spectral analysis

Global effects of agriculture on fluvial dissolved organic matter

Agricultural land covers approximately 40% of Earth's land surface and affects hydromorphological, biogeochemical and ecological characteristics of fluvial networks. In the northern temperate region, agriculture also strongly affects the amount and molecular composition of dissolved organic matter (DOM), which constitutes the main vector of carbon transport from soils to fluvial networks and to the sea and is involved in a large variety of biogeochemical processes. Here, we provide first evidence about the wider occurrence of agricultural impacts on the concentration and composition of fluvial DOM across climate zones of the northern and southern hemispheres. Both extensive and intensive farming altered fluvial DOM towards a more microbial and less plant-derived composition. Moreover, intensive farming significantly increased dissolved organic nitrogen (DON) concentrations. The DOM composition change and DON concentration increase differed among climate zones and could be related to the intensity of current and historical nitrogen fertilizer use. As a result of agriculture intensification, increased DON concentrations and a more microbial-like DOM composition likely will enhance the reactivity of catchment DOM emissions, thereby fuelling the biogeochemical processing in fluvial networks and resulting in higher ecosystem productivity and CO2 outgassing

Comparison of Organic Matter Composition in Agricultural versus Forest Affected Headwaters with Special Emphasis on Organic Nitrogen

Agricultural management practices promote organic matter (OM) turnover and thus alter both the processing of dissolved organic matter (DOM) in soils and presumably also the export of DOM to headwater streams, which intimately connect the terrestrial with the aquatic environment. Size-exclusion chromatography, in combination with absorbance and emission matrix fluorometry, was applied to assess how agricultural land use alters the amount and composition of DOM, as well as dissolved organic nitrogen (DON) forms in headwater streams, including temporal variations, in a temperate region of NE Germany. By comparing six agriculturally and six forest-impacted headwater streams, we demonstrated that agriculture promotes increased DOC and DON concentrations, entailing an even more pronounced effect on DON. The major part of DOC and DON in agricultural and forest reference streams is exported in the form of humic-like material with high molecular weight, which indicates terrestrial, i.e., allochthonous sources. As an obvious difference in agricultural streams, the contribution of DOC and particularly DON occurring in the form of nonhumic high-molecular-weight, presumably proteinous material is clearly elevated. Altogether, DOM in agricultural headwaters is mainly complex-soil-derived and aromatic material with a low C:N ratio, which is more microbial processed than its counterpart from forest reference catchments. Our results emphasize the importance of agricultural land use on DOM loss from soils and identify agricultural soils as important DOC and particularly DON sources to headwater streams

Effect of A2M* on expression of CD44 and CD29 in different tumour cell lines.

<p><b>(a)</b> 1321N1 and A549 cells were cultured in the absence and presence of increasing concentrations of A2M, lysed and immunoblotted for CD44 and CD29. <b>(b)</b> A549 cells were stimulated by RAP and treated as shown in (a). <b>(c)</b> A549 cells were treated with A2M* and expression of total and cell surface CD44 was analysed by flow cytometry (n = 3), error bars mean ± sem, t-test (*P < 0.05, **P < 0.01). <b>(d)</b> A549 cell were treated with 100 nM A2M* for 12h, fixed by paraformaldehyde and immunestained for CD44 and counterstained with DAPI. <b>(e)</b> Immunoprecipitation of CD44 in A2M*-stimulated and non-stimulated A549 and MDA-MB-231 cells. Cells were treated with 30 nM A2M* for 24h, lysates were precipitated with different antibodies (anti-LRP1, anti-A2M*, anti-GLO1) and Western blotted for CD44 protein. <b>(f)</b> Ligand blot of A2M* for CD44 binding. A2M* was blotted to membranes which were incubated with cell lysate of A549 afore stimulated by 30 nM A2M* for 8h. Binding of CD44 to immobilized A2M* was detected by anti-CD44/HRP-anti-mouse-Ig. a) Immobilized A2M* detected by anti-A2M*/HRP-anti-mouse-Ig and b) Detection of binding of CD44 to immobilized A2M*. <b>(g)</b> Ligand blot demonstrating binding of CD44 to LRP1-associated RAP. Membrane-blotted LRP1 was incubated with A549 cell lysate and bound CD44 was detected by respective antibodies. <b>(h)</b> Recombinant RAP was electro-blotted, followed by incubation with A549 cell lysate. Detection of CD44 bound to immobilized RAP was accomplished by anti-CD44/HRP-anti-mouse-Ig. Immobilized RAP was detected by anti-RAP antibody (blotting control).</p