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Mcdonald, Daniel ; Meddi, Franco ; Menchaca-Rocha, Arturo Alejandro ; Meninno, Elisa ; Mercado-Perez, Jorge ; Meres, Michal ; Miake, Yasuo ; Mieskolainen, Matti Mikael ; Mikhaylov, Konstantin ; Milano, Leonardo ; Milosevic, Jovan ; Minervini, Lazzaro Manlio ; Mischke, Andre ; Mishra, Aditya Nath ; Miskowiec, Dariusz Czeslaw ; Mitra, Jubin ; Mitu, Ciprian Mihai ; Mohammadi, Naghmeh ; Mohanty, Bedangadas ; Molnar, Levente ; Montano Zetina, Luis Manuel ; Montes Prado, Esther ; Morando, Maurizio ; Moreira De Godoy, Denise Aparecida ; Moretto, Sandra ; Morreale, Astrid ; Morsch, Andreas ; Muccifora, Valeria ; Mudnic, Eugen ; Mühlheim, Daniel ; Muhuri, Sanjib ; Mukherjee, Maitreyee ; Muller, Hans ; Mulligan, James Declan ; Gameiro Munhoz, Marcelo ; Murray, Sean ; Musa, Luciano ; Musinsky, Jan ; Nandi, Basanta Kumar ; Nania, Rosario ; Nappi, Eugenio ; Naru, Muhammad Umair ; Nattrass, Christine ; Nayak, Kishora ; Nayak, Tapan Kumar ; Nazarenko, Sergey ; Nedosekin, Alexander ; Nellen, Lukas ; Ng, Fabian ; Nicassio, Maria ; Niculescu, Mihai ; Niedziela, Jeremi ; Nielsen, Borge Svane ; Nikolaev, Sergey ; Nikulin, Sergey ; Nikulin, Vladimir ; Noferini, Francesco ; Nomokonov, Petr ; Nooren, Gerardus ; Norman, Jaime ; Nyanin, Alexander ; Nystrand, Joakim Ingemar ; Oeschler, Helmut Oskar ; Oh, Saehanseul ; Oh, Sun Kun ; Ohlson, Alice ; Okatan, Ali ; Okubo, Tsubasa ; Olah, Laszlo ; Oleniacz, Janusz ; Oliveira Da Silva, Antonio Carlos ; Oliver, Michael Henry ; Onderwaater, Jacobus ; Oppedisano, Chiara ; Ortiz Velasquez, Antonio ; Oskarsson, Anders Nils Erik ; Otwinowski, Jacek Tomasz ; Oyama, Ken ; Ozdemir, Mahmut ; Pachmayer, Yvonne C. ; Pagano, Paola ; Paic, Guy ; Pajares Vales, Carlos ; Pal, Susanta Kumar ; Pan, Jinjin ; Pandey, Ashutosh Kumar ; Pant, Divyash ; Papikyan, Vardanush ; Pappalardo, Giuseppe ; Pareek, Pooja ; Park, Woojin ; Parmar, Sonia ; Passfeld, Annika ; Paticchio, Vincenzo ; Patra, Rajendra Nath ; Paul, Biswarup ; Peitzmann, Thomas ; Pereira Da Costa, Hugo Denis Antonio ; Pereira De Oliveira Filho, Elienos ; Peresunko, Dmitry Yurevich ; Perez Lara, Carlos Eugenio ; Peskov, Vladimir ; Pestov, Yury ; Petracek, Vojtech ; Petrov, Viacheslav ; Petrovici, Mihai ; Petta, Catia ; Piano, Stefano ; Pikna, Miroslav ; Pillot, Philippe ; Pinazza, Ombretta ; Pinsky, Lawrence ; Piyarathna, Danthasinghe ; Ploskon, Mateusz Andrzej ; Planinic, Mirko ; Pluta, Jan Marian ; Pochybova, Sona ; Podesta Lerma, Pedro Luis Manuel ; Poghosyan, Martin ; Polishchuk, Boris ; Poljak, Nikola ; Poonsawat, Wanchaloem ; Pop, Amalia ; Porteboeuf, Sarah Julie ; Porter, R Jefferson ; Pospisil, Jan ; Prasad, Sidharth Kumar ; Preghenella, Roberto ; Prino, Francesco ; Pruneau, Claude Andre ; Pshenichnov, Igor ; Puccio, Maximiliano ; Puddu, Giovanna ; Pujahari, Prabhat Ranjan ; Punin, Valery ; Putschke, Jorn Henning ; Qvigstad, Henrik ; Rachevski, Alexandre ; Raha, Sibaji ; Rajput, Sonia ; Rak, Jan ; Rakotozafindrabe, Andry Malala ; Ramello, Luciano ; Raniwala, Rashmi ; Raniwala, Sudhir ; Rasanen, Sami Sakari ; Rascanu, Bogdan Theodor ; Rathee, Deepika ; Read, Kenneth Francis ; Real, Jean-Sebastien ; Redlich, Krzysztof ; Reed, Rosi Jan ; Rehman, Attiq Ur ; Reichelt, Patrick Simon ; Reidt, Felix ; Ren, Xiaowen ; Renfordt, Rainer Arno Ernst ; Reolon, Anna Rita ; Reshetin, Andrey ; Rettig, Felix Vincenz ; Revol, Jean-Pierre ; Reygers, Klaus Johannes ; Riabov, Viktor ; Ricci, Renato Angelo ; Richert, Tuva Ora Herenui ; Richter, Matthias ; Riedler, Petra ; Riegler, Werner ; Riggi, Francesco ; Ristea, Catalin-Lucian ; Rivetti, Angelo ; Rocco, Elena ; Rodriguez Cahuantzi, Mario ; Rodriguez Manso, Alis ; Roeed, Ketil ; Rogochaya, Elena ; Rohr, David ; Roehrich, Dieter ; Romita, Rosa ; Ronchetti, Federico ; Ronflette, Lucile ; Rosnet, Philippe ; Rossi, Andrea ; Roukoutakis, Filimon ; Roy, Ankhi ; Roy, Christelle Sophie ; Roy, Pradip Kumar ; Rubio Montero, Antonio Juan ; Rui, Rinaldo ; Russo, Riccardo ; Ryabinkin, Evgeny ; Ryabov, Yury ; Rybicki, Andrzej ; Sadovskiy, Sergey ; Safarik, Karel ; Sahlmuller, Baldo ; Sahoo, Pragati ; Sahoo, Raghunath ; Sahoo, Sarita ; Sahu, Pradip Kumar ; Saini, Jogender ; Sakai, Shingo ; Saleh, Mohammad Ahmad ; Salgado Lopez, Carlos Alberto ; Salzwedel, Jai Samuel Nielsen ; Sambyal, Sanjeev Singh ; Samsonov, Vladimir ; Sanchez Castro, Xitzel ; Sandor, Ladislav ; Sandoval, Andres ; Sano, Masato ; Santagati, Gianluca ; Sarkar, Debojit ; Scapparone, Eugenio ; Scarlassara, Fernando ; Scharenberg, Rolf Paul ; Schiaua, Claudiu Cornel ; Schicker, Rainer ; Schmidt, Christian Joachim ; Schmidt, Hans Rudolf ; Schuchmann, Simone ; Schukraft, Jürgen ; Schulc, Martin ; Schuster, Tim Robin ; Schutz, Yves Roland ; Schwarz, Kilian Eberhard ; Schweda, Kai ; Scioli, Gilda ; Scomparin, Enrico ; Scott, Rebecca Michelle ; Seeder, Karin Soraya ; Seger, Janet Elizabeth ; Sekiguchi, Yuko ; Selyuzhenkov, Ilya ; Senosi, Kgotlaesele ; Seo, Jeewon ; Serradilla Rodriguez, Eulogio ; Sevcenco, Adrian ; Shabanov, Arseniy ; Shabetai, Alexandre ; Shadura, Oksana ; Shahoyan, Ruben ; Shangaraev, Artem ; Sharma, Ankita ; Sharma, Natasha ; Shigaki, Kenta ; Shtejer Diaz, Katherin ; Sibiryak, Yury ; Siddhanta, Sabyasachi ; Sielewicz, Krzysztof Marek ; Siemiarczuk, Teodor ; Silvermyr, David Olle Rickard ; Silvestre, Catherine Micaela ; Simatovic, Goran ; Simonetti, Giuseppe ; Singaraju, Rama Narayana ; Singh, Ranbir ; Singha, Subhash ; Singhal, Vikas ; Sinha, Bikash ; Sarkar-Sinha, Tinku ; Sitar, Branislav ; Sitta, Mario ; Skaali, Toralf Bernhard ; Skjerdal, Kyrre ; Slupecki, Maciej ; Smirnov, Nikolai ; Snellings, Raimond ; Snellman, Tomas Wilhelm ; Soegaard, Carsten ; Soltz, Ron Ariel ; Song, Jihye ; Song, Myunggeun ; Song, Zixuan ; Soramel, Francesca ; Sorensen, Soren Pontoppidan ; Spacek, Michal ; Spiriti, Eleuterio ; Sputowska, Iwona Anna ; Spyropoulou-Stassinaki, Martha ; Srivastava, Brijesh Kumar ; Stachel, Johanna ; Stan, Ionel ; Stefanek, Grzegorz ; Steinpreis, Matthew Donald ; Stenlund, Evert Anders ; Steyn, Gideon Francois ; Stiller, Johannes Hendrik ; Stocco, Diego ; Strmen, Peter ; Suaide, Alexandre Alarcon do Passo ; Sugitate, Toru ; Suire, Christophe Pierre ; Suleymanov, Mais Kazim Oglu ; Sultanov, Rishat ; Sumbera, Michal ; Symons, Timothy ; Szabo, Alexander ; Szanto De Toledo, Alejandro ; Szarka, Imrich ; Szczepankiewicz, Adam ; Szymanski, Maciej ; Takahashi, Jun ; Tanaka, Naoto ; Tangaro, Marco-Antonio ; Tapia Takaki, Jesus Daniel ; Tarantola Peloni, Attilio ; Tariq, Mohammad ; Tarzila, Madalina-Gabriela ; Tauro, Arturo ; Tejeda Munoz, Guillermo ; Telesca, Adriana ; Terasaki, Kohei ; Terrevoli, Cristina ; Teyssier, Boris ; Thaeder, Jochen Mathias ; Thomas, Deepa ; Tieulent, Raphael Noel ; Timmins, Anthony Robert ; Toia, Alberica ; Trogolo, Stefano ; Trubnikov, Victor ; Trzaska, Wladyslaw Henryk ; Tsuji, Tomoya ; Tumkin, Alexandr ; Turrisi, Rosario ; Tveter, Trine Spedstad ; Ullaland, Kjetil ; Uras, Antonio ; Usai, Gianluca ; Utrobicic, Antonija ; Vajzer, Michal ; Vala, Martin ; Valencia Palomo, Lizardo ; Vallero, Sara ; Van Der Maarel, Jasper ; Van Hoorne, Jacobus Willem ; Van Leeuwen, Marco ; Vanat, Tomas ; Vande Vyvre, Pierre ; Varga, Dezso ; Diozcora Vargas Trevino, Aurora ; Vargyas, Marton ; Varma, Raghava ; Vasileiou, Maria ; Vasiliev, Andrey ; Vauthier, Astrid ; Vechernin, Vladimir ; Veen, Annelies Marianne ; Veldhoen, Misha ; Velure, Arild ; Venaruzzo, Massimo ; Vercellin, Ermanno ; Vergara Limon, Sergio ; Vernet, Renaud ; Verweij, Marta ; Vickovic, Linda ; Viesti, Giuseppe ; Viinikainen, Jussi Samuli ; Vilakazi, Zabulon ; Villalobos Baillie, Orlando ; Vinogradov, Alexander ; Vinogradov, Leonid ; Vinogradov, Yury ; Virgili, Tiziano ; Vislavicius, Vytautas ; Viyogi, Yogendra ; Vodopyanov, Alexander ; Völkl, Martin Andreas ; Voloshin, Kirill ; Voloshin, Sergey ; Volpe, Giacomo ; Von Haller, Barthelemy ; Vorobyev, Ivan ; Vranic, Danilo ; Vrlakova, Janka ; Vulpescu, Bogdan ; Vyushin, Alexey ; Wagner, Boris ; Wagner, Jan ; Wang, Hongkai ; Wang, Mengliang ; Wang, Yifei ; Watanabe, Daisuke ; Weber, Michael ; Weber, Steffen Georg ; Wessels, Johannes Peter ; Westerhoff, Uwe ; Wiechula, Jens ; Wikne, Jon ; Wilde, Martin Rudolf ; Wilk, Grzegorz Andrzej ; Wilkinson, Jeremy John ; Williams, Crispin ; Windelband, Bernd ; Winn, Michael ; Yaldo, Chris G ; Yamaguchi, Yorito ; Yang, Hongyan ; Yang, Ping ; Yano, Satoshi ; Yin, Zhongbao ; Yokoyama, Hiroki ; Yoo, In-Kwon ; Yurchenko, Volodymyr ; Yushmanov, Igor ; Zaborowska, Anna ; Zaccolo, Valentina ; Zaman, Ali ; Zampolli, Chiara ; Correia Zanoli, Henrique Jose ; Zaporozhets, Sergey ; Zarochentsev, Andrey ; Zavada, Petr ; Zavyalov, Nikolay ; Zbroszczyk, Hanna Paulina ; Zgura, Sorin Ion ; Zhalov, Mikhail ; Zhang, Haitao ; Zhang, Xiaoming ; Zhang, Yonghong ; Zhao, Chengxin ; Zhigareva, Natalia ; Zhou, Daicui ; Zhou, You ; Zhou, Zhuo ; Zhu, Hongsheng ; Zhu, Jianhui ; Zhu, Xiangrong ; Zichichi, Antonino ; Zimmermann, Alice ; Zimmermann, Markus Bernhard ; Zinovjev, Gennady ; Zyzak, Maksym
We have has performed the first measurement of the coherent ψ(2S) photo-production cross section in ultra-peripheral Pb-Pb collisions at the LHC. This charmonium excited state is reconstructed via the ψ(2S) →l+l− and ψ(2S) → J/ψπ+π− decays, where the J/ψ decays into two leptons. The analysis is based on an event sample corresponding to an integrated luminosity of about 22 μb−1. The cross section for coherent ψ(2S) production in the rapidity interval −0.9<y<0.9 is dσcohψ(2S)/dy=0.83±0.19(stat+syst) mb. The ψ(2S) to J/ψ coherent cross section ratio is 0.34+0.08−0.07(stat+syst). The obtained results are compared to predictions from theoretical models.
Dick, Rolf van ; Wieland, Volker ; Eckert, Sandra ; Volkmann, Uwe ; Stürmer, Martin ; Seel, Martin ; Andresen, Sabine ; Oswald, Frank ; Wanka, Anna ; Müller, Jürgen ; Gugutzer, Robert
Na+ homeostasis in Acinetobacter baumannii is facilitated via the activity of the Mrp antiporter
(2022)
The human opportunistic pathogen Acinetobacter baumannii is a global threat to healthcare institutions worldwide, since it developed very efficient strategies to evade host defence and to adapt to the different environmental conditions of the host. This work focused on the importance of Na+ homeostasis in A. baumannii with regards to pathobiological aspects. In silico studies revealed a homologue of a multicomponent Na+/H+ antiporter system. Inactivation of the Mrp antiporter through deletion of the first gene (mrpA′) resulted in a mutant that was sensitive to increasing pH values. Furthermore, the strain was highly sensitive to increasing Na+ and Li+ concentrations. Increasing Na+ sensitivity is thought to be responsible for growth impairment in human fluids. Furthermore, deletion of mrpA′ is associated with energetic defects, inhibition of motility and survival under anoxic and dry conditions.
The Rnf complex is a Na+ coupled respiratory enzyme in a fermenting bacterium, Thermotoga maritima
(2020)
rnf genes are widespread in bacteria and biochemical and genetic data are in line with the hypothesis that they encode a membrane-bound enzyme that oxidizes reduced ferredoxin and reduces NAD and vice versa, coupled to ion transport across the cytoplasmic membrane. The Rnf complex is of critical importance in many bacteria for energy conservation but also for reverse electron transport to drive ferredoxin reduction. However, the enzyme has never been purified and thus, ion transport could not be demonstrated yet. Here, we have purified the Rnf complex from the anaerobic, fermenting thermophilic bacterium Thermotoga maritima and show that is a primary Na+ pump. These studies provide the proof that the Rnf complex is indeed an ion (Na+) translocating, respiratory enzyme. Together with a Na+-F1FO ATP synthase it builds a simple, two-limb respiratory chain in T. maritima. The physiological role of electron transport phosphorylation in a fermenting bacterium is discussed.
Butyrate production in the acetogen Eubacterium limosum is dependent on the carbon and energy source
(2021)
Eubacterium limosum KIST612 is one of the few acetogenic bacteria that has the genes encoding for butyrate synthesis from acetyl-CoA, and indeed, E. limosum KIST612 is known to produce butyrate from CO but not from H2 + CO2. Butyrate production from CO was only seen in bioreactors with cell recycling or in batch cultures with addition of acetate. Here, we present detailed study on growth of E. limosum KIST612 on different carbon and energy sources with the goal, to find other substrates that lead to butyrate formation. Batch fermentations in serum bottles revealed that acetate was the major product under all conditions investigated. Butyrate formation from the C1 compounds carbon dioxide and hydrogen, carbon monoxide or formate was not observed. However, growth on glucose led to butyrate formation, but only in the stationary growth phase. A maximum of 4.3 mM butyrate was observed, corresponding to a butyrate:glucose ratio of 0.21:1 and a butyrate:acetate ratio of 0.14:1. Interestingly, growth on the C1 substrate methanol also led to butyrate formation in the stationary growth phase with a butyrate:methanol ratio of 0.17:1 and a butyrate:acetate ratio of 0.33:1. Since methanol can be produced chemically from carbon dioxide, this offers the possibility for a combined chemical-biochemical production of butyrate from H2 + CO2 using this acetogenic biocatalyst. With the advent of genetic methods in acetogens, butanol production from methanol maybe possible as well.
The pyruvate:ferredoxin oxidoreductase of the thermophilic acetogen, Thermoanaerobacter kivui
(2021)
Pyruvate:ferredoxin oxidoreductase (PFOR) is a key enzyme in bacterial anaerobic metabolism. Since a low-potential ferredoxin (Fd2−) is used as electron carrier, PFOR allows for hydrogen evolution during heterotrophic growth as well as pyruvate synthesis during lithoautotrophic growth. The thermophilic acetogenic model bacterium Thermoanaerobacter kivui can use both modes of lifestyle, but the nature of the PFOR in this organism was previously unestablished. Here, we have isolated PFOR to apparent homogeneity from cells grown on glucose. Peptide mass fingerprinting revealed that it is encoded by pfor1. PFOR uses pyruvate as an electron donor and methylene blue (1.8 U·mg−1) and ferredoxin (Fd; 27.2 U·mg−1) as electron acceptors, and the reaction is dependent on thiamine pyrophosphate, pyruvate, coenzyme A, and Fd. The pH and temperature optima were 7.5 and 66 °C, respectively. We detected 13.6 mol of iron·mol of protein−1, consistent with the presence of three predicted [4Fe–4S] clusters. The ability to provide reduced Fd makes PFOR an interesting auxiliary enzyme for enzyme assays. To simplify and speed up the purification procedure, we established a protocol for homologous protein production in T. kivui. Therefore, pfor1 was cloned and expressed in T. kivui and the encoded protein containing a genetically engineered His-tag was purified in only two steps to apparent homogeneity. The homologously produced PFOR1 had the same properties as the enzyme from T. kivui. The enzyme can be used as auxiliary enzyme in enzymatic assays that require reduced Fd as electron donor, such as electron-bifurcating enzymes, to keep a constant level of reduced Fd.
Species of the genus Blautia are typical inhabitants of the human gut and considered as beneficial gut microbes. However, their role in the gut microbiome and their metabolic features are poorly understood. Blautia schinkii was described as an acetogenic bacterium, characterized by a functional Wood–Ljungdahl pathway (WLP) of acetogenesis from H2 + CO2. Here we report that two relatives, Blautia luti and Blautia wexlerae do not grow on H2 + CO2. Inspection of the genome sequence revealed all genes of the WLP except genes encoding a formate dehydrogenase and an electron-bifurcating hydrogenase. Enzyme assays confirmed this prediction. Accordingly, resting cells neither converted H2 + CO2 nor H2 + HCOOH + CO2 to acetate. Carbon monoxide is an intermediate of the WLP and substrate for many acetogens. Blautia luti and B. wexlerae had an active CO dehydrogenase and resting cells performed acetogenesis from HCOOH + CO2 + CO, demonstrating a functional WLP. Bioinformatic analyses revealed that many Blautia strains as well as other gut acetogens lack formate dehydrogenases and hydrogenases. Thus, the use of formate instead of H2 + CO2 as an interspecies hydrogen and electron carrier seems to be more common in the gut microbiome.
Müller, Mareike ; Merz, Felicitas ; Durante, Marco ; Seifert, Volker ; Rieger, Johannes ; Mittelbronn, Michel Guy André ; Harter, Patrick Nikolaus ; Mishustin, Igor N. ; Pshenichnov, Igor A. ; Rödel, Franz ; Stöcker, Horst ; Schopow, Kosta ; Dehghani, Faramarz ; Taucher-Scholz, Gisela ; Bechmann, Ingo
Background: In this interdisciplinary project, the biological effects of heavy ions are compared to those of X-rays using tissue slice culture preparations from rodents and humans. Advantages of this biological model are the conservation of an organotypic environment and the independency from genetic immortalization strategies used to generate cell lines. Its open access allows easy treatment and observation via live-imaging microscopy. Materials and methods: Rat brains and human brain tumor tissue are cut into 300 micro m thick tissue slices. These slices are cultivated using a membrane-based culture system and kept in an incubator at 37°C until treatment. The slices are treated with X-rays at the radiation facility of the University Hospital in Frankfurt at doses of up to 40 Gy. The heavy ion irradiations were performed at the UNILAC facility at GSI with different ions of 11.4 A MeV and fluences ranging from 0.5–10 x 106 particles/cm². Using 3D-confocal microscopy, cell-death and immune cell activation of the irradiated slices are analyzed. Planning of the irradiation experiments is done with simulation programs developed at GSI and FIAS. Results: After receiving a single application of either X-rays or heavy ions, slices were kept in culture for up to 9d post irradiation. DNA damage was visualized using gamma H2AXstaining. Here, a dose-dependent increase and time-dependent decrease could clearly be observed for the X-ray irradiation. Slices irradiated with heavy ions showed less gamma H2AX-positive cells distributed evenly throughout the slice, even though particles were calculated to penetrate only 90–100 micro m into the slice. Conclusions: Single irradiations of brain tissue, even at high doses of 40 Gy, will result neither in tissue damage visible on a macroscopic level nor necrosis. This is in line with the view that the brain is highly radio-resistant. However, DNA damage can be detected very well in tissue slices using gamma H2AX-immuno staining. Thus, slice cultures are an excellent tool to study radiation-induced damage and repair mechanisms in living tissues.
Niu, Minli ; Combs, Stephanie ; Linge, Annett ; Krause, Mechthild ; Baumann, Michael ; Lohaus, Fabian ; Ebert, Nadja ; Tinhofer, Ingeborg ; Budach, Volker ; Müller-von der Grün, Jens ; Rödel, Franz ; Grosu, Anca-Ligia ; Multhoff, Gabriele
Background: Radiochemotherapy (RCT) has been shown to induce changes in immune cell homeostasis which might affect antitumor immune responses. In the present study, we aimed to compare the composition and kinetics of major lymphocyte subsets in the periphery of patients with non-locoregional recurrent (n = 23) and locoregional recurrent (n = 9) squamous cell carcinoma of the head and neck (SCCHN) upon primary RCT. Methods: EDTA-blood of non-locoregional recurrent SCCHN patients was collected before (t0), after application of 20–30 Gy (t1), in the follow-up period 3 (t2) and 6 months (t3) after RCT. In patients with locoregional recurrence blood samples were taken at t0, t1, t2 and at the time of recurrence (t5). EDTA-blood of age-related, healthy volunteers (n = 22) served as a control (Ctrl). Major lymphocyte subpopulations were phenotyped by multiparameter flow cytometry. Results: Patients with non-recurrent SCCHN had significantly lower proportions of CD19+ B cells compared to healthy individuals before start of any therapy (t0) that dropped further until 3 months after RCT (t2), but reached initial levels 6 months after RCT (t3). The proportion of CD3+ T and CD3+/CD4+ T helper cells continuously decreased between t0 and t3, whereas that of CD8+ cytotoxic T cells and CD3+/CD56+ NK-like T cells (NKT) gradually increased in the same period of time in non-recurrent patients. The percentage of CD4+/CD25+/FoxP3+ regulatory T cells (Tregs) decreased directly after RCT, but increased above initial levels in the follow-up period 3 (t2) and 6 (t3) months after RCT. Patients with locoregional recurrence showed similar trends with respect to B, T cells and Tregs between t0 and t5. CD4+ T helper cells remained stably low between t0 and t5 in patients with locoregional recurrence compared to Ctrl. NKT/NK cell subsets (CD56+/CD69+, CD3−/CD56+, CD3−/CD94+, CD3−/NKG2D+, CD3−/NKp30+, CD3−/NKp46+) increased continuously up to 6 months after RCT (t0-t3) in patients without locoregional recurrence, whereas in patients with locoregional recurrence, these subsets remained stably low until time of recurrence (t5). Conclusion: Monitoring the kinetics of lymphocyte subpopulations especially activatory NK cells before and after RCT might provide a clue with respect to the development of an early locoregional recurrence in patients with SCCHN. However, studies with larger patient cohorts are needed. Trial registration: Observational Study on Biomarkers in Head and Neck Cancer (HNprädBio), NCT02059668. Registered on 11 February 2014, https://clinicaltrials.gov/ct2/show/NCT02059668.
Willistein, Max ; Bechtel, Dominique F. ; Müller, Christina S. ; Demmer, Ulrike ; Heimann, Larissa ; Kayastha, Kanwal ; Schünemann, Volker ; Pierik, Antonio J. ; Ullmann, G. Matthias ; Ermler, Ulrich ; Boll, Matthias
Hydride transfers play a crucial role in a multitude of biological redox reactions and are mediated by flavin, deazaflavin or nicotinamide adenine dinucleotide cofactors at standard redox potentials ranging from 0 to –340 mV. 2-Naphthoyl-CoA reductase, a key enzyme of oxygen-independent bacterial naphthalene degradation, uses a low-potential one-electron donor for the two-electron dearomatization of its substrate below the redox limit of known biological hydride transfer processes at E°’ = −493 mV. Here we demonstrate by X-ray structural analyses, QM/MM computational studies, and multiple spectroscopy/activity based titrations that highly cooperative electron transfer (n = 3) from a low-potential one-electron (FAD) to a two-electron (FMN) transferring flavin cofactor is the key to overcome the resonance stabilized aromatic system by hydride transfer in a highly hydrophobic pocket. The results evidence how the protein environment inversely functionalizes two flavins to switch from low-potential one-electron to hydride transfer at the thermodynamic limit of flavin redox chemistry.