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The rise of antimicrobial resistance poses a substantial threat to our health system, and, hence, development of drugs against novel targets is urgently needed. The natural peptide thanatin kills Gram-negative bacteria by targeting proteins of the lipopolysaccharide transport (Lpt) machinery. Using the thanatin scaffold together with phenotypic medicinal chemistry, structural data, and a target-focused approach, we developed antimicrobial peptides with drug-like properties. They exhibit potent activity against Enterobacteriaceae both in vitro and in vivo while eliciting low frequencies of resistance. We show that the peptides bind LptA of both wild-type and thanatin-resistant Escherichia coli and Klebsiella pneumoniae strains with low-nanomolar affinities. Mode of action studies revealed that the antimicrobial activity involves the specific disruption of the Lpt periplasmic protein bridge.

G protein-coupled receptors (GPCRs) are medically important membrane proteins that sample inactive, intermediate, and active conformational states characterized by relatively slow interconversions (∼μs– ms). On a faster timescale (∼ps–ns), the conformational landscape of GPCRs is governed by the rapid dynamics of amino acid side chains. Such dynamics are essential for protein functions such as ligand recognition and allostery. Unfortunately, technical challenges have almost entirely precluded the study of side-chain dynamics for GPCRs. Here, we investigate the rapid side-chain dynamics of a thermostabilized α1B-adrenergic receptor (α1B-AR) as probed by methyl relaxation. We determined order parameters for Ile, Leu, and Val methyl groups in the presence of inverse agonists that bind orthosterically (prazosin, tamsulosin) or allosterically (conopeptide ρ-TIA). Despite the differences in the ligands, the receptor’s overall side-chain dynamics are very similar, including those of the apo form. However, ρ-TIA increases the flexibility of Ile1764x56and possibly of Ile2145x49, adjacent to Pro2155x50of the highly conserved P5x50I3x40F6x44motif crucial for receptor activation, suggesting differences in the mechanisms for orthosteric and allosteric receptor inactivation. Overall, increased Ile side-chain rigidity was found for residues closer to the center of the membrane bilayer, correlating with denser packing and lower protein surface exposure. In contrast to two microbial membrane proteins, in α1B-AR Leu exhibited higher flexibility than Ile side chains on average, correlating with the presence of Leu in less densely packed areas and with higher protein-surface exposure than Ile. Our findings demonstrate the feasibility of studying receptor-wide side-chain dynamics in GPCRs to gain functional insights.

The rapid rise of multi-resistant bacteria poses a significant threat to our health system. The development of drugs against novel targets is urgently needed to replenish the clinical arsenal with effective antibiotics. The naturally occurring peptide thanatin kills Gram-negative bacteria by targeting the periplasmic protein bridge, in particular the lipopolysaccharide transport protein A (LptA). Using the thanatin scaffold together with phenotypic medicinal chemistry and a target-focused approach, we developed potent antimicrobial macrocyclic peptides with drug-like properties that exhibited low frequencies for development of resistance both in vitro and in vivo. Binding affinities to Escherichia coli LptA of the thanatin analogs correlated well with minimal inhibitory concentrations for both parent and thanatin-resistant E. coli strains. Mode of action studies revealed that the antimicrobial activity of the antibiotics involves the specific disruption of the periplasmic Lpt protein bridge. Our studies validate the Lpt protein bridge as a novel target for macrocyclic peptide antibiotics that can avoid current mechanisms of drug resistance. The peptides warrant further preclinical and clinical studies addressing WHO priority 1 carbapenem-resistant Enterobacteriaceae, in particular for Klebsiella pneumoniae lung infections.

Rational targeting of proteins involved in controlling cancer cell behavior with small bioactive compounds can accelerate anti-cancer drug discovery. We report the identification of a new small molecule compound inhibitor of the FGFR adaptor protein FRS2. Pharmacophore-based computational screening combined with functional, biophysical, and structural binding analyses, led to the identification of low-molecular weight ligands that interact with the PTB domain of FRS2. By assessing the compounds’ anti-invasion activity in human FGFR-driven cancer cell models, three chemically distinct bioactive molecules were shortlisted for further analysis. A lead compound was selected that specifically repressed FGFR-driven MAPK activation and matrix invasion and displayed on-target activity in cells. Proteome-wide off-target activity of the primary lead was determined and functional in vivo efficacy in an FGFR driven ovarian cancer model confirmed. We propose inhibition of FRS2 by a small molecular PTB domain ligand as a strategy to repress FGF signaling in FGFR-driven human cancers.

α-adrenergic receptors (αARs) are G protein-coupled receptors that regulate vital functions of the cardiovascular and nervous systems. The therapeutic potential of αARs, however, is largely unexploited and hampered by the scarcity of subtype-selective ligands. Moreover, several aminergic drugs either show off-target binding to αARs or fail to interact with the desired subtype. Here, we report the crystal structure of human α1BAR bound to the inverse agonist (+)-cyclazosin, enabled by the fusion to a DARPin crystallization chaperone. The α1BAR structure allows the identification of two unique secondary binding pockets. By structural comparison of α1BAR with α2ARs, and by constructing α1BAR-α2CAR chimeras, we identify residues 3.29 and 6.55 as key determinants of ligand selectivity. Our findings provide a basis for discovery of α1BAR-selective ligands and may guide the optimization of aminergic drugs to prevent off-target binding to αARs, or to elicit a selective interaction with the desired subtype., This study reports the X-ray structure of the α1B-adrenergic G protein-coupled receptor bound to an inverse agonist, and unveils key determinants of subtype-selective ligand binding that may help the design of aminergic drugs with fewer side-effects.

Heteronuclear NMR in combination with isotope labelling is used to study folding of polypeptides induced by metals in the case of metallothioneins, binding of the peptidic allosteric modulator ?-TIA to the human G-protein coupled ?1b adrenergic receptor, the development of therapeutic drugs that interfere with the biosynthesis of the outer membrane of Gram-negative bacteria, and a system in which protein assembly is induced upon peptide addition. NMR in these cases is used to derive precise structural data and to study the dynamics.

The efficiency of Cu(In,Ga)(S,Se)2 (CIGSSe) solar cell absorbers can be increased by the optimization of the Ga/In and S/Se gradients throughout the absorber. Analyzing such gradients is therefore an important method in tracking the effectiveness of process variations. To measure compositional gradients in CIGSSe, energy dispersive X-ray analysis (EDX) with different acceleration energies performed at both the front surface and the backside of delaminated absorbers was used. This procedure allows for the determination of compositional gradients at locations that are millimeters apart and distributed over the entire sample. The method is therefore representative for a large area and yields information about the lateral homogeneity in the millimeter range. The procedure is helpful if methods such as secondary ion-mass (SIMS), time-of-flight SIMS, or glow-discharge optical emission spectrometry (GDOES) are not available. Results of such EDX measurements are compared with GDOES, and they show good agreement. The procedure can also be used in a targeted manner to detect local changes of the gradients in inhomogeneities or points of interest in the µm range. As an example, a comparison between the compositional gradients in the regular absorber and above the laser cut separating the Mo back contact is shown.

Sample preparation for NMR studies of G protein-coupled receptors faces special requirements: Proteins need to be stable for prolonged measurements at elevated temperatures, they should ideally be uniformly labeled with the stable isotopes

Highly dispersive Cu2ZnSnS4 (CZTS) nanoparticles were successfully synthesized by a simple solvothermal route. A low cost, non-vacuum method was used to deposit CZTS nanoparticle ink on glass substrates by a doctor blade process followed by selenization in a tube furnace to form Cu2ZnSn (S,Se)4 (CZTSSe) layers. Different selenization conditions and particle concentrations were considered in order to improve the crystallinity and surface morphology; the annealing temperature was varied between 400°C and 550°C and the annealing time was varied between 5 min and 20 min in a selenium-nitrogen atmosphere. The influence of annealing conditions on structural, compositional, optical and electrical properties of CZTSSe thin films was studied. An improvement in the structural and surface morphology was observed with increasing of annealing temperature (up to 500°C). An enhancement in the crystallinity and surface morphology were observed for thin films annealed for 10–15 min. Absorption study revealed that the band gap energy of as-deposited CZTS thin film was approximately 1.43 eV, while for CZTSSe thin films it ranged from 1.15 eV to 1.34 eV at different annealing temperatures, and from 1.33 eV to 1.38 eV for different annealing times.

Purpose Treating oral cancer with cold atmospheric plasma (CAP) is an evidence-based-medicine level III concept in ongoing cancer research. There is a discussion concerning the potential risk in medical application of CAP due to the formation of free radicals, which may have an adverse impact. After in recent studies the risk of dramatic effects like genotoxic impact was basically excluded, the focus of this discussion is mainly on acute unwanted clinical effects. Methods A retrospective analysis is including 20 patients suffering from locally advanced head and neck cancer and contaminated ulcerations, who underwent palliative treatment with CAP for decontamination. The focus lies on documented side effects related to CAP. Results There are no, mild or moderate unwanted effects related to the application of CAP, especially never life threatening. Conclusion Understanding the lack of severe side effects in plasma medicine, the role of radical oxygen species (ROS) and radical nitrogen species (RNS) is discussed, proposing a model in which CAP is not a direct effector of antitumor action but rather triggers a singlet oxygen-mediated switch-on effect on the specific target, leading to reactivation of intercellular ROS/RNS-dependent apoptosis signaling in tumor cells. As these processes are strictly restricted to the specific targets and as normal tissue is devoid of the required target, it is neither harmed nor affected.

Purpose Cold atmospheric pressure plasma (CAP) is well known for inactivating microbial pathogens and stimulation of tissue regeneration in chronic wounds. Several authors have reported the effectiveness against cancer in different cell lines and animal models. This is the first report of patients with real clinical benefit following application of CAP, not just visible change of the tumor surface but lasting partial remission. The authors discuss the CAP treatment approach and the efficacy for inoperable head and neck cancer patients. Methods The trial enrolled six patients with locally advanced (pT4) squamous cell carcinoma of the oropharynx suffering from open infected ulcerations. Patients were treated with a jet plasma source (kINPen MED, neoplas tools GmbH, Greifswald, Germany) in cycles of 3 single applications (1 min/cm2 from a distance of 8 mm) within 1 week, each followed by an intermittence of 1 week. Results CAP treatment resulted in a reduction in odor and pain medication requirements, in improvement in social function and a positive emotional affect. Further observance revealed partial remission in two patients for at least nine month. Incisional biopsies at remission demonstrate a moderate amount of apoptotic tumor cells and a desmoplastic reaction of the connective tissue. Conclusion The trial demonstrates the clinical relevance of CAP in cancer treatment. There are three approaches for discussion of tumor remission: (i) the role of myeloid cells, (ii) the ROS/RNS model of cellular impact and (iii) the immunogenic cell death model of cancer treatment, and there is a reflection on non-sustainable tumor response due to adapted tumor microenvironment.

Purpose Beside a proven antimicrobial and inflammatory-modulating spectrum of plasma therapy, recent studies point to impressive molecular and cellular effects against tumor cells. However, underlying mechanisms of anticancer effects and improved wound healing, in particular plasma associated influence on microcirculation, have not been sufficiently clarified yet. To date, there is no convincing method for monitoring therapy with cold atmospheric plasma (CAP) in vivo. Methods After cold plasma treatment with kINPen MED a hyperspectral analysis with the TIVITA™ Tissue System (Diaspective Vision, Pepelow, Germany) was performed in two different patient cases. TIVITA™ Tissue System is an innovative hybrid technology that combines imaging and spectroscopy for assessment and documentation of tissue. Results HSI was able to demonstrate CAP associated effects on microcirculation showing a relevant increase of superficial and deeper cutaneous oxygen saturation, hemoglobin concentration and distribution. Conclusion HSI was able to demonstrate CAP-associated effects on microcirculation showing a relevant increase in superficial and deeper skin oxygen saturation, haemoglobin concentration and distribution. As a clinical diagnostic tool, hyperspectral imaging thus provides further information for a better understanding of microcirculatory CAP effects in cancer therapy and wound healing processes.

The constitution of secondary phases in kesterite Cu2ZnSnSe4 (CZTSe) thin films is still a limiting factor for their application in solar cells. Therefore an enhanced understanding of phase formation processes during the fabrication of CZTSe films is required. In this study we present a temperature and film-depth dependent phase analysis of Zn/Sn/Cu precursors on ZnO substrates selenized at different temperatures. A special sample preparation step using a focused ion beam was applied to prepare shallow angle cross sections for depth-resolved Raman profiling of the thin films. At low selenization temperatures multiphase structures are demonstrated and a first formation of CZTSe besides secondary phases at only 250 °C is detected. At high selenization temperatures an accumulation of ZnSe at the interface of CZTSe and ZnO substrates is observed. Furthermore indications for the formation of a thin SnO2 interface layer were found by X-ray diffraction, secondary electron microscopy and energy dispersive X-ray spectrometry.

FGF2, the ligand of FGF receptors (FGFRs), is expressed in the developing and adult brain. FGF2-FGFR1 signaling causes the induction and maintenance of cancer stem cells through ERK-dependent up-regulation of ZEB1 and Olig2 in glioblastoma. In SHH medulloblastoma, Olig2 triggers tumor initiation from GCPs, maintains quiescent stem-like cells during the disease and contributes to tumor outgrowth at recurrence. We found that FGF2-FGFR signaling causes increased growth and tissue invasion through the FGFR adaptor protein FRS2 in SHH and group-3 medulloblastoma 1. Thus, targeting of FGFR-FRS2 signaling could abrogate brain tumor growth and spread by repressing tumor-promoting functions that are induced by microenvironmental FGF2. Using virtual screening combined with functional validation, we identified protein-protein interaction inhibitors (F2i) that bind FRS2 and abrogate FGFR signaling to the MAP-ERK pathway. Consistent with the requirement of FRS2 for pro-invasive signaling downstream of FGFR1 in medulloblastoma, F2i also efficiently block FGF2-induced migration and invasion in medulloblastoma-derived cells. Selected F2i display excellent binding kinetics with a similar Kd as the natural ligand domain of FGFR and cause steric alterations in the targeted protein domain. On-target activity was confirmed by thermal proteome profiling. Neither in silico screening nor empirical testing revealed significant off-target activity of the compounds. No toxicity of F2i was observed in cell-based models with confirmed functional activity on invasion and MAPK activation. Thus, we identified novel, low molecular weight pharmacological protein-protein interaction inhibitors with an excellent potential to specifically block FGFR functions relevant for brain tumor progression. 1. Santhana Kumar et al., CellReports23, 3798–3812.e8 (2018).

Resonance assignments are challenging for membrane proteins due to the size of the lipid/detergent-protein complex and the presence of line-broadening from conformational exchange. As a consequence, many correlations are missing in the triple-resonance NMR experiments typically used for assignments. Herein, we present an approach in which correlations from these solution-state NMR experiments are supplemented by data from 13C unlabeling, single-amino acid type labeling, 4D NOESY data and proximity of moieties to lipids or water in combination with a structure of the protein. These additional data are used to edit the expected peaklists for the automated assignment protocol FLYA, a module of the program package CYANA. We demonstrate application of the protocol to the 262-residue proton pump from archaeal bacteriorhodopsin (bR) in lipid nanodiscs. The lipid-protein assembly is characterized by an overall correlation time of 44 ns. The protocol yielded assignments for 62% of all backbone (H, N, Cα, Cβ, C′) resonances of bR, corresponding to 74% of all observed backbone spin systems, and 60% of the Ala, Met, Ile (δ1), Leu and Val methyl groups, thus enabling to assign a large fraction of the protein without mutagenesis data. Most missing resonances stem from the extracellular half, likely due intermediate exchange line-broadening. Further analysis revealed that missing information of the amino acid type of the preceding residue is the largest problem, and that 4D NOESY experiments are particularly helpful to compensate for that information loss. Electronic supplementary material The online version of this article (10.1007/s10858-019-00289-7) contains supplementary material, which is available to authorized users.

The metastatic potential of tumors depends in part on their response to growth factors available in the tumor microenvironment. We addressed how micro-environmental cues control tumor cell invasion in pediatric medulloblastoma (MB). We show that bFGF promotes MB tumor cell invasion through FGF receptor (FGFR) in vitro and that pharmacological blockade of FGFR represses brain tissue infiltration in vivo. TGF-ß regulates pro-migratory bFGF function in a context-dependent manner. Under low bFGF, the non-canonical TGF-ß pathway causes ROCK activation and cortical translocation of ERK1/2, which antagonizes FGFR signaling by inactivating FGFR substrate 2 (FRS2), and promotes a contractile, non-motile phenotype. Under high bFGF, negative feedback regulation of FRS2 by bFGF-induced ERK1/2 causes repression of the FGFR pathway. Under these conditions, TGF-ß counters inactivation of FRS2 and restores pro-invasive signaling [1]. These findings pinpoint coincidence detection of bFGF and TGF-ß signaling by FRS2 as a mechanism that controls tumor cell invasion and identified targeting of FRS2 as a rational strategy to abrogate aberrant FGFR signaling independent of kinase inhibition. Using computer-assisted compound discovery, we screened chemical libraries for potential small molecule inhibitors of the interaction between FGFR and FRS2. Using functional testing, we selected potential hits that block invasion comparable to current small molecule FGFR kinase inhibitors and that bind FRS2 in the relevant domain. Thus, our study provides first evidence of rationally discovered, precisely acting small molecule interaction inhibitors with the potential to effectively target pro-invasive signaling in MB and other FGFR-driven cancers. 1. Santhana Kumar K et al. Cell Rep. 2018; 23: 3798–812

Die Mund-Kiefer-Gesichts(MKG)-Chirurgie entwickelt kontinuierlich ihre Gesundheitstechnologie weiter; dazu gehoren auch Lasergerate und Plasmaquellen. Prozessorientiertes Qualitatsmanagement durch Weiterbildung bei Anwendung neuer Gesundheitstechnologien. Zusammenstellung wichtiger Standards und Anforderungen an Schulungsprogramme in der klinischen Laser- und Plasmamedizin. Das universitare Weiterbildungsstudium Diploma in Aesthetic Laser Medicine (D.A.L.M) fuhrt mit einem 1999 vom Bildungsministerium anerkannten strukturierten, multizentrischen, industrieunabhangigen, wissenschaftsorientierten Studienprogramm zum postgradualen schildfahigen Abschluss. Im prozessorientierten Qualitatsmanagement der asthetisch orientierten Lasermedizin und daruber hinaus in der Plasmamedizin ist das D.A.L.M.-Programm ein wichtiges Instrument.

The aim of the study was to learn, whether clinical application of cold atmospheric pressure plasma (CAP) is able to cause (i) visible tumor surface effects and (ii) apoptotic cell kill in squamous cell carcinoma and (iii) whether CAP-induced visible tumor surface response occurs as often as CAP-induced apoptotic cell kill. Twelve patients with advanced head and neck cancer and infected ulcerations received locally CAP followed by palliative treatment. Four of them revealed tumor surface response appearing 2 weeks after intervention. The tumor surface response expressed as a flat area with vascular stimulation (type 1) or a contraction of tumor ulceration rims forming recesses covered with scabs, in each case surrounded by tumor tissue in visible progress (type 2). In parallel, 9 patients with the same kind of cancer received CAP before radical tumor resection. Tissue specimens were analyzed for apoptotic cells. Apoptotic cells were detectable and occurred more frequently in tissue areas previously treated with CAP than in untreated areas. Bringing together both findings and placing side by side the frequency of clinical tumor surface response and the frequency of analytically proven apoptotic cell kill, detection of apoptotic cells is as common as clinical tumor surface response. There was no patient showing signs of an enhanced or stimulated tumor growth under influence of CAP. CAP was made applicable by a plasma jet, kINPen(®) MED (neoplas tools GmbH, Greifswald, Germany).

The objective of this work is the processing and characterization of a dense CuInSe2 solar-cell-absorber-layer based on nanoparticulate precursors. Bimetallic copper-indium- and elemental selenium-nanoparticles were synthesized by wet-chemical processes and then dispersed in organic solvents as nano-inks. These inks were then printed into different layer-stacks on a molybdenum coated float-glass-substrate via doctor-blading. The temperature treatment to transform these layer-stacks into dense CISe thin films was investigated, using a face-to-face technique and mechanically applied pressure or the repetition of coating and annealing. All absorber layers were characterized with SEM, EDX and XRD. Dense, coarse grained CuInSe2 layers with a thickness ≈ 7 µm were formed and the application of mechanical pressure shows potential to reduce thickness and sinter together the nanoparticles to large grains of ca. 3 µm in size. The face-to-face-annealing ensured keeping a stoichiometric ratio of (Cu+In) / Se ≈ 1, and can help reducing the content of oxides, even when the annealing is performed in ambient atmosphere. With a repetition of coating and annealing, dense CISe layers could be produced at low temperature of only 350 °C.

Side effect management of CAP-based medical applications requires detailed knowledge about (1) reactive oxygen and nitrogen species (ROS/RNS) in CAP and in CAP-treated fluids, (2) potential interactions of these ROS/RNS, (3) their target structures in nonmalignant and malignant cells, or in microbes, (4) the site of their action in biological systems and (5) the extent of antioxidative counteraction of living cells towards CAP, (6) the impact of ROS/RNS on inter- and intracellular signaling systems and (7) the borderline between signaling and damaging effects of CAP-derived ROS/RNS. The available literature and further studies should allow to define the dose ranges of CAP that cause a beneficial antitumor or antimicrobial effect without detrimental side effects. The rigid analysis of the very few reports on detrimental or nonselective effects of CAP might allow to define the limits of application, whereas the analysis of specific and beneficial CAP effects should be instrumental for further optimization of medical application of CAP. This chapter also points to specific responses of targeted microbes and tumor cells that amplify the initial CAP-mediated signal. They thus allow the use of non-damaging doses of CAP and establishment of a site-specific strong response in defined target structures.

Based on an increasing understanding of cellular and molecular effects clinical use of cold atmospheric shows a constantly and rapidly expanding application horizon. In particular, the field of antimicrobial efficacy can reference to in vitro and in vitro assured treatment successes, opening up interdisciplinary approaches and potentials. Despite all medical advances, surgical site infections are among the most common nosocomial infections worldwide. While the typical causal pathogen spectrum depends on the type of surgery several endogenous and exogenous risk factors have been identified. The chapter provides an overview of selected prevention measures and illustrates current approaches to integrate cold atmospheric plasma into existing anti SSI prevention and therapy concepts. While there is still a need for further evidence-based research the interim evaluation demonstrates the comprehensive potential of cold atmospheric plasma in the fight against this kind of healthcare-associated nosocomial infection including all medical and economic challenges.

Micro- and nanoparticles were synthesized and stabilized in order to process a copper indium diselenide (CISe) solar cell absorber layer. A microwave assisted solvothermal reaction for In2Se3 particles was performed, finding that smaller particles could be obtained in shorter times and at lower temperatures than with a conventional autoclave based solvothermal synthesis. In a second route In2Se3 and Cu2-xSe nanoparticles have been synthesized via aqueous redox reactions and been Zeta potential-stabilized to acquire a nanoparticulate ink. This ink was drop cast in ambient atmosphere and annealed in N2-atmosphere leading to the formation of CISe at lower temperatures than with state-of-the-art processing routes.

This study is a retrospective review of representing clinical follow-up of 12 patients afflicted with advanced squamous cell carcinoma of the head and neck. Herein, we have used novel physical cold atmospheric pressure plasma (CAP) to decontaminate infected cancer ulcerations and evaluated anti-cancer effects. With use of CAP in this cohort, the data suggests: (1) decreased request for pain medication and (2) reduction of typical fetid odor related to (3) reduction of microbial load. In some cases there is (4) superficial partial remission of tumor and even (5) wound healing of infected ulcerations has been observed following CAP exposure. As a result, CAP treatment appears of benefit for select head and neck cancer patients and future work to optimize CAP in the therapeutic armamentarium advances.

Background Carcinogenesis of head and neck tumor is a multiphase process that leads from the initial transformation of normal cells to a clinically manifest cancer with a latency of month to decades. Oral lichen planus (OLP) is one of the common immune-mediated mucocutaneous disease characterized by chronic inflammatory process, in which an immune response attacks the lining epithelium. Several prospective and retrospective studies reported malignant transformation rates of OLP ranging from 0 to 10%. In this clinical study, we examined the therapeutic potential of Cold Atmospheric Plasma (CAP) for the treatment of OLP as intraoral precancerous lesions. Material & Methods For preclinical in-vitro investigations, tissue samples have been collected from healthy and diseased mucosal regions from 10 patients suffering from OLP. For detection of CAP-related cell death, investigation of DNA fragmentation using the TUNEL-assay has been performed after CAP treatment. Supernatant culture medium of tissue samples were analyzed for 13 different cytokines. Furthermore, white cell populations were stained in CAP-treated tissue samples. Saliva was collected for quantification of inflammatory markers. In addition, five patients suffering from intraoral leukoplakia of the cheek have undergone an in-vivo treatment using the plasma jet kINPen MED. Intraoral treatments have been performed for a period of 1 min/cm2 and with a continuous exhausting of gas and salivary. Results No significant enlargement of apoptotic cells could be detected after plasma exposure within the tissue. At the time this abstract was completed no final data concerning changes in cytokines as well as white cell population were available. The in-vivo treatment of OLP tissue revealed a reduction of pain and local inflammation. Especially the treatment of a therapy-resistant erosive OLP of the cheek could achieve pain reduction and a cure of mucosal ulcers. Conclusion Previous studies were primarily concerned with plasma effects on cancer cells and its value in cancer treatment. This is the second report about plasma treatment of intraoral precancerous lesions. It has been shown that CAP could be useful in the treatment of intraoral mucosa diseases, especially OLP as precancerous lesion. The treatment of precancerous lesions could prevent the development of cancer and obviate the need for cancer treatment. A larger number of cases, investigation of long-term effects, and clinical studies are needed to further qualify plasma for treatment of precancerous lesions. Download : Download high-res image (424KB) Download : Download full-size image Figure shows the right cheek of a multimorbid patient suffering from a therapy-resistant erosive OLP before (left), during (middle), and after (right) CAP treatment. All volunteers gave written and informed consent to participate in the laboratory and clinical study.

Roll-to-roll deposition techniques for the fabrication of chalcopyrite solar cells are of major interest and are a promising alternative to state of the art vacuum processes. However, for roll-to-roll processes the preparation of precursor materials like nanoparticle inks is a crucial point. In this work a study on the preparation technique of copper-indium intermetallic nanoparticles was conducted. The preparation of the nanoparticles is based on the chemical reduction of copper and indium cations with sodium borohydride. Different parameters are discussed regarding their influence on (1) size and shape of the nanoparticles, (2) Cu/In ratio within the synthesised nanoparticles and (3) yield of the synthesis. Results show a strong dependency of the Cu/In ratio of the nanoparticles and the yield of the synthesis on the synthesis parameters. The influence of different parameters like (a) the ratio of metal cations to BH4- anions, (b) the Cu2+/In3+ cation ratio within the precursor solution and (c) the dropping rate of the copper-indium precursor solution are discussed. The Cu/In ratio within the nanoparticles can mainly be controlled by the Cu2+/In3+ cation ratio and the dropping rate of the copper-indium precursor solution. The yield of the synthesis shows saturation behaviour depending on the ratio of metal cations to BH4- anions. Shape and size of the nanoparticles are independent of the varied parameters.

We present a study on the temperature and film depth dependent phase formation in Cu-Zn-Sn-Se thin films. Zn/Sn/Cu precursors were selenized at different temperatures, followed by depth-resolved Raman profiling. A high depth resolution for Raman analysis was achieved by a special sample preparation step using a focused ion beam to prepare shallow angle cross sections (SACS) of the absorber. Multi-phase structures were observed at low selenization temperatures with a first formation of the quaternary Cu 2 ZnSnSe 4 at only 250 °C and the existence of Sn x O y in films annealed at 330 °C. At high selenization temperatures up to 560 °C Cu 2 ZnSnSe 4 was the main phase with some traces of ZnSe and a MoSe 2 interface layer at the back contact. Furthermore, compositional gradients were investigated by scanning electron microscopy and energy dispersive X-ray spectroscopy measurements on sample cross sections. The combination of these results allows for observation of both elemental composition and phase composition of the films and their dynamics during the annealing procedure.

Cold atmospheric pressure plasma (CAP) is known to very effectively inactivate multi-resistant strains of microorganisms. Whether or not application of CAP also inactivates cancer cells is a matter of intense clinical interest. There is a need for prospective, randomized, blindly evaluated clinical trials. This paper outlines the key points of such a study program.

Prostate cancer is among the most common cancers. Although it has a relatively good prognosis, 15 to 30% of men with prostate cancer experience a relapse after radical prostatectomy. Identifying patients with an aggressive tumor will therefore help to improve prostate cancer management. DNA methylation of PITX2 has been established in several studies as a prognostic biomarker for breast and prostate cancer. These case control studies were conducted using research assay components; to facilitate its use in a diagnostic setting, the PITX2 biomarker was transferred to a validated diagnostic platform, the Affymetrix GeneChip System. A customized microarray (Epichip PITX2) was designed using features in high redundancy to ensure a robust determination of the methylation state of the PITX2 promoter. The developed method allowed for accurate assessment of prognosis in prostate cancer patients who underwent radical prostatectomy. Determination of PITX2 methylation in formalin-fixed and paraffin-embedded tissue samples from a cohort of 157 prostatectomy patients resulted in an excellent level of concordance of the clinical classification, as well as the measured output between the research assay and the Epichip PITX2. These analytical performance results describe the Epichip PITX2 as a robust and reliable diagnostic tool for assessing the methylation status of PITX2, enabling an improved outcome prediction in cancer patients following radical prostatectomy.

The Judges of the Special Tribunal for Lebanon (STL) recently adopted the Rules of Procedure and Evidence to guide the work of the court in bringing to justice those responsible for the attack of 14 February 2005 that resulted in the death of then-Lebanese Prime Minister Rafiq Hariri (‘the Hariri Attack’) as well as related attacks. These provisions draw heavily on analogous instruments of the International Criminal Court and the International Criminal Tribunals for the former Yugoslavia and Rwanda. However, they also contain a number of innovations, including the enhanced role of the Pre-Trial Judge, the establishment of an independent and empowered Defence Office and the possibility of trials in absentia. The review carried out in this article is not a comprehensive analysis of every provision of the Rules of Procedure and Evidence. Instead, the purpose is to describe the key features of this instrument and, in doing so, to highlight points of interest, intersection and divergence in comparison with the analogous instruments of other International Criminal Tribunals. As such, the exegesis is intended to provide an overview of the procedural framework of the STL that will be of use to scholars and practitioners alike.

Background: The presence of aberrantly methylated SEPT9 DNA in plasma is highly correlated with the occurrence of colorectal cancer. We report the development of a new SEPT9 biomarker assay and its validation in case–control studies. The development of such a minimally invasive blood-based test may help to reduce the current gap in screening coverage. Methods: A new SEPT9 DNA methylation assay was developed for plasma. The assay comprised plasma DNA extraction, bisulfite conversion of DNA, purification of bisulfite-converted DNA, quantification of converted DNA by real-time PCR, and measurement of SEPT9 methylation by real-time PCR. Performance of the SEPT9 assay was established in a study of 97 cases with verified colorectal cancer and 172 healthy controls as verified by colonoscopy. Performance based on predetermined algorithms was validated in an independent blinded study with 90 cases and 155 controls. Results: The SEPT9 assay workflow yielded 1.9 μg/L (CI 1.3–3.0) circulating plasma DNA following bisulfite conversion, a recovery of 45%–50% of genomic DNA, similar to yields in previous studies. The SEPT9 assay successfully identified 72% of cancers at a specificity of 93% in the training study and 68% of cancers at a specificity of 89% in the testing study. Conclusions: Circulating methylated SEPT9 DNA, as measured in the new mSEPT9 assay, is a valuable biomarker for minimally invasive detection of colorectal cancer. The new assay is amenable to automation and standardized use in the clinical laboratory.

Combining the concept of magnetic drug targeting and photodynamic therapy is a promising approach for the treatment of cancer. A high selectivity as well as significant fewer side effects can be achieved by this method, since the therapeutic treatment only takes place in the area where accumulation of the particles by an external electromagnet and radiation by a laser system overlap. In this article, a novel hypericin-bearing drug delivery system has been developed by synthesis of superparamagnetic iron oxide nanoparticles (SPIONs) with a hypericin-linked functionalized dextran coating. For that, sterically stabilized dextran-coated SPIONs were produced by coprecipitation and crosslinking with epichlorohydrin to enhance stability. Carboxymethylation of the dextran shell provided a functionalized platform for linking hypericin via glutaraldehyde. Particle sizes obtained by dynamic light scattering were in a range of 55–85 nm, whereas investigation of single magnetite or maghemite particle diameter was performed by transmission electron microscopy and X-ray diffraction and resulted in approximately 4.5–5.0 nm. Surface chemistry of those particles was evaluated by Fourier transform infrared spectroscopy and ζ potential measurements, indicating successful functionalization and dispersal stabilization due to a mixture of steric and electrostatic repulsion. Flow cytometry revealed no toxicity of pure nanoparticles as well as hypericin without exposure to light on Jurkat T-cells, whereas the combination of hypericin, alone or loaded on particles, with light-induced cell death in a concentration and exposure time-dependent manner due to the generation of reactive oxygen species. In conclusion, the combination of SPIONs’ targeting abilities with hypericin’s phototoxic properties represents a promising approach for merging magnetic drug targeting with photodynamic therapy for the treatment of cancer.

A sequence of two chemoselective reductions enables the stereoselective synthesis of phosphorylated azasugars starting from products of dihydroxyacetonephosphate-dependent aldolases.