Effekte einer Adaptiven Servo-Ventilation auf Herzrhythmusstörungen bei Patienten mit chronischer Herzinsuffizienz und schlafbezogenen Atmungsstörungen – Subanalyse einer randomisierten Studie

Patienten mit chronischer Herzinsuffizienz (HI) und schlafbezogenen Atmungsstörungen (SBAS) leiden häufig unter nächtlich auftretenden kardialen Arrhythmien. SBAS können effektiv mit einer Adaptiven Servo-Ventilation (ASV) behandelt werden. In dieser Studie wurde daher die Hypothese überprüft, ob eine ASV Therapie bei Patienten mit HI und SBAS die Häufigkeit nächtlicher kardialer Arrhythmien reduziert. Hierzu wurden zwanzig Patienten mit stabiler HI und SBAS randomisiert und entweder einer ASV-Gruppe oder einer Kontrollgruppe mit alleiniger optimaler medizinischer Versorgung zugeteilt. Zu Baseline und nach 12 Wochen wurde jeweils eine Polysomnographie (PSG) mit zentraler verblindeter Auswertung durchgeführt. Die nächtlichen Elektrokardiogramme (EKG) der PSG wurden manuell und mit Unterstützung einer Langzeit-EKG Software ausgewertet. Für die ASV-Gruppe nahmen Ventrikuläre Extrasystolen (VES) pro Stunde Aufnahmezeit im Gegensatz zur Kontrollgruppe ab. Eine ASV Beatmungstherapie reduziert im Vergleich mit der Kontrollgruppe die Anzahl ventrikulärer Couplets sowie nicht-anhaltender Ventrikulärer Tachykardien. Die Veränderungen waren jeweils nicht statistisch signifikant

Structural Basis of Enzymatic Activity for the Ferulic Acid Decarboxylase (FADase) from Enterobacter sp. Px6-4

Microbial ferulic acid decarboxylase (FADase) catalyzes the transformation of ferulic acid to 4-hydroxy-3-methoxystyrene (4-vinylguaiacol) via non-oxidative decarboxylation. Here we report the crystal structures of the Enterobacter sp. Px6-4 FADase and the enzyme in complex with substrate analogues. Our analyses revealed that FADase possessed a half-opened bottom β-barrel with the catalytic pocket located between the middle of the core β-barrel and the helical bottom. Its structure shared a high degree of similarity with members of the phenolic acid decarboxylase (PAD) superfamily. Structural analysis revealed that FADase catalyzed reactions by an “open-closed” mechanism involving a pocket of 8×8×15 Å dimension on the surface of the enzyme. The active pocket could directly contact the solvent and allow the substrate to enter when induced by substrate analogues. Site-directed mutagenesis showed that the E134A mutation decreased the enzyme activity by more than 60%, and Y21A and Y27A mutations abolished the enzyme activity completely. The combined structural and mutagenesis results suggest that during decarboxylation of ferulic acid by FADase, Trp25 and Tyr27 are required for the entering and proper orientation of the substrate while Glu134 and Asn23 participate in proton transfer

Vanillin derived a carbonate dialdehyde and a carbonate diol : novel platform monomers for sustainable polymers synthesis

Vanillin has been regarded as one of the important biomass-based platform chemicals for aromatic polymers synthesis. Herein, novel symmetric bis(4-formyl-2-methoxyphenyl)carbonate (BFMC) and bis(4-(hydroxymethyl)-2-methoxyphenyl)carbonate (BHMC) polymeric monomers have been synthesized in high yields using vanillin as a raw chemical, which have been submitted for polymer synthesis via well-established polymeric strategies. A new class of poly(carbonate ester)s oligomers with amide moieties in their side chain can be prepared by using the BFMC as one of monomers via the Passerini three compound reaction (3CR). A new class of poly(carbonate ester)s oligomers and poly(carbonate urethane)s can be prepared via reactions between BHMC with dicarboxylic acid chlorides and diisocyanates, respectively. Their structure have been confirmed by 1H NMR, 13C NMR and FTIR, and the gel permeation chromatograph (GPC) analysis shows that the Mn of poly(carbonate ester)s oligomers ranges from 3100 to 7900 with PDI between 1.31 and 1.65, and the Mn of poly(carbonate urethane)s ranges from 16 400 to 24 400 with PDI ranging from 1.36 to 2.17. The DSC analysis shows that the poly(carbonate ester)s oligomers have relative low Tg ranging from 37.4 to 74.1 °C, and the poly(carbonate urethane)s have Tg ranging from 97.3 to 138.3 °C, mainly correlating to the structure of dicarboxylic acid chlorides and diisocyanates used

Applications of lignin in the agri-food industry

Of late, valorization of agri-food industrial by-products and their sustainable utilization is gaining much contemplation world-over. Globally, 'Zero Waste Concept' is promoted with main emphasis laid towards generation of minimal wastes and maximal utilization of plantbased agri-food raw materials. One of the wastes/by-products in the agri-food industry are the lignin, which occurs as lignocellulosic biomass. This biomass is deliberated to be an environmental pollutant as they offer resistance to natural biodegradation. Safe disposal of this biomass is often considered a major challenge, especially in low-income countries. Hence, the application of modern technologies to effectively reduce these types of wastes and maximize their potential use/applications is vital in the present day scenario. Nevertheless, in some of the high-income countries, attempts have been made to efficiently utilize lignin as a source of fuel, as a raw material in the paper industry, as a filler material in biopolymer based packaging and for producing bioethanol. However, as of today, agri-food industrial applications remains significantly underexplored. Chemically, lignin is heterogeneous, bio-polymeric, polyphenolic compound, which is present naturally in plants, providing mechanical strength and rigidity. Reports are available wherein purified lignin is established to possess therapeutic values; and are rich in antioxidant, anti-microbial, anti-carcinogenic, antidiabetic properties, etc. This chapter is divided into four sub-categories focusing on various technological aspects related to isolation and characterization of lignin; established uses of lignin; proved bioactivities and therapeutic potentials of lignin, and finally on identifying the existing research gaps followed by future recommendations for potential use from agri-food industrial wastes.Theme of this chapter is based on our ongoing project- Valortech, which has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No 810630

Complete Genome Sequence of the N2-Fixing Broad Host Range Endophyte Klebsiella pneumoniae 342 and Virulence Predictions Verified in Mice

We report here the sequencing and analysis of the genome of the nitrogen-fixing endophyte, Klebsiella pneumoniae 342. Although K. pneumoniae 342 is a member of the enteric bacteria, it serves as a model for studies of endophytic, plant-bacterial associations due to its efficient colonization of plant tissues (including maize and wheat, two of the most important crops in the world), while maintaining a mutualistic relationship that encompasses supplying organic nitrogen to the host plant. Genomic analysis examined K. pneumoniae 342 for the presence of previously identified genes from other bacteria involved in colonization of, or growth in, plants. From this set, approximately one-third were identified in K. pneumoniae 342, suggesting additional factors most likely contribute to its endophytic lifestyle. Comparative genome analyses were used to provide new insights into this question. Results included the identification of metabolic pathways and other features devoted to processing plant-derived cellulosic and aromatic compounds, and a robust complement of transport genes (15.4%), one of the highest percentages in bacterial genomes sequenced. Although virulence and antibiotic resistance genes were predicted, experiments conducted using mouse models showed pathogenicity to be attenuated in this strain. Comparative genomic analyses with the presumed human pathogen K. pneumoniae MGH78578 revealed that MGH78578 apparently cannot fix nitrogen, and the distribution of genes essential to surface attachment, secretion, transport, and regulation and signaling varied between each genome, which may indicate critical divergences between the strains that influence their preferred host ranges and lifestyles (endophytic plant associations for K. pneumoniae 342 and presumably human pathogenesis for MGH78578). Little genome information is available concerning endophytic bacteria. The K. pneumoniae 342 genome will drive new research into this less-understood, but important category of bacterial-plant host relationships, which could ultimately enhance growth and nutrition of important agricultural crops and development of plant-derived products and biofuels

Electroanalysis may be used in the Vanillin Biotechnological Production

This study shows that electroanalysis may be used in vanillin biotechnological production. As a matter of fact, vanillin and some molecules implicated in the process like eugenol, ferulic acid, and vanillic acid may be oxidized on electrodes made of different materials (gold, platinum, glassy carbon). By a judicious choice of the electrochemical method and the experimental conditions the current intensity is directly proportional to the molecule concentrations in a range suitable for the biotechnological process. So, it is possible to imagine some analytical strategies to control some steps in the vanillin biotechnological production: by sampling in the batch reactor during the process, it is possible to determine out of line the concentration of vanillin, eugenol, ferulic acid, and vanillic acid with a gold rotating disk electrode, and low concentration of vanillin with addition of hydrazine at an amalgamated electrode. Two other possibilities consist in the introduction of electrodes directly in the batch during the process; the first one with a gold rotating disk electrode using linear sweep voltammetry and the second one requires three gold rotating disk electrodes held at different potentials for chronoamperometry. The last proposal is the use of ultramicroelectrodes in the case when stirring is not possible

Generation of flavors and fragrances through biotransformation and de novo synthesis

Flavors and fragrances are the result of the presence of volatile and non-volatile compounds, appreciated mostly by the sense of smell once they usually have pleasant odors. They are used in perfumes and perfumed products, as well as for the flavoring of foods and beverages. In fact the ability of the microorganisms to produce flavors and fragrances has been described for a long time, but the relationship between the flavor formation and the microbial growth was only recently established. After that, efforts have been put in the analysis and optimization of food fermentations that led to the investigation of microorganisms and their capacity to produce flavors and fragrances, either by de novo synthesis or biotransformation. In this review, we aim to resume the recent achievements in the production of the most relevant flavors by bioconversion/biotransformation or de novo synthesis, its market value, prominent strains used, and their production rates/maximum concentrations.We would like to thank the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469 unit, COMPETE 2020 (POCI-01-0145FEDER-006684), and BiotecNorte operation (NORTE-01-0145FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020—Programa Operacional Regional do Norte.info:eu-repo/semantics/publishedVersio

receptor analysis and urodynamic outcome

Das von einem anaeroben Bakterium gebildete Botulinumneurotoxin wurde Ende des 18. Jahrhunderts entdeckt und 1990 erstmalig in der Urologie zur Therapie der Detrusor – Sphinkter – Dyssynergie eingesetzt. Hierbei wird der Serotyp Botulinumneurotoxin A (BoNT/A), in den M.detrusor der Harnblase injiziert. Es bewirkt nachweislich die Verbesserung der Kontinenz, eine Abnahme des maximalen Detrusordrucks und die gleichzeitige Zunahme der maximalen Blasenkapazität als auch eine verbesserte Blasencompliance und die Zunahme des Blasenvolumens bis zur ersten Detrusorwelle. Obwohl BoNT/A bereits seit über 20 Jahren in der Urologie genutzt wird, wurde es erst im September 2011 bei Erwachsenen zur Therapie der neurogenen Detrusorhyperaktivität infolge einer stabilen subzervikalen Rückenmarksverletzung oder Multipler Sklerose zugelassen. Neben der Therapie von Erwachsenen, wird BoNT/A auch bei speziellen Erkrankungen im Kindes – und Jugendalter angewendet. Schulte- Baukloh et al. waren 2002 die ersten, die ihre Ergebnisse zur Behandlung von neurogenen Blasenfunktionsstörungen bei Kindern (die Ätiologie war meist eine Meningomyelozele) veröffentlichten. Doch gibt es hierfür noch keine Zulassung. Obwohl eine Wirkung von BoNT/A in vielen Studien bereits bewiesen wurde, ist der Wirkmechanismus von BoNT/A an der glatten Muskulatur noch nicht vollständig geklärt. Bekannt ist, dass nach der Bindung und Internalisierung in das präsynaptische Axon die leichte Kette des BoNT, eine Endoprotease, den Fusionskomplex spaltet und damit die Freisetzung von ACh in den synaptischen Spalt unterbleibt. Es folgt eine schlaffe Lähmung der Muskulatur. Dies scheint aber längst nicht die einzige Wirkungsweise zu sein, denn neuere Studien zeigen auch eine Hemmung der Freisetzung von ATP und Substanz P. Auch ist die Expression von Capsaicin sensitiven TRPV– und purinergen P2X– Rezeptoren nach BoNT/A – Gabe gesenkt. Eindrucksvoll untersuchten Apostolidis et al. den Effekt, dass durch die Injektion von BoNT/A die suburotheliale Expression von TRPV und P2X2 – Rezeptoren gesenkt werden konnte und dies mit einer Reduktion der Drangsymptomatik korrelierte. Nach diesen fortschrittlichen Ergebnissen zu der Wirkungsweise von BoNT/A auf der neurogenen Seite der neuromuskulären Endplatte, interessierten uns jetzt die Veränderungen auf der muskulären Seite der neuromuskulären Endplatte. Das Probandenkollektiv in der von uns durchgeführten Studie bestand aus insgesamt 10 Kindern und Jugendlichen (7 Jungen und 3 Mädchen; Mittelwert 14 Jahre, 3 Monate), die an einer neurogenen Blasenfunktionsstörung erkrankt sind. Trotz maximaler oder supramaximaler Anticholinergikabehandlung wurde bei allen Probanden aufgrund von Detrusordrücken > 40cm – H20 eine Harnblasenaugmentation notwendig. Zuvor wurden bei 7 von 10 Kindern zusätzlich zur anticholinergen Therapie durchschnittlich 3,86 BoNT/ A - Detrusorinjektionen durchgeführt, mit einer Gesamtdosis von durchschnittlich 1143 Units. Sie entsprachen unserer Studiengruppe, die 3 Kinder die nie eine solche Therapie erhalten haben, entsprachen unserer Kontrollgruppe. Das in der Harnblasenaugmentation resezierte Gewebe wurde von uns auf das Rezeptorprofil hin untersucht. Hierfür wurde das Resektat fixiert und geschnitten und die mittels primärem und sekundärem Antikörper angefärbten Schnitte mit konfokalem Lasermikroskop (LSM 5Pascal) analysiert. Es wurden sowohl für die muskarinergen Rezeptoren M2 und M3 als auch für die purinergen Rezeptoren P2X1, P2X2 und P2X3 die durchschnittliche mittlere Pixel – Fluoreszenz – Intensität ermittelt und die Ergebnisse von der Studiengruppe mit denen der Kontrollgruppe verglichen. Neben der Rezeptoranalyse interessierte uns auch das urodynamische Outcome der Studienprobanden verglichen mit der Kontrollgruppe. Hierfür führten wir bei allen Patienten aus der Versuchsgruppe vor und nach der letztmaligen BoNT/ A – Detrusorinjektion, bei der Kontrollgruppe nur einmalig eine standardisierte urodynamische Messung durch. Die Rezeptoranalyse zeigte in der Studiengruppe eine geringere Expression aller untersuchten Rezeptoren nach BoNT/A – Detrusorinjektion gegenüber der Kontrollgruppe. Außer für P2X1 erreichten alle Rezeptoranalysen ein Signifikanzniveau von p < 0,05. Weiterhin konnten wir in unserer Studie zeigen, dass diese Rezeptveränderungen in der Tat mit einer Verbesserung der urodynamischen Befunde korrelierten. Mit dieser Studie wurden erstmalig die Rezeptoren auf der muskulären Seite der neuromuskulären Endplatte untersucht und es konnte erfreulicherweise gezeigt werden, dass BoNT/A signifikant die Expression der Rezeptoren, die für die efferente und die afferente Signalweiterleitung zuständig sind, verändert und einen (indirekten) Einfluss auf die Blasenaktivität hat.Identified 1895, Clostridium botulinum is a gram positive anaerobic bacterium that produces botulinum toxin. The first urological use was in 1990 in patients with detrusor – sphincter – dyssynergia and the first description of detrusor injections was in 1999. Therefore botulinum toxin type A was injected. Since then, several studies have shown an improve of continence, the reduction of maximum detrusor pressure, an increase in compliance, an increase of maximum bladder capacity as well as an increase in reflex volume. Although BoNT/A has been used in Urology for more than 20 years it has not been approved since September 2011 and only for neurogenic detrusor overactivity in patients with multiple sclerosis and in patients with subcervical spinal cord injury. In addition to its use in adults Schulte-Baukloh et al. published 2002 first results of children with neurogenic bladder. Other studies followed, though BoNT/A is not approved for any indications in children yet. Although its significant therapeutic benefits have been widely described, the mechanism of action of BoNT/A in smooth muscle is not yet completely clarified. Research has focused almost exclusive on the presynaptic, neurogenic side of the neuromuscular junction. It is known, that the main effect of BoNT in the striated muscle is that it blocks neuromuscular transmission after docking to its specific receptor (SV2) and being internalized. It then prevents the fusion of the vesicles containing acetylcholine with the presynaptic nerve cell membrane by splitting the membrane – bound fusion protein, the synaptosomal associated protein with a molecular weight of25kD (SNAP-25). Acetylcholine cannot be released as a transmitter and hence there is a flaccid paralysis. We know today that the effects of BoNT/A on the urinary bladder are much more far-reaching and are not fully explained by efferent signal prevention due to blocking of acetylcholine release. At best it partially explains the spasmolysis. However, BoNT/A treatment also significantly reduces the bladder’s sensory functions, and hence the imperative urgency symptoms in patients with OAB. Therefore recent studies show a reduced release of ATP and substance P. Furthermore Apostolidis et al. have shown that treatment of detrusor overacticity with BoNT/A is associated with an effect on the bladder afferent pathway. The number of suburothelial afferents expressing P2X3 and/or TRPV1 was significantly reduced after treatment. After these significant results on the presynaptic, neurogenic side of the neuromuscular junction we were encouraged to analyze the effect of BoNT/A detrusor injections on postsynaptic, muscular receptors. 10 children/adolescents (median age 14yrs., 3 months) with neurogenic detrusor overactivity were included in this prospective, single – blind study. BoNT/A detrusor injections (Ø 3,86) had become necessary for 7 children, because the bladder pressure could not be controlled by other means, despite maximum anticholinergic treatment. 3 children had never received BoNT/A therapy. In all patients urodynamic measurements following the ICS standards were regularly conducted. In all 10 patients the last urodynamic examination revealed a high-risk situation (maximum detrusor pressure > 40cmH2O) that failed conservative treatment, thus the indication for augmentation cystoplastic was given. The tissue was placed in a 4% chilled formaldehyde solution and sent to the laboratory for receptor analysis. The sections were incubated with primary antibodies and coupled secondary antibodies were used for visualization. Specimens were examined using a Pascal 5 confocal laser scanning microscope. The receptor analysis showed a tendency towards downregulation of all examined receptors after BoNT/A injections. Except for P2X1, the reduction in density reached a significant level of p<0.05. The reduction of these receptors probably has an impact on the generated force in the urine bladder und could contribute to the improved urodynamic outcome. This study for the first time provides detailed evidence for post-synaptic muscular receptor changes after BoNT/A injections into the detrusor