Scleroglucan as an alternative for animal breeding

Scleroglucan ist ein extrazelluläres ß-Glucan, welches vom filamentös wachsenden Pilz Sclerotium rolfsii produziert wird. Scleroglucan ist ein neutrales Homopolyme, das nur aus Glucose besteht. Durch die Gelpermeationschromatographie gekoppelt an einen Viskositäts- und Brechungsindexdetektor ist es gelungen, über die universelle Kalibrierung Molekulargewichte vom Scleroglucan zu bestimmen. Dabei konnte gezeigt werden, dass der Einfluss von Salzen und Stabilisatoren beachtet werden muss. Durch Ultraschallabbau von nativem Scleroglucan konnten eigene Standards produziert werden, die sich in die universelle Kalibrierung mit Pullulan-Standards (DIN-Pullulan) einfügen lassen. Der Abbau von Scleroglucan ist sowohl thermisch als auch enzymatisch möglich ist. Bei 121 °C über 72 h entstehen jedoch keine verschiedenartigen Molekulargewichtsfraktionen. Die Anwendung von Enzymgemischen zeigte, dass kein Enzym eine reine Endoglucanase-Aktivität besitzt. Es ist jedoch gelungen, mit Hilfe des kommerziell erhältlichen und preisgünstigen Enzyms Cellupract® AS 100 (ca. 5 €/25 kg) der Fa. Biopract (Berlin), eine Fraktion mit definiertem Molekulargewicht zu gewinnen. Primär werden oligomere Scleroglucane mit einem Molekulargewicht von ca. 4.000 bzw. 6.000 g/mol gebildet, die das gleiche Verzweigungsmuster (13C-NMR, Dr. Wray, HZI) wie das native Molekül besitzen. Mit Hilfe von fluoreszenzmarkiertem Scleroglucan konnte in Kooperation mit Prof. Breves (TiHo Hannover) nachgewiesen werden, dass natives Scleroglucan nicht in der Lage ist, die Darmwand des Schweins zu passieren. Aus Untersuchungen von gefriergetrocknetem Darminhalt von Schwein, Rind und Pferd (TiHo Hannover) ging hervor, dass eine Verstoffwechslung des Scleroglucans von der Mikroorganismenpopulation der Tiere abhängig ist. Es erfolgte ein Abbau des Scleroglucans beim Rind und Pferd, jedoch war keine Degradation des Scleroglucans beim Schwein festzustellen. Ein Durchbruch in dem Nachweis der immunstimulierenden Wirkung von degradierten Scleroglucanen ergab sich bei porcinen Dendritischen Zellen (MoDC). In Abhängigkeit vom Molekulargewicht zeigten die MoDC eine verstärkte Reifung (erhöhte Expression der Reifungsmarker CD40 und CD80/86), verminderte Endozytoseaktivität und erhöhte stimulatorische Aktivität in Form von verstärkter Zytokinbildung (Kooperation mit Otto-von-Guericke-Universität,Magdeburg).The filamentous growing fungus Sclerotium rolfsii produces an extracellular ß-glucan named scleroglucan. It is a member of neutral homopolymers and thus the determination of the molecular weight is impeded. To determine the molecular weight, a method has been succeesfully developed using a universal calibration for size exclusion chromatography coupled with viscosity allowing to measure precisely the molecular weight of scleroglucan. It was shown that an influence of salts and stabilizers can be observed. Own reference standards were produced by ultrasonic degradation of nativescleroglucan. The combination of pullulan reference standards and scleroglucan references gave a feasible equation. The degradation of Scleroglucan is possible by thermic, ultrasonic and atack. For 72 h at 121 °C formation of different molecular weight fractions failed only glucoase was released. All enzymes mixtures tested showed combined endo- and exoglucanase activity in different extend. Using the commercially available and inexpensive enzyme Cellupract® AS 100 (about 5 € / 25 kg) from Biopract (Berlin), a defined molecular weight was produced. An oligomeric scleroglucan is primarily formed yielding a molecular weight between 4,000 and 6,000 g/mol with the same branching pattern (13C-NMR, Dr. Wray, HZI) as the native molecule. In cooperation with Prof. Breves (TiHo Hannover) we showed with the help of fluorescence labeling that native scleroglcan can´t pass through the intestinal wall of swine. Tests using lyophilized intestinal contents of swines, cattle and horses (TiHo Hannover) offered that the degradation of Scleroglucan is depending on the microbial population of the animal. Total hydrolysation occured in cattle and horses. Hoewer, in pig Scleroglucan was stable against microbial attack. An immune-stimulating effect by degarded scleroglucan was detected in porcine dendritic cells (MoDC) (cooperation with Otto-von-Guericke-University, Magdeburg). Depending on the molecular weight, increased MoDC maturation (increased expression of maturation markers CD40 and CD80/86), decreased endocytosis activity and stimulatory activity in terms of increased cytokine release was detected

ADME Profiling in Drug Discovery and a New Path Paved on Silica

The drug discovery and development pipeline have more and more relied on in vitro testing and in silico predictions to reduce investments and optimize lead compounds. A comprehensive set of in vitro assays is available to determine key parameters of absorption, distribution, metabolism, and excretion, for example, lipophilicity, solubility, and plasma stability. Such test systems aid the evaluation of the pharmacological properties of a compound and serve as surrogates before entering in vivo testing and clinical trials. Nowadays, computer-aided techniques are employed not just in the discovery of new lead compounds but embedded as part of the entire drug development process where the ADME profiling and big data analyses add a new layer of complexity to those systems. Herein, we give a short overview of the history of the drug development pipeline presenting state-of-the-art ADME in vitro assays as established in academia and industry. We will further introduce the underlying good practices and give an example of the compound development pipeline. In the next step, recent advances at in silico techniques will be highlighted with special emphasis on how pharmacogenomics and in silico PK profiling can enhance drug monitoring and individualization of drug therapy

Die subtalare Arthrorise mit dem Kalix®-Implantat in der Therapie des symptomatischen kindlichen Pes planovalgus - Eine Vergleichsstudie mit der extraartikulären Arthrodese nach Grice-Green

Wir präsentieren die Ergebnisse von 42 Operationen bei kindlichem Pes plano-valgus, die als Arthrorise mit dem Kalix-Implantat oder als Arthrodese nach Grice-Green durchgeführt wurden. Bei 70% der Patienten lag eine ICP vor. Die Auswertung erfolgte klinisch, radiologisch, podoskopisch und pedobarografisch. Die Arthrodese zeigte in 75% gute Ergebnisse. Nach 28,6% aller Arthrorisen musste das Implantat wieder entfernt werden. Schmerzen und Luxationen waren die Probleme. Daher erreichte die Arthrorise nur in 36,9% der Fälle ein gutes Er-gebnis. Die ICP-Patienten schnitten schlechter ab als die idiopathischen und nicht paralytischen Patienten. Wir empfehlen den Einsatz der Arthrorise für idiopathische und nicht paralytische Pes planovalgi mit einem Implantat mit ge-ringer Luxationstendenz. Bei durch ICP bedingten Fehlstellungen wird weiterhin der Einsatz der Methode nach Grice-Green empfohlen. We present the results of 42 surgical interventions for the therapy of pes planovalgus of the child, being done by arthroereisis with the Kalix-implant or by arthrodesis according to Grice-Green. In 70% cerebral palsy was the basic diagnosis. The evaluation was done clinically, radiographically, by podoscope and by pedobarography. The arthrodesis showed good results in 75%. 28.6% of the Kalix-implants had to be removed because of pain or luxation. The arthroereisis therefore showed good results in only 36.9%. The cerebral palsy patients had poorer results then the other patients. We recommend the arthroereisis for idiopathic or non paralytic pes planovalgi with a safe implant. Grice-Green would be the better choice for cerebral palsy patients

Razvoj računalnog upravljačkog sustava za višedjelni fluorescentni detektor

A new control system for fluorescence and EXAFS studies has been developed consisting of germanium detectors and digital signal processor electronics. Using a standard PC and a new software package leads to a high flexibility regarding application, user interface and use of different synchrotron facilities. The new system achieves spectra with considerably higher quality when compared to a traditional system for fluorescence and EXAFS studies. This allows the investigation of thin films and diluted systems as shown by experiments on a W/Nb multilayer system and on FeS2 doped with Zn.Razvili smo upravljački sustav za proučavanje fluorescencije i EXAFS koji se sastoji od germanijskih detektora i signalne digitalne procesorske elektronike. Primjenom standardnog osobnog računala i novog programskog paketa, postigli smo veliku prilagodljivost u primjenama, s međujedinicama i za različite sinkrotronske postave. Nov sustav postiže znatno bolje spektre u usporedbi s uobičajenim sustavima za proučavanje fluorescencije i EXAFS. Tako su moguća istraživanja tankih slojeva i razrijeđenih otopina, kako pokazujemo s višeslojnim W/Nb i sa Zn-punjenim FeS2

Razvoj računalnog upravljačkog sustava za višedjelni fluorescentni detektor

A new control system for fluorescence and EXAFS studies has been developed consisting of germanium detectors and digital signal processor electronics. Using a standard PC and a new software package leads to a high flexibility regarding application, user interface and use of different synchrotron facilities. The new system achieves spectra with considerably higher quality when compared to a traditional system for fluorescence and EXAFS studies. This allows the investigation of thin films and diluted systems as shown by experiments on a W/Nb multilayer system and on FeS2 doped with Zn.Razvili smo upravljački sustav za proučavanje fluorescencije i EXAFS koji se sastoji od germanijskih detektora i signalne digitalne procesorske elektronike. Primjenom standardnog osobnog računala i novog programskog paketa, postigli smo veliku prilagodljivost u primjenama, s međujedinicama i za različite sinkrotronske postave. Nov sustav postiže znatno bolje spektre u usporedbi s uobičajenim sustavima za proučavanje fluorescencije i EXAFS. Tako su moguća istraživanja tankih slojeva i razrijeđenih otopina, kako pokazujemo s višeslojnim W/Nb i sa Zn-punjenim FeS2