Proceedings of the Twenty-Fifth Annual Biochemical Engineering Symposium

The Annual Biochemical Engineering Symposium Series started in 1970 when Professors Larry E. Erickson (Kansas State University) and Peter J. Reilly (then with University of Nebraska-Lincoln) got together in Manhattan, KS along with their students for a half-day powwow and technical presentation by their students. Ever since then, it has been a forum for Biochemical Engineering students in the heartland of USA to present their research to their colleagues in the form of talks and posters. The institutions actively involved with this annual symposium include Colorado State University, Kansas State University, Iowa State University, University of Colorado, University of Kansas, University of Missouri-Columbia, and University of Oklahoma. The University of lowa and University of Nebraska-Lincoln have also participated in the conference in recent years. The host institutions for the different symposia have been: Kansas State University (1, 3, 5, 9, 12, 16, 20), Iowa State University (6, 7, 10, 13, 17, 22), University of Missouri-Columbia (8, 14, 19, 25), Colorado State University (II, 15, 21), University of Colorado (18, 24), University of Nebraska-Lincoln (2, 4), University of Oklahoma (23). The next symposium will be held at Kansas State University. Proceedings of the Symposium are edited by faculty of the host institution and include manuscripts written and submitted by the presenters (students). These often include works-in-progress and final publication usually takes place in refereed journals. ContentsPatrick C. Gilcrease and Vincent G. Murphy, Colorado State University. Use of 2,4,6-Trinitrotoluene (TNT) As A Nitrogen Source By A Pseudomonas florescens Species Under Aerobic Conditions. Marulidharan Narayanan, Lawrence C. Davis, and Larry E. Erickson, Kansas State University. Biodegradation Studies of Chlorinated Organic Pollutants in a Chamber in the Presence of Alfalfa Plants. S.K. Santharam, L.E. Erickson, and L.T. Fan, Kansas State University.Surfactant-Enhanced Remediation of a Non-Aqueous Phase Contaminant in Soil. Barry Vant-Hull, Larry Gold, and Robert H. Davis, University of Colorado.The Binding of T7 RNA Polymerase to Double-Stranded RNA. Jeffrey A. Kern and Robert H. Davis, University of Colorado.Improvement of RNA Transcription Yield Using a Fed-Batch Enzyme Reactor. G. Szakacs, M. Pecs, J. Sipocz, I. Kaszas, S.R. Deecker, J.C. Linden, R.P. Tengerdy, Colorado State University.Bioprocessing of Sweet Sorghum With In Situ Produced Enzymes. Brad Forlow and Matthias Nollert, University of Oklahoma.The Effect of Shear Stress ad P-selectin Site Density on the Rolling Velocity of White Blood Cells. Martin C. Heller and Theodore W. Randolph, University of Colorado.The Effects of Plyethylene Glycol and Dextran on the Lyophilization of Human Hemoglobin. LaToya S. Jones and Theodore W. Randolph, University of Colorado.Purification of Recombinant Hepatitis B Vaccine: Effect of Virus/Surfactant Interactions. Ching-Yuan Lee, Michael G. Sportiello, Stephen Cape, Sean Ferree, Paul Todd, Craig E. Kundrot, and Cindy Barnes, University of Colorado.Application of Osmotic Dewatering to the Crystallization of Oligonucleotides for Crystallography. Xueou Deng, L.E. Erickson, and D.Y.C. Fung, Kansas State University.Production of Protein-Rich Beverages from Cheese Whey and Soybean by rapid Hydration Hydrothermal Cooking. Pedro M. Coutinho, Michael K. Dowd, and Peter J. Reilly, Iowa State University.Automated Docking of Glucoamylase Substrates and Inhibitors. J. Johansson and R.K. Bajpai, University of Missouri.Adsorption of Albumin on Polymeric Microporous Membranes.https://lib.dr.iastate.edu/bce_proceedings/1024/thumbnail.jp

Dynamics of protein interactions with new biomimetic interfaces: toward blood-compatible biomaterials

2019 Fall.Includes bibliographical references.Nonspecific blood protein adsorption on the surfaces is the first event that occurs within seconds when a biomaterial comes into contact with blood. This phenomenon may ultimately lead to significant adverse biological responses. Therefore, preventing blood protein adsorption on biomaterial surfaces is a prerequisite towards designing blood-compatible artificial surfaces. This project aims to address this problem by engineering surfaces that mimic the inside surface of blood vessels, which is the only known material that is completely blood-compatible. The inside surface of blood vessels presents a carbohydrate-rich, gel-like, dynamic surface layer called the endothelial glycocalyx. The polysaccharides in the glycocalyx include polyanionic glycosaminoglycans (GAGs). This polysaccharide-rich surface has excellent and unique blood compatibility. We developed a technique for preparing and characterizing dense GAG surfaces that can serve as models of the vascular endothelial glycocalyx. The glycocalyx-mimetic surfaces were prepared by adsorbing heparin- or chondroitin sulfate-containing polyelectrolyte complex nanoparticles (PCNs) to chitosan-hyaluronan polyelectrolyte multilayers (PEMs). We then studied in detail the interactions of two important blood proteins (albumin and fibrinogen) with these glycocalyx mimics. Surface plasmon resonance (SPR) is a common ensemble averaging technique for detection of biomolecular interactions. SPR was used to quantify the amount of protein adsorption on these surfaces. Moreover, single-molecule microscopy along with advanced particle tracking were used to directly study the interaction of single-molecule proteins with synthetic surfaces. Finally, we developed a groundwork for a kinetic model of long-term protein adsorption on biomaterial surfaces. In the first chapter, we thoroughly summarize the important blood-material interactions that regulate blood compatibility, organize recent developments in this field from a materials perspective, and recommend areas for future research. In the second chapter, we report the preparation and characterization of dense GAG surfaces that can serve as models of the vascular endothelial glycocalyx. In the third chapter, we investigate how combining surface plasmon resonance, X-ray spectroscopy, atomic force microscopy, and single-molecule total internal reflection fluorescence microscopy provides a more complete picture of protein adsorption on ultralow fouling polyelectrolyte multilayer and polymer brush surfaces, over different regimes of protein concentration. In the fourth chapter, the interactions of two important proteins from the blood (albumin and fibrinogen) with glycocalyx-mimetic surfaces are revealed in detail using surface plasmon resonance and single-molecule microscopy. Finally, in the fifth chapter, the long-term protein interactions with different biomaterial surfaces are studied with single-molecule microscopy an

Mesoporous Metal Oxide Films

This book, entitled “Mesoporous Metal Oxide Films”, contains an editorial and a collection of ten research articles covering fundamental studies and applications of different metal oxide films. Mesoporous materials have been widely investigated and applied in many technological applications owing to their outstanding structural and physical properties. In this book, important developments in this fast-moving field are presented from various research groups around the world. Different preparation methods and applications of these novel and interesting materials have been reported, and it was demonstrated that mesoporosity has a direct impact on the properties and potential applications of such materials. The potential use of mesoporous metal oxide films and coatings with different morphology and structures is demonstrated in many technological applications, particularly chemical and electrochemical sensors, supercapacitors, solar cells, photoelectrodes, bioceramics, photonic switches, and anticorrosion agents

Spectroelectrochemical Study of the Photoinduced Catalytic Formation of 4,4′-Dimercaptoazobenzene from 4-Aminobenzenethiol Adsorbed on Nanostructured Copper

Surface-enhanced raman scattering (SERS) spectra of self-assembled monolayers of 4-aminobenzenethiol (4-ABT) on copper (Cu) and silver (Ag) surfaces decorated with Cu and Ag nanostructures, respectively, have been obtained with lasers at 532, 632.8, 785, and 1064 nm. Density functional theory (DFT) has been used to obtain calculated vibrational frequencies of the 4-ABT and 4,4′-dimercaptoazobenzene (4,4′-DMAB) molecules adsorbed on model Cu surfaces. The features of the SERS spectra depend on the electrode potential and the type and power density of the laser. SERS spectra showed the formation of the 4,4′-DMAB on the nanostructured Cu surface independently of the laser employed. For the sake of comparison SERS spectra of a self-assembled monolayer of the 4-ABT on Ag surfaces decorated with Ag nanostructures have been also obtained with the same four lasers. When using the 532 and 632.8 nm lasers, the 4,4′-DMAB is formed on Cu surface at electrode potentials as low as −1.0 V (AgCl/Ag) showing a different behavior with respect to Ag (and others metals such as Au and Pt). On the other hand, the surface-enhanced infrared reflection absorption (SEIRA) spectra showed that in the absence of the laser excitation the 4,4′-DMAB is not produced from the adsorbed 4-ABT on nanostructured Cu in the whole range of potentials studied. These results point out the prevalence of the role of electron–hole pairs through surface plasmon activity to explain the obtained SERS spectra.Financial support from Ministerio de Economía y Competitividad (Projects CTQ2013-48280-C3-3-R and CTQ2013-44083-P), Fondos Feder, and the University of Alicante are greatly acknowledged

Biomedical applications of Surface Enhanced Raman Spectroscopy - a step forward to clinical practice

Lo scopo di questo progetto di dottorato \ue8 quello di utilizzare delle superfici metalliche nanostrutturate come substrati per la spettroscopia Raman amplificata da superfici (SERS) per l\u2019analisi di biofluidi. Questa tecnica analitica restituisce l\u2019impronta digitale vibrazionale del campione grazie alla presenza della nanostruttura metallica. Queste caratteristiche anticipano le potenzialit\ue0 della spettroscopia SERS in campo bioanalitico che ha visto un aumento esponenziale delle sue applicazioni nell\u2019ultimo decennio. In particolare, la SERS richiede la fabbricazione di substrati metallici nanostrutturati che possano funzionare da sensori. Questo progetto si basa sullo sviluppo di un approccio privo di marcatura (label-free:): nessuna funzionalizzazione \ue8 presente sulla superficie metallica al fine di rilevare in modo aspecifico gli analiti presenti della matrice di interesse biologico. Il risultato del segnale SERS sar\ue0 un\u2019istantanea della soluzione in analisi depositata sulla superficie metallica, cio\ue8 l\u2019impronta specifica del campione. Per esempio, l\u2019analisi label-free dei biofluidi riflette il suo contenuto metabolico. Nell\u2019era \u201comica\u201d, il SERS pu\uf2 essere integrato nella metabolomica non funzionalizzata in quanto fornisce il profilo metabolico del soggetto in esame e di conseguenza distinguere campioni diversi basandosi sulle differenze di ogni profilo analizzato. I colloidi stabilizzati elettrostaticamente sono stati scelti per la loro nota compatibilit\ue0 con i biofluidi. Verranno usati sia in forma colloidale in sospensione acquosa, sia fissati su un supporto di carta, definiti supporti solidi e sviluppati grazie a un protocollo validato nel nostro laboratorio. Il vantaggio portato dai supporti in carta risiede nella stabilit\ue0 della risposta spettroscopica: sono di lunga durata, facili da fabbricare e da maneggiare, economici e veloci, potenzialmente fabbricabili su ampia scala. Queste sono le caratteristiche che nell\u2019ambito delle applicazioni del SERS possono promuovere la costruzione di un dispositivo Point of Care. Basandosi sulle competenze acquisite dal nostro gruppo di ricerca, lo scopo di questa tesi di dottorato \ue8 duplice: aumentare le nostre conoscenze sull\u2019interazione biofluidi-nanostrutture e utilizzare il metodo SERS per lo studio di specifici problemi clinici. Al fine di soddisfare tali richieste questo lavoro \ue8 diviso in tre parti: 1. Sviluppare protocolli per l\u2019analisi label-free delle frazioni di sangue (siero, plasma, eritrociti, cellule mononucleate del sangue periferico, e sangue intero) con il SERS, sfruttando le loro caratteristiche in base alla diversa preparazione dei campioni e ai substrati SERS utilizzati; 2. Caratterizzare il comportamento delle biomolecole sulla superficie di nanoparticelle metalliche su sistemi modello, cio\ue8 capire il ruolo delle corone di proteine e non proteine nell\u2019interazione metabolita-nanoparticelle. Il sistema modello usato si basa su un insieme di albumina di siero umano (la pi\uf9 abbondante proteina del siero) e molecole che sono comunemente osservate nei biofluidi: adenina, ipoxantina e acido urico; 3. Applicare le nozioni di cui sopra per la diagnosi precoce di diverse malattie (tumore al seno, fegato grasso non alcolico, cirrosi e carcinoma epatocellulare) tramite campioni di sangue e plasma e l\u2019uso di analisi dati multivariata per spettri SERS. Lo scopo dell\u2019utilizzo del SERS in ambito medico \ue8 di proporre nuovi approcci diagnostici complementari alle tecniche gi\ue0 in uso in clinica come ad esempio i metodi di immunochimica e istopatologia. Il vantaggio del SERS risiede nella rapida risposta e in un approccio non invasivo tramite l\u2019utilizzo di biopsia liquida. Lo scopo futuro \ue8 lo sviluppo di una piattaforma SERS label-free come dispositivo point of care integrato allo strumento RamanThis PhD project aims to apply nanostructured metal surfaces as substrates for Surface Enhanced Raman Spectroscopy for the study of biofluids. This analytical technique provides the vibrational fingerprint of a sample assisted by nanostructured metal surfaces, which can enhance the scattering signal of analytes adsorbed on them: this allows detection of analytes in very low concentrations. These features tell a lot about the potential of SERS in the bioanalytics, and indeed, in this field, the use of SERS has increased over the past decade taking advantage of both sensitive detection and fingerprinting features. Above all, SERS requires the manufacturing of metal nanostructured substrates as sensors. In particular, this project is based on the development of a label-free approach: no functionalization is present on the nanoparticles surface, and, hence, no preferential affinity for a given analyte in the biological matrix is sought. Briefly, once nanoparticles are in contact with the specimen, the analytes may adsorb on them without any specific interaction other than their affinity for the metal. The outcoming SERS signal will be a snapshot of what actually reached the metal surface, namely a fingerprint of the sample. For instance, the label-free analysis of biofluids reflect the metabolic content of the fluid itself. In the \u201comic\u201d era, SERS can integrate with untargeted metabolomics and provide the metabolic profile of a specimen and distinguish different samples accordingly, based on differences in such profiles. Silver colloids have been chosen, given that their performances with biofluids are known. They have been used both as colloidal suspension in water, and fixed on a paper support, according to an in-house developed protocol for the fabrication of solid substrates. The coupling of metal nanostructures substrates with SERS acts as actual sensors, able to interact with aqueous environment and detect dissolved analytes. The real advantage of the paper supports lay in the stability of the spectroscopic response: they are long lasting, easy to fabricate and to handle, cost and time effective, prone to scale up. These reasons make them potential Point of Care tool in the frame of SERS applications. The aim of this PhD thesis is twofold: to push forward our fundamental knowledge of the nanostructure-biofluid interaction and to test the feasibility of the application of SERS for specific clinical problems. These goals were pursued in three steps: 1. to develop protocols for the label-free analysis of blood fractions (serum, plasma, erythrocytes, periphereal blood mononuclear cells, and whole blood) with SERS, exploiting their features according to several treatments and SERS substrates; 2. to characterize the behaviour of biomolecules at the interface with metal nanoparticles on model systems, namely to understand the role of the protein and non-protein corona in the metabolites-nanoparticle interaction. The model system used is based on mixture of human serum albumin (i.e. the most abundant serum protein) and molecules which are commonly detected in SERS of biofluids: adenine, hypoxanthine and uric acid; 3. to apply the aforementioned knowledge to the early diagnosis of several diseases (breast cancer, non-alcoholic fatty liver diseases, cirrhosis and hepatocellular carcinoma) through serum and plasma samples by means of multivariate data analysis of SERS spectra. Considering the latter application of SERS in the field of disease diagnosis, the aim is to provide new diagnostic methods complementary to the accepted gold standards such as immunochemistry and histopathology methods. The advantages of SERS lay on the rapid response and on the non-invasiveness of the liquid biopsy approach. As a future goal, the development of SERS platforms as label-free point of care tools integrated to portable Raman instruments could bring the diagnosi

Zinc Oxide Nanostructures: Synthesis and Characterization

The summary should be ca. 200 words; this text will present the book in all promotional forms (e.g. flyers). Please describe the book in straightforward and consumer-friendly terms. [Zinc oxide (ZnO) is a wide band gap semiconductor with an energy gap of 3.37 eV at room temperature. It has been used considerably for its catalytic, electrical, optoelectronic, and photochemical properties. ZnO nanomaterials, such as quantum dots, nanorods, and nanowires, have been intensively investigated for their important properties. Many methods have been described in the literature for the production of ZnO nanostructures, such as laser ablation, hydrothermal methods, electrochemical deposition, sol-gel methods, chemical vapour deposition, molecular beam epitaxy, the common thermal evaporation method, and the soft chemical solution method. The present Special Issue is devoted to the synthesis and characterization of ZnO nanostructures with novel technological applications.