Analysis of Amyloid Using Various Methods

This book explores the world of amyloid, consisting of heterogeneous deposits that have captivated scientists for decades. Amyloid, characterized by its highly organized insoluble protein fibrils exhibiting cross-layered quaternary beta-sheet structures, comprises diverse components such as the P-component, sialic acid, O-acyl sialic acid, and sGAG components. This assembly forms an anisotropic entity that induces birefringence in linearly polarized light, creating a distinctive optical signature.

Initially linked to human disease and tissue damage, the predominant idea that amyloid only harms the host has been challenged. Recent pioneering studies have uncovered the existence of functional amyloid fibers in unexpected domains, from bacteria to mammals.

Advances during the 1960s and 1970s, along with seminal immunohistochemical investigations, gave impetus to immunobiological analysis of amyloid in the 1980s. Notable work, such as the exploration of amyloid fiber staining techniques and the comprehensive study of histochemical properties, have enriched the understanding of this enigmatic deposit.

The book presents a current overview of amyloid research in health and disease, with contributions from the prestigious biennial Amyloid Forum. Written by a distinguished researcher with nearly five decades of experience, the book encompasses five meticulously elaborated chapters.

From human and animal deposits to bacterial formations, isolated fibrils, prions, and amyloid-related neurodegenerative disorders, each category of deposit is given a thorough examination. Innovative topo-optical staining techniques - polarization microscopy, fluorescence microscopy and confocal laser scanning fluorescence microscopy - reveal intricate structures in all species. Coherent structural features are authenticated by various staining reactions and different digestions.

A key segment dissects animal prion fibers, revealing their intricate architecture by topo-optical reactions and uncovering selective arrangements of components.

To complete this, the book confirms the postulated helical structure of the amyloid fiber. This innovative idea is validated by laser scanning confocal fluorescence microscopy, which reveals ordered and helically stacked amyloid protein fibrils in ß-sheet conformation. Associations with chondroitin, heparan sulfate, and looser sphingolipid linkages further unravel the structural complexities of amyloid. Also offers an insightful journey into the realm of amyloid, combining historical knowledge with cutting-edge methodologies, redefining the understanding of this intricate phenomenon.

Prof. Dr. med. Josef Makovitzky, born in Hungary in 1942, is a well-known  figure in the field of amyloid and membrane research. His academic career began at the University of Pécs (Hungary), where he studied medicine from 1962 to 1968.

After completing his studies, he became an assistant doctor at the Institute of Pathology at the University of Pécs under Prof Dr George Romhányi, an amyloid researcher.

Later, he assumed the position of Prosector at the I. Institute of Pathology of the Semmelweis University of Budapest from 1973 to 1976 under Prof. Dr. K. Lapis. His dedication to education and medicine led him to become Deputy Head of the Department of Morphology at the ORFI Hospital in Budapest between 1976 and 1983. Prof. Makovitzky's academic journey took him to Jena, Germany, where he completed his habilitation in 1980 under the direction of Prof. Dr. Günther Geyer, which he defended in Budapest. He subsequently moved to Kiel and Erlangen, where he worked as a physician assistant and scientific assistant at the Institute of Pathology, respectively.

In the following years, he assumed duties as deputy chief physician at the AK Hamburg and visiting professor of pathology at the Martin Luther University Halle/Wittenberg. Prof. Makovitzky's dedication to education extended internationally, holding visiting professorships in Budapest, Pécs, Heidelberg, Freiburg i. Br., Ulm, and Szeged, covering diverse fields from neuropathology to special pathology and clinical pathology. Throughout his career, Prof. Makovitzky's academic contributions remained remarkable. He authored 329 publications, 65 book contributions, 1 monograph, and 3 books. His research interests encompass studies related to amyloid, membran and microscopy techniques.

In particular, his paper "The relevance of Aldehyde-bisulfite toluidine blue reaction and its variants in the submicroscopic carbohydrate research" gained prominence, remaining among the top ten histochemistry for three years. His academic activities have been recognized by his membership in prestigious institutions such as the Hungarian Academy of Sciences and the European Academy of Sciences, Arts, and Letters.

Dr. rer. nat. Thomas R. Appel, born in Germany in 1970, teaches chemistry, physics, and scientific project work at the Lower Saxony Boarding School in Bederkesa (near Bremen). His academic career began at the University of Halle/Saale (Germany), where he studied chemistry from 1990 to 1995.

After his studies, he obtained a Ph.D. in Biophysics at the University of Düsseldorf on the analysis of lipids and carbohydrates in prion amyloid. Later, he did postdoctoral studies and led a research group at the Leibniz Institute of Molecular Biotechnology in Jena (Germany) from 2000 to 2003. Since 2004 he has worked as a teacher and science coordinator at the Lower Saxony Boarding School.

Dr. Appel authored 20 original publications, several book contributions, patents, and public science papers. His research interests shifted from amyloid and analytical techniques to chemistry and physics didactics.

His discovery of a lipid and polysaccharide component in prion amyloid opened new insights into the complex composition of amyloid fibrils. He is a member of the German Chemical Society and the German Society for the Promotion of Teaching Mathematics and Natural Sciences.

Prof. Dr. med. Levente Emody, born in Hungary in 1944, is an expert in the field of microbial pathogenesis. His academic career began at the University of Pécs (Hungary) where he studied medicine from 1963 to 1969.

After his studies, he started as a research assistant at the Department of Microbiology of the University of Pécs and progressed in the academic stages to become a professor and head at the same department. At present, he is professor emeritus at the University of Pécs. His research interest has focussed on infections by Listeria monocytogenes and various representatives of the family of Enterobacteriaceae. He has cooperated with the Institute of Experimental Medicine in Saint Petersburg, the University of Würzburg, the University of Lund, the University of Giessen, the University of Heidelberg, and the University of Freiburg / i.Br.

Research accomplishments: a) Characterisation of the biologic functions of Escherichia coli and Proteus morganii haemolysins, b) Description of the matrix protein binding capacities of human pathogenic Yersinia spp, Helicobacter pylori and Aeromonas salmonicida, c) Description of a new fimbria species (GVVPQ fimbriae) on the surface of Salmonella enteritidis and diarrhoic Escherichia coli. d) Demonstration that resuscitated „viable but non-culturable” Legionella pneumophila cells possess virulence capacity. e) Investigation of virulence functions connected with global regulatory gene sequences in Escherichia coli and Salmonella enteritidis, among others.

The results of the above research have been published in journals like Infection and Immunity, Journal of Bacteriology, Molecular Microbiology, and Applied and Environmental Microbiology.

He is a member of the Editorial Board at the Acta Microbiologica et Immunologica Hungarica (1993-), Hungarian Medical Journal (2024-) and International Journal of Medical Microbiology (2005-2022), Editor of a book on urinary tract pathogens published by Kluwer Academic/Plenum Publishers, New York, 2000.

He started the topo-optical investigation of the bacterial surface with Prof. J. Fischer in the 1970s and later extended this research to bacterial amyloids with Prof. J. Makovitzky, resulting in a chapter of this book.

Dr. Beáta Marianna Kovács, MSc, PhD, born in Hungary, in 1977, graduated in 2001 from Eötvös Loránd University, Faculty of Science, Budapest, MSc in Chemistry, Specialization in Pharmaceutical Research. She received her Ph.D. in 2009 from Szent István University, Budapest, Faculty of Veterinary Science. 
Currently, she is working as a Business Development Manager at a global contract research organization providing support in pharmaceutical research and development.