August Rauber (1841-1917): from the primitive streak to Cellularmechanik

In the early 19th century Karl Ernst von Baer initiated a new research program searching for the mechanisms by which an egg transforms itself into an embryo. August Rauber (1841-1917) took up this challenge. He considered the phylogenetic principle as the right tool to explain the similitude of embryogenetic processes. In extending Baer's approach, he combined comparative embryology and histology in his studies of avian and mammalian embryos. His earlier work demonstrated that the two-layered chick embryo is a modified gastrula and not a "disc" as Wilhelm His had claimed. From the 1880s onwards, he concentrated on the issue of how the development of germ layers is related to tissue differentiation. To address this, he studied the blastopore, epiblast, primitive streak, teratology and the relative importance of nucleus and cytoplasm in heredity. This paper reconstructs some of Rauber's work and concludes that his observations and reflections constituted a new approach combining embryology and histology with "phylogenetic" reasoning.     

Sercarzian immunology--In memoriam. Eli E. Sercarz, 1934-2009

During his long career as a principal investigator and educator, Eli Sercarz trained over 100 scientists. He is best known for developing hen egg white lysozyme (HEL) as a model antigen for immunologic studies. Working in his model system Eli furthered our understanding of antigen processing and immunologic tolerance. His work established important concepts of how the immune system recognizes antigenic determinants processed from whole protein antigens; specifically he developed the concepts of immunodominance and crypticity. Later in his career he focused more on autoimmunity using a variety of established animal models to develop theories on how T cells can circumvent tolerance induction and how an autoreactive immune response can evolve over time. His theory of "determinant spreading" is one of the cornerstones of our modern understanding of autoimmunity. This review covers Eli's entire scientific career outlining his many seminal discoveries.     

Rat-liver lysosomal sialidase. Solubilization, substrate specificity and comparison with the cytosolic sialidase

Purified liver lysosomes, prepared from rats previously injected with Triton WR-1339, exhibited sialidase activity towards sialyllactose, fetuin, submaxillary mucin (bovine) and gangliosides, and could be disrupted hypotonically with little loss in these activities. After centrifugation, the activities with sialyllactose and fetuin were largely recovered in the supernatant, demonstrating that they were originally in the intralysosomal space. The activities towards submaxillary mucin and gangliosides, on the other hand, remained in the pellet. In the supernatant, activity with fetuin or orosomucoid was markedly reduced by protease inhibitors, suggesting that proteolysis of these glycoproteins may be prerequisite to sialidase activity. The intralysosomal sialidase was solubilized from the mitochondrial-lysosomal fraction of rat liver and partially purified by Sephadex G-200, or Sephadex G-200 followed by CM-cellulose. The enzyme was maximally active at pH 4.7 with sialyllactose as substrate and had a minimum relative molecular mass of 60 000 +/- 5000 by gel filtration; it hydrolyzed a variety of sialooligosaccharides , those containing (alpha 2----3)sialyl linkages being better substrates than those with (alpha 2----6)sialyl linkages. The enzyme failed to attack submaxillary mucin and gangliosides. It was also inactive towards fetuin, orosomucoid and transferrin but capable of hydrolyzing glycopeptides from pronase digest of fetuin. In contrast to the intralysosomal sialidase, the sialidase partially purified from rat liver cytosol by (NH4)2SO4 fractionation followed by chromatography on DEAE-cellulose and CM-cellulose hydrolyzed fetuin and orosomucoid to the extent about half that for sialyllactose. The enzyme was maximally active at pH 5.8 and had a relative molecular mass of approximately 60 000. It also hydrolyzed gangliosides but not submaxillary mucin.     

Adventures in oxygen metabolism

Peter W. Hochachka led a grand life of science adventure and left as his legacy a whole new field--biochemical adaptation. Oxygen was at the core of Peter's career and his laboratory made major contributions to our understanding of how animals deal with variation in oxygen availability in many forms. He analyzed the molecular mechanisms that support facultative anaerobiosis, studied muscle exercise metabolism for high speed flight, swimming and running, investigated mammalian diving on many trips to the Antarctic to study Weddell seals, and probed the metabolic and genetic adaptations that provide optimal hypoxia tolerance for humans residing at high altitudes. His work illuminated both biochemical and physiological mechanisms that are used to optimize aerobic metabolism, to compensate for hypoxic insults, and to conserve energy by strong metabolic rate depression under anoxia. His articles, books and lectures galvanized the field with leading-edge insights and theories and he consistently challenged comparative biochemists to use their unique model systems to explore the range and breadth of animal strategies of biochemical adaptation. Lessons drawn from my training in Peter's laboratory have led me on continuing explorations of adaptations in enzyme function, signal transduction, gene expression, and antioxidant defenses ranging over systems of anoxia tolerance, freezing survival, estivation, and mammalian hibernation.     

Linnaean sources and concepts of orchids

Background

Linnaeus developed a robust system for naming plants and a useful, if mechanical, system for classifying them. His binomial nomenclature proved the catalyst for the rapid development of our knowledge of orchids, with his work on the family dating back to 1737 in the first edition of his Genera Plantarum. His first work devoted to orchids, indeed the first monograph of the family, was published in 1740 and formed the basis for his account in Species Plantarum, published in 1753, in which he gave a binomial name to each species. Given the overwhelming number of orchids, he included surprisingly few – only 62 mostly European species – in Species Plantarum, his seminal work on the plants of the world. This reflects the European origin of modern botany and the concentration of extra-European exploration on other matters, such as conquest, gold and useful plants. Nevertheless, the scope of Linnaeus'' work is broad, including plants from as far afield as India, Japan, China and the Philippines to the east, and eastern Canada, the West Indies and northern South America to the west. In his later publications he described and named a further 45 orchids, mostly from Europe, South Africa and the tropical Americas.

Scope

The philosophical basis of Linnaeus'' work on orchids is discussed and his contribution to our knowledge of the family assessed. His generic and species concepts are considered in the light of current systematic ideas, but his adoption of binomial nomenclature for all plants is his lasting legacy.Key words: Classification, Linnaeus, nomenclature, Orchidaceae, orchids     

Properties of sialidase isolated from Actinomyces viscosus DSM 43798

The cell-bound sialidase of Actinomyces viscosus DSM 43798 was solubilized by mechanical cell disruption and lysozyme treatment. The enzyme was enriched 30,000-fold by cation-exchange chromatography, gel-filtration, and FPLC ion-exchange chromatography, thus obtaining 10 micrograms sialidase protein from 26 g wet cells with a specific activity of 680 U/mg protein. Since sialidase activity was also found in the culture medium, this enzyme was isolated as well, requiring the additional application of FPLC gel-filtration. Both sialidase preparations were apparently homogenous on SDS-PAGE and have similar properties. The substrate specificity of the A. viscosus sialidase was tested with 16 sialoglycoconjugates: The enzyme showed a higher activity with serum glycoproteins than with gangliosides, mucins or sialyllactoses. 4-O-Acetylated N-acetylneuraminic acid was not cleaved from equine submandibular gland mucins or serum glycoproteins in contrast to N-acetyl- and N-glycoloylneuraminic acid. 9-O-Acetyl-N-acetylneuraminic acid was released from bovine submandibular gland mucin, as confirmed by TLC. The sialidase hydrolyses alpha(2----6)-linkages more rapidly than alpha(2----8)- and alpha(2----3)-bonds. Cations, except Hg2+, or chelating agents have no influence on enzyme activity. The sialidase has a relatively high molecular mass of 150 kDa, but consists of only one unit. The enzyme is labile towards freezing and thawing, but can be stored at 4 degrees C in 0.1 M acetate buffer, pH 5.     

Recollections of James Neel

In his long and influential career Neel contributed to almost all aspects of genetic epidemiology from mutation to ethical and philosophical issues. His research spanned North America, Japan, Africa, and Latin America in a fascinating equipoise among clinical, biochemical, epidemiological, and molecular studies that have stimulated hundreds of researchers who enjoyed the controversies he generated as much as the insights he provided. Without exception, we treasure recollections of a high-principled and warm-hearted colleague whose field studies were a model for their generation.     

The protein phosphatases of Drosophila melanogaster and their inhibitors

Protein phosphatases-1, 2A and 2B have been identified in membrane and soluble fractions of Drosophila melanogaster heads. Similarities between Drosophila and mammalian protein phosphatase-1 included specificity for the beta subunit of phosphorylase kinase, sensitivity to inhibitor-1 and inhibitor-2, inhibition by protamine, retention by heparin-Sepharose and selective interaction with membranes. In addition, an inactive form of protein phosphatase-1, termed protein phosphatase-1I, was detected in the soluble fraction that could be activated by preincubation with MgATP and mammalian glycogen synthase kinase-3. Inhibitor-2 partially purified from Drosophila had an identical molecular mass to its mammalian counterpart, and recombined with mammalian protein phosphatase-1 to form a hybrid protein phosphatase-1I. Similarities between Drosophila and mammalian protein phosphatase-2A included preferential dephosphorylation of the alpha subunit of phosphorylase kinase, insensitivity to inhibitors-1 and -2, activation by protamine, exclusion from heparin-Sepharose and apparent molecular mass. A Ca2+-dependent calmodulin-stimulated protein phosphatase (protein phosphatase-2B) that was inhibited by trifluoperazine was identified in the soluble fraction. The remarkable similarities between Drosophila protein phosphatases and their mammalian counterparts are indicative of strict phylogenetic conservation and demonstrate that the procedures used to classify mammalian protein phosphatases have a wider application. Characterisation of the Drosophila phosphatases will facilitate genetic analysis of dephosphorylation systems and their possible roles in neuronal and behavioural plasticity in Drosophila.     

Deglycosylation studies on tracheal mucin glycoproteins

Following several model experiments, conditions were developed for optimal deglycosylation of tracheal mucin glycoproteins. Exposure of rigorously dried material to trifluoromethanesulfonic acid at 0 degree C for up to 8 h results in cleavage of essentially all fucose, galactose, and N-acetylglucosamine, about 80% of the N-acetylneuraminic acid (NeuNAc), and a variable amount of N-acetylgalactosamine (GalNAc), the sugar involved in linkage to protein. Residual N-acetylneuraminic acid is sialidase susceptible and apparently in disaccharide units, presumably NeuNAc2----GalNAc. The remaining N-acetylgalactosamine is mostly present as monosaccharides, and a few Gal beta 1----3GalNAc alpha units are also present; both are cleaved by appropriate enzymatic treatment. The saccharide-free proteins obtained from either human or canine mucin glycoproteins have molecular weights of about 100,000 and require chaotropic agents or detergents for effective solubilization.     

Bifunctional properties and characterization of a novel sialidase with esterase activity from Bifidobacterium bifidum

ABSTRACT

Sialidases catalyze the removal of terminal sialic acid from various complex carbohydrates. In the gastrointestinal tract, sialic acid is commonly found in the sugar chain of mucin, and many enteric commensals use mucin as a nutrient source. We previously identified two different sialidase genes in Bifidobacterium bifidum, and one was cloned and expressed as an extracellular protein designated as exo-α-sialidase SiaBb2. The other exo-α-sialidase gene (siabb1) from the same bifidobacterium encodes an extracellular protein (SiaBb1) consisting of 1795 amino acids with a molecular mass of 189 kDa. SiaBb1 possesses a catalytic domain that classifies this enzyme as a glycoside hydrolase family 33 member. SiaBb1 preferentially hydrolyzes α2,3-linked sialic acid over α2,6-linked sialic acid from sialoglycan, which is the same as SiaBb2. However, SiaBb1 has an SGNH hydrolase domain with sialate-O-acetylesterase activity and an N-terminal signal sequence and C-terminal transmembrane region. SiaBb1 is the first bifunctional sialidase identified with esterase activity.

Abbreviations: GalNAc: N-acetyl-D-galactosamine; Fuc: L-fucose; Gal: D-galactose     

Sialidase significance for cancer progression

Aberrant glycosylation is a characteristic feature of cancer cells. In particular, altered sialylation is closely associated with malignant properties, including invasiveness and metastatic potential. To elucidate the molecular mechanisms underlying the aberrancy, our studies have focused on mammalian sialidase, which catalyzes the removal of sialic acid residues from glycoproteins and glycolipids. The four types of mammalian sialidase identified to date show altered expression and behave in different manners during carcinogenesis. The present review briefly summarizes results on altered expression of sialidases and their possible roles in cancer progression. These enzymes are indeed factors defining cancer malignancy and thus potential targets for cancer diagnosis and therapy.     

In defense of Louis Pasteur: Critique of Gerald Geison's deconstruction of Pasteur's discovery of molecular chirality

Louis Pasteur discovered the phenomenon of molecular chirality, based on his studies of tartrate crystals. His finding remains one of the most important discoveries in the history of chemistry and a fundamentally important chemical phenomenon, with essential implications in biology. In his 1995 book The Private Science of Louis Pasteur, the eminent historian of science Gerald L. Geison (1943‐2001) was highly critical of much of Pasteur's work including his discovery of molecular chirality. The in‐depth analysis provided in this article indicates, however, that the negative assessment of Pasteur's chirality work by Geison is entirely without scientific basis. Criticisms of Pasteur in the book for other “transgressions” in his chirality work, such as supposed influences of his personal biases and stubbornly held a priori notions, misrepresentation of his scientific work in his publications and lectures, and unethical and career‐minded conduct, are also not supported by the evidence. Other troubling features of the book include a broad failure to assure accuracy in a variety of fundamental and important information, including errors in names, dates, events, referencing, indexing, and French‐language text.     

Le dossier Vavilov

Vavilov’s dossier. Gould revived the memory of N.I. Vavilov, a victim of the Stalinian system and misjudged among occidental evolutionists. His contribution is impressive in applied research (phytogeography, his list of world-wide plant resources, a unique collection of germplasms intact and always available) as well as in theoretical research work on artificial selection, immunitary relationships between parasite and plant, the bases of his Law of homologous series in hereditary variations, and centers of origin of cultivated plants. Darwinian concept of natural selection were essential for him, but he considered that the evolutionary changes were not only produced by random variations, but by preset channels, recognising the internal constraints of heredity. His heritage has always been maintained in his Institutes. His Evolutionary theories are now confirmed by molecular genetics and systematics. S.J. Gould was the first to revive Vavilov’s memory and scientific importance. During his studies on the gastropod Cerion Gould recognised the balance between external and internal constraints in Evolution. To cite this article: M. Debrenne, F. Debrenne, C. R. Palevol 2 (2003).     

Why are mammalian alkaline phosphatases much more active than bacterial alkaline phosphatases?

Mammalian alkaline phosphatases are 20-30-fold more active than the corresponding bacterial enzymes even though their amino acid sequences are 25–30% absolutely conserved. In the active-site region there are two noticeable differences between the sequences of the bacterial and mammalian enzymes, in the Escherichia coli enzyme positions 153 and 328 are Asp and Lys, respectively, but in the mammalian enzymes His is observed at both of these positions. Site-specific mutagenesis, genetic and X-ray crystallographic data, which will be summarized here, suggest that the His substitutions at positions 153 and 328 are primarily responsible for the differences in properties between the bacterial and mammalian alkaline phosphatases.     

The bacteriophage, its role in immunology: how Macfarlane Burnet's phage research shaped his scientific style

The Australian scientist Frank Macfarlane Burnet-winner of the Nobel Prize in 1960 for his contributions to the understanding of immunological tolerance-is perhaps best recognized as one of the formulators of the clonal selection theory of antibody production, widely regarded as the 'central dogma' of modern immunology. His work in studies in animal virology, particularly the influenza virus, and rickettsial diseases is also well known. Somewhat less known and publicized is Burnet's research on bacteriophages, which he conducted in the first decade of his research career, immediately after completing medical school. For his part, Burnet made valuable contributions to the understanding of the nature of bacteriophages, a matter of considerable debate at the time he began his work. Reciprocally, it was while working on the phages that Burnet developed the scientific styles, the habits of mind and laboratory techniques and practices that characterized him for the rest of his career. Using evidence from Burnet's published work, as well as personal papers from the period he worked on the phages, this paper demonstrates the direct impact that his experiments with phages had on the development of his characteristic scientific style and approaches, which manifested themselves in his later career and theories, and especially in his thinking regarding various immunological problems.     

Q & A. Carl R. Woese. [interview

Carl R. Woese was born and raised in Syracuse, New York. His undergraduate training was at Amherst College (AB 1950) and graduate work at Yale University (PhD 1953). He is currently the Stanley O. Ikenberry University Professor and Center for Advanced Study Professor of Microbiology at the University of Illinois (Champaign-Urbana), where he has been for the past forty years. He was trained as a biophysicist and molecular biologist. He views himself as a molecular biologist in search of Biology. Consequently, his career has been devoted to using molecular methods to approach evolutionary problems. His most notable accomplishments have been determining the universal phylogenetic tree, through molecular sequence analysis, and the discovery of the Archaea, the so-called ‘third form’ of life. For these he has received numerous awards, including a John D. and Catherine T. MacArthur Award, the Leeuwenhoek Medal 1990 (Netherlands Royal Academy), the Waksman Award (National Academy of Science USA), and the Crafoord Prize (Swedish Royal Academy). At present he works on the evolution of cellular organization.     

Paul Ehrlich: Pathfinder in Cell Biology 1. Chronicle of His Life and Accomplishments in Immunology,Cancer Research,and Chemotherapy1: Paul Ehrlich's Recipe for Success: “Patience,Ability, Money and Luck”

The paper reviews the life of Paul Ehrlich and his biomedical accomplishments in immunology, cancer research, and chemotherapy. Ehrlich achieved renown as an organic chemist, histologist, hematologist, immunologist, and pharmacologist. He disliked the formality of school but managed to excel in Latin and mathematics. His role model was an older cousin, Carl Welgert, who became a lifelong friend. Ehrlich studied medicine at Breslau, Strasbourg, Freiburg, and Leipzig, coming under the influence of Wilhelm Waldeyer, Julius Cohnheim, Rudolf Heidenhein, and Ferdinand Cohn. As a medical student, Ehrlich was captivated by structural organic chemistry and dyes. When he was 23, his first paper was published on selective staining. His doctoral thesis, “Contribution to the Theory and Practice of Histological Staining” contained most of the germinal ideas that would guide his future career. Most of his early work was centered in Berlin at Charité Hospital, where he did pioneering studies on blood and intravital staining, and at Robert Koch's Institute for Infectious Diseases, where he undertook important investigations in Immunology. Ehrlich became an authority on antitoxin standardization and developed the “side-chain theory” of antibody formation for which he was later awarded the Nobel Prize. He became director of an Institute for Experimental Therapy in Frankfurt where he continued research in immunology and carried out routine serum testing. He developed new lines of investigation in cancer research and originated the field of chemotherapy. Using principles developed in his early work with dyes, he successfully treated certain experimental trypanosomal infections with azo dyes. His crowning accomplishment was discovering that the compound Salvarsan could control human syphilis. Ehrlich's legacy in immunology and chemotherapy is discussed and an intimate portrait is drawn of Ehrlich the person.     

Sialidase and malignancy: a minireview

Aberrant sialylation in cancer cells is thought to be a characteristic feature associated with malignant properties including invasiveness and metastatic potential. Sialidase which catalyzes the removal of sialic acid residues from glycoproteins and glycolipids, has been suggested to play important roles in many biological processes through regulation of cellular sialic acid contents. The altered expression of sialidase observed in cancer would, therefore, suggest its involvement in the malignant process. In mammalian cells, three types of sialidase cloned and characterized to date were found to behave in different manners during carcinogenesis. Recent progress in molecular cloning of these sialidases has facilitated elucidation of the molecular mechanisms and significance of these alterations. Herein we briefly describe our own studies on sialidase changes associated with malignant transformation and summarize the topic from both a retrospective and a prospective viewpoint. Sialidases are indeed closely related to malignancy and are thus potential targets for cancer diagnosis and therapy.     

The relation between human lysosomal beta-galactosidase and its protective protein

Cultured skin fibroblasts from patients with the lysosomal storage disease galactosialidosis lack a 54-kDa protein which is a precursor of 32-kDa and 20-kDa proteins, which immunoprecipitate with human anti-beta-galactosidase antiserum. The lack of a 32-kDa "protective protein" results in a combined deficiency of beta-galactosidase and sialidase. The mechanism of protection of lysosomal beta-galactosidase against proteolytic degradation is elucidated by sucrose density gradient centrifugation and immunoprecipitation studies. In normal fibroblasts at the low intralysosomal pH, more than 85% of beta-galactosidase exists as a high molecular weight (600-700 kDa) multimer and about 10% as a monomer of 64-kDa. In mutant cells from galactosialidosis patients, the residual enzyme activity, about 10%, is present as a monomer and no multimer exists. After addition of the 54-kDa precursor form of the protective protein, the density pattern of beta-galactosidase in galactosialidosis cells is normalized. Immunoprecipitation studies after sucrose density gradient centrifugation on homogenate and on purified beta-galactosidase from normal fibroblasts show that the protective protein is associated only with the multimeric form of beta-galactosidase. We propose that intralysosomal protection against proteolysis of beta-galactosidase and sialidase is accomplished by aggregation into a high molecular weight complex consisting of multimeric beta-galactosidase, sialidase, and protective protein. The genetic deficiency of the latter, as in galactosialidosis, results in a rapid degradation of monomeric beta-galactosidase and a loss of sialidase activity.