Tumor-derived angiogenesis factors from rat Walker 256 carcinoma: an experimental investigation and review

Angiogenesis, the process of developing a hemovascular network, is an essential feature of the growth of solid tumors, and is induced by factors secreted by tumor cells. Assay procedures suitable for the investigation of angiogenesis, and for the screening of angiogenesis factors during purification are reviewed; and a number of reports describing the purification of angiogenesis factors, primarily from the rat Walker 256 carcinoma as starting material, are discussed. Work from the authors' laboratory is also presented. Walker 256 cells grown in large-scale culture were the source of a reproducible and homogeneous source of angiogenic material. Factors secreted by these cells were isolated by a series of chromatographic steps. Ion exchange chromatography on carboxymethyl-Sephadex produced two active fractions, one of which was fractionated into several macromolecular species by lectin affinity and hydrophobic adsorption chromatography. The other gave a high mol.wt, active fraction that was resolved into a low mol.wt, active component and a non-angiogenic but possibly carrier molecule with a mol.wt of 140,000. While none of the angiogenic factors were identified chemically, the results demonstrate the existence of both high and low mol.wt tumor-secreted angiogenic substances, confirming the hypothesis for tumor-induced angiogenesis and predicting potential means to interfere with the process of tumor growth.     

Immunochemical characterization of antibody-coated nanoparticles

A new method using surface plasmon resonance (SPR) through the BIAcore was used to demonstrate the specific interaction between an anti-CD4 monoclonal antibody (IOT4a), adsorbed on poly(methylidene malonate 2.1.2) (PMM 2.1.2) nanoparticles, and the CD4 molecule. The results obtained were compared with the interaction of the same immunonanoparticles with rabbit anti-mouse Fc antibodies. The molar ratio (Fc)/(Fab) was 1, suggesting that the same number of epitopes on the Fc and the Fab fragments were accessible after IOT4a adsorption onto nanoparticles. Comparing the observed association rates of free antibody and antibody adsorbed on nanoparticles, the number of molecules of IOT4a antibody on PMM 2.1.2 nanoparticles was estimated as between 2.6 and 3 per nanoparticle. The properties of the antibody-coated nanoparticles are compatible with their use as antibody-targeted pharmacophores.     

A logical approach to the isolation of lactate dehydrogenase isozyme X from human tests: A general rationale for the isolation of homotetrameric LDH isozymes

Summary Immunoadsorbent and oxamate-Sepharose chromatography were used to isolate electrophoretically homogeneous LDH-X from human testes with a final specific activity of 125 IU/mg and good yields: other applications of this approach are discussed.This work was supported by the Rockefeller Foundation.     

Trypsin in human milk

Human milk trypsin was purified by adsorption chromatography on cellulose-bound 4-aminobenzamidine; its molecular weight was about 24,000 daltons. Its concentration determined by a radioimmunoassay varies between 2.9 and 5.6 micrograms/l.     

Mit Sorptionsmitteln gefüllte Papiere als Dünnschicht-Fertigplatten

Summary Paper loses the properties required for use in partition chromatography with increasing degree of grinding of the cellulose from which the paper is made. For example, paper made from a cellulose mixture consisting of 80% (by weight) Aspa-Birkesulfat and 20% Fichtesulfit adsorbs water to a height of 10 cm after 15 min in the case of a degree of grinding of 14.8°SR (Schoppe-Riegler value) and to a height of only 0.9 cm in the case of a value of 73.3°SR. By adding 50–65% (by weight — based on paper) of adsorbents such as Kieselgel G, Kieselgur G, a mixture of both, Aluminiumoxid G and pure silica gel to the finely ground cellulose mixture in the paper-making process, papers are obtained which adsorb water to a height of 7.6 to 9.3 cm. These filled papers are well suited as thin-layer chromatography plates for adsorption chromatography, as was shown by comparative trials with conventional plates using optical brighteners and 2 solvent systems.

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Herrn Dr.E. Becker zum 65. Geburtstag gewidmet.     

Tumor-derived angiogenesis factors from rat Walker 256 carcinoma: an experimental investigation and review

Summary Angiogenesis, the process of developing a hemovascular network, is an essential feature of the growth of solid tumors, and is induced by factors secreted by tumor cells. Assay procedures suitable for the investigation of angiogenesis, and for the screening of angiogenesis factors during purification are reviewed; and a number of reports describing the purification of angiogenesis factors, primarily from the rat Walker 256 carcinoma as starting material, are discussed. Work from the authors' laboratory is also presented. Walker 256 cells grown in large-scale culture were the source of a reproducible and homogeneous source of angiogenic material. Factors secreted by these cells were isolated by a series of chromatographic steps. Ion exchange chromatography on carboxymethyl-Sephadex produced two active fractions, one of which was fractionated into several macromolecular species by lectin affinity and hydrophobic adsorption chromatography. The other gave a high mol.wt, active fraction that was resolved into a low mol.wt, active component and a non-angiogenic but possibly carrier molecule with a mol.wt of 140,000. While none of the angiogenic factors were identified chemically, the results demonstrate the existence of both high and low mol.wt tumor-secreted angiogenic substances, confirming the hypothesis for tumor-induced angiogenesis and predicting potential means to interfere with the process of tumor growth.This work was supported by funds from the Monsanto Company under Agreements with Harward University and from the U.S. Public Health Service, Contract No. N01-CB-43942. We are particularly indebted to Bernard S. Wildi and Monte C. Throdahl for their advice, cooperation, and encouragement. We also thank Dr Judah Folkman for the initial supply of male mouse submaxillary glands and rat Walker carcinoma conditioned medium, for help with the CAM assays, assays in the rabbit cornea, and many helpful discussions; and R. Bretton, A. Ehrlich, B. Evans, F. Fu, N. Hay and B. Tsokanis for excellent technical assistance.Author to whom correspondence should be addressed.     

Application of hydrophobic chromatography to the fractionation of wheat prolamins]

Hydrophobic chromatography is applied to the fractionation of wheat prolamins. Proteins are separated on "Phenyl Sepharose CL 4B" column. They are eluted by variations of pH and polarity of solvent. Components with the same electrophoretic mobility appear in several chromatographic fractions and gliadin groups, as indicated by Woychick classification, are heterogeneous. This method is excepted to give new information about interaction properties of gluten proteins.     

Platelet–bacterial interactions

Many bacteria are capable of interacting with platelets and inducing platelet aggregation. This interaction may be a direct interaction between a bacterial surface protein and a platelet receptor or may be an indirect interaction where plasma proteins bind to the bacterial surface and subsequently bind to a platelet receptor. However, these interactions usually do not trigger platelet activation as a secondary co-signal is also required. This is usually due to specific antibody bound to the bacteria interacting with FcγRIIa on the platelet surface. Secreted bacterial products such as gingipains and lipopolysaccharide may also be capable of triggering platelet activation.     

绿色气相防锈材料的研究与展望

本文对气相缓蚀剂的原理和发展进程作了系统性阐述.概括了气相缓蚀剂缓蚀机理和分子结构方面的研究现状,并对气相防锈材料的加工成型和应用技术进行了归纳总结,展望了该技术领域内气相缓蚀剂技术的研究方向.     

Trypsin in human milk

Summary Human milk trypsin was purified by adsorption chromatography on cellulose-bound 4-aminobenzamidine; its molecular weight was about 24,000 daltons. Its concentration determined by a radioimmunoassay varies between 2.9 and 5.6 g/l.     

Why was M. S. Tswett’s chromatographic adsorption analysis rejected?

The present paper claims that M. S. Tswett’s chromatographic adsorption analysis, which today is a ubiquitous and instrumentally sophisticated chemical technique, was either ignored or outright rejected by chemists and botanists in the first three decades of the twentieth century because it did not make sense in terms of accepted chemical theory or practice. Evidence for this claim is culled from consideration of the botanical and chemical context of Tswett’s technique as well as an analysis of the protracted debate over Tswett’s chromatographic analysis of chlorophyll between him and Leon Marchlewski, a noted chlorophyll chemist of the period. In this way, the paper expands and amends what it calls the ‘textbook story’ of the early history of chromatography, examples of which may be found in historical notes in many textbooks of chemical instrumental analysis and numerous short articles in chemistry journals. The paper also provides an accessible introduction to the early history of chromatography for historians of science likely to know little or nothing about it.     

Interzelluläre Verbindungen beim Plattenepithelkarzinom der menschlichen Haut in der Gewebekultur

Summary A new medium was developed for tissue-culturing of the human squamous cell carcinoma of the skin. The morphology of the outgrowing tumour cells is described, particularly with regard to the projections of cell surface. There are 2 forms to be differentiated: one which is similar to the desmosomes of the normal epithelium of the skin, and one of intercellular connective strands. The different functions of both forms are discussed with regard to the exchange of cell matter.     

Betel nut residues in archaeological samples of human teeth from the Mariana Islands

Two tetrahydropyridine alkaloids, arecaidine and guvacine that are characteristic of betel nut (Areca catechu L.) have been detected in the deliberately stained labial surface of female teeth excavation on Rota, Mariana Islands. This was accomplished using selected ion monitoring techniques in conjunction with gas chromatography/electron impact-mass spectrometry. These alkaloids were not present on the buccal surface of the teeth and indicate the use of betel nut to effect the staining.     

Alkaline phosphatase binds to collagen; a hypothesis on the mechanism of extravesicular mineralization in epiphyseal cartilage

Affinity chromatography on Sepharose 4B-collagen gels was used to test the affinity of alkaline phosphatase for collagen. Results indicate that alkaline phosphatase of preosseous cartilage binds to collagen probably by electrostatic interactions, this interaction is inhibited by proteoglycan subunits. These results suggest that, in vivo, the formation of a collagen-alkaline phosphatase complex may be a step of the process leading to cartilage calcification.     

Separation of specific fractions of synaptosomes by affinity chromatography

Summary The authors describe a new technique for isolation of specific fractions of synaptosomes, on the basis of their surface glycoproteins, by affinity chromatography using lectin-Sepharose columns.     

Large-scale purification of hepatitis B surface antigen using affinity chromatography

Summary Large-scale purification of hepatitis B surface antigen, applicable to the preparation of potential vaccines for prevention of hepatitis B, is described. The method involves the following steps: precipitation of serum with polyethylene glycol 6000, affinity chromatography on concanavalin A-Sepharose and on -aminononyl-Sepharose, and isopycnic centrifugation.This study was supported by grant 09011 from the National Heart, Lung and Blood Institute. We thank Dr. B. Hollinger for testing HB_sAg samples by a complement fixation test.     

A specific GT1 ganglioside-luteinizing hormone interaction induces conductance changes in lipid bilayers.

A specific interaction was demonstrated between GT1 gangliosides incorporated in bilayer membranes and luteinizing hormone. This interaction would allow the penetration of a hormone subunit in the membrane. The results are discussed in terms of adenylate cyclase activation.     

Electrophoretic profiles of proteins and glycoproteins of chick embryo fibroblasts during development]

The variations of proteins and glycoproteins of Chick embryo fibroblasts are studied during development. This investigation is carried out using polyacrylamide disc gel electrophoresis in SDS. Two glycoproteins of high apparent molecular weight (250,000 and 200,000) undergo quantitative modification: they increase from the 8th to 12th day of development and then remain unchanged to the 16th day. They are cell surface components as suggested by fluorescamine labelling and trypsin sensitivity. The results are discussed in terms of relationship between tumor- and embryo cells.     

Isolation and partial characterization of receptor sites of normal human lymphocytes for lectins]

Glycoproteins of the lymphocyte surface are involved in many membrane mediated events. Their specific carbohydrate determinants might interact with lectins. Purification of macromolecules released from normal human lymphocytes by trypsin was performed using gel filtration and affinity properties for Ricinus sanguineus agglutinin. Some structural and biochemical characteristics are given. Relation to HLA determinants and surface immunoglobulins is discussed.