Mutually exclusive expression of fibronectin and glial fibrillary acidic protein in cultured brain cells

The reported expression of the cell surface-associated, mainly mesenchymal glycoprotein fibronectin by cultured glial cells is in discrepancy with recent work on brain tissue failing to demonstrate any glial or neuronal fibronectin. We have investigated the expression of fibronectin in relation to glial fibrillary acidic protein in cultured human glial and glioma cell lines as well as in cultures derived from newborn rat brain. Using double immunofluorescence technique we found that cells containing glial fibrillary acidic protein do not express fibronectin, and vice versa. The only exception to this rule was the occasional finding of fibronectin at points of cell-to-cell adhesion also in relation to cells containing glial fibrillary acidic protein. The results were also tested by polyacrylamide gel electrophoresis of the culture media of the human cell lines, and by subcultures from the brain of newborn rat, cultures stimulated with dibutyryl cyclic AMP (db-cAMP), and by vinblastine treatment of the cells. The lack of expression of fibronectin in cells containing glial fibrillary acidic protein, a gliospecific cytoskeletal protein, is discussed with reference to glio-mesenchymal interactions and glial markers in vitro.     

Ribonucleotide reductase: A structural study of the dimeric iron site

The iron-containing B2 subunit of ribonucleotide reductase from Escherichia coli has been investigated by Raman spectroscopy. Both the tyrosyl radical-containing native protein and the radical-free protein exhibit a resonance-enhanced Raman band at 500 cm^?1. This band is assigned to an Fe-O vibrational mode arising from an oxygen-containing ligand. The failure to observe any tyrosinate ring modes makes it unlikely that ribonucleotide reductase is an iron-tyrosinate protein and rules out tyrosinate oxygen as a ligand. It is proposed that the 500 cm^?1 band in ribonucleotide reductase is analogous to the 510 cm^?1 Fe-O vibrational mode of methemerythrin and arises from an oxo- or carboxylate-bridge between the antiferromagnetically-coupled Fe(III) ions.     

Induced recovery of a suppressed idiotype by immunization with anti-idiotype

Neonatal Balb/c mice were suppressed forthe H8/T15 idiotype by injection of homologous anti-H8 (D. S. Strayer, D. A. Rowley, and H. Köhler, et al J. Immunol.114, 722, 1975. Six to eight weeks later groups of these suppressed mice were immunized up to three times with isologous anti-H8 raised in Balb/c. After a rest of 4 weeks each group was challenged with R36a vaccine and bleedings were obtained before and after this challenge. All sera were assayed for total anti-PC and H8 idiotype amounts by solid-phase radioimmunoassay. After two-preimmunizations with isologous anti-H8 no increase of the H8 levels was observed though these mice responded to R36a immunization with an increase of total anti-PC antibodies. After the third preimmunization, however, the H8 idiotype was increased in sera taken before and after challenge with R36a. These findings demonstrate that the state of neonatal idiotype suppression can be broken by immunization with complementary anti-idiotype.     

Perchlorate binding to cytochrome c: a magnetic and optical study

1. The effects of perchlorate on cytochrome c have been investigated by 1H and 35Cl NMR, electron paramagnetic resonance and optical spectroscopy. 2. The pK values for the formation and disappearance of the major alkaline conformation were found to be displaced from 9.3 to 8.3 and from 10.4 to 10.9, respectively. The displacement was dependent on the ClO4(-) concentration below 0.1 M. 3. Competition experiments between perchlorate and chloride show that ClO4(-) binds both to the neutral and alkaline forms but with a higher affinity for the latter. The appearance of a new binding site in the alkaline form accounts for the markedly enhanced relaxation rate of 35ClO4(-) in this pH range. Complex formation between cyanide and the alkaline species results in the loss of this binding site, which probably is located close to or within the heme crevice. 4. The neutral form of ferricytochrome c also binds perchlorate strongly as evidence by the unique appearance of a high-spin signal dependent on pH and perchlorate concentration. This signal disappears with the same pK value as the neutral form. The effects of perchlorate on cytochrome c are due to specific binding of this ion.     

Identification of two nuclear polyhedrosis viruses from the cabbage moth, Mamestra brassicae (Lepidoptera: Noctuidae)

Two nuclear polyhedrosis viruses from the cabbage moth Mamestra brassicae found in two geographical areas in Europe have been characterized and compared. These two virus isolates have similar biological activities and have the same host range. The two M. brassicae nuclear polyhedrosis viruses can be distinguished by restriction endonuclease analysis of their DNA. They appear to be distinct but related virus strains.     

Studies on the activation of canine trypsinogens in vitro.

The activation of canine anionic and cationic trypsinogen by enterokinase, trypsin, thrombin, plasmin and extracts from canine granulocytes were studied in vitro. Enterokinase activates both trypsinogens about 1000 times faster than trypsin. The enterokinase-catalyzed activation is not inhibited by the main serum protease inhibitors, alpha-macroglobulin and alpha 1-antitrypsin. alpha-Macroglobulin cannot inhibit the activation of the trypsinogens by trypsin but this reaction is completely inhibited by alpha 1-antitrypsin. The results are discussed in relation to the pathogenesis of acute pancreatitis.     

Purification and characterization of human pancreatic colipase.

Two colipases, named colipase I and colipase II, have been isolated from extracts of human pancreatic gland. The two proteins can be separated by ion-exchange chromatography, isoelectric focusing and slab technique gel electrophoresis. The result of this study indicates that the two colipases, both of which are glycoproteins, have identical amino acid compositions. The pI values were found to be 6.1 for colipase I and 5.8 for colipase II. The different colipases have also been found in human pancreatic juice. The N-terminal amino acid was glycine for both colipase I (gland) and colipase II (juice). Only minor differences were found between the colipases isolated from gland and juice, and colipase I from gland alone was examined in detail.     

Structural studies of the capsular antigen of haemophilus infuenzae type c

The structure of the capsular antigen from Haemophilus influenza type c has been investigated, n.m.r. spectroscopy being the principal method used. It is concluded that the antigen is composed of repeating-units having the following structure:

O-Acetyl groups are present in ～90% of the repeating-units.