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J Blok

Publications and source records attributed to J Blok.

At least 37 records · Page 2Linked to original sources
PubMed1987

Nucleotide sequence comparisons of turnip yellow mosaic virus isolates from Australia and Europe.

The genomic sequences of four isolates of turnip yellow mosaic virus (TYMV-Cd) from Australia, and three TYMV-1 (type) and three TYMV-2 (cauliflower) isolates from Europe were compared by cDNA-RNA hybridization tests, by analysis of the fragments produced from cDNA-RNA hybrids by restriction endonuclease treatment, and by determining the 3' terminal nucleotide sequences of their coat protein mRNAs. All three methods showed only slight differences (ca. 1%) between the mRNA sequences of different TYMV-1 and TYMV-Cd isolates, and did not distinguish between those groups of isolates. By contrast, the nucleotide sequences of TYMV-2 isolates differed from those of the other TYMVs by ca. 5% (sequence analysis) to 11% (restriction fragment analysis). Published biogeographic evidence has indicated that the TYMV-Cd and TYMV-1 populations probably separated more than 12,000 years ago. This implies that these TYMV genomes have changed at a rate of, at most, 1% in 10,000 years.

Australia↗
PubMed1987

The classification of tymoviruses by cDNA-RNA hybridization and other measures of relatedness.

The relationships of twelve tymoviruses have been assessed by cDNA-RNA hybridization. In addition, the percentage molar nucleotide composition of the genome of the PD strain of Kennedya yellow mosaic virus and the percentage molar amino acid composition of the coat proteins of cacao yellow mosaic, Kennedya yellow mosaic and turnip yellow mosaic (Cardamine strain) viruses were estimated. These as well as published serological comparisons and genome and coat protein composition determinations were used to compute classifications of tymoviruses using various "metrics", and simple numerical methods were used to compare the classifications. Measures of relatedness estimated from cDNA-RNA hybridization and base ratio data correlated significantly with each other, but were less closely correlated with those calculated from amino acid data, and did not correlate with those calculated from serological tests. The serological relationships correlated significantly with estimates of relatedness calculated from amino acid data, but not with those based on hybridization or base ratio data. The differences between these classifications mostly resulted from the anomalous behaviour of eggplant mosaic virus, its particles are serologically close to those of other tymoviruses that naturally infect species of the tobacco family, whereas in cDNA-RNA hybridization tests eggplant mosaic virus is closest to the tymoviruses that infect legumes. Similar but smaller anomalies in the characteristics of other tymoviruses were also found.

Amino Acids↗
PubMed1987

An optical study of the conformation of the aminofluorene-DNA complex.

Calf thymus DNA was modified with 2-aminofluorene (AF) to different extents by treatment with N-hydroxy-2-aminofluorene. The AF-modified DNAs together with free AF, the AF-modified guanine (Gua-C8-AF) and the AF-modified deoxyguanosine (dGuo-C8-AF) were subsequently studied by u.v. absorbance, linear dichroism and fluorescence spectroscopy. The emission and absorption properties of double-stranded DNA-AF and single-stranded DNA-AF closely resemble those of dGuo-C8-AF. The emission spectra of these three compounds show a broad, red-shifted emission, characteristic for exciplex formation. The linear dichroism and circular dichroism spectra of double-stranded DNA-AF show that the AF moiety forms a well-defined, regular structure. The dichroic ratio in the 310-340 nm region is constant, which indicates the presence of only one type of adduct. The long-wavelength transition moment of this adduct makes an angle of 72-74 degrees with the DNA helix axis. The binding of AF to double-stranded is DNA is accompanied by a destabilization of the DNA helix structure, a strong quenching of the AF emission quantum yield, intense AF circular dichroism and an apparent immobilization of the dGuo-C8-AF complex. In single-stranded DNA-AF, the AF conformation appears more random, although the interactions between AF and the surrounding bases persist. The strong interactions between AF and the surrounding bases which dominate the optical properties of the studied complexes, the significant destabilization of the DNA double helix after modification with AF, and the relatively small angle between AF and the base planes support a model in which the adduct is inserted into the DNA helix.

Carcinogens↗
PubMed1986

Light and electron microscopic demonstration of osteocalcin antigenicity in embryonic and adult rat bone.

Of the noncollagenous proteins in bone, about 20% consists of osteocalcin. This vitamin K-dependent protein can be found in adult bone, but its presence in embryonic bone could not be demonstrated unequivocally by biochemical methods. Therefore, we used light and electron microscopic immunohistochemical methods to investigate whether osteocalcin antigenicity could be demonstrated in radii of 20-day-old rat embryos. The results show that osteocalcin antigenicity can be demonstrated in the bone matrix of adult bone and in the shaft and endochondral bone matrix of embryonic bone. It could not be demonstrated in calcified cartilage matrix. In bone the antigenicity was observed in the early foci of calcification, i.e., the mineralization nodules.

Animals↗
PubMed1986

Binding stoichiometry of the gene 32 protein of phage T4 in the complex with single stranded DNA deduced from boundary sedimentation.

Short 145 base DNA fragments in complex with the helix destabilizing protein of bacteriophage T4, GP32, have been studied with boundary sedimentation. The sedimentation coefficient was determined as a function of concentration, protein-nucleic acid ratio, temperature and salt concentration. It can be concluded that the measured values reflect the properties of the saturated DNA-GP32 complex. A combination of the earlier obtained translational diffusion coefficient of the complex with the sedimentation coefficient yields its anhydrous molecular weight (Mw = 5.4.10(5) D), which corresponds to a size of the binding site of 10 nucleotides per protein. This procedure is not sensitive to the presence of non-binding protein molecules and to the assumed protein concentration, and therefore, it seems more reliable than a determination from titration experiments. Similar sedimentation measurements were performed with tRNA-complexes containing 76 nucleotides. The translational diffusion coefficient can be calculated from the measured rotational diffusion coefficient and assuming the same hydrodynamic diameter for this complex as obtained for the 145 b DNA complex. The molecular weight derived from the data then also leads to a binding site size of about 10 nucleotides. This suggests that also the short tRNA-complex forms an open, strongly solvated structure, as was proposed for the 145 b DNA-GP32 complex.

Binding Sites↗
PubMed1985

Hydrodynamic studies of a DNA-protein complex. Dimensions of the complex of single-stranded 145 base DNA with gene 32 protein of phage T4 deduced from quasi-elastic light scattering.

The translational diffusion coefficient of the saturated complex of single-stranded 145 base DNA and the helix-destabilizing protein of phage T4, GP32, can be measured at equilibrium by means of quasi-elastic light scattering. If the complex is considered as a rigid rod one can estimate its dimensions by combining the translational diffusion coefficient with earlier data on rotational diffusion. It was found that the average base-base distance of the 145 base DNA in the complex is between 4.3 and 4.7 A, while the diameter of the complex is between 44 and 68 A. This suggests that the conformation of the complex must be such that a large amount of water is trapped.

DNA, Single-Stranded↗
PubMed1985

Hydrodynamic studies of a DNA-protein complex. Elongation of single stranded nucleic acids upon complexation with the gene 32 protein of phage T4 deduced from electric field-induced birefringence experiments.

Short DNA and RNA fragments complexed with the helix destabilizing protein of bacteriophage T4, GP32, have been studied in solution by electric birefringence and circular dichroism. The birefringence of the complexes is positive and the magnitude indicates that the DNA and RNA fragments become linear and rigid upon protein binding. The field free decay is biphasic. On the basis of a rigid rod approximation the slow relaxation time leads to a base-base distance along the helix axis in the complex from 4.3 to 5.6 A, an elongation of at least 50% compared to single-stranded DNA.

Birefringence↗
PubMed1985

Cell surface antigens on osteoclasts and related cells in the quail studied with monoclonal antibodies.

The properties of five monoclonal antibodies raised against isolated osteoclasts are described. Osteoclasts were isolated from medullary bone of egg-laying female quails. Mice were immunized with cell preparations consisting for about 10% of multinucleated osteoclasts. A large number of monoclonal antibodies against cell surface antigens were obtained, five of which were extensively characterized by their interactions with different tissues of the quail and their cross-reactivity with other species. Two monoclonals (OC 5.3 and OC 6.8), recognize surface antigens present on osteoclasts, monocytes, granulocytes and endothelial cells, but not on osteoblasts, osteocytes, fibroblasts, lymphocytes, erythrocytes and others. The three other monoclonal antibodies are specific for multinucleated osteoclasts in bone tissue but recognize some cell surface structures in other tissues. Antibody OC 6.9, which in bone tissue stains primarily the surface area of the osteoclast that is adjacent to the resorbing bone surface, also interacts with bile capillaries in the liver and with specific, but not yet identified parts of the nephron. The antibodies OC 6.1 and OC 6.3 interact with Kupffer cells in the liver and tissue macrophages of small intestine. In view of the possible fallacies inherent to the use of cell surface markers for the demonstration of cell relationship and origin, definite conclusions can not yet be made. The fact that the osteoclast, the Kupffer cell and the intestine macrophage are the only cells in bone, bone marrow, liver, kidney and intestine, that share the same surface antigen recognized by monoclonals OC 6.1 and OC 6.3, suggests, however, a common origin for osteoclasts and a number of well described tissue macrophages.

Animals↗
PubMed1985

The conformation of the complex of the helix destabilizing protein GP32 of bacteriophage T4 and single stranded DNA.

The conformation of single stranded polynucleotides is changed specifically upon binding of the helix destabilizing protein of bacteriophage T4 (GP32). On the basis of circular dichroism (CD) and absorption experiments it is shown that denaturing conditions and the binding of oligopeptides can not induce the altered conformation. On the contrary, according to the current CD and absorption theory, the optical properties of the complex can be explained by a specific, regular conformation, characterized by an appreciable tilt of the bases (less than or equal to -10 degrees) and either a small rotation per base or a small helix diameter. This conformation agrees nicely with the increase of the base-base distance in the complex as determined in solution by electric field induced birefringence measurements. Our calculations show that also the model proposed by Alma (Ph.D. Thesis Catholic University Nijmegen, The Netherlands (1982)) for the complex of the helix destabilizing protein of bacteriophage fd, in which the helix diameter is large and the bases are almost parallel to the helix axis, would agree with the CD- and absorption spectra of the GP32-complex. For the latter protein this model would have to be modified with regard to the axial increment of the bases which is much larger in the GP32-complexes.

Binding Sites↗
PubMed1985

Reaction rate of OH radicals with phi X174 DNA: influence of salt and scavenger.

A derivation is given for the dependence of the rate constant of the reaction of OH radicals with a spherical macromolecule on the rate by which such radicals are scavenged by the medium. Experiments were carried out with oxygenated solutions of dilute single-stranded phi X174 DNA at 10(-4)M NaCl (large reaction radius of DNA) or at 10(-4)M NaCl + MgCl2 (small reaction radius) with t-butanol as a scavenger. The results of these experiments cannot be described by simple second-order competition, but can be explained by the predicted dependence of the rate constant of the reaction OH + DNA on the concentration of t-butanol. Furthermore, the results show that only part of the reactions of OH radicals with phi X174 DNA leads to DNA inactivation, and that even at zero scavenger concentration OH radicals are scavenged by other molecules than DNA, presumably impurities remaining even after careful purification of the DNA.

Bacteriophage phi X 174↗
PubMed1985

Genetic relationships of the dengue virus serotypes.

Previous studies have compared RNA genomes from the different dengue virus serotypes by cDNA-RNA hybridization using dengue-1 virus- and dengue-2 virus-specific cDNA probes. These probes revealed that there is a close genetic relationship between dengue virus serotypes 1 and 4. In this communication, the cDNA-RNA hybridization results using dengue-3- and dengue-4-specific cDNA probes to determine the genetic relatedness of all four dengue virus serotypes are reported. The results indicate that serotypes 1 and 4 are genetically very closely related (sharing about 70% of their genomes as detected by both the dengue-1 and dengue-4 cDNA probes), as are serotypes 3 and 4 (sharing about 50% of their genomes as detected by both the dengue-3 and dengue-4 cDNA probes). Serotype 2 does not seem to be very closely related to the other dengue virus serotypes by cDNA-RNA hybridization analysis.

Base Sequence↗
PubMed1985

Monoclonal antibody PAL-E specific for endothelium.

A monoclonal antibody, PAL-E, is described that is specific for endothelial cells. The monoclonal antibody, an IgG2a, markedly stains endothelium of capillaries, medium-sized and small veins, and venules in frozen sections of human and some animal tissues tested. It reacts not at all or only weakly with endothelium of large, medium-sized, and small arteries, arterioles, and large veins and does not stain the endothelial lining of lymphatic vessels and sinus histiocytes. The cellular staining pattern and tissue staining were different from those obtained with antifactor VIII R:AG antiserum and Ulex europaeus I lectin. Blocking experiments indicated that these three reagents recognize different endothelial binding sites. Therefore, PAL-E is a new staining reagent for endothelium in frozen sections. Based on immunoelectronmicroscopic observations, the antigenic determinant recognized by PAL-E is associated with endothelial vesicles.

Animals↗
PubMed1985

Immuno-electronmicroscopical localization of a microvillus membrane disaccharidase in the human small-intestinal epithelium with monoclonal antibodies.

The cellular localization of the human intestinal disaccharidase, sucrase-isomaltase, was visualized in ultrathin cryosections by the use of specific monoclonal antibodies [25] followed by protein A-gold. The principle site of immunoreaction concerned the microvillus membrane, which supports current concepts of the localization of these hydrolases. One antibody against sucrase-isomaltase also showed labeling of the Golgi apparatus, apical vesicles, and lysosomes, but not of the basolateral membrane. The labeling of the Golgi complex was uniform, suggesting the absence of accumulation of sucrase-isomaltase in cisternae during its passage through this organelle. Absence of labeling of the basolateral membrane appears to support the view that newly synthesized sucrase-isomaltase is transferred directly from the Golgi complex to the microvillus membrane, bypassing the basolateral membrane. However, the results do not exclude the possibility of a very rapid passage through the basolateral membrane. A substantial fraction of the sucrase-isomaltase occurred in lysosomes, which indicates that this organelle plays a major role in the catabolism of microvillar hydrolases. Transport of sucrase-isomaltase to lysosomes might occur by endocytosis or via the crinophagic pathway. The latter was previously postulated to reflect a regulatory mechanism at the post-Golgi level for the surface expression of microvillar membrane proteins.

Antibodies, Monoclonal↗
PubMed1984

Biodegradability test results related to quality and quantity of the inoculum.

For a series of different biodegradation test methods the biodegradation curves are simulated by computer. Simulations are performed on the basis of Monod growth kinetics corrected for cell decay. The possibility of discriminating between growth rates is related to variability of the inoculum quality and quantity. It is concluded that the variability of the inoculum masks all other information. The Repetitive Die Away test, however, offers a good opportunity to obtain information on growth rates that is highly relevant to environmental assessment. The conclusions derived from the computer-simulated curves are confirmed by experimental data.

Bacteriological Techniques↗