Search PubMed⌕ Search

Biomedical subjects

H W Jansen

Publications and source records attributed to H W Jansen.

At least 37 records · Page 2Linked to original sources
PubMed1988

Expression of Met-(-1) angiogenin in Escherichia coli: conversion to the authentic less than Glu-1 protein.

A method for obtaining authentic human angiogenin utilizing an Escherichia coli recombinant expression system is described. A synthetic gene encoding angiogenin was placed into a vector for direct expression under the control of a modified E. coli trp promoter. The protein was produced by the bacteria in an insoluble form and purified to homogeneity by cation-exchange and reversed-phase HPLC following reduction/solubilization and reoxidation. The protein isolated was identified as Met-(-1) angiogenin by amino acid analysis and tryptic peptide mapping; the latter demonstrated that all three disulfide bonds had formed correctly. Both the enzymatic and angiogenic activities of the Met-(-1) protein were equivalent to those of native angiogenin. A Met-(-1) Leu-30 derivative of angiogenin was also isolated and found to be fully active. Conversion of Met-(-1) angiogenin to the authentic less than Glu-1 protein was achieved by treatment with Aeromonas aminopeptidase under conditions in which the new N-terminal glutamine readily cyclizes nonenzymatically. This aminopeptidase treatment may have more general applicability for removal of undesirable N-terminal methionine residues from foreign proteins expressed in bacteria.

Amino Acid Sequence↗
PubMed1988

Morphological aspects of the gingiva in children with Down's syndrome during experimental gingivitis.

In a previous investigation, children with Down's syndrome (DS) showed an earlier, more rapid and more extensive gingival inflammation than normal healthy control children. These differences in gingival inflammation may be the result of aberrant morphology of the gingiva related to the genetic disorder in DS children. The aims of the present study were (i) to describe the structural composition of "normal" gingiva in DS compared to control children, (ii) to analyse the histological changes in the gingiva during plaque development and (iii) to investigate whether the clinical findings could be supported by morphological observations. The study was carried out in 8 DS and 8 matched control children. Their ages ranged from 5-10 years. Gingival normality was guaranteed by strict oral hygiene procedures. During a period of 21 days in which oral hygiene was abolished, gingival biopsies were taken from buccal sites of deciduous teeth following a predetermined schedule on days 0, 7, 14 and 21. Results on day 0 showed no morphological differences between the DS and control children regarding oral epithelium, junctional epithelium or connective tissue. During the experimental phase of the study, the amount of plaque accumulation in the DS children gave rise to a more extensive gingival inflammation than in the control children. The gingival inflammation in the DS group started earlier and included: (1) an acute inflammatory response, (2) an increase of the junctional epithelium area, (3) an increase of the infiltrated connective tissue area (ICT) and (4) a decrease in collagen fibre density of about 35-40% compared to day 0. The same phenomena were not seen until 7 days later in the control group. Conversely, the development of a perivascular lymphocyte infiltrate (LI) in the DS children was delayed compared to the control group. This may be caused by the impaired delayed-type hypersensitivity response in DS children. The development of 2 separate infiltrates (ICT and LI) in this age group and the different temporal development of ICT (day 7 for the DS and day 14 for the control group) and LI (day 14 for the DS and day 7 for the control group) does suggest different immunological mechanisms for both areas and both groups.

Basement Membrane↗
PubMed1988

Cellular aspects of and effects on the gingiva in children with Down's syndrome during experimental gingivitis.

In a previous experimental gingivitis study, it was shown that in children with Down's syndrome (DS), gingival inflammation started earlier, was more extensive and developed faster, than in normal healthy control children. In both groups, the start of the process was accompanied by an acute inflammatory response and an increase of the infiltrated connective tissue area (ICT). The purpose of the present study was to investigate how these facts were reflected at a cytological level. The study was carried out in 8 DS and 8 matched control children. Their ages ranged from 5-10 years. A "normal" healthy gingiva was attained after strict oral hygiene procedures. During a period of 21 days in which oral hygiene was abolished, gingival biopsies were taken on days 0, 7, 14, and 21. In both groups, junctional epithelium (JE) and ICT contained low numbers of polymorphonuclear leucocytes (PMNs). The start of the inflammation (day 7 for the DS and day 14 for the control children) was marked by a significant positive correlation between the numbers of PMNs in the JE and the ICT, and a significant increase of the numbers of PMNs in ICT. In ICT, a concomitant decrease in collagen fibre density was observed. In the control group, the decrease correlated with the numbers of PMNs in ICT, which suggests that this collagen breakdown is caused by PMN products. After the initial decrease, the collagen fibre density remained fairly constant in this group throughout the study. In the DS group, there was a tendency to a further decrease in the ICT3 area, correlated with the numbers of PMNs in ICT.(ABSTRACT TRUNCATED AT 250 WORDS)

Child↗
PubMed1988

Effects of polypeptide growth factors on mandibular condylar cartilage of the rat in vitro.

In a comparative study, the influence of several polypeptide growth factors on proliferation and matrix synthesis in secondary mandibular condylar and primary costal cartilage of the rat were determined using a serum-free culture system. Somatomedin-C, multiplication-stimulating activity (MSA), epidermal growth factor (EGF) and insulin had a significant dose-dependent stimulating effect on proliferation in mandibular condylar cartilage. In costal cartilage, the same factors as well as parathyroid hormone (fragment 1-34), platelet-derived growth factor (PDGF) and high doses of human growth hormone (1 microgram/ml) significantly stimulated proliferation. Matrix synthesis in both cartilages could only be stimulated by high doses of insulin (100 micrograms/ml) and in costal cartilage also by parathyroid hormone. In this culture system fibroblast growth factor reduced proliferation and matrix production, while cartilage-derived factor had no marked effect on the growth processes in both cartilage types. Prominent differences between condylar and costal cartilage were demonstrated by the effects of parathyroid hormone and fetal calf serum. Although the effects on matrix synthesis were very moderate in the tissue culture system used, this study demonstrates that most factors conducive to growth in primary cartilage also stimulate growth in condylar cartilage only exposure to growth substances that interfere with the differentiation of prechondroblasts into chondroblasts, a process that is specific for appositionally growing secondary cartilage, may result in different responses between primary and secondary cartilage.

Animals↗
PubMed1987

Visualization of vascular channels in the mandibular cortex.

Several methods to visualize the vascular pattern of the mandibular cortex were tested. The most suitable method for young jaws proved to be one in which the organic material was partially removed to increase access to the vascular channels. The vessels were vacuum infiltrated with an India ink solution, and the jaws were then decalcified and cleared in glycerin. This resulted in completely transparent flexible cortical plates showing a sharp outline of the vascular pattern. The flexibility of the plates also improved the morphological study of the emergence of blood capillaries at the cortical surface in relation to bone remodeling.

Capillaries↗
PubMed1987

Structure of mutant and wild-type MC29 v-myc alleles and biochemical properties of their protein products.

Proviral DNAs of three fibroblast-transforming MC29 deletion mutants (MC29-10A, MC29-10C, MC29-10H) with defects in hemopoietic cell transformation and tumor induction were molecularly cloned and their deletions were defined by nucleotide sequence analysis. The MC29-10C and MC29-10H v-myc alleles have identical internal deletions overlapping with a smaller one in the MC29-10A v-myc allele, and MC29-10H has an additional internal deletion in the partial gag complement. All deletions are in frame, and the deduced sequences of the mutant gag-myc hybrid proteins lack 56 (MC29-10A) or 120 (MC29-10C, MC29-10H) myc-specific and 44 gag-specific (MC29-10H) amino acid residues. The deleted v-myc nucleotide sequences correspond to the 3' end of exon 2 and the 5' end of exon 3 of the cellular c-myc gene including a region that encodes a high number of acidic amino acid residues. Based on these structural analyses, biochemical properties of mutant and wild-type gag-myc hybrid proteins were compared. Tryptic digests of all three mutant proteins lack a large myc-specific peptide that is present in digests of the wild-type protein and is extensively phosphorylated at serine and threonine residues. Concordantly, the sequence analyses predict that such a large tryptic peptide with putative phosphorylation sites at serine and threonine residues is present in the wild-type gag-myc protein but absent in all three mutant proteins due to the v-myc deletions. Chromatography of wild-type and mutant gag-myc proteins on DNA-cellulose revealed that their in vitro DNA affinities are indistinguishable from each other. Correspondingly, the sequence analyses predict that the carboxyl-terminal region rich in basic amino acid residues and with putative DNA affinity is conserved in wild-type and mutant gag-myc proteins. We conclude that the internal v-myc protein sequences defined by the deletions are necessary for hemopoietic cell transformation and complete phosphorylation, but dispensable for fibroblast transformation and in vitro DNA binding.

Amino Acid Sequence↗
PubMed1986

Nucleotide sequence of the CMII v-myc allele.

The entire nucleotide sequence of the transduced v-myc allele in the genome of avian oncogenic retrovirus CMII was determined. The CMII v-myc and the chicken c-myc alleles differ in their shared coding sequences by a single nucleotide substitution causing a glutamic acid/alanine exchange in the predicted sequences of the corresponding protein products. This mutation has not been found in the transduced v-myc alleles of avian oncogenic retroviruses MC29, MH2, and OK10. We conclude that no specific, if any, missense mutation of the c-myc coding sequence is necessary for oncogenic activation upon transduction of the cellular gene.

Alleles↗
PubMed1986

Interaction between Raf and Myc oncogenes in transformation in vivo and in vitro.

3611 MSV, a raf-oncogene-transducing murine retrovirus, induces fibrosarcomas and erythroid hyperplasia in newborn mice after a latency of 4-8 wk. In contrast, new recombinant murine retroviruses carrying the myc oncogene (J-3, J-5 construct viruses) do not induce tumors before greater than 9 wk. A combination of both oncogenes in an infectious murine retrovirus (J-2) induces hematopoietic neoplasms in addition to less prominent fibrosarcomas and pancreatic adenocarcinoma 1-3 wk after inoculation. The hematologic neoplasms consist of immunoblastic lymphomas of T and B cell lineage and erythroblastosis. If animals were inoculated with a variant of the J-3 virus, which induces altered foci in cultures of NIH 3T3 cells, carcinoma developed in the pancreas with a 2-6 mo latency. In parallel to the synergistic action of both oncogenes on hematopoietic cells in vivo, we find that raf-oncogene-induced transformation of bone marrow cells in culture is enhanced by the addition of myc, which by itself does not transform these cells when grown in standard media. We conclude that concomitant expression of raf and myc oncogenes in hematopoietic and epithelial cells alters their respective transforming activities. The contribution of v-myc in this synergism was examined by use of a series of recombinant murine retroviruses capable of expressing the avian v-myc to study the effect of altered myc expression on hematopoietic/lymphoid cells. With either interleukin 3- or interleukin 2-dependent cell lines, introduction of the recombinant viruses abrogated the requirement for IL 3 or IL 2 for growth, and associated with this was the suppression of c-myc expression. The findings suggest that myc is a component in the signal transduction pathway for IL 3 and IL 2 and support an autoregulatory mechanism of c-myc expression. In contrast to v-myc, expression of v-raf in primary lymphoid/hematopoietic cells has an immortalizing function without abrogating the requirement for IL 3 for growth. This suggests that v-raf and v-myc affect different components of growth regulation, as, for example, commitment (v-myc) and cell cycle progression (v-raf).

Animals↗
PubMed1986

Growth and growth pressure of mandibular condylar and some primary cartilages of the rat in vitro.

To compare the in vitro development of the secondary cartilage of the mandibular condyle with that of primary cartilages, several cartilaginous explants derived from 4-day-old rats were cultured in a serum-free culture system. The following cartilages were used: the mandibular condylar cartilage, the distal epiphyseal cartilage (including the growth plate) of the third metatarsal, a fragment of costal cartilage (including the osteochondral junction) of the fourth rib, the spheno-occipital synchondrosis and the chondroepiphysis of the femoral head. In addition, with a specially designed, in vitro pressure registration system, the maximal growth pressures for each of the explants, except the femoral head, were determined. The results show an independent growth potential for the primary cartilages of the epiphyseal and costal growth plates with a maximal growth pressure of 9.5 and 7.8 g/mm2, respectively. The primary cartilage of the spheno-occipital synchondrosis, on the other hand, although it possesses an independent growth potential, could exert a maximum growth pressure of only 1.5 g/mm2. The secondary cartilage of the mandibular condyle showed a limited intrinsic growth potential, as well as a low maximal growth pressure (2.6 g/mm2). If calculated per dividing and/or matrix synthesizing cell (cells mainly responsible for the cartilage growth), the cells of the condylar cartilage showed the least growth potency (0.08 mg/cell in comparison to 1.9, 1.5 and 0.3 for epiphyseal, costal, and synchondroseal cartilages, respectively.

Animals↗
PubMed1986

Structure and transforming function of transduced mutant alleles of the chicken c-myc gene.

A small retroviral vector carrying an oncogenic myc allele was isolated as a spontaneous variant (MH2E21) of avian oncovirus MH2. The MH2E21 genome, measuring only 2.3 kilobases, can be replicated like larger retroviral genomes and hence contains all cis-acting sequence elements essential for encapsidation and reverse transcription of retroviral RNA or for integration and transcription of proviral DNA. The MH2E21 genome contains 5' and 3' noncoding retroviral vector elements and a coding region comprising the first six codons of the viral gag gene and 417 v-myc codons. The gag-myc junction corresponds precisely to the presumed splice junction on subgenomic MH2 v-myc mRNA, the possible origin of MH2E21. Among the v-myc codons, the first 5 are derived from the noncoding 5' terminus of the second c-myc exon, and 412 codons correspond to the c-myc coding region. The predicted sequence of the MH2E21 protein product differs from that of the chicken c-myc protein by 11 additional amino-terminal residues and by 25 amino acid substitutions and a deletion of 4 residues within the shared domains. To investigate the functional significance of these structural changes, the MH2E21 genome was modified in vitro. The gag translational initiation codon was inactivated by oligonucleotide-directed mutagenesis. Furthermore, all but two of the missense mutations were reverted, and the deleted sequences were restored by replacing most of the MH2E21 v-myc allele by the corresponding segment of the CMII v-myc allele which is isogenic to c-myc in that region. The remaining two mutations have not been found in the v-myc alleles of avian oncoviruses MC29, CMII, and OK10. Like MH2 and MH2E21, modified MH2E21 (MH2E21m1c1) transforms avian embryo cells. Like c-myc, it encodes a 416-amino-acid protein initiated at the myc translational initiation codon. We conclude that neither major structural changes, such as in-frame fusion with virion genes or internal deletions, nor specific, if any, missense mutations of the c-myc coding region are necessary for activation of the basic oncogenic function of transduced myc alleles.

Alleles↗
PubMed1986

Experimental meniscal lesions reconstructed with a carbon fiber-polyurethane-poly(L-lactide) graft.

Reconstructive procedures were investigated in meniscal lesions in 14 dogs. Large wedge-shaped lesions were repaired with a graft containing carbon fibers and an organic polymer, prepared from physical mixtures of poly(L-lactide) and a segmented polyurethane. Four and eight weeks postoperation, arthroscopy was performed and the implants were excised. In ten dogs, the reconstructed area appeared to be invaded almost completely by fibrous tissue and, occasionally, by repair-simulating hyaline cartilage. There were no signs of infection.

Animals↗
PubMed1985

Molecular and biological properties of MH2D12, a spontaneous mil deletion mutant of avian oncovirus MH2.

Avian oncogenic retrovirus MH2 carries two cell-derived oncogenes, v-mil and v-myc. From an infectious stock of MH2 a spontaneous deletion mutant, MH2D12, that has lost most of the v-mil gene but has retained a complete and functional v-myc gene, has been isolated. Nonproducer quail embryo cells transformed by MH2D12 in the absence of helper virus contain two virus-specific proteins: a gag-related protein of 53,000 Da (p53gag), and a v-myc gene product of 59,000/61,000 Da (p59/61v-myc) indistinguishable from the v-myc protein encoded by MH2. MH2D12 viral RNA contains all T1-oligonucleotides specific for the MH2 v-myc gene but none of those characteristic for the v-mil gene. The genetic structure of molecularly cloned proviral DNA of MH2D12 was revealed by restriction mapping, blot hybridization, heteroduplex analysis, and nucleotide sequencing. The MH2D12 provirus is homologous to the MH2 genome but has suffered a deletion of 1271 nucleotides from the central region encompassing the 3' end of delta gag and all of v-mil except the very 3' 31 nucleotides directly adjacent to the v-myc gene. A nine-nucleotide overlap of homology to gag or mil at the delta gag/delta mil junction suggests that recombination between homologous sequence elements of the delta gag and v-mil domains of MH2 was involved in the genesis of MH2D12. The nucleotide sequence analysis predicts that the carboxyterminal 17 amino acids of p53gag are encoded by the residual v-mil sequences and by intron-derived v-myc sequences. Transformation of quail embryo cells by MH2D12 can be assayed by focus and colony formation of transformed cells. This indicates that the v-mil gene is not essential for these activities. However, size and morphology of foci and colonies, and cellular morphology of cultured MH2D12-transformed cell lines can easily be distinguished from those observed in cell transformation by MH2 and resemble more those seen in cell transformation by viruses containing the myc oncogene only.

Amino Acid Sequence↗
PubMed1985

Effects of compressive forces on proliferation and matrix synthesis in mandibular condylar cartilage of the rat in vitro.

The cartilage from 4-day-old rats were exposed to continuous and intermittent compressive forces. Proliferative activity and matrix synthesis in vitro were determined by measuring the incorporation of [3H]-thymidine, [35S]-sulphate and [3H]-proline by autoradiography and liquid-scintillation counting. The findings suggest that a continuously-applied force of approx. 0.5 g stimulated proliferation in the condylar cartilage but reduced the synthesis of the sulphated glycosaminoglycans and collagen. A stimulatory influence on the synthesis of these matrix components, on the other hand, was observed when the condylar cartilage was exposed to an intermittent force (0.7 Hz) of approx. 0.5-1.0 g; this intermittent force, however, reduced the proliferative activity. It appears that, at least in vitro, the basic growth processes in the condylar cartilage can be regulated by compressive forces.

Animals↗
PubMed1985

An in-vitro system for studying the effect of variable compressive forces on the mandibular condylar cartilage of the rat.

An in-vitro system enabled variable range of continuous and intermittent compressive forces to be applied to the cartilage of 4-day-old rats under serum-free culture conditions. A continuous compressive force below 3 g did not affect the growth of the cartilage; when the force exceeded 3 g, growth ceased. Under an intermittent compressive force up to 8 g, growth continued, but at a lower rate; over 8 g, growth ceased. When growth-restricting compression was removed or reduced, growth was reactivated until a new balance was achieved. The normal architecture of the different zones was altered in the non-compressed in-vitro controls, but not in cultured cartilage during the application of these compressive forces.

Animals↗