PubMed Health⌕ Search

Biomedical subjects

C J Jackson

Publications and source records attributed to C J Jackson.

At least 37 records · Page 2Linked to original sources
PubMed1999

Preeclamptic decidual microvascular endothelial cells express lower levels of matrix metalloproteinase-1 than normals.

In preeclampsia, invasion of intrauterine decidual blood vessels by placental cytotrophoblasts is significantly reduced. This study examined the secretion of matrix metalloproteinases (MMP) by cultured human decidual endothelial cells from normal (NDEC) and preeclamptic (PEDEC) pregnancies. MMPs secreted into the culture medium were measured using zymography, Western blotting, and ELISA. Results were confirmed by Northern analysis. Phorbol myristate acetate, known to induce protease activity in other endothelial cell populations, stimulated MMP1, MMP9, and TIMP1 secretion in both NDEC and PEDEC. Neither tumor necrosis factor-alpha nor transforming growth factor-beta, both thought to have significant roles in the control of placentation, affected MMP secretion. MMP9 and TIMP1 levels were similar between the two cell types; however, MMP1 secretion was markedly different between the cell types. NDEC expressed higher levels of MMP1 under both basal (160 +/- 32 ng/10(6) cells) and stimulated (275 +/- 50) conditions compared to PEDEC (32 +/- 24 and 70 +/- 53, respectively). The lower MMP1 expression of decidual endothelial cells from preeclamptic women may inhibit endovascular invasion by cytotrophoblasts. These findings may, at least partly, explain the relative failure of trophoblasts to invade maternal decidual blood vessels in preeclamptic pregnancy.

Capillaries↗
PubMed1999

Species identification and strain differentiation of dermatophyte fungi by analysis of ribosomal-DNA intergenic spacer regions.

Restriction fragment length polymorphisms (RFLPs) identified in the ribosomal-DNA (rDNA) repeat were used for molecular strain differentiation of the dermatophyte fungus Trichophyton rubrum. The polymorphisms were detected by hybridization of EcoRI-digested T. rubrum genomic DNAs with a probe amplified from the small-subunit (18S) rDNA and adjacent internal transcribed spacer (ITS) regions. The rDNA RFLPs mapped to the nontranscribed spacer (NTS) region of the rDNA repeat and appeared similar to those caused by short repetitive sequences in the intergenic spacers of other fungi. Fourteen individual RFLP patterns (DNA types A to N) were recognized among 50 random clinical isolates of T. rubrum. A majority of strains (19 of 50 [38%]) were characterized by one RFLP pattern (DNA type A), and four types (DNA types A to D) accounted for 78% (39 of 50) of all strains. The remaining types (DNA types E to N) were represented by one or two isolates only. A rapid and simple method was also developed for molecular species identification of dermatophyte fungi. The contiguous ITS and 5.8S rDNA regions were amplified from 17 common dermatophyte species by using the universal primers ITS 1 and ITS 4. Digestion of the amplified ITS products with the restriction endonuclease MvaI produced unique and easily identifiable fragment patterns for a majority of species. However, some closely related taxon pairs, such as T. rubrum-T. soudanense and T. quinkeanum-T. schoenlenii could not be distinguished. We conclude that RFLP analysis of the NTS and ITS intergenic regions of the rDNA repeat is a valuable technique both for molecular strain differentiation of T. rubrum and for species identification of common dermatophyte fungi.

Arthrodermataceae↗
PubMed1999

Thrombin rapidly and efficiently activates gelatinase A in human microvascular endothelial cells via a mechanism independent of active MT1 matrix metalloproteinase.

Thrombin has been shown previously to activate gelatinase A in human umbilical vein endothelial cells. The activation is thought to be mediated by membrane-type 1 matrix metalloproteinase (MT1-MMP) on the cell surface, which generates the 62-kd intermediate and the 59-kd fully active forms. We used microvascular endothelial cells derived from human neonatal foreskin to investigate the mechanism of gelatinase A activation by thrombin. Gelatinase A was measured using zymography. Whereas activation by PMA generated both the 62-kd intermediate and the 59-kd fully active forms of gelatinase A after 24 hours, activation by thrombin produced only the 59-kd species rapidly (within 2 hours). Four findings indicate that MT1-MMP was not involved in thrombin-induced activation: (1) there was no up-regulation of MT1-MMP after 2 hours stimulation by thrombin, even though there was activation of gelatinase A; (2) the 62-kd intermediate species was never detected in response to thrombin; (3) tissue inhibitor of matrix metalloproteinase-2 completely prevented gelatinase A activation induced by PMA but not by thrombin; and (4) the metalloproteinase inhibitor 1,10-phenanthroline did not inhibit thrombin-induced activation. Together, these data demonstrate that activation of gelatinase A by thrombin is different from PMA and operating via a pathway independent of MT1-MMP. The ability of thrombin to rapidly and efficiently activate gelatinase A is likely to be a major contributing factor to its potent angiogenic activity.

Cell Membrane↗
PubMed1998

Active and tissue inhibitor of matrix metalloproteinase-free gelatinase B accumulates within human microvascular endothelial vesicles.

Human gelatinase B is involved in tissue remodeling and angiogenesis. It is thought to be synthesized and rapidly secreted as an inactive precursor. In this report, we have shown that human endothelial cells accumulate active forms of gelatinase B in the cytosol. Microvascular but not macrovascular endothelial cells dramatically increased the expression of cytosolic gelatinase B in response to phorbol myristate acetate. Western blotting showed that tissue inhibitor of metalloproteinase-1 (TIMP1) was also present in the cytosol. Whereas gelatinase B was complexed with TIMP1 in the conditioned medium, it existed as a free enzyme in the cytosol, suggesting that the formation of gelatinase B and TIMP1 complex occurs after their secretion. Immunogold electron microscopy revealed that gelatinase B was localized in secretory vesicles which were especially prominent in invading pseudopodia. In contrast, TIMP1 was found throughout the cytoplasm but was not present in the gelatinase vesicles. The accumulation of intracellular activated gelatinase B, ready for rapid release, may facilitate the migration of microvascular endothelial cells during angiogenesis.

Cell Compartmentation↗
PubMed1998

The role of lipogenesis in the development of obesity and diabetes in Israeli sand rats (Psammomys obesus).

Obesity and diabetes in Israeli sand rats, Psammomys obesus, occur with the sequential transition of animals from normal insulin sensitivity to impaired insulin sensitivity, accompanied by increased adiposity, prior to insulin resistance and obesity, in a manner similar to susceptible human populations. The current study was designed to examine the role of de novo lipid synthesis in the development of excessive weight gain in P. obesus. Sand rats were classified at 12 wk of age into three groups: A, normoglycemic normoinsulinemic; B, normoglycemic hyperinsulinemic; C, hyperglycemic hyperinsulinemic, based on glucose and insulin responses in fed sand rats. Body weight, liver weight, white adipose tissue (WAT) mass and food intake were significantly elevated in Group C compared to Group A (P < 0.05). Lipogenic rate was measured by the amount of 3H incorporated into subscapular brown adipose tissue (BAT), epidiymal WAT and liver per hour, from sand rats with and without access to food. No difference in lipogenic rate was found between the groups in BAT, indicating that this tissue is of minor importance in whole body lipogenesis in P. obesus. In the WAT there was a greater lipogenic rate with the development of obesity and hyperinsulinemia (Group B vs. Group A) but no difference in the liver. However, the onset of hyperglycemia (Group C) further stimulated WAT lipogenesis and initiated increased hepatic lipogenesis, both of which contributed to the pre-existing obesity. This study suggests that elevated lipogenesis is not the primary cause of obesity in P. obesus, as lipogenic rate only markedly increases after obesity is already present in hyperglycemic animals.

Adipose Tissue↗
PubMed1998

Associations between heat-stable (O) and heat-labile (HL) serogroup antigens of Campylobacter jejuni: evidence for interstrain relationships within three O/HL serovars.

A comparative examination of the heat-stable (O) and heat-labile (HL) serogrouping results for 9,024 sporadic human isolates of Campylobacter jejuni revealed conserved associations between specific O and HL antigens (O/HL serovars). Forty-nine percent of the isolates which grouped for both O and HL antigens belonged to one of three serovars: O 4 complex/HL 1 (17.9%), O 1/HL 2 (16.8%), or O 50/HL 7 (14.5%). Other common serovars were O 2/HL 4 (8.3%), O 6/HL 6 (8.1%), O 53/HL 11 (4.5%), O 19/HL 17 (3.3%), O 5/HL 9 (3.3%), O 9/HL 9 (3.2%), and O 23/HL 5 (3.1%). These 10 serovars accounted for 83.1% of the serogroupable isolates. A large number of strains (41.3%) could be typed by only one of the two methods or could not be serogrouped (11%). Strains belonging to three serovars, O 2/HL 4, O 50/HL 7, and O 23/HL 5, were further characterized by combining data from expressed features (O/HL serogroups, phage groups, and biotypes) with restriction fragment length polymorphism genotypes. These polyphasic data demonstrated that within each serovar, individual isolates showed substantial conservation of both genomic and phenotypic characteristics. The essentially clonal nature of the three serovars confirmed the potential of combined O and HL serogrouping as a practical and phylogenetically valid method for investigating the epidemiology of sporadic C. jejuni infection.

Animals↗
PubMed1998

Rheumatoid synovial endothelial cells secrete decreased levels of tissue inhibitor of MMP (TIMP1).

OBJECTIVES: Angiogenesis (the formation of new blood vessels) is a major component of the inflammatory pannus in rheumatoid arthritis (RA). Matrix metalloproteinase (MMP) secretion by microvascular endothelial cells is an essential step in angiogenesis. The secretion of MMP1, MMP2, MMP9, and TIMP1 by human microvascular endothelial cells derived from RA synovium (RASE) to normal synovium (NSE) and neonatal foreskin (FSE) was compared. METHODS: Confluent monolayers of endothelial cells in basal medium were pre-incubated for 24 hours in the presence or absence of phorbol myristate acetate (PMA, 100 ng/ml). MMP1 activity was measured using a spectrophotometric assay and western blotting. MMP2 and MMP9 were measured using zymography. TIMP1 was measured by enzyme linked immunosorbent assay and western blotting. RESULTS: There was little difference between the amounts of MMP2 secreted by any of the cell lines. In response to PMA both synovial cell types showed a significantly higher MMP1 and MMP9 activity compared with FSE, although there was no difference between RASE and NSE. Tumour necrosis factor alpha had minimal effect on MMP activity. There was a striking decrease in the amount of TIMP1 secreted by RASE compared with normal synovium. CONCLUSIONS: As overall MMP activity is a balance between the amount of MMP and TIMP1 present, the low levels of TIMP1 produced by RASE would shift the balance in favour of increased MMP activity by these cells. This is likely to contribute to the angiogenic potential of RASE.

Arthritis, Rheumatoid↗
PubMed1997

Human microvascular endothelial cells differ from macrovascular endothelial cells in their expression of matrix metalloproteinases.

Matrix metalloproteinase (MMP) secretion by microvascular endothelial cells is an essential first step in the formation of new blood vessels (angiogenesis). Since angiogenesis does not occur in large blood vessels, we investigated whether the secretion of MMPs and tissue inhibitor of MMP (TIMP1) differs between micro- and macro-vascular endothelial cells. We compared the secretion of MMPs and TIMP1 by human endothelial cells derived from neonatal foreskin (FSE) and umbilical vein (HUVE) sources. The cells were incubated for 24 hr in the presence or absence of the angiogenic agents, phorbol myristate acetate (PMA, 100 ng/ml) or tumour necrosis factor-alpha (TNF, 100 ng/ml). The cell supernatants were removed and assayed for MMPs and TIMP1 using a spectrophotometric assay for MMP1, zymography, Western blotting and Northern analysis. When endothelial cells were incubated in basal medium for 24 hr they secreted MMP1, MMP2 and TIMP1 but not MMP9. HUVE secreted substantially higher levels of these proteins compared to FSE. In addition, HUVE secreted two low molecular mass bands representing activated forms of MMP2. These activated forms were not present in supernatants derived from FSE. In response to PMA, both FSE and HUVE increased secretion of MMP1 and TIMP1. However, there was a dramatic difference in level of response by the two cell types with FSE secreting substantially more TIMP1 and MMP9 compared to HUVE. These data clearly show that cultured endothelial cells derived from microvascular vs macrovascular tissues exhibit different MMP and TIMP secretory profiles.

Cells, Cultured↗
PubMed1997

Genotype analysis of human blood isolates of Campylobacter jejuni in England and Wales.

Genomic profiles were obtained for 76 strains of Campylobacter jejuni isolated from bacteraemic patients in England and Wales over the period 1981-94. Genotyping was performed by restriction fragment length polymorphism (RFLP) analysis using a random cloned DNA probe, and by ribotyping with a PCR-generated C. jejuni 16S ribosomal DNA probe. Phenotypic characterization was achieved by heat-stable (HS) and heat-labile (HL) serogrouping, and Preston phagetyping and biotyping. The blood isolates were genomically heterogenous, with 24 RFLP/16S profiles occurring within the 76 strains. Forty-four percent of isolates belonged to one of three RFLP/16S genotypes, reflecting the patterns seen in faecal isolates, except that genotypes usually associated with the HS 1 antigen were uncommon. The two most prevalent genotypes, characteristic of HS 2 and HS 4 strains, showed similarity by cluster analysis. Further evidence was seen of associations between phenotypic and genotypic characters within some HS serogroups. Chromosomal profiling by RFLP analysis does not indicate that particular genotypes have a predisposition to invade the bloodstream.

Bacteremia↗
PubMed1996

The application of genotyping techniques to the epidemiological analysis of Campylobacter jejuni.

Campylobacter jejuni serogroup reference strains and collections of sporadic and outbreak-associated isolates were examined for restriction fragment length polymorphisms (RFLPs), using C. jejuni random chromosomal and 16S rRNA gene probes. A collection of 48 Penner (HS) and 14 Lior (HL) serogroup reference strains, plus 10 clinical isolates, generated 35 RFLP and 26 ribotype patterns. In combination the two loci generated 48 distinct genotypes. Both probes were able to differentiate between certain random isolates of the same HS/HL serogroups but greater discrimination was obtained with RFLP than with ribotyping. Genotyping distinguished accurately between related and unrelated strains when applied to several outbreaks. Genotypic analysis of C. jejuni by restriction fragment length polymorphisms is a valuable technique for epidemiological typing. Chromosomal variation detected by the two unlinked probe loci provides some information about the genetic relationship between isolates.

Campylobacter Infections↗
PubMed1996

A novel polymerase chain reaction assay for the detection and speciation of thermophilic Campylobacter spp.

A novel PCR amplification method is described which is specific for the thermophilic, enteropathogenic species Campylobacter jejuni, Camp. coli and Camp. upsaliensis. Rapid, accurate speciation of amplified strains is possible on the basis of restriction fragment length polymorphisms of PCR products digested with three restriction enzymes, AluI, DdeI and DraI. The sensitivity of detection is 25 cfu in water, and 2 x 10(3) cfu in full cream milk. An epidemiological application of the assay in detecting non-culturable campylobacters from a contaminated potable water supply is described.

Campylobacter↗
PubMed1995

Direct milk excretion of Campylobacter jejuni in a dairy cow causing cases of human enteritis.

Consumption of milk contaminated with Campylobacter jejuni has been described as a cause of human enteritis. Although faecal contamination of milk with the organism has frequently been described, direct milk excretion of Campylobacter jejuni into milk has rarely been linked with cases of human infection. We describe the investigations undertaken following the isolation of Campylobacter jejuni from samples of unpasteurized milk prior to retail. Results of epidemiological investigations including typing of Campylobacter jejuni isolates using pyrolysis mass spectrometry, Penner and Lior serotyping, biotyping, phage typing and restriction fragment length polymorphism analysis provided convincing evidence implicating direct milk excretion of Campylobacter jejuni by one asymptomatic dairy cow as the source of the milk contamination and the cause of local cases of human enteritis.

Animals↗
PubMed1994

VLA-2 mediates the interaction of collagen with endothelium during in vitro vascular tube formation.

A confluent endothelial monolayer can be induced to form vascular tubes in response to collagen. We investigated possible mechanisms of collagen-induced tube formation by using antibodies to the VLA-2 integrin receptor and protein kinase C inhibitors. Pre-incubation of cells with anti-VLA-2 (which recognises both the alpha 2 and beta 1 chains) and AK7 (which recognises only the alpha 2 chain) showed a dose-dependent inhibition of tube formation. At 50 micrograms/ml, anti-VLA-2 completely inhibited collagen-induced tube formation, whereas AK7 caused only partial inhibition. Both chlorpromazine and trifluoperazine, at concentrations of 10 microM, prevented tube formation (> 40% inhibition). In summary, the VLA-2 integrin receptor plays a role in the induction of tube formation by type I collagen. Protein kinase C may be activated during this process.

Amino Acid Sequence↗
PubMed1994

Sulfated polysaccharides are required for collagen-induced vascular tube formation.

We have previously shown that soluble type I collagen can induce vascular tube formation when it contacts the apical side of a confluent endothelial monolayer. In this study we have examined which soluble agent(s) are required for collagen-induced tube formation. Human neonatal foreskin microvascular endothelial cells, maintained in basal medium, were preincubated with each test agent for 2 h prior to the addition of solubilised type I collagen (100 micrograms/ml). After 6 h, tube formation was quantitated using image analysis and expressed as the mean area of tube formation (mm2) per microscopic field of view. Collagen-induced tube formation did not occur in the presence of endothelial cells growth supplement, basic fibroblast growth factor, or normal pooled human serum. In contrast, the addition of heparin at 5 or 50 micrograms/ml caused extensive tube formation (0.22 +/- 0.07 and 0.30 +/- 0.12 mm2, respectively) whereas at 500 micrograms/ml little tube formation occurred (0.03 +/- 0.02 mm2). Protamine sulfate, an antagonist of heparin, inhibited collagen-induced tube formation in a dose-dependent manner. Pentosan polysulfate, dextran sulfate, heparan sulfate, and chondroitin sulfate mimicked the action of heparin. Partially sulfated heparin (de-N-sulfated heparin) stimulated less tube formation compared to heparin (0.15 +/- 0.06 mm2 at 50 micrograms/ml). The nonsulfated polysaccharides, xylan and dextran, had no effect on tube formation. In summary, sulfated polysaccharides are required for collagen-induced vascular tube formation in vitro. The sulfation of these molecules appears to be vital for collagen-induced tube formation.

Cells, Cultured↗
PubMed1992

Possible mechanisms of type I collagen-induced vascular tube formation.

UNLABELLED: We investigated the effect of type I collagen on endothelial behaviour following its contact with the apical versus basal surface of cultured human endothelial cells. When endothelial cells were plated onto type I collagen they attached via their basal surface and formed a confluent monolayer. However, when type I collagen (100 micrograms/ml) was added directly to the growth medium, so that it made contact with the apical surface of endothelial cells, it induced rapid capillary-like tube formation. Possible mechanisms were assessed using a) polyclonal (anti-VLA-2) and monoclonal (AK7) antibodies to different epitopes on the alpha 2 beta 1 integrin receptor for collagen and b) drugs (chlorpromazine and trifluoperazine) that inhibit protein kinase C activity. Both anti-VLA-2 and AK7 (1-50 micrograms/ml) showed a dose-dependent inhibition of tube formation and cell attachment. At 50 micrograms/ml, anti-VLA-2 completely inhibited tube formation whereas AK7 caused only partial inhibition (less than 50%). By contrast, AK7 was a more potent inhibitor of cell attachment than anti-VLA-2. Both chlorpromazine and trifluoperazine prevented tube formation. CONCLUSIONS: 1) The alpha 2 beta 1 integrin receptor plays a role in both endothelial cell attachment and the induction of tube formation by type I collagen. 2) Protein kinase C may be involved in collagen-induced tube formation.

Cell Adhesion↗