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M Hoyle

Publications and source records attributed to M Hoyle.

At least 19 recordsLinked to original sources
PubMed2004

Cytokine response profiles predict species-specific infection patterns in human GI nematodes.

This study investigated associations between pre-treatment cytokine expression and infection patterns, before and after de-worming, in humans exposed to two gastrointestinal nematode species. Quantitative measures of Ascaris lumbricoides and Trichuris trichiura infection (based on faecal egg counts) were estimated immediately before and 8-9 months after treatment in a Cameroonian population. Whole blood cytokine responses to parasite-derived antigens were assayed immediately pre-treatment. An overall measure of the tendency towards species-specific infection (increasing with A. lumbricoides faecal egg counts and decreasing with T. trichiura faecal egg counts) was significantly positively related to IL-10 levels in older (14-57 year) hosts. There was a significant negative influence of IL-5 on reinfection probability in T. trichiura but not A. lumbricoides. This effect coincided with reduced reinfection success in T. trichiura compared to A. lumbricoides. T(H)2 cytokine expression by younger hosts (4-13 year) was negatively associated with contemporary A. lumbricoides faecal egg counts before treatment. Following treatment, the pre-treatment T(H)2 cytokine expression data for younger hosts (now reflecting responsiveness 8-9 months in the past) were negatively associated with T. trichiura faecal egg counts. Taken together, these observations suggest a successional interaction between T(H)2-driven immune responses and species infection over time. However, any differential effects of the measured immune responses on species-specific recruitment, maturation and mortality were superimposed upon (and outweighed by) the effects of other factors favouring coinfection.

Adolescent↗
PubMed1992

Expression of porcine complement cytolysis inhibitor mRNA in cultured aortic smooth muscle cells. Changes during differentiation in vitro.

Porcine smooth muscle cells (SMC) grown to a high density monolayer culture undergo a morphological transition in which the cells draw away from the substrate and form multicellular nodules. The cells within the nodule resemble SMC in the aortic media and in some atherosclerotic plaques. The process of nodule formation is associated with the enhanced production of a secreted 38-kDa glycoprotein. To characterize the 38-kDa protein and its expression, a cDNA clone (pc38K) was isolated by immunological screening of an expression library. The 1646-base pair cDNA contains a single open reading frame encoding 446 amino acids. This sequence shows 72% homology with the human complement cytolysis inhibitor (CLI), also called serum protein-40,40, and 68% identity with rat sulfated glycoprotein-2. Based on this homology, we refer to the protein encoded by pc38K as CLI. This polypeptide includes a potential signal sequence, seven glycosylation sites and 10 cysteines in two clusters of five each. Southern blot analysis reveals that a single copy gene encoding CLI is present in mammals and chicken. In Northern blot analysis of SMC RNA, pc38K hybridizes to a mRNA of about 1.9 kilobases that is preferentially expressed in nodular SMC. The steady state level of this mRNA increases as the cultures begin to form multilayered regions. High levels of the mRNA persist after the cells are trypsin-dissociated. Culture medium conditioned by nodular SMC also induces an increase of CLI mRNA. Analysis of RNA extracted from porcine tissues show the highest levels of CLI mRNA in brain and liver; lower levels are detected in other tissues, including the aorta. Possible functions for the CLI are discussed.

Amino Acid Sequence↗
PubMed1992

Differential expression of metalloproteinase and tissue inhibitor of metalloproteinase genes in aged human fibroblasts.

The basal levels of mRNAs encoding two metalloproteinases, collagenase and stromelysin, were increased as a function of in vitro serial subcultivation (cellular aging) of human fibroblasts. Procollagenase and prostromelysin synthesis and secretion were also greater in the old cultures (late passage). In contrast, the steady-state expression of mRNA for an inhibitor of metalloproteinases, tissue inhibitor of metalloproteinase-1 (TIMP-1), in late-passage cultures was lower than that in young cell cultures (early passage). Each mRNA was analyzed using total RNA preparations isolated from normal fibroblast cultures at different phases of the in vitro life span and from cultures derived from donors with the premature senescence syndromes characterized as Werner syndrome, progeria (Hutchinson-Gilford) syndrome, or Cockayne syndrome. In normal cell cultures expression of metalloproteinase mRNAs was increased after the culture had completed greater than 90% of the in vitro life span, and the reduction in TIMP-1 mRNA expression occurred after the culture had completed greater than 74% of the in vitro lifespan. In Werner syndrome cultures expression of metalloproteinase and TIMP-1 mRNAs was similar to the level of expression observed in late-passage cell cultures. Levels of metalloproteinase and TIMP-1 mRNA expression in progeria and Cockayne syndromes were similar to those of early-passage cell cultures. To determine if young and old cells were each responsive to mediators of metalloproteinase synthesis, cultures were treated with phorbol ester or cytokines. 12-O-tetradecanoylphorbol-13-acetate treatment increased the steady-state levels of all three mRNAs in young, old, and Werner syndrome cultures and increased procollagenase levels in all cultures. Early- and late-passage cell cultures also responded to cytokines. Interleukin-1 alpha treatment increased collagenase and stromelysin mRNA levels while transforming growth factor-beta reduced the steady-state levels of both transcripts. Neither cytokine affected the steady-state level of TIMP-1 mRNA. The results indicate that in vitro cellular aging is associated with changes in expression of mRNAs encoding proteins that mediate inflammatory responses and connective tissue remodeling.

Aging↗
PubMed1989

Collagenase production by early and late passage cultures of human fibroblasts.

Comparison of the proteins secreted by early and late passage cell cultures of human fibroblasts revealed a high level of immunoreactive collagenase (Mr = 55,000 Da and 58,000 Da) in the late passage cell culture conditioned medium. Both molecular weight species reacted with a monoclonal anticollagenase antibody and were apparently glycosylation varaents of the same protein. The question of whether the apparent age-dependent differences in collagenase synthesis reflected changes in protein synthesis or secretion was addressed by assaying immunoreactive collagenase and collagenase mRNA. Immunofluorescence microscopy of cellular collagenase revealed that the percentage of collagenase positive cells ranged from 1 to 6% (early passage) to 35 to 46% (late passage) indicating that the late passage cells had higher basal levels of collagenase synthesis. Later passage cultures also secreted higher levels of immunoprecipitable collagenase into the culture medium and Northern analysis established that the basal level of collagenase mRNA was also 10 times greater in late passage cells. High basal levels of collagenase were also observed in fibroblasts cultured from an in vivo aged donor and from donors with Werner's syndrome. Collagenase production was induced in both early and late passage cell cultures by exposure to fibroblast extracellular matrix, fibroblast conditioned media, polypeptide growth factors, or phorbol esters. The induced levels were always greater in the late passage cell cultures than in the early passage cell cultures.

Blotting, Northern↗
PubMed1989

Chemical sterilization of bacterially contaminated bone without destruction of osteogenic potential.

A method to sterilize bone that has been contaminated or to assure sterility of allografts would be quite useful in the practice of orthopedics. The literature documents the osteogenic potential of demineralized bone matrix, prepared by treatment with hydrochloric acid and ethanol. We became interested in whether these potent antibacterial agents could render contaminated bone sterile and thus usable. Thus, we tested immersion in 70% ethanol or 0.6 N hydrochloric acid as sterilization methods for contaminated bone specimens. Bone fragments were (heavily) surface contaminated for 30-60 s by immersion in a suspension of either Escherichia coli, Staphylococcus aureus, or Bacillus subtilis and then placed in either the ethanol or hydrochloric acid solutions for 15 min, 30 min, 1 h, 2 h, 4 h, or 8 h. Subsequently, they were incubated in broth culture for 14 days. Using 70% ethanol, 80-90% of contaminated bone specimens were sterile in 4 h and all were sterile in 8 h. Using 0.6 N hydrochloric acid, only 10% of bone specimens contaminated with Staphylococcus aureus, 60% with Escherichia coli, and 90% with Bacillus subtilis were sterile in 8 h. Consequently, 70% ethanol could possibly by used as an antiseptic or sterilizing agent for contaminated bone, although we feel that such bone should also be demineralized. Ethanol should not be used for routine treatment of mineralized cortical bone as it has been shown to diminish the already poor osteogenic potential of cortical bone.

Administration, Topical↗
PubMed1989

[Effect of previous surgery on the results of total arthroplasty of the hip joint].

Clinical and radiological results as well/as complications after 123 total hip arthroplasties performed as secondary procedure have been evaluated. These patients underwent primary operation because of femoral neck fracture or intertrochanteric osteotomy in the treatment of hip osteoarthritis. The scores of clinical results according to the Merle d'Aubigené and Postel scale were as follows: pain 4.48, walk 3.99, movements 4.02. Radiological features of loosening of the cup were observed in 14.4 per cent and of the stem in 17.3 per cent of those investigated. Early infections were 3 times higher and summed with late infections constituted 8.8 per cent of those operated on. Deep infections and aseptic loosening caused revision in 11.5 per cent of those investigated.

Adult↗
PubMed1988

Serum and growth factors regulate expression of a 43 kDa protein in smooth muscle cell cultures.

Smooth muscle cells respond to injury and the presence of serum factors by modulating from a quiescent contractile cell to a motile synthetic phenotype. To evaluate the biochemical response to serum exposure, we examined the proteins synthesized and secreted in response to serum. The most prominent effect of serum was the rapid production of a protein with an apparent molecular weight of 43 kDa. Removal of serum from the culture environment led to a cessation of 43 kDa protein production. The effect of exogenous heparin on 43 kDa protein production was also evaluated. Neither the 43 kDa protein nor a previously described 38 kDa protein was induced by heparin. Further, heparin treatment did not counteract the effects of serum. These studies demonstrate that an early response of vascular smooth muscle cells to serum is the production of this previously undescribed protein and that other modifications of the culture conditions did not affect its synthesis.

Animals↗
PubMed1988

Differential response of early and late passage fibroblasts to collagenase stimulatory factor in conditioned media.

Comparison of the proteins secreted by early and late passage cultures of human fibroblasts reveals enhanced production of three proteins (Mr = 55,000, 58,000, and 61,000) in the late passage culture conditioned medium. The 55,000 and 58,000 Da proteins react with anticollagenase antibodies and are identified as procollagenase (Sottile, J. M. and Millis, A. J. T., J. Cell Biol. 106: 1518a, 1987). The production of immunoreactive collagenase was assayed using immunoprecipitation and immunofluorescence. Immunofluorescence microscopy revealed that each culture was heterogenous. The percentage of collagenase positive cells ranged from 1% (early passage) to 35% (late passage). Late passage cultures also secreted higher levels of immunoprecipitable collagenase into the culture medium than early passage cultures. High levels of collagenase production were also observed in fibroblasts cultured from donors with Werner's syndrome and from an aged donor. Media conditioned by the growth of human fibroblasts contained collagenase stimulating activity. In the presence of conditioned medium, the percentage of collagenase positive cells was increased to 10-20% (early passage) and 85% (late passage), indicating that each culture contains both responding and non-responding cells.

Antibodies, Monoclonal↗
PubMed1987

Enhanced synthesis of a Mr = 55,000 dalton peptide by senescent human fibroblasts.

The secretory protein profiles of early and late passage cultures of human fibroblasts were compared using polyacrylamide gel electrophoresis. In comparison with early passage cell cultures (40-50% lifespan completed), late passage (greater than 80% of lifespan completed) cell cultures exhibited enhanced production of several peptides in the Mr range 55-60,000. One of those peptides had an apparent molecular weight of Mr = 55,000 and was constitutively present in the late passage cell conditioned medium. Late passage cell cultures synthesized the Mr = 55,000 peptide in the presence or absence of fetal bovine serum. Serum did not enhance its production by early passage cells. Further, production of the peptide was not induced in early passage cell cultures whose proliferation was arrested either by serum starvation or by contact inhibition. Pulse chase studies demonstrated that the peptide appears in the culture medium within 60 min of labeling. There was no evidence that it is derived via degradation of other proteins present either in early passage or late passage cell conditioned media. Further, the production of the 55,000 dalton peptide did not appear to be regulated by factors present in conditioned media. The peptide was detected in the conditioned media produced by late passage cultures of several different cell strains.

Cell Survival↗
PubMed1986

In vitro expression of a 38,000 dalton heparin-binding glycoprotein by morphologically differentiated smooth muscle cells.

In vitro, high density monolayer cultures of vascular smooth muscle cells can be induced to form multicellular nodules. The nodular cells appear to be morphologically differentiated smooth muscle cells. Sodium dodecyl sulfate (NaDodSO4)-polyacrylamide gel electrophoresis was used to compare the proteins synthesized and secreted by monolayer and nodular cultures of smooth muscle cells. Although most proteins appeared to be similar, the nodular cultures contained a unique heparin binding protein of Mr = 38,000 (38kD protein) (Millis, A.J.T., Hoyle, M., Reich, E., and Mann, D.M., 1985, J. Biol. Chem., 260:3754-3761). The 38kD protein was glycosylated and its apparent molecular weight was shifted to Mr = 32,500 after synthesis in the presence of tunicamycin or digestion with endoglycosidase F. The production of 38kD protein by nodular cell cultures did not appear to result from the degradation of a high molecular weight precursor in nodular conditioned medium. Further, it was not detected in monolayer cell conditioned medium that had been incubated with nodular cells. Finally, its synthesis was not induced in monolayer cell cultures that had been labeled in nodular cell conditioned medium. The 38kD protein appears to be uniquely associated with nodular cultures of smooth muscle cells.

Animals↗
PubMed1985

Isolation and characterization of a Mr = 38,000 protein from differentiating smooth muscle cells.

In culture, vascular smooth muscle cells grow and form a confluent monolayer of cells. Under appropriate conditions, regions of the monolayer can be induced to draw away from the substrate and form multicellular nodules. The ultrastructure of the cells in the nodules appears to be similar to that of differentiated smooth muscle cells. The process of nodulation is associated with the synthesis of a unique protein whose molecular weight is estimated from gradient gel electrophoresis to be 38,000 (38-kDa Protein). The protein is secreted into the culture medium and can be detected either by metabolic labeling or by staining with Coomassie Blue. Partial purification of 38-kDa Protein was achieved using affinity chromatography. The protein is adsorbed to heparin-agarose, but not to gelatin-agarose. The concentration of 38-kDa Protein in nodular conditioned medium is estimated at 1.9 micrograms/ml and less than 0.01 microgram/ml in conditioned medium made from monolayer cells. The presence of 5% fetal bovine serum in the labeling medium does not affect 38-kDa Protein synthesis. Cross-reactivity with fibronectin was evaluated using polyvalent antibodies to 38-kDa Protein. The 38-kDa Protein is not antigenically related to fibronectin. Furthermore, we establish that the protein is not qualitatively influenced by the presence of ascorbate (50 micrograms/ml), beta-aminoproprionitrile fumarate (50 micrograms/ml) heparin (10 ng/ml), or fibronectin (20 micrograms/ml) in the culture medium. We find that the added components neither suppress 38-kDa Protein synthesis in nodular cultures nor enhance 38-kDa Protein synthesis in monolayer cultures. The 38-kDa Protein is not detected in either monolayer or nodular cell layers and appears to be a secreted protein. Its appearance in nodular conditioned medium during nodulation suggests a relationship with that process.

Animals↗
PubMed1985

Incorporation of cellular and plasma fibronectins into smooth muscle cell extracellular matrix in vitro.

Fibronectins isolated from the conditioned medium produced by cultures of undifferentiated (monolayer) and differentiated (nodular) swine vascular smooth muscle cells are similar but not identical. In general, the nodular-cell fibronectin has a smaller molecular mass than monolayer-cell fibronectin and appears to lack the COOH-terminal interchain disulfide linkage. We studied the incorporation of cellular and plasma fibronectins into the cell layer. Smooth muscle cells bound 2.5 times more monolayer-cell fibronectin than nodular-cell fibronectin. Polypeptide fragments of human plasma fibronectin were used as a model system to investigate fibronectin incorporation into the cell layer. Only intact molecules were incorporated into the cell layer and subsequently organized into fibers. Polypeptide fragments of molecular mass 205 kDa and 185 kDa were not incorporated even though they retained the collagen-, cell-, and heparin-binding regions. Incorporation appears to require an activity associated with either the NH2-terminal or COOH-terminal domains. We propose that fibronectin activity is lost during differentiation of smooth muscle cells.

Animals↗
PubMed1983

Preventable deaths in multiple trauma: review of deaths at Sunnybrook Medical Centre Trauma Unit.

One hundred and two patients, who have died of multiple injuries and on whom an autopsy was performed, have been reviewed. Injury severity was a major factor in the outcome and head injury played a definite role in more than half of the deaths. Missed diagnosis was not a major factor in the deaths. Future improvements will most likely be in the management of hemorrhage in patients with multiple injuries.

Adult↗
PubMed1981

Cell cycle dependent expression of a glucose regulated cell surface glycoprotein.

A cell surface associated "glucose regulated protein" has been described on nontransformed human fibroblasts. To examine the distribution of that protein on human fibroblasts specific antisera were used. The antisera was used in conjunction with indirect immunofluorescence and revealed that the glucose regulated protein was present as fibers on spread cells. Further, the antisera was used in complement mediated cytotoxicity assays to examine cells during specific stages of the mitotic cell cycle. Fibroblasts were synchronized by serum starvation, hydroxyurea inhibition, or colcemid inhibition followed by mitotic selection. The results demonstrated that the glucose regulated protein was maximally displayed during the G1 phase of the cell cycle and minimally displayed during the S and M phases.

Cell Line↗