PubMed Health⌕ Search

Biomedical subjects

M L Taylor

Publications and source records attributed to M L Taylor.

At least 19 recordsLinked to original sources
PubMed1992

Interaction of androgen and 1,25-dihydroxyvitamin D3: effects on normal rat bone cells.

We studied the actions of testosterone (T) and 5 alpha-dihydrotestosterone (DHT) in combination with 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] on primary rat bone cells. The actions of androgens were generally anabolic, although response patterns varied considerably between cultures. For example, DHT caused striking dose- and time-dependent increases in [3H]thymidine incorporation into calvarial cells over the range 1-100 nM, with maximal stimulation of 2.5-fold after 9 days in culture. Testosterone (50 nM) also stimulated [3H]thymidine incorporation into long bone-derived cells. 1,25-(OH)2D3 generally blunted or abolished the proliferative action of androgens but was not itself always inhibitory; in some experiments, stimulation of [3H]thymidine incorporation occurred. Collagen production, as assessed by [3H]proline incorporation into pepsin-resistant protein secreted by calvarial cells, was also stimulated by DHT. In some cultures, androgen responses were absent, although striking inhibitory responses to 1,25-(OH)2D3 were observed. These results illustrate complex patterns of responses to androgens and 1,25-(OH)2D3 in cells derived from rat bone.

Alkaline Phosphatase↗
PubMed1992

Effect of retinoic acid on the resorptive activity of chick osteoclasts in vitro.

Mixed cell suspensions mechanically isolated from the long-bones of day 20 prehatch chicks were cultured for 24 h on bone and sperm whale dentine slices in the presence of 0, 10, 100, and 1000 nM retinoic acid (RA). Significant inhibitions in the numbers of discrete lacunae resorbed per dentine slice, and in the ratio of lacunae per tartrate-resistant acid phosphatase-positive multinuclear cell were observed with all concentrations of RA studied. Semi-automated, 3-dimensional analysis of 733 pits was performed on one series of experiments using a tandem scanning (confocal) microscope, interfaced to an image analyzing computer. The majority of lacunae were small and unilocular; the plan areas of 90% of control pits were below 500 microns 2. Small but statistically significant increases in lacunar areas, but not mean or maximum depths or volumes, were observed in the presence of 10 and 100 nM RA; however, these changes were much smaller than the magnitude of the decrease in pit numbers. Thus, the overall effect of RA in this system was inhibitory. Our findings contrast with the well known stimulatory action of retinoids on bone resorption both in vivo and in organ culture, but may parallel the inhibitory effects of prostaglandins observed in disaggregated osteoclast systems.

Animals↗
PubMed1992

The effects of varying doses of aspirin on human platelet activation induced by PAF, collagen and arachidonic acid.

1. The effect of increasing doses of orally administered aspirin (30-900 mg) on platelet aggregation and ATP release induced by arachidonic acid (AA), collagen and platelet activating factor (PAF) was assessed in 12 normal volunteers. 2. Aspirin ingestion was associated with a significant increase in EC50 for AA (P less than 0.0001) and collagen (P less than 0.0001) but not for PAF (P greater than 0.495) although the normal biphasic aggregation response for the latter was abolished. Maximum ATP release was reduced by aspirin for all three agonists. 3. The mean maximum degrees of inhibition of platelet aggregation induced by aspirin for AA, collagen and PAF were 100%, 48% and 21% of baseline, respectively. The corresponding mean maximum inhibition of ATP release was 100%, 63% and 57%. The minimum cumulative doses of aspirin producing these effects were 240, 240 and 90 mg for AA, collagen and PAF respectively. For collagen alone, there was a significant decrease in the degree of inhibition of aggregation between the last dose on day 1 (150 mg) and the baseline measurement on day 2. 4. Platelets from female subjects were more sensitive to collagen (P less than 0.05) and AA (P less than 0.01) stimulation compared with males. However, prior to aspirin ingestion, PAF produced a greater maximum response in platelets from females (P less than 0.02) while following aspirin ingestion PAF-induced activation was inhibited to a greater degree in females (P less than 0.02). 5. These results indicate that collagen- and PAF-induced platelet activation are only partially dependent on cyclo-oxygenase and for PAF this seems related only to the second phase of aggregation.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗
PubMed1992

5-Nitro-4-(N,N-dimethylaminopropylamino)quinoline (5-nitraquine), a new DNA-affinic hypoxic cell radiosensitizer and bioreductive agent: comparison with nitracrine.

Targeting of electron-affinic radiosensitizers to DNA via noncovalent binding (e.g., intercalation) may offer the potential for increasing sensitizing efficiency. However, it has been suggested that high-affinity DNA binding may compromise sensitization by restricting the mobility of sensitizers along the DNA, and by decreasing rates of extravascular diffusion in tumors. The weak DNA intercalator nitracrine (1-NC) is a more efficient radiosensitizer than related nitroacridines with higher DNA-binding affinities (Roberts et al., Radiat. Res. 123, 153-164, 1990). The present study investigates whether electron-affinic agents of even lower DNA-binding affinity may be superior to nitroacridines. The quinoline analog of 1-NC, 5-nitraquine (5-NO), was shown to have an intrinsic association constant for calf thymus DNA in 20 mM phosphate buffer which was 12-fold lower than that of 1-NC. 5-Nitraquine was not accumulated as efficiently as 1-NC by AA8 cells, but, despite a similar one-electron reduction potential, was 2- to 3-fold more potent than 1-NC as a hypoxia-selective radiosensitizer in vitro when compared on the basis of average intracellular concentration. Thus the radiosensitizing potency of 5-NQ appears not to be compromised by its low DNA-binding affinity. The cytotoxic mechanisms of 5-NQ and 1-NC appear to be similar (hypoxia-selective formation of DNA monoadducts), but 5-NQ is 1200-fold less potent than 1-NC as a cytotoxin. Despite this advantage, 5-NQ was not active in vivo as a radiosensitizer in SCCVII tumors. This lack of activity appears to be due to its relatively high toxicity in vivo (intraperitoneal LD50 of 105 mumol kg-1 in C3H/HeN mice), high one-electron reduction potential (-286 mV), and rapid metabolism to the corresponding amine in mice. The in vitro therapeutic index (hypoxic radiosensitizing potency/aerobic cytotoxic potency) of this weak DNA binder was lower than that of the non-DNA targeted radiosensitizer misonidazole, suggesting that DNA targeting enhances cytotoxicity more than radiosensitization. Development of useful DNA-targeted radiosensitizers may require the exploitation of DNA binding modes different from those of the nitroacridines and nitroquinolines.

Aminoquinolines↗
PubMed1991

Arg-Gly-Asp (RGD) peptides and the anti-vitronectin receptor antibody 23C6 inhibit dentine resorption and cell spreading by osteoclasts.

Studies with a range of monoclonal and polyclonal antisera to components of the human, rat, and chick vitronectin receptor, alpha V beta 3, and the VLA beta 1 chain show that chick and rat osteoclasts express similar integrin receptors to those described in man. Biochemical analysis with monoclonal antibody 23C6 confirmed the presence on chick osteoclasts of a vitronectin receptor heterodimer of similar size (110/95 kDa reduced) to that immunoprecipitated from human osteoclastoma giant cells. The synthetic peptide GRGDSP, corresponding to the cell adhesion sequence in fibronectin, but not GRGESP peptide, induced significant (P less than 0.005) osteoclast retraction in chick and rat osteoclasts at IC50s (+/- SEM) of 210.0 +/- 14.4 and 191.4 +/- 13.7 microM, respectively; monoclonal anti-vitronectin receptor alpha V beta 3 complex antibody, 23C6, produced similar changes in chick osteoclasts (IC50 = 1.45 +/- 0.22 microM). Antibody 23C6 inhibited the number of pits resorbed in dentine by chick osteoclasts over a concentration range of 4.4 to 88 micrograms/ml; a significant 76% reduction (P = 0.03) was observed at a final concentration of 88 micrograms/ml (6 microM). The effect of peptides upon dentine resorption was less dramatic. No consistent inhibition was seen using chick osteoclasts. Inhibitory effects on resorption by rat osteoclasts were, however, observed; significant reduction in resorption occurred with both GRGDSP (78%; P less than 0.01) and GRGESP (67%; P = 0.02) peptides at 400 microM peptide concentration. These data demonstrate that osteoclast function can be disrupted by low concentrations of the anti-vitronectin receptor antibody, 23C6. The inhibitory effects of the peptides used in this study produced effects on dentine resorption which were generally weaker and variable, although osteoclast cell adhesion was consistently inhibited in an Arg-Gly-Asp (RGD)-dependent manner. We conclude that the vitronectin receptor may play an important role in effecting resorption of mineralized tissues by osteoclasts.

Amino Acid Sequence↗
PubMed1991

Demonstration that phosphorescent 6-bromo-2-naphthyl sulfate can be used to probe heme accessibility in heme proteins.

The phosphorescence properties of 6-bromo-2-naphthyl sulfate (BNS) in aqueous solution were studied. The phosphorescence lifetime is several hundred microseconds and is self-quenched. Although a fluorescent photoproduct is formed from BNS, it does not interfere with the decay properties of triplet-state BNS and its utility as a probe of the accessibility of the heme group in heme proteins. Quenching of BNS phosphorescence does not occur for the non-heme protein lysozyme and apomyoglobin but occurs by a dynamic mechanism with a quenching constant of 1-2 x 10(9) M-1 s-1 for cytochrome c and myoglobin and with a quenching constant of 6.2 x 10(9) M-1 s-1 for protoporphyrin IX. The phosphorescence of an inclusion complex of 1-bromonaphthalene and beta-cyclodextrin is not quenched by heme-containing proteins. The temperature and viscosity dependencies of the rate with which BNS phosphorescence is quenched by microperoxidase-11 are consistent with unit quenching efficiency. These results indicate that quenching of BNS phosphorescence occurs only upon contact with the quencher, and the quenching constant can be used to assess the degree of accessibility of the heme group.

Fluorescent Dyes↗
PubMed1990

The effect of fluoride on the resorption of dentine by osteoclasts in vitro.

Osteoclasts were isolated mechanically from bones of prehatch chicks and cultured on dentine for 24 or 39 h in medium without or with added sodium fluoride at concentrations of 0.15, 1, 15, or 30 mg/liter. Fixed and toluidine blue-stained specimens were examined by light microscopy and osteoclasts were counted, all cells were removed and resorption pits were counted. The dentine was then prepared for scanning electron microscopy (SEM) and pits located on a raster search using backscattered electron imaging. Stereophotogrammetric measurements were made of the depths, plan and true surface areas and volumes of pits, and the volume:area ratio and a three-dimensional form factor calculated for each. The presence or absence of a ring of demineralization around each pit was recorded. Pits with the same plan area were significantly (p less than 0.001) less deep in cultures containing 15 or 30 mg/liter NaF and the incidence of rings around pits was higher (controls 2.7%, 15 mg/liter 31.3%, 30 mg/liter NaF 39.7%). In one experiment, pits were larger in the specimen cultured with added 1 mg/liter NaF, but in a much larger series of 631 (251 in the control and 380 in the 1 mg/liter NaF cultures) pits measured, depths, volumes and volume:plan-area ratios were significantly lower (p less than 0.001), and rings round pits 2.5 times more common, in the fluoride-treated group. A complex interplay of stimulation of some cells in the culture with indirect stimulation of osteoclasts and direct inhibition of osteoclastic resorptive function may be operating at this concentration of fluoride. In vivo, therapeutic doses of sodium fluoride would be expected to act not only to increase the formation of osteoid but also to suppress osteoclastic activity.

Animals↗
PubMed1990

Nutritional status and growth in juvenile rheumatoid arthritis.

The specific cause of short stature in juvenile rheumatoid arthritis (JRA) is unknown. One hypothesis links altered growth to inadequate dietary intake. In this study, nutritional status was assessed in 34 children with JRA (8 with systemic JRA, 14 with polyarticular JRA, and 12 with pauciarticular JRA) and 9 healthy controls using 3-day diet records, anthropometrics, and biochemical analyses. Differences in growth were found among the three types of JRA. One third of all subjects were at or below the 10th percentile in height for age (these being predominantly among the systemic and polyarticular groups). With few exceptions, the mean dietary intake for calories and essential nutrients was found to be adequate for each of the three groups. However, more than half of those with systemic JRA reportedly consumed less than the recommended caloric intake for their age and weight. No significant correlations were found linking dietary intake to growth percentiles in any of the groups studied. Biochemical abnormalities were found among the systemic and polyarticular groups. These abnormalities included low plasma levels of vitamins A and C, proteins (albumin, prealbumin, and retinol binding protein) and zinc; and increased levels of copper and glutathione peroxidase activity. Plasma selenium and vitamin E levels were unchanged. The discrepancy between intake and certain circulating nutrient levels may reflect alterations in the requirements, absorption, or use of these nutrients in the presence of chronic inflammation.

Adolescent↗
PubMed1990

Fluorescence in the tandem scanning microscope.

Our studies have shown that the fluorescence mode can be used to good effect in both tandem scanning microscopes (TSM: direct view confocal microscopes) as well as confocal scanning laser microscopes (CSLM). Applications are presented which show that the two great advantages of TSM are real-time viewing and real colour, which allow faster use and interpretation. CSLM are complementary, not competitive, being currently more sophisticated for low-level fluorescence work. This is equally possible with available TSM, but requires further development using CCD cameras and image-processing systems.

Animals↗
PubMed1990

Confocal fluorescence microscopy: some applications in bone cell biology.

Three-dimensional information is necessary for the proper investigation of the interrelationships of bone cells, and of the complex interface between these cells and the bone matrix they form and destroy. The use of fluorescence confocal microscopy was explored in the determination of the distribution of immunolabelled actin and vinculin--cytoskeletal and attachment proteins--in isolated chick bone cells cultured on dentine, and in neonate rat and rabbit calvaria. Confocal imaging, compared with conventional fluorescence imaging, greatly enhanced the interpretation possible.

Actins↗
PubMed1990

Tuberculous anergic sera or purified protein derivative treatment induces modification in lymphocyte transformation of cells from patients with tuberculosis.

Peripheral blood mononuclear cells from 20 purified protein derivative (PPD)-reactive (PPD+) tuberculous patients were cultured in autologous, tuberculous anergic, or normal serum. After 12 h of incubation, the serum was eliminated and lymphocyte transformation with PPD was performed. Transformation was inhibited only in cells incubated with anergic serum. In contrast, cells from 11 anergic tuberculous (PPD-) individuals recovered the ability to respond to an optimal PPD dose after treatment with high PPD concentrations followed by several washings. The cells which recovered returned to their initial anergic state when incubated with sera from anergic patients. Under both conditions, incubation with sera did not abolish the response to the mitogen phytohemagglutinin. Cells from healthy PPD+ or PPD- individuals were used as controls. The most important finding derived from serum analysis was the increased levels of specific immunoglobulins G and A in anergic patients.

Adolescent↗
PubMed1989

The effect of fluoride on the patterns of adherence of osteoclasts cultured on and resorbing dentine: a 3-D assessment of vinculin-labelled cells using confocal optical microscopy.

Osteoclasts were isolated mechanically from chick long bones and cultured on dentine for three days in MEM/10% FCS with or without 1 mg/l NaF, (near to therapeutic ranges for serum fluoride levels) or 15 mg/l NaF (which has been found to reduce the volume:plan-area ratios of resorption pits in vitro). The distribution of close contacts with the substratum was determined by immunolabelling of vinculin and confocal fluorescence microscopy. Four characteristic patterns of contacts were observed: infilled discs, rings, crescents and patches: these reflect the progress of the cell in the process of pit formation. Cell-substratum adherence was more extensive than previously reported with contacts often extending down the sides of pits and sometimes across the floor. This distribution explains the curved shape of unilocular pits and the kidney-shaped extensions of multilocular pits. The peripheral creeping of the contact region was more marked in the osteoclasts of the 15 mg/l NaF cultures, and these cultures showed a decrease in pit depth and inhibition of osteoclastic movement in the presence of fluoride.

Animals↗
PubMed1989

A note on susceptibility of Branhamella catarrhalis to heavy metals.

The susceptibility of 56 strains of Branhamella catarrhalis and ten Neisseria spp. to arsenate, silver, nickel, mercury, lead, cadmium, chromium, manganese, iron, cobalt and molybdenum was tested with an agar dilution technique. All but two strains of B. catarrhalis were resistant to multiple metal ions. There were not sufficient differences in susceptibility, however, to allow the development of a typing scheme based on resistograms. Heavy metal resistance in Branhamella was unrelated to beta-lactamase production. Neisseria spp. were more susceptible to metal ions than B. catarrhalis and this may form the basis of a simple diagnostic test.

Drug Resistance, Microbial↗
PubMed1989

Characterization of immunologically important antigens and allergens of Aspergillus fumigatus.

Using a variety of immunochemical methods, including quantitative immuno-electrophoretic techniques, combined with gel filtration and iso-electric focusing, and production of monospecific antisera for identification and affinity purification, 4 major components of Aspergillus fumigatus have now been partially characterized. Numbering of these was derived from a reference allergic bronchopulmonary aspergillosis (ABPA) self-crossed radio-immuno-electrophoresis pattern of reactivity. Two major intracellular/cytoplasmic, concanavalin A (Con A)-binding antigens, Ag 7 and Ag 13, of molecular weights 150-200 and 70 kilodaltons (kD), respectively, were confirmed to be of importance for both ABPA and aspergilloma in specific sandwich enzyme-linked immunosorbent assays. A rapidly released component, Ag 5, of molecular weight 35 kD, proved both antigenic and allergenic, with aspergilloma patients having especially high-titre IgG antibodies. The major allergenic component Ag 3, of molecular weight 24 kD by gel filtration and 18 kD by SDS-PAGE was, like Ag 5, relatively heat-labile and non-Con-A-binding. Interestingly, T cell clones have been identified which respond primarily to an 18-kD fraction.

Allergens↗
PubMed1989

Relationship between uterine secretion of prostaglandin F2 alpha induced by oxytocin and endogenous concentrations of estradiol and progesterone at three stages of the bovine estrous cycle.

The purpose of this experiment was to determine whether differences among cows in the ability of oxytocin to stimulate uterine secretion of prostaglandin F2 alpha (PGF2 alpha) were related to the endogenous ovarian steroid environment. Sexually mature heifers were treated with oxytocin (.33 IU/kg BW) at three stages of the estrous cycle: early (d 3 to 5; n = 5), middle (d 10 to 11; n = 5) or late (d 16 to 17; n = 5). To assess uterine responsiveness to oxytocin, concentrations of 13,14-dihydro-15-keto-PGF2 alpha (PGFM) were quantified in jugular venous plasma samples collected at 1/2-h intervals for 8 h postinjection. The ovarian steroid environment at the time of injection was estimated by measuring the concentrations of progesterone and estradiol in jugular venous plasma samples collected at 4-h intervals for 12 h immediately prior to injection. Concentrations of PGFM increased immediately following injection of oxytocin either early or late in the estrous cycle. The response was much less during the middle of the estrous cycle. The magnitudes of response, early and late in the estrous cycle, were similar and greater than that observed during the middle of the estrous cycle (P less than .05). There was a positive relationship (R2 greater than .8; P less than .05) between magnitude of the response to oxytocin and ratio of estradiol to progesterone both early and late in the estrous cycle. Thus, individual differences in uterine secretion of PGF2 alpha in response to oxytocin were related to stage of the cycle and to differences in the endogenous ovarian steroid environment within each stage of the estrous cycle.

Animals↗