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Biomedical subjects

M E Jones

Publications and source records attributed to M E Jones.

At least 145 records · Page 8Linked to original sources

Selective repression of actin and myosin heavy chain expression during the programmed death of insect skeletal muscle.

The intersegmental muscles (ISMs) of the tobacco hawkmoth Manduca sexta undergo two sequential changes in mass at the end of metamorphosis. Atrophy results in a 40% loss of muscle mass beginning 3 days before adult eclosion (emergence). Coincident with eclosion, the ISMs undergo programmed cell death that results in the complete destruction of the muscles during the subsequent 30 hr. These developmental changes are initiated by sequential decreases in the circulating titer of the ecdysteroids. While the molecular basis of ISM atrophy is largely unknown, the commitment of the muscles to die has been shown to require the repression of specific genes and the activation of others. Data presented here suggest that two of the repressed genes encode the proteins actin and myosin heavy chain. Expression of both actin and myosin heavy chain mRNA was greatly decreased when the ISMs became committed to die. When animals were treated with 20-hydroxyecdysone, cell death was delayed and the loss of these transcripts was reduced. At the protein level, actin expression was reduced by 84% at the time the muscles were committed to die. The reduction in actin and myosin heavy chain synthesis presumably plays a role in the rapid dissolution of the muscles.

Actins↗

Turnover of cholesterol and dipalmitoyl phosphatidylcholine in surfactant of adult rat lung.

In order to compare the turnover of two major surfactant components, [1 alpha,2 alpha (n)-3H]cholesterol and [methyl14C choline] dipalmitoyl phosphatidylcholine (DPPC) were introduced as lamellar bodies via the trachea into lightly anesthetized rats which were then allowed to recover. The radiotracers were assumed to have entered the alveolar surfactant pool and to have subsequently recycled in part into the lamellar bodies of alveolar type II cells. For DPPC, the specific activity vs. time curves of tubular myelin rich (alv-1) and tubular myelin poor (alv-2) alveolar lavage fractions were similar, and there was a plausible precursor-product relationship between lamellar bodies and either (or both) of these compartments. In contrast, however, the specific activities of alv-1 and alv-2 for cholesterol were quite different, allowing us to reject the hypothesis of a precursor-product relationship between classical lamellar bodies and alv-2. The estimated turnover time for DPPC in alv-1 was 240 or 206 min, depending on which subfraction of lamellar bodies one takes to be the precursor. For cholesterol it was 583 or 624 min. These longer turnover times for cholesterol should lead to a greater than twofold increase in the relative concentration of cholesterol in the putative product compartment. Such an increase was not found. We interpret this as reflecting either noncompartmental behavior of the alveolar surfactant pool, or multiple pools of lamellar bodies with different turnover times.(ABSTRACT TRUNCATED AT 250 WORDS)

1,2-Dipalmitoylphosphatidylcholine↗

Apert's Syndrome.

Explore the source record for details and available documents.

Acrocephalosyndactylia↗

Luria-Delbrück fluctuation analysis: estimating the Poisson parameter in a compound Poisson distribution.

Estimating the mutation rate from a Luria-Delbrück fluctuation experiment involves estimating the Poisson parameter in a compound Poisson distribution. The efficiency with which this can be estimated depends on how well the other random factors have been characterized. The assumption that cell growth can be represented as a stochastic pure birth or Yule process is biologically unrealistic but contributes little to the bias and variance of a maximum likelihood estimator of the mutation rate.

Algorithms↗

Sudden death in HIV-infected drug users. Presumptive overdose in an HIV-positive narcotic addict.

We report the case of an HIV-positive drug user who was found dead in a room in which were discovered tablets of dihydrocodeine and amitriptyline. There had been past episodes of drug overdose which was suspected of being the cause of death on this occasion. However toxicology analysis did not support this diagnosis and necropsy revealed a severe desquamative interstitial pneumonitis and HIV encephalitis. The need to investigate thoroughly the cause of sudden death in all HIV-positive drug users is emphasised.

Adult↗

Accounting for plating efficiency when estimating spontaneous mutation rates.

When the spontaneous mutation rate mu in mammalian cell cultures is estimated using Luria-Delbrück fluctuation analysis, many factors contribute to the unreliability of the estimate. Some of these have been documented by Featherstone et al. (1987) and by Kendal and Frost (1988). In particular, the plating efficiencies for mammalian cells are often much less than 100%, and this can be taken into account. A derivation of a generalized P0 estimator, mg, based on the classical P0 estimator of Luria and Delbrück (1943) is offered. In an experiment involving C cultures, of which z exhibit no mutant colonies, and in which the plating efficiency is p, (0 < p < 1), the estimated mean number of mutations per culture is given by [formula: see text] The classical P0 estimator is shown to be the limiting case of mg as plating efficiency tends to 100%.

Animals↗

An improved estimator of spontaneous mutation rates in Luria-Delbrück fluctuation experiments.

In estimating the spontaneous mutation rate, mu, in cultured mammalian cells, the number of mutant cells in several parallel cultures, each a clone from a single cell, is determined. Luria and Delbrück (1943), and subsequently Lea and Coulson (1949), proposed several estimators of mu using data from such experiments. These methods of analysis were originally proposed to apply to mutants arising in microbial cultures, and some of the limitations in applying them to mammalian cell cultures have been outlined by Featherstone et al. (1987), and by Kendal and Frost (1988). For a given value of mu, the number of mutants, r, found in a culture is exceedingly variable, so that r contains very little information concerning mu. It is important, therefore, that an estimator of mu be efficient; that it not waste the limited information available in the experimental data. To this end, a new estimator is proposed which compounds the information previously utilized by the so-called P0 and median estimators. Using a spreadsheet program such as that available with MINITAB, the algorithm can be programmed in as few as 26 instructions. Tested against simulated data across a range of values of mu, the proposed compound estimator is more efficient than the currently used estimators.

Algorithms↗

Altered expression of nerve growth factor in the skin of transgenic mice leads to changes in response to mechanical stimuli.

It has recently become clear that the neurotrophic factor, nerve growth factor, interacts specifically with nociceptive sensory neurons during development and maturity. Indeed, it may serve as a critical link between inflammation and the hyperalgesia that ensues in adult animals. Nerve growth factor is normally expressed in limiting amounts in target tissues of sensory and postganglionic sympathetic neurons. In the present study we have altered the basal level of nerve growth factor expression in the skin by producing transgenic mice that express a fusion gene construct containing either a sense or antisense nerve growth factor complementary DNA linked to the K14 keratin promoter. The K14-nerve growth factor transgene (sense or antisense) is abundantly expressed in skin from approximately embryonic day 15 and is then constitutively expressed throughout the life of the animal. In light of the fact that systemic administration of nerve growth factor to neonatal or adult rats leads to hyperalgesia, we have asked whether mice expressing the sense K14-nerve growth factor transgene exhibit similar sensory abnormalities and whether mice expressing the antisense nerve growth factor complementary DNA were hypoalgesic. Here we show that mice over-expressing nerve growth factor in skin display a profound hyperalgesia to noxious mechanical stimulation. Additionally, K14-nerve growth factor antisense mice displayed a profound hypoalgesia to the same stimuli.

Animals↗

Alcohol consumption and driving intentions amongst hotel patrons.

To examine the extent to which hotel patrons drink in excess of current health recommendations, and to identify risk factors for excessive drinking, hotel patrons were invited to participate in a survey of social drinking, which included a free breath test. Patrons were approached at 15 min. intervals, and 1000 subjects were studied. Amongst this group of hotel patrons interviewed, 1 in 2 had consumed alcohol in excess of the daily limit recommended by the National Health & Medical Research Council. One in 10 intending drivers had a BAC over the legal limit. Excessive drinking and drink driving appear to be prevalent amongst hotel patrons, and hotels should be targets for interventions designed to reduce these problems. For example, the National Guidelines for the Responsible Serving of Alcohol should be more widely practised.

Journal Article↗

An isoelastic prosthesis using a new composite material.

A new particulate composite material has been assessed with regard to the design of an 'isoelastic' or 'modulus matched' hip prosthesis. Three different prototype designs were assessed, each of which consisted of a femoral component made from the composite material, attached to a metal ball via a metal 'spike' insert. The prototypes varied in terms of the detailed shape of the spike, which was modified in the light of photoelastic stress analysis, so as to produce a more acceptable stress distribution to the composite material in the proximal region. Prototypes were made by hand moulding and by transfer moulding; both methods produced defects of various kinds. Simulation tests were conducted using a model of the proximal femur constructed from glass fibre composite, cyclically loaded in a servo-hydraulic testing machine. Though some difficulties were experienced with defective mouldings, especially for the transfer moulding process, a clear improvement was demonstrated for the final (Mark III) design. The fatigue endurance of this prototype was similar to that of conventional metal prostheses tested under similar conditions. Fatigue crack propagation tests were carried out on samples of the composite material to establish its propagation threshold. These results were combined with a finite element stress analysis and fracture mechanics theory to estimate the critical crack length for fatigue in this prosthesis. It was thus possible to specify the maximum safe size of defect that could be tolerated in use.

Biocompatible Materials↗

Blood recirculation in intravenous catheters for hemodialysis.

Long-term i.v. catheters for hemodialysis have the outflow tip extending approximately 2 to 3 cm beyond the inflow tip to prevent blood recirculation during dialysis; however, the lumens are frequently reversed because of inflow failure (i.e., inadequate flow when the inflow lumen is used for blood inflow into the dialyzer). Blood recirculation with reversed lumens (outflow lumen used for blood inflow) in inflow failure catheters and with standard and reversed lumens in well-functioning catheters was measured. Recirculation was measured at a blood flow of 300 mL/min. Systemic blood samples were taken after blood flow was slowed to 50 mL/min. Blood recirculation was calculated as a percentage of the difference between systemic and inflow lumen solute concentrations divided by the difference between systemic and outflow lumen solute concentrations. For each catheter, the recirculation values were calculated separately for urea and creatinine. Average recirculation as measured by both solutes was also calculated. Blood recirculations with standard lumens of well-functioning catheters, reversed lumens of well-functioning catheters, and reversed lumens of inflow failure catheters were 2.09 +/- 1.95, 13.58 +/- 9.87, and 7.10 +/- 5.12 (mean +/- SD), respectively. Whereas recirculation with standard lumens of well-functioning catheters is negligible, reversal of lumens causes considerable recirculation. Recirculation in inflow failure catheters with reversed lumens is significantly less than that with reversed lumens of well-functioning catheters. It was proposed that a blood clot attached at and/or immediately distal to the inflow lumen may disperse outflowing blood and diminish recirculation in inflow failure catheters.(ABSTRACT TRUNCATED AT 250 WORDS)

Blood Circulation↗

What school nurses really do--a study of school nurse utilization.

The functions of twelve school nurses in a large metropolitan school district were examined in relation to utilization of health services and student time lost from school using health problem categories. Findings suggest absentee data and utilization are valuable outcome measures supporting the importance of school health programming.

Absenteeism↗

Orotidylate decarboxylase: insights into the catalytic mechanism from substrate specificity studies.

Pyrimidine nucleotides were tested as substrates for pure yeast orotidylate decarboxylase in an attempt to gain insight into the nature of the catalytic mechanism of the enzyme. Substitutions of the 5-position in the pyrimidine ring of the orotidylate substrate resulted in compounds that are either excellent inhibitors or substrates of the enzyme. The 5-bromo- and 5-chloroorotidylates are potent inhibitors while the 5-fluoro derivative is a good substrate with a turnover number 30 times that observed with orotidylate. When carbon 5 of the pyrimidine ring is replaced by nitrogen in 5-azaorotidylate, the resulting compound is unstable in solution with a half-life of 25 min at pH 6. However, studies with freshly generated 5-azaorotidylate show that an enzyme-dependent reaction occurs, presumably decarboxylation. This enzyme reaction follows simple Michaelis-Menten kinetics. Because the 5-aza group is not electrophilic, an enzyme mechanism utilizing a nucleophilic addition of the enzyme at the 5-position is ruled out. We also present studies that are not compatible with a mechanism requiring the formation of a Schiff's base prior to decarboxylation. The enzyme is tolerant of modest substitution at the 4-position, for the 4-keto group can be replaced with a thioketone. However, no catalysis is observed when the same substitution is made at the 2-position. Similarities in the substrate specificity of orotate phosphoribosyltransferase and orotidylate decarboxylase led us to compare the amino acid sequences of the two enzymes; significant (20%) sequence homology was observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗

A unique catalytic and inhibitor-binding role for Lys93 of yeast orotidylate decarboxylase.

The presence of a proton-donating catalytic amino acid side chain in orotidylate decarboxylase (ODCase) was sought by site-directed mutagenesis. Replacement of yeast ODCase Lys93 with a cysteine resulted in a mutant protein (K93C) with no measurable activity, representing a decrease in activity by a factor of, at most, 2 x 10(-8) times the activity of the wild-type enzyme. Treatment of this mutant protein with 2-bromoethylamine, designed to append Cys93 to yield S-(2-aminoethyl)cysteine, restored activity by a factor of at least 5 x 10(5) over the untreated mutant protein. Activity could not be restored by treatment with other brominated reagents designed to replace the epsilon-amino of S-(2-aminoethyl)Cys93 with a different functional group. The overall architecture of the K93C protein was not significantly changed, as judged by the similar dimerization properties (in the absence of ligands) of the mutant enzyme compared to the wild-type enzyme. The binding affinity of the substrate orotidylate was not measurably changed by the mutation, indicating that Lys93 has an essential role in catalysis which is mechanistically distinguishable from substrate binding. Apparently the mutation removes an integral portion of the active site and does not drastically affect the structural or substrate binding properties. However, the affinities of the mutant protein for the competitive inhibitors 6-azauridylate (6-azaUMP) and UMP are significantly altered from the pattern seen with the wild-type enzyme. The K93C protein has an affinity for the neutral ligand UMP which is greater than that for the anionic 6-azaUMP, in clear contrast to the preference for 6-azaUMP displayed by the wild-type enzyme. Lys93 is apparently critical for catalysis of the substrate to product and for the binding of anionic inhibitors; the data are discussed in terms of previously existing models for transition-state analogue inhibitor binding and catalysis.

Animals↗

Analysis of algebraic weighted least-squares estimators for enzyme parameters.

An algorithm for the least-squares estimation of enzyme parameters Km and Vmax. is proposed and its performance analysed. The problem is non-linear, but the algorithm is algebraic and does not require initial parameter estimates. On a spreadsheet program such as MINITAB, it may be coded in as few as ten instructions. The algorithm derives an intermediate estimate of Km and Vmax. appropriate to data with a constant coefficient of variation and then applies a single reweighting. Its performance using simulated data with a variety of error structures is compared with that of the classical reciprocal transforms and to both appropriately and inappropriately weighted direct least-squares estimators. Three approaches to estimating the standard errors of the parameter estimates are discussed, and one suitable for spreadsheet implementation is illustrated.

Algorithms↗

Interstate differences in incidence and mortality from melanoma. A re-examination of the latitudinal gradient.

OBJECTIVE: To investigate the patterns of cutaneous malignant melanoma (CMM) mortality in Australia. DESIGN: A descriptive analysis of melanoma incidence and mortality in Australia supplemented by a case series analysis of melanoma survival. Melanoma mortality rates were based on tabulations supplied by the Australian Bureau of Statistics for the years 1969-1989. Melanoma incidence rates were based on State cancer registry records for the years 1977-1990. The case series survival analysis was based on detailed individual records from the population-based cancer registries in Tasmania and South Australia. MAIN OUTCOME MEASURES: The level of and rise in melanoma mortality rates during 1969-1989 in Australia; the five-year survival rates for Tasmanian and South Australian cases; and male:female incidence ratios related to latitude. RESULTS: We found annual increases in melanoma mortality rates of 2.5% in men (P < 0.0001) and 1.1% in women (P < 0.0001) for all Australia. The five-year survival rates (with 95% confidence intervals [CI]) were: 67% (59%-75%) for Tasmanian men; 79% (76%-83%) for South Australian men; 80% (74%-86%) for Tasmanian women and 88% (86%-91%) for South Australian women. A change in the male:female incidence ratio with latitude was also found--women have significantly higher incidence rates at higher latitudes, but similar rates to men at lower latitudes. CONCLUSIONS: The age standardised mortality from CMM for the period 1969 to 1989 shows little variation by State for women, despite a considerable range in latitude. CMM mortality in men is increasing at a faster rate than that in women. Between 1982 and 1987 the male:female incidence ratio in high latitudes in the Southern Hemisphere showed an excess of cases in women, a finding which we believe has not been reported before.

Aged↗

Inhibition kinetics of hepatic microsomal long chain fatty acid-CoA ligase by 2-arylpropionic acid non-steroidal anti-inflammatory drugs.

Microsomal long chain fatty acid CoA ligase (EC 6.2.1.3) has been implicated in the formation of CoA thioesters of xenobiotics containing a carboxylic acid moiety. In this study we have demonstrated that the microsomal enzyme from rat liver exhibits biphasic kinetics for the formation of palmitoyl-CoA, i.e. there are high affinity low capacity Kmhigh, 1.6 microM, Vmaxhigh, 12.9 nmol/mg/min) and low affinity high capacity (Kmlow, 506 microM, Vmaxlow, 58.3 nmol/mg/min) components. Inhibition of the high affinity isoform was studied using the R and S enantiomers of ibuprofen, fenoprofen, ketoprofen and naproxen. The high affinity component of palmitoyl-CoA formation was competitively inhibited by R-fenoprofen (Ki 15.4 microM) while R-ibuprofen exhibited mixed inhibition kinetics. In contrast the R and S enantiomers of ketoprofen and naproxen were non-competitive inhibitors. This diversity of inhibition kinetics observed argues in favour of a binding site in addition to the catalytic site. A competitive interaction with the high affinity form correlated with literature evidence of enantiospecific chiral inversion and "hybrid" triglyceride formation for the R enantiomers of fenoprofen and ibuprofen. Paradoxically, R-ketoprofen which is extensively inverted in rats was a non-competitive inhibitor of palmitoyl-CoA formation by the high affinity isoform suggesting that it may not act as an alternate substrate. The results of this study clearly indicate that formation of R-2-arylpropionate-CoAs is not fully explained by interaction with the high affinity isoform of a microsomal long chain (palmitoyl) CoA ligase and therefore the involvement of other isoforms cannot be discounted.

Animals↗