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N Craig

Publications and source records attributed to N Craig.

At least 19 recordsLinked to original sources
PubMed1995

The limits of the possible: models of power supply and demand in cycling.

This paper outlines a general strategy for mathematical modeling of cycling performance. This strategy involves formulating one expression describing the power available for external work from physiological sources. The variables used in this expression include maximal aerobic power (VO2max), fractional utilisation of VO2max, mechanical efficiency, maximal accumulated oxygen deficit, and the time constants relating to the expression of aerobic and anaerobic capacities. A second expression describing the power demand of cycling is then constructed. The variables used in this expression include the mass, projected frontal area and drag characteristics of the system, the coefficient of rolling resistance, environmental variables such as temperature, barometric pressure, relative humidity, wind speed and direction and the slope of the course. The two expressions are equated and solved using an iterative procedure. Two series of trials were used to assess the predictive accuracy of the model, one using track endurance performances and the other a 26 km road time-trial. The correlations between actual and predicted times have been excellent (0.92-0.95, p < or = 0.0001), with small mean differences (0-1.83% of mean performance time) and mean absolute differences (1.07-3.24%). The model allows us to make predictions about the effect of equipment changes and environmental factors, to compare performances under very different conditions, and to predict the limits of the possible in cycling performance. A range of options designed to improve cycling performance is described.

Bicycling↗
PubMed1994

Lanterns in the jungle: is the NHS driven by the wrong kind of efficiency?

The performance of the National Health Service is assessed in part by an Efficiency Index (EI) which is applied to the service as a whole as well as to individual health authorities. The EI relates increases in the amount of patient care activity to increases in total expenditure. The index can give a misleading impression of performance, creates perverse incentives and is at odds with the overall strategy of the health service which is to place greater emphasis on the promotion of health and to provide more care in primary and community-based settings. The philosophy, validity and appropriateness of the EI are discussed.

Community Health Services↗
PubMed1994

Processing of eukaryotic ribosomal RNA.

In summary, it can be argued that the understanding of eukaryotic rRNA processing is no less important than the understanding of mRNA maturation, since the capacity of a cell to carry out protein synthesis is controlled, in part, by the abundance of ribosomes. Processing of pre-rRNA is highly regulated, involving many cellular components acting either alone or as part of a complex. Some of these components are directly involved in the modification and cleavage of the precursor rRNA, while others direct the packaging of the rRNA into ribosome subunits. As is the case for pre-mRNA processing, snoRNPs are clearly involved in eukaryotic rRNA processing, and have been proposed to assemble with other proteins into at least one complex called a "processosome" (17), which carries out the ordered processing of the pre-rRNA and its assembly into ribosomes. The formation of a processing complex clearly makes possible the regulation required to coordinate the abundance of ribosomes with the physiological and developmental changes of a cell. It may be that eukaryotic rRNA processing is even more complex than pre-mRNA maturation, since pre-rRNA undergoes extensive nucleotide modification and is assembled into a complex structure called the ribosome. Undoubtedly, features of the eukaryotic rRNA-processing pathway have been conserved evolutionarily, and the genetic approach available in yeast research (6) should provide considerable knowledge that will be useful for other investigators working with higher eukaryotic systems. Interestingly, it was originally hoped that the extensive work and understanding of bacterial ribosome formation would provide a useful paradigm for the process in eukaryotes. However, although general features of ribosome structure and function are highly conserved between bacterial and eukaryotic systems, the basic strategy in ribosome biogenesis seems to be, for the most part, distinctly different. Thus, the detailed molecular mechanisms for rRNA processing in each kingdom will have to be independently deciphered in order to elucidate the features and regulation of this important process for cell survival.

Animals↗
PubMed1993

Preferential transposition of Drosophila P elements to nearby chromosomal sites.

Two different schemes were used to demonstrate that Drosophila P elements preferentially transpose into genomic regions close to their starting sites. A starting element with weak rosy+ marker gene expression was mobilized from its location in the subtelomeric region of the 1,300-kb Dp1187 minichromosome. Among progeny lines with altered rosy+ expression, a much higher than expected frequency contained new insertions on Dp1187. Terminal deficiencies were also recovered frequently. In a second screen, a rosy(+)-marked element causing a lethal mutation of the cactus gene was mobilized in male and female germlines, and viable revertant chromosomes were recovered that still contained a rosy+ gene due to an intrachromosomal transposition. New transpositions recovered using both methods were mapped between 0 and 128 kb from the starting site. Our results suggested that some mechanism elevates the frequency 43-67-fold with which a P element inserts near its starting site. Local transposition is likely to be useful for enhancing the rate of insertional mutation within predetermined regions of the genome.

Animals↗
PubMed1991

Sequence organization and RNA structural motifs directing the mouse primary rRNA-processing event.

The first processing step in the maturation of mouse precursor rRNA involves cleavage at nucleotide ca. +650, at the 5' border of a 200-nucleotide region that is conserved across mammals and contains the sequences that direct the processing. To identify the relevant sequence elements, we used rRNAs with small internal mutations and short pre-rRNA substrates. Much of the region can be mutated without appreciable effect, but nucleotides +655 to +666 appear to be absolutely required and short segments surrounding +750 and +810 markedly stimulate processing. The minimal processing signal corresponds to rRNA nucleotides +645 to +672. Formation of a ribonucleoprotein complex of retarded electrophoretic mobility is evidently necessary but not sufficient for processing. Computer-assisted analysis suggested a phylogenetic- and mutant-supported secondary structure in which the minimal processing signal forms a stem with the +655 region in the loop, and there is a separate branched duplex containing the downstream stimulatory sequences. Use of antisense RNA, in trans and in cis, to sequester the +655 region in a duplex supported the hypothesis that this critical region was needed in a single-stranded conformation for processing and for specific complex formation.

Animals↗
PubMed1988

Umbilical cord knots and encirclements.

Although cord knots and/or encirclements account for 1 in 10 stillbirths of infants weighing 2,500 g or more, no problem due to this cause was encountered in a prospective study of 1,115 vaginal deliveries. In this study there were 6 cases of cord knot (0.5%) and 158 of cord encirclement (14.2%). The range of cord length was 27-122 cm, the 10th, 50th and 90th percentiles being 40, 52 and 69 cm respectively. In this study there was no clinical warning (fetal distress) of cord encirclement or knot during pregnancy, labour or delivery.

Asphyxia Neonatorum↗
PubMed1987

Nucleotide sequence determining the first cleavage site in the processing of mouse precursor rRNA.

The first step in the processing of 47S precursor rRNA in mouse cells is reproduced in vitro in an S-100 transcription reaction and consists of an endonucleolytic cleavage at residue +650 of the primary transcript followed by rapid degradation of the fragment upstream from residue +650. An analogous processing occurs in human rRNA. The mouse and human rRNA sequences are approximately equal to 80% conserved for 200 nucleotides on the 3' side of these processing sites, suggesting that this conserved region may be important in specifying the processing. To test this hypothesis, we constructed a systematic series of deletion mutants approaching the mouse rDNA processing region from both the 5' and 3' directions and analyzed the processing of their transcripts in vitro. The 5' boundary of the region required for processing is quite sharp and corresponds to the rRNA cleavage site at the 5' end of the conserved sequence region. The 3' boundary is more complex: The 3' deletions extending to between 250 and 130 nucleotides beyond the processing site cause about a 50% decrease in the amount of the processed RNA. A 3' deletion that extends to 109 nucleotides beyond the processing site greatly reduces the processing efficiency. Deletions to or beyond 91 nucleotides on the 3' side of the processing site virtually eliminate processing. Under altered ionic conditions, transcripts of 3' deletions extending to only 41 nucleotides beyond the processing site can still direct a low level of accurate processing. These results demonstrate that the mouse/human conserved sequence just on the 3' side of the primary rRNA processing site consists of several domains that direct and/or augment both the initial endonucleolytic cleavage and the closely coupled selective degradation of the upstream fragment that together constitute the primary rRNA processing event.

Animals↗
PubMed1987

Primary processing of mammalian rRNA involves two adjacent cleavages and is not species specific.

The primary transcript of the mouse rRNA gene is rapidly processed at nucleotide approximately +650 both in vivo and in vitro. Using run-off transcription in a mouse cell extract as well as S1 nuclease and primer extension analysis of cellular RNA, we demonstrated that this primary processing actually results in the formation of two species of downstream RNA which differ in length by approximately 6 nucleotides, indicating the existence of two closely positioned alternative processing sites. The 200-base-pair region just 3' to the mouse processing site has a striking 80% sequence homology with a region of the human rRNA external transcribed spacer, and S1 nuclease analysis of human cellular RNA has demonstrated that an analogous rRNA processing occurs at the 5' border of the homologous human region. Unlike rDNA transcriptional initiation, however, the primary rRNA processing is not highly species specific, for the transcript of a chimeric gene containing the human processing region adjacent to a mouse rDNA promoter was synthesized and correctly processed in a mouse cell extract. This result confirms that mouse and human rRNA undergo a common primary processing event which is evidently directed by sequences within the 200-base-pair conserved sequence region.

Animals↗
PubMed1986

Treatment of Chinese hamster ovary cells with the transcriptional inhibitor actinomycin D inhibits binding of messenger RNA to ribosomes.

Inhibitors of RNA synthesis such as actinomycin D, MPB, and cordycepin progressively inhibit the initiation of protein synthesis in intact, nucleated mammalian cells. This inhibition is not dependent on the levels of mRNA, ribosomes, or tRNA. Lysates prepared from CHO cells treated with actinomycin D do not incorporate labeled globin mRNA or ovalbumin mRNA into 80S initiation complexes at the rates of untreated control extract. The ability of the extracts to produce and accumulate 48S preinitiation complexes was assessed using the 60S subunit joining inhibitors edeine and 5'-guanylyl imidodiphosphate. Control extracts were able to accumulate both the 48S preinitiation complexes and the migration-related intermediates in the presence of both inhibitors. However, lysates derived from CHO cells treated with actinomycin D were unable to produce these complexes. This was also true at low temperature, a condition that does not inhibit mRNA binding but prevents migration of the 43S complex along the mRNA. Mixing experiments with extracts from untreated control or AMD-treated CHO cells provided no evidence for a translational inhibitor. Thus, our data are consistent with the hypothesis that treatment of whole cells with actinomycin D inhibits protein synthesis initiation at the level of mRNA binding and not at migration or 60S subunit joining.

Animals↗
PubMed1983

Inhibition of protein synthesis in CHO cells by actinomycin D: lesion occurs after 40S initiation complex formation.

Inhibitors of RNA synthesis such as actinomycin D, 2-mercapto-1-(beta-4-pyridylethyl)benzimidazole, and cordycepin progressively inhibit the initiation of protein synthesis in intact nucleated mammalian cells independent of their effect on mRNA synthesis. The mechanism of this effect is unknown. The activity of cell-free lysates is not directly affected by these inhibitors, suggesting that their effect is indirect and requires an intact cell. However, lysates prepared from L-cells or CHO cells treated with the inhibitors do exhibit a decrease in initiation activity corresponding in magnitude to the effect seen in intact cells. Mixing experiments with lysates isolated from untreated or treated cells provide no evidence for a translational inhibitor. However, experiments analyzing the incorporation of [35S]methionine and [35S]Met-tRNAf into initiation complexes showed that while the level of labeled 40S initiation complex in lysates from treated cells was the same or higher than in control lysates, the rate or efficiency of formation of the 80S initiation complex was inhibited. These results imply that the transcriptional inhibitors do not affect the level or charging of the initiation tRNAMet, the activity of the eIF-2 initiation factor needed for ternary complex formation, and the availability of active 40S ribosomal subunits. Thus, this site of action is different from that observed in other translational control systems such as the hemin response in reticulocytes and the interferon-induced translation inhibition in virally infected cells. This effect may reflect the cell's coordination of nuclear transcription and cytoplasmic translation.

Adenosine Triphosphate↗
PubMed1979

Effect of temperature on protein and immunoglobulin synthesis and secretion in two mouse myeloma cell lines.

Protein synthesis in differentiated MOPC-21 and MPC-11 mouse myeloma cells was studied to determine the basis for the differences in the temperature and actinomycin D sensitivity of translation between non-differentiated mouse L-cells and differentiated rabbit reticulocytes. The temperature dependence of total protein synthesis was similar to that of L-cells and reticulocytes, being biphasic in Arrhenius plots with apparent activation energies of approximately 25 and 42 kcal/mol, above and below 25 degress C. The dependence of the secretion process was different since it was not biphasic, having a single activation energy of about 22 kcal/mol. Myeloma polysomes were like L-cell polysomes in their response to lower temperature and reached a minimum level of 50% at 15 degress C. This response was also found for the specific polysomes synthesizing the IgG H- and L-chains. In the presence of actinomycin D, myeloma polysomes declined exponentially with a half-life of approximately 6 hours. These two L-cell-like responses were not found in reticulocytes. Translation of both the IgG mRNAs and the non-IgG mRNAs was reduced by lower temperatures and actinomycin D, even though the L-chain mRNA was slightly more resistant, suggesting that this mRNA is slightly more efficient. The results of these experiments suggest that the translational differences between L-cells and reticulocytes are not mRNA dependent, but are cell type differences.

Animals↗
PubMed1975

Regulation of translation in rabbit reticulocytes and mouse L-cells; comparison of the effects of temperature.

Various parameters of protein synthesis were analyzed in rabbit reticulocytes exposed to various temperatures for up to five hours. Between 10 degrees C and 40 degrees C total protein synthesis exhibited two different apparent activation energies (36 kcal/mole, 10-24 degrees C; 22 kcal/mole, 24-40 degrees C), as did protein elongation and release (35 kcal/mole, 10-25 degrees C; 12 kcal/mole, 25-40 degrees C). However, the level of polysomes remained essentially unchanged between 0 degrees C and 42 degrees C which implies that the activation energy for polypeptide initiation is quite similar to that for elongation and is also biphasic. This situation is different from that in cultured mouse L-cells where the polysome level is dependent on temperatures. Nevertheless, reticulocytes and L-cells appear to be similar in their temperature dependence of initiation and in their rate of elongation (5-6 amino acids/second at 36 degrees C.

Animals↗
PubMed1975

Effect of reduced temperatures on protein synthesis in mouse L cells.

The rate of incorporation of leucine into protein, the rate of polypeptide elongation and termination, and the relative quantity and size of polysomes were analyzed in mouse L cells grown in suspension culture at various temperatures between 0 degrees C and 36 degrees C. Between 10 degrees C and 36 degrees C protein synthesis exhibited two different apparent activation energies (39 kcal/mole, 10-25 degrees C; 14 kcal/mole, 25-36 degrees C), whereas elongation and termination had only one (16 kcal/mole). Below 36 degrees C, the polysome level and size decreased, reaching a minimum of 30% of the control 36 degrees C values at 10 degrees C; below 10 degrees C the level increased again back to control values at 0 degrees C. The polysome decline was time dependent, requiring about 5 hr to reach the equilibrium value. This decline is completely reversible within 60 min, even in the presence of 4 mug/ml of actinomycin D, and even after 15 hr of incubation at the lower temperature. The results suggest that polypeptide initiation is rate limiting, particularly below 25 degrees C; whereas above this temperature, elongation or perhaps some other process may be limiting. These results are quite different from those obtained for E. coli and rabbit reticulocyte protein synthesis.

Animals↗