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At least 19 recordsLinked to original sources
PubMed1975

Rapid analysis of the central nervous system stimulants, amphetamines, via gas chromatography-mass spectrometry: rapid acylation in the presence of a mercury catalyst.

A simple, direct, instantaneous, and quantitative acetylating procedure for amphetamines has been developed. The method uses trifluoroacetic anhydride in the presence of mercuric trifluoroacetate as a catalyst. The product was characterized by a gas chromatography-mass spectrometry system. The salient features of this technique are: (a) rapidity of analysis; (b) no previous cleanup to remove adulterants; and (c) detection of amphetamines at the picomole level. We have routinely applied this technique for court samples and law enforcement authorities. The method could prove to be applicable, with minimum amounts of involvement in sample preparation, to the analysis of a variety of compounds containing an acyl displaceable hydrogen.

Acylation↗
PubMed1979

The mechanism of action of xanthine oxidase. The relationship between the rapid and very rapid molybdenum electron-paramagnetic-resonance signals.

On the basis of the work of Gutteridge, Tanner & Bray [Biochem. J. (1978) 175, 887-897] and of other data in the literature, a mechanism for the reaction of xanthine oxidase with reducing substrates is proposed. In the Michaelis complex, xanthine is bound to molybdenum via the N-9 nitrogen atom. Coupled transfer of two electrons to molybdenum and the C-8 proton to the enzyme yields (Enzyme)-Mo-SH. Concerted with this process, reaction of the xanthine residue with a nucleophile in the active centre yields a covalent intermediate that breaks down to give the product by alternative pathways at high and at low pH values.

Chemical Phenomena↗
PubMed2026

Performance comparison of rapid and native barcoding methods for Oxford Nanopore sequencing of Poliovirus Viral Protein 1 (VP1) amplicons.

Accurate and timely sequencing of poliovirus is critical for global eradication efforts, particularly for molecular epidemiology based on the typing region of the genome, viral protein 1 (VP1). While Oxford Nanopore Technologies (ONT) sequencing has expanded capabilities for poliovirus surveillance, the relative performance of different ONT library preparation methods, including ligation-based (Native Barcoding) and transposase-based (Rapid Barcoding) approaches, has not been systematically evaluated. In this study, we compared rapid barcoding and native barcoding workflows for sequencing VP1 amplicons from 17 type 2 poliovirus-positive samples, each processed in triplicate. Native barcoding generated significantly more sequencing output, producing approximately 2.3-fold greater total read yield than rapid barcoding, and demonstrated higher run-to-run reproducibility (R2 = 0.979-0.998 vs. 0.847-0.929, respectively; p&#x202f;<&#x202f;0.001). In addition, native barcoding generated 80% of the total yield achieved by rapid barcoding within approximately 7&#x202f;h, whereas rapid barcoding required approximately 40&#x202f;h to reach the same output. Despite these differences, both methods produced identical VP1 consensus sequences across all samples, with comparable read quality (median per-base Q-scores of approximately Q17-Q18). Rapid barcoding provided substantial practical advantages, reducing hands-on library preparation time (55 vs. 200&#x202f;min) and per-sample cost ($12.82 vs. $16.54), while simplifying workflow and reducing technical complexity. These findings indicate that sequencing yield may not be a determinant of downstream analytical outcomes for poliovirus VP1 ONT sequencing. Rapid barcoding therefore represents a cost-effective and efficient approach for routine poliovirus surveillance, whereas native barcoding remains advantageous in applications requiring rapid data generation or maximal sequencing depth.

Poliovirus↗
PubMed1977

Characteristics of sarcoplasmic reticulum from slowly glycolysing and from rapidly glycolysing pig skeletal muscle post mortem.

The composition and function of fragmented sarcoplasmic reticulum from pig skeletal muscle was examined in the period immediately post mortem. Muscle was defined as being either slowly glycolysing or rapidly glycolysing on the basis of colour, pH and concentrations of glycogen and lactate. The microsomal fraction was separated on a discontinuous gradient of 35, 40 and 45% (w/v) sucrose into heavy and intermediate fractions which sedimented to the interfaces, and a light fraction which remained on the surface of the 35%-sucrose layer. The sarcoplasmic reticulum from rapidly glycolysing muscle had a lower buoyant density than had that from slowly glycolysing muscle. This was reflected in the consistent lack of material in the heavy fraction and a greater proportion in the light fraction. The latter material had significantly lower ratios (w/w) of protein to phospholipid (2.3:1 versus 3.8:1) and of protein to cholesterol (10.4:1 versus 15.6:1). There were no gross differences in phospholipid content or in fatty acid composition of individual phospholipid classes in the membranes from the two types of muscle. Analysis of membrane proteins by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis showed that ATPase (adenosine triphosphatase) was a major component of each fraction and that its contribution to the total protein content of the membrane was greater in rapidly glycolysing muscle, suggesting a loss of non-ATPase proteins. The two fractions of sarcoplasmic reticulum prepared from rapidly glycolysing muscle had approximately one-third the normal activities of Ca(2+) binding and Ca(2+) uptake in the presence of ATP and one-half the passive Ca(2+)-binding capacity in the absence of ATP of the fractions from slowly glycolysing muscle. However, the (Ca(2+)+Mg(2+))-stimulated ATPase activities were similar. Efflux from actively loaded vesicles, after the addition of EDTA, consisted of a rapid and a slow phase. Vesicles from rapidly glycolysing muscle lost 60% of associated Ca(2+) (approx. 0.10mumol of Ca(2+)/mg of protein) during the rapid phase, compared with 30% (approx. 0.17mumol of Ca(2+)/mg of protein) in those from slowly glycolysing muscle. The efflux rate during the slower phase was comparable in both types of vesicles. Analysis of the temperature-dependence of (Ca(2+)+Mg(2+))-stimulated ATPase activity revealed that a high-activation-energy process operating in the temperature range 31-45 degrees C in the intermediate and light fractions from slowly glycolysing muscle was not apparent in vesicles from rapidly glycolysing muscle. Conditions that result in the prolonged activation of glycogenolysis in pig muscle post mortem primarily affect the protein components of the sarcoplasmic-reticular membrane, giving rise to a loss of loosely associated proteins. The function of the membranes observed under these conditions does not appear to be due to enhanced permeability of the membrane to Ca(2+) and may be the result of a defect in the transport of Ca(2+) into the vesicles.

Adenosine Triphosphatases↗
PubMed1979

Longitudinal contraction of isolated guinea-pig ileum induced by rapid cooling.

1. Rapid change of bath temperature from 37 degrees C to 27 degrees C and vice versa caused longitudinal contraction of the isolated guinea-pig ileum. 2. Tetrodotoxin, tropicamide, noradrenaline, isoprenaline, morphine, and the met-enkephalin analogue FK 33-824 depressed the responses or accelerated the fade of the contraction induced by rapid cooling when added after the response had reached its maximum. 3. Hexamethonium had no influence on the responses. 4. Physostigmine potentiated all responses and reversed the fade of contraction induced by rapid cooling when added after this contraction had reached its maximum. 5. The effects of rapid cooling or warming were not altered in preparations made tachyphylactic to substance P; the response to rapid warming, but not cooling, was partially inhibited under tachyphylaxis to 5-hydroxytryptamine. 6. Antazoline, phentolamine, naloxone, and indomethacin did not block the responses. 7. Capsaicin firt potentiated and subsequently depressed the responses to both rapid cooling and warming. 8. The results indicate that rapid change of bath temperature induces longitudinal contraction by excitation of postganglionic cholinergic fibres.

Animals↗
PubMed1976

The I-D fluorescence transient. An indicator of rapid energy distribution changes in photosynthesis.

The I-D transient in the chlorophyll fluorescence induction curve (kautsky effect) is investigated in the view of recently discovered rapid changes in energy distribution between the two photosystems (Schreiber, U. and Vidaver, W., FEBS Lett., in the press). Fluorescence induction curves differ appreciably depending on whether measured at lambda less than 690 nm, originating in pigment system II, or at lambda greater than 715 nm, which is in part from pigment system I. The differences occur as well in the rapid part of the induction curve (O-I-D-P) as in the slower P-S decay. Most significant changes in energy distribution are indicated in the region of the I-D dip, being induced by appropriate preillumination. The effect is studied by (a) comparing the individual fluorescence time courses at lambda less than 690 nm and lambda greather than 715, (b) plotting F less than 690 vs. F greater than 715 and (c) recording time courses of F less than 690/F greater than 715 ratios. In (a) the I and D characteristics are delayed at F greater than 715 relative to F less than 690, which is accompanied by periods close to I and D, where the two emissions follow inverse courses. In (b) the I-D dip corresponds to a loop. And in (c) it is shown that a rapid ratio decay, reflecting increasing excitation of System I pigments, is initiated before the I-D dip. These data indicate that the I-D transient is caused by a rapid switch of energy distribution in favor of System I and resulting stimulation of Q reoxidation via the electron transport chain. It is suggested that as with the slow fluorescence transients the rapid also can be understood as a composite of two different changes, (1) direct changes resulting from a switch in energy distribution, which are inverse for F less than 690 and F greater than 715, and (2) indirect changes due to stimulated Q reduction or Q oxidation, which are parallel for both emissions. The rapid ratio decay, correlated to I-D, persists and is even stimulated in the presence of electron transport inhibitors. This and the speed of the phenomenon make it improbable that the rapid energy distribution changes are affected by an ion flux-induced mechanism. It is proposed that the electrical field across the thylakoid membrane is involved in the energy switch mechanism.

Darkness↗
PubMed1976

Influence of the rate of ethanol production and accumulation on the viability of Saccharomyces cerevisiae in "rapid fermentation".

Whereas "rapid fermentation" of diluted clover honey (25 degrees Brix) fortified with yeast nutrients using 8 X 10(8) brewers' yeast cells per ml resulted in an ethanol content of 9.5% (wt/vol; 12% vol/vol) in 3 h at 30 C, death rate of the yeast cells during this period was essentially logarithmic. Whereas 6 h was required to reach the same ethanol content at 15 C, the yeast cells retained their viability. Using a lower cell population (6 X 10(7) cells/ml), a level at which the fermentation was no longer "rapid," the yeast cells also retained their viability at 30 C. Ethanol added to the medium was much less lethal than the same or less quantities of ethanol produced by the cell in "rapid fermentation." It was considered possible that ethanol was produced so rapidly at 30 C that it could not diffuse out of the cell as rapidly as it was formed. The hypothesis was postulated that ethanol accumulating in the cell was contributing to the high death rate at 30 C. It was found that the intracellular ethanol concentration reached a level of approximately 2 X 10(11) ethanol molecules/cell in the first 30 min of fermentation at 30 C. At 15 C, with the same cell count, intracellular ethanol concentration reached a level of approximately 4 X 10(10) ethanol molecules/cell and viability remained high. Also, at 30 C with a lower cell population (6 X 10(7) cells/ml), under which conditions fermentation was no longer "rapid," intracellular ethanol concentration reached a similar level (4 X 10(10) molecules ethanol/cell) and the cells retained their viability. Alcohol dehydrogenase (ADH) lost its activity in brewers' yeast under conditions of "rapid fermentation" at 30 C but retained its activity in cells under similar conditions at 15 C. ADH activity was also retained in fermentations at 30 C with cell populations of 6 X 10(7)/ml. It would appear that an intracellular level of about 5 X 10(10) ethanol molecules/cell is normal and that this level does not damage either cell viability or ADH activity. Higher intracellular ethanol concentrations, such as 2 X 10(11) molecules ethanol/cell (a fourfold increase in intracellular ethanol concentration), are accompanied by inactivation of ADH and loss of cell viability.

Alcohol Oxidoreductases↗
PubMed1977

Evaluation of the rapid hippurate hydrolysis test with enterococcal group D streptococci.

The rapid hippurate hydrolysis test was evaluated with the conventional test, using 17 group A streptococci, 9 non-enterococcal group D streptococci, 108 enterococcal group D streptococci, and 2 strains of Listeria monocytogenes. There was complete correlation between the rapid and conventional tests with all organisms except enterococcal group D. The rapid hippurate hydrolysis method was more sensitive with the enterococci; 95.4% were positive with the rapid method, and 9.3% were positive with the conventional method. Thin-layer chromatography (TLC) was performed on all isolates to determine if the end product of hydrolysis, glycine, was indeed present. The TLC results were in agreement with the rapid and conventional methods for group A streptococci, nonenterococcal group D streptococci, and L. monocytogenes. TLC results were in total agreement with the rapid hippurate hydrolysis test for the enterococcal group D isolates, thus verifying the accuracy of this more sensitive test. Trace amounts of glycine were found in the substrate, indicating the need for including an uninoculated substrate control as well as stock strains of group A and B beta-hemolytic streptococci (negative and positive controls, respectively) each time the rapid hippurate hydrolysis test is performed.

Amidohydrolases↗
PubMed1976

Patient-initiated rapid atrial pacing to manage supraventricular tachycardia.

Patient-controlled rapid atrial pacing was used to manage 12 cases of recurrent supraventricular tachycardia refractory to drug therapy. The pacing system consists of an implanted receiver-lead system and an external patient-activated transmitter. In each case, brief periods (5 to 20 seconds) of rapid atrial pacing were effective in terminating the supraventricular tachycardia and resulted in a return to normal sinus rhythm. In three patients, occasional transient episodes of atrial flutter or atrial fibrillation preceded a spontaneous return to normal sinus rhythm. The pacing system was removed in one patient 13 months postoperatively because of persistent pericarditis; one patient died of an unrelated cerebral hemorrhage 13 months postoperatively. Successful management of supraventricular tachycardia has been maintained in the 10 remaining patients for 15 to 36 months (average 26.4). In more than 6,000 patient applications of rapid atrial pacing, there has been only one failure to convert the tachycardia. Successful application of permanent rapid atrial pacing requires (1) prescreening of patients with temporary external rapid atrial pacing to verify susceptibility to conversion of supraventricular tachycardia and absence of anomalous conduction pathways that may permit conduction of rapid pacing rates to the ventricles, and (2) assessment of the patient's ability to use the transmitter properly.

Adult↗
PubMed1975

Bovine brain Na+,K+-stimulated ATP phosphohydrolase studied by a rapid-mixing technique. K+-stimulated liberation of [32P] orthophosphate from [32P] phosphoenzyme and resolution of the dephosphorylation into two phases.

Dephosphorylation of [32P]phosphoenzyme of bovine brain Na+,K+-stimulated ATP phosphohydrolase (EC 3.6.1.3), labelled by [gamma-32P]ATP, was investigated at 21 degrees C by means of a rapid-mixing technique. On addition of a high concentration of KCl (10 mM) to [32P]phosphoenzyme at steady state in the presence of Mg2+ and Na+, very rapid dephosphorylation was obtained. Simultaneously, the amount of [32P]orthophosphate increased at about the same rate. It was concluded that this K+-stimulated dephosphorylation and liberation of [32P]orthophosphate from the [32P]phosphoenzyme was rapid enough to participate in the Na+,K+-stimulated hydrolysis of ATP. In order to study the dephosphorylation in absence of continuing 32P-labelling, excess unlabelled ATP or a chelator of Mg2+ was added. Simultaneous addition of a high concentration of KCl to the [32P]phosphoenzyme formed in the presence of Mg2+ and Na+ but in the absence of K+, resulted in an initial very rapid phase and a subsequent slower phase of dephosphorylation. With KCl also initially present in the incubation medium, only the slow phase was observed. The slow phase of dephosphorylation also seemed to be sufficiently rapid to participate in the Na+, K+-stimulated ATPase reaction. On addition of a high concentration of ADP (5 mM) to [32P]phosphoenzyme formed in the presence of Mg2+ and Na+, an initial comparatively rapid, and later slow phase of dephosphorylation were detected. This gave further support for different forms of phosphoenzyme. Approximate concentrations of these forms, in the absence and presence of KCl, were estimated by extrapolation and the turnover of these forms was calculated. The nature of the kinetically different components of phosphoenzyme and their role in the Na+, K+-stimulated ATPase reaction is discussed.

Adenosine Triphosphatases↗
PubMed1977

Differential association of F' plasmid and R plasmid deoxyribonucleic acid with a rapidly sedimenting fraction of a Proteus mirabilis lysate.

We have examined the association of an F' plasmid and an R plasmid in Proteus mirabilis with a rapidly sedimenting material that is generated by sodium dodecyl sulfate lysis and low speed centrifugation. Virtually all of the chromosomal deoxyribonucleic acid (DNA) and the F' plasmid DNA are associated with the rapidly sedimenting material after gentle lysis and centrifugation. A portion of R plasmid NR1 DNA (usually 5 to 25%) is not bound to the rapidly sedimenting material and is recovered in the supernatant fraction. This difference in binding is not related to the size of the plasmid DNA, since F' plasmids and R plasmids of different molecular weights showed the same behavior. R plasmid DNA labeled by a brief pulse of [(3)H]thymine is recovered in the supernatant fraction to a lower extent than the total R plasmid DNA. It would appear that R plasmid replication takes place in association with the rapidly sedimenting material. With prolongation of the [(3)H]thymine pulse, the [(3)H]thymine-labeled R plasmid DNA is recovered in the supernatant fraction with the same probability as the total R plasmid DNA. This finding indicates that a change in R plasmid attachment to the rapidly sedimenting material occurs some time after its replication. The differences observed in the replication of F' plasmids and R plasmids in P. mirabilis may be related to their different modes of association with the rapidly sedimenting material.

Bacteriolysis↗
PubMed1979

Survival of frozen mouse embryos after rapid thawing from -196 degrees C.

The effect of the rate of rewarming on the survival of 8-cell mouse embryos and blastocysts was examined. The samples were slowly cooled (0.3--0.6 degrees C/min) in 1.5 M-DMSO to temperatures between -10 and -80 degrees C before direct transfer to liquid nitrogen (-196 degrees C). Embryos survived rapid thawing (275--500 degrees C/min) only when slow cooling was terminated at relatively high subzero temperatures (-10 to -50 degrees C). The highest levels of survival in vitro of rapidly thawed 8-cell embryos were obtained after transfer to -196 degrees C from -35 and -40 degrees C (72 to 88%) and of rapidly thawed blastocysts after transfer from -25 to -50 degrees C (69 to 74%). By contrast, for embryos to survive slow thawing (8 to 20 degrees C/min) slow cooling to lower subzero temperatures (-60 degrees C and below) was required before transfer to -196 degrees C. The results indicate that embryos transferred to -196 degrees C from high subzero temperatures contain sufficient intracellular ice to damage them during slow warming but to permit survival after rapid warming. Survival of embryos after rapid dilution of DMSO at room temperature was similar to that after slow (stepwise) dilution at 0 degrees C. There was no difference between the viability of rapidly and slowly thawed embryos after transfer to pseudopregnant foster mothers. It is concluded that the behaviour of mammalian embryos subjected to the stresses of freezing and thawing is similar to that of other mammalian cells. A simpler and quicker method for the preservation of mouse embryos is described.

Animals↗