Activity of the plasma membrane H(+)-ATPase and optimal glycolytic flux are required for rapid adaptation and growth of Saccharomyces cerevisiae in the presence of the weak-acid preservative sorbic acid

The weak acid sorbic acid transiently inhibited the growth of Saccharomyces cerevisiae in media at low pH. During a lag period, the length of which depended on the severity of this weak-acid stress, yeast cells appeared to adapt to this stress, eventually recovering and growing normally. This adaptation to weak-acid stress was not due to metabolism and removal of the sorbic acid. A pma1-205 mutant, with about half the normal membrane H+-ATPase activity, was shown to be more sensitive to sorbic acid than its parent. Sorbic acid appeared to stimulate plasma membrane H+-ATPase activity in both PMA1 and pma1-205. Consistent with this, cellular ATP levels showed drastic reductions, the extent of which depended on the severity of weak-acid stress. The weak acid did not appear to affect the synthesis of ATP because CO2 production and O2 consumption were not affected significantly in PMA1 and pma1-205 cells. However, a glycolytic mutant, with about one-third the normal pyruvate kinase and phosphofructokinase activity and hence a reduced capacity to generate ATP, was more sensitive to sorbic acid than its isogenic parent. These data are consistent with the idea that adaptation by yeast cells to sorbic acid is dependent on (i) the restoration of internal pH via the export of protons by the membrane H+-ATPase in an energy-demanding process and (ii) the generation of sufficient ATP to drive this process and still allow growth.     

Metrics and mappings: A framework for understanding real-world quantitative estimation.

Estimation is influenced by a variety of processes: application of heuristics, domain-specific reasoning, and intuitive statistical induction among them. In this article, the authors propose the metrics and mapping framework to account for how these processes are integrated to generate estimates. This framework identifies 2 types of information as critical: knowledge of distributional properties (metric knowledge) and knowledge of relative status of individual entities within the distribution (mapping knowledge). Heuristics and domain-specific knowledge are both viewed as cues that contribute to mapping knowledge; intuitive statistical induction is viewed as providing cues to metric properties. Results of 4 experiments illustrate the framework's usefulness for integrating these types of information and for predicting when people emphasize heuristics and when they emphasize domain-specific knowledge. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Inhibition of TEM-2 beta-lactamase from Escherichia coli by clavulanic acid: observation of intermediates by electrospray ionization mass spectrometry

Clavulanic acid, the therapeutically important inhibitor of beta-lactamases containing a nucleophilic serine residue at their active sites, inhibits Escherichia coli TEM-2 beta-lactamase via a complex mechanism. Electrospray ionization mass spectrometry (ESIMS) studies revealed that a minimum of four different modified proteins are formed upon incubation of clavulanate with the TEM-2 enzyme. These exhibit mass increments relative to the unmodified TEM-2 beta-lactamase of 52, 70, 88, and 155 Da. Time course studies implied that no long-lived forms of clavulanate-inhibited TEM-2 beta-lactamase retain the carbons of the oxazolidine ring of clavulanate. The absence of a 199 Da increment to unmodified TEM-2 suggests rapid decarboxylation of clavulanate upon binding to the enzyme. Proteolytic digestions of purified forms of clavulanate inhibited TEM-2 beta-lactamase followed by analyses using high-performance liquid chromatography coupled to ESIMS (HPLC-ESIMS) and chemical sequencing were used to provide positional information on the modifications to the enzyme. Increments of 70 and 80 Da increments were shown to be located in a peptide containing Ser-70. A further 70 Da mass increment, assigned as a beta-linked acrylate, was localized to a peptide containing Ser-130. A mechanistic scheme for the reaction of clavulanate with TEM-2 beta-lactamase is proposed in which acylation at Ser-70 and subsequent decarboxylation is followed either by cross-linking with Ser-130 to form a vinyl ether or by reformation of unmodified enzyme via a Ser-70 linked (hydrated) aldehyde. Purified cross-linked vinyl ether was observed to slowly convert under acidic conditions to a Ser-70 linked (hydrated) aldehyde with concomitant conversion of Ser-130 to a dehydroalanyl residue.     

Echinococcal disease and the provision of humanitarian surgical aid in Bosnia

The Canadian Armed Forced have been deployed to the republics of the former Yugoslavia since 1992 as part of the United Nations Protection Force and the NATO-led Implementation Force. Most of the combat arms units have been supported by small, self-contained surgical teams for essential surgery. Considerable benefit is gained by cooperating with civilian surgeons. The experience of treating five patients with complicated hydatid disease endemic to the area is examined. Treatment of these patients requires performing major surgical procedures under austere conditions and must be undertaken with care. Careful selection and communication with the surgical nursing team and civilian surgeons is essential. Well selected cases can also pay tremendous dividends in terms of maintaining the skills of personnel who must be prepared for any emergency in addition to providing vital surgical assistance to these patients.     

The use of capsaicin in herpes zoster ophthalmicus neuralgia

The final stage in the production of a radiotherapy treatment plan must always be an independent check that the linear accelerator settings given on the plan do in fact deliver the required dose distribution. A tool is described that enables rapid checking of diaphragm settings in relation to the patient.     

Flash photolysis of the carbon monoxide compounds of wild-type and mutant variants of cytochrome bo from Escherichia coli

The carbon monoxide compounds of the fully reduced and mixed valence forms of cytochrome bo from Escherichia coli were laser photolysed under anaerobic conditions at room temperature. The carbon monoxide recombined with characteristic rate constants of 50 s-1 or 35 s-1 in the fully reduced and mixed valence forms, respectively. Rates of CO recombination with the fully reduced enzyme were examined in a variety of mutant forms of cytochrome bo, produced by site-directed mutagenesis. A method was developed to deconvolute cytochromes bo and bd, leading to some reassessment of histidine ligands to the metals. Significant changes in the rate constant of recombination of carbon monoxide occurred in many of these mutants and these results could be rationalised generally in terms of our current working model of the folding structure of subunit I. In the mixed valence form of the enzyme the transient photolysis spectra in the visible region are consistent with a rapid electron redistribution from the binuclear centre to the low-spin haem. This electron transfer is biphasic, with rate constants of around 10(5) and 8000 s-1. The process was also examined in the His-333-Leu mutant, in which a putative histidine ligand to CuB is replaced by leucine, and which results in the loss of the CuB. It appeared that rapid haem-haem electron transfer could still occur. The observation that CuB is apparently not required for rapid haem-haem electron transfer is consistent with the recently proposed model in which the two haems are positioned on opposite sides of transmembrane helix X in subunit I of the oxidase.     

The UK Renewable Energy programme

The UK Government has supported a programme on Renewable Energy since 1975. In 1988 progress was reviewed and this led to some redirection of the programme with greater emphasis on the commercial development of renewables, as published in Energy Paper 55.The climate in which Renewable Energy operates has been influenced recently by a number of factors including privatisation of the UK electricity supply industry, when the Government introduced the New Fossil Fuel Obligation and publication of the Government's 1990 White Paper on the Environment which noted that the Government is working towards a figure of new renewable energy generating capacity of 1000 MW by 2000.The paper aims to give an overview of the present state of development of the renewable technologies in the UK and to discuss some of the factors - technical and non-technical - which will affect their likely future development.