1H-NMR and protection studies of interactions between ligands and bovine pancreatic phospholipase A2.

A number of long-chain amines and naphthylamine sulfonates have been studied for their ability to inhibit bovine pancreatic phospholipase A2 (PLA2) and to protect PLA2 against alkylation of the active site histidine by p-bromophenacyl bromide. Their areas of interaction on the enzyme were further delineated using observations of chemical shift changes of assigned aromatic signals in the 1H-NMR spectrum of PLA2, while the bound conformations of two amine inhibitors were revealed using transferred nuclear Overhauser effects. The alkyl amines bind rather non-specifically on the surface of the enzyme, over the active site cleft and the interface recognition site.

Evaluation of the A&D UA-767 and Welch Allyn Spot Vital Signs noninvasive blood pressure monitors using a blood pressure simulator.

The performance of five units of the A&D UA-767 NIBP monitor and five units of the Welch Allyn Spot Vital Signs noninvasive blood pressure monitor was evaluated with the Biotek BP Pump blood pressure simulator under a variety of conditions. Using the simulator to provide a normal blood pressure waveform at 80 bpm over a range of pressures, it was found that the mean bias for the combined results from the A&D monitors was 1.9+/-2.8 mmHg and from the Welch Allyn monitors was 0.7+/-2.4 mmHg. No individual measurement showed a bias greater than 10 mmHg. A bias of greater than 5 mmHg was present in 28 out of 150 measurements for the A&D monitor and 10 out of 150 measurements for the Welch Allyn monitor. These results are comparable with ratings achieved by the instruments when tested previously according to the British Hypertension Society protocol, but testing with a simulator allowed assessment of aspects of performance which were not included in the British Hypertension Society protocol.

Potent selective inhibitors of protein kinase C.

A series of potent, selective inhibitors of protein kinase C has been derived from the structural lead provided by the microbial broth products, staurosporine and K252a. Our inhibitors block PCK in intact cells (platelets and T cells), and prevent the proliferation of mononuclear cells in response to interleukin 2 (IL2).

A novel conformationally restricted protein kinase C inhibitor, Ro 31-8425, inhibits human neutrophil superoxide generation by soluble, particulate and post-receptor stimuli.

A novel, bis-indolylmaleimide, Ro 31-8425, bearing a conformationally restricted side chain, inhibits protein kinase C isolated from rat brain and human neutrophils with a high degree of selectivity over cAMP-dependent kinase and Ca2+/calmodulin-dependent kinase. It also inhibits phorbol ester-induced intracellular events known to be mediated by protein kinase C (p47 phosphorylation in intact platelets, CD3 and CD4 down-regulation in T-cells). Ro 31-8425 inhibited superoxide generation in human neutrophils activated by both receptor stimuli (formyl-methionyl-leucylphenylalanine, opsonized zymosan, IgG and heat aggregated IgG) and post-receptor stimuli (1,2-dioctanoylglycerol and fluoride). The compound also blocked antigen driven, but not IL-2 induced, T-cell proliferation. These results support a central role for protein kinase C in the activation of the respiratory burst and antigen-driven T-cell proliferation.

4-thiophenoxy-N-(3,4,5-trialkoxyphenyl)pyrimidine-2-amines as potent and selective inhibitors of the T-cell tyrosine kinase p56lck.

We have developed a series of 4-thiophenoxy-N-(3,4,5-trialkoxyphenyl) pyrimidine-2-amines as potent and selective inhibitors of p56lck tyrosine kinase activity. In particular, the most potent inhibitor shows cellular activity in T-cell receptor (TCR) stimulated models of cytokine release, which suggests an immunomodulatory role for this class of inhibitor.

Inhibitors of protein kinase C. 3. Potent and highly selective bisindolylmaleimides by conformational restriction.

The protein kinase inhibitor staurosporine has been used to design a series of selective bisindolylmaleimide inhibitors of protein kinase C (PKC). Guided by molecular graphics, conformational restriction of the cationic side chain has led to ATP competitive inhibitors of improved potency and selectivity. Two compounds have been further evaluated and were shown to inhibit PKC of human origin and prevent T-cell activation in a human allogeneic mixed lymphocyte reaction. One of these compounds was orally absorbed in mice and antagonized a phorbol ester induced paw edema in a dose-dependent manner. This compound also selectively inhibited the secondary T-cell mediated response in a developing adjuvant arthritis model in rats and provides evidence for the potential use of PKC inhibitors as therapeutic immunomodulators.

Inhibitors of protein kinase C. 1. 2,3-Bisarylmaleimides.

The design and synthesis of a series of novel inhibitors of protein kinase C (PKC) is described. These 2,3-bisarylmaleimides were derived from the structural lead provided by the indolocarbazoles, staurosporine and K252a. Optimum activity required the imide NH, both carbonyl groups, and the olefinic bond of the maleimide ring. 2,3-Bisindolylmaleimides were the most active, and the potency of these was improved by a chloro substituent at the 5-position of one indole ring (compound 28, IC50 0.11 microM). In a series of (phenylindolyl)maleimides, nitro compound 74 was most active (IC50 0.67 microM). Naphthalene 19 and benzothiophene 21 showed greater than 100-fold selectivity for inhibition of PKC over the closely related cAMP-dependent protein kinase (PKA).

Inhibitors of protein kinase C. 2. Substituted bisindolylmaleimides with improved potency and selectivity.

A hypothetical mode of inhibition of protein kinase C (PKC) by the natural product staurosporine has been used as a basis for the design of substituted bisindolylmaleimides with improved potency over the parent compound. Structure-activity relationships were consistent with the interaction of a cationic group in the inhibitor with a carboxylate group in the enzyme, and the most potent compound had a Ki of 3 nM. The inhibitors were competitive with ATP but inhibited cAMP-dependent protein kinase (PKA) only at much higher concentrations despite the extensive sequence homology between the ATP-binding regions of PKA and PKC. Three compounds were evaluated further and found to inhibit a human allogeneic mixed lymphocyte reaction pointing to the potential utility of PKC inhibitors in immunosuppressive therapy. One of these compounds was orally absorbed in the rat and represents an attractive lead in the development of PKC inhibitors as drugs.

Novel, potent and selective inhibitors of protein kinase C show oral anti-inflammatory activity.

Clarification of the precise role of protein kinase C (PKC) in cellular functional responses has been hampered by a lack of potent, selective inhibitors. The structural lead provided by staurosporine, a potent but non-selective protein kinase (PK) inhibitor, was used to derive a series of bis(indolyl)maleimides of which the most potent, Ro 31-8425 (I50: PKC = 8 nM) showed 350-fold selectivity for PKC over cAMP-dependent protein kinase. Ro 31-8425 antagonised cellular processes triggered by phorbol esters (potent, specific PKC activators) and inhibited the allogeneic mixed lymphocyte reaction, suggesting a role for PKC in T-cell activation. Methylation of the primary amine in Ro 31-8425 produced an analogue. Ro 31-8830 which, when administered orally, produced a dose-dependent inhibition of a phorbol ester-induced paw oedema in mice (minimum effective dose = 15 mg/kg). Ro 31-8830 also selectively inhibited the secondary inflammation in a developing adjuvant arthritis model in the rat. The results presented here suggest that these selective inhibitors of PKC may have therapeutic value in the treatment of T-cell-mediated autoimmune diseases.

Relations between children's playground and classroom behaviour.

In this within-subjects design field experiment children's behaviour in the playground was charted as a function of gender and time in the classroom immediately preceding playtime. Playground behaviour was also related to post-recess classroom behaviour. Twenty-three 9-year-old children were observed for 14 weeks. Classroom behaviour (i.e., task relevant behaviour on standardised seat work immediately before and after playtime) and playground behaviour (i.e., social and non-social exercise and sedentary behaviour) were observed. Time in classroom before playtime was manipulated so that there was a shorter and a longer confinement period. Results indicated that children were less attentive to seat work as a function of time and that longer confinement resulted in more exercise for boys and more social sedentary behaviour for girls. Social behaviour at playtime and post-playtime attention to seat work were significantly related. Results are discussed in terms of Novelty Theory.

Enhancing multicultural harmony: ten actions for managers.

Health-care managers have a responsibility to create harmony within the workforce, increasingly a culturally diverse group. Suggestions offered by the literature for actions a manager can take to deal effectively with cultural diversity may not agree with employee perspectives on this issue. This article introduces a quasi-experimental evaluation method that may be used as introductory work for research in multicultural-workfoce management. This major challenge for health care in the next decade is second in importance only to health-care reform.

The use of a micro-computer in anaesthetic teaching.

The use of a relatively inexpensive microcomputer in computer-assisted learning is described. Circuits have been developed to allow slides to be shown during the course of a computer-assisted learning presentation, and also to allow the computer to control the delivery of an audiotape slide lecture.

Micro computer control of a video cassette lecture.

Circuits have been developed to allow the presentation of a video cassette lecture to be controlled by an inexpensive micro computer. The potential of both video cassette lecture and standard computer assisted learning techniques is extended, resulting in increased interaction between the student and the teaching material presented to him.

Anaesthetic pollution in the dental out-patient surgery.

Five factors influencing pollution in the dental surgery are considered. Despite leakage from round the mask and from the mouth, scavenging can remove the majority of expired anaesthetic. The effectiveness of scavenging can usefully be monitored with a Wright respirometer. Spillage of anaesthetic vapour at induction and at the end of anaesthesia must be avoided. In addition to scavenging, pollution levels are minimised if a Boyle's type machine is used and excessive fresh gas flow avoided. Room ventilation, e.g. extract fans in windows, is vital in reducing pollution levels in the surgery. Room size and non-uniformity of ventilation must also be considered.

Tapered connectors in patient breathing systems.

Investigations using recommended gauging tools have shown that many tapered connectors on breathing systems do not meet the relevant standard. It would be important to improve dimensional tolerances so that better interchangeability is achieved. This improvement may make it unnecessary to employ 'anti-disconnexion' fittings.

The microcomputer in self-assessment for examinations in anaesthesia.

A series of 30 multiple choice questions were modified for presentation on a microcomputer which was programmed to present them to the user and mark them. In a preliminary study, 15 trainee anaesthetists all agreed the system was satisfactory as an aid to preparation for the FFARCS.