1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine as a substrate of cytochrome P450 2D6: allosteric effects of NADPH-cytochrome P450 reductase

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), a neurotoxin that produces Parkinsonism symptoms in man, has been examined as a substrate of recombinant human cytochrome P450 2D6. When cumene hydroperoxide is used as an oxygen and electron donor, a single product is formed, identified as 4-phenyl-1,2,3,6-tetrahydropyridine. The K(m) for formation of this product (130 microM) is in agreement with the dissociation constants for MPTP binding to the enzyme determined by optical and nuclear magnetic resonance (NMR) spectroscopy. When the reaction is carried out with nicotinamide adenine dinucleotide phosphate (reduced) (NADPH) and recombinant human NADPH-cytochrome P450 reductase, a second product, identified as 1-methyl-4-(4'-hydroxyphenyl)-1,2,3,6-tetrahydropyridine, is formed in addition to 4-phenyl-1,2,3,6-tetrahydropyridine. The K(m) values for formation of these two products are 19 microM and 120 microM, respectively. Paramagnetic relaxation experiments have been used to measure distances between the protons of bound MPTP and the heme iron, and these have been used to construct models for the position and orientation of MPTP in the active site. For the cytochrome alone, a single mode of binding was observed, with the N-methyl close to the heme iron in a position appropriate for the observed N-demethylation reaction. In the presence of the reductase, the data were not consistent with a single mode of binding but could be explained by the existence of two alternative orientations of MPTP in the active site. One of these, characterized by a dissociation constant of 150 microM, is essentially identical to that observed in the absence of the reductase. In the second, which has a K(d) of 25 microM, the MPTP is oriented so that the aromatic ring is close to the heme iron, in a position appropriate for p-hydroxylation leading to the formation of the product seen only in the presence of the reductase. In the case of codeine, another substrate for cytochrome P450 2D6, the addition of reductase had no effect on the nature of the product formed, the dissociation constant, or the orientation in the binding site. These observations show that NADPH-cytochrome P450 reductase has an allosteric effect on the active site of cytochrome P450 2D6 that affects the binding of some substrates but not others.     

Tissue specific expression of FMR-1 provides evidence for a functional role in fragile X syndrome

Prostaglandin E2 levels in isolated rat islets were increased from 64 +/- 11 pg/30 islets when incubated in medium containing 2 mM glucose to 115 +/- 9 pg/30 islets in medium containing 20 mM glucose. In contrast, glyceraldehyde (10 mM) reduced prostaglandin E2 levels to 29 +/- 6 pg/30 islets. Inhibition of glucose metabolism by mannoheptulose (10 mM) abolished the stimulatory effect of glucose on prostaglandin E2 levels and inhibited glucose-induced insulin release. The cyclooxygenase inhibitor, flurbiprofen (20 microM), did not affect insulin release caused by glucose or glyceraldehyde. In the presence of 1 mg/ml bovine serum albumin, insulin secretion induced by 20 mM glucose (6.9 +/- 1.1% of islet insulin content) was reduced by the lipoxygenase inhibitor BW755 C (20 microM) to 3.1 +/- 0.6%, and by the phospholipase A2 inhibitor, p-bromophenacyl bromide (10 microM), to 2.1 +/- 0.8%. In the absence of bovine serum albumin the inhibitory action of BW755 C and p-bromophenacyl bromide on glucose-induced insulin release was significantly more pronounced. These drugs whether in the presence or absence of bovine serum albumin, did not affect glyceraldehyde-stimulated insulin secretion. Glyceraldehyde (10 mM), potentiated glucose-induced insulin release in the presence of 2-8 mM glucose, but not for 10-20 mM glucose. Although the phospholipase A2 activator, melittin, initiated insulin release in the presence of 2 mM glucose and enhanced 10 mM glyceraldehyde-stimulated insulin secretion it had no effect on 20 mM glucose-induced insulin release. These two stimulatory effects of melittin on insulin release were totally abolished by p-bromophenacyl bromide.(ABSTRACT TRUNCATED AT 250 WORDS)     

Carla: A rule language for specifying communications architectures

Due to the unique requirements of a series of projects to specify communications architectures using graphical representations (Cara and MFD), we have developed the communications-oriented rule-based language Carla (Cara Rule Language), which provides an executable specification of the architecture being developed. Carla is designed to provide the ability to specify and simulate high-level, possibly incomplete, specifications of communications architectures, and to allow the developer to refine the specification through the addition of behavior-describing rules. Carla is also well-suited to creating black-box specifications of any system whose behavior depends on input/output history. We describe the features of the language, discuss various design issues, and provide examples of various communications protocols specified in Carla.     

Climate change gains more from nuclear substitution than from conservation

We show the massive reduction achievable, in both emissions and climate change impact, from enhanced nuclear energy use on the forecasts of future world energy use and its associated environmental impacts. A range encompassing the major scenarios for the World's energy demand have been analyzed using the latest version of the climate-modeling MAGICC/SCENGEN software (Version 4.1). We have updated and predicted the impacts of 80% substitution with CO₂-free sources (likely predominantly nuclear) for coal-fired electricity (by 2030) and for transportation fuel (by 2040). For transportation, hydrogen produced by CO₂-free sources would replace gasoline and diesel fuels. In this paper, to bracket the range of futures, we simply focus on two scenarios from the Intergovernmental Panel on Climate Change's (IPCC), one (A1FI) that is energy-profligate and one (B2) that is energy-conserving.The results show that, interestingly, projected average global temperatures for all scenarios are fairly similar until about 2035 (a further rise beyond the 1990 average temperature of +0.75 ± 0.1 K) regardless of energy usage and its sources. However, by 2050, the different IPCC scenarios diverge markedly. Understandably, A1FI is projected to have noticeably stronger effects than B2 on average global temperatures (about 0.3 K more in 2050) but the effect is much stronger over land at mid and high latitudes (up to almost 1 K more). What is most striking is that the substitution of CO₂-free sources gives projected average temperature rises in 2050 over key land areas (North America and China) that are very similar for the two energy-use scenarios—typically 1–1.5 K because A1FI's additional energy is predominantly supplied by nuclear. In contrast, projected rises with the unaltered cases are markedly different being about 2.5 K for A1FI and 1.5–2 K for B2. The projected changes in rainfall distribution show similar patterns, especially for the expected increases in higher latitudes.With the assumption of no additional policies for substitution of energy sources beyond 2040, temperature divergence between the two scenarios of relative energy profligacy or conservation grows in the latter half of the 21st century, even with substitution. However, the early substitution of nuclear energy and hydrogen appears to buy time and is not crucially dependent on severe, near-term curtailment of energy use. Near-term curtailment is too difficult to implement at a time of rapid industrialization of major emerging economies. Of course, proportionately larger deployments of CO₂-free energy sources are needed for more energy-intensive scenarios.Nuclear power must dominate as the source of CO₂-free energy since it is proven, dependable, available on a large scale, and economic. Social objections to nuclear energy in some countries and quarters are seen as well-meaning but misguided distractions from solving the energy and environmental crises that are now facing world sustainability. The time for real technical, social and political action is now.     

Mixing and solid-liquid mass-transfer rates in a creusot-loire uddeholm vessel: A water model case study

The process of mixing and solid-liquid mass transfer in a one-fifth scale water model of a 100-ton Creusot-Loire Uddeholm (CLU) converter was investigated. The modified Froude number was used to relate gas flow rates between the model and its protoype. The influences of gas flow rate between 0.010 and 0.018 m³/s and bath height from 0.50 to 0.70 m on mixing time were examined. The results indicated that mixing time decreased with increasing gas flow rate and increased with increasing bath height. The mixing time results were evaluated in terms of specific energy input and the following correlation was proposed for estimating mixing times in the model CLU converter: T _mix=1.08Q ^−1.05 W ^0.35, where Q (m³/s) is the gas flow rate and W (tons) is the model bath weight. Solid-liquid mass-transfer rates from benzoic acid specimens immersed in the gas-agitated liquid phase were assessed by a weight loss measurement technique. The calculated mass-transfer coefficients were highest at the bath surface reaching a value of 6.40 × 10⁻⁵ m/s in the sprout region. Mass-transfer coefficients and turbulence parameters decreased with depth, reaching minimum values at the bottom of the vessel.     

A specific,sensitive and high-capacity immunoassay for PAF

A specific radioimmunoassay for platelet-activating factor (PAF) sensitive in the range 10–1000 pg (0.02–2 pmoles) has been developed. Detailed quantitative hapten inhibition studies showed specificity for the acetyl group at C-2 of PAF, a requirement for the ether linkage at C-1 and some tolerance for substituents on the choline nitrogen. No significant cross-reactivity was found with phosphatidylcholine and lysophosphatidylcholine or with lysoPAF. Based on a paper presented at the Third International Conference on Platelet-Activating Factor and Structurally Related Alkyl Ether Lipids, Tokyo, Japan, May 1989.     

Traumatic aniridia after small incision cataract extraction.

INTRODUCTION: Phacoemulsification and falls are both common in the elderly population. We present a case of acquired total aniridia and vitreous haemorrhage occurring as a result of trauma in a pseudophakic eye. METHODS: Interventional case report with history, clinical photograph and discussion with literature review. RESULTS: A previously healthy 74-year-old Caucasian female was referred with a painful left eye and poor vision following a fall and trauma to the left side of her face. Initial examination confirmed visual acuity of perception of light with total hyphaema and vitreous haemorrhage. Subsequent examination revealed complete aniridia with an intact capsular bag and well-centred posterior chamber intraocular lens and attached retina. Final visual acuity after treatment was 6/9. CONCLUSIONS: Blunt trauma may cause total iris disinsertion in and expulsion from pseudophakic eyes. This relatively novel injury may present increasingly commonly to casualty departments and ophthalmologists.     

Optimal control of nonlinear systems to given orbits

Using optimal control techniques we derive and demonstrate the use of an explicit single-step control method for directing a nonlinear system to a target orbit and keeping it there. We require that control values remain near the uncontrolled settings. The full nonlinearity of the problem in state space variables is retained. The “one-step” of the control is typically a composition of known or learned maps over (a) the time required to learn the state, (b) the time to compute the control and (c) the time to apply the control. No special targeting is required, yet the time to control is quite rapid. Working with the dynamics of a well-studied nonlinear electrical circuit, we show how this method works efficiently and accurately in two situations: when the known circuit equations are used, and when control is performed only on a Poincaré section of the reconstructed phase space. In each case, because the control rule is known analytically, the control strategy is computationally efficient while retaining high accuracy. The target locations on the selected target trajectory at each control stage are determined dynamically by the initial conditions and the system dynamics, and the target trajectory is an approximation to an unstable periodic orbit of the uncontrolled system. A linear stability analysis shows that dissipation in the dynamical system is essential for reaching a controllable state.     

The CADE-14 ATP System Competition

This paper documents the design, competing systems, results, and conclusions of the CADE-14 ATP System Competition (CASC-14).     

Using formalized temporal message-flow diagrams

Temporal message-flow diagrams (TMFDs), alternatively called sequence charts, interaction diagrams, event traces, or actor diagrams, are illustrations of a system's global message-passing activity over time, and a pictorial aid to understanding the system's behavior. They are widely used for requirements and documentation for network protocols and object-oriented applications. We present a general formalism for TMFDs, describe a suite of tools we have designed that employs this formalism, and present our experiences with these tools. The formalism and tools described serve to support and broaden the use of TMFDs in developing communicating systems.