Characterization of the sequential non-covalent and covalent interactions of the antitumour antibiotic hedamycin with double stranded DNA by NMR spectroscopy

Hedamycin, a member of the pluramycin class of antitumour antibiotics, consists of a planar anthrapyrantrione chromophore to which is attached two aminosugar rings at one end and a bisepoxide-containing sidechain at the other end, Binding to double-stranded DNA is known to involve both reversible and non-reversible modes of interaction. As a part of studies directed towards elucidating the structural basis for the observed 5'-pyGT-3' sequence selectivity of hedamycin, we conducted one-dimensional NMR titration experiments at low temperature using the hexadeoxyribonucleotide duplexes d(CACGTG)(2) and d(CGTACG)(2). Spectral changes which occurred during these titrations are consistent with hedamycin initially forming a reversible complex in slow exchange on the NMR timescale and binding through intercalation of the chromophore. Monitoring of this reversible complex over a period of hours revealed a second type of spectral change which corresponds with formation of a non-reversible complex. Copyright (C) 1999 John Wiley & Sons, Ltd.

Sequential increase in Egr-1 and AP-1 DNA binding activity in the dentate gyrus following the induction of long-term potentiation

Establishment of long-term potentiation (LTP) at perforant path synapses is highly correlated with increased expression of Egr and AP-1 transcription factors in rat dentate gyrus granule cells. We have investigated whether increased transcription factor levels are reflected in increased transcription factor activity by assessing Egr and AP-I DNA binding activity using gel shift assays. LTP produced an increase in binding to the Egr element, which was NMDA receptor-dependent and correlated closely with our previously reported increase in Egr-1 (zif/268) protein levels. Supershift analysis confirmed involvement of Egr-1, but not Egr-2 in the DNA binding activity. AP-1 DNA binding was also rapidly elevated in parallel with protein levels, however, the peak increase in activity was delayed until 4 h, a time point when we have previously shown that only jun-D protein was elevated. These data indicate that binding of Egr-1 and AP-1 to their response elements is increased in two phases. This may result in activation of distinct banks of target genes which contribute to the establishment of persistent LTP. (C) 2000 Elsevier Science B.V. All rights reserved.

Influence of patient age and implantation technique on the probability of re-replacement of the homograft aortic valve

Background and aim of the study: Results of valve re-replacement (reoperation) in 898 patients undergoing aortic valve replacement with cryopreserved homograft valves between 1975 and 1998 are reported. The study aim was to provide estimates of unconditional probability of valve reoperation and cumulative incidence function (actual risk) of reoperation. Methods: Valves were implanted by subcoronary insertion (n = 500), inclusion cylinder (n = 46), and aortic root replacement (n = 352). Probability of reoperation was estimated by adopting a mixture model framework within which estimates were adjusted for two risk factors: patient age at initial replacement, and implantation technique. Results: For a patient aged 50 years, the probability of reoperation in his/her lifetime was estimated as 44% and 56% for non-root and root replacement techniques, respectively. For a patient aged 70 years, estimated probability of reoperation was 16% and 25%, respectively. Given that a reoperation is required, patients with non-root replacement have a higher hazard rate than those with root replacement (hazards ratio = 1.4), indicating that non-root replacement patients tend to undergo reoperation earlier before death than root replacement patients. Conclusion: Younger patient age and root versus non-root replacement are risk factors for reoperation. Valve durability is much less in younger patients, while root replacement patients appear more likely to live longer and hence are more likely to require reoperation.

The initial effects of a Mulligan's mobilization with movement technique on dorsiflexion and pain in subacute ankle sprains

Physiotherapists frequently use manipulative therapy techniques to treat dysfunction and pain resulting from ankle sprain. This study investigated whether a Mulligan's mobilization with movement (MWM) technique improves talocrural dorsiflexion, a major impairment following ankle sprain, and relieves pain in subacute populations. Fourteen subjects with subacute grade II lateral ankle sprains served as their own control in a repeated measures, double-blind randomized controlled trial that measured the initial effects of the MWM treatment on weight bearing dorsiflexion and pressure and thermal pain threshold. The subacute ankle sprain group studied displayed deficits in dorsiflexion and local pressure pain threshold in the symptomatic ankle. Significant improvements in dorsiflexion occurred initially post-MWM (F-(2,F-26) 7.82, P = 0.002), but no significant changes in pressure or thermal pain threshold were observed after the treatment condition. Results indicate that the MWM treatment for ankle dorsiflexion has a mechanical rather than hypoalgesic effect in subacute ankle sprains. The mechanism by which this occurs requires investigation if we are to better understand the role of manipulative therapy in ankle sprain management. (C) 2003 Elsevier Ltd. All rights reserved.

Prediction of impact forces in a vibratory ball mill using an inverse technique

To understand the dynamic mechanisms of the mechanical milling process in a vibratory mill, it is necessary to determine the characteristics of the impact forces associated with the collision events. However, it is difficult to directly measure the impact force in an operating mill. This paper describes an inverse technique for the prediction of impact forces from acceleration measurements on a vibratory ball mill. The characteristics of the vibratory mill have been investigated by the modal testing technique, and its system modes have been identified. In the modelling of the system vibration response to the impact forces, two modal equations have been used to describe the modal responses. The superposition of the modal responses gives rise to the total response of the system. A method based on an optimisation approach has been developed to predict the impact forces by minimising the difference between the measured acceleration of the vibratory ball mill and the predicted acceleration from the solution of the modal equations. The predicted and measured impact forces are in good agreement. Copyright (C) 1996 Elsevier Science Ltd.

Supersaturated solutions evaluated with an in vitro stratum corneum tape stripping technique

The flux of a compound across a membrane from any formulation, whether it contains penetration enhancers or not, is limited by its saturated solubility in the vehicle. Under such conditions the concentration of the permeant in the outer layers of the stratum corneum is also saturated. Consequently, when the permeation of a drug from a supersaturated solution leads to enhanced penetration, the concentration of the drug in the outer layers of the membrane is also supersaturated. Therefore, the stratum corneum may possess antinucleant properties which inhibit or retard the crystallisation process. In this study, the enhanced in vitro permeation of supersaturated solutions of piroxicam across human skin in diffusion cells was demonstrated. The amount of permeant in the stratum corneum was determined using a tape stripping technique. Supersaturated solutions up to four degrees of saturation were investigated which produced a linear relationship between the degree of saturation and the amount of piroxicam in the stratum corneum (R-2 = 0.970). Furthermore, the amount of piroxicam in the viable layers of the skin also increased with increasing degree of saturation. An analysis of the results suggested that enhanced penetration across human skin from supersaturated solutions of piroxicam may occur as a result of the antinucleating ability of the intercellular lipids of the stratum corneum. (C) 1997 Elsevier Science B.V.

Sequential asymmetric auctions with endogenous participation

In this paper we suggest a model of sequential auctions with endogenous participation where each bidder conjectures about the number of participants at each round. Then, after learning his value, each bidder decides whether or not to participate in the auction. In the calculation of his expected value, each bidder uses his conjectures about the number of participants for each possible subgroup. In equilibrium, the conjectured probability is compatible with the probability of staying in the auction. In our model, players face participation costs, bidders may buy as many objects as they wish and they are allowed to drop out at any round. Bidders can drop out at any time, but they cannot come back to the auction. In particular we can determine the number of participants and expected prices in equilibrium. We show that for any bidding strategy, there exists such a probability of staying in the auction. For the case of stochastically independent objects, we show that in equilibrium every bidder who decides to continue submits a bid that is equal to his value at each round. When objects are stochastically identical, we are able to show that expected prices are decreasing.

The validity of self-reported energy intake as determined using the doubly labelled water technique

In the 1980s the development of the doubly labelled water (DLW) technique made it possible to determine the validity of dietary assessment methods using external, independent markers of intake in free-living populations. Since then, the accuracy of self-reported energy intake (EI) has been questioned on a number of occasions as under-reporting has been found to be prevalent in many different populations. This paper is a review of investigations using the DLW technique in conjunction with self-reported EI measures in groups including adults, children and adolescents, obese persons, athletes, military personnel and trekking explorers. In studies where a person other than the subject is responsible for recording dietary intake, such as parents of young children, EI generally corresponds to DLW determined energy expenditure. However, in instances where the subjects themselves report their intake, EI is generally under-reported when compared with energy expenditure. It was originally believed that this phenomenon of under-reporting was linked to increased adiposity and body size, however, it is now apparent that other factors, such as dietary restraint and socio-economic status, are also involved. This paper therefore aims to present a more comprehensive picture of under-reporting by tying in the findings of many DLW studies with other studies focusing particularly on the characteristics and mechanisms for under-reporting. Awareness of these characteristics and mechanisms will enable researchers to obtain more accurate self-reports of EI using all dietary recording techniques.

A technique for preserving pigmentation in some capsalid monogeneans for taxonomic purposes

A technique is described to preserve the pigment found in the bodies and the intestine of some brightly coloured and darkly pigmented benedeniine capsalid monogeneans. Previous studies of these pigmented capsalids have proven difficult because the pigmentation usually disappears when the worms are fixed using preservatives containing concentrations of formalin over 5% and/or ethanol, acetic acid, chromic acid, picric acid and mercuric chloride. The technique developed here uses a fixative comprising glycerol, acetone and formalin (GAF). After fixation under light coverslip compression for three minutes, specimens are transferred to absolute acetone for three minutes and cleared in a mixture of nine parts cedar wood oil and one part absolute acetone before mounting in Canada balsam. Processing must be carried out quickly, as these chemicals will cause the pigments to fade if the specimens are exposed to them for too long. Pigmented benedeniines processed using this technique retain the distribution, intensity and colour observed in live worms. The colour and distribution of pigmentation in monogeneans may be of taxonomic importance and this technique aids preparation of whole-mounts suitable for registration as type-material.

Analysis of the gate capacitance measurement technique and its application for the evaluation of hot-carrier degradation in submicrometer MOSFETs

The use of gate-to-drain capacitance (C-gd) measurement as a tool to characterize hot-carrier-induced charge centers in submicron n- and p-MOSFET's has been reviewed and demonstrated. By analyzing the change in C-gd measured at room and cryogenic temperature before and after high gate-to-drain transverse field (high field) and maximum substrate current (I-bmax) stress, it is concluded that the degradation was found to be mostly due to trapping of majority carriers and generation of interface states. These interface states were found to be acceptor states at top half of band gap for n-MOSFETs and donor states at bottom half of band gap for p-MOSFETs. In general, hot electrons are more likely to be trapped in gate oxide as compared to hot holes while the presence of hot holes generates more interface states. Also, we have demonstrated a new method for extracting the spatial distribution of oxide trapped charge, Q(ot), through gate-to-substrate capacitance (C-gb) measurement. This method is simple to implement and does not require additional information from simulation or detailed knowledge of the device's structure. (C) 2001 Elsevier Science Ltd. All rights reserved.