Dehydromonocrotaline generates sequence-selective N-7 guanine alkylation and heat and alkali stable multiple fragment DNA crosslinks

Monocrotaline is a pyrrolizidine alkaloid known to cause toxicity in humans and animals. Its mechanism of biological action is still unclear although DNA crosslinking has been suggested to a play a role in its activity. In this study we found that an active metabolite of monocrotaline, dehydromonocrotaline (DHM), alkylates guanines at the N7 position of DNA with a preference for 5'-GG and 5'-GA sequences; In addition, it generates piperidine- and heat-resistant multiple DNA crosslinks, as confirmed by electrophoresis and electron microscopy. On the basis of these findings, we propose that DHM undergoes rapid polymerization to a structure which is able to crosslink several fragments of DNA.

Effects of beta-escin and saponin on the transverse-tubular system and sarcoplasmic reticulum membranes of rat and toad skeletal muscle

Mechanically skinned skeletal muscle fibres from rat and toad were exposed to the permeabilizing agents beta-escin and saponin. The effects of these agents on the sealed transverse tubular system (t-system) and sarcoplasmic reticulum (SR) were examined by looking at changes in the magnitude of the force responses to t-system depolarization, the time course of the fluorescence of fura-2 trapped in the sealed t-system, and changes in the magnitude of caffeine-induced contractures following SR loading with Ca2+ under defined conditions. In the presence of 2 mu g ml(-1) beta-escin and saponin, the response to t-system depolarization was not completely abolished, decreasing to a plateau, and a large proportion of fura-2 remained in the sealed t-system. At 10 mu g ml(-1), both agents abolished the ability of both rat and toad preparations to respond to t-system depolarization after 3 min of exposure, but a significant amount of fura-2 remained in sealed t-tubules even after exposure to 100 mu g ml(-1) beta-escin and saponin for 10 min. beta-Escin took longer than saponin to reduce the t-system depolarizations and fura-2 content of the sealed t-system to a similar level. The ability of the SR to load Ca2+ was reduced to a lower level after treatment with beta-escin than saponin. This direct effect on the SR occurred at much lower concentrations for rat (2 mu g ml(-1) beta-escin and 10 mu g ml(-1) saponin) than toad (10 mu g ml(-1) beta-escin and 150 mu g ml(-1) saponin). The reverse order in sensitivities to beta-escin and saponin of t-system and SR membranes indicates that the mechanisms of action of beta-escin and saponin are different in the two types of membrane. In conclusion, this study shows that: (1) beta-escin has a milder action on the surface membrane than saponin; (2) beta-escin is a more potent modifier of SR function; (3) simple permeabilization of membranes is not sufficient to explain the effects of beta-escin and saponin on muscle membranes; and (4) the t-system network within muscle fibres is not a homogeneous compartment.

Effects of medium thermoelasticity on high Rayleigh number steady-state heat transfer and mineralization in deformable fluid-saturated porous media heated from below

In this paper, a solution method is presented to deal with fully coupled problems between medium deformation, pore-fluid flow and heat transfer in fluid-saturated porous media having supercritical Rayleigh numbers. To validate the present solution method, analytical solutions to a benchmark problem are derived for some special cases. After the solution method is validated, a numerical study is carried out to investigate the effects of medium thermoelasticity on high Rayleigh number steady-state heat transfer and mineralization in fluid-saturated media when they are heated from below. The related numerical results have demonstrated that: (1) medium thermoelasticity has a little influence on the overall pattern of convective pore-fluid flow, but it has a considerable effect on the localization of medium deformation, pore-fluid flow, heat transfer and mineralization in a porous medium, especially when the porous medium is comprised of soft rock masses; (2) convective pore-fluid flow plays a very important role in the localization of medium deformation, heat transfer and mineralization in a porous medium. (C) 1999 Elsevier Science S.A. All rights reserved.

Turn up the HEAT

The recently determined crystal structure of the PR65/A subunit of protein phosphatase 2A reveals the architecture of proteins containing HEAT repeats, The structural properties of this solenoid protein explain many functional characteristics and account for the involvement of solenoids as scaffold, anchoring and adaptor proteins.

Whole-body pre-cooling and heat storage during self-paced cycling performance in warm humid conditions

The aim of this study was to establish the effect that pre-cooling the skin without a concomitant reduction in core temperature has on subsequent self-paced cycling performance under warm humid (31 degrees C and 60% relative humidity) conditions. Seven moderately trained males performed a 30 min self-paced cycling trial on two separate occasions. The conditions were counterbalanced as control or whole-body pre-cooling by water immersion so that resting skin temperature was reduced by approximate to 5-6 degrees C. After pre-cooling, mean skin temperature was lower throughout exercise and rectal temperature was lower (P < 0.05) between 15 and 25 min of exercise. Consequently, heat storage increased (P < 0.003) from 84.0 +/- 8.8 W . m(-2) to 153 +/- 13.1 W . m(-2) (mean +/- s((x) over bar)) after pre-cooling, while total body sweat fell from 1.7 +/- 0.1 1 . h(-1) to 1.2 +/- 0.1 1 . h(-1) (P < 0.05). The distance cycled increased from 14.9 +/- 0.8 to 15.8 +/- 0.7 km (P < 0.05) after pre-cooling. The results indicate that skin pre-cooling in the absence of a reduced rectal temperature is effective in reducing thermal strain and increasing the distance cycled in 30 min under warm humid conditions.

Changes in quantum efficiency of Photosystem II of symbiotic dinoflagellates of corals after heat stress, and of bleached corals sampled after the 1998 Great Barrier Reef mass bleaching event Ross J. Jones , Selina Ward, Affendi Yang Amri and Ove Hoegh-Guldberg

Pulse-amplitude-modulation chlorophyll fluorometry was used to examine changes in dark-adapted F-v/F-m of endosymbiotic dinoflagellate microalgae within the tissues of the temperate coral Plesiastrea versipora exposed to elevated seawater temperature. The F-v/F-m was markedly reduced following exposure of corals to 28 degrees C for 48 h. When corals were returned to ambient (24 degrees C) conditions, F-v/F-m increased in an initial rapid and then secondary slower phase. Tissue discolouration (coral bleaching), caused by a significant decrease in the density of algae, was observed during the first 2-3 days of the recovery period. After 14 days, F-v/F-m was still significantly lower than in control corals. The recovery of F-v/F-m is discussed in terms of repair processes within the symbiotic algae, division of healthy algae and also the selective removal of photo-damaged dinoflagellates. Under field conditions, bleached corals sampled at Heron Island Reef during a bleaching event had significantly lower F-v/F-m than non-bleached colonies; four months after the bleaching event, there were no differences in F-v/F-m or algal density in corals marked as having bleached or having shown no signs of colour loss. The results of this laboratory and field study are consistent with the hypothesis that an impairment of photosynthesis occurs during heat-stress, and is the underlying cause of coral bleaching.

Finite element analysis of heat transfer and mineralization in layered hydrothermal systems with upward throughflow

We use the finite element method to solve the coupled problem between convective pore-fluid flow, heat transfer and mineralization in layered hydrothermal systems with upward throughflow. In particular, we present the improved rock alteration index (IRAI) concept for predicting the most probable precipitation and dissolution regions of gold (Au) minerals in the systems. To validate the numerical method used in the computation, analytical solutions to a benchmark problem have been derived. After the numerical method is validated, it is used to investigate the pattern of pore-fluid Aom, the distribution of temperature and the mineralization pattern of gold minerals in a layered hydrothermal system with upward throughflow. The related numerical results have demonstrated that the present concept of IRAI is useful and applicable for predicting the most probable precipitation and dissolution regions of gold (Au) minerals in hydrothermal systems. (C) 2000 Elsevier Science S.A. All rights reserved.

Heat flow for Yang-Mills-Higgs fields, part I

The Yang-Mills-Higgs field generalizes the Yang-Mills field. The authors establish the local existence and uniqueness of the weak solution to the heat flow for the Yang-Mills-Higgs field in a vector bundle over a compact Riemannian 4-manifold, and show that the weak solution is gauge-equivalent to a smooth solution and there are at most finite singularities at the maximum existing time.

Human cyclophilin 40 is a heat shock protein that exhibits altered intracellular localization following heat shock

The unactivated steroid receptors are chaperoned into a conformation that is optimal for binding hormone by a number of heat shock proteins, including Hsp90, Hsp70, Hsp40, and the immunophilin, FKBP52 (Hsp56). Together with its partner cochaperones, cyclophilin 40 (CyP40) and FKBP51, FKBP52 belongs to a distinct group of structurally related immunophilins that modulate steroid receptor function through their association with Hsp90. Due to the structural similarity between the component immunophilins, FKBP52 and cyclophilin 40, we decided to investigate whether CyP40 is also a heat shock protein. Exposure of MCF-7 breast cancer cells to elevated temperatures (42 degreesC for 3 hours) resulted in a 75-fold increase in CyP40 mRNA levels, but no corresponding increase in CyP40 protein expression, even after 7 hours of heat stress. The use of cycloheximide to inhibit protein synthesis revealed that in comparison to MCF-7 cells cultured at 37 degreesC, those exposed to heat stress (42 degreesC for 3 hours) displayed an elevated rate of degradation of both CyP40 and FKBP52 proteins. Concomitantly, the half-life of the CyP40 protein was reduced from more than 24 hours to just over 8 hours following heat shock. As no alteration in CyP40 protein levels occurred in cells exposed to heat shock, an elevated rate of degradation would imply that CyP40 protein was synthesized at an increased rate. hence the designation of human CyP40 as a heat shock protein. Application of heat stress elicited a marked redistribution of CyP40 protein in MCF-7 cells from a predominantly nucleolar localization, with some nuclear and cytoplasmic staining, to a pattern characterized by a pronounced nuclear accumulation of CyP40, with no distinguishable nucleolar staining. This increase in nuclear CyP40 possibly resulted from a redistribution of cytoplasmic and nucleolar CyP40, as no net increase in CyP40 expression levels occurred in response to stress. Exposure of MCF-7 cells to actinomycin D for 4 hours resulted in the translocation of the nucleolar marker protein, B23, from the nucleolus, with only a small reduction in nucleolar CyP40 levels. Under normal growth conditions, MCF-7 cells exhibited an apparent colocalization of CyP40 and FKBP52 within the nucleolus.

Probability of axillary node involvement in patients with tubular carcinoma of the breast

The aim of this study was to investigate the frequency of axillary metastasis in women with tubular carcinoma (TC) of the breast. Women who underwent axillary dissection for TC in the Western Sydney area (1984-1995) were identified retrospectively through a search of computerized records. A centralized pathology review was performed and tumours were classified as pure tubular (22) or mixed tubular (nine), on the basis of the invasive component containing 90 per cent or more, or 75-90 per cent tubule formation respectively. A Medline search of the literature was undertaken to compile a collective series (20 studies with a total of 680 patients) to address the frequency of nodal involvement in TC. A quantitative meta-analysis was used to combine the results of these studies. The overall frequency of nodal metastasis was five of 31 (16 per cent); one of 22 pure tubular and four of nine mixed tumours (P = 0.019). None of the tumours with a diameter of 10 mm or less (n = 16) had nodal metastasis compared with five of 15 larger tumours (P = 0.018). The meta-analysis of 680 women showed an overall frequency of nodal metastasis in TC of 13.8 (95 per cent confidence interval 9.3-18.3) per cent. The frequency of nodal involvement was 6.6 (1.7-11.4) per cent in pure TC (n = 244) and 25.0 (12.5-37.6) per cent in mixed TC (n = 149). A case may be made for observing the clinically negative axilla in women with a small TC (10 mm or less in diameter).

Heat flow for the Yang-Mills-Higgs field and the Hermitian Yang-Mills-Higgs metric

For a parameter lambda > 0, we study a type of vortex equations, which generalize the well-known Hermitian-Einstein equation, for a connection A and a section phi of a holomorphic vector bundle E over a Kahler manifold X. We establish a global existence of smooth solutions to heat flow for a self-dual Yang-Mills-Higgs field on E. Assuming the lambda -stability of (E, phi), we prove the existence of the Hermitian Yang-Mills-Higgs metric on the holomorphic bundle E by studying the limiting behaviour of the gauge flow.

Modeling of heat transfer in fluidized-bed coating of cylinders

A numerical model of heat transfer in fluidized-bed coating of solid cylinders is presented. By defining suitable dimensionless parameters, the governing equations and its associated initial and boundary conditions are discretized using the method of orthogonal collocation and the resulting ordinary differential equations simultaneously solved for the dimensionless coating thickness and wall temperatures. Parametric Studies showed that the dimensionless coating thickness and wall temperature depend on the relative heat capacities of the polymer powder and object, the latent heat of fusion and the size of the cylinder. Model predictions for the coating thickness and wall temperature compare reasonably well with numerical predictions and experimental coating data in the literature and with our own coating experiments using copper cylinders immersed in nylon-11 and polyethylene powders. (C) 2001 Elsevier Science Ltd. All rights reserved.

Finite element modelling of heat transfer through permeable cracks in hydrothermal systems with upward throughflow

We use the finite element method to model the heat transfer phenomenon through permeable cracks in hydrothermal systems with upward throughflow. Since the finite element method is an approximate numerical method, the method must be validated before it is used to soh,e any new, kind of problem. However, the analytical solution, which can be used to validate the finite element method and other numerical methods, is rather limited in the literature, especially, for the problem considered here. Keeping this in mind, we have derived analytical solutions for the temperature distribution along the vertical axis of a crack in a fluid-saturated porous layer. After the finite element method is validated by comparing the numerical solution with the analytical solution for the same benchmark problem, it is used to investigate the pore-fluid flow and heat transfer in layered hydrothermal systems with vertical permeable cracks. The related analytical and numerical results have demonstrated that vertical cracks are effective and efficient members to transfer heat energy from the bottom section to the top section in hydrothermal systems with upward throughflow.

Tubular system volume changes in twitch fibres from toad and rat skeletal muscle assessed by confocal microscopy

The volume of the extracellular compartment (tubular system) within intact muscle fibres from cane toad and rat was measured under various conditions using confocal microscopy. Under physiological conditions at rest, the fractional volume of the tubular system (t-sys(Vol)) was 1.38 +/- 0.09% (n = 17),1.41 +/- 0.09% (n = 12) and 0.83 +/- 0.07% (n = 12) of the total fibre volume in the twitch fibres from toad iliofibularis muscle, rat extensor digitorum longus muscle and rat soleus muscle, respectively. In toad muscle fibres, the t-sys(Vol) decreased by 30% when the tubular system was fully depolarized and decreased by 15% when membrane cholesterol was depleted from the tubular system with methyl-beta-cyclodextrin but did not change as the sarcomere length was changed from 1.93 to 3.30 mum. There was also an increase by 30% and a decrease by 25% in t-sys(Vol) when toad fibres were equilibrated in solutions that were 2.5-fold hypertonic and 50% hypotonic, respectively. When the changes in total fibre volume were taken into consideration, the t-sys(Vol) expressed as a percentage of the isotonic fibre volume did actually decrease as tonicity increased, revealing that the tubular system in intact fibres cannot be compressed below 0.9% of the isotonic fibre volume. The results can be explained in terms of forces acting at the level of the tubular wall. These observations have important physiological implications showing that the tubular system is a dynamic membrane structure capable of changing its volume in response to the membrane potential, cholesterol depletion and osmotic stress but not when the sarcomere length is changed in resting muscle.