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Paclitaxel prevents loss of pulmonary endothelial barrier integrity during cold preservation.

BACKGROUND: Cold preservation is the most practical method to maintain the viability of isolated lungs. However, rapid cooling may affect pulmonary endothelial function. We examined the effects of microtubule stabilization with paclitaxel on pulmonary endothelial barrier integrity under cold temperature. METHODS: Human pulmonary arterial endothelial cells were incubated at 4 degrees C for 2 hr in the presence or absence of paclitaxel (2.5 micromol/L). Microtubules was visualized using immunocytochemical techniques. Ultrasonic attenuation was measured with scanning acoustic microscopy. Endothelial barrier integrity was measured as transendothelial electric resistance. In addition, we examined graft function in a rat lung transplantation model, in which the donor lung had been preserved in the presence of paclitaxel (2.5 micromol/L) at 4 degrees C for 12 hr. RESULTS: Low temperature caused a reversible microtubule disassembly, but the structure of microtubules was preserved by paclitaxel. Paclitaxel prevented the cooling-induced decrease in ultrasonic attenuation and transendothelial electric resistance. In a rat transplantation model, we found that preservation with paclitaxel successfully improved the oxygenation performance of the donor lung, which demonstrated only mild congestion and less significant interstitial edema without fluid accumulation in the alveolar spaces. CONCLUSIONS: Our results indicate that microtubule stabilization with paclitaxel may be beneficial to prevent the loss of the endothelial barrier during cold preservation. We conclude that the use of paclitaxel in organ preservation solutions is useful in protecting pulmonary endothelial barrier integrity during cold preservation, thereby reducing the occurrence of early graft failure.

Animals↗

Northern elephant seal platelets: analysis of shape change and response to platelet agonists.

Blood platelets have a vital role in the maintenance of normal mammalian hemostasis. Rapid pressure changes and temperatures lower than 20 degrees C cause activation of human and terrestrial mammal platelets. Elephant seals are routinely subjected to pressures as high as 150 atm, yet do not suffer from the thrombotic effects of platelet activation associated with rapid decompression. We examined the ultrastructure of Northern elephant seal (Mirounga angustirostris) platelets and their functional and morphological response to various platelet agonists. Unstimulated elephant seal platelets are discoid cells, with a microtubule coil, randomly dispersed alpha and dense granules, and glycogen granules. There are well-defined areas of membranous invaginations that indicate the presence of an open canalicular system (OCS). Aggregometry was used to determine the response of elephant seal platelets to various platelet agonists. Dose-dependent curves were generated for thrombin, collagen, and adenosine diphosphate (ADP). Platelet response to thrombin was dose-dependent and was maximal at 2.5 U/ml. Platelets collected in sodium citrate had blunted responses to both ADP and collagen. ADP stimulation caused only reversible, primary activation (shape change) at > or = 5 microM, while platelets did not aggregate in response to any concentration of collagen. Platelets collected in sodium heparin did respond fully to both to ADP and collagen. There was small, reversible shape change in response to ristocetin, but no response to epinephrine. Decreased sensitivity of elephant seal platelets to agonists may be a protective mechanism developed in response to rapid pressure changes and cold temperatures associated with adaptation to an extreme environment.

Adaptation, Physiological↗

Spatiotemporal analysis of environmental exposure-health effect associations.

The goal of this work is to discuss a general methodology for studying associations between environmental exposures and health effect by means of the spatiotemporal random field theory. This theory is the tool of choice for rigorously accounting for important spatiotemporal variations and uncertainties related to exposures and effect. Within the framework of the random field theory, the Bayesian maximum entropy model neatly synthesizes various sources of physical and epidemiological knowledge into spatiotemporal analysis. Therefore, unlike technical statistics, this approach relies on the blending of substantive physical knowledge with powerful mathematical techniques and a coherent rationale. Given the well-founded fact that certain health effects may be caused by environmental exposures, the significance of these exposures is assessed in terms of a criterion that is based on the joint stochastic representation of exposure and health-effect distributions in space/time. In view of this criterion, the strength and consistency of the exposure-effect association are evaluated on the basis of the health-effect predictions that the combined physico-epidemiologic analysis generates in space/time. The main features of the approach are demonstrated by a simulation example and a real case study involving mortality and cold temperatures in North Carolina. The studies demonstrated the practical usefulness of the stochastic human exposure analysis in assessing the exposure-effect association. The results reported here emphasize the links between spatiotemporal models of physical systems and population health-effect distributions, thus suggesting directions for improving the current understanding of quantitative "exposure-health effect" functions.

Cold Temperature↗

Energetics of nestling growth and parental effort in Antarctic fulmarine petrels.

Antarctic fulmarine petrels breed in some of the coldest conditions encountered by any bird and their young grow twice as fast as predicted allometrically. To examine the energetic consequences of fast growth in a cold environment, we used the doubly labeled water technique to measure field metabolic rates of adults (three species) and different-aged nestlings (four species) of Antarctic fulmarine petrels in the Rauer Islands, East Antarctica: Antarctic fulmar Fulmarus glacialoides, Antarctic petrel Thalassoica antarctica, Cape petrel Daption capense and snow petrel Pagodroma nivea. We used our data to assess parental effort and, together with literature values on nestling growth and resting metabolic rate, to construct and partition nestling energy budgets. Nestling total energy expenditure and peak daily metabolic rate were significantly higher than predicted allometrically (33-73% and 17-66% higher, respectively), and the relative cost of growth in nestling petrels was among the highest reported for birds (54-72 kJ g(-1)). Parental effort during the nestling-feeding period was identical in adult Cape and Antarctic petrels (3.5 times basal metabolic rate, BMR), and was somewhat (but not significantly) higher in snow petrels (4.6 times BMR). These values are comparable to those of other high-latitude procellariiform birds. Thus, despite the constraints of a compressed breeding season, cold temperatures and fast-growing nestlings, adult Antarctic fulmarine petrels do not work harder than procellariid adults whose chicks grow much more slowly. Our findings suggest that obtaining sufficient food is generally not a constraint for adult fulmarine petrels and that factors operating at the tissue level limit nestling growth rate.

Adaptation, Physiological↗

Economic change and sex-specific cardiovascular mortality in Britain 1955-1976.

Analyses directed toward recent declines in cardiovascular disease mortality rates have typically focused on alterations in important physiological and behavioral risk factors resulting from lifestyle changes and medical advances. In this study, a multivariate model of the impact of more fundamental changes in the socioeconomic and bio-physical environments has been developed and applied to cardiovascular disease mortality rates, by sex, in England and Wales and Scotland during 1955-1976. The predictive model includes factors associated with (1) long-term growth in the economy, (2) deleterious behavioral risk factors loosely associated with economic growth--especially cigarette consumption per capita, (3) economic instability--especially recession as indicated by factors related to unemployment, income loss, and recessional declines in average weekly hours worked in manufacturing industries, (4) health care, and (5) physical environmental disturbances--especially very cold temperatures. This model proves to be an excellent instrument for the statistical explanation of trends and fluctuations in CVD mortality rates for both sexes and both regions in Britain in the post-War period. In general, the overall exponential rate of economic growth is found to be the most powerful factor in the long-term decline in CVD mortality rates. Similarly, disturbances to the national and regional economic situations--especially recessions--have regularly been associated with elevated death rates for all populations observed. Cigarette and unusually high spirits consumption, as well as particularly cold winter temperatures, have also had important deleterious effects on CVD mortality. The proportion of government expenditures devoted to health care is associated with a reduction in CVD mortality in England and Wales.

Adult↗

Cryoactivation and tryptic activation of blood 'prorenin' in normal man and animals.

Plasma and serum of healthy subjects apparently contain a precursor form of renin, or 'prorenin,' which can be activated by the ice-cold temperature at which samples are customarily handled for prolonged periods in laboratories and blood banks. The effect of such prior cryoactivation for 9 days at 4 degrees C is to increase subsequent plasma renin activity (PRA) at 37 degrees C by 108 +/- 16.3% (mean +/- SE) over the nonactivated control value (P less than 0.001). At a lower temperature (-4 degrees C), the cryoactivation effect is considerably greater than at 4 degrees C. Cryoactivation is not obliterated by the prefreezing of plasma, or reduced by inclusion of bacteriostats. Nor is it attributable to any detectable reduction in angiotensinase activity. In rats, cryoactivation at 4 degrees C is much lower than in humans, suggesting a marked species difference either in prorenin concentration or in the rapidity of its spontaneous conversion after blood collection. Trypsin at near optimal concentrations also consistently activates human plasma prorenin, whether at 4, 23, or 37 degrees C indicating that cold is not an essential concomitant of tryptic activation. In excess, the magnitude of which varies among individuals, trypsin at first produces activation and later a decline in PRA, probably due to degradation of the reactants (prorenin, renin, angiotensinogen) and of the initial product (angiotensin I). The identity of angiotensin I in activated and control plasmas can be established by specific radioimmunoassay, and bioassay. Our data indicate that tryptic activation involves little direct production of angiotensin I but rather converts prorenin, thereby enhancing the angiotensin generating capacity of the plasma renin system itself. Tryptic activation in plasma of anaesthetized dogs is lower than in humans, but higher than in conscious or anaesthetized rabbits in whom the effect appears to be slight. In anaesthetized rats there is virtually no tryptic activation, which is in line with the results by cryoactivation. Since the renin--angiotensin systems of dogs, rabbits, and rats have been extensively studied in experimental models of human hypertension, these observed departures from human levels of cryoactivation and tryptic activation of prorenin deserve further investigation.

Adult↗

Effect of chronic cold and submergence on blood oxygen transport in the turtle, Chrysemys picta.

Whole blood oxygen equilibrium curves (O2EC's) and related hematologic properties are reported for the turtle Chrysemys picta exposed to two experimental conditions. Summer turtles were maintained at 24 degrees C with free access to air; winter turtles were submerged for 4-12 wk in N2-bubbled water at 3 degrees C. Half-saturation Po2's at 3 degrees C for blood from summer and winter animals were 4.1 and 4.5 Torr, respectively. At 24 degrees C, summer and winter P50's were 20.2 and 22.7 Torr, respectively. The winter turtle P50 values were lower than predicted since prolonged submergence effected a severe metabolic acidosis; blood pH's for winter turtles were 0.65 pH unit lower than for summer animals at both temperatures. Cold submergence also had a profound influence on O2EC shape. Winter turtle curves exhibited high O2 affinity below P50 while they were distinctly right-shifted above 50% S. Winter animals also exhibited reduced CO2-Bohr coefficients (delta log Po2/delta pH) at 3 and 24 degrees C. Prolonged submergence did not affect the animal's isohemoglobin profile (demonstrated by isoelectric focusing) or [metHb]. The [ATP] and [DPG] in winter turtle red cells, however, decreased significantly; the ratio of organic phosphate ([ATP] + [DPG]) to Hb tetramer fell from 1.4 in summer animals to 0.5 in winter turtles. These findings suggest that the effect of chronic cold and prolonged submergence on turtle O2EC position and shape may result from reduction in RBC organic phosphates. Furthermore, these observed changes in blood oxygen transport may facilitate O2 loading during winter submergence via extrapulmonary gas exchange.

Acid-Base Equilibrium↗

The need for winter in the switch to flowering.

Vernalization is the process whereby the floral transition is promoted through exposure of plants to long periods of cold temperature or winter. A requirement for vernalization aligns flowering with the seasons to ensure that their reproductive phase occurs in favorable conditions. The mitotic stability of vernalization, suggestive of an epigenetic mechanism, has intrigued researchers for many years. Genetic analysis of the vernalization requirement in Arabidopsis has identified key floral repressor genes, FRI and FLC. The action of these floral repressors is antagonized by vernalization and the activity of a set of genes grouped into the autonomous floral pathway. Analysis of the vernalization pathway has defined a series of epigenetic regulators crucial for "cellular-memory" of the cold signal, whereas the autonomous pathway appears to function in part through posttranscriptional mechanisms. The mechanism of the vernalization requirement, which is now being explored in a range of plant species, should uncover the evolutionary origins of this key agronomic trait.

Arabidopsis↗

[Microbial diversity of deep-sea extremophiles--Piezophiles, Hyperthermophiles, and subsurface microorganisms].

Knowledge of our Planet's biosphere has increased tremendously during the last 10 to 20 years. In the field of Microbiology in particular, scientists have discovered novel "extremophiles", microorganisms capable of living in extreme environments such as highly acidic or alkaline conditions, at high salt concentration, with no oxygen, extreme temperatures (as low as -20 degrees C and as high as 300 degrees C), at high concentrations of heavy metals and in high pressure environments such as the deep-sea. It is apparent that microorganisms can exist in any extreme environment of the Earth, yet already scientists have started to look for life on other planets; the so-called "Exobiology" project. But as yet we have little knowledge of the deep-sea and subsurface biosphere of our own planet. We believe that we should elucidate the Biodiversity of Earth more thoroughly before exploring life on other planets, and these attempts would provide deeper insight into clarifying the existence of extraterrestrial life. We focused on two deep-sea extremophiles in this article; one is "Piezophiles", and another is "Hyperthermophiles". Piezophiles are typical microorganisms adapted to high-pressure and cold temperature environments, and located in deep-sea bottom. Otherwise, hyperthermophiles are living in high temperature environment, and located at around the hydrothermal vent systems in deep-sea. They are not typical deep-sea microorganisms, but they can grow well at high-pressure condition, just like piezophiles. Deming and Baross mentioned that most of the hyperthermophilic archaea isolated from deep-sea hydrothermal vents are able to grow under conditions of high temperature and pressure, and in most cases their optimal pressure for growth was greater than the environmental pressure they were isolated from. It is possible that originally their native environment may have been deeper than the sea floor and that there had to be a deeper biosphere. This implication suggests that the deep-sea hydrothermal vents are the windows to a deep subsurface biosphere. A vast array of chemoautotrophic deep-sea animal communities have been found to exist in cold seep environments, and most of these animals are common with those found in hydrothermal vent environments. Thus, it is possible to consider that the cold seeps are also one of slit windows to a deep subsurface biosphere. We conclude that the deep-sea extremophiles are very closely related into the unseen majority in subsurface biosphere, and the subsurface biosphere probably concerns to consider the "exobiology".

Archaea↗

Studies on calcium regulation in relation to sodium-potassium balance in the rabbit lens.

Net changes in the levels of lens calcium, magnesium, sodium, potassium, and water content were determined following exposure in vitro to iodoacetate, ouabain, lanthanum, cyanide, quercetin and cold temperature. The lens calcium content rose following inhibition of lens metabolism but did not change following specific inhibition of the sodium pump. The data revealed that regulation of the lens calcium balance is largely independent of lens sodium and potassium content. A greater increase in lens sodium content upon iodoacetate treatment vs. ouabain treatment is discussed in relation to the elevation of lens calcium.

Animals↗

Effect of temperature on apical membrane remodeling in ADH-stimulated toad urinary bladders.

Pretreatment and removal of vasopressin (ADH) in toad urinary bladder renal model tissues induces endocytosis at 25 degrees C. The objective of the current study is to determine if apical membrane remodeling, as well as transepithelial water flow, can be affected by lowering the temperature to 15 degrees C. Control toad urinary bladders in the presence of an osmotic gradient at either 25 degrees C or 15 degrees C when visualized by scanning electron microscopy (SEM) show a typical apical membrane surface with no apparent surface differences. ADH-treated tissues following 15-min stimulation at 25 degrees C or 15 degrees C revealed a propagation of apical microvilli on their surface membranes. After 15 min following removal of ADH, bladder tissues at 25 degrees C or 15 degrees C showed surface invaginations involving over 44% and 80% of granular cells, respectively. The rate of water flow in tissues at 15 degrees C remained elevated compared to tissues held at 25 degrees C. This was consistent with the observation that ADH-stimulated tissues following washout at 15 degrees C still had marked apical membrane surface involvement. However, at 30 min and 60 min postwashout, ADH-stimulated tissues at 15 degrees C recovered considerably, with a reduction in the number of shallow apical membrane invaginations involving fewer than 33% and 20% of granular cells respectively. This may indicate that the membrane undergoes continuous remodeling even at cold temperature conditions but with a different half-time. Control bladder tissues subjected to transmission electron microscopy (TEM) reveal a dense cytoplasmic profile with a scattered distribution of secretory granules, rough ER cisternae, mitochondria, and little or no vacuolation. In contrast, ADH-stimulated bladder tissues displayed a vacuolated cytoplasm, expanded rough ER cisternae, and ruffled basolateral membranes. These observations suggest that the apical membrane undergoes considerable reorganization following cessation of hormone action and that lowering the temperature reduces the rate of membrane remodeling and thus may provide a means to monitor the processes of endocytosis and the mechanisms responsible for water channel retrieval.

Animals↗

Cold-induced impairment of delayed matching in rats.

Exposure to moderate, nonhypothermic cold temperature has been reported to affect a variety of behavioral and neural functions. To elucidate the effects of mild cold stress on short-term (working) memory, Long-Evans rats were exposed to an ambient temperature of either 2 degrees or 23 degrees C while performing a delayed matching task. At the beginning of each trial, rats were required to respond on one of two levers cued by a light. Following a delay of 2, 8, or 16 s, a response on the lever previously cued produced food reinforcement. Relative to performance at 23 degrees C, exposure to 2 degrees C occasioned no change in matching accuracy at the 2-s delay, a modest decrement at the 8-s delay, and a larger decrement at the 16-s delay. The cold exposure did not decrease colonic temperature. In addition to accuracy decrements, matching response times were consistently shorter during cold exposures. Cold-induced impairments were absent during removal of the memory component from the task, indicating the observed cold effects on memory were not due to impaired attentional, sensory, or motor processes. These data suggest that mild cold stress may impair active maintenance of information in working memory but not processes related to reference memory.

Animals↗

Hormonal responses to exercise during moderate cold exposure in young vs. middle-age subjects.

The influence of moderate cold exposure on the hormonal responses of atrial natriuretic factor (ANF), arginine vasopressin (AVP), catecholamines, and plasma renin activity (PRA) after exhaustive exercise was studied in 9 young and 10 middle-aged subjects. Exercise tests were randomly performed in temperate (30 degrees C) and cold (10 degrees C) environments. Heart rate, oxygen consumption, and peripheral arterial blood pressure were measured at regular intervals. Blood samples were collected before and immediately after exercise at 30 or 10 degrees C. Plasma sodium and potassium concentrations as well as hemoglobin and hematocrit were measured, and the change in plasma volume was calculated. At rest and during exercise, oxygen consumption was similar during exposure to both temperate and cold temperatures. During submaximal exercise intensities, the rise in heart rate was blunted while the increase in systolic blood pressure was significantly greater at 10 than at 30 degrees C. The increases in plasma sodium and potassium concentrations after exhaustion were similar between environments, as was the decrease in plasma volume. In both groups, all plasma hormones were significantly elevated postexercise, with the AVP response similar at 10 and 30 degrees C. However, the norepinephrine and ANF responses were significantly greater while the PRA response was significantly reduced at 10 degrees C. In the middle-aged subjects the epinephrine response to exercise was higher at 10 than at 30 degrees C. The greater ANF and reduced PRA responses to exercise in the cold may have resulted from central hemodynamic changes caused by cold-induced cutaneous vasoconstriction.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

It's time to flower: the genetic control of flowering time.

In plants, successful sexual reproduction and the ensuing development of seeds and fruits depend on flowering at the right time. This involves coordinating flowering with the appropriate season and with the developmental history of the plant. Genetic and molecular analysis in the small cruciform weed, Arabidopsis, has revealed distinct but linked pathways that are responsible for detecting the major seasonal cues of day length and cold temperature, as well as other local environmental and internal signals. The balance of signals from these pathways is integrated by a common set of genes to determine when flowering occurs. Excitingly, it has been discovered that many of these same genes regulate flowering in other plants, such as rice. This review focuses on recent advances in how three of the signalling pathways (the day-length, vernalisation and autonomous pathways) function to control flowering.

Arabidopsis↗

Escherichia coli aminodeoxychorismate synthase: analysis of pabB mutations affecting catalysis and subunit association.

p-Aminobenzoic acid (PABA), an essential component of the vitamin folic acid, is derived from the aromatic branch-point precursor chorismate in two steps. 4-Amino-4-deoxychorismate (ADC) synthase converts chorismate and glutamine to ADC and glutamate, and is composed of two subunits, PabA and PabB. While various experiments have suggested that PabA and PabB act as a complex, attempts to isolate the intact complex have failed. We report here the first successful copurification of PabA and PabB by gel filtration chromatography. The association of PabA and PabB is greatly enhanced by the presence of 5 mM glutamine, and by preincubation at 37 degrees C. Conversely, the association is greatly reduced at cold temperatures. We also report the isolation and characterization of both chemically induced and site-directed mutations in PabB. Mutated PabB enzymes fall into three categories according to their properties: deficiency of chorismate amination coupled with failure to associate with PabA, deficiency of chorismate amination coupled with retention of PabA association, and competency of chorismate amination with failure of PabA association.

Amination↗

[Diseases caused by repetitive trauma of the upper limbs: epidemiology, diagnosis, prevention. Epidemiological aspects].

The association between occupational risk factors and musculo-skeletal disorders due to biomechanical overload (WMSDs) has been focused on numerous research projects, ranging from those simple observing the different pathological findings reported among workers performing particular tasks, down to the latest studies actually quantifying the exposure of workers to basic risk factors, as force, awkward posture, repetition and to modifying factors as duration, intensity, temporal profile and cold temperatures. The critical review of the epidemiological studies, taking into account their quality, showed a clear relationship between basic and modifying risk factors and upper limb pathologies. In particular, force exertion has been shown strongly related to neck, elbow and hand-wrist pathologies; repetition showed some convincing evidence of causal relationship to neck, shoulder and hand-wrist pathologies. Insufficient evidence of work relatedness has been shown between both repetition and awkward posture and elbow pathologies.

Animals↗

Mitochondrial UCP4 mediates an adaptive shift in energy metabolism and increases the resistance of neurons to metabolic and oxidative stress.

The high-metabolic demand of neurons and their reliance on glucose as an energy source places them at risk for dysfunction and death under conditions of metabolic and oxidative stress. Uncoupling proteins (UCPs) are mitochondrial inner membrane proteins implicated in the regulation of mitochondrial membrane potential (Deltapsim) and cellular energy metabolism. The authors cloned UCP4 cDNA from mouse and rat brain, and demonstrate that UCP4 mRNA is expressed abundantly in brain and at particularly high levels in populations of neurons believed to have high-energy requirements. Neural cells with increased levels of UCP4 exhibit decreased Deltapsim, reduced reactive oxygen species (ROS) production and decreased mitochondrial calcium accumulation. UCP4 expressing cells also exhibited changes of oxygen-consumption rate, GDP sensitivity, and response of Deltapsim to oligomycin that were consistent with mitochondrial uncoupling. UCP4 modulates neuronal energy metabolism by increasing glucose uptake and shifting the mode of ATP production from mitochondrial respiration to glycolysis, thereby maintaining cellular ATP levels. The UCP4-mediated shift in energy metabolism reduces ROS production and increases the resistance of neurons to oxidative and mitochondrial stress. Knockdown of UCP4 expression by RNA interference in primary hippocampal neurons results in mitochondrial calcium overload and cell death. UCP4-mRNA expression is increased in neurons exposed to cold temperatures and in brain cells of rats maintained on caloric restriction, suggesting a role for UCP4 in the previously reported antiageing and neuroprotective effects of caloric restriction. By shifting energy metabolism to reduce ROS production and cellular reliance on mitochondrial respiration, UCP4 can protect neurons against oxidative stress and calcium overload.

Adenosine Triphosphate↗

Thermal balance of cattle grazing winter range: model application.

Beef cattle grazing semiarid foothill rangeland of the Northern Rockies during winter may be exposed to cold temperatures and high winds while grazing pastures with low nutritional value. Cattle can physiologically and behaviorally respond to the changing environment to lower their metabolic requirements and reduce the effects of cold exposure. Requirements of grazing cattle may be overpredicted with models developed in controlled settings that do not account for energy-conserving behaviors. We refined a simple thermal balance equation to model heat exchange of free-ranging cattle. We accounted for the complex interactions between animal behavior and the changing natural environment by applying the insulation characteristics of the cattle's tissue and coat to a simple geometric shape of an asymmetric ellipsoid at different orientations to the sun and wind. We compared the model predictions with heat production measured in 3 studies, and in all cases the model predictions were similar to those reported. Model simulations indicate behaviors, such as lying and orientation to the sun, mitigated the effects of extreme weather. For many combinations of winter weather variables, metabolic requirements increased only slightly due to cold exposure of mature beef cattle in a near-maintenance state. The results indicate that solar radiation contributes strongly to the thermal balance of a cow. Thus, previous models that do not account for the irradiative environment may overestimate metabolic requirements of cattle acclimated to grazing winter range.

Acclimatization↗