Chilling findings: the need for winter vigilance in heart failure.
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The land snail Helix pomatia (Gastropoda: Helicidae) is widely distributed in Northern and Central Europe where it may experience subzero temperatures during winter months. Its supercooling ability was studied in two populations of H. pomatia. One population originated from Southern Sweden (Gotaland) and the other from Central France (Auvergne). In the experimental design, they were acclimated, over 2 weeks, to artificial winter conditions (hibernation, T=5 degrees C). The Swedish snails showed a rather limited supercooling ability (temperature of crystallization, T(c)=-6.4+/-0.8 degrees C), significantly greater, however, than the supercooling capacity of the population from France (T(c)=-4.6+/-1.4 degrees C). In artificial spring conditions (3 months of hibernation followed by a progressive acclimation, over 2 weeks, to activity at T=20 degrees C), both populations exhibited a similar high T(c) (-2.0+/-1.0 degrees C). The lower T(c) of hibernating Swedish snails could be due to a greater loss of body water, accompanied by a higher concentration of solutes in the hemolymph. In both populations, the variation in hemolymph osmolality measured between hibernating (250-270 mOsm kg(-1)) and active (165-215 mOsm kg(-1)) snails may be explained by the variation in body water mass and did not suggest the production of colligative cryoprotectants. Moreover, the three bacterial strains, Buttiauxella sp., Kluyvera sp., and Tatumella sp. (Enterobacteriaceae) which were isolated from fed snails, but absent in starved snails, did not show any ice-nucleating activity at temperatures higher than -9 degrees C. Only the strain Kluyvera sp. initiated nucleation at -9 degrees C. This strain, therefore, is a weak, also termed a Type III or Class C ice-nucleating active bacterium, but with no influence on the supercooling ability of individual snails. In summary, fluctuations in body water mass of hibernating snail populations, triggering changes in osmolyte concentration, rather than the presence of endogenous ice-nucleating-active bacteria, accounts for fluctuations in their T(c).
The Antarctic nematode Panagrolaimus davidi has an ice-active protein that shows recrystallization inhibition but no thermal hysteresis. It belongs to a class of ice-active proteins found in a variety of freezing-tolerant organisms that display insignificant levels of thermal hysteresis in the context of the environmental temperatures to which they are exposed. The recrystallization inhibition activity of the P. davidi ice-active protein is present at low concentrations, is relatively heat stable, is affected more by acid than by alkaline pH, is not calcium dependant and is not affected by reagents that target carbohydrate residues or sulphydryl linkages. A hexagonal ice crystal growth form also indicates the presence of an ice-active protein. This protein could have important functions in the survival of intracellular freezing by this organism by controlling the stability of ice after its formation.
Antifreeze proteins are characterised by their ability to prevent ice from growing upon cooling below the bulk melting point. This displacement of the freezing temperature of ice is limited and at a sufficiently low temperature a rapid ice growth takes place. The separation of the melting and freezing temperature is usually referred to as thermal hysteresis, and the temperature of ice growth is referred to as the hysteresis freezing point. The hysteresis is supposed to be the result of an adsorption of antifreeze proteins to the crystal surface. This causes the ice to grow as convex surface regions between adjacent adsorbed antifreeze proteins, thus lowering the temperature at which the crystal can visibly expand. The model requires that the antifreeze proteins are irreversibly adsorbed onto the ice surface within the hysteresis gap. This presupposition is apparently in conflict with several characteristic features of the phenomenon; the absence of superheating of ice in the presence of antifreeze proteins, the dependence of the hysteresis activity on the concentration of antifreeze proteins and the different capacities of different types of antifreeze proteins to cause thermal hysteresis at equimolar concentrations. In addition, there are structural obstacles that apparently would preclude irreversible adsorption of the antifreeze proteins to the ice surface; the bond strength necessary for irreversible adsorption and the absence of a clearly defined surface to which the antifreeze proteins may adsorb. This article deals with these apparent conflicts between the prevailing theory and the empirical observations. We first review the mechanism of thermal hysteresis with some modifications: we explain the hysteresis as a result of vapour pressure equilibrium between the ice surface and the ambient fluid fraction within the hysteresis gap due to a pressure build-up within the convex growth zones, and the ice growth as the result of an ice surface nucleation event at the hysteresis freezing point. We then go on to summarise the empirical data to show that the dependence of the hysteresis on the concentration of antifreeze proteins arises from an equilibrium exchange of antifreeze proteins between ice and solution at the melting point. This reversible association between antifreeze proteins and the ice is followed by an irreversible adsorption of the antifreeze proteins onto a newly formed crystal plane when the temperature is lowered below the melting point. The formation of the crystal plane is due to a solidification of the interfacial region, and the necessary bond strength is provided by the protein "freezing" to the surface. In essence: the antifreeze proteins are "melted off" the ice at the bulk melting point and "freeze" to the ice as the temperature is reduced to subfreezing temperatures. We explain the different hysteresis activities caused by different types of antifreeze proteins at equimolar concentrations as a consequence of their solubility features during the phase of reversible association between the proteins and the ice, i.e., at the melting point; a low water solubility results in a large fraction of the proteins being associated with the ice at the melting point. This leads to a greater density of irreversibly adsorbed antifreeze proteins at the ice surface when the temperature drops, and thus to a greater hysteresis activity. Reference is also made to observations on insect antifreeze proteins to emphasise the general validity of this approach.
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Given the unique thermal history of the Antarctic continent, fishes of dominant suborder Notothenioidei offer a remarkable opportunity to study the physiological and biochemical characters gained and, conversely, lost during their evolutionary history and to map this information on the species phylogenetic trees. The availability of phylogenetically related notothenioid taxa living in a wide range of latitudes (in the Antarctic, sub-Antarctic and temperate regions) allows to look into the molecular bases of environmentally driven gene birth and death. This evolutionary perspective has also been supported by comparison of some features of the hemoprotein devoted to the oxygen transport in fish species living in the other polar region, the Arctic. The Arctic and Antarctic marine ichthyofaunas differ by age and isolation. Fishes of the two polar regions have undergone different regional histories which have engineered the physiological diversities, so that Antarctic fish are much more stenothermal than Arctic ones. Understanding the mechanisms of phenotypic response to cold exposure in species living at different latitudes in polar habitats offers fundamental insights into environmental adaptations. This review aims at surveying the current knowledge of molecular structure, functional features, phylogeny and adaptations of the hemoglobin of fish thriving in the Antarctic, sub-Antarctic and Arctic regions (with some excursions in the temperate latitudes). Investigating the evolutionary adaptations of hemoglobins to these environments can provide new insights into adaptation currently studied merely in temperate organisms, and can shed light into the convergent processes that evolved in response to thermal adaptations.
In situ biodegradation of benzene, toluene, and xylenes in a petroleum hydrocarbon contaminated aquifer near Fairbanks, Alaska was assessed using carbon and hydrogen compound specific isotope analysis (CSIA) of benzene and toluene and analysis of signature metabolites for toluene (benzylsuccinate) and xylenes (methylbenzylsuccinates). Carbon and hydrogen isotope ratios of benzene were between -25.9 per thousand and -26.8 per thousand for delta13C and -119 per thousand and -136 per thousand for delta2H, suggesting that biodegradation of benzene is unlikely at this site. However, biodegradation of both xylenes and toluene were documented in this subarctic aquifer. Biodegradation of xylenes was indicated by the presence of methylbenzylsuccinates with concentrations of 17-50 microg/L in three wells. Anaerobic toluene biodegradation was also indicated by benzylsuccinate concentrations of 10-49 microg/L in the three wells with the highest toluene concentrations (1500-5000 microg/L toluene). Since benzylsuccinate typically accounts for a very small fraction of the toluene present in groundwater (generally <1 mol%), the signature metabolite approach works best at higher toluene concentrations when it is not constrained by detection limits. In wells with lower toluene concentrations (410-640 microg/L), carbon and hydrogen isotopic values were enriched by up to approximately 2 per thousand for delta13C and approximately 70 per thousand for delta2H. This evidence of isotopic fractionation verifies the effects of biodegradation in these low concentration wells where metabolites may already be below detection limits. The combined use of signature metabolite and CSIA data is particularly valuable given the challenge of verifying biodegradation in subarctic environments where degradation rates are typically much slower than in temperate environments.
Effects of 12 biotic and abiotic factors on the freezing point of the sub-Antarctic springtail, Tullbergia antarctica, were investigated. Repeated cooling of individual springtails five times resulted in very similar freezing points suggesting that ice nucleation in this freeze-susceptible species is likely to be initiated by intrinsic factors rather than being a stochastic event. Mean supercooling point (SCP) was influenced by cooling protocol, showing a linear increase in mean SCP with cooling rates from 8 to 0.1 degrees Cmin(-1). However, the opposite effect (decreasing SCP) was seen with slower cooling. Slower rates may be ecologically realistic and allow time for appropriate physiological and biochemical changes. Feeding and food presence in the gut had no effect on SCP, and there was no correlation between the ice nucleating activity of bacteria isolated from the guts and the whole springtail SCP. Habitat altitude and diurnal light and temperature regimes also had no effect on SCP. There was no correlation between the cryoprotectant concentration of fresh animals and their SCP, but experimental desiccation resulted in increased osmolality and decreased SCP, although with considerable individual variation. The most significant influence on SCP was associated with ecdysis. As springtails cease feeding for a period either side of ecdysis, shedding the entire gut lining, moulting may be an efficient mechanism of clearing the gut of all ice nucleating material. This previously unrecognised relationship between ecdysis, cold tolerance and seasonal survival tactics may play an important role in over-winter survival of some arthropods.
Adaptation to both high and low temperatures requires proteins with special properties. While organisms living at or close to the boiling point of water need to have proteins with increased stability, other properties are required at temperatures close to the freezing point of water. Indeed, it has been shown that enzymes adapted to cold environments are less resistant to heat with a concomitant increased activity as compared to their warm-active counter-parts. Several recent studies have pointed in the direction that electrostatic interactions play a central role in temperature adaptation, and in this study we investigate the role such interactions have in adaptation of elastase from Atlantic salmon and pig. Molecular dynamics (MD) simulations have been used to generate structural ensembles at 283 and 310 K of the psychrophilic and mesophilic elastase, and a total of eight 12 ns simulations have been carried out. Even though the two homologues have a highly similar three-dimensional structure, the location and number of charged amino acids are very different. Based on the simulated structures we find that very few salt-bridges are stable throughout the simulations, and provide little stabilization/destabilization of the proteins as judged by continuum electrostatic calculations. However, the mesophilic elastase is characterized by a greater number of salt-bridges as well as a putative salt-bridge network close to the catalytic site, indicating a higher rigidity of the components involved in the catalytic cycle. In addition, subtle differences are also found in the electrostatic potentials in the vicinity of the catalytic residues, which may explain the increased catalytic efficiency of the cold-adapted elastase.
Uracil DNA glycosylase (UDG) is a DNA repair enzyme involved in the base excision repair (BER) pathway, removing misincorporated uracil from the DNA strand. The native and mutant forms of Atlantic cod and human UDG have previously been characterized in terms of kinetic and thermodynamic properties as well as the determination of several crystal structures. This data shows that the cold-adapted enzyme is more catalytically efficient but at the same time less resistant to heat compared to its warm-active counterpart. In this study, the structure-function relationship is further explored by means of comparative molecular dynamics (MD) simulations at three different temperatures (375, 400 and 425K) to gain a deeper insight into the structural features responsible for the reduced thermostability of the cold-active enzyme. The simulations show that there are distinct structural differences in the unfolding pathway between the two homologues, particularly evident in the N- and C-terminals. Distortion of the mesophilic enzyme is initiated simultaneously in the N- and C-terminal, while the C-terminal part plays a key role for the stability of the psychrophilic enzyme. The simulations also show that at certain temperatures the cold-adapted enzyme unfolds faster than the warm-active homologues in accordance with the lower thermal stability found experimentally.
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Selective brain cooling (SBC) can occur in hyperthermic humans despite the fact that humans have no carotid rete, a vascular structure that facilitates countercurrent heat exchange located at the base of the skull in some mammals. Emissary and angular veins, upper respiratory tract, tympanic cavity and cerebrospinal fluid are major components of SBC system in humans. The efficiency of SBC is increased by evaporation of sweat on the head and by ventilation through the nose, but it is surprising to find out that mammals do not display SBC during exercise hyperthermia. What is the explanation then for the SBC at high body temperatures? Our hypothesis is that selective brain cooling protects the brain from thermal damage in a long-standing manner by allowing adaptive mechanisms to change the craniofacial morphology appropriate for different environmental conditions. Since the brain can only be as big that can cool, it is not surprising to find a lower (below 1300 cm(3)) cranial volume in Australian Aborigines with respect to the one (over 1450 cm(3)) in Eskimos. In addition to lower brain volume, other craniofacial features such as thick everted lips, broader nasal cavity and bigger paranasal sinuses that provide more evaporating surfaces seem to be anatomical variations developed in time for an effective SBC in hot climates. It was reported previously that these biological adaptations result from the tissues of neural crest origin. Among the crest derivatives, leptomeninges (pia and arachnoid mater), skeletal and connective tissues of the face and much of the skull seem to be structures upon which environment operates to produce more convenient craniofacial morphology for an effective SBC. In conclusion, selective brain cooling seems to be a mechanism leading to adaptive craniofacial diversity observed in different geographical regions. Thus, SBC is necessary for long-term biological adaptation, not for protecting the brain from acute thermal damage.
Hundreds of isolates of viable bacteria and fungi have been recovered from ancient ice and permafrost. Evidence supports the hypothesis that viral pathogens also are preserved in ice repositories, such as glaciers, ice sheets, and lake ice. Proof may depend upon narrowing the search by applying specific criteria, which would target candidate viruses. Such criteria include viral pathogens likely to occur in great abundance, likely to be readily transported into ice, and then participate in ongoing disease cycles suggestive of their having been deposited in and subsequently released from ice. Caliciviruses, influenza A, and some enteroviruses appear to satisfy all three criteria. Environmental ice appears to be an important abiotic reservoir for pathogenic microbes. World health and eradication of specific pathogens could be affected by this huge reservoir.
AIMS: We studied the metabolic changes that took place in the crew of the Hesperides vessel in their 2001-2002 Antarctic journey, comparing two periods differing in diet and physical activity. METHODS AND RESULTS: Lipid profile, vitamin and hormone levels were analyzed in 17 subjects who completed the study in its two phases. In phase I the participants spent 47 days sailing with hard work and rough seas, and the diet was rich in fat and poor in fresh foods. In this phase, glucose decreased and HDL-cholesterol, apo-AI, and TSH increased. Plasma retinol and alpha-tocopherol levels remained stable, gamma-tocopherol, alpha-carotene and beta-carotene significantly decreased, and lycopene significantly increased. Phase II lasted 49 days including a 7-day long stop in port. This meant that a more varied diet was available and fresh foods were present in the hold. There was also less extreme physical activity. The metabolic pattern changed direction, glucose rose, HDL-cholesterol and apo-AI decreased and the levels of the vitamins that dropped in phase I started to increase. Lycopene significantly decreased. CONCLUSION: Contrary to popular beliefs about navigation at extreme latitudes, the metabolic changes described may be explained by the intense physical activity in a cold environment and a high-fat diet poor in fresh products.
Microbes have been discovered in permafrost sediments for nearly a century. However, microbiological analyses of alpine permafrost are very scarce. This study is a first attempt to describe the phylogenetic diversity of a culturable bacterial community isolated from alpine permafrost in the Tianshan Mountains in northwestern China. Aerobic 2.5-6.0x10(5) CFU/gdw (CFU per 1 gram of dry weight) on modified PYGV medium were recovered from alpine permafrost samples at 4 degrees C; among these, 91 bacterial isolates with different morphotypes were characterized by phenotypic properties, such as morphology, colony pigmentation, Gram staining, endospore formation and temperature range of growth. The isolates were further categorized based on amplified rDNA restriction analysis (ARDRA), and 51 representative isolates possessing distinct ARDRA patterns selected for subsequent 16S rDNA sequencing and phylogenetic analysis. The phylogenetic trees placed the 51 isolates in four major groups: the high-G+C Gram-positives, the low-G+C Gram-positives, Proteobacteria and the Cytophaga-Flavobacterium-Bacteroides (CFB) phylum. The most abundant and diverse isolates were members of Gram-positive bacteria, particularly the Arthrobacter as a dominant group in alpine permafrost culturable populations. Results of the Jukes-Cantor evolutionary distance matrix suggested that the vast majority of the isolates were different strains of known species, and three may represent new species within the genus Chryseobacterium of the CFB phylum. From this study, it is proposed that alpine permafrost sediments in the Tianshan Mountains provide a specific ecological niche for prolonging survival of diverse microbial lineages.
A combined household/industrial landfill in a humid and cold temperate climate was characterised with respect to its chemical composition. Cores taken at three randomly chosen sites on the landfill and at different depths at each site were analysed. Carbon, nitrogen and pH were measured by standard laboratory methods. The chemical elements analysed included metals and the non-metals B, P and S. pH ranged between 8.0 and 8.5. The total carbon content was in the interval 4.5-26.9% and the total nitrogen content in the interval 0.05-0.48%. The C/N ratio was high, indicating that there was not enough nitrogen available to ensure the stabilisation of carbon. The metal contents varied substantially. The water and carbon contents were related to each other as well as to the metal content, which increased with the content of water. Based on the results obtained regarding the chemical composition of the landfill, it is evident that the landfill consists of two layers. This indicates that the landfill body might have different levels of chemical development, due to water content, and different long-term leachability in the future.
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In this study we have measured exposure levels to quartz in different parts of the slate industry in Alta, Northern Norway. Full shift personal samples were collected from the breathing zones of outdoor and indoor workers in the slate quarries and a slate factory. The quartz content of respirable dust was between 7 and 41%. The slate factory had the lower quartz levels although 41% of total and 73% of respirable samples were above the Norwegian TLV for quartz. The average concentration of total quartz in the slate factory was 0.27 mg/m3 and the average concentration of respirable quartz was 0.12 mg/m3. Outdoor in the quarries the average levels of quartz were 0.58 and 0.13 mg/m3 for total and respirable quartz, respectively. From the beginning of the last decade most of the quarry-workers have built quarry halls to protect themselves against a cold winter climate. Inside in these quarry halls the average levels were 1.74 mg/m3 total quartz and 0.46 mg/m3 respirable quartz. Assessment of historical exposure showed that 32 of totally 45 quarry workers with available exposure history had a lifetime inhaled quartz dose of more than 10 g. There is reason to fear that silicosis will be an increasing problem among quarry workers if efforts to reduce quartz exposure are not put into effect.