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Comparative sequence and structure analysis reveal features of cold adaptation of an enzyme in the thermolysin family.

Knowledge about the structural features underlying cold adaptation is important for designing enzymes of different industrial relevance. Vibriolysin from Antarctic bacterium strain 643 (VAB) is at present the only enzyme of the thermolysin family from an organism that thrive in extremely cold climate. In this study comparative sequence-structure analysis and molecular dynamics (MD) simulations were used to reveal the molecular features of cold adaptation of VAB. Amino acid sequence analysis of 44 thermolysin enzymes showed that VAB compared to the other enzymes has: (1) fewer arginines, (2) a lower Arg/(Lys + Arg) ratio, (3) a lower fraction of large aliphatic side chains, expressed by the (Ile + Leu)/(Ile + Leu + Val) ratio, (4) more methionines, (5) more serines, and (6) more of the thermolabile amino acid asparagine. A model of the catalytic domain of VAB was constructed based on homology with pseudolysin. MD simulations for 3 ns of VAB, pseudolysin, and thermolysin supported the assumption that cold-adapted enzymes have a more flexible three-dimensional (3D) structure than their thermophilic and mesophilic counterparts, especially in some loop regions. The structural analysis indicated that VAB has fewer intramolecular cation-pi electron interactions and fewer hydrogen bonds than its mesophilic (pseudolysin) and thermophilic (thermolysin) counterparts. Lysine is the dominating cationic amino acids involved in salt bridges in VAB, while arginine is dominating in thermolysin and pseudolysin. VAB has a greater volume of inaccessible cavities than pseudolysin and thermolysin. The electrostatic potentials on the surface of the catalytic domain were also more negative for VAB than for thermolysin and pseudolysin. Thus, the MD simulations, the structural patterns, and the amino acid composition of VAB relative to other enzymes of the thermolysin family suggest that VAB possesses the biophysical properties generally following adaptation to cold climate.

Acclimatization↗

Tracheal tubes in cold stress.

During rescues in cold climates, plastic tracheal tubes have fractured because of cold stress or mishandling. Susceptibility of rubber and plastic tracheal tubes to cold stress was tested in a climate chamber. Plastic tubes became stiffer when chilled, had greater resistance to reshaping and were more sensitive to shock and manipulation. They also need protection against cold and shock during transport. Rubber tubes were more tolerant of cold stress. Very cold rubber and plastic tubes may induce tissue damage. Rewarming to a nondangerous temperature was shown to be a quick procedure. It is suggested that the rescuer's own body heat be used.

Cold Climate↗

The adaptive significance of reptilian viviparity in the tropics: testing the maternal manipulation hypothesis.

Phylogenetic transitions from oviparity to viviparity in reptiles generally have occurred in cold climates, apparently driven by selective advantages accruing from maternal regulation of incubation temperature. But why, then, are viviparous reptiles so successful in tropical climates? Viviparity might enhance fitness in the tropics via the same pathway as in the temperate zone, if pregnant female reptiles in the tropics maintain more stable temperatures than are available in nests (Shine's maternal manipulation hypothesis). Alternatively, viviparity might succeed in the tropics for entirely different reasons than apply in the temperate zone. Our data support the maternal manipulation hypothesis. In a laboratory thermal gradient, pregnant death adders (Acanthophis praelongus) from tropical Australia maintained less variable body temperatures (but similar mean temperatures) than did nonpregnant females. Females kept at a diel range of 25-31 degrees C (as selected by pregnant females) gave birth earlier and produced larger offspring (greater body length and head size) than did females kept at 23-33 degrees C (as selected by nonpregnant snakes). Larger body size enhanced offspring recapture rates (presumably reflecting survival rates) in the field. Thus, even in the tropics, reproducing female reptiles manipulate the thermal regimes experienced by their developing embryos in ways that enhance the fitness of their offspring. This similarity across climatic zones suggests that a single general hypothesis--maternal manipulation of thermal conditions for embryogenesis--may explain the selective advantage of viviparity in tropical as well as cold-climate reptiles.

Adaptation, Physiological↗

Precise dating of Dansgaard-Oeschger climate oscillations in western Europe from stalagmite data.

The signature of Dansgaard-Oeschger events--millennial-scale abrupt climate oscillations during the last glacial period--is well established in ice cores and marine records. But the effects of such events in continental settings are not as clear, and their absolute chronology is uncertain beyond the limit of (14)C dating and annual layer counting for marine records and ice cores, respectively. Here we present carbon and oxygen isotope records from a stalagmite collected in southwest France which have been precisely dated using 234U/230Th ratios. We find rapid climate oscillations coincident with the established Dansgaard-Oeschger events between 83,000 and 32,000 years ago in both isotope records. The oxygen isotope signature is similar to a record from Soreq cave, Israel, and deep-sea records, indicating the large spatial scale of the climate oscillations. The signal in the carbon isotopes gives evidence of drastic and rapid vegetation changes in western Europe, an important site in human cultural evolution. We also find evidence for a long phase of extremely cold climate in southwest France between 61.2 +/- 0.6 and 67.4 +/- 0.9 kyr ago.

Animals↗

Making human nasal cilia beat in the cold: a real time assay for cell signalling.

Human nasal epithelium must adapt to cold climates, and yet, in vitro, human ciliary beat frequency (CBF) is zero at 4 degrees C. Similarly, hibernating mammals do not die of pneumonia despite a core body temperature as low as 6 degrees C, implying that cilia continue to beat. Here, we show that protein kinase C (PKC) and Ca(2+)/calmodulin-dependent kinase II (CaMKII) regulate the profile of human nasal CBF in response to rising temperature from 4 degrees C. Onset of ciliary beat was at 10 degrees C in Medium 199, 7 degrees C in the presence of the PKC activator phorbol 12-myristate 13-acetate (PMA), the calcium ionophore ionomycin, or the CAMKII blocker myristoylated autocamtide-2 related inhibitory peptide (MACI), and 6 degrees C for the myristoylated peptide PKC inhibitor EGF-R Fragment 651-658 (MyrPKCI). During cell warming to 32 degrees C, the thermal profile was sigmoid in all solutions except those containing MACI+PMA. Surprisingly, cilia continued to beat despite 4 degrees C and were significantly more responsive to rising temperature with either MACI+PMA, or MACI+MyrPKCI. Our data suggest that CaMKII and PKC regulate the thermal slope and profile of CBF in vitro, and that when these protein kinases are manipulated, cilia can continue to beat despite hypothermia. These findings may relate to adaptive responses to cold climates.

Adaptation, Physiological↗

Internal nasal floor configuration in Homo with special reference to the evolution of Neandertal facial form.

The presence of a steeply sloping or depressed nasal floor within the nasal cavity of Neandertals is frequently mentioned as a likely specialization or autapomorphy. The depressed nasal floor has also been seen as contributing to a relatively more capacious nasal cavity in Neandertals, which is tied to cold-climate respiratory adaptation and energetics. These observations have been limited largely to a relatively few intact crania, and the character states associated with this trait have not been as precisely codified or analyzed as those published for Plio-Pleistocene hominins (McCollum et al., 1993, J. Hum. Evol. 24, 87; McCollum, 2000, Am. J. Phys. Anthrop. 112, 275). This study examines the internal nasal floor topography in complete crania and isolated maxillae in European, west Asian, and African fossil Homo (n=158) including 25 Neandertals, and a wide range of recent humans from Europe, the Near East, and Africa (n=522). The configuration of the internal nasal floor relative to the nasal cavity entrance is codified as: 1) level, forming a smooth continuous plane; 2) sloped or mildly stepped; or 3) bilevel with a pronounced vertical depression. The frequency of these nasal floor configurations, and their relationship to both nasal margin cresting patterning and a comprehensive set of nasofacial metrics is examined. Neandertals show a high frequency of the bilevel (depressed) configuration in both adults and subadults (80%), but this configuration is also present in lower frequencies in Middle Pleistocene African, Late Pleistocene non-Neandertal (Skhul, Qafzeh), and European Later Upper Paleolithic samples (15%-50%). The bilevel configuration is also present in lower frequencies (ca. 10%) in all recent human samples, but attains nearly 20% in some sub-Saharan African samples. Across extinct and extant Homo (excluding Neandertals), internal nasal floor configuration is not associated with piriform aperture nasal margin patterning, but the two are strongly linked in Neandertals. Variation in internal nasal floor configuration in recent humans is primarily associated with internal nasal fossa breadth and nasal bridge elevation, whereas in fossil hominins, it is associated primarily with variation in facial height. Cold-climate and activity-related thermal adaptation as an explanation for the high frequency of pronounced nasal floor depression in Neandertals is inconsistent with all available data. Alternatively, variation in internal nasal floor configuration is more likely related to stochastically derived populational differences in fetal nasofacial growth patterns that do not sharply differentiate genus Homo taxa (i.e., cladistically), but do phenetically differentiate groups, in particular the Neandertals, especially when considered in combination with other nasofacial features.

Adaptation, Physiological↗

Effects of temperature on vibration-induced damage in nerves and arteries.

Vasospastic episodes in hand-arm vibration syndrome are more prevalent among power-tool workers in cold climates. To test whether cold enhances vibration-induced damage in arteries and nerves, tails of Sprague-Dawley rats were vibrated at room temperature (RT) or with tail cooling (<15 degrees C). Cold vibration resulted in a colder tail than either treatment alone. Vibration at both temperatures reduced arterial lumen size. RT vibration generated more vacuoles in arteries than cold vibration. Vibration and cold induced nitration of tyrosine residues in arteries, suggesting free-radical production. Vibration and cold generated similar percentages of myelinated axons with disrupted myelin. Cold with and without vibration caused intraneural edema and dilation of arterioles and venules with blood stasis, whereas vibration alone did not. The similarities, differences, and interactive effects of cold and vibration on nerve and artery damage indicate that temperature is involved mechanistically in the pathophysiology of hand-arm vibration syndrome.

Animals↗

The ScanDeNi process could turn an existing under-performing activated sludge plant into an asset.

With tightening up of effluent discharge standards from wastewater treatment facilities, many plants are facing costly augmentations and in many cases completely new plants will have to be constructed. The ScanDeNi process was developed in Sweden for increased nitrogen removal at the Västerås Sewage Treatment Plant (STP), 125,000 p.e. near Stockholm, and can be described as a modified contact stabilisation process with pre-denitrification and a selector stage for nitrification. The STP was upgraded at a cost of some 25 Mill. SEK (2.5 Mill. USD). It has been successfully in operation since 1998, exceeding all expectations. The process is showing the following major advantages. 25-35% less volume for the same Sludge Retention Time (SRT) and secondary sedimentation sludge load, compared to conventional pre-denitrification; or a 25-35% higher load can be applied within the same volume with the same removal efficiencies. The selector mechanism appears to be not limited to the nitrifying bacteria alone. Other microorganisms appear to be responsible for the reduction of surface active matter from the return activated sludge (RAS), as well as in the reject stream from sludge dewatering, resulting in an increase in alpha-values of approximately 50%. Due to the high alpha-values less aeration is required, resulting in significant operating cost savings. 'Automatic' creation of anaerobic conditions, enabling biological phosphorus removal. Whilst rarely a concern in warmer climates, BNR plants in cold climates in winter often lose their capacity to nitrify. The Västerås STP has consistently maintained excellent effluent quality even with effluent temperatures as low as 7 degrees C, and at an SRT of some 7-9 days, proving the effectiveness of the nitrifier selector. The ScanDeNi process could offer excellent effluent discharge standards (T-N < 10 mg/L, T-P < 0.5 mg/L) in smaller tank volumes and at a significantly lower operating cost, compared to conventional pre-denitrification systems.

Automation↗

Mummified seal carcasses in the McMurdo Sound Region, Antarctica.

Information was collected on 90 mummified carcasses of the "crabeater" seal in the ice-free areas of the McMurdo Sound region, Antarctica. The carcasses range from relatively well-preserved bodies to merely old, twisted, winddissected fragments of tissue. They are hard and dry and lie on the surface of the ground, mostly in valley bottoms. The arid, cold climate is ideal for retarding organic decay. One carcass was dated by radiocarbon analysis and found to be between 1600 and 2600 years old.

Antarctic Regions↗

Geographic variation in the prevalence of Raynaud's phenomenon: a 5 region comparison.

OBJECTIVE: To determine the population based prevalence of Raynaud's phenomenon (RP) in 5 geographic regions: one in South Carolina, USA, and 4 in France; to explore the relationship of RP to the climate; to investigate possible risk factors; and to describe the characteristics of RP+ subjects in the general population. METHODS: The study consisted of 2 phases: a telephone survey of a randomly drawn sample of households, with 10,149 completed interviews; these were followed by a face to face interview and clinical evaluation (n = 1,534), including diagnosis of RP. The same methodology was used in all 5 regions: for recruitment of subjects, criteria for RP, method of RP diagnosis, and for gathering additional information. RESULTS: The prevalence of RP was found to be related to the climate. The relationship between RP and climate was complicated, however, by the fact that many subjects had moved between climate zones. The relationship of RP to a cold climate became more evident after taking the migration patterns into account: the majority of RP+ subjects in the 2 coldest regions had lived all their lives in the same or a similar climate zone; the majority of RP+ subjects in the 2 warmest regions had previously lived in a colder climate. Other factors associated with RP were family history of RP, cardiovascular diseases, older age, a low body mass index, use of vibrating tools, and outings of a day or more. The classical triphasic RP was rarely encountered in the general population and the most frequently observed signs and symptoms during an RP attack were blanching accompanied by numbness. CONCLUSION: In addition to being a triggering factor for RP attacks, cold also appears to be an etiologic factor in the pathogenesis of RP. A subclinical cold injury, more likely to occur in colder climates, may be responsible for the "local fault" that has been implicated in the pathogenesis of RP and, in association with other risk factors, may predispose subjects to develop clinical RP.

Activities of Daily Living↗

High coronary mortality in cold regions of Sweden.

The hypothesis that cold climate is associated with high coronary mortality in Sweden is tested. Cold exposure was calculated in each of the 284 municipalities of Sweden. There was a significant association between cold exposure and coronary mortality in both sexes in all age groups. The strongest association was found in men aged 40-64 years (coefficient of determination k = 0.39). The decile of men aged 40-64 years who lived in the coldest municipalities had a 40% excess mortality. A significant association was also found between cold exposure and mortality from cerebrovascular diseases. We conclude that there is a strong regional association between cold exposure and high coronary mortality.

Adult↗

Resultant clothing insulation during exercise in the cold.

Predictions of required clothing during work in the cold is often based on data obtained from thermal manikin measurements. These clothing basic insulation values are altered during exercise. In a series of experiments at our laboratory, resultant clothing insulation has been studied. In a cold climatic chamber subjects walked on a treadmill at various exercise intensities dressed in cold protective clothing at different levels of insulation. Thermal insulation of the clothing ensembles was measured with a thermal manikin under standardized conditions. Compared to the standard manikin insulation values, the clothing insulation values measured (resultant insulation) at the human experiments were always lower. Reductions of insulation ranged from 12 to 54% depending on such factors as work intensity, sweating amount, clothing type etc. Predictions of cold protection of clothing at actual use must take into account factors influencing clothing insulation to avoid over-estimation of the cold protective properties of clothing.

Clothing↗

Reptile freeze tolerance: metabolism and gene expression.

Terrestrially hibernating reptiles that live in seasonally cold climates need effective strategies of cold hardiness to survive the winter. Use of thermally buffered hibernacula is very important but when exposure to temperatures below 0 degrees C cannot be avoided, either freeze avoidance (supercooling) or freeze tolerance strategies can be employed, sometimes by the same species depending on environmental conditions. Several reptile species display ecologically relevant freeze tolerance, surviving for extended times with 50% or more of their total body water frozen. The use of colligative cryoprotectants by reptiles is poorly developed but metabolic and enzymatic adaptations providing anoxia tolerance and antioxidant defense are important aids to freezing survival. New studies using DNA array screening are examining the role of freeze-responsive gene expression. Three categories of freeze responsive genes have been identified from recent screenings of liver and heart from freeze-exposed (5h post-nucleation at -2.5 degrees C) hatchling painted turtles, Chrysemys picta marginata. These genes encode (a) proteins involved in iron binding, (b) enzymes of antioxidant defense, and (c) serine protease inhibitors. The same genes were up-regulated by anoxia exposure (4 h of N2 gas exposure at 5 degrees C) of the hatchlings which suggests that these defenses for freeze tolerance are aimed at counteracting the injurious effects of the ischemia imposed by plasma freezing.

Animals↗

Comparison of the indoor air quality in mould damaged and reference buildings in a subarctic climate.

The purpose of this study was to search for objective parameters most relevant to indicate microbial problems of buildings in cold climate. Various indoor characteristics were compared in nine buildings with known history of moisture problems and visible mould (index) and in nine matched reference buildings. The concentrations of airborne viable fungal had a clear difference between the two groups of buildings. In this study, airborne concentrations of viable bacteria, formaldehyde, total volatile organic compounds (TVOC) and the occurrence of house dust mites in these index buildings were compared with the levels of the pollutants in matched reference buildings. Fungal growth and flora on moist building materials were also studied. The concentrations of TVOC were slightly higher in the index buildings than in the reference buildings. However, the differences in the concentrations or appearance of any of the studied pollutants were not significant. These parameters do not seem to be relevant indicators of microbial growth or surrogates of microbial exposure. Thus, fungal concentration and composition of fungal genera in the air still seems to be the best indicator for moisture problems among the studied pollutants. In the moist building materials, some fungal genera, such as Ulocladium and Chaetophoma were detected that were not found in indoor air showing that building material samples give additional information on the microflora of the building.

Actinomycetales↗

Field investigation of advanced filtration for phosphorus removal from constructed treatment wetland effluents.

Three sorptive media, blast furnace slag, cement clinker, and gravel were investigated for their capacity to remove phosphorus in a subsurface flow constructed treatment wetland post-treatment filter. Three filters were designed: two containing a mixture of either slag and gravel or clinker and gravel, and one with gravel only as the control filter. They were installed as a demonstration polishing treatment step to a constructed treatment wetland treating residential wastewater collected from 137 mobile home units at the Sunny Creek Estates Mobile Home Park. The filters were commissioned during the summer of the field season, with the data gathering taking place over a period of one year. The slag filter consistently had the lowest outflow dissolved phosphorus concentrations (0.27 +/- 0.08 mg l(-1), n=21), whereas the clinker filter had the highest outflow dissolved phosphorus concentrations (0.72 +/- 0.20 mg l(-1), n=23). The clinker filter performed well below expectations based on previous laboratory investigations, possibly due to lower pH conditions encountered in the field study. All of the filters maintained relatively constant outflow concentrations of phosphorus below 1 mg l(-1), despite varying input conditions and environmental factors such as temperature and phosphorus loading, and the occurrence of net export of phosphorus from the wetland. Net export of phosphorus from the filters occurred in the winter months, which was attributed to the decrease in input phosphorus concentrations below the maintained outflow concentrations (leading to phosphorus desorption), rather than cold temperatures or failure of the filter. Although the exact mechanisms of phosphorus removal are still under investigation, the results from this field study allowed for recommendations to be made for improving phosphorus removal at the Sunny Creek Estates constructed treatment wetland, in addition to providing valuable information for new and existing constructed treatment wetlands in cold climate areas.

Adsorption↗