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Membrane properties of an extreme thermophile. II. Membrane functions underlying leucine transport and their relation with thermotropic phase transitions.

Various membrane properties of Thermus thermophilus HB8 were studied in order to elucidate the mechanism and implication of cell adaptation to a high temperature environment. 1. The profile of temperature dependence of the amino acid uptake rates was similar for the membrane obtained from cells grown at both 61 degrees C and 77 degrees C. 2. The remaining uptake activity after heat treatment paralleled the membrane potential generating ability, as measured by auramine O fluorescence response to the respiration substrate, and was not dependent on the growth temperature. 3. The temperature dependence of the efflux of the accumulated amino acid reflects the physical phase of the membrane and is dependent on the growth temperature.

Biological Transport↗

Effect of working in hot environments on respiratory air temperatures.

The purpose of this study was to obtain an accurate measure of the temperature of respired air of subjects working on a treadmill at various ambient temperatures and ambient relative humidities (RH). The experiments were conducted in an environmental chamber at each of four different ambient temperatures (nominally 20, 30, 40 and 45 degrees C) and at two different ambient RH (20% and 100%) for a total of eight different conditions. Each experiment consisted of four tests at each ambient condition, these being: (A) standing quietly; (B) walking on a treadmill at 3 km.h-1, 0% grade; (C) walking on a treadmill at 3 km.h-1, 5% grade; and (D) walking on a treadmill at 5 km.h-1, 5% grade. It was found that under these conditions the maximum temperature of the expired air is independent of work rate and also ventilation rate but varies significantly with both the temperature and humidity of the inspired air. At low ambient RH the expired air temperature was [mean (SD)] 31.2 (1.3), 33.3 (0.7), 34.0 (1.7) and 35.6 (0.7) degrees C for ambient temperatures of 20, 30, 40 and 45 degrees C, respectively. At high ambient RH the expired air temperature was 32.0 (1.8), 35.3 (0.5), 37.6 (1.2) and 41.8 (0.8) degrees C at ambient temperatures of 20, 30, 40 and 45 degrees C, respectively. Thus the expired air temperature was higher at the higher ambient temperatures and ambient RH. While similar results have been reported before, the techniques used in this study should provide a more accurate measure of these effects than those previously reported.

Adult↗

Two thermosensors in Drosophila have different behavioral functions.

Insects inhabit extreme temperature environments and have evolved mechanisms to survive there. Small insects are especially susceptible to rapid changes in body temperature. Therefore, the rapid detection of environment and body temperature is important for their survival. Little, however, is known about the thermosensors that detect those temperatures. Using rapid thermosensitivity assays with temperature step gradients and a spatial learning paradigm (the heat-box) in which elevated temperature serves as the negative reinforcer, two thermosensors were identified and their behavioral functions assessed. A low-temperature thermosensor is located on the antenna, detects relatively low temperatures, and can detect spatial temperature gradients directly. Thus, the antennae can be used by Drosophila to quickly orient with respect to temperature cues. A high-temperature thermosensor of unknown location appears to have a roughly similar sensitivity to temperature differences as the low-temperature thermosensor (< or = 3 degrees C) and is both necessary and sufficient for memory formation in the heat-box spatial learning paradigm. Therefore, the high-temperature thermosensor is important for remembering spatial positions in which dangerously high temperatures were encountered.

Adaptation, Physiological↗

Cells transformed by temperature-sensitive mutants of avian sarcoma virus cause tumors in vivo at permissive and nonpermissive temperatures.

Chick embryo (CE) fibroblasts and normal rat kidney (NRK) cells transformed by temperature-sensitive (ts) mutants of avian sarcoma virus (NY68, LA23, LA24, LA25, LA29, LA31, GI201, GI202, GI251, GI253 induce tumors on the chorioallantoic membrane (CAM) of chick eggs at temperatures that correspond to the permissive and nonpermissive temperatures used to induce conditional expression of the "transformed" phenotype in these cells when cultured in vitro. Chick embryo cells infected with transformation-defective mutants of ASV (td101, td108) or RAV-50 were nontumorigenic under the same conditions, as were nontransformed CE and NRK cells. This indicates that the CAM is not an unusually susceptible substrate for cell growth and that the ability of tsASV-transformed cells to form tumors at nonpermissive temperatures reflects their true tumorigenicity. In contrast, a ts mutant chemically transformed rat liver cell line, ts-223, only formed tumors on the CAM under permissive conditions. The wild-type parent cells (W-8) of this mutant produced tumors at both permissive and nonpermissive temperatures. Direct implantation of microprobe thermometers into tumors caused by ts-ASV-transformed cells at nonpermissive temperatures confirmed that tumor formation occurred in a stable temperature environment and was not due to temperature fluctuations which might have created semi-permissive conditions for tumor growth. Cells isolated from tumors formed at nonpermissive temperatures and recultured in vitro displayed temperature-dependent hexose transport and colony formation in agar similar to the orginal parent cell inoculum. Similarly, virus recovered from tumors at nonpermissive temperatures retained the ts mutation.

Alpharetrovirus↗

Effect of insulated skin probes to increase skin-to-environmental temperature gradients of preterm infants cared for in convective incubators.

Thermal support systems for premature infants that are based on skin servocontrol depend on accurate measurement of skin surface temperature. We examined prospectively the effect of probe insulation to alter measured skin temperature in 10 preterm infants. The use of insulated probes resulted in significant alteration in incubator servocontrol, with lower incubator air temperatures and higher skin-to-environment temperature gradients.

Equipment Design↗

Thermal noise reduction of mechanical oscillators by actively controlled external dissipative forces

We show that the thermal fluctuations of very soft mechanical oscillators, such as the cantilever in an atomic force microscope (AFM), can be reduced without changing the stiffness of the spring or having to lower the environment temperature. We derive a theoretical relationship between the thermal fluctuations of an oscillator and an actively controlled external dissipative force. This relationship is verified by experiments with an AFM cantilever where the external active force is coupled through a magnetic field. With simple instrumentation, we have reduced the thermal noise amplitude of the cantilever by a factor of 3.4, achieving an apparent temperature of 25 K with the environment at 295 K. This active noise reduction approach can significantly improve the accuracy of static position or static force measurements in a number of practical applications.

Journal Article↗

Contraction rhythm in the plasmodium of Physarum polycephalum: dependence of the period on the amplitude, temperature and chemical environment.

The plasmodium of Physarum polycephalum exhibits a characteristic protoplasmic shuttle streaming and generates periodical tension. We determined the factors which govern the period of the contraction rhythm by measuring isometric tension of the plasmodial strand and the motive force of the protoplasmic streaming under a variety of conditions by changing the size of plasmodia, chemical composition of environment and surrounding temperatures. The results are: (1) The period of contraction rhythm, tau, increased linearly with the amplitude of oscillating tension, F, and was expressed by the following empirical equation when F was lower than a certain critical value, Fc, i.e., tau = aF+tau O. Above Fc, tau stayed at a constant level of tau S. There, a tau O and tau S are numerical constants which are independent of F. A similar relationship is valid for the amplitude of the motive force of the protoplasmic streaming, delta P. (2) Values of tau O were 1.0 min in air and 1.6 min in an aqueous medium and they were independent of temperature. (3) The Arrhenius plots of the parameter "a" exhibited different straight lines in air and in aqueous medium, from which the values of Q10 were determined to be 4.0 and 10, respectively. (4) Lowering of temperature decreased Fc, and eventually diminished it at Tc (= 15 degrees C). (5) The presence of glucose, CaCl2, MgCl2 and NaCl gave the identical tau-F relationship. Applications od D2O and ethanol slowed down the contraction rhythm, while that of KCl accelerated the rhythm. However, all these chemicals did not affect the tau O value. All results described above indicate that the contraction rhythm in the plasmodium is influenced by three components: a limit cycle which is independent of tension generation, a component which is strongly linked to the amplitude of the tension, F, and the part which is independent of the value of F. External factors appear to influence separately these three components of the contraction rhythm.

Cytoplasmic Streaming↗

Assessment of the biodegradation potential of psychrotrophic microorganisms.

Bioremediation of polluted temperate and cold temperature environments may require the activity of psychrotrophic bacteria, because their low temperature growth range parallels the ambient temperatures encountered in these environments. In the present study, 135 psychrotrophic microorganisms isolated from a variety of ecosystems in Canada were examined for their ability to mineralize 14C-labelled toluene, naphthalene, dodecane, hexadecane, 2-chlorobiphenyl, and pentachlorophenol. A number of the psychrotrophic strains mineralized toluene, naphthalene, dodecane, and hexadecane. None of the psychrotrophs were capable of mineralizing 2-chlorobiphenyl or pentachlorophenol. Those strains demonstrating mineralization activity were subsequently screened by the polymerase chain reaction (PCR) and Southern hybridization of PCR products for the presence of catabolic genes (alkB, ndoB, todCl, and xylE) involved in known bacterial biodegradative pathways for these compounds. Some of the psychrotrophs able to mineralize toluene and naphthalene possessed catabolic genes that hybridized to xylE or todCl, and ndoB, respectively. The alkB PCR fragments obtained from the strains that mineralized dodecane and hexadecane did not hybridize to an alkB gene probe derived from Pseudomonas oleovorans. Psychrotrophic strain Q15, identified as a Rhodococcus sp., also mineralized the C28 n-paraffin octacosane. A gene probe constructed from the "alkB" PCR fragment from strain Q15 did hybridize with the alkB PCR fragments from most of the psychrotrophic alkane biodegraders, indicating that the alkB primers may be amplifying another gene(s), perhaps with low homology to P. oleovorans alkB, which may be involved in the biodegradation of both short chain (dodecane) and longer chain alkanes (hexadecane, octacosane). All of the psychrotrophic biodegradative isolates examined were capable of mineralization activity at both 23 and 5 degrees C, indicating their potential for low temperature bioremediation of petroleum hydrocarbon contaminated sites.

Arthrobacter↗

[Experimental analysis of mechanism concerning the environmental temperature effect on acquisition of capability in toad oocyte to resume meiotic division].

Full-grown oocytes derived from Bufo bufo gargarizans rearing in high temperature environment (28-30 degrees C), called high temperature oocytes, never underwent germinal vesicle breakdown (GVBD) after progesterone stimulation, no MPF was detected in their ooplasm, but some events which manifested normally at the beginning of progesterone induced maturation process were revealed in these oocytes. It is worth notice an another kind of maturation promoting substance(s) appeared in the ooplasm of high temperature oocyte after the hormone treatment, which was capable of triggering the resumption of meiotic division of the full-grown oocytes derived from hibernating toad (called low temperature oocytes). It is a hibernation factor-dependent maturation promoting substance (HF-MPS), which appeared after decrease of the oocyte endogenous cAMP level. Its appearance depended upon the oocyte protein synthesis, and its activity to inducing GVBD of low temperature oocytes did not inhibited by puromycin. HF-MPS differs from MPF in maturation promoting activity, as low temperature (10 degrees C) delayed obviously HF-MPS' activity but did not influence the rate of GVBD induced by MPF. Further more, probably due to the lack of "hibernation factor(s)", no expression of p34cdc2 gene was detected in high temperature oocytes (unpublished data), neither HF-MPS nor MPF could amplify autocatalytically in the oocytes. So the low temperature (below 15 degrees C) was found to be indispensable for the toad oocyte maturation. If one day we can prove HF-MPS appeared also in the course of oocyte maturation induced by progesterone, the relationship between HF-MPS and MPF may be: [formula: see text] All these discoveries indicated above make a reasonable explanation of the geographical distribution of the toad which was restricted in the region north to the 23 degrees north latitude and east to the 100 degrees east longitude in China.

Animals↗

Monitoring of fuel consumption and aromatics formation in a kerosene spray flame as characterized by fluorescence spectroscopy.

The large presence of aromatic compounds in distillate fossil fuels should allow, in line of principle, to follow the fuel consumption and/or the presence of unburned fuel in a high temperature environment like a burner or the exhaust of combustion systems by exploiting the high fluorescence emission of aromatic fuel components. To this aim an UV-excited fluorescence source has to be used since the aromatic fuel components are strongly fluorescing in the UV region of the emission spectrum. In this work UV-excited laser induced fluorescence (LIF) diagnostics was applied to spray flames of kerosene in order to follow the fuel consumption and the formation of aromatic species. A strong UV signal was detected in the spray region of the flame that presented a shape similar to that found in the LIF spectra preliminary measured on the cold spray and in the room-temperature fluorescence of fuel solutions. The decrease of UV signal along the spray flame region was associated to the consumption of the fuel, but more difficult seems to be the attribution of a broad visible emission, that is present downstream of the flame. The visible emission feature could be assigned to flame-formed PAH species contained in the high molecular weight species, hypothesizing that their fluorescence spectra are shifted toward the visible for effect of the high temperature flame environment.

Incineration↗

Thermoregulation in Yersinia enterocolitica is coincident with changes in DNA supercoiling.

Yersinia enterocolitica is a facultative intracellular parasite, displaying the ability to grow saprophytically or invade and persist intracellularly in the mammalian reticuloendothelial system. The transition between such diverse environments requires the co-ordinated regulation of specific sets of genes on both the chromosome and virulence plasmid. Temperature has a profound pleiotropic effect on gene expression and phenotypically promotes alterations in cell morphology, outer-membrane protein synthesis, urease production, lipopolysaccharide synthesis, motility, and synthesis of genes involved in invasion of eukaryotic host cells. By examining thermoregulated flagella biosynthesis, we have determined that motility is repressed at 25 degrees C (permissive temperature) with subinhibitory concentrations of novobiocin. These conditions also induce virulence gene expression suggesting novobiocin addition simulates, at least partially, a high-temperature environment. Furthermore, temperature-shift experiments, using Y. enterocolitica containing pACYC184 as a reporter plasmid, indicate that thermo-induced alterations of DNA supercoiling coincide with temperature-induced phenotypic changes. A class of putative DNA gyrase mutant (novobiocin resistant) likewise demonstrates the 37 degrees C phenotype when cultured at 25 degrees C; it is non-motile, urease negative, calcium growth dependent, and positive for Yop expression. These results support a model implicating DNA topology as a contributing factor of Y. enterocolitica thermoregulation.

Bacterial Outer Membrane Proteins↗

Testicular migration, spermatogenesis, temperature regulation and environment of the sheath-tail bat, Taphozous georgianus.

The testes of the common sheath-tail bat of tropical Australia undergo a seasonal migration between the abdomen and the scrotal pouches, while each cauda epididymidis is permanently maintained in the scrotal pouch. Straps of smooth muscle attach to both the cranial and caudal poles of the testes, and these extend cranially to the diaphragm and caudally to the cauda epididymidis. The testicular arteries are not coiled. Among the environmental factors investigated, maximum temperature correlated most significantly with testicular descent, and the number of spermatogonia per bat also correlated most significantly with maximum temperature. Body temperature of a captive bat ranged from 25 to 38 degrees C and this was closely related to body weight and ambient temperature. It seems likely that the scrotal pouch provides a temperature slightly below that of the body and so facilitates sperm storage in the permanently scrotal cauda epididymidis. Migration of the testes probably serves to ameliorate the seasonal temperature fluctuations to which they are exposed while the relatively high correlation between maximum environment temperature and spermatogonial numbers suggests that temperature may be a proximate influence on reproduction in the sheath-tail bat.

Animals↗

Choroidal blood flow as a heat dissipating mechanism in the macula.

We studied the possible role of the choroidal circulation in stabilizing the temperature environment of the retina and the retinal pigment epithelium. Temperature measurements were taken from the retina and choroid of the monkey eye while the ambient illumination and choroidal blood flow were varied. The results showed that the choroidal circulation plays a significant role in dissipating heat generated by the focusing of light at the macula and suggest that this function may be a primary reason for the relatively high values of choroidal blood flow.

Animals↗

High-temperature Raman spectroscopy of solid oxide fuel cell materials and processes.

Chemical and material processes occurring in high temperature environments are difficult to quantify due to a lack of experimental methods that can probe directly the species present. In this letter, Raman spectroscopy is shown to be capable of identifying in-situ and noninvasively changes in material properties as well as the formation and disappearance of molecular species on surfaces at temperatures of 715 degrees C. The material, yttria-stabilized zirconia or YSZ, and the molecular species, Ni/NiO and nanocrystalline graphite, factor prominently in the chemistry of solid oxide fuel cells (SOFCs). Experiments demonstrate the ability of Raman spectroscopy to follow reversible oxidation/reduction kinetics of Ni/NiO as well as the rate of carbon disappearance when graphite, formed in-situ, is exposed to a weakly oxidizing atmosphere. In addition, the Raman active phonon mode of YSZ shows a temperature dependent shift that correlates closely with the expansion of the lattice parameter, thus providing a convenient internal diagnostic for identifying thermal gradients in high temperature systems. These findings provide direct insight into processes likely to occur in operational SOFCs and motivate the use of in-situ Raman spectroscopy to follow chemical processes in these high-temperature, electrochemically active environments.

Letter↗