Alterations in water intake and core temperature in baboons during hypothalamic thermal stimulation.
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In nineteen alpha-chloralose-anaesthetized 3- to 8-day-old lambs, the effect of cooling focally in the region of the locus coeruleus on respiratory output, as measured from phrenic nerve activity, was examined during normoxia and isocapnic hypoxia, and during infusion of adenosine or saline. Eleven lambs showed a 'biphasic' respiratory response to hypoxia, with an initial increase in phrenic nerve activity (phase 1) being followed by a decline to prehypoxic levels (phase 2). Cooling to 26-34 degrees C unilaterally in the locus coeruleus reversed the fall in respiratory output. When the probe temperature was returned to body temperature (39 degrees C) respiratory output fell again to phase 2 levels. Cooling had no effect during normoxia. In these lambs, adenosine infusion (0.3 mg kg-1 min-1 intra-arterially) during normoxia also decreased respiratory output. During adenosine infusion, cooling again increased respiratory output. Saline infusion did not affect respiratory output during normoxia with or without cooling. The other lambs did not show a biphasic respiratory response, i.e. increased respiratory output was maintained during isocapnic hypoxia. In these lambs adenosine had no effect on respiratory output. Cooling/rewarming the probe did not affect respiratory output during normoxia or hypoxia, or during adenosine or saline infusion. These results show that focal cooling in the dorsal pons at the locus coeruleus removes the inhibition of respiratory output seen in the neonatal biphasic respiratory response to hypoxia, but does not affect breathing in normoxia. This suggests that an important component of the biphasic respiratory response is mediated by the activation of neurones inhibitory to respiratory output, and involves either axons of passage or cell bodies lying in the locus coeruleus. Furthermore, the inhibitory effects of adenosine on breathing in the neonate appear to be mediated via this site.
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1. The rate of blood flow through the tail of the pig has been measured by means of venous occlusion plethysmography using a mercury in rubber strain gauge. Conscious animals were used in all experiments because previous work had demonstrated that anaesthetics interfere with the animal's ability to vasoconstrict.2. Graded changes in the temperature of the hypothalamus were imposed by means of an implanted thermode. It was found that the change in blood flow depended on the extent of the change in hypothalamic temperature and on the ambient temperature. Below 20 degrees C ambient temperature, heating the hypothalamus did not cause vasodilatation and at 30 degrees C ambient temperature, cooling the hypothalamus caused only slight vasoconstriction, but at 25 degrees C ambient temperature, changes in hypothalamic temperature caused changes in blood flow from full vasoconstriction to full vasodilatation.3. The skin temperature on the trunk was changed by means of water circulated through tubes sewn into a coat worn by the pig. Blood flow in the tail, which was outside the coat, depended on the skin temperature of the trunk, the ambient temperature, and the temperature of the hypothalamus, all of which were varied separately.4. A thermode was implanted in the epidural space in the cervical region of the spinal cord. The change in blood flow in the tail which accompanied a change in thermode temperature was found to depend on the temperature of the thermode and the ambient temperature. Cooling the spine while the hypothalamus was being heated to 43 degrees C resulted in a decrease in blood flow, but when the spine was heated while the hypothalamus was being cooled the increase in blood flow was only slight.5. Local stimulus to the tail in the form of infra-red heat, or increased air movement was followed by changes in blood flow even when deep body temperature remained stable.
The effects of a thermally-significant blood vessel, simulated by an embedded acrylic tube, 4.8 mm outer diameter on the freezing field caused by a surface cryoprobe were studied experimentally in a tissue phantom. The flat, 15 mm diameter, circular cryoprobe was operated at a constant cooling rate of -8 degrees C/min by liquid nitrogen down to -60 degrees C. Water flow rates of 30 and 100 ml/min, at a constant temperature of 32.5 degrees C, were maintained in the embedded tube. The latter flow rate is typical to the lower range of blood flows in large arteries in the human body. The phase changing medium (PCM) used was a 30/70% by volume mashed potatoes flakes-water solution. Temperature measurements inside the PCM were performed in one plane perpendicular to the embedded tube, relative to which the cryoprobe was placed at 5 locations in separate experiments. This novel experimental method reduced the perturbation caused by the thermocouple junctions while facilitating rather detailed measurements of the temperature fields developing in the PCM. Results show the development of two hump-like formations on either side of the embedded tube. Freezing was retarded in the region away from the surface cryoprobe and under the tube. This accentuated the dominance of the axial effects, due to the embedded tube, over the radial ones due to the cryoprobe. Results of this study should be considered in designing protocols of cryosurgical procedures performed in the vicinity of thermally-significant blood vessels.
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At an ambient temperature of 25 degrees C, selective cooling of the area preoptica medialis to 24 degrees +/- 1 degrees C produced a significant decrease in food intake together with hyperthermia. Heating the same area to 43 degrees +/- 1 degrees C resulted in the opposite effects. At an ambient temperature of 35 degrees C, heating the area preoptica medialis to 43 degrees C resulted in a decrease in food intake despite concomitant hypothermia.
Vernalization, the acceleration of flowering by a long period of cold temperature, ensures that many plants overwinter vegetatively and flower in spring. In Arabidopsis, allelic variation at the FRIGIDA (FRI) locus is a major determinant of natural variation in flowering time. Dominant alleles of FRI confer late flowering, which is reversed to earliness by vernalization. We cloned FRI and analyzed the molecular basis of the allelic variation. Most of the early-flowering ecotypes analyzed carry FRI alleles containing one of two different deletions that disrupt the open reading frame. Loss-of-function mutations at FRI have thus provided the basis for the evolution of many early-flowering ecotypes.
Nanomolar concentrations of peptidoleukotrienes evoke sustained cerebral edema and arterial constriction. Peptidoleukotrienes are thus considered to play an important role in eliciting cerebral edema after cerebral ischemia and vasospasm after subarachnoid hemorrhage. It was hypothesized that the choroid plexus, the locus of the blood-cerebrospinal fluid barrier, might minimize the vasoactivity of locally generated or systemically derived leukotrienes by transporting leukotrienes from cerebrospinal fluid into the blood. Consistent with this hypothesis, leukotriene C4 in vitro was transported into and released from isolated rabbit choroid plexus by a system that was specific, energy-dependent, probenecid-sensitive, and depressed by cold temperatures. The accumulation of leukotriene C4 in the choroid plexus was not dependent on tissue binding or metabolism of leukotriene C4.
Novel strains of obligately chemolithoautotrophic, sulfur-oxidizing bacteria have been isolated from various depths of Lake Fryxell, Antarctica. Physiological, morphological, and phylogenetic analyses showed these strains to be related to mesophilic Thiobacillus species, such as T. thioparus. However, the psychrotolerant Antarctic isolates showed an adaptation to cold temperatures and thus should be active in the nearly freezing waters of the lake. Enumeration by most-probable-number analysis in an oxic, thiosulfate-containing medium revealed that the sulfur-oxidizing chemolithotroph population peaks precisely at the oxycline (9.5 m), although viable cells exist well into the anoxic, sulfidic waters of the lake. The sulfur-oxidizing bacteria described here likely play a key role in the biogeochemical cycling of carbon and sulfur in Lake Fryxell.
Climate change is broadly expected to increase the range of many plant diseases, yet the current status of local thermal adaptation in many pathogens is poorly understood. Xylella fastidiosa (Xf) is a global bacterial plant pathogen that causes Pierce's disease (PD) of grapevines and infects over 700 other host plant species, impacting both agricultural and natural ecosystems. In a common garden experiment with 477 vines in the field, we compared PD outcomes from a local (colder climate in CA) vs non-local (warmer climate in CA) bacterial strain in 13 Mediterranean grapevine varieties over 3 years. Relative to the local strain, there was 77% lower overwinter survival in the non-local strain from a warmer climate, strongly indicating local adaptation in these CA Xf populations. Host genotype also had a significant effect on pathogen winter survival, and grapevine varieties differed in PD susceptibility. Additionally, we assessed in planta evolution of the two pathogen strains over 3 years by whole-genome sequencing 58 field-derived isolates. There were convergent loss-of-function mutations in genes encoding minor Type IV pilin (T4P) proteins, which control twitching motility and other virulence phenotypes, suggesting rapid adaptive evolution. Our results suggest local adaptation to cold temperatures in a bacterial plant pathogen and a possible role for minor Type IV pilins in thermal adaptation. These findings demonstrate the urgent need to incorporate X. fastidiosa evolution and local thermal adaptation into global models of PD spread. Differentiating pathotypes with distinct thermal adaptations will improve disease forecasting and inform quarantine decisions.IMPORTANCEForecasting the movement of plant pathogens is a critical issue under global warming to effectively manage future plant disease outbreaks. Yet, current plant pathogen local thermal adaptation is often unaccounted for, especially in bacterial pathogens. Our study examines local adaptation to temperature in a bacterial plant pathogen, Xylella fastidiosa, that causes disease in grapevines in addition to infecting 700 other plant species. In a large-scale field experiment across 13 grapevine varieties, we demonstrate local adaptation in pathogen winter survival in distinct Xylella fastidiosa strains. Additionally, we found evidence of adaptive evolution in just 3 years, as we observed convergent mutations after resequencing strains that evolved in the field. Our results suggest that X. fastidiosa populations-even within a small geographic area-have distinct adaptations to winter temperatures and may exhibit differential responses to warming winters.
With the application of plate count methods and of the KOH-flexirubin test, bacteria belonging to the Cytophaga group were proved to occur regularly in samples from biological sewage treatment facilities. Generally, the percentage of Cytophaga colonies of the total heterotrophic colonies was lowest in the inflow sewage water as compared with the values found in activated sludge, trickling filter, and effluent samples. During an observation period of 16 months, the highest percentages of cytophagas were found in winter samples from activated sludge and trickling filters. Furthermore, cytophagas were shown to have high percentages of the bacteria lytic to polymeric substrates such as cellulose, chitin, dextran, pectin, xylan, and gelatin. Thus, it is suggested that cytophagas may contribute to sewage purification, especially at cold temperatures and by polymer breakdown. Cytophaga strains isolated were shown to have gliding motility, flexirubin pigmentation, and a low guanine plus cytosine base ratio in common. The strains were roughly subdivided into a spreading, a nonspreading, and a cellulolytic group.
The assembly and budding of human immunodeficiency virus type 1 (HIV-1) at the plasma membrane are directed by the viral core protein Pr55(gag). We have analyzed whether Pr55(gag) has intrinsic affinity for sphingolipid- and cholesterol-enriched raft microdomains at the plasma membrane. Pr55(gag) has previously been reported to associate with Triton X-100-resistant rafts, since both intracellular membranes and virus-like Pr55(gag) particles (VLPs) yield buoyant Pr55(gag) complexes upon Triton X-100 extraction at cold temperatures, a phenotype that is usually considered to indicate association of a protein with rafts. However, we show here that the buoyant density of Triton X-100-treated Pr55(gag) complexes cannot be taken as a proof for raft association of Pr55(gag), since lipid analyses of Triton X-100-treated VLPs demonstrated that the detergent readily solubilizes the bulk of membrane lipids from Pr55(gag). However, Pr55(gag) might nevertheless be a raft-associated protein, since confocal fluorescence microscopy indicated that coalescence of GM1-positive rafts at the cell surface led to copatching of membrane-bound Pr55(gag). Furthermore, extraction of intracellular membranes or VLPs with Brij98 yielded buoyant Pr55(gag) complexes of low density. Lipid analyses of Brij98-treated VLPs suggested that a large fraction of the envelope cholesterol and phospholipids was resistant to Brij98. Collectively, these results suggest that Pr55(gag) localizes to membrane microdomains that are largely resistant to Brij98 but sensitive to Triton X-100, and these membrane domains provide the platform for assembly and budding of Pr55(gag) VLPs.
PRP16 is an RNA-dependent ATPase required for the second catalytic step of splicing in vitro. A dominant suppressor of a branchpoint mutation in Saccharomyces cerevisiae, the prp16-1 allele, contains a Tyr to Asp change in the nucleotide-binding site consensus sequence. We now find that cells harboring the prp16-1 allele have a general growth defect that is exacerbated at cold temperatures. The mutant is dominant over the wild-type gene when overexpressed. Purified Prp16-1 protein binds to the spliceosome with apparently wild-type affinity; however, it only weakly complements the second-step block in a PRP16-depleted extract. Analysis of purified Prp16-1 revealed that the rate of ATP hydrolysis is greatly reduced. These results can account for the dominant negative growth phenotype and argue that the ATPase activity of PRP16 is essential for its role in splicing. Moreover, since PRP16 is a member of the DEAD/H box families, these findings have important implications for a large class of proteins.
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The anterior chambers of 27 rabbit eyes were perfused at constant pressure with room temperature (25 degrees C) or cooled (11 degrees C) balanced salt solution at constant flow rates of 4.8 ml/min or 8.5 ml/min. Intraocular temperature changes in the anterior chamber, anterior vitreous, mid vitreous, and posterior vitreous and on the retina surface were monitored with an intraocular thermocouple probe. Perfusion of the anterior chamber of the pigmented rabbit eye with cooled fluid significantly reduced the temperature of the anterior chamber and anterior vitreous and even that of the retina. Both an increase in the rate of perfusion and a lowering of the perfusion temperature enhanced the cooling effect. The observed decrease in temperatures returned to approximately normal 4 minutes following the cessation of perfusion.