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Respiratory cooling in rattlesnakes.

We used infrared thermography to study respiratory cooling in the rattlesnakes (Viperidae: Crotalinae) and to partition the effects of air temperature, humidity, and activity levels on head-body temperature differences. We observed a single, cooled region centered around the mouth and nasal capsule that extended across the pit membrane at air temperatures above 20 degrees C. Both head and body temperatures of rattlesnakes increased linearly with air temperature. Head-body temperature differentials also increased with air temperature, but declined significantly at higher relative humidities. Rattling rattlesnakes exhibited significantly greater head-body temperature differentials than did resting rattlesnakes. We suggest that respiratory cooling may provide a thermal buffer for the thermoreceptive pit organs at high air temperatures, but caution that this adaptive hypothesis must be tested with direct neural or behavioral assays.

Animals↗

Cutaneous vasculitis: diagnosis and management.

Vasculitis is histologically defined as inflammatory cell infiltration and destruction of blood vessels. Vasculitis is classified as primary (idiopathic, eg, cutaneous leukocytoclastic angiitis, Wegener's granulomatosis) or secondary, a manifestation of connective tissue diseases, infections, adverse drug eruptions, or a paraneoplastic phenomenon. Cutaneous vasculitis, manifested as urticaria, purpura, hemorrhagic vesicles, ulcers, nodules, livedo, infarcts, or digital gangrene, is a frequent and often significant component of many systemic vasculitic syndromes such as lupus or rheumatoid vasculitis and antineutrophil cytoplasmic antibody-associated primary vasculitic syndromes such as Churg-Strauss syndrome. In most instances, cutaneous vasculitis represents a self-limited, single-episode phenomenon, the treatment of which consists of general measures such as leg elevation, warming, avoidance of standing, cold temperatures and tight fitting clothing, and therapy with antihistamines, aspirin, or nonsteroidal anti-inflammatory drugs. More extensive therapy is indicated for symptomatic, recurrent, extensive, and persistent skin disease or coexistence of systemic disease. For mild recurrent or persistent disease, colchicine and dapsone are first-choice agents. Severe cutaneous and systemic disease requires more potent immunosuppression (prednisone plus azathioprine, methotrexate, cyclophosphamide, cyclosporine, or mycophenolate mofetil). In cases of refractory vasculitis, plasmapheresis and intravenous immunoglobulin are viable considerations. The new biologic therapies that work via cytokine blockade or lymphocyte depletion such as tumor alpha inhibitor infliximab and the anti-B-cell antibody rituximab, respectively, are showing benefit in certain settings such as Wegener's granulomatosis, antineutrophil cytoplasmic antibody-associated vasculitis, Behçet's disease, and cryoglobulinemic vasculitis.

Anti-Inflammatory Agents, Non-Steroidal↗

Positioning impact on the surgical patient.

The operating room is unknown territory to most health care providers. It frequently brings up thoughts of blood, strange smells, and cold temperatures. Many nursing programs have scheduled little, if any, time in this environment for students. As a result, few nurses who practice outside of this specialized area understand the patient care events that occur in the operating room. Those who have selected the operating room for their work environment know that it is a somewhat isolated period in the perioperative experience. This article provides insight into this area of patient care and a greater understanding of how patients are positioned, the physiologic impact of these positions, and some consequences that may impact the postoperative care of these patients.

Beds↗

Longevity of cryogenically stored seeds.

Though cryogenic storage is presumed to provide nearly infinite longevity to cells, the actual shelf life achieved under ultra-cold temperatures has not been addressed theoretically or empirically. Here, we report measurable changes in germination of dried seeds stored under liquid nitrogen conditions for >10 years. There was considerable variability in the extent of deterioration among species and accessions within a species. Aging time courses for lettuce seeds stored at temperatures between 50 and -196 degrees C were fit to a form of the Avrami equation to determine rate coefficients and predict half-life of accessions. A reduction in the temperature dependency on aging rate, determined as a break in the Arrhenius plot, occurred at about -15 degrees C, and this resulted in faster deterioration than anticipated from extrapolation of kinetics measured at higher temperatures. The break in Arrhenius behavior occurred at temperatures in between the glass transition temperature (28 degrees C) and the Kauzmann temperature (-42 degrees C) and also coincided with a major triacylglycerol phase change (-40 to -7 degrees C). In spite of the faster than anticipated deterioration, cryogenic storage clearly prolonged shelf life of lettuce seeds with half-lives projected as approximately 500 and approximately 3400 years for fresh lettuce seeds stored in the vapor and liquid phases of liquid nitrogen, respectively. The benefit of low temperature storage (-18 or -135 degrees C) on seed longevity was progressively lost if seeds were first stored at 5 degrees C. Collectively, these results demonstrate that lowering storage temperature progressively increases longevity of seeds. However, cryogenic temperatures were not sufficient to stop deterioration, especially if initial stages of aging were allowed to progress at higher storage temperatures. This work contributes to reliable assessments of the potential benefit and cost of different genebanking strategies.

Cryopreservation↗

Constitutive expression of abiotic stress-inducible hot pepper CaXTH3, which encodes a xyloglucan endotransglucosylase/hydrolase homolog, improves drought and salt tolerance in transgenic Arabidopsis plants.

Xyloglucan endotransglucosylase/hydrolase (XTH) has been recognized as a cell wall-modifying enzyme, participating in the diverse physiological roles. From water-stressed hot pepper plants, we isolated three different cDNA clones (pCaXTH1, pCaXTH2, and pCaXTH3) that encode XTH homologs. RT-PCR analysis showed that three CaXTH mRNAs were concomitantly induced by a broad spectrum of abiotic stresses, including drought, high salinity and cold temperature, and in response to stress hormone ethylene, suggesting their role in the early events in the abiotic-related defense response. Transgenic Arabidopsis plants that constitutively expressed the CaXTH3 gene under the control of the CaMV 35S promoter exhibited abnormal leaf morphology; the transgenic leaves showed variable degrees of twisting and bending along the edges, resulting in a severely wrinkled leaf shape. Microscopic analysis showed that 35S-CaXTH3 leaves had increased numbers of small-sized cells, resulting in disordered, highly populated mesophyll cells in each dorsoventral layer, and appeared to contain a limited amount of starch. In addition, the 35S-CaXTH3 transgenic plants displayed markedly improved tolerance to severe water deficit, and to lesser extent to high salinity in comparison with the wild-type plants. These results indicate that CaXTH3 is functional in heterologous Arabidopsis cells, thereby effectively altering cell growth and also the response to abiotic stresses. Although the physiological function of CaXTHs is not yet clear, there are several possibilities for their involvement in a subset of physiological responses to counteract dehydration and high salinity stresses in transgenic Arabidopsis plants.

Amino Acid Sequence↗

Validated HPLC analytical method with programmed wavelength UV detection for simultaneous determination of DRF-4367 and Phenol red in rat in situ intestinal perfusion study.

A simple, precise and accurate isocratic reverse-phase liquid chromatography method with programmed wavelength detection has been validated to quantify DRF-4367 and Phenol red, simultaneously for application in rat in situ single pass intestinal perfusion study to assess intestinal permeability of DRF-4367, a novel cox-2 inhibitor. The method was validated on RP C-18 analytical column. Mobile phase consisted of sodium dihydrogen orthophosphate (pH 3.2, 0.01 M)-acetonitrile-methanol (30:50:20, v/v/v). The developed method has a short run time of 12 min with a flow rate of 1.0 ml/min. The injector volume was set to 20 microl and acquisition was carried out using a PDA detector while processing was done by timed wavelength extraction. The percentage R.S.D. and recovery in all studies indicated that the method was suitable for the intended purpose. The validated method was found to be linear and precise in the working range. Suitability of storage at cold temperature was ensured along with complete sample recovery.

Animals↗

High-flow endobronchial cooled humidified air protects non-heart-beating donor rat lungs against warm ischemia.

OBJECTIVE: Lungs from non-heart-beating donors for transplantation require protection against warm ischemic damage. Minimally invasive techniques are required to reduce organ damage during the warm ischemic period because invasive surgical procedures are often not feasible at this time. This study investigated the preservative effect of high-flow endobronchial cooled humidified air during warm ischemia in non-heart-beating donor rat lungs. METHODS: Fourteen animals were divided into a Cooling group (n = 7), which received cooled air/saline spray during a 2-hour warm ischemic period, and a Control group (n = 7), which received no cooling. After ischemia the lungs were reperfused on an isolated lung perfusion apparatus. RESULTS: Endobronchial temperatures in the Cooling and Control groups were 8 degrees C and 36 degrees C at 10 minutes, and 5 degrees C and 35 degrees C at 20 minutes, respectively (P < .0001). Lung core and surface temperatures in the Cooling group were also lower than those in the corresponding Control group (P < .0001). After reperfusion, pulmonary arterial pressure (P = .003) and peak airway pressure (P = .002) were lower in the Cooling group than in the Control group. Higher pulmonary venous PO2 (P = .02), higher adenosine triphosphate levels (P = .01), and lower wet/dry lung weight ratio (P = .003) were seen in the Cooling group compared with the Control group. CONCLUSIONS: High-flow endobronchial cooled humidified air can decrease lung temperature and improve post-ischemic pulmonary function and adenosine triphosphate levels in non-heart-beating donor lungs.

Adenosine Triphosphatases↗

Effect of material properties of composite restoration on the strength of the restoration-dentine interface due to polymerization shrinkage, thermal and occlusal loading.

The purpose of this investigation was to adopt an analytical approach to analyse stresses at the restoration-dentine interface caused by polymerization shrinkage, occlusal and thermal loading with the primary focus on evaluating the effect of the material properties of the composite restoration on the strength of the interface. Some essential simplifications were employed to derive an explicit analytical solution. The results confirm previous findings that interfacial stresses due to polymerization shrinkage are increased with the higher modulus of elasticity of the restoration, while Poisson's ratio of the restorative material has a very small influence on these stresses. Occlusal loading resulted in much lower interfacial stresses when compared to shrinkage and thermal loads. The obtained results were in a good agreement with other numerical and clinical studies. From the modelling analysis it was found that the majority of commercially available composite restorative materials are expected to create significant interfacial stresses when subjected to cold temperatures. In addition, it was shown that there is a considerable potential for interfacial stresses to be minimised by an appropriate selection of thermo-mechanical properties of the restorative material especially with the new finding on the negative temperature variation effect.

Adhesiveness↗

Enhanced thermal avoidance in mice lacking the ATP receptor P2X3.

P2X3 is an ATP-gated cation channel subtype expressed by a subpopulation of primary sensory neurons. In vivo spinal cord recordings in mice lacking P2X3 (P2X3-/-) have suggested that this protein may be important for the coding of peripheral warm stimuli. To explore this possibility more thoroughly, we examined behavioral and electrophysiological responses to thermal stimuli in P2X3-/- mice. As previously reported, recording from the spinal cord dorsal horn of anesthetized P2X3-/- mice revealed a blunted response of wide dynamic range neurons to hind paw heating. When placed in a thermal gradient, however, P2X3-/- mice exhibited an unexpectedly enhanced avoidance of both hot and cold temperatures, relative to controls. In the tail immersion test, mutant mice exhibited shorter withdrawal latencies at temperatures above and below thermoneutrality. Consistent with these changes, P2X3-/- mice exhibited enhanced induction of spinal cord c-FOS following hind paw heating to 45 degrees C. Thus, gain- and loss-of-function thermosensory phenotypes coexist in P2X3-/- mice. No changes in thermal preference were observed in wild-type mice injected subcutaneously with the P2X3 antagonist, A317491 or intrathecally with the P2X3 and P2X1 antagonist TNP-ATP. The reason for this apparent discrepancy is unclear, but we cannot exclude the possibility that compensatory events contribute, at least in part, to the P2X3-/- phenotype. Regardless, this study illustrates the utility of thermal preference assays as part of a comprehensive approach to the analysis of mouse thermosensation.

Analysis of Variance↗

Dynamics of solutes and dissolved oxygen in shallow urban groundwater below a stormwater infiltration basin.

Artificial recharge of urban aquifers with stormwater has been used extensively in urban areas to dispose of stormwater and compensate for reduced groundwater recharge. However, stormwater-derived sediments accumulating in infiltration beds may act as a source of dissolved contaminants for groundwater. Concentrations of hydrocarbons, heavy metals, nutrients and dissolved oxygen (DO) were monitored at multiple depths in shallow groundwater below a stormwater infiltration basin retaining large amounts of contaminated organic sediments. Multilevel wells and multiparameter loggers were used to examine changes in groundwater chemistry occurring over small spatial and temporal scales. Rainfall events produced a plume of low-salinity stormwater in the first 2 m below the groundwater table, thereby generating steep vertical physico-chemical gradients that resorbed during dry weather. Heavy metals and hydrocarbons were below reference concentrations in groundwater and aquifer sediments, indicating that they remained adsorbed onto the bed sediments. However, mineralization of organic sediments was the most probable cause of elevated concentrations of phosphate and DOC in groundwater. DO supply in groundwater was severely limited by bed respiration which increased with temperature. Cold winter stormwater slightly re-oxygenated groundwater, whereas warm summer stormwater lowered DO concentrations in groundwater. Among several results provided by this study, it is recommended for management purposes that infiltration practices should minimize the contact between inflow stormwater and organic sediments retained in infiltration basins.

Journal Article↗

Method for detecting classes of microcystins by combination of protein phosphatase inhibition assay and ELISA: comparison with LC-MS.

Depending on the class of microcystin the protein phosphatase inhibition assay shows different sensitivities to different classes of toxin. We have determined that the IC50 values obtained from dose-response curves for the inhibition of the enzyme by micro-cystin LR, nodularin, YR, and RR were 2.2, 1.8, 9 and 175 nM, respectively. When equimolar amounts of these toxins were determined by the ELISA assay with microcystin LR as the standard, the assay showed equivalence in toxin responses. However, when the toxins were determined by the protein phosphatase inhibition assay using microcystin LR as the standard, the ratios of the values determined by PP-2A to ELISA decreased in the order: nodularin (2.23) microcystin LR (1.1)> microcystin YR (0.63)> microcystin RR (0.06). When the ratios for each standard were plotted against the IC50 values, the log-log plot was negative linear, and the lowest value for the IC50 corresponded with the lowest ratio. The differential sensitivity of the PP-2A assay to the various standards was used to establish an indicative toxicity ranking (ITR) where a ranking of 1 (the highest) was assigned to ratios of > or = 0.8 or greater, and 3 (the lowest) to values < or = 0.2. The three ranking classes corresponded to toxin equivalence represented by the four standards. The new method allows not only the determination of microcystin toxins in terms of stoichiometry (ELISA) but also in terms of indicative toxicity. The method can be performed using the same instrument (e.g. multiwell fluorimeter with absorbance capability) and offers an advantage to methods presently used to determine microcystins (e.g. ELISA or LC-MS). The former has the propensity to overestimate toxicity because it measures equivalence to microcystin LR and is a stoichiometric measurement and the latter has the disadvantage in that relatively few of the microcystins that occur naturally are available as standards. The new method was applied to the analysis of sample from lakes and streams from temperate locations and to extracts of cyanobacterial mats from ponds and streams in cold temperature locations.

Bacterial Toxins↗

Genomic signatures of the Arctic-adapted North American gray wolf ecotype.

The Arctic Circle is one of Earth's most extreme environments. It features cold temperatures, resource shortages, and near-complete winter darkness. Here, we generated a chromosome-level genome assembly of a male wolf from the Arctic Archipelago (Canis lupus arctos). Our assembly and that of the related C. l. orion from Greenland was used to identify candidate genic and regulatory features of Arctic-adapted polar wolves, ranging from selection acting on standing variation and amino acid changes in genes to conserved non-exonic elements (CNEEs) that may regulate gene expression. We identified genes and nearby CNEEs associated with thermoregulation (e.g., APOB), coat color and patterning (e.g., GOLGB1), and DNA damage response (e.g., POLQ). In vitro assays supported changes in polar wolf gene (POLQ and TRPV2 amino acid substitutions) and CNEE function. Our report offers insights into the genetic mechanisms underlying polar wolf adaptations, laying a foundation for future studies on Arctic canines.

Amino acid substitutions↗

Tubulin polymerizing activity of dictyostatin-1, a polyketide of marine sponge origin.

Dictyostatin-1 had previously been isolated from a marine sponge of the genus Spongia sp. and described as a cytotoxic agent to murine and human cancer cells, but its mechanism of activity was unknown. In a routine screening assay used to detect cytotoxic compounds of marine origin, dictyostatin-1 was identified as a highly active component in an extract from a Lithistida sponge and exploration into its pharmacology was pursued. Initial studies demonstrated that dictyostatin-1 arrested cells in the G(2)/M phase of the cell cycle. Staining of these cells with antitubulin revealed cells having multiple aster formations and microtubule matrix bundling patterns similar to that seen in cells exposed to paclitaxel. Dictyostatin-1 was able to induce the polymerization of purified bovine brain tubulin in vitro and the polymerized tubulin remained stable at cold temperatures. Dictyostatin-1 also proved to be highly potent in two paclitaxel-resistant human cancer cell lines expressing active P-glycoprotein. Together, these results indicate that dictyostatin-1 is a potent inducer of tubulin polymerization and retains activity in cells expressing the P-glycoprotein efflux pump.

Animals↗

Cooling lesions of the lateral parabrachial nucleus during LiCl activation block acquisition of conditioned taste avoidance in male rats.

Lesions of the lateral parabrachial nucleus (lPBN) disrupt acquisition of LiCl-induced conditioned taste avoidance. Animals with lesions in this area also fail to exhibit taste neophobia. This raises the possibility that an inability of rats to recognize the taste solution as novel contributes to the deficit in taste avoidance learning. If this is the case, then one would expect conditioned taste avoidance not to be disrupted if the lPBN is functional during taste processing but not during LiCl processing. The first three experiments demonstrated that cooling was a viable method by which to temporarily inactivate the lPBN. Measurement of neural temperature during cooling indicated that the lPBN was cooled to temperatures that have been shown to block synaptic transmission but not axonal transmission. Cooling the lPBN itself induced a conditioned avoidance to a sucrose solution but this avoidance was abolished by exposure to daily cooling for 1 week prior to acquisition. In experiment 4, all animals were preexposed to lPBN cooling for 1 week. Those rats that received cooling lesions during a period that started immediately after sucrose solution consumption and extended through the peak effectiveness of LiCl failed to acquire a taste avoidance. These results fail to support the hypothesis that the deficit in taste avoidance learning after permanent lesions of the lPBN is due to an inability of lesioned animals to recognize the taste as novel. They are consistent with the hypothesis that this neural area processes ascending information about LiCl.

Animals↗

Development of a two-dimensional liquid chromatography system with trapping and sample enrichment capabilities.

A two-dimensional HPLC system was developed where "heart-cutting" chromatography, in conjunction with cold temperature trapping, was used to isolate and concentrate specific sample analytes. Low molecular mass polystyrene oligomers were used as model compounds to illustrate the operation of the instrument and evaluate the performance of the trapping system. A critical factor in the operation of the trapping system was the relative degree of retention between the first column and the trapping column. The results of this study showed that up to 32 consecutive heart-cut fractions from the first separation dimension could be stored in a trapping column with good analyte recovery and without significant loss in resolution upon elution on the second separation dimension.

Chromatography, High Pressure Liquid↗

An in vivo recording of variations in oral temperature during meals: a pilot study.

Thermocycling is often used in the in vitro evaluation of marginal leakage of dental restorations. The in vivo temperatures occurring during meals were recorded at different surfaces of restored teeth, to compare the variations in temperature with laboratory thermocycling. Temperatures were recorded with thermocouples at three different sites: (1) on the facial surface of a silver amalgam restoration, (2) at the base of a silver amalgam restoration and (3) within the root canal. The maximal temperature differences between upper and lower extremes were 29.6 degrees C at the base of a coronal restoration, 27.1 degrees C at the facial surface, and 11.8 degrees C within the root canal. A practical regimen for these experiments is suggested in the light of these ranges because many thermocycling regimens in in vitro studies appear extreme or unrealistic.

Beverages↗

Temperature rise in pulpal chamber during fabrication of provisional resinous crowns.

STATEMENT OF PROBLEM: The heat generated during the exothermic polymerization reaction of autopolymerizing resinous materials and the heat generated by ultraviolet lamps during irradiation of photopolymerizing resinous materials could cause pulpal damage when a direct technique is used to fabricate provisional restorations. This could occur if temperature elevations overcome the physiological heat dissipating mechanisms of the dental-periodontal system. PURPOSE: This in vitro study compared the rise in temperatures in the pulpal chamber during fabrication of provisional complete veneer crowns by direct method with different autopolymerizing and photopolymerizing resins. The effect of curing resinous crowns in different matrices, such as a polyvinyl siloxane impression and a vaccuum-formed polypropylene sheet, was also evaluated. RESULTS: The results demonstrated that the amount of heat generated during resin polymerization and transmitted to the pulpal chamber could be damaging to pulpal tissues including odontoblasts. When curing of provisional resinous crowns was performed in the polyvinyl siloxane impression, significantly lower temperatures were recorded compared with curing in the vacuum-formed polypropylene sheet. CONCLUSIONS: To prevent pulpal damage, effective cooling procedures are strongly recommended when directly fabricating resinous provisional crowns.

Analysis of Variance↗