Periodic Paralysis in a Child With Thermosensitive Mitochondrial Trifunctional Protein Deficiency.

Mitochondrial trifunctional protein (MTP) deficiency is a fatty acid oxidation disorder associated with a spectrum of phenotypes. Patients with high residual enzyme activity tend to have milder phenotypes, and recently, fever-induced episodic myopathy was reported in association with a thermosensitive form of MTP deficiency. We report a 10-year-old male with recurrent episodes of acute flaccid paralysis involving upper and lower extremities in association with bulbar muscle weakness in the context of febrile illness, a phenotype reminiscent of recurrent periodic paralysis. The episodes started at the age of 3 years and have always been followed by full recovery within 1-2 weeks with no residual weakness. Whole exome sequencing revealed a homozygous c.2132C > T, p.(Pro711Leu) variant in HADHA. The variant leads to mildly reduced long-chain hydroxyacyl-CoA dehydrogenase (LCHAD) and long-chain ketoacyl-CoA thiolase (LCKAT) enzyme activities and reduced MTP protein expression in patient's fibroblasts when cultured at 37°C. Enzyme activities and MTP protein expression diminished when fibroblasts were cultured at 40°C. This is the first published report of confirmed recurrent periodic paralysis as a manifestation of a thermosensitive form of MTP deficiency, and it calls for this condition to be considered when evaluating patients with recurrent periodic paralysis given therapeutic implications.

Screening methods for thermotolerance in pollen.

Plant reproduction is highly susceptible to temperature stress. The development of the male gametophyte in particular represents a critical element in the reproductive cycle with high sensitivity to elevated temperatures. Various methods have been used to test the effect of temperature stress on pollen performance or to determine the degree of susceptibility of given species and genotypes. The information gained informs the development of new crop varieties suited to grow under warmer conditions arising through climate change and facilitates predicting the behavior of natural populations under these conditions. The characterization of pollen performance typically employs the terms pollen viability and pollen vigor, which, however, are not necessarily used consistently across studies. Pollen viability is a nominal parameter and is often assayed relying on cellular features as proxy to infer the capability of pollen grains to germinate and complete double fertilization. Alternatively, pollen germination can be determined through in vitro growth assays, or by monitoring the ability of pollen tubes to complete different progamic steps in vivo (ability to reach an ovule, release sperm cells, lead to seed set). Pollen vigor is an ordinal parameter that describes pollen tube growth rate or the efficiency of pollen tube growth as inferred by its morphology or growth pattern. To ensure consistent and relevant terminology, this review defines these terms and summarizes the methodologies used to assess them.

Temperature-Dependent Activation of Thermosensitive Transient Receptor Potential Channels.

Temperature detection is essential for the survival and perpetuation of any species. Thermoreceptors in the skin sense the body temperature and also the temperatures of the ambient air and the objects. In 1997, Dr. David Julius and his colleagues found that a receptor expressed in small-diameter primary sensory neurons was activated by capsaicin (the pungent chemical in hot pepper). This receptor was also activated by temperature above 42 °C. That was the first time that a thermal receptor in primary sensory neurons has been identified. This receptor is named transient receptor potential vanilloid 1 (TRPV1). Now, 11 thermosensitive TRP channels are known. In this chapter, we summarize the reports and analyze thermosensitive TRP channels in a variety of ways to clarify the activation mechanisms by which temperature changes are sensed.

Towards a central origin of nociceptive hypersensitivity in adult rats after a neonatal maternal separation.

Early life adversities influence a nervous system still in development with long-term consequences for later life. These include nociceptive circuit alterations critical to shape an adaptive pain response to protect the organism from potential damage. Adult rats with a history of neonatal maternal separation (NMS) display visceral and somatic nociceptive hypersensitivity and inefficient analgesic responses to stress. In this study, we have characterized the consequences of NMS on wide dynamic range neurons (WDR) in the spinal cord of anaesthetized adult rats during the nociceptive processing of hot and cold noxious information. We found that WDR neurons of NMS rats display an excessive coding of mechanical and thermal information applied at the rat's hindpaws. This nicely explains the hypernociceptive behaviours seen after noxious mechanical, cold and hot peripheral stimulation. A peripheral change in the expression of molecular transducers for these stimuli (i.e., TRPV1, TRPM8 and TRPA1) does not seem to account for this general hyperexcitability. Instead, a decreased chloride-mediated inhibitory tone on WDR neurons may play a role as indicated by the abnormal elevation of the type 1 Na-K-Cl cotransporter transcripts. Altogether, we propose that long-term consequences of NMS are associated with reduced spinal cord inhibition favouring the expression of pain hypersensitivity. We cannot exclude that this phenomenon is also present at supraspinal sites, as other NMS-associated symptoms include excessive anxiety and impaired sociability.

The Contribution of TRPA1 to Corneal Thermosensitivity and Blink Regulation in Young and Aged Mice.

The role of TRPA1 in the thermosensitivity of the corneal cold thermoreceptor nerve endings was studied in young and aged mice. The contribution of the TRPA1-dependent activity to basal tearing and thermally-evoked blink was also explored. The corneal cold thermoreceptors' activity was recorded extracellularly in young (5-month-old) and aged (18-month-old) C57BL/6WT (WT) and TRPA1-/- knockout (TRPA1-KO) mice at basal temperature (34 °C) and during cooling (15 °C) and heating (45 °C) ramps. The blink response to cold and heat stimulation of the ocular surface and the basal tearing rate were also measured in young animals using orbicularis oculi muscle electromyography (OOemg) and phenol red threads, respectively. The background activity at 34 °C and the cooling- and heating-evoked responses of the cold thermoreceptors were similar in WT and TRPA1-KO animals, no matter the age. Similar to the aged WT mice, in the young and aged TRPA1-KO mice, most of the cold thermoreceptors presented low frequency background activity, a low cooling threshold, and a sluggish response to heating. The amplitude and duration of the OOemg signals correlated with the magnitude of the induced thermal change in the WT but not in the TRPA1-KO mice. The basal tearing was similar in the TRPA1-KO and WT mice. The electrophysiological data suggest that the TRPA1-dependent nerve activity, which declines with age, contributes to detecting the warming of the ocular surface and also to integrating the thermally-evoked reflex blink.

TFIIH stabilization recovers the DNA repair and transcription dysfunctions in thermo-sensitive trichothiodystrophy.

Trichothiodystrophy (TTD) is a rare hereditary disease whose prominent feature is brittle hair. Additional clinical signs are physical and neurodevelopmental abnormalities and in about half of the cases hypersensitivity to UV radiation. The photosensitive form of TTD (PS-TTD) is most commonly caused by mutations in the ERCC2/XPD gene encoding a subunit of the transcription/DNA repair complex TFIIH. Here we report novel ERCC2/XPD mutations affecting proper protein folding, which generate thermo-labile forms of XPD associated with thermo-sensitive phenotypes characterized by reversible aggravation of TTD clinical signs during episodes of fever. In patient cells, the newly identified XPD variants result in thermo-instability of the whole TFIIH complex and consequent temperature-dependent defects in DNA repair and transcription. Improving the protein folding process by exposing patient cells to low temperature or to the chemical chaperone glycerol allowed rescue of TFIIH thermo-instability and a concomitant recovery of the complex activities. Besides providing a rationale for the peculiar thermo-sensitive clinical features of these new cases, the present findings demonstrate how variations in the cellular concentration of mutated TFIIH impact the cellular functions of the complex and underlie how both quantitative and qualitative TFIIH alterations contribute to TTD clinical features.

Tilapia prolactin cells are thermosensitive osmoreceptors.

Prolactin (PRL) cells within the rostral pars distalis (RPD) of euryhaline and eurythermal Mozambique tilapia, Oreochromis mossambicus, rapidly respond to a hyposmotic stimulus by releasing two distinct PRL isoforms, PRL188 and PRL177. Here, we describe how environmentally relevant temperature changes affected mRNA levels of prl188 and prl177 and the release of immunoreactive prolactins from RPDs and dispersed PRL cells. When applied under isosmotic conditions (330 mosmol/kgH2O), a 6°C rise in temperature stimulated the release of PRL188 and PRL177 from both RPDs and dispersed PRL cells under perifusion. When exposed to this same change in temperature, ∼50% of dispersed PRL cells gradually increased in volume by ∼8%, a response partially inhibited by the water channel blocker, mercuric chloride. Following their response to increased temperature, PRL cells remained responsive to a hyposmotic stimulus (280 mosmol/kgH2O). The mRNA expression of transient potential vanilloid 4, a Ca2+-channel involved in hyposmotically induced PRL release, was elevated in response to a rise in temperature in dispersed PRL cells and RPDs at 6 and 24 h, respectively; prl188 and prl177 mRNAs were unaffected. Our findings indicate that thermosensitive PRL release is mediated, at least partially, through a cell-volume-dependent pathway similar to how osmoreceptive PRL release is achieved.

Thermo-sensitive mitochondrial trifunctional protein deficiency presenting with episodic myopathy.

Mitochondrial trifunctional protein (MTP) is involved in long-chain fatty acid ß-oxidation (lcFAO). Deficiency of one or more of the enzyme activities as catalyzed by MTP causes generalized MTP deficiency (MTPD), long-chain hydroxyacyl-CoA dehydrogenase deficiency (LCHADD), or long-chain ketoacyl-CoA thiolase deficiency (LCKATD). When genetic variants result in thermo-sensitive enzymes, increased body temperature (e.g. fever) can reduce enzyme activity and be a risk factor for clinical decompensation. This is the first description of five patients with a thermo-sensitive MTP deficiency. Clinical and genetic information was obtained from clinical files. Measurement of LCHAD and LCKAT activities, lcFAO-flux studies and palmitate loading tests were performed in skin fibroblasts cultured at 37°C and 40°C. In all patients (four MTPD, one LCKATD), disease manifested during childhood (manifestation age: 2-10 years) with myopathic symptoms triggered by fever or exercise. In four patients, signs of retinopathy or neuropathy were present. Plasma long-chain acylcarnitines were normal or slightly increased. HADHB variants were identified (at age: 6-18 years) by whole exome sequencing or gene panel analyses. At 37°C, LCHAD and LCKAT activities were mildly impaired and lcFAO-fluxes were normal. Remarkably, enzyme activities and lcFAO-fluxes were markedly diminished at 40°C. Preventive (dietary) measures improved symptoms for most. In conclusion, all patients with thermo-sensitive MTP deficiency had a long diagnostic trajectory and both genetic and enzymatic testing were required for diagnosis. The frequent absence of characteristic acylcarnitine abnormalities poses a risk for a diagnostic delay. Given the positive treatment effects, upfront genetic screening may be beneficial to enhance early recognition.

Incubating green turtle (Chelonia mydas) eggs at constant temperatures: Hatching success, hatchling morphology and post-hatch growth.

Past studies applying constant-temperature incubation of eggs have involved all species of sea turtles, but rarely can we find a single one incubating eggs at three or more temperatures. Here, we incubated green turtle (Chelonia mydas) eggs from Ganquan Island, South China Sea, at five constant temperatures (26, 28, 30, 32 and 34 °C) to determine hatching success, incubation length and hatchling phenotype at each test temperature and temperatures optimal for embryonic development. Temperature affected hatching success, incubation length and all seven examined hatchlings traits, and clutch origin affected three (head length, fore-flipper length and hind-flipper length) of the seven. Hatching success was lowest at 34 °C and none of hatchlings hatched at this temperature was normal and survived over one week. The rate of embryonic development and the rate of post-hatch growth both were lowest at 26 °C. Given that low survival and growth rates can translate into reduced individual fitness, we conclude that both 26 °C and 34 °C are unsuitable for incubation of C. mydas eggs. Post-hatch growth was fastest in hatchlings incubated at 30 °C, and eggs of C. mydas incubated at temperatures around 30 °C are more likely to produce mixed sexes. Accordingly, we conclude that temperatures within the range from 28 °C to 32 °C are generally optimal for embryonic development of C. mydas.

Deletion of Yersinia pestis ail Causes Temperature-Sensitive Pleiotropic Effects, Including Cell Lysis, That Are Suppressed by Carbon Source, Cations, or Loss of Phospholipase A Activity.

Maintenance of phospholipid (PL) and lipopoly- or lipooligosaccharide (LPS or LOS) asymmetry in the outer membrane (OM) of Gram-negative bacteria is essential but poorly understood. The Yersinia pestis OM Ail protein was required to maintain lipid homeostasis and cell integrity at elevated temperature (37°C). Loss of this protein had pleiotropic effects. A Y. pestis Δail mutant and KIM6+ wild type were systematically compared for (i) growth requirements at 37°C, (ii) cell structure, (iii) antibiotic and detergent sensitivity, (iv) proteins released into supernatants, (v) induction of the heat shock response, and (vi) physiological and genetic suppressors that restored the wild-type phenotype. The Δail mutant grew normally at 28°C but lysed at 37°C when it entered stationary phase, as shown by cell count, SDS-PAGE of cell supernatants, and electron microscopy. Immunofluorescence microscopy showed that the Δail mutant did not assemble Caf1 capsule. Expression of heat shock promoter rpoE or rpoH fused to a lux operon reporter were not induced when the Δail mutant was shifted from 28°C to 37°C (P < 0.001 and P < 0.01, respectively). Mutant lysis was suppressed by addition of 11 mM glucose, 22 or 44 mM glycerol, 2.5 mM Ca2+, or 2.5 mM Mg2+ to the growth medium or by a mutation in the phospholipase A gene (pldA::miniTn5, ΔpldA, or PldAS164A). A model accounting for the temperature-sensitive lysis of the Δail mutant and the Ail-dependent stabilization of the OM tetraacylated LOS at 37°C is presented. IMPORTANCE The Gram-negative pathogen Yersinia pestis transitions between a flea vector (ambient temperature) and a mammalian host (37°C). In response to 37°C, Y. pestis modifies its outer membrane (OM) by reducing the fatty acid content in lipid A, changing the outer leaflet from being predominantly hexaacylated to being predominantly tetraacylated. It also increases the Ail concentration, so it becomes the most prominent OM protein. Both measures are needed for Y. pestis to evade the host innate immune response. Deletion of ail destabilizes the OM at 37°C, causing the cells to lyse. These results show that a protein is essential for maintaining lipid asymmetry and lipid homeostasis in the bacterial OM.

Thermosensitivity of the voltage-dependent activation of calcium homeostasis modulator 1 (calhm1) ion channel.

Calcium homeostasis modulator 1 (calhm1) proteins form an outwardly rectifying nonselective ion channel having exceedingly slow kinetics and low sensitivity to voltage that is shifted by lowering extracellular Ca2+ ([Ca2+]e). Here we found that physiological temperature dramatically facilitates the voltage-dependent activation of the calhm1 current (Icalhm1); increased amplitude (Q10, 7-15) and fastened speed of activation. Also, the leftward shift of the half-activation voltage (V1/2) was similary observed in the normal and lower [Ca2+]e. Since calhm1 is highly expressed in the brain and taste cells, the thermosensitivity should be considered in their electrophysiology.

Synthesis and characterization of a thermosensitive solid amine biomass adsorbent for carbon dioxide adsorption.

A thermosensitive solid amine fiber SF-AM-co-NIPAM-HBP-NH2 was synthesized by grafting temperature-sensitive monomer N-isopropyl acrylamide (NIPAM) as well as acrylamide (AM) onto the surface of substrate sisal fiber, and further aminating with hyperbranched amine. FTIR, 13C NMR, SEM, EA and TGA were used to confirm the structure and chemical properties of the grafted fibers. Swelling ratio and CO2 adsorption-desorption experiment were investigated to verify the thermo-sensitivity of the grafted fibers and their CO2 adsorption-desorption behavior. Compared with conventional solid amine adsorbents regenerated around 140 °C, SF-AM-co-NIPAM-HBP-NH2 (1:1) with NIPAM could be regenerated at a much lower temperature of 60 °C, while still maintain a high CO2 adsorption capacity (2.61 mmol/g), comparable to that of SF-AM-HBP-NH2 (2.73 mmol/g) before NIPAM introduction. Its excellent regeneration property and the effect of energy consumption reduction make it possible to be used for CO2 adsorption in industrial process.

The cyclin-dependent kinase G group defines a thermo-sensitive alternative splicing circuit modulating the expression of Arabidopsis ATU2AF65A.

The ability to adapt growth and development to temperature variations is crucial to generate plant varieties resilient to predicted temperature changes. However, the mechanisms underlying plant response to progressive increases in temperature have just started to be elucidated. Here, we report that the cyclin-dependent kinase G1 (CDKG1) is a central element in a thermo-sensitive mRNA splicing cascade that transduces changes in ambient temperature into differential expression of the fundamental spliceosome component, ATU2AF65A. CDKG1 is alternatively spliced in a temperature-dependent manner. We found that this process is partly dependent on both the cyclin-dependent kinase G2 (CDKG2) and the interacting co-factor CYCLIN L1 (CYCL1), resulting in two distinct messenger RNAs. The relative abundance of both CDKG1 transcripts correlates with ambient temperature and possibly with different expression levels of the associated protein isoforms. Both CDKG1 alternative transcripts are necessary to fully complement the expression of ATU2AF65A across the temperature range. Our data support a previously unidentified temperature-dependent mechanism based on the alternative splicing (AS) of CDKG1 and regulated by CDKG2 and CYCL1. We propose that changes in ambient temperature affect the relative abundance of CDKG1 transcripts, and this in turn translates into differential CDKG1 protein expression coordinating the AS of ATU2AF65A.

Defects in mating behavior and tail morphology are the primary cause of sterility in Caenorhabditiselegans males at high temperature.

Reproduction is a fundamental imperative of all forms of life. For all the advantages sexual reproduction confers, it has a deeply conserved flaw: it is temperature sensitive. As temperatures rise, fertility decreases. Across species, male fertility is particularly sensitive to elevated temperature. Previously, we have shown in the model nematode Caenorhabditiselegans that all males are fertile at 20°C, but almost all males have lost fertility at 27°C. Male fertility is dependent on the production of functional sperm, successful mating and transfer of sperm, and successful fertilization post-mating. To determine how male fertility is impacted by elevated temperature, we analyzed these aspects of male reproduction at 27°C in three wild-type strains of C. elegans: JU1171, LKC34 and N2. We found no effect of elevated temperature on the number of immature non-motile spermatids formed. There was only a weak effect of elevated temperature on sperm activation. In stark contrast, there was a strong effect of elevated temperature on male mating behavior, male tail morphology and sperm transfer such that males very rarely completed mating successfully when exposed to 27°C. Therefore, we propose a model where elevated temperature reduces male fertility as a result of the negative impacts of temperature on the somatic tissues necessary for mating. Loss of successful mating at elevated temperature overrides any effects that temperature may have on the germline or sperm cells.

Electrophysiological properties of thermosensitive neurons in slices of rat lateral parabrachial nucleus.

Both warm- and cold-sensitive neurons are found in the lateral parabrachial nucleus (LPB), a crucial relay for skin temperature information from the spinal cord to the preoptic area. The aims of this study were to investigate the electrophysiological properties of temperature-sensitive and -insensitive neurons in brain slices, and elucidate the basic mechanisms underlying the thermosensitivity of rat LPB neurons. In warm-sensitive neurons, temperature exerted no significant effects on resting membrane potential (RMP), threshold potential, and amplitude of the afterhyperpolarizing potential. However, warming significantly increased the prepotential rates of depolarization and the inactivation rates of potassium A current (IA) in warm-sensitive neurons, which in turn shortened their interspike interval and elevated the firing rate. In contrast, temperature had no significant effects on the depolarizing prepotentials and inactivation rate of IA in temperature-insensitive neurons. Besides, in cold-sensitive neurons, cooling and warming produced membrane depolarization and hyperpolarization, respectively, and there was a strong correlation between firing rate and membrane potential thermosensitivity. Nevertheless, temperature exhibited no significant effect on the depolarizing prepotential of cold-sensitive neurons. These results suggest that LPB neuronal warm sensitivity may reside in the temperature-dependent prepotentials and IA, while neuronal cold sensitivity might be mainly due to heat-induced changes in RMP.

Mechano- and thermosensitivity of injured muscle afferents 20 to 80 days after nerve injury.

Chronic injury of limb nerves leading to neuropathic pain affects deep somatic nerves. Here the functional properties of injured afferent fibers in the lateral gastrocnemius-soleus nerve were investigated 20 and 80 days after suturing the central stump of this muscle nerve to the distal stump of the sural nerve in anesthetized rats. Neurophysiological recordings were made from afferent axons identified in either the sciatic nerve (87 A-, 63 C-fibers) or the dorsal root L4/L5 (52 A-, 26 C-fibers) by electrical stimulation of the injured nerve. About 70% of the functionally identified A-fibers had regenerated into skin by 80 days after nerve suture; the remaining A-fibers could be activated only from the injured nerve. In contrast, 93% of the functionally identified C-fibers could only be activated from the injured sural nerve after 80 days. Nearly half of the injured A- (45%) and C-fibers (44%) exhibited ongoing and/or mechanically or thermally evoked activity. Because ~50% of the A- and C-fibers are somatomotor or sympathetic postganglionic axons, respectively, probably all injured muscle afferent A- and C-fibers developed ectopic activity. Ongoing activity was present in 17% of the A- and 46% of the C-fibers. Mechanosensitivity was present in most injured A- (99%) and C-fibers (85%), whereas thermosensitivity was more common in C-fibers (cold 46%, heat 47%) than in A-fibers (cold 18%, heat 12%). Practically all thermosensitive A-fibers and C-fibers were also mechanosensitive. Thus, unlike cutaneous axons, almost all A- and C-fibers afferents in injured muscle nerves demonstrate ectopic activity, even chronically after nerve injury. NEW & NOTEWORTHY After chronic injury of a muscle nerve, allowing the nerve fibers to regenerate to the target tissue, 1) most afferent A-fibers are mechanosensitive and regenerate to the target tissue; 2) ectopic ongoing activity, cold sensitivity, and heat sensitivity significantly decrease with time after injury in A-afferents; 3) most afferent C-fibers do not regenerate to the target tissue; and 4) injured C-afferents maintain the patterns of ectopic discharge properties they already show soon after nerve injury.

Fabrication of Thermosensitive Cyclic Brush Copolymer with Enhanced Therapeutic Efficacy for Anticancer Drug Delivery.

Adaptation of cyclic brush polymer for drug delivery applications remains largely unexplored. Herein, cyclic brush copolymer of poly(2-hydroxyethyl methacrylate-g-poly(N-isopropylacrylamide-st-N-hydroxyethylacrylamide)) (cb-P(HEMA-g-P(NIPAAm-st-HEAAm))), comprising a cyclic core of PHEMA and thermosensitive brushes of statistical copolymer of P(NIPAAm-st-HEAAm), is designed and synthesized successfully via a graft-from approach using atom transfer free radical polymerization from a cyclic multimacroinitiator. The composition of the brush is optimized to endow the resulting cyclic brush copolymer with a lower critical solution temperature (LCST) slightly above the physiological temperature, but lower than the localized temperature of tumor tissue, which is suitable for the hyperthermia-triggered anticancer drug delivery. Critical aggregation concentration determination reveals better stability for the unimolecular nanoparticle formed by the cyclic brush copolymer than that formed by the bottlebrush analogue. The dramatically increased size with elevated temperatures from below to above the LCST confirms hyperthermia-induced aggregation for both formulations. Such structural destabilization promotes significantly the drug release at 40 °C. Most importantly, the drug-loaded cyclic brush copolymer shows enhanced in vitro cytotoxicity against HeLa cells than the bottlebrush counterpart. The better stability and higher therapeutic efficacy demonstrates that the thermosensitive cyclic brush copolymer is a better formulation than bottle brush copolymer for controlled drug release applications.

The Pneumococcal Type 1 Pilus Genes Are Thermoregulated and Are Repressed by a Member of the Snf2 Protein Family.

In Streptococcus pneumoniae, the type 1 pilus is involved in many steps of pathogenesis, including adherence to epithelial cells, mediation of inflammation, escape from macrophages, and the formation of biofilms. The type 1 pilus genes are expressed in a bistable fashion with cells switching between "on" and "off" expression states. Bistable expression of these genes is due to their control by RlrA, a positive regulator subject to control by a positive-feedback loop. The type 1 pilus genes are also thought to be negatively regulated by a large number of repressors. Here we show that expression of the type 1 pilus genes is thermosensitive and switched off at growth temperatures below 31°C. We also report that the on expression state of the type 1 pilus genes is highly stable, a phenomenon which we show likely contributed to the erroneous identification of many proteins as negative regulators of these genes. Finally, we exploited the effect of low temperature on pilus gene expression to help identify SP_1523, an Snf2-type protein, as a novel negative regulator of the pilus genes. Our findings establish that the type 1 pilus genes are thermoregulated and are repressed by a member of the Snf2 protein family. They also refute the notion that these genes are controlled by 8 previously described negative regulators.IMPORTANCEStreptococcus pneumoniae is the leading cause of death from respiratory infections in children. Many bacterial factors contribute to pneumococcal virulence and nasopharyngeal colonization. The type 1 pneumococcal pilus plays an important role in mouse models and in epithelial adherence and is expressed in a bistable fashion. Here we show that the "on" state is highly stable, which may explain the prior misidentification of negative regulators of pilus expression. We also show that expression of pilus genes is thermosensitive: virtually no expression can be detected at temperatures found in the anterior nares of humans. We took advantage of this property to identify a negative regulator of pilus expression, a member of a family of proteins widely conserved across Gram-positive bacteria.

Intravenous Delivery of Xenon Incorporated in Thermosensitive Nano-Emulsions for Anesthesia.

Xenon anesthesia has several advantages over conventional anesthetics, however, it has not been widely used in clinical sites, since the cost and minimum alveolar concentration were higher than conventional inhalational anesthetics. The purpose of this study was to develop an optimum vehicle for stable intravenous delivery of xenon with sufficient concentration. Thermosensitive lipid based nano-emulsions (TS-LE) were prepared by blending medium chain triglyceride, polyethylene glycol-15-hydroxystearate, D-α tocopherol polyethylene glycol succinate and Poloxamer 188. Three folds higher xenon was loaded in TS-LE when it was prepared under 5.5 atm of xenon pressure compared to that under 1 atm of xenon pressure at 22 °C (11.4±0.7 vs. 3.82±0.34 mg/ml). Poloxamer 188 conferred thermosensitive viscosity and outer rigid structure on TS-LE, and the properties led to the enhanced stability of xenon compared to usual lipid emulsion. Induction of anesthesia was investigated by monitoring loss of forepaw righting reflex (LORR) in rats. The ED50 for LORR was 131.9 mg/kg and the anesthesia was maintained for 65.3±7.1 sec at a dose of 179 mg/kg in rats. When it is considered that TS-LE stably loads enough concentration of xenon for the induction of therapeutically sufficient anesthesia, TS-LE may be regarded as a promising candidate for intravenous xenon delivery.

Indirect hand and forearm vasomotion: Regional variations in cutaneous thermosensitivity during normothermia and mild hyperthermia.

In this experiment, hand and forearm vasomotor activity was investigated during localised, but stable heating and cooling of the face, hand and thigh, under open-loop (clamped) conditions. It was hypothesised that facial stimulation would provoke the most potent vascular changes. Nine individuals participated in two normothermic trials (mean body temperature clamp: 36.6°C; water-perfused suit and climate chamber) and two mildly hyperthermic trials (37.9°C). Localised heating (+5°C) and cooling (-5°C) stimuli were applied to equal surface areas of the face, hand and thigh (perfusion patches: 15min), while contralateral forearm or hand blood flows (venous-occlusion plethysmography) were measured (separate trials). Thermal sensation and discomfort votes were recorded before and during each thermal stimulation. When hyperthermic, local heating induced more sensitive vascular responses, with the combined thermosensitivity of both limb segments averaging 0.011mL·100mL-1·min-1·mmHg-1·°C-1, and 0.005mL·100mL-1·min-1·mmHg-1·°C-1 during localised cooling (P<0.05). Inter-site comparisons among the stimulated sites yielded minimal evidence of variations in local thermal sensation, and no differences were observed for vascular conductance (P>0.05). Therefore, regional differences in vasomotor and sensory sensitivity appeared not to exist. When combined with previous observations of sudomotor sensitivity, it seems that, during mild heating and cooling, regional representations within the somatosensory cortex may not translate into meaningful differences in thermal sensation or the central integration of thermoafferent signals. It was concluded that inter-site variations in the cutaneous thermosensitivity of these thermolytic effectors have minimal physiological significance over the ranges investigated thus far.