RESUMEN
BACKGROUND: Dexamethasone is commonly given as an antiemetic during surgical procedures. It has immunosuppressive effects and can affect key enzymes involved in the synthesis of specialised lipid mediators of inflammation resolution (SPM) that direct inflammation resolution and have anti-nociceptive actions. This study examined the effect of dexamethasone on plasma SPM, and the relationship between SPM and perceived pain in women undergoing surgery. METHODS: Plasma SPM were measured in samples obtained from two double-blind controlled interventions. The first, included 51 women mean age 53 ± 1.5 years, undergoing breast surgery allocated to either intravenous saline, or dexamethasone (4 mg or 8 mg) after induction of anaesthesia. The second study included 31 women of mean age 44 ± 0.5 years undergoing laparoscopic gynecological surgery that were allocated to either saline, or dexamethasone (4 mg). SPM (18-HEPE, 17-HDHA, RvE2, RvD1 17R-RvD1 and RvD2) were measured in plasma collected prior to induction of anaesthesia and at 24 h, and 6 weeks post-surgery. Pain was assessed using a verbal analogue scale at discharge from the post-anaesthesia recovery unit. The data from each study was combined to examine the effect of dexamethasone on plasma SPM. The relationship between pain score and SPM was examined using ordinal logistic regression. RESULTS: The SPM 18-HEPE, 17-HDHA, RvE2, RvD1 17R-RvD1 and RvD2 were detectable in all plasma samples. There was no significant difference in any SPM due to dexamethasone over the duration of the study. There was a fall in 17-HDHA between baseline and 24 h in both the dexamethasone and saline groups (P = 0.003) but no change in the downstream SPM (RvD1, 17R-RvD1 and RvD2) or 18-HEPE and RvE2. Pain score was negatively related to levels of RvE2 measured prior to induction of anaesthesia (rho = -0.2991, P = 0.006) and positively related to BMI (rho = 0.279, P = 0.011). In ordinal logistic regression the odds ratio for RvE2 was 0.931 (CI 0.880, 0.986; P = 0.014); after adjusting for the effect of BMI indicating that an increase in RvE2 of 1 pg/ml would result in a 6.9 % fall in pain score. Allocation to a dexamethasone group did not influence the pain score or the relationship between RvE2 and pain score. CONCLUSION: Dexamethasone administered as an anti-emetic does not affect plasma SPM levels. An elevated RvE2 level prior to surgery is predictive of a lower perceived pain score post-anaesthesia.
Asunto(s)
Antieméticos/farmacología , Dexametasona/farmacología , Mediadores de Inflamación/sangre , Dolor Postoperatorio/tratamiento farmacológico , Adulto , Antieméticos/uso terapéutico , Dexametasona/uso terapéutico , Relación Dosis-Respuesta a Droga , Método Doble Ciego , Femenino , Humanos , Persona de Mediana EdadRESUMEN
This study explores the temperature dependency of the aging rate in dry cells over a broad temperature range encompassing the fluid to solid transition (Tg) and well below. Spores from diverse species of eight families of ferns were stored at temperatures ranging from +45�C to approximately -176�C (vapor phase above liquid nitrogen), and viability was monitored periodically for up to 4,300 d (â¼12 years). Accompanying measurements using differential scanning calorimetry (DSC) provide insights into structural changes that occur, such as Tg between +45 and -20�C (depending on moisture), and triacylglycerol (TAG) crystallization between -5 and -35�C (depending on species). We detected aging even at cryogenic temperatures, which we consider analogous to unscheduled degradation of pharmaceuticals stored well below Tg caused by a shift in the nature of molecular motions that dominate chemical reactivity. We occasionally observed faster aging of spores stored at -18�C (conventional freezer) compared with 5�C (refrigerator), and linked this with mobility and crystallization within TAGs, which probably influences molecular motion of dried cytoplasm in a narrow temperature range. Temperature dependency of longevity was remarkably similar among diverse fern spores, despite widely disparate aging rates; this provides a powerful tool to predict deterioration of germplasm preserved in the solid state. Future work will increase our understanding of molecular organization and composition contributing to differences in longevity.
Asunto(s)
Helechos/fisiología , Esporas/fisiología , Rastreo Diferencial de Calorimetría , Polystichum/fisiología , Pteris/fisiología , Temperatura , Factores de TiempoRESUMEN
BACKGROUND AND AIMS: Determining seed longevity by identifying chemical changes that precede, and may be linked to, seed mortality, is an important but difficult task. The standard assessment, germination proportion, reveals seed longevity by showing that germination proportion declines, but cannot be used to predict when germination will be significantly compromised. Assessment of molecular integrity, such as RNA integrity, may be more informative about changes in seed health that precede viability loss, and has been shown to be useful in soybean. METHODS: A collection of seeds stored at 5 °C and 35-50 % relative humidity for 1-30 years was used to test how germination proportion and RNA integrity are affected by storage time. Similarly, a collection of seeds stored at temperatures from -12 to +32 °C for 59 years was used to manipulate ageing rate. RNA integrity was calculated using total RNA extracted from one to five seeds per sample, analysed on an Agilent Bioanalyzer. RESULTS: Decreased RNA integrity was usually observed before viability loss. Correlation of RNA integrity with storage time or storage temperature was negative and significant for most species tested. Exceptions were watermelon, for which germination proportion and storage time were poorly correlated, and tomato, which showed electropherogram anomalies that affected RNA integrity number calculation. Temperature dependencies of ageing reactions were not significantly different across species or mode of detection. The overall correlation between germination proportion and RNA integrity, across all experiments, was positive and significant. CONCLUSIONS: Changes in RNA integrity when ageing is asymptomatic can be used to predict onset of viability decline. RNA integrity appears to be a metric of seed ageing that is broadly applicable across species. Time and molecular mobility of the substrate affect both the progress of seed ageing and loss of RNA integrity.
Asunto(s)
Germinación , Semillas , Cinética , Estabilidad del ARN , Glycine max , TemperaturaRESUMEN
The nature and kinetics of reactions in dry seeds determines how long the seeds survive. We used gas chromatography to assay volatile organic compounds (VOCs) emitted from seeds of three unrelated species as a means to non-invasively probe chemical changes during very dry, dry, and humid storage (seeds were dried to 5.5, 33, and 75% relative humidity at room temperature). VOCs emitted from seeds stored in humid conditions reflected fermentation-type reactions, with methanol and ethanol being predominant in Lactuca sativa and Carum carvi, and acetaldehyde and acetone being predominant in Eruca vesicaria. Dried C. carvi seeds continued to emit fermentation-type products, although at slower rates than the seeds stored in humid conditions. In contrast, drying caused a switch in VOC emission in L. sativa and E. vesicaria seeds towards higher emission of pentane and hexanal, molecules considered to be byproducts from the peroxidation of polyunsaturated fatty acids. Longevity correlated best with the rate of fermentation-type reactions and appeared unrelated to the rate of lipid peroxidation. Emission of VOCs decreased when seed species were mixed together, indicating that seeds adsorbed VOCs. Adsorption of VOCs did not appear to damage seeds, as longevity was not affected in seed mixtures. Collectively, the study shows similarity among species in the types of reactions that occur in dry seeds, but high diversity in the substrates, and hence the byproducts, of the reactions. Moreover, the study suggests that the most abundant VOCs arise from degradation of storage reserves within seed cells, and that these reactions and their byproducts are not, in themselves, damaging.
Asunto(s)
Desecación , Semillas/metabolismo , Compuestos Orgánicos Volátiles/metabolismo , Carum/metabolismo , Ácidos Grasos/metabolismo , Germinación , Humedad , Cinética , Lactuca/metabolismo , Temperatura , Factores de TiempoRESUMEN
BACKGROUND AND AIMS: Quercus species are often considered 'foundation' components of several temperate and/or subtropical forest ecosystems. However, the populations of some species are declining and there is considerable urgency to develop ex situ conservation strategies. In this study, the storage physiology of seeds within Quercus was explored in order to determine factors that affect survival during cryopreservation and to provide a quantitative assessment of seed recalcitrance to support future studies of this complex trait. METHODS: Water relations and survival of excised axes in response to water loss and cryo-exposure were compared for four Quercus species from subtropical China (Q. franchetii, Q. schottkyana) and temperate USA (Q. gambelii, Q. rubra). KEY RESULTS: Seed tissues initially had high water contents and water potentials. Desiccation tolerance of the embryonic axis was not correlated with the post-shedding rainfall patterns where the samples originated. Instead, higher desiccation tolerance was observed in samples growing in areas with colder winters. Survival following cryo-exposure correlated with desiccation tolerance. Among species, plumule tissues were more sensitive than radicles to excision, desiccation and cryo-exposure, and this led to a higher proportion of abnormally developing embryos during recovery following stress. CONCLUSIONS: Quercus species adapted to arid and semi-humid climates still produce recalcitrant seeds. The ability to avoid freezing rather than drought may be a more important selection factor to increase desiccation tolerance. Cryopreservation of recalcitrant germplasm from temperate species is currently feasible, whilst additional protective treatments are needed for ex situ conservation of Quercus from tropical and subtropical areas.
Asunto(s)
Adaptación Fisiológica , Quercus/embriología , Quercus/fisiología , Semillas/fisiología , Estrés Fisiológico , Biomasa , China , Clima , Deshidratación , Electrólitos/metabolismo , Congelación , Raíces de Plantas/fisiología , Presión , Quercus/crecimiento & desarrollo , Análisis de Regresión , Factores de Tiempo , Estados Unidos , AguaRESUMEN
The duration that seeds stay vigorous during storage is difficult to predict but critical to seed industry and conservation communities. Production of volatile compounds from lettuce seeds during storage was investigated as a non-invasive and early detection method of seed ageing rates. Over 30 volatile compounds were detected from lettuce seeds during storage at 35 degrees C at water contents ranging from 0.03 to 0.09 g H(2)O g(-1) dw. Both qualitative and quantitative differences in volatile composition were noted as a function of water content, and these differences were apparent before signs of deterioration were visible. Seeds stored at high water content (L >or=0.06 g H(2)O g(-1) dw) emitted molecular species indicative of glycolysis (methanol+ethanol), and evidence of peroxidation was apparent subsequent to viability loss. Seeds containing less water (0.03-0.05 g H(2)O g(-1) dw) produced volatiles indicative of peroxidation and survived longer compared with seeds stored under more humid conditions. Production of glycolysis-related by-products correlated strongly with deterioration rate when measured as a function of water content. This correlation may provide a valuable non-invasive means to predict the duration of the early, asymptomatic stage of seed deterioration.
Asunto(s)
Lactuca/embriología , Semillas/metabolismo , Compuestos Orgánicos Volátiles/metabolismo , Lactuca/química , Lactuca/metabolismo , Peroxidación de Lípido , Semillas/química , Semillas/crecimiento & desarrollo , Temperatura , Compuestos Orgánicos Volátiles/análisis , AguaRESUMEN
INTRODUCTION: The synthetic glucocorticoid dexamethasone is a commonly administered antiemetic. It has immunosuppressive effects and may alter postoperative blood glucose concentrations. Dexamethasone can effect key enzymes involved in inflammation resolution that is an active process driven by specialised lipid mediators of inflammation resolution (SPM). The purpose of this study in healthy volunteers was to examine whether dexamethasone effects cell populations and synthesis of SPM that are critical for the resolution of inflammation. METHODS: Thirty-two healthy volunteers were randomly allocated to receive saline (Control) or dexamethasone 2â¯mg, 4â¯mg or 8â¯mg intravenously. Venous blood samples were collected at baseline before administration of treatment, and at 4 h, 24 h and one-week post-treatment. At each time point, measurements included blood glucose and macrophage migration inhibition factor (MMIF), full blood count including lymphocyte subsets, monocytes, neutrophils, eosinophils and basophils by flow cytometry, and plasma SPM using liquid chromatography tandem mass spectrometry. The effect of dexamethasone dose and time on all measures was analysed using linear mixed models. RESULTS: There was a dose-dependent increase in neutrophil count after dexamethasone that persisted for 24 h. In contrast, there was a dose-dependent reduction in counts of monocytes, lymphocytes, basophils and eosinophils 4 h after dexamethasone, followed by a rebound increase in cell counts at 24 h. Seven days after administration of dexamethasone, all cell counts were similar to baseline levels. MMIF concentration, glucose and natural killer cell counts were not significantly affected by dexamethasone. There was a significant gender effect on plasma SPM such that levels of 17-HDHA, RvD1, 17R-RvD1 and RvE2 in females were on average 14%-50% lower than males. In a linear mixed model that adjusted for neutrophil count, there was a significant interaction between the dose of dexamethasone and time, on plasma 17R-RvD1 such that plasma 17R-RvD1 fell in a dose-dependent manner until 4 h after administration of dexamethasone. There were no significant effects of dexamethasone on the other plasma SPM (18-HEPE, RvE2, 17-HDHA, RvD1, RvD2 and 14-HDHA) measured. DISCUSSION: This is the first study in healthy volunteers to demonstrate that commonly employed antiemetic doses of dexamethasone affect immune cell populations and plasma levels of 17R-RvD1 an SPM with anti-nociceptive properties. If similar changes occur in surgical patients, then this may have implications for acute infection risk in the post-operative period.
Asunto(s)
Antieméticos , Glucemia , Dexametasona , Ácidos Grasos Insaturados , Mediadores de Inflamación , Leucocitos , Adulto , Antieméticos/administración & dosificación , Antieméticos/efectos adversos , Glucemia/inmunología , Glucemia/metabolismo , Dexametasona/administración & dosificación , Dexametasona/efectos adversos , Ácidos Grasos Insaturados/sangre , Ácidos Grasos Insaturados/inmunología , Femenino , Humanos , Mediadores de Inflamación/sangre , Mediadores de Inflamación/inmunología , Oxidorreductasas Intramoleculares/sangre , Oxidorreductasas Intramoleculares/inmunología , Recuento de Leucocitos , Leucocitos/inmunología , Leucocitos/metabolismo , Factores Inhibidores de la Migración de Macrófagos/sangre , Factores Inhibidores de la Migración de Macrófagos/inmunología , Masculino , Persona de Mediana EdadRESUMEN
This work contributes to the understanding of plant cell responses to extreme water stress when it is applied at different intensity and duration. Fern spores are used to explore survival at relative humidity (RH)<85% because their unicellular nature eliminates complexities that may arise in multicellular organisms from slower drying and variable responses of different cell types. Fern spore cytoplasm solidifies between 30 and 60% RH and spores survive this transition, but subsequently lose viability. We characterized the kinetics of viability loss in terms of the fluid to solid transition using concepts of water activity (i.e., sorption) and glass transition (Tg), two concepts that dominate studies of food and pharmaceutical stability. For all fern species studied, longest survival times were observed in spores placed at about 10-25% RH and mortality rates increased sharply above and below this moisture level. A RH of 10-25% corresponds well to sorption behavior parameters and is below the glass transition, measured using differential scanning calorimetry. Though response to RH was similar among species, the kinetics of deterioration varied considerably among species and this implies differences in the structure or mobility of molecules within the solidified cytoplasm. Our work suggests that desiccation damage occurs in desiccation tolerant cells, and that it is expressed as a time-dependent response, otherwise known as aging.
Asunto(s)
Adaptación Fisiológica , Desecación , Helechos/fisiología , Esporas/fisiología , Rastreo Diferencial de Calorimetría , Ecosistema , Germinación , Humedad , Especificidad de la Especie , Temperatura , Agua/metabolismoRESUMEN
The transition from anhydrobiotic to hydrated state occurs during early imbibition of seeds and is lethal if lipid reserves in seeds are crystalline. Low temperatures crystallize lipids during seed storage. We examine the nature of cellular damage observed in seeds of Cuphea wrightii and C. lanceolata that differ in triacylglycerol composition and phase behavior. Intracellular structure, observed using transmission electron microscopy, is profoundly and irreversibly perturbed if seeds with crystalline triacylglycerols are imbibed briefly. A brief heat treatment that melts triacylglycerols before imbibition prevents the loss of cell integrity; however, residual effects of cold treatments in C. wrightii cells are reflected by the apparent coalescence of protein and oil bodies. The timing and temperature dependence of cellular changes suggest that damage arises via a physical mechanism, perhaps as a result of shifts in hydrophobic and hydrophilic interactions when triacylglycerols undergo phase changes. Stabilizers of oil body structure such as oleosins that rely on a balance of physical forces may become ineffective when triacylglycerols crystallize. Recent observations linking poor oil body stability and poor seed storage behavior are potentially explained by the phase behavior of the storage lipids. These findings directly impact the feasibility of preserving genetic resources from some tropical and subtropical species.
Asunto(s)
Cuphea/metabolismo , Semillas/metabolismo , Triglicéridos/metabolismo , Cristalización , Cuphea/citología , Cuphea/embriología , Cuphea/ultraestructura , TemperaturaRESUMEN
Persistence of anhydrous organisms in nature may depend on how long they remain viable in dry environments. Longevity is determined by interactions of humidity, temperature, and unknown cellular factors that affect the propensity for damaging reactions. Here we describe our research to elucidate those cellular factors and to ultimately predict how long a population can survive under extreme conditions. Loss of viability typically follows a sigmoidal pattern, where a period of small changes precedes a cataclysmic decline. The time for viability to decrease to 50% (P50) varied among seed species and among 10 phylogenetically diverse organisms. When stored at elevated temperatures of 35°C and 32% relative humidity (RH), P50 ranged from about a week for spores of Serratia marcescens to several years for fronds of Selaginella lepidophylla. Most of the species studied survived longest at low humidity (10-20% RH), but suffered under complete dryness. Temperature dependencies of aging kinetics appeared similar among diverse organisms despite the disparate longevities. The effect of temperature on seed aging rates was consistent with the temperature dependency of molecular mobility of aqueous glasses, with both showing a reduction by several orders of magnitude when seeds were cooled from 60°C to 0°C. Longevity is an inherited trait in seeds, but its complex expression among widely divergent taxa suggests that it developed through multiple pathways.
RESUMEN
The purpose of this study was to examine the effect of a 5-day creatine (CR) supplementation period on red blood cell (RBC) CR uptake in vegetarian and nonvegetarian young women. Blood samples were collected from lacto-ovo vegetarians (VG, n = 6, age 21.8 +/- 1.9 yrs) and nonvegetarians (NV, n = 6, age 21.7 +/- 1.9 yrs) before and after a 5-day CR loading period (0. 3g CR/kg lean body mass/day), and from a control group of nonvegetarians (NV, n = 5, age 22.0 +/- 0.7 yrs) who did not supplement with creatine. RBC and plasma samples were analyzed for the presence of creatine. Significant increases (p < .05) in RBC and plasma CR levels were found for vegetarians and nonvegetarians following supplementation. The initial RBC CR content was significantly lower (p < .05) in the vegetarian group. There was no significant difference between vegetarians and nonvegetarians in final RBC CR content, suggesting that a ceiling had been reached. As the uptake into both muscle and RBC is moderated by creatine transporter proteins, analysis of the uptake of CR into RBC may reflect the uptake of CR into muscle, offering an alternative to biopsies.