Decadal soil warming decreased vascular plant above and belowground production in a subarctic grassland by inducing nitrogen limitation.

Below and aboveground vegetation dynamics are crucial in understanding how climate warming may affect terrestrial ecosystem carbon cycling. In contrast to aboveground biomass, the response of belowground biomass to long-term warming has been poorly studied. Here, we characterized the impacts of decadal geothermal warming at two levels (on average +3.3°C and +7.9°C) on below and aboveground plant biomass stocks and production in a subarctic grassland. Soil warming did not change standing root biomass and even decreased fine root production and reduced aboveground biomass and production. Decadal soil warming also did not significantly alter the root-shoot ratio. The linear stepwise regression model suggested that following 10 yr of soil warming, temperature was no longer the direct driver of these responses, but losses of soil N were. Soil N losses, due to warming-induced decreases in organic matter and water retention capacity, were identified as key driver of the decreased above and belowground production. The reduction in fine root production was accompanied by thinner roots with increased specific root area. These results indicate that after a decade of soil warming, plant productivity in the studied subarctic grassland was affected by soil warming mainly by the reduction in soil N.

A systemic overreaction to years versus decades of warming in a subarctic grassland ecosystem.

Temperature governs most biotic processes, yet we know little about how warming affects whole ecosystems. Here we examined the responses of 128 components of a subarctic grassland to either 5-8 or >50 years of soil warming. Warming of >50 years drove the ecosystem to a new steady state possessing a distinct biotic composition and reduced species richness, biomass and soil organic matter. However, the warmed state was preceded by an overreaction to warming, which was related to organism physiology and was evident after 5-8 years. Ignoring this overreaction yielded errors of >100% for 83 variables when predicting their responses to a realistic warming scenario of 1 °C over 50 years, although some, including soil carbon content, remained stable after 5-8 years. This study challenges long-term ecosystem predictions made from short-term observations, and provides a framework for characterization of ecosystem responses to sustained climate change.

Microbial temperature sensitivity and biomass change explain soil carbon loss with warming.

Soil microorganisms control carbon losses from soils to the atmosphere1-3, yet their responses to climate warming are often short-lived and unpredictable4-7. Two mechanisms, microbial acclimation and substrate depletion, have been proposed to explain temporary warming effects on soil microbial activity8-10. However, empirical support for either mechanism is unconvincing. Here we used geothermal temperature gradients (> 50 years of field warming)11 and a short-term experiment to show that microbial activity (gross rates of growth, turnover, respiration and carbon uptake) is intrinsically temperature sensitive and does not acclimate to warming (+ 6 ºC) over weeks or decades. Permanently accelerated microbial activity caused carbon loss from soil. However, soil carbon loss was temporary because substrate depletion reduced microbial biomass and constrained the influence of microbes over the ecosystem. A microbial biogeochemical model12-14 showed that these observations are reproducible through a modest, but permanent, acceleration in microbial physiology. These findings reveal a mechanism by which intrinsic microbial temperature sensitivity and substrate depletion together dictate warming effects on soil carbon loss via their control over microbial biomass. We thus provide a framework for interpreting the links between temperature, microbial activity and soil carbon loss on timescales relevant to Earth's climate system.

Prolonged exposure does not increase soil microbial community compositional response to warming along geothermal gradients.

Global change is expected to affect soil microbial communities through their responsiveness to temperature. It has been proposed that prolonged exposure to elevated temperatures may lead to progressively larger effects on soil microbial community composition. However, due to the relatively short-term nature of most warming experiments, this idea has been challenging to evaluate. The present study took the advantage of natural geothermal gradients (from +1°C to +19°C above ambient) in two subarctic grasslands to test the hypothesis that long-term exposure (>50 years) intensifies the effect of warming on microbial community composition compared to short-term exposure (5-7 years). Community profiles from amplicon sequencing of bacterial and fungal rRNA genes did not support this hypothesis: significant changes relative to ambient were observed only starting from the warming intensity of +9°C in the long term and +7°C/+3°C in the short term, for bacteria and fungi, respectively. Our results suggest that microbial communities in high-latitude grasslands will not undergo lasting shifts in community composition under the warming predicted for the coming 100 years (+2.2°C to +8.3°C).

Complete Genome Sequence of West Nile Virus (WNV) from the First Human Case of Neuroinvasive WNV Infection in Cyprus.

We report here the complete genome sequence of a West Nile virus (WNV) strain from the first laboratory-confirmed human case of neuroinvasive WNV infection in Cyprus. Phylogenetic analysis showed that this WNV strain grouped clearly into genetic lineage 1, clade 1a, cluster 2.

Impact of Soil Warming on the Plant Metabolome of Icelandic Grasslands.

Climate change is stronger at high than at temperate and tropical latitudes. The natural geothermal conditions in southern Iceland provide an opportunity to study the impact of warming on plants, because of the geothermal bedrock channels that induce stable gradients of soil temperature. We studied two valleys, one where such gradients have been present for centuries (long-term treatment), and another where new gradients were created in 2008 after a shallow crustal earthquake (short-term treatment). We studied the impact of soil warming (0 to +15 °C) on the foliar metabolomes of two common plant species of high northern latitudes: Agrostis capillaris, a monocotyledon grass; and Ranunculus acris, a dicotyledonous herb, and evaluated the dependence of shifts in their metabolomes on the length of the warming treatment. The two species responded differently to warming, depending on the length of exposure. The grass metabolome clearly shifted at the site of long-term warming, but the herb metabolome did not. The main up-regulated compounds at the highest temperatures at the long-term site were saccharides and amino acids, both involved in heat-shock metabolic pathways. Moreover, some secondary metabolites, such as phenolic acids and terpenes, associated with a wide array of stresses, were also up-regulated. Most current climatic models predict an increase in annual average temperature between 2-8 °C over land masses in the Arctic towards the end of this century. The metabolomes of A. capillaris and R. acris shifted abruptly and nonlinearly to soil warming >5 °C above the control temperature for the coming decades. These results thus suggest that a slight warming increase may not imply substantial changes in plant function, but if the temperature rises more than 5 °C, warming may end up triggering metabolic pathways associated with heat stress in some plant species currently dominant in this region.

Phenological responses of Icelandic subarctic grasslands to short-term and long-term natural soil warming.

The phenology of vegetation, particularly the length of the growing season (LOS; i.e., the period from greenup to senescence), is highly sensitive to climate change, which could imply potent feedbacks to the climate system, for example, by altering the ecosystem carbon (C) balance. In recent decades, the largest extensions of LOS have been reported at high northern latitudes, but further warming-induced LOS extensions may be constrained by too short photoperiod or unfulfilled chilling requirements. Here, we studied subarctic grasslands, which cover a vast area and contain large C stocks, but for which LOS changes under further warming are highly uncertain. We measured LOS extensions of Icelandic subarctic grasslands along natural geothermal soil warming gradients of different age (short term, where the measurements started after 5 years of warming and long term, i.e., warmed since ≥50 years) using ground-level measurements of normalized difference vegetation index. We found that LOS linearly extended with on average 2.1 days per °C soil warming up to the highest soil warming levels (ca. +10°C) and that LOS had the potential to extend at least 1 month. This indicates that the warming impact on LOS in these subarctic grasslands will likely not saturate in the near future. A similar response to short- and long-term warming indicated a strong physiological control of the phenological response of the subarctic grasslands to warming and suggested that genetic adaptations and community changes were likely of minor importance. We conclude that the warming-driven extension of the LOSs of these subarctic grasslands did not saturate up to +10°C warming, and hence that growing seasons of high-latitude grasslands are likely to continue lengthening with future warming (unless genetic adaptations or species shifts do occur). This persistence of the warming-induced extension of LOS has important implications for the C-sink potential of subarctic grasslands under climate change.

The first human case of neuroinvasive West Nile virus infection identified in Cyprus.

West Nile virus infection can pose a diagnostic challenge to clinicians, especially in geographic areas where human cases of this disease have never been encountered before. In August 2016, the first human case of West Nile virus infection was diagnosed in Cyprus. An elderly non immunosuppressed patient with a history of recent travel, presented with a clinical picture of rapidly progressing ascending paralysis mimicking Guillain-Barré syndrome. Neuroinvasive West Nile virus disease was diagnosed by detecting West Nile virus nucleic acid in the patient's cerebrospinal fluid. Public health measures were taken raising awareness regarding this disease and its prevention. Clinical vigilance to consider West Nile virus as a possible emerging pathogen in the appropriate clinical setting is warranted and could benefit individual patients.

Sensitivity of soil carbon fractions and their specific stabilization mechanisms to extreme soil warming in a subarctic grassland.

Terrestrial carbon cycle feedbacks to global warming are major uncertainties in climate models. For in-depth understanding of changes in soil organic carbon (SOC) after soil warming, long-term responses of SOC stabilization mechanisms such as aggregation, organo-mineral interactions and chemical recalcitrance need to be addressed. This study investigated the effect of 6 years of geothermal soil warming on different SOC fractions in an unmanaged grassland in Iceland. Along an extreme warming gradient of +0 to ~+40 °C, we isolated five fractions of SOC that varied conceptually in turnover rate from active to passive in the following order: particulate organic matter (POM), dissolved organic carbon (DOC), SOC in sand and stable aggregates (SA), SOC in silt and clay (SC-rSOC) and resistant SOC (rSOC). Soil warming of 0.6 °C increased bulk SOC by 22 ± 43% (0-10 cm soil layer) and 27 ± 54% (20-30 cm), while further warming led to exponential SOC depletion of up to 79 ± 14% (0-10 cm) and 74 ± 8% (20-30) in the most warmed plots (~+40 °C). Only the SA fraction was more sensitive than the bulk soil, with 93 ± 6% (0-10 cm) and 86 ± 13% (20-30 cm) SOC losses and the highest relative enrichment in 13 C as an indicator for the degree of decomposition (+1.6 ± 1.5‰ in 0-10 cm and +1.3 ± 0.8‰ in 20-30 cm). The SA fraction mass also declined along the warming gradient, while the SC fraction mass increased. This was explained by deactivation of aggregate-binding mechanisms. There was no difference between the responses of SC-rSOC (slow-cycling) and rSOC (passive) to warming, and 13 C enrichment in rSOC was equal to that in bulk soil. We concluded that the sensitivity of SOC to warming was not a function of age or chemical recalcitrance, but triggered by changes in biophysical stabilization mechanisms, such as aggregation.

Antimicrobial resistance: action to combat the rising microbial challenges.

Antimicrobial therapy transformed medical practice from a merely diagnosis-focused approach 80 years ago to a treatment-focused approach, saving millions of lives in the years to follow. Today, numerous medical advances made possible by effective antibiotics are being threatened by the relentlessly rising rates of bacteria resistant to all currently available antibiotics. This phenomenon is a consequence of antibiotic misuse, which exerts undue selective pressure on micro-organisms, combined with defective infection control practices that accelerate their spread. Its impact on societies worldwide is immense, resulting in loss of human life and money. An alarming pattern of resistance involving multidrug-resistant and sometimes pandrug-resistant Gram-negative bacteria is currently emerging. In response to the global public health threat posed by antimicrobial resistance (AMR), a number of national and international actions and initiatives have been developed in recent years to address this issue. Although the optimally effective and cost-effective strategy to reduce AMR is not known, a multifaceted approach is most likely to be successful. It should include actions aiming at optimising antibiotic use, strengthening surveillance and infection control, and improving healthcare worker and public education with regard to antibiotics. Research efforts to bring new effective antibiotics to patients need to be fostered in order to negate the consequences of the current lack of antimicrobial therapy options. A holistic view of AMR as well as intersectoral collaboration between human and veterinary medicine is required to best address the problem.

Insights into ectopic estrogen receptor expression, nucleocytoplasmic distribution and interaction with chromatin obtained with new antibodies to estrogen receptors α and ß.

Recent reports have indicated that in cells ectopically expressing only ERα or the full-length hormone-binding isoform of ERß (ERß1), the receptors interact with chromatin with different efficacies and that antibodies capable of probing such interactions by chromatin immunoprecipitation (ChIP) are scarce. We therefore produced nine subtype and isoform-specific antibodies to ERα or ERß and validated their performance in receptor probing in cell lines and tissue biopsies by various immunochemical methods, including ChIP. We also produced clones of HEK-293 cells stably transfected with an estrogen response element (ERE)-dependent luciferase reporter and ERα or ERß1, in order to comparatively study their interaction with reporter ERE. We show that ERα was located in the nucleus and ERß1 in the cytoplasm as well as the nucleus of the stably transfected cells, while both receptors were found predominantly in the nucleus in transiently transfected cells and in all estrogen target tissues examined using the same antibodies. The cells displayed wild-type transcriptional activity and canonical regulation of ERE-dependent luciferase expression by estrogen agonists and antagonists. However, unlike ERα, ERß1 recruitment to the reporter ERE could be probed only by sequential ChIP with antibodies to receptor N- and C-terminus. These data suggest that in HEK-293 cells stably expressing ERα or ERß1, ER subtype-specific constraints apply to ERß1 nuclear entry; and that in cells displaying cytoplasmic as well as nuclear localization of ERß1, sequential ChIP with different antibodies to the receptor is the method of choice for probing its interaction with chromatin.

Hydrogen sulphide protects mouse pancreatic ß-cells from cell death induced by oxidative stress, but not by endoplasmic reticulum stress.

BACKGROUND AND PURPOSE: Hydrogen sulphide (H2S), a potentially toxic gas, is also involved in the neuroprotection, neuromodulation, cardioprotection, vasodilatation and the regulation of inflammatory response and insulin secretion. We have recently reported that H2S suppresses pancreatic ß-cell apoptosis induced by long-term exposure to high glucose. Here we examined the protective effects of sodium hydrosulphide (NaHS), an H2S donor, on various types of ß-cell damage. EXPERIMENTAL APPROACH: Isolated islets from mice or the mouse insulinoma MIN6 cells were cultured with palmitate, cytokines (a mixture of tumour necrosis factor-α, interferon-γ and interleukin-1ß), hydrogen peroxide, thapsigargin or tunicamycin with or without NaHS. We examined DNA fragmentation, caspase-3 and -7 activities and reactive oxygen species (ROS) production in the treated cells thereafter. Apoptotic cell death in isolated islets was also assessed by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labelling (TUNEL) method. KEY RESULTS: NaHS suppressed DNA fragmentation and the activities of caspase-3 and -7 induced by palmitate, the cytokines or hydrogen peroxide. In contrast, NaHS failed to protect islets and MIN6 cells from apoptosis induced by thapsigargin and tunicamycin, both of which cause endoplasmic reticulum stress. NaHS suppressed ROS production induced by cytokines or hydrogen peroxide but it had no effect on ROS production in thapsigargin-treated cells. NaHS increased Akt phosphorylation in MIN6 cells treated with cytokines but not in cells treated with thapsigargin. Treatment with NaHS decreased TUNEL-positive cells in cytokine-exposed islets. CONCLUSIONS AND IMPLICATIONS: H2S may prevent pancreatic ß-cells from cell apoptosis via an anti-oxidative mechanism and the activation of Akt signalling.

Rules for identifying patients at increased risk for candidal infections in the surgical intensive care unit: approach to developing practical criteria for systematic use in antifungal prophylaxis trials.

The high rates of invasive candidiasis among intensive care unit (ICU) patients suggest that antifungal prophylaxis might be of value, but rules identifying patients who would best benefit are not established. Based on a retrospective study of 327 patients who stayed in a surgical ICU for > or = 4 days and had an 11.0% rate of invasive candidiasis, we sought to identify useful predictive rules. As prior work suggests that prompt initiation of prophylaxis is of value, we required our rules to be based on patient data routinely available during the week prior to ICU admission through the third day of the ICU stay. Patients with any combination of diabetes mellitus, new onset hemodialysis, use of total parenteral nutrition, or receipt of broad-spectrum antibiotics had an invasive candidiasis rate of 16.6% versus a 5.1% rate for patients lacking these characteristics (P = 0.001). Fifty-two percent of patients staying > or = 4 days in the ICU met this rule and the rule captured 78% of the patients who eventually developed invasive candidiasis. Risk-stratified antifungal prophylaxis in the ICU is possible. Validation of these results in other types of ICU is now needed.

Peritonitis due to Aspergillus and zygomycetes in patients undergoing peritoneal dialysis: report of 2 cases and review of the literature.

Fungal peritonitis in patients on continuous ambulatory peritoneal dialysis (CAPD) has been associated with high mortality and high CAPD-discontinuation rates. Most cases are due to Candida spp. while Aspergillus spp. and zygomycetes have only rarely been implicated. We report one case each of CAPD-related peritonitis caused by Aspergillus terreus and Mucor sp., which have previously been described in the literature once and twice, respectively. The former had a slowly progressive course, did not respond to amphotericin B (AB), and died; the latter improved after a prolonged course of liposomal-AB. Among reported cases of CAPD-related peritonitis due to molds (22 Aspergillus spp. and seven zygomycetes), previous bacterial peritonitis was a common event, the related mortality associated with Aspergillus and zygomycetes was 27% and 57%, respectively, prompt removal of the Tenckoff catheter was critical for survival, and most patients were not able to resume CAPD.

In vitro activities of investigational triazoles against Fusarium species: effects of inoculum size and incubation time on broth microdilution susceptibility test results.

We studied the effects of inoculum size and incubation time on the susceptibility testing results for various antifungal agents against 22 Fusarium isolates by the NCCLS microdilution method. Increased inoculum size and extended incubation time resulted in elevated MICs. Posaconazole and voriconazole exhibited promising antifungal activities.

Solitary parietal lobe histoplasmoma mimicking a brain tumor.

We describe a rare case of Histoplasma infection, which manifested only as a brain histoplasmoma, in a previously healthy woman who had no underlying predisposing conditions. Only a few cases of such intracranial histoplasmomas have been reported but this entity can mimic a brain neoplasm and should be a diagnostic consideration during the evaluation of ring-enhancing brain lesions.

In vitro antifungal susceptibilities of Trichosporon species.

The in vitro activities of amphotericin B, itraconazole, fluconazole, voriconazole, posaconazole, and ravuconazole against 39 isolates of Trichosporon spp. were determined by the NCCLS M27-A microdilution method. The azoles tested appeared to be more potent than amphotericin B. Low minimal fungicidal concentration/MIC ratios were observed for voriconazole, posaconazole, and ravuconazole, suggesting fungicidal activity.

Characterization of secretory and morphologic properties of primary cultured endocrine cells from porcine pancreata.

We characterized morphologic and secretory properties of porcine pancreatic endocrine cells in primary culture obtained by autolytic preparation without any exogenous proteolytic enzymes. The endocrine cells exhibited a neuron-like shape, and insulin granules were accumulated at the terminal of the processes. Thus derived endocrine cells survived in culture medium containing nicotinamide and remained sensitive to glucose for at least 6 weeks after preparations. The cells responded well to physiologic concentrations of glucose, and high K+ depolarization and the antidiabetic sulfonylureas, tolbutamide, and glibenclamide also elicited the release. With high glucose, insulin release was markedly potentiated by forskolin, glucagon, glucagon-like peptide-1, and arginine and inhibited by somatostatin, the Ca2+ channel blocker nitrendipine, and the ATP-sensitive K+ channel opener diazoxide. Epinephrine had dual effects on the release by glucose; enhanced within a low nanomolar range and inhibited at 1 micromol/L. However, the cells were unresponsive to leucine. Such secretory sensitivities to nutrients, hormones, and pharmacologic agents, and long survival rate (as long as 5-6 weeks) of these cells suggest to us therefore that derived endocrine cells may be useful for xenotransplantation of pancreatic beta cells for treatment of insulin-dependent diabetes mellitus.

Lack of effect of incretin hormones on insulin release from pancreatic islets in the bile duct-ligated rats.

Hyperglycemia associated with obstructive jaundice seriously affects the prognosis of patients with hepatobiliary diseases. We investigated secretory properties of isolated islets from bile duct-ligated (BDL) rats. Pancreatic islets from BDL rats lost their secretory responses to glucagon-like peptide-1 (GLP-1), although their responses to glucose were normal. Loss of potentiation of insulin release was also observed in glucagon and glucose-dependent insulinotropic peptide (GIP), whereas modulation of the release by forskolin, dibutyryl cAMP, or epinephrine remained unaffected. cAMP production by BDL islets was not increased by these insulinotropic hormones. Serum levels of glucagon, but not GIP, were increased in BDL rats. GLP-1 levels were also elevated, although they did not reach statistical significance. Immunoblotting of trimeric G protein subunits demonstrated that G(s)alpha L and G(s)alpha S, but not G(i)alpha 1/2 and G(i)alpha 3/o alpha, were less expressed in BDL islets. Therefore, unresponsiveness of the beta-cell to cAMP-raising hormones is involved in glucose intolerance under cholestasis. It results from diminished expression of alpha-subunits of the relevant G protein, G(s), and desensitization of receptors of these hormones.

[Regulation of insulin release: analysis of the insulin secretory cascade and possible contribution to novel anti-diabetic drug development].

Insulin is the only 'hypoglycemic' hormone synthesized in and secreted from the pancreatic beta cell. Type 2 diabetes results from both secretory failure in the beta cell and insulin resistance in the target tissues for insulin. Attempts to develop anti-diabetic drugs that induce insulin secretion from residual beta cells in type 2 diabetic patients originate from the serendipitous discovery of sulphonylureas as hypoglycemic agents 60 years ago. Generally, secretion is carried out by sequential processes such as granule formation, intracellular traffic, granule docking/priming and the final step, exocytosis (secretory cascade). In the beta cell, recent progress in cell biology enables us to analyze each step in the secretory cascade and to reveal controlling mechanisms. This review describes regulatory mechanisms of insulin release by distinct nutrients, hormones and neurotransmitters, and roles of second messengers and protein phosphorylation in the insulin secretory cascade. Possible development of insulinotropic drugs for the treatment of type 2 diabetes has been also discussed.