Fire impacts bacterial composition in Protea repens (Proteaceae) infructescences.

The diverse bacterial communities in and around plants provide important benefits, such as protection against pathogens and cycling of essential minerals through decomposition of moribund plant biomass. Biodiverse fynbos landscapes generally have limited deadwood habitats due to the absence of large trees and frequent fire. In this study, we determined the effect of a fire disturbance on the bacterial communities in a fynbos landscape dominated by the shrub Protea repens using 16S ribosomal RNA amplicon sequencing. The bacterial community composition in newly formed fruiting structures (infructescences) and soil at a recently burnt site was different from that in an unburnt site. Bacteria inhabiting P. repens infructescences were similar to well-known taxa from decomposing wood and litter. This suggests a putative role for these aboveground plant structures as reservoirs for postfire decomposer bacteria. The results imply that inordinately frequent fires, which are commonplace in the Anthropocene, are a significant disturbance to bacterial communities and could affect the diversity of potentially important microbes from these landscapes.

Biodiversity and ecology of flower-associated actinomycetes in different flowering stages of Protea repens.

Actinomycete bacteria have previously been reported from reproductive structures (infructescences) of Protea (sugarbush/suikerbos) species, a niche dominated by fungi in the genera Knoxdaviesia and Sporothrix. It is probable that these taxa have symbiotic interactions, but a lack of knowledge regarding their diversity and general ecology precludes their study. We determined the diversity of actinomycetes within Protea repens inflorescence buds, open inflorescences, young and mature infructescences, and leaf litter surrounding these trees. Since the P. repens habitat is fire-prone, we also considered the potential of these bacteria to recolonise infructescences after fire. Actinomycetes were largely absent from flower buds and inflorescences but were consistently present in young and mature infructescences. Two Streptomyces spp. were the most consistent taxa recovered, one of which was also routinely isolated from leaf litter. Lower colonisation rates were evident in samples from a recently burnt site. One of the most consistent taxa isolated from older trees in the unburnt site was absent from this site. Our findings show that P. repens has a distinct community of actinomycetes dominated by a few species. These communities change over time and infructescence developmental stage, season and the age of the host population. Mature infructescences appear to be important sources of inoculum for some of the actinomycetes, seemingly disrupted by fire. Increased fire frequency limiting maturation of P. repens infructescences could thus impact future actinomycete colonisation in the landscape. Streptomyces spp. are likely to share this niche with the ophiostomatoid fungi, which merits further study regarding their interactions and mode of transfer.

Antifungal Streptomyces spp. Associated with the Infructescences of Protea spp. in South Africa.

Common saprophytic fungi are seldom present in Protea infructescences, which is strange given the abundance of mainly dead plant tissue in this moist protected environment. We hypothesized that the absence of common saprophytic fungi in Protea infructescences could be due to a special symbiosis where the presence of microbes producing antifungal compounds protect the infructescence. Using a culture based survey, employing selective media and in vitro antifungal assays, we isolated antibiotic producing actinomycetes from infructescences of Protea repens and P. neriifolia from two geographically separated areas. Isolates were grouped into three different morphological groups and appeared to be common in the Protea spp. examined in this study. The three groups were supported in 16S rRNA and multi-locus gene trees and were identified as potentially novel Streptomyces spp. All of the groups had antifungal activity in vitro. Streptomyces sp. Group 1 had inhibitory activity against all tested fungi and the active compound produced by this species was identified as fungichromin. Streptomyces spp. Groups 2 and 3 had lower inhibition against all tested fungi, while Group 3 showed limited inhibition against Candida albicans and Sporothrix isolates. The active compound for Group 2 was also identified as fungichromin even though its production level was much lower than Group 1. The antifungal activity of Group 3 was linked to actiphenol. The observed antifungal activity of the isolated actinomycetes could contribute to protection of the plant material against common saprophytic fungi, as fungichromin was also detected in extracts of the infructescence. The results of this study suggest that the antifungal Streptomyces spp. could play an important role in defining the microbial population associated with Protea infructescences.

[Intracranial pressure and hypotonic infusion solutions]. / Intrakranieller Druck und hypotone Infusionslösungen.

The physiological osmolality of plasma is 288+/-5 mosmol/kgH2O when measured by freezing-point depression. The theoretical osmolarity (290 mosmol/l) calculated from composition, osmotic coefficient (0.93) and water content (0.94) is practically identical. Saline (0.9% NaCl) has an osmolarity of 308 mosmol/l and an osmolality of 286 mosmol/kgH2O (water content ca. 1.0). The osmolality in vivo is more important than that measured in vitro. A 5% dextrose solution in water (D5W) is isotonic in vitro, but the in vivo effect is that of pure water because the glucose is rapidly metabolized. Every infusion fluid should be isotonic (290+/-10 mosmol/kgH2O). Hypotonic solutions must move water from the extracellular space to the intracellular space. Typical examples are Ringer's lactate and acetate solutions (256 instead of 290 mosmol/kgH2O). The brain (central nervous system, CNS) is the critical organ: The rigidly shaped skull contains three incompressible compartments, only blood and cerebrospinal fluid (CSF) can be partially, but limitedly shifted outside the skull. The consequence of a volume load is an increasing intracranial pressure (ICP). A decrease in plasma osmolality by only 3% produces an increase in ICP of about 15 mmHg. Therefore, infusion of larger volumes of hypotonic solutions should be avoided at all costs.

Our changing atmosphere: evidence based on long-term infrared solar observations at the Jungfraujoch since 1950.

The Institute of Astrophysics of the University of Liège has been present at the High Altitude Research Station Jungfraujoch, Switzerland, since the late 1940s, to perform spectrometric solar observations under dry and weakly polluted high-mountain conditions. Several solar atlases of photometric quality, extending altogether from the near-ultra-violet to the middle-infrared, were produced between 1956 and 1994, first with grating spectrometers then with Fourier transform instruments. During the early 1970s, scientific concerns emerged about atmospheric composition changes likely to set in as a consequence of the growing usage of nitrogen-containing agricultural fertilisers and the industrial production of chlorine-bearing compounds such as the chlorofluorocarbons and hydro-chlorofluorocarbons. Resulting releases to the atmosphere with ensuing photolysis in the stratosphere and catalytic depletion of the protective ozone layer prompted a worldwide consortium of chemical manufacturing companies to solicit the Liège group to help in clarifying these concerns by undertaking specific observations with its existing Jungfraujoch instrumentation. The following pages evoke the main steps that led from quasi full sun-oriented studies to priority investigations of the Earth's atmosphere, in support of both the Montreal and the Kyoto Protocols.

Observations of long-lived anthropogenic halocarbons at the high-Alpine site of Jungfraujoch (Switzerland) for assessment of trends and European sources.

Anthropogenic halocarbons, such as chlorofluorocarbons (CFCs), hydrochlorofluorocarbons (HCFCs), hydrofluorocarbons (HFCs), bromocarbons (halons) and long-lived chlorinated solvents have been measured continuously at the high-Alpine site of Jungfraujoch (Switzerland) since January 2000. Chloro- and bromo-containing halocarbons are responsible for the stratospheric ozone depletion and will be globally banned from usage within the next years. With the exception of the stable CFC-12 (CF2 Cl2), all major CFCs and chlorinated solvents show a negative trend in recent years in their background concentrations at Jungfraujoch. HCFCs, as their first-generation substitute, are still increasing with a few percent per year. However, the frequency and the strength of HCFCs pollution events, which are caused by regional European emissions, are already declining. This can be seen as a sign of the impending ban of these gases within the next years in Europe. On the other hand, HFCs as the second-generation substitutes, are increasing with relative rates of at least 10% per year (e.g. almost 5 ppt per year for HFC-134a). An allocation of European sources was attempted by combining measured concentrations with trajectories of air masses reaching the Jungfraujoch during pollution events. Potential source regions could be detected in Italy, France, Spain and Germany.

Screening California surface waters for estrogenic endocrine disrupting chemicals (EEDC) with a juvenile rainbow trout liver vitellogenin mRNA procedure.

Concern regarding the occurrence of chemicals that disrupt endocrine system functions in aquatic species has heightened over the last 15 years. However, little attention has been given to monitoring for estrogenic endocrine disrupting chemicals (EEDCs) in California's freshwater ecosystems. The objective was to screen surface water samples for estrogenic activity using vitellogenin (Vtg) mRNA quantification in livers of juvenile rainbow trout by real-time reverse transcriptase polymerase chain reaction (Q-RT PCR). Vtg mRNA analysis of livers from fish exposed to 113 ambient water samples collected from surface waters in California's Central Valley and northern area indicated that six samples (5% of total) may have contained EEDCs. The six samples induced marginal, but statistically significant, increases of Vtg mRNA. No ambient water sample evoked Vtg mRNA responses equivalent to those in positive controls (all responses were less than 2% of the positive control response). Thus, EEDC concentrations in these samples were low (at or near the threshold for the procedure) or results may have included false positives. To establish a more definitive assessment of EEDC occurrence, follow-up screening at sites where statistically significant, but weak, estrogenic activity was observed is recommended. Overall, results reveal that a majority of the California surface waters tested were below EEDC detection threshold concentration for the screening procedure utilized.

[Optimisation of the acid-base status in hypothermia]. / Optimierung des Säure-Basen-Status unter Hypothermie.

Under laboratory conditions, it can be demonstrated within sufficient clinical accuracy that the base excess (BE, [mmol/l]) is independent of temperature. In any blood gas analyzer with temperature correction, the results are consistent with tonometry when measuring a sample of hypothermic equilibrated blood at 37 degrees C. Under clinical conditions, it is shown that there is practically no difference in the CO(2) partial pressure, irrespective of whether measured directly by capnometry (p(et)CO(2)) or obtained from arterial blood (p(a)CO(2)) in the blood gas analyzer after correction for the patient's temperature. Hence, the clinical recommendations for hypothermia are: correct artificial ventilation, preferably pCO(2) 40+/-5 mmHg, should be established by capnometry and controlled by temperature-corrected p(a)CO(2): metabolic changes should be diagnosed by temperature-independent BE: the temperature-corrected pH, if at all, should be used only for the diagnosis of acidosis or alkalosis.

[The design of the VISEP trial. Critical appraisal]. / Studienprotokoll der VISEP-Studie. Eine kritische Stellungnahme.

The survey transcript of the VISEP interventional trial "Prospective randomized multicenter study on the influence of colloid vs crystalloid volume resuscitation and of intensive vs conventional insulin therapy on outcome in patients with severe sepsis and septic shock" [Clinical trials.gov. identifier: NCT00135473; study start April 2003] comprises, according to the data of the year 2003, methodological shortcomings which challenge a priori the study design and thus the resolution of the purpose of the study, i.e., "determination of the influence of the studied volume and insulin interventions on morbidity and mortality of patients with severe sepsis and septic shock". The most important points of criticism are: 1. A volume therapy with exclusively crystalloids or colloids with the chosen colloid hyperoncotic, hyperchloremic HES solution (10% hydroxyethyl starch: 10% Hemohes) or the crystalloid solution with high lactate content (Sterofundin) is neither acceptable nor practicable, even if only due to exceeding the maximum dosage as recommended by the manufacturer. 2. The fact known since the year 2001 that high molecular weight, poorly biodegradable HES preparations can present an independent risk-factor for acute kidney failure in patients with sepsis or septic shock was ignored: the exclusion criterion of a serum-creatinine value of >320 micromol/l (>3.6 mg/dl) was doubled in relation to the manufacturer's specification. 3. The hyperoncotic colloid solution used (10% Hemohes) may only be employed for a brief period: it is highly hyperchloremic and causes extravascular hypohydration with consecutive reduction of renal excretion, which together with HES is a fatal combination. 4. The crystalloid solution used, i.e., Sterofundin, which contains 45 mmol/l lactate, is contraindicated with septic shock as it increases the patient's O2 consumption, hinders lactate diagnostics as a hypoxia marker by simultaneous lactate infusion, and through increased gluconeogenesis leads to hyperglycemia, at least with diabetics. 5. It is doubtful whether an intensified insulin therapy (Actrapid) can be successful if insulin is administered simultaneously with iatrogenic hyperglycemia as a result of lactate influx. Due to these flaws in the design of the VISEP trial, the only consequence can be that the results of the survey are unusable, especially with regard to the point "HES and kidney function". Thus, any further advance presentations and interpretations should be shelved in expectation of the authors' publication of all the data, in order to begin further discussions including the flaws in study design listed here.

[Requirements and expectations for optimal volume replacement]. / Forderungen und Erwartungen an einen optimalen Volumenersatz.

A volume replacement should compensate a reduction in the intravascular volume and counteract a hypovolemia so that hemodynamics and vital functions can be maintained. For this therapy, a physiologically-based solution comprising both osmotically and colloid osmotically active components should be administered. A consensus is proposed for this purpose which takes into consideration the following aspects: The optimum colloid, the questionable use of albumin, the physiological electrolyte pattern encompassing sodium, potassium, chloride and phosphate and their contributions to osmolality, an eventual addition of glucose, the physiological acid-base status with bicarbonate or alternately with metabolisable anions, and the importance of a clear declaration of all ingredients. The consensus distinguishes between compulsory requirements derived from evidence-based medicine and physiological data and the potential expectations of an optimal volume replacement, including well-grounded wishes and aspirations for the future.

[Massive transfusion of washed red blood cells: acid-base and electrolyth changes for different wash solutions]. / Massivtransfusionen mit gewaschenen Erythrozyten: Beeinflussung des Säure-Basen-Elektrolyt-Haushalts durch verschiedene Waschlösungen.

OBJECTIVE: The composition of normal saline (NaCl), the standard wash solution for cell saver autotransfusion, is considerably different from physiologic plasma values in small infants. Therefore, we investigated acid-base and electrolyte changes during massive cell saver autotransfusion with different wash solutions in young pigs. METHODS: After approval by the animal protection authorities 15 young pigs (weight 10.6 +/- 1.1 kg, blood volume 848 +/- 88 ml, mean+/-SD) underwent 15 cycles of cell saver autotransfusion (Haemolite 2plus, Haemonetics). For each cycle, 100 ml arterial blood was withdrawn, washed with NaCl, physiologic multielectrolyte solution (PME, V Infusionslösung 296 mval Elektrolyte, Baxter) or physiologic erythrocyte protection solution (PEP, 3.2 % gelatine, pH 7.40, cHCO3 24 mmol/l), and then retransfused. Analyses of acid-base, electrolyte, and hematologic parameters were performed for systemic and washed blood samples. RESULTS: For NaCl there was a progressive decrease in systemic pH, HCO3 and base excess (BE) and an increase in chloride values (Cl) (p < 0.05). Use of PME slightly decreased pH (n. s.), whereas HCO3, BE and Cl remained stable. PEP slightly increased pH, HCO3 and BE, and decreased Cl (n. s.). Free hemoglobin increased in NaCl and PME (p < 0.05) and was below baseline in PEP (n. s.). Lactic acid course was comparable in all groups. CONCLUSIONS: The use of NaCl as wash solution for massive autotransfusion resulted in metabolic acidosis caused by dilution of HCO3 and increased Cl values. Fewer systemic acid-base and electrolyte changes were observed, when blood was washed with PME or PEP. The decreased hemoglobin release with PEP is possibly due to a gelatine specific electrostatic surface coating of erythrocyte membranes. For massive transfusion of washed red blood cells, physiologic multielectrolyte solution and physiologic erythrocyte protection solution should be preferred to NaCl, especially for small infants.