Randomized trial of red cell washing for the prevention of transfusion-associated organ injury in cardiac surgery.

BACKGROUND: Experimental studies suggest that mechanical cell washing to remove pro-inflammatory components that accumulate in the supernatant of stored donor red blood cells (RBCs) might reduce inflammation and organ injury in transfused patients. METHODS: Cardiac surgery patients at increased risk of large-volume RBC transfusion were eligible. Participants were randomized to receive either mechanically washed allogenic RBCs or standard care RBCs. The primary outcome was serum interleukin-8 measured at baseline and at four postsurgery time points. A mechanism substudy evaluated the effects of washing on stored RBCs in vitro and on markers of platelet, leucocyte, and endothelial activation in trial subjects. RESULTS: Sixty adult cardiac surgery patients at three UK cardiac centres were enrolled between September 2013 and March 2015. Subjects received a median of 3.5 (interquartile range 2-5.5) RBC units, stored for a mean of 21 ( sd 5.2) days, within 48 h of surgery. Mechanical washing reduced concentrations of RBC-derived microvesicles but increased cell-free haemoglobin concentrations in RBC supernatant relative to standard care RBC supernatant. There was no difference between groups with respect to perioperative serum interleukin-8 values [adjusted mean difference 0.239 (95% confidence intervals -0.231, 0.709), P =0.318] or concentrations of plasma RBC microvesicles, platelet and leucocyte activation, plasma cell-free haemoglobin, endothelial activation, or biomarkers of heart, lung, or kidney injury. CONCLUSIONS: These results do not support a hypothesis that allogenic red blood cell washing has clinical benefits in cardiac surgery. CLINICAL TRIAL REGISTRATION: ISRCTN 27076315.

Bacterial diversity and antibiotic resistances of abundant aerobic culturable bacteria in input and output samples of 15 German biogas plants.

AIM: Determination of the diversities and antibiotic resistances of the most abundant aerobic culturable bacteria and their survival in 15 German biogas plants (BGPs). METHODS AND RESULTS: Microbiological standard media (complex and selective media) were applied to enumerate mesophilic and oligotrophic bacteria (Enterobacteriaceae, Salmonella, staphylococci and enterococci) in input and output samples of BGPs. Concentrations of aerobic culturable bacteria in outputs were mostly one to two orders of magnitude lower than in input samples. In total, 852 isolates from input and 902 from output samples were identified either by partial 16S rRNA gene sequencing or Matrix-assisted Laser Desorption/Ionization-Time of Flight (MALDI-TOF) analysis. Antibiotic resistance screening of target families against veterinary relevant antibiotics resulted in similar resistance pattern in input and output samples. CONCLUSIONS: Potentially pathogenic bacteria were detected in output samples of BGPs, but in decreased abundance. The selectivity of the most applied clinical standard media was insufficient. SIGNIFICANCE AND IMPACT OF THE STUDY: Only little is known about the release of antibiotic resistant and potentially pathogenic bacteria from BGPs. However, this study indicates the detection of several potentially pathogenic bacteria in output samples, which showed several antibiotic resistances.

Trial protocol for a randomised controlled trial of red cell washing for the attenuation of transfusion-associated organ injury in cardiac surgery: the REDWASH trial.

INTRODUCTION: It has been suggested that removal of proinflammatory substances that accumulate in stored donor red cells by mechanical cell washing may attenuate inflammation and organ injury in transfused cardiac surgery patients. This trial will test the hypotheses that the severity of the postoperative inflammatory response will be less and postoperative recovery faster if patients undergoing cardiac surgery receive washed red cells compared with standard care (unwashed red cells). METHODS AND ANALYSIS: Adult (≥16 years) cardiac surgery patients identified at being at increased risk for receiving large volume red cell transfusions at 1 of 3 UK cardiac centres will be randomly allocated in a 1:1 ratio to either red cell washing or standard care. The primary outcome is serum interleukin-8 measured at 5 postsurgery time points up to 96 h. Secondary outcomes will include measures of inflammation, organ injury and volumes of blood transfused and cost-effectiveness. Allocation concealment, internet-based randomisation stratified by operation type and recruiting centre, and blinding of outcome assessors will reduce the risk of bias. The trial will test the superiority of red cell washing versus standard care. A sample size of 170 patients was chosen in order to detect a small-to-moderate target difference, with 80% power and 5% significance (2-tailed). ETHICS AND DISSEMINATION: The trial protocol was approved by a UK ethics committee (reference 12/EM/0475). The trial findings will be disseminated in scientific journals and meetings. TRIAL REGISTRATION NUMBER: ISRCTN 27076315.

Rhodococcus defluvii sp. nov., isolated from wastewater of a bioreactor and formal proposal to reclassify [Corynebacterium hoagii] and Rhodococcus equi as Rhodococcus hoagii comb. nov.

A Gram-stain-positive, non-endospore-forming rod-shaped bacterium, strain Ca11(T), was isolated from a bioreactor with extensive phosphorus removal and was studied in detail for its taxonomic allocation. 16S rRNA gene sequence analysis revealed closest sequence similarity of the strain to type strains of [Corynebacterium hoagii] and Rhodococcus equi (98.9%), Rhodococcus koreensis and Rhodococcus wratislaviensis (both 98.4%), Rhodococcus opacus and Rhodococcus canchipurensis (both 98.0%) followed by Rhodococcus kunmingensis and Rhodococcus imtechensis (97.7%). Phylogenetic trees showed a distinct clustering of strain Ca11(T) with the type strains of [C. hoagii], R. equi, and R. kunmingensis separate to all other species of the genus Rhodococcus. The quinone system of strain Ca11(T) was composed of dihydrogenated menaquinones with 8 (major amount) as well as 7 and 6 isoprenoid units [MK-8(H2), MK-7(H2), MK-6(H2)]. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside, one unknown phospholipid and an unidentified glycolipid. The fatty acid profile was similar to that reported for R. equi and contained major amounts of C16:0, C18:1ω9c and 10-methyl C18:0, supporting the allocation of the strain to the genus Rhodococcus. Physiological and biochemical characterization and DNA-DNA hybridization with type strains of the most closely related species allowed clear phenotypic and genotypic differentiation of the isolate. On the basis of these results, strain Ca11(T) ( = DSM 45893(T) = LMG 27563(T)) represents a novel species of the genus Rhodococcus, with the proposed name Rhodococcus defluvii sp. nov. In addition, a polyphasic taxonomic analysis of [Corynebacterium hoagii] DSM 20295(T) and Rhodococcus equi DSM 20307(T) indicated that the two strains belong to the same species, for which the name Rhodococcus hoagii comb. nov. takes priority, according to the Rules of the Bacteriological Code.

Pro-inflammatory effects and oxidative stress in lung macrophages and epithelial cells induced by ambient particulate matter.

The objective of this study was to compare the toxicological effects of different source-related ambient PM10 samples in regard to their chemical composition. In this context we investigated airborne PM from different sites in Aachen, Germany. For the toxicological investigation human alveolar epithelial cells (A549) and murine macrophages (RAW264.7) were exposed from 0 to 96 h to increasing PM concentrations (0-100 µg/ml) followed by analyses of cell viability, pro-inflammatory and oxidative stress responses. The chemical analysis of these particles indicated the presence of 21 elements, water-soluble ions and PAHs. The toxicological investigations of the PM10 samples demonstrated a concentration- and time-dependent decrease in cell viability and an increase in pro-inflammatory and oxidative stress markers.

Gordonia phosphorivorans sp. nov., isolated from a wastewater bioreactor with phosphorus removal.

Two Gram-stain-positive, non-endospore-forming actinobacteria (Ca8(T)and Ca14) were isolated from a bioreactor with extensive phosphorus removal. Based on 16S rRNA gene sequence similarity comparisons, strains Ca8(T) and Ca14 were shown to belong to the genus Gordonia and were most closely related to Gordonia hirsuta DSM 44140(T) (98.0 % sequence similarity) and Gordonia hydrophobica DSM 44015(T) (97.2 %). In comparison with the sequences of the type strains of all other species of the genus Gordonia tested, similarities were below 97 %. The quinone systems of the strains were determined to consist predominantly of MK-9H(2). The polar lipid profile for both organisms consisted of diphosphatidylglycerol, phosphatidylglycerol, phospatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannoside. Whole-organism hydrolysates contained meso-diaminopimelic acid as the diamino acid of the peptidoglycan; mycolic acids were detected as well. These chemotaxonomic traits and the major fatty acids, which were C(16 : 1)cis-9, C(16 : 0) and C(18 : 1) and tuberculostearic acid strongly supported the grouping of strains Ca8(T) and Ca14 into the genus Gordonia. The two strains showed a DNA-DNA similarity of 96 %. DNA-DNA hybridizations of strain Ca8(T) with G. hirsuta DSM 44140(T) and G. hydrophobica DSM 44015(T) resulted in values of 26.3 and 25.0 %, respectively. These results and those of the physiological and biochemical tests allowed a clear phenotypic differentiation of strains Ca8(T) and Ca14 from the most closely related species of the genus Gordonia. It is concluded that strains Ca8(T) and Ca14 represent a novel species, for which the name Gordonia phosphorivorans sp. nov. is proposed, with the type strain Ca8(T) (= DSM 45630(T) = CCUG 61533(T) = CCM 7957(T) = LMG 26648(T)).

Removal of endocrine disruptors and cytostatics from effluent by nanofiltration in combination with adsorption on powdered activated carbon.

Direct capillary nanofiltration also in combination with an upstream powdered activated carbon treatment was tested for high quality water reuse of tertiary effluent from a municipal wastewater treatment plant. Two endocrine disruptors (BPA and EE2) and two cytostatics (CytR and 5-FU) were spiked in concentrations of 1 to 2 microg/L to evaluate the process performance. In direct NF the real total removal of the micropollutants was between 5 and 40%. Adsorption to the membrane played a major role leading to a seemingly total removal between 35 and 70%. Addition of powdered activated carbon and lignite coke dust largely reduced the influence from adsorption to the membrane and increased the total removal to >95 to 99.9% depending on the PAC type and dose. The cytostatics showed already in direct NF a very high removal due to unspecified losses. Further investigations are ongoing to understand the underlying mechanism. The PAC/NF process provided a consistently high permeate quality with respect to bulk and trace organics.

Indoor environment and respiratory symptoms in children living in the Dutch-German borderland.

OBJECTIVE: To investigate the relation between indoor environmental risk factors and respiratory symptoms in 7-8-year-old children living in the Dutch-German borderland. METHODS: A nested case-control study was conducted among children participating in a large longitudinal study on respiratory health. Parents of all 781 children with respiratory complaints and an equal number of randomly selected controls were asked to complete a questionnaire, including questions on indoor environment. RESULTS: The parents of 1191 children (76.2%) participated. Past exposure to environmental (OR = 2.73, 95% CI 1.14-6.67) as well as in utero exposure (OR = 2.28, 95% CI 1.15-4.53) to tobacco smoke, use of an unvented geyser for water heating (OR = 3.01, 95% CI 1.21-7.56), long-term exposure to dampness (OR = 2.98, 95% CI 1.10-8.28) or pets (OR = 2.18, 95% CI 1.39-3.42) increased the risk of asthmatic symptoms in 7-8-year-old children. A middle or low socio-economic status also increased the risk of asthmatic symptoms. An inverse association with asthmatic symptoms was seen for wall-to-wall carpeting (OR = 0.57, 95% CI 0.33-0.95) and insulation measures (OR = 0.46, 95% CI 0.25-0.83). Except for the presence of an unvented geyser, these environmental risk factors also presented a risk for coughing symptoms in children. CONCLUSION: This study showed an increased risk of respiratory symptoms in children exposed to several indoor environmental risk factors.

[Comparison of methods for quantification of MVOC in indoor environments]. / Vergleich der Messverfahren zur Bestimmung von MVOC in Innenräumen.

For several years now, MVOC have been regarded as indicators for microbial growth in indoor environments. Until now, a direct correlation between the occurrence of microfungi and MVOC could not be shown in scientific investigations. One reason may be that different analytical methods were applied, and moreover they were not validated sufficiently. The present investigation aimed to test the comparability of both methods (Tenax adsorption/thermal desorption; charcoal adsorption/elution). It turned out that with both methods comparable results can be achieved if the technical handling of the calibration is standardized to a wide extent. Thus, highest demands have to be made on quality assurance. Therefore, it is necessary to formulate technical regulations for the quantification of MVOC.

Differences in asthma diagnosis and medication use in children living in Germany and the Netherlands.

AIM: To compare diagnosis and treatment between German and Dutch children with asthmatic symptoms at the age of 5-6 and 7-8 years, and the use of anti-asthma medication at 7-8 years of age. METHODS: Parents of 4462 children participated in two surveys, in 1995 and 1997. All 465 children identified with current asthmatic symptoms at the age of 5-6 (May 1995) or at 7-8 years of age (May 1997) were sent a third more detailed questionnaire (October 1997). RESULTS: Asthma diagnosis was more prevalent in Dutch children with current asthmatic complaints (50-60%), whereas over 90% of the German children with current asthmatic complaints had been diagnosed with bronchitis. Inhaled beta(2)-agonists were more frequently used by Dutch children compared to German children (67.3% vs. 45.6% p < 0.01) as were inhaled steroids (38.9% vs. 7.0% p < 0.01). Instead, German children more often used sodium cromoglycate or nedocromil as anti-inflammatory medication as compared with Dutch children (42.1% vs. 11.5% p < 0.01). CONCLUSIONS: Differences in diagnosis rates for asthma and bronchitis between German and Dutch children most likely result from differential labelling of complaints, and probably lead to differences in treatment practice, indicating possible undertreatment of German children with inhaled steroids.

Efficiency of nanofiltration for the elimination of steroids from water.

Steroid hormones such as the synthetic 17-alpha-ethinylestradiol often persist in the sewage treatment process and metabolites of the natural steroids like estrone from estradiol accumulate resulting in relevant amounts in the aquatic environment. Since biological removal of steroids in the sewage treatment plant is not successfully complete, physical processes such as membrane filtration may become more important. We investigated the elimination of natural and synthetic steroids by nanofiltration using a laboratory membrane reactor. Chemical analysis of 17-beta-estradiol, estrone, estriol, 17-alpha-ethinylestradiol, mestranol, diethylstilbestrol, progesterone and beta-sitosterine was performed after solid phase extraction by GC-MS with standard addition. The elimination rate depended on the nanofiltration membrane material. LFC1 membrane consisting of polyamide removed the steroids over 99% whereas PES10 membrane consisting of hydrolysed polyethersulfone was less efficient, obviously caused by different pore sizes and permeability of the membrane structure.

Infant immunization and the occurrence of atopic disease in Dutch and German children: a nested case-control study.

Our goal was to assess the role of early childhood vaccination in the occurrence of respiratory symptoms and allergic sensitization in 7-8-year-old Dutch and German children. A nested case-control study was conducted among children participating in a large longitudinal study on respiratory health, to study the relationship between vaccination (bacille Calmette-Guérin (BCG), pertussis, measles/mumps, rubella, and Haemophilus influenza type b (Hib)) and respiratory symptoms and allergic sensitization. Parents of 510 7-8-year-old children with respiratory complaints and an equal number of randomly selected children without respiratory complaints were asked to complete a questionnaire. Blood samples were collected for specific serum IgE analysis. Vaccination status was assessed through the records of the participating Municipal Health Services. No association between vaccination against pertussis, measles, rubella, or Hib and respiratory symptoms or allergic sensitization was found. For sensitization against house dust mite, BCG vaccination resulted in an increased risk (OR, 2.28; 95% CI, 1.05-4.96). Birth order was inversely associated with allergic sensitization, but was not related to respiratory symptoms. We found an association between BCG vaccination and the subsequent risk for sensitization against house dust mite. No evidence was found for an association between vaccination and respiratory symptoms. Earlier reports of an association of birth order with atopic disease were supported by the results of the present study.

Acute toxicity investigations of ester-based lubricants by using biotests with algae and bacteria.

Although ester-based lubricants are ecologically acceptable due to their good biodegradability, there are still some environmental ecotoxicological impacts that have to be considered. Information on the acute ecotoxicological behavior of lubricants is obtained in this work using several single species bioassays. In previous studies it was observed that lubricating fluids containing additives for the enhancement of their technical performance were more problematic than base fluids especially with respect to algae growth inhibition. In order to clarify the influence of additives, the anti-wear additive tri-n-butyl phosphate was tested. It was very toxic to algae though not to bacteria. Additionally, a mixture of this additive with a base fluid is characterized. Despite the high toxicity of the single additive, the water extract of the mixture of tri-n-butyl phosphate with hydraulic base fluid caused almost no toxicity. Therefore, tri-n-butyl phosphate cannot explain the effect observed for the toxicity of water extracts of the commercially available lubricants.

Effect of sample preparation on the in vitro genotoxicity of a light curable glass ionomer cement.

The glass ionomer cement Vitrebond showed a clear genotoxic effect in the in vitro Mammalian Cell Gene Mutation Test (HPRT Test) with CHO cells as well as in the bacterial umu-test with Salmonella typhimurium TA1535/pSK1002. Both DMSO and Ham's F12 cell culture medium extracts according to ISO 10993-12 (Biological evaluation of medical devices-Part 12: sample preparation and reference materials, Geneva, Switzerland) exhibit a clear genotoxic effect in the umu-test. The effect is independent of the extraction volume in a range from 0.5 to 4 ml Ham's F12 cell culture medium. Subsequent extractions of Vitrebond showed no significant difference in the genotoxic response although weight loss and content of 2-hydroxyethyl-methacrylate dropped significantly. In vivo conditions of Vitrebond were simulated by extractions with artificial and collected human saliva. These extracts showed a clear genotoxic effect in the umu-test, even if only a few seconds of extraction time were applied. In conclusion, sample preparations for genotoxicity testing according to ISO 10993-12 reflect the in vivo conditions of Vitrebond applications. This seems to be mostly due to the hydrophilic nature of the genotoxic ingredients.

Quality assurance and good laboratory practice in the mycological laboratory--compilation of basic techniques for the identification of fungi.

The identification of microfungi by morphology is generally regarded to be very difficult, as traits used for delimitation of species within a genus often show minute differences that can only be reliably evaluated by experienced mycologists. Different strains of one species can slightly vary in their morphology. This is often used as an argument by investigators with rather limited experience that identification on a morphological basis is not reliable and leads to inconsistent results. This paper compiles essential methods and techniques contributing to improved identification concerning (i) isolation from mixed cultures, (ii) inoculation and incubation, (iii) usage of media, (iv) identification and usage of determination keys, (v) preparation of microscopic slides, (vi) microscopy, and (vii) storage of isolates. The application of chemotaxonomy and sequencing for routine identification of airborne microfungi is shortly evaluated. The techniques compiled here can improve routine determination work considerably and will lead to much more accuracy and consistency of results. An improved quality assurance of applied fungal identification in the field of indoor and occupational hygiene is a prerequisite to find possible correlations between exposure on the one hand and health effects on the other.

Selective enrichment and characterization of a phosphorus-removing bacterial consortium from activated sludge.

Under alternating aerobic/anaerobic conditions and without additional carbon sources, a bacterial consortium consisting initially of 18 bacterial strains was obtained in a sequence batch reactor. The phosphorus removal capability could only be maintained using sterile filtrate of activated sludge as medium. The addition of calcium and magnesium salts, as well as vitamins and trace elements, to autoclaved sterile filtrate of activated sludge was not sufficient to achieve stable phosphorus removal. A further enrichment by subcultivation on solid, agar, freezing, and shortening of the aerobic and anaerobic phases led to a defined bacterial consortium consisting of four strains. On the basis of physiological and chemotaxonomic characterization, and partial 16S rRNA sequencing, one of the organisms was identified as Delftia acidovorans. A further isolate belonged to the Bacillus cereus group, and the third isolate was identified as Microbacterium sp.. The remaining strain seems to represent a new genus within the Flavobacteriaceae. Under continuous chemostat conditions, this consortium was able to remove up to 9.6 mg P/l phosphate in the aerobic phase and released up to 8.5 mg/l in the anaerobic phase. Up to 25 mg P-polyphosphate/g dry mass was stored under aerobic conditions.

Denitrification of groundwater with methane as sole hydrogen donor.

It was examined, whether methane can be used as hydrogen donor for an in situ denitrification of groundwater. It is demonstrated, that groundwater can serve as liquid medium and that the denitrification can occur at 10 degrees C. Efforts to enrich methanotrophic bacteria under anoxic conditions have not been successful. No methane oxidation occurred in the absence of oxygen. For this reason, the denitrification with methane must be performed in a two-stage process with aerobic methanotrophic bacteria producing metabolites, that are used as hydrogen donor by non-methanotrophic bacteria in anoxic areas. This kind of indirect denitrification was proved by quantifying nitrogen and nitrous oxide in enrichment cultures that were not stirred or shaken. Large numbers of non-methanotrophic bacteria being able to denitrify with methanol, acetate or proteins as sole hydrogen donor were enriched besides the methanotrophic bacteria under these conditions.

[Interreg II--European regional study of chronic respiratory tract diseases in children in the Germany-Netherlands border zone]. / Interreg II--Euregionale Untersuchungen zu chronischen Atemwegserkrankungen bei Kindern im deutsch-niederländischen Grenzraum.

AIM OF THE STUDY: In Westelijke Mijnstreek (WM), Midden-Limburg (ML), both The Netherlands, and Kreis Heinsberg (HS), Germany, the relationship between quality of the indoor environment and respiratory symptoms in children was studied, finally resulting in proposals for prevention of respiratory symptoms in the three regions. This paper presents the results of the first part of the study, investigating prevalences of respiratory complaints. METHODS AND RESULTS: A brief questionnaire was used to document prevalences of respiratory complaints, physicians diagnoses and treatment in about 7,000 children 5-6 resp. 7-8 years of age from HS, ML and WM, in 1995 and 1997. Prevalences of respiratory symptoms differed slightly between the regions, but there were differences in physician-diagnosed asthma and bronchitis and treatment of the children. PERSPECTIVES: The brief questionnaire is a useful instrument to get an overview on respiratory symptoms. Since 1995, it is used by several German Municipal Health Services for Public Health Reports.

Microcosms-experiments to assess the potential for natural attenuation of contaminated groundwater.

Groundwater samples from six wells of a former gas plant site were characterised using chemical, microbial and ecotoxicological methods. Degradation studies were performed in batch-culture under aerobic conditions with the groundwater samples containing their autochthonous microflora and original contaminant mixture. The highest O2-consumption (3 mmol 100 ml-1), combined with BTEX (8.3 mg l-1) and naphthalene (171.3 mg l-1) degradation, as well as formation of organic acids was found after N- and P-supplementation with the highest contaminated groundwater sample. The other highly polluted groundwater sample showed no activity obviously because of the toxicity of some compounds. The major part of the PAHs and BTEX was eliminated in the assays with the low contaminated groundwater samples. The results indicate that the microbial degradation capacity and thereby the natural attenuation capacity in each groundwater differ and cannot be assessed simply by chemical, microbial and toxicological data. Additionally activity tests with authentic groundwater samples with and without nutrient supplementation are recommended.