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Biomedical subjects

Per H Nielsen

Publications and source records attributed to Per H Nielsen.

16 recordsLinked to original sources

Abundance and ecophysiology of Defluviicoccus spp., glycogen-accumulating organisms in full-scale wastewater treatment processes.

The activity of glycogen-accumulating organisms (GAOs) in enhanced biological phosphorus removal (EBPR) wastewater treatment plants has been proposed as one cause of deterioration of EBPR. Putative GAOs from the Alphaproteobacteria, Defluviicoccus spp. (including D. vanus), were studied in full-scale EBPR plants to determine their distribution, abundance and ecophysiology. Fluorescence in situ hybridization (FISH) demonstrated that Defluviicoccus spp. were generally low in abundance; however, in one plant surveyed, Cluster 2 Defluviicoccus constituted 9 % of all Bacteria. FISH combined with microautoradiography revealed that both Cluster 1 and Cluster 2 Defluviicoccus were capable of taking up a narrow range of substrates including acetate, propionate, pyruvate and glucose under anaerobic and aerobic conditions. Formate, butyrate, ethanol and several other substrates were not taken up. Cluster 2 Defluviicoccus demonstrated a phenotype consistent with the current metabolic model for GAOs--anaerobic assimilation of acetate and reduction to polyhydroxyalkanoates (PHA) using the glycolytic pathway, and aerobic consumption of PHA. Polyphosphate-accumulating organisms (PAOs, 'Candidatus Accumulibacter phosphatis') and other putative GAOs ('Candidatus Competibacter phosphatis') co-existed in two plants with Cluster 2 Defluviicoccus, but in both plants, the latter organisms were more abundant. Thus Cluster 2 Defluviicoccus can be relatively abundant and could be carbon competitors of PAOs and other GAOs in EBPR plants.

Acetates↗

Unexplained week-to-week variation in BNP and NT-proBNP is low in chronic heart failure patients during steady state.

BACKGROUND: The usefulness of brain-natriuretic-peptide (BNP) and N-terminal-pro-brain-natriuretic-peptide (NT-proBNP) for monitoring of chronic heart failure (CHF) patients has been questioned because of high levels of unexplained variation. AIMS: Week-to-week total variance (CV(T)), unexplained variation (CV(I)), reference change values (RCV), index of individualities (IOI) and number of samples (N) with week-to-week intervals needed to estimate the underlying homeostatic set point (+/-15%) for BNP and NT-proBNP were calculated in pre-specified stable CHF patients. METHODS AND RESULTS: We measured plasma concentrations of BNP and NT-proBNP, clinical and laboratory variables in 20 CHF patients with a 7-days interval. Only patients considered to be in steady state were included. The CV(I) was 15% (BNP) and 8% (NT-proBNP). CV(T) was 16% (BNP) and 8% (NT-proBNP) and RCV was 43% (BNP) and 23% (NT-proBNP). IOI was 0.14 for BNP and 0.03 for NT-proBNP and N was 1 for BNP and 1 for NT-proBNP. CONCLUSIONS: Our data demonstrate that unexplained variation of BNP and NT-proBNP is low in CHF patients during steady state, which is a prerequisite for the use of these peptides for monitoring of the disease.

Aged↗

Cohn's Crenothrix is a filamentous methane oxidizer with an unusual methane monooxygenase.

135 years ago Ferdinand Cohn, the founder of bacteriology, microscopically observed a conspicuous filamentous bacterium with a complex life cycle and described it as Crenothrix polyspora. This uncultured bacterium is infamous for mass developments in drinking water systems, but its phylogeny and physiology remained unknown. We show that C. polyspora is a gammaproteobacterium closely related to methanotrophs and capable of oxidizing methane. We discovered that C. polyspora encodes a phylogenetically very unusual particulate methane monooxygenase whose expression is strongly increased in the presence of methane. Our findings demonstrate a previously unrecognized complexity of the evolutionary history and cell biology of methane-oxidizing bacteria.

Bacterial Proteins↗

Meganema perideroedes gen. nov., sp. nov., a filamentous alphaproteobacterium from activated sludge.

An industrial wastewater treatment plant at Grindsted, Denmark, has suffered from bulking problems for several years caused by filamentous bacteria. Five strains were isolated from the sludge by micromanipulation. Phylogenetic analysis of the 16S rRNA gene sequences showed that the strains formed a monophyletic cluster in the Alphaproteobacteria, and they were phenotypically different from their closest relatives and from all hitherto known filamentous bacteria described (closest relative Brevundimonas vesicularis ATCC 11426(T), 89.8 % sequence similarity). In pure culture, the cells (1.5-2.0 microm) in filaments are Gram-negative and contain polyphosphate and polyhydroxyalkanoates. The optimum temperature for growth is 30 degrees C and the strains grow in 2 % NaCl and are oxidase- and catalase-positive. Ubiquinone 10 is the major quinone. The major fatty acid (C(18 : 1)omega7c) and smaller amounts of unsaturated fatty acids, 3-hydroxy fatty acids with a chain length of 16 and 18 carbon atoms and small amounts of 10-methyl-branched fatty acids with 18 carbon atoms (C(19 : 0) 10-methyl) affiliated the strains with the Methylobacterium/Xanthobacter group in the Alphaproteobacteria. The G+C content of the DNA is 42.9 mol% (for strain Gr1(T)). The two most dissimilar isolates by 16S rRNA gene comparison (Gr1(T) and Gr10; 97.7 % identical) showed 71.5 % DNA-DNA relatedness. Oligonucleotide probes specific for the pure cultures were designed for fluorescence in situ hybridization and demonstrated that two filamentous morphotypes were present in the Grindsted wastewater treatment plant. It is proposed that the isolates represent a new genus and species, Meganema perideroedes gen. nov., sp. nov. The type strain of Meganema perideroedes is strain Gr1(T) (=DSM 15528(T)=ATCC BAA-740(T)).

Alphaproteobacteria↗

Ecophysiology of a group of uncultured Gammaproteobacterial glycogen-accumulating organisms in full-scale enhanced biological phosphorus removal wastewater treatment plants.

The presence of glycogen-accumulating organisms (GAOs) in enhanced biological phosphorus removal (EBPR) plants can seriously deteriorate the biological P-removal by out-competing the polyphosphate-accumulating organisms (PAOs). In this study, uncultured putative GAOs (the GB group, belonging to the Gammaproteobacteria) were investigated in detail in 12 full-scale EBPR plants. Fluorescence in situ hybridization (FISH) revealed that the biovolume of the GB bacteria constituted 2-6% of total bacterial biovolume. At least six different subgroups of the GB bacteria were found, and the number of dominant subgroups present in each plant varied between one and five. Ecophysiological investigations using microautoradiography in combination with FISH showed that, under aerobic or anaerobic conditions, all subgroups of the GB bacteria could take up acetate, pyruvate, propionate and some amino acids, while some subgroups in addition could take up formate and thymidine. Glucose, ethanol, butyrate and several other organic substrates were not taken up. Glycolysis was essential for the anaerobic uptake of organic substrates. Polyhydroxyalkanoates (PHA) but not polyphosphate (polyP) granules were detected in all GB bacterial cells. Polyhydroxyalkanoate formation after anaerobic uptake of acetate was confirmed by measuring the increase in fluorescence intensity of PHA granules inside GB bacterial cells after Nile blue staining. One GB subgroup was possibly able to denitrify, and several others were able to reduce nitrate to nitrite. PAOs were also enumerated by FISH in the same treatment plants. Rhodocyclus-related PAOs and Actinobacteria-related PAOs constituted up to 7% and 29% of total bacterial biovolume respectively. Rhodocyclus-related PAOs always coexisted with the GB bacteria and showed many physiological similarities. Factors of importance for the competition between the three groups of important bacteria in EBPR plants are discussed.

Acetates↗

The in situ physiology of Skermania piniformis in foams in Australian activated sludge plants.

The in situ physiology of the filamentous bacterium Skermania piniformis frequently seen in activated sludge foams in Australia was investigated. An oligonucleotide probe, Spin1449, targeting the 16S rRNA of S. piniformis was designed for its identification by fluorescence in situ hybridization (FISH), validated with pure cultures and applied successfully to foam samples from two geographically distant Australian plants. While filaments of this bacterium appeared to be comparatively hydrophobic, the organism had no clear preference for hydrophobic or hydrophilic substrates. In both foams examined using microautoradiography (MAR), filaments selectively took up substrates under aerobic and anoxic (NO(3) (-)) but not anaerobic or anoxic (NO(2) (-)) conditions. Skermania piniformis assimilated oleic acid, palmitic acid, glycerol and glycine. Ectoenzyme activities detected suggest that S. piniformis has an ability to assimilate a greater range of substrates than might be concluded from the MAR data obtained here. Based on the substrate uptake data presented here, an anaerobic selector may work for controlling S. piniformis in activated sludge systems.

Actinobacteria↗

Detection of activity among uncultured Actinobacteria in a drinking water reservoir.

The abundance, identity and activity of uncultured Bacteria and Actinobacteria present in a drinking water reservoir (North Pine Dam, Brisbane, Australia) were determined using a combination of fluorescence in situ hybridization (FISH) alone or with catalysed reporter deposition (CARD-FISH) with microautoradiography. The CARD-FISH technique was modified relative to previous described procedures and performed directly on gelatine cover slips in order to allow simultaneous combination with microautoradiography. Almost twofold higher numbers of microorganisms could be identified as either Bacteria or Actinobacteria using the CARD-FISH technique as compared with the traditional FISH technique. A combination of FISH or CARD-FISH with microautoradiography showed generally higher activity among the Actinobacteria than among all Bacteria. Another important observation was that many cells within the FISH-negative populations of both Actinobacteria and Bacteria were actively assimilating thymidine. Thus, great care should be taken when extrapolating the active fraction of a prokaryotic community to be equivalent to the FISH-detectable population in such environments. Bacterial groups within Actinobacteria produce the odours geosmin and 2-methylisoborneol, which lower the quality of surface water when used for drinking. The results indicate that combined microautoradiography and CARD-FISH may serve as an effective tool when studying identity and activity of microorganisms within freshwater environments.

Actinobacteria↗

Development of methods for usability evaluations of EHR systems.

Developing electronic health record (EHR) systems in Denmark is an on going, iterative process, where also a maturation process for clinical use should be considered. Convincing methodology for collecting and incorporating in the soft- and hardware knowledge and robustness for the clinical environments is not on hand. A way to involve the clinicians in the development process is conducting usability evaluations. The complexity of the clinical use of the systems is difficult to transmit to a usability laboratory, and due to ethical issues a traditional field study can be impossible to carry out. The aim of this study has been to investigate how it is possible to identify usability problems in an EHR system by combining methods from laboratory tests and field studies. The methods selected for the test design are: the think aloud method, video and screen recording, debriefing, a scenario based on an authentic patient record, and testing on the normal production system. The reliability and validity of the results is increased due to the application of method- and data-triangulation. The results of the usability evaluation include problems in the categories: system response time, GUI-design, functionality, procedures, and error messages. The problems were classified as cosmetic, severe, or critical according to a rating scale. The experience with each method is discussed. It is concluded that combining methods from laboratory test and field study makes it possible to identify usability problems. There are indications that some of the usability problems only occurred due to the establishment of an authentic scenario.

Denmark↗

Linking microbial community structure with function: fluorescence in situ hybridization-microautoradiography and isotope arrays.

The ecophysiology of microorganisms has been at the heart of microbial ecology since its early days, but only during the past decade have methods become available for cultivation-independent, direct identification of microorganisms in complex communities and for the simultaneous investigation of their activity and substrate uptake patterns. The combination of fluorescence in situ hybridization (FISH) and microautoradiography (MAR) is currently the most widely applied tool for revealing physiological properties of microorganisms in their natural environment with single-cell resolution. For example, this technique has been used in wastewater treatment and marine systems to describe the functional properties of newly discovered species, and to identify microorganisms responsible for key physiological processes. Recently, the scope of FISH-MAR was extended by rendering it quantitative and by combining it with microelectrode measurements or stable isotope probing. Isotope arrays have also been developed that exploit the parallel detection offered by DNA microarrays to measure incorporation of labelled substrate into the rRNA of many community members in a single experiment.

Archaea↗

Substrate specificity of the bovine serum amine oxidase and in situ characterisation of aminoaldehydes by NMR spectroscopy.

The oxidation of spermidine or homospermidine with bovine serum amine oxidase (BSAO) was monitored in situ, using proton nuclear magnetic resonance spectroscopy in water with 10% D(2)O. NMR assignments were performed by spin decoupling and COSY spectra or by comparison with data from synthetic aminoaldehydes. The results represent the first in situ characterisation of the highly reactive aminoaldehydes and showed oxidation at the N(1) amino group of spermidine and homospermidine. Comparison of homospermidine with a variety of substrates revealed that among straight chain di- and polyamines both an aminopropyl group and two primary amino groups separated by seven (norspermidine) or eight (spermidine) carbon atoms were required for optimal substrate ability. However, highest activity was seen with the substrate N-(4-aminobutyl)hexahydropyrimidine, showing that the substrate channel of BSAO has a dual substrate preference, with moderately bulky substituents at the distal end of a diamine contributing equally well as an alkyl amino group. Cytotoxic investigations of a variety of substrates for BSAO, confirmed previous results, that cytotoxicity is primarily linked to polyamines encompassing the aminopropyl moiety. No acrolein was observed at any time during the oxidation showing that it reacts very fast with available amino groups forming a variety of derivatives.

Aldehydes↗

"Microthrix parvicella", a filamentous bacterium causing bulking and foaming in activated sludge systems: a review of current knowledge.

This review summarizes the microbiology and physiology of "Microthrix parvicella" and the methods of its growth control in activated sludge wastewater treatment plants. This filamentous bacterium is of high interest because of its worldwide involvement in severe bulking and foaming at wastewater treatment plants. We present a critical analysis of physiological and kinetic data on "M. parvicella" and discuss its growth and storage abilities in various environments with the aim of understanding the strategies of this organism to successfully compete with other bacteria in activated sludge. Additionally, this review elaborates on research needs for defining reliable control strategies of bulking and foaming based on key features of "M. parvicella".

Actinobacteria↗

[Establishment of a nurse-led heart failure clinic. Design and baseline data from the first two years].

INTRODUCTION: Previous studies on the effect of heart failure clinics have shown a reduction in hospitalizations, reduced cost and improved quality of life. MATERIAL AND METHODS: We report on the establishment of a heart failure clinic and its impact on heart failure hospitalizations. Since September 1, 1999, a heart failure clinic has been operated at the Frederiksberg University Hospital. The clinic was designed with both a diagnostic and a therapeutic unit. The diagnostic unit offers open access to all patients with suspected heart failure, either through referral from general practitioners or from the medical departments of the hospital. In case of confirmed systolic heart failure, the patient is referred to the therapeutic unit. RESULTS: During the two years of operation, a total of 510 patients were registered in a newly established database (HJERTER+). Of these, 352 (69%) were found to have clinical evidence of heart failure, and 304 (86% of the heart failure patients) had left ventricular (LV) systolic dysfunction (ejection fraction < or = 0.45). Before referral, most patients with LV dysfunction had been treated with no or insufficient dose of ACE inhibitor (55%), and one third received beta-blockers (32%). During the two years of operation of the clinic, there was a 23% decline in heart failure related hospital admissions to the department of cardiology. DISCUSSION: Patients with systolic heart failure are not always optimally treated. The establishment of a heart failure clinic offering the combination of diagnosing and managing congestive heart failure appears to be effective both in terms of therapy optimization and with respect to a concomitant decline in hospitalization for heart failure.

Aged↗

Lipase and protease extraction from activated sludge.

In the process of wastewater treatment hydrolysis of polymeric substances is the first and rate-limiting step. A closer study of the enzymes catalysing these reactions is essential for a better understanding of the microbial activity in the wastewater treatment process. Therefore, development of gentle and efficient enzyme extraction methods from environmental samples is very important. In this study we present a method for the extraction of lipases and proteases from activated sludge using the non-ionic detergent Triton X-100, EDTA, and cation exchange resin (CER), alone or in combination for the extraction of lipases and proteases from activated sludge. The sludge was continuously stirred in the presence of either buffer alone or in the presence of detergent and/or chelating agents. In all cases, a marked reduction in floc size was observed upon continuous stirring. However, no lipase activity and negligible protease activity was extracted in the presence of buffer alone, indicating that enzyme extraction was not due to shear force alone. The highest lipase activity was extracted using 0.1% Triton X-100 above which the activity was gradually decreasing. For proteases, the highest activity was obtained in the presence of 0.5% Triton X-100 and no decrease in activity was observed. Differences observed in the extraction efficiency of the two enzymes indicate the need for optimisation of the extraction process for the different enzymes or the extracellular polymeric substances from activated sludge.

Cation Exchange Resins↗

Long-term population dynamics and in situ physiology in activated sludge systems with enhanced biological phosphorus removal operated with and without nitrogen removal.

Quantitative fluorescence in situ hybridization (FISH) and the combination of FISH with microautoradiography (MAR) were used in order to study the long-term population dynamics (2.5 years) and the in situ physiology in two parallel activated sludge pilot systems with enhanced biological phosphorus removal (EBPR). The two systems received the same influent wastewater, but were differently operated (with and without nitrogen removal, respectively). Both systems showed a significant P removal that increased when different substrates (phosphorus (P), acetate and glucose, respectively) were added to the influent wastewater. Rhodocyclus-related bacteria were present in both systems in significant numbers (ranging from 4 to 28%) throughout the whole period. This supports the hypothesis that these bacteria occur in significant numbers in different types of well-operating EBPR activated sludge processes. However, we observed a lower correlation (< 0.5) for the amount of Rhodocyclus-related bacteria to the P content in activated sludge than previous studies (> 0.9). The Actinobacteria were the only additional group of bacteria which showed a similar degree of correlation to the P content in activated sludge as the Rhodocyclus-related bacteria--but only for the system without nitrogen removal. Significant amounts (< or = 12%) of glycogen-accumulating bacteria (GAOs) were detected in the system with nitrogen removal (but not in the other system), but had no, in contrast to previous observations, apparent negative effect on the overall EBPR performance. FISH-MAR indicated that a significant part of the Betaproteobacteria (part of them identified as Rhodocyclus-related bacteria) as well as the Actinobacteria were able to take up 33Pi, [3H]-acetate and [3H]-glucose under anaerobic-aerobic conditions. The contribution of anoxic 33Pi uptake under alternating anaerobic-anoxic conditions was significantly lower. Interestingly, not all Rhodocyclus-related bacteria showed uptake of these three radioactive substrates. This may be due to differences in metabolic state, physiological potential or genotype, not detectable by the present probe set for Rhodocyclus-related bacteria. Comparison of the 33Pi, [3H]-acetate and [3H]-glucose uptake by activated sludge after different fixation and incubation procedures showed that a part of the observed 33Pi uptake may have been caused by a combination of a biological and chemical or biologically induced chemical P adsorption.

Archaea↗

An integrated approach to diagnosis and therapeutic management of patients with systolic heart failure in the Copenhagen metropolitan area.

BACKGROUND: Studies on the effect of heart failure clinics have been encouraging and convincing with respect to reduced hospitalizations, improved quality of life, and reduced cost. However, previous results have been derived from patient populations either recently discharged and with specific exclusion criteria or from heart transplant lists. We here report the establishment of a heart failure clinic that offers an integrated approach to both diagnosing and managing heart failure patients, the baseline characteristics of the first 283 patients treated, and the impact on heart failure hospitalizations during establishment. METHODS: Since September l, 1999, a heart failure clinic has been in operation at Frederiksberg University Hospital. The clinic was designed with both a diagnostic and a therapeutic unit. The diagnostic unit offers open access to all patients with suspected heart failure, either through referral from general practitioners or from the medical departments of the hospital. In the case of confirmed systolic heart failure, the patient is referred to the therapeutic unit for (1) uptitration and optimization of medical therapy and thorough information and education. In addition, the unit offers a rapid access track and the possibility of intravenous diuretic treatment. RESULTS: During the 21 months of operation, a total of 460 patients were referred to the clinic. Of these, 320 (70%) were found to have clinical evidence of heart failure, and 283 (88% of the patients with heart failure) had left ventricular systolic dysfunction (defined by an ejection fraction of < or =45%). These patients were seen in the clinic for education, control of fluid retention, and uptitration of medications. These patients with left ventricular dysfunction were predominantly elderly (aged 72.7 +/- 10.7 y) and most often male (73%). Ischemic heart disease was the most common etiology (55%). Before referral, most patients with left ventricular dysfunction had been treated with none or only low-dose angiotensin-converting enzyme inhibitor (55%), and fewer than one third received beta-blockers (29%). During the first year of operation of the clinic, there was a 23% decline of heart failure-related hospital admissions to the department of cardiology (from 396 to 306). CONCLUSIONS: Patients with systolic heart failure are not always optimally treated. The establishment of a heart failure clinic, which offers the combination of diagnosing and managing congestive heart failure, appears to be efficient both in terms of therapy optimization and with respect to a concomitant decline in hospitalization for heart failure.

Adolescent↗

Phylogenetic identification and substrate uptake patterns of sulfate-reducing bacteria inhabiting an oxic-anoxic sewer biofilm determined by combining microautoradiography and fluorescent in situ hybridization.

We simultaneously determined the phylogenetic identification and substrate uptake patterns of sulfate-reducing bacteria (SRB) inhabiting a sewer biofilm with oxygen, nitrate, or sulfate as an electron acceptor by combining microautoradiography and fluorescent in situ hybridization (MAR-FISH) with family- and genus-specific 16S rRNA probes. The MAR-FISH analysis revealed that Desulfobulbus hybridized with probe 660 was a dominant SRB subgroup in this sewer biofilm, accounting for 23% of the total SRB. Approximately 9 and 27% of Desulfobulbus cells detected with probe 660 could take up [(14)C]propionate with oxygen and nitrate, respectively, as an electron acceptor, which might explain the high abundance of this species in various oxic environments. Furthermore, more than 40% of Desulfobulbus cells incorporated acetate under anoxic conditions. SRB were also numerically important members of H(2)-utilizing and (14)CO(2)-fixing microbial populations in this sewer biofilm, accounting for roughly 42% of total H(2)-utilizing bacteria hybridized with probe EUB338. A comparative 16S ribosomal DNA analysis revealed that two SRB populations, related to the Desulfomicrobium hypogeium and the Desulfovibrio desulfuricans MB lineages, were found to be important H(2) utilizers in this biofilm. The substrate uptake characteristics of different phylogenetic SRB subgroups were compared with the characteristics described to date. These results provide further insight into the correlation between the 16S rRNA phylogenetic diversity and the physiological diversity of SRB populations inhabiting sewer biofilms.

Aerobiosis↗