Lifetime and Performance Assessment of Commercial Electric Double-Layer Capacitors Based on Cover Layer Formation.

The lifetime and performance of energy storage systems are essential characteristics for a major success of clean energy innovations, especially regarding the automotive sector. In this area, electric double-layer capacitors represent the cutting edge of nonfaradaic, high power, and long lifetime energy storages. Usually, degradation is neglected while operating or it is assessed by referring to simple rules of thumb. To better address the aging effects, commercial specimens have been investigated in-depth. The works can be split into two parts: first, extensive accelerated aging for more than 3 years to statistically analyze the degradation trend and significantly improve the current rules. Second, cell opening and surface characterization of the electrodes to gain a profound understanding of the ongoing processes and to correlate aging mechanisms to the statistics of the first part. It is found that a prominent cover layer forms during degradation on the positive electrode scaling with lost capacitance. The used methods for in-depth characterization include microscopy, X-ray diffraction, and thermogravimetric analysis with subsequent mass spectrometry. The newly formed layer consists of poly(tetrafluoroethylene) (also known as Teflon) that is still passable by charge carriers, albeit with a longer time constant. Additionally, the negative electrode shows corrosion and loss of contact. The composition of the cover layer has not been known yet. Thus, materials and especially electrolyte development can benefit from the results.

Molecular characterization of endophytic fungi associated with the roots of Chenopodium quinoa inhabiting the Atacama Desert, Chile.

Plant roots can be highly colonized by fungal endophytes. This seems to be of particular importance for the survival of plants inhabiting stressful habitats. This study focused on the Identification of the fungal endophytic community associated with the roots of quinoa plants (Chenopodium quinoa) growing near the salt lakes of the Atacama Desert, Chile. One hundred endophytic fungi were isolated from healthy quinoa roots, and the internal transcribed spacer (ITS) region was sequenced for phylogenetic and taxonomic analysis. The isolates were classified into eleven genera and 21 distinct operational taxonomic units (OTUs). Despite a relatively high diversity of root endophytic fungi associated with quinoa plants, the fungal community was dominated by only the Ascomycota phyla. In addition, the most abundant genera were Penicillium, Phoma and Fusarium, which are common endophytes reported in plant roots. This study shows that roots of C. quinoa harbor a diverse group of endophytic fungi. Potential roles of these fungi in plant host tolerance to stressful conditions are discussed.

Influence of a low dosage of clopidogrel on platelet function in cats as measured by the platelet function analyser PFA-100 and the multiplate analyser.

The antiplatelet drug clopidogrel is widely used for prophylaxis of arterial thromboses in cats in a standard dosage of 18.75mg per cat once daily. The aim of the study was to verify if a reduced daily dose of 10mg clopidogrel per cat has a similar antiplatelet effect as the standard dosage. Platelet function was measured with the platelet function analyser PFA-100® and a novel impedance aggregometer. Suitability of the platelet function analyser was tested in citrated blood samples of 59 healthy cats and reference ranges were established. In addition, agonist concentrations for impedance aggregometry were optimised. In the main experiment two groups of 6 healthy cats received clopidogrel either in a dosage of 10 or 18.75mg per cat over a period of seven days. Analyses were performed on days 1, 2, 3, 5, and 7. In comparison to baseline both clopidogrel dosages showed an inhibitory effect on results of the platelet function analyser and velocity of ADP-induced platelet aggregation. Values at all times were different from baseline, with the exception of day 1 in cats receiving 10mg clopidogrel where the closure time of the platelet function analysis and part of ADP-induced aggregation did not show a significant difference. Significant differences were not found between the two doses. In conclusion, our study indicates that 10mg clopidogrel per day may be as effective as 18.75mg although the latter may be advantageous as an initial loading dosage to achieve effective levels more rapidly.

Yamamoto New Scalp Acupuncture, Applied Kinesiology, and Breathing Exercises for Facial Paralysis in a Young Boy Caused by Lyme Disease-A Case Report.

The case study reports on the effect of pharmacological, complementary, and alternative medicine including YNSA, Applied Kinesiology, and respiratory exercises in a 9-year-old boy with facial paralysis. The boy suffered from borreliosis and one-sided facial paralysis that occurred 3.5 weeks after being bitten by a tick and persisted despite 4 weeks of medication with antibiotics. In the first treatment, muscle function as assessed by the coachman׳s test was normalized, and improvement in the facial paralysis was observed. Within 8 additional treatments over a period of 2 months, the boy showed complete recovery. The case shows a multimodal approach to facial paralysis integrating pharmacological treatment and CAM including YNSA, Applied Kinesiology, and breathing exercises.

VOC emissions of Grey poplar leaves as affected by salt stress and different N sources.

Nitrogen nutrition and salt stress experiments were performed in a greenhouse with hydroponic-cultured, salt-sensitive Grey poplar (Populus x canescens) plants to study the combined influence of different N sources (either 1 mm NO(3) (-) or NH(4)(+)) and salt (up to 75 mm NaCl) on leaf gas exchange, isoprene biosynthesis and VOC emissions. Net assimilation and transpiration proved to be highly sensitive to salt stress and were reduced by approximately 90% at leaf sodium concentrations higher than 1,800 microg Na g dry weight (dw)(-1). In contrast, emissions of isoprene and oxygenated VOC (i.e. acetaldehyde, formaldehyde and acetone) were unaffected. There was no significant effect of combinations of salt stress and N source, and neither NO(3)(-) or NH(4)(+) influenced the salt stress response in the Grey poplar leaves. Also, transcript levels of 1-deoxy-d-xylulose 5-phosphate reductoisomerase (PcDXR) and isoprene synthase (PcISPS) did not respond to the different N sources and only responded slightly to salt application, although isoprene synthase (PcISPS) activity was negatively affected at least in one of two experiments, despite high isoprene emission rates. A significant salt effect was the strong reduction of leaf dimethylallyl diphosphate (DMADP) content, probably due to restricted availability of photosynthates for DMADP biosynthesis. Further consequences of reduced photosynthetic gas exchange and maintaining VOC emissions are a very high C loss, up to 50%, from VOC emissions related to net CO(2) uptake and a strong increase in leaf internal isoprene concentrations, with maximum mean values up to 6.6 microl x l(-1). Why poplar leaves maintain VOC biosynthesis and emission under salt stress conditions, despite impaired photosynthetic CO(2) fixation, is discussed.

Interaction of nitrogen nutrition and salinity in Grey poplar (Populus tremula x alba).

Salinity represents an increasing environmental problem in managed ecosystems. Populus spp. is widely used for wood production by short-rotation forestry in fertilized plantations and can be grown on saline soil. Because N fertilization plays an important role in salt tolerance, we analysed Grey poplar (Populus tremula x alba, syn. Populus canescens) grown with either 1 mM nitrate or ammonium subjected to moderate 75 mM NaCl. The impact of N nutrition on amelioration of salt tolerance was analysed on different levels of N metabolism such as N uptake, assimilation and N (total N, proteins and amino compounds) accumulation. Na concentration increased in all tissues over time of salt exposure. The N nutrition-dependent effects of salt exposure were more intensive in roots than in leaves. Application of salt reduced root increment as well as stem height increase and, at the same time, increased the concentration of total amino compounds more intensively in roots of ammonium-fed plants. In leaves, salt treatment increased concentrations of total N more intensively in nitrate-fed plants and concentrations of amino compounds independently of N nutrition. The major changes in N metabolism of Grey poplar exposed to moderate salt concentrations were detected in the significant increase of amino acid concentrations. The present results indicate that N metabolism of Grey poplar exposed to salt performed better when the plants were fed with nitrate instead of ammonium as sole N source. Therefore, nitrate fertilization of poplar plantations grown on saline soil should be preferred.

Nitrogen uptake and metabolism in Populus x canescens as affected by salinity.

External salinization can affect different steps of nitrogen (N) metabolism (ion uptake, N assimilation, and amino acid and protein synthesis) depending on the inorganic N source. Here, we assessed the net uptake of N supplied as nitrate or ammonium and N assimilation (combining metabolite analyses with molecular biological approaches) in grey poplar (Populus x canescens) plants grown under saline (75 mM NaCl) and control conditions. The specific (micromol N g(-1) dry weight fine roots h(-1)) and total plant (micromol N per plant h(-1)) N net uptake rates, total plant N content, total plant biomass and total leaf protein concentration were reduced under saline conditions when plants were supplied with ammonium. In both nutritional groups, salt treatment caused pronounced accumulation of soluble N compounds in the leaves. The mRNAs of genes coding for enzymes catalyzing rate-limiting steps of both proline synthesis and degradation (delta-1-pyrroline-5-carboxylate synthase and proline dehydrogenase) as well as for NADH-dependent glutamate synthase were accumulated under saline conditions. Whereas under control conditions the plant N status seemed to be superior when ammonium was supplied, the N balance of ammonium-fed plants was more severely affected by salt stress than that of plants supplied with nitrate. Possible metabolic implications of stress-related accumulation of particular amino acids are discussed.

Sequence of the 50-kb conjugative multiresistance plasmid pRE25 from Enterococcus faecalis RE25.

The complete 50,237-bp DNA sequence of the conjugative and mobilizing multiresistance plasmid pRE25 from Enterococcus faecalis RE25 was determined. The plasmid had 58 putative open reading frames, 5 of which encode resistance to 12 antimicrobials. Chloramphenicol acetyltransferase and the 23S RNA methylase are identical to gene products of the broad-host-range plasmid pIP501 from Streptococcus agalactiae. In addition, a 30.5-kb segment is almost identical to pIP501. Genes encoding an aminoglycoside 6-adenylyltransferase, a streptothricin acetyltransferase, and an aminoglycoside phosphotransferase are arranged in tandem on a 7.4-kb fragment as previously reported in Tn5405 from Staphylococcus aureus and in pJH1 from E. faecalis. One interrupted and five complete IS elements as well as three replication genes were also identified. pRE25 was transferred by conjugation to E. faecalis, Listeria innocua, and Lactococcus lactis by means of a transfer region that appears similar to that of pIP501. It is concluded that pRE25 may contribute to the further spread of antibiotic-resistant microorganisms via food into the human community.

Veterinary use and antibiotic resistance.

Globally, an estimated 50% of all antimicrobials serve veterinary purposes. Bacteria that inevitably develop antibiotic resistance in animals comprise food-borne pathogens, opportunistic pathogens and commensal bacteria. The same antibiotic resistance genes and gene transfer mechanisms can be found in the microfloras of animals and humans. Direct contact, food and water link animal and human habitats. The accumulation of resistant bacteria by the use of antibiotics in agriculture and veterinary medicine and the spread of such bacteria via agriculture and direct contamination are documented.

Fluorescent probes for non-invasive bioenergetic studies of whole cyanobacterial cells.

Fluorescent DeltapH and DeltaPsi indicators have been screened for the non-invasive monitoring of bioenergetic processes in whole cells of the cyanobacterium Synechocystis sp. PCC 6803. Acridine yellow and Acridine orange proved to be the best DeltapH indicators for the investigation of thylakoid and cytoplasmic membrane energization: While Acridine yellow indicated only cytosolic energization, Acridine orange showed signals from both the thylakoid lumen and the cytosol that could be separated kinetically. Both indicators were applied successfully to monitor cellular energetics, such as the interplay of linear and cyclic photosynthetic electron transport, osmotic adaptation and solute transport across the cytoplasmic membrane. In contrast, useful membrane potential indicators were more difficult to find, with Di-4-ANEPPS and Brilliant cresyl blue being the only promising candidates for further studies. Finally, Acridine yellow and Acridine orange could also be applied successfully for the thermophilic cyanobacterium Synechococcus elongatus. Different from Synechocystis sp. PCC 6803, where both respiration and ATP hydrolysis could be utilized for cytoplasmic membrane energization, proton extrusion at the cytoplasmic membrane in Synechococcus elongatus was preferentially driven by ATP hydrolysis.

Antimicrobial susceptibility of intestinal bacteria from Swiss poultry flocks before the ban of antimicrobial growth promoters.

From the crop and the caecum of Swiss broilers slaughtered between November 1997 and January 1998, Escherichia coli, enterococci, staphylococci, lactobacilli and Campylobacter species were isolated. After identification to the genus or species level, their minimal inhibitory concentrations (MIC's) for several clinically used antimicrobial agents were determined with the E-Test stripes and compared to those from studies in other European countries. All strains of Enterococcus faecalis (n = 38), E. faecium (27), staphylococci (n = 39) and lactobacilli (n = 14) showed a hundred percent resistance against bacitracin which was included in the feed of the mother animals, but not in the feed of the investigated animals. E.coli strains (n = 60) showed higher resistance incidences than in comparable studies from Finland and Denmark, but lower than those in studies from Italy and Germany. In staphylococci, low resistance rates were observed. A high susceptibility of the 13 Campylobacter jejuni strains was found against therapeutically used antimicrobials. These data can be used as a baseline to determine antibiotic resistance rates after implementation of the growth promotor ban in 1999 in Switzerland.

Characterization of the D-xylulose 5-phosphate/D-fructose 6-phosphate phosphoketolase gene (xfp) from Bifidobacterium lactis.

A D-xylulose 5-phosphate/D-fructose 6-phosphate phosphoketolase (Xfp) from the probiotic Bifidobacterium lactis was purified to homogeneity. The specific activity of the purified enzyme with D-fructose 6-phosphate as a substrate is 4.28 Units per mg of enzyme. K(m) values for D-xylulose 5-phosphate and D-fructose 6-phosphate are 45 and 10 mM, respectively. The native enzyme has a molecular mass of 550,000 Da. The subunit size upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis (90,000 Da) corresponds with the size (92,529 Da) calculated from the amino acid sequence of the isolated gene (named xfp) encoding 825 amino acids. The xfp gene was identified on the chromosome of B. lactis with the help of degenerated nucleotide probes deduced from the common N-terminal amino acid sequence of both the native and denatured enzyme. Comparison of the deduced amino acid sequence of the cloned gene with sequences in public databases revealed high homologies with hypothetical proteins (26 to 55% identity) in 20 microbial genomes. The amino acid sequence derived from the xfp gene contains typical thiamine diphosphate (ThDP) binding sites reported for other ThDP-dependent enzymes. Two truncated putative genes, pta and guaA, were localized adjacent to xfp on the B. lactis chromosome coding for a phosphotransacetylase and a guanosine monophosphate synthetase homologous to products of genes in Mycobacterium tuberculosis. However, xfp is transcribed in B. lactis as a monocistronic operon. It is the first reported and sequenced gene of a phosphoketolase.

Mdt(A), a new efflux protein conferring multiple antibiotic resistance in Lactococcus lactis and Escherichia coli.

The mdt(A) gene, previously designated mef214, from Lactococcus lactis subsp. lactis plasmid pK214 encodes a protein [Mdt(A) (multiple drug transporter)] with 12 putative transmembrane segments (TMS) that contain typical motifs conserved among the efflux proteins of the major facilitator superfamily. However, it also has two C-motifs (conserved in the fifth TMS of the antiporters) and a putative ATP-binding site. Expression of the cloned mdt(A) gene decreased susceptibility to macrolides, lincosamides, streptogramins, and tetracyclines in L. lactis and Escherichia coli, but not in Enterococcus faecalis or in Staphylococcus aureus. Glucose-dependent efflux of erythromycin and tetracycline was demonstrated in L. lactis and in E. coli.

Propionicin SM1, a bacteriocin from Propionibacterium jensenii DF1: isolation and characterization of the protein and its gene.

We purified a bacteriocin from the cell-free supernatant of Propionibacterium jensenii DF1 isolated from Swiss raw milk, and named it propionicin SM1. The heat-stable protein was strongly bactericidal against P. jensenii DSM20274. On the basis of the N-terminal amino acid sequence of the purified protein, a degenerate oligonucleotide probe was designed to locate and clone the corresponding gene of P. jensenii DF1. It hybridized exclusively with the DF1l-resident plasmid pLME106, but not with chromosomal DNA. Sequencing of the 6.9-kb plasmid revealed the targeted amino acid sequence within an open reading frame (ORF4) of 207 amino acids (molecular mass, 22,865 Da). The corresponding gene was named ppnA. It encodes the prepeptide PpnA that is processed to the mature protein (19,942 Da) propionicin SM1. No sequence homology is detectable with known proteins. However, the proposed leader peptide sequence containing 27 amino acids has typical signal peptide features and shows good homology to the leader peptide of Usp45, a protein excreted from Lactococcus lactis (VAN ASSELDONK et al., 1993). Plasmid pLME106 contains at least 9 ORFs, some exhibiting significant homologies to plasmid-encoded functions from other bacteria. The highest identity values were found for ORF1 with the theta replicase (acc. no. U39878) of Brevibacterium linens (58.8%) and ORF6 with the recombinase/invertase (acc. no. AF060871) found in Rhodococcus rhodochrous (46.4%).

Molecular identification of Acetobacter isolates from submerged vinegar production, sequence analysis of plasmid pJK2-1 and application in the development of a cloning vector.

Three new Acetobacter strains were isolated from vinegar. By plasmid profiling they were recognized as genotypically different from each other. Sequencing of the genes for 16S and 23S rRNA and DNA-DNA hybridization of total DNA against DNA of all type strains of Acetobacter identified Acetobacter strains JK2 and V3 as A. europaeus, and Acetobacter strain JK3 as A. intermedius. In contrast to the type strain of A. europaeus (DSM 6160), A. europaeus JK2 and V3 do not require acetic acid for growth and can be successfully transferred between media with and without acetic acid. This phenotypic characteristic enables convenient handling of both strains in genetic studies. Plasmid pJK2-1 from A. europaeus JK2 was used as the basis for shuttle plasmid construction with the aim of developing an efficient vector system for these strains. The entire nucleotide sequence of pJK2-1 was determined. High amino acid identities were found for three open reading frames: Rep (replication protein); Dinjl (DNA damage inducible enzyme); and Dinj2 proteins. A recombinant plasmid pUCJK2-1 (5.6 kb) consisting of the entire plasmid pJK2-1 and the entire plasmid pUC18 was successfully used in transformation experiments. Plasmid pJT2 (5.8 kb) was constructed from pUCJK2-1 with the aim of reactivating the lacZ' gene.

Description of Saccharomyces turicensis sp. nov., a new species from kefyr.

The new species Saccharomyces turicensis sp. nov. isolated from different kefyr grains is described. Although its morphological properties differ, its physiological characteristics come close to those of Saccharomyces bayanus Saccardo and Saccharomyces pastorianus Reess ex E. C. Hansen. However, electrophoretic karyotyping and restriction fragment length polymorphism of the internal transcribed spacer region yield clear differences. Sequences (270 nucleotides) of the D2 domain at the 5'-terminal end of the large subunit ribosomal RNA gene reveal 98.0% identity with Saccharomyces exiguus. Since strains of a particular yeast species usually show less than 1% substitution in the D2 domain, the yeast in question is considered to be a new species. The name Saccharomyces turicensis is proposed indicating the place Zürich (Turicum in Latin) where the yeast had been isolated.

Transcriptional analysis of the rubrerythrin and superoxide dismutase genes of Clostridium perfringens.

We cloned and sequenced a 2.7-kb fragment of chromosomal DNA from Clostridium perfringens containing the superoxide dismutase-encoding gene, sod. Previously, rubrerythrin from C. perfringens had been isolated and its gene (rbr) had been cloned (Y. Lehmann, L. Meile, and M. Teuber, J. Bacteriol. 178:7152-7158, 1996). Northern blot experiments revealed a length of approximately 800 bases for each transcript of rbr and sod of C. perfringens. Thus, rbr and sod each represent a monocistronic operon. Their transcription start points were located by primer extension analyses. sod transcription was shown to depend on the growth phase, and it reached a maximum during the transition from log phase to stationary phase. Neither sod nor rbr transcription was influenced by oxidative stress.

Acquired antibiotic resistance in lactic acid bacteria from food.

Acquired antibiotic resistance, i.e. resistance genes located on conjugative or mobilizable plasmids and transposons can be found in species living in habitats (e.g. human and animal intestines) which are regularly challenged with antibiotics. Most data are available for enterococci and enteric lactobacilli. Raw material from animals (milk and meat) which are inadvertantly contaminated with fecal matters during production will carry antibiotic resistant lactic acid bacteria into the final fermented products such as raw milk cheeses and raw sausages. The discovered conjugative genetic elements of LAB isolated from animals and food are very similar to elements studied previously in pathogenic streptococci and enterococci, e.g. theta-type replicating plasmids of the pAMbeta1, pIP501-family, and transposons of the Tn916-type. Observed resistance genes include known genes like tetM, ermAM, cat, sat and vanA. A composite 29,871 bp resistance plasmid detected in Lactococcus lactis subsp. lactis isolated from a raw milk soft cheese contains tetS previously described in Listeria monocytogenes, cat and str from Staphylococcus aureus. Three out of five IS elements on the plasmid are almost or completely identical to IS1216 present in the vanA resistance transposon Tn1546. These data support the view that in antibiotic challenged habitats lactic acid bacteria like other bacteria participate in the communication systems which transfer resistance traits over species and genus borders. The prevalence of such bacteria with acquired resistances like enterococci is high in animals (and humans) which are regularly treated with antibiotics. The transfer of antibiotic resistant bacteria from animals into fermented and other food can be avoided if the raw substrate milk or meat is pasteurized or heat treated. Antibiotic resistance traits as selectable markers in genetic modification of lactic acid bacteria for different purposes are presently being replaced, e.g. by metabolic traits to generate food-grade vectors.

Spread of antibiotic resistance with food-borne pathogens.

This short review summarizes data on antibiotic resistance profiles of common food-borne pathogens like Salmonella sp., Escherichia coli, Campylobacter sp., Listeria monocytogenes, Clostridium perfringens, Staphylococcus aureus, and coagulase-negative staphylococci. As a flashlight on the literature of the last few years, it provides ample evidence that antibiotic resistance traits have entered the microflora of farm animals and the food produced from them. Molecular analysis of the resistance genes, where available, shows that the food microflora is not separated from its human counterpart and conjugative transfer of resistance genes has been demonstrated in vitro and in a few cases in vivo. For example, for Salmonella typhimurium, resistance towards tetracyclines has increased from zero in 1948 to a 98% level in certain epidemic populations of S. typhimurium DT104 in 1998. The high incidence of food-borne pathogens in raw meat and milk together with a high level of therapeutic, prophylactic and nutritional application of antibiotics in agriculture reveals an antibiotic resistance problem of global dimensions. The resistance problem in human medicine will not be solved if there is a constant influx of resistance genes into the human microflora via the food chain.