Use of metabolomics for the chemotaxonomy of legume-associated Ascochyta and allied genera.

Chemotaxonomy and the comparative analysis of metabolic features of fungi have the potential to provide valuable information relating to ecology and evolution, but have not been fully explored in fungal biology. Here, we investigated the chemical diversity of legume-associated Ascochyta and Phoma species and the possible use of a metabolomics approach using liquid chromatography-mass spectrometry for their classification. The metabolic features of 45 strains including 11 known species isolated from various legumes were extracted, and the datasets were analyzed using chemometrics methods such as principal component and hierarchical clustering analyses. We found a high degree of intra-species consistency in metabolic profiles, but inter-species diversity was high. Molecular phylogenies of the legume-associated Ascochyta/Phoma species were estimated using sequence data from three protein-coding genes and the five major chemical groups that were detected in the hierarchical clustering analysis were mapped to the phylogeny. Clusters based on similarity of metabolic features were largely congruent with the species phylogeny. These results indicated that evolutionarily distinct fungal lineages have diversified their metabolic capacities as they have evolved independently. This whole metabolomics approach may be an effective tool for chemotaxonomy of fungal taxa lacking information on their metabolic content.

Training Intensity Distribution and Changes in Performance and Physiology of a 2nd Place Finisher Team of the Race across America Over a 6 Month Preparation Period.

Aim: To monitor the training intensity distribution (TID) and the development of physiological and performance parameters. Methods: During their preparation period for the RAAM, 4 athletes (plus 1 additional backup racer) performed 3 testing sessions; one before, one after 3, and one after 6 months of training. VO2max, maximal rate of lactate accumulation (dLa/dtmax), critical power, power output at lactate minimum (MLSSP), peak and mean power output during a sprint test, heart rate recovery, isometric strength, jumping height, and body composition were determined. All training sessions were recorded with a power meter. The endurance TID was analyzed based on the time in zone approach, according to a classical 3-zone model, including all power data of training sessions, and a power specific 3-zone model, where time with power output below 50% of MLSSP was not considered. Results: The TID using the classical 3-zone model reflected a pyramidal TID (zone 1: 63 ± 16, zone 2: 28 ± 13 and zone 3: 9 ± 4%). The power specific 3-zone model resulted in a threshold-based TID (zone 1: 48 ± 13, zone 2: 39 ± 10, zone 3: 13 ± 4%). VO2max increased by 7.1 ± 5.3% (P = 0.06). dLa/dtmax decreased by 16.3 ± 8.1% (P = 0.03). Power output at lactate minimum and critical power increased by 10.3 ± 4.1 and 16.8 ± 6.2% (P = 0.01), respectively. No changes were found for strength parameters and jumps. Conclusion: The present study underlines that a threshold oriented TID results in only moderate increases in physiological parameters. The amount of training below 50% of MLSSp (~28% of total training time) is remarkably high. Researchers, trainers, and athletes should pay attention to the different ways of interpreting training power data, to gain realistic insights into the TID and the corresponding improvements in performance and physiological parameters.

Initiation of rivaroxaban in patients with nonvalvular atrial fibrillation at the primary care level: the Swiss Therapy in Atrial Fibrillation for the Regulation of Coagulation (STAR) Study.

BACKGROUND: Rivaroxaban has become an alternative to vitamin-K antagonists (VKA) for stroke prevention in non-valvular atrial fibrillation (AF) patients due to its favourable risk-benefit profile in the restrictive setting of a large randomized trial. However in the primary care setting, physician's motivation to begin with rivaroxaban, treatment satisfaction and the clinical event rate after the initiation of rivaroxaban are not known. METHODS: Prospective data collection by 115 primary care physicians in Switzerland on consecutive nonvalvular AF patients with newly established rivaroxaban anticoagulation with 3-month follow-up. RESULTS: We enrolled 537 patients (73±11years, 57% men) with mean CHADS2 and HAS-BLED-scores of 2.2±1.3 and 2.4±1.1, respectively: 301(56%) were switched from VKA to rivaroxaban (STR-group) and 236(44%) were VKA-naïve (VN-group). Absence of routine coagulation monitoring (68%) and fixed-dose once-daily treatment (58%) were the most frequent criteria for physicians to initiate rivaroxaban. In the STR-group, patient's satisfaction increased from 3.6±1.4 under VKA to 5.5±0.8 points (P<0.001), and overall physician satisfaction from 3.9±1.3 to 5.4±0.9 points (P<0.001) at 3months of rivaroxaban therapy (score from 1 to 6 with higher scores indicating greater satisfaction). In the VN-group, both patient's (5.4±0.9) and physician's satisfaction (5.5±0.7) at follow-up were comparable to the STR-group. During follow-up, 1(0.19%; 95%CI, 0.01-1.03%) ischemic stroke, 2(0.37%; 95%CI, 0.05-1.34%) major non-fatal bleeding and 11(2.05%; 95%CI, 1.03-3.64%) minor bleeding complications occurred. Rivaroxaban was stopped in 30(5.6%) patients, with side effects being the most frequent reason. CONCLUSION: Initiation of rivaroxaban for patients with nonvalvular AF by primary care physicians was associated with a low clinical event rate and with high overall patient's and physician's satisfaction.

Antihypertensive combination therapy in primary care offices: results of a cross-sectional survey in Switzerland.

BACKGROUND: Most hypertensive patients need more than one substance to reach their target blood pressure (BP). Several clinical studies indicate the high efficacy of antihypertensive combinations, and recent guidelines recommend them in some situations even as initial therapies. In general practice they seem widespread, but only limited data are available on their effectiveness under the conditions of everyday life. The objectives of this survey among Swiss primary care physicians treating hypertensive patients were: to know the frequency of application of different treatment modalities (monotherapies, free individual combinations, single-pill combinations); to see whether there are relationships between prescribed treatment modalities and patient characteristics, especially age, treatment duration, and comorbidities; and to determine the response rate (percentage of patients reaching target BP) of different treatment modalities under the conditions of daily practice. METHODS: This cross-sectional, observational survey among 228 randomly chosen Swiss primary care physicians analyzed data for 3,888 consecutive hypertensive patients collected at one single consultation. RESULTS: In this survey, 31.9% of patients received monotherapy, 41.2% two substances, 20.9% three substances, and 4.7% more than three substances. By combination mode, 34.9% took free individual combinations and 30.0% took fixed-dose single-pill combinations. Combinations were more frequently given to older patients with a long history of hypertension and/or comorbidities. In total, 67.8% of patients achieved their BP target according to their physician's judgment. When compared, single-pill combinations were associated with a higher percentage of patients achieving target BP than free individual combinations and monotherapies for the total sample and for patients with comorbidity. CONCLUSION: Antihypertensive combination therapy was widely used in Swiss primary care practices. The number of prescribed substances depended on age, treatment duration, and type and number of comorbidities. Although the response rate was generally modest under the conditions of daily practice, it was higher for single-pill combinations than for monotherapies and free individual combinations. Further studies are needed to confirm these observations.

Measuring accuracy of sphygmomanometers in the medical practices of Swiss primary care physicians.

OBJECTIVE: Arterial hypertension has a high prevalence in most countries. Blood pressure measurements are performed frequently by primary care physicians. Recommendations from different societies emphasise the importance of measuring blood pressure with well maintained and calibrated instruments only. Since appropriate quality control measures are lacking the following survey was conducted in the medical practices of Swiss primary care physicians. METHODS: This is a cross-sectional survey with Swiss primary care physicians. Nine hundred and seventy-five sphygmomanometers used in the daily practice of medicine were compared and calibrated against a certified calibrator. The magnitude of the measuring error before and after calibration was determined. RESULTS: The proportion of the instruments that measured within the required tolerance of ± 3 mmHg over all measuring ranges was 81.4%. The average maintenance time was 5.6 years (± 3.8), and 97% (n = 353) of these instruments had not been maintained for two years (i.e. the recommended maintenance interval) or more. Two years after maintenance the number of devices with measurement errors of more than ± 3 mmHg increased significantly. CONCLUSION: In Swiss primary care practices, the majority of upper arm and wrist sphygmomanometers measured blood pressure within a tolerance of ± 3 mmHg despite low adherence to the recommended maintenance interval. Two years after maintenance the number of sphygmomanometers with measurement errors increased significantly.

Karyotype polymorphism and chromosomal rearrangement in populations of the phytopathogenic fungus, Ascochyta rabiei.

The fungus Ascochyta rabiei is the causal agent of Ascochyta blight of chickpea and the most serious threat to chickpea production. Little is currently known about the genome size or organization of A. rabiei. Given recent genome sequencing efforts, characterization of the genome at a population scale will provide a framework for genome interpretation and direction of future resequencing efforts. Electrophoretic karyotype profiles of 112 isolates from 21 countries revealed 12-16 chromosomes between 0.9 Mb and 4.6 Mb with an estimated genome size of 23 Mb-34 Mb. Three general karyotype profiles A, B, and C were defined by the arrangement of the largest chromosomes. Approximately one-third of isolates (group A) possessed a chromosome larger than 4.0 Mb that was absent from group B and C isolates. The ribosomal RNA gene (rDNA) cluster was assigned to the largest chromosome in all except four isolates (group C) whose rDNA cluster was located on the second largest chromosome (3.2 Mb). Analysis of progeny from an in vitro sexual cross between two group B isolates revealed one of 16 progeny with an rDNA-encoding chromosome larger than 4.0 Mb similar to group A isolates, even though a chromosome of this size was not present in either parent. No expansion of the rDNA cluster was detected in the progeny, indicating the increase in chromosome size was not due to an expansion in number of rDNA repeats. The karyotype of A. rabiei is relatively conserved when compared with published examples of asexual ascomycetes, but labile with the potential for large scale chromosomal rearrangements during meiosis. The results of this study will allow for the targeted sequencing of specific isolates to determine the molecular mechanisms of karyotype variation within this species.

Validation of a multivariate prediction rule for history of periodontitis in a separate population.

AIM: Validation of a previously derived prediction rule for periodontitis in an external population sample. MATERIALS AND METHODS: Age, smoking and self-reported tooth mobility were used in logistic models to predict moderate and severe periodontitis as diagnosed from panoramic radiographs of 246 patients attending private practices in Germany. Coefficients derived from these models were used to predict periodontitis in a representative population-based sample of 3297 residents of the region of Pomerania, Germany. RESULTS: In the full derivation sample, the predictive power of the logistic model as measured by the c-statistic was 0.82 and 0.84 for moderate and severe periodontitis, respectively. In the validation set, these models yielded c-statistics of 0.82 for both moderate and severe periodontitis. Lower c-statistics were obtained among subjects aged 40 years and older in the derivation set (c=0.74 and 0.77), and the performance was poorer in the validation set with c-statistics of 0.69 and 0.72, respectively. CONCLUSIONS: A prediction rule based on age, smoking and self-reported tooth mobility can yield a moderately useful external validity. Validity may be dependent on specific population characteristics, and derivation of a prediction rule based on a clinical subsample of the target population with a larger set of predictors may provide better results in an application.

Differences Between Ascospores and Conidia of Didymella rabiei in Spore Germination and Infection of Chickpea.

ABSTRACT Studies were performed to compare the germination and infection of ascospores and conidia of Didymella rabiei under different temperature and moisture conditions. Germination of ascospores and conidia on cover glasses coated with water agar began after 2 h, with maximum germination (>95%) occurring in 6 h at 20 degrees C. No germination occurred at 0 and 35 degrees C. Ascospores germinated more rapidly than conidia at all temperatures. Germination declined rapidly as the water potential varied from 0 to -4 MPa, although some germination occurred at -6 MPa at 20 and 25 degrees C. Ascospores germinated over a wider range of water potentials than conidia and their germ tubes were longer than those of conidia at most water potentials and temperatures. The optimum temperature for infection and disease development by both ascospores and conidia was around 20 degrees C. Disease severity was higher when ascospores were discharged directly onto plant surfaces from naturally infested chickpea debris compared with aqueous suspensions of ascospores and conidia sprayed onto plants Disease severity increased as the length of the wetness period increased. When dry periods of 6 to 48 h occurred immediately after inoculation, disease severity decreased, except for the shorter periods which had the opposite effect. Disease severity was higher with ascospore inoculum when no dry periods occurred after inoculation.

Cloning of the mating type locus from Ascochyta lentis (teleomorph: Didymella lentis) and development of a multiplex PCR mating assay for Ascochyta species.

The mating type (MAT) locus of the lentil pathogen, Ascochyta lentis, was cloned and characterized using thermal asymmetric interlaced and inverse PCR with primers designed to the HMG-box of Ascochyta rabiei. A multiplex PCR assay for mating type was developed based on MAT idiomorph and flanking sequences. Primers were designed to specifically amplify MAT from several Ascochyta spp. including A. pisi, A. fabae and A. viciae-villosae in addition to A. lentis. Four hundred and fifty and 700 bp fragments were amplified from MAT1-1 and MAT1-2 isolates, respectively, and fragment size correlated perfectly with laboratory crosses using mating type tester strains. MAT-specific PCR allowed rapid scoring of mating type in crude DNA extracts from geographically diverse population samples of A. viciae-villosae from California and Washington State, USA. This co-dominant MAT-specific PCR assay will be a valuable tool for studying the population structure, biology and epidemiology of these fungi.

Sleep deprivation in honey bees.

Rest at night in forager honey bees (Apis mellifera) meets essential criteria of sleep. This paper reports the effect of a 12-h total sleep deprivation (SD) by forced activity on the behaviour of these animals. The behaviour of sleep-deprived animals is compared with that of control animals under LD [periodic alternation between light (L) and darkness (D)] 12 : 12 hours. SD for 12 h during the first D period resulted in a significant difference with respect to the parameter 'hourly amount of antennal immobility' between sleep-deprived and control animals during the remaining L and D periods. This difference did not occur in the L period following the deprivation night, but rather it became obvious at the beginning of the following D period. The increase of the amount of antennal immobility in sleep-deprived bees was accompanied by an increase of the duration of episodes of antennal immobility. Moreover, the latency from 'lights off' to the first episode of antennal immobility lasting 20 s or longer ('deep sleep latency') tended to be shorter in sleep-deprived than in control animals. Disturbing the bees during the day (L period) did not result in such differences between disturbed and control animals. Highest reaction thresholds in sleeping honey bees occur during long episodes of antennal immobility. We therefore conclude that honey bees compensate a sleep deficit by intensification (deepening) of the sleep process and thus that sleep in honey bees, like that in other arthropods and mammals, is controlled by regulatory mechanisms.

Characterization of a New Potyvirus Naturally Infecting Chickpea.

During the 1999 to 2001 growing seasons, symptoms consisting of mosaic, stunting, yellowing, wilting, shortening of internodes, and phloem discoloration were observed in chickpea (Cicer arietinum) grown in the Department of Chuquisaca in southern Bolivia. In some fields, approximately 10% of the plants exhibited viruslike symptoms and suffered greatly reduced seed yields. Lentil (Lens culinaris) was also observed to be infected but not pea (Pisum sativum) or faba bean (Vicia faba) growing in nearby fields. Infected chickpea tissue reacted positively to the potyvirus group-specific monoclonal antibody (MAb), but there was no serological reaction with antisera to the potyviruses Bean yellow mosaic virus, Clover yellow vein virus, Cowpea aphid-borne mosaic virus, Pea seedborne mosaic virus, Bean common mosaic virus, or Bean common mosaic necrosis virus. Western blots of total protein extracts probed with the potyvirus MAb revealed a single band ca. 32 kDa. Comparative sequence analysis of cDNA clones generated from the putative coat protein gene consisted of 282 amino acids (31.9 kDa) and showed moderate identities of 67, 66, 63, 63, and 61% with the coat proteins of potyviruses Pepper severe mosaic virus, Pepper yellow mosaic virus, Potato virus Y, Plum pox virus, and Pepper mottle virus, respectively. Phylogenetic analysis of the coat protein amino acid sequence revealed that this virus is a unique member of the family Potyviridae and is phylogenetically most closely related to a group of Solanaceae-infecting potyviruses rather than to other legumeinfecting potyviruses. The proposed name for the new causal agent is Chickpea yellow mosaic virus.