Wolbachia, Cardinium and climate: an analysis of global data.

Bacterial endosymbionts are very common in terrestrial arthropods, but infection levels vary widely among populations. Experiments and within-species comparisons suggest that environmental temperature might be important in explaining this variation. To investigate the importance of temperature, at broad geographical and taxonomic scales, we extended a global database of terrestrial arthropods screened for Wolbachia and Cardinium. Our final dataset contained data from more than 117 000 arthropods (over 2500 species) screened for Wolbachia and more than 18 000 arthropods (over 800 species) screened for Cardinium, including samples from 137 different countries, with mean temperatures varying from -6.5 to 29.2°C. In insects and relatives, Cardinium infection showed a clear and consistent tendency to increase with temperature. For Wolbachia, a tendency to increase with temperature in temperate climates is counteracted by reduced prevalence in the tropics, resulting in a weak negative trend overall. We discuss the implications of these results for natural and introduced symbionts in regions affected by climate change.

An emerging zoonotic clone in the Netherlands provides clues to virulence and zoonotic potential of Streptococcus suis.

Streptococcus suis is a zoonotic swine pathogen and a major public health concern in Asia, where it emerged as an important cause of bacterial meningitis in adults. While associated with food-borne transmission in Asia, zoonotic S. suis infections are mainly occupational hazards elsewhere. To identify genomic differences that can explain zoonotic potential, we compared whole genomes of 98 S. suis isolates from human patients and pigs with invasive disease in the Netherlands, and validated our observations with 18 complete and publicly available sequences. Zoonotic isolates have smaller genomes than non-zoonotic isolates, but contain more virulence factors. We identified a zoonotic S. suis clone that diverged from a non-zoonotic clone by means of gene loss, a capsule switch, and acquisition of a two-component signalling system in the late 19th century, when foreign pig breeds were introduced. Our results indicate that zoonotic potential of S. suis results from gene loss, recombination and horizontal gene transfer events.

Effects of 5 days of head-down bed rest, with and without short-arm centrifugation as countermeasure, on cardiac function in males (BR-AG1 study).

This study examined cardiac remodeling and functional changes induced by 5 days of head-down (-6°) bed rest (HDBR) and the effectiveness of short-arm centrifugation (SAC) in preventing them in males. Twelve healthy men (mean age: 33 ± 7) were enrolled in a crossover design study (BR-AG1, European Space Agency), including one sedentary (CTRL) and two daily SAC countermeasures (SAC1, 30 min continuously; SAC2, 30 min intermittently) groups. Measurements included plasma and blood volume and left ventricular (LV) and atrial (LA) dimensions by transthoracic echocardiography (2- and 3-dimensional) and Doppler inflows. Results showed that 5 days of HDBR had a major impact on both the geometry and cardiac function in males. LV mass and volume decreased by 16 and 14%, respectively; LA volume was reduced by 36%; Doppler flow and tissue Doppler velocities were reduced during early filling by 18 and 12%, respectively; and aortic flow velocity time integral was decreased by 18% with a 3% shortening of LV ejection time. These modifications were presumably due to decreased physiological loading and dehydration, resulting in reduced plasma and blood volume. All these changes were fully reversed 3 days after termination of HDBR. Moreover, SAC was not able to counteract these changes, either when applied continuously or intermittently.

Comparative population genomics in animals uncovers the determinants of genetic diversity.

Genetic diversity is the amount of variation observed between DNA sequences from distinct individuals of a given species. This pivotal concept of population genetics has implications for species health, domestication, management and conservation. Levels of genetic diversity seem to vary greatly in natural populations and species, but the determinants of this variation, and particularly the relative influences of species biology and ecology versus population history, are still largely mysterious. Here we show that the diversity of a species is predictable, and is determined in the first place by its ecological strategy. We investigated the genome-wide diversity of 76 non-model animal species by sequencing the transcriptome of two to ten individuals in each species. The distribution of genetic diversity between species revealed no detectable influence of geographic range or invasive status but was accurately predicted by key species traits related to parental investment: long-lived or low-fecundity species with brooding ability were genetically less diverse than short-lived or highly fecund ones. Our analysis demonstrates the influence of long-term life-history strategies on species response to short-term environmental perturbations, a result with immediate implications for conservation policies.

Population genomics of eusocial insects: the costs of a vertebrate-like effective population size.

The evolution of reproductive division of labour and social life in social insects has lead to the emergence of several life-history traits and adaptations typical of larger organisms: social insect colonies can reach masses of several kilograms, they start reproducing only when they are several years old, and can live for decades. These features and the monopolization of reproduction by only one or few individuals in a colony should affect molecular evolution by reducing the effective population size. We tested this prediction by analysing genome-wide patterns of coding sequence polymorphism and divergence in eusocial vs. noneusocial insects based on newly generated RNA-seq data. We report very low amounts of genetic polymorphism and an elevated ratio of nonsynonymous to synonymous changes ­ a marker of the effective population size ­ in four distinct species of eusocial insects, which were more similar to vertebrates than to solitary insects regarding molecular evolutionary processes. Moreover, the ratio of nonsynonymous to synonymous substitutions was positively correlated with the level of social complexity across ant species. These results are fully consistent with the hypothesis of a reduced effective population size and an increased genetic load in eusocial insects, indicating that the evolution of social life has important consequences at both the genomic and population levels.

Reference-free transcriptome assembly in non-model animals from next-generation sequencing data.

Next-generation sequencing (NGS) technologies offer the opportunity for population genomic study of non-model organisms sampled in the wild. The transcriptome is a convenient and popular target for such purposes. However, designing genetic markers from NGS transcriptome data requires assembling gene-coding sequences out of short reads. This is a complex task owing to gene duplications, genetic polymorphism, alternative splicing and transcription noise. Typical assembling programmes return thousands of predicted contigs, whose connection to the species true gene content is unclear, and from which SNP definition is uneasy. Here, the transcriptomes of five diverse non-model animal species (hare, turtle, ant, oyster and tunicate) were assembled from newly generated 454 and Illumina sequence reads. In two species for which a reference genome is available, a new procedure was introduced to annotate each predicted contig as either a full-length cDNA, fragment, chimera, allele, paralogue, genomic sequence or other, based on the number of, and overlap between, blast hits to the appropriate reference. Analyses showed that (i) the highest quality assemblies are obtained when 454 and Illumina data are combined, (ii) typical de novo assemblies include a majority of irrelevant cDNA predictions and (iii) assemblies can be appropriately cleaned by filtering contigs based on length and coverage. We conclude that robust, reference-free assembly of thousands of genes from transcriptomic NGS data is possible, opening promising perspectives for transcriptome-based population genomics in animals. A Galaxy pipeline implementing our best-performing assembling strategy is provided.

The Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation is less accurate in patients with Type 2 diabetes when compared with healthy individuals.

AIMS: To analyse the performances of Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) and of Modification of Diet in Renal Disease (MDRD) study equations to estimate glomerular filtration rate (GFR) in patients with Type 2 diabetes mellitus with GFRs >60 ml/min and in healthy volunteers. METHODS: This cross-sectional study included 111 individuals (56 patients with Type 2 diabetes and 55 healthy volunteers), aged 58 ± 9 years; 54 individuals were men (49%) and ninety-eight (88%) were white. Glomerular filtration rate was measured by the (51) Cr-EDTA single-injection method ((51) Cr-GFR) and estimated according to the standardized MDRD and CKD-EPI equations. Serum creatinine was measured by a traceable Jaffe method. Bland-Altman analysis was used to examine the agreement between measured and estimated GFR. Bias, accuracy and precision were evaluated. RESULTS: In diabetic individuals, (51) Cr-GFR was 106 ± 27 ml/min/1.73 m(2) , CKD-EPI-estimated GFR 82 ± 18 ml/min/1.73 m(2) and MDRD-estimated GFR 80 ± 21 ml/min/1.73 m(2) (P < 0.001). In healthy volunteers, the corresponding values were 98 ± 20, 89 ± 13 and 84 ± 14 ml/min/1.73 m(2) (P < 0.001). The accuracy of CKD-EPI (P30) was higher in healthy volunteers than in diabetic patients (90 vs. 66%, respectively, P < 0.001). The MDRD equation performed as poorly as the CKD-EPI equation in individuals with Type 2 diabetes. CONCLUSIONS: The CKD-EPI equation is less accurate in patients with Type 2 diabetes when compared with healthy individuals, with a 2.5-fold greater bias.

The role of echocardiography in the assessment of cardiac function in weightlessness-Our experience during parabolic flights.

Parabolic flight (PF) elicits changes in hydrostatic pressure gradients, resulting in increase (at 0Gz) or decrease (at 1.8Gz) in cardiac preload. The magnitude of these changes on left ventricular (LV) and atrial (LA) volumes, as well as on myocardial velocities, strain and strain rates, is largely unknown. Using real-time 3D (RT3DE) and Doppler tissue echocardiographic imaging (DTI) during PF in normal subjects in standing position, we showed that both LV and LA volumes were decreased at 1.8Gz and increased at 0Gz by about 20% and 40%, respectively. Previous 2D or M-mode studies underestimated such changes. Also, preload dependence was confirmed for systolic and diastolic velocities, and peak systolic strain, while strain rates were preload independent, probably reflecting intrinsic myocardial properties. Low body negative pressure at -50mmHg applied during 0Gz was effective in restoring 1Gz levels. RT3DE and DTI during PF are feasible, allowing the evaluation of the cardiac function under different loading conditions.

Evaluation of alterations on mitral annulus velocities, strain, and strain rates due to abrupt changes in preload elicited by parabolic flight.

We tested the hypothesis that in normal subjects, cardiac tissue velocities, strain, and strain rates (SR), measured by Doppler tissue echocardiography (DTE), are preload dependent. To accomplish it, immediately preceding image acquisition, reversible, repeatable, acute nonpharmacological changes in preload were induced by parabolic flight. DTE has been proposed as a new approach to assess left ventricular regional myocardial function by computing tissue velocities, strain, and SR. However, preload dependence of these parameters in normal subjects still remains controversial. DTE images (Philips) were obtained in 10 normal subjects in standing upright position at normogravity (1 Gz), hypergravity (1.8 Gz), and microgravity (0 Gz) with and without -50 mmHg lower body negative pressure (LBNP). Myocardial velocity curves in the basal interventricular septum were reconstituted offline from DTE images, from which peak systolic (S'), early (E') and late (A') diastolic velocities, SR, and peak systolic strain (PSepsilon) were measured and averaged over four beats. At 1.8 Gz (reduced venous return), S', E', and A' decreased by 21%, 21%, and 26%, respectively, compared with 1-Gz values, while at 0 Gz (augmented venous return), E', A', and PSepsilon increased by 57%, 53%, and 49%, respectively. LBNP reduced E' and PSepsilon. In conclusion, our results were in agreement with those obtained in animal models, in which preload was changed in a controlled, acute, and reversible manner, and image acquisition was performed immediately following preload modifications. The hypothesis of preload dependence was confirmed for S', E', A', and PSepsilon, while SR appeared to be preload independent, probably reflecting intrinsic myocardial properties.

Objective evaluation of changes in left ventricular and atrial volumes during parabolic flight using real-time three-dimensional echocardiography.

We tested the feasibility of real-time three-dimensional (3D) echocardiographic (RT3DE) imaging to measure left heart volumes at different gravity during parabolic flight and studied the effects of lower body negative pressure (LBNP) as a countermeasure. Weightlessness-related changes in cardiac function have been previously studied during spaceflights using both 2D and 3D echocardiography. Several technical factors, such as inability to provide real-time analysis and the need for laborious endocardial definition, have limited its usefulness. RT3DE imaging overcomes these limitations by acquiring real-time pyramidal data sets encompassing the entire ventricle. RT3DE data sets were obtained (Philips 7500, X3) during breath hold in 16 unmedicated normal subjects in upright standing position at different gravity phases during parabolic flight (normogravity, 1 Gz; hypergravity, 1.8 Gz; microgravity, 0 Gz), with LBNP applied (-50 mmHg) at 0 Gz in selected parabolas. RT3DE imaging during parabolic flight was feasible in 14 of 16 subjects. Data were analyzed (Tomtec) to quantify left ventricular (LV) and atrial (LA) volumes at end diastole and end systole, which significantly decreased at 1.8 Gz and increased at 0 Gz. While ejection fraction did not change with gravity, stroke volume was reduced by 16% at 1.8 Gz and increased by 20% at 0 Gz, but it was not significantly different from 1 Gz values with LBNP. RT3DE during parabolic flight is feasible and provides the basis for accurate quantification of LV and LA volume changes with gravity. As LBNP counteracted the increase of LV and LA volumes caused by changes in venous return, it may be effectively used for preventing cardiac dilatation during 0 Gz.

The efficacy of folk medicines in the management of type 2 diabetes mellitus: results of a randomized controlled trial of Syzygium cumini (L.) Skeels.

OBJECTIVE: To investigate whether a tea prepared from leaves of jambolan, Syzygium cumini (L.) Skeels, has an antihyperglycaemic effect in patients with type 2 diabetes mellitus. RESEARCH DESIGN AND METHODS: Patients with type 2 diabetes mellitus were enrolled in a double-blind, double-dummy, randomized clinical trial. The three experimental groups received a tea prepared from leaves of S. cumini plus placebo tablets, placebo tea plus glyburide tablets or placebo tea plus placebo tablets. RESULTS: In total, 27 patients were allocated to one of the treatment groups and followed for 28 days. Fasting blood glucose levels decreased significantly with glyburide and did not change with S. cumini tea or placebo. Body mass index, creatinine, gamma-glutamyl transferase, alkaline phosphatase, aspartate aminotransferase (SGOT), alanine aminotransferase (SGPT), 24-h glicosuria, 24-h proteinuria, triglycerides, total, low-density lipoprotein and high-density lipoprotein cholesterol did not vary significantly between the different groups. CONCLUSIONS: Tea prepared from leaves of S. cumini has no hypoglycaemic effect.

Improved quantification of left ventricular mass based on endocardial and epicardial surface detection with real time three dimensional echocardiography.

OBJECTIVE: To develop a technique for volumetric analysis of real time three dimensional echocardiography (RT3DE) data aimed at quantifying left ventricular (LV) mass and to validate the technique against magnetic resonance (MR) assumed as the reference standard. DESIGN: RT3DE, which has recently become widely available, provides dynamic pyramidal data structures that encompass the entire heart and allows four dimensional assessment of cardiac anatomy and function. However, analysis techniques for the quantification of LV mass from RT3DE data are fundamentally two dimensional, rely on geometric modelling, and do not fully exploit the volumetric information contained in RT3DE datasets. Twenty one patients underwent two dimensional echocardiography (2DE), RT3DE, and cardiac MR. LV mass was measured from 2DE and MR images by conventional techniques. RT3DE data were analysed to semiautomatically detect endocardial and epicardial LV surfaces by the level set approach. From the detected surfaces, LV mass was computed directly in the three dimensional space as voxel counts. RESULTS: RT3DE measurement was feasible in 19 of 21 patients and resulted in higher correlation with MR (r = 0.96) than did 2DE (r = 0.79). RT3DE measurements also had a significantly smaller bias (-2.1 g) and tighter limits of agreement (2SD = +/-23 g) with MR than did the 2DE values (bias (2SD) -34.9 (50) g). Additionally, interobserver variability of RT3DE (12.5%) was significantly lower than that of 2DE (24.1%). CONCLUSIONS: Direct three dimensional model independent LV mass measurement from RT3DE images is feasible in the clinical setting and provides fast and accurate assessment of LV mass, superior to the two dimensional analysis techniques.

Feasibility of real-time 3D echocardiography in weightlessness during parabolic flight.

Aim of the study was to test the feasibility of transthoracic real-time 3D (Philips) echocardiography (RT3D) during parabolic flight, to allow direct measurement of heart chambers volumes modifications during the parabola. One RT3D dataset corresponding to one cardiac cycle was acquired at each gravity phase (1 Gz, 1.8 Gz, 0 Gz, 1.8 Gz) during breath-hold in 8 unmedicated normal subjects (41 +/- 8 years old) in standing upright position. Preliminary results, obtained by semi-automatically tracing left ventricular (LV) and left atrial (LA) endocardial contours in multiple views (Tomtec), showed a significant (p<0.05) reduction, compared to 1 Gz, of LV and LA volumes with 1.8 Gz, and a significant increase with 0 Gz. Further analysis will focus on the right heart.

Effects of aging on left atrial reservoir, conduit, and booster pump function: a multi-institution acoustic quantification study.

OBJECTIVE: To assess the feasibility of measuring left atrial (LA) function with acoustic quantification (AQ) and then assess the effects of age and sex on LA reservoir, conduit, and booster pump function. PATIENTS AND SETTING: 165 subjects without cardiovascular disease, 3-79 years old, were enrolled by six tertiary hospital centres. INTERVENTIONS: Continuous LA AQ area data were acquired and signal averaged to form composite waveforms which were analysed off-line. MAIN OUTCOME MEASURES: Parameters of LA performance according to age and sex. RESULTS: Signal averaged LA waveforms were sufficiently stable and detailed to allow automated analysis in all cases. An age related increase in LA area was noted. LA reservoir function did not vary with age or sex. All parameters of LA passive and active emptying revealed a significant age dependency. Overall, the passive emptying phase accounted for 66% of total LA emptying ranging from 76% in the youngest to 44% in the oldest decade. LA contraction accounted for 34% of atrial emptying in all subjects combined with the older subjects being more dependent on atrial booster pump function. When adjusted for atrial size, there were no sex related differences in LA function. CONCLUSIONS: LA reservoir, conduit, and booster pump function can be assessed with automated analysis of signal averaged LA area waveforms. As LA performance varies with age, establishment of normal values should enhance the evaluation of pathologic states in which LA function is important.

Electronic transmission of digital echocardiographic studies: effects of MPEG compression.

The acquisition, storage and retrieval of digital echocardiographic studies greatly facilitates image review and quantitation and permits the transmission of studies electronically. However, the considerable size of digital echocardiographic data files makes transmission over existing networks slow and impractical. Reduction in the size of these data files can be accomplished with digital image compression. We sought to evaluate the effects of MPEG-1 compression on the transfer time of digital echocardiographic studies over currently available network connections. Ninety consecutive routine clinical echocardiographic studies were randomly compressed at one of three compression ratios 60:1, 80:1, or 120:1 and sent to a receiving terminal using simulated transmission rates. Compression of digital echocardiographic studies at these ratios which, have been shown to maintain diagnostic image quality, reduced the size of digital echocardiographic studies to less than 1% of their original sizes which allowed transmission of echocardiographic studies over networks using 3ISDN or T1 lines with minimal waiting time.

Normal values of regional left ventricular endocardial motion: multicenter color kinesis study.

Our goal was to establish normal values for quantitative color kinesis indexes of left ventricular (LV) wall motion over a wide range of ages, which are required for objective diagnosis of regional systolic and diastolic dysfunction. Color-encoded images were obtained in 194 normal subjects (95 males, 99 females, age 2 mo to 79 yr) in four standard views. Quantitative indexes of magnitude and timing of systolic and diastolic function were studied for age- and gender-related differences. Normal limits of all ejection and filling indexes were in a narrow range (< or =25% of the mean), with no major gender-related differences. Despite invariable ejection fractions, both peak filling and ejection rates decreased with age (30 and 20%, correspondingly) with a concomitant increase in mean filling and ejection times, resulting in five- and twofold increases in the late to early filling and ejection ratios, correspondingly. Diastolic asynchrony increased with age (from 4.7 +/- 2.0 to 6.4 +/- 3.2 from the 2nd to 7th decade). The normal values of color kinesis indexes should allow objective detection of regional LV systolic and diastolic dysfunction.

Power Doppler imaging as a basis for automated endocardial border detection during left ventricular contrast enhancement.

Echocardiographic evaluation of left ventricular (LV) systolic function relies on endocardial visualization, which can be improved when necessary using contrast enhancement. However, there is no method to automatically detect the endocardial boundary from contrast-enhanced images. We hypothesized that this could be achieved using harmonic power Doppler imaging. Twenty-two patients were studied in two protocols: (1) 11 patients with poorly visualized endocardium (> 3 contiguous segments not visualized) and (2) 11 consecutive patients referred for dobutamine stress echocardiography who were studied at rest and at peak dobutamine infusion. Patients were imaged in the apical four-chamber view using harmonic power Doppler mode (HP SONOS 5500) during LV contrast enhancement (Optison or Definity DMP115). Digital images were analyzed using custom software designed to automatically extract the endocardial boundary from power Doppler color overlays. LV cavity area was automatically measured frame-by-frame throughout the cardiac cycle, and fractional area change calculated and compared with those obtained by manually tracing the endocardial boundary in end-systolic and end-diastolic gray scale images. Successful border detection and tracking throughout the cardiac cycle was possible in 9 of 11 patients with poor endocardial definition and in 10 of 11 unselected patients undergoing dobutamine stress testing. Fractional area change obtained from power Doppler images correlated well with manually traced area changes (r = 0.82 and r = 0.97, in protocols 1 and 2, respectively). Harmonic power Doppler imaging with contrast may provide a simple method for semi-automated border detection and thus facilitate the objective evaluation of LV function both at rest and under conditions of stress testing. This methodology may prove to be particularly useful in patients with poorly visualized endocardium.

The role of echocardiographic harmonic imaging and contrast enhancement for improvement of endocardial border delineation.

Despite advances in imaging technology, many myocardial segments remain poorly visualized with echocardiography; however, both contrast enhancement and harmonic imaging have shown promise for improving endocardial definition. Fifty subjects with technically limited echocardiograms were studied with fundamental and harmonic imaging as well as during echocardiographic contrast injection. Overall endocardial visualization scores improved with both techniques compared with fundamental imaging. Harmonic imaging improved endocardial visualization in 43% of all segments and in 57% of segments nonvisualized with fundamental imaging. The benefit of harmonic imaging was seen in all segments. Contrast echocardiography had similar overall improvements in visualization (42% of all segments, 67% of segments nonvisualized with fundamental imaging) but was not helpful in all regions. Harmonic imaging outperformed contrast in 9 of 22 segments, whereas contrast was superior in 4 of 22. In a subgroup of patients with very poor images, contrast enhancement was superior, with a greater increase in overall score and a higher salvage rate than harmonic (68% vs 40%).