Comparative study of human erythrocytes by digital holographic microscopy, confocal microscopy, and impedance volume analyzer.

Red blood cell (RBC) parameters such as morphology, volume, refractive index, and hemoglobin content are of great importance for diagnostic purposes. Existing approaches require complicated calibration procedures and robust cell perturbation. As a result, reference values for normal RBC differ depending on the method used. We present a way for measuring parameters of intact individual RBCs by using digital holographic microscopy (DHM), a new interferometric and label-free technique with nanometric axial sensitivity. The results are compared with values achieved by conventional techniques for RBC of the same donor and previously published figures. A DHM equipped with a laser diode (lambda = 663 nm) was used to record holograms in an off-axis geometry. Measurements of both RBC refractive indices and volumes were achieved via monitoring the quantitative phase map of RBC by means of a sequential perfusion of two isotonic solutions with different refractive indices obtained by the use of Nycodenz (decoupling procedure). Volume of RBCs labeled by membrane dye Dil was analyzed by confocal microscopy. The mean cell volume (MCV), red blood cell distribution width (RDW), and mean cell hemoglobin concentration (MCHC) were also measured with an impedance volume analyzer. DHM yielded RBC refractive index n = 1.418 +/- 0.012, volume 83 +/- 14 fl, MCH = 29.9 pg, and MCHC 362 +/- 40 g/l. Erythrocyte MCV, MCH, and MCHC achieved by an impedance volume analyzer were 82 fl, 28.6 pg, and 349 g/l, respectively. Confocal microscopy yielded 91 +/- 17 fl for RBC volume. In conclusion, DHM in combination with a decoupling procedure allows measuring noninvasively volume, refractive index, and hemoglobin content of single-living RBCs with a high accuracy.

Cardenolides, induced responses, and interactions between above- and belowground herbivores of milkweed (Asclepias spp.).

Theory has long predicted allocation patterns for plant defense against herbivory, but only recently have both above- and belowground plant defenses been considered simultaneously. Milkweeds in the genus Asclepias are a classic chemically defended clade of plants with toxic cardenolides (cardiac glycosides) and pressurized latex employed as anti-herbivore weapons. Here we combine a comparative approach to investigate broadscale patterns in allocation to root vs. shoot defenses across species with a species-specific experimental approach to identify the consequences of defense allocational shifts on a specialist herbivore. Our results show phylogenetic conservatism for inducibility of shoot cardenolides by an aboveground herbivore, with only four closely related tropical species showing significant induction; the eight temperate species examined were not inducible. Allocation to root and shoot cardenolides was positively correlated across species, and this relationship was maintained after accounting for phylogenetic nonindependence. In contrast to long-standing theoretical predictions, we found no evidence for a trade-off between constitutive and induced cardenolides; indeed the two were positively correlated across species in both roots and shoots. Finally, specialist root and shoot herbivores of common milkweed (A. syriaca) had opposing effects on latex production, and these effects had consequences for caterpillar growth consistent with latex providing resistance. Although cardenolides were not affected by our treatments, A. syriaca allocated 40% more cardenolides to shoots over roots. We conclude that constitutive and inducible defenses are not trading off across plant species, and shoots of Asclepias are more inducible than roots. Phylogenetic conservatism cannot explain the observed patterns of cardenolide levels across species, but inducibility per se was conserved in a tropical clade. Finally, given that above- and belowground herbivores can systemically alter the defensive phenotype of plants, we concur with recent calls for a whole-plant perspective in testing models of plant defense allocation.

The effect of host social system on parasite population genetic structure: comparative population genetics of two ectoparasitic mites and their bat hosts.

BACKGROUND: The population genetic structure of a parasite, and consequently its ability to adapt to a given host, is strongly linked to its own life history as well as the life history of its host. While the effects of parasite life history on their population genetic structure have received some attention, the effect of host social system has remained largely unstudied. In this study, we investigated the population genetic structure of two closely related parasitic mite species (Spinturnix myoti and Spinturnix bechsteini) with very similar life histories. Their respective hosts, the greater mouse-eared bat (Myotis myotis) and the Bechstein's bat (Myotis bechsteinii) have social systems that differ in several substantial features, such as group size, mating system and dispersal patterns. RESULTS: We found that the two mite species have strongly differing population genetic structures. In S. myoti we found high levels of genetic diversity and very little pairwise differentiation, whereas in S. bechsteini we observed much less diversity, strongly differentiated populations and strong temporal turnover. These differences are likely to be the result of the differences in genetic drift and dispersal opportunities afforded to the two parasites by the different social systems of their hosts. CONCLUSIONS: Our results suggest that host social system can strongly influence parasite population structure. As a result, the evolutionary potential of these two parasites with very similar life histories also differs, thereby affecting the risk and evolutionary pressure exerted by each parasite on its host.

Fecundity of Biomphalaria straminea and B. glabrata in the laboratory: a twelve-month comparative study

In the present comparative study a Biomphalaria straminea sample from Picos (Piauí) showed expressive advantages related to fecundity over a B. glabrata sample from Belo Horizonte (Minas Gerais) such as: higher egg-mass production in 10 out of 12 months of study; higher egg production in all months of study; higher egg per egg-mass ratio in 11 out of 12 months of study; 66% of the egg-masses containing more than 20 eggs while in B. glabrata 70% of the egg-masses showed less than 20 eggs; three times less empty egg capsules than B. glabrata; attainning maximum fecundity in half the time required by B. glabrata. Mortality however was higher and sooner in B. straminea, suggesting higher semelparity in this species than in B. glabrata, a possibility that requires confirmation through long-term studies with other samples of both species. This first finding of a B. straminea sample more fecund than B. glabrata is discussed in relation to other data from the literature, and some recommendations are made on the quantification of fecundity of planorbid snails.

Complementary function and integrated wiring of the evolutionarily distinct Drosophila olfactory subsystems.

To sense myriad environmental odors, animals have evolved multiple, large families of divergent olfactory receptors. How and why distinct receptor repertoires and their associated circuits are functionally and anatomically integrated is essentially unknown. We have addressed these questions through comprehensive comparative analysis of the Drosophila olfactory subsystems that express the ionotropic receptors (IRs) and odorant receptors (ORs). We identify ligands for most IR neuron classes, revealing their specificity for select amines and acids, which complements the broader tuning of ORs for esters and alcohols. IR and OR sensory neurons exhibit glomerular convergence in segregated, although interconnected, zones of the primary olfactory center, but these circuits are extensively interdigitated in higher brain regions. Consistently, behavioral responses to odors arise from an interplay between IR- and OR-dependent pathways. We integrate knowledge on the different phylogenetic and developmental properties of these receptors and circuits to propose models for the functional contributions and evolution of these distinct olfactory subsystems.

Comparative allometry growth of some marine fish digenetic trematodes

Allometric growth variation was compared for Plagioporus idoneus, Lepocreadium pegorchis, Opecoeloides furcatus, Bacciger israelensis, Aphanurus stossichi and Parahurleytrema trachinoti collected from East Mediterranean fishes. The pharynx, the oral and the ventral sucker diameters always showed a negative allometry. The other parameters tested were variable with the species. We study the effects of some environmental factors: the influence of the host species is analysed in Plagioporus idoneous, wich parasitizes Oblada melanura, Diplodus sargus and D. vulgaris and in Lepocreadium pegorchis, wich parasitizes Pagellus erythrinus, Lithognathus mormyrus and Spicara smaris; the influence of the microhabitat and the intensity of infection is analysed in Bacciger israelensis and Aphanurus stossichi, both parasites of Boops boops. We report significant differences with the host species, for the allometric growth of the testes; the effect of the microhabitat was revealed by the hindbody allometric value; no significant difference was detected in relation with the intensity of infection.

Les Partis de défense paysanne dans les pays de l’Europe germanique : étude politologique historique et comparative

Contribution à une meilleure connaissance des partis paysans des pays de l’Europe germanique, exposant sur un mode diachronique leur naissance et leur devenir historique, pour finir précisant, dans une optique comparatiste, quelles sont leurs spécificités communes et les déterminants causaux de leur émergence dans cette partie du continent européen.

Robotic, fluoroscopic or EMG assisted pedicle screw insertion. A CT based comparative study

Introduction: In order to improve safety of pedicle screw placement several techniques have been developed. More recently robotically assisted pedicle insertion has been introduced aiming at increasing accuracy. The aim of this study was to compare this new technique with the two main pedicle insertion techniques in our unit namely fluoroscopically assisted vs EMG aided insertion. Material and methods: A total of 382 screws (78 thoracic,304 lumbar) were introduced in 64 patients (m/f = 1.37, equally distributed between insertion technique groups) by a single experienced spinal surgeon. From those, 64 (10 thoracic, 54 lumbar) were introduced in 11 patients using a miniature robotic device based on pre operative CT images under fluoroscopic control. 142 (4 thoracic, 138 lumbar) screws were introduced using lateral fluoroscopy in 27 patients while 176 (64 thoracic, 112 lumbar) screws in 26 patients were inserted using both fluoroscopy and EMG monitoring. There was no difference in the distribution of scoliotic spines between the 3 groups (n = 13). Screw position was assessed by an independent observer on CTs in axial, sagittal and coronal planes using the Rampersaud A to D classification. Data of lumbar and thoracic screws were processed separately as well as data obtained from axial, sagittal and coronal CT planes. Results: Intra- and interobserver reliability of the Rampersaud classification was moderate, (0.35 and 0.45 respectively) being the least good on axial plane. The total number of misplaced screws (C&D grades) was generally low (12 thoracic and 12 lumbar screws). Misplacement rates were same in straight and scoliotic spines. The only difference in misplacement rates was observed on axial and coronal images in the EMG assisted thoracic screw group with a higher proportion of C or D grades (p <0.05) in that group. Recorded compound muscle action potentials (CMAP) values of the inserted screws were 30.4 mA for the robot and 24.9mA for the freehand technique with a CI of 3.8 of the mean difference of 5.5 mA. Discussion: Robotic placement did improve the placement of thoracic screws but not that of lumbar screws possibly because our misplacement rates in general near that of published navigation series. Robotically assisted spine surgery might therefore enhance the safety of screw placement in particular in training settings were different users at various stages of their learning curve are involved in pedicle instrumentation.

Comparative cost-effectiveness analyses of cardiovascular magnetic resonance and coronary angiography combined with fractional flow reserve for the diagnosis of coronary artery disease.

BACKGROUND: According to recent guidelines, patients with coronary artery disease (CAD) should undergo revascularization if significant myocardial ischemia is present. Both, cardiovascular magnetic resonance (CMR) and fractional flow reserve (FFR) allow for a reliable ischemia assessment and in combination with anatomical information provided by invasive coronary angiography (CXA), such a work-up sets the basis for a decision to revascularize or not. The cost-effectiveness ratio of these two strategies is compared. METHODS: Strategy 1) CMR to assess ischemia followed by CXA in ischemia-positive patients (CMR + CXA), Strategy 2) CXA followed by FFR in angiographically positive stenoses (CXA + FFR). The costs, evaluated from the third party payer perspective in Switzerland, Germany, the United Kingdom (UK), and the United States (US), included public prices of the different outpatient procedures and costs induced by procedural complications and by diagnostic errors. The effectiveness criterion was the correct identification of hemodynamically significant coronary lesion(s) (= significant CAD) complemented by full anatomical information. Test performances were derived from the published literature. Cost-effectiveness ratios for both strategies were compared for hypothetical cohorts with different pretest likelihood of significant CAD. RESULTS: CMR + CXA and CXA + FFR were equally cost-effective at a pretest likelihood of CAD of 62% in Switzerland, 65% in Germany, 83% in the UK, and 82% in the US with costs of CHF 5'794, euro 1'517, £ 2'680, and $ 2'179 per patient correctly diagnosed. Below these thresholds, CMR + CXA showed lower costs per patient correctly diagnosed than CXA + FFR. CONCLUSIONS: The CMR + CXA strategy is more cost-effective than CXA + FFR below a CAD prevalence of 62%, 65%, 83%, and 82% for the Swiss, the German, the UK, and the US health care systems, respectively. These findings may help to optimize resource utilization in the diagnosis of CAD.

Energy expenditure by doubly labeled water: validation in humans and proposed calculation.

To further validate the doubly labeled water method for measurement of CO2 production and energy expenditure in humans, we compared it with near-continuous respiratory gas exchange in nine healthy young adult males. Subjects were housed in a respiratory chamber for 4 days. Each received 2H2(18)O at either a low (n = 6) or a moderate (n = 3) isotope dose. Low and moderate doses produced initial 2H enrichments of 5 and 10 X 10(-3) atom percent excess, respectively, and initial 18O enrichments of 2 and 2.5 X 10(-2) atom percent excess, respectively. Total body water was calculated from isotope dilution in saliva collected at 4 and 5 h after the dose. CO2 production was calculated by the two-point method using the isotopic enrichments of urines collected just before each subject entered and left the chamber. Isotope enrichments relative to predose samples were measured by isotope ratio mass spectrometry. At low isotope dose, doubly labeled water overestimated average daily energy expenditure by 8 +/- 9% (SD) (range -7 to 22%). At moderate dose the difference was reduced to +4 +/- 5% (range 0-9%). The isotope elimination curves for 2H and 18O from serial urines collected from one of the subjects showed expected diurnal variations but were otherwise quite smooth. The overestimate may be due to approximations in the corrections for isotope fractionation and isotope dilution. An alternative approach to the corrections is presented that reduces the overestimate to 1%.