Echocardiographic chamber quantification in a healthy Dutch population

Aim

For accurate interpretation of echocardiographic measurements normative data are required, which are provided by guidelines. For this article, the hypothesis was that these cannot be extrapolated to the Dutch population, since in Dutch clinical practice often higher values are found, which may not be pathological but physiological. Therefore this study aimed to 1) obtain and propose normative values for cardiac chamber quantification in a healthy Dutch population and 2) determine influences of baseline characteristics on these measurements.

Methods

Prospectively recruited healthy subjects, aged 20–72 years (at least 28 subjects per age decade, equally distributed for gender) underwent physical examination and 2D and 3D echocardiography. Both ventricles and atria were assessed and volumes were calculated.

Results

147 subjects were included (age 44 ± 14 years, 50% female). Overall, feasibility was good for both linear and volumetric measurements. Linear and volumetric parameters were consistently higher than current guidelines recommend, while functional parameters were in line with the guidelines. This was more so in the older population. 3D volumes were higher than 2D volumes. Gender dependency was seen in all body surface area (BSA) corrected volumes and with increasing age, ejection fractions decreased.

Conclusion

This study provides 2D and 3D echocardiographic reference ranges for both ventricles and atria derived from a healthy Dutch population. BSA indexed volumes are gender-dependent, age did not influence ventricular volumes and a rise in blood pressure was independently associated with increased right ventricular volumes. The higher volumes found may be indicative for the Dutch population being the tallest in the world.

     

μ-Slide Chemotaxis: A new chamber for long-term chemotaxis studies

Background

Effective tools for measurement of chemotaxis are desirable since cell migration towards given stimuli plays a crucial role in tumour metastasis, angiogenesis, inflammation, and wound healing. As for now, the Boyden chamber assay is the longstanding "gold-standard" for in vitro chemotaxis measurements. However, support for live cell microscopy is weak, concentration gradients are rather steep and poorly defined, and chemotaxis cannot be distinguished from migration in a single experiment.

Results

Here, we describe a novel all-in-one chamber system for long-term analysis of chemotaxis in vitro that improves upon many of the shortcomings of the Boyden chamber assay. This chemotaxis chamber was developed to provide high quality microscopy, linear concentration gradients, support for long-term assays, and observation of slowly migrating cells via video microscopy. AlexaFluor 488 dye was used to demonstrate the establishment, shape and time development of linear chemical gradients. Human fibrosarcoma cell line HT1080 and freshly isolated human umbilical vein endothelial cells (HUVEC) were used to assess chemotaxis towards 10% fetal calf serum (FCS) and FaDu cells' supernatant. Time-lapse video microscopy was conducted for 48 hours, and cell tracking and analysis was performed using ImageJ plugins. The results disclosed a linear steady-state gradient that was reached after approximately 8 hours and remained stable for at least 48 hours. Both cell types were chemotactically active and cell movement as well as cell-to-cell interaction was assessable.

Conclusions

Compared to the Boyden chamber assay, this innovative system allows for the generation of a stable gradient for a much longer time period as well as for the tracking of cell locomotion along this gradient and over long distances. Finally, random migration can be distinguished from primed and directed migration along chemotactic gradients in the same experiment, a feature, which can be qualified via cell morphology imaging.     

Design of a resilient ring for middle ear’s chamber stapes prosthesis

Abstract

This paper presents the process of designing a new elastic element replacing a membrane in the chamber stapes prosthesis (ChSP). The results of the study are volume displacement characteristics obtained for the prosthesis and physiological stapes. Simulation tests on a 3D CAD model have confirmed that a properly designed ring can stimulate perilymph with the same or greater efficacy as the physiological stapes footplate placed on the elastic annular ligament. The ChSP with a new elastic element creates a good chance of improving hearing in patients suffering from otosclerosis.     

In situ fluorochrome calcein marking of deep-sea molluscs using a new growth chamber

A new instrument designed for in situ chemical marking experiments was developed and applied to the deep-sea seep clams Calyptogena soyoae and Calyptogena okutanii. Fluorochrome calcein was used for vital staining of four living clams kept in a specially designed in situ growth chamber that was placed on the seafloor at a cold-seep site off Hatsushima Island, Sagami Bay, central Japan. The shell margins of both C. soyoae and C. okutanii were clearly stained, forming a thin fluorescent band. This method will be useful for age and growth-rate determinations of many other marine organisms with accretionary growing skeletons.     

Dosimetry evaluation of a cylindrical waveguide chamber for unrestrained small rodents at 1.9 GHz

An exposure system, consisting of four identical cylindrical waveguide chambers, was developed for studying the effects of radiofrequency (RF) energy on laboratory mice at a frequency of 1.9 GHz. The chamber was characterized for RF dose rate as a function of animal body mass and dose rate variations due to animal movement in the cage. Dose rates were quantified in terms of whole‐body average (WBA) specific absorption rate (SAR), brain average (BA) SAR and peak spatial‐average (PSA) SAR using measurement and computational methods. Measurements were conducted on mouse cadavers in a multitude of possible postures and positions to evaluate the variations of WBA‐SAR and its upper and lower bounds, while computations utilizing the finite‐difference time‐domain method together with a heterogeneous mouse model were performed to determine variations in BA‐SAR and the ratio of PSA‐SAR to WBA‐SAR. Measured WBA‐SAR variations were found to be within the ranges of 9–23.5 W/kg and 5.2–13.8 W/kg per 1 W incident power for 20 and 40 g mice, respectively. Computed BA‐SAR variations were within the ranges of 3.2–10.1 W/kg and 3.3–9.2 W/kg per 1 W incident power for 25 and 30 g mouse models, respectively. Ratios of PSA‐SAR to WBA‐SAR, averaged over 0.5 mg and 5 mg tissue volumes, were observed to be within the ranges of 6–15 and 4–10, respectively. Bioelectromagnetics 33:575–584, 2012. © 2012 Wiley Periodicals, Inc.     

Evaluation of dose equivalent using a tissue-equivalent ionization chamber and a Geiger-Müller dosimeter

Summary When detailed neutron energy spectrum data are lacking for a mixed field of neutrons and photons, it is permissible when estimating the dose equivalent to assume that the quality factor for the neutrons is 10. With this assumption, it is shown that the responses of a tissue-equivalent ionization chamber and a Geiger-Müller dosimeter can be used to obtain an acceptable approximation of the dose equivalent in the mixed field without requiring precise knowledge of the relative neutron sensitivity of the Geiger-Müller dosimeter.This investigation was supported by Contract EP-78-S-02-4733 from the Department of Energy to the Radiological Research Laboratory/Department of Radiology and by Grant No. CA13696 to the Cancer Center/Institute of Cancer Research, awarded by National Cancer Institute, DHEW     

Should all patients receive dual chamber pacing ICDs? The rationale for the DAVID trial

All of the prospective multicenter trials that support the use of implantable defibrillators have used single chamber pacemakers/implantable cardiovertor defibrillators (ICDs). Despite the significantly increased cost of dual chamber pacemaker/ICD devices and the lack of outcome data, these devices accounted for approximately two-thirds of the ICDs implanted in the United States during the 12 months ending April 2001. Dual chamber pacemaker trials have not provided data that would support this trend, but the high incidence of atrial fibrillation, bradycardia, and congestive heart failure, as comorbid conditions, suggest that the situation could be different in the defibrillator patient population. The DAVID (Dual Chamber and VVI Implantable Defibrillator) trial is designed to measure the incremental benefit of dual chamber pacemaker/ICDs.     

A mechanical interpretation of pressure chamber measurements--what does the strength of the squeeze tell us?

Argument still continues about what properties of a plant organ the pressure chamber measures. A mechanical (as opposed to a thermodynamic) analysis is made of the system squeezed by the pressurized gas, the non-gaseous part of the leaf. The boundary of the system is defined so that it remains at constant mass, and constant density is assumed, during the squeeze. This is equivalent to assuming constant volume. On those assumptions, it is shown that the liquid is brought to the cut surface by a change of shape of the system. Generic mechanical principles are then used to deduce a priori, a quantitative interpretation of the balance pressure. The formal mechanical interpretation involves two variables, the interfacial tension and the change in surface area, which cannot currently be measured. Instead of these, we used two related variables which can be measured, the mass fraction of water in the leaf (Q) and the maximum mass fraction of water at full saturation (Qx) to deduce an approximate mechanical interpretation. When Q is close to Qx, we deduced that the balance pressure (Pb) required for the shape change should be approximately proportional to the reduction in mass in changing from Qx to Q, a variable called the relative water loss (RWL). The constant of proportionality (kappa) is a basic characteristic of the type of leaf used, and the final relation, Pb=kappa (RWL) is called Relation A. We then deduce that the constant kappa should be an approximately linear function of Qx. The linear function is defined by limiting values, so that when Qx is 1, kappa is predicted to be 0 bar, and at the other extreme, when Qx is 0, kappa is predicted to be in the range 500-1000 bar. This is called Relation B. Experiments with 32 leaves from 10 species are used to test the mechanical interpretation. The results showed that Relation A was a reasonable approximation for most of the tested leaves. The data for 10 species, were used to estimate Relation B, confirming that as Qx approached 1, kappa did approach 0 bar as predicted, and that as Qx approached 0, kappa approached approximately 750 bar, consistent with the a priori prediction of 500-1000 bar. The relations were also successfully tested using independent published data. An estimate of Qx is shown to be of considerable practical value in (a) converting Pb to water status and vice versa; (b) characterizing leaf morphology and composition; and (c) rationalizing quantitatively the functional classes of xerophytes, mesophytes and hygrophytes. The assumption of constant density inside the outer boundary of the non-gaseous material cannot be guaranteed, and when this is violated, our (or any other) interpretation of Pb is unreliable. Investigation of the conditions under which this assumption is invalid should be a high priority.     

Cardiac cells implanted into a cylindrical,vascularized chamber in vivo: pressure generation and morphology

We have previously described a model to implant dissociated cells into a cylindrical, vascularized bed in vivo to promote the formation of functional cardiac muscle constructs. We now investigate the cellular organization and the ability of the constructs to generate intra-luminal pressure. Primary cardiac cells were isolated from hearts of 2–3 day old rats, suspended in fibrin gel and inserted into the lumen of silicone tubing. The silicone tubing was then implanted around the femoral vessels in the groin region of recipient animals. After 3 weeks, the constructs were harvested, placed in an in vitro bath and cannulated via the incorporated femoral artery with a pressure transducer for evaluation of intra-luminal pressure dynamics. Histological evaluation showed the presence of a concentric ring of cardiac cells surrounding the femoral vessels. There was also a significant amount of collagen present around cardiac cells. In addition, we observed a significant amount of neovascularization of the explanted constructs. Electron microscopy showed the presence of longitudinally aligned fibers with a large number of gap junctions. Upon electrical stimulation of a single pulse (7 V, 1.2 ms), the constructs generated an intra-luminal pressure of 1.19 ± 0.45 mmHg (n = 6). In addition, we were able to electrically pace the constructs at frequencies of 0.5–5 Hz. A Starling behavior of the inverse relation between baseline pressure and twitch pressure was observed. Cardiac cells implanted for 3 weeks into the cylindrical vascularized bed formed a tissue construct that demonstrated many of the contractile properties and morphology expected of functioning cardiac tissues.     

Review. When to leave the brood chamber? Routes of dispersal in mites associated with burying beetles

Most nests of brood-caring insects are colonized by a rich community of mite species. Since these nests are ephemeral and scattered in space, phoresy is the principal mode of dispersal in mites specializing on insect nests. Often the mites will arrive on the nest-founding insect, reproduce in the nest and their offspring will disperse on the insect's offspring. A literature review shows that mites reproducing in the underground brood chambers of burying beetles use alternative routes for dispersal. For example, the phoretic instars of Poecilochirus spp. (Mesostigmata: Parasitidae) disperse early by attaching to the parent beetles. Outside the brood chamber, the mites switch host at carcasses and pheromone-emitting male beetles, where juvenile and mature burying beetles of several species congregate. Because they preferably switch to beetles that are reproductively active and use all species of burying beetles within their ranges, they have a good chance of arriving in a new brood chamber. Other mite associates of burying beetles (Alliphis necrophilus and Uropodina) disperse from the brood chamber on the beetle offspring. We suggest that these mites forgo the possible time gain of dispersing early on the parent beetles because their mode of attachment precludes host switching. Their phoretic instars, once attached, have to stay on their host and so only dispersing on the beetle offspring guarantees that they are present on reproducing burying beetles of the next season. The mites associated with burying beetles providean example of multiple solutions to one life history problem – how to find a new brood chamber for reproduction. Mites that have mobile phoretic instars disperse on the parent beetles and try to arrive in the next brood chamber by host switching. They are independent of the generation cycle of a single host and several generations of mites per host generation are possible. Mites that are constrained by their mode of attachment disperse on the beetle offspring and wait until their host becomes mature and reproduces. By doing this they synchronize their generation time with the generation time of their host species. Exp Appl Acarol 22: 621–631 © 1998 Kluwer Academic Publishers     

Content and structure of crew’s communicative behavior and gonadal hormone excretion during long-term chamber isolation of all-male international crew

The characteristics of psychophysiological adaptation to the simulation of the extended autonomous manned mission to Mars with limited resources and external communication were studied. The behavioral (communication with the external world) and physiological correlates of 105-day life and work in a pressurized compartment of the crew of six male subjects representing Russian and European space agencies under the conditions of sensory deprivation, confinement, monotony, and high autonomy were studied. Psychological, physiological, and biochemical (urinary cortisol and gonadal hormones) testing together with content analysis of the crew??s communication with the external world were made in parallel. The results confirmed, to a considerable extent, the preliminary hypothesis about the serious impact of stressful factors, including conflict, the significant decrease of the variety and scope of communication with the outer world, and social pressure on the verbal behavior and hormone levels in the future Martian crew. The correlation between the metabolism of the gonadal hormones and the volume, content, and creativity of the verbal human behavior during various stages of adaptation to the extended isolation in pressurized compartments has been discovered.     

High performance of CO2–temperature gradient chamber newly built for studying the global warming effect on a plant population

To study the effect of global warming concomitant with rising CO₂ on plant growth under field-like conditions, a CO₂–temperature gradient chamber (CTGC) was created from a standard temperature gradient chamber (TGC). Despite great daily and seasonal changes in ambient air temperature, the gradient of air temperatures was simulated by 5°C warmed conditions at the air outlet, rising at a rate of 1°C at 5 m intervals. Also, CO₂ concentrations were linearly increased from the air inlet to the outlet; those at the air inlet and 25 m distance from the air inlet were 372 p.p.m and 756 p.p.m. (doubled), respectively. The CTGC proved successful in biological experiments conducted through a full growth season in 1998. This apparatus will be useful for understanding plant response to simultaneous changes in CO₂ and temperature. Being a combination of a TGC and a CO₂ gradient chamber, the improved CTGC allows parameters and validation data sets to both be obtained for models contributing to plant growth, species composition, and global carbon flux analysis.     

Responses to ozone on Populus “Oxford” clone in an open top chamber experiment assessed before sunrise and in full sunlight

The effects of ambient levels of ozone and summer drought were assessed on a poplar clone (Populus maximowiczii Henry X P. × berolinensis Dippel — Oxford clone) in an open top chamber experiment carried out at the Curno facilities (Northern Italy). Chlorophyll (Chl) a fluorescence parameters (from both modulated and direct fluorescence) were assessed at different hours of the day (predawn, morning, midday, afternoon, and evening), from June to August 2008. This paper compares the results from predawn (PD, before sunrise) and afternoon (AN, in full sunlight) measurements, in order to evaluate the role of high sunlight as a factor influencing responses to ozone stress. Sunlight affected the maximum quantum yield of primary photochemistry (decrease of F_v/F_m) thus indicating photoinhibition. The effective quantum yield (Φ_PSII) and the photochemical quenching (q_P) were enhanced in the afternoon with respect to the predawn, whereas the nonphotochemical quenching (NPQ) was reduced. The effect of ozone was detected with fluorescence on well watered plants in the first week of July, before the onset of visible symptoms. As far as F_v/F_m are concerned, the differences between ozone-treated and control plants were statistically significant in the predawn, but not in the afternoon. Ozone exerted only minor effects on drought exposed plants because of the reduced stomatal ozone uptake, but effects on the IP phase of the fluorescence transient were observed also in drought-stressed plants.     

Morphometric analysis of the larval branchial chamber in the dragonfly Aeshna cyanea Müller (Insecta, Odonata, Anisoptera)

The aquatic larvae of anisopteran dragonflies possess tracheal gills located in the rectum. Using stereological methods, we estimated the morphometric diffusing capacity for oxygen (D(MO2)) across the gill epithelium, i.e., from rectal water to the gill tracheoles, in the larvae of Aeshna cyanea. A 271-mg larva has a total branchial surface area of approximately 12 cm(2). Tracheoles make up 6% of the epithelial volume of the gills; the harmonic mean thickness of the water-tracheolar diffusion barrier is 0.27 microm and consists mainly of cuticle. The calculated D(MO2) is 23.0 microl min(-1) g(-1) kPa(-1), which, using published values for oxygen consumption in a similar species, would result in a mean driving pressure of 0.2 kPa at rest and 1.3 kPa during activity. Since these driving pressures are similar to those reported for other arthropods, we conclude that the D(MO2) of the gill is not rate-limiting for aerobic metabolism in Aeshna cyanea larvae. J Morphol. 261:81-91, 2004.     

Detection of linked QTL for soybean brown stem rot resistance in ‘BSR 101’ as expressed in a growth chamber environment*

The objective of this study was to map the gene(s) conferring resistance to brown stem rot in the soybean cultivar BSR 101. A population of 320 recombinant inbred lines (RIL) was derived from a cross of BSR 101 and PI 437.654. Seedlings of each RIL and parent were inoculated by injecting stems with a suspension of spores and mycelia of Phialophora gregata, incubated in a growth chamber at 17°C, and assessed for resistance by monitoring the development of foliar and stem symptoms. The population also was evaluated with 146 RFLPs, 760 AFLPs, and 4 probes for resistance gene analogs (RGAs). Regression analysis identified a significant association between resistance and several markers on Linkage Group J of the USDA-ARS molecular marker linkage map. Interval analysis with Mapmaker QTL identified a major peak between marker RGA2V-1 and AFLP marker AAGATG152M on Linkage Group J. A second peak, associated only with stem symptoms, was identified between the RFLP B122I-1 and RGA2V-1, also on Linkage Group J. When composite interval mapping with QTL Cartographer was used, two linked QTL were identified with both foliar and stem disease assessment methods: a major QTL between AFLP markers AAGATG152E and ACAAGT260, and a minor QTL between RGA3I-3 and RGA3I-2. These results demonstrate that composite interval mapping gives increased precision over interval mapping and is capable of distinguishing two linked QTL. The minor QTL associated with the cluster of RGA3I loci is of special interest because it is the first example of a disease resistance QTL associated with a resistance gene analog.     

Human keratinocytes in culture exhibit no response when exposed to short duration, low amplitude, high frequency (900 MHz) electromagnetic fields in a reverberation chamber

We exposed normal human epidermal keratinocytes to short duration, high frequency, and low amplitude electromagnetic fields, similar to that used by mobile phone technologies. We paid particular attention to the control of the characteristics of the electromagnetic environment generated within a mode stirred reverberation chamber (statistical homogeneity and isotropy of the field and SAR distribution). Two non‐thermal exposure conditions were tested on the epidermal cells: 10‐min exposure with a field amplitude of 8 V/m, and 30 min with 41 V/m. Corresponding specific absorption rates ranged from 2.6 to 73 mW/kg (continuous wave, 900 MHz carrier frequency). We collected RNA from cells subjected to these conditions and used it for a large‐scale microarray screening of over 47000 human genes. Under these conditions, exposure of keratinocytes to the electromagnetic field had little effect; only 20 genes displayed significant modulation. The expression ratios were very small (close to 1.5‐fold change), and none of them were shared by the two tested conditions. Furthermore, those assayed using polymerase chain reaction did not display significant expression modulation (overall mean of the exposed samples: 1.20 ± 0.18). In conclusion, the data presented here show that cultured keratinocytes are not significantly affected by EMF exposure. Bioelectromagnetics 32:302–311, 2011. © 2010 Wiley‐Liss, Inc.     

Development of the “plankton emergence trap/chamber (PET Chamber)”, a new sampling device to collect in situ germinating cells from cysts of microalgae in surface sediments of coastal waters

A simple device was developed to collect germinating cells from settled cysts of microalgae on the sea bottom. This new device – the plankton emergence trap chamber (PET Chamber) – made of a clear acrylic plastic, consists of a top cylinder and a base plate attached to a bottom cylinder. After plugging the bottom cylinder with sediment collected from in situ sea bottom and filling the top cylinder with filtered seawater, the PET Chamber attached to a platform specially made for submerging the chambers is placed on the seafloor. An adequate area of the lateral side of the top cylinder is opened and covered by a plankton net (10 μm mesh) to allow a water exchange between the inside and outside the cylinder. Thus, the PET Chamber can replicate in situ environments, such as temperature, irradiance and dissolved oxygen. Using the PET Chamber, we have succeeded in collecting germinating cells and estimating the germination flux (cells m⁻² day⁻¹) of two dinoflagellates, Alexandrium catenella and Scrippsiella spp., in Ago Bay, Japan. Mircoscopic observations on the samples collected from July to October 2003 revealed fluxes of 124–2022 and 622–3732 cells m⁻² day⁻¹ in A. catenella and Scrippsiella spp., respectively. The data indicate that the new device can detect variations in the number of germinating cells of dinoflagellates. Its sampling ability, coupled with simplicity in deployment and retrieval procedures, can allow one to monitor the in situ emergence from/on the surface sediments of various organisms, that possess resting stages during their life histories, in various coastal waters.