In situ measurement of native extracellular matrix strain.

Cells directly interact with the extracellular matrix (ECM) in their microenvironment; however, the mechanical properties of the networks at this scale are not well defined. This work describes a method to quantify ECM network strain in situ after the application of a known load. Visualization of the ECM in the native 3D organization is facilitated using murine embryos and a novel decellularization method. During embryonic development, the ECM architecture is less dense making it easier to visualize and manipulate. Briefly, embryonic day (E)14.5 forelimbs were harvested and incubated in an acrylamide-based hydrogel mixture to maintain the 3D architecture of the ECM during decellularization. After decellularization, forelimbs were stained for fibrillin-2 and proteoglycans to visualize different networks. Samples were imaged, before and after the application of a static load, using confocal microscopy. A MATLAB-based fast iterative digital volume correlation algorithm was used to quantify network displacement fields by comparing the reference and compressed z-stacks. We observed that the amount of Green-Lagrange strain experienced by different proteins was dependent on whether the sub-region analyzed was located within cartilage or the adjacent connective tissue. The combination of these experimental and computational methods will enable the development of constitutive equations that describe the material behavior of ECM networks. In the future, this information has the potential to improve the fabrication of physiologically relevant scaffolds by establishing mechanical guidelines for microenvironments that support beneficial cell-ECM interactions.

Fast sorption measurements of volatile organic compounds on building materials: Part 1 - Methodology developed for field applications.

A Proton Transfer Reaction-Mass Spectrometer (PTR-MS) has been coupled to the outlet of a Field and Laboratory Emission Cell (FLEC), to measure volatile organic compounds (VOC) concentration during a sorption experiments (Rizk et al., this issue) [1]. The limits of detection of the PTR-MS for three VOCs are presented for different time resolution (2, 10 and 20 s). The mass transfer coefficient was calculated in the FLEC cavity for the different flow rates. The concentration profile obtained from a sorption experiment performed on a gypsum board and a vinyl flooring are also presented in comparison with the profile obtained for a Pyrex glass used as a material that do not present any sorption behavior (no sink). Finally, the correlation between the concentration of VOCs adsorbed on the surface of the gypsum board at equilibrium (Cse ) and the concentration of VOCs Ce measured in the gas phase at equilibrium is presented for benzene, C8 aromatics and toluene.

Hemoconcentration induced by exercise: Revisiting the Dill and Costill equation.

The Dill and Costill equation is used to estimate the exercise-induced hemoconcentration. However, this calculation requires drawing an extra whole-blood sample, which cannot be frozen and has to be analyzed with dedicate instrumentation in a relative short time. The aim of the present study was to explore the usefulness of some serum biochemical parameters to estimate hemoconcentration induced by exhaustive exercise. Fourteen healthy male subjects (19-34 years) performed a15-min running test at 110% of anaerobic threshold speed. Hemoglobin, hematocrit, brain natriuretic peptide (BNP), creatinine, gamma-glutamyltransferase (GGT), total-proteins, albumin, total calcium (Ca), K(+), Na(+), and Cl(-) were determined in blood samples taken before, after exercise, and after a 30-min recovery period. Plasma volume loss (ΔPV) was calculated by Dill and Costill equation. At post-exercise and after recovery, the percentage increments of total-proteins, albumin, GGT and Ca correlated significantly with ΔPV. Bland-Altman analyses showed that correcting BNP, creatinine, and K(+) concentration by Ca percentage increments yield biases and limits of agreement that are acceptable when compared with Dill and Costill equation correction. Ca concentration may be used as a hemoconcentration biomarker in high-intensity exercise, which would allow scientists and physicians avoid extra costs, facilitate in-field research, and delayed estimation of hemoconcentration using stored serum samples.

Optical clearing in collagen- and proteoglycan-rich osteochondral tissues.

OBJECTIVE: Recent developments in optical clearing and microscopy technology have enabled the imaging of intact tissues at the millimeter scale to characterize cells via fluorescence labeling. While these techniques have facilitated the three-dimensional (3D) cellular characterization within brain and heart, study of dense connective tissues of the musculoskeletal system have been largely unexplored. Here, we quantify how optical clearing impacted the cell and tissue morphology of collagen-, proteoglycan-, and mineral-rich cartilage and bone from the articulating knee joint. METHODS: Water-based fructose solutions were used for optical clearing of bovine osteochondral tissues, followed by imaging with transmission and confocal microscopy. To confirm preservation of tissue structure during the clearing process, samples were mechanically tested in unconfined compression and visualized by cryo-SEM. RESULTS: Optical clearing enhanced light transmission through cartilage, but not subchondral bone regions. Fluorescent staining and immunolabeling was preserved through sample preparations, enabling imaging to cartilage depths five times deeper than previously reported, limited only by the working distance of the microscope objective. Chondrocyte volume remained unchanged in response to, and upon the reversal, of clearing. Equilibrium modulus increased in cleared samples, and was attributed to exchange of interstitial fluid with the more viscous fructose solution, but returned to control levels upon unclearing. In addition, cryo-SEM-based analysis of cartilage showed no ultrastructural changes. CONCLUSION: We anticipate large-scale microscopy of diverse connective tissues will enable the study of intact, 3D interfaces (e.g., osteochondral) and cellular connectivity as a function of development, disease, and regeneration, which have been previously hindered by specimen opacity.

Experimental performances study of a transportable GC-PID and two thermo-desorption based methods coupled to FID and MS detection to assess BTEX exposure at sub-ppb level in air.

BTEX compounds are of particular interest, above all benzene because it is a carcinogenic compound for which guideline value in European indoor environments is set to be 1.6 ppb. Therefore, the detection of such relatively low value requires the use of particularly sensitive analytical techniques. Several existing chromatographic techniques, such as fast and transportable Gas Chromatograph with Photoionization Detection (GC-PID) or sedentary chromatographic-based techniques equipped with a thermo-desorption device (ATD) and coupled to either Flame Ionization Detection (FID) or Mass Spectrometry (MS), can quantify benzene and its derivatives at such low levels. These instruments involve different injection modes, i.e. on-line gaseous sampling or thermo-desorption of adsorbent tubes spiked with liquid or gas samples. In this study, the performances of 3 various analytical techniques mentioned above were compared in terms of sensitivity, linearity, accuracy and repeatability for the 6 BTEX. They were also discussed related to their analyses time consumption or transportability. The considered analytical techniques are ATD-GC-FID, ATD-GC-MS where both full scan and SIM modes were tested and a transportable GC-PID. For benzene with on-line injection, Limits of Detection (LOD) were significantly below the European guideline with values of 0.085, 0.022, 0.007 and 0.058 ppb for ATD-GC-FID, ATD-GC-MS in a full scan mode, ATD-GC-MS in an SIM mode and transportable GC-PID, respectively. LOD obtained with adsorbent tubes spiked with liquid standards were approximately in the same order of magnitude.

Specific accumulation of CYP94A1 transcripts after exposure to gaseous benzaldehyde: induction of lauric acid ω-hydroxylase activity in Vicia sativa exposed to atmospheric pollutants.

The effects of air pollutants such as aldehydes, ozone, nitrogen dioxide and benzene on fatty acid ω-hydroxylase activity in Vicia sativa microsomes have been investigated. Four days old etiolated V. sativa seedlings were exposed to different concentrations of selected pollutants for varying exposure times. Growing etiolated V. sativa seedlings in air containing the gaseous benzaldehyde (150 nM) led to an 8-fold enhancement of lauric acid ω-hydroxylase activity in microsomes of treated plants compared to controls grown in pure air (96 ± 10 versus 12 ± 2 pmol/min/mg protein, respectively). The induction increased with increasing gas phase concentrations (10-1300 nM) and the maximum of activity was measured after 48 h of exposure. Northern blot analysis revealed that this induction occurred via transcriptional activation of the gene coding for CYP94A1. The absence of CYP94A2 and CYP94A3 transcription activation together with the missing effect on epoxide hydrolases activities indicate the specificity of CYP94A1 induction by benzaldehyde. Exposure to nitrogen dioxide, ozone and formaldehyde also stimulated lauric acid ω-hydroxylases activity while exposure to benzene did not show any effect.

Uptake measurements of acetaldehyde on solid ice surfaces and on solid/liquid supercooled mixtures doped with HNO3 in the temperature range 203-253 K.

Uptake of acetaldehyde on ice surfaces has been investigated over the temperature range 203-253 K using a coated wall flow tube coupled to a mass spectrometric detection. The experiments were conducted on pure ice surfaces and on liquid/solid ice mixture both doped with nitric acid (0.063, 0.63, and 6.3 wt %). Uptake of acetaldehyde on these surfaces was always found to be totally reversible whatever the experimental conditions were. The number of acetaldehyde molecules adsorbed per surface unit was conventionally plotted as a function of acetaldehyde concentration in the gas phase. Although the amounts of acetaldehyde adsorbed on solid ice surfaces (pure and HNO(3)-doped ice) were approximately similar and rather limited, the number of acetaldehyde molecules taken up on the HNO(3)-doped solid ice/liquid mixtures are significantly higher, up to 1 or 2 orders of magnitudes compared to pure ice surfaces. At 213 K for example and for low concentrations of acetaldehyde (<1 x 10(13) molecule cm(-3)), the amount of acetaldehyde molecules taken up on solid/liquid doped surfaces is 3.3 and 8.8 times higher than those measured on pure ice respectively for 0.063 and 0.63 wt % of HNO(3). The huge quantities of acetaldehyde taken up by liquid-/solid-doped mixtures are likely dissolved in the nonhomogeneous liquid part of the surfaces according to the Henry's law equilibrium. As a consequence, up to about 10% of acetaldehyde may be scavenged by supercooled liquid droplets of convective clouds in the upper troposphere.

Uptake measurements of ethanol on ice surfaces and on supercooled aqueous solutions doped with nitric acid between 213 and 243 K.

Uptake of ethanol either on pure frozen ice surfaces or supercooled solutions doped with HNO3 (0.63 and 2.49 wt %) has been investigated using a coated wall flow tube coupled to a mass spectrometric detection. The experiments were conducted over the temperature range of 213-243 K. Uptake of ethanol on these surfaces was always found to be totally reversible whatever were the experimental conditions. The number of ethanol molecules adsorbed per surface unit was conventionally plotted as a function of ethanol concentration in the gas phase and subsequently analyzed using Langmuir's model. The amount of ethanol molecules taken up on nitric acid doped-ice surfaces was found to increase largely with increasing nitric acid concentrations. For example at 223 K, and for an ethanol gas-phase concentration of 1x10(13) molecules cm3, the number of adsorbed molecules are (in units of molecules cm-2): approximately 1.3x10(14) on pure ice; approximately 1.4x10(15) on ice doped with HNO3 0.63 wt %; approximately 7.5x10(15) on ice doped with HNO3, 2.49 wt %, i.e. 60 times larger than on pure ice. Since, according to the shape of the isotherms, the adsorption did not proceed beyond monolayer coverage, the enormous increase of ethanol uptake was explained by considering its dissolution in either a supercooled liquid layer (T<230 K) or a liquid solution (T>230 K). The formation of both was indeed favored by the presence of the HNO3. Our experimental results suggest that the amount of ethanol dissolved in such supercooled solutions follows Henry's law and that the Henry's law constants at low temperatures, i.e., 223-243 K, can be estimated by extrapolation from higher temperatures. Such supercooled solutions which exist in the troposphere either in deep convective clouds or in mixed clouds for temperature above 233 K, might be responsible for the scavenging of large amounts of soluble species, such as nitric and sulfuric acids, oxygenated VOCs including alcohols, carboxylic acids, and formaldehyde.

Inhaled formaldehyde exposure: effect on bronchial response to mite allergen in sensitized asthma patients.

BACKGROUND: Formaldehyde, an indoor air pollutant, is known to be an irritant and an etiologic factor in occupational asthma. An epidemiologic study suggests that it may also increase the risk of childhood asthma for concentrations above 60 microg/m(3). AIM: To evaluate the influence of pre-exposure to low-dose formaldehyde (100 microg/m(3) in 30 min according to the World Health Organization's recommended maximum value for indoor environments) on bronchial response to Dermatophagoides pteronyssinus. METHOD: Nineteen asthmatic subjects were included. Each subject underwent a mite allergen bronchial challenge test immediately after a standardized exposure in a chamber to formaldehyde or air (random order). Induced sputum were collected 24 h before and after mite challenge. RESULTS: After formaldehyde inhalation, patients developed an immediate bronchial response at a significantly lower dose of mite allergen than after air exposure (the geometric mean PD(20) for Der p 1 was 34.3 ng after formaldehyde and 45.4 ng after placebo, P = 0.05). The late-phase reaction, expressed as the maximum fall in forced expiratory volume in 1 s (FEV(1)) from baseline, was significantly higher after formaldehyde (15%vs 11%, P = 0.046). CONCLUSION: Our study demonstrated that exposure to low levels of formaldehyde significantly enhanced bronchial responsiveness to mite allergen in mite-sensitized subjects with asthma.

[The bronchial response to inhaled formaldehyde]. / Le formaldéhyde inhalé et la réponse bronchique.

INTRODUCTION: Formaldehyde is an ubiquitous indoor chemical polutant. Occupational exposure to high concentrations has revealed its irritant and allergenic potential. Nevertheless, domestic exposure to low concentrations may also have an effect on respiratory health in a non-specific way, just as has been found for other pollutants. STATE OF KNOWLEDGE: Potentiation of the response to allergens has been observed in animals and children. This effect has also been found on respiratory symptoms, with a 39% increase in the risk of asthma for a domiciliary exposure of more than 60 microgrammes.m(-3). We have recently been able to show, in a study with asthmatics sensitised to house dust mite, that the response to allergen provocation was increased following a 30 minutes exposure to 100 microgrammes.m(-3) formaldehyde. VIEWPOINT AND CONCLUSIONS: All the data show that mild exposure to formaldehyde in the home is sufficient to provoke sensitisation and also an aggrevation of symptoms in patients with allergic asthma. Taking into account the published evidence it is advisable that the concentrations of formaldehyde in domestic products should be made known in order to improve domiciliary air quality.

Atmospheric fate of dichlorvos: photolysis and OH-initiated oxidation studies.

The OH-initiated oxidation of dichlorvos (a widely used insecticide) has been investigated under atmospheric conditions at the large outdoor European photoreactor (EUPHORE) in Valencia, Spain. The rate constant of OH reaction with dichlorvos, k, was measured by using a conventional relative rate technique where 1,3,5-trimethylbenzene (TMB) and cyclohexane were taken as references. With the use of the rate constants of 5.67 x 10(-11) and of 6.97 x 10(-12) cm3 molecule(-1) s(-1) for the reactions OH + TMB and OH + cyclohexane, respectively, the resulting value of the OH reaction rate constant with dichlorvos was derived to be k = (2.6 +/- 0.3) x 10(-11) cm3 molecule(-1) s(-1). The tropospheric lifetime of dichlorvos with respect to reaction with OH radical has been estimated to be around 11 h. The major carbon-containing products observed for the OH reaction with dichlorvos in air under sunlight condition were phosgene and carbon monoxide. The formation of a very stable toxic primary product such as phosgene associated with the relatively short lifetime of dichlorvos may make the use of this pesticide even more toxic for humans when released into the atmosphere.

Adsorption of acetic acid on ice: experiments and molecular dynamics simulations.

Adsorption study of acetic acid on ice surfaces was performed by combining experimental and theoretical approaches. The experiments were conducted between 193 and 223 K using a coated wall flow tube coupled to a mass spectrometric detection. Under our experimental conditions, acetic acid was mainly dimerized in the gas phase. The surface coverage increases with decreasing temperature and with increasing concentrations of acetic acid dimers. The obtained experimental surface coverages were fitted according to the BET theory in order to determine the enthalpy of adsorption deltaH(ads) and the mololayer capacity N(M(dimers)) of the acetic acid dimers on ice: deltaH(ads) = (-33.5 +/- 4.2) kJ mol(-1), N(M(dimers)) = (l1.27 +/- 0.25) x 10(14) dimers cm(-2). The adsorption characteristics of acetic acid on an ideal ice I(n)(0001) surface were also studied by means of classical molecular dynamics simulations in the same temperature range. The monolayer capacity, the configurations of the molecules in their adsorption sites, and the corresponding adsorption energies have been determined for both acetic acid monomers and dimers, and compared to the corresponding data obtained from the experiments. In addition, the theoretical results show that the interaction with the ice surface could be strong enough to break the acetic acid dimers that exist in the gas phase and leads to the stabilization of acetic acid monomers on ice.

Adsorption study of acetone on acid-doped ice surfaces between 203 and 233 K.

Adsorption studies of acetone on pure ice surfaces obtained by water freezing or deposition or on frozen ice surfaces doped either with HNO3 or H2SO4 have been performed using a coated wall flow tube coupled to a mass spectrometric detection. The experiments were conducted over the temperature range 203-233 K and freezing solutions containing either H2SO4 (0.2 N) or HNO3 (0.2-3 N). Adsorption of acetone on these ice surfaces was always found to be totally reversible whatever were the experimental conditions. The number of acetone molecules adsorbed per ice surface unit N was conventionally plotted as a function of acetone concentration in the gas phase. For the same conditions, the amount of acetone molecules adsorbed on pure ice obtained by deposition are about 3-4 times higher than those measured on frozen ice films, H2SO4-doped ice surfaces lead to results comparable to those obtained on pure ice. On the contrary, N increases largely with increasing concentrations of nitric acid in ice surfaces, up to about 300 times under our experimental conditions and for temperatures ranging between 213 and 233 K. Finally, the results are discussed and used to reestimate the partitioning of acetone between the ice and gas phases in clouds of the upper troposphere.

Kinetic Studies of OH Reactions with a Series of Methyl Esters.

Absolute rate constants have been measured for the gas-phase reactions of hydroxyl radicals with a series of methyl esters: methyl propionate (k 1), methyl butyrate (k 2), methyl valerate (k 3), and methyl caproate (k 4). Experiments were carried out using the pulsed laser photolysis-laser induced fluorescence technique over the temperature range 253-372 K. The obtained kinetic data were used to derive the following Arrhenius expressions: k 1 = (1.45 ± 0.42) × 10-12 exp[-(148 ± 86)/T]; k 2 = (0.96 ± 0.29) × 10-12 exp[(380 ± 91)/T]; k 3 = (1.37 ± 0.64) × 10-12 exp[(401 ± 142)/T]; k 4 = (2.46 ± 1.04) × 10-12 exp[(326 ± 130)/T] (in units of cm3 molecule-1 s-1). At room temperature, the rate constants obtained (in units of 10-12 cm3 molecule-1 s-1) were as follows: methyl propionate (0.83 ± 0.09); methyl butyrate (3.30 ± 0.25); methyl valerate (4.83 ± 0.55); methyl caproate (7.15 ± 0.70). Our results are compared with the previous determinations and discussed in terms of structure-activity relationships.