Ideonella sakaiensis Can Metabolize Bisphenol A as a Carbon Source.

Bisphenol A and its analogues represent a significant environmental and public health hazard, particularly affecting the endocrine systems of children and newborns. Due to the growing need for non-pathogenic biodegradation microbial agents as environmentally friendly and cost-effective solutions to eliminate endocrine disruptors, this study aimed to investigate the degradation of bisphenol A by Ideonella sakaiensis, based on its currently understood unique enzymatic machinery that is already well known for degrading polyethylene terephthalate. The present study provides novel insights into the metabolic competence and growth particularities of I. sakaiensis. The growth of I. sakaiensis exposed to bisphenol A exceeded that in the control conditions, starting with 72 h in a 70% nutrient-rich medium and starting with 48 h in a 100% nutrient-rich medium. Computational modeling showed that bisphenol A, as well as its analogue bisphenol S, are possible substrates of PETase and MHETase. The use of bisphenol A as a carbon and energy source through a pure I. sakaiensis culture expands the known substrate spectra and the species' potential as a new candidate for bisphenol A bioremediation processes.

Rapid Environmental Impact Assessment of Penicillin G in a Veterinary Product Using an Original Software Method and Monitoring by SPE-Online-UHPLC-MS/MS.

Veterinary antibiotics have become a major concern due to potential environmental effects. This study presents an investigation of the exposure and environmental effects of the veterinary medicinal product in powder form, administered in the drinking water of piglets, chickens and turkeys, containing 250 mg/g penicillin G (benzylpenicillin potassium), performed according to the European Medicines Agency (EMEA) guideline and the results obtained by an analytical method based on online solid-phase extraction, ultra-high performance liquid chromatography coupled to a triple quadrupole mass spectrometer (SPE-online-UHPLC-MS/MS). The study presents the determination of the environmental risk and through an original, interactive, fast software method, created on the basis of a proprietary calculation algorithm that goes through all the prescriptions and recommendations of the EMEA guide. The results demonstrated that the concentration value for penicillin G determined in surface water by SPE-online-UHPLC-MS/MS is much lower than that predicted by calculation (predictable concentration in surface water, PECsurface water = 37.66 µg/L and the concentration SPE-online-UHPLC-MS/MS = 0.032 µg/L). Both results lead to a sub-unit risk quotient (R) indicating that the treatment carried out with the considered veterinary product does not present any risk to the environment.

Variability of PM10 level with gaseous pollutants and meteorological parameters during episodic haze event in Malaysia: Domestic or solely transboundary factor?

Haze has become a seasonal phenomenon affecting Southeast Asia, including Malaysia, and has occurred almost every year within the last few decades. Air pollutants, specifically particulate matter, have drawn a lot of attention due to their adverse impact on human health. In this study, the spatial and temporal variability of the PM10 concentration at Kelang, Melaka, Pasir Gudang, and Petaling Jaya during historic haze events were analysed. An hourly dataset consisting of PM10, gaseous pollutants and weather parameters were obtained from Department of Environment Malaysia. The mean PM10 concentrations exceeded the stipulated Recommended Malaysia Ambient Air Quality Guideline for the yearly average of 150 µg/m3 except for Pasir Gudang in 1997 and 2005, and Petaling Jaya in 2013. The PM10 concentrations exhibit greater variability in the southwest monsoon and inter-monsoon periods at the studied year. The air masses are found to be originating from the region of Sumatra during the haze episodes. Strong to moderate correlation of PM10 concentrations was found between CO during the years that recorded episodic haze, meanwhile, the relationship of PM10 level with SO2 was found to be significant in 2013 with significant negatively correlated relative humidity. Weak correlation of PM10-NOx was measured in all study areas probably due to less contribution of domestic anthropogenic sources towards haze events in Malaysia.

Flexible Investment Casting Wax Patterns for 3D-Printing: Their Rheological and Mechanical Characterizations.

The mechanical and rheological characterizations of flexible investment casting patterns capable of 3D printing are reported. The wax pattern was composed of microcrystalline hydrocarbon wax (DMW7478), Piccotex 75 (a copolymer of α-methyl-styrene and vinyl toluene with a 75/25 molar ratio, respectively) and Escorene (a copolymer of ethylene and vinyl acetate with a 72/28 mass ratio, respectively). It was found that in order to obtain a wax pattern with appreciable mechanical properties, the content of the microcrystalline hydrocarbon wax in these blends should not exceed 30% (m/m). Thus, a series of patterns with 28% (m/m) wax and varying Piccotex and Escorene contents spanning from 0 to 72% (m/m) was prepared. The dependence of the dynamic viscosities of the wax patterns on the composition was described using a stretched exponential model, whereas their variations with the temperature were interpreted in terms of the Arrhenius-Guzman equation. Furthermore, the slopes of the lines fitted to the viscosity versus temperature curves at the pour point decreased linearly with the Piccotex content. Non-Newtonian changes in the shear stress with the shear rate and shear stress crystallization were observed at temperatures near the pour points. The mechanical properties were evaluated using the uniaxial tensile mode and by three-point bending experiments. It was found that the stress (σ) versus the relative elongation (ε) curves can effectively be rendered by means of the standard linear solid (SLS) viscoelastic model. In addition, it was also established that the Young's modulus varied according to a sigmoid-type curve with the piccotex content, while the yield stress decreased linearly with the concentration of Piccotex. In addition, based on the spooling suitability and printability, the patterns were rated and it was found that the most appropriate wax pattern compositions for 3D printing were those which contained 30% (m/m) and 35% (m/m) Piccotex.

Overall assessment of surface water quality in the Lower Danube River.

Water is an important receptor of environmental problems generated by anthropogenic activities, the water quality being affected by their magnitude and impact as well as by climate change. Given the international and cross-border nature of the Danube River and being aware that the pollution risk assessment is one of the key elements of the ecology and health of the people in its region, this paper assessed the state of water quality in the Lower Danube River in accordance with Water Framework Directive (Directive 2000/60/EC). The intensive monitoring program applied by the National Institute for Research and Development in Environmental Protection (INCDPM) took place during the period 2011-2017, in which approximately 1500 samples were taken and processed from 10 monitoring sites and the quality indicators stipulated in M.O. 161/2006 were analysed according to the standards in force. In this monitoring area (km 375-km 175), hydrotechnical works have been done in order to improve navigation conditions. Therefore, on the basis of the data obtained from the monitoring campaigns, multivariate statistical methods of water quality assessment were applied on the complex set of data, and at the same time, the multiparametric quality index (ICPM) was applied, an index of global comparative assessment of water quality over historical trends, developed by INCDPM. Following the evaluation of the results, the water quality in the Lower Danube River was characterized as moderately polluted and corresponds to Class III of surface water quality. The ecosystem approach indicates that the values of the monitored indicators did not correspond to the target values according to the Water Framework Directive.

Cell physiological effects of glass ionomer cements on fibroblast cells.

The cytotoxicity of glass ionomer cements (GICs) was investigated using a novel, cost-effective, easy-to-perform and standardized test. GIC rings were made using in-house designed, custom-made moulds under sterile conditions; 10 with Fuji Equia and 10 with Fuji Triage capsules, placed in direct contact with primary human gingival fibroblasts (HGF) and immortalized human fibroblasts (HFF1). On day 1, 4, 14 and 21, an AlamarBlue® (resazurin) assay was completed towards determining the effects of the GICs on metabolic activities of the cells, whilst cell morphology was examined by light microscopy. The influence of the compounds released from the GIC rings on cell physiological effects (viability, proliferation and adhesion) during 24 h incubation was further investigated by impedimetry. Result trends obtained from this battery of techniques were complementary. At 100 v/v% concentration, the released compounds from Equia were strongly cytotoxic, while at lower concentration (0, 4, 20 v/v%) they were not cytotoxic. In contrast, Triage elicited only slightly transient cytotoxicity. The method proposed has been proved as being efficient, reliable and reproducible and may be useful in quick testing of the cytotoxicity of similar biomaterials by using an immortalized cell line.

Designed Polyurethanes for Potential Biomedical and Pharmaceutical Applications: Novel Synthetic Strategy for Preparing Sucrose Containing Biocompatible and Biodegradable Polyurethane Networks.

In this paper the preparation and detailed characterization of designed polyurethanes (SPURs) are reported for potential biological, biomedical and/or pharmaceutical applications. Importantly, in order to fulfill these goals all reactants and solvents used were selected according to the proposal of EUR-8 Pharmacopoeia. For the synthesis, a novel strategy was introduced and elaborated. A series of SPUR samples was prepared from poly(ε-caprolactone)-diol, 1,6-hexamethylene diisocyanate and sucrose as a chain extender/crosslinking agent to obtain sucrose containing polyurethanes. In addition, the mol ratios of the sucrose were varied within an order of magnitude. The prepolymers and the products of the syntheses were investigated by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and infrared spectroscopy (IR), respectively. It was found that the reactivity of the eight free hydroxyl groups of sucrose are different, and after curing the SPUR samples at 60 °C no free isocyanate groups can be observed. Furthermore, swelling experiments performed with various solvents of different polarities revealed that the highest degree of swelling took place in dimethyl-sulfoxide. However, low degrees of swelling were recognized in water and hexane. It is important to note that the gel contents were around 90% in all cases, which demonstrate that the crosslinking was almost complete. In addition, the kinetics of swelling were also evaluated and successfully modeled. The crosslink densities were calculated from the data of the swelling experiments by means of the Flory-Rehner equation. Unexpectedly, it was found that the crosslink density decreased with the increasing sucrose content also in line with the results obtained by relaxation modulus experiments and dynamic mechanical analysis (DMA). The Tg and Tm of SPUR samples, determined from DSC and DMA measurements, were around -57 °C and 27 °C, respectively. According to the mechanical tests the SPUR samples showed high elongation at break values, i.e., high flexibilities. Furthermore, the stress-strain curves were also modeled and discussed.

Gas-Phase Interaction of Anions with Polyisobutylenes: Collision-Induced Dissociation Study and Quantum Chemical Modeling.

The gas-phase interaction of anions including fluoride, chloride, bromide, iodide, ethyl sulfate, chlorate, and nitrate with polyisobutylene (PIB) derivatives was studied using collision-induced dissociation (CID). The gas-phase adducts of anions with PIBs ([PIB + anion](-)) were generated from the electrosprayed solution of PIBs in the presence of the corresponding anions. The so-formed adducts subjected to CID showed a loss of anion at different characteristic collision energies, thus allowing the study of the strength of interaction between the anions and nonpolar PIBs having different end-groups. The values of characteristic collision energies (the energy needed to obtain 50% fragmentation) obtained by CID experiments correlated linearly with the binding enthalpies between the anion and PIB, as determined by density functional theory calculations. In the case of halide ions, the critical energies for dissociation, that is, the binding enthalpies for [PIB + anion](-) adducts, increased in the order of I(-) < Br(-) < Cl(-) < F(-). Furthermore, it was found that the binding enthalpies for the adducts formed with halide ions decreased approximately with the square radius of the halide ion, suggesting that the strength of interaction is mainly determined by the "surface" charge density of the halide ion. In addition, the characteristic collision energy versus the number of isobutylene units revealed a linear dependence.

Can Nonpolar Polyisobutylenes be Measured by Electrospray Ionization Mass Spectrometry? Anion-Attachment Proved to be an Appropriate Method.

Polyisobutylenes (PIBs) with different end-groups including chlorine, exo-olefin, hydroxyl, and methyl prepared from aliphatic and aromatic initiators were studied by electrospray ionization mass spectrometry (ESI-MS). Independently of the end-groups, presence or absence of aromatic initiator moiety, these PIB derivatives were capable of forming adduct ions with NO3 (-) and Cl(-) ions, thus allowing the direct characterization of these compounds in the negative ion mode of ESI-MS. To obtain [PIB + NO3](-) and [PIB + Cl](-) adduct ions with appreciable intensities, addition of polar solvents such as acetone, 2-propanol, or ethanol to the dichloromethane solution of PIBs was necessary. Furthermore, increasing both the polarity (by increasing the acetone content) and the ion-source temperature give rise to enhanced intensities for both [PIB + NO3](-) and [PIB + Cl](-) ions. Energy-dependent collision induced dissociation studies (CID) revealed that increasing the collision voltages resulted in the shift of the apparent molecular masses to higher ones. CID studies also showed that dissociation of the [PIB + Cl](-) ions requires higher collision energy than that of [PIB + NO3](-). In addition, Density Functional Theory calculations were performed to gain insights into the nature of the interactions between the highly non-polar PIB chains and anions NO3 (-) and Cl(-) as well as to determine the zero-point corrected electronic energies for the formation of [PIB + NO3](-) and [PIB + Cl](-) adduct ions.

Serum osteoprotegerin is associated with pulse pressure in kidney transplant recipients.

Pulse pressure (PP) reflects increased large artery stiffness, which is caused, in part, by arterial calcification in patients with chronic kidney disease. PP has been shown to predict both cardiovascular and cerebrovascular events in various patient populations, including kidney transplant (KTX) recipients. Osteoprotegerin (OPG) is a marker and regulator of arterial calcification, and it is related to cardiovascular survival in hemodialysis patients. Here we tested the hypothesis that OPG is associated with increased pulse pressure. We cross-sectionally analyzed the association between serum OPG and PP in a prevalent cohort of 969 KTX patients (mean age: 51 +/- --13 years, 57% male, 21% diabetics, mean eGFR 51 +/- 20 ml/min/1.73 m2). Independent associations were tested in a linear regression model adjusted for multiple covariables. PP was positively correlated with serum OPG (rho = 0.284, p < 0.001). Additionally, a positive correlation was seen between PP versus age (r = 0.358, p < 0.001), the Charlson Comorbidity Index (r = 0.232, p < 0.001), serum glucose (r = 0.172, p < 0.001), BMI (r = 0.133, p = 0.001) and serum cholesterol (r = 0.094, p = 0.003). PP was negatively correlated with serum Ca, albumin and eGFR. The association between PP and OPG remained significant after adjusting for multiple potentially relevant covariables (beta = 0.143, p < 0.001). We conclude that serum OPG is independently associated with pulse pressure in kidney transplant recipients.

Direct analysis in real time mass spectrometry (DART-MS) of highly non-polar low molecular weight polyisobutylenes.

Low molecular weight polyisobutylenes (PIB) with chlorine, olefin and succinic acid end-groups were studied using direct analysis in real time mass spectrometry (DART-MS). To facilitate the adduct ion formation under DART conditions, NH4 Cl as an auxiliary reagent was deposited onto the PIB surface. It was found that chlorinated adduct ions of olefin and chlorine telechelic PIBs, i.e. [M + Cl]- up to m/z 1100, and the deprotonated polyisobutylene succinic acid [MH]- were formed as observed in the negative ion mode. In the positive ion mode formation of [M + NH4 ]+ , adduct ions were detected. In the tandem mass (MS/MS) spectra of [M + Cl]- , product ions were absent, suggesting a simple dissociation of the precursor [M + Cl]- into a Cl- ion and a neutral M without fragmentation of the PIB backbones. However, structurally important product ions were produced from the corresponding [M + NH4 ]+ ions, allowing us to obtain valuable information on the arm-length distributions of the PIBs containing aromatic initiator moiety. In addition, a model was developed to interpret the oligomer distributions and the number average molecular weights observed in DART-MS for PIBs and other polymers of low molecular weight. Copyright © 2015 John Wiley & Sons, Ltd.

Collision-induced dissociation study of poly(2-ethyl-2-oxazoline) using survival yields and breakdown curves.

Poly(2-ethyl-2-oxazoline), a synthetic polymer was analysed by mass spectrometry using different ion sources. Two distributions could be identified in the mass spectra which related to two different polymer series (one with hydrogen and hydroxyl end-groups and the other with methyl and hydroxyl end-groups). The fragmentation behaviour of the protonated oligomers was studied in a quadrupole time-of-flight mass spectrometer (MS) with electrospray, atmospheric pressure chemical ionization and direct analysis in real time soft ionization techniques. Three product ion series were identified in the MS/MS spectra independently of the ion source used. Based on the results, a mechanism was proposed for the dissociation by means of the accurate mass of the product ions, pseudo MS(3) experiments and the energy dependence of the product ion intensity, i.e. breakdown curves. The survival yield method was used to highlight the correlation between the size of the oligomers and the laboratory frame collision energy.

The method of distance measurement and torso length influences the relationship of pulse wave velocity to cardiovascular mortality.

BACKGROUND: The method of estimating distance traveled by the pulse wave, used in the calculation of pulse wave velocity (PWV), is not standardized. Our objective was to assess whether different methods of distance measurement influenced the association of PWV to cardiovascular mortality in hemodialysis (HD) patients. METHODS: Ninety-eight chronic HD patients had their PWV measured using three methods for distance estimation; PWV1: suprasternal notch-to-femoral site minus suprasternal notch-to-carotid site, PWV2: carotid-to-femoral site, PWV3: carotid-to-femoral site minus suprasternal notch-to-carotid site. Carotid-to-femoral distance was used to approximate torso length. Patients were followed for a median of 30 months and the association of PWV and cardiovascular mortality was assessed using survival analysis before and after stratification for torso length. RESULTS: The three methods resulted in significantly different PWV values. During follow-up 50 patients died, 32 of cardiovascular causes. In log-rank tests, only tertiles of PWV1 was significantly related to outcome (P values 0.017, 0.257, 0.137, for PWV1, PWV2, and PWV3, respectively). In adjusted Cox, proportional hazards regression only PWV1 was related to cardiovascular mortality. In stratified analysis, however, among patients with below median torso length all PWV values were related to outcome, whereas in patients with above median torso length none of the PWV methods resulted in significant relationship to outcome. CONCLUSIONS: PWV calculated using suprasternal notch-to-femoral distance minus suprasternal notch-to-carotid distance provides the strongest relationship to cardiovascular mortality. Longer torso weakens the predictive value of PWV, possibly due to more tortuosity of the aorta hence, more error introduced when using surface tape measurements.

[Significance of arterial stiffness parameters for predicting cardiovascular mortality in hemodialysis patients: a prospective cohort study]. / A különbözô érfali tágulékonysági paraméterek jelentôsége a cardiovascularis mortalitás elôrejelzésében hemodializált betegek között: prospektív kohorszvizsgálat.

UNLABELLED: Previous studies demonstrated that different parameters of arterial stiffness are related to cardiovascular mortality in hemodialysis patients. The relative prognostic value of these parameters has not previously been evaluated in one cohort. PATIENTS AND METHODS: Carotid-femoral pulse wave velocity, carotid augmentation index, carotid pulse pressure and carotid-brachial pulse pressure amplification were measured in 98 patients before and after hemodialysis. Patients were followed for a median of 29 months (1-34) and the association of these parameters with cardiovascular mortality was assessed using log-rank tests and Cox proportional hazards regressions. RESULTS: During follow-up, 40 patients died (mortality rate 20.7/100 patient-year), of which 25 died of cardiovascular causes. Increasing pre- and postdialysis pulse wave velocity tertiles and decreasing predialysis pulse pressure amplification tertiles were significantly related to cardiovascular mortality (p-values are 0.012 and 0.011 for pre- and postdialysis pulse wave velocity, and <0.001 and 0,321 for pre- and postdialysis pulse pressure amplification, respectively). Neither the carotid augmentation index nor carotid pulse pressure was related to cardiovascular mortality. In the Cox-regression, the adjusted hazard ratios for 1 m/s higher pre- and postdialysis pulse wave velocity were 1.24 (1.07-1.44) and 1.17 (1.06-1.28), respectively. The hazard ratio for 10% lower predialysis pulse pressure amplification was 1.41 (1.03-1.92). When included in the same model, both predialysis pulse wave velocity and pulse pressure amplification remained significantly associated with cardiovascular mortality (relative risk: 1.23 [1.07-1.42] and 1.39 [1.02-1.89]). CONCLUSION: Among different stiffness parameters, pulse wave velocity is consistently related to cardiovascular mortality, irrespective of the timing of measurement. Predialysis pulse pressure amplification seems to provide additional prognostic information.

Arterial stiffness in hemodialysis: which parameter to measure to predict cardiovascular mortality?

In previous studies, different parameters of arterial stiffness were related to cardiovascular mortality in hemodialysis patients, but their relative prognostic value has not previously been evaluated in 1 cohort. Carotid-femoral pulse wave velocity (PWV), the carotid augmentation index, carotid pulse pressure (CPP) and carotid-brachial pulse pressure amplification (AMP) were measured in 98 patients before and after hemodialysis. Patients were followed for a median of 29 months (1-34) and the association of these parameters with cardiovascular mortality were assessed using log-rank tests and Cox proportional hazards regressions. During follow-up, 25 patients died of cardiovascular causes. Increasing pre- and postdialysis PWV tertiles and decreasing predialysis AMP tertiles were significantly related to cardiovascular mortality (p = 0.012 and 0.011 for PWV, respectively; < 0.001 for AMP). Neither the carotid augmentation index nor carotid pulse pressure were related to cardiovascular mortality. The adjusted hazard ratios for 1 m/s higher pre- and postdialysis PWV were 1.24 (1.07-1.44) and 1.17 (1.06-1.28), respectively. The hazard ratio for 10% lower predialysis AMP was 1.41 (1.03-1.92). When included in the same model, both predialysis PWV and AMP remained significantly associated with cardiovascular mortality. Among different stiffness parameters, PWV is consistently related to cardiovascular mortality, irrespective of the timing of measurement. Predialysis AMP seems to provide additional prognostic information.

Association between restless legs syndrome and depression in patients with chronic kidney disease.

Restless legs syndrome (RLS) is reportedly associated with depression. This association may be mediated by both sleep-dependent and sleep-independent mechanisms. Here we analyze the association between RLS and depressive symptoms in patients with chronic kidney disease (CKD). We also assessed whether the relationship is independent of insomnia. In a cross-sectional study, socio-demographic parameters, laboratory data, and medical history were collected from 788 kidney transplant patients and 161 dialyzed patients. Insomnia, depression, and the presence of RLS symptoms were assessed with standard questionnaires. Patients with probable RLS had a higher prevalence of depressive symptoms than those without RLS (56% vs. 22% with vs. without RLS, respectively; P<.001). Patients presenting RLS symptoms had higher Athens Insomnia Scale (AIS) scores than patients without RLS [median AIS score (interquartile range): 7 (6) vs. 3 (4) with vs. without RLS, respectively; P<.001]. The AIS score correlated with the CES-D score (Spearman's rho=0.54, P<.001). In multivariate analysis, the presence of RLS symptoms was independently associated with depressive symptoms (OR=3.96, 95% CI 2.21-7.1, P<.001). This relationship remained significant even after including insomnia in the model (OR=2.9, CI 1.55-5.43, P<.001). The presence of RLS symptoms is associated with depression in patients with CKD. This relationship remained significant even after accounting for insomnia. Sleep-independent mechanisms may also contribute to the association between RLS and depression in patients with CKD.

Successful management of pregnancy with nephrotic syndrome due to preexisting membranous glomerulonephritis: a case report.

The influence of membranous glomerulonephritis (MGN) on maternal and fetal outcome is controversial, as is the effect of pregnancy on the course of preexisting nephrotic syndrome. We report a case of successful management of a pregnancy with preexisting severe nephrotic syndrome due to biopsy-proven primary MGN. Our patient became pregnant in a non-compliance period, discontinued the nephrological follow-up program and her kidney disease decompensated. From the 22nd gestational week the patient was treated with intermittent pulses of methylprednisolone (250 mg i.v.) and a maintenance dose of 32-64 mg/day orally, along with azathioprine 100 mg/day. She also received antihypertensive, diuretic, and anticoagulant therapy, and supplementation with fresh frozen plasma and albumin. In the 33rd gestational week a cesarean section was performed due to deteriorating creatinine clearance, low serum total protein levels, increasing edema and progression of intrauterine growth retardation of the fetus. Three months after delivery, the patient's renal disease went into complete remission. To our knowledge, this is the first report of using azathioprine during pregnancy with severe nephrotic syndrome due to primary MGN.

Modulation of adenosine-induced responses in the guinea-pig trachea during long-term caffeine treatment: possible role of epithelium.

The responses to adenosine were studied on isolated, methacholine-precontracted tracheal strips of guinea pigs in the course of long-term caffeine or solvent treatment. Guinea pigs were fed caffeine for 10 weeks (average serum caffeine concentration: 39.1 +/- 3.9 microM). In epithelium-intact tracheal preparations (EITPs), sensititization to adenosine-induced relaxation (AIR) developed. It attained a maximum in week 1 of caffeine treatment, and then its level diminished and disappeared completely by weeks 4 - 6. In epithelium-denuded tracheal preparations (EDTPs), an increase in the sensitivity to adenosine was observed from week 1 to week 10 (a 4 - 6-fold reduction in EC50). Use of a coaxial bioassay system confirmed the role of epithelium in this process. The enhancement of the AIR of the EITPs was not modified by inhibitors of cyclooxygenase and lipoxygenase. Following depletion of the neuropeptides by acute capsaicin pretreatment, the AIR of the EITPs was strongly enhanced after caffeine treatment for 6 weeks. In chronically caffeine-treated EITPs, the inhibition of neutral endopeptidase led to dramatic reduction of the AIR. On the basis of the results by inhibiting nitric oxide synthase, it can be supposed that nitric oxide released from EITPs of long-lasting caffeine-treated animals operated as a constrictor agent. Our results show that chronic caffeine treatment gives rise to an initial sensitization to adenosine of the EITPs, this being followed by the development of a specific adaptive process in the epithelial cells, which counterbalances the increased tracheal sensitivity to adenosine.

Self-association of bis-(alpha,beta-D-glucopyranosyl)-polyisobutylene.

In this paper, we report the structure and apparent molecular weights of bis-(alpha,beta-D-glucopyranosyl)-polyisobutylene (Gluc-PIB-Gluc) aggregates in CDCl(3) by NMR spectroscopy. Analysis of DOSY (diffusion-ordered NMR spectroscopy) experiments of a solution of Gluc-PIB-Gluc showed the presence of aggregates that were corroborated with dynamic light scattering. The structure of the aggregates was also studied by correlation spectroscopy (COSY) and nuclear Overhauser effect spectroscopy (NOESY) experiments.