A miniaturized culture platform for control of the metabolic environment.

The heart is a metabolic "omnivore" and adjusts its energy source depending on the circulating metabolites. Human cardiac organoids, a three-dimensional in vitro model of the heart wall, are a useful tool to study cardiac physiology and pathology. However, cardiac tissue naturally experiences shear stress and nutrient fluctuations via blood flow in vivo, whilst in vitro models are conventionally cultivated in a static medium. This necessitates the regular refreshing of culture media, which creates acute cellular disturbances and large metabolic fluxes. To culture human cardiac organoids in a more physiological manner, we have developed a perfused bioreactor for cultures in a 96-well plate format. The designed bioreactor is easy to fabricate using a common culture plate and a 3D printer. Its open system allows for the use of traditional molecular biology techniques, prevents flow blockage issues, and provides easy access for sampling and cell assays. We hypothesized that a perfused culture would create more stable environment improving cardiac function and maturation. We found that lactate is rapidly produced by human cardiac organoids, resulting in large fluctuations in this metabolite under static culture. Despite this, neither medium perfusion in bioreactor culture nor lactate supplementation improved cardiac function or maturation. In fact, RNA sequencing revealed little change across the transcriptome. This demonstrates that cardiac organoids are robust in response to fluctuating environmental conditions under normal physiological conditions. Together, we provide a framework for establishing an easily accessible perfusion system that can be adapted to a range of miniaturized cell culture systems.

Continuous shear wave measurements for dynamic cardiac stiffness evaluation in pigs.

Ultrasound-based shear wave elastography is a promising technique to non-invasively assess the dynamic stiffness variations of the heart. The technique is based on tracking the propagation of acoustically induced shear waves in the myocardium of which the propagation speed is linked to tissue stiffness. This measurement is repeated multiple times across the cardiac cycle to assess the natural variations in wave propagation speed. The interpretation of these measurements remains however complex, as factors such as loading and contractility affect wave propagation. We therefore applied transthoracic shear wave elastography in 13 pigs to investigate the dependencies of wave speed on pressure-volume derived indices of loading, myocardial stiffness, and contractility, while altering loading and inducing myocardial ischemia/reperfusion injury. Our results show that diastolic wave speed correlates to a pressure-volume derived index of operational myocardial stiffness (R = 0.75, p < 0.001), suggesting that both loading and intrinsic properties can affect diastolic wave speed. Additionally, the wave speed ratio, i.e. the ratio of systolic and diastolic speed, correlates to a pressure-volume derived index of contractility, i.e. preload-recruitable stroke work (R = 0.67, p < 0.001). Measuring wave speed ratio might thus provide a non-invasive index of contractility during ischemia/reperfusion injury.

Impact of Loading and Myocardial Mechanical Properties on Natural Shear Waves: Comparison to Pressure-Volume Loops.

BACKGROUND: Shear wave elastography (SWE) has been proposed as a novel noninvasive method for the assessment of myocardial stiffness, a relevant determinant of diastolic function. It is based on tracking the propagation of shear waves, induced, for instance, by mitral valve closure (MVC), in the myocardium. The speed of propagation is directly related to myocardial stiffness, which is defined by the local slope of the nonlinear stress-strain relation. Therefore, the operating myocardial stiffness can be altered by both changes in loading and myocardial mechanical properties. OBJECTIVES: This study sought to evaluate the capability of SWE to quantify myocardial stiffness changes in vivo by varying loading and myocardial tissue properties and to compare SWE against pressure-volume loop analysis, a gold standard reference method. METHODS: In 15 pigs, conventional and high-frame rate echocardiographic data sets were acquired simultaneously with pressure-volume loop data after acutely changing preload and afterload and after inducting an ischemia/reperfusion (I/R) injury. RESULTS: Shear wave speed after MVC significantly increased by augmenting preload and afterload (3.2 ± 0.8 m/s vs 4.6 ± 1.2 m/s and 4.6 ± 1.0 m/s, respectively; P = 0.001). Preload reduction had no significant effect on shear wave speed compared to baseline (P = 0.118). I/R injury resulted in significantly higher shear wave speed after MVC (6.1 ± 1.2 m/s; P < 0.001). Shear wave speed after MVC had a strong correlation with the chamber stiffness constant ß (r = 0.63; P < 0.001) and operating chamber stiffness dP/dV before induction of an I/R injury (r = 0.78; P < 0.001) and after (r = 0.83; P < 0.001). CONCLUSIONS: Shear wave speed after MVC was influenced by both acute changes in loading and myocardial mechanical properties, reflecting changes in operating myocardial stiffness, and was strongly related to chamber stiffness, invasively derived by pressure-volume loop analysis. SWE provides a novel noninvasive method for the assessment of left ventricular myocardial properties.

Increasing the Mechanical Strength and Corrosion Resistance of Aluminum Alloy 7075 via Hydrostatic Extrusion and Aging.

The present study investigates the correlation between mechanical properties and resistance to corrosion of hydrostatically extruded aluminum alloy 7075. Supersaturated solid solutionized samples undergo a plastic deformation process, followed by both natural and artificial aging. Furthermore, two types of hydrostatic extrusion are applied to the samples: single-stepped and double-stepped. This process is shown to influence grain refinement and the precipitation process, resulting in changes in the electrochemical properties of the samples. Hydrostatic extrusion combined with aging is shown to cause an increase in mechanical strength ranging from 50 MPa to 135 MPa in comparison to coarse-grained sample subjected to T6 heat treatment. The highest value of tensile strength is obtained for a sample subjected to single-step hydrostatic extrusion followed by natural aging. This strength increase is caused by refinement of the microstructure, in addition to the small size and number of precipitates at the grain boundaries, which are coarsened by artificial aging. Hydrostatic extrusion is also shown to increase resistance to corrosion, with the T6-treated coarse-grained sample being most susceptible to corrosion attack.

The Influence of Heat Treatment on the Mechanical Properties and Corrosion Resistance of the Ultrafine-Grained AA7075 Obtained by Hydrostatic Extrusion.

In this paper, the corrosion resistance and mechanical properties of the 7075 aluminum alloy are studied. The alloy was deformed by hydrostatic extrusion and then aged both naturally and artificially. Results are compared with those of coarse-grained material subjected to T6 heat treatment. The aim of the research is to find the optimal correlation between the mechanical properties and the corrosion resistance of the alloy. To this end, static tensile tests with subsequent fractography, open circuit potential, and potentiodynamic polarization tests in 0.05 M NaCl were conducted. Obtained results show that a combination of precipitate hardening and a deformed microstructure leads to increased mechanical strength with high anisotropy due to the presence of fibrous grains. Plastic deformation increases susceptibility to corrosion due to the increased number of grain boundaries, which act as paths along that corrosion propagates. However, further artificial aging incurs a positive effect on corrosion resistance due to changes in the chemical composition of the matrix as a result of the precipitation process.

High-Frame-Rate Speckle Tracking for Echocardiographic Stress Testing.

Stress echocardiography helps to diagnose cardiac diseases that cannot easily be detected or do not even manifest at rest. In clinical practice, assessment of the stress test is usually performed visually and, therefore, in a qualitative and subjective way. Although speckle tracking echocardiography (STE) has been proposed for the quantification of function during stress, its time resolution is inadequate at high heart rates. Recently, high-frame-rate (HFR) imaging approaches have been proposed together with dedicated STE algorithms capable of handling small interframe displacements. The aim of this study was to determine if HFR STE is effective in assessing strain and strain rate parameters during echocardiographic stress testing. Specifically, stress echocardiography, at four different workload intensities, was performed in 25 healthy volunteers. At each stress level, HFR images from the apical four-chamber view were recorded using the ULA-OP 256 experimental scanner. Then, the myocardium was tracked with HFR STE, and strain and strain rate biomarkers were extracted to further analyze systolic and diastolic (early and late) peaks, as well as a short-lived isovolumic relaxation peak during stress testing. The global systolic strain response was monophasic, revealing a significant (p < 0.001) increase at low stress but then reaching a plateau. In contrast, all strain rate indices linearly increased (p < 0.001) with increasing stress level. These findings are in line with those reported using tissue Doppler imaging and, thus, indicate that HFR STE can be a useful tool in assessing cardiac function during stress echocardiography.

In Vivo Comparison of Multiline Transmission and Diverging Wave Imaging for High-Frame-Rate Speckle-Tracking Echocardiography.

High-frame-rate (HFR) speckle-tracking echocardiography (STE) assesses myocardial function by quantifying motion and deformation at high temporal resolution. Among the proposed HFR techniques, multiline transmission (MLT) and diverging wave (DW) imaging have been used in this context both being characterized by specific advantages and disadvantages. Therefore, in this article, we directly contrast both approaches in an in vivo setting while operating at the same frame rate (FR). First, images were recorded at baseline (resting condition) from healthy volunteers and patients. Next, additional acquisitions during stress echocardiography were performed on volunteers. Each scan was contoured and processed by a previously proposed 2-D HFR STE algorithm based on cross correlation. Then, strain curves and their end-systolic (ES) values were extracted for all myocardial segments for further statistical analysis. The baseline acquisitions did not reveal differences in estimated strain between the acquisition modes ( ); myocardial segments ( ); or an interaction between imaging mode and depth ( ). Similarly, during stress testing, no difference ( p = 0.7 ) was observed for the two scan sequences, stress levels or an interaction sequence-stress level ( p = 0.94 ). Overall, our findings show that MLT and DW compoundings give comparable HFR STE strain values and that the choice for using one method or the other may thus rather be based on other factors, for example, system requirements or computational cost.

Evaporation-Driven Flow in Micropillar Arrays: Transport Dynamics and Chemical Analysis under Varied Sample and Ambient Conditions.

Microfluidic flow in lab-on-a-chip devices is typically very sensitive to the variable physical properties of complex samples, e.g., biological fluids. Here, evaporation-driven fluid transport (transpiration) is achieved in a configuration that is insensitive to interfacial tension, salinity, and viscosity over a wide range. Micropillar arrays ("pillar cuvettes") were preloaded by wicking a known volatile fluid (water) and then adding a microliter sample of salt, surfactant, sugar, or saliva solution to the loading zone. As the preloaded fluid evaporates, the sample is reliably drawn from a reservoir through the pillar array at a rate defined by the evaporation of the preloaded fluid (typically nL/s). Including a reagent in the preloaded fluid allows photometric reactions to take place at the boundary between the two fluids. In this configuration, a photometric signal enhancement is observed and chemical analysis is independent of both humidity and temperature. The ability to reliably transport and sense an analyte in microliter volumes without concern over salt, surfactant, viscosity (in part), humidity, and temperature is a remarkable advantage for analytical purposes.

Shear Wave Elastography Using High-Frame-Rate Imaging in the Follow-Up of Heart Transplantation Recipients.

OBJECTIVES: The purpose of this study was to investigate whether propagation velocities of naturally occurring shear waves (SWs) at mitral valve closure (MVC) increase with the degree of diffuse myocardial injury (DMI) and with invasively determined LV filling pressures as a reflection of an increase in myocardial stiffness in heart transplantation (HTx) recipients. BACKGROUND: After orthotopic HTx, allografts undergo DMI that contributes to functional impairment, especially to increased passive myocardial stiffness, which is an important pathophysiological determinant of left ventricular (LV) diastolic dysfunction. Echocardiographic SW elastography is an emerging approach for measuring myocardial stiffness in vivo. Natural SWs occur after mechanical excitation of the myocardium, for example, after MVC, and their propagation velocity is directly related to myocardial stiffness, thus providing an opportunity to assess myocardial stiffness at end-diastole. METHODS: A total of 52 HTx recipients who underwent right heart catheterization (all) and cardiac magnetic resonance (CMR) (n = 23) during their annual check-up were prospectively enrolled. Echocardiographic SW elastography was performed in parasternal long axis views of the LV using an experimental scanner at 1,135 ± 270 frames per second. The degree of DMI was quantified with T1 mapping. RESULTS: SW velocity at MVC correlated best with native myocardial T1 values (r = 0.75; p < 0.0001) and was the best noninvasive parameter that correlated with pulmonary capillary wedge pressures (PCWP) (r = 0.54; p < 0.001). Standard echocardiographic parameters of LV diastolic function correlated poorly with both native T1 and PCWP values. CONCLUSIONS: End-diastolic SW propagation velocities, as measure of myocardial stiffness, showed a good correlation with CMR-defined diffuse myocardial injury and with invasively determined LV filling pressures in patients with HTx. Thus, these findings suggest that SW elastography has the potential to become a valuable noninvasive method for the assessment of diastolic myocardial properties in HTx recipients.

Investigation of Microwave Absorption Performance of CoFe2O4/NiFe2O4/Carbon Fiber Composite Coated with Polypyrrole in X-Band Frequency.

The current research reports the preparation of a microwave absorber containing CoFe2O4/NiFe2O4/Carbon fiber (H/S/CF) coated with polypyrrole polymer (PPy@H/S/CF) through sol-gel and in-situ polymerization processes. X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), vibrating sample magnetometer (VSM), and a vector network analyzer (VNA) are utilized to evaluate the features of the prepared composite. The microstructure analysis results revealed carbon fibers well decorated with submicron-size particles having hard/soft magnetic phases and thoroughly coated with polymer. The paraffin-based microwave absorber sample filled with 45 wt.% of PPy@H/S/CF has simultaneously both magnetic and dielectric losses in the 8.2-12.4 GHz frequency range. The absorber is used in a Salisbury screen configuration aiming at reducing the radar cross-section of objects. A minimum reflection loss of -55 dB at 10.6 GHz frequency with 5 GHz bandwidth is obtained for the sample with a 2 mm thickness. Different mechanisms, such as interfacial polarization, ferromagnetic resonance, and electron hopping, are the main factors for achieving such an appropriate microwave absorption. These results suggest that the PPy@H/S/CF composite is an ideal candidate for microwave absorption applications requiring high performance and low thickness.

A Novel 2-D Speckle Tracking Method for High-Frame-Rate Echocardiography.

Speckle tracking echocardiography (STE) is a clinical tool to noninvasively assess regional myocardial function through the quantification of regional motion and deformation. Even if the time resolution of STE can be improved by high-frame-rate (HFR) imaging, dedicated HFR STE algorithms have to be developed to detect very small interframe motions. Therefore, in this article, we propose a novel 2-D STE method, purposely developed for HFR echocardiography. The 2-D motion estimator consists of a two-step algorithm based on the 1-D cross correlations to separately estimate the axial and lateral displacements. The method was first optimized and validated on simulated data giving an accuracy of ~3.3% and ~10.5% for the axial and lateral estimates, respectively. Then, it was preliminarily tested in vivo on ten healthy volunteers showing its clinical applicability and feasibility. Moreover, the extracted clinical markers were in the same range as those reported in the literature. Also, the estimated peak global longitudinal strain was compared with that measured with a clinical scanner showing good correlation and negligible differences (-20.94% versus -20.31%, p -value = 0.44). In conclusion, a novel algorithm for STE was developed: the radio frequency (RF) signals were preferred for the axial motion estimation, while envelope data were preferred for the lateral motion. Furthermore, using 2-D kernels, even for 1-D cross correlation, makes the method less sensitive to noise.

Interplay of cardiac remodelling and myocardial stiffness in hypertensive heart disease: a shear wave imaging study using high-frame rate echocardiography.

AIMS: To determine myocardial stiffness by means of measuring the velocity of naturally occurring myocardial shear waves (SWs) at mitral valve closure (MVC) and investigate their changes with myocardial remodelling in patients with hypertensive heart disease. METHODS AND RESULTS: Thirty-three treated arterial hypertension (HT) patients with hypertrophic left ventricular (LV) remodelling (59 ± 14 years, 55% male) and 26 aged matched healthy controls (55±15 years, 77% male) were included. HT patients were further divided into a concentric remodelling (HT1) group (13 patients) and a concentric hypertrophy (HT2) group (20 patients). LV parasternal long-axis views were acquired with an experimental ultrasound scanner at 1266 ± 317 frames per seconds. The SW velocity induced by MVC was measured from myocardial acceleration maps. SW velocities differed significantly between HT patients and controls (5.83 ± 1.20 m/s vs. 4.04 ± 0.96 m/s; P < 0.001). In addition, the HT2 group had the highest SW velocities (P < 0.001), whereas values between controls and the HT1 group were comparable (P = 0.075). Significant positive correlations were found between SW velocity and LV remodelling (interventricular septum thickness: r = 0.786, P < 0.001; LV mass index: r = 0.761, P < 0.001). SW velocity normalized for wall stress indicated that myocardial stiffness in the HT2 group was twice as high as in controls (P < 0.001), whereas values of the HT1 group overlapped with the controls (P = 1.00). CONCLUSIONS: SW velocity as measure of myocardial stiffness is higher in HT patients compared with healthy controls, particularly in advanced hypertensive heart disease. Patients with concentric remodelling have still normal myocardial properties whereas patients with concentric hypertrophy show significant stiffening.

Velocities of Naturally Occurring Myocardial Shear Waves Increase With Age and in Cardiac Amyloidosis.

OBJECTIVES: This study sought to evaluate whether velocity of naturally occurring myocardial shear waves (SW) could relate to myocardial stiffness (MS) in vivo. BACKGROUND: Cardiac SW imaging has been proposed as a noninvasive tool to assess MS. SWs occur after mechanical excitation of the myocardium (e.g., mitral valve closure [MVC] and aortic valve closure [AVC]), and their propagation velocity is theoretically related to MS, thus providing an opportunity to assess stiffness at end-diastole (ED) and end-systole. However, given that SW propagation in vivo is complex, it remains unclear whether natural SW velocity effectively relates to MS. METHODS: This study prospectively enrolled 50 healthy volunteers (HV) (43.7 ± 17.1 years of age) and 18 patients with cardiac amyloidosis (CA) (68.0 ± 9.8 years of age). HV were divided into 3 age groups: group I, 20 to 39 years of age (n = 24); group II, 40 to 59 years of age (n = 11); and group III, 60 to 80 years of age (n = 15). Parasternal long-axis views were acquired using an experimental scanner. Tissue (Doppler) acceleration maps were extracted from an anatomical M-mode along the midline of the left ventricular septum. RESULTS: SW propagation velocity was significantly higher in CA patients than in HV after both MVC (3.54 ± 0.93 m/s vs. 6.33 ± 1.63 m/s, respectively; p < 0.001) and AVC (3.75 ± 0.76 m/s vs. 5.63 ± 1.13 m/s, respectively; p < 0.001). Similarly, SW propagation velocity differed significantly among age groups in HV, with a significantly higher value for group III than for group I, both occurring after MVC (p < 0.001) and AVC (p < 0.01). Moreover, SW propagation velocity after MVC was found to be significantly higher in patients with an increasing grade of diastolic dysfunction (p < 0.001). Finally, positive correlation was found between SW velocities after MVC and mitral inflow-to-mitral relaxation velocity ratio (E/E') (r = 0.74; p = 0.002). CONCLUSIONS: End-diastole SW velocities were significantly higher in patients with CA, patients with a higher grade of diastolic dysfunction, and elderly volunteers. These findings thus suggest that the speed of naturally induced SWs may be related to MS.

Natural Shear Wave Imaging in the Human Heart: Normal Values, Feasibility, and Reproducibility.

Left ventricular myocardial stiffness could offer superior quantification of cardiac systolic and diastolic function when compared to the current diagnostic tools. Shear wave elastography in combination with acoustic radiation force has been widely proposed to noninvasively assess tissue stiffness. Interestingly, shear waves can also result from intrinsic cardiac mechanical events (e.g., closure of valves) without the need for external excitation. However, it remains unknown whether these natural shear waves always occur, how reproducible they can be detected and what the normal range of shear wave propagation speed is. The present study, therefore, aimed at establishing the feasibility of detecting shear waves created after mitral valve closure (MVC) and aortic valve closure (AVC), the variability of the measurements, and at reporting the normal values of propagation velocity. Hereto, a group of 30 healthy volunteers was scanned with high-frame rate imaging (>1000 Hz) using an experimental ultrasound system transmitting a diverging wave sequence. Tissue Doppler velocity and acceleration were used to create septal color M-modes, on which the shear waves were tracked and their velocities measured. Overall, the methodology was capable of detecting the transient vibrations that spread throughout the intraventricular septum in response to the closure of the cardiac valves in 92% of the recordings. Reference velocities of 3.2±0.6 m/s at MVC and 3.5±0.6 m/s at AVC were obtained. Moreover, in order to show the diagnostic potential of this approach, two patients (one with cardiac amyloidosis and one undergoing a dobutamine stress echocardiography) were scanned with the same protocol and showed markedly higher propagation speeds: the former presented velocities of 6.6 and 5.6 m/s; the latter revealed normal propagation velocities at baseline, and largely increased during the dobutamine infusion (>15 m/s). Both cases showed values consistent with the expected changes in stiffness and cardiac loading conditions.

Actinometric and biodosimetric evaluation of UV-C dose delivery in annular, Taylor-Coutte and coiled tube continuous systems.

In this study, the evaluation of the performance of two thin-film UV-C reactors (annular and Taylor-Couette) and a coiled tube system is presented using actinometry and biodosimetry methods. The iodide/iodate actinometry method was found suitable for comparison of the efficiency of UV-C dose delivery of the UV-C continuous flow systems. Inactivation kinetics of Escherichia coli ATCC 8739 in quarter-strength Ringer's solution (absorption coefficient α254 nm ∼ 0 cm-1) at various flow conditions at Reynolds numbers in the range of 26 to 3000 showed a good correlation between the different reactor types. In high UV-C absorbing liquids, the inactivation efficiency increases due to the improved radial mixing. The inactivation performance of the Taylor-Couette system correlates to the annular reactor when no rotation force is applied. The residence time distributions showed the narrowest distribution with the coiled tube system at comparable flow rates. The results indicate that, despite the laminar flow conditions, the performance of the Taylor-Couette unit becomes equal to the turbulent flow conditions of the coiled tube reactor by rotation of the inner cylinder.

A Comparison of Antibacterial Activity of Selected Thyme (Thymus) Species by Means of the Dot Blot Test with Direct Bioautographic Detection.

Bioautography carried out with the aid of thin-layer chromatographic adsorbents can be used to assess antibacterial activity in samples of different origin. It can either be used as a simple and cost-effective detection method applied to a developed chromatogram, or to the dot blot test performed on a chromatographic plate, where total antibacterial activity of a sample is scrutinized. It was an aim of this study to compare antibacterial activity of 18 thyme (Thymus) specimens and species (originating from the same gardening plot and harvested in the same period of time) by means of a dot blot test with direct bioautography. A two-step extraction of herbal material was applied, and at step two the polar fraction of secondary metabolites was obtained under the earlier optimized extraction conditions [methanol-water (27+73, v/v), 130°C]. This fraction was then tested for its antibacterial activity against Bacillus subtilis bacteria. It was established that all investigated extracts exhibited antibacterial activity, yet distinct differences were perceived in the size of the bacterial growth inhibition zones among the compared thyme species. Based on the results obtained, T. citriodorus "golden dwarf" (sample No. 5) and T. marschallianus (sample No. 6) were selected as promising targets for further investigations and possible inclusion in a herbal pharmacopeia, which is an essential scientific novelty of this study.

Antioxidant Activity of Selected Thyme (Thymus L.) Species and Study of the Equivalence of Different Measuring Methodologies.

This study presents the results of comparative evaluation of the antioxidant activity of the phenolic fraction exhaustively extracted with aqueous methanol from 18 different thyme (Thymus L.) specimens and species. This evaluation is made with use of the same free radical source (DPPH• radical), three different free radical scavenging models (gallic acid, ascorbic acid, and Trolox), and three different measuring techniques (the dot blot test, UV-Vis spectrophotometry, and electron paramagnetic resonance spectroscopy, EPR). A comparison of the equivalence of these three different measuring techniques (performed with use of hierarchical clustering with Euclidean distance as a similarity measure and Ward's linkage) is particularly important in view of the fact that different laboratories use different antioxidant activity measuring techniques, which makes any interlaboratory comparison hardly possible. The results obtained confirm a semiquantitative equivalence among the three compared methodologies, and a proposal is made of a simple and cost-effective dot blot test that uses the DPPH• radical and provides differentiation of antioxidant activity of herbal matter comparable with the results of the UV-Vis spectrophotometry and EPR.

NADPH oxidase inhibitor, apocynin, improves renal glutathione status in Zucker diabetic fatty rats: a comparison with melatonin.

Apocynin (4'-hydroxy-3'-methoxyacetophenone) is the most commonly used NADPH oxidase (Nox) inhibitor. However, its application raises serious controversies, as the compound has been reported to reveal some prooxidative effects. The aim of this study was to elucidate apocynin action on glutathione, the main intracellular antioxidant, metabolism in kidneys of Zucker diabetic fatty (ZDF) rat, a well established model of diabetes type 2. Additionally, apocynin effects were compared with those of melatonin. The experiments were performed on five groups of animals: (1) untreated lean (?/+) ZDF rats, (2) ZDF ?/+ rats treated with apocynin (2 g/l) in drinking water, (3) untreated obese diabetic (fa/fa) ZDF rats, (4) ZDF fa/fa rats treated with apocynin (2 g/l) in drinking water, and (5) ZDF fa/fa rats treated with melatonin (20 mg/l) in drinking water. After 8weeks of the treatment, the following parameters were measured in kidneys: NADPH oxidase activity, the rate of hydroxyl free radicals (HFR) production, GSH and GSSG content and the activities of the enzymes of glutathione metabolism: γ-glutamylcysteine synthetase (GCS), glutathione reductase (GR) and glutathione peroxidase (GPx). Compared to ?/+ controls, ZDF fa/fa rats exhibited increased Nox activity, accelerated HFR generation and dramatically lowered GSH/GSSG ratio accompanied by increased GPx and diminished GCS activities. In case of diabetic animals, apocynin treatment resulted in attenuation of both Nox activity and HFR production, restoration of control GSH/GSSG ratio (due to both an increase in GSH and a decline in GSSG content), normalization of GPx activity and a slight increase in GCS activity. Similar observations were made upon melatonin application to ZDF fa/fa rats. Thus, it is concluded that, in the diabetic model studied, apocynin extends a beneficial effect on renal glutathione homeostasis. The mechanism of this phenomenon involves attenuation of glutathione peroxidase activity, which is overstimulated under conditions of oxidative stress accompanying diabetes.

Fingerprinting of the volatile fraction from selected thyme species by means of headspace gas chromatography with mass spectrometric detection.

The Thymus genus has a firm position in Mediterranean folk medicine and culinary tradition, and yet certain confusion is observed in its botanical taxonomy. Therefore, chemical analysis of secondary metabolites and selection of reliable chemotaxonomic markers can prove helpful. In this study, the volatile fraction derived from 20 different thyme specimens was analyzed by means of headspace GC with MS detection. From the obtained chromatographic fingerprints, the diversity of the volatile fraction originating from the different thyme species clearly emerged. Identification of volatile components was based on a software library of mass spectra. There was only one common component derived from all 20 investigated thyme species, i.e., ß-linalool. For three different Thymus species, a comparison was made of the performance of the headspace extraction and hydrodistillation. Characteristic differences in composition of the volatile fraction obtained with use of these two techniques were discussed. Finally, the obtained chromatographic fingerprints of the volatile fraction were analyzed by means of a chemometric approach (principal component analysis), in order to trace similarities in their chemical composition.