Rapid reduction of intracellular glutathione in human bronchial epithelial cells exposed to occupational levels of toluene diisocyanate.

Toluene diisocyanate (TDI) is a recognized chemical asthmogen, yet the mechanism of this toxicity and the molecular reactions involved have not been elucidated. We have previously shown that TDI vapor forms adducts with the apical surface of the respiratory epithelium, and that it colocalizes with ciliary tubulin. In vitro, we have shown rapid reaction of TDI with glutathione (GSH) and transfer of the bisGS-TDI adduct to a sulfhydryl-containing major histocompatibility complex peptide. This study sought to determine if intracellular GSH is altered following exposure to TDI. We used the dye CellTracker Green (chloromethylfluorescein, CMFDA) for detection of glutathione. One-day and 6-day air-liquid cultures of human bronchoepithelial cells (HBE) were exposed to 20-100 ppb TDI vapor for 5, 15, or 30 min. Cells were subsequently imaged using a confocal microscope. Both 1- and 6-day cultures showed a decrease in intensity of the thiol staining as a function of the TDI exposure dose. Doses as low as 20 ppb, the current permissible exposure limit (PEL) to TDI, resulted in rapid (within 5 min) decreases in fluorescence. The decreased fluorescence was not due to cytotoxicity or decrease in either esterase or glutathione-S-transferase activity, enzymes necessary for activation of the fluorescence of CMFDA. The decrease in glutathione levels was verified using another fluorescent label, ThioGlo(TM) 1, and cell extracts. In addition, the mucus produced by 6-day air-liquid interface HBE cells in response to TDI exposure appeared to be protective, as HBE cells underlying mucus retained more fluorescence than did cells in the same cultures that were not covered with mucus. These results, along with previous data, strongly suggest that TDI enters pulmonary cells and reacts rapidly with intracellular GSH, and that this can occur at the current PEL of 20 ppb. This rapid reaction suggests the importance of cellular thiols in TDI-induced pulmonary disease.     

Cardiac Resynchronization Therapy in Patients With Heart Failure and Narrow QRS Complexes

Background

Cross correlation analysis (CCA) using tissue Doppler imaging has been shown to be associated with outcome after cardiac resynchronization therapy (CRT) in patients with heart failure (HF) with wide QRS. However, its significance in patients with narrow QRS treated with CRT is unknown.

Tissue Doppler imaging in coronary artery diseases and heart failure

Recent studies have explored the prognostic role of TDI-derived parameters in major cardiac diseases, such as coronary artery disease (CAD) and heart failure (HF). In these conditions, myocardial mitral annular systolic (S') and early diastolic (E') velocities have been shown to predict mortality or cardiovascular events. In heart failure non invasive assessment of LV diastolic pressure by transmitral to mitral annular early diastolic velocity ratio (E/E') is a strong prognosticator, especially when E/E' is > or =15. Moreover, other parameters derived by TDI, as cardiac time intervals and Myocardial Performance Index, might play a role in the prognostic stratification in CAD and HF. Recently, a threedimensional (3-D) TDI imaging modality, triplane TDI, has become available, and this allows calculation of 3-Dvolumes and LV ejection fraction. We present a brief update of TDI.     

Evaluation of Cytochrome P450 Selectivity for Hydralazine as an Aldehyde Oxidase Inhibitor for Reaction Phenotyping

Hydralazine has been reported as a selective mechanism-based inactivator of aldehyde oxidase (AO) and it is widely used in the pharmaceutical industry for reaction phenotyping to estimate fraction metabolized by AO and to identify AO substrates. In this study, however, hydralazine was found to inhibit CYP1A2, 2B6, 2D6, and 3A in human suspension hepatocytes under reaction phenotyping assay conditions, at concentrations that chemically knocked out most of the AO activities (≥50 μM). Furthermore, hydralazine is a time-dependent inhibitor of CYP1A2. Based on these findings, precautions need to be taken when using hydralazine as an AO inhibitor for in vitro studies because fraction metabolized by AO is likely to be overestimated and the likelihood of false positives in identifying AO substrates increases.     

Ochratoxin A in cereal-derived products in Turkey: Occurrence and exposure assessment

Eighty-three samples of cereal-derived products including 24 breakfast cereals, 24 cereal-based baby foods and 35 beers purchased from supermarkets and small shops of Adana in Turkey were analysed for the presence of ochratoxin A (OTA) using immunoaffinity column (IAC) clean-up and high performance liquid chromatography with fluorescence detection (HPLC–FD). The average OTA recoveries from spiked breakfast cereal and cereal-based baby food and beers samples were in the range of 79.33–83.86%, 72.93–80.34% and 76.47–83.11%, respectively. The relative standard deviations (RSD_r) of recoveries for breakfast cereals, cereal-based baby foods and beers were 1.1–3.39%, 2.56–8.37% and 5.73–13.61%, respectively.     

Improved coronary artery blood flow following the correction of systolic dyssynchrony with cardiac resynchronization therapy

Background

Coronary blood flow (CBF) is improved by cardiac resynchronization therapy (CRT) and impaired by right ventricular apical (RVA) pacing in patients with heart failure. However, the underlying mechanism remains unclear.

Methods

Twenty-nine non-ischemic heart failure patients who responded to CRT underwent transthoracic echocardiography examination including both left anterior descending (LAD) CBF and tissue Doppler imaging in 3 pacing modes: intrinsic conduction, RVA pacing and biventricular (BiV) pacing. LAD velocity-temporal integral (LAD-VTI) and duration were measured. Systolic dyssynchrony was assessed with the standard deviation of a 12-left ventricular segmental model (Ts-SD).

Results

BiV pacing improved while RVA pacing reduced CBF compared to intrinsic conduction (all p < 0.05). Both Ts-SD and ventricular septal velocity deteriorated during RVA pacing but improved during BiV pacing (all p < 0.05). When systolic dyssynchrony was induced, lower LAD-VTI (9.5 ± 3.4 versus 12.7 ± 5.1 cm, p = 0.001) and shorter LAD diastolic duration (483 ± 92 versus 542 ± 106 ms, p = 0.010) were detected than synchronous status. Systolic dyssynchrony was inversely related to septal velocity (r = − 0.41), p<0.001 and LAD-VTI (r = − 0.30, p = 0.007), with the latter found to be moderately correlated to septal velocity (r = 0.30, p = 0.007).

Conclusion

Regional LAD flow was improved in patients subjected to BiV but worsened in those treated with RVA pacing in non-ischemic heart failure CRT responders. Systolic dyssynchrony was more commonly observed in patients subjected to RVA pacing. Reduction of septal velocity with dyssynchrony may directly lead to reduced LAD flow. Improvement of septal velocity by CRT and hence LAD flow may be an important mechanism in determining the response to CRT.     

Super‐resolution track‐density imaging of thalamic substructures: Comparison with high‐resolution anatomical magnetic resonance imaging at 7.0T

The thalamus is one of the most important brain structures, with strong connections between subcortical and cortical areas of the brain. Most of the incoming information to the cortex passes through the thalamus. Accurate identification of substructures of the thalamus is therefore of great importance for the understanding of human brain connectivity. Direct visualization of thalamic substructures, however, is not easily achieved with currently available magnetic resonance imaging (MRI), including ultra‐high field MRI such as 7.0T, mainly due to the limited contrast between the relevant structures. Recently, improvements in ultra‐high field 7.0T MRI have opened the possibility of observing thalamic substructures by well‐adjusted high‐resolution T₁‐weighted imaging. Moreover, the recently developed super‐resolution track‐density imaging (TDI) technique, based on results from whole‐brain fiber‐tracking, produces images with sub‐millimeter resolution. These two methods enable us to show markedly improved anatomical detail of the substructures of the thalamus, including their detailed locations and directionality. In this study, we demonstrate the role of TDI for the visualization of the substructures of the thalamic nuclei, and relate these images to T₁‐weighted imaging at 7.0T MRI. Hum Brain Mapp 34:2538–2548, 2013. © 2012 Wiley Periodicals, Inc.