Polymorphisms in the promoter regions of MMP-2, MMP-3, MMP-9 and MMP-12 genes as determinants of aneurysmal coronary artery disease

OBJECTIVES: Our hypothesis was that functional polymorphisms in matrix metalloproteinase (MMP) genes may act as susceptibility factors for the development of coronary aneurysms (CAs). BACKGROUND: Different forms of remodeling have been described at the level of coronary arteries; CA, reported in 1% to 5% of patients with angiographic evidence of coronary artery disease (CAD), are one of them. Matrix metalloproteinases have been implicated in the pathogenesis of aneurysm development through increased proteolysis of extracellular matrix proteins. METHODS: We screened 3,862 patients who underwent coronary angiography and identified 113 patients with CAD with at least one CA (CA group); these patients were matched with 226 patients with CAD without CA (control group). The -1,306 C/T MMP-2, 5A/6A MMP-3, CA-repeat MMP-9 and -82 A/G MMP-12 polymorphisms were determined. RESULTS: The MMP-2, MMP-9 and MMP-12 polymorphisms were not associated with CA. By contrast, the 5A/5A genotype of MMP-3 was significantly more frequent in the CA group than in the control group (31% vs. 18%, p = 0.015); similarly, the MMP-3 5A allele was more frequent in the CA group (p = 0.009). Three variables were independently associated with CA: the MMP-3 5A/5A genotype (odds ratio [OR] = 2.23, 95% confidence interval [CI] [1.27 to 3.93]), a previous myocardial infarction (OR = 1.91, 95% CI [1.14 to 3.20]) and a history of aortic aneurysm (OR = 21.06, 95% CI [2.35 to 188]). CONCLUSIONS: The MMP-3 5A allele is associated with the occurrence of CA. Our results suggest that an increased proteolysis in the arterial wall may act as a susceptibility factor for the development of CA in patients with coronary atherosclerosis.     

A test for the correct specification of marginal structural models

Marginal structural models (MSMs) allow estimating the causal effect of a time-varying exposure on an outcome in the presence of time-dependent confounding. The parameters of MSMs can be estimated utilizing an inverse probability of treatment weight estimator under certain assumptions. One of these assumptions is that the proposed causal model relating the outcome to exposure history is correctly specified. However, in practice, the true model is unknown. We propose a test that employs the observed data to attempt validating the assumption that the model is correctly specified. The performance of the proposed test is investigated with a simulation study. We illustrate our approach by estimating the effect of repeated exposure to psychosocial stressors at work on ambulatory blood pressure in a large cohort of white-collar workers in Québec City, Canada. Code examples in SAS and R are provided to facilitate the implementation of the test.     

Contribution of arterial stiffness and stroke volume to peripheral pulse pressure in ICU patients: an arterial tonometry study

Objective Peripheral arterial pulse pressure is increasingly used to assess hemodynamic status. Our aim was to test the respective influence of arterial stiffness, stroke volume, peripheral resistance, and various hemodynamic and demographic variables on peripheral pulse pressure in critically ill patients. Design Prospective study. Setting Medical intensive care unit of a university hospital. Interventions None. Patients 67 sinus rhythm patients (mean age 57 ± 17 years) of whom 17 received vasoactive agents. Measurements and results The stroke volume was calculated by Doppler echocardiography. Radial pressures were calibrated from systolic and diastolic brachial cuff pressures. Central aortic pressure was estimated by radial applanation tonometry. The arterial compliance was estimated from the aortic pressure curve using the area method and the arterial stiffness was calculated as 1/compliance. The influences of age, body surface area, arterial stiffness, stroke volume, peripheral resistance, and time intervals on peripheral pulse pressure were tested using univariate and multivariate analyses. The mean arterial pressure ranged from 42 to 113 mmHg. Peripheral pulse pressure (59 ± 17 mmHg) was higher than aortic pulse pressure (40 ± 14 mmHg, p < 0.001). In patients aged ≥ 60 years whose mean arterial pressure was ≥ 80 mmHg, peripheral pulse pressure was related to arterial stiffness (r ² = 0.41) and to stroke volume (multiple r ² = 0.90). A similar but weaker relationship was observed in the overall population (multiple r ² = 0.52). Conclusions In critically ill patients, and especially in aged subjects with hemodynamic stability, peripheral pulse pressure mainly reflected the combined influences of arterial stiffness and stroke volume. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Epidermal‐like architecture obtained from equine keratinocytes in three‐dimensional cultures

Despite the high prevalence of skin conditions in the horse, there is a dearth of literature on the culture and biology of equine skin cells, and this is partially attributable to the lack of suitable in vitro skin models. The objective of this study was to develop a three‐dimensional (3D) culture system that would support the proliferation and differentiation of equine keratinocytes, similar to that observed in natural epidermis. Cell monolayers were obtained from explants of equine skin and serially passaged as highly pure keratinocyte populations (> 95% of cells), based on their expression of cytokeratins, including CK‐5 and CK‐14, which are associated in vivo with proliferating keratinocyte populations. Explant‐derived keratinocytes were seeded into Alvetex? 3D tissue scaffolds for 30 days under conditions that promote cell differentiation. Ultrastructural, immunohistochemical and biochemical analyses revealed that keratinocytes within scaffolds were able to proliferate and attain tissue polarity, including differentiation into basal and suprabasal layers. The basal layer contained distinct cuboidal cells with large nuclei and stained for proliferative markers such as CK‐5 and CK‐14. In contrast, the suprabasal layers consisted of cells with distinct polyhedral morphology, abundant cytoplasmic processes and desmosomes indicative of stratum spinosum and distinct flattened cornified cells that expressed involucrin, a marker of terminal differentiation. Thus, keratinocytes derived from primary equine skin explants were able to attain epidermal‐like architecture in culture. This novel system could provide a very useful tool for modelling skin diseases, drug testing/toxicity studies and, potentially, equine regenerative medicine. Copyright © 2016 John Wiley & Sons, Ltd.     

Gender Specificities in Risk Stratification After Myocardial Infarction

Objective: Although gender specificities of various risk factors have been well documented, risk stratification after myocardial infarction has never been compared in women and men. Methods: The power of left ventricular ejection fraction, heart rate variability, and mean RR interval computed from 24-hour Holter recordings, was compared in women and men for the prediction of cardiac mortality after an acute myocardial infarction. The study population consisted of 456 patients (108 women, 348 men) aged 50–75 years. Results: During a follow-up of 3 years, there were 41 cardiac deaths (13 women vs 28 men, P = NS). The positive predictive accuracy of left ventricular ejection fraction, heart rate variability, and mean RR interval at all sensitivity levels was higher in women than in men. For a 40% sensitivity, positive predictive accuracy of left ventricular ejection fraction was 46% in women and 16% in men (P < 0.05), positive predictive accuracy of mean RR interval was 90% in women and 28% in men (P < 0.05), and positive predictive accuracy of heart rate variability was 61% in women and 43% in men (P = NS). Mean RR interval had the highest positive predictive accuracy for cardiac mortality in women, but its superiority over heart rate variability was not statistically significant. In men, heart rate variability was the strongest predictor of mortality that was significantly more powerful than mean RR interval and left ventricular ejection fraction (P < 0.05). Conclusion: Increased 24-hour mean heart rate is the strongest predictor of cardiac mortality in women in whom it performs significantly better than in men. While in men, heart rate variability is a significantly better predictor of postinfarction cardiac mortality than 24-hour mean heart rate, this is not the case in women.     

Changes in myocardial electrical impedance in human heart graft rejection

BACKGROUND: Monitoring of post-transplant heart rejection is currently based on endomyocardial biopsy analysis. This study aimed to assess the effects of heart graft rejection on myocardial electrical impedance. METHODS AND RESULTS: Twenty-nine cardiac transplant patients and 9 controls underwent measurement of myocardial electrical impedance using a specifically designed amplifying system. The module and phase angle of myocardial impedance were measured. Histopathological rejection grading was performed according to ISHLT classification. Fifty impedance tests were performed in transplanted patients. Myocardial impedance (Z) was higher in controls than in transplanted patients (p<0.001) and followed a progressive decline at increasing current frequencies (p<0.001). Likewise, the phase angle of impedance in controls ranged from positive values at low frequencies to negative values at higher frequencies (from 2.5+/-0.9 degrees at 10 kHz to -3.8+/-2.1 degrees at 300 kHz, p<0.001). Rejection was associated with a significant decrease in myocardial impedance (Z) (15+/-6.6 Omega in grade 0, 13+/-6.0 Omega in grade 1A, and 3.3+/-0.9 Omega in grade 3A at 10 kHz, p<0.003). CONCLUSIONS: Mild degrees of cardiac graft rejection are associated with significant changes in myocardial electrical impedance in transplant patients. Further clinical investigation is warranted to assess the potential of cardiac impedance to detect heart graft rejection.     

Urokinase-type plasminogen activator regulates neurodegeneration and neurogenesis but not vascular changes in the mouse hippocampus after status epilepticus

Expression of urokinase-type plasminogen activator (uPA) is increased after brain injury, suggesting that, like in cancer tissue, uPA plays roles in brain remodeling. Here we injured brain with intrahippocampal kainic acid (KA) injection in adult Wt and uPA?/? mice. At 20 days post-injury, uPA?/? mice had more severe loss of contralateral pyramidal (p < 0.05) and hilar neurons (p < 0.05) than Wt mice. The number of doublecortin (DCX)-positive newly born neurons was also reduced in uPA?/? mice as compared to Wt (p < 0.01). No difference was observed in granule cell dispersion or distribution of DCX-positive neurons in the dentate gyrus. uPA deficiency did not affect the total length of hippocampal blood vessels or vessel density. No differences were observed in the severity of status epilepticus or consequent epilepsy between the genotypes. These data indicate that uPA deficiency can unfavorably modulate both delayed neurodegeneration and neurogenesis but has little effect on post-injury neuronal migration and vascular density. Our results favor the idea that elevated uPA during the post-injury phase is neuroprotective.