Molecular plasticity of retinal ganglion cells after partial optic nerve injury

In the past few years we established the partial crush of the optic nerve as an in vivo model system for the study of signaling pathways involved in molecular plasticity after axonal injury. The simplicity of this model at the cellular level allows decisive questions to be anwsered whilst functional aspects of visual information processing can be studied in parallel. A major advantage of a partial optic nerve crush model is the opportunity to directly compare different cell populations: (i) the rapidly degenerating retinal ganglion cells (RGC), (ii) the axotomized RGC population that eventually dies over the period of the next few weeks, (iii) the axotomized RGC population surviving for a long time in the retina without an axon and (iv) the surviving RGC population that maintains axonal connections to their brain targets. Thus, differential aspects of post-lesion plasticity can be analyzed. Using this axonal injury model we investigated the expression of immediate early genes, glutamate receptors, and other differentially expressed genes that we identified with a combined subtractive hybridization and suppression polymerase chain reaction (PCR) screen. Moreover, we characterized time course of cell death, the astroglia response of the retina and optic nerve as well as the topography of anterograde and retrograde axonal transport.     

Tumor-infiltrating T cells with similar antigen binding sites in different mucosa-associated lymphoid tissue type B cell lymphomas

B cell lymphomas of mucosa-associated lymphoid tissue (MALT)-type arise on the background of chronic inflammation due to either autoimmunity or infection at various sites of the organism. Characteristically the tumor-infiltrating T cells found in large numbers in MALT-type lymphomas are predominantly of CD4(+) phenotype and may have impact on tumor pathogenesis. To assess whether the chromosomal translocation t(11;18)(q21; q21) that is specific for at least a subset of MALT-type lymphomas may have an impact on tumor environment, we investigated the antigen binding sites of tumor-infiltrating T cells from one t(11;18)-positive tumor of the thyroid and one t(11;18)-negative tumor of the stomach. MHC Allelotyping was performed and revealed common alleles in HLA-DR and HLA-DQ loci. Cloning and sequencing of the complementary determining region 3 in V(beta2) chains of T cell receptors demonstrated the use of identical J(beta)segments in both patients, suggestive for recognition of the same antigen. We therefore suggest that tumor-infiltrating CD4(+) T cells are tightly integrated in MALT-type lymphoma immunoarchitecture irrespective of genetic background and localization of the tumors.     

Prediction of coronary artery disease by a systemic atherosclerosis score index derived from whole-body MR angiography

Background

Whole-body magnetic resonance angiography (WB-MRA) has shown its potential for the non-invasive assessment of nearly the entire arterial vasculature within one examination. Since the presence of extra-cardiac atherosclerosis is associated with an increased risk of coronary events, our goal was to establish the relationship between WB-MRA findings, including a systemic atherosclerosis score index, and the presence of significant coronary artery disease (CAD).

Methods

WB-MRA was performed on a 1.5T scanner in 50 patients scheduled to undergo elective cardiac catheterization for suspected CAD. In each patient, 40 extra-cardiac vessel segments were evaluated and assigned scores according to their luminal narrowing. The atherosclerosis score index (ASI) was generated as the ratio of summed scores to analyzable segments.

Results

ASI was higher in patients with significant (> 50% stenosis) CAD (n = 27) vs. patients without CAD (n = 22; 1.56 vs. 1.28, p = 0.004). ASI correlated with PROCAM (R = 0.57, p < 0.001) and Framingham (R = 0.36, p = 0.01) risk scores as estimates of the 10-year risk of coronary events. A ROC derived ASI of > 1.54 predicted significant CAD with a sensitivity of 59%, specificity of 86% and a positive predictive value of 84%. Logistic regression revealed ASI > 1.54 as the strongest independent predictor for CAD with a 11-fold increase in likelihood to suffer from significant coronary disease. On the contrary, while 15/27 (55%) of patients with CAD exhibited at least one extra-cardiac stenosis > 50%, only 3/22 (14%) of those patients without CAD did (p = 0.003). The likelihood for an extra-cardiac stenosis when CAD is present differed between vascular territories and ranged from 15% for a carotid stenosis to 44% for a stenosis in the lower extremities.

Conclusion

This study provides important new evidence for the close association of extra-cardiac and coronary atherosclerosis. The novel findings that a WB-MRA derived systemic atherosclerosis score index is not only associated with established cardiovascular risk scores but is also predictive of significant CAD suggest its potential prognostic implications and underline the importance to screen for coronary disease in patients with extra-cardiac manifestations of atherosclerosis.     

Endothelial dysfunction and infarct-size relate to impaired EDHF response in rat experimental chronic heart failure

BACKGROUND: The rat coronary ligation model of chronic heart failure has been extensively used to investigate its pathophysiology including the role of endothelial dysfunction. Inconsistent results have been obtained concerning the role of endothelial dilative mediators nitric oxide (NO) and endothelium-derived hyperpolarizing factor (EDHF). AIMS: Our aim was to investigate involvement of NO and EDHF in aortic endothelial dysfunction in this model and the influence of individual infarct sizes. Furthermore, we investigated whether it is justified to regard rats that failed to develop large infarct sizes as SHAM controls. METHODS: We performed coronary ligations and SHAM operations and studied acetylcholine (ACh)-induced relaxations and underlying endothelial mediators in isolated aortic rings 12 weeks after infarction. By then, cardiac and hemodynamic parameters were deteriorated in animals with large myocardial infarctions (large-MI, 35+/-3%), but not those with small myocardial infarctions (small-MI, 5+/-2%). RESULTS: Large-MI showed decreased ACh-induced relaxation compared to SHAM due to decreased contribution of EDHF which was inversely correlated with individual infarct-size. Interestingly, small-MI showed significantly increased ACh-induced relaxation compared to SHAM due to increased NO contribution. CONCLUSIONS: Our results suggest that impaired aortic endothelial dilatory function in large-MI is mainly due to an impaired EDHF response and strongly depends on individual infarct-size. In addition, endothelium-dependent relaxation of small-MI rats differed from SHAM, indicating that both groups may not be pooled to serve as controls. These results emphasize the importance of infarct-size and choice of the control group, and may explain inconsistencies in previous studies.     

Increased expression of cardiac angiotensin II type 1 (AT(1)) receptors decreases myocardial microvessel density after experimental myocardial infarction

OBJECTIVE: To study the effects of increased levels of myocardial angiotensin II type 1 (AT(1)) receptor on microvascular growth following myocardial infarction (MI). METHODS: MI was created in transgenic rats (TGR) with a cardioselective overexpression of the AT(1) receptor. We used Sprague-Dawley (SD) rats as controls. Some of the rats were treated with the selective AT(1) receptor blocker losartan (Los). Rats were sacrificed after 3 weeks. RESULTS: MI caused left ventricular (LV) hypertrophy and LV dysfunction in both SD and TGR, which was prevented by AT(1) receptor blockade. Furthermore, MI decreased microvessel density in the non-infarcted myocardium (SD MI: 1653+/-37/mm(2), P<0.01 vs. sham-operated controls), however, microvessel density decreased significantly more in TGR with MI (1298+/-33/mm(2), P<0.01 vs. SD MI). AT(1) receptor blockade restored microvessel density (SD MI Los: 2046+/-195/mm(2); TGR MI Los: 1742+/-47/mm(2); P<0.01 vs. untreated). The differences in microvessel density were still present after correction for LV hypertrophy. The increase in microvessel density after AT(1) receptor blockade was not accompanied by increased myocardial vascular endothelial growth factor (VEGF) levels. Microvessel density correlated with parameters of myocardial stretch, such as LV end-diastolic pressure (-0.681, P<0.001) and N-ANP (-0.424, P=0.01). CONCLUSIONS: Microvessel density after MI is decreased when the AT(1) receptor is overexpressed, and this is amenable to AT(1) receptor blockade. This suggests that efficacy of AT(1) receptor blockers post-MI may not only be due to attenuation of LV remodeling, but also to a stimulatory effect on angiogenesis.     

Functional reconstitution of COPI coat assembly and disassembly using chemically defined components

Coat protein I (COPI)-coated transport vesicles mediate protein and lipid transport in the early secretory pathway. The basic machinery required for the formation of these transport intermediates has been elucidated based on the reconstitution of COPI-coated vesicle formation from chemically defined liposomes. In this experimental system, the coat components coatomer and GTP-bound ADP-ribosylation factor (ARF), as well as p23 as a membrane-bound receptor for COPI coat proteins, were shown to be both necessary and sufficient to promote COPI-coated vesicle formation. Based on biochemical and ultrastructural analyses, we now demonstrate that the catalytic domain of ARF-GTPase-activating protein (GAP) alone is sufficient to initiate uncoating of liposome-derived COPI-coated vesicles. By contrast, ARF-GAP activity is not required for COPI coat assembly and, therefore, does not seem to represent an essential coat component of COPI vesicles as suggested recently [Yang, J. S., Lee, S. Y., Gao, M., Bourgoin, S., Randazzo, P. A., et al. (2002) J. Cell Biol. 159, 69-78]. Thus, a complete round of COPI coat assembly and disassembly has been reconstituted with purified components defining the core machinery of COPI vesicle biogenesis.     

Acute Administration of Angiotensin Converting Enzyme Inhibitors in Thrombolysed Myocardial Infarction Patients Is Associated with a Decreased Incidence of Heart Failure,but an Increased Re-Infarction Risk

Introduction: Ventricular remodeling starts very early after the onset of an acute myocardial infarction (AMI), and can be prevented by ACE-inhibitors. However, very limited data are available on the effect of acute (< 9 hours) treatment with angiotensin converting enzyme (ACE) inhibitors after an AMI on mortality, heart failure and re-infarction. The aim of the present study was to evaluate the effects of acute ACE-inhibitor treatment.Methods: We performed a pooled analysis of three very similar randomized, placebo-controlled multi-center trials. In 845 thrombolysed patients with mainly first anterior MI, patients were randomised to acute ACE-inhibitor treatment (< 9 hours after MI) or placebo.Results: After acute ACE-inhibitor treatment we observed similar 3-months mortality rates, and a significant reduction in the incidence of 3-months heart failure (26.1 vs. 19.3%; RR 0.67; 95% CI 0.45–1.0) as compared to placebo. In contrast, acute ACE-inhibitor treatment was associated with a significant 2.0 times increased 3-months re-infarction risk (7.0 vs. 3.6%; RR 2.0; 95% CI 1.1 to 3.8). Subgroup-analysis showed that patients with small infarct sizes treated with acute ACE-inhibitor (peak CPK < 1000 IU) had a 7.6 times higher re-infarction risk (95% CI 1.7 to 33) than patients with small infarctions treated with placebo.Conclusions: Acute ACE-inhibitor treatment in thrombolysed patients with mainly first anterior AMI resulted in a reduction of heart failure and similar mortality, but an increase in re-infarction rates, especially in patients with small infarct sizes. These results warrant caution for the very early use of ACE-inhibitors in smaller infarctions, although this needs to be confirmed in a larger prospective randomised clinical trial.