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1.
J Clin Med ; 11(7)2022 Mar 22.
Article in English | MEDLINE | ID: mdl-35407370

ABSTRACT

Recent randomized controlled multi-center trials JUPITER, CANTOS and COLCOT impressively demonstrated the effect of anti-inflammatory therapy on secondary prevention of cardiovascular events. These studies also rapidly re-vitalized the question of whether the C-reactive protein (CRP), the prototype human acute phase protein, is actively involved in atherosclerosis and its sequelae. Direct CRP inhibition may indeed improve the specificity and effectiveness of anti-inflammatory intervention. In the present paper, we report on the final results of our single-center C-reactive protein-Digoxin Observational Study (C-DOS). Methods and Results: Based on the experimental finding that cardiac glycosides potently inhibit hepatic CRP synthesis on the transcriptional level in vitro, 60 patients with decompensated heart failure, NYHA III−IV, severely reduced Left Ventricular Ejection Fraction (LVEF < 40%), and elevated CRP plasma levels were treated by either digoxin + conventional heart failure therapy (30 patients) or by conventional heart failure therapy alone (30 patients). Plasma CRP levels in both groups were assessed for 21 d. Plasma CRP levels on d1, d3 and d21 were compared by regression analysis. CRP levels d21−d1 significantly declined in both groups. Notably, comparative CRP reduction d21−d3 in digoxin versus the control group also revealed borderline significance (p = 0.051). Conclusions: This small observational trial provides the first piece of evidence that cardiac glycosides may inhibit CRP synthesis in humans. In case of further pharmacological developments, cardiac glycosides may emerge as lead compounds for chemical modification in order to improve the potency, selectivity and pharmacokinetics of CRP synthesis inhibition in cardiovascular disease.

2.
Cardiovasc Revasc Med ; 40: 144-149, 2022 07.
Article in English | MEDLINE | ID: mdl-34844868

ABSTRACT

BACKGROUND: Radial (RA) instead of femoral access (FA) for coronary interventions has become a European Society of Cardiology Class-IA guideline recommendation. But when the decision on the access site is left to the discretion of the operator, differences in adverse event rates mitigate. METHODS: We compared the 30-day outcome for RA and FA in all patients recruited for the observational German Austrian ABSORB Registry (GABI-R) in regard to all-cause mortality, stroke, myocardial infarction (MI), TIMI major bleedings (TMB) and quality of life (QoL). All patients were treated with a bioresorbable vascular scaffold. Access site was left to the discretion of the operator. RESULTS: In total, 3137 patients included by 92 centers received percutaneous coronary interventions (PCI) for acute MI in 51.5% and non-acute settings in 48.5%. RA was performed in 47.8% and had a higher median radiation exposure (3896 vs. 3082 cGycm2, p < 0.001). There was no difference in the amount of contrast used. There was also no difference in all-cause mortality (0.53% vs. 0.49%, p = 0.86), the combination of death, MI and stroke (1.87% vs. 1.83%, p = 0.94), but a trend towards more TMB (0.47% vs. 1.04%, p = 0.07) with FA. These outcomes were consistent across the subgroups of patients with ST-elevation MI, non-ST-elevation-ACS and stable coronary artery disease. Finally, QoL did not differ between RA and FA. CONCLUSIONS: In this contemporary GABI-R cohort, in which access site was left to the discretion of the operator, both access routes were safe and equal concerning QoL (ClinicalTrials.gov; NCT02066623).


Subject(s)
Myocardial Infarction , Percutaneous Coronary Intervention , Stroke , Austria , Femoral Artery , Hemorrhage/etiology , Humans , Myocardial Infarction/etiology , Percutaneous Coronary Intervention/adverse effects , Quality of Life , Radial Artery , Registries , Stroke/diagnosis , Stroke/etiology , Treatment Outcome
3.
Am J Case Rep ; 22: e932964, 2021 Aug 05.
Article in English | MEDLINE | ID: mdl-34351878

ABSTRACT

BACKGROUND High C-reactive protein (CRP) plasma levels in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection are associated with poor prognosis. CRP, by activating the classical complement pathway and interacting with macrophages via Fc gamma receptors, can cause pulmonary inflammation with subsequent fibrosis. Recently, we have reported first-in-man CRP apheresis in a "high-risk" COVID-19 patient. Treatment was unfortunately clinically unsuccessful. Here, we report on successful CRP apheresis treatment in a "lower-risk" COVID-19 patient with respiratory failure. CASE REPORT A 39-year-old male patient suffering from fatigue, dyspnea, and fever for 4 days was referred to us. The patient had to be intubated. Polymerase chain reaction (PCR) analysis of a throat smear revealed SARS-CoV-2 infection. Mutation analysis revealed the VOC B. 1.1.7 variant. CRP levels were 79.2 mg/L and increased to 161.63 mg/L. Procalcitonin (PCT) levels were continuously normal (<0.5 ng/ml). Antibiotic therapy was started to avoid bacterial superinfection. CRP apheresis was performed once via central venous access. CRP levels declined from a maximum of 161.63 mg/L to 32.58 mg/L. No apheresis-associated adverse effects were observed. Subsequently, CRP plasma levels declined day by day and normalized on day 5. The patient was extubated on day 5 and discharged from the Intensive Care Unit (ICU) on day 6. A second low CRP peak (maximum 22.41 mg/L) on day 7 remained clinically inapparent. The patient was discharged in good clinical condition with a CRP level of 6.94 mg/L on day 8. CONCLUSIONS SARS-CoV-2 infection can induce an uncontrolled CRP-mediated autoimmune response of ancient immunity. In this patient, the autoimmune response was potently and successfully suppressed by early selective CRP apheresis.


Subject(s)
Blood Component Removal , COVID-19 , Respiratory Insufficiency , Adult , C-Reactive Protein , Humans , Male , SARS-CoV-2
4.
Am J Case Rep ; 21: e925020, 2020 Jul 14.
Article in English | MEDLINE | ID: mdl-32661220

ABSTRACT

BACKGROUND C-reactive protein (CRP) plasma levels in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a novel viral disease, are surprisingly high. Pulmonary inflammation with subsequent fibrosis in SARS-CoV-2 infection is strongly accelerated. Recently, we have developed CRP apheresis to selectively remove CRP from human plasma. CRP may contribute to organ failure and pulmonary fibrosis in SARS-CoV-2 infection by CRP-mediated complement and macrophage activation. CASE REPORT A 72-year-old male patient at high risk was referred with dyspnea and fever. Polymerase chain reaction analysis of throat smear revealed SARS-CoV-2 infection. CRP levels were ~200 mg/L. Two days after admission, CRP apheresis using the selective CRP adsorber (PentraSorb® CRP) was started. CRP apheresis was performed via peripheral venous access on days 2, 3, 4, and 5. Following a 2-day interruption, it was done via central venous access on days 7 and 8. Three days after admission the patient was transferred to the intensive care unit and intubated due to respiratory failure. Plasma CRP levels decreased by ~50% with peripheral (processed blood plasma ≤6000 mL) and by ~75% with central venous access (processed blood plasma ≤8000 mL), respectively. No apheresis-associated side effects were observed. After the 2-day interruption in apheresis, CRP levels rapidly re-increased (>400 mg/L) and the patient developed laboratory signs of multi-organ failure. When CRP apheresis was restarted, CRP levels and creatinine kinases (CK/CK-MB) declined again. Serum creatinine remained constant. Unfortunately, the patient died of respiratory failure on day 9 after admission. CONCLUSIONS This is the first report on CRP apheresis in a SARS-CoV-2 patient. SARS-CoV-2 may cause multi-organ failure in part by inducing an excessive CRP-mediated autoimmune response of the ancient innate immune system.


Subject(s)
Betacoronavirus , Blood Component Removal/methods , C-Reactive Protein/metabolism , Coronavirus Infections/therapy , Multiple Organ Failure/therapy , Pneumonia, Viral/therapy , Aged , COVID-19 , Coronavirus Infections/blood , Coronavirus Infections/complications , Humans , Male , Multiple Organ Failure/blood , Multiple Organ Failure/etiology , Pandemics , Pneumonia, Viral/blood , Pneumonia, Viral/complications , SARS-CoV-2
5.
Int J Cardiol Heart Vasc ; 27: 100501, 2020 Apr.
Article in English | MEDLINE | ID: mdl-32258361

ABSTRACT

AIMS: The ABSORB bioresorbable vascular scaffold raised safety concerns due to higher rates of scaffold thrombosis (ScT) and adequate scaffold diameter and length for scaffold technology. Smaller scaffold diameter (SScD, 2.5 mm) was an infrequently quoted predictor of major adverse cardiac events (MACE). Therefore, we evaluated the impact of SScD compared to large scaffold diameter (LScD, ≥3 mm) of ≤18 mm device length on 2 year outcome in the all-comer real life GABI-R cohort. METHODS AND RESULTS: We compared patients with implanted LScD (1341 patients) vs. SScD (444 patients) of ≤18 mm device length. Patients with LScD more often presented with ST-elevation myocardial infarction (35.8% vs. 20.6%, p < 0.0001) and single-vessel disease (50.6% vs. 36.5% p < 0.0001). After a 24 months follow-up, there was no difference in regard of MACE (9.66% vs. 12.31%, p = 0.14) or definite/probable ST (2.47% vs. 2.82%, p = 0.71). Despite no difference in target lesion revascularisations (TLR) (5.81% vs. 7.71%, p = 0.18), there was a higher need for target vessel revascularisation (TVR) in the SScD-group (11.57% vs. 7.51%, p < 0.05). CONCLUSION: Compared to LScD, SScD of ≤18 mm device length demonstrated comparable safety in regard to MACE and ScT as well as efficacy in regard to TLR. Resorbable scaffold technology should not be restricted to large vessel diameters. CLINICAL TRIAL REGISTRATION: https://clinicaltrials.gov/ct2/show/NCT02066623.

6.
Case Rep Cardiol ; 2018: 4767105, 2018.
Article in English | MEDLINE | ID: mdl-30533227

ABSTRACT

C-reactive protein (CRP) may be causative in cardiovascular disease. As yet, no specific CRP inhibitor for human application has been described. A 69-year-old male was referred with ST segment elevation myocardial infarction (STEMI). Typical symptoms of chest pain started at 10.00 p.m. The patient was admitted to the hospital at 1.30 a.m. the next day. As ECG showed anterior wall myocardial infarction, the patient was immediately transferred to successful emergency angioplasty/drug-eluting- (DE-) stenting of the subtotally occluded left anterior descending artery. Consecutively, the hemodynamically stable patient was monitored at the chest pain unit. C-reactive protein (CRP) apheresis using the CRP adsorber (PentraSorb® CRP) within CAMI-1 trial was performed 34 h and 58 h after the onset of symptoms. In each apheresis session, 6000 ml plasma was treated via peripheral venous access. Plasma CRP levels decreased from 28.77 mg/l to 12.58 mg/l during the first apheresis session and from 24.17 mg/l to 11.55 mg/l during the second session, respectively. No side effects were observed. This is the first report of selective CRP apheresis in a man. The technology offers multiple opportunities to clarify the immunological/pathogenic role of CRP in health and disease.

7.
Can Urol Assoc J ; 9(5-6): E321-3, 2015.
Article in English | MEDLINE | ID: mdl-26029307

ABSTRACT

A 73-year-old man was admitted with progressive dyspnea; he also had benign prostatic hyperplasia (BPH). An angio computed tomography scan showed pulmonary embolism with thrombi in both main pulmonary arteries. By duplex ultrasonography, we detected a thrombus in the right vena femoralis superficialis and vena femoralis communis. Simultaneously, we also noticed a large diverticulum on the right side of the urinary bladder and urinary stasis II of the left kidney. We consider the BPH as the trigger for a secondary diverticulum of the urinary bladder. As a result of its large dimensions, mechanical compression of the deep right pelvic veins resulted in thrombosis which finally caused the pulmonary embolism. With respect to the urinary stasis II, surgical excavation of the diverticulum with infravesical desobstruction was planned. The potentially lethal course of large diverticula may require surgery.

8.
Mediators Inflamm ; 2014: 561428, 2014.
Article in English | MEDLINE | ID: mdl-24799767

ABSTRACT

The role of C-reactive protein (CRP) in atherosclerosis is controversially discussed. Whereas initial experimental studies suggested a pathogenic role for CRP in atherogenesis, more recent genetic data from Mendelian randomization trials failed to provide evidence for a causative role of CRP in cardiovascular disease. Also, experimental results from laboratories all over the world were indeed contradictory, partly because of species differences in CRP biology and partly because data were not accurately evaluated. Here we summarize the published data from experimental work with mainly human material in order to avoid confusion based on species differences in CRP biology. Experimental work needs to be reevaluated after reconsideration of some traditional rules in research: (1) in order to understand a molecule's role in disease it may be helpful to be aware of its role in physiology; (2) it is necessary to define the disease entity that experimental CRP research deals with; (3) the scientific consensus is as follows: do not try to prove your hypothesis. Specific CRP inhibition followed by use of CRP inhibitors in controlled clinical trials may be the only way to prove or disprove a causative role for CRP in cardiovascular disease.


Subject(s)
Atherosclerosis/metabolism , C-Reactive Protein/metabolism , Cardiovascular Diseases/metabolism , Humans
11.
J Am Heart Assoc ; 1(6): e003293, 2012 Dec.
Article in English | MEDLINE | ID: mdl-23316321

ABSTRACT

BACKGROUND: Overexpression of interleukin-10 (IL-10) in murine CD11b(+) monocytes/macrophages via GMP-adapted mRNA-nucleofection was expected to improve clinical outcome and reduce adverse side effects in autoimmune myocarditis. This study represents the proof of principle for a novel anti-inflammatory therapy using overexpression of IL-10 in murine monocytes/macrophages by mRNA-nucleofection for the treatment of autoimmune myocarditis. METHODS AND RESULTS: Autoimmune myocarditis was induced in A/J mice by subcutaneous immunization with troponin I. CD11b(+) monocytes/macrophages were isolated from the peritoneum and IL-10 was overexpressed by mRNA-nucleofection. These cells were injected intravenously. Myocardial inflammation was assessed via histological and immunohistochemical examinations. Myocardial fibrosis was analyzed with Masson's trichrome staining. Antitroponin I antibodies were determined within the serum. Physical performance was evaluated using a running wheel and echocardiography. In vitro overexpression of IL-10 in CD11b(+) monocytes/macrophages resulted in a 7-fold increased production of IL-10 (n=3). In vivo higher levels of IL-10 and less inflammation were detected within the myocardium of treated compared with control mice (n=4). IL-10-treated mice showed lower antitroponin I antibodies (n=10) and a better physical performance (n=10). CONCLUSIONS: Application of IL-10-overexpressing CD11b(+) monocytes/macrophages reduced inflammation and improved physical performance in a murine model of autoimmune myocarditis. Thus, the use of genetically modified monocytes/macrophages facilitated a targeted therapy of local inflammation and may reduce systemic side effects. Because the nucleofection technique is GMP adapted, an in vivo use in humans seems basically feasible and the transfer to other inflammatory diseases seems likely.


Subject(s)
Anti-Inflammatory Agents/therapeutic use , Autoimmune Diseases/therapy , Interleukin-10/therapeutic use , Myocarditis/therapy , Animals , Anti-Inflammatory Agents/blood , Anti-Inflammatory Agents/metabolism , Autoimmune Diseases/metabolism , Autoimmune Diseases/physiopathology , Cell Movement , Cells, Cultured , Enzyme-Linked Immunosorbent Assay , Feasibility Studies , Female , Gene Expression , Humans , Inflammation/metabolism , Inflammation/physiopathology , Inflammation/therapy , Interleukin-10/blood , Interleukin-10/genetics , Interleukin-10/metabolism , Macrophages/metabolism , Macrophages/physiology , Mice , Monocytes/metabolism , Monocytes/physiology , Myocarditis/metabolism , Myocarditis/physiopathology , RNA, Messenger , Transfection
12.
Clin Infect Dis ; 50(9): 1249-51, 2010 May 01.
Article in English | MEDLINE | ID: mdl-20367227

ABSTRACT

We report on 2 critically ill patients with pneumonitis and acute respiratory distress syndrome due to pandemic (H1N1) 2009 influenza A virus who were treated with intravenous zanamivir and had favorable clinical outcomes. Zanamivir given intravenously may be a therapeutic option in patients with critical illness and mechanical ventilation.


Subject(s)
Antiviral Agents/therapeutic use , Influenza A Virus, H1N1 Subtype/isolation & purification , Influenza, Human/complications , Influenza, Human/drug therapy , Pneumonia/drug therapy , Zanamivir/therapeutic use , Adult , Antiviral Agents/administration & dosage , Critical Illness , Humans , Influenza, Human/virology , Injections, Intravenous , Male , Middle Aged , Respiratory Distress Syndrome/drug therapy , Treatment Outcome , Zanamivir/administration & dosage
13.
Contrast Media Mol Imaging ; 5(2): 59-69, 2010.
Article in English | MEDLINE | ID: mdl-20235120

ABSTRACT

Various contrast agents (Magnevist, Gadovist and Multihance) loaded into polymeric nanocapsules were synthesized by the inverse miniemulsion technique. The relaxivity of the resultant contrast agents was assessed at 1.5 T magnetic field strength. The ionic relaxivity of the contrast agents could at least be maintained after their encapsulation in different polymer capsules. The chemical composition of the nanocapsules was characterized by Fourier transform infrared spectroscopy. The distribution of the contrast agent in the nanocapsules could be identified by energy filtered transmission electron microscopy and energy dispersive X-ray spectroscopy. The results indicate entrapment of the gadolinium complex into the inner shell of the polymeric nanocapsules. The payload of contrast agent per nanocapsule resulted in some 2.5 x 10(6) Gd(3+) complexes yielding a particle-based relaxivity of 10.75 x 10(6) mM(-1) s(-1). Maintained or even slightly increased ionic relaxivity of the different contrast agents after encapsulation in combination with high payloads and the possibility of functionalization of the capsules' surface facilitate the application of the nanocapsules as promising targeted contrast agents for MRI.


Subject(s)
Contrast Media/chemistry , Magnetic Resonance Imaging/methods , Nanocapsules , Chemistry Techniques, Analytical , Gadolinium , Materials Testing , Nanocapsules/chemistry , Polymers/chemistry
14.
J Card Fail ; 16(4): 348-56, 2010 Apr.
Article in English | MEDLINE | ID: mdl-20350703

ABSTRACT

BACKGROUND: Myocardial biopsy can be used for the detection of viral genome in dilated cardiomyopathy (DCM). Pilot studies have previously reported beneficial effects on clinical outcome and safety of an antiviral therapy using interferon beta-1b in chronic viral DCM. METHODS AND RESULTS: Myocardial biopsies were taken from patients with DCM. Using polymerase chain reaction and Southern Blot analysis, viral genome could be detected in 49% of patients. In 42 patients with viral infection, off-label use with interferon beta-1b was initiated. A further 68 patients formed the control group. The outcome was evaluated after follow-up with echocardiography, exercise electrocardiogram, and New York Heart Association class. A total of 81 men and 29 women with a median left ventricular ejection fraction of 34% were included. The follow-up period was 36 months. In 33 (79%) patients with interferon beta-1b treatment, minor adverse reactions occurred, but no major adverse events were reported. No significant benefit for interferon beta-1b treatment on clinical outcome could be detected during follow-up. CONCLUSIONS: Off-label use with interferon beta-1b in patients with viral DCM is feasible and safe under routine clinical practice. Concerning the herein evaluated clinical outcome parameters, promising results from pilot studies could not be confirmed. High prevalence of parvovirus B19 (92%) might influence the results.


Subject(s)
Cardiomyopathy, Dilated/drug therapy , Cardiomyopathy, Dilated/virology , Genome, Viral , Interferon-beta/physiology , Interferon-beta/therapeutic use , Virus Diseases/drug therapy , Virus Diseases/virology , Adult , Aged , Cardiomyopathy, Dilated/genetics , Chronic Disease , Female , Follow-Up Studies , Genome, Viral/genetics , Humans , Interferon beta-1b , Longitudinal Studies , Male , Middle Aged , Multicenter Studies as Topic/trends , Randomized Controlled Trials as Topic/trends , Retrospective Studies , Virus Diseases/genetics , Young Adult
15.
Eur Heart J ; 30(16): 2011-8, 2009 Aug.
Article in English | MEDLINE | ID: mdl-19633015

ABSTRACT

AIMS: Clinical parameters are weak predictors of outcome in patients with idiopathic dilated cardiomyopathy (IDC). We assessed the prognostic value of cardiac magnetic resonance (CMR) parameters in addition to conventional clinical and electrocardiographic characteristics. METHODS AND RESULTS: One hundred and forty-one IDC patients were studied. QRS and QTc intervals were measured in 12-lead surface electrocardiogram. Patients were followed for median 1339 days, including 483 patient-years. The primary endpoint-cardiac death or sudden death-occurred in 25 (18%) patients, including 16 patients with cardiac death, 3 patients with sudden cardiac death (SCD), and 6 patients with ICD shock. Late gadolinium enhancement (LGE) was detected in 36 patients (26%). Kaplan-Meier survival analysis displayed QRS >110 ms (P = 0.010), the presence of LGE (P = 0.037), and diabetes mellitus (P < 0.001) as significant parameters for a worse outcome. Multivariable analysis revealed cardiac index (P < 0.001), right ventricular end-diastolic volume index (RVEDVI) (P = 0.006) derived from CMR imaging, the presence of diabetes mellitus (P = 0.006), and QRS >110 ms (P = 0.045) as significant predictors for the primary endpoint. CONCLUSION: Cardiac index and RVEDVI derived from CMR imaging in addition to QRS duration >110 ms from conventional surface ECG and diabetes mellitus provide prognostic impact for cardiac death and SCD in patients with IDC.


Subject(s)
Cardiomyopathy, Dilated/mortality , Electrocardiography , Magnetic Resonance Angiography , Adult , Aged , Cardiomyopathy, Dilated/diagnosis , Cardiomyopathy, Dilated/physiopathology , Female , Humans , Kaplan-Meier Estimate , Male , Middle Aged , Prognosis , Risk Factors
16.
Basic Res Cardiol ; 104(5): 591-9, 2009 Sep.
Article in English | MEDLINE | ID: mdl-19343415

ABSTRACT

Whereas C-reactive protein (CRP) is acknowledged as a cardiovascular risk marker, there is ongoing discussion about its role as a risk factor. Previous studies focused on the effects of CRP on ischaemic heart failure and atherosclerosis. In this study we investigated distribution of CRP, the Terminal Complement Complex (C5b-9) and macrophages (CD68) in the myocardium of patients suffering from non-ischaemic heart failure and their implication on clinical parameters. Endomyocardial biopsies were taken from 66 patients suffering from dilated cardiomyopathy (DCM). Biopsies were analysed by immunohistochemical and immunofluorescent staining for CRP, C5b-9 and CD68. Viral DNA/RNA for adenovirus, enterovirus, parvovirus B19 and human herpes virus 6 was detected by PCR and Southern blot analysis. Myocardial biopsy findings were correlated with plasma level of hsCRP and NT-proBNP as well as echocardiography, exercise test and NYHA class. In 18 (27%) patients, a positive staining for CRP and in 57 (86%) patients a positive staining for C5b-9 was detected. All patients showed myocardial infiltration with macrophages with an average of 39 cells/mm(2). CRP, C5b-9 and CD68 co-localised within the myocardium. No correlation was observed for inflammatory proteins and plasma level of hsCRP, NT-proBNP and clinical parameters. CRP is frequently present in the myocardium of patients suffering from DCM and co-localises with C5b-9 and macrophages. CRP may contribute to myocardial damage in DCM via activation of the complement system and chemotaxis of macrophages.


Subject(s)
C-Reactive Protein/analysis , Cardiomyopathy, Dilated/immunology , Heart Failure/immunology , Myocarditis/immunology , Myocardium/immunology , Adenoviridae/genetics , Adult , Aged , Antigens, CD/analysis , Antigens, Differentiation, Myelomonocytic/analysis , Biopsy , Cardiomyopathy, Dilated/diagnostic imaging , Cardiomyopathy, Dilated/virology , Complement Membrane Attack Complex/analysis , DNA, Viral/isolation & purification , Enterovirus/genetics , Exercise Test , Female , Heart Failure/diagnostic imaging , Heart Failure/virology , Herpesvirus 6, Human/genetics , Humans , Macrophages/immunology , Male , Middle Aged , Myocarditis/diagnostic imaging , Myocarditis/virology , Myocardium/pathology , Natriuretic Peptide, Brain/analysis , Natriuretic Peptide, Brain/blood , Parvovirus B19, Human/genetics , Peptide Fragments/analysis , Peptide Fragments/blood , RNA, Viral/isolation & purification , Ultrasonography
17.
J Cell Mol Med ; 11(3): 521-30, 2007.
Article in English | MEDLINE | ID: mdl-17635643

ABSTRACT

Gene transfer into human CD34+ haematopoietic progenitor cells (HPC) and multi-potent mesenchymal stromal cells (MSC) is an essential tool for numerous in vitro and in vivo applications including therapeutic strategies, such as tissue engineering and gene therapy. Virus based methods may be efficient, but bear risks like tumorigenesis and activation of immune responses. A safer alternative is non-viral gene transfer, which is considered to be less efficient and accomplished with high cell toxicity. The truncated low affinity nerve growth factor receptor (ALNGFR) is a marker gene approved for human in vivo application. Human CD34+ HPC and human MSC were transfected with in vitro-transcribed mRNA for DeltaLNGFR using the method of nucleofection. Transfection efficiency and cell viability were compared to plasmid-based nucleofection. Protein expression was assessed using flow cytometry over a time period of 10 days. Nucleofection of CD34+ HPC and MSC with mRNA resulted in significantly higher transfection efficiencies compared to plasmid transfection. Cell differentiation assays were performed after selecting DeltaLNGFR positive cells using a fluorescent activating cell sorter. Neither cell differentiation of MSC into chondrocytes, adipocytes and osteoblasts, nor differentiation of HPC into burst forming unit erythroid (BFU-E) colony forming unit-granulocyte, erythrocyte, macrophage and megakaryocyte (CFU-GEMM), and CFU-granulocyte-macrophage (GM) was reduced. mRNA based nucleofection is a powerful, highly efficient and non-toxic approach for transient labelling of human progenitor cells or, via transfection of selective proteins, for transient manipulation of stem cell function. It may be useful to transiently manipulate stem cell characteristics and thus combine principles of gene therapy and tissue engineering.


Subject(s)
RNA, Messenger/genetics , RNA, Messenger/metabolism , Receptor, Nerve Growth Factor/biosynthesis , Receptor, Nerve Growth Factor/genetics , Stem Cells/metabolism , Transfection/methods , Antigens, CD34/metabolism , Cell Differentiation , Flow Cytometry , Humans , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/metabolism , Microscopy, Fluorescence , Plasmids , Stem Cells/cytology
18.
Eur J Heart Fail ; 9(1): 51-4, 2007 Jan.
Article in English | MEDLINE | ID: mdl-16730227

ABSTRACT

BACKGROUND: In patients with dilated cardiomyopathy (DCM), elevated plasma levels of tumor necrosis factor-alpha (TNF-alpha) are associated with poor prognosis. The terminal complement complex (C5b-9) stimulates myocardial TNF-alpha expression. AIMS: To investigate whether myocardial TNF-alpha and C5b-9 expression correlate with clinical outcome in DCM. METHODS AND RESULTS: 71 patients with DCM underwent myocardial biopsy. Biopsies were analyzed for TNF-alpha, C5b-9, markers of inflammation and for viral genome. Patients were divided into three groups according to biopsy results: group A: no TNF-alpha and no C5b-9; group B: TNF-alpha or C5b-9; and group C: TNF-alpha and C5b-9. NYHA classification, ECG and echocardiography were documented. Patients received conventional treatment of heart failure and, in a few cases, additional treatment with interferon beta(1b) (virus positive) or prednisolone (inflammatory DCM). There were 13 patients (18%) in group A, 19 patients (27%) in group B, and 39 patients (55%) in group C. All groups had a similar and significant improvement in NYHA classification and echocardiographic parameters. TNF-alpha and C5b-9 did not significantly correlate with the presence of viral genome or with markers of inflammation. CONCLUSION: TNF-alpha and C5b-9 are widely distributed in the myocardium of DCM patients. Neither of the antigens correlates with clinical outcome. Myocardial TNF-alpha may not be a useful prognostic marker in DCM.


Subject(s)
Cardiomyopathy, Dilated/blood , Complement Membrane Attack Complex/metabolism , Heart/physiopathology , Myocardium/metabolism , Tumor Necrosis Factor-alpha/metabolism , Adult , Aged , Biopsy , Cardiomyopathy, Dilated/pathology , Echocardiography , Electrocardiography , Female , Humans , Inflammation , Male , Middle Aged , Myocardium/pathology , Prognosis
19.
J Vasc Res ; 43(2): 157-65, 2006.
Article in English | MEDLINE | ID: mdl-16407661

ABSTRACT

BACKGROUND: Smooth muscle cell (SMC) proliferation in atherosclerosis is regulated through the interaction of growth factors like platelet-derived growth factor-BB (PDGF-BB) and insulin-like growth factor-1 (IGF-1) and their receptors (R). We hypothesized that serum starvation of SMCs may affect PDGFbeta-R and IGF-1-R expression and, consequently, the effect of their cognate ligands on SMC survival/proliferation. METHODS AND RESULTS: Serum starvation significantly increases PDGFbeta-R but not IGF-1-R mRNA and protein expression in SMCs. PDGF-BB stimulates cell survival but not proliferation in serum-starved SMCs of the synthetic phenotype, whereas SMCs of the contractile phenotype respond to PDGF-BB by a significant increase in proliferation. Immunohistochemical analysis of coronary atherosclerotic lesions reveals PDGFbeta-R expression in SMCs in the lamina fibromuscularis, but not in the media and in healthy parts of the arterial wall. No such differential expression was observed for IGF-1-R. CONCLUSIONS: Differential regulation of PDGFbeta-R and IGF-1-R expression by serum starvation might represent a mechanism for the control of SMC survival/proliferation in atherogenesis and restenosis. The distribution of PDGFbeta-Rs and IGF-1-Rs in atherosclerotic lesions may indicate an effect of serum starvation on SMCs in the arterial wall.


Subject(s)
Muscle, Smooth, Vascular/cytology , Muscle, Smooth, Vascular/physiopathology , Receptor, IGF Type 1/biosynthesis , Receptor, Platelet-Derived Growth Factor beta/biosynthesis , Becaplermin , Cell Proliferation , Cells, Cultured , Coronary Artery Disease/pathology , Coronary Artery Disease/physiopathology , Culture Media, Serum-Free , Down-Regulation , Humans , Platelet-Derived Growth Factor/pharmacology , Proto-Oncogene Proteins c-sis , Up-Regulation
20.
Eur J Heart Fail ; 8(2): 162-6, 2006 Mar.
Article in English | MEDLINE | ID: mdl-16111918

ABSTRACT

BACKGROUND: In some patients suffering from dilated cardiomyopathy (DCM) magnetic resonance imaging (MRI) shows late gadolinium enhancement with variable distribution. Myocardial biopsies in DCM reveal a chronic myocardial inflammatory process in almost 50% and myocardial persistence of adenoviral or enteroviral genome in about 15% of the patients. AIMS: We prospectively investigated whether the pattern of late gadolinium enhancement correlates with myocardial biopsy findings. METHODS AND RESULTS: 42 patients with DCM and 42 control subjects underwent contrast MRI. In the DCM group, endomyocardial biopsies were performed and evaluated for inflammation and viral genome. None of the control subjects showed late gadolinium enhancement whereas in 29 DCM patients (69%) gadolinium enhancement was detectable (p<0.001). 21 of the DCM patients (50%) showed midwall septal enhancement, 7 patients (17%) showed a patchy distribution of hyperenhancement and 1 patient (2%) showed enhancement typical for ischemic heart disease. In myocardial biopsy analysis, 2 patients (5%) showed persistence of viral genome, 18 patients (43%) showed inflammation and in 22 patients (52%) neither virus nor inflammation was detected. The pattern of late gadolinium enhancement and myocardial biopsy findings were not significantly correlated (p = 0.854). CONCLUSION: MRI as a non-invasive technique cannot replace myocardial biopsy for the differential diagnosis of DCM.


Subject(s)
Cardiomyopathy, Dilated/diagnosis , Cardiomyopathy, Dilated/physiopathology , Gadolinium , Myocardium/pathology , Adult , Aged , Biopsy , Cardiomyopathy, Dilated/pathology , Case-Control Studies , Female , Humans , Image Enhancement , Inflammation/physiopathology , Magnetic Resonance Imaging , Male , Middle Aged , Myocardium/ultrastructure , Prospective Studies
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