A physics-informed deep learning framework for dynamic susceptibility contrast perfusion MRI.

BACKGROUND: Perfusion magnetic resonance imaging (MRI)s plays a central role in the diagnosis and monitoring of neurovascular or neurooncological disease. However, conventional processing techniques are limited in their ability to capture relevant characteristics of the perfusion dynamics and suffer from a lack of standardization. PURPOSE: We propose a physics-informed deep learning framework which is capable of analyzing dynamic susceptibility contrast perfusion MRI data and recovering the dynamic tissue response with high accuracy. METHODS: The framework uses physics-informed neural networks (PINNs) to learn the voxel-wise TRF, which represents the dynamic response of the local vascular network to the contrast agent bolus. The network output is stabilized by total variation and elastic net regularization. Parameter maps of normalized cerebral blood flow (nCBF) and volume (nCBV) are then calculated from the predicted residue functions. The results are validated using extensive comparisons to values derived by conventional Tikhonov-regularized singular value decomposition (TiSVD), in silico simulations and an in vivo dataset of perfusion MRI exams of patients with high-grade gliomas. RESULTS: The simulation results demonstrate that PINN-derived residue functions show a high concordance with the true functions and that the calculated values of nCBF and nCBV converge towards the true values for higher contrast-to-noise ratios. In the in vivo dataset, we find high correlations between conventionally derived and PINN-predicted perfusion parameters (Pearson's rho for nCBF: 0.84 ± 0.03 $0.84 \pm 0.03$ and nCBV: 0.92 ± 0.03 $0.92 \pm 0.03$ ) and very high indices of image similarity (structural similarity index for nCBF: 0.91 ± 0.03 $0.91 \pm 0.03$ and for nCBV: 0.98 ± 0.00 $0.98 \pm 0.00$ ). CONCLUSIONS: PINNs can be used to analyze perfusion MRI data and stably recover the response functions of the local vasculature with high accuracy.

Introduction of inverse dose optimization for ultrasound-based high-dose-rate boost brachytherapy: how we do it in Kiel.

OBJECTIVES: To describe the introduction of inverse planning optimization for a two clinical target volume (CTV) concept in the online planning technique of temporary high-dose-rate brachytherapy for prostate cancer. METHODS AND MATERIALS: Dose-volume constraints were defined delivering a prescription dose of 8.5Gy for CTV1 (whole prostate) and 15Gy for CTV2 (peripheral zone). A total of 38 implants of 20 patients were inversely planned using the constraints and dose indices (D90 CTV1,2; V200 CTV1,2; D2 cc rectum; D0.1 cc urethra; dose nonhomogeneity ratio; and conformal index) compared against those derived from conventional planning (CP). RESULTS: The inversely planned (IP) treatment plans showed similar target volume coverage than by CP. The value of D90 CTV1 for CP was 5.62Gy and 5.63Gy for IPs. For CTV2, the D90 was also similar between both methods: 11.03Gy and 10.89Gy, respectively. Only V200 CTV2 was found to be significantly lower for CP than for IP: 5.76% vs. 8.14% (p<0.01). Values for D0.1 cc urethra were found to be: 9.57Gy and 9.02Gy, respectively. Rectal dosimetry: D2 cc Rectum was quite stable with 6.04Gy and 6.12Gy for CP and IP, respectively. The conformal index and dose nonhomogeneity ratio values for CTV1 and CTV2 for both planning types were very similar. CONCLUSIONS: After defining an objective second target volume CTV2 and introducing adequate IP constraints to the treatment planning system, clinically applicable treatment plans could be created by an IP approach. They feature user independency, time saving, and good preservation of the OARs.

Head and neck (192)Ir HDR-brachytherapy dosimetry using a grid-based Boltzmann solver.

PURPOSE: To compare dosimetry for head and neck cancer patients, calculated with TG-43 formalism and a commercially available grid-based Boltzmann solver. MATERIAL AND METHODS: This study included 3D-dosimetry of 49 consecutive brachytherapy head and neck cancer patients, computed by a grid-based Boltzmann solver that takes into account tissue inhomogeneities as well as TG-43 formalism. 3D-treatment planning was carried out by using computed tomography. RESULTS: Dosimetric indices D90 and V100 for target volume were about 3% lower (median value) for the grid-based Boltzmann solver relative to TG-43-based computation (p < 0.01). The V150 dose parameter showed 1.6% increase from grid-based Boltzmann solver to TG-43 (p < 0.01). CONCLUSIONS: Dose differences between results of a grid-based Boltzmann solver and TG-43 formalism for high-dose-rate head and neck brachytherapy patients to the target volume were found. Distinctions in D90 of CTV were low (2.63 Gy for grid-based Boltzmann solver vs. 2.71 Gy TG-43 in mean). In our clinical practice, prescription doses remain unchanged for high-dose-rate head and neck brachytherapy for the time being.

[79-year-old patient with pulmorenal syndrome]. / Pulmorenales Syndrom bei einer 79-jährigen Patientin.

HISTORY AND ADMISSION FINDINGS: A 79-year-old female patient was referred with acute renal failure requiring haemodialysis and haemoptysis. INVESTIGATIONS: Kidney biopsy showed extracapillary proliferative glomerulonephritis with crescents in 7 from 15 glomeruli and sclerosis in the remaining. A computed tomography scan of the chest showed evidence of alveolar haemorrhage. Serologic testing revealed autoantibodies against the glomerular basement membrane (anti-GBM antibodies) and myeloperoxidase antineutrophil cytoplasmic antibodies (MPO-ANCA). DIAGNOSIS, TREATMENT AND COURSE: The patient was diagnosed with goodpasture's disease and underwent immunosuppressive therapy including prednisolone, cyclophosphamide pulses and plasmapheresis, resulting in clearance of anti-GBM antibodies and discontinuation of haemoptysis. Renal function, however, did not recover and the patient remained on dialysis. CONCLUSIONS: Aggressive treatment including cyclophosphamide and plasma separation often ensures patient survival in goodpasture's disease, in most cases, however, renal function does not recover.

[Interpretation of elevated serum troponin levels in end stage renal disease - case 2/2010]. / Bedeutung erhöhter Troponin-Werte bei terminaler Niereninsuffizienz - Fall 2/2010.

HISTORY AND ADMISSION FINDINGS: We report on a female patient with rheumatoid arthritis and end-stage renal-disease following AA-amyloidosis who presented with chest pain to the emergency department. INVESTIGATIONS: ECG showed no signs of ischemia, echocardiography revealed a concentric left ventricular hypertrophy with increased texture. Serum concentration of troponin I was mildly elevated whereas creatine kinase (CK)/ CK-MB were normal. DIAGNOSIS, TREATMENT AND COURSE: The chief complaints resolved spontaneously and there was no change in the serum troponin-I and CK/CK-MB concentrations. Coronary heart disease was ruled out by angiography and cardiac involvement of the underlying AA-amyloidosis was diagnosed. After one month, the patient suffered from a syncope complicated by a pelvic ring fracture with hemorrhagic shock and declined chronic dialysis treatment. CONCLUSION: Patients with end-stage renal disease may exhibit a persisting elevation of serum troponin concentration reflecting the high burden of cardiovascular disease. Myocardial infarction can be distinguished by the lack of increase in serial tests.

Expression of blood coagulation factors on monocytes after exposure to TNF-treated endothelium in a novel whole blood model of arterial flow.

Activated blood monocytes are a major source of tissue factor (TF), the principal initiator of blood coagulation. TF can be shed from the monocyte surface in association with microparticles (MPs) and increased numbers of circulating MPs are indicative of poor clinical outcome in a number of inflammatory disorders, including atherosclerosis. The mechanisms coupling inflammation with aberrant TF production/activity remain obscure but the protease-activated receptor (PAR) family has been implicated. We have previously shown (i) that freshly isolated human monocytes express low levels of cell surface PAR-2, (ii) that cell surface PAR-2 is rapidly upregulated from intracellular stores following mechanical stimulation, and (iii) that PAR-2 stimulation results in elevation of intracellular calcium and cytokine release. Here, we have investigated the expression of PAR-2 on monocytes exposed to TNF-activated endothelial cells both under static conditions and in our newly-established model of arterial flow, using diluted whole blood. Monocyte surface PAR-2 expression was upregulated following static exposure to activated EC and with laminar (atheroprotective) arterial flow there was a further increase in monocyte PAR-2 expression. We have also shown under arterial flow conditions that exposure to TNF-stimulated EC resulted in a significant increase in expression of TF on monocytes compared to that on cells exposed to quiescent EC. These data strongly suggest that direct or indirect interactions with inflamed EC can modulate expression of PAR-2 and TF on the monocyte cell surface.

The epithelial membrane protein 1 is a novel tight junction protein of the blood-brain barrier.

In the central nervous system, a constant microenvironment required for neuronal cell activity is maintained by the blood-brain barrier (BBB). The BBB is formed by the brain microvascular endothelial cells (BMEC), which are sealed by tight junctions (TJ). To identify genes that are differentially expressed in BMEC compared with peripheral endothelial cells, we constructed a subtractive cDNA library from porcine BMEC (pBMEC) and aortic endothelial cells (AOEC). Screening the library for differentially expressed genes yielded 26 BMEC-specific transcripts, such as solute carrier family 35 member F2 (SLC35F2), ADP-ribosylation factor-like 5B (ARL5B), TSC22 domain family member 1 (TSC22D1), integral membrane protein 2A (ITM2A), and epithelial membrane protein 1 (EMP1). In this study, we show that EMP1 transcript is enriched in pBMEC compared with brain tissue and that EMP1 protein colocalizes with the TJ protein occludin in mouse BMEC by coimmunoprecipitation and in rat brain vessels by immunohistochemistry. Epithelial membrane protein 1 expression was transiently induced in laser-capture microdissected rat brain vessels after a 20-min global cerebral ischemia, in parallel with the loss of occludin immunoreactivity. The study identifies EMP1 as a novel TJ-associated protein of the BBB and suggests its potential role in the regulation of the BBB function in cerebral ischemia.

Expression of endothelial intercellular adhesion molecule-1 is determined by genotype: effects on efficiency of leukocyte adhesion to human endothelial cells.

Two biallelic polymorphisms, previously described in the human intercellular adhesion molecule (ICAM)-1 gene at codon 241 (glycine [G] to arginine [R] substitution) and codon 469 (glutamic acid [E] to lysine [K] substitution) have been associated with a number of diseases including myocardial infarction, transplant rejection, and diabetes. However, the functional significance of these polymorphisms has not been determined. ICAM-1 cell surface expression and ICAM-1-mediated leukocyte adhesion were investigated using Cos7 transfected with ICAM-1 polymorphic variants or human umbilical vein endothelial cells (HUVEC) of different ICAM-1 genotypes. There was significantly higher expression of surface ICAM-1 on Cos7 transfected with a plasmid encoding the GE (G241/E469) ICAM-1 variant or untreated HUVEC of GEGE (G241/E469 homozygous genotype). ICAM-1-mediated adhesion of peripheral blood mononuclear cells (PBMC) to GE-Cos7 cells or TNF-treated GEGE HUVEC was significantly increased. However, there was no significant difference in adhesion of PBMC to recombinant ICAM-1 of each polymorphic variant plated onto plastic wells. We conclude that the GE genotype of ICAM-1 is associated with greater cell surface expression of ICAM-1, which in turn leads to greater adhesion of leukocytes. This may explain the previously described associations of ICAM-1 polymorphisms with chronic inflammatory disease.

Proteome analysis of human substantia nigra in Parkinson's disease.

BACKGROUND: Parkinson's disease (PD) is the most common neurodegenerative disorder involving the motor system. Although not being the only region involved in PD, affection of the substantia nigra and its projections is responsible for some of the most debilitating features of the disease. To further advance a comprehensive understanding of nigral pathology, we conducted a tissue based comparative proteome study of healthy and diseased human substantia nigra. RESULTS: The gross number of differentially regulated proteins in PD was 221. In total, we identified 37 proteins, of which 16 were differentially expressed. Identified differential proteins comprised elements of iron metabolism (H-ferritin) and glutathione-related redox metabolism (GST M3, GST P1, GST O1), including novel redox proteins (SH3BGRL). Additionally, many glial or related proteins were found to be differentially regulated in PD (GFAP, GMFB, galectin-1, sorcin), as well as proteins belonging to metabolic pathways sparsely described in PD, such as adenosyl homocysteinase (methylation), aldehyde dehydrogenase 1 and cellular retinol-binding protein 1 (aldehyde metabolism). Further differentially regulated proteins included annexin V, beta-tubulin cofactor A, coactosin-like protein and V-type ATPase subunit 1. Proteins that were similarly expressed in healthy or diseased substantia nigra comprised housekeeping proteins such as COX5A, Rho GDI alpha, actin gamma 1, creatin-kinase B, lactate dehydrogenase B, disulfide isomerase ER-60, Rab GDI beta, methyl glyoxalase 1 (AGE metabolism) and glutamine synthetase. Interestingly, also DJ-1 and UCH-L1 were expressed similarly. Furthermore, proteins believed to serve as internal standards were found to be expressed in a constant manner, such as 14-3-3 epsilon and hCRMP-2, thus lending further validity to our results. CONCLUSION: Using an approach encompassing high sensitivity and high resolution, we show that alterations of SN in PD include many more proteins than previously thought. The results point towards a heterogeneous aetiopathogenesis of the disease, including alterations of GSH-related proteins as well as alterations of proteins involved in retinoid metabolism, and they indicate that proteins involved in familial PD may not be differentially regulated in idiopathic Parkinson's disease.

Major differences in gene expression in human coronary smooth muscle cells after nebivolol or metoprolol treatment.

OBJECTIVE: Vascular smooth muscle cells play a pivotal role in all stages of atherogenesis. Targeting their inflammatory and proliferative qualities might therefore inhibit the progression of atherosclerosis. This study aimed to characterize and compare the effects of the beta-receptor antagonists nebivolol and metoprolol on gene expression in human coronary artery smooth muscle cells (hcaSMC). METHODS AND RESULTS: hcaSMC were incubated with nebivolol or metoprolol (10(-5) mol/l) for 72 h. The downregulated genes are involved in inflammatory processes, oxidative stress and smooth muscle cell proliferation: i.e. downregulated were by nebivolol: interleukin-1alpha, cyclooxygenase-2, tumor-necrosis-factor (TNF)-alpha-induced protein 6, PDGF-A, growth-related oncogenes 2 and 3. Metoprolol increased the expression of interleukin-1alpha, cyclooxygenase-1, TNF-alpha-induced protein 3, heme oxygenase 1 and granulocyte/macrophage-colony-stimulating factor. In addition downregulated was monocyte chemoattractant protein 1 (MCP-1) mRNA by nebivolol. Nebivolol (10(-5) mol/l) reduced the amount of basal NF-kappaB after 48 and 52 h but not metoprolol. In the culture supernatants, MCP-1 concentrations were reduced by nebivolol. CONCLUSIONS: Nebivolol induced changes in the expression of inflammatory mediators in hcaSMC. These results add to data that suggest specific anti-inflammatory qualities of a beta-blocker of the third generation in comparison to metoprolol.

Influence of nebivolol and metoprolol on inflammatory mediators in human coronary endothelial or smooth muscle cells. Effects on neointima formation after balloon denudation in carotid arteries of rats treated with nebivolol.

OBJECTIVE AND BACKGROUND: Inflammation plays a critical role in all stages of atherogenesis. Proliferating vascular smooth muscle cells (SMC) and endothelial cells (EC) enhancing the inflammatory response, both contribute to the progression of atherosclerosis. Anti-proliferative, anti-inflammatory and anti-oxidative therapy seems to be a promising therapeutic strategy. The aim of this study was to assess the anti-proliferative and anti-inflammatory effect of the beta-blocker nebivolol in comparison to metoprolol in vitro and to find out whether nebivolol inhibits neointima formation in vivo. METHODS AND RESULTS: Real-time-RT-PCR revealed a decrease in VCAM-1, ICAM-1, PDGF-B, E-selectin and P-selectin mRNA expression in human coronary artery EC and SMC incubated with nebivolol for 72 hours while metoprolol did not have this effect. Nebivolol reduced MCP-1 and PDGF-BB protein in the culture supernatant of SMC and EC, respectively. Sprague-Dawley rats were treated with nebivolol for 0 or 35 days before and 28 days after carotid balloon injury. Immunohistological analyses showed that pre-treatment with nebivolol was associated with a decreased number of SMC layers and macrophages and an increased lumen area at the site of the arterial injury. The intima area was reduced by 43% after pre-treatment. CONCLUSION: We found that nebivolol reduced the expression of proinflammatory genes in endothelial cells and vascular smooth muscle cells in vitro whereas metoprolol did not. In vivo, nebivolol inhibited neointima formation by reducing SMC proliferation and macrophage accumulation.

Influence of growth factors on the proliferation of vascular smooth muscle cells isolated from subtotally nephrectomized rats after endothelin or angiotensin II antagonism.

BACKGROUND: Cardiovascular disease is the most important cause of death in patients with end-stage renal disease. In uraemia, the renin-angiotensin-aldosterone and endothelin (ET) systems are activated. It is not known whether inhibition of these systems attenuates the proliferation of isolated smooth muscle cells of uraemic rats. METHODS: Subtotally nephrectomized (SNX) rats were treated with an ET(A) receptor antagonist, an ET(AB) receptor antagonist, the angiotensin type 1 (AT1) receptor antagonist losartan (all 10 mg/kg body weight/day) or the angiotensin-converting enzyme (ACE) inhibitor trandolapril (0.1 mg/kg body weight/day) or received no medication (SNX) for 12 weeks. Then, aortal smooth muscle cells (SMCs) were isolated and cultivated. After incubation of SMCs with different growth factors (5-7 days), proliferation was measured using a bromodeoxyuridine enzyme-linked immunosorbent assay (BrdU ELISA). RESULTS: Higher maximum levels of proliferation were found in SMCs from untreated SNX rats than in SMCs from control animals [platelet-derived growth factor-BB (PDGF-BB) 486.60+/-8.27 vs 346.74+/-4.60%, basic fibroblast growth factor (bFGF) 176.68+/-6.50 vs 123.71+/-1.49%, tumour necrosis factor-alpha (TNF-alpha) 153.38+/-10.16 vs 122.27+/-1.41%]. Treatment with ET receptor antagonists or losartan attenuated growth factor-stimulated proliferation (PDGF-BB: ET(A) receptor antagonist, 135.71+/-1.08%; ET(AB) receptor antagonist, 122.72+/-0.58%; losartan: 103.69+/-1.83%, n = 8). SMCs from trandolapril-treated rats showed an increased response (PDGF-BB 663.48+/-7.00%, n = 8). CONCLUSIONS: Treatment of SNX rats with ET receptor antagonists or losartan reduced growth factor-induced SMC proliferation in vitro. However, further investigations with uraemic patients have to clarify whether angiotensin or ET receptor antagonists inhibit the development of atherosclerosis.

Influence of endothelin receptor antagonism on smooth muscle cell proliferation after chronic renal failure.

Uremic patients suffer from an accelerated development of atherosclerosis. In uremia the angiotensin-aldosterone-endothelin system is activated. It is not known, however, whether endothelin receptor antagonism inhibits atherosclerosis in uremia. An experimental model of mild renal insufficiency is subtotal nephrectomy in rats. The aim of this study was to assess the proliferative response of vascular smooth muscle cells from rats that have undergone subtotal nephrectomy and are treated with an endothelin-A or combined endothelin-A/endothelin-B receptor antagonist to the proatherogenic growth factors platelet-derived growth factor-BB and basic fibroblast growth factor. For 12 weeks 5/6-nephrectomized rats were treated with the endothelin-A receptor antagonist LU 302146 or the combined endothelin-A/endothelin-B receptor antagonist LU 302872 (10 mg/kg bodyweight)or received no medication (subtotal nephrectomy). Aortal smooth muscle cells were isolated and cultivated. After incubation with platelet-derived growth factor-BB (10(-13)-10(-9) mol/L for 5 days) or basic fibroblast growth factor (10(-14)-10(-10) mol/L for 7 days) proliferation was measured using bromodeoxyuridine enzyme-linked immunosorbent assay. Both growth factors increased proliferation in vascular smooth muscle cells from untreated subtotally nephrectomized rats (platelet-derived growth factor-BB10(-9) mol/L: 487%, basic fibroblast growth factor 10(-10) mol/L:175%). Treatment with the endothelin-A receptor antagonist resulted in a reduced proliferation (platelet-derived growth factor-BB: 137%, basic fibroblast growth factor: 109%). After treatment with the combined endothelin-A/endothelin-B receptor antagonist findings were similar (platelet-derived growth factor-BB: 123%,basic fibroblast growth factor: 110%). These data demonstrate that chronic endothelin-A and combined endothelin-A/endothelin-B receptor antagonism inhibits vascular smooth muscle cell proliferation in rats treated with subtotal nephrectomy. This indicates a regulatory influence of these drugs on gene transcription and supports the importance of early treatment to inhibit coronary artery disease in uremic patients.

Expression of solute carrier 7A4 (SLC7A4) in the plasma membrane is not sufficient to mediate amino acid transport activity.

Member 4 of human solute carrier family 7 (SLC7A4) exhibits significant sequence homology with the SLC7 subfamily of human cationic amino acid transporters (hCATs) [Sperandeo, Borsani, Incerti, Zollo, Rossi, Zuffardi, Castaldo, Taglialatela, Andria and Sebastio (1998) Genomics 49, 230-236]. It is therefore often referred to as hCAT-4 even though no convincing transport activity has been shown for this protein. We expressed SLC7A4 in Xenopus laevis oocytes, but could not detect any transport activity for cationic, neutral or anionic amino acids or for the polyamine putrescine. In addition, human glioblastoma cells stably overexpressing a fusion protein between SLC7A4 and the enhanced green fluorescent protein (EGFP) did not exhibit an increased transport activity for l-arginine. The lack of transport activity was not due to a lack of SLC7A4 protein expression in the plasma membrane, as in both cell types SLC7A4-EGFP exhibited a similar subcellular localization and level of protein expression as functional hCAT-EGFP proteins. The expression of SLC7A4 can be induced in NT2 teratocarcinoma cells by treatment with retinoic acid. However, also for this endogenously expressed SLC7A4, we could not detect any transport activity for l-arginine. Our data demonstrate that the expression of SLC7A4 in the plasma membrane is not sufficient to induce an amino acid transport activity in X. laevis oocytes or human cells. Therefore, SLC7A4 is either not an amino acid transporter or it needs additional (protein) factor(s) to be functional.

Modulation of the beta-adrenergic receptor system of vascular smooth muscle cells in vitro and in vivo by chronically elevated endothelin-1 levels.

Endothelin-1 (ET-1) levels are chronically elevated in several cardiovascular diseases and correlate with an increased mortality. However, in contrast to acute biological activities such as vasoconstriction, little is known about long-term effects of ET-1. In this study we determined the effects of ET-1 on the beta(2)-adrenergic receptor (AR) system. Incubation of smooth muscle cells with ET-1 for 72 hr led to increased beta(2)AR density as determined by radioligand binding. Experiments with inhibitors of protein and RNA synthesis as well as RT-PCR revealed that beta(2)AR upregulation required de novo synthesis. In addition, protein kinase C but neither NO nor prostaglandin metabolism were involved in this effect. The enhanced expression of beta(2)AR was associated with an increased expression of its stimulatory G-protein and the receptor's ability to stimulate adenylyl cyclase. To study chronic effects of ET-1 in vivo, rats were infused with ET-1 for 3 weeks. Similarly as in cultured cells, prolonged ET-1 exposure led to increased betaAR expression in vivo. As a consequence, beta(2)AR-induced vasodilatation was increased in aortic rings from ET-1-treated animals. Our results therefore suggest that chronically elevated ET-1 levels in vitro and in vivo induce counterregulatory mechanisms by increasing betaARs that attenuate the vasoconstrictive effects of ET-1.