Abdominal pain in PNH has never been investigated by in-vivo imaging studies. With MRI, we aimed to assess mesenteric vessels flow and small bowel wall perfusion to investigate the ischemic origin of abdominal pain.
Materials and Methods
Six PNH patients with (AP) and six without (NOP) abdominal pain underwent MRI. In a blinded fashion, mean flow (MF, quantity of blood moving through a vessel within a second, in mL·s-1) and stroke volume (SV, volume of blood pumped out at each heart contraction, in mL) of Superior Mesenteric Vein (SMV) and Artery (SMA), areas under the curve at 60 (AUC60) and 90 seconds (AUC90) and Ktrans were assessed by two operators.
Mean total perfusion and flow parameters were lower in AP than in NOP group. AUC60: 84.81 ± 11.75 vs. 131.73 ± 18.89 (P < 0.001); AUC90: 102.33 ± 14.16 vs. 152.58 ± 22.70 (P < 0.001); Ktrans: 0.0346 min-1 ± 0.0019 vs. 0.0521 ± 0.0015 (P = 0.093 duodenum, 0.009 jejunum/ileum). SMV: MF 4.67 ml/s ± 0.85 vs. 8.32 ± 2.14 (P = 0.002); SV 3.85 ml ± 0.76 vs. 6.55 ± 1.57 (P = 0.02). SMA: MF 6.95 ± 2.61 vs. 11.2 ± 2.32 (P = 0.07); SV 6.52 ± 2.19 vs. 8.78 ± 1.63 (P = 0.07). We found a significant correlation between MF and SV of SMV and AUC60 (MF:ρ = 0.88, P < 0.001; SV: ρ = 0.644, P = 0.024), AUC90 (MF: ρ = 0.874, P < 0.001; SV:ρ = 0.774, P = 0.003) and Ktrans (MF:ρ = 0.734, P = 0.007; SV:ρ = 0.581, P = 0.047).
Citation: De Cobelli F, Pezzetti G, Margari S, Esposito A, Giganti F, Agostini G, et al. (2015) New Insights in Abdominal Pain in Paroxysmal Nocturnal Hemoglobinuria (PNH): A MRI Study. PLoS ONE 10(4): e0122832. https://doi.org/10.1371/journal.pone.0122832
Academic Editor: Jiani Hu, Wayne State University, UNITED STATES
Received: October 27, 2014; Accepted: February 15, 2015; Published: April 21, 2015
Copyright: © 2015 De Cobelli et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Data Availability: All relevant data are within the paper.
Funding: Alexion Pharmaceuticals Inc. provided financial support in the form of a grant. This does not alter the authors' adherence to PLOS ONE policies on sharing data and materials. Alexion Pharmaceuticals Inc. had no involvement in the study design, the collection and the analysis of the data and results.
Competing interests: The authors have declared that no competing interests exist.
Paroxysmal Nocturnal Hemoglobinuria (PNH) is an acquired clonal disorder of hematopoietic stem cells due to somatic mutations in the PIG-A gene, with an early block in the synthesis of the glycosylphosphatidylinositol (GPI) anchor on the cell membrane .
There is a deficiency of GPI-anchored proteins CD55 and CD59, inhibiting factors of the complement: red blood cells are prone to complement-induced lysis, leading to persistent intravascular hemolysis with brisk exacerbations [1,2].
In two retrospective studies of 220 and 460 PNH patients [6,7], the cumulative incidence rate of thromboembolic events was 30.7% with an incidence at the time of diagnosis of 7.2%  and a 10.2% relative risk of thrombosis .
The analysis of 195 PNH patients of three independent clinical studies and an open-label study [8–11] showed that 18.5% of the thromboembolic events regarded the mesenteric/splenic veins and 16.9% the portal vein [3,12]; thrombotic complications were the cause of death in the 44% of the patients .
Abdominal pain is one of the main causes of discomfort and disability in PNH and is present in approximately one-third of the patients at diagnosis ; it is associated with higher risk of thromboembolic events .
Albeit Magnetic Resonance Imaging (MRI) is considered the method of choice to assess vascular patency and parenchymal iron overload in PNH , the exact pathophysiology (i.e. the ischemic origin) has never been accurately investigated by in-vivo imaging studies.
Moreover, DCE-MRI parameters, area under the curve (AUC) and Ktrans, give important semi-quantitative/quantitative information about bowel wall perfusion. AUC is the area under the signal intensity curve from the time of contrast agent injection to usually 60 and 90 s .
Ktrans represents the product of tissue blood flow and the incomplete first-pass extraction fraction of the contrast agent from the vascular system into the extravascular/extracellular space.
The extraction fraction is a function of the capillary permeability and blood flow .
To date, DCE-MRI functional assessment of small bowel microvascular perfusion and mesenteric flow analysis have never been performed in PNH.
Thus, in order to investigate the ischemic origin of abdominal pain, our purpose was to assess both the mesenteric vessels flow and small bowel wall perfusion using MRI.
Materials and Methods
This is a prospective study, approved by the San Raffaele Hospital Scientific Institute Ethics Board.
All patients provided written informed consent.
Between April 2012 and July 2013, twelve patients with PNH (7 women and 5 men, aged from 21 to 59 years), untreated or previously treated with Eculizumab (suspended more than three months before enrolment) were enrolled based on their clinical history of presence (AP) or absence (NOP) of abdominal pain.
Eculizumab is a humanized monoclonal antibody preventing the assembly of the membrane attack complex of the complement.
Exclusion criteria were:
- MRI contraindication;
- Renal failure (Glomerular filtration rate < 30ml·min-1);
- Intolerance to contrast medium or Scopolamine-butylbromide;
- Therapy with Eculizumab suspended less than three months before enrolment;
- History of drug/alcohol abuse;
- Any condition/ongoing medication able to induce abdominal pain.
The diagnosis of PNH was based on the granulocyte PNH clone in the peripheral blood assessed by flow cytometry analysis (at least 20% [22–24]) and on the increase of serum Lactate Dehydrogenase (LDH > 1.5 ULN).
All NOP patients enrolled in this study did not experience any abdominal pain attributable to PNH.
In the AP group, patients had more than 4 episodes of abdominal pain per year, with at least one episode in the quarter preceding the enrolment.
None of the AP patients had pain at the time of MRI scan.
The association of the pain with food assumption, the average duration of the single episodes of pain, the type of presentation, the association with hemolytic attacks (LDH levels), the average intensity (from 0 to 10)  and the concurrent antalgic, anti-spastic and anti-coagulant/anti-platelet therapies were assessed.
All imaging studies were performed on a 1.5T MRI scanner (Achieva Nova; Philips Medical Systems, Best, the Netherlands) with high-performance gradients (maximum strength of 33 m·T·m1; slew rate of 150–180 m·T·m-1·s-1) and a 16-elements SENSE phased-array coil.
To ensure the homogenization of the intestinal activity and adequate luminal distension, all the subjects fasted for 6 hours and 1.5 L of a polyethylene glycol (PEG) solution was orally administered 30 minutes before the examination .
MRI protocol is summarized in Table 1.
With patients in the supine position, breath-hold (SSFP) sequences were acquired to visualize the superior mesenteric vein (SMV) and artery (SMA) and the small bowel wall and to exclude any other intestinal disease.
Flow mapping was performed with a breath-hold Electrocardiography (ECG)-gated Q-Flow 2D-PC FFE sequences acquired perpendicularly to SMV and SMA [26,27], using three different levels of maximum velocity encoding (VENC): the sequence with the lowest velocity and without aliasing phenomena was considered. Bipolar velocity encoding gradients were applied along the flow direction.
To minimize bowel peristalsis, 20 mg of Scopolamine-butylbromide were administered intravenously before perfusion data acquisition (in the absence of contraindications).
MR perfusion study was performed with a breath-hold 3D T1 GE sequence in the coronal plane, after intravenous administration of 0.1mmol/kg of paramagnetic contrast material (Gadobutrol) with an automatic injector (Spectris MR, Medrad Europe, Maastricht, The Netherlands) at a rate of 2 mL/s.
MRI Flow Analysis
Magnitude and phase MR images of the superior mesenteric artery and vein were displayed on an image-processing workstation (Extended Viewforum, Philips Medical System, Best, The Netherlands) with flow analysis package (MR Workspace 18.104.22.168). The luminal area was traced manually on the magnitude images, automatically transferred to the velocity phase images and adjusted according to the cardiac phase (Fig 1).
In Phase-contrast sequences, elliptic ROIs were manually positioned on the superior mesenteric vein (SMV) and artery (SMA) in order to obtain curves of velocity and flow rate versus time.
At various values of VENC (40, 80 and 120 cm·s-1), Mean Flow (MF, i.e. the quantity of blood moving through a vessel within a second, in mL·s-1) and Stroke Volume (SV, i.e. the volume of blood pumped out at each contraction of the heart, in mL) within each Region Of Interest (ROI) and the area of ROI (cm2) were determined for each cardiac frame and curves of velocity and flow rate versus time were automatically reconstructed.
MRI Perfusion analysis
Post-processing was performed with NordicICE Software 2.3.12 (Nordic Imaging Lab AS, Bergen, Norway).
Elliptic ROIs (diameters of 3 x 2 mm) were manually drawn in the wall of descending duodenum, proximal and distal jejunum and proximal, middle and distal ileum (Fig 2).
To calculate MRI perfusion parameters, AUC60, AUC90 and Ktrans, elliptic ROIs were positioned in different segments of the small bowel wall, particularly on proximal and distal jejunum and proximal, middle and distal ileum, as showed in this figure.
To ensure the homogeneity of the data, all ROIs were placed in the segments of the small bowel mentioned above and several values for each segment were collected before finally considering their mean value.
Qualitative, semi-quantitative and quantitative maps of the parameters related to vascular permeability and intra/extra vascular volumes (based on the dynamic effect of the contrast agent) were obtained.
Areas under the signal-intensity curve from the time of injection of Gadolinium to 60 and 90 s post-injection (AUC60 and AUC90) were calculated by cubic interpolation and digital integration.
All the measurements of flow and perfusion parameters were performed in blind by two operators (GP and FDC, respectively with 5 and 20 years of experience in abdominal MRI) who placed the ROIs and assessed measurements, blinded to all clinical data.
Data analysis was performed by using IBM SPSS Statistics software (version 20.0; SPSS, Chicago, Ill., USA).
All parameters were checked graphically for central tendency, spread and skew.
Due to the small and independent samples of patients, data were compared with Mann-Whitney Test for independent samples.
The relationships between flow and perfusion parameters were assessed with Spearman rank correlation test.
Inter-observer agreement was evaluated with Spearman rank correlation test.
A P value of less than 0.05 (two-tailed testing) indicated a statistically significant difference.
The review of the manuscript was performed by F.D.C., with a 20-year post-fellowship experience in abdominal MRI.
Clinical and biochemical parameters (i.e. age, sex, body mass index, arterial blood pressure, heart rate, years from PNH diagnosis, granulocytes and red cells PNH clone percentages, concentration of Hemoglobin, serum LDH levels, need of transfusions, pain score) are listed in Table 2.
All blood tests were performed at the time of MRI scan.
No significant differences were seen between the AP and NOP groups in terms of age (P = 0.59), BMI (P = 0.39), PNH clone in the granulocytes (P = 0.82), LDH serum levels (P = 0.59) and Hemoglobin (P = 0.82) (Table 2).
The mean DCE-MRI data both in each intestinal segment and in the whole small bowel were significantly lower in the AP than in the NOP group (Table 3).
In the whole small bowel AUC60 was 84.81 ± 4.99 in AP vs 131.73 ± 18.47 (P < 0.001) in NOP and AUC90 was 102.33 ± 5.76 vs 152.58 ± 23.11 (P < 0.001) (Fig 3).
The horizontal axis represents the patients without (0) and with (1) abdominal pain; the vertical axis represents the AUC60 (darker colors) and AUC90 (lighter colors) values in duodenum (blue), jejunum (green) and ileum (cyan). **: P<.01; Errors bars: 95% of confidence interval.
Ktrans was 0.0346 ± 0.0019 vs 0.0521 ± 0.0015 min-1 (P < 0.001) (Fig 4).
The horizontal axis represents the PNH patients without (0) and with (1) abdominal pain; the vertical axis represents the Ktrans values (min-1) in duodenum, jejunum and ileum. **: P<.01; Errors bars: 95% of confidence interval.
In the duodenum AUC60 was 81.26 ± 4.61 in AP vs 115.34 ± 11.34 in NOP (P = 0.002); AUC90 was 96.02 ± 11.42 vs 127.86 ± 13.55 (P = 0.002) (Fig 3); Ktrans was 0.0363 ± 0.0129 vs 0.0511 ± 0.0094 (P = 0.09) (Fig 4).
In the jejunum AUC60 was 90.51 ± 12.10 in AP vs 151.74 ± 12.52 in NOP (P = 0.002) and AUC90 was 107.29 ± 15.13 vs 173.65 ± 14.32 (P = 0.002) (Fig 3); Ktrans was 0.0350 ± 0.0067 vs 0.0538 ± 0.0084 (P = 0.009) (Fig 4).
In the ileum AUC60 was 82.66 ± 15.58 in AP vs 128.12 ± 10.46 in NOP (P = 0.002) and AUC90 was 103.69 ± 15.62 vs 156.24 ± 9.02 (P = 0.002) (Fig 3); Ktrans was 0.0325 ± 0.0034 vs 0.0514 ± 0.0123 (P = 0.009) (Fig 4).
Similarly, the mean blood flow MRI data (MF and SV) in the superior mesenteric artery (SMA) and vein (SMV) showed a lower flow in the AP than in the NOP group, respectively, even if a significant difference was found only in the venous compartment.
In SMV, MF was 4.67 ± 0.85 vs 8.32 ± 2.14 mL ·s-1 (P = 0.002) and SV 3.85 ± 0.76 vs 6.55 ± 1.57 mL (P = 0.02) for AP and NOP, respectively (Fig 5).
The horizontal axis represents the PNH patients without (0) and with (1) abdominal pain; the vertical axis represents the MF and SV values on superior mesenteric vein (SMV, dark blue) and artery (SMA, light blue). *: P<.05; **: P<.01; Errors bars: 95% of confidence interval.
In SMA, MF was 6.95 ± 2.61 vs 11.2 ± 2.32 (P = 0.07) and SV was 6.52 ± 2.19 vs 8.78 ± 1.63 (P = 0.07) for AP and NOP, respectively (Fig 5).
Inter-observer agreement was good both for perfusion and flow parameters in patients with pain (MF SMV:ρ = 0.943; P < 0.001; SV SMV:ρ = 0.986; P < 0.001; MF SMA:ρ = 0.943; P = 0.005; SV SMA: ρ = 0.986; P < 0.001; total AUC60: ρ = 0.957; P < 0.001; total AUC90: ρ = 0.944; P< 0.001; total Ktrans:ρ = 0.998; P < 0.001) and in patients without pain (MF SMV: ρ = 0.986; P < 0.001; SV SMV: ρ = 0.943; P < 0.001; MF SMA: ρ = 0.943; P < 0.001; SV SMA: ρ = 0.986; P < 0.001; total AUC60: ρ = 0.973; P < 0.001; total AUC90: ρ = 0.983; P < 0.001; total Ktrans: ρ = 0.991; P < 0.001).
Regarding the correlation analysis, we obtained these results:
- MF in SMV vs total AUC60:ρ = 0.88, P < 0.001 (Fig 6A); total AUC90:ρ = 0.874, P < 0.001 (Fig 6B); total Ktrans:ρ = 0.734, P = 0.007 (Fig 6C).
- SV of SMV vs total AUC60:ρ = 0.644, P = 0.024 (Fig 7A); total AUC90:ρ = 0.774, P = 0.003 (Fig 7B); total Ktrans:ρ = 0.581, P = 0.047 (Fig 7C).
- MF of SMA vs total AUC60:ρ = 0.546, P = 0.066 (Fig 8A); total AUC90:ρ = 0.459, P = 0.134 (Fig 8B); total Ktrans:ρ = 0.553, P = 0.062 (Fig 8C).
- SV of SMA vs total AUC60:ρ = 0.580, P = 0.048 (Fig 9A); total AUC90:ρ = 0.608, P = 0.036 (Fig 9B); total Ktrans:ρ = 0.580, P = 0.048 (Fig 9C).
Correlation between MRI perfusion parameters (on the horizontal axis)—AUC60 (A), AUC90 (B), and Ktrans (C)—in the whole small bowel—of PNH patients and MF of SMV (on the vertical axis).
Correlation between MRI perfusion parameters (on the horizontal axis)—AUC60 (A), AUC90 (B), and Ktrans (C)—in the whole small bowel—of PNH patients and SV of SMV (on the vertical axis).
Correlation between MRI perfusion parameters (on the horizontal axis)—AUC60 (A), AUC90 (B), and Ktrans (C)—in the whole small bowel—of PNH patients and MF of SMA (on the vertical axis).
Our prospective study suggests that both small bowel blood flow and perfusion impairment might be reliable MRI markers of mesenteric ischemia in untreated PNH patients with abdominal pain.
The thrombophilic tendency in PNH patients is a multifactorial phenomenon related to a consensual activation of platelets and complement system leading to endothelial dysfunction with thrombin generation and fibrinolytic defect .
Unchecked complement activity has a direct effect on platelets and can initiate thrombosis, which activates the complement system triggering a vicious thrombophilic cycle until the patient develops potentially lethal major thrombotic complications .
Of note, while intestinal ischemia has been postulated to be the cause of recurrent bouts of abdominal pain in patients with PNH [28,29], there are few data concerning a direct ante-mortem evidence of this aspect .
Dolezel et al. have interestingly investigated the presence of small bowel wall thickening (suggesting recurrent ischemia) in a single case of PNH with recurrent abdominal pain using Computed Tomography (CT) and MRI .
Furthermore, several studies [17,19,26,27] have previously evaluated the presence of mesenteric ischemia analyzing flow and perfusion by MRI, confirming a reduction of small-bowel perfusion and mesenteric venous flow.
Our study adds to the current literature by providing initial evidence of the differences in mesenteric flow and in small bowel wall perfusion in PNH patients with and without abdominal pain.
As a consequence, MR flow quantification on these vessels reflects the whole small intestine blood supply . Moreover, performing DCE-MRI (i.e. measuring the contrast-induced changes in tissue T1 relaxivity) allows to investigate microvessels density and capillary endothelial permeability [32,33].
In our study, we focused both on DCE semi-quantitative AUC (that represents the integrated area under the contrast medium concentration–time curve at different time points post contrast agent injection, in our case 60 and 90 s) and on the quantitative Ktrans, that reflects the two-compartment pharmacokinetic model of the contrast medium (intravascular and extra vascular components) .
Ktrans is dependent on flow and permeability-surface area.
Of note, we found that in PNH patients with abdominal pain, all flow parameters were significantly lower in the venous compartment together with the AUC and Ktrans values (especially in jejunum and ileum). In addition to this, low levels of MRI perfusion parameters in the whole small bowel were independently and strictly associated with low MF (P < 0.001) and SV (P < 0.05) values in SMV, confirming that in patients with PNH blood flow is strongly reduced in the venous district.
Conversely, we observed that the arterial compartment was less involved, as only SV of SMA, but not MF, was significantly associated with perfusion parameters (P < 0.05).
In order to investigate this interesting finding, it is realistic to assume that a microvascular damage occurs in PNH. In fact, the increased vascular resistance/density and the intrinsic low venous flow, might increase the time of contact/transit of pro-inflammatory mediators and reactive oxygen species along the intimal layer of the vessels.
As a result, this condition may lead to endothelial dysfunction, self-perpetuating the damage, as highlighted by some previous electron microscopy findings of capillaries with coarsely granular plasma and fibrin plugs occluding the lumen  and confirmed on MRI by our study (i.e. the low levels of perfusion parameters, especially Ktrans).
Moreover, nitric oxide scavenging/depletion and the burden of oxidative stress in the local vascular bed might cause constriction and spasm of the small peripheral mesenteric arterial vessels with transient ischemia and consequent reduction in the venous drainage of the mesenteric compartment.
This might lead to vascular dysfunction and microthrombosis, often associated with brisk crisis of abdominal pain, triggered by conditions that induce complement activation and the subsequent intravascular hemolysis.
We acknowledge an important limitation of our study, specifically the small number of patients: this is mainly due to the rarity of this pathology and the difficulty to recruit untreated PNH patients without any other concomitant disease.
Nevertheless, we deem that our results provide initial evidence of the importance of MRI as a reliable tool to analyze mesenteric ischemia in untreated PNH patients with abdominal pain and point out the need of larger prospective studies investigating the main purpose of this report.
Small bowel blood flow and perfusion impairment seem to be early and reliable MRI markers of mesenteric ischemia in untreated PNH patients with abdominal pain.
From a clinical point of view, our initial findings could be useful to select those patients who could benefit from an early tailored therapy with Eculizumab and to monitor treatment, as assessed through flow and perfusion changes before and after therapy.
We thank Prof. Andrea Falini for the kind permission to use NordicICE Software 2.3.12.
We are indebted to the following colleagues for the recruitment of the patients:
- Dr. Carlo Finelli, Department of Hematology, Oncology and Laboratory Medicine. Institute of Hematology Lorenzo and Ariosto Seragnoli. S. Orsola—Malpighi University Hospital of Bologna. Bologna, Italy.
- Dr. Francesco Caracciolo, Department of Hematology. Pisa University Hospital. Pisa, Italy.
- Dr. Anna Paola Iori, Department of Hematology—Umberto I Hospital, “La Sapienza” University. Rome, Italy.
- Dr. Massimiliano Bonifacio, Department of Hematology—Borgo Roma Hospital. Verona, Italy.
- Dr. Margherita Maffioli, Macchi Foundation Hospital. Varese, Italy.
- Dr. Francesco Lanza, Cremona Hospitals. Cremona, Italy.
- Dr. Wilma Barcellini, Ca 'Granda Foundation University Hospital. Milan, Italy.
- Dr. Fabio Montanelli, A. Manzoni Hospital. Lecco, Italy.
- Dr. Matteo Della Porta, San Matteo University Hospital. Pavia, Italy.
- Dr. Pietro Pioltelli, San Gerardo Hospital. Monza, Italy.
Conceived and designed the experiments: FDC. Performed the experiments: FDC GP SM AE GA. Analyzed the data: FDC GP SM AE GA. Wrote the paper: FDC GP SM AE GA. Checked the language revision: FG GA. Revised the manuscript: FG ADM.
- 1. Hernández-Campo PM, Almeida J, Sánchez ML, Malvezzi M, Orfao A. Normal patterns of expression of glycosylphosphatidylinositol-anchored proteins on different subsets of peripheral blood cells: a frame of reference for the diagnosis of paroxysmal nocturnal hemoglobinuria. Cytometry B Clin Cytom. 2006;70: 71–81. pmid:16493662
- 2. Holers VM. The spectrum of complement alternative pathway-mediated diseases. Immunol Rev. 2008;223: 300–316. pmid:18613844
- 3. Hill A, Kelly RJ, Hillmen P. Thrombosis in paroxysmal nocturnal hemoglobinuria. Blood. 2013;121: 4985–4996. pmid:23610373
- 4. Wang H, Chuhjo T, Yasue S, Omine M, Nakao S. Clinical significance of a minor population of paroxysmal nocturnal hemoglobinuria-type cells in bone marrow failure syndrome. Blood. 2002;100: 3897–3902. pmid:12393738
- 5. Gralnick HR, Vail M, McKeown LP, Merryman P, Wilson O, Chu I, et al. Activated platelets in paroxysmal nocturnal hemoglobinuria. Br J Haematol. 1995;91: 697–702. pmid:8555078
- 6. Socié G, Mary JY, de Gramont A, Rio B, Leporrier M, Rose C, et al. Paroxysmal nocturnal haemoglobinuria: long term follow-up and prognostic factors. Lancet. 1996;348: 573–577. pmid:8774569
- 7. Peffault de Latour R, Mary JY, Salanoubat C, Terriou L, Etienne G, Mohty M, et al. Paroxysmal nocturnal hemoglobinuria: natural history of disease subcategories. Blood. 2008;112: 3099–3106. pmid:18535202
- 8. Hillmen P, Muus P, Dursen U, Risitano AM, Schubert J, Luzzatto L, et al. Effect of complement inhibitor Eculizumab on thromboembolism in patients with paroxysmal nocturnal hemoglobinuria. Blood. 2007;110: 4123–4128. pmid:17702897
- 9. Hillmen P, Hall C, Marsh JC, Elebute M, Bombara MP, Petro BE, et al. Effect of eculizumab on hemolysis and transfusion requirements in patients with paroxysmal nocturnal hemoglobinuria. N Engl J Med. 2004;350: 552–559. pmid:14762182
- 10. Hillmen P, Young NS, Schubert J, Brodsky RA, Socié G, Muus P, et al. The complement inhibitor Eculizumab in paroxysmal nocturnal hemoglobinuria. N Engl J Med. 2006;355: 1233–1243. pmid:16990386
- 11. Young NS, Antonioli E, Rotoli B, Schrezenmeier H, Schubert J, Urbano-Ispizua A, et al. Safety and efficacy of the terminal complement inhibitor eculizumab in patients with paroxysmal nocturnal hemoglobinuria: Interim Shepherd Phase III Clinical Study [abstract]. Blood. 2006;108: 971.
- 12. Audebert HJ, Planck J, Eisenburg M, Schrezenmeier H, Haberl R. Cerebral ischemic infarction in paroxysmal nocturnal hemoglobinuria report of 2 cases and updated review of 7 previously published patients. J Neurol. 2005;252: 1379–1386. pmid:16021362
- 13. Crosby W. Paroxysmal nocturnal hemoglobinuria: a classic description by Paul Strubing in 1882 and a bibliography of the disease. Blood. 1951;6: 270. pmid:14811916
- 14. Lee JW, Jang JH, Kim JS, Yoon SS, Lee JH, Kim YK, et al. Clinical signs and symptoms associated with increased risk for thrombosis in patients with paroxysmal nocturnal hemoglobinuria from a Korean Registry. Int J Hematol. 2013;97: 749–757. pmid:23636668
- 15. Mathieu D, Rahmouni A, Villeneuve P, Anglade MC, Rochant H, Vasile N. Impact of Magnetic Resonance Imaging on the Diagnosis of Abdominal Complications of Paroxysmal Nocturnal Hemoglobinuria. Blood. 1995;85: 3283–3288. pmid:7756661
- 16. Grand DJ, Beland M, Harris A. Magnetic resonance enterography. Radiol Clin North Am. 2013;51: 99–112. pmid:23182510
- 17. Lauenstein TC, Ajaj W, Narin B, Göhde SC, Kröger K, Debatin JF, et al. MR imaging of apparent small-bowel perfusion for diagnosing mesenteric ischemia: feasibility study. Radiology. 2005;234: 569–575. pmid:15601890
- 18. Burkart DJ, Johnson CD, Reading CC, Ehman RL. MR measurements of mesenteric venous flow: prospective evaluation in healthy volunteers and patients with suspected chronic mesenteric ischemia. Radiology. 1995;194: 801–806. pmid:7862982
- 19. Naganawa S, Cooper TG, Jenner G, Potchen EJ, Ishigaki T. Flow velocity and volume measurement of superior and inferior mesenteric artery with cine phase-contrast magnetic resonance imaging. Radiat Med. 1994;12: 213–220. pmid:7863025
- 20. Giusti S, Faggioni L, Neri E, Fruzzetti E, Nardini L, Marchi S, et al. Dynamic MRI of small bowel: usefulness of quantitative contrast enhancement parameters and time-signal intensity curves for differentiating between active and inactive Crohn’s disease. Abdom Imaging. 2010;35: 646–653. pmid:20509025
- 21. Barnes SL, Whisenant JG, Loveless ME, Yankeelov TE. Practical dynamic contrast enhanced MRI in small animal models of cancer: data acquisition, data analysis, and interpretation. Pharmaceutics. 2012;4: 442–478. pmid:23105959
- 22. Borowitz MJ, Craig FE, Digiuseppe JA, Illingworth AJ, Rosse W, Sutherland DR, et al. Guidelines for the diagnosis and monitoring of paroxysmal nocturnal hemoglobinuria and related disorders by flow cytometry. Cytometry B, Clin Cytom. 2010;78: 211–230. pmid:20533382
- 23. Parker C, Omine M, Richards S, Nishimura J, Bessler M, Ware R, et al. Diagnosis and management of paroxysmal nocturnal hemoglobinuria. Blood. 2005;106: 3699–3709. pmid:16051736
- 24. Richards SJ, Hill A, Hillmen P. Recent Advances in the Diagnosis, Monitoring and Management of patients with Paroxysmal Nocturnal Haemoglobinuria. Cytometry B, Clin Cytom. 2007;72: 291–298. pmid:17549742
- 25. Cleeland CS, Ryan KM. Pain assessment: global use of the Brief Pain Inventory. Ann Acad Med Singapore. 1994;23: 129–138. pmid:8080219
- 26. Li KC, Hopkins KL, Dalman RL, Song CK. Simultaneous measurement of flow in the superior mesenteric vein and artery with cine phase-contrast MR imaging: value in diagnosis of chronic mesenteric ischemia. Radiology. 1995;194: 327–330. pmid:7824706
- 27. Li KC, Whitney WS, McDonnell CH, Fredrickson JO, Pelc NJ, Dalman RL, et al. Chronic mesenteric ischemia: evaluation with phase-contrast cine MR imaging. Radiology. 1994;190: 175–179. pmid:8259400
- 28. Blum SF, Gardner FH. Intestinal infarction in paroxysmal nocturnal hemoglobinuria. N Engl J Med. 1966;274: 1137.
- 29. Doukas M, Di Lorenzo P, Mohler D. Intestinal infarction caused by paroxysmal nocturnal hemoglobinuria. Am J Hematol. 1984; 16:75–81. pmid:6695911
- 30. Adams T, Fleischer D, Marino G, Rusnock E, Li L. Gastrointestinal involvement in paroxysmal nocturnal hemoglobinuria: first report of electron microscopic findings. Dig Dis Sci. 2002;47: 58–64. pmid:11837733
- 31. Dolezel Z, Dostalkova D, Blatny J, Starha J, Gerykova H. Paroxysmal nocturnal hemoglobinuria in a girl with hemolysis and “hematuria”. Pediatr Nephrol. 2004;19: 1177–1179. pmid:15278422
- 32. O’Connor JPB, Tofts PS, Miles KA, Parkes LM, Thompson G, Jackson A. Dynamic contrast-enhanced imaging techniques: CT and MRI. Br J Radiol. 2011;84: 112–120. pmid:20959377
- 33. Evelhoch JL. Key factors in the acquisition of contrast kinetic data for oncology. J Magn Reson Imaging. 1999;10: 254–259. pmid:10508284
- 34. Turkbey B, Kobayashi H, Ogawa M, Bernardo M, Choyke PL. Imaging of Tumor Angiogenesis: Functional or Targeted? Am J Roentgenol. 2009;193: 304–313. pmid:19620425
- 35. Barrett T, Brechbiel M, Bernardo M, Choyke PL. MRI of Tumor Angiogenesis. J Magn Reson Imaging. 2007;26: 235–249. pmid:17623889
- 36. Haroon HA, Buckley DL, Patankar TA, Dow GR, Rutherford SA, Balériaux D, et al. A comparison of Ktrans measurements obtained with conventional and first pass pharmacokinetic models in human gliomas. J Magn Reson Imaging. 2004;19: 527–536. pmid:15112301