Pompe's disease - RePub - Erasmus Universiteit Rotterdam

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Cardiac remodeling and contractile function in KO-mice Biochemistry and morphology At the conclusion of each experiment, the heart was excised and the atria were removed. The right and left ventricle were separated, weighed and freeze dried for the determination of dry weights and, in the fi rst part of the study, for determination of acid α-glucosidase activity and glycogen content. In addition, samples were also obtained from the musculus quadriceps femoris and the liver to measure acid α-glucosidase activity and glycogen content. The atria and another sample from the musculus quadriceps femoris were fi xed in glutaraldehyde (4%), embedded in glycolmethacrylate and histological sections of 4 µm were stained with haematoxylin azofl oxin and periodic acid schiff. Tail DNA was extracted to verify the genotype by PCR (5). Atria, muscle and liver samples were homogenized in phosphate buffered saline, using an Ultra Turax (TP18-20, 20000 UPM; 170W; see reference 5). Large debris was removed after centrifugation at 10,000 g for 15 min. The supernatant was sonicated on ice for two times 10 s at an amplitude of 10 µm, and analysed for total protein concentration with the Bichionic Acid Protein Assay (Pierce, Rockford, IL) and acid α-glucosidase activity with 4MU (4-methylumbelliferyl-α-D-glucopyranoside) as substrate at a pH of 4. The glycogen content was measured after dialysis of the sample to remove cytoplasmic glucose, as previously described (5,28). Data analysis Echocardiographic data were collected on an optical disk and stored for off-line analysis. Left ventricular (LV) diameters at end-diastole (EDD) and end-systole (ESD) as well as the ED and ES thickness of the LV posterior wall were measured from the M-mode images using Clemex Vision software package (Clemex Technologies, Quebec, Canada). Five consecutive beats were analyzed in triplicate and averaged. LV absolute shortening (EDD - ESD) and fractional shortening ((EDD - ESD)/EDD*100%) were calculated. Hemodynamic data were recorded and digitized using an on-line 4-channel data acquisition program (ATCODAS, Dataq Instruments, Akron, OH) for post-acquisition off-line analysis with a program written in MATLAB (Mathworks, Natick, MA). Fifteen consecutive beats were selected for determination of heart rate, LV peak systolic and end-diastolic pressures, diastolic aortic pressure and the maximum rate of rise of LV pressure (LV dP/dt max ). In addition, the time constant (τ) of LV pressure decay, an index of early LV relaxation, was computed as described earlier (11). Statistical analysis of anatomical and echocardographic data was performed using unpaired t-test or McNemar test as appropriate. Analysis of hemodynamic data was performed using two way analysis of variance followed by post-hoc testing with paired and unpaired t-test or signed rank test and McNemar test, as appropriate. Statistical signifi cance was accepted when P
Chapter 2 Macroscopy The wet weight of the heart (HW) was on average 44% higher in KO compared to WT mice (P
Page 2 and 3: Pompe’s disease The mouse as mode
Page 4 and 5: Pompe’s disease The mouse as mode
Page 6: Contents Chapter 1 Introduction 11
Page 10: 1Introduction
Page 13 and 14: Chapter 1 In lysosomal storage diso
Page 15 and 16: Chapter 1 Network are associated wi
Page 17 and 18: Chapter 1 the indirect approach can
Page 19 and 20: Chapter 1 Diagnosis Clinical Diagno
Page 21 and 22: Chapter 1 Fig. 3. Targeting vectors
Page 23 and 24: Chapter 1 muscles in approximately
Page 25 and 26: Chapter 1 1.7 Scope The studies des
Page 27 and 28: Chapter 1 47. Rudolph, C., et al.,
Page 29 and 30: Chapter 1 112. Dorling, P.R., J.M.
Page 32: 2 Cardiac Remodeling and Contractil
Page 35: Chapter 2 Experimental groups The a
Page 39 and 40: Chapter 2 Pooling of all individual
Page 41 and 42: Chapter 2 Hemodynamics during isofl
Page 43 and 44: Chapter 2 Discussion The major fi n
Page 45 and 46: Chapter 2 References 1. Bharati S,
Page 48: 3 Long Term Intravenous Treatment o
Page 51 and 52: Chapter 3 gene, we have focused on
Page 53 and 54: Chapter 3 pattern of alpha-glucosid
Page 55 and 56: Chapter 3 Alpha-glucosidase activit
Page 57 and 58: Chapter 3 and 4, respectively. The
Page 59 and 60: Chapter 3 For patient 1 the mental
Page 61 and 62: Chapter 3 R.M. Koppenol, T. de Vrie
Page 64: 4 Morphological Changes in Muscle T
Page 67 and 68: Chapter 4 Therapeutic regimen Pharm
Page 69 and 70: Chapter 4 Fig. 2. Pre treatment var
Page 71 and 72: Chapter 4 Fig. 4. Effect of ERT on
Page 73 and 74: Chapter 4 Table 2. Patients scores
Page 75 and 76: Chapter 4 enzyme. Moreover, when th
Page 78: 5Hearing loss in Infantile Pompe’
Page 81 and 82: Chapter 5 Methods Patients Seven pa
Page 83 and 84: Chapter 5 revealed a hearing loss o
Page 85 and 86: Chapter 5 Fig. 3. Threshold levels
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Chapter 5 Discussion Hearing loss i
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6 Both Low and High Uptake Forms of
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Chapter 6 healthy volunteers, we st
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Chapter 6 was extracted from human
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Chapter 6 from bovine testes (bAGLU
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Chapter 6 Fig. 5. Immunocytochemica
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Chapter 6 glucuronidase, and is obv
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Chapter 6 van Deel MSc for technica
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7Discussion
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Chapter 7 Cardiac function The mous
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Chapter 7 After the fi rst three mo
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Chapter 7 in 2004. This is 40 years
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Chapter 7 the skeletal muscle. The
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Chapter 7 46. Baudhuin, P., H.G. He
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age are ventilator dependent and wh
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om de morfologie van de spieren te
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naar de ziekte van Pompe bij kinder
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Publications Cardiac remodeling and
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Dankwoord Of ‘Mice and men’ zou
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Dit proefschrift had er niet zo uit
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Pompe's disease - RePub - Erasmus Universiteit Rotterdam

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