Konventionelle und molekularpathologische Untersuchungen während und nach klinischer Herztransplantation View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1997-01

AUTHORS

Ph. A. Schnabel, R. Lange, K. Amann, T. M. Bingoid, H. Jakob, M. Kampmann, A. Koch, B. Koch, A. Magener, H. Mehmanesh, Z. Niemir, F. -U. Sack, M. Schmid, A. Schmiedl, R. Waldherr, R. Zimmermann, S. Hagl, H. F. Otto

ABSTRACT

The most important diagnostic questions following HTx relate to graft failure, acute rejection, infections and chronic rejection. Right ventricular biopsies were taken during and following HTx at different time points: a) before cardioplegia, b) immediately following cardioplegia with Custodiol® and explantation, c) before implantation, d) after 30 and/or 60 min reperfusion, e) weekly during the first postoperative month and then after longer time intervals. After fixation of the probes in formalin and rapid embedding in paraffin, histopathology of the myocardium was evaluated, immunocytochemical analyses followed. Applying fixation in buffered glutar- and paraformaldehyde and embedding in epon-araldite, myocardial ultrastructural preservation was evaluated. Samples rapidly frozen in liquid nitrogen were taken for immunocytochemical and/or molecular investigations. By means of conventional light and electron microscopy, alterations of the composition of the myocardium (interstitial infiltrates, fibrosis), of cardiac myocytes (cellular edema, hypertrophy), and of endothelia (damage of cell organelles, cellular edema) were evaluated qualitatively and morphometrically. Molecular alterations underlying inflammation (acute myocardial and/or vascular rejections, infections) or complex processes such as chronic rejection were investigated on the protein level by means of immunocytochemistry and on the DNA or mRNA level by means of the polymerase chain reaction (PCR) or in situ hybridization.Qualitatively and morphometrically, light microscopy could not detect differences between biopsies taken after explantation and those before implantation following 4 hours of ischemia. Ultrastructural stereology was able to demonstrate that during HTx capillary endothelia were more susceptible to ischemia/reperfusion than cardiac myocytes. Single quantitative ultra-structural parameters for structural protection showed statistically significant correlations with the duration of ischemia. Different parameters correlated with the number and severity of myocardial or vascular rejections during the first year after HTx. Infections with the cytomegalovirus were detected by means of immunocytochemistry and in situ PCR. Altered expression of cytokines and growth factors was demonstrated by means of immunocytochemistry and in situ hybridization.Thus, different questions following HTx can only be answered by application of corresponding conventional and/or molecular methods, provided that adaequately fixed samples are prepared. More... »

PAGES

7-22

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/bf03044660

DOI

http://dx.doi.org/10.1007/bf03044660

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1048609014


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10 schema:description The most important diagnostic questions following HTx relate to graft failure, acute rejection, infections and chronic rejection. Right ventricular biopsies were taken during and following HTx at different time points: a) before cardioplegia, b) immediately following cardioplegia with Custodiol® and explantation, c) before implantation, d) after 30 and/or 60 min reperfusion, e) weekly during the first postoperative month and then after longer time intervals. After fixation of the probes in formalin and rapid embedding in paraffin, histopathology of the myocardium was evaluated, immunocytochemical analyses followed. Applying fixation in buffered glutar- and paraformaldehyde and embedding in epon-araldite, myocardial ultrastructural preservation was evaluated. Samples rapidly frozen in liquid nitrogen were taken for immunocytochemical and/or molecular investigations. By means of conventional light and electron microscopy, alterations of the composition of the myocardium (interstitial infiltrates, fibrosis), of cardiac myocytes (cellular edema, hypertrophy), and of endothelia (damage of cell organelles, cellular edema) were evaluated qualitatively and morphometrically. Molecular alterations underlying inflammation (acute myocardial and/or vascular rejections, infections) or complex processes such as chronic rejection were investigated on the protein level by means of immunocytochemistry and on the DNA or mRNA level by means of the polymerase chain reaction (PCR) or in situ hybridization.Qualitatively and morphometrically, light microscopy could not detect differences between biopsies taken after explantation and those before implantation following 4 hours of ischemia. Ultrastructural stereology was able to demonstrate that during HTx capillary endothelia were more susceptible to ischemia/reperfusion than cardiac myocytes. Single quantitative ultra-structural parameters for structural protection showed statistically significant correlations with the duration of ischemia. Different parameters correlated with the number and severity of myocardial or vascular rejections during the first year after HTx. Infections with the cytomegalovirus were detected by means of immunocytochemistry and in situ PCR. Altered expression of cytokines and growth factors was demonstrated by means of immunocytochemistry and in situ hybridization.Thus, different questions following HTx can only be answered by application of corresponding conventional and/or molecular methods, provided that adaequately fixed samples are prepared.
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16 schema:keywords Custodiol
17 DNA
18 Epon-Araldite
19 HTx
20 Untersuchungen
21 acute rejection
22 alterations
23 altered expression
24 analysis
25 applications
26 biopsy
27 capillary endothelium
28 cardiac myocytes
29 cardioplegia
30 chain reaction
31 chronic rejection
32 complex process
33 composition
34 conventional light
35 correlation
36 cytokines
37 cytomegalovirus
38 diagnostic questions
39 differences
40 different parameters
41 different questions
42 different time points
43 duration
44 duration of ischemia
45 electron microscopy
46 embedding
47 endothelium
48 explantation
49 expression
50 factors
51 failure
52 first postoperative month
53 first year
54 fixation
55 formalin
56 graft failure
57 growth factor
58 histopathology
59 hours
60 hours of ischemia
61 hybridization
62 immunocytochemical analysis
63 immunocytochemistry
64 implantation
65 important diagnostic questions
66 infection
67 inflammation
68 interval
69 investigation
70 ischemia
71 ischemia/reperfusion
72 levels
73 light
74 light microscopy
75 liquid nitrogen
76 long time intervals
77 mRNA levels
78 means
79 means of immunocytochemistry
80 method
81 microscopy
82 min reperfusion
83 molecular alterations
84 molecular investigations
85 molecular methods
86 months
87 myocardium
88 myocytes
89 nitrogen
90 number
91 paraffin
92 paraformaldehyde
93 parameters
94 point
95 polymerase chain reaction
96 postoperative month
97 preservation
98 probe
99 process
100 protection
101 protein levels
102 questions
103 rapid embedding
104 reaction
105 rejection
106 relate
107 reperfusion
108 right ventricular biopsies
109 samples
110 severity
111 significant correlation
112 situ hybridization
113 situ polymerase chain reaction
114 stereology
115 structural protection
116 time interval
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118 ultrastructural preservation
119 ultrastructural stereology
120 vascular rejection
121 ventricular biopsies
122 weekly
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