Pathology of endomyocardial biopsy

Endomyocardial biopsy (EMB) is routinely performed for cardiac transplant monitoring. Moreover, it can be a useful tool in the diagnostic work-up of myocarditis, cardiomyopathies, drug toxicity, unexplained arrhythmias, heart involvement in systemic disease, and to diagnose cardiac masses. Pathologic evaluation of EMB represents the gold standard in specific disorders, including myocarditis and cardiac involvement by amyloidosis and sarcoidosis. To increase the diagnostic yield of the procedure, multiple myocardial samples should be obtained, and processed in a fully-equipped laboratory, with access to immunohistochemical/histochemical analyses, molecular tests and transmission electron microscopy. We here review the main diagnostic features of myocarditis, cardiomyopathies and cardiac tumours on EMB, specifying the ancillary techniques required to reach the diagnosis in each specific cardiac disease.     

Three-dimensional reconstruction of abnormal intramural coronary arteries in human cardiac allograft biopsies

A novel abnormality of intramural coronary arteries has been recently described in allograft cardiac biopsies. Three-dimensional (3-D) models of these abnormal arteries have now been constructed from serial histological sections of diagnostic post-transplant endomyocardial biopsies. These revealed that there was an interlacing meshwork of longitudinal smooth muscle bundles in abnormal arteries. In addition, the lumen and external surface of these arteries were irregular. This contrasted with reconstructions of normal control arteries in the same sections, which were smooth and straight. This elucidation of an unusual abnormality suggests that 3-D reconstruction of arteries in forceps biopsies may be a useful technique. © 1997 John Wiley & Sons, Ltd.     

Arterial lesions associated with medial disorganization and fibrosis in endomyocardial biopsies from human cardiac allografts

We describe distinctive arterial lesions in endomyocardial biopsies from patients with human cardiac allografts. The lesions affected principally the media of small arteries and consisted of misorientation of smooth muscle cells and fibrosis. This remodelling was most prevalent in the subadventitial zone, but sometimes extended to involve the full thickness of the media. In the most extreme cases medial smooth muscle cells ran parallel to the long axis of the vessel and were segregated into small bundles and single cells separated by collagen which merged with the adventitial fibrosis. The intima was always normal. Abnormal arteries were present in 16% of 603 consecutive biopsies from 44 patients, and 39% of lesions occurred in 16% of patients. No lesions were found in endomyocardial biopsies from 25 non-transplanted patients, nor in mucosal biopsies from both transplanted and non-transplanted patients, confirming that the appearances were not due to biopsy artefact. There was early arterial remodelling in biopsies within two weeks of transplantation and none of the stages resembled vascular rejection. Fifty per cent of biopsies from some patients contained arterial lesions, suggesting that in susceptible patients they are common.     

Myocardial fibrosis assessment by semiquantitative, point-counting and computer-based methods in patients with heart muscle disease: a comparative study

AIMS: No study has directly compared different histomorphometric methods of quantification of myocardial fibrosis. Therefore we compared the results of semiquantitative, point-counting and computer-based methods in the assessement of myocardial fibrosis in a consecutive series of endomyocardial biopsy samples from patients with heart muscle disease. METHODS AND RESULTS: Histological samples (at least three per patient) were obtained by endomyocardial biopsy from 11 patients with focal myocarditis and from 24 ambulatory patients with idiopathic dilated cardiomyopathy, or during surgery in 10 patients who underwent partial left ventriculectomy. Samples were cut and stained with Masson-trichrome for better contrast. From each sample, a representative field was digitized, and the amount of fibrosis was assessed by semiquantitative scoring, by point-counting, and by computer-based software. Semiquantitative scoring correlated with both point-counting (Spearman's r = 0.69, P < 0.0001) and computer-based (Spearman's r = 0.83, P < 0.0001) methods. There was also a good correlation between point-counting and computer-based methods (r = 0.71, P < 0.0001). However, when compared with the point-counting method, the computer-based method overestimated percent fibrosis by 3.0 +/- 6.7% (P = 0.004). This overestimation correlated with the mean percent fibrosis (r = 0.38, P = 0.014). CONCLUSIONS: Our data show good correlations between the three methods of myocardial fibrosis assessment. However, systematic differences between them emphasize that this should be taken into consideration when comparing results of the studies using different methods of fibrosis assessment.     

Endomyocardial biopsy pathology in insulin-dependent diabetic patients with abnormal ventricular function

We have previously shown impaired ventricular function in asymptomatic middle-aged type 1 (insulin-dependent) diabetic patients who had no evidence of coronary artery disease. The diabetic patients had normal coronary angiograms but reduced ventricular ejection fraction on exercise. To examine the possible contribution of small vessel disease to this functional abnormality, we compared endomyocardial biopsies from seven symptom-free type 1 diabetic patients with biopsies from seven age- and sex-matched non-diabetic subjects. Interstitial fibrosis was present in three diabetic patients, arteriolar hyalinization in three patients and arteriolar thickening was observed in five patients. Morphometry performed on electron micrographs showed no significant difference in the thickness of the capillary basal lamina between diabetics and controls. While the functional significance of the abnormalities on light microscopy is unclear, our findings indicate that the abnormality of cardiac function described in diabetes is not associated with thickening of the myocardial capillary basal lamina.     

Association between enterovirus endomyocardial infection and late severe cardiac events in some adult patients receiving heart transplants

Enteroviruses and other cardiotropic viruses have been associated with the development of late severe adverse cardiac events in infants receiving heart transplants. However, the source and the chronology of cardiac allograft infection by an enterovirus in patients receiving heart transplants remain unknown. Using RT-PCR and immunohistochemistry assays, endomyocardial tissue samples of 30 adult patients were tested to detect the presence of specific enterovirus 5' non-coding (5'NC) sequences and of VP1 capsid protein, and this at the time of cardiac transplantation and at the 12-month biopsy for graft rejection control. Moreover, the endomyocardial detection of genomic sequences of enteroviruses, Epstein-Barr virus, herpes simplex virus, cytomegalovirus (CMV), varicella-zoster virus, adenoviruses, and parvovirus B19 was carried out by RT-PCR and polymerase chain reaction (PCR) assays at the time of late severe cardiac events. Enterovirus RNA and VP1 antigen were both detected in 4 (13%) of 30 patients at the time of the 12-month biopsy for graft rejection control, whereas no enterovirus component was detected in the explanted and implanted heart tissues taken from these 4 patients at the time of transplantation. At the time when severe cardiac events were developed, within 3 months after the positive enterovirus cardiac detection, these four patients demonstrated the presence of endomyocardial enterovirus RNA sequences whereas they were tested negative for the endomyocardial detection of genomic sequences from DNA viruses (except for CMV in two cases), and for a significant level of pp65 CMV antigenemia. Taken together, these findings indicate that enteroviruses could be acquired as a new endomyocardial infection within 12 months after transplantation in adults receiving heart transplants, and suggest that this infection might be an etiological cause for unexplained late severe adverse cardiac events in the heart-transplantated adults.     

Acute vascular humoral rejection in a sensitized cardiac graft recipient: diagnostic value of C4d immunofluorescence

A 37-year-old female patient had a cardiac transplantation for dilated cardiomyopathy. She was sensitized by two pregnancies showing anti-human leukocyte antigen I and II antibodies. The pretransplantation crossmatch was negative, but she developed acute humoral rejection characterized by vascular C4d deposits, arteriolitis, and intravascular leukocyte accumulation and adhesion in venules. Although C4d deposits disappeared in 4 weeks, she had persistent endothelial cell activation (endothelial expression of ELAM-1, VCAM-1, or human leukocyte antigen class II) throughout the 6 months of follow-up. Although she received intensive immunosuppression, she presented three episodes of acute cellular rejection during that period of time. This case shows that C4d deposits represent a sensitive marker of acute humoral rejection in cardiac transplantation. Therefore, C4d immunofluorescence should be more frequently assessed in endomyocardial biopsies.     

An examination of the variation in timed endometrial biopsies

Endometrial biopsies were obtained from fertile women, aged20–40 years, with regular cycles of 25–35 days.Chronological dating of the material was carried out by determinationof the luteinizing hormone (LH) peak by daily LH assay. Tissuewas taken from three or four sites along the biopsy and processedfor electron microscopy. Half micron thick sections were analysedby light microscopy and a morphometric examination of the glandularepithelium carried out. Nuclear size was assessed using an unbiasedmeasuring technique and the volume density of the nucleus inthe cell was also measured. A series of 12 women was examined,four at each of days LH+2,4 and 5. An attempt was made to controla variety of factors which contribute to the variability inthe histological dating of the endometrial biopsy. In doingso it was possible to analyse the relative contributions ofeach sampling level to the overall variance. In contrast toearlier studies there is a relatively small inter-subject variationwith a correspondingly large within-biopsy variation. Theseresults may indicate that the cellular events in the glandularepithelium, between ovulation and the middle of the luteal phase,are precisely regulated.     

Ultrastructural Findings in Cardiac Transplant Recipients

Ultrastructural findings in 350 endomyocardial biopsy specimens and 59 autopsy or explanted hearts from cardiac transplant recipients are reviewed. Myocyte degeneration can be readily distinguished from necrosis by the technique described. Vascular changes of endothelial activation, endothelial cell damage, and basement membrane reduplication can be readily identified. In addition, the myofilament composition of ischemic hearts in patients with allograft coronary artery disease is distinctive: There is a disproportionate loss of actin over myosin, giving a coarse appearance to the myofilaments. These changes are useful in further defining the morphologic features associated with rejection and ischemia in cardiac transplant recipients.     

Perforin and granzyme B as markers for acute rejection in heart transplantation.

Histological analysis of endomyocardial biopsies (EMB) is regarded as the most satisfactory technique for monitoring crisis of rejection in heart transplanted patients. In this study, 42 biopsies from 14 patients who underwent heart transplantation were examined. Three patients did not present any rejection crisis at the date of the biopsy analysis, six were examined during an early rejection crisis (day 7-70 post-graft), and five were examined during a late rejection crisis (day 74-960 post-graft). Since granzyme B and perforin are proteins associated with cell lysis histological grading and cell phenotype analysis, in situ hybridization using granzyme B and perforin [35S]RNA probes was performed on 30 EMB to characterize the cytolytic activation of heart infiltrating cells. Our data suggest that granzyme B and perforin could be used as predictive markers for acute rejection in patients with early rejection crisis. Their detection might be an indication to administrate corticoids to resolve an acute rejection crisis. In contrast, their absence in patients with late rejection crisis appears as a good prognostic factor for the outcome of rejection and raises the question of the necessity to treat such patients with additional corticoid treatment.     

Human cardiac transplants diagnosis of rejection by endomyocardial biopsy causes of death (about 30 autopsies)

Summary 1,000 endomyocardial biopsies performed in 110 patients treated by cardiac graft were reviewed. These biopsies permitted early detection of acute rejection after cyclosporin treatment and a good appreciation of its intensity. By this method, almost all rejection episodes were resolved after adequate treatment. Chronic rejection was diagnosed by arteriography used in vivo or in cardiac transplants removed by surgery or necropsy. Rejection provoked an obliterative fibrous endarteritis often complicated by atherosclerosis and its ischaemic consequences.34 autopsies were performed in patients dead at a variable time after cardiac or cardio-pulmonary transplantation. In early death (14 cases), graft failure and systemic disorders were observed. Acute and chronic rejection was noted less frequently (9 cases). Systemic infections (10 cases) occured either early (post-surgical complications) or late (bacterial, fungal and parasitic lesions). In one case, death was due to a contemporaneous bladder carcinoma.The complications of cyclosporin treatment are briefly discussed.An antifungic agent (cyclic peptid) isolated from Trichoderma polysporum, with probableAn antifungic agent (cyclic peptid) isolated from Trichoderma polysporum, with probable     

Inflammation in myocardial disease: From myocarditis to dilated cardiomyopathy

Dilated cardiomyopathy (DCM) is a heterogeneous group of myocardial diseases clinically defined by the presence of left ventricular dilatation and contractile dysfunction. Among various causes of DCM, a progression from viral myocarditis to DCM has long been hypothesized. Supporting this possibility, studies by endomyocardial biopsy, the only method to obtain a definite diagnosis of myocarditis at present, have provided evidence of inflammation in the myocardium in DCM patients. A number of experimental studies have elucidated a cell‐mediated autoimmune mechanism triggered by viral infection in the progression of myocarditis to DCM. In addition, the important role of inflammation in the pathogenesis of heart failure has been recognized, and many terms including myocarditis, inflammatory cardiomyopathy, and inflammatory DCM have been used for myocardial diseases associated with inflammation. This review discusses the pathophysiology of inflammation in the myocardium, and refers to diagnosis and treatment based on these concepts.     

Ultrastructural Aspect of Myocarditis: Its Relevance for the Diagnosis

The Dallas consensus was used to reveal active or borderline inflammatory loci by light microscopy (LM). When lympho-cyte-cardiocyte interaction was observed by electron microscopy (EM), the deleterious or dormant pattern of inflammatory process was recognized. The first was determined by lymphocytes that adhered to cardiocytes, next to necrotic cardiocytes or admixed with debris. The second was marked by scattered lymphocytes between preserved cardiocytes and the absence of lymphocytes adhered to cardiocytes and necrotic cardiocytes. The deleterious pattern of the inflammatory process (EM) commonly supplemented the active appearance of inflammatory loci (LM). In contrast, the borderline outlook of the LM completed either the deleterious or dormant pattern of the EM. This discrepancy was related to the restricted resolution of LM, which might hide the actual stage of the disease. The diagnosis of myocarditis was founded on mutual LM and EM observations. The active or borderline appearance of LM of the deleterious pattern (EM) was considered indicative for the active stage of myocarditis. The borderline outlook of the LM of the dormant pattern of the EM was admitted to indicate either the healing phase of the disease with lymphocytes still lagging behind, or a latent phase of the ongoing myocarditis, according to the patient's hemodynamic status.     

The pathological implications of heart transplantation: Experience with 50 cases in a single center

Heart transplantation started in Japan in 1999. Since then, 50 transplants have been performed at our center. We performed histopathological analyses of the 50 explanted hearts and the post‐transplant biopsy specimens. The median age of recipients was 39 years. The primary diseases before transplant were idiopathic dilated cardiomyopathy in 33 patients (66%), hypertrophic cardiomyopathy in seven (14%), restrictive cardiomyopathy in one, arrhythmogenic right ventricular cardiomyopathy in one, and secondary cardiomyopathy in eight (16%). Before transplantation, 47 patients (94%) had left ventricular assist devices. No severe cardiovascular failure due to allograft rejection occurred. The post‐transplant survival rate was 97.6% at 1 year and 93.1% at 10 years. One recipient was lost to sepsis from myelodysplastic syndrome in the fourth year, one died of multiple organ failure and peritonitis 8 months after transplant. Another patient died of recurrent post‐transplant lymphoproliferative disorders (PTLD). Mild cardiac dysfunction occurred in seven recipients in the early postoperative period. Moderate acute cellular rejection occurred in six patients (12%), and antibody‐mediated rejection occurred in three (6%). The number of heart transplants performed in Japan is very small. However, the outstanding 10‐year survival rate is due to donor evaluation and post‐transplant care resulting in low grade rejection. Pathological evaluation has also greatly contributed to the results.     

Human cardiac transplantation--evaluation of morphological changes in serial endomyocardial biopsies

From May 1981 through July 1984 a total of 29 human allogenic orthotopic cardiac transplants were performed in Munich. The first two patients initially received conventional immunosuppressive treatment for 79 and 27 days, respectively; then treatment was continued with cyclosporine. All subsequent 27 patients received only cyclosporine treatment. Seventeen of the cardiac recipients are currently alive. Three of the recipients who died succumbed to immunological rejection. Twenty-five cardiac grafts were controlled by 355 sequential biopsy procedures, which yielded 1158 endomyocardial specimens for histological examination. The morphological findings and changes observed in the endomyocardium were analyzed. The interpretation of these findings and difficulties encountered in their interpretation are discussed. Special attention is attributed to findings possibly associated with the cyclosporine treatment.     

A neural network approach to the biopsy diagnosis of early acute renal transplant rejection

AIMS: To develop and test a neural network to assist in the histological diagnosis of early acute renal allograft rejection. METHODS AND RESULTS: We used three sets of biopsies to train and test the network: 100 'routine' biopsies from Leicester; 21 selected difficult biopsies which had already been evaluated by most of the renal transplant pathologists in the UK, in a study of the Banff classification of allograft pathology and 25 cases which had been classified as 'borderline' according to the Banff classification in a review of transplant biopsies from Oxford. The correct diagnosis for each biopsy was defined by careful retrospective clinical review. Biopsies where this review did not provide a clear diagnosis were excluded. Each biopsy was graded for 12 histological features and the data was entered into a simple single layer perception network, designed using the MATLAB neural network toolbox. Results were compared with logistic regression using the same data, and with 'conventional' histological diagnosis. If the network was trained only with the 100 'routine' cases, its performance with either of the other sets was poor. However, if either of the 'difficult' sets was added to the training group, testing with the other 'difficult' group improved dramatically; 19 of the 21 'Banff' study cases were diagnosed correctly. This was achieved using observations made by a trainee pathologist. The result is better than was achieved by any of the many experienced pathologists who had previously seen these biopsies (maximum 18/21 correct), and is considerably better than that achieved by using logistic regression with the same data. CONCLUSION: A neural network can provide a considerable improvement in the diagnosis of early acute allograft rejection, though further development work will be needed before this becomes a routine diagnostic tool. The selection of cases used to train the network is crucial to the quality of its performance. There is scope to improve the system further by incorporating clinical information. Other related areas where this approach is likely to be of value are discussed.     

An appraisal of the value of the bone marrow biopsy in the assessment of proliferative lesions of the bone marrow

The diagnostic value of the bone marrow needle biopsy has proved impressive in a variety of disorders. As a complementary procedure to the aspiration smear it adds an invaluable dimension to the examination of haematopoietic tissue. The procedure is easily learned and safe and should be utilized routinely in haematological practice. The usefulness of the bone marrow biopsy is examined in assessing proliferative lesions of the bone marrow.     

供者脾细胞对移植心脏免疫耐受的诱导

目的探讨供者脾细胞对同种异体心脏移植免疫耐受的诱导效果 ,为抗排斥反应治疗提供依据。方法分别采用供者脾细胞 (SPC)和环磷酰胺 (CP)预处理移植受者 ,然后行大鼠异位心脏移植术 ,根据实验分组对移植心脏存活情况进行观察。结果对照组、CP组和 SPC组移植心脏的存活时间分别为 7.2 1± 2 .5 6 d、9.78± 2 .5 5 d和 15 .14± 8.5 6 d,经统计学处理后证实 ,3组移植心脏的存活时间有显著性差异 (P<0 .0 5 )。结论供者脾细胞预处理移植受体 ,可以明显地延长大鼠同种异体心脏移植的存活时间。     

A morphometric assessment of transitional mucosa in the colon

Quantitative morphometric analysis was used in 10 resection specimens to assess so-called transitional mucosa immediately adjacent to colorectal carcinoma. Eleven nuclear and cellular variables were measured from the malignant epithelial area and from zones of increasing distance (1 cm) from the lesion. In addition, mean mucosal height was assessed for each zone. Morphometrical differences between the mucosa immediately adjacent to the malignant epithelium and that taken at some distance from it were determined by Mann-Whitney U tests. Transitional mucosa showed increased mucosal height but no nuclear differences from normal mucosa. Other work has shown that there are nuclear morphometric differences associated with premalignant conditions in the colon. Thus, the suggestion that transitional mucosa represents early neoplastic change cannot be supported.     

Remodeling of fetoplacental arteries in rats due to chronic hypoxia

The increased fetoplacental vascular resistance due to chronic hypoxia cannot be explained by simple hypoxic vasoconstriction, as it sustains to some degree after recovery in normobaric environment. To verify a hypothesis that fetoplacental arteries undergo remodeling of their walls similar to remodeling of pulmonary arteries in hypoxic pulmonary hypertension, we used a model of the chronically hypoxic rat placenta. Han Wistar pregnant rats were exposed to 14-day hypoxia (10% of oxygen) during the 6th to 19th day of pregnancy. Chronic hypoxia elicited in both intraplacental (prelabyrinthine) and chorionic plate (insertion) arteries significant narrowing of their lumina. Irregular thickening of their adventitia due to an increase in collagen fibers as well as ground substance was observed; reticular fibers were fragmented. Because of remodeling of fetoplacental arteries, a model of chronically hypoxic rat placenta could simulate human preplacental hypoxia and consequent effects.