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To determine parameters of predictive value in CML, a retrospective clinico-pathological study was performed. This included laboratory data and (pretreatment) bone marrow biopsies of 120 patients with a monotherapy by busulfan (BU) and 50 patients with interferon-alpha 2b (IFN)treatment. Median survival in the BU group was 39 months and in the IFN-treated patients 65 months. Morphological features (CD61-positive megakaryocytes, argyrophilic fibres, pseudo-Gaucher cells) were evaluated by morphometry. Additionally, we measured the incidence of apoptosis (in situ end-labelling technique) and the expression of the proliferating cell nuclear antigen (PCNA). The ratio between the proliferative and apoptotic cell fraction was coined leukaemia turnover index (LTI). In order to estimate the impact of clinical and various morphological as well as dynamic features of prognostic significance, a multivariate analysis was carried out using the classification and regression tree approach (CART). Discrimination of single disease parameters revealed that fibrosis remained the most significant variable for survival in both therapeutic groups. Indicators of myeloid metaplasia such as occurrence of erythro-normoblasts and/or splenomegaly were important clinical parameters for prognosis. Inclusion of morphological as well as dynamic disease features in risk classification resulted in a substantial improvement of prognostic efficiency compared to other predictive scores which could be demonstrated by means of ROC-analysis.

Morphometric analysis of cell-nucleus size was undertaken in 300 cases of malignant non-Hodgkin lymphomas. The results were compared with histological findings and survival time. It was found that the histological lymphoma groups differ significantly only occasionally with regard to nucleus size. Within the groups the values have such wide scatter that in the individual case it is not possible to assign it on morphometric data. This scatter mirrors the difficulties encountered in histological classification which - as the objective data indicate - are due to numerous transitions between different types of lymphoma. Analysis of survival data shows that nucleus size is of considerable prognostic value, if certain exceptions such as lymphoblastic lymphoma are taken into account. The results favour tripartite division of malignant lymphomas for prognostic purposes.

Aims Bone marrow histopathology reveals a striking heterogeneity at diagnosis of Philadelphia chromosome positive (Ph1+) chronic myelogenous leukaemia (CML). Based on semiquantitative evaluations of the number of megakaryocytes and the content of fibres, various histological subtypes have been postulated. However, little information exists on whether these groups represent stable categories of the different classification systems and whether therapeutic regimes exert any influence on the putative shift of histological patterns. Methods and results A retrospective clinicopathological study was performed on 396 bone marrow biopsies derived from 173 patients. There were at least two representative trephines taken at diagnosis and at median intervals of 16 months. Processing of the specimens involved immunostaining with CD61 (megakaryopoiesis) and Ret40f (erythropoiesis) and Gomori's silver impregnation technique. Based on morphometric analysis and in accordance with the general appearance of bone marrow histology three different histological subtypes were distinguished. These consisted of a granulocytic (51 patients), a predominantly megakaryocytic (73 patients) and a myelofibrotic pattern (49 patients). Follow-up biopsies revealed that a significant transition of histological groups occurred and that, independently of treatment modalities, the myelofibrotic category was associated with an unfavourable prognosis. Of the 124 patients without myelofibrosis at onset, 42% later transformed into the myelofibrotic subtype. However, these patients showed no prevalence of either a pre-existing granulocytic or megakaryocytic growth. Myelofibrotic changes were significantly associated with interferon (IFN) and busulfan (BU) therapy. On the other hand, a transition of a myelofibrotic into a nonfibrotic subtype was detectable in 17 of the 49 patients under study and related to hydroxyurea (HU) treatment. Conclusions Histological classification systems of bone marrow features in CML do not represent stable patterns, but may be significantly altered by therapy, in particular IFN and HU.

A clinicopathological study was conducted on 351 bone marrow trephine biopsies derived from 124 patients with chronic myeloid leukemia (CML) at standardized endpoints before and after allogeneic bone marrow transplantation (BMT). The purpose was to investigate quantitative changes of the nucleated erythroid precursor cell population and other associated features such as resident bone marrow macrophages and myelofibrosis and to elucidate their relevance on engraftment parameters. Monoclonal antibodies were applied for the identification of erythroid precursors and the labeling of mature macrophages; argyrophilic (reticulin-collagen) fibers were demonstrated by a silver impregnation technique. Following morphometric analysis of the pregraft bone marrow specimens statistical evaluation was in line with an adverse correlation between early to moderate reticulin fibrosis and amount of erythropoiesis. Moreover, a significant relationship was calculable between numbers of erythroid precursors and CD68⁺ macrophages. After myelo-ablative therapy and BMT a pronounced decrease in cellularity and in the quantity of erythropoiesis was found. Comparable with the pregraft samples, a significant association between erythroid precursors and macrophages could be determined in the regenerating donor bone marrow. A pretransplant relevant reduction of the red cell lineage and a manifest (reticulin) myelofibrosis indicating an advanced stage of disease were accompanied by a significant delay to reach transfusion independence. This result was further supported by comparable findings in trephine biopsies performed in the early post-transplant period (second month after BMT). Corresponding examinations revealed an enhancement of fiber density and a decrease in erythropoiesis in those patients who did not conform with the usually accepted criteria for successful engraftment. In conclusion, compelling evidence has been produced that a significantly reduced amount of erythroid precursors, which is usually associated with myelofibrosis in the pretransplant bone marrow, exerts an impairment to undisturbed hematopoietic reconstitution. Moreover, a close spatial and numerical relationship between the erythroid lineage and resident (mature) macrophages is observable, in particular in the state of regeneration after BMT.

Striking morphological similarities exist between T-cell-rich B-cell lymphoma and lymphocyte-predominant Hodgkin's disease (Hodgkin's paragranuloma), making the distinction between them extremely difficult. Immunohistochemistry provides a means of overcoming this difficulty. Immunostaining with UCHL1, L26, MB1, and 4KB5 was performed on five T-cell-rich B-cell lymphomas and 11 Hodgkin's paragranulomas (7/11 nodular, 4/11 diffuse). L26 stained the tumour cells not only of T-cell-rich B-cell lymphomas, but also of L+H Hodgkin's disease. In contrast, MB1 as well as 4KB5 identified all of the neoplastic cells in 3/5 T-cell-rich B-cell lymphomas, but did not react with the L+H cells in 8/11 Hodgkin's paragranulomas. Some overlap of staining patterns became apparent in the remaining cases, with 2/5 T-cell-rich B-cell lymphomas showing the MB1 + /4KB5 + phenotype in a tumour cell subset only. Similarly, in 3/11 Hodgkin's paragranulomas, some MB1/4KB5-positive L+H cells occurred in addition to MB1/4KB5-negative L+H cells. These cases, nevertheless, could be distinguished from one another by the numbers of MB1/4KB5-positive background lymphocytes, which were scanty or absent in T-cell-rich B-cell lymphomas and more numerous in Hodgkin's paragranulomas.

A 63-year-old woman presented with a history of increasing dysphagia of about two weeks duration. Laryngoscopy revealed a nonulcerated supraglottic epitheliomatous lesion that morphologically appeared well-differentiated and distinctly oncocytoid. Although the tumour lacked any criteria for malignancy such as cellular atypia, pleomorphism or necroses, it recurred twice after primary surgery and later gave rise to multiple painful skin metastases. The diagnosis of an oncocytoid differentiated neuroendocrine carcinoma of the larynx (laryngeal carcinoid) was made. Misinterpretation of laryngeal carcinoids is common, but can be avoided if one is familiar with this rare variant of laryngeal neoplasms

A morphometric and immunohistochemical study was performed on 354 bone marrow trephine biopsies derived from 126 patients with chronic myeloid leukaemia (CML) before and after allogeneic bone marrow transplantation (BMT). The purpose of this investigation was to evaluate the macrophage population, including several subsets and their dynamics in the posttransplant period. In addition to the total CD68+ resident (mature) macrophages the so-called activated fraction identified by its capacity to express alpha-D-galactosyl residues, the pseudo-Gaucher cells (PGCs) and the iron-laden histiocytic reticular cells were also considered. Following immuno- and lectin-histochemical staining morphometric analysis was carried out on sequential postgraft bone marrow specimens at standardized intervals. Compared to the normal bone marrow and calculated per haematopoiesis (cellularity) an overall decrease of about 40-50% in the quantity of CD68+ macrophages and the BSA-I+ subpopulation was detectable in the early posttransplant period (9-45 days after BMT). Noteworthy was the temporal recurrence of PGCs in the engrafted bone marrow, which was not associated with a clonally transformed cell population or leukaemic relapse. Reappearance of postgraft PGCs was most prominent in the first 2 months after BMT. This conspicuous feature was presumed to be functionally associated with a pronounced degradation of cell debris following pretransplant myelo-ablative therapy (scavenger macrophages). Evidence for an activation of the BSA-I+ macrophage subset was derived from the identical carbohydrate-binding capacity shown by the PGCs. In the regenerating haematopoiesis shortly after BMT a significant correlation between the number of BSA-I+ macrophages and erythroid precursor cells was determinable. This result implicates a close functional relationship between postgraft reconstitution of erythropoietic islets and centrally localized activated macrophages. In conclusion, findings emerging from this study included the reappearance of PCGs in the engrafted bone marrow independently of a leukaemic relapse and the significant association of the activated BSA-I+ macrophage subset with the recovery of erythropoiesis.

Following bone marrow transplantation (BMT) investigations on the recovery of the B and T lymphocyte populations have focused on the peripheral blood and only marginally regard the bone marrow. An immunohistochemical and morphometric study was performed on 352 trephine biopsies derived from 123 patients with chronic myelogenous leukemia (CML) at standardized endpoints before and after allogeneic BMT and compared to a control group. The purpose of this investigation was to quantify the B-CD20(+) and T-CD45RO(+) lymphocyte subsets and to determine possible relationships with the occurrence of acute and chronic GVHD. Moreover, we studied the dynamics of lymphocyte repopulation in the post-transplant period, correlations with the total peripheral lymphocyte count and differences associated with sibling vs alternate HLA-compatible (unmanipulated) marrow grafts. Morphometric analysis revealed a very fast regeneration of CD45RO(+) and CD20(+) marrow lymphocytes in the first 2 weeks following BMT. In less than 2 months, in most patients, the post-transplant quantity of lymphocytes was comparable to that of the normal bone marrow. This finding was opposed to the profound depression of the absolute lymphocyte count in the peripheral blood. No relevant relationships could be calculated between engraftment status and the lymphocyte repopulation in the bone marrow. On the other hand, significant correlations were calculable between the development of (chronic and acute) GVHD including severity with the number of CD45RO(+) lymphocytes. In non-related graft constellations a more frequent evolution of acute grade III + IV GVHD was detectable. This complication was accompanied by an increased quantity of CD45RO(+) lymphocytes in the marrow.

T-cell-rich B-cell lymphoma is a particular variant of large B-cell lymphomas with the morphological hallmark of a small number of large neoplastic B-cells scattered in between a dense background of reactive T-lymphocytes, while histiocytes may be admixed in variable numbers. In the typical case, the neoplastic population resembles large germinal center cells including cells similar to the L+H-variants of Reed-Sternberg cells. The immunophenotype of these tumour cells is L26 + Leu-M1-BerH2-. Apart from these unifying features, the individual cases constitute a broad spectrum of various growth patterns, so that a multiplicity of different relations to other types of malignant lymphomas are discussed in the literature. This occurs to such an extent that it may be doubted, that one deals with a distinct and separate lymphoma entity. Moreover, a close relationship exists between T-cell-rich B-cell lymphoma and lymphocyte predominant Hodgkin's disease, because there are striking similarities between the two, and, in addition, coexistence of T-cell-rich B-cell lymphoma with Hodgkin's paragranuloma has been reported. It, therefore, seems conceivable that T-cell-rich B-cell lymphoma represents a developmental stage of lymphocyte predominant Hodgkin's disease. Be that as it may: There is no doubt that, presently, the nosological position of T-cell-rich B-cell lymphoma is unsettled and still remains to be clarified.

To evaluate treatment-related changes of the reticulin stain-measured fibrosis in Ph(1+)-CML, a clinicopathological study was performed on sequential trephine biopsies of the bone marrow following either interferon (IFN) or busulfan (BU) monotherapy. Using the monoclonal antibody CD61 for the identification of megakaryopoiesis and Gomori's silver impregnation method, number of megakaryocytes and density of argyrophilic (reticulin and collagen) fibers were determined by morphometry. We studied specimens from 26 patients with IFN-alpha 2b (including nine patients with additional IFN gamma) therapy and from 23 patients who had received BU. In both groups, repeated bone marrow biopsies (total 125) revealed a significant increase in the fiber content, as well as in the number of megakaryocytes during treatment. To assess the dynamics of myelofibrosis more precisely, computation of differences in the degree of fiber density between the first and last examination was carried out. Regarding the considerable variations in the biopsy intervals, a so-called myelofibrosis progression index (MPI) was calculated. Following this rationale, we were able to demonstrate that, in comparison to the BU-group, speed of progression of bone marrow fibrosis was significantly increased in CML patients treated with IFN. Preliminary statistical analysis indicated a relationship between myelofibrosis on admission, which was always associated with increased growth of megakaryocytes, and the MPI with survival. Even when these parameters were regarded, prognosis was significantly more favorable in the IFN-treated patients. The failure of IFN and BU to inhibit the evolution of myelofibrosis may be related to several conversely acting pathomechanisms. Among others, the inability of both therapeutic agents to reduce the number of megakaryocytes more effectively should be taken into consideration.

A high content of mast cells (MC) is considered characteristic of neurofibromas but not of malignant schwannomas and neurilemmomas. We examined the extent and reliability of this finding by counting MC in 61 peripheral nerve sheath tumors and in 103 non-neurogenic soft tissue sarcomas. We furthermore investigated correlations between the amount of MC and various features of the tumors (e.g. grades of malignancy). Neurofibromas had very high mast cell counts. However, this result only applied to about 70% of these tumors. Malignant schwannomas, malignant fibrous histiocytomas and leiomyosarcomas had remarkably high median values of MC counts with a wide dispersion within the histological groups. Synovial sarcomas were the only group that contained MC in every case, though often in small numbers. In univariate analyses the number of MC was negatively correlated to grades of malignancy, cellularity and mitotic activity of the sarcomas and tended to correlate positively to the amount of myxoid and collagenous connective tissue and lymphocytic infiltrates. Multiple linear regression analysis revealed a significant correlation to the grade of malignancy and the amount of connective tissue.

Acute megakaryoblastic leukemia (M7) recently included in the French-American- British (FAB) classification for acute leukemias [1] is a rare hematological disease with poor prognosis. We present a case of acute megakaryoblastic leukemia with poor response to cytostatic treatment and a review of the therapeutic attempts in the literature.

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A case of adenocarcinoma of the colon is reported, whose metastases exhibited a syncytiotrophoblastic differentiation that gradually increased with advancing distance from the primary. Immunohistochemically, syncytiotrophoblastic giant cells with strong positivity for beta-HCG could be demonstrated within the metastatic tissue. Also, there was beta-HCG positivity in a fraction of the atypical epithelial elements of the primary. The case is discussed with respect to the differential diagnosis, and the cause of the syncytiotrophoblastic differentiation is attributed to the phenomenon of genetic instability. The case shows that by virtue of genetic instability totally new differentiations may be elaborated, so that the phenotypical appearance of a given tumor does not necessarily allow conclusions as to its tissue of origin. Thus, observations such as the one presented here clearly indicate that the histogenetic principle of tumor classifications by far does not apply to all cancers and that, therefore, its value and validity are considerably limited.

Two cases of occult seminoma were observed which showed left-sided lymph node metastases without clinically demonstrable testicular tumors. The peculiar histology of the lymph node metastases, namely PAS-positive tumor cells with clear cytoplasm together with a granulomatous stromal reaction, was highly suggestive of seminoma. Despite lack of testicular enlargement in both cases, a left-sided orchiectomy was done. Histologically, intratubular seminomas were found which confirmed the original diagnostic suspicions derived from the morphology of the lymph node metastases. It is concluded, therefore, that in the case of lymph node metastases with the above-mentioned structure, occult seminoma should always be taken into consideration. Because of the modern beneficial therapeutic implications, detection of occult seminoma is very important.

The malignant granular cell tumor ist a rare soft tissue neoplasia which is chiefly localized in the sceletal muscles. The uncertain histogenesis gave rise to different terms. The clinical course is often protracted but fatal. Diagnosis, differential diagnosis, and treatment are discussed in the light of a case report concerning a malignant granular cell tumor in a 28-year-old man.

Sparks-prothesis of 3 mm diameter were implanted into inbred rats and withdrawn after one to ten weeks. They were then fixed in Formalin and embedded in Paraffin and Metachrylat. After cutting (2.5 mu), the histological and histochemical parameters were examined. Cellular infiltration was studied after a certain period of time. The basic results of the genesis of that infiltration are given as follows: during the first two weeks following the s.c. implantation, derivates of monocytes (cells which play an imminent and important role during arterial replacement) were found. After the third and fourth week collagenous and elastic fibers could be seen, the connective tissue became stronger. The most favorable time for grafting can be evaluated by these experiments.

Question: By means of antimacrophage sera it is possible to demonstrate partially common antigenicity of monocytes with peritoneal macrophages, reticulum cells capable of phagocytosis, mast cells and Kupffer's cells. The authors continued these studies important for the explanation of cytogenetical interrelations using a special method (see below) on epitheloid and giant cells of rats. The experiments aimed for the detection of possible partially common antigenicity with the above mentioned monocytes/macrophages system.

By a combined application of toluidine blue stain and naphthol AS-D chloroacetate esterase reaction on normal bone marrow smears metachromic and promyelocytic granules were demonstrated on the same cell. Precursors of basophils contain NASDCl-esterase activity, the intensity of which depended on the maturity of the cells. All transitional cells were found between non metachromatic promyelocytes with strong NASDCl-esterase activity and completely enzymatically negative metachromatic segmented basophils. It is concluded that basophilic granulocytes like monocytes, neutrophils, and eosinophils derive from non specific promyelocytes. Evidence has been presented for the transition of promyelocytic granules to specific basophilic granules. Our results confirm the dualistic view of the cytogenesis of leukocytes

Necrotising splenic arteritis with numerous extensive anaemic splenic infarcts was found in four cases of Goodpasture's syndrome with the necrotising variant of rapidly progressive glomerulonephritis. The disease was rapidly fatal. Early scanning of splenic infarcts for diagnostic, prognostic and therapeutic purposes of the fulminating form of Goodpasture's syndrome is recommended.

The present paper deals with the term specificity. The term is analyzed with respect to its linguistic deduction, as well as its mode of application to morphological objects. We come to the conclusion that specificity is not an absolute but rather a relative term, since it is graduable. Its meaning largely depends on the conditions under which it is applied to morphological objects. When applied to morphological criteria, the term specificity is concerned in most cases with differential diagnosis; it rarely refers to diagnosis. In other words: Under certain circumstances, specific criteria may be absolute and, therefore, proving, but usually they have merely a relative meaning in the sense of indicating and delineating.

Publisher Summary This chapter discusses morphological recognition of immunoblastomas and centroblastomas. 93 large cell lymphomas were investigated, and the number and position of the nucleoli was assessed for 200 tumor cells from each case. Twenty-six cases could not be evaluated because of technical reasons such as inadequate fixation and mechanical distortion of the tissue. The remaining 67 cases were studied. Eighteen disclosed tumor cells with a very finely dispersed nuclear chromatin. These cells were devoid of clearly visible and identifiable marginal or central nucleoli. Therefore, only the diagnosis of large cell lymphomas could be made. The proposed cytological criteria by which large cell lymphomas are allegedly to be subdivided were simply not present in 18 of 67 cases (27%). However, 49 of the cases had cells with visible and identifiable nucleoli. So, it was possible to analyze 200 tumor cells from each case with respect to the numbers and positions of the nucleoli. The chapter highlights the results for the total of 9800 cells, of which 42% had a solitary nucleolus. But only in 25.4% of the cells was the nucleolus placed centrally and therefore, only these cells could clearly be classified as immunoblasts.

To understand and correctly interpret the morphology of tumors of the kidneys, and in the case of malignancy the morphology of their metastases, it is of importance to know that the definite kidney of man is a true metanephros. This means it is of mesodermal origin (Hamilton and Mossmann, 1972). On the other hand, there is no doubt of the epithelial character of the tubular cells. The mesodermal origin of epithelial cells makes it understandable that in renal neoplasms epithelial and nonepithelial, e.g., mesenchymal elements, may be found, for instance in the case of Wilms’ tumor, but also in the case of adenocarcinoma of the kidney.

Summary In chronic granulocytic leukaemia, hybridoid leucocytes can regularly be found. Light microscopically they contain a mixture of eosinophilic, basophilic and naphthol AS-D chloroacetate esterase-positive granules. The present study was done to clarify the ultrastructural composition of these cells. It could be clearly shown that in some leukaemic granulocytes primary and secondary eosinophilic as well as basophilic granules occur side by side. There were also basophils with additional tissue mast cell granules. Since normal mast cell granules as well as granules of normal eosinophilic promyelocytes are naphthol AS-D chloroacetate esterase-positive, it would appear possible that mastocytoid as well as primary eosinophilic granules within the leukaemic basophils are responsible for the atypical, granular naphthol AS-D chloroacetate esterase-positivity of these cells. The existence in chronic myeloid leukaemia both of mixed basophilic and eosinophilic granulated leucocytes and of mixed basophilic and mastocytoid granulated leucocytes may suggest a common myeloid precursor of eosinophils, basophils and tissue mast cells. In addition, the hybridoid granulocytes may be considered an expression of a neoplasiarelated lineage infidelity.

A case of myeloma showed histologically a monotonous pattern of relatively mature myeloma cells and an IgG-kappa paraproteinemia. In contrast to ordinary stains, the immunoperoxidase reactions for kappa and gamma gave inhomogeneous results. Light and heavy chains were formed by clusters of different subpopulations of myeloma cells. Serial sections showed that kappa-positive clusters were gamma-negative and vice versa. Thus, this morphologically homogeneous myeloma turned out to be immunohistochemically heterogeneous. We assume that two subpopulations have developed that produced separately kappa- and gamma-chains. Therefore, these chains might have been circulating separately and not as complete IgG-kappa molecules. Guided by our findings, the results of other investigators are discussed. Some considerations on the validity of immunohistochemical demonstration of tumor cell monoclonality are given implicating the diagnostic equivocation of immunohistochemistry for non-Hodgkin's lymphomas, especially for so-called “immunocytomas”.

A female patient presented with a lump in the right parotedeal region. A needle aspiration biopsy was performed which revealed numerous monomorphous atypical lymphoid cells with PAS-negative cytoplasmic inclusions. This raised the suspicion of malignancy. Therefore, the tumour was excised. Histologically, a nodular lymphoma of low grade of malignancy was diagnosed. The tumour contained many epithelioid histiocytes, sometimes with PAS-positive material. The case was difficult to classify after the Kiel-nomenclature, whilst classification on the basis of Rappaport's system was much easier.