Prognostic significance of polo-like kinase (PLK) expression in non-small cell lung cancer.

Our previous data indicate that the expression of the PLK gene which codes for a serine/threonine kinase is restricted to proliferating cells. In Northern blot experiments PLK mRNA expression was at the limit of detection in normal lung tissue but elevated in most samples of non-small cell lung cancer (NSCLC). A very low frequency of PLK transcripts was only found in bronchiolo-alveolar carcinomas. NSCLC patients whose tumors showed moderate PLK expression survived significantly longer (5 year survival rate=51.8%) than those with high levels of PLK transcripts (24.2%, P=0.001). No statistically significant correlation was found between PLK mRNA expression and age, sex, TNM status, histological type or degree of differentiation. Interestingly, the prognosis of patients in post-surgical stages I and II was correlated with PLK expression (5 year survival rates in stage I: 69.1% (moderate PLK) - 43.5% (high PLK), P=0.03 or in stage II: 51.9% (moderate PLK) - 9.9% (high PLK), P=0.006). These results suggest that PLK mRNA expression provides a new independent prognostic indicator for patients with NSCLC.

Littoral cell angioma. A novel splenic vascular lesion demonstrating histiocytic differentiation.

Seventeen cases of a novel type of vascular tumor of the spleen are described. The lesions, whose size ranges from minute foci to large nodules almost completely replacing the splenic tissue, are composed of anastomosing vascular channels resembling splenic sinus and have irregular lumina, often featuring papillary projections and cyst-like spaces; they are lined by tall endothelial cells that slough off into the vascular lumina and show hemophagocytosis. Atypical cells are absent and mitotic activity very low. In contrast to normal sinus endothelia, which express only FVIIIag, neoplastic cells express both endothelial (FVIII-AG, BMA 120) and histiocytic (KP1, lysozyme) antigens; occasionally S-100 protein is also present. The morphologic and immunohistochemical findings in this tumor reflect the dual differentiation potential of the reticuloendothelial cells lining the splenic sinus, justifying the term littoral cell angioma, and recognize a distinct entity that is different from other vascular lesions of the spleen, notably angiosarcoma. This distinction is all the more important because the clinical behaviour of this lesion is apparently benign.

Effect of aortic sclerosis on bone mineral measurements by dual-photon absorptiometry.

Measurements of the bone mineral content (BMC) of lumbar spine by dual-photon absorptiometry (DPA) are performed mainly in the anteroposterior (AP) projection. Due to superimposition of the abdominal aorta, the BMC measured for patients with aortic calcification usually is too high. To determine the influence of aortic calcifications, DPA scans were performed in the AP-projection on 100 dissected abdominal aortae with different degrees of atherosclerosis placed on a human lumbar spine cast in lucite. The measured values were compared with those obtained in the same projection without the aortae. The average increase of the BMC values relative to the mean for the vertebrae L2 to L4 for aortae with severe complicated lesions, i.e., those containing larger amounts of calcium, was 0.03 g/cm2, with a maximum deviation of 0.09 g/cm2. Aortae with fatty streaks or fibrous plaques did not cause significant increases of the BMC. The mean deviation for aortae with mild complicated lesions, i.e., those containing smaller amounts of calcium, was within the range of instrument precision.

Primary large cell lymphoma of the splenic sinuses: a variant of angiotropic B-cell lymphoma (neoplastic angioendotheliomatosis)?

A case of large cell lymphoma of B-cell lineage originating in the splenic sinuses is described. In addition to widening the spectrum of primary malignant lymphomas of the spleen, this case raises the possibility that variants of angiotropic large cell lymphomas may exist that do not involve blood vessels but do involve the spleen and lymph node sinuses.

Bone marrow findings after treatment with recombinant human interleukin-3.

In a phase I/II study, bone marrow biopsy specimens and aspirates of 20 patients with malignant tumors but normal bone marrow (n = 6), bone marrow failure resulting from chemotherapy (n = 4), myelodysplastic syndrome (n = 5), and aplastic anemia (n = 5) were evaluated before and after patients were treated with recombinant human interleukin-3 (rhIL-3). This cytokine proved to be an effective hematopoietic growth factor with only mild side effects. The rhIL-3 treatment led to increased overall bone marrow cellularity with trilinear stimulation of hematopoietic cells, except in most patients with aplastic anemia. In all patients, significant eosinophilia and, in some instances, bone marrow fibrosis developed. In addition to the increase in the number of circulating neutrophilic granulocytes, platelets, and reticulocytes, an increase of peripheral blood monocytes and lymphocytes was observed. The histologic and cytologic findings support the concept that rhIL-3 stimulates the proliferation and differentiation of pluripotent hematopoietic progenitor cells. It appears to be a safe and efficient therapeutic modality in patients with bone marrow failure. Additional clinical studies are needed to determine which patients will profit most from rhIL-3 treatment.

Identification of placental cytokine-producing cells in term and preterm labor.

OBJECTIVE: To determine if the production of proinflammatory cytokines by placentally derived macrophages changes with term and preterm labor and to examine if changes in antigen expression of these cytokines can be detected by immunohistologic methods. METHODS: Enzymatically dispersed placental cell suspensions of the trophoblastic villi, obtained from 16 women with spontaneous term delivery, 16 women with elective cesarean delivery without any labor, and 22 preterm delivering women with labor unresponsive to tocolysis, were fractionated by magnetic-associated-cell-sorting, on the basis of CD11b-antigen expression. Positively and negatively sorted cell fractions were cultured and concentrations of interleukin-6, interleukin-1beta, and tumor-necrosis-factor-alpha were measured in the culture supernatants. Immunohistologic staining was used for identification of cytokine-producing cells within placental tissues. RESULTS: Positively sorted cells obtained from term (median 2027 pg/mL, P = .037) and preterm (median 3628 pg/mL, P = .001) laboring women produced significantly elevated amounts of tumor-necrosis-factor-alpha compared with nonlaboring (median 1088 pg/mL) women at term. Negatively sorted cell fractions obtained from term (median interleukin-1beta 162 pg/mL, P = .031, median interleukin-6 3134 pg/mL, P = .004) and preterm (median interleukin-1beta 934 pg/mL, P = .003, median interleukin-6 5695 pg/mL, P = .001) laboring women produced significantly elevated amounts of interleukin-1beta and interleukin-6 compared with nonlaboring (median interleukin-1beta 29 pg/mL, median interleukin-6 135 pg/mL) women at term. Immunohistologic staining revealed that tumor-necrosis-factor-alpha activity was localized in isolated stromal cells, whereas interleukin-1beta and interleukin-6 were predominantly found in endothelial cells within placental villi. CONCLUSION: The source of labor-associated release of tumor-necrosis-factor-alpha from placental tissues are macrophages, whereas interleukin-1beta and interleukin-6 are released from placental endothelial cells.

Peripheral T-cell lymphomas respond well to vincristine, adriamycin, cyclophosphamide, prednisone and etoposide (VACPE) and have a similar outcome as high-grade B-cell lymphomas.

Peripheral T-cell lymphomas (PTCL) represent a heterogeneous group of T-cell malignancies including subentities with favourable (large cell anaplastic) or unfavourable (pleomorphic) prognosis. The clinical outcome of PTCL has been controversially discussed, but a worse prognosis than high-grade B-cell Non-Hodgkin's lymphomas (NHL) has been postulated by most authors. In this report we summarize the results of a prospective comparative study investigating the therapy outcome of 27 patients (pts) with PTCL and 55 pts. with high grade B-cell NHL and give an overview of therapy studies in PTCL. The histological subtypes were 14 pleomorphic, 8 large-cell anaplastic (Ki-1+), 2 angioimmunoblastic (AILD) and 3 other PTCL. In three patients the PTCL was associated with non-tropical sprue (11%). Nineteen patients presented with an advanced stage of disease (stage III and IV, 70%), 17 (63%) pts. had B-symptoms. The patients were treated with vincristine 2 mg d1, adriamycin 25 mg/m2 d1-3, cyclophosphamide 800 mg/m2 d1, prednisone 60 mg/m2 d1-7 and etoposide 120 mg/m2 d1-3 (VACPE). In 77% of pts. with PTCL and 84% of patients with high-grade B-cell NHL a complete remission (CR) was achieved. 75% of the complete responders with PTCL and 70% with B-NHL are still in ongoing CR. The subgroup of large-cell anaplastic attained a CR in 88%. The median observation time is 44 months (1(+)-77+). The probability of 1-, 3- and 5-year overall and disease-free survival for the T-cell group were 76%, 54%, 48% and 76%, 62%, 62%, respectively according to Kaplan-Meier. There was no significant difference regarding the remission rate, the overall-, event-free or disease-free survival compared to high-grade B-cell lymphomas. In conclusion, the VACPE regimen is an effective and feasible regimen in the management of PTCL achieving complete remissions in a large proportion of patients.

Splenic metastasis in an ileal carcinoid tumor.

The case of an ileal carcinoid tumor metastatic to the spleen causing splenomegaly and symptoms that necessitated splenectomy is described. Morphological analysis reveals that viable tumor cells are present along the whole pathway of splenic circulation and, accordingly, may lead to metastatic deposits at different sites within the spleen.

The spleen in acquired immunodeficiency syndrome (AIDS).

72 spleens from AIDS patients were studied by histologic and immunohistochemical methods. Apart from the opportunistic infections and malignancies typically occurring in AIDS patients (malignant lymphomas, Kaposi's sarcoma, mycobacterial infections, cryptococcosis, cytomegalovirus infections) morphological changes probably directly attributable to HIV infection were noted that progressed from follicular hyperplasia with nearly normal periarteriolar lymphoid sheaths to atrophy of follicles and severe cellular depletion of T cell areas with concomitant plasmacytosis and occurrence of small giant cells. At the same time the number of Protein S-100 positive cells markedly increased; in some cases almost all pulp cord macrophages expressed the antigen. In addition, the sinus endothelia strongly expressed protease inhibitors.

Histogenesis of Kaposi's sarcoma associated with AIDS: a histologic, immunohistochemical and enzyme histochemical study.

Twenty-one cases (25 biopsies including 9 frozen biopsies) of Kaposi's sarcoma associated with the acquired immune deficiency syndrome (AIDS) were examined immunohistochemically, lectin-histochemically, and enzyme histochemically to ascertain the histogenesis of the lesion. The Kaposi's sarcomas were histologically subtyped according to a modified Schmid's classification (granulation tissue-like-, angiosarcoma-like- and spindle cell type). In almost all lesions, many atypical vasoforming cells and at least some spindle cells without definite evidence of vasoformation by conventional microscopy were positive for factor VIII-related antigen, BMA 120 (a new monoclonal antibody to an endothelial cell-specific antigen), Ulex europaeus I (UEA-I), alkaline phosphatase and ATPase. Linear reaction products for BMA 120 and UEA-I, suggesting the luminal surface of immature vascular channels, were sometimes recognized in the positive spindle cells. Electron micrographs confirmed endothelial characteristics, such as irregular and fragmented but distinct basal lamina and numerous pinocytotic vesicles, in both the UEA-I- and ATPase-positive spindle cells. Among spindle cells negative for the endothelial markers, there were many macrophages as a stromal reaction to tumor tissue, identified by monoclonal antibodies to macrophages (KiM 6, 7, 8 and EBM 11), acid phosphatase and alpha-naphthyl acetate esterase. The results of the immuno- and enzyme histochemical investigations did not correlate with the different histologic types of Kaposi's sarcoma. However, our results strongly suggest that tumor cells of Kaposi's sarcoma are derived from vascular endothelial cells rather than lymphatic endothelium.

Disseminated visceral leishmaniasis (kala azar) in acquired immunodeficiency syndrome (AIDS).

The case of an AIDS patient with visceral leishmaniasis diagnosed by bone marrow and liver biopsy is reported. Despite the infrequent association between HIV infection and leishmaniasis pathologists and clinicians alike should be alerted to the possibility of leishmaniasis occurring as yet another opportunistic infection in the setting of HIV-induced immunodeficiency.

Urokinase plasminogen activator induces angiogenesis and tumor vessel invasion in breast cancer.

Urokinase plasminogen activator (uPA) is a proteolytic enzyme implicated in cancer invasion and tumor progression. Urokinase PA and its inhibitor (PAI-1) appear to be new and independent prognostic markers in breast cancer. To investigate how uPA- and PAI-1-levels correlate with angiogenesis and tumor vessel invasion, we counted microvessels and their tumor invasion and determined the uPA- and PAI-1 levels in 42 primary invasive breast carcinomas. 20 Patients had no lymph node metastasis at the time of surgery, while 22 patients had positive nodes. Using light microscopy, we highlighted the vessels by staining their endothelial cells immunocytochemically for CD31 and Factor VIII. After gaining tumor tissue extracts, we determined the uPA- and PAI-1-levels by ELISA. A positive correlation between microvessel density, angioinvasion and uPA- and PAI-1-levels was found. We speculate that high uPA levels may induce tumor neovascularisation, angioinvasion and may cause tumor progression and metastasis. The degradation of the vessel wall by uPA causes a leak. This wall defect may, on the one hand, be the stimulus for endothelial cell proliferation and formation of new blood vessels and, on the other hand, it may be the place of tumor cell entry.

Immuno- and enzymehistochemical characterization of 'plasmacytoid T-cells' in formalin-fixed paraffin-embedded tissue of reactive lymph nodes.

In eight (10.4%) of 77 randomly selected cases of lymphadenitis, foci of 'plasmacytoid T-cells' (PTCs) were found. We characterized the PTCs in formalin-fixed paraffin-embedded tissue by immuno- and enzymehistochemical techniques. In each case the PTCs formed nodular and patchy infiltration patterns around the high endothelial venules in the paracortex of the lymph nodes. The PTCs in half of the cases were present near the intermediary peritrabecular sinuses. The PTCs were found to be positive for CD 45 (LCA), CD 45 R (4 KB 5) and CD 43 (MT 1), but not for CD 45 RO (UCHL 1), L 26 (CD 20), myeloperoxidase, naphthol AS-D chloroacetate esterase, Mac 387, alpha-1-antichymotrypsin or S 100 protein and anti-dendritic reticulum cell antibody. Some lysozyme+ PTCs were mingled in and around the foci in each case. In five cases the PTCs showed weaker positive reactions for CD 68 (EBM 11) and HLA-DR than for T-zone macrophages and sinus histiocytes. Related to the degree of desintegration of the nodular PTC-foci increasing numbers of T- and B-lymphocytes, T-zone macrophages and interdigitating reticulum cells with HLA-DR were detected in the foci. In two cases, where there was no HLA-DR expression by PTCs, small amounts of HLA-DR in secondary lymph follicles and a decrease of HLA-DR+ T-zone macrophages and interdigitating reticulum cells were observed. Our results suggest that there is a high degree of interaction between PTCs and other cellular components in the lymph node. The PTC may represent a special type of resident macrophage with secretory capacities.

In vitro cytokine secretion and maturation phenotype of lymphoma-associated splenic macrophages.

In this study, cytokine secretion capacity and maturation phenotype of human lymphoma-associated splenic macrophages (LASM) were evaluated in a long-term culture. Sixteen spleens from malignant lymphoma patients and five control spleens were investigated. Splenic macrophages (SM) were isolated by teflon adherence and cultured for 6-48 days. Secretion of IL-1 alpha, IL-6 and TNF alpha was measured by ELISA following maximal stimulation with LPS and IFN-gamma, and cytokine mRNA expression was detected by in situ hybridization. Immunohistochemical expression of maturation-associated antigens was evaluated semiquantitatively. Cytokine secretion capacity was significantly altered in LASM which exhibited reduced TNF alpha and IL-6, but elevated IL-1 alpha secretion when compared to control SM. Alterations of cytokine secretion capacity were associated with a modification of LASM maturation phenotype, showing impaired expression of early and chronic/late inflammatory markers. These findings obtained from a long-term culture model suggest that malignant lymphomas induce lasting modifications of cytokine secretion and maturation patterns in LASM.

Polo-like kinase: a novel marker of proliferation: correlation with estrogen-receptor expression in human breast cancer.

Previous data have shown that the mRNA-expression of the serin/threonine-kinase polo-like kinase (PLK) is closely correlated with the survival of patients suffering from a subset of malignant tumors. PLK-mRNA and protein-expression are restricted to cells in the cell cycle. PLK-mRNA-transcripts are highly abundant in proliferating cells; no gene expression is found in G0-phase cells. Here we investigated the mRNA- and protein-expression of PLK- and estrogen-receptor (ER) in human breast-carcinoma by northern-blotting, RT-PCR and immunohistochemistry. The expression of MIB-I was determined on serial sections. Analysis of the immunohistochemical data revealed a close correlation between the ER and PLK-expression (r = 0.677; p = 0.001, n = 30). No relationship between the mRNA-expression of ER and PLK was found. Furthermore, no correlation for the protein expression of PLK and MIB-I exists. The influence of estrogen (ES) is known to have proliferative potential. The expression of ER correlates with the ES-plasma-level. In addition, the hormone cycle of premenopausal women undergoes rapid vacillations with varying effects on the proliferating tumor cells, e.g., growth induction. Our results therefore show that ER-expression is not only of therapeutic value for the clinician, but it may also be a tool for determining the tumor proliferation index more precisely by integrating the hormone-mediated proliferation stimulus.

Familial erythrophagocytic lymphohistiocytosis (Farquhar's disease): involvement of the central nervous system.

The clinical course and the postmortal pathological findings in a female newborn showing parental consanguinity are presented. One week afterbirth, the infant developed fever, hepatosplenomegaly and polyserositis. Rapidly progressing immunodeficiency due to pancytopenia led to pneumonia and untreatable respiratory distress with fatal outcome after 2 weeks. Autopsy findings revealed multisystem lymphohistiocytic infiltration with marked erythrophagocytosis. Neuropathological findings included lymphohistiocytic leptomeningitis, perivascular cuffing by lymphohistiocytic infiltrations in the cerebral white matter, predominantly in subependymal location, and multifocal lymphohistiocytic infiltrations of the cerebral grey matter and the cervical spinal cord. Erythrophagocytosis was the histopathological hallmark at all sites. Regarding the fatal clinical course, the medical history of parental consanguinity and the histopathological features, postmortem diagnosis was familial erythrophagocytic lymphohistiocytosis (FEL, Farquhar's disease). The present case is discussed with focus on CNS involvement in FEL by reviewing the relevant literature.

Reversal of acute vascular rejection by plasma exchange.

The vast majority of grafts with acute vascular rejection (AVR) undergo irreversible loss of function, since this type of rejection usually does not respond to increased levels of immunosuppression. Since 1980 we have treated with plasmafiltration (PF) 14 steroid resistant episodes of AVR in 10 patients. In the treatment of each episode 6 X 5 L plasma were filtered and simultaneously replaced with 3.5% albumin-Ringer-lactate solution. After each PF, 20 g immunoglobulins (Intraglobin) were replaced i.v. In all cases following PF renal function improved. One graft was eventually lost as a result of chronic irreversible vascular rejection; all others are still functioning. Eight of the 10 patients have been observed now for at least 6 months since beginning PF treatment and 5 of these 8 have been followed for more than 12 months. None of them shows signs of rejection. PF with replacement of immunoglobulins after treatment appears to be a simple and safe procedure, which in our hands proved to be quite effective in reversing AVR.

Chronic recurrent appendicitis. A clinicopathologic study of 47 cases.

As part of a retrospective study 47 appendices from patients with a clinical and morphologic diagnosis of chronic recurrent appendicitis were studied by morphologic and immunohistochemical methods. Appendices with chronic recurrent appendicitis histologically show chronic active inflammation with increased numbers of immunocompetent cells, mostly T lymphocytes and subsequent scarring. In addition, there is an increase in the number of neural cells. Apparently the secretion of neurotropic inflammatory mediators, especially by activated and degranulating eosinophils, leads to a stimulation of nociceptive receptors and thus to the clinical symptoms. The diagnosis of recurrent chronic appendicitis is thus warranted in these cases not only clinically but also on the basis of histologic and immunohistochemical findings.