Transformation potential of bone marrow stromal cells into undifferentiated high-grade pleomorphic sarcoma

Purpose

Bone marrow adherent cells contain conventional bone marrow stromal cells and mesenchymal stem cells and these cells constitute the hematopoietic microenvironment. Mesenchymal stem cells have the capacity to give rise to multiple mesenchymal lineage cells and even ectodermal lineage cells. In the present study, we investigated what types of tumor cells are inducible from BM adherent cells by chemical carcinogens.

Methods

Bone marrow cells from neonatal C3H/HeN mice were collected within 24 h after birth and then cultured. Four days later, bone marrow adherent cells were obtained and the cells were treated with 3-methylcholanthrene.

Results

By this treatment, some transformed clones consisting of large spindle cells were obtained. The transformed cells were highly positive for CD44 and were positive for Sca-1, CD49d and CD106, whereas the cells were negative for hematolymphoid markers. The cell clones had the ability to support hematopoiesis in vitro. These results indicate that the transformed cell lines have the characteristics of BM stromal cells/mesenchymal stem cells. Moreover, during culture of the transformed cells, spontaneous bone nodule formation was observed. When the transformed cells were inoculated into immunodeficient mice subcutaneously, the neoplasms grew in the subcutaneous tissue of the mice. Microscopically and ultrastructurally, the neoplasms showed the typical morphology of undifferentiated high-grade pleomorphic sarcoma (UHGPS). Bone-related genes have been found to be expressed in both transformed cells and UHGPSs.

Conclusion

The present study suggests that UHGPSs are derived from BM stromal cells, probably mesenchymal stem cells.     

Low ADAMTS-13 activity during hemorrhagic events with disseminated intravascular coagulation

Disseminated intravascular coagulation (DIC) is a life-threatening complication, and its control is essential for therapeutic success. Recombinant human soluble thrombomodulin alfa (rTM) is a novel therapeutic agent for DIC. The efficacy of rTM in the treatment of DIC is reportedly superior to that of conventional anti-DIC treatments, such as unfractionated heparin or low molecular weight heparin, but hemorrhagic events occasionally interfere with the therapeutic benefits of rTM. We assessed the clinical features of 20 consecutive patients who were given rTM for DIC associated with various hematologic disorders. Eight patients achieved remission of both primary disease and DIC, eight died due to progression of the primary disease, and four died of various hemorrhagic complications. Assessment of 16 biomarkers for coagulation showed that the four patients who died of hemorrhagic complications despite remission of their primary disease showed lower ADAMTS-13 (a disintegrin and metalloproteinase with a thrombospondin Type 1 motif, member 13) plasma activity than other patients (P = 0.016). The optimal cut-off level of ADAMTS-13 for predicting risk of hemorrhagic complications was 42 % (P = 0.007). Plasma ADAMTS-13 activity determined at diagnosis of DIC may help predict the risk of hemorrhagic events during and/or following DIC treatment with hematologic disorders.     

The extract from Nandina domestica THUNBERG inhibits histamine- and serotonin-induced contraction in isolated guinea pig trachea

Although the fruit of Nandina domestica THUNBERG (ND) has been used to treat respiratory disorders such as coughing and breathing difficulty in Japan for many years, very little is known about mechanisms underlying its action. In the present study, we investigated effects of the crude extract from ND (NDE) and one of its constituents, nantenine, on contractile responses in isolated guinea pig tracheal ring preparations. In normal experimental condition, guinea pig trachea remained tonically contracted during the resting state, and addition of NDE (1 mg/ml) caused a relaxation of tracheal smooth muscles, but had little effect on the responsiveness of trachea to acetylcholine. The basal, tonic contraction was abolished by the presence of atropine and indomethacin. In this condition, NDE at 0.1-1 mg/ml inhibited histamine-induced contraction in both competitive and non-competitive manners. NDE at 0.01-1 mg/ml inhibited serotonin-induced contraction in a competitive manner. Nantenine (2-20 microM) did not affect histamine-induced contraction, and slightly inhibited serotonin-induced contraction. These results suggest that NDE has inhibitory effects on tracheal smooth muscle contraction, and nantenine cannot account solely for this effect of NDE.     

Association between passive smoking and salivary markers related to periodontitis

OBJECTIVES: The mechanism of passive smoking in terms of development of periodontitis has not been investigated. This study examined the effect of passive smoking on salivary markers related to periodontitis. METHODS: Periodontal status was evaluated on the basis of probing pocket depth and clinical attachment level in 273 workers. Salivary marker levels were determined by enzyme assay including enzyme-linked immunosorbent assay. Six periodontal pathogens in saliva were assessed using real-time PCR methodology. Non-, passive and active smokers were defined as subjects exhibiting salivary cotinine levels of 0 (53 subjects), 1-7 (118) and > or = 8 ng/ml (102). RESULTS: Levels of salivary markers, including IL-1beta, lactoferrin, albumin and aspartate aminotransferase (AST), were elevated significantly in passive smokers relative to non-smokers. Additionally, these marker levels, with the exception of IL-1beta, decreased significantly in active smokers in comparison with passive smokers. However, no meaningful differences in percentages of periodontal pathogens were observed between non- and passive smokers. Multiple linear regression analyses were performed for each marker utilizing age, gender, cotinine level and periodontal status as independent variables. IL-1beta, albumin and AST were independently associated with cotinine level. CONCLUSION: Passive smoke exposure leads to elevation of IL-1beta, albumin and AST levels in saliva.     

Statins,inhibitors of 3‐hydroxy‐3‐methylglutaryl‐coenzyme A reductase,function as inhibitors of cellular and molecular components involved in type I interferon production

Objective

Statins, which are used as cholesterol‐lowering agents, have pleiotropic immunomodulatory properties. Although beneficial effects of statins have been reported in autoimmune diseases, the mechanisms of these immunomodulatory effects are still poorly understood. Type I interferons (IFNs) and plasmacytoid dendritic cells (PDCs) represent key molecular and cellular pathogenic components in autoimmune diseases such as systemic lupus erythematosus (SLE). Therefore, PDCs may be a specific target of statins in therapeutic strategies against SLE. This study was undertaken to investigate the immunomodulatory mechanisms of statins that target the IFN response in PDCs.

Methods

We isolated human blood PDCs by flow cytometry and examined the effects of simvastatin and pitavastatin on PDC activation, IFNα production, and intracellular signaling.

Results

Statins inhibited IFNα production profoundly and tumor necrosis factor α production modestly in human PDCs in response to Toll‐like receptor ligands. The inhibitory effect on IFNα production was reversed by geranylgeranyl pyrophosphate and was mimicked by either geranylgeranyl transferase inhibitor or Rho kinase inhibitor, suggesting that statins exert their inhibitory actions through geranylgeranylated Rho inactivation. Statins inhibited the expression of phosphorylated p38 MAPK and Akt, and the inhibitory effect on the IFN response was through the prevention of nuclear translocation of IFN regulatory factor 7. In addition, statins had an inhibitory effect on both IFNα production by PDCs from SLE patients and SLE serum–induced IFNα production.

Conclusion

Our findings suggest a specific role of statins in controlling type I IFN production and a therapeutic potential in IFN‐related autoimmune diseases such as SLE.

     

Worsened long-term outcomes and postoperative complications in octogenarians with lung cancer following mediastinal lymph-node dissection

We evaluated the effects of mediastinal lymph-node dissection on outcomes in octogenarians with primary lung cancer. Outcomes and postoperative complications were retrospectively investigated in 48 octogenarians with anatomically resected lung cancer, of whom 23 underwent a mediastinal lymph-node dissection (ND2 group) and 25 a limited lymphadenectomy (ND0-1 group). Forty-three patients underwent a lobectomy, two a pneumonectomy, and three a segmentectomy. The five-year survival rate for all was 35%, while that for those in pathological stage I was 43.3% and for those in stage II+III was 21.2%. As for lymph node dissection, the five-year survival rate for the ND0-1 group (54.3%) was superior to that for the ND2 group (21.7%) (P=0.022). For patients in pathological stage I, those rated ND0-1 had a better five-year survival than those rated ND2 (61.9% vs. 28.6%) (P=0.041). In addition, mediastinal lymph-node dissection increased the incidence of postoperative cardiac complications (P=0.004). Our results indicate that major pulmonary resection with mediastinal lymph-node dissection is associated with a higher rate of mortality in octogenarians with lung cancer.     

Preferential expansion of human umbilical cord blood-derived CD34-positive cells on major histocompatibility complex-matched amnion-derived mesenchymal stem cells

Background

We previously found in a murine hematopoietic system that hematopoietic stem cells show high differentiation and proliferation capacity on bone marrow-derived mesenchymal stem cells/stromal cells (microenvironment) with “self” major histocompatibility complex (MHC).

Design and Methods

We examined whether amnion-derived adherent cells have the characteristics of mesenchymal stem cells, and whether these adherent cells can support the proliferation of umbilical cord blood-derived lineage-negative and CD34-positive cells (Lin^–CD34⁺ cells) obtained from the same fetus to a greater extent than those derived from other fetuses.

Results

Culture-expanded amnion-derived adherent cells expressed mesenchymal stem cell markers and HLA-ABC molecules and could differentiate into osteoblasts, adipocytes and chondrocyte-like cells, indicating that the cells have the characteristics of mesenchymal stem cells. The Lin^–CD34⁺ cells purified from the frozen umbilical cord blood were strongly positive for HLA-ABC, and contained a large number of hematopoietic stem cells. When the Lin^–CD34⁺ cells were cultured on the autologous (MHC-matched) or MHC-mismatched amnion-derived adherent cells in short-term assays (hematopoietic stem cell-proliferation) and long-term culture-initiating cell assays, greater expansion of the Lin^–CD34⁺ cells was observed in the MHC-matched combination than in MHC-mismatched combinations. The concentration of granulocyte-macrophage colony-stimulating factor in the culture supernatants of the long-term culture-initiating cell assays was significantly higher in the MHC-matched combination than in MHC-mismatched combinations.

Conclusions

It is likely that a MHC restriction exists between hematopoietic stem cells and mesenchymal stem cells/stromal cells in the human hematopoietic system and that granulocute-macropage colony-stimulating factor contributes to some extent to the preferential hematopoiesis-supporting ability of the MHC-matched amnion-derived adherent cells.     

Leiomyomatosis peritonealis disseminata coexisting with endometriosis within the same lesions: a case report with review of the literature

Leiomyomatosis peritonealis disseminata (LPD) is an extremely rare condition, which is characterized by the presence of multiple peritoneal and subperitoneal nodules composed of bland smooth muscle cells. Albeit extremely rare, coexistence of endometriosis within LPD lesions has also reported. Herein, we report the seventh documented case of LPD coexisting with endometriosis within the same lesions and review the pathogenesis of this lesion. A 42-year-old Japanese female presented with an abdominal tumor. Computed tomography revealed a tumorous lesion in the right ovary and multiple small nodules in the abdominal cavity. Under a clinical diagnosis of ovarian cancer with peritoneal dissemination, resection of these lesions was performed. Histopathological study of the disseminated peritoneal nodules revealed proliferation of interlacing bundles of spindle cells with eosinophilic cytoplasm and bland cigar-shaped nuclei. Mitotic figures were hardly seen. The peritoneal nodules of the rectum had cystic cavities within the spindle cell bundles, and endometrial glands and stroma were present around the cystic cavities and spindle cells. The resected tissues of the ovary and cecum showed the same histopathological features. Accordingly, a diagnosis of LPD with endometriosis within the same lesions was made. A possible origin of LPD is thought to be the submesothelial multipotential stem cells, also referred to as the secondary müllerian system. The presence of endometrial tissues within LPD lesions, as seen in the present case, also support this hypothesis because endometrial tissues are also derived from the müllerian system.