Genomic alterations in primary cutaneous melanomas detected by metaphase comparative genomic hybridization with laser capture or manual microdissection: 6p gains may predict poor outcome

To clarify the correlation of genomic alterations with clinical and histological features, we performed metaphase comparative genomic hybridization analysis on 20 primary cutaneous melanomas, which were obtained by laser capture or manual microdissection, and 16 melanoma cell lines. There were no differences in the average number of aberrations between acral melanomas (AM) and non-AM, although gains of 5q and 11q13 were more frequent (P=0.05) and 10q loss was less frequent (P=0.01) in AM than in non-AM. Although tumor thickness is considered a measurable estimate of clinical expression, there were no differences in the average number of aberrations among 4 groups, classified by thickness of the tumor. While the majority of aberrations were equally distributed among the 4 groups, 6p gains were found only in the thickest tumors. Patients with 6p or 1q gains had a lower overall survival rate than those without them (P=0.0002 or P=0.013). While gains of 1q, 2q, 3p, 3q, 7q, 20p, and 20q were more frequent in the cell lines than in the primary tumors (P<0.01), losses of 6q, 9p, 10p, and 10q were equally found in both cell lines and primary tumors. The present study showed that chromosomal aberrations had already occurred in the thinner tumors, and that 6p and 1q gains may be a prognostic factor.     

Therapeutic effect of topical administration of SN50, an inhibitor of nuclear factor-kappaB, in treatment of corneal alkali burns in mice

We evaluated the therapeutic efficacy of topical administration of SN50, an inhibitor of nuclear factor-kappaB, in a corneal alkali burn model in mice. An alkali burn was produced with 1 N NaOH in the cornea of C57BL/6 mice under general anesthesia. SN50 (10 microg/microl) or vehicle was topically administered daily for up to 12 days. The eyes were processed for histological or immunohistochemical examination after bromodeoxyuridine labeling or for semi-quantification of cytokine mRNA. Topical SN50 suppressed nuclear factor-kappaB activation in local cells and reduced the incidence of epithelial defects/ulceration in healing corneas. Myofibroblast generation, macrophage invasion, activity of matrix metalloproteinases, basement membrane destruction, and expression of cytokines were all decreased in treated corneas compared with controls. To elucidate the role of tumor necrosis factor (TNF)-alpha in epithelial cell proliferation, we performed organ culture of mouse eyes with TNF-alpha, SN50, or an inhibitor of c-Jun N-terminal kinase (JNK) and examined cell proliferation in healing corneal epithelium in TNF-alpha-/- mice treated with SN50. An acceleration of epithelial cell proliferation by SN50 treatment was found to depend on TNF-alpha/JNK signaling. In conclusion, topical application of SN50 is effective in treating corneal alkali burns in mice.     

Biological activities of Bacteroides forsythus lipoproteins and their possible pathological roles in periodontal disease

Bacteroides forsythus is a gram-negative, anaerobic, fusiform bacterium and is considered to be an etiological agent in periodontal disease. A lipoprotein fraction prepared from B. forsythus cells by Triton X-114 phase separation (BfLP) activated human gingival fibroblasts and a human monocytic cell line, THP-1, to induce interleukin-6 production and tumor necrosis factor alpha production. BfLP was found to be capable of inducing nuclear factor-kappaB translocation in human gingival fibroblasts and THP-1 cells. By using Chinese hamster ovary K1 cells transfected with Toll-like receptor genes together with a nuclear factor-kappaB-dependent CD25 reporter plasmid, it was found that signaling by BfLP was mediated by Toll-like receptor 2 but not by CD14 or Toll-like receptor 4. BfLP induced apoptotic cell death in human gingival fibroblasts, KB cells (an oral epithelial cell line), HL-60 cells (a human myeloid leukemia cell line), and THP-1 cells but not in MOLT4 cells (a T-cell leukemia cell line). Caspase-8, an initiator caspase in apoptosis, was found to be activated in these cells in response to BfLP stimulation. Thus, this study suggested that BfLP plays some etiological roles in oral infections, especially periodontal disease, by induction of cell activation or apoptosis.     

ISCHEMIC ENTEROCOLITIS WITHOUT ARTERIO–OCCLUSIVE LESION

Four cases of Ischemic enterocolitis without arterio–occlusive lesion were described. Three cases were associated with sigmoid colon carcinomas. Ischemic lesions developed anal to the carcinomas in two cases, and oral to sigmoidostomy to relieve intestinal obstruction by carcinoma in one case. One other case was associated with inguinal hernia. Grossly, ischemic lesions involved relatively short intestinal segments, and the ischemic colonic lesions were not related to teniae coli. Extensive veno–occlusive lesions were discovered in a case of ischemic stricture of the ileum, which had been incarcerated in the right inguinal hernia. Reversible mechanical occlusion of the intestinal vessels caused by transient or recurrent intestinal strangulation is the most probable cause of these ischemic lesions., ACTA PATHOL. JPN. 33: 249–256, 1983.     

Nondestructive evaluation of blood flow in a dialyzer using X-ray computed tomography

It is very important to observe the concentrations and flow patterns of blood through a dialyzer to evaluate its function and to obtain the most appropriate design. We established a visualization method for the blood flow pattern in a dialyzer using X-ray computed tomography, and investigated the so-called internal filtration phenomenon. The results obtained were as follows: (1) The influence of 5% BaSO₄, which was added to the blood as a contrast medium, on the filtration rate of the dialyzer was minimal. (2) The relationship between the concentration of BaSO₄ and the Hounsfield unit value was expressed by linear regression. (3) Hounsfield unit values increased massively just after blood entered the dialyzer and peak values increased with dialysate perfusion under the following conditions: the dialyzer (BS-1.6UL, polysulfone hollow fibers) was used, and bovine blood with 5% BaSO₄ added was used at a blood flow rate of 200ml/min. The dialysate flow rate was 500ml/min and the slice thickness of X-ray computed tomography was 1–10mm. (4) It was observed that blood flowed slightly faster in the center than the peripheral portion of the dialyzer, when the flow pattern was followed after pulse injection of blood containing 20% BaSO₄ into the dialyzer. It was concluded that this method could possibly be utilized not only qualitatively but also quantitatively for observation of the real state of blood flow and in designing dialyzers.     

Depression of the generation of cell-mediated cytotoxicity by suppressor cells after surgery

The effects of surgical operation on the generation of cell-mediated cytotoxicity in mixed cell cultures were studied in patients with various carcinomas or benign lesions. Peripheral blood mononuclear cells from patients were cultured with B lymphoblastoid cell line Raji in mixed culture, and the induced cytotoxicity was measured by 51Cr release assay. In 15 patients with various carcinomas, the capacity of cells to generate cytotoxic cells was significantly depressed 1, 3 and 6 days after surgery, as compared to that before surgery. It returned to the pre-operative level by the 8th post-operative day. In eight patients with benign lesions, significant decrease in cytotoxic cell activity was observed 3 and 6 days after operation. At the 8th day, however, there was a significant increase in the generated cytotoxicity. The depressed generation of cytotoxic cells 3 days after surgery could be abrogated by removal of adherent cells from the responding cell population. This effect could be partially reconstituted by addition of separated, autologous adherent cells back to the responding non-adherent cell culture. These results demonstrate that suppressor cells, presumably monocytes, may be responsible for the depressed generation of cytotoxic cells after surgery.     

Social isolation stress augments angiogenesis induced by colon 26-L5 carcinoma cells in mice

We have previously shown that tumor necrosis factor-α (TNF-α), which is an important angiogenesis-related factor, was over-secreted in male BALB/c mice under social isolation stress as compared with the control, and closely associated with a remarkable elevation of tumor invasion and metastasis of colon 26-L5 carcinoma cells. In the present study, we explored the effect of isolation stress on the angiogenesis caused by colon 26-L5 carcinoma cells in vivo and in vitro. Social isolation lead to the enhancement of tumor growth after intrahepatic implantation with a fragment of colon 26-L5 tumor. Angiogenic response (number of vessels oriented towards tumor mass) and tumor growth (size) were significantly increased in the socially isolated mouse relative to that in the group-housed mice. Furthermore, higher protein level of hepatic TNF-α was found in the stressed mice than that in the control. Expression of mRNA for vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) were also elevated in the tumor regions and liver tissues of the stressed mice in comparison with that in group-housed mice. On the other hand, hepatic sinusoidal endothelial (HSE) cells treated with TNF-α exhibited a marked promotion of the migration, invasion, expression of mRNA for matrix metalloproteinase (MMP)-9, and tube-like formation, but no cytotoxicity against the cells in vitro. The above data suggest that the social isolation stress augmented the tumor-induced angiogenesis probably by up-regulating the angiogenesis-related factors, including TNF-α, VEGF and HGF, and consequently mediating the functions of endothelial cells such as migration, invasion, and tube-like formation.     

Relationship between immediate hypersensitive reactions by buckwheat ingestion and specific IgE for rice in subject with positive IgE-RAST for buckwheat]

IgE-mediated mechanisms are important in immediate hypersensitive reactions (IHR) to buckwheat. However, a part of subjects with high IgE for buckwheat show no IHR to buckwheat ingestion. Inspite of cross-allergenicity between buckwheat and rice, rice ingestion rarely induces IHR even in subjects with high IgE for rice unlike buckwheat-induced IHR. We speculated that there were some relationships between the presence of IHR to buckwheat and recognition of cross-allergenic determinants on buckwheat components with rice components. We examined IgE-RAST for rice in 58 subjects with positive IgE-RAST for buckwheat. IgE-RAST for Dermatophagoides pteronyssinus (Dp), egg white and cow's milk as unrelated antigens with rice were also assessed for a comparison. Subjects (n = 33) without IHR to buckwheat showed higher IgE-RAST values for rice than those (n = 25) with IHR, whereas there were no differences in IgE-RAST values for Dp, egg white and cow's milk between two groups with and without IHR. IgE-RAST values for buckwheat showed significant close correlations to those for rice in subjects without IHR to buckwheat but not in those with IHR. There were no significant correlations between IgE-RAST values for buckwheat and for Dp, egg white or cow's milk in both groups with and without IHR. These results suggested that the IgE from subjects without IHR to buckwheat recognized cross-allergenic determinants with rice on the buckwheat components.