Ovarian cancer presenting as a metastasis to a trocar tract used for a gasless lift-laparoscopy to resect a benign ovarian cyst: an unusual case report.

BACKGROUND: Metastasis to a trocar tract (port-site metastasis, PSM) is an uncommon but serious complication that possibly compromises the prognosis of cancer patients treated laparoscopically. CASE: A 42-year-old Japanese woman had a 20-cm benign right ovarian cyst resected using gasless lift-laparoscopy. Five years and eight months postoperatively, she noticed a three-cm subcutaneous tumor involving the trocar tract. She was also found to have a pelvic mass and an exploratory laparotomy revealed left ovarian cancer. Based on the histopathological findings, the subcutaneous tumor was diagnosed as a metastasis from the ovarian cancer. CONCLUSIONS: This case suggested that PSM could occur without direct or indirect wound contamination during laparoscopic surgery.

Metastatic gastric cancer mimicking an advanced cervical cancer: a case report.

BACKGROUND: Metastasis to the uterine cervix from non-gynecologic neoplasms is rare. However, metastatic tumors sometimes precede the diagnosis of a primary tumor, and may lead to diagnosis of the primary tumor. CASE: A 50-year-old woman was referred to us complaining of increasing right flank pain. Computed tomography scan demonstrated an enlarged uterus with right-sided hydronephrosis and hydroureter. Cervical cytology revealed adenocarcinoma. She was considered to have a Stage IIIB cervical adenocarcinoma. Although no cervical lesion was seen colposcopically, histopathology from biopsies of the uterine cervix revealed poorly differentiated adenocarcinoma infiltrating around the normal endocervical glands. A metastasis from the gastrointestinal tract was suspected. The patient underwent gastroscopy and was found to have Borrmann type IV gastric cancer. Biopsies confirmed a poorly differentiated adenocarcinoma with signet ring cells. CONCLUSION: Physicians should bear in mind that metastatic tumors may precede the diagnosis of a primary tumor and could manifest by mimicking advanced cervical cancer.

Chronic treatment with interleukin-1 beta induces coronary intimal lesions and vasospastic responses in pigs in vivo. The role of platelet-derived growth factor.

Studies in vitro have suggested that inflammatory cytokines may play an important role in the pathogenesis of atherosclerosis. However, little is known about their effects in vivo. Thus, the present study was designed to determine in vivo what histological and functional changes may be induced by chronic treatment with IL-1 beta, one of the major inflammatory cytokines, and also to clarify what mechanisms are involved in those changes. Under aseptic conditions, proximal segments of the left porcine coronary arteries were gently wrapped with cotton mesh absorbing Sepharose beads either with or without recombinant human IL-1 beta. From 1 to 4 wk after the operation, coronary vasospastic responses to intracoronary serotonin or histamine were noted at the IL-1 beta-treated site but not at the control site. Histologically, intimal thickening was greater at the IL-1 beta-treated site than at the control site. Those functional and histological changes induced by the chronic treatment with IL-1 beta were significantly inhibited by the simultaneous treatment with a neutralizing antibody to either IL-1 beta or PDGF. These results indicate that chronic treatment with Il-1 beta induces coronary intimal lesions and vasospastic responses in porcine coronary arteries in vivo and also suggest that these changes are substantially mediated by PDGF.

Tyrosine kinase inhibitor suppresses coronary arteriosclerotic changes and vasospastic responses induced by chronic treatment with interleukin-1 beta in pigs in vivo.

We recently demonstrated that chronic treatment with IL-1 beta induces coronary arteriosclerotic changes and vasospastic responses to autacoids in pigs in vivo and that those responses are importantly mediated by PDGF. The receptors for PDGF and other major growth factors are known to have tyrosine kinase activity. We therefore investigated the effects of a selective tyrosine kinase inhibitor, ST 638, on those responses induced by IL-1 beta in our swine model. Intimal thickening and coronary vasospastic responses to serotonin and histamine were induced at the site of the coronary artery where IL-1 beta was chronically and locally applied. These responses were significantly suppressed in a dose-dependent manner by cotreatment with ST 638. In addition, ST 494, which is an inactive form of ST 638, did not inhibit those responses. The treatment with ST 638 alone did not affect the coronary vasoconstricting responses to the autacoids. Immunoblotting using an antibody to phosphotyrosines confirmed the inhibitory effects of ST 638 on the tyrosine phosphorylations induced by IL-1 beta. These results thus suggest that tyrosine kinase activation may play an important role in mediating the effects of IL-1 beta, while also suggesting that ST 638 has an inhibitory effect on the arteriosclerotic changes and vasospastic responses to autacoids in our swine model in vivo.

[Video-assisted thoracoscopic lobectomy for lung cancer patients with severe incomplete fissures or emphysema; intralobar no-touch access technique (INTACT)].

OBJECTIVES: Severe air leak after lobectomy is one of well-known complications in lung cancer patients with severe emphysema or incomplete fissure. A developing air leak or persistent air leak in those patients may occur postoperative pneumonia or empyema. MATERIALS AND METHODS: For those patients, we have operated with intralobar no-touch access technique (INTACT) assisted with thoracoscopy. INTACT is a surgical procedure of lobectomy without interlobar access, a procedure of prior dissection of lobar bronchus and stapling all the fissure. From January 2004 to December 2005, we performed surgery in 227 patients with primary lung cancer, and 12 patients were performed with lobectmy with INTACT among them. Mean age was 68 +/- 7 years. RESULTS: Distribution of lobectomies was as follows: right upper lobe, 7 patients; right upper lobe and S6 segment, 1; right middle lobe and right lower lobe, 1; left upper lobe, 2; left lower lobe, 1. The length of major skin incision was 5-15 (average 9.2 +/- 3.0) cm. Pathological stage were IA in 3 patients, IB in 7, IIIA in 1 and IIIB in 1. The backgrounds of those patients were severely emphysema in 7 patients, incomplete fissure in 2, both emphysema and incomplete fissure in 2 and emphysema and huge sized tumor in 1. The postoperative air leak stopped 1.8 +/- 0.8 days. CONCLUSIONS: For carefully selected patients video-assisted thoracoscopic lobectomy by INTACT for lung cancer patients with emphysema or incomplete fissure could be a safe and valuable procedure.

Isolation and characterization of thermostable chitinases from Bacillus licheniformis X-7u.

Four kinds of thermostable chitinase were isolated from the cell-free culture broth of Bacillus licheniformis X-7u by successive column chromatographies on Butyl-Toyopearl, Q-Sepharose, and Sephacryl S-200. We named the enzymes chitinases I(89 kDa), II(76 kDa), III(66 kDa) and IV(59 kDa). Chitinases II, III and IV possessed extremely high optimum temperatures (70-80 degrees C), showing remarkable heat stability. Chitinases II, III and IV produced (GlcNAc)2 and GlcNAc from colloidal chitin and chitinase I predominantly produced (GlcNAc)2. The action pattern of chitinase I on PN-(GlcNAc)4 also showed a stronger propensity to cleave off the (GlcNAc)2 unit from the non-reducing end than the other three chitinases. Chitinases II, III and IV catalyzed a transglycosylation reaction that converted (GlcNAc)4 into (GlcNAc)6.

The role of fibroblast growth factor-2 in the vascular effects of interleukin-1 beta in porcine coronary arteries in vivo.

OBJECTIVE: We recently demonstrated that chronic treatment with interleukin-1 beta (IL-1 beta), a major inflammatory cytokine found in atherosclerotic lesions, induces coronary arteriosclerotic changes and vasospastic responses to serotonin and histamine in pigs in vivo and that those responses are partially mediated by platelet-derived growth factor (PDGF). This study was designed to examine, first, whether the effects of IL-1 beta are also partially mediated by fibroblast growth factor-2 (FGF-2), which is another important growth factor in atherosclerotic lesions, and, secondly, whether chronic treatment with FGF-2 per se also induces histological and functional changes in porcine coronary arteries in vivo. METHODS: Porcine coronary arteries were aseptically wrapped with cotton mesh absorbing IL-1 beta with or without neutralizing antibody to FGF-2. In a separate series of experiments porcine coronary arteries were chronically treated with FGF-2 itself in the same manner. Coronary vascular responses in vivo and histological changes were examined 2 weeks after the operation. RESULTS: Coronary vasospastic responses to serotonin and histamine and neointimal formation were induced at the site of the coronary artery where IL-1 beta was chronically and locally applied. These responses were significantly suppressed by co-treatment with a neutralizing antibody to FGF-2 but not by that with non-immune IgG. Immunostaining revealed the presence of FGF-2 in the endothelial cells, the thickened intima and the media at the IL-1 beta-treated site. Furthermore, chronic treatment with FGF-2 also induced coronary vasospastic responses to serotonin and histamine and neointimal formation. CONCLUSIONS: These results suggest that the vascular effects of IL-1 beta may also be mediated by FGF-2 in our swine model and that chronic treatment with FGF-2 also causes coronary arteriosclerotic changes and vasospastic responses in vivo.

Spinocerebellar degenerations in Japan: a nationwide epidemiological and clinical study.

A nationwide survey of patients in Japan with spinocerebellar degenerations (SCD), including SDS and SND, was conducted from 1988 to 1989. The survey consisted of two parts. The first revealed that the estimated total number of patients with SCD in Japan was 5,050 (range: 4,100-6,000) with an estimated prevalence of 4.53 per 100,000 in 1987. The second part investigated the neurological and functional status of patients with SCD. The percentages of those belonging to each subtype of SCD were: OPCA; 34.4%, LCCA; 15.2%, MHCA; 12.6%, HHCA; 7.5%, SDS; 7.0%, HSP; 3.9%, DRPLA; 2.5%, FA; 2.4%, MJD; 2.0% and SND; 1.5%. Compared with European epidemiological studies Japan had a higher proportion of non-hereditary types of SCD. Various clinical features of SCD subtypes were compared grouped by pathological lesion and heredity. HHCA and LCCA: cerebellar ataxia predominated in all stages, and neurological signs other than cerebellar ataxia were rare. MHCA, DRPLA and MJD: in the early phase ataxia was the most common symptom in MHCA, the AC form of DRPLA and MJD, but ataxia was less common and chorea or epilepsy were often observed in ME and PH forms of DRPLA. Other frequently observed clinical features were parkinsonian rigidity in MHCA, abnormal movements and posture in DRPLA and MJD, and disturbances of eye movements in MHCA, the AC form of DRPLA and MJD. OPCA, SDS and SND: dominant clinical features were cerebellar ataxia in OPCA, autonomic disturbance in SDS, and parkinsonian rigidity in SND. FA and HSP: both were rare in Japan. Clinical features related to supra-supinal lesions were frequently observed in FA. Functional status of SCD: the severity of illness was significantly associated with the level of independence in each item of ADL. Activities not requiring dynamic balance were performed independently for a longer period than those requiring dynamic balance. Among SCD subtypes, functional prognosis was poorest in non-hereditary, multi-systemic types (OPCA, SDS and SND) followed by hereditary multi-systemic types (MHCA, DRPLA and MJD), and better in spinal types (FA and HSP) and cerebellar types (HHCA and LCCA).

Genetic linkage study of Duchenne muscular dystrophy and hemophilia A.

We studied a family in which three boys had both Duchenne muscular dystrophy and hemophilia A. Seven other males had only hemophilia A. Genetic linkage study showed a maximum likelihood estimate of recombination frequency of these two X-linked recessive traits of 0.16 with 95% confidence limits of 0.045 and 0.50. The loci of the two diseases may be closer than previous estimates.

Spinocerebellar ataxia type 6. Molecular and clinical features of 35 Japanese patients including one homozygous for the CAG repeat expansion.

Spinocerebellar ataxia type 6 (SCA6) is a newly classified autosomal-dominant cerebellar ataxia (ADCA) associated with CAG repeat expansion. We screened 111 patients with cerebellar ataxia for the SCA6 mutation. Of these, 35 patients were found to have expanded CAG repeats in the SCA6 gene, indicating that second to SCA3, SCA6 is the most common ADCA in Japan. Expanded alleles ranged from 21 to 29 repeats, whereas normal alleles had seven to 17 repeats. There was no change in the CAG repeat length during meiosis. The age at onset was inversely correlated with the repeat length. The main clinical feature of the 35 patients with SCA6 was slowly progressive cerebellar ataxia; multisystem involvement was not common. The 35 patients included nine cases without apparent family history of cerebellar ataxia. The sporadic cases had smaller CAG repeats (21 or 22 repeats) and a later age at onset (64.9 +/- 4.9 years) than the other cases with established family history. We also identified one patient who was homozygous for the SCA6 repeat expansion. The homozygote showed an earlier age of onset and more severe clinical manifestations than her sister, a heterozygote carrying an expanded allele with the same repeat length as the homozygote. This finding suggests that the dosage of the CAG repeat expansion plays an important role in phenotypic expression in SCA6.

Synthesis of N-acetylglucosamine-modified ara-C and its effect on ovarian cancer cells.

1-beta-D-Arabinofuranosylcytosine (ara-C) was modified by reaction of tetra-N-acetylchitotetraose ((GlcNAc)4) using the transglycosylation activity of thermostable chitinase (EC 3.2.1.14) from Bacillus licheniformis X-7u. The structure of the modified ara-C was determined to be either beta 1-3'- or beta 1-5'-linked GlcNAc-ara-C or (GlcNAc)2-ara-C. The total yield of these glycosylated ara-Cs was about 10%. GlcNAc-ara-C and (GlcNAc)2-ara-C depressed the growth of G-401 cancer cells, while 5-O-beta-D-galactopyranosyl-beta-D-arabinofuranosylcytosine (Gal-ara-C) had no effect on G-401 cells.

Antitumor effect of the angiogenesis inhibitor agm-1470 and its combination effect with tamoxifen in dmba induced mammary-tumors in rats.

AGM-1470 is known to inhibit angiogenesis. It inhibited the mitogen induced cell growth in microvascular endothelial cells in vitro. Ten ng/ml AGM-1470 completely inhibited both basic fibroblast growth factor induced cell growth, and interleukin-4 induced cell growth. In an in vivo study, we invesigated the antitumor effect of AGM-1470 in dimethylbenz[a]anthracene (DMBA) induced rat mammary tumors. A dose dependent growth inhibition was seen between administered doses 10 mg/kg and 40 mg/kg. Forty mg/kg 2-week administration of AGM-1470 totally suppressed the tumor growth. In addition, an additive inhibitory effect of AGM-1470 with an antiestrogen tamoxifen was found (p<0.01). It is suggested that the combination of these two drugs might be effective for the treatment of breast cancer.

Production of minor lymphocyte stimulatory-1a antigens from T cell subsets.

T cell subsets that produce minor lymphocyte stimulatory (Mls) antigens were analyzed using mixed lymphocyte reaction (MLR) in vitro or clonal elimination assay in vivo. When lymph node T cells from B10.BR(Mls-1b) mice were stimulated with various T cell subsets from AKR (Mls-1a) mice in the presence of B10.BR antigen presenting cells (APC), proportions of Mls-1a reactive T cell blasts (V beta 6+, V beta 8.1+) increased. The stimulatory potency of CD8+ T cells was higher than that of CD4+ T cells. Furthermore, among either CD8+ or CD4+ T cell subset, CD44+ T cells appeared to produce larger amounts of Mls-1a antigens than CD44- T cells. More marked difference was demonstrated, when stimulator AKR T cells were being activated by immobilized anti-T cell antigen receptor (TCR) antibody during MLR. Thus, AKR T cells appeared to produce large amounts of Mls-1a antigens on appropriate stimulations. These findings were confirmed by the semiquantitative analysis of mRNA levels of MTV-7 in the AKR T cell subsets. When CD8+CD44+ T cells from (AKR x B10.BR)F1 mice were injected intravenously into [B10.BR-->B10.BR] syngeneic bone marrow (BM) chimeras 1 week after BM reconstitution and proportions of V beta 6+ T cells were quantitated 7 weeks later, significant clonal elimination of V beta 6+ T cells was induced among both thymocyte population and lymph node T cell population in a dose-dependent manner of the inoculated F1 T cells. Inoculation of CD8+CD44-F1 T cells eliminated V beta 6+ T cells less efficiently from lymph node T cells and inoculation of CD4+F1 T cells induced no significant clonal elimination of the V beta 6+ T cells. The present findings demonstrate clearly that CD8+CD44+ T cells represent the cells producing large amounts of Mls-1a antigens and inducing clonal elimination of V beta 6+ T cells in vivo.

Delayed clearance of zymosan-induced granuloma and depressed phagocytosis of macrophages with concomitant up-regulated kinase activities of Src-family in a human monocyte chemoattractant protein-1 transgenic mouse.

A human monocyte chemoattractant protein-1 (hMCP-1) transgenic mouse (Tgm) line which constitutively produces a large amount of hMCP-1 (7-13 ng/ml in the serum) was established. Although expression of the transgene was detected in various tissues, an accumulation of macrophages (Mphi) was seen in only lymphoid organs which might be attributed to the high concentration of hMCP-1 in these organs. A reduced phagocytosis by peritoneal Mphi in vivo and a delayed clearance of granulomas in the liver following zymosan administration were observed in these Tgm. However, peritoneal exudate cells (PEC) from Tgm exhibited normal in vitro phagocytic activity and nitric oxide (NO) production upon stimulation with IFN-gamma as compared with those from non-Tgm. In addition, high activities of src-family protein tyrosine kinases (PTK), Fgr and Hck, were also noted in the peritoneal resident cells from Tgm, whereas the level of mitogen-activated protein kinase (MAPK) activity was almost the same as that of non-Tgm. It was suggested that the low functional activities of Tgm Mphi seen in vivo were attributed to down-regulation of the unique transducing system of hMCP-1 signals under the influence of a high concentration of the hMCP-1. It seemed that the depressed functions were recovered when the peritoneal cells were released ex vivo from such a high hMCP-1 environment.

Effects of non-major histocompatibility antigens on acute graft-versus-host reaction after allogeneic bone marrow transplantation.

In the present study using an experimental BMT system we analyzed the effects of disparity at non-MHC Ag including minor lymphocyte stimulatory-1a (Mls-1a) Ag on the acute GVH reaction (GVHR) induced by MHC class I Ag. Mismatch at MHC (class I) Ag alone did not induce clinically detectable acute GVHR in this model. However, BMT mice prepared with a combination of both class I and non-MHC Ag mismatches showed signs of clinical GVHR and various cytokines were produced by the spleen cells at an early stage (4 days) after BMT. Although no clinical GVHR was detected in BMT chimeras prepared with a non-MHC mismatched but MHC matched combination, large amounts of various cytokines were secreted by spleen cells. Cytokine production in the latter two kinds of chimeras paralleled the increase of Mls-1a reactive Vbeta6+ T cells in the host spleen. Marked cytokine production induced by Mls-1a Ag was confirmed by MLR. Thus, these cytokines appeared to be produced by T cells responding to Mls-1a (ie Vbeta6+ T cells) and to augment the T cell responses to MHC class I which resulted in clinically detectable GVHR in chimeras prepared with the combination mismatched at both MHC class I and non-MHC loci.

Complete-type DiGeorge syndrome treated by bone marrow transplantation.

BMT was carried out on a patient with DiGeorge syndrome who suffered recurrent infections after birth. At 13 months of age, 8.0 x 10(8)/kg of bone marrow nuclear cells were infused from an HLA-identical sibling using only anti-thymocyte globulin to prevent rejection. Donor DNA was not detected on microsatellite polymorphism by PCR. At 19 months of age, a second BMT from the same donor was carried out using busulfan and cyclophosphamide as conditioning. DNA examination of bone marrow showed chimerism at day 18 and complete donor origin at day 28. Seven months post-BMT, the numbers of CD3-, CD4- and CD8-positive cells were in the normal range. BMT is thus an effective therapy for DiGeorge syndrome.

Tyrosine hydroxylase and tryptophan hydroxylase activities and its cofactor biopterin level in brain regions of the rolling mouse: The influence of thyrotropin releasing hormone.

Biopterin, the cofactor for tyrosine hydroxylase and tryptophan hydroxylase, was decreased in caudate nucleus, hypothalamus and cerebellum of the rolling mouse. Though there were not significant differences of tyrosine hydroxylase and tryptophan hydroxylase activities between the rolling and normal control mouse in the hypothalamus, the rolling showed significant increase of biopterin concentration and tyrosine hydroxylase activity after administration of thyrotropin releasing hormone (TRH). These results suggest that ataxic gait of the rolling mouse may be partly due to some abnormalities of catecholaminergic neurons, especially noradrenergic neurons, and that TRH may improve the abnormalities of catecholaminergic neurons. The changes of biopterin concentration by TRH administration indicate that biopterin may be a regulatory factor in catecholamine biosynthesis.

Influence of graft versus host reaction on the T cell repertoire differentiating from bone marrow precursors following allogeneic bone marrow transplantation.

When lethally irradiated AKR (Mls-1a) mice were reconstituted with bone marrow (BM) cells plus a small number (0.5%) of mature T cells from allogeneic B10.AQR or B10 (Mls-1b) mice and minor GVHR was induced in the recipients, almost complete donor chimerism was accomplished in the early stages after reconstitution. By contrast, in irradiated AKR mice reconstituted with T cell-depleted BM cells alone from B10 or B10.AQR mice, radio-resistant T cells of recipient origin persisted for a relatively long period in peripheral lymphoid tissues. In this paper the influence of residual T cells in the chimeric mice on generation of the T cell repertoire derived from donor BM is discussed. It will be demonstrated that the recipient (AKR) T cells are capable of producing Mls-1a antigens (Ag) after lethal irradiation in vivo. These recipient T cells eventually induce clonal elimination of Mls-1a reactive V beta 6+, V beta 8.1+ and V beta 9+ T cells derived from developing thymocytes of donor BM origin. The Mls-1a reactive T cells are not eliminated in GVHR chimeras in which recipient T cells are absent. However, V beta 5+ T cells reactive to I-E plus Etc-1 Ag are deleted in the chimeras undergoing GVHR. These results indicate that recipient cells which produce tissue-specific antigens (tolerogens) should be taken into consideration when generation of the T cell repertoire of donor origin following allogeneic BM transplantation is investigated.

Production of interleukin-12 and expression of its receptors by murine microglia.

Production of interleukin-12 (IL-12) by cultured murine microglia and astrocytes was examined, by means of ELISA to detect heterodimeric p70 and RT-PCR to analyze the expression of mRNA encoding p35 and p40. Microglia, but not astrocytes, produced IL-12 p70 in response to lipopolysaccharide and interferon-gamma. The microglial cell line, Ra2, produced only p40, but not p35, upon above stimulation. Thus, it is possible that some population of microglia induce helper 1 type T cell response via producing IL-12 in the CNS. Microglia were induced to express mRNA encoding IL-12 receptors which were exclusively expressed in activated T and NK cells.

Multinucleated giant cell formation by microglia: induction by interleukin (IL)-4 and IL-13.

To investigate the cellular origin and the mechanisms of multinucleated giant cell (MGC) formation in the CNS, various cytokines were applied to isolated murine microglia. All the single cytokines failed to induce MGC. However, interleukin (IL)-13 and IL-4 induced microglial MGC formation in the presence of colony stimulating factors, which was inhibited by either anti-IL-13 or anti-fms antibody. T helper 2 (Th2)-derived cytokines can induce MGC formation even in the absence of infectious agents.