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OBJECTIVE We examined the hypothesis that antibiotic treatment with or without gonadotropin releasing hormone agonist (GnRHa) may decrease intrauterine infection with consequent decrease in tissue inflammation, cell proliferation and angiogenesis in human endometriosis. STUDY DESIGN This is a prospective non-randomized observational study. Endometrial/endometriotic samples were collected during surgery from 53 women with endometriosis and 47 control women who were treated with levofloxacin (LVFX, 500 mg, once per os) or GnRHa (1.88 mg/IM for 3 months) before surgery. Endometrial samples were analyzed by broad-range polymerase-chain reaction (PCR) amplification of bacteria targeting V5-V6 region of 16S rRNA gene. Immunohistochemical analysis was performed using antibodies against CD138 (Syndecan-1, a marker of plasma cells), CD68 (marker of macrophages), Ki-67 (cell proliferation marker), and CD31 (vascular cells marker). RESULTS 16S rDNA metagenome assay indicated that treatment with either of LVFX or GnRHa + LVFX significantly decreased some components of major bacterial genera comparing to untreated group. In women with endometriosis, treatment with either of LVFX or GnRHa + LVFX significantly decreased Gardnerella, Prevotella, Acidibactor, Atopobium, Megasphaera, and Bradyrhizobium (p

Nonalcoholic fatty liver disease is a major liver disease that leads to cirrhosis and/or hepatocellular carcinoma in a subset of patients. The mechanism underlying disease progression is largely unknown. p53-binding protein 1 (53BP1) is a DNA damage response protein that rapidly localizes at the site of DNA double-strand breaks. In this study, we investigated nuclear 53BP1-positive foci formation as an indicator of DNA double-strand breaks in human nonalcoholic fatty liver disease liver tissues by immunofluorescence microscopy. A total of 52 liver tissue samples, including 43 nonalcoholic fatty liver disease samples and 9 controls, were studied. Our results show that the number of abnormal 53BP1-positive foci in hepatocytes (defined as three or more discrete nuclear foci and/or large foci greater than 1 μM) was significantly increased in nonalcoholic fatty liver disease patients compared to that in controls, both in nonalcoholic fatty liver (p

Research question Is there any relationship between numbers of FOXP3+ regulatory T-cells (Treg) and occurrence of peritoneal lesions in women with ovarian endometrioma and dermoid cysts? Design Retrospective and prospective case-controlled cohort study. Peritoneal lesions were collected from 27 women with ovarian endometrioma and 25 women with dermoid cysts. Peritoneal fluid was collected from 36 women with ovarian endometrioma and 42 women with dermoid cysts. Tissue expression of Forkhead box P3 (FOXP3), one of the transcription factors of Treg cells, and transforming growth factor-beta (TGF-β) were examined by immunohistochemistry. Interleukin-6 (IL-6) and TGF-β levels in the peritoneal fluid were measured by enzyme-linked immunosorbent assay. Results Ovarian endometrioma cases with coexisting peritoneal lesions were significantly more frequent than dermoid cyst cases with coexistent peritoneal lesions (269/350 [76.9%] versus 74/414 [17.9%]; P Conclusions This study confirms current speculation that endometriosis is related to alteration in Treg cells, causing survival and implantation of ectopic endometrial lesions in women with endometrioma and dermoid cysts. The findings may clarify why only 10% of women in the general population develop endometriosis despite cyclic menstruation with retrograde flow occurring in >90% of women.

Purpose/Objective(s) The cyclooxygenase-2 product prostaglandin E2 (PGE2) promotes tumor growth and metastasis by acting on a family of four G protein-coupled receptors (EP1–4). Recently, an antagonist of PGE2-EP4 (AAT-008) has been newly developed for clinical use. The purpose of this study is to investigate the biological effects of AAT-008 as a radiosensitizer for the treatment of murine colon cancer (CT26WT) in vivo and explore the mechanism using flow cytometry. Materials/Methods CT26WT tumor cells transplanted into the hind legs of BALB/c mice were used. Treatments were carried out when the mean diameter of the tumors reached approximately 10 mm for tumor growth delay assay and 5 mm for flow cytometry (day 0). AAT-008 (0, 3, 10, and 30 mg/kg/day) was administered orally twice a day from day 0 to day 18. The tumor was irradiated at 9 Gy on day 3 for the radiation treatment (RT) group. In the tumor growth assay, twelve mice were used for each group. The three dimensions of each tumor were measured every other day using a caliper and the tumor doubling time was calculated. For the first flow cytometry, the dose of AAT-008 was fixed to 10 mg/kg/day. Administration of AAT-008 and RT dose were the same as in the tumor growth assay. The population of cytotoxic T-cells (CD45+CD8+CD69+: CTL) was investigated using the tumor cells on day 19. For the second flow cytometry, the dose of AAT was fixed to 30 mg/kg/day. AAT-008 was administered from day 0 to day 12. The population of CTL and the ratio of CTL / regulatory T-cells (Foxp3+: Treg) were investigated using the tumor cells on day 13. Results In the tumor growth delay assay, the median tumor doubling times in the 0-, 3-, 10-, and 30-mg group without RT were 3.2, 4.4, 4.4, and 5.7 days, respectively. The times in those dose groups with RT were 6.1, 6.6, 17.3, and 19.8 days, respectively (P Conclusion AAT-008 had a potential of enhancing radiosensitivity in the treatment of colon cancer cells. It was assumed that the optimal dose of AAT-008 with RT was 10 mg/kg/day. It was suggested that AAT-008 stimulated the immune system against the cancer cells.

Background The semidry dot-blot method is a diagnostic procedure for detecting lymph node (LN) metastases using the presence of cytokeratin (CK) in lavage fluid from sectioned LNs. We evaluated 2 novel kits that use newly developed anti-CK-19 antibodies to diagnose LN metastases in breast cancer. Patients and Methods We examined 159 LNs dissected that we sliced at 2-mm intervals and washed with phosphate-buffered saline. The suspended cells in the lavage were centrifuged and lysed to extract protein. This extracted protein was used with a low-power and a high-power kit to diagnose LN metastasis. Diagnoses on the basis of the kits were compared with pathological diagnoses. Results Of the 159 LNs, 68 were assessed as positive and 91 as negative in permanent section examination. Sensitivity, specificity, and accuracy of the low-power kit for detecting LN metastases was 83.8%, 100%, and 93.1%, respectively. Those of the high-power kit were 92.6%, 92.3%, and 92.5%, respectively. Combining the low- and high-power kit results, those for distinguishing macrometastases were 94.5%, 95.2%, and 95.0%, respectively. Diagnosis was achieved in approximately 20 minutes, at a cost of less than $30 USD. Conclusion The kits were accurate, fast, and cost-effective in diagnosing LN metastases without the loss of LN tissue.

BH3-only protein, Bim, is a pro-apoptotic protein that mediates mitochondria-dependent cell death. However, the role of Bim in Helicobacter pylori-associated gastritis remains unclear. This study aimed to assess the cellular localization of Bim and its possible role in H. pylori-induced gastritis. The study was conducted on biopsy specimens obtained from 80 patients who underwent upper gastrointestinal endoscopy (H. pylori-negative: n = 30, positive: n = 50). Association between Bim mRNA expression and severity of gastritis was evaluated and the localization of Bim was examined by immunofluorescence. Bim mRNA expression was positively correlated with the degree of gastritis, as defined by the Sydney system. Immunohistochemical analysis confirmed increased Bim expression in H. pylori-infected gastric mucosa compared with uninfected mucosa in both humans and mice. Bim localized in myeloperoxidase- and CD138-positive cells of H. pylori-infected lamina propria and submucosa of the gastric tract, indicating that this protein is predominantly expressed in neutrophils and plasma cells. In contrast, Bim did not localize in CD20-, CD3-, or CD68-positive cells. Bim was expressed in the mitochondria, where it partially co-localized with activated Bax and cleaved-PARP. In conclusion, Bim is expressed in neutrophils and plasma cells in H. pylori-associated gastritis, where it may participate in the termination of inflammatory response by causing mitochondria-mediated apoptosis in specific leucocytes.

We treated a 45-year-old Japanese woman with primary Sjogren's syndrome (SS) complicated with dermatomyositis (DM) followed by diffuse large B-cell lymphoma. She was admitted to our hospital for further evaluation of fever, weight loss and peritoneal lymphadenopathy. The histological examination of her lymph node revealed diffuse large B-cell lymphoma. The patient was then treated with 8 cycles of R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone) therapy, which resulted in histological and functional improvement of her salivary glands. Of note, the percentage of effector memory Th17-1 (CD3+CD4+CD45RA-CCR7-CXCR3+CCR6+) cells in the peripheral blood was decreased after the R-CHOP treatment. This case suggests that an altered Th17-1 cell subset by B-cell depletion therapy is critical for the improvement of tissue damage in patients with SS, and the case suggests that clinicians should consider measuring the effector memory Th-subsets to predict the disease activity in SS patients.

Summary Studies have typically emphasized one of three major policy alternatives—government (state) ownership, privatization, or user self-governance—to address overuse of “the commons” as a natural resource shared by many competing users. Studies tend to focus on each alternative separately. Government ownership or privatization is usually understood to undermine user self-governing institutional arrangements, while user self-governance has proved to be a very powerful policy alternative in managing the commons in many cases. An important research question arises as to whether a complex policy design can strengthen the competence of user self-governing institutional arrangements. This article defines a complex policy design as one that involves a mix of flexible policy alternatives rather than a rigid alternative to address overuse issues. Drawing on Japan’s irrigation water management experience, this study demonstrates that when a complex policy design is tailored to facilitate user autonomy, it further strengthens user self-governance. The study provides scholars with insight into how self-governing institutional arrangements—which were primarily developed in the existing literature with the government’s role assumed as absent or implicit—could be enhanced when the role is strategically explicit.

Aims Genomic instability has been indicated during the dedifferentiation process from leiomyoma (LM) to leiomyosarcoma (LMS). Previously, we have described that nuclear expression pattern of DNA damage response protein p53-binding protein 1 (53BP1), detected by immunofluorescence, reflects the magnitude of genomic instability during malignancy. Here, we present a case of LMS arising from LM with molecular analysis of 53BP1, which showed transitional magnitude of DNA damage response within a tumor. Methods and results A fifty-year-old female with abdominal mass underwent hysterectomy. Histologically, the tumor consisted of LMS with highly atypical multinucleated giant cells as well as an LM component with transitional atypical spindle cells in the border area. LMS showed diffuse nuclear staining of 53BP1 expression, which has been previously described as high DNA damage response pattern. In contrast, the LM component lacked 53BP1 immunoreactivity and focal expression was observed in transitional lesion. Furthermore, double-labelled immunofluorescence revealed co-localization of 53BP1 with p53 and Ki-67 in the LMS component, which indicated abnormal DNA damage response in proliferative state. Conclusions This study revealed that diffuse-type 53BP1 expression may be beneficial to estimate genomic instability during dedifferentiation from LM to DLMS.

Two methods of substrate scratching pretreatment using diamond powder are employed to control the wall spacing in carbon nanowalls (CNWs). The surface after scraping for undulation has continuous undulant scratches with a number of residual diamond grains exclusively along the scratches, while that after scratching with ultrasonic vibration it shows irregular distributions of residual diamond grains and scratches, depending upon the size of the diamond powder. Nanocrystalline diamond film/CNW composites are obtained with either pretreatment method by microwave plasma-enhanced chemical vapor deposition. With increase of the duration of scratching, the morphology of the deposits changes from CNWs to a film/CNW composite and lastly to CNWs on a film, accompanied by an overall increase in wall spacing. The turn-on field for field emission decreases from 2.1 V/μm without scratching down to 1.2 V/μm with scratching due to suppression of electric field screening between the walls as evidenced by the larger field enhancement factor up to ~ 2700.

We recently demonstrated that IL-18 injections following burn restored IFN-γ production and increased mouse survival after bacterial infection. However, it has yet to be fully elucidated how the IL-18 therapy affects the function of phagocytic cells. We investigated the effect of IL-18 therapy on function and interaction of Kupffer cells and NK cells in burned mice. C57BL/6 mice received a 20% full-thickness burn, followed by multiple injections with IL-18. Although burn-injured mice had decreased expression of IL-18 receptors on the NK/NKT cells 5 days after injury, multiple IL-18 injections restored this expression. IL-18 treatment also augmented Kupffer cell phagocytosis. Although burn decreased the number of CD68(+) Kupffer cells with phagocytic activity, IL-18 treatment partially restored their proportion, and augmented phagocytosis-induced ROS production in CD68(+) Kupffer cells after the injection of heat-killed Escherichia coli. Consistently, IL-18 restored the impaired E. coli killing activity of Kupffer cells of burn-injured mice. Such Kupffer cell activation by IL-18 was abrogated by the deletion of NK cells or IFN-γ. In conclusion, IL-18 therapy in burn-injured mice enhanced function of CD68(+) Kupffer cells via the activation of liver NK cells and augmentation of their IFN-γ production, thereby improving survival after E. coli infection.

Liver Kupffer cells have been suggested to be heterogeneous macrophage lineage cells. We explored this possibility by classifying the mouse Kupffer cells into subpopulations and characterizing them by their phenotype and function.Liver mononuclear cells (MNCs) from C57BL/6 mice were isolated and their phenotypes and functions were analyzed. The effects of clodronate liposomes and gadolinium chloride (GdCl(3)) on Kupffer cells were also investigated.Approximately 25% of liver MNCs were F4/80(+) Kupffer cells. Of these, 46% were CD11b(-)CD68(+), 22% were CD11b(+)CD68(-), and 6% were CD11b(+)CD68(+). CD68(+) cells showed potent phagocytic activity and reactive oxygen species (ROS) production capacity after lipopolysaccharide (LPS) stimulation, whereas CD11b(+) cells did not. CD11b(+) cells showed a strong capacity for the production of cytokines (TNF and IL-12), which was much less prominent in CD68(+) cells. At 24h after LPS or Escherichia coli injection into mice, the proportions of CD11b(+)CD68(-) and CD11b(+)CD68(+) cells increased but that of CD11b(-)CD68(+) cells decreased. The increase in CD11b(+)CD68(+) cells appeared to be derived from the CD11b(+)CD68(-) subset. Although the CD11b(+) cells augmented phagocytic activity after LPS injection, they did not increase ROS production, suggesting their weak lytic activity. Injection of clodronate or GdCl(3) into mice depleted the CD68(+) cells but increased CD11b(+) cells proportionally because CD68(+) cells may phagocytose these toxic reagents and undergo apoptosis. GdCl(3)-treated mice also consistently increased serum TNF after LPS challenge.Two F4/80(+) Kupffer cell subsets may exist, a CD68(+) subset with phagocytic activity and a CD11b(+) subset with cytokine-producing capacity.

Field emission performance of nitrogen-incorporated vertically-aligned graphene layers, so called “carbon nanowalls (CNWs)”, is found to depend upon their electrical conduction properties. The CNW samples with n-type conduction exhibit near-ideal Ohmic contacts with various metals such as Cu, Ti, and Au, regardless of the work function of the metals. The high resistivity CNWs for lower deposition temperatures (TD) show semiconducting behavior in the Arrhenius plots, while the low resistivity CNWs for higher TD show semi-metallic behavior. The emission turn-on field versus TD has a very similar trend to the bulk resistivity versus TD, and is reduced down to about 3 V/μm when the resistivity reaches a minimum (3.0 × 10− 3 Ω cm). The lower turn-on field for semi-metallic CNWs is attributed to an upward shift of the emission level, which is responsible for a decrease in the surface potential barrier height for emission.

RET oncogene rearrangement is a well-known molecular alteration observed in papillary thyroid cancer (PTC). RET rearrangement is the commonest oncogenic alterations in Chernobyl-related PTC; nearly all to RET/PTC1 or RET/PTC3, resulting from paracentric inversion of chromosome 10. Other less common variants, usually forming as a result of interchromosomal translocation, occur in an extremely limited number of cases. Most often such translocations have been found in radiation-induced PTC. Here, we report the presence of RET oncogene amplification, a new type of RET cytogenetic alteration, in human thyroid cancers. RET amplification was correlated with radiation-associated, high-grade malignant potency, and p53 overexpression, suggesting genomic instability. We also examined p53-binding protein 1 (53BP1) expressions with immunostaining to clarify the presence of genomic instability in thyroid cancers. In thyroid tumors, the level of 53BP1-focus formation was associated with malignant transformation and high-grade malignant potency as well as RET amplification. RET amplification might be induced by a high level of genomic instability with progression of thyroid carcinogenesis and, subsequently, be associated with radiation-induced and/or high-grade malignant cases.

Primary pulmonary artery sarcomas (PASs) are rare and lethal tumors. They are easily misdiagnosed as chronic pulmonary embolism, mediastinal mass or tumor emboli, which delay a proper treatment. Although the advanced technologies are now increasingly being used, their diagnosis is usually hard to establish preoperatively at the present time. We report here a case of a 68-year-old female with PAS with lung metastases, who firstly presented with symptoms of common cold and anemia. Although a PAS had been suspected, the final diagnosis of pulmonary intimal sarcoma was made only postoperatively by histological and immunohistochemical examination. The patient died 8 months after the operation because of tumor growth progression, despite adjuvant chemotherapy and radiation therapy. Although pulmonary intimal sarcomas are usually of poorly differentiated mesenchymal malignancy, most reported cases are immunohistochemically positive for vimentin, alpha-smooth muscle actin (SMA), and/or desmin, therefore resembling leiomyosarcomas. However, the diagnosis of leiomyosarcoma should not be made on the basis of immunostains in the absence of typical morphologic features, and PAS, like the present case, should be more appropriately classified as intimal sarcoma according to the new WHO Classification of Tumours of Soft Tissue and Bone published in 2002.

Recent reports suggest that bone marrow might serve as a source of skin progenitor cells. Graft-versus-host disease (GVHD) is a systemic disease that involves the skin, the liver, and the gastrointestinal tract, and contributes to transplant-related morbidity and mortality. To evaluate whether donor-derived bone marrow cells participated in its pathophysiology, we correlated the severity of GVHD in skin from sex-mismatched recipients with the percentage of donor-derived keratinocytes using fluorescence in situ hybridization (FISH) for detecting the Y chromosome. Y-positive signals were observed in female epidermis in both keratinocytes and lymphocytes. These results support the notion that donor-derived hematopoietic stem cells may contain pluripotent stem cells. Furthermore, there was a strong correlation between the frequency of Y-positive keratinocytes and the histopathologic grade of GVHD (p = 0.004), as well as with the number of Y-positive lymphocytes infiltrating the epidermis (p = 0.005), suggesting a role for donor-derived keratinocytes in the pathophysiology of GVHD. However, no clusters of Y-positive keratinocytes were found in the epidermis, and no correlation was observed between the number of Y-positive keratinocytes and time course of GVHD after transplantation. We hypothesize that donor-derived stem cells may play a role in the regeneration of damaged keratinocytes in GVHD.

The aims of this study were to elucidate the molecular mechanism by which growth hormone (GH) excess is anti-fibrotic in vitro and in vivo model. The in vivo model GH excess showed a significant increase of relative wall thickness with no concomitant disturbance of cardiac diastolic function. Western blot for extracellular matrix (ECM) structural proteins showed minimal change in the GH treatment group, compared to an Angiotensin II (Ang II) subpressor dose group. In cultured cardiac fibroblasts, we investigated the abundance of ECM proteins, phosphorylation of p38 mitogen-activated protein kinase (MAPK), and transforming growth factor-beta (TGF-beta)-specific transcriptional activity. GH down-regulated the expression of PAI-1 and fibronectin proteins activated by TGF-beta. In reporter assays, GH, but not insulin-like growth factor-1 (IGF-1), reduced TGF-beta-specific transcriptional activity. Moreover, GH markedly down-regulated TGF-beta-induced phosphorylation of p38 MAPK. These results demonstrated that a chronic excess of GH have an anti-fibrotic effect on cardiac remodeling, probably through a down-regulation of TGF-beta signaling via de-phosphorylation of p38 MAPK.

A high incidence of skin cancers has been noted around the Semipalatinsk Nuclear Testing Site (SNTS) in Kazakhstan. Recently, basal cell carcinoma (BCC) susceptibility genes, human homolog of the Drosophila pathed gene (PTCH), and the xeroderma pigmentosa group A-complementing gene (XPA), have been cloned and localized on chromosome 9q22.3. To clarify the effect of low-dose irradiation on the occurrence of BCC, we used microdissection and polymerase chain reaction to identify loss of heterozygosity (LOH) at 9q22.3 using BCC samples obtained from this region. Ten Japanese samples were analyzed as controls. LOH with at least 1 marker was identified in 5 of 14 cases from around SNTS, whereas only 1 case with 1 marker was identified among the 10 Nagasaki cases. The total number of LOH alleles from SNTS (8 of 45) was significantly higher than the number from Nagasaki (1 of 26) (P = 0.03). The higher incidence of LOH on 9q22.3 in BCC from around SNTS suggests involvement of chronic low-dose irradiation by fallout from the test site as a factor in the cancers.

In the tumor tissue of papillary thyroid carcinoma (PTC) developed in an externally irradiated patient, we identified an undescribed yet structural mutation which comprised the juxtapositioning of the 5′ portion of an rfp gene upstream of the fragment encoding the tyrosine kinase domain of RET. At the genomic level, the formation of a chimeric gene occurred as a result of balanced chromosomal translocation of the short arm of chromosome 6 and long arm of chromosome 10, t(6;10)(p21.3;q11.2). The presence of the mutation was confirmed by extensive molecular, cytogenetic, and functional analysis. The product of the fusion gene, termed Δrfp/RET , was able to form homodimers as demonstrated by a yeast two-hybrid system, thus being in compliance with an essential condition of the constitutive activation of ret/PTC tyrosine kinases. In addition, the newly revealed rearrangement was shown to be tumorigenic, as NIH 3T3 cells stably transfected with a mammalian expression vector encoding full-length cDNA of the Δrfp/RET yielded rapidly growing tumors in immunocompromised mice. The finding provides an additional line of evidence of facilitated susceptibility of the ret protooncogene to structural mutations in irradiated human thyroid cells and pathogenic role of the ret rearrangements in human papillary thyroid cancer.

A total of 459 nuclear tests were officially conducted at the Semipalatinsk Nuclear Test Site of Kazakhstan during the period from 1949 to 1989. The exposure to radioactive fallout could affect the population health. The aberrant activation of Wnt pathway elements is linked to thyroid tumorigenesis. The activated Wnt signaling stabilizes β-catenin and results in transactivation of target genes including cyclin D1. To investigate the involvement of the Wnt/β-catenin pathway in thyroid carcinomas around the Semipalatinsk Nuclear Test Site, we compared immunohistochemical results for β-catenin and cyclin D1 expression in chronic thyroiditis, follicular adenoma and papillary carcinoma. This study revealed a higher prevalence of both aberrant β-catenin expression and cyclin D1 overexpression in papillary thyroid cancers around the Semipalatinsk Nuclear Test Site than sporadic cases. The analysis of the alteration of the Wnt signaling-related molecules in thyroid cancer around the Semipalatinsk Nuclear Test Site may be important to gain an insight into radiation-induced thyroid tumorigenesis.

Small cell carcinoma of the endometrium (SCCE) is extremely rare. Previous reports indicate that SCCE frequently shows systemic spread and has a poor prognosis. Beta-catenin has been shown to be a key downstream effector of the Wnt signaling pathway, which regulates cell growth and survival. Decreased membranous expression of beta-catenin in cancers correlates with poor prognosis and is associated with dissemination of tumor cells and the formation of metastases. Recently, some different investigators demonstrated aberrant beta-catenin accumulation in neuroendocrine tumors arising in different organs, suggesting a role for the Wnt/beta-catenin signaling pathway during neuroendocrine tumorigenesis. Here, we report a new case of SCCE associated with peritoneal spreading and aggressive course; the patient died one month after surgery. This study also aimed at assessing the involvement of the Wnt signaling pathway in this rare neuroendocrine tumor. Interestingly, both intense nuclear beta-catenin accumulation and cyclin D1 immunoreactivity were restricted to carcinoma cells invading lymphatic vessels. However, mutation analysis failed to demonstrate any mutation in exon 3 of the beta-catenin gene or exon 15 of the APC gene in the present case. Although the mechanism of nuclear accumulation of beta-catenin is still unknown, the heterotopic nuclear localization of beta-catenin may play a role in the tumor invasion process and, subsequently, may be associated with the aggressive behavior of SCCE.

The purpose of this study was to evaluate the healing effect of interleukin-11 (IL-11) on acetic acid-induced gastric ulcer in rats. Gastric ulcers were induced in male Wistar rats by applying acetic acid to the fundus of the stomach. Recombinant human interleukin-11 (rhIL-11 100 microg/kg/twice daily, subcutaneously) was administered starting on the 2nd day before ulcer induction up through the 7th day after ulcer induction. Control rats were injected with bovine serum albumin. At 12 hours and 7 days after ulcer induction, the animals were sacrificed, and the ulcer index, proliferating cell nuclear antigen (PCNA) expression, and IL-11alpha receptor expression in the gastric tissues were studied. The ulcer index of the rhIL-11-treated rats was significantly lower than that of the control rats at the 7th day. The expression of PCNA as evaluated by Western blotting and immunohistochemistry, was enhanced in both the mucosal proliferative zone and proper muscle layer of the rhIL-11-treated rats in comparison with that in the control rats. IL-11alpha receptor expression was observed in the mucosal neck cells of the rhIL-11-treated rats and control rats. These findings suggest that IL-11 accelerates ulcer healing by inducing the proliferation of mucosal and muscular cells.

Organisms respond to infection in a complex manner involving bidirectional interactions between the neuroendocrine and immune systems. Many of the bioactive endocrine/immune factors are synthesized in a precursor form and are expected to be activated by prohormone convertases (PCs). Since patients with both type 1 and type 2 diabetes have an increased incidence and severity of infections, we hypothesized that in a condition of hyperglycemia, these processing enzymes would be activated in an immune tissue, the spleen. To test this hypothesis, we treated rats with intraperitoneal streptozotocin (STZ) (50 mg/kg/day) daily for 5 days and measured splenic PC1 and PC2 mRNA by ribonuclease protection assay. We found that PC1 mRNA was increased 6.0+/-0.02-fold (P0.05) and PC2 mRNA was increased 1.80+/-0.01-fold (P0.005) in the spleen of rats that received STZ compared to rats that received vehicle. Western blot indicated that the 75-kDa form of PC1 was the only form of PC1 present in the spleen and that this form increased with STZ treatment. Immunohistochemistry revealed that PC1 was found in both the white pulp (T-lymphocytes) and red pulp (monocytes and macrophages) and that its increase in immunoreactivity occurred primarily in the white pulp. PC2 and pro-opiomelanocortin (POMC, a possible splenic substrate for PC1/PC2) immunoreactivity was found predominantly in the red pulp. STZ induced an increase in splenic PC1 and POMC, but not PC2 protein levels. We conclude that in the STZ model of diabetes, splenic PCs are induced, which could lead to an increased activation of many immune-derived hormones. We speculate that this up-regulation of prohormone converting enzymes may be related to the increased infections seen in patients with both type 1 and type 2 diabetes.

Summary We have recently demonstrated that Tie-1 and Tie-2 are expressed in synovial cells from rheumatoid arthritis (RA). To elucidate the possible involvement of Tie receptors in synovial proliferation, we analyzed their expression by immunostaining in five cases of giant cell tumor of tendon sheath (GCTTS), which represents a proliferating lesion of synovial cells. Strong immunoreactivity for both Tie-1 and Tie-2, regardless of the individual patient's profile, was observed in all cases of GCTTS. Six sets of double immunohistochemical stainings for Tie-1/Tie-2 and fibronectin, CD68, or CD34 were carried out to determine the phenotype of Tie-1 and Tie-2-positive tumor components. In these studies, both Tie-1 and Tie-2 immunoreactivity were widely observed in the fibronectin-positive fibroblastic and the CD68-positive histiocytic mononuclear cells, as well as in the osteoclast-like giant cells. In tumor vasculature, Tie receptors were expressed in the CD34-positive endothelial cells possessing proliferating cell nuclear antigen (PCNA) immunoreactivity. We also evaluated the correlation of Tie-1/Tie-2 expression and proliferating cells in GCTTS by using double staining of Tie-1/Tie-2 together with PCNA. Overexpression of PCNA immunoreactivity was frequently found in Tie receptors-positive cells with no obvious differences in the expression pattern of Tie-1 and Tie-2. These findings suggest the possible involvement of Tie receptors in the pathogenesis of GCTTS other than solely via their involvement in angiogenesis and subsequent vascularization. It was demonstrated that Tie-2 immunoreactivity was restricted to the fibroblastic, but not histiocytic, phenotype in RA synovium, suggesting different regulatory control of Tie-2 expression in GCTTS and RA synovium. Overexpression of Tie receptors in GCTTS may imply a biological role for these receptors in synovial proliferation.

Matrix metalloproteinases are considered to play an important role in tumor invasion and metastasis. To elucidate the involvement of MMP-1 in human colorectal carcinoma, we performed immunohistochemical analysis on tissues from 20 colorectal adenomas and 142 colorectal adenocarcinomas, including 27 intramucosal carcinomas and 115 invasive carcinomas. MMP-1 was not expressed in any of the 20 cases of colorectal adenoma examined. In contrast, 108 of 142 cases (76.1%) with colorectal adenocarcinoma showed immunoreactivity for MMP-1 in the carcinoma cells themselves. Expression of MMP-1 was also identified in stromal cells around the carcinoma. We investigated the relationship between pathological features in colorectal carcinoma and MMP-1 immunoreactivity of the tumor cells. MMP-1 expression was less frequent in intramucosal carcinomas and weaker than that in invasive carcinomas (P < .0001). Among the 115 cases of invasive carcinomas, MMP-1 immunoreactivity was significantly correlated with the depth grading of tumor invasion (P < .05), tumor growth pattern (P < .05), the presence of lymphatic invasion (P < .05), venous invasion (P < .05), neural invasion (P < .05), lymph node metastasis (P < .005), hepatic metastasis (P < .05), and increasing stages of Dukes' classification (P < .05). In situ hybridization, using an MMP-1 oligonucleotide probe, confirmed the presence of MMP-1 mRNA in colorectal carcinoma cells themselves. Expression of MMP-1 mRNA was detected by the reverse transcription polymerase chain reaction method in cultured human colorectal carcinoma cell lines and colon carcinoma tissue obtained at surgery. These findings suggest that the expression of MMP-1 is one of the important factors related to tumor invasion and metastasis in colorectal carcinoma.

The effect of dexamethasone (Dex) on the ability to invade type I collagen gel was investigated in two cell lines of oral squamous cell carcinoma (SCC). At concentrations higher than 10(-8) M, Dex significantly suppressed the invasive growth of SCC cells into the gel. The same concentrations of Dex led to a decrease in urokinase type plasminogen activator (u-PA) synthesis and an increase in plasminogen activator inhibitor type 1 (PAI-1) synthesis by SCC cells. These findings suggest that Dex inhibits the invasiveness of SCC cells by decreasing their proteolytic activity.

1738 www.thelancet.com Vol 386 October 31, 2015 3 Moulder JE, Cohen EP. Renal dysfunction after total body irradiation: dose-eff ect relationship: in regard to Kal and van Kempen-Harteveld. Int J Radiat Oncol Biol Phys 2007; 67: 319. 4 Sera N, Hida A, Imaizumi M, Nakashima E, Akahoshi M. The association between chronic kidney disease and cardiovascular disease risk factors in atomic bomb survivors. Radiat Res 2013; 179: 46–52. 5 Go AS, Chertow GM, Fan D, McCulloch CE, Hsu CY. Chronic kidney disease and the risks of death, cardiovascular events, and hospitalization. N Engl J Med 2004; 351: 1296–305. Tumor Tissue Bank for future analysis. Samples from 485 atomic bomb survivors have been collected so far, and the high quality and quantity of DNA and RNA extracted from the samples enable genomic analysis. We have previously reported a higher incidence of multiple primary cancers in atomic bomb survivors, particularly in those who were youngest at the time of and closest to the explosions, and others have reported a higher risk of myelodysplastic syndrome and multiple cancers in atomic bomb survivors. The Nagasaki Atomic Bomb Survivors’ Tumor Tissue Bank will help radiation research, promote worldwide collaboration, and enhance our understanding of the late-onset eff ects of radiation on carcinogenesis at the molecular level. Use of the tissue bank could provide information for the prediction, prevention, and treatment of cancer in both atomic bomb survivors and the individuals affected by the Fukushima nuclear power plant incident.

s / Journal of Reproductive Immunology 101–102 (2014) 40–60 45 and pNK. No correlation was identified between uNK and pNK. Conclusion: Peripheral and uterine NK did not correlate within our study population. Although RM patients showed higher levels of pNK and uNK significance were not achieved. However, the study subgroups were small and different methods were applied to identify pNK and uNK. Therefore, further data are needed to analyze the role of NKs in the pathophysiology of RM and RIF. http://dx.doi.org/10.1016/j.jri.2013.12.041

Background Utilization of formalin-fixed, paraffin-embedded (FFPE) archival tissue, which is the most common form of tissue preservation in routine practice, for cytogenetic analysis of microarray comparative genomic hybridization (aCGH) remains challenging. We searched for predictive factors of FFPE DNA performance on aCGH analysis. Materials and methods Tumor DNA was extracted from 63 various types of FFPE archival tissues (31 of breast cancer, 24 of lung cancer, and 8 of thyroid tumor) followed by aCGH analysis using high-density oligonucleotide microarray. In two cases, tumor DNA from a matched frozen sample and tumor DNA after whole-genome amplification (WGA) from an FFPE sample were also analyzed. The derivative log ratio spread (DLRSpread) was used as a metric to assess the overall quality of each aCGH result. Results The DLRSpread was significantly correlated with the double-stranded DNA ratio of tumor DNA, storage time, and degree of Cy5 labeling (P Conclusions The double-stranded DNA ratio of tumor DNA predicted the performance of DNA from FFPE samples on aCGH analysis. Quality control by these metrics can utilize valuable FFPE archival tissue on aCGH analysis. Disclosure All authors have declared no conflicts of interest.

BackgroundControlledproteolysisis needed for cell migration,angiogenesis, and matrix remodeling during normal wound repair. Coordination among these processes has not been well studied. Ets-l is a transcription factor known to control gene expression of matrix metalloproteases (MMPs) such as MMP-l, MMP-3, MMP-9, and MMP-lO. MMP-I and MMP-3 are deeply involved in angiogenesis and matrix remodeling. MMP-9 and MMP-1O playa pivotal role of epithelial cell migration. The purposeof this study is to evaluateexpressionof Ets-I and its target genes of MMPs in the healing process of gastric ulcer. Materials and Methods Acetic acid ulcers were produced in male Wistar rats. Gastric tissues were sampled rats on days I, 3, 5, 7, 14, and 28 after the days of induction. Time-dependent changes of Ets-I and MMPs in ulcer margin were examined by Western blotting and RT-PCR. Distribution of Ets-l , MMP-I, MMP-9, and MMP-lO were evaluated by immunohistochemistry and in situ hybridization (ISH). Effects of antisense Ets-I oligonucleotide (50 JLM) on the expressionof Ets-I and MMPs in ulcer healingwere evaluated. ResultsEts-I did not express in the normal gastric mucosa. In the marginal granulationtissues, the nuclei of mesenchymalcells (fibroblast, endothelial cell) and epithelial cells (regenerative cell, damaged mucosal cell) were immunopositive for Ets-I. Ets-I expression was significantly increased at the early phase, and returned to normal levels at the scarred phase. Serial sectioning revealed that fibroblasts and endothelial cells also expressed MMP-l. Expressionsof MMP-9 and MMP-1O were detected in regenerative and damaged epithelium by immunohistochemistry and ISH. Timedependent changes of MMPs were similar to Ets-I. Antisenseoligonucleotide retarded ulcer healing and suppressed the expressions of Ets-I and MMPs. Conclusion These findings suggest that Ets-I plays a key role of angiogenesis and re-epithelization by regulation of MMPs in the early phase of ulcer healing.